购买
下载掌阅APP,畅读海量书库
立即打开
畅读海量书库
扫码下载掌阅APP

第二章
乙型肝炎重症化的研究方法和技术

内容提要

1.乙型肝炎重症化的实验研究常用技术和方法包括乙型肝炎病毒的检测(含病毒基因变异的检测)、乙型肝炎病毒与细胞相互作用的检测(凝胶迁移率变动分析、染色质免疫沉淀法、免疫共沉淀、酵母双杂交系统、细胞凋亡的检测等)、免疫细胞功能的检测(HBV抗原特异性细胞毒性T淋巴细胞的检测、非特异性细胞的功能检测)、免疫学技术(流式细胞术、固相酶联免疫斑点技术、四聚体/五聚体技术等)、遗传学及表观遗传学技术(甲基化特异性检测、荧光原位杂交技术等)、肠道微生态学研究的技术(光冈氏肠内菌群分析方法等)等。

2.现有的乙型肝炎病毒细胞模型包括原代肝细胞感染模型、稳定转染肝癌细胞系模型和替代细胞模型,它们在研究HBV发病机制、抗病毒药物的筛选中发挥了一定的作用。

3.目前乙型肝炎的动物模型有了较大研究进展,但都有其局限性。急性肝衰竭动物模型的建立主要有病毒诱导法、化学药物诱导法、外科手术诱导法和基因敲除诱导法等,其中以MHV-3感染近交系小鼠诱导的暴发性肝衰竭模型为病毒诱导法的代表。

4.近年来,由于针对乙型肝炎重症化病因的治疗(抗病毒治疗)受到人们重视,不少旨在探讨乙型肝炎重症化内科治疗手段的临床研究已开展。乙型肝炎重症化干预性临床研究设计的重点问题包括病例选择、对照的考虑、样本量、研究终点选择及疗效判断标准、安全性评价、混杂因素的考虑、统计学要求、伦理学要求和综合考虑新药的药学、药理毒理特点等。设计合理的试验方案并严格执行是乙型肝炎重症化干预性临床研究成功的关键。

Abstract 2

1.Major research methods and technologies in patients with severehepatitis B include detection of HBV(including detection of virus genetic variations),the interaction between HBV and cells(e.g., by gel mobility fluctuation analysis,chromatin immuno-precipitation,co-immuno precipitation,yeast two-hybrid system,and assays of cell apoptosis),the function of cells of the immune system(e.g.,detection of HBV antigen specific CTLs and testing of nonspecific cell function),and experimental immunological technique(e.g.,flow cytometry,solid phase enzyme linked immunospot technology,and four/five polymer technology),genetics,epigenetics (e.g.,methylation and fluorescence in situhybridization)and intestinal microecology (e.g.,analysis of intestinal flora).

2.Current cell models of HBV infection include those involving primaryhepatocytes,stable transfected liver cancer cells and alternative cells.These models play important roles in studying the pathogenesis of HBV and for screening antiviral drugs,but eachhas some limitations.

3.Therehas been great progress in the use of animal models ofhepatitis B,but these models alsohave limitations.Animal models of acute liver failurehave been established virologically,chemically,surgically and by gene knockout.One representative model is MHV-3-induced acute liver failure in BALB/cJ mice.

4.In recent years,therehas been considerable clinical research investigating the medical treatment of severehepatitis B.The key to clinical research design includes selection of cases and controls,sample size,end points and efficacy measures,safety evaluation,confounding factors,statistics,ethics,pharmacology and toxicology.A reasonable research plan and its strict implement are keys to success. 90Cx71gXQ/QSpjbNu7JkVzY07zwpRsenQaAWZVB3DkfWUGZHjByd3tO+OGjK/F0r

点击中间区域
呼出菜单
上一章
目录
下一章
×