结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

目的研究高效结肠癌富伴侣分子疫苗的制备及其抗癌治疗作用。方法结肠癌CT26细胞在0.1mg／L天花粉蛋白的培养基中不同温度下培养.诱导癌细胞伴侣分子的表达和抗原肽的合成。超声裂解并离心,盐析法分离上清蛋白,低温透析去除相对分子质量低于50000和高于300000的蛋白分子。凝胶过滤结合变性聚丙烯酰胺凝胶电泳法（SDS.PAGE）收取相对分子质量70000、90000、95000、110000和170000处峰段蛋白。对所获蛋白定性定量,检测不同蛋白复合物的T细胞增殖、抗结肠癌自然杀伤细胞（NK）和细胞毒性T淋巴细胞（CTL）活性等主要抗癌效应。结果主要伴侣分子（HSP60、HSP70、HSP90、gp96、HSP110和HSP170）的抗原肽复合物被较好提取.制备了浓缩的富伴侣分子抗原肽。体外抗癌实验显示,相对于普通处理的瘤苗,各肠癌瘤苗的细胞增殖、NK和CTL细胞活性等主要抗癌效应均有明显提高（P〈0.01）。结论凝胶过滤结合SDS-PAGE法可以更好地提取制备浓缩的富伴侣分子抗原肽;热应激和天花粉蛋白处理可使抗原肽的表达量增加和免疫原性提高。从而诱导出更强的抗癌免疫,其中42℃热应激加天花粉蛋白组瘤苗的抗癌作用最强。     

2型糖尿病患者CD11b/CD18的表达及活血化瘀药对其的影响

目的：探讨在2型糖尿病(DM)患者中多形核白细胞粘附分子CDllb／CD18的表达及活血化瘀药预防血管病变的可能机制。方法：49例无高血压、无临床肾病的2型DM患者随机分成中药治疗组(26例)和常规治疗组(23例),治疗3个月。治疗前后观察尿白蛋白排泄率、CDllb／CD18的表达及肿瘤坏死因子-α浓度等指标。结果：与健康对照组比较,2型DM患者的CDllb／CD18表达增高,肿瘤坏死因子-α浓度也明显增高(P＜0．01)。与常规治疗组比较,活血化瘀中药治疗3个月后,2型DM患者的CDllb／CD18表达,尿白蛋白排泄率和肿瘤坏死因子-α明显降低(P＜0．01),而常规治疗组无类似的变化。相关性分析提示,尿白蛋白排泄率下降与肿瘤坏死因子-α浓度降低和多形核白细胞粘附分子CDllb／CDl8表达的下降呈正相关(r＝0．56,P＜0．01,r＝0．64,P＜0．01)。结论：2型DM患者的多形核白细胞粘附分子CDllb／CD18表达增高;活血化瘀法预防血管病变的机制可能与其抑制CDllb／CD18表达有关。     

2型糖尿病患者CD11b/CD18的表达及活血化瘀药对其的影响

目的:探讨在2型糖尿病(DM)患者中多形核白细胞粘附分子CD11b/CD18的表达及活血化瘀药预防血管病变的可能机制.方法:49例无高血压、无临床肾病的2型DM患者随机分成中药治疗组(26例)和常规治疗组(23例),治疗3个月.治疗前后观察尿白蛋白排泄率、CD11b/CD18的表达及肿瘤坏死因子-α浓度等指标.结果:与健康对照组比较,2型DM患者的CD11b/CD18表达增高,肿瘤坏死因子-α浓度也明显增高(P＜0.01).与常规治疗组比较,活血化瘀中药治疗3个月后,2型DM患者的CD11b/CD18表达,尿白蛋白排泄率和肿瘤坏死因子-α明显降低(P＜0.01),而常规治疗组无类似的变化.相关性分析提示,尿白蛋白排泄率下降与肿瘤坏死因子-α浓度降低和多形核白细胞粘附分子CD11b/CD18表达的下降呈正相关(r＝0.56,P＜0.01,r＝0.64,P＜0.01).结论:2型DM患者的多形核白细胞粘附分子CD11b/CD18表达增高;活血化瘀法预防血管病变的机制可能与其抑制CD11b/CD18表达有关.     

72例急性白血病免疫学与细胞遗传学研究

目的评价免疫学与细胞遗传学联合检测在急性白血病（AL）分型中的意义。方法采用CD45/SSC双参数散点图设门方法进行三色/四色流式细胞术免疫表型分析、R显带技术进行染色体分析。结果形态学分型与免疫学检查符合率为89.0%,有2例形态学拟诊为M7亚型的AML经免疫分型确诊,3例形态学拟诊为M3亚型的AML经染色体检查除外M3亚型;AML患者较特异性抗原表达为CD13（90.0%）,CD33（94.0%）,染色体异常率55%,其中特异性染色体异常占59%。伴t（15;17）染色体易位M3特异性表达CD13（100%）、CD33（100%）。M2b特异性表达CD19（100%）、CD34（100%）与t（8;21）染色体易位（100%）。M5易有CD14（25%）表达与t/del（11）（q23）染色体异常（50%）。髓系中易见淋系相关抗原表达,常见为CD7（24.0%）,CD19（12%）,CD10（6.0%）。CD7阳性AML患者染色体异常率达73%,显著高于CD7阴性AML患者（P〈0.05）;B-ALL患者染色体异常率83%,特异性染色体异常率53%。T-ALL患者染色体异常率71%,特异性染色体异常率60%。淋系中亦可合并髓系抗原表达,常见为CD13（52.0%）,CD33（35%）。结论免疫学分型结合染色体检查可提高AL的诊断准确性、判断预后、揭示细胞生物学特征。特殊类型AL如M0、M3、M7等的诊断必须依赖免疫分型与染色体检查。     

脐血造血干细胞分离方法的比较

目的：探讨分离脐血造血干细胞的最佳方法。方法：采用Ｆｉｃｏｌ分层法、３％明胶自然沉降法及６％羟乙基淀粉自然沉降法。结果：３％明胶自然沉降法所获有核细胞（ＮＣ）密度高达９４．６０（±２３．８０）×１０５／ｍｌ,ＣＦＵＧＭ数为６４．３０（±１２．１８）／１０５ＮＣ,ＣＤ３４＋细胞数为１．２１（±０．３７）×１０５／ｍｌ,均较其它两种方法为高（Ｐ＜０．０５）。结论：３％明胶自然沉降法是一种较理想的分离脐血造血干细胞的方法。     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

CD4+CD25+调节性T细胞与HIV感染关系的研究进展

CD4 CD25 调节性T细胞是一类具有特殊免疫调节功能的T细胞亚群。它能够抑制自身免疫病的发生,参与肿瘤免疫的调节,在感染和移植免疫中也发挥着重要作用。T细胞的这一亚群具有免疫无能和免疫抑制特性,新近发现它亦与艾滋病的发生、发展关系密切。文中就CD4 CD25 调节性T细胞与HIV感染的研究进展进行综述。     

大萼香茶菜甲素通过线粒体途径诱导HL-60白血病细胞凋亡

目的：观察大萼香茶菜甲素（macrocalyxinA,MA）体外诱导HL-60细胞凋亡,并探讨其作用机制。方法：不同浓度的MA与HL-60细胞进行培养,采用MTT比色法观察其对HL-60细胞增殖的抑制作用;细胞形态学、DNA含量、DNA梯度电泳及细胞周期分析、Annexin—V／PI双标记和Hoechst 33258荧光染色等分析其促细胞凋亡效应;流式细胞术和RT-PCR分别检测Bcl-2、Bax、P53、Fas、线粒体膜蛋白（Ap02．7）、线粒体跨膜电位（AWm）与Bcl-2、Bax、P53、caspase-3 mRNA变化水平,研究其促凋亡机制。结果：MA呈现作用时间和剂量依赖性地抑制HL-60细胞增殖和活力;HL-60细胞经MA作用后,Wright—Giemsa染色和Hoechst荧光染色后细胞出现典型的凋亡小体,细胞阻滞于G0／G1期,DNA片段化,亚二倍体明显增高,Annexin—V／PI标记升高;MA诱导HL-60细胞凋亡过程中,Bcl-2、Fas、P53表达无明显变化,Bax、线粒体膜蛋白（Apo2．7）、caspase-3表达显著增加,Bax／Bcl-2比值升高,龇下降。结论：MA能抑制HL-60细胞增殖和细胞活力、诱导细胞凋亡,其机制通过上调Bax基因和Bax／Bcl-2比值,使线粒体膜电位下降、膜通透性增高,最终使caspase-3激活而促进凋亡。     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.