四川地区不同疾病谱慢性感染者病毒基因分型与_四川建筑杂志社投稿

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四川地区不同疾病谱慢性感染者病毒基因分型与

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Abstract:ObjectiveTo investigate the genotype distribution of chronic hepatitis B virus (HBV) infection patients with different disease spectrums in Sichuan, China, as well as the association of genotypes with cellular immunity and coagulation total of 543 patients with chronic HBV infection who were hospitalized in Chengdu Public Health Medical Center from January 2015 to June 2017 were enrolled, among whom there were 31 chronic HBV carriers, 285 patients with chronic hepatitis B (CHB), 77 patients with severe hepatitis B, 81 patients with hepatitis B cirrhosis, and 69 patients with hepatocellular carcinoma (HCC). Blood samples were collected; gene microarray was used to determine the genotypes of HBV DNA, and T lymphocyte subsets and coagulation function were measured. An analysis of variance was used for comparison of continuous data between multiple groups, and the SNK-qtest was used for further comparison between two groups; the chi-square test was used for comparison of categorical data between the 543 patients with chronic HBV infection, 400 (73.66%) had type B infection, 123 (22.65%) had type C infection, 10 (1.84%) had mixed type B and C infection, and 10 (1.84%) had mixed type C and D infection. There were no significant differences in the distribution of HBV genotypes, prothrombin time, and prothrombin activity in different disease spectrums between the patients with type B infection and those with type C infection (allP>0.05). According to the results of T lymphocyte subsets, in the severe hepatitis B group, the patients with type C infection had higher percentages of CD3+and CD3+CD4+T cells than those with type B infection; in the chronic hepatitis B group, the patients with type C infection had a higher percentage of CD3+CD8+T cells than those with type B infection; in the chronic HBV carrier group, severe hepatitis B group, and hepatitis B cirrhosis group, the patients with type C infection had a higher CD4+/CD8+ratio than those with type B infection; in the hepatitis B cirrhosis group, the patients with type C infection had lower percentages of CD3+, CD3+CD4+, and CD3+CD8+T cells than those with type B infection; in the HCC group, the patients with type C infection had lower percentages of CD3+, CD3+CD4+, and CD3+CD8+T cells and a lower CD4+/CD8+ratio than those with type B infection, with significant differences in the percentages of CD3+CD4+T cells (t=2.078,P=0.041) and CD3+CD8+T cells (t=6.672,P<0.001) between the patients with type C infection and those with type B infection in the hepatitis B cirrhosis B and C are the main genotypes of chronic HBV infection in different disease spectrums in Sichuan, with type B more commonly seen than type C, and there is no difference in the distribution of HBV genotypes between patients with different disease spectrums. Disorder of cellular immune function is observed in patients with chronic HBV infection, with different immune disorders in different stages of infection, and the genotype of infection is not associated with coagulation function and cellular immune function.

Keywords：hepatitis B virus; genotype; immunity, cellular; blood coagulation; Sichuan

HBV感染机体后主要通过宿主免疫机制引起肝损伤，造成HBV感染后不同临床转归的原因有病毒、环境因素、宿主遗传因素等[1]，不同的病毒基因型可能影响抗原表达而产生不同的临床表现，具有不同的流行特征及致病性。肝脏在凝血机制中占有极其重要的地位，凝血因子合成能力反映了肝细胞的功能状态。国外学者Sass等[2]认为凝血时间是判断重型肝炎预后的重要因素。不同疾病谱慢性HBV感染者基因型是否导致机体免疫功能的差异及凝血功能的改变，临床报道不多。本研究通过对四川地区慢性HBV携带者、慢性乙型肝炎、重症乙型肝炎、乙型肝炎肝硬化及肝细胞癌患者的基因型和T淋巴细胞及凝血功能变化进行分析，以探讨不同基因型与细胞免疫及凝血功能的关系。

1 资料与方法

1.1 研究对象选取2015年1月-2017年6月于本院住院的四川地区(宜宾、巴中、广安、广元、成都、甘孜州、阿坝州)HBV感染者。根据患者症状、体征、实验室及影像学检查结果，将患者分为慢性HBV携带者、慢性乙型肝炎、重症乙型肝炎、乙型肝炎肝硬化以及肝细胞癌(在慢性乙型肝炎基础上发生的肝细胞癌)5组，诊断符合《慢性乙型肝炎防治指南(2015年更新版)》[3]。纳入患者均进行HBV DNA基因分型以及T淋巴细胞亚群和凝血功能检测。排除甲型肝炎、丙型肝炎、戊型肝炎、HIV、酒精性肝病、自身免疫性肝炎、药物性、代谢性等疾病。患者均签署知情同意书。

1.2 HBV DNA基因分型检测采用基因芯片法，使用珠海赛乐奇生物技术有限公司提供的仪器及配套试剂盒，将待测不同HBV基因型的特征基因片段(靶基因)固定于玻片上制成检测芯片，从患者血清中抽提出的HBV DNA经PCR扩增标记后与芯片进行杂交，杂交信号由扫描仪扫描，再经计算机分析确认结果。严格按照试剂盒上说明进行操作。