Tang Cuilan, Chen Zhi, Peng Guoping, Yang Zhenggang, Zhou Linfu
Key Laboratory of Health Ministry, Institute of Infectious Diseases, First Affiliated Hospital, School of Medicine, Zhejiang University, Zhejiang, China.
J Gastroenterol Hepatol. 2007 Jan;22(1):68-73. doi: 10.1111/j.1440-1746.2006.04371.x.
Chronic hepatitis B virus (HBV) infection remains a serious global health problem, inducing a spectrum of diseases, including asymptomatic HBV carriage (ASC) and chronic hepatitis B (CHB). ASC and CHB represent different immunological states and their prognoses are diverse. To clarify molecular mechanisms underlying the two infection states, the differentially expressed genes between the two states were screened and identified.
Subtracted complementary DNA libraries by suppression subtractive hybridization, dot blot hybridization and quantitative real-time PCR were used to identify the differentially expressed genes between subjects with CHB and those with ASC.
RNA from peripheral blood mononuclear cells from CHB and ASC subjects was subjected to suppression subtractive hybridization and resulted in isolation of subtracted complementary DNA clones. Eighty-eight randomly sampled clones were rescreened by dot blot hybridization, from which 29 clones were identified as differentially expressed genes. The differential expression of three genes was confirmed by real-time PCR in 23 subjects with CHB and 21 with ASC.
Differentially expressed genes in peripheral blood mononuclear cells between CHB and ASC have been isolated by suppression subtractive hybridization, including some new genes. Of the up-regulated genes in CHB, most are known to be responsive to inflammatory conditions. These genes might provide clues in elucidating the mechanisms of the two different HBV infection states and designing therapeutic targets for HBV infection.
慢性乙型肝炎病毒（HBV）感染仍然是一个严重的全球健康问题，可引发一系列疾病，包括无症状HBV携带（ASC）和慢性乙型肝炎（CHB）。ASC和CHB代表不同的免疫状态，其预后也各不相同。为阐明这两种感染状态背后的分子机制，对两种状态之间差异表达的基因进行了筛选和鉴定。
采用抑制性消减杂交构建消减互补DNA文库、斑点杂交和定量实时PCR来鉴定CHB患者和ASC患者之间差异表达的基因。
对CHB和ASC患者外周血单个核细胞的RNA进行抑制性消减杂交，分离出消减互补DNA克隆。通过斑点杂交对88个随机抽样的克隆进行重新筛选，从中鉴定出29个克隆为差异表达基因。通过实时PCR在23例CHB患者和21例ASC患者中证实了三个基因的差异表达。
通过抑制性消减杂交分离出了CHB和ASC患者外周血单个核细胞中差异表达的基因，包括一些新基因。在CHB中上调的基因中，大多数已知对炎症状态有反应。这些基因可能为阐明两种不同HBV感染状态的机制以及设计HBV感染的治疗靶点提供线索。
