肝内乙型肝炎病毒dna甲基化在核苷酸类似物停药复发中的作用分析word格式论文.docxVIP

肝内乙型肝炎病毒 DNA 甲基化在核苷（酸）类似物停药复发中的作用研究摘要 目的：研究核苷（酸）类似物治疗后达到停药标准的慢性乙型肝炎患者在停药时肝内乙型肝炎病毒 DNA 甲基化状态，从表观遗传学的角度探讨核苷类似物停药复发的分子病毒学机制。 方法：选择经核苷（酸）类似物治疗后达到停药标准（实现 HBeAg 血清转换、 HBVDNA 持续阴转后继续用药 24~48 周）的 HBeAg 阳性慢性乙型肝炎患者 18 例，进行肝组织活检，抽提肝内 HBV DNA，对 HBV DNA 进行亚硫酸氢盐化处 理，PCR 扩增 HBV 的 3 个 CpG 岛，克隆后测序。对所有患者每 4 周随访 1 次， 连续随访 24 周，观察停药后的病毒学应答情况，根据病毒学应答情况分为持久 应答组和停药复发组。分析比较停药复发组和持久应答组之间肝内 HBV DNA 甲 基化状态的差异。同时观察两组间停药时血清 HBsAg 定量水平的差异。 结果：停药时血清中 HBsAg 在停药复发组中的水平明显高于持久应答组，差异 具有统计学意义（p=0.037）。停药时肝内 HBV DNA 载量在两组间无显著性差异（p0.05）。停药时两组间肝内 HBV DNA 甲基化状态无统计学差异（p0.05）。 结论：核苷（酸）类似物停药时血清 HBsAg 水平越低越不易复发，可以作为停 药复发的预测指标。停药时肝内 HBV DNA 载量及 HBV DNA 甲基化状态能否作 为核苷（酸）类似物停药复发的预测指标需要进一步研究。 关键词：乙型肝炎病毒核苷（酸）类似物甲基化复发Study on the relationship between methylation of intrahepatic HBV DNA and viral relapse after nuclos(t)ide analogues withdrawnAbstractObjective: To analyze the methylation of intrahepatic HBV DNA in HBeAg positive chronic hepatitis B patients who received nuclos(t)ide analogues treatment and achived the cessation criterion, to investigate the molecular virological mechanism of .viral relapse after nuclos(t)ide analogues withdrawn.Methods: A total of 18 HBeAg positive chronic hepatitis B patients who received nuclos(t)ide analogues treatment and achived the cessation criterion (ie. HBeAg seroconversion and HBVDNA undetectable at least for 24~48 weeks) were enrolled in current study. The HBV DNA was extracted from the liver biopsy sample when antiviral agents withdrawn. Isolated DNA were modified with sodium bisulphite. The 3 targeted CpG island regions in HBV genome were amplified by PCR assay, then cloned and sequenced. All patients were followed up prospectively, once per 4 weeks, and then divieded into sustained response group and viral relapse group according to the viral load at week 24. The methylation patterns were analyzed in the two groups. Result:The level of hepatitis B virus surface antigen (HBsAg) at therapy cessation was significantly lower in the sustained response group than in th