The anti-hepatitis B virus (HBV) effects and its mechanisms of the ethanol extracts of Hypericum perforatum L. (EHP) in vitro were explored. HepG2 2.2.15 cells, a stable HBV-producing cell line, were cultured as the model system to observe the anti-HBV effect. The viral antigens of cellular secretion, HBsAg and HBeAg, were determined by enzyme linked immunosorbent assay (ELISA). The quantity of HBV-DNA released in the supernatant was assayed by real-time PCR. In order to understand the mechanisms of the suppression of HBV replication, all HBV promoters (Cp, Xp, S1p, S2p and Fp) with luciferase reporter gene were transfected into HepG2 cells respectively. Then the activities of viral promoters were examined by luciferase reporter assay. It was found EHP effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose-dependent manner, as well as the extracellular HBV DNA. And EHP could selectively inhibit the activity of HBV promoter Fp. Our data suggest that EHP exerts anti-HBV effects via inhibition of HBV transcription, which helps to elucidate the mechanism underlying the potential therapeutic value of EHP.

In this study, the anti-HBV effects of tea polyphenols (TP) were examined. After cells were exposed to TP for 3, 6, 9 days, amounts of HBsAg, HBeAg and HBV-DNA released into the supernatant of the cultured HepG2 2.2.15 cells were detected. TP, to some extent, inhibited the secretion of HBsAg and strongly suppressed the secretion of HBeAg in a dose-dependent (P<0.01) and time-dependent manner, with 50% maximal inhibitory concentration (IC50) value being 7.34 microg/mL on the 9th day, but the time-dependence was not significant (P=0.051). Expression of HBV-DNA in the supernatant of the cell culture also was significantly decreased in a dose-dependent fashion (P<0.01). The IC50 of TP in inhibiting HBV DNA was 2.54 microg/mL. It concluded that TP possessed potential anti-HBV effects and may be used as a treatment alternative for HBV infection.
Antiviral Agents/*pharmacology ; DNA, Viral/analysis ; Dose-Response Relationship, Drug ; Flavonoids/*pharmacology ; Hep G2 Cells ; Hepatitis B Surface Antigens/analysis ; Hepatitis B e Antigens/analysis ; Hepatitis B virus/*drug effects ; Inhibitory Concentration 50 ; Phenols/*pharmacology ; Tea/*chemistry

OBJECTIVETo establish a mouse model for human chronic HBV infection, and to investigate the role of PD-1/PD-L1 signaling pathway in antiviral immunity.

METHODSA mouse model was established by hydrodynamic injection of the plasmid pAAV/HBV1.2-GFP into the tail vein of C57BL/6 mice, HBV markers were assayed at different time points after injection. After intraperitoneal injection of anti-PD-L1 monoclonal antibody, the serum ALT, and HBV DNA in the serum, liver and kidney were assayed.

RESULTSThe chronic HBV infection mouse model were established successfully, serum HBsAg and high load of HBV DNA were detectable 90 days after plasmid injection. After blocking of the PD-1/PD-L1 pathway, the serum ALT level of mice were significantly increased (P < 0.01), and the HBV DNA load in serum (P < 0.01), liver (P < 0.05) and kidney (P < 0.05) were decreased significantly.

CONCLUSIONBlocking the PD-1/PD-L1 signaling pathway can enhance antiviral response in mice with chronic HBV infection.

Animals ; Antigens, Surface ; metabolism ; Apoptosis Regulatory Proteins ; metabolism ; B7-1 Antigen ; metabolism ; B7-H1 Antigen ; DNA, Viral ; analysis ; Disease Models, Animal ; Hepatitis B virus ; metabolism ; Hepatitis B, Chronic ; immunology ; metabolism ; virology ; Male ; Membrane Glycoproteins ; metabolism ; Mice ; Mice, Inbred C57BL ; Peptides ; metabolism ; Programmed Cell Death 1 Receptor ; Signal Transduction

Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Apoptosis Regulatory Proteins ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; blood ; Humans ; Middle Aged ; Programmed Cell Death 1 Receptor ; Up-Regulation ; Young Adult

In this study,the anti-HBV effects of tea polyphenols (TP) were examined.After cells were exposed to TP for 3,6,9 days,amounts of HBsAg,HBeAg and HBV-DNA released into the supernatant of the cultured HepG2 2.2.15 cells were detected.TP,to some extent,inhibited the secre-tion of HBsAg and strongly suppressed the secretion of HBeAg in a dose-dependent (P<0.01) and time-dependent manner,with 50% maximal inhibitory concentration (IC50) value being 7.34 μg/mL on the 9th day,but the time-dependence was not significant (P=0.051).Expression of HBV-DNA in the supernatant of the cell culture also was significantly decreased in a dose-dependent fashion (P<0.01).The IC50 of TP in inhibiting HBV DNA was 2.54 μg/mL.It concluded that TP possessed potential anti-HBV effects and may be used as a treatment alternative for HBV infection.

The anti-hepatitis B virus(HBV)effects and its mechanisms of the ethanol extracts of Hypericum perforatum L.(EHP)in vitro were explored.HepG2 2.2.15 cells,a stable HBV-producing cell line,were cultured as the model system to observe the anti-HBV effect.The viral antigens of cellular secretion,HBsAg and HBeAg,were determined by enzyme linked immunosorbent assay(ELISA).The quantity of HBV-DNA released in the supernatant was assayed by real-time PCR.In order to understand the mechanisms of the suppression of HBV replication,all HBV promoters(Cp,Xp,S1p,S2p and Fp)with luciferase reporter gene were transfected into HepG2 cells respectively.Then the activities of viral promoters were examined by luciferase reporter assay.It was found EHP effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose-dependent manner,as well as the extracellular HBV DNA.And EHP could selectively inhibit the activity of HBV promoter Fp.Our data suggest that EHP exerts anti-HBV effects via inhibition of HBV transcription,which helps to elucidate the mechanism underlying the potential therapeutic value of EHP.

Objective:To explore the effect of early clinical clerkship training in the early exposure to clinical practice of eight-year program medical students.Methods:Experimental control method was used in this study. A total of 120 eight-year program medical students in the third year of Batch 2014 to 2016 from the Union Hospital, Tongji Medical College, Huazhong University of Science and Technology were selected as the research objects. The students of each grade were randomly selected by computer and divided into experimental group and control group, with 20 students in each group. Before early clinical exposure, the experimental group received 16 class hours of early clinical clerkship training, while the control group did not receive early clinical clerkship training. After the early clinical exposure, the clinical diagnosis and treatment ability of the two groups of students was compared. SPSS 24.0 software was used for t test. Results:The scores of medical history inquiry of experimental group and control group were [(17.45±1.96) points and (15.95±1.93) points; (18.30±1.03) points and (16.75±1.86) points; (17.95±1.36) points and (16.40±1.60) points, respectively]. The physical examination scores were [(17.75±1.65) points and (16.05±1.64) points; (17.85±1.18) points and (16.80±1.47) points; (18.25±1.16) points and (16.85±1.63) points, respectively]. The clinical judgment scores were [(18.15±1.42) points and (16.35±2.41) points; (18.20±1.24) points and (16.65±1.53) points; (18.35±1.35) points and (16.25±1.83) points, respectively]. Diagnosis and treatment scheme scores were [(17.15±1.57) points and (14.55±2.56) points; (17.30±1.42) points and (15.90±1.48) points; (17.80±1.06) points and (16.35±1.87) points, respectively]. The scores of communication skills were [(17.95±1.15) points and (17.00±1.19) points; (18.55±0.83) points and (17.45±1.50) points; (18.45±1.00) points and (17.45±1.23) points, respectively], with statistically significant differences (all P<0.05). Conclusion:The application of early clinical clerkship training in the early exposure to clinical practice of eight-year program medical students can improve the quality of students' clerkship.