Aim:Regulated endothelial exocytosis of Weibel-Palade bodies(WPBs),the first stage in leukocyte traffickin9,plays a pivotal role in vascular inflammation and *** hypertensive mechanical stress has been closely associa...
详细信息
Aim:Regulated endothelial exocytosis of Weibel-Palade bodies(WPBs),the first stage in leukocyte traffickin9,plays a pivotal role in vascular inflammation and *** hypertensive mechanical stress has been closely associated with inflammation although the precise mechanism is unknown.
目的:本研究改进基于脱氧核糖核酸(DNA)连接酶的中低通量基因分型方法,并用此方法进行核苷酸结合寡聚结构域(NOD)样受体基因NOD1和NOD2与冠心病的关联分析。方法:通过多重聚合酶链式反应(PCR)预扩增增加连接模板分子数量,提高特异性连接效率,优化DNA探针设计,摸索连接反应温度、时间、循环圈数及连接酶种类实现等位基因特异性连接,通过荧光掺入PCR和毛细管电泳实现多种等位基因特异性产物一次性检测。Sanger测序验证此方法准确性后,利用此方法对NOD1和NOD2基因上的单核苷酸多态性(SNP)位点在1 555例冠心病患者和1887例对照受试者中进行分型和关联分析。结果:通过优化反应条件和等位基因特异性探针设计原则,实现10 ng DNA样本一次性分型30个等位基因多态性位点。基于改进的DNA连接酶中低通量基因分型方法,NOD1、NOD2基因与冠心病的关联分析发现,NOD2基因上rs1861759和rs751271位点在非高血压条件下与冠心病相关(P均<0.05),经Bonferroni多重检验校正后,相关性仍然显著(P均<0.05)。结论:本研究优化了DNA连接酶链式反应技术在设计上的关键点,联合使用多重PCR和毛细管电泳,建立了一种高准确性、低成本、满足基于中低通量位点分型的临床分子诊断和科研需求的基因分型新方法,并用该方法发现NOD2基因上的位点rs1861759和rs751271在非高血压条件下与冠心病相关。
Regulated endothelial exocytosis of Weibel-Palade bodies(WPBs),the first stage in leukocyte trafficking,plays a pivotal role in vascular inflammation and *** mechanical stress has been closely associated with inflamma...
详细信息
Regulated endothelial exocytosis of Weibel-Palade bodies(WPBs),the first stage in leukocyte trafficking,plays a pivotal role in vascular inflammation and *** mechanical stress has been closely associated with inflammation although the precise mechanism is *** we show that mechanical stretch regulates the exocytosis of
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