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The growth inhibiting rate of T24 cell lines were detected by MTT methods, apoptosis of cells were detected by flow cytometry, the mechanism of apoptosis was analyzed by detecting the protein expression of Bcl-2, Bax, Caspase-9, Caspase-3 and cytoplastic protein Cytochrome C. 4 We injected live T24 cells into the subcutaneous space of nude mice and successfully built up the animal model of bladder carcinoma. The effect of CS-PAA-EPI polymer magnetic microspheres targeting chemotherapy was investigated by HE staining, TUNEL ,tumor weight and volume inhibition rate. Results: 1 TEM revealed that the CS-PAA polymer magnetic microspheres were regular spherical shape,the average diameter was 80nm in dry condition. By controlling the pH value of the medium,polymers had positive or negative zeta potential. VSM showed the CS-PAA polymer magnetic microspheres had superparamagnetic. The diameter of CS-PAA-EPI polymer magnetic microspheres were 200nm in solution by DLS examining,the embedding ratio was 20%,the EPI loading rate was 15%, which was higer than reported in other articles. 2 Raw eye observation found that the rat"s bladder of treatment group was brown color,which meaned the aggregation of iron particles, compared with the control group, iron stain found iron particles were assembled in rat"s bladder of the treatment group, the amount of iron particles in liver and spleen were less obviously.

研究结果:1合成的CS-PAA磁性聚合物微球呈球形,大小均一,TEM测定其干态下粒径为80nm左右,磁化曲线证实具有超顺磁性,具有一定的PH敏感性,固载表柔比星后,水溶液性状稳定,无沉淀物,DLS测定直径约200nm左右,测定载药率为15%,较文献报道高,包封率为20%。2肉眼观察试验组大鼠膀胱表面呈褐色,可见大量的Fe粒子聚集,普鲁士兰染色法显示,试验组大鼠膀胱壁内有大量的Fe粒子,分布至膀胱壁全层,与对照组大鼠相比,试验组大鼠的肝、脾内的Fe粒子聚集量明显降低;HPLC测定结果与Fe染色相同;高剂量磁性CS-PAA-EPI生理盐水组及单纯EPI生理盐水组均在给药后14天出现血肌酐和尿素氮的升高,其他组大鼠血生化指标没有明显变化。3MTT法发现,高、中、低剂量磁性CS-PAA-EPI生理盐水组在外加磁场的协同作用下杀伤T24细胞效应明显高于单纯的EPI生理盐水组,FCM发现试验药物组可引起明显的肿瘤细胞凋亡,试验药物治疗组细胞胞浆内出现了由线粒体释放出的细胞色素C,试验组细胞Bcl-2蛋白减少,Bax蛋白变化不明显,Caspase-3、Caspase-9蛋白受到了激活活化。4高、中、低剂量磁性CS-PAA-EPI生理盐水组的瘤重抑制率和瘤体积抑制率均明显高于单纯的EPI生理盐水组(P<0.01),其中高剂量组的抑制率最高。

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