Influence of interferon-α therapy on CD8 T memory subsets in patients with chronic hepatitis B

Objective To investigate the influence of interferon-a therapy on CD8 T memory subsets in patients with chronic hepatitis B(CHB) and correlation between the effect of IFN-α and CD8 T memory subsets. Methods Blood samples from 57 patients with CHB were collected before treatment (0 week), at 12 weeks and 24 weeks of treatment with pegylated IFN-α. Assays were performed on freshly isolated peripheral blood mononuclear cells ( PBMCs). For phenotype analysis, All data were acquired on a flow cytometer instrument and prepared for analysis. Results A significantly higher frequency of CD8+ TEM and lower frequency of CD8+ TCM in inactive HBsAg carriers than that in CHB patients prior to treatment was observed (P <0.05). The proportion of CD8 + TCM was higher in group nonresponders than in group respond-ers, and the proportion of CD8 + TEM was lower in group nonresponders than in group responders (P < 0.05 ). The average dosage of IFN-α applied to patients with response was significantly higher than nonresponders. Conclusion The dominance of circulating effector memory T cells may be associated with elimination of viral infection, and possibly benefit for response to therapy with IFN-α.     

河南省和云南省HIV-1病毒env基因在脑脊液及血液中基因型分析

［摘要］　目的　了解河南省和云南省HIV-1病毒env基因在脑脊液及血液中的亚型变化。方法　收集6例河南省和5例云南省HIV-1阳性病人的血浆及脑脊液标本并提取RNA,逆转录为模版DNA后,采用巢式PCR法扩增env基因,对扩增产物进行核苷酸序列测定,构建系统进化树并进行离散度分析。结果　云南省各基因亚型与CRF07-BC、CRF08-BC和C亚型相近,河南省各基因亚型与B和B’亚型相近;河南人群脑脊液的测序结果显示患者1的部分克隆株与患者5的基因亚型一致,云南人群中12号患者和127号患者的血液及脑脊液的病毒亚型来自不同的分支,而其他患者的血液与脑脊液的病毒亚型一致。河南省HIV-1各亚型分离株脑脊液和血液间基因离散度差异有统计学意义(P<0.05)。结论　云南省HIV-1基因亚型为C亚型,而河南省为B和B’亚型。脑脊液和血液中的病毒亚型存在变异,中枢神经系统中HIIV病毒的离散度低于血液中HIIV病毒的离散度。     

Influence of interferon-α therapy on CD8 T memory subsets in patients with chronic hepatitis B

Objective To investigate the influence of interferon-a therapy on CD8 T memory subsets in patients with chronic hepatitis B(CHB) and correlation between the effect of IFN-α and CD8 T memory subsets. Methods Blood samples from 57 patients with CHB were collected before treatment (0 week), at 12 weeks and 24 weeks of treatment with pegylated IFN-α. Assays were performed on freshly isolated peripheral blood mononuclear cells ( PBMCs). For phenotype analysis, All data were acquired on a flow cytometer instrument and prepared for analysis. Results A significantly higher frequency of CD8+ TEM and lower frequency of CD8+ TCM in inactive HBsAg carriers than that in CHB patients prior to treatment was observed (P <0.05). The proportion of CD8 + TCM was higher in group nonresponders than in group respond-ers, and the proportion of CD8 + TEM was lower in group nonresponders than in group responders (P < 0.05 ). The average dosage of IFN-α applied to patients with response was significantly higher than nonresponders. Conclusion The dominance of circulating effector memory T cells may be associated with elimination of viral infection, and possibly benefit for response to therapy with IFN-α.     

非酒精性脂肪性肝炎大鼠肝脏内源性H_2S合成减少

目的探讨非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)大鼠肝脏硫化氢(hydrogen sulfide,H2S)变化及与肝细胞线粒体损伤的可能关系。方法采用雄性SD大鼠26只,分为正常对照组(n=6),NASH模型组(n=10)及复方牛胎肝提取物治疗组(n=10),高脂饮食12周建立NASH大鼠模型。亚甲基蓝分光光度法测定肝组织H2S生成率,RT-PCR及免疫组织化学测定肝组织CSEmRNA及其表达。透射电子显微镜观察肝细胞超微结构、PCR法测定肝细胞线粒体DNA(mtDNA)片段ND1、ND6、CO1、CYB及ATP6。结果 NASH大鼠模型组肝细胞线粒体肿大、膜模糊或嵴消失,正常组大鼠mtDNA片段ND1、ND6、CO1、CYB及ATP6表达明显高于模型组和治疗组(P<0.05)。NASH模型组肝组织H2S生成率〔(1.26±0.08)nmol/min〕较正常组〔(2.11±0.17)nmol/min〕明显减少(P<0.05),且肝脏CSEmRNA水平及其表达减少,治疗组与模型组比较差异无统计学意义。结论 NASH大鼠肝脏H2S合成减少可能与肝细胞线粒体损伤有关,H2S在NASH发病机制中可能具有保护肝脏的作用。     

DNA修复基因OGG1融合蛋白表达及多克隆抗体的制备

目的 表达及纯化8羟基脱氧鸟苷DNA糖苷酶-1(OGG1)融合蛋白制备多克隆抗体.方法 在大肠杆菌BL21中诱导表达OGG1融合蛋白,应用Ni-NTA亲和树脂进行纯化.纯化后的OGG1融合蛋白多次免疫大耳白兔,应用Westernb1otting的方法检测抗体活性,间接酶联免疫吸附分析(ELISA)法检测抗体效价.结果 ...     

泌尿生殖道解脲脲原体药敏403例分析

目的了解泌尿生殖道解脲脲原体(UU)对抗菌药物的敏感性,以指导临床合理用药。方法对本院门诊403例UU阳性患者的药敏结果进行回顾性分析。结果药敏试验中,美满霉素、克拉霉素、强力霉素及交沙霉素敏感率较高,依次为96.28%,95.53%,95.53%和90.82%,对氧氟沙星、司帕沙星、罗红霉素及阿奇霉素的耐药率较高,分别为19.85%,18.11%,12.66%和10.92%。结论治疗UU感染,临床应结合药敏试验合理用药,可以首选美满霉素、克拉霉素、强力霉素及交沙霉素。     

慢性HBV感染患者bcp/pc区点突变模式、pres区缺失及其临床意义

目的阐明HBV(hepatitis B virus,HBV)感染患者不同临床阶段bcp/pc(basic core promoter,BCP/precore,PC)点突变模式及pres区缺失突变规律,并探讨其临床意义。方法慢性HBV感染患者180例,其中,无症状HBV携带者13例,慢性乙型肝炎者75例,HBV相关肝硬化及肝癌分别为62、30例。Qiagen法提取血清HBV-DNA,常规PCR扩增目的基因,纯化PCR产物ABI377DNA自动测序仪直接双向测序。直接测序失败者,回收目的 DNA与PMD-18T载体连接,克隆质粒双向测序。DNAStar软件包的SeqMan软件进行生物信息分析。结果 Bcp/pc区点突变包括nt1753、nt1762、nt1764、nt1776、nt1803、nt1846、nt1896。HBV携带者、慢性乙型肝炎、HBV相关肝硬化及肝癌患者nt 1762(nt 1764)点突变率分别为7.7%、68.0%、72.7%(68.0%)及90.9%(81.8%),肝癌患者G1896A突变频率占54.6%。A1762T+G1764A联合突变占36%;A1762T、G1764A、G1896A联合突变占11%;T1753A/C、A1762T、G1764A、G1896A发生率为7%。克隆测序显示,肝硬化及肝癌患者bcp区起始点A1727G点突变率分别为72%、63%。HBeAg阴性患者存在更多的基因变异(P=0.022),G1776A和G1896A突变是HBeAg阴性的独立预测因素(P<0.05)。Bcp区点突变与HBeAg阴性无明显关系。肝硬化和肝癌患者pres基因缺失突变频率最高,肝癌及肝硬化患者pres1、pres2及pres1+s2缺失频率分别为7.1%、71.4%、7.1%及41.2%、58.8%、29.4%(P<0.05)。结论 A1727G、A1762T、G1764A及A1762T/G1764A联合突变、pres缺失在HBV相关肝硬化及肝癌患者多见,可能是肝脏疾病进展的危险因素,G1776A和G1896A突变是HBeAg阴性的独立预测因素。Bcp/pc点突变及pres区缺失可能为肝癌发生的早期预测因素,值得进一步研究。     

外周血CD4^＋ CD8^＋双阳性T细胞计数增加在肝脏疾病中的回顾性分析

[目的]通过分析CD4^＋ CD8^＋（DP）T细胞在肝脏疾病中的病因构成比，探讨DPT细胞在肝脏疾病中的意义。[方法]CD4^＋CD8^＋T细胞计数采用流式细胞术方法，使用BD公司Multiset软件测定，回顾性分析我院门诊及住院患者中DPT细胞。异常在肝脏疾病中的病因构成。[结果]在71例DPT细胞计数增加的肝脏疾病患者中，前4位的疾病分别是：乙型病毒性肝炎慢性重型、慢性活动性肝炎、肝移植术后和肝炎后肝硬化，分别占26．8％、12．7％、11．3％和9．9％。在感染性疾病和非感染性疾病中分别占70．4％和29．6％。[结论]DPT细胞计数增加在乙型病毒性肝炎慢性重型、慢性活动性肝炎中的比例较高，进一步研究DPT细胞计数增加的机制对于揭示病毒性肝炎的免疫发病机制具有重要意义。     

性病门诊就诊者泌尿生殖道性传播疾病的病原学检测分析

目的了解门诊就诊者泌尿生殖道病原体感染状况。方法收集北京佑安医院2005年7月～2006年5月门诊就诊者2 178例泌尿生殖道标本,进行病原体检测。结果2178份标本中有642份检出病原体,患病率为29.48%。单一感染450份(70.09%),其中检出淋球菌34份(29.82%),沙眼衣原体48份(44.44%),支原体363份(71.18%);混合感染192份(29.91%),其中淋球菌合并Ct或Uu者90份(24.75%),Ct Uu Mh102份(11.85%)。三种病原体在男女就诊者中的检出率有明显差异(P<0.01)。结论中青年就诊者的泌尿生殖道病原体阳性率较高,应重视病原学检查,进一步加强性病防治工作力度。