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心血管疾病基因治疗的基础研究Ⅱ.Pro-UK和t-pA基因的体外表达
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作者 姚阿卿 孙双丹 +6 位作者 李岱宗 温进坤 王贵春 周爱儒 黄培堂 俞炜源 汤健 《北京大学学报(医学版)》 CAS CSCD 1994年第S1期115-119,共5页
构建了带有人的全长Pro-UK或t-pA基因cDNA的逆转录病毒质粒pN2-CMV-UK或pN2-CMV-tPA。重组质粒转染包装细胞PA3l7后获得含假病毒的培养上清。用假病毒上清感染培养的血管平滑肌细胞(VsMC... 构建了带有人的全长Pro-UK或t-pA基因cDNA的逆转录病毒质粒pN2-CMV-UK或pN2-CMV-tPA。重组质粒转染包装细胞PA3l7后获得含假病毒的培养上清。用假病毒上清感染培养的血管平滑肌细胞(VsMC),经G4l8筛选得到抗性集落。Southernblot分析表明,Pro-UK或t-pA基因已整合到宿主细胞染色体。在转染的VSMC中,Pro-UK和t-pA的含量分别为265ng/107细胞/24小时和5.6ng/107细胞/24小时。溶圈法测定结果证明,Pro-UK基因导入细胞后可以表达出有生物活性的尿激酶原。 展开更多
关键词 pro-ukt-pa基因 基因治疗 血管平滑肌细胞
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Development of nano t-PA gene coating stent and its preventive effect on dog coronary artery stent thrombosis
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作者 ZHAO Hong-lei HU wei +3 位作者 CHEN Qi-ying ZENG Fan-fang LONG Juan WANG Li-li 《South China Journal of Cardiology》 CAS 2017年第3期223-230,共8页
Background The regulation of t-PA gene is the essence and core of thrombosis. Therefore, the present study aimed to prepare nano t-PA gene coated stent and to observe its effect on coronary stent thrombosis in dogs.Me... Background The regulation of t-PA gene is the essence and core of thrombosis. Therefore, the present study aimed to prepare nano t-PA gene coated stent and to observe its effect on coronary stent thrombosis in dogs.Methods Highly expressed t-PA gene plasmid was constructed and albumin nano t-PA gene coating stent was prepared. The major branch vessels of dog coronary artery were pre-expanded with a 3.0 mm×20 balloon with 8-10 atmospheric pressure. 10 dogs of the control group were implanted with bare metal stent; while 12 dogs of the experimental group were implanted with nano t-PA gene coating stent. Both groups were not given anti-coagulation treatments. Blood samples were taken for t-PA and D-dimer before the operation, at 1,2,4 and 8 weeks after operation. Pathological analysis found thrombosis in the cavity and the hyperplasia of the intima. t-PA expression was detected by immunohistochemical indirectly, and the thickness of the intima of the section cross was directly measured by morphometry. Liver, heart, kidneys and lung were taken for pathological observation. Results All experimental animals survived at postoperative 8 weeks. Vascular stent thrombosis was seen in 10 cases of the control group with the thrombosis rate of 100%; while was seen in 2 cases among 12 cases of the experimental group with the thrombosis rate was 16.67%(P=0.00087). Immunohistochemical staining showed that the positive t-PA gene transfection of the experimental group was mainly distributed on the surface of hyperplasia intima, and vascular wall t-PA expression of the control group was negative. Positive t-PA signal was not found in the liver,heart, kidneys and lung. Conclusion Nano t-PA gene vector coating stent can effectively express t-PA in vascular wall and effectively prevents stent thrombosis. 展开更多
关键词 STENTING THROMBOSIS nano t-pa gene vectors coating stent
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