Paxillin和FAK mRNA在食管癌组织中的表达及临床意义

目的 研究桩蛋白(paxillin)和黏着斑激酶(focal adhesion kinase, FAK)mRNA在食管癌组织中的表达及其与临床病理学参数的关系,并分析两者在食管癌中表达的相关性.方法 运用RT-PCR技术检测59例食管癌组织、27例不典型增生组织和36例正常食管黏膜组织中paxillin和FAK mRN...     

2008-2010年哈尔滨市889例手足口病患者EV71病毒感染情况分析

目的了解2008-2010年哈尔滨市辖区医院收治的889例手足口病患者EV71病毒感染情况。方法采用两个样本率比较的假设检验,对889例手足口病患者咽拭子、粪便、肛拭子等标本RT-PCR或Real-time PCREV71核酸鉴定结果,按不同性别、不同居住地、不同年龄和不同发病时间分类进行比较,分析2008-2010年手足口病患者EV71感染情况。结果 3年合计,男性患者EV71阳性检出率高于女性(35.82%vs28.32%),农村患者阳性检出率最高(37.60%),城市(27.42%)与乡镇(27.36%)则相近;2~岁患者阳性检出率(37.56%)最高,其次为1~岁(34.98%)、3~岁(32.77%)和4~岁患者(32.69%),0~岁(29.63%)和5~岁(20.88%)相对较低;7月阳性检出率(38.11%)最高,其次为6月(35.48%)、5月(34.07%)。3年实验室监测确证EV71感染率分别为31.40%、25.00%和41.24%,3年间患者样本阳性检出率差异与分类情况有关。结论 2008-2010年哈尔滨市辖区医院收治的手足口病患者EV71阳性检出率按不同性别、不同居住地、不同年龄...     

应用荧光定量RT-PCR法检测牡蛎中诺如病毒

目的建立牡蛎样品中的诺如病毒检测方法。方法优化甘氨酸缓冲液处理牡蛎匀浆液,摸索PEG沉淀浓缩病毒的浓度,提取病毒RNA,采用荧光定量RT-PCR方法进行诺如病毒检测。应用优化后的方法检测105份牡蛎样本,并用核苷酸序列测定法对阳性PCR产物进行验证。结果甘氨酸缓冲液pH值为4.5,PEG沉淀浓度为16%时的诺如病毒富集提取效果最好,该方法的诺如病毒检出限为1.17×102个质粒拷贝。应用该方法对未知样本进行检测阳性率达14.29%,所得阳性PCR产物经核苷酸序列测序证实为诺如病毒。结论本研究方法可以应用于牡蛎中的诺如病毒检测,南宁市海鲜市场上的部分牡蛎含有诺如病毒。     

创伤后应激障碍大鼠海马组织5-HT1受体mRNA的表达

目的探讨创伤后应激障碍(posttraumatic stress disorder,PTSD)大鼠海马组织中5-羟色胺受体1mRNA的表达。方法随机将雄性Wistar大鼠分为对照组和PTSD组,采用改良的单一连续应激(SPS&S)方法刺激大鼠建立PTSD大鼠模型,Morris水迷宫实验检测大鼠记忆功能,化学发光法检测大鼠血清皮质醇浓度。建模后1d、4d、7d、14d分离海马组织,RT-PCR方法检测5-羟色胺受体1的变化。结果 (1)对照组水迷宫实验逃避潜伏期为(5.632±1.065)s,模型组为(20.762±3.236)s(t=9.932,P<0.01);(2)对照组血清皮质醇浓度为(1.25±0.12)μg/dl,模型组为(0.58±0.09)μg/dl(t=7.340,P<0.01);(3)应激后1d、4d、7d、14d海马5-HT1A受体mRNA表达相对水平分别为0.846±0.067、0.510±0.052、1.007±0.137、1.109±0.106,与对照组1.223±0.152相比较,1d、4d、7d表达低于对照组(P<0.05);5-羟色胺1受体其他亚型未见表达。结论 PTSD大鼠海马中5-HT1A受体mRNA表达降低。     

变应性哮喘患者外周血单个核细胞Tim-1 mRNA及Tim-3 mRNA表达及意义

目的研究Tim-1mRNA及Tim-3mRNA在变应性哮喘患者外周血单个核细胞(PBMC)的表达,探讨其在变应性哮喘发病中可能的作用。方法随机收集临床变应性哮喘患者25例,以健康体检者12例作为对照。采用实时荧光定量反转录-聚合酶链反应(RT-PCR)检测两组Tim-1mRNA和Tim-3的mRNA表达,并分析他们与哮喘临床表型之间的关系。结果 Tim-1mRNA和Tim-3mRNA在变应性哮喘患者PBMC中表达明显升高,差异有统计学意义(P<0.05),且与患者血清总IgE水平和临床严重程度相关,总IgE水平和临床严重程度分级越高,Tim-1mRNA和Tim-3mRNA表达越高(P<0.05)。结论 Tim-1mRNA和Tim-3mRNA在变应性哮喘患者中表达明显升高,Tim-1mRNA和Tim-3mRNA可能参与变应性哮喘的发生和发展。     

The role of NrCAM in neural development and disorders--beyond a simple glue in the brain

NrCAM is a neuronal cell adhesion molecule of the L1 family of immunoglobulin super family. It plays a wide variety of roles in neural development, including cell proliferation and differentiation, axon growth and guidance, synapse formation, and the formation of the myelinated nerve structure. NrCAM functions in cell adhesion and modulates signaling pathways in neural development through multiple molecular interactions with guidance and other factors. Alterations in NrCAM structure/expression are associated with psychiatric disorders such as autism and drug addiction and with tumor progression. The mechanisms of NrCAM participation in development and how these might be perturbed in disorders are reviewed.     

含CpG和CTB的HIV多表位基因真核表达载体的构建及表达

目的构建含非甲基化的CpG ODN序列及霍乱毒素B亚单位（CTB）的HIV复合多表位基因的真核表达载体,并在体外进行表达。方法设计并合成含CpG ODN序列和linker序列的CTB基因引物,用PCR方法扩增CpG-CTB基因（CC）,定向连接到真核表达载体pVL上,然后在CC基因下游插入HIV复合多表位基因MEGNp24,构建重组质粒pVL-CC-MEGNp24,酶切鉴定。纯化重组质粒,转染293T细胞,RT-PCR和细胞免疫染色方法分别检测CTB和p24基因的转录及表达,同时设质粒pVL及空白对照。结果构建了重组质粒pVL-CC-MEGNp24,该质粒转染293T细胞后,CTB和复合多表位基因在293T细胞中得到稳定表达。结论成功构建了含免疫佐剂的HIV复合多表位真核表达载体,为后期疫苗的研究奠定了基础。     

Surveillance of influenza A and the pandemic influenza A (H1N1) 2009 in sewage and surface water in the Netherlands

The role of the water cycle in spreading human pathogenic influenza viruses is poorly studied and is not considered to be significant. However, gastrointestinal symptoms developed in a large proportion of influenza A (H1N1) 2009 virus infected people during the pandemic in 2009 and fecal shedding was reported. This fecal route could potentially play a role in the entry of human pathogenic influenza viruses in to the water cycle. Monitoring of influenza viruses in sewage and surface water during the pandemic in 2009 showed that influenza A viruses were detected in sewage and surface water. However, the pandemic influenza A (H1N1) 2009 virus was not detected. These findings imply that the water cycle did not play a relevant role in spreading the pandemic influenza virus during the epidemic in the Netherlands in 2009. Analyses of deliberately contaminated water samples confirmed the ability of quantitative RT-PCR to detect influenza viruses in sewage samples whereas the analysis of large volumes of surface water was strongly hampered by the presence of PCR-inhibiting substances.     

乙肝血清标志物与HBV-DNA检测结果的相关性分析

目的:探讨乙肝血清标志物与HBV-DNA检测结果的相关性,以及二者联合检测在临床诊断中的应用。方法:收集1445例乙肝患者血清,同时采用电化学发光免疫分析法检测乙肝血清标志物和实时荧光定量PCR技术检HBV-DNA拷贝数。结果:HBsAg阳性HBeAg阳性组的HBV-DNA阳性率高于HBsAg阳性HBeAg阴性组和HBsAg阴性HBeAg阴性组(P<0.01),且HBsAg(+)、HBeAg(+)、抗HBc(+)类型中病毒载量集中在高拷贝区段,但在HBeAg阳性者中仍有部分HBV-DNA为阴性(10.9%),而在HBsAg阳性HBeAg阴性组中HBV-DNA阳性率仍有25.5%。结论:HBeAg阳性是反映病毒复制的良好指标,但无正相关;仅依靠乙肝血清标志物检测不能很准确地判断HBV复制的程度及其传染性的强弱;乙肝血清标志物与HBV-DNA检测的结合能更好地为临床提供准确可靠的诊断数据。