Objective: To investigate the inhibitory effects against dengue virus serotype 2 (DENV-2) by five different fractions (extracted by methanol, ethanol, benzene, chloroform and n-hexane) of Rumex dentatus, Commelina benghalensis, Ajuga bracteosa and Ziziphus mauritiana, as well as their constituents (gallic acid, emodin, and isovanillic acid). Methods: All the samples were tested for cytotoxicity on baby hamster kidney cells by MTT assay and for anti-DENV-2 activity by plaque reduction neutralization assay using two DENV-2 doses (45 and 90 plaque- forming units or PFU). Results: All the samples except isovanillic acid exhibited significant prophylactic effects against DENV-2 infectivity (without cytotoxicity) when administered to cells before infection, but were not effective when given 6 h post-infection. The methanol extract of Rumex dentatus demonstrated the highest antiviral efficacy by inhibiting DENV-2 replication, with IC₅₀ of 0.154 μg/mL and 0.234 μg/mL, when added before infection with 45 and 90 PFU of virus, respectively. Gallic acid also exhibited significant antiviral effects by prophylactic treatment prior to virus adsorption on cells, with IC₅₀ of 0.191 μg/mL and 0.522μg/ mL at 45 and 90 PFU of DENV-2 infection, respectively. Conclusions: The highly potent activities of the extracts and constituent compounds of these plants against DENV-2 infectivity highlight their potential as targets for further research to identify novel antiviral agents against dengue.

Objective: To investigate the effect of Cannabis sativa extract on the development of neuro- and hepato-toxicity caused by malathion injection in rats. Methods: The extract of Cannabis sativa was obtained from the plant resin by chloroform treatment. Δ-Tetrahydrocannabinol content of the extract (20%) was quantified using gas chromatography–mass spectrometry. The doses of cannabis extract were expressed as Δ -tetrahydrocannabinol content of 10 or 20 mg/kg. Malathion (150 mg/kg) was intraperitoneally administered followed after 30 min by the cannabis extract (10 or 20 mg/kg, subcutaneously). Rats were euthanized 4 h later. Malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide and paraoxonase-1 (PON-1) activity were determined in brain and liver. Brain 5-lipoxygenase and butyrylcholinesterase (BChE) activity were measured as well. Histopathological examination of brain and liver tissue was also performed. Results: Compared to controls, malathion resulted in increased oxidative stress in brain and liver. MDA and nitric oxide concentrations were significantly increased (P<0.05) and GSH significantly decreased with respect to control levels (P<0.05). Malathion also significantly inhibited PON-1 and BChE activities but had no effect on brain 5-lipoxygenase. Brain MDA concentrations were not altered by cannabis treatment. Cannabis at 20 mg/kg, however, caused significant increase in nitric oxide and restored the GSH and PON-1 activity. Brain BChE activity significantly decreased by 26.1% (P<0.05) after treatment with 10 mg/kg cannabis. Cannabis showed no effect on brain 5-lipoxygenase. On the other hand, rats treated with cannabis exhibited significantly higher levels of liver MDA, nitric oxide and PON-1 activity compared with the malathion control group. Rats treated with only malathion exhibited spongiform changes, neuronal damage in the cerebral cortex and degeneration of some Purkinje cells in the cerebellum. There were also hepatic vacuolar degeneration and dilated and congested portal vein. These histopthological changes induced by malathion in brain and liver were reduced to great extent by cannabis administration at 20 mg/kg. Conclusions: Our data suggest that acute treatment with cannabis alleviates the malathion-induced brain and hepatic injury in rats possibly by maintaining the levels of GSH and PON-1 activity.

Objective: To investigate the phytochemical constituents, antioxidant and anthelmintic activities of the crude methanol extract of Cissus populnea (C. populnea) rhizomes. Methods: Phytochemical screening was performed using standard protocols, and column chromatography of silica gel was used for the compounds isolation. DPPH antiradical scavenging assay was performed in order to evaluate the antioxidant activity. Total phenolic content was evaluated using the Folin–Ciocalteu assay. The anthelmintic activity was screened on the bovine adult male forms of parasitic nematode Onchocerca ochengi, by the in vitro evaluation of the inhibition of adult worm motility and mortality. Worms were incubated in the presence of different concentrations of the plant extract and effects on survival were monitored after 24 and 48 h. Results: The preliminary phytochemical screening revealed the presence of phenolic compounds, saponins, steroids, tannins, and terpenoids. Bergenin and a mixture of phytosterol, β -sitosterol and stigmasterol were isolated from this extract and were identified by nuclear magnetic resonance, mass spectrometry and by comparison with published data. The crude methanol extract of C. populnea rhizomes showed a strong DPPH antiradical activity with a good amount of total phenolic content ((20.69±2.13) g gallic acid equivalent/100 g of extract) and significant anthelmintic activity comparable to the standard drug ivermectin. Bergenin was found to be inactive even after 72 h of incubation. Conclusions: This study constitutes the first report on the anthelmintic activity of this plant and supports the traditional use of C. populnea as a natural antioxidant and anthelmintic.

Objective: To investigate the antioxidant and α-glucosidase properties and phytochemical constituents of roots, stems, leaves and flowers extracts and aerial parts oil of Chrysanthoglossum trifurcatum (Desf.) (C. trifurcatum). Methods: For extraction from roots, stems, leaves and flowers of C. trifurcatum, methanol, ethyl acetate and petroleum ether solvents were used. Phenols, flavonoids, flavonols and tannins contents were evaluated. More, C. trifurcatum aerial parts oil composition was determined using chromatography/mass spectrometry. The antioxidant effect was estimated by DPPH, ABTS and reducing power test systems. The α-glucosidase inhibition was determined by colorimetric assay using the enzyme from Aspergillus niger and the p-nitrophenyl glucopyranoside (pNPG) as substrate. Results: The highest amounts of polyphenols, flavonoids, flavonols and tannins were shown by the methanolic extract of leaves. The main components of the aerial parts oil were limonene (29.21%), γ -terpinene (12.96%), 4-terpenyl acetate (12.18%) and α -pinene (5.76%). The activity evaluated by DPPH, ABTS and reducing power tests was important for stems (IC₅₀=0.68 mg/mL) and flowers (IC₅₀=0.67 mg/mL) methanolic extracts and essential oil (IC₅₀=0.72 mg/mL). Findings of α-glucosidase activity revealed that petroleum ether extracts of leaves and roots together with aerial parts oil showed a highest activity with IC₅₀ of 0.044, 0.045 and 0.049 mg/mL, respectively. Conclusions: Observed antioxidant and α-glucosidase activities of oil and extracts are attributed to the presence of the active phytochemicals in C. trifurcatum organs. Thus, the C. trifurcatum can be used as a source of antioxidant compounds and dietary supplement to treat patients with type 2 diabetes.

Objective: To discover the mechanism behind ameliorative effects of Michelia champaca (M. champaca) in gastrointestinal, respiratory and cardiovascular disorders. Methods: Anti- spasmodic potential was evaluated by trying the M. champaca extract (aqueous:ethanolic) on rabbit aorta, trachea and jejunum in vitro. Isotonic and isometric transducers coupled with Power Lab data acquisition system was used to record the responses of isolated tissues. Results: M. champaca extract relaxed the spontaneous and high K⁺ (80 mmol/L)-induced contractions of isolated jejunum preparation of rabbit showing a Ca²⁺ channel blocking mechanism. Moreover, extract shifted calcium concentration response curves towards right like standard calcium channel blocker verapamil. In rabbit tracheal preparation, M. champaca relaxed both carbachol (1 1 μmol/L) and high K⁺-induced contractions, likewise verapamil. In rabbit aorta preparation, M. champaca relaxed phenylephrine (1 μmol/L) and high K⁺-induced contractions similar to verapamil. Conclusion: M. champaca possesses spasmolytic, airways relaxant and vasodilator actions mediated perhaps due to blocking of Ca²⁺ channels, hence validating its therapeutic usage in diarrhea, asthma and hypertension.

Objective: To explore the expression level and prognostic significance of erythropoietin receptor (EPOR) in patients with breast cancer (BRCA) based on estrogen receptor (ER) status and different molecular subtypes. Methods: The Cancer Genome Atlas (TCGA) and GTEx data were collected in GEPIA initially to identify the dysregulated genes. Further, bc- GenExMiner 4.1 online bioinformatics tool was used to evaluate EPOR mRNA differential expression level according to different classification of clinicopathologic parameters in patients with breast cancer. Additionally, the prognostic value between EPOR mRNA expression and free survival of metastatic relapse (MR) or any event (AE, namely any relapse or death) in patients with breast cancer was done. Results: EPOR mRNA was significantly downregulated in BRCA (1 085 cases) compared to normal tissues (291 cases) (P<0.05). Univariate Cox analysis revealed that high EPOR mRNA expression was remarkably correlated to a decreased risk of MR (HR: 0.79, P<0.000 1) and AE (HR: 0.87, P =0.000 7) especially in breast cancer patients with ER+. Besides, high EPOR mRNA level also associated with a favorable MR- free survival (HR: 0.81, P =0.007 2) and AE-free survival (HR: 0.88, P =0.029 9) in ER+ breast cancer patients. However, no similar above phenomenon was detected in ER- patients. Moreover, the subsequent prognostic adjusted analyses and univariate Cox analysis of AE based on SSP or SCM molecular subtypes validated the above results. Conclusion: EPOR mRNA level is a valuable prognostic indicator for patients with ER+ breast cancer.

Objective: To investigate the risk factors of chronic obstructive pulmonary disease (COPD) in Li and Han ethnic group in Hainan, China. Methods: All subjects were randomly selected from various regions in Hainan. General characteristics were compared between COPD cases and healthy control cases in both Li and Han ethnic groups. The odds ratio (OR), the corresponding 95% confidence interval (CI) of COPD were calculated by logistic regression. Results: A total of 277 Li COPD cases, 307 Li healthy control subjects, 290 Han COPD cases and 301 Han healthy control were included in this study. In both the Li and Han groups, the average age exceeded 65 years, and the cigarette number smoked per day and the smoking duration were correlated with risk of COPD. In the Li COPD subjects, low weight, smoking, and recurrent infection of respiratory tract were mainly risk factors; while the mainly risk factor of Han COPD subjects was family history of respiratory disease. Conclusions: The risk factors are different in COPD subjects of Han and Li nationalities in Hainan of China. The age and smoking are strongly correlated with COPD risk.

Objective: To investigate the relationship between polymorphisms of rs1532624 and rs289741 loci in cholesteryl ester transfer protein (CETP) genes and atherosclerotic cerebral infarction (ACI). Methods: The CETP gene rs1532624 and rs289741 in 95 patients with ACI and 177 healthy subjects were genotyped by MassARRAY mass spectrometry. Each locus genotype and allele frequency distributions were compared. Results: The difference of allele frequency distribution between the rs1532624 (χ²=1.723, P =0.189) and rs289741 (χ²=2.466, P =0.116) were not statistically significant. The frequency distribution of rs1532624 genotype between the cerebral infarction group and healthy control group was statistically significant (χ²=7.096, P =0.029), while rs289741 genotype frequency distribution between the two groups was not statistically significant (χ=2.906, P =0.234). Conclusion: ACI have a positive correlation with rs1532624 polymorphism, and AA genotype may be susceptible factors of ACI.