背景:血管内皮发生的分子机制是细胞及基因替代治疗的首要前提。目的:构建一种通过血管内皮细胞特异蛋白TIE2启动子来启动表达的E1A激活基因阻遏子(cellular repressor of E1A-stimulated genes,CREG)和增强绿色荧光蛋白(enhanced green fluorescence protein,EGFP)融合的真核表达质粒。方法:根据GenBank中公布的TIE2基因的启动子序列,人工合成TIE2启动子DNA序列,经AseⅠ和NheⅠ双酶切后,亚克隆入表达质粒pEGFP-N1中构建pTIE2-EGFP-N1。同时,用BamHⅠ和EcoRⅠ双酶切pcDNA3.1myc-His/hCREG质粒得到CREG基因,亚克隆入质粒pTIE2-EGFP-N1中构建pTIE2-CREG-EGFP-N1,酶切鉴定。应用脂质体法将该质粒转染至体外培养的小鼠动脉内皮细胞,荧光显微镜下观察EGFP的表达;Western blot检测CREG蛋白的表达。结果与结论:经酶切鉴定证实实验构建的pTIE2-CREG-EGFP-N1质粒正确;体外转染48h,荧光显微镜下可见EGFP的表达,Western blot检测到CREG蛋白的表达。说明实验成功构建了pTIE2-CREG-EGFP-N1重组质粒,其可携带目的基因在小鼠动脉内皮细胞中有效表达。
Background Some larger scale, randomized studies have demonstrated the superiority of drug-eluting stents （DES） over bare metal stents （BMS） for the treatment of acute myocardial infarction （AMI）. This study aimed to investigate the impact of DES, in comparison with BMS, on the 2-year clinical outcomes in patients with ST-elevation myocardial infarction （STEMI）.Methods From January 2002 to December 2008, a total of 1301 consecutive STEMI patients treated with coronary stenting in Shenyang Northern Hospital were prospectively registered. Patients received BMS （n=868） or DES （n=435） implantation in the infarction related artery according to physician＇s discretion. A propensity score analysis was performed and two well matched subgroups were selected （BMS, n=288; DES, n=288） to evaluate the 2-year clinical outcomes. The primary outcome was the occurrence of major adverse cardiac events （MACE）, which was defined as a composite of all-cause death, myocardial infarction （MI）, or target vessel revascularization （TVR）.Results Survival salvage analysis showed that 2-year cumulative hazards were not significantly different between the two groups with respect to TVR （2.8% vs. 3.1%, log-rank P=0.780）, stent thrombosis （1.7% vs. 4.2%, log-rank P=0.079） and MACE （8% vs. 10.8%, log-rank P=0.236）. Multivariate analysis showed that DES was an independent protective factor of MI （HR： 0.211, 95% CI： 0.049 to 0.908） and stent thrombosis （HR： 0.327, 95% CI： 0.107 to 0.994）.Conclusion DES was associated with similar 2-year clinical outcomes to those of BMS for the treatment of STEMI in daily practice.
LI YiHAN Ya-lingZHANG Quan-yuGUAN Shao-yiWANG Xiao-zengJING Quan-minMA Ying-yanWANG GengWANG BinDENG Jie
