The P19CL6 mouse embryonic carcinoma cells efficiently differentiate into cardiac muscle cells in the presence of DMSO. A reporter plasmid for cardiac muscle differentiation was constructed by connecting the CMV enhan...The P19CL6 mouse embryonic carcinoma cells efficiently differentiate into cardiac muscle cells in the presence of DMSO. A reporter plasmid for cardiac muscle differentiation was constructed by connecting the CMV enhancer and a 250 bp MLC-2v promoter in front of the GFP gene to further evaluate the role of the CMV enhancer. This plasmid (pCBVenh/MLC-2vpro/EGFP) was stably introduced into P19CL6 cells, and the transfectant differentiated into cardiomyocytes with DMSO. Upon DMSO addition, GFP was immediately transcribed (within 2 days) and the amount of the transcript increased with cultivation. Concomitantly, GFP fluorescence was detected in the cells under a microscope. However, native MLC-2v was transcribed later on day 4. This expression time course is different from that of GFP. Clearly the CMV enhancer responded immediately to DMSO. Since GATA DNA-binding proteins play crucial roles in the initiation of cardiomyocyte differentiation, such a response could be ascribed to the presence of multiple GATA motifs in the enhancer sequence but not in the native MLC-2v promoter. Thus the CMV enhancer may be not only useful for gene therapy and monitoring cell differentiation but also the study of the role of GATA transcription factors expressed in P19CL6 cells.展开更多
Our previous studies demonstrated that CD151 gene promoted neovascularization in ischemic heart model.To improve the delivery efficacy and target specificity of CD151 gene to ischemic heart,we generated an adeno-assoc...Our previous studies demonstrated that CD151 gene promoted neovascularization in ischemic heart model.To improve the delivery efficacy and target specificity of CD151 gene to ischemic heart,we generated an adeno-associated virus（AAV） vector in which CD151 expression was controlled by the myosin light chain（MLC-2v） promoter to achieve the cardiac-specific expression of CD151 gene in ischemic myocardium and to limit unwanted CD151 expression in extracardiac organs.The function of this vector was examined in rat ischemic myocardium model.The protein expression of CD151 in the ischemic myocardium areas,liver and kidney was confirmed by using Western blot,while the microvessels within ischemic myocardium areas were detected by using immunohistochemistry.The results showed that MLC-2v significantly enhanced the expression of CD151 in ischemic myocardium,but attenuated its expression in other organs.The forced CD151 expression could increase the number of microvessels in the ischemic myocardium.This study demonstrates the AAV-mediated and MLC-2v regulated CD151 gene is highly expressed in the ischemic myocardium and cardiac-specific delivery that is more efficiently targets CD151 to the ischemia myocardium after myocardial infarction.展开更多
目的对微光刀立体定向放射治疗系统(Dual Camera 2 mm Stereotactic Radiosurgerg,D2SRS)临床剂量学可靠性进行测试、研究。方法随机选取20例已接受放射治疗的病例,其中包括7例颅内肿瘤、6例肺部肿瘤和7例肝癌病例,将20例病例的CT图像和...目的对微光刀立体定向放射治疗系统(Dual Camera 2 mm Stereotactic Radiosurgerg,D2SRS)临床剂量学可靠性进行测试、研究。方法随机选取20例已接受放射治疗的病例,其中包括7例颅内肿瘤、6例肺部肿瘤和7例肝癌病例,将20例病例的CT图像和RT Structures通过DICOM网络分别传输至雷泰Ti GRT计划系统和Eclipse11.0计划系统,进行放射治疗计划设计,统计两组靶区及危及器官的剂量学参数并进行统计学分析,将两组调强计划分别进行绝对剂量和二维面剂量验证,并比较两组计划的绝对剂量和二维平面剂量验证结果。结果两组计划均能满足临床剂量要求,靶区剂量学参数D_(98)、D_(50)、D_(02)、CI以及HI均无统计学差异(P>0.05);危及器官受照剂量如全脑、心脏、食管、肝脏和肾脏的D_(mean)、脑干和脊髓D_(max)、双肺V_(20)等剂量学参数均无统计学差异(P>0.05);Eclipse与Ti GRT两组计划的验证结果显示,点剂量误差统计结果为(0.70±1.44,0.57±1.86,t=0.268,P=0.796);二维面剂量Gamma分析统计结果为(96.8%±0.9%,97.3%±1.2%,t=2.031,P=0.056)。结论D2SRS在剂量精确性上达到同类型进口设备相似水平。展开更多
文摘The P19CL6 mouse embryonic carcinoma cells efficiently differentiate into cardiac muscle cells in the presence of DMSO. A reporter plasmid for cardiac muscle differentiation was constructed by connecting the CMV enhancer and a 250 bp MLC-2v promoter in front of the GFP gene to further evaluate the role of the CMV enhancer. This plasmid (pCBVenh/MLC-2vpro/EGFP) was stably introduced into P19CL6 cells, and the transfectant differentiated into cardiomyocytes with DMSO. Upon DMSO addition, GFP was immediately transcribed (within 2 days) and the amount of the transcript increased with cultivation. Concomitantly, GFP fluorescence was detected in the cells under a microscope. However, native MLC-2v was transcribed later on day 4. This expression time course is different from that of GFP. Clearly the CMV enhancer responded immediately to DMSO. Since GATA DNA-binding proteins play crucial roles in the initiation of cardiomyocyte differentiation, such a response could be ascribed to the presence of multiple GATA motifs in the enhancer sequence but not in the native MLC-2v promoter. Thus the CMV enhancer may be not only useful for gene therapy and monitoring cell differentiation but also the study of the role of GATA transcription factors expressed in P19CL6 cells.
基金supported by a grant from the National Natural Sciences Foundation of China (No. 30670856)
文摘Our previous studies demonstrated that CD151 gene promoted neovascularization in ischemic heart model.To improve the delivery efficacy and target specificity of CD151 gene to ischemic heart,we generated an adeno-associated virus（AAV） vector in which CD151 expression was controlled by the myosin light chain（MLC-2v） promoter to achieve the cardiac-specific expression of CD151 gene in ischemic myocardium and to limit unwanted CD151 expression in extracardiac organs.The function of this vector was examined in rat ischemic myocardium model.The protein expression of CD151 in the ischemic myocardium areas,liver and kidney was confirmed by using Western blot,while the microvessels within ischemic myocardium areas were detected by using immunohistochemistry.The results showed that MLC-2v significantly enhanced the expression of CD151 in ischemic myocardium,but attenuated its expression in other organs.The forced CD151 expression could increase the number of microvessels in the ischemic myocardium.This study demonstrates the AAV-mediated and MLC-2v regulated CD151 gene is highly expressed in the ischemic myocardium and cardiac-specific delivery that is more efficiently targets CD151 to the ischemia myocardium after myocardial infarction.
文摘目的对微光刀立体定向放射治疗系统(Dual Camera 2 mm Stereotactic Radiosurgerg,D2SRS)临床剂量学可靠性进行测试、研究。方法随机选取20例已接受放射治疗的病例,其中包括7例颅内肿瘤、6例肺部肿瘤和7例肝癌病例,将20例病例的CT图像和RT Structures通过DICOM网络分别传输至雷泰Ti GRT计划系统和Eclipse11.0计划系统,进行放射治疗计划设计,统计两组靶区及危及器官的剂量学参数并进行统计学分析,将两组调强计划分别进行绝对剂量和二维面剂量验证,并比较两组计划的绝对剂量和二维平面剂量验证结果。结果两组计划均能满足临床剂量要求,靶区剂量学参数D_(98)、D_(50)、D_(02)、CI以及HI均无统计学差异(P>0.05);危及器官受照剂量如全脑、心脏、食管、肝脏和肾脏的D_(mean)、脑干和脊髓D_(max)、双肺V_(20)等剂量学参数均无统计学差异(P>0.05);Eclipse与Ti GRT两组计划的验证结果显示,点剂量误差统计结果为(0.70±1.44,0.57±1.86,t=0.268,P=0.796);二维面剂量Gamma分析统计结果为(96.8%±0.9%,97.3%±1.2%,t=2.031,P=0.056)。结论D2SRS在剂量精确性上达到同类型进口设备相似水平。
