小鼠脂多糖应答蛋白mlrpS的载体构建及融合蛋白表达

为构建小鼠mlrpS-cDNA基因原核、真核表达载体,大肠杆菌表达其融合蛋白。采用反转录-聚合酶链反应从经脂多糖刺激的鼠NIH3T3细胞cDNA中,扩增出编码mlrpS的cDNA。用限制性内切酶KpnⅠandXhoⅠ消化后,插入原核表达载体pTAT中,经酶切鉴定与测序证实后,转化大肠杆菌BL21(DE3)菌株。异丙基β-D硫代半乳糖苷(IPTG)诱导产生融合蛋白。经KpnⅠ和XhoⅠ酶切回收mlrpS-cDNA,插入pcDNA3.1载体中;pcDNA3.1-mlrpS再经KpnⅠ和ApaⅠ酶切后,插入到pEGFP-c1载体上,构建pEGFP-mlrpS融合的真核表达载体。构建mlrpS表达载体经测序证实,与GenBank登录的序列完全一致;双酶切鉴定证实,克隆基因正确插入载体pEGFP及pTAT;SDS-PAGE证实融合蛋白表达成功。说明:成功地构建了mlrpS原核、真核表达载体,成功正确表达了6His/mlrpS融合蛋白。

NLRP3炎症小体在心肌梗死发展的研究进展分析

心肌梗塞往往导致心肌坏死。对于心肌梗死的患者来说,冠脉血流及时再通是降低死亡率的关键,但再灌注可能对缺血的心肌造成二次损害,即心肌缺血再灌注损伤(MIRI)。慢性或越好奇不良左心室重塑是导致心肌梗死患者预后不良的重要因素。核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体通过级联放大炎症反应、促进动脉粥样硬化的发展和促进心肌细胞焦亡等,发挥其在心肌梗死整个过程中的重要作用,目前,关于NLRP3炎症小体及其相关抑制药物引起了临床广泛关注。有望在预防和治疗心肌梗死中提供新思路。

Research on Location-Routing Problem with Empirical Analysis for Regional Logistics Distribution

The location of the distribution facilities and the routing of the vehicles from these facilities are interdependent in many distribution systems. Such a concept recognizes the interdependence;attempts to integrate these two decisions have been limited. Multi-objective location-routing problem (MLRP) is combined with the facility location and the vehicle routing decision and satisfied the different objectives. Due to the problem complexity, simultaneous solution methods are limited, which are given in different objectives with conflicts in functions satisfied. Two kinds of optimal mathematical models are proposed for the solution of MLRP. Three methods have been emphatically developed for MLRP. MGA architecture makes it possible to search the solution space efficiently, which provides a path for searching the solution with two-objective LRP. At last the practical proof is given by random analysis for regional distribution with nine cities.