Objective To clone and express the E2 gene in envelope region of hepatitis C virus and detect the anti E2 in sera of patients with HCV infection,using the purified E2 Protein.Methods The gene fragment of E2 region was obtained from the sera of patients with HCV infection by random primer reverse transcription and polymerase chain reaction(PCR),and then cloned into procaryotic expression system to express and Purify the protein and detect its antigenicity.Results A segment of E2 region gene with correct code reading frame was obtained,its full length was 984 bp and molecular weight of expressed Protein was 38kD.The detection of the protein performed by ELISA with sera from the patients with HCV infection,testified that it had appropriate antigenicity.Conclusion This experiment can provide the fundamental data for researching on the basic characteristics of HCV envelope protein,nuderstanding the significance of serological diagnosis and designing the anti HCV vassine.

Study on protein expression and antigenicity detection of hepatitis C Virus envelope protein E2 in prokaryotic and eucaryotic expression systems.The gene that encoding.HCV E2 protein was cloned in pQE30 and pEF1/HisC.After the expression of E2 protein in E.Coli M15 and COS 7 cell,the expressed proteins were used to detect their antigenicity with ELISA and WB.The results showed that protein E2 was expressed in both prokaryotic and eucaryotic cells.Special reaction could be detected using the expressed proteins and sera from HCV infected people.The studied E2 gene could express the desired proteins in both prokaryotic and eucaryotic expression systems,and glycosylation of the E2 protein happened in COS 7 cell.

Objective To investigate the dynamic change of pro‐and anti‐inflammatory eytokines of sepsis patients and its signif‐icance in clinical condition .Methods Forty‐three sepsis patients from 2010 to 2011 were divided into the survival group and the death group .Morning serum samples were collected on the first ,third ,firth and seventh day morning ;ELISA method was used to quantify the serum level of TNF‐α,IL‐1 ,IL‐4 and IL‐10 .The severity of patient′s condition was assessed according to the APACHEⅡsystem .Results In the early stage ,TNF‐α and IL‐1 in of both group increased and reached the peak on the third day ;then there was a gradual decline .Test in the same time point showed that the indexes of death group were all higher than that of survival group (P<0 .05) .IL‐4 of the two groups reached its peak on the fifth day and then declined ,and in the same time point ,indexes of death group were much more higher than that of survival group (P<0 .05) .IL‐10 of the survival group reached its peak on the fifth day and then declined;in the death group ,IL‐10 level kept increasing and maintained high ,there was no significant difference among the serum levels of the third ,fifth and seventh day(P>0 .05) .The APACHE Ⅱ of the survival group declined significantly while in death group it kept increasing and stay high .Conclusion Pro‐inflammatory eytokines(TNF‐α,IL‐1) ascended earlier than anti‐in‐flammatory eytokines(IL‐4 ,IL‐10) ,and the serum level of IL‐10 keep high level prompt the poor prognosis .