The Study of the mechanisms of fulminant hepatitis B using in vivo hepatitis model.

1993年，我们对一例重型B肝炎患者的HBV DNA进行了全序列分析，结果表明该病毒在包膜区、核心区、pol区和X区分别有4、12、15和6个错义突变。我们将突变的HBV DNA连接物转染到HepG 2细胞中，检测培养基中的病毒蛋白和肝癌细胞内的病毒复制。1995年，为了研究该突变病毒在体内的作用，我们建立了小鼠肝炎模型，在该模型中，病毒蛋白被分泌到循环中并在肝细胞中表达。我们将HBV DNA连接物和脂质体的混合物注射到小鼠肝被膜下。系列分析表明，HBsAg在循环中被检测到，然后在循环中从HBsAg血清转化为抗-HBs，并且HBsAg和HBcAg在小鼠肝细胞中表达。小鼠重型肝炎模型的建立研究正在进行中。

In 1993, We analyzed whole sequences of HBV DNA from an patient with fulminant hepatitis B.The result demonstrated that this viral DNA harbared 4,12,15 and 6 missense mutations in envelope, core, pol and X region, respectively.In 1994, in order to study the biological properties of the mutated HBV DNA from fulminant hepatitis B,we transfected the mutated HBV DNA constract into HepG2 cells and checked viral protein in culture medium and viral replication inside the hepatoma cells. As a result, this HBV DNA showed high level of production of HBV core protein and replicative intermediate.In 1995, in order to study how this mutant virus worked in vivo, we established murine hepatitis model, in which viral protein was secreted into circulation and expressed in hepatocytes. We injected the mixture of HBV DNA constract and lipofectin under the capsule of the murine liver. Serial analysis demonstrated that HBsAg was detected in circulation followed by the seroconversion from HBsAg to anti-HBs in circulation and HBsAg and HBcAg was expressed in murine hepatocyte. The study for the establishment of murine fulminant hepatitis model is ongoing.