背景与目的:考察纳米级阳离子聚酰胺-胺型树枝状聚合物(polyam idoam ine dendrimers,PAMAM-D)作为基因载体进行前列腺癌自杀基因治疗的可行性,为前列腺癌的基因治疗寻找新的基因载体。方法:以第5代PAMAM-D(G5-PAMAM-D)为载体将含增强...背景与目的:考察纳米级阳离子聚酰胺-胺型树枝状聚合物(polyam idoam ine dendrimers,PAMAM-D)作为基因载体进行前列腺癌自杀基因治疗的可行性,为前列腺癌的基因治疗寻找新的基因载体。方法:以第5代PAMAM-D(G5-PAMAM-D)为载体将含增强型荧光蛋白(EGFP)基因片段的重组质粒pEGFP-C1转染至前列腺癌细胞系PC-3和22Rv1,成功表达EGFP后,再以G5-PAMAM-D为载体将含HSV-tk自杀基因的真核表达重组质粒pcDNA3-tk转染至前列腺癌细胞系PC-3和22Rv1,转染48h后,对转染的两种细胞给予浓度为0、10、100、1000、10000μmol/L前体药更昔洛韦(Ganciclovir,GCV),24h后采用MTT比色法测定药物对细胞增殖的影响。制作前列腺癌皮下荷瘤小鼠,将G5-PAMAM-D/pcDNA3-tk复合物瘤内注射,24h后腹腔注射GCV,观察这一复合物体内对肿瘤生长的抑制作用。结果:荧光观察及FCM结果证明G5-PAMAM-D可将pEGFP-C1转入两种前列腺癌细胞并表达EGFP。采用G5-PAMAM-D将pcDNA3-tk重组质粒转染PC-3和22Rv1细胞,给不同浓度的前体药GCV后,实验组细胞与对照组相比有明显浓度依赖性生长抑制。荷瘤小鼠在给予瘤内注射G5-PAMAM-D/pcDNA3-tk复合物后,行腹腔注射GCV,治疗后第40天,实验组肿瘤体积(1135±245mm3)与对照组裸质粒组(9965±2109mm3)和PAMAM-D组(8357±1956mm3)相比肿瘤生长明显被抑制(P<0.01),生存时间延长。结论:G5-PAMAM-D可作为前列腺腺癌自杀基因治疗的基因载体,有良好的应用前景。展开更多
Endotoxin is lipopolysaccharide(LPS) derived from the cell membrane of gram-negative bacteria. The concentration of endotoxin in endotoxemia patients′ blood increases quickly and leads to severe sepsis, leading to se...Endotoxin is lipopolysaccharide(LPS) derived from the cell membrane of gram-negative bacteria. The concentration of endotoxin in endotoxemia patients′ blood increases quickly and leads to severe sepsis, leading to severe hypertension, cardiovascular continues to have a high failure and death, which is a major cause of death in patients and continues to have a high mortality despite appropriate surgery, potent antibiotic and intensive supportive therapy was conducted. Removal of endotoxin duly and effectually from patients′ blood is very important in clinic. Hemoperfusion can clear toxins from patient′s blood by adsorbent, not only small molecular toxins but also large ones in time. In this paper, the hemoperfusion adsorbent for removing endotoxin was prepared. The physical and chemical parameter of the adsorbent is investigated and the surface structure of adsorbent was investigated through ESEM. The influence of weights of adsorbents and adsorbing time on adsorption ability was also studied. The results showed that the adsorbent with lysine as the ligand had good affinity to endotoxin and the best adsorption capacity of it was 253.1 EU/g when the initial concentration is 10 EU/mL. The blood compatibility of the adsorbent was also studied and the results indicated that the adsorbent has clinic application foreground.展开更多
文摘背景与目的:考察纳米级阳离子聚酰胺-胺型树枝状聚合物(polyam idoam ine dendrimers,PAMAM-D)作为基因载体进行前列腺癌自杀基因治疗的可行性,为前列腺癌的基因治疗寻找新的基因载体。方法:以第5代PAMAM-D(G5-PAMAM-D)为载体将含增强型荧光蛋白(EGFP)基因片段的重组质粒pEGFP-C1转染至前列腺癌细胞系PC-3和22Rv1,成功表达EGFP后,再以G5-PAMAM-D为载体将含HSV-tk自杀基因的真核表达重组质粒pcDNA3-tk转染至前列腺癌细胞系PC-3和22Rv1,转染48h后,对转染的两种细胞给予浓度为0、10、100、1000、10000μmol/L前体药更昔洛韦(Ganciclovir,GCV),24h后采用MTT比色法测定药物对细胞增殖的影响。制作前列腺癌皮下荷瘤小鼠,将G5-PAMAM-D/pcDNA3-tk复合物瘤内注射,24h后腹腔注射GCV,观察这一复合物体内对肿瘤生长的抑制作用。结果:荧光观察及FCM结果证明G5-PAMAM-D可将pEGFP-C1转入两种前列腺癌细胞并表达EGFP。采用G5-PAMAM-D将pcDNA3-tk重组质粒转染PC-3和22Rv1细胞,给不同浓度的前体药GCV后,实验组细胞与对照组相比有明显浓度依赖性生长抑制。荷瘤小鼠在给予瘤内注射G5-PAMAM-D/pcDNA3-tk复合物后,行腹腔注射GCV,治疗后第40天,实验组肿瘤体积(1135±245mm3)与对照组裸质粒组(9965±2109mm3)和PAMAM-D组(8357±1956mm3)相比肿瘤生长明显被抑制(P<0.01),生存时间延长。结论:G5-PAMAM-D可作为前列腺腺癌自杀基因治疗的基因载体,有良好的应用前景。
文摘Endotoxin is lipopolysaccharide(LPS) derived from the cell membrane of gram-negative bacteria. The concentration of endotoxin in endotoxemia patients′ blood increases quickly and leads to severe sepsis, leading to severe hypertension, cardiovascular continues to have a high failure and death, which is a major cause of death in patients and continues to have a high mortality despite appropriate surgery, potent antibiotic and intensive supportive therapy was conducted. Removal of endotoxin duly and effectually from patients′ blood is very important in clinic. Hemoperfusion can clear toxins from patient′s blood by adsorbent, not only small molecular toxins but also large ones in time. In this paper, the hemoperfusion adsorbent for removing endotoxin was prepared. The physical and chemical parameter of the adsorbent is investigated and the surface structure of adsorbent was investigated through ESEM. The influence of weights of adsorbents and adsorbing time on adsorption ability was also studied. The results showed that the adsorbent with lysine as the ligand had good affinity to endotoxin and the best adsorption capacity of it was 253.1 EU/g when the initial concentration is 10 EU/mL. The blood compatibility of the adsorbent was also studied and the results indicated that the adsorbent has clinic application foreground.