Expression of norovirus G Ⅱ.4 GZ121 strain P protein and analysis of its binding activity with HBGAs receptor / 中华微生物学和免疫学杂志

Objective To construct a prokaryotic expression system of norovirus (NoⅤ) G Ⅱ-4 strain P protein (P particle and P dimer) and to explore its binding activity and patterns with HBGAs receptor.Methods P domain sequence of GZ121 NoⅤ ORF2 gene was cloned and its phylogenic tree was constructed to identify the gene cluster.The pGEX-4T-1-based expression plasmids were constructed respectively by inserting P domain gene fragments with hinge and P-CDCRGDCFC without hinge,and then transformed into BL21 to express fusion proteins,which was induced with 0.6 mmol/L IPTG at 22℃ overnight.P proteins were purified by thrombin cutting and characterized by FPLC.The binding patterns of NoⅤ P protein to HBGAs antigens were analyzed by EIA.Results P region gene of GZ121 belonged to genotype G Ⅱ.4/2004 cluster.SDS-PAGE analysis showed the relative molecular weight of P particle and P dimer was about 36×103,which was consistent with other reports.The peak appeared at 830×103 confirmed the formation of P particle by FPLC.The expression of P protein was further confirmed by Western blot.The EIA results showed that GZ121 P protein could bind to saliva of A-group,B-group and O-group secretors,but not to nonsecretor.The binding affinity of P particle was 80-100 times higher than that of P dimer.Compared with VA387 P particle,it showed stronger binding affinity to O-group,but weaker to A-group.Conclusion The NoⅤ GⅡ-4 GZ121 P proteins including P particle and P dimer were successfully expressed and HBGAs receptor binding assays were established.This pave the way for further studies on the evolution dynamics of NoⅤ G Ⅱ.4 strains and the development of NoⅤ vaccines.

Experimental study on effect of baoganning on activity of nuclear transcription factor-kappaB in hepatic stellate cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Baoganning (BGN) on activity of nuclear transcription factor-kappaB (NF-kappaB) in hepatic stellate cells (HSCs) and its relevant mechanisms.</p><p><b>METHODS</b>Normal Wistar rats were medicated with BGN decoction by gavage for 7 days to prepare BGN drug-serum. The effect of BGN drug-serum on HSC-T6 growth was measured by MTT assay; phosphorylation level of NF-kappaB inhibiting factor IkappaB at different time after BGN stimulation was detected by Western blotting analysis; and the binding level of NF-kappaB with DNA was measured 30 min after drug-serum stimulation with gel shift assay.</p><p><b>RESULTS</b>BGN could significantly inhibit the HSC-T6 growth and quickly supress the phosphorylation of IkappaB, with the effect reached its peak at 30 min and restored to baseline level 6 h after stimulation, and reduce the binding capacity of NF-kappaB with DNA.</p><p><b>CONCLUSION</b>BGN can inhibit phosphorylation of IkappaB, restrain the activity of NF-kappaB and change the binding level of NF-kappaB with DNA.</p>

Progress on the targets of anti-noroviruses gastroenteritis agents / 中国药理学通报

Noroviruses are members of Caliciviridae family and are now the most commonly reported cause of outbreaks of nonbacterial gastroenteritis worldwide.Noroviruses gastroenteritis is difficult to control owing to the lack of a suitable antiviral agent or a vaccine to prevent infection.The tremendous disease burden of norovirus-associated gastroenteritis calls for an efficient preventive and therapeutic drugs.This review summarized the anti-noroviruses strategies targeting the nons-structural proteins,structural protein and so on.