在大量前期工作基础上，计划利用来自临床DHF病人的DEN-2临床分离株和国际参考株，构建 EDⅠ结构域原核表达质粒和表达蛋白，分别感染或作用于原代和传代人VEC，系统地研究DEN导致VECs凋亡的机制；观察其细胞因子产生的规律；细胞通透性的改变和超微结构的变化；并通过基因芯片实验、数据分析、real-time qRT-PCR、配对t检验分析感染组与未感染组间筛选差异基因的基因本体(Gene Ontology，GO)；分析差异基因主要参与的通路，分析VECs被DEN感染后基因水平的变化；通过体内FITC-白蛋白通透性试验 白蛋白可通过损伤的血脑屏障进入脑实质，可通过检测脑组织中荧光强度来反映血脑屏障的的通透性增加。应用FITC-白蛋白为示踪剂, 监测血脑屏障的损伤程度，为DHF／DSS的发病机制的阐明提供充分的科学依据。

A large number of pre-work basis, plans to use from the clinical DHF patients with DEN-2 clinical separation and the international reference strains, and build the ED Ⅰ, domain of the Prokaryotic expression plasmid and protein expression, respectively, infection or role in the primary and transfer on behalf of people VEC, the system reseach of DEN led to the mechanism of apoptosis of the VECs; observe the laws of their cytokine production; cell permeability changes and ultrastructural changes; microarray experiments, data analysis, real-time qRT-PCR, and paired t-testscreening of differentially expressed genes, gene ontology analysis of the infected group and uninfected group (gene Ontology, GO); analysis of differentially expressed genes involved in the pathway analysis the VECs DEN infection levels of gene changes; by in vivo FITC-albumin permeability testalbumin may damage the blood-brain barrier into the brain parenchyma, to reflect the increased permeability of the blood-brain barrier of the fluorescence intensity in the detection of brain tissue. With FITC-albumin as a tracer to monitor the degree of damage of the blood-brain barrier to provide adequate scientific basis for the elucidation of the pathogenesis of DHF / DSS.