乙型肝炎肝硬化与HLA-DRB1*1301,1302等位基因的相关性研究

目的 探讨人类白细胞抗原(HLA)的DRB1*1301,1302基因与乙型肝炎肝硬化的关系.方法 采用序列特异性引物聚合酶链反应(PCR-SSP)对27例乙型肝炎肝硬化和30例慢性乙型肝炎患者进行HLA-DRB1*1301,1302的检测.结果 HLA-DRB1*1301,1302在乙型肝炎肝硬化组高于慢性乙型肝炎组(29.6%vs10.0%,χ2=5.900,P<0.05,RR=5.294),差异有统计学意义.结论 HLA-DRB1*1301,1302基因与慢性乙型肝炎发展成肝硬化可能有关.     

慢乙肝患者急性发作后血清HBV DNA的动态变化

将慢性乙型肝炎（慢乙肝）患者分为观察组（急性发作期）和对照组,观察组应用甘利欣等治疗,对照组未予以治疗。分别于入院后次日和第31 d应用常规赖氏法检测血清ALT、比色法检测AST、聚合酶链反应检测HBV DNA。结果观察组治疗前ALT、AST和HBV DNA均高于对照组,治疗后均低于治疗前（P〈0.01,〈0.05）;ALT、AST与HBV DNA无直接相关关系（P〉0.05）。提示慢乙肝急性发作后,随着转氨酶升高,HBV DNA降低,乙肝病毒开始被清除。     

Expression profile of altered long non-coding RNAs in patients with HBV-associated hepatocellular carcinoma

In China, hepatitis B virus (HBV) infection is the major cause of hepatocellular carcinoma (HCC), which is called HBV-related HCC (HBV-HCC), but the pathogenesis has not been clearly elucidated. Long non-coding RNAs (lncRNAs) have been paid increasing attention to, as an important regulatory molecule involved in many biological processes, as well as a variety of diseases. This study examined lncRNA that might play an important role in HBV-HCC pathogenesis by conducting lncRNA and mRNA profile comparison between HBV-HCC and normal liver tissues. The differentially expressed lncRNA and mRNA profiles between HBV-HCC and normal liver tissues were analyzed by microarrays. The potential protein-encoding gene regulated by lncRNA, and the biological function (gene ontology, pathway analysis) of the target gene were investigated. The results showed that the expression levels of lncRNA and mRNA in HBV-HCC tissues were different from those in normal liver tissues. As compared with normal liver tissue, 837 (4.30%) lncRNAs exhibited more than two-fold change (P<0.05); 325 were up-regulated, and 512 were down-regulated; 991 (5.70%) mRNAs exhibited more than 2-fold change (P<0.05); 733 were up-regulated and 258 were down-regulated in HBV-HCC tissue. Besides, there were 7 lncRNAs with above 10-fold elevation, 6 lncRNAs with above 10-fold decrease, 18 mRNAs with above 10-fold elevation and 11 mRNAs with above 10-fold decrease. 444 (53.05%) lncRNAs had their corresponding mRNAs, some of which were adjacent to lncRNAs. The biological analysis showed that the target gene of differentially expressed lncRNAs took part in the important bio- logical regulatory function. Target gene-related pathway analysis revealed the pathways in carcinoma and mitogen-activated protein kinase (MAPK) signaling pathways significantly changed in the HBV-HCC tissues as compared with normal liver tissues (P<0.05). It was suggested that as compared with normal liver tissues, the expression of lncRNAs in HBV-HCC tissues changed significantly, and lncRNAs played a key role in the pathogenesis of HBV-HCC probably by mainly regulating the carcinoma-related signaling pathway and MAPK signaling pathways.     

慢性乙型肝炎患者拉米夫定耐药模式及耐药后体内免疫环境的改变

目的探讨慢性乙型肝炎患者发生拉米夫定耐药常见的模式及耐药后体内免疫环境的改变。方法应用PCR及基因测序方法回顾性分析72例发生拉米夫定耐药患者血清HBVDNA多聚酶区基因变异模式,比较72例患者拉米夫定抗病毒6个月时及耐药后血清INF-γ、IL-2、IL-4因子水平与对照组细胞因子的水平差异。结果拉米夫定耐药4种主要的突变模式分别为：单位点rtM204I突变,占34.7%（25/72）;rtL180M＋rtM204V,占30.56%（22/72）;rtL180M＋rtM204I,占16.67%（12/72）;rtV173L＋rtV180M＋rtM204V,占12.5%（9/72）;抗病毒6个月时,未变异组与对照组IL-2（P=0.20）、IFN-γ（P=0.14）、IL-4（P=0.119）水平差异无统计学意义。耐药组HBVDNA发生病毒变异后,IL-2、IFN-γ水平显著低于对照组（P〈0.01）,而IL-4水平则高于对照组（P〈0.01）,差异有统计学意义。结论慢性乙型肝炎应用拉米夫定抗病毒治疗耐药模式主要是rtM204I突变及rtL180M、rtV173L、rtM204V、rtM204I联合变异,发生耐药后慢性乙型肝炎患者体内免疫环境将从Th1优势应答转换为Th2优势应答。     

头孢地嗪对小鼠T细胞亚群的影响

目的研究头孢地嗪对小鼠的免疫调节作用。方法对小鼠分别连续7d注射胸腺肽10mg/kg、头孢地嗪(500mg/kg、200mg/kg、50mg/kg)、头孢曲松500mg/kg及生理盐水后,经流式细胞仪对小鼠T细胞亚群进行检测。结果胸腺肽组、头孢地嗪(500mg/kg、200mg/kg、50mg/kg)组CD4 /CD8 均明显高于头孢曲松组和生理盐水组(P<0.05),而头孢曲松组与生理盐水组无差异(P>0.05)。结论头孢地嗪可通过调整CD4 和CD8 之间的平衡,有效地提高机体的免疫功能。     

四种影像学检查评价原发性肝癌患者介入治疗的疗效

目的 探讨超声造影、螺旋CT、MRI在原发性肝癌（PLC）经导管肝动脉化疗栓塞术（TACE）治疗后近期疗效评估的价值及意义,为临床在PLC介入治疗术后复查方式选择提供经验依据。方法 选择2010年6月至2015年6月在郑州大学第一附属医院就诊的临床资料及影像学资料完整的PLC行TACE治疗患者45例,术后复查行数字减影血管造影（DSA）、超声造影、螺旋CT和MRI检查,以DSA为金标准,对比分析三种影像学检查对TACE治疗后肿瘤残留及新发病灶检出的灵敏度Se）和特异度（Sp）。结果 在小于5 cm病灶的诊断中,MRI（Se=92.3％,Sp=88.5％）和超声造影（Se=89.4％,Sp=84.6％）的灵敏度及特异度显著高于螺旋CT（Se=88.7％,Sp=80.8％）,差异具有统计学意义（P＜0.05）,在大于5 cm的病灶诊断中,三种影像学检查的效能无统计学差异（P＞0.05）;在诊断病灶供血动脉来源方面,三种影像学检查的效能差异无显著性统计学差异（P＞0.05）;在诊断门静脉癌栓方面,螺旋CT的准确性（92.2％）显著高于MRI（90.8％）或超声造影（90.5％）,差异具有统计学意义（P＜0.05）;在观察病灶强化预示复发方面,超声造影（95％）、MRI（96.4％）的特异度显著优于螺旋CT（93.7％）,且其差异具有统计学意义（P＜0.05）。结论 超声造影检查可作为螺旋CT和MRI之外评估PLC患者介入治疗后肿瘤残留及新发病灶检出的另一有效影像学检查,具有较大的临床应用价值。     

全程监测加联合用药对乙型肝炎疗效分析

目的探索合理而有效的慢性乙型肝炙长期治疗方案,分析全程督导加联合用药对慢性乙型肝炎治疗效果、方法对在1996年～2002年间二次治疗的678例慢性乙型肝炙患者,进行抗病毒或不进行抗病毒治疗,两次检测患者血清Ⅳ-C和PCⅢ,多因素回归统计学分析结果678例患者进行多因素回归分析表明,抗病毒治疗时间在所有因素中影响最大;其次为监督治疗时间,紧接着是抗纤维化治疗时间,最后分别是非特异性免疫治疗时间和其它治疗方案时间、人为分组后,全程督导治疗加联合治疗的治疗组治疗效果较好,血清学检测指标与其它组相比,有统计学差异、结论在全程监督治疗的情况下,联合抗病毒治疗(干扰素),免疫调节(胸腺肽)和抗纤维化的联合治疗为最佳治疗方案,适宜于慢性乙型肝炎长期抗纤维化治疗。     

慢性乙型肝炎急性发作的免疫学机制

目的　通过检测慢性乙型肝炎患者急性发作后体液免疫和细胞免疫的相关指标,探讨慢性乙型肝炎急性发作后宿主免疫功能变化及其与肝功能损伤之间的关系,为阐明慢性乙型肝炎的发病机制和特异性免疫治疗提供理论基础。方法　25例随机入组的慢性乙型肝炎患者,根据 HBc-IgM定性结果分为两组(HBc-IgM阳性组为实验组;HBc-IgM阴性组为对照组),分别以赖氏法检测血清丙氨酸氨基转移酶(alanine aminotransferaseALT)、比色法检测天冬氨酸氨基转移酶(aspartate aminotransferase AST)、核酸扩增(PCR)荧光定量检测乙型肝炎病毒(HBV) DNA、激光散射比浊法测定血清免疫球蛋白G(IgG)和荧光素双标记 FCM(flow cytometry)技术检测外周血CD3 、CD4 、CD8 T淋巴细胞相对含量,统计学分析用 t检验与秩和检验。结果　HBc-IgM阳性组ALT、AST和HBVDNA平均值高于 HBc-IgM阴性组,有统计学差异(P<0.05);两组血清 IgG含量和外周血CD3 、CD4 、CD8 T细胞亚群间无统计学差异(P>0.05)。结论　慢性乙型肝炎急性发作后,HBV清除延迟;HBV激活的宿主免疫应答主要通过特异性免疫导致肝组织损伤和病毒清除,宿主体内多克隆免疫应答的激活不明显。