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Background & objectives: Destruxin A, destruxin B and destruxin E isolated from entomopathogenic fungus Metarhizium anisopliae showed a strong suppressive effect on the replication of hepatitis B virus (HBV) in human hepatoma cells. In this study, the anti-HBV effects of the crude destruxins extracted from M. anisopliae var. dcjhyium were detected both in vitro and in vivo. Methods: HepG2.2.15 cells were cultured to observe the inhibitory effects of the crude destruxins on the gene expression and replication of HBV by radioimmunoassay detection and real-time quantitative PCR. In vivo, duck HBV (DHBV)-infected ducks were treated with the crude destruxins at 2.0, 4.0, 6.0 μg/kg once a day for 15 days, DHBV DNA was examined by real-time quantitative PCR. Results: The crude destruxins suppressed the replication of HBV-DNA and the production of HBsAg and HBeAg with IC50 of about 1.2 and 1.4 μg/ml. Transcript of viral mRNA was significantly suppressed by the crude destruxins in HepG2.2.15 cells. In vivo, the duck serum DHBV-DNA levels were markedly reduced in the group of the crude destruxins. Interpretation & conclusions: The crude destruxins inhibited the gene expression and replication of HBV both in vitro and in vivo, and their anti-HBV effect was stronger than that with destruxin B. Our results indicate that the crude destruxins from M.anisopliae var. dcjhyium may be potential antivirus agents. Further studies need to be done to confirm these findings
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OBJECTIVE: To study the inhibitory effects of traditional chinese medicine Compound Liuyuexue (CLYX) on hepatitis B surface antigen(DHBsAg) and hepatitis B e-antigen(DHBeAg).METHODS: One-day old guangxi brown spotted ducks infected with DHBV were used as the hepatitis B virus infected animal model. Positive ducks were detected by PCR at 13 days after the infection of DHBV, and were randomly divided into five groups: the high dose group, middle dose group and low dose group of Compound Liuyuexue(CLYX), model group, and positive control group, with 10 ducks in each group. CLYX was given ig.for 14 days. Serum sampling was scheduled at 0, 7, 14 days respectively and 3 days after drug withdrawal, and the contents of DHBsAg and DHBeAg in serum were measured by ELISA. RESULTS: The serum DHBsAg and DHBeAg contents in high dose and middle dose groups of CLYX were decreased significantly(P
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Objective IFN-? is a pleiotropic cytokine with potent immunomodulatory effects and antiviral activity. To study the mechanism of IFN-? clearing duck hepatitis B virus (DHBV) in ducks, it is essential to establish a method to quantify expression of DuIFN-? in immune response. In the present study,a semi-quantitative competitive RT-PCR was developed to quantify expression of duck IFN-?(DuIFN-?) mRNA by PBMCs. Methods Based on ?-actin consensus sequence, fishing the ?-actin gene as house keeping gene from duck PBMC by RT-PCR. A competitive internal control was constructed and the competitive RT-PCR system could be used to quantify the transcription of DuIFN-? mRNA. Results After duck PBMCs were stimulated in vitro with PHA, the peak of DuIFN-? expression was at 24-36h. Then RT-PCR method was applied to detect DuIFN-? mRNA transcription by PBMCs from DHBV infected ducks immunized with DuIFN-? plasmid plus DNA vaccine or DNA vaccine alone. Results showed that expression of DuIFN-? in ducks co-immunized with DuIFN-? plasmid were higher than other groups immunized without DuIFN-? plasmid as adjuvant. Conclusions The results indicated that DuIFN-? gene could be a useful adjuvant to develop vaccines. The semi-quantitation of DuIFN-? mRNA by competitive RT-PCR provides the basis for future study of the mechanism of IFN-? in duck hepatitis B virus persistent infection.
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Objective:To study the specific photochemical effects of a newly designed target photosensitizer. Methods Based on the technique of antisense nucleic acid and the principle of photochemical reaction effects,a specific sensitizer,TFO P has been designed and synthesized.When in coordination with long wave ultraviolet ray(UVA) ,this decorated complex (TFO P) was added into the blood cell suspension to inactivate the contaminating virus( duck hepatitis virus B,DHBV).The efficacy of specific binding to DHBV DNA and viral inactivation by TFO P was detected by gel shift blot assay and infection of primary culture of duck hepatocyte.Results The designed TFO P could specifically bind to different DHBV DNA line sample and present different linking level.With a TFO P concentration of 0.1 nmol/ml and UVA intensity of 1800 ?W/cm 2,the DHBV in blood cell suspension could be reduced by 1.90～5.40 logs.Conclcusion The photochemical effects of TFO P could significant inactivate DHBV in blood.