Asian Pacific Journal of Tropical Medicine

SELECTED PROCEEDINGS OF THE SECOND BELT AND ROAD TROPICAL MEDICAL ALLIANCE (BRTMA) FORUM IN TROPICAL MEDICINE
Year
: 2019  |  Volume : 12  |  Issue : 13  |  Page : 1--44

Selected proceedings of the Second Belt and Road Tropical Medical Alliance (BRTMA) forum in tropical medicine


 

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. Selected proceedings of the Second Belt and Road Tropical Medical Alliance (BRTMA) forum in tropical medicine.Asian Pac J Trop Med 2019;12:1-44


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. Selected proceedings of the Second Belt and Road Tropical Medical Alliance (BRTMA) forum in tropical medicine. Asian Pac J Trop Med [serial online] 2019 [cited 2019 Oct 15 ];12:1-44
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 Comparison of efficacy of two surgical resection methods for hilar cholangiocarcinoma: A systematic review



Xiao-zhen Li1, Wen-ting Cao1, Wan-ting Xia1, Hai-rong Huang1, Tai-cheng Wang2⊠

⊠Corresponding author: Tai-cheng Wang, Department of Hepatobiliary Surgery, the Second Affiliated Hospital of Hainan Medical University, Haikou 570100, China.

E-mail: wangtaicheng2008@163.com

1School of Public health, Hainan Medical University, Haikou 571199, China

2Department of hepatobiliary surgery, the Second Affiliated Hospital of Hainan Medical University, Haikou 570100, China

Objective: To systematically analyze the prognosis of limited hilar hepatectomy versus combined liver lobectomy in the treatment of hilar cholangiocarcinoma.

Methods: We searched CBM, CNKI, VIP, Wangfang, PubMed, SCI, Elsevier clinicalkey and OVID, andthe cut-off time of searching was 31 April, 2019. Two researchers evaluated the risk of bias of the included studies independently according to the Newcastle-Ottawa Scale (NOS), and extracted data. RevMan 5.3 software was used for meta-analysis.

Results: A total of 12 cohort studies involving 1 288 patients were included, of which 426 patients were in the limited hilar hepatectomy group and 862 patients in the combined liver lobectomy group. Meta-analysis showed that there were statistically significant difference in radical resection rate and postoperative complication rate (RR =0.55, 95%CI 0.40 to 0.74, P<0.001; RR =0.63, 95%CI 0.49 to 0.81, P<0.001) between the hilar limited hepatectomy group and the combined liver lobectomy group; significant differences were found in 1-, 2- and 3-year survival rates (RR =0.75, 95%CI 0.67 to 0.84, P<0.001; RR =0.55, 95%CI 0.42 to 0.74, P<0.001; RR =0.54, 95%CI 0.39 to 0.75, P<0.001) between the two groups.

Conclusions: Comparing with the limited hilar hepatectomy, the radical resection rate of combined liver lobectomy was higher, and the 1-, 2- and 3-year survival rates were improved, but the complications was increased in treatment of hilar cholangiocarcinoma.

Keywords: Hilar Cholangiocarcinoma; Liver resection; Radical rate; Survival rates

Article history:

Received 19 September 2019

Revised 25 September 2019

Accepted 28 September 2019

Available online 7 October 2019

 Analysis of endoscopic screening for upper digestive tract cancer in Danzhou from 2015 to 2017



Xi-wen Liu1, Hua Dong2⊠, Xiao-dan Wang1⊠

⊠Corresponding author: Hua Dong, Cancer prevention center, Hainan cancer hospital, Haikou, Hainan 570100, China. E-mail: 872058575@qq.com

Xiao-dan Wang, School of public health, Hainan Medical University, Haikou, Hainan 571199, China. E-mail: xiaodanwangkk@126.com

1School of Public Health, Hainan Medical University, Haikou, Hainan 571199, China.

2Cancer Prevention Center, Hainan Cancer Hospital, Haikou, Hainan 570100, China

Objective: To analyze and evaluate the results of endoscopic screening for upper digestive cancer in Danzhou city of Hainan, China.

Methods: Totally 300 8 subjects aged from 40 to 69 years old were selected by cluster sampling method for screening of upper digestive tract cancer in Danzhou from 2015 to 2017.

Results: In total, 13 cases (0.43%) of high grade intraepithelial neoplasia and cancer were detected. The detection rate of esophageal cancer, cardia cancer and gastric cancer was 0.07% (2 cases), 0.07% (2 cases) and 0.30% (9 cases), respectively. The diagnosis rate of early cancer was 92.31% (12 cases) and the treatment rate was 46.15% (6 cases).

Conclusion: Endoscopic Screening is an effective way to detect precancerous lesions and early cancer. The detection rate in Danzhou is lower than the national average. Improving the professional and management ability of screening project team, mastering the incidence and mortality trend of upper digestive tract cancer, is key to the implementation and evaluation of the project of early diagnosis and treatment of upper digestive tract cancer.

Keywords: Upper digestive tract cancer; Endoscopic screening; Esophagus neoplasms; Cardia cancer; Gastric neoplasms; Danzhou

Article history:

Received 17 September 2019

Revised 25 September 2019

Accepted 28 September 2019

Available online 7 October 2019

 A new double MEK/PDK1 inhibitor 9za retards cell cycle progression at G0/G1 phase and induces mitochondrial apoptosis in non-small cell lung cancer cells



Rang-ru Liu1,2,3, Zu-tao Yu4, Zhuo Chen4, Dan-qi Liu5, Feng-ying Huang3, Qian-bin Li4, Gao-yun Hu4, Xi-nan Yi6, Xi Li1,2, Hong-hao Zhou1,2, Zhao-qian Liu1,2⊠

⊠Corresponding author: Zhao-qian Liu, Department of Clinical Pharmacology, Xiangya Hospital, Central South University; Institute of Clinical Pharmacology, Central South University; Hunan Key Laboratory of Pharmacogenetics, Changsha, Hunan Province, China.

E-mail: liuzhaoqian63@126.com.

Foundation project: This work was supported by Key Research and Development Project of Hainan Province (Grant No. ZDYF2016128, ZDYF2017092); National Natural Science Foundation of China (Grant No. 21102184, 81573508, 81773640 and 81874327); National Key Research and Development Program of China (Grant No. 2016YFC1306900 and 2016YFC0905002) and Strategy-Oriented Special Project of Central South University in China (Grant No. ZLXD2017003).

1Department of Clinical Pharmacology, Xiangya Hospital, Central South University, Changsha, Hunan Province, China

2Hunan Key Laboratory of Pharmacogenetics, National Clinical Research Center of Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, Hunan Province, China

3Key Laboratory of Tropical Diseases and Translational Medicine of the Ministry of Education & Hainan Provincial Key Laboratory of Tropical Medicine, Hainan Medical University, Haikou, Hainan Province, China

4Department of Medicinal Chemistry, Xiangya School of Pharmaceutical Sciences, Central South University, Changsha, Hunan Province, China

5Department of Pharmacy, Xiangya Hospital, Central South University, Changsha, Hunan Province, China

6The United Laboratory for Neurosciences of Hainan Medical University and the Fourth Military Medical University, Haikou, Hainan Province, China

Objective: To identify the molecule mechanism of potential cytotoxicity and proapoptosis of 9za in non-small cell lung cancer (NSCLC) cells.

Methods: MTT assay, cell cycle detection kit, JC-1 staining assay were adopted to detect the cell viability, the cell cycle distribution and the mitochondrial membrane potential, respectively. Cell apoptosis was measured by Annexin V-FITC/propidium iodide apoptosis detection and the morphology was observed under a light microscope and the colorimetric TUNEL assay. Western blot was used to monitor the cell cycle-, apoptosisrelated proteins and some proteins involved in the pathways.

Results: MTT assay showed that 9za significantly weakened the viability of NSCLC cells. Cell cycle analysis demonstrated that the low concentrations of 9za arrested the cell cycle at G0/G1 phase, which was further confirmed by the down-regulated levels of Cyclin D1, cyclin-dependent kinase 4 and cyclin-dependent kinase 6. In addition, Annexin V-FITC/propidium iodide apoptosis analysis, morphological observations and TUNEL assays revealed that the high concentrations of 9za could induce cell apoptosis. Furthermore, the JC-1 staining assay indicated that the mitochondrial membrane potential was reduced after 9za treatment. Western blot also manifested that 9za dramatically decreased the protein levels of the total Bcl-2, Cytochrome C in the mitochondria and BCL2 associated X in the cytoplasm. However, the levels of BCL2 associated X in the mitochondria, Cytochrome C in the cytoplasm, cleaved caspase-9, cleaved caspase-3 and the ratio of cleaved- PARP1/PARP1 showed the opposite trends. Moreover, the dose-dependently decreased phosphorylation levels of PDK1, protein kinase B, MEK and extracellular signal regulated kinase 1/2 following 9za treatment confirmed that 9za was indeed a double MEK/PDK1 inhibitor as we expected. Coadministration with PD0325901 or BX517 strengthened the cytotoxic and pro-apoptotic effect of 9za, verifying that 9za inhibited cell proliferation and induced cell apoptosis through the double MEK/PDK1 signaling pathways in NSCLC cells.

Conclusions: This study demonstrates that 9za could play the effect of cytotoxicity and pro-apoptosis via the dual MEK/PDK1 signaling pathways in NSCLC cells and provides certain experimental foundation for 9za as a new type of drug candidate against non-small cell lung cancer.

Keywords: 9za; Cytotoxicity; Cell cycle arrest; Mitochondrial apoptosis; Dual MEK/PDK1 signaling pathways; Non-small cell lung cancer

Article history:

Received 9 September 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Penicilazaphilone C, a new azaphilone, induces apoptosis by blocking the Notch signalling pathway in gastric cancer



Huan-ge Zhao1#, Ming Wang2#, Ying-ying Lin1, Song-lin Zhou1⊠

#These authors contributed equally to this work.

⊠Corresponding author: Song-lin Zhou, Key Laboratory of Tropical Translational Medicine of the Ministry of Education and Hainan Provincial Key Laboratory of Tropical Medicine, Hainan Medical University, Haikou, Hainan Province, China. Tel.: +86-898-66893335

E-mail: zhousonglin106@163.com

Foundation project: This work was supported by the National Natural Science Foundation of China (Grant No. 81560484), and the Key Research and Development Project of Hainan Province (Grant No. ZDYF2019177).

1Key Laboratory of Tropical Translational Medicine of the Ministry of Education and Hainan Provincial Key Laboratory of Tropical Medicine, Hainan Medical University, Haikou, Hainan Province, China

2Health and Family Planning Commission of Wanzai County of Jiangxi Province, Yichun, Jiangxi Province, China

Objective: To explore the molecular mechanisms of the anticancer activities of penicilazaphilone C against gastric cancer.

Methods: In vitro effects of penicilazaphilone C on cell growth, proliferation, and apoptosis were evaluated by MTT, BrdU, MTS, colony formation assays, Hoechst 33258 staining, and flow cytometry. Related proteins were examined by Western blotting assays. The expression of Notch receptor was analyzed using real-time PCR. In vivo antitumor activities of penicilazaphilone C were observed in nude mice.

Results: Compared to the controls, penicilazaphilone C suppressed cell proliferation and promoted apoptosis in MGC-803 and SGC-7901 cells. The Notch/PTEN/AKT axis was involved in the activating penicilazaphilone C-induced apoptosis. Penicilazaphilone C decreased levels of Notch, NICD, phospho-PTEN and phospho- AKT compared to controls. The penicilazaphilone C-induced inhibition of Notch-related protein expression levels and the resulting apoptosis could be reversed by overexpression of Notch1 or/and Notch2. Moreover, penicilazaphilone C inhibited tumor growth in mice bearing tumours derived from MGC-803 and SGC-7901 cells, respectively.

Conclusions: Penicilazaphilone C can induce the apoptosis by suppressing the activation of the proteolytic cleavage of the Notch receptor and subsequently blocking the PTEN/AKT signaling axis in gastric cancer cells. Thus, penicilazaphilone C is a potential alternative agent for the treatment of gastric cancer.

Keywords: Penicilazaphilone C; Notch signalling pathway; Gastric cancer; Apoptosis; Molecular mechanism

Article history:

Received 8 September 2019

Reevised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Correlation between glycosylated hemoglobin level and blood pressure among 906 diabetic patients in tropical areas



Guo-tian Lin1, Jie Li2, Yue-cai Yu2, Chan-juan Zhao1, Li-min He1, Zhuo Chen1, Fan Zhang1⊠

⊠Corresponding author: Fan Zhang, School of Public Health, Hainan

Medical University, No.3 Xueyuan Road, Haikou, Hainan Province, China.

E-mail:17724886@qq.com

Tel.: 13617547230

Foundation project: This study was funded by the National Natural Science Foundation of China (NSFC) with Grant No: 81860577.

1School of Public Health, Hainan Medical University, No. 3 Xueyuan Road, Haikou, Hainan Province, China

2The Second Affiliated Hospital of Hainan Medical University, No. 48 Baishuitang Road, Haikou, Hainan Province, China

Objective: To observe the correlation between glycosylated hemoglobin level and blood pressure, and to provide clinical evidence to assist the screening and prediction of hypertension among diabetic patients in tropical setting.

Methods: A total of 906 diabetic patients who had previously received physical examination at the Second Affiliated Hospital of Hainan Medical University from January 2009 to December 2018 were selected. Retrospective case-case study method was used. The average glycosylated hemoglobin level (HbA1c) was (5.80±1.27)%, and average blood pressure level was (129±18/79±10) mmHg. Patients were divided into group A: blood glucose well controlled, HbA1c<6.5%, (58.82±12.16) years old (754 cases); and group B: blood glucose poor controlled, HbA1c=6.5%, (60.22±11.82) years old (152 cases). There was no significant difference in age between the two groups (T=-1.302, P =0.186). T-test, χ2-test, spearman correlation analysis and receiver operating characteristic (ROC) curve analysis were used to observe the correlation between glycosylated hemoglobin level and blood pressure, and the prevalence of hypertension.

Results: Group B showed higher systolic blood pressure (134.19±18.62) mmHg vs (127.76±17.21) mmHg, diastolic blood pressure (81.82±10.92) mmHg vs (79.00±9.42) mmHg and blood glucose level (9.10±3.09) mmol/L vs (5.52±0.95) mmol/L than group A (T values were 4.335, 3.226, 14.142, respectively, P values were all <0.001). Spearman correlation analysis showed that HbA1c was positively correlated with systolic and diastolic blood pressure (the correlation coefficients r were 0.154 and 0.121, respectively, P values were all <0.001). The prevalence rates of hypertension were 31.17% (235/754) in group A, and 43.4% (66/152) in group B, and the difference was statistically significant [χ2=8.562, OR=1.695, 95% CI (1.187-2.419), P =0.003]. ROC curve of glycosylated hemoglobin was drawn to screen hypertension and the results showed that AUG was 0.587 [95% CI (0.548-0.626), P< 0.001], the cut-off point, sensitivity and specificity were 6.05%, 33.6% and 78.2%, respectively.

Conclusions: There was positive correlation of glycosylated hemoglobin and blood pressure and it is a novel view of using HbA1c to screen and predict hypertension among diabetic patients in tropic settings. More researches are needed in this field.

Keywords: Glycosylated hemoglobin; Blood pressure; ROC curve

Article history:

Received 7 September 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Distribution and drug resistance of clinically isolated pathogenic bacteria at a tertiary care hospital in Haikou, China



Fang Lian, Hua Pei⊠, Chong Lin, Shao-wen Chen, Ying-xian Su, Wei-bin Chen, Lin Chen

⊠Corresponding author: Hua Pei, Department of Clinical Laboratory, the Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570311, China.

E-mail: phzmh61@aliyun.com

Foundation project: This research was supported by the National Natural Science Foundation of China (No. 81860002).

Department of Clinical Laboratory, the Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570311, China

Objective: To analyze the distribution and drug resistance of clinical isolated pathogens to provide reference for rational use of antimicrobial agents.

Methods: We retrospectively analyzed the distribution and drug resistance data of clinical bacterial isolates from the Second Affiliated Hospital of Hainan Medical University from Jan 2018 to Dec 2018 and their clinical distribution and drug resistance were analyzed.

Results: The results of comprehensive analysis of 7 897 non-repetitive bacteria showed that 5 044 strains (63.87%) were Gram-negative bacteria, 1 818 strains (23.02%) were Gram-positive bacteria, and 1 035 strains (13.11%) were fungi. Gram-negative bacteria mainly were Pseudomonas aeruginosa (15.44%), Escherichia coli (12.79%) and Acinetobacter baumannii (8.66%). Gram-positive bacteria mainly were Staphylococcus aureus (6.43%), Enterococcus faecium (4.89%) and Enterococcus faecalis (4.65%). The main fungi were Candida albicans (6.34%). Most of the collected and tested samples were urine (32.87%), followed by sputum (30.10%) and wound secretion (13.54%). In emergency department, intensive care unit and urology department, the detection rate of pathogens was most frequently found among three departments. Escherichia coli had higher resistance to ampicillin, tetracyclin and piperacillin, but were sensitive to imipenem and meropenem. Pseudomonas aeruginosa were sensitive to amikacin, tobramycin and gentamycin. Acinetobacter baumannii had higher resistance to cefotaxime, cefepime, and they were sensitive to minocycline. Among Staphylococcus aureus, a total of 123 strains of MRSA (24.21%) were more resistant to penicillin, azithromycin and erythromycin, but sensitive to vancomycin, linezolid and taicoplanin.

Conclusions: The problem of drug resistance of clinical isolated pathogens was serious. The hospital should strengthen the dynamic monitoring of drug resistant bacteria. Antibiotics should be used rationally to reduce the spread of drug resistant strains and effective measures must be taken to prevent infections.

Keywords: Pathogen; Clinical distribution; Antibiotics; Drug resistance

Article history:

Received 12 September 2019

Revised 22 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Effect of electroacupuncture on expression of CREB and its ability to bind to synaptic proteins in rats with post-traumatic stress disorder



Mi Li1#, Kai Li1#, Ning Ding2, Yi-qiang Xie1, Kun Niu1, Hong Zhang3⊠

#These authors contributed equally to this work.

⊠Corresponding author: Hong Zhang, Department of Acupuncture-moxibustion and Tuina, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, Sichuan Province, China.

E-mail: 13548134059@163.com

Foundation project: This study was supported by the National Natural Science Foundation of China (Project No. 81574086) and Hainan Natural Science Foundation (Project No. 819QN223).

1School of Traditional Chinese Medicine, Hainan Medical University, Haikou 571199, Hainan Province, China

2Clinical Medical College, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, Sichuan Province, China

3Department of Acupuncture-moxibustion and Tuina, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, Sichuan Province, China

Objective: To observe the effects of electroacupuncture on the expression of CREB and its ability to blind to synaptic key proteins in rats with post-traumatic stress disorder (PTSD), thus to lay the foundation for further study of the mechanism of electroacupuncture treatment of PTSD (i.e., electroacupuncture promotes the interaction between synaptic plasticity and BDNF-TRKB signaling pathway).

Methods: SD male rats were randomly divided into blank group, model group and electroacupuncture treatment group, with 8 rats in each group. The model of PTSD was established by single continuous stress method (this modeling method is dominated by forced drowning, and tropical disasters are also mostly related to drowning). After modeling, the treatment group received electroacupuncture therapy, followed by retaining the needles for 20 min. The treatment was conducted once daily for 21 d. The behavioral changes were evaluated by spontaneous activity and conditional fear reaction. The expression of CREB was detected by immunohistochemistry and Western Blot. The binding ability of CREB to synaptic protein was verified by CHIP.

Results: Compared with the model group, the spontaneous activity distance of the treatment group rats increased (P<0.01) and the percentage of stiffness time decreased (P< 0.01), the expression of CREB protein in the treatment group was significantly increased (P<0.01), the binding ability of CREB to PSD95 protein was increased in treatment group (P<0.05).

Conclusions: Electroacupuncture regulates the interaction mechanism between synaptic plasticity and BDNFTrkB signaling pathway of the rat model of PTSD, most likely by increasing the binding ability of CREB to the key synaptic protein-PSD95.

Keywords: Single continuous stress; Drowning; Electroacupuncture; Post-traumatic stress disorder; CREB; PSD59; Binding ability

Article history:

Received 11 September 2019

Revised 20 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Distribution of hepatitis B virus genotypes in Hainan, China



Xin-jun Chen1, Fei-fei Yin2,3, Gang Lu2,3, Xiu-ji Cui1,3⊠

⊠Corresponding author: Xiu-ji Cui, Laboratory of Pathogenic Biology and Immunology, Hainan Medical University, Haikou, Hainan, China; Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, Hainan, China.

E-mail: cuixj26@163.com

Foundation project: The project is supported by National Nature Science Foundation of China (NSFC) under grant No. 81660335 and The Natural Science Foundation of Hainan Province, China under grant No. 819QN237.

1Laboratory of Pathogenic Biology and Immunology, Hainan Medical University, Haikou, Hainan, China

2Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China; and The University of Hong Kong, Pokfulam, Hong Kong

3Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, Hainan, China

Objective: To analyze the distribution of hepatitis B virus (HBV) genotypes in Hainan island, which is one of the most endemic areas of HBV infection in China.

Methods: A total of 52 patients with chronic HBV infection were enrolled. HBV genotypes were determined by nested polymerase chain reaction using genotype-specific primer pairs.

Results: All patients were positive for nested polymerase chain reaction and the genotype B was detected in all patient serum samples. Moreover, 23 of 52 patients (44.2%) have been detected with genotype B and C coinfection.

Conclusions: HBV genotype B is mostly prevalent in Hainan island. Intriguingly, co-infection of genotype B and C was frequently detected in Hainan island, but it may need to be further verified by HBV full genome sequencing analysis.

Keywords: HBV; Genotype; Hainan island

Article history:

Received 11 September 2019

Revised 21 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Complete genome sequence of a rodent polyomaviruses in Hainan Island, China



Shan-shan Wang1,2,3, Yue Wu1,2,3, Huan-huan Zhou3, Wen-qi Wang3, You Zhang1,2,3, Wei Zhao3, Xiu-ji Cui3, Li-hua Li3, Jing-long Chen3, Lang-yu Rao3, Jiang Du1,2,3, Fei-fei Yin1,2,3⊠, Gang Lu1,2,3⊠

⊠Corresponding author: Gang Lu, Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China; and The University of Hong Kong, Pokfulam, Hong Kong.

E-mail: ganglu2018@126.com

Fei-fei Yin, Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China; and The University of Hong Kong, Pokfulam, Hong Kong.

E-mail: yinfeifeiff@163.com

Foundation project: This work was supported by the Innovation Research Team Project of Hainan Natural Science Foundation (2018CXTD340) and the National Natural Science Foundation of China (No. 81672072, No.81760378 and No.81860367).

1Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China; and The University of Hong Kong, Pokfulam, Hong Kong

2Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, Hainan, China

3Department of Pathogen Biology, Hainan Medical University, Haikou, Hainan, China

Objective: Polyomavirus infection is reported in a wide range of mammalian and avian hosts, including asymptomatic infection, acute systemic disease, and tumor induction.This study aims to obtain and characterize the complete genome of rodent polyomaviruses, PyVs-HMU. The host of virus was a Rattus norvegicus in the residential areas of Hainan Island, China.

Methods: The liver samples of Rattus norvegicus were collected from the residential areas of Hainan Island, and then processed with a viral particle-protected, nucleic acid purification method. The extracted RNA and DNA were amplified by sequence-independent PCR. The amplified viral nucleic acid libraries for the samples of Rattus norvegicus were then sequenced using an Illumina GAII sequencer for a single read of 100 bp in length. The raw sequence reads were then filtered using previously described criteria to obtain valid sequences.

Results: We obtained the complete genome of a novel polyomaviruses, PyVs-HMU. The genomic sequence of PyVs-HMU has been submitted in GenBank under accession number MK372231. The complete genom of PyVs-HMU is 5318 nucleotides (nt) in length with a G+C content of 45.36%. The complete PyV-HMU sequences display the representative genome organization of polyomaviruses. The genome contain antigens of spliced small T (STAg), middle T (MTAg) and large T(LTAg), and a noncoding control region (NCCR) separate the late region of structural VP1, VP2, and VP3 proteins. The STAg, LTAg, VP1, VP2, VP3 and MTAg encoded proteins of 194, 776, 384, 347, 228 and 414 amino acids (aa) respectively. Phylogenetic analyses depend on LTAg amino acid sequence revealed that the PyV-HMU to be a relative lineage beside a cluster comprising RnorPyV1(KR065723).

Conclusions: The discovery of PyV-HMU expands the geographic range of polyomavirus and will provide further insights into the ecology and evolution of PyVs in rodents and humans. The identification of the novel rodent PyVs will provide basic data for the control of emerging zoonotic infectious diseases.

Keywords: Rodents; Polyomavirus; Genomic analysis

Article history:

Received 19 September 2019

Revised 25 September 2019

Accepted 28 September 2019

Available online 7 October 2019

 Screening and bioinformatics analysis of specific microRNAs in testes of rats exposed to cigarette smoke



Jing Zhang1,2#, Li-juan He2#, Jing-chao Ren3#, Yun-fei Huang2, Kai-ju Liao4, Chun-xue Zhong2, Ying Zou2, Guang-hui Zhang3⊠, Chen Zhang2⊠

#These authors contributed equally to this work.

⊠Corresponding author: Guang-hui Zhang, Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Public Health, Xinxiang Medical University, 601 Jinsui Road, Xinxiang, Henan Province, China.

E-mail: zhgh221@163.com.

Chen Zhang, College of Public Health, Xinjiang Medical University, 393 Xinyi Road, Urumqi, Xinjiang Province, China.

E-mail: zhangchenxjmu@163.com.

Foundation project: This work was supported by the Natural Science Foundation of China (No. 81260433 and No. 81660547)

1College of Public Health, Hainan Medical University, No. 3 Xueyuan Road, Haikou, Hainan Province, China

2College of Public Health, Xinjiang Medical University, 393 Xinyi Road, Urumqi, Xinjiang Province, China

3Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Public Health, Xinxiang Medical University, 601 Jinsui Road, Xinxiang, Henan Province, China

4Health Emergency Center, Chinese Center for Disease Control and Prevention, 155 Changbai Road, Beijing, 102206, China

Objective: To identify effects of cigarette smoke on the male reproductive capacity and to explore the miRNA expression in the testes after cigarette smoke exposure.

Methods: Eighty male rats were conducted by factorial analysis of variance designed for cigarette exposure. A microarray was employed to detect the differential expression of miRNA in the testis tissue of smoke-exposed rats.

Results: Four miRNAs (miR-138-5p, miR-181d-5p, miR-19a-3p, and miR-3588) were significantly downregulated and one miRNA (miR-155-5p) was upregulated in the testes of smoke-exposed rats compared with control rats. This result was further confirmed by a quantitative RT-PCR assay, and pathological changes were observed in the testes. Bioinformatics analysis showed that the predicted target genes were closely related to the regulation of the apoptosis pathway.

Conclusions: miRNA may play an important role in the smoke-exposure-induced testicular toxicity of male rats.

Keywords: Cigarette smoke; Rat; Testis; microRNA; Testicular toxicity

Article history:

Received 10 September 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Prevalence and genotyping of Enterocytoon bieneusi in farmed black goats from Hainan Province, the southernmost part of China



Huan-huan Zhou1,2,3, Xin-li Zheng4, Tian-ming Ma1,2,3, Zong-xi Cao4, Zhe Chao4, Li-min Wei4, Quan-wei Liu4, Rui-ping Sun4, Feng Wang4, Yan Zhang4, Wei Zhao1,2,3⊠, Gang Lu1,2,3⊠

⊠Corresponding authors: Gang Lu, Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, 571199, China.

E-mail:luganghn@aliyun.com

Wei Zhao, Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, 571199, China.

E-mail: hayidazhaowei@163.com

Foundation project: This work was supported by the Innovation Research Team Project of Hainan Natural Science Foundation (2018CXTD340); the National Natural Science Foundation of China (No. 81672072 and No. 81760378); the Young Talents Science and Technology Innovation Project of Hainan Association for Science and Technology (QCXM201802); the Hainan major science and technology project (ZDKJ2016017-01).

1Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, China

2Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China

3Key Laboratory of Translation Medicine Tropical Diseases, Hainan Medical University, Haikou, Hainan, 571199, China

4Institute of Animal Science and Veterinary Medicine, Hainan Academy of Agricultural Sciences, Haikou 571100, China

Objective: To investigate the infection rate and genotype distribution of Enterocytoon (E.) bieneusi in farmed black goats from the Hainan Province, China.

Methods: A total of 341 fresh fecal samples were collected from black goats farmed in five different locations of the Hainan Province, China. E. bieneusi was examined and genotyped through PCR and sequencing analysis of the internal transcribed spacer (ITS) region of this pathogen.

Results: The average prevalence of E. bieneusi in black goats from from the five locations was 24.0% (82/341) ranging from 6.3% (4/63) to 37.2% (32/86) (χ2=17.252, P<0.01). The detected 82 E. bieneusi isolates belonged to eight ITS genotypes including six known genotypes (AHG1, CHG2, CHG3, CHG5, CM21 and D) and two novel genotypes (HNG-I and HNG-II). Amongst the genotypes, CHG5 was the the most prevalent with a prevalence of 57.3% (47/82), followed by CHG3 (28.0%, 23/82), CHG2 (4.9%, 4/82), CM21 (3.7%, 3/82), D (2.4%, 2/82), AHG1 (1.2%, 1/82), HNG-II(1.2%, 1/82) and HNG-II(1.2%, 1/82). In those genotypes, only genotype D was found in humans previously.

Conclusions: This represents the first report identifying E. bieneusi in black goats from Hainan Province of China. The results indicate that E. bieneusi has a high prevalence and a wide distribution in those animals from Hainan Province, but the risk of zoonotic transmission of E. bieneusi from them to human is low.

Keywords: Enterocytozoon bieneusi; Genotype; ITS region; Goats; Hainan

Article history:

Received 5 September 2019

Revised 25 September 2019

Accepted 26 September 2019

Available online 7 October 2019

 Occurrence of Cryptosporidium, Enterocytozoon bieneusi and Blastocystis genotypes in Rousettus leschenaultii collected from an urban public park of Hainan Province, China



Wei Zhao1,2,3,4, Huan-huan Zhou1,2,3, Tian-ming Ma1,2,3, Gang Lu1,2,3⊠

⊠Corresponding author: Gang Lu, Department of Pathogenic Biology, Hainan Medical University, Xueyuan Road 3, Haikou, Hainan, China.

E-mail: luganghn@163.com; ganglu2018@163.com

Foundation project: This work was supported by the Innovation Research Team Project of Hainan Natural Science Foundation (2018CXTD340); the National Natural Science Foundation of China (No. 81672072 and No.81760378); the Open Subject of Key Laboratory of Parasite and Vector Biology, MOPH (WSBKFKT-201802).

1Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, China

2Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China

3Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou 571199, China

4Department of Parasitology, Wenzhou Medical University, Wenzhou, Zhejiang Province, 325035, China

Objective: To obtain information about the occurrence of Blastocystis spp., Cryptosporidium spp., and Enterocytozoon (E.) bieneusi in fruit bats (Rousettus leschenaultii) collected from an urban public park Hainan Province, China and to analyze the genetic characteristics of the obtained parasites carried by those bats.

Methods: On 4th June 2019, ten piles of fresh faecal sample of fruit bats were collected from the Wanlvyuan Gardens in central Haikou, Hainan of China. Blastocystis spp., Cryptosporidium spp., and E. bieneusi were examined by sequencing analysis of the small subunit rRNA gene or internal transcribed spacer (ITS) gene.

Results: Among the 10 DNA specimens analyzed, seven (70.0%) were positive for Cryptosporidium spp. and two (20.0%) were positive for E. bieneusi but none of them were positive for Blastocystis. For Cryptosporidium, two novel genotypes were identified which shared 98.2% and 94.4% homology with Cryptosporidium (C.) andersoni Type C, respectively, and were named as C. andersoni Type D (in 6 specimens) and E (in one specimens). The two E. bieneusi-positive isolates were identified as a known zoonotic genotype (PigEbITS7) and a novel genotype (named HNB-I) respectively.

Conclusions: The finding of C. andersoni and E. bieneusi genotype PigEbITS7 in fruit bats in Hainan, China suggests that these parasites carried by fruit bats can be transmitted to other animals and humans to cause zoonotic infections.

Keywords: Cryptosporidium; Enterocytozoon bieneusi; Blastocystis; genetic characterization; bats

Article history:

Received 20 September 2019

Revised 22 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Treating nonspecific low back pain by SET sling exercise therapy combined with tropical herb-Rattan Moxibustion liquid external rubbing therapy



Kun Niu#, Qian Niu#, Yi-qiang Xie, Fan Yang⊠

#These authors contributed equally to this work.

⊠Corresponding author: Fan Yang, College of Traditional Chinese Medicine, Hainan Medical University, Haikou 571199, China.

E-mail:icelanyang@163.com

Foundation project: This study was funded by the Natural Science Foundation of Hainan Province (Grant No. 818QN248), the National Natural Science Foundation of China (Grant No. 81574095), Educational Science Research Foundation of Hainan Medical University (Grant No. HYP201714).

College of traditional Chinese Medicine, Hainan Medical University, Haikou 571199, China

Objective: To explore the clinical effects of tropical Eucommia ulmoides compound-Rattan Moxibustion liquid external rubbing therapy which is from medicine of Li Nationality in Hainan combined with SET sling exercise therapy technique and lumbar-abdominal muscle training program on treating nonspecific low back pain (NLBP) as well as its influence on serum 5-hydroxytryptamine and beta-endorphin levels.

Methods: From June 2016 to June 2018, 42 NLBP patients were admitted to the outpatient and ward of Rehabilitation Department of the First Affiliated Hospital of Hainan Medical College, who were randomly divided into control group (n=21) and experimental group (n=21). On the basis of traditional massage manipulation, the control group adopts lumbar and abdominal muscles training, while Rattan Moxibustion liquid external rubbing therapy combined with SET sling exercise therapy was used in the experimental group. The treatment was given once every other day, three times a week and 20 min each time for 8 weeks. Then ODI index and VAS scale of low back pain, as well as serum 5-HT and beta-EP levels were observed and detected.

Results: The improvement in low back pain index ODI index of the experimental group were better than that of the control group (control group from 21.23±1.57 to 4.78±0.86 while experiment group from 21.27±1.36 to 2.75±0.64 after 8 weeks' treatment)(P<0.05). VAS scale decreased significantly in the experiment group compared with control group (control group from 6.62±1.51 to 2.71±0.92 while experiment group from 6.71±1.55 to 1.83±0.79 after 8 weeks' treatment)(P<0.05). Serum 5-HT in the experimental group were significantly improved compared with those in the control group [control group from (70.15±20.64) ng/mL to (127.34±40.59) ng/mL while experiment group from (68.72±19.18) ng/mL to (142.65±38.72) ng/mL after treatment](P<0.05) and beta-EP level ameliorates compared with control group [control group from (180.63±51.87) pg/mL to (225.76±57.63) pg/mL while experiment group from (172.39±50.72) pg/mL to (251.26±62.89) pg/mL after treatment](P<0.05).

Conclusions: Tropical herb-Rattan Moxibustion liquid external rubbing therapy combined with SET sling exercise therapy technology shows significant effects on NLBP. The mechanism may be related to the improved serum 5-HT and beta-EP levels.

Keywords: Nonspecific low back pain; SET sling exercise therapy; Tropical Herb-Rattan moxibustion liquid external rubbing therapy; Eucommia ulmoides compound

Article history:

Received 4 September 2019

Revised 20 September 2019

Accepted 26 September 2019

Available online 7 October 2019

 Alpinia oxyphylla alleviates diabetic nephropathy through the NLRP3 inflammasome pathway in rats



Mi Li#, Kai Li#, De-hui Yin, Ye Zhu, Yi-qiang Xie⊠

#These authors contributed equally to this work.

⊠Corresponding author: Yi-qiang Xie, College of Traditional Chinese Medicine, Hainan Medical University, Haikou 571199, China.

E-mail: 13036001921@163.com

Foundation project: This study was funded by the Natural Science Foundation of Hainan Province (Grant No. 2019CXTD407, 819QN228), the National Natural Science Foundation of China (Grant No. 81860836) ,Scientific research Foundation of Hainan Medical University (Grant No. HYPY201924).

College of Traditional Chinese Medicine, Hainan Medical University, Haikou 571199, China

Objective: Previous studies have confirmed that the tropical traditional Chinese herb-Alpinia (A.) oxyphylla are effective on diabetic nephropathy. The purpose of this experiment was to investigate whether A. oxyphylla could prevent and treat diabetic nephropathy by regulating NLRP3 inflammasome pathway.

Methods: Male SD rats and NLRP3 gene silencing rats were assinged into blank group, model group, A. oxyphylla group, valsartan group and NLRP3 gene silencing group, with 8 rats in each group. The model was established by intraabdominal injection of streptomycin after feeding with high sugar and high fat diet for 6 weeks. Based on successful modeling, administration protocal for a period of 8 weeks were performed. A. oxyphylla group was given A. oxyphylla decoction (2.6g/kg/d ), while the model group and NLRP3 gene silencing group were given saline as the same dosage of A. oxyphylla decoction. The valsartan (10 mg/kg/d) was given to the valsartan group. After 8 weeks of treatments, blood glucose, 24 h urinary protein, creatinine, urea nitrogen levels were detected, and the pathological sections of the kidney were examined. The expression of NLRP3 protein was detected by Western-Blot and PCR. NLRP3 protein, caspase-1, IL-18 and IL-1β in renal tubules were detected in each group by immunofluorescence and immunohistochemistry.

Results: Compared with the model group, A. oxyphylla significantly reduced blood glucose, 24 h urinary protein, creatinine, urea nitrogen levels (P all<0.05) but the pathological injury in kidney was less. The expression of NLRP3 protein was negative in NLRP3-/-group while positive in the model group, A. oxyphylla group and valsartan group. Expression of NLRP3 protein, NLRP3, caspase-1, IL-18 and IL-1β were also significantly lower in A. oxyphylla trated group compared with the model group (P all<0.05).

Conclusions: A. oxyphylla could inhibit the expression of NLRP3 protein, reduce IL-18 and IL-1βand other inflammatory factors and alleviate the damage of kidney caused by inflammatory reaction. This is one of the mechanisms whereby A. oxyphylla prevents and treats diabetic nephropathy

Keywords: Alpinia oxyphylla; Diabetic nephropathy; Inflammation; NLRP3

Article history:

Received 4 September 2019

Revised 13 September 2019

Accepted 15 September 2019

Available online 7 October 2019

 Adsorption kinetics and thermodynamics of phenylethanoid glycosides on macroporous resins in the leaves of Callicarpa nudiflora



Yong Wang1,2⊠, Ming-ming Zou1

⊠Corresponding author: Yong Wang, School of Pharmacy, Hainan Medical University, Haikou 571199, China; Hainan Provincial Key Lab of R&D of Tropical Herbs, Haikou 571199, China

E-mail: wangyong1982_2004@aliyun.com

1School of Pharmacy, Hainan Medical University, Haikou 571199, China

2Hainan Provincial Key Lab of R&D of Tropical Herbs, Haikou 571199, China

Objective: To investigate the adsorption kinetics and thermodynamic characteristics of phenylethanoid glycosides (PGs) in the leaves of Callicarpa nudiflora on macroporous resins and provide a reference for the separation and purification of these compounds.

Methods: With adsorption and desorption ratio as indexes, optimum types of macroporous resin for purification of PGs were selected from 8 kinds of macroporous resins by static adsorption and desorption tests, and then adsorption kinetics model and adsorption isotherm model of PGs were established to investigate their adsorption processes.

Results: SP-825 and SP-207 resin were selected and they have similar adsorption process for PGs. Both of them showed a fast adsorption in 0-60 min, a slow adsorption in 60-360 min, and an equilibrium adsorption stage after 360 min. Adsorption dynamic behavior was well described by quasi-second-order equation of both SP-825 and SP-207 macroporous resins, and adsorption rate was mainly controlled by liquid film diffusion and intraparticle diffusion. Equilibrium adsorption data fitted Langmuir and Freundlich isotherm equations well. Both of the two kinds of resins showed good adsorption properties for PGs, and the adsorption process belongs to favorable adsorption.

Conclusions: Both of the kinetic model and thermodynamic model can well describe the adsorption process of SP-825 and SP-207 macroporous resins and the two resins were regarded as excellent adsorption resins for the purification of PGs from the leaves of Callicarpa nudiflora.

Keywords: Callicarpa nudiflora; Phenylethanoid glycosides; Macroporous adsorption resin; Adsorption kinetics; Adsorption thermodynamics

Article history:

Received 18 September 2019

Revised 24 September 2019

Accepted 28 September 2019

Available online 7 October 2019

 Neurovascular mechanism whereby tropical-herb Rattan Moxibustion treating diabetic peripheral neuropathy of blood stasis type



Qian Niu#, Fan Yang#, Yi-qiang Xie, Kun Niu⊠

#These authors contributed equally to this work.

⊠Corresponding author: Kun Niu, College of Traditional Chinese Medicine, Hainan Medical University, Haikou 571199, China.

E-mail: kunniuniu@163.com

Foundation project: This study was funded by the Natural Science Foundation of Hainan Province (Grant No. 818MS061, 818QN248), the National Natural Science Foundation of China (Grant No. 81860836).

College of Traditional Chinese Medicine, Hainan Medical University, Haikou 571199, China

Objective: To observe the neurovascular mechanism of Rattan Moxibustion in the treatment of blood stasis type diabetic peripheral neuropathy.

Methods: Seventy db/db mice were randomly divided into 7 groups including normal group, blood stasis model group, α-lipoic acid group, fake Rattan Moxibustion group, and three subgroups of Rattan Moxibustion-15-min, 20-min, and 25-min group respectively. After 6 weeks' Rattan Moxibustion therapy, thermal pain threshold, blood glucose, endothelin, nitric oxide, nitric oxide synthase and malondialdehyde levels were detected, and pancreatic tissues morphology were observed under light microscopy. Heme oxygenase 1 (HO-1) proteins in dorsal root neuron were detected by Western blotting.

Results: Compared with normal group, blood stasis model group, fake Rattan Moxibustion group, thermal pain threshold of Rattan Moxibustion group was significantly improved (P<0.05); the blood glucose, endothelin, nitric oxide, nitric oxide synthase and malondialdehyde levels of the 20-min group were significantly ameliorated (P<0.01). In the Rattan Moxibustion groups, the chronic inflammation was found to be reduced according to hematoxylin-eosin staining results and Rattan Moxibustion groups HO-1 expression has been strengthened.

Conclusions: Rattan Moxibustion therapy can significantly improve peripheral nerve function in diabetic peripheral neuropathy of blood stasis type in mice, improve the blood stasis of pathological state; it can also reduce the chronic inflammation of the pancreas. The mechanism may ameliorate vasoactive and antioxidant factors, enhance HO-1 expression, ultimately improve microcirculation and enhance antioxidant capacity of the model mice.

Keywords: Tropical herb; Rattan Moxibustion; Diabetic peripheral neuropathy; Neurovascular

Article history:

Received 11 September 2019

Revised 20 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Radix Morinda officinalis and Cortex cinnamomi treat depression by upregulating 5-hydroxytryptamine, norepinephrine, and dopamine in hippocampus in rats



Yang-zhi Peng#, Yue Su#, Yong Jiang⊠

#These authors contributed equally to this paper.

⊠Corresponding author: Dr. Yong Jiang, Basic Medical College, Chengdu University of Traditional Chinese Medicine, Twelve bridges 37, Chengdu, Sichuan province, China.

E-mail: jiangyong@cdutcm.edu.cn

Basic Medical College, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China

Background: Chinese herbal medicine have been reported to have less side effects in treatment of depression disorder than Western antidepressants, while the mechanism remains unclear. Our previous studies have shown that combined use of Radix Morinda officinalis and Cortex cinnamomi have antidepressant effects

Objective: To explore the mechanism of combined use of Radix Morinda officinalis and Cortex cinnamomi in treating depression disorder.

Methods: Sixty SD rats were randomly divided into experimental group, control group, model group and blank group with 15 rats in each group. After establiment of depression models, the experimental group and control group were given the Chinese decoction (2 mL/day) and fluoxetine hydrochloride (2 mL/day) respectively for 3 weeks, meanwhile the model group and blank group were fed with normal saline (2 mL/day). Body weight measurement and sucrose preference test were performed regularly. Finally, the rats were sacrificed after treatment, and the hippocampus were taken to detect 5-hydroxytryptamine, norepinephrine, and dopamine contents.

Results: The experimental group showed increased body weight and sucrose consumption than the other groups. Higher 5-hydroxytryptamine, norepinephrine and dopamine contents were also observed in the experimental group than other groups.

Conclusions: The antidepressant effects of Radix Morinda officinalis and Cortex cinnamomi decoction may show antiexpression effects by up-regulating content of 5-HT, NE, and DA in rats' hippocampus.

Keywords: Radix Morinda officinalis; Cortex cinnamomi; Depression disorder; Mechanism

Article history:

Received 19 September 2019

Revised 21 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Protective effect of tropical herb-Alpinia oxyphylla compound formula on diabetic nephropathy mice



Fan Yang#, Qian Niu#, Yi-qiang Xie, Kun Niu⊠

#These authors contributed equally to this work.

⊠Corresponding author: Kun Niu, College of Traditional Chinese Medicine, Hainan Medical University, Haikou 571199, China.

E-mail: kunniuniu@163.com

Foundation project: This study was funded by the Natural Science Foundation of Hainan Province (Grant No. 819QN232, 2019CXTD407), the National Natural Science Foundation of China (Grant No. 81860836), Educational Science Research Foundation of Hainan Medical University (Grant No. HYP201814).

College of Traditional Chinese Medicine, Hainan Medical University, Haikou 571199, China

Objective: To observe the mechanism of Alpinia (A.) oxyphylla in the treatment of diabetic nephropathy by regulating blood glucose level.

Methods: A total of 40 db/db diabetic nephropathy mice were randomly divided into four groups including model group, irbesartan group, A. oxyphylla compound formula high dose group and A. oxyphylla compound formula low dose group with 10 in each group. Ten normal mice served as normal control group. Normal group and model group were given normal saline only. Irbesartan group and A. oxyphylla compound formula high and low dose group were given corresponding drugs once a day for 4 weeks (about 0.2 mL, once a day). Blood glucose, BUN, urine protein, SCr excretion, GSH and CAT activity were detected.

Results: Compared with normal group, the blood glucose of model group were increased significantly (P<0.05). While compared with model group, the blood glucose of irbesartan group and A. oxyphylla compound formula groups (both high and low dose) decreased significantly after 4 weeks' treatment. Compared with model group, 24 h urine protein, BUN and SCr of A. oxyphylla compound formula groups (both high and low dose) decreased significantly(P<0.05) after 4 weeks' treatment. Compared with normal group, GSH, CAT of model group decreased significantly (P<0.05). While compared with model group, GSH, CAT of irbesartan group and A. oxyphylla compound formula high dose group increased significantly (P<0.05).

Conclusions: A. oxyphylla compound formula can protect diabetic nephropathy mice by reducing the blood glucose level, decreasing the excretion of urine protein, BUN, SCr, and increasing the activites of CAT and GSH.

Keywords: Mechanism; Tropical herb; Alpinia oxyphylla compound formula; Diabetic nephropathy; Blood glucose level

Article history:

Received 11 September 2019

Revised 20 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Status of HIV infection and its influencing factors in the MSM population of Haikou city, China, in 2018



Qiu-juan Lu1, Bing Zhang2, Li Cao2⊠

⊠Corresponding author: Li Cao, School of Public Health, Hainan Medical University, Haikou, Hainan, 571199, China.

E-mail: 1037822186@qq.com

Foundation project: This study is supported by Hainan Natural Science Foundation (No. 117148).

1Haikou Center for Disease Control and Prevention, Haikou, Hainan 571199, China

2School of Public Health, Hainan Medical University, Haikou, Hainan 571199, China

Objective: To understand the current status of AIDS epidemic among men who had sex with men (MSM) in Haikou, and to analyze the influencing factors to assess the risk of HIV/AIDS infection in MSM group, and then provide a theoretical basis for MSM population to prevent HIV transmission.

Methods: Snowball sampling method was used to find men who had homosexual intercourse in oral sex oanal sex in the past year and conducted a questionnaire survey of them. Blood samples were collected to detect three antibodies against: HIV, syphilis and hepatitis C (HCV). Excel 2007 and SPSS22.0 software were used for statistical description, χ2 test and logistic regression analysis.

Results: A total of 408 MSMs were investigated in this study, 27 HIV infections were detected, with a positive rate of 6.6%. Univariate analysis showing there were statistically significant differences in the prevalence of AIDS in ethnic minorities, local residence time, condom use for same-sex anal sex in the past 6 months, people with sexually transmitted diseases in recent one year, syphilis, and gonorrhea (P<0.05). Logistic regression analysis showed that minority nationality and HIV infection sexually transmitted disease in the past year were risk factors for HIV infection, and condom use for same-sex anal sex in the past 6 months, the opposite sex occurred in the last 6 months and high education level are protective factors for HIV infection.

Conclusion: The HIV infection rate of the MSM population in Haikou is high. It is necessary to increase the use of condoms in the MSM population. For low-education MSM, it is necessary to strengthen the dissemination and education of AIDS-related knowledge, so as to reduce the spread of AIDS in the MSM population.

Keywords: Men who had sex with men; HIV; Status; Influencing factor

Article history:

Received 17 September 2019

Received in revised form 26 September 2019

Accepted 30 September 2019

Available online 7 October 2019

 Prevalence and future prospects of HIV in Pakistan



Muhammad Sohail Afzal1⊠, Warda Hussain2, Muhammad Mateen1, Muhammad Khurram1, Muhammad Zaid1, Muhammad Ali1, Haroon Ahmed3

⊠Corresponding author: Muhammad Sohail Afzal, Department of Life Sciences, School of Science, University of Management and Technology (UMT), Lahore, Pakistan.

E-mail: sohail.ncvi@gmail.com

1Department of Life Sciences, School of Science, University of Management and Technology (UMT), Lahore, Pakistan

2Department of Pathology, Nawaz Sharif Medical College, University of Gujrat, Gujrat.

3Department of Biosciences, COMSATS University Islamabad, Pakistan

In Pakistan, 1.5 million people are living with HIV and only 8% of them are receiving antiretroviral therapy. Moreover, the number of newly reported cases of HIV have increased to 30% and it has caused 78% deaths of the HIV infected patients since 2010. More than eight outbreaks have been reported in different area of Pakistan in past two decades, Gujrat-a city of Punjab is one of them. Recently, a medical practitioner in the civil hospital of Gujrat referred 905 cases which were suspected on the basis of symptoms-rapid weight loss, extreme tiredness, prolonged swelling of lymph glands, mouth sores. HIV-ELISA test was performed for suspected cases and 17.5% cases were reported HIV positive. Female percentage (58%) was higher as compared to the male (42%) with HIV positive cases. Study participants were categorized into different age groups and the HIV was highly prevalent in the individuals with 30-39 years of age (38%), 23% in age group III with (20-29 years). However, 4% cases were less than 10 years of age. Overall, 17% HIV positive cases in a small town of Gujrat are an eye opener to the incidence and prevalence of HIV. Stringent actions are required in collaboration with federal and provincial agencies to control the HIV spread.

Keywords: HIV; Epidemiology; Prevention; Community education; Pakistan

Article history:

Received 4 September 2019

Revised 13 September 2019

Accepted 15 September 2019

Available online 7 October 2019

 An update on HIV in transgenders of Pakistan



Aisha Khan1, Qudsia Rasib1, Sajida Riaz1, Isthiaq Ali2, Naveeda Riaz3, Muhammad Sohail Afzal4, Muhammad Asif Gondal1, Haroon Ahmed1⊠

⊠Corresponding author: Haroon Ahmed, Department of Life Sciences, University of Management & Technology (UMT), Lahore, Pakistan.

E-mail: haroonahmed@comsats.edu.pk; haroonahmad12@yahoo.com

1Department of Biosciences, COMSATS University Islamabad (CUI), Islamabad, Pakistan

2Department of Microbiology, Quaid-i-Azam University, Islamabad, Pakistan

3Department of Biotechnology, International Islamic University (IIUI), Islamabad, Pakistan

4Department of Life Sciences, University of Management & Technology (UMT), Lahore, Pakistan

Objective: HIV is a worldwide contagious disease and recent emergence in transgenders reflecting the alarming situation in Pakistan. Transgender people are one of the most affected groups by the HIV epidemic and are 49 times more likely to be living with HIV than the general population. HIV is an emerging disease in Pakistan. Transgender people in Pakistan often face legal, economic, social and religious exclusion. Even though research on HIV/AIDS has conducted since several years ago, still we have a little knowledge about HIV in transgender people. This study aims to investigate the current status of HIV infection among Pakistan transgender population.

Methods: Overall cases of HIV in 2017 were retrieved from all four provinces of Pakistan by using all available published data. Results: In 2017, totally 66 264 positive HIV cases were reported in transgenders from Punjab (n=52 656), Sindh (n=13 596) and KPK (n=22) provinces of Pakistan. In Sindh province majority of the patients were from Karachi (n=9 123) followed by Hyderabad (n=1 062), Sukkur (n=1 609), Larkana (n=1 130), Mirpurkhas (n=289) and Nawabshah (n=383), respectively while in Khyber Pakhtunkhwa (KPK), the number of HIV/AIDS transgender patients increased up to 450, out of which 22 cases were registered under provincial AIDS control program in 2017. Conclusions: More efforts by international agencies are needed to combat HIV among transgender communities in Pakistan. Government and NGOs should collaborate to find some strategies for control and prevention of HIV. Social and human rights activists should help and encourage them mainly at the point where their legal rights are not respected. Transgender communities and health department should go hand in hand. Initiatives should be taken to link social justice, human rights, health and advocacy to prevent HIV among transgender in near future.

Keywords: HIV; Update; Transgenders; Pakistan

Article history:

Received 19 September 2019

Received in revised form 25 September 2019

Accepted 28 September 2019

Available online 7 October 2019

 Epidemiological characteristics of malaria from 1951 to 2018 in Sanya city of Hainan Province, China



Wen-ting Cao1, De-lei Wu2, Jing Zhou1, Mei-xiang Li1, Shao-ping Wang2⊠

⊠Corresponding author: Shao-ping Wang, Parasitology Department, the Center for Disease Control and Prevention of Sanya, Sanya, Hainan 572000, China.

E-mail: wangshaoping1273@163.com

1Department of Medical Statistics & Epidemiology, School of Public Health, Hainan Medical University, Haikou, Hainan 571199, China

2Parasitology Department, the Center for Disease Control and Prevention of Sanya, Sanya, Hainan 572000, China

Objective: To analyze the progress of eliminating malaria and the epidemiological characteristics of malaria in Sanya city, Hainan Province, China, from 1951 to 2018.

Methods: This study used the historical data analysis method which belongs to descriptive study to analyze the epidemiological characteristics of malaria. The data were obtained from previous historical records of the Center for Disease Control and Prevention in Sanya. Microsoft Excel 2010 and IBM SPSS 21.0 software were used for data collation and analysis.

Results: The malaria prevention and control in Sanya experienced three main stages. Time during 1951-1983 is the period of controlling malaria epidemic, 1984-2009 is the period of regular management, and 2010-2018 is the period of eliminating malaria. From 1951 to 2018, the incidence of malaria showed a long-term trend of gradual decline. The average annual incidence rates were 268.55 per 10 000 people during 1951-1983, 22.96 per 10 000 people during 1984-2009, and 0.026 per 10 000 people during 2010-2018, respectively. The highest incidence of malaria in history was in 1956 and the incidence was 1 030.60 per 10 000 people. From 1951 to 2018, the incidence of malaria was high in all months througout the year, with the highest incidence in July. The distribution characteristics of the insect species were as follows: before 1981, the reported cases were mainly Plasmodium falciparum malaria; from 1981 to 2014, the reported cases were Plasmodium falciparum malaria and Plasmodium vivax malaria; and the local cases were quartan malaria in 2015. As for the characteristics of the vectors: before 1960, Anopheles minimus was the main vector of malaria transmission, and after 1980, Anopheles dirus was the main vector.

Conclusions: From 1951 to 2018, the incidence of malaria in Sanya decreased gradually and showed a trend of elimination.

Keywords: Malaria; Incidence rate; Epidemiological distribution characteristics; Retrospective analysis

Article history:

Received 12 September 2019

Revised 21 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Emerging trend of leishmaniosis in Pakistan: Current status and future prospects



Aisha Khan1, Muhammad Sohail Afzal2, Muhammad Isthiaq Ali3, Naveeda Riaz4, Haroon Ahmed1⊠

⊠Corresponding author: Dr. Haroon Ahmed, Assistant Professor, Infectious Diseases Division, Department of Biosciences, COMSATS University Islamabad (CUI), Park Road, Chakh shahzad, Islamabad, Pakistan.

E-mail: haroonahmad12@yahoo.com; haroonahmed@comsats.edu.pk

1Department of Biosciences, COMSATS University Islamabad (CUI), Islamabad, Pakistan

2Department of Life Sciences, University of Management & Technology (UMT), Lahore, Pakistan

3Department of Microbiology, Quaid-i-Azam University, Islamabad, Pakistan

4Department of Biotechnology, International Islamic University (IIUI), Islamabad, Pakistan

Leishmaniosis is a disease caused by a Leishmania protozoan parasite, which is transmitted by the bite of an infected sand fly. Leishmaniasis is among the most dangerous and neglected tropical diseases. In Pakistan, limited research work has been done yet on this neglected disease. Overall cases of leishmaniasis were retrieved from Khyber Pakhtunkhwa (KPK) and Baluchistan provinces from 2018 to 2019. The major risk factors responsible for these outbreaks were also determined. Recent outbreak resulted in more than 28 000 cases of leishmaniasis in KPK from November 2018 to April 2019. Many of the cases were children and approximately 7 810 diagnosed patients did not receive treatment with meglumine antimoniate. Leishmaniasis cases were also identified in the tribal areas of Khyber (n=9 378), Mohmand (n=5 373), Kurram (n=525), Orakzai (n=163), and Karak (n=554). In Balochistan, around (n=5 000) cases of leishmaniasis were reported in 2018 from the District of Killa Abdulla. The poor infrastructure and sanitary conditions increase the breeding and resting sites of sandflies. The local population's poor socio-economic conditions increase access to sandflies and the risk of acquiring leishmaniasis. In Pakistan, leishmaniasis is currently neglected by the health authorities, largely due to a lack of data on the number of cases and the monetary and non-monetary impacts that these cases are having on the country. There is a dire need for outreach activities to provide information to communities about the disease and preventive measures. This is a call to the national and international research communities, and to the WHO, to take notice of the threat of leishmaniasis in Pakistan and to help devise a strategy to address this neglected yet important public health problem.

Keywords: Leishmaniasis; Status; Outbreak; Pakistan

Article history:

Received 4 September 2019

Revised 13 September 2019

Accepted 15 September 2019

Available online 7 October 2019

 A century of onchocerciasis in Sierra Leone



Ibrahim Kargbo-Labour1⊠, Yakuba M. Bah1, Abdulai Conteh1, Mohamed S. Bah2, Jusufu Paye2, Mustapha Sonnie2, Amy Veinoglou3, Joseph B. Koroma4, Mary Hodges2, Yao-bi Zhang5

⊠Corresponding author: Ibrahim Kargbo-Labour, Neglected Tropical Disease Program, Ministry of Health and Sanitation, Sierra Leone.

E-mail: kargbolabour@gmail.com

1Neglected Tropical Disease Program, Ministry of Health and Sanitation, Sierra Leone

2Helen Keller International, Freetown, Sierra Leone

3Helen Keller International, Headquarters New York, USA

4Family Health International 360, Accra, Ghana

5Helen Keller International, Regional Office for Africa, Dakar, Senegal

In 1926, Blacklock first described onchocerciasis (OV) transmission by black fly, Simulium damnosum in Sierra Leone. In the 1950-1960's, high OV prevalence was demonstrated along the rivers and the existence of black flies nationwide except around the capital and the coastal plain of Bonthe. In 1957, control efforts started with insecticides along the river breeding sites. In 1974, the Onchocerciasis Control Programme focusing on vector control was launched, extending to Sierra Leone in the 1980s. From 1995, the African Programme for Onchocerciasis Control commenced community-directed treatment with ivermectin (CDTI) in hyper- [microfilaridermia (mf) prevalence≥40%] and meso- (nodule prevalence 20%-39%) sites. To be effective programme, coverage among the eligible population over five years of age needed be at least 80%. In the mid-1990s, the rebel war stopped vector control and effective CDTI was not established until 2006. Baseline data using skin snips from 39 sentinel villages found the average OV mf prevalence was 53.1% (28.9 mf/snip). From October 2008, albendazole was added in phases to CDTI for all communities in all 14 districts for lymphatic filariasis (LF) elimination. In 2010, after five rounds of effective MDA, OV mf prevalence was 21.1% (8.29 mf/snip). Males had higher prevalence and density than females in both studies. Since then, OV control (acknowledging transmission continues) has transitioned to OV elimination assuming transmission can be interrupted once mf prevalence is less than 5%.

In 2017, a rapid assessment was integrated into LF-Transmission Assessment Survey and independently in 8 and 4 districts respectively, designed by the Survey Sample Builder. Children aged 5-9 years were randomly selected from 177 clusters and tested by OV-16 (Rapid Diagnostic Tests, SD BIOLINE) using finger-prick blood samples. Overall, 17 441 children participated and 347 (2.0%) were positive (M: 2.4% versus F: 1.6%) (P<0.001). All districts recorded low prevalence (range Bonthe: 0.9%-Kailahun: 3.0%) although Bombali had 5 of 12 hotspots (>10%). The LF- Transmission Assessment Survey sampling methodology does not take cognizance of black fly breeding sites, but did demonstrate ongoing transmission and another 4-5 rounds were advisable. A technical advisory committee has been established with representation from international experts to help plan for accreditation of onchocerciasis elimination around 2025.

Keywords: Onchocerciasis; Sierra Leone; Simulium damnosum; Microfilaridermia

Article history:

Received 26 August 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 An overview of chikungunya in Pakistan



Aisha Khan1, Sadia Selim Khan1, Muhammad Sohail Afzal2, Muhammad Asif Gondal1, Muhammad Isthiaq Ali3, Naveeda Riaz4, Majid Mehmood5, Haroon Ahmed1⊠

1Department of Biosciences, COMSATS University Islamabad (CUI), Islamabad, Pakistan

2Department of Life Sciences, University of Management & Technology (UMT), Lahore, Pakistan

3Department of Microbiology, Quaid-i-Azam University, Islamabad, Pakistan

4Department of Biotechnology, International Islamic University (IIUI), Islamabad, Pakistan

5Department of Zoology, University of the Poonch Rawalakot, AJK

⊠Corresponding author: Dr. Haroon Ahmed, Assistant Professor, Infectious Diseases Division,Department of Biosciences, COMSATS University Islamabad (CUI), Park Road, Chakh shahzad, Islamabad, Pakistan.

E-mail: haroonahmad12@yahoo.com; haroonahmed@comsats.edu.pk

Chikungunya is a viral disease which can infect human by bite of infected mosquitoes. This disease was initially reported in Tanzania. The symptoms of chikungunya are joint pain and fever. In Pakistan, cases were reported from different areas especially from Karachi. Many other cases of chikungunya were reported from Rawalpindi and Islamabad. Chikungunya cases were recorded from published data from Dec 2016 to September 2018 in different geographic regions of Pakistan. Chikungunya cases are being reported from many areas of Pakistan, but it is most common in Sindh. A total of 766 cases of chikungunya have been reported from all over Pakistan from Dec 2016 to September 2018. The prevalence of disease varied in different areas of Pakistan: FATA is 0%, Islamabad 4%, GB 0%, AJK 0%, Punjab 8%, Sindh 62%, KPK 22% and Baluchistan 4%. Mosquito breeding sites to human tenancy cause significant risk factor for dengue fever and chikungunya. There is no comprehensive treatment available of chikungunya in Pakistan. Generally speaking, antiviral drugs are administered during viral attack, but there is no antiviral drug for this dangerous disease. The WHO responds to chikungunya by providing guidance, technical support to many underdeveloped countries so they can effectively manage cases of chikungunya.

Keywords: Chikungunya; Incidence; WHO; Pakistan

Article history:

Received 10 September 2019

Revised 20 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 An overview of dengue fever in Pakistan



Aisha Khan1, Sadia Selim Khan1, Muhammad Asif Gondal1, Muhammad Sohail Afzal2, Muhammad Umer1, Muhammad Inam Afzal1, Muhammad Isthiaq Ali3, Majid Mehmood4, Haroon Ahmed1⊠

⊠Corresponding author: Dr. Haroon Ahmed, Assistant Professor, Infectious Diseases Division,Department of Biosciences, COMSATS University Islamabad (CUI), Park Road, Chakh shahzad, Islamabad, Pakistan.

E-mail: haroonahmad12@yahoo.com; haroonahmed@comsats.edu.pk

1Department of Biosciences, COMSATS University Islamabad (CUI), Islamabad, Pakistan

2Department of Life Sciences, University of Management & Technology (UMT), Lahore, Pakistan

3Department of Microbiology, Quaid-i-Azam University, Islamabad, Pakistan

4Department of Zoology, University of the Poonch Rawalakot, AJK

⊠Corresponding author: Dr. Haroon Ahmed, Assistant Professor, Infectious Diseases

Division,Department of Biosciences, COMSATS University Islamabad (CUI), Park Road,

Chakh shahzad, Islamabad, Pakistan.

E-mail: haroonahmad12@yahoo.com; haroonahmed@comsats.edu.pk

Dengue is a viral disease caused by bite of infected mosquitoes. The symptoms of this disease are headaches, pain behind eyes, nausea, flu, muscle pains, rash and swollen glands, etc. It can also develop into lethal disease known as severe dengue. The initial report of dengue fever in Pakistan was in 1994. It was seen sudden rise in cases of dengue fever. Dengue fever cases were recorded from published data from 2013 to August 2018 in different geographic regions of Pakistan. A total of 64 047 cases of dengue fever have been reported from all over Pakistan from period 2013 to August 2018. The prevalence of disease was varied in different areas of Pakistan: Fata is 1%, Islamabad 5%, AJK 0%, Punjab 19%, Sindh 22%, KPK 53% and Baluchistan 0%. Mosquito breeding sites and living in tropical areas or traveling within these areas may cause risk factor of this disease. The first dengue vaccine, Dengvaxia was recorded in several countries from 2015 to 2016. This was used for humans living in endemic areas. The WHO mentioned that countries should consider introducing of the dengue vaccine Dengvaxia only in areas where epidemiological data specify a high rate of disease.

Keywords: Dengue fever; Incidence; WHO; Pakistan

Article history:

Received 10 September 2019

Revised 20 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Long-effective and slowly released calcium cyanamide can inhibit mosquito growth and development: Implications for the prevention and control of dengue fever



Tian-le Tang1, Yi-ji Li1, Wang-yuan Wei2, Jia-gang Guo1⊠

⊠Corresponding author: Jia-gang Guo, Distinguished Professor, Tropical Medicine Research Institute of Hainan, Hainan Medical University, Haikou, Hainan, China.

E-mail: jiagangg@vip.163.com

1Hainan Medical University, Haikou, Hainan, China.

2The Institute of Parasitological Disease Prevention and Control of Hunan Province, Yueyang, Hunan, China

Dengue fever is one of the most serious mosquito-borne infectious diseases in the tropical world, causing substantial human morbidity and mortality, yet there is no specific treatment till now, and the main measures preventing dengue fever include applying pesticide to kill adult vector mosquitos and removing standing water to prevent the breeding of mosquitoes etc. Appropriate uses of such approaches can reduce dengue epidemic risk via reducing mosquito density. In fact, vector control using insecticides is the most common method to prevent and control mosquito-borne disease transmission. Nevertheless, recent research has shown that Aedes albopictus and Aedes aegypti, the two key vectors for dengue fever transmission, have rapidly developed high resistance to the most commonly used adult insecticide pyrethroid (deltamethrin and permethrin) and larvicide organophosphate (temephos). Therefore, it is of great significance to obtain a new drug with longer-term efficacy to prevent dengue virus transmission. Pesticides currently used act on adult stage of mosquitoes, typically applying to the environment where mosquito bites (on humans) occur, and very few target on egg or juvenile stages of the mosquitoes in the water environment. Calcium cyanamide (CaCN2), widely used as a fertilizer in many different settings, was recently shown to have potential to prevent tropical diseases by eliminating or inhibiting the growth and development of intermediate hosts. The main compounds of calcium cyanamide (CaCN2) are CaCN2 (50%- 60%) and calcium oxide (15%). The representative products of calcium cyanamide (CaCN2) are PERLKA® from Germany and ROEBON® from China. CaCN2 has recently been recognized to have the fungicidal effect on the pathogens of the soil borne diseases and addition of CaCN2 could benefit the thermophilic phase and the composting could quickly reach the sanitary standard and will contribute to solve the feces disposal problems. According to our team's long-term investigation and research, we found that ROEBON® (50% CaCN2) can be used to eliminate Oncomelania and effectively control the spread of schistosomiasis. In the latest research, we have confirmed that 10 mg/L CaCN2 (20 mg/L ROEBON®) can completely inhibit the emergence of mosquito larvae and the population growth from the perspective of inhibiting development and the efficacy can lasts for three months. Calcium cyanamide is a good candidate drug to control mosquito-borne infectious diseases transmission, especiallly for dengue fever prevention. In 2014, Scientific Committee on Health and Environmental Risks in European Commission promoted preliminary opinion on potential risks to human health and the environment from the utilization of calcium cyanamide as fertilizer, but convincing evidences for the causal connections between the exposure to CaCN2 and human health damages are missed. Given a 96-h acute zebrafish toxicity study, the No Observed Effect Concentration (NOEC) of calcium cyanamide to zebrafish was 100 mg/L CaCN2. In summary, most of the data are based on the evaluation of the effect of CaCN2 as a fertilizer, and the related research as a mosquito inhibitor is still very scarce. The research on toxicity of CaCN2 in mosquitoes needs to be further strengthened.

Keywords: Calcium cyanamide; Dengue fever; Mosquito

Article history:

Received 19 September 2019

Revised 23 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Updated status of cystic echinococcosis in humans and livestocks in Turkey



Sami Simsek1⊠, Harun Kaya Kesik2, Haroon Ahmed3

⊠Corresponding author: Sami Simsek, Department of Parasitology, Faculty of Veterinary, Firat University, Elazig, Turkey.

Tel.: +904242370000 (ext: 3967).

Fax: +904242388174

E-mail: ssimsek@firat.edu.tr

1Department of Parasitology, Faculty of Veterinary, Firat University, Elazig, Turkey

2Department of Parasitology, Faculty of Veterinary, Bingol University, Bingol, Turkey

3Department of Biosciences, COMSATS University Islamabad (CUI), Islamabad, Pakistan

Cystic echinococcosis has been frequently reported in different regions of Turkey. Based on the official announcements of the Ministry of Health, a total of 52 000 patients (3 257 patients per year) underwent surgery for cystic echinococcosis between 1990 and 2005 in the country. The annual incidence of cystic echinococcosis has been reported as 0.8-2.0/100 000, whereas the rate was supposed to be higher (6.4/100 000) in some regions. Although a total of 14 789 surgically confirmed cystic echinococcosis patients were reported between 2001 and 2005, the seroprevalence of human cystic echinococcosis has been estimated to be between 2.7%-14.6% in different parts of Turkey. A cross-sectional ultrasound-based study recorded abdominal cystic echinococcosis in 53 out of 8 618 healthy people screened in 2014-2015. There are many studies about molecular characterization of Echinococcus (E.) granulosus sensu lato in Turkey. It has been reported that E. granulosus sensu stricto (formerly G1-G3 genotypes) was the dominant species responsible for human cystic echinococcosis in Turkey. The G6 and G7 genotypes of E. canadensis also have been confirmed in human isolates. There have been many prevalence studies on cystic echinococcosis of farm animals in Turkey over the past years. Depending on the geographical area and specimens evaluated, the prevalence of cystic echinococcosis in livestock hosts ranged from 3.5% to 58.6%. However, the published studies showed that the prevalence of cystic echinococcosis could be ranged between 3.50%-70.91% in sheep, 1.60%-25.11% in goats, 3.00%-46.41% in cattle, and 10.24%-41.10% in buffaloes, respectively. A large part of genotyping studies at the molecular level, especially in livestock, have shown that E. granulosus s.s. (G1-G3 genotypes) is the most prevalent species responsible for cystic echinococcosis infection in sheep, cattle, goat, and camel. The G7 genotype of E. canadensis was found less frequently in sheep. Additionally, the G1 genotype of E. granulosus s.s and the G4 genotype of E. equinus have been confirmed in horses.

Keywords: Cystic echinococcosis; Human; Livestock; Turkey

Article history:

Received 13 September 2019

Revised 15 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Molecular identification of Strongyloides stercoralis infection in a severe pulmonary patient with pemphigus from Hainan Province of China



Li-hua Li1,2#, Jing-chen Xie3#, Jin-Long Chen1,2, Lang-yu Rao1,2, Li-xian Wu1,2, Hui Huang4, Gang Lu1,2,5⊠

#These authors contributed equally to this work.

⊠Corresponding authors: Gang Lu, Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, 571199, China.

E-mail: luganghn@163.com

Foundation project: This work was supported by the Innovation Research Team Project of Hainan Natural Science Foundation (2018CXTD340); the National Natural Science Foundation of China (No. 81672072 and No. 81760378); the Research and cultivation Fund Project of Hainan Medical College (HY2014-012).

1Departmentof Pathogenic Biology, Hainan Medical University, Haikou, Hainan, China

2Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, Hainan 571199, China

3Department of Respiratory Medicine of Hainan West Central Hospital, Danzhou City of Hainan Province 571700, China

4Laboratory Department of Haikou People's Hospital, Haikou, Hainan, China

5Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China

Objective: To describe a case of Strongyloides (S.) stercoralis infestation with severe lung involvement in a patient who had pemphigus and treated with prednisone and to analyze the molecular characteristics of the S. stercoralis isolate.

Methods: A 54-year-old Chinese man living in rural area of the Hainan Province, China. He had a 1-year history of pemphigus and took prednisone for more than six month. He was admitted to the hospital with a 1-week history of abdominal pain, nausea and vomiting. Two days before admission, he presented with cough, expectoration, and shortness of breath after activity accompanied by fever. Computed tomography (CT) of the chest was performed at different stage of admission. Gastric juice, sputum and stool sample smears were examined under light microscope. The larvae of Strongyloides were found from gastric juice, sputum and stool samples of the patient and the genomic DNA of the larvaes was extracted. PCR amplification of the cytochrome c oxidase subunit 1 (cox1) gene was carried out for the DNA sample and the products were sequenced.

Results: CT of the chest on day 1 of admission showed inflammation in the tongue segment of the upper lobe of the left lung and the lower lobe of both lungs and the subsequent CT of the chest revealed extensive bilateral pulmonary infiltrates. PCR amplification and sequencing analysis of the Strongyloides larvae directly colleted from the gastric juice, sputum and stool samples on day 10 of admission showed that 100% homology with the sequence of S. stercoralis isolated in a human from Iran (GenBank No. MG995852). Clinical and imageologicalimprovement was achieved after treatement of albendazole (600 mg/day).

Conclusions: This is the first case report of pulmonary S. stercoralis infection in a patient with pemphigus treated with prednisone in Hainan Province of China. We suggested that strongyloidiasis should be routinely investigated in patients presenting with nausea, vomiting, diarrhea, cough or expectoration which were accompanied by fever, especially for patients with low immune function or immune deficiency.

Keywords: Pulmonary; Strongyloides stercoralis; Pemphigus; Hainan

Article history:

Received 5 September 2019

Revised 25 September 2019

Accepted 26 September 2019

Available online 7 October 2019

 Melioidosis: A hidden tropical disease in Sri Lanka



Enoka Marie Corea

⊠Corresponding author: Dr. Enoka Marie Corea, Faculty of Medicine, University of Colombo, Sri Lanka.

E-mail: enokacorea@hotmail.com

Faculty of Medicine, University of Colombo, Sri Lanka

Melioidosis, caused by the soil saprophyte Burkholderia pseudomallei, is found in tropical countries. Infection is acquired from soil and water by inoculation through the skin and mucous membranes, inhalation into the respiratory tract and ingestion of untreated water. Clinical manifestations range from acute fulminant septicaemia to chronic localized infection characterised by abscess formation. Isolation of Burkholderia pseudomallei from patient specimens confirms the diagnosis. Although exposure is widespread in rural, agricultural communities, clinical disease is less common as it usually requires compromised immunity, commonly diabetes. Case fatality is high in the absence of effective antibiotic treatment which comprises an acute phase of intravenous antibiotics and a long eradication phase of oral antibiotics. Prevention and control is difficult as the bacterium is widespread in the soil and water of endemic countries and exposure is common in rural, agricultural populations. Sri Lanka was not considered a country with endemic melioidosis in the past. National surveillance was instituted in 2006. Around 450 culture positive cases and more than 1 000 antibody positive cases were recorded to date. Males predominated. The age range was wide. All provinces were affected with the majority living in rural areas. Case load increased during the two monsoons. There was representation of all population groups including farmers, drivers, housewives, school children, professionals, businesspersons, white collar and blue collar workers. Diabetes was the predominant risk. Clinical presentations included community acquired sepsis and pneumonia, superficial and deep abscesses, septic arthritis, neurological melioidosis and endocarditis. Mortality was more than 20%. Melioidosis is endemic in Sri Lanka with a wide geographic and demographic distribution. There is a need to extend surveillance to under-resourced parts of the country.

Keywords: Melioidosis; Sri Lanka; Epidemiology

Article history:

Received 2 September 2019

Revised 24 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Evaluation of the human-baited double net trap and BG trap as an alternative to human landing catches for collecting outdoor Aedes albopictus in China



Yu-yan Wu, Qin-mei Liu, Jin-na Wang, Tian-qi Li, Zhen-yu Gong, Juan Hou⊠

⊠Corresponding author: Hou Juan, Zhejiang Provincial Center for Disease Control and Prevention, China.

Tel.: 13958113504

E-mail: jhou@cdc.zj.cn

Zhejiang Provincial Center for Disease Control and Prevention, China

Objective: Monitoring the density of Aedes(Ae.) albopictus was very important in evaluating the efficiency of vector management, especially when Aedes-borne diseases break, like Dengue fever, Chikungunya fever and Zika. Human landing catch (HLC) is regarded as the “gold standard” for Aedes monitoring, but it is unsafe and unethical since it potentially expose field professionals to a series of pathogens. In this study, we compared HLC with human-baited double net (HDN) and BG trap for field Aedes albopictus monitoring.The study aims to find an effective and safe alternative method to HLC in monitoring Ae. albopictus, especially in emergency monitoring.

Methods: Latin square design was used, and three sites in Hangzhou city, Zhejiang province, China, were chosen to conduct outdoor HLC, HDN and BG trap catches in June. The tests were performed at three periods: a morning period (8:30-10:00), an afternoon period (15:00-16:30), and an evening period (16:30-18:00). Then a table with 81 elements was made to compare the efficiency of these three methods.

Results: A total of 80, 138 and 78 adult Ae. albopitus was captured by HDN, BG trap and HLC, respectively. Among these three catches, BG trap showed the best efficacy in captured Ae. albopitus adults. The mosquitoes caught by HLC and BG trap were nearly the same at all three time series, but the density of mosquitoes caught by HDN at 16:30-18:00 was 2 times the density at 8:30-10:00 (4.44 vs 2.22 per hour). In this study, significant positive correlation between HLC and BG trap of Aedes albopitus female density was found (r=0.921, P<0.001). However, no statistical significant correlation between HLC and HDN, BG trap and HDN of Ae. albopitus female density was found (r=-0.46, P= 0.820; r=0.019, P=0.923, respectively). Besides, the human-bait attraction bias of HLC catches might be more apparent than BG trap and HDN catches.

Conclusions: According to our study, with high efficiency, less human-bait attraction bias, and significantly positive correlation with HLC in catching Ae. albopitus females, BG trap could be a sensitive and safer alternative to HLC for outdoor Ae. albopitus monitoring. It also showed strong potential in emergency monitoring when Aedesborne diseases break.

Keywords: Aedes albopictus; Human landing catch; Human-baited double net trap; BG trap; Monitoring

Article history:

Received 17 September 2019

Received in revised form 26 September 2019

Accepted 30 September 2019

Available online 7 October 2019

 An overview of Crimean-Congo hemorrhagic fever in Pakistan



Sadia Selim Khan1, Aisha Khan1, Muhammad Sohail Afzal2, Shahzad Ali3, Haroon Ahmed1⊠

⊠Corresponding author: Dr. Haroon Ahmed, Assistant Professor, Infectious Diseases Division, Department of Biosciences, COMSATS University Islamabad (CUI), Park Road, Chakh shahzad, Islamabad, Pakistan.

E-mail: haroonahmad12@yahoo.com; haroonahmed@comsats.edu.pk

1Department of Biosciences, COMSATS University Islamabad (CUI), Islamabad, Pakistan

2Department of Life Sciences, University of Management & Technology (UMT), Lahore, Pakistan

3Department of Fisheries & Wildlife, University of Veterinary and Animal Sciences(UVAS), Lahore Pakistan

Crimean-Congo Hemorrhagic Fever (CCHF) is a disease affecting domestic livestock and wild animals which can spread to humans. It is caused by infection with a tick- borne virus (Nairovirus) in the family Bunyaviridae or by contacting with infected tissues or from animal blood. CCHF cases were recorded from published data from 2013 to 2018 in different geographic regions of Pakistan. The intensity and risk factors were also determined from all four provinces of Pakistan. A total of 391 cases of CCHF have been reported from all over Pakistan during period of 2013-2018. Majority of them were recorded at the time of Eid-ul-Adha. CCHF cases were identified predominantly in Baluchistan (n=12), Karachi (n=5), Bahawalpur (n=2), and Khyber Pakhtunkhwa (n=1). The prevalence of disease were different in different areas of Pakistan (Fata 1%, Islamabad 5%, Punjab 21%, Sindh 8%, KPK 14% and Baluchistan 39%). The political disturbances faced by the Pakistan have increased Pakistan's susceptibility because large number of refugees have migrated to Pakistan from Afghanistan which is an endemic country. Most of the immigrants and their cattles from Afghanistan settled in Khyber, Pakhtunkhwa and Baluchistan provinces which ultimately cause higher prevalence of CCHF in these arears. Currently there is no complete cure or commercially available vaccine of CCHF available in Pakistan. Mostly Ribavirin antiviral drug is used to treat CCHF. The disease can be controlled by implementing preventive measures like avoiding contact with blood of the suspected animal and tick bites.

Keywords: Crimean Congo haemorrhagic fever; Eid-ul-Azha; CCHF; Pakistan

Article history:

Received 19 September 2019

Revised 22 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Epidemiology and clinical features of scrub typhus in Hainan, China



Gao-yu Wang1,2#, Li-yuan Zhang3#, Shu Wen4#, Wen-qi Wang1,2, Xiao-yuan Hu1,2, Ruo-yan

Peng1,2, Yi Huang1,2, Zhang-li Lin4, Kun-liang Wu3, Gang Lu1,2,5, Jasper Fuk-woo Chan1,6,7,8, Long Sun9, Kwok-yung Yuen1,6,7,8, Yong-guo Du3⊠, Fei-fei Yin1,2⊠

#These authors have contributed equally to this work

*Corresponding author: Fei-fei Yin, Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan Province, China.

E-mail: yinfeifeiff@163.com

Yong-guo Du, Department of Infectious Diseases, the Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China.

E-mail: duyongguo@163.com

Foundation project: This study was partly supported by the National Natural Science Foundation of China (Grant No. 81860367), and State Key Laboratory of Virology 2018 Open Fund Project (2018IOV002).

1Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan Province, China

2Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, Hainan Province, China

3Department of Infectious Diseases, the Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China

4Clinical laboratory, People's Hospital of Qiongzhong Li Miao Autonomous County, Haikou, Hainan Province, China

5Department of Pathogen Biology, Hainan Medical University, Haikou, Hainan Province, China

6State Key Laboratory of Emerging Infectious Diseases, the University of Hong Kong, Pokfulam, Hong Kong

7Department of Microbiology, the University of Hong Kong, Pokfulam, Hong Kong

8Carol Yu Centre for Infection, the University of Hong Kong, Pokfulam, Hong Kong

9Department of Infectious Diseases, the first Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China

Objective: To identify the epidemiology and clinical features of scrub typhus in Hainan Province, China.

Methods: A total of 258 patients with acute febrile illness were recruited from four hospitals in Hainan between 1st June 2018 and 31th August 2019. Orientia tsutsugamushi-specific IgM/IgG and DNA were detected by goldimmuno- chromatographic assay and PCR on the patients' serum samples. The clinical and epidemiologicalcharacteristics of the laboratory-confirmed cases were recorded using a predesigned database.

Results: The positive rates of IgM and IgG were 22.5% (58/258) and 39.9% (103/258), respectively, and the positive rate of PCR was 19.37% (50/258). There were 55 males and 48 females among the 103 IgG-positive patients. Patients of all age groups were affected (10 to 86 years); 65.0% of the adults aged 41 to 70 years (67/103); 64.1% of the patients were farmers (66/103). The patients resided in 12 counties and cities of Hainan, and the highest number of cases being reported among residents in Qiongzhong County and Haikou City (54/103, 52.4%). Seasonal variations in the incidence rate of scrub typhus was observed, with the peak incidence (53/103, 51.5%) occurring in the summer (June to August). The common presenting symptoms were fever (103/103, 100.0%), eschar or skin ulcer (38/103, 36.9%), and chills and rigors (30/103, 29.1%). The major complications were liver dysfunction (9/103, 8.7%). Four genotypes of Orientia tsutsugamushi were identified among these patients, including Karp, Kato, Gilliam and Ta763. The predominant genotype was Karp (33/51, 64.7%).

Conclusions: Scrub typhus is endemic in Hainan, which predominantly affects residents with prolonged outdoor activities such as farming.

Keywords: Scrub typhus; Orientia tsutsugamushi; Hainan; Epidemiology

Article history:

Received 19 September 2019

Recvised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Prevalence and determinants of childhood obesity in Haikou, China: A cross-sectional study



Fan-xian Ou-yang1⊠, Liang-liang Yang2, Hui-lan Xu3, Ren-he Yu4, Ping-feng Bu5

⊠Corresponding author: Fan-xian Ou-yang, Department of Preventive Medicine, the Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China.

Tel.: 086-0898-66809196

E-mail: hnnkoyfx@aliyun.com

Foundation project: It was supported by Hainan Applied Technology Research and Development Special Project (No. ZDXM2014063) and Hainan Social Development Science and Technology Project (No. SF201313).

1Department of Preventive Medicine, the Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China

2XiangYa School of Public Health, Central South University, Changsha, Hunan Province, China

3Department of Social Medicine and Health Service Management, XiangYa School of Public Health, Central South University, Changsha, Hunan Province, China

4Epidemiology and Health Statistics, XiangYa School of Public Health, Central South University, Changsha, Hunan Province, China

5Department of Pathology, the Second Affiliated Hospital of Hainan Medical College, Haikou, Hainan Province, China

Objective: To identify the prevalence and factors of rapidly increasing childhood obesity in Haikou, China.

Methods: A total of 11 497 students from 227 classes in primary and secondary schools of Haikou were surveyed by questionnaires from October 2013 to December 2014 using the Stratified Random Cluster Sampling method. Height and weight of the students were measured. A total of 8 458 children aged 7-15 years old were selected for the prevalence analyzing of overweight/obesity and 7 836 for logistic stepwise regression analysis.

Results: The rate of overweight and obesity were 10.5% and 7.4%, respectively. Out of the 33 investigated factors, 17 were associated with childhood obesity (P<0.05), including 9 risk factors (OR>1) and 8 beneficial factors (OR<1). Birth weight (macrosomia), parent's obesity are the top three risk factors and children's breakfast hobby (soybean milk) and food taste (sweet), favorite diet style of the family (stewing) are beneficial factors.

Conclusions: The overweight/obesity rate is already at a high level and early interventions are needed. The necessary tasks are to strengthen education on parents and children with prevention knowledge, appropriately reduce the learning burden, and control the average time of watching TV to less than 3 h a day.

Keywords: Childhood obesity; Risk factor; Tropical region; Haikou city

Article history:

Received 19 August 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Analysis of determinants of suicide ideation among Hainan medical students based on structural equation model



Xi-wen Liu, Ling Wu⊠, Xiao-dan Wang⊠, Wei-ling Xu

⊠Corresponding author: Ling Wu, School of Public Health, Hainan Medical University, Haikou Hainan 571199, China.

E-mail: 507816301@qq.com

Xiao-dan Wang, School of Public Health, Hainan Medical University, Haikou Hainan 571199, China.

E-mail: xiaodanwangkk@126.com

School of Public Health, Hainan Medical University, Haikou Hainan 571199, China

Objective: To investigate suicide status and influence of medical students in Hainan, and analyze the direct and indirect effects of influencing factors on suicide ideation among college students based on the structural equation mode1.

Methods: By using multi-stage sampling method, we selected 696 students of 11 majors of Hainan Medical College as the research subjects, by questionnaire, simplified coping style questionnaire, Adolescent Self-Rating Life Events Scale (ASLEC), self-rating depression scale (SDS), social support rating scale, the self-esteem inventory and the Self-rating Idea of Suicide Scale(SIOSS) was conducted.

Results: The idea of suicide in Hainan Medical College students occurred in 15.4%, and suicide act ideation rate was higher in male than in female (χ2=7.161, P = 0.006), non-medical higher medical (χ2=6.004, P =0.014). The goodness of fit for the structural equation model was satisfactory and major indices had met corresponding requirements. The depression was a direct influencing factor on suicide ideation, while three factors including social support, positive coping style and passive coping style had indirect impacts.Negative life event and selfesteem not only directly affected suicide ideation, but also had indirect effects.According to the percentages of their contribution, the risk factors were ranked as follows: depression (45.6%), negative life event (33.6%) and passive coping style (18.5%). Similarly, the top protective factors were self-esteem (37.0%), followed by social support (28.4%) and positive coping style (14.3%).

Conclusion: The higher incidence of suicide idea of students at the Hainan Medical College, depression and selfesteem were the main factors of suicide ideation. So, it is of great importance to not only strengthen the mental health of college students, but also to train students to face up the difficulties with a positive style, and provide the college students with sufficient social supports.

Keywords: Structural equation model; Suicide ideation; Factor analysis; Medical students; Hainan

Article history:

Received 17 September 2019

Received in revised form 26 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Survey of vitamin A nutrition status of primary and middle school students in poverty-stricken areas in tropical region from 2014 to 2017



Li Cao, Bing Zhang , Xiao-dan Wang⊠

⊠Corresponding author: Wang Xiao-dan, School of Public Health, Hainan Medical University, Haikou, Hainan, 571199, China.

E-mail: 1037822186@qq.com; 794804246@qq.com

Foundation project: This study is supported by National Natural Science Foundation of China (No. 81860577).

School of Public Health, Hainan Medical University, Haikou, Hainan, 571199, China

Objective: To understand the levels and intake of dietary vitamin A by the primary and secondary school students in poverty-stricken areas in tropical region, and provide suggestions for the follow-up implementation of students' nutrition improvement.

Methods: A multi-stage cluster sampling method was adopted to extract 3 junior high schools and 3 primary schools in Qiongzhong county of Hainan province, and students of 1 or 2 classes were selected as subjects for investigation. Fasting venous blood was collected. The level of serum vitamin A (retinol) was determined by high performance liquid chromatography. Excel 2007 and SPSS 20.0 software were used for the one-way ANOVA and χ2 tests.

Results: The vitamin A deficiency rates from 2014 to 2017 were 1.6%, 1.5%, 2.0%, and 0.8% respectively, and the difference was not statistically significant (χ2=3.544, P =0.31 > 0.05). The subclinical deficiency rates of vitamin A were 22.3%, 31.6%, 27.9%, 18.0% respectively, and the difference was statistically significant (χ2 = 40.240, P =0.00 < 0.05). There was no statistically significant difference between the genders in vitamin A deficiency and vitamin A subclinical deficiency from 2014 to 2017. The subclinical deficiencies of primary vitamin A in primary school students were higher than those in middle school students (P<0.05).

Conclusion: The nutritional status of vitamin A in primary and secondary school students of the “Student Nutrition Improvement Program” in Hainan Province has been significantly improved, and this program can be recommended in the whole province.

Keywords: Poverty area; Student; Vitamin A; Nutritional status

Article history:

Received 17 September 2019

Rvised 25 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Analysis of 8 970 pregnant women with infectious diseases in Haikou, China



Jie-ying Wang, Zi-hua Tong, Chun-chun Yu, Yu-cheng Li, Yan-qun Xie, Hua Pei⊠

⊠Corresponding author: Hua Pei, the Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China.

E-mail: phzmh61@aliyun.com

The Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China

Objective: To preliminarily investigate the infection status of the human immunodeficiency virus (HIV), hepatitis B virus (HBV), treponema pallidum and hepatitis C virus (HCV) in pregnant women in Hainan Province, so as to provide evidence for effective prevention and treatment of mother-to-child transmission.

Methods: From October 2017 to July 2019, 8 970 pregnant women who came to our hospital for outpatient examination during pregnancy were tested for HIV, treponema pallidum and HCV antibody by ELISA and tested for rapid detection of HBV surface antigen in serum by gold immune-chromatographic assay.

Results: The positive rate of HIV, treponema pallidum and HCV antibody was 0.01% (1/8 970), 0.65% (58/8 970) and 0.11% (10/8 970), respectively, and the positive rate of HBV surface antigen was 11.13% (998/8 970).

Conclusions: Among the 8 970 pregnant women, the HBV infection rate was the highest (11.13%), significantly higher than that of the other three infectious diseases, followed by the treponema pallidum and HCV positive rate, and the HIV positive rate was the lowest. Therefore, it is suggested that pregnant women should strengthen prenatal examinations to improve the awareness of prevention of infectious diseases, which is conducive to prepotency. Meanwhile, it is urgent to improve the screening and treatment for epidemic infectious disease in Haikou.

Keywords: Pregnant women; HIV; HBV; Treponema pallidum; HCV

Article history:

Received 4 September 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Clinical effectiveness of live preparation of lactobacillus in treatment of bacterial vaginosis in pregnancy: A meta-analysis



Xiao-zhen Li, Mei-jiao Li, Ye-feng Wang, Hai-rong Huang, Wen-ting Cao⊠

⊠Corresponding author: Wen-ting Cao, School of Public health, Hainan Medical University, Haikou, Hainan Province, China.

E-mail: 1205969243@qq.com

Foundation project: It was supported by Research Foundation of Hainan Medical University (No. HY2018-14).

School of Public health, Hainan Medical University, Haikou, Hainan Province, China

Objective: To evaluate the clinical effectiveness of live preparation of lactobacillus in treatment of bacterial vaginosis in pregnancy.

Methods: Randomized controlled trials of live preparation of lactobacillus in the treatment of bacterial vaginosis in pregnancy were collect by searching PubMed, Web of Science, OVID, CNKI, WanFang, VIP, CBM and Elsevier databases. Quality of the included trials were evaluated by two researchers independently, and data were extracted according to Cochrane systematic evaluation. RevMan 5.3 software was used for meta-analysis.

Results: Twenty-one randomized controlled trials involving 2 930 patients were included, which showed that there was significant difference in the clinical effectiveness between vaginal medication of live preparation of lactobacillus and vaginal medication of metronidazole [total effective rate (RR=1.05, 95% CI: 1.02-1.07, P=0.000 4]; significant differences were found in premature delivery rate (RR=0.49, 95% CI: 0.32-0.73, P=0.000 4), premature rupture of membrane rate (RR=0.54, 95% CI: 0.38-0.77, P=0.000 7), infant of low-birth weight rate (RR=0.45, 95%CI: 0.22-0.94, P=0.03), puerperal infection rate (RR=0.60, 95% CI: 0.39-0.94, P= 0.03) between the two groups.

Conclusions: Vaginal medication of live preparation of lactobacillus was more clinically effective than vaginal medication of metronidazole for bacterial vaginosis in pregnancy. Live preparation of lactobacillus is associated with a lower premature delivery rate, a lower premature rupture of membrane rate, a lower low-birth weight rate and a lower puerperal infection rate.

Keywords: Enterocytozoon bieneusi; Genotype; ITS region; Macaca; Zoonotic

Article history:

Received 6 September 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Molecular identification of the first case of aural myiasis caused by Sarcophaga peregrina in an infant from Hainan, China



Gang Lu1,2,3#, Wei Zhao1,2,3#, Huan-huan Zhou1,2,3, Mei-fang Xiao4, Yong-juan Yu5, Hai-fen Nong4, Wei Xiang4⊠

#These authors contributed equally to this work.

⊠Corresponding author: Wei Xiang, Hainan Women and Children's Medical Center, Haikou, Hainan Province, China.

E-mail: xiangwei8@163.com

Foundation project: This work was supported by the Innovation Research Team Project of Hainan Natural Science Foundation (No. 2018CXTD340) and the National Natural Science Foundation of China (No. 81672072 and No.81760378).

1Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan Province, China

2Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan Province, China

3Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, Hainan Province, China

4Hainan Women and Children's Medical Center, Haikou, Hainan Province, China

5Laboratory Department of Baisha People's Hospital, Yacha, Hainan Province, China

Objective: To report a case of aural myiasis in an infant and molecular identification of larva species.

Methods: A larva was extricated from left external auditory canal of an infant in the Neonatal Department of the People's Hospital of Baisha County, Hainan Province on 25 June 2019. The larva was placed in 70% alcohol and sent to the Department of Pathogenic Biology, Hainan Medical University. The morphological characters of larva was observed under stereoscopic microscope. The genomic DNA of the larva was extracted and amplified by PCR targeting mtDNA cytochrome c oxidase subunit 1 gene of Diptera, then the PCR product was sequenced and analyzed.

Results: A 5-day-old male infant with neonatal hyperbilirubinemia became suddenly irritable. Physical examination found bleeding of the left external auditory canal. A larva about 1.0 cm×0.2 cm was found in the left external auditory canal when the left ear was cleaned with saline. The infant was transferred to Hainan Women and Children's Medical Center for further examination. Otoscopic examination found that the right external auditory canal was normal, with a small amount of yellow secretion and intact tympanic membrane. However, the left ear canal was slightly swollen, with a small amount of yellow secretion and no obvious perforation of the tympanic membrane. The child was discharged after four days of treatment with appropriate medication. Morphological characteristics observed by stereoscopic microscope implied that larva was the maggot of Sarcophaga sp. PCR amplification and sequencing analyses confirmed that the larva was Sarcophaga (S.) peregrina. The sequence exhibited 100% homology with the S. peregrina c oxidase subunit 1 gene (GenBank No. AF259509.1).

Conclusions: This is the first report of an infant with aural myiasis in Hainan Province. The molecular characteristics suggest that the aural myiasis was caused by S. peregrina. One of the factors causing infant arual myasis might be residual amniotic fluid in the external auditory canal of the newborn.

Keywords: Myiasis; Sarcophaga peregrina; Infant; Molecular; Hainan

Article history:

Received 5 September 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Molecular identification and subtype analysis of Blastocystis in pigs, cattle and goats in Hainan, China



Wei Zhao1,2,3, Tian-ming Ma1,2,3, Huan-huan Zhou1,2,3, Meng Qi4, Zong-xi Cao5, Zhe Chao5, Li-min Wei5, Quan-wei Liu5, Rui-ping Sun5, Feng Wang5, Yan Zhang5, Xin-li Zheng5⊠, Gang Lu1,2,3⊠

⊠Corresponding author: Gang Lu, Department of Pathogenic Biology, Hainan Medical University, No. 3 Xueyuan Road, Haikou, Hainan Province, China.

E-mail: luganghn@163.com

Xin-li Zheng, Institute of Animal Science and Veterinary Medicine, Hainan Academy of Agricultural Sciences, Haikou, Hainan Province, China.

Foundation project: This work was supported by the Innovation Research Team Project of Hainan Natural Science Foundation (No. 2018CXTD340); the National Natural Science Foundation of China (No. 81672072 and No.81760378); the Young Talents Science and Technology Innovation Project of Hainan Association for Science and Technology (No. QCXM201802); the Hainan major science and technology project (No. ZDKJ2016017-01).

1Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, China

2Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China

3Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, Hainan Province, China

4College of Animal Sciences, Tarim University, Alar, Xinjiang Province, China

5Institute of Animal Science and Veterinary Medicine, Hainan Academy of Agricultural Sciences, Haikou, Hainan Province, China

Objective: To determine the infection rates and subtype distributions of Blastocystis in pigs, cattle, and goats in Hainan Province, China.

Methods: From September 2018 to May 2019, a total of 659 fresh fecal samples were collected from 188 pigs, 341 goats, and 130 cattles farmed in cities of Hainan Province. All the samples were analyzed for the presence of Blastocystis by amplifying the SSU rRNA gene. Phylogenetic analysis was performed by constructing a neighborjoining tree using the program Mega X.

Results: Thirty-one percent (204/659) of the animals were confirmed to be infected with Blastocystis, with 42.2% (144/341) of the goats, 30.8% (40/130) of the cattle, and 10.6% (20/188) of the pigs. Twelve subtypes of Blastocystis were identified, including ST10 (n=114), ST5 (n=37), ST26 (n=16), ST24 (n=8), ST14 (n=8), ST21 (n=8), ST23 (n=4), ST25 (n=4), ST1 (n=2), ST2 (n=1), ST4 (n=1), and ST12 (n=1). There were 11 (ST1, ST2, ST4, ST5, ST10, ST12, ST14, ST21, ST23, ST24, and ST26) and six subtypes (ST10, ST21, and STs23- ST26) in goats and cattle, respectively. All the pig-derived Blastocystis isolates belonged to ST5. A total of 37 representative sequences were obtained from the 12 subtypes of Blastocystis, including 14 sequences that have been described previously and 23 novel sequences were identified which were composed of ST10 (n=9), ST5 (n=8), ST21 (n=3), ST26 (n=2), and ST14 (n=1).

Conclusions: This is the first study to identify the presence of Blastocystis in pigs, cattle, and goats from Hainan Province, China, and provides baseline data useful for controlling and preventing Blastocystis infection in farm communities. Pigs and goats appear to be the major reservoirs of potential zoonotic infections with ST1, ST2, ST4, ST5, and ST12 and zoonotic infections in pigs and goats with those subtypes should be considered potential public health threats. The new ST sequences of Blastocystis identified here provide novel insights into the genotypic variation in Blastocystis.

Keywords: Blastocystis; Pig; Goat; Cattle, Genotypic variation

Article history:

Received 5 September 2019

Revised 13 September 2019

Accepted 25 September 2019

Available online 7 October 2019

 Emerging trends of theileriosis in Pakistan



Sadia Selim Khan1, Aisha Khan1, Nazeer Hussain1, Muhammad Sohail Afzal2, Muhammad Asif Gondal1, Haroon Ahmed1⊠

⊠Corresponding author: Dr. Haroon Ahmed, Assistant Professor, Infectious Diseases Division, Department of Biosciences, COMSATS University Islamabad (CUI), Park Road, Chakh shahzad, Islamabad, Pakistan.

E-mail: haroonahmad12@yahoo.com; haroonahmed@comsats.edu.pk

1Department of Biosciences, COMSATS University Islamabad (CUI), Islamabad, Pakistan

2Department of Life Sciences, University of Management & Technology (UMT), Lahore, Pakistan

Theileriosis is a disease caused by protozoan parasites which belong to genus Theileria. It can be transmitted through ixodid ticks. Mostly the distribution of Theileria species are restricted to subtropical and tropical regions. Theileria species cause economically significant diseases in cattle, sheep, and goats in Pakistan by infecting domestic and wild ruminants. This is a neglected disease in Pakistan because limited research work has been done yet on it. Ovine theileriosis cases were recorded from published data from 2013-2016 in different geographic regions of Pakistan. The intensity and risk factors were also determined in KPK Province of Pakistan. Theleriosis have been reported from all over Pakistan during period 2013-2016. The prevalence of Theleria annulata in buffaloes ticks was 72.91% (875) in urban regions of Hyderabad and it was 80.8% (970) in periurban regions during period 2013-2014. In 2016, prevalence of ovine theileriosis in KPK districts was as 27.5% in Dera Ismail Khan, 13% in Tank and 19.5% in Bannu. It is concluded that paying close attention to animal keeping, feeding and housing may diminish the occurrence of theileriosis in sheep. The higher prevalence might be due to lack of awareness by the community, farming system of the rural livestock of, absence of diagnostic facilities, expensive acaricidal drugs and poor economic condition of farmers.

Keywords: Theileriosis; Status; Outbreak; Pakistan

Article history:

Received 20 September 2019

Revised 22 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Distribution and molecular characterization of Cryptosporidium species or genotypes in four species of farmed animals from Hainan Island of China



Tian-ming Ma1,2,3#, Huan-huan Zhou1,2,3#, Hai-ju Liu1,2,3, Jing-guo Zhou1,2,3, Xin-li Zheng4, Meng Qi5, Gang Lu1,2,3⊠, Wei Zhao1,2,3,6⊠

#These authors contributed equally to this work.

⊠Corresponding author: Wei Zhao, Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, 571199, China.

E-mail: hayidazhaowei@163.com

Gang Lu, Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, 571199, China.

E-mail: luganghn@aliyun.com; ganglu2018@163.com

Foundation project: This work was supported by the Innovation Research Team Project of Hainan Natural Science Foundation (2018CXTD340); the National Natural Science Foundation of China (No. 81672072 and No.81760378); the Open Subject of Key Laboratory of Parasite and Vector Biology, MOPH (WSBKFKT-201802).

1Department of Pathogenic Biology, Hainan Medical University, Haikou, Hainan, China

2Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China

3Key Laboratory of Translation Medicine Tropical Diseases, Hainan Medical University, Haikou, Hainan, 571199, China

4Institute of Animal Science and Veterinary Medicine, Hainan Academy of Agricultural Sciences, Haikou 571100, China

5College of Animal Science, Tarim University, Alar, Xinjiang, 843300, China

6Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China

Objective: To assess the infection status of Cryptosporidium spp. in the four common species of farmed animal (pigs, cattle, goats and geese) in the Hainan Island of China, followed by molecular characterization of the parasite.

Methods: During March to July 2019, 864 DNA were extracted from the fecal samples of 188 pigs, 341 goats, 130 cattle, and 169 geese which were collectd from four, five, six, and three cities of the Hainan Province, China, respectively. The species/genotypes and subtypes of Cryptosporidium were identified by employing nested-PCR amplification of the SSU rRNA and the 60 kDa glycoprotein (gp60) genes of the parasite, respectively.

Results: Cryptosporidium was identified in 68 (7.9%) out of 864 animals, with 45 (13.2%) goats, 13 (6.9%) pigs, 4 (3.1%) cattle, and 6 (3.6%) geese. Cryptosporidium (C.) xiaoi (n=24) and C. ubiqutium (n=21) were identified in goats; C. suis (n=7) and C. scrofarum (n=6) in pigs; C. parvum (n=3) and C. occultus (n=1) in cattle; and C. baileyi (n=1) and Cryptosporidium goose genotype I(n=3) in geese. The 21 C. ubiqutium isolates shared a same subtype XIIa, and the subtype of three C. parvum was II dA15G1.

Conclusions: We present for the prevalence of the Cryptosporidium in cattle, goats, pigs and geese from the Hainan Island of China for the first time. The molecular characterization of the Cryptosporidium isolates obtained here propose that the goats, cattle and pigs are potential sources of C. ubiqutium, C. parvum, and C. suis infections, in humans.

Keywords: Cryptosporidium; Farmed animals; Zoonotic; Hainan Island

Article history:

Received 20 September 2019

Revised 22 September 2019

Accepted 24 September 2019

Available online 7 October 2019

 Sub-lethal dose of Bacillus sphaericus suppresses vector competence of Anopheles dirus for Plasmodium yoelii by up-regulating the Imd signaling pathway



Sha-sha Yu#, Jie Qin#, Qiu-xia Li, Pan Wang, Jing Wang, Ting-ting Liu, Ying Wang⊠

#These authors contributed equally to this work.

⊠Corresponding author: Ying Wang, Ph.D., Department of Tropical Medicine, College of Military Preventive Medicine, Army Medical University, Chongqing, 400038, China.

Tel.: +086 02368771576

E-mail: wangyingtmmu2016@126.com

Department of Tropical Medicine, College of Military Preventive Medicine, Army Medical University, Chongqing, 400038, China

Objective: To investigate the effect of sublethal dose of Bacillus sphaericus (Bs) 2 362 strain on the malaria transmission ability of Anopheles (An.) dirus (Hainan strain) and its molecular mechanism.

Methods: Firstly, the fourth instar larvae of An. dirus was treated with sublethal dose of Bacillus sphaericus. The surviving larvae were then collected and placed into mosquito cages, where they were hatched into adult mosquitoes. These mosquitoes were recorded as Bs group. Meanwhile, the control group was set without any treatment. Then, for infection with Plasmodium yoelii BY265 RFP, 3- to 5-day old female adults were kept at 24 °C and fed on Plasmodium yoelii BY265 RFP-infected Kunming mice with a gametocytemia above 0.5%. On day 9-11 post infection, mosquitoes were dissected, and the oocysts on the midguts were examined under a fluorescence microscope. Thirdly, total RNA was extracted from mosquitoes of Bs group and the control group at different time-points respectively, and the cDNA were synthesized later. Finally, SYBR quantitative PCR was conducted to investigate the expression of Imd pathway anti-malaria molecules at different time-points, including TEP1 and Rel2, in Bs and control group mosquitoes.

Results: Bs treatment remarkably reduced the infection rate of Plasmodium from 23.71% (124/523) to 16.23% (87/536) (Chi-square test, P =0.002 < 0.05), and the intensity of Plasmodium infection was greatly decreased in Bs group than that of the control group, either (Mann-Whitney Rank Sum Test, P< = 0.001). However, the rate of melanization mosquitoes was 4.97% (26/523) in the control group and 5.78% (31/536) in Bs group, respectively; no significant difference was detected between them (Chi-square test, P =0.558 > 0.05). Additionally, the intensities of melanized oocysts were compared between the two groups, and no significant difference was found, either (P =0.566>0.05). Interestingly, compared with the control group, the expression levels of TEP1 and Rel2 in Bs group were obviously up-regulated in larval, adult and infected mosquitoes. Especially in 3 dpi and 7 dpi, the expression level of TEP1 in Bs group was nearly 4 times higher than that of the control group, while Rel2 reached to approximately 7 times.

Conclusions: We firstly found that the sublethal dose of Bs significantly suppressed the vector competence of An. dirus to malaria parasites, which revealed a new important role of Bs on the basis of killing mosquito larvae. Furthermore, the Imd signaling pathway might play an effective way in Bs impacting the vector competence of An. dirus through upregulating the expression of NF-kB transcription factor Rel2, enhancing the expression of TEP1, which killed the Plasmodium, but not through melanization.

Keywords: Bacillus sphaericus; Anopheles dirus; Vector competence; Malaria; TEP1

Article history:

Received 14 September 2019

Received in revised form 26 September 2019

Accepted 30 September 2019

Available online 7 October 2019

 Identification and cross-species transmission of a novel ichthyic parvovirus in marine species in Hainan Island, China



Jiang Du1,2,3#, Wen-qi Wang1,2#, Jasper Fuk-Woo Chan1,4,5,6#, Gao-yu Wang1,2, Yi Huang1,2, Yu-fang Yi1,2, Zheng Zhu1,4,5,6#, Ruo-yan Peng1,2, Xiao-yuan Hu1,2, Yue Wu1,2, Ji-feng Zeng7,8, Ji-ping Zheng9, Xiu-ji Cui1,2,3, Li-na Niu1,2,3, Wei Zhao1,2,3, Gang Lu1,2,3, Kwok-Yung Yuen1,4,5,6, Fei-fei Yin1,2,3⊠

#These authors contributed equally to this work.

⊠Corresponding author: Fei-fei Yin, Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China.

E-mail: yinfeifeiff@163.com

Foundation project: This study was partly supported by the National Natural Science Foundation of China (Grant No. 81860367), State Key Laboratory of Virology 2018 Open Fund Project (2018IOV002).

1Hainan Medical University-The University of Hong Kong Joint Laboratory of Tropical Infectious Diseases, Hainan Medical University, Haikou, Hainan, China; and The University of Hong Kong, Pokfulam, Hong Kong

2Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, Hainan, China

3Department of Pathogen Biology, Hainan Medical University, Haikou, Hainan, China

4State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Pokfulam, Hong Kong

5Department of Microbiology, The University of Hong Kong, Pokfulam, Hong Kong

6Carol Yu Centre for Infection, The University of Hong Kong, Pokfulam, Hong Kong

7Key Laboratory of Tropical Animal Breeding and Epidemic Disease Research of Hainan Province, Hainan University, Haikou, Hainan, China

8Key Laboratory of Tropical Biological Resources of Ministry of Education, Haikou, Hainan, China

9Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Haikou, China

Parvoviruses are a diverse group of viruses that are capable of infecting a wide range of animals. In this study, we report the discovery of a novel parvovirus, tilapia parvovirus HMU-HKU, in the fecal samples of crocodiles and intestines of tilapia in Hainan province, China. The novel parvovirus was firstly identified from crocodiles fed with tilapia using next-generation sequencing. Screening studies revealed that the prevalence of the novel parvovirus in crocodile feces samples fed on tilapia (75%-86%) was apparently higher than that in crocodiles fed with chicken (4%). Further studies revealed that the prevalence of the novel parvovirus in tilapia feces samples collected at four areas in Hainan province was between 40% and 90%. Four stains of the novel parvovirusv were identified in this study based on sequence analyses of NS1 and all the four strains were found in tilapia in contrast only two of them were detected in crocodile feces. These findings proposed a possibility of cross-species transmission of the novel parvovirus from tilapia to crocodile. The nearly full-length genome sequence of the tilapia parvovirus HMU-HKU was determined and showed less than 28.3% and 25% amino acid identify with other members of Parvoviridae in NS1 and VP1 genes respectively. Phylogenetic analysis base on the NS1 and VP1 amino acid sequences showed that it was most closely related to members of the novel genus Chapparvovirus which did not belong to subfamily Parvovirinae nor Densovirinae. Tilapia parvovirus HMU-HKU formed an independent branch with Chapparvovirus and separated from it in the deep root of the phylogenetic tree. This indicated that the tilapia parvovirus HMU-HKU and Chapparvovirus viruses may had a common ancestor and they independently evolved in their natural hosts. The identification of tilapia parvovirus HMU-HKU provides further insight into the viral and genetic diversity of parvoviruses and its infections in tilapia populations need to be evaluated in terms of pathogenicity and production losses in tilapia farming.

Keywords: Parvovirus; Cross-species transmission; Crocodile, Tilapia; Hainan

Article history:

Received 19 September 2019

Revised 23 September 2019

Accepted 25 September 2019

Available online 7 October 2019