hSav1蛋白高表达对Mst1介导的细胞凋亡的影响

Objective To elucidate the effect of hSavl expression on Mstl-mediated apoptosis in HeLa cells. Methods Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and cotransfected into HeLa cells. Triple immunofluorescent labeling of hSav1, Mst1 and nucleus was performed to determine their subcellular localization. Plasmids pCMV-HA-hSav1 and/or pcDNA/4TO-Flag-Mst1 were transfected into HeLa cells, and 36 hours later cisplatin (50 μmol/L) as a pro-apoptotic agent was added for 14 hours. Cell apoptosis was analyzed by annexin V/PI assay. Results Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and the authenticity of constructs was verified by sequencing. The binding in vitro showed that hSavl could be detect from the anti-Mstl immunoprecipitation complex. The immunofluorescent labeling showed that hSavl and Mstl had the same localization in cells. Overexpressed protein hSavl did not induce a significant cell apoptosis. However, co-expression of hSavl with Mstl resulted in a significant increase of apoptosis above the level seen with Mstl alone (24. 5% ± 2.4% vs. 39.3% ± 4.0%, P < 0. 05). Conclusion Our findings indicate that hSavl is a newly identified protein that interacts with Mst1 and a, augments Mst1-mediated apoptosis.     

hSav1蛋白高表达对Mst1介导的细胞凋亡的影响

Objective To elucidate the effect of hSavl expression on Mstl-mediated apoptosis in HeLa cells. Methods Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and cotransfected into HeLa cells. Triple immunofluorescent labeling of hSav1, Mst1 and nucleus was performed to determine their subcellular localization. Plasmids pCMV-HA-hSav1 and/or pcDNA/4TO-Flag-Mst1 were transfected into HeLa cells, and 36 hours later cisplatin (50 μmol/L) as a pro-apoptotic agent was added for 14 hours. Cell apoptosis was analyzed by annexin V/PI assay. Results Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and the authenticity of constructs was verified by sequencing. The binding in vitro showed that hSavl could be detect from the anti-Mstl immunoprecipitation complex. The immunofluorescent labeling showed that hSavl and Mstl had the same localization in cells. Overexpressed protein hSavl did not induce a significant cell apoptosis. However, co-expression of hSavl with Mstl resulted in a significant increase of apoptosis above the level seen with Mstl alone (24. 5% ± 2.4% vs. 39.3% ± 4.0%, P < 0. 05). Conclusion Our findings indicate that hSavl is a newly identified protein that interacts with Mst1 and a, augments Mst1-mediated apoptosis.     

B超定位前列腺扩张术的临床应用

Ｂ超定位前列腺扩张术的临床应用宝应县人民医院泌尿外科沈开福，徐建林宝应县人民医院Ｂ超室韩春芳，李兆明我院自１９９０年开始应用前列腺扩张术有选择地对前列腺增生症患者进行治疗。５年来，我们从用手指在肛门内定位以及Ｘ光下定位过渡到用Ｂ超导航定位，其定位准确...     

Inhibitory Effects of Mild Hyperthermia plus Docetaxel Therapy on ER（＋/-） Breast Cancer Cells and Action Mechanisms

Summary： The purpose of this study was to verify that a combination of mild hyperthermia and do- cetaxel chemotherapy produces synergistic antitumor effects and to explore the action mechanisms of this treatment approach. The effects of docetaxel on the proliferation of cells from the estrogen receptor （ER）-positive human breast cancer cell line MCF-7 and the ER-negative human breast cancer cell line MDA-MB-453 were examined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay, and effective experimental concentrations of docetaxel were determined. The effects of mild hy- perthermia plus docetaxel therapy on apoptosis rate in the MCF-7 and MDA-MB-453 human breast cancer cell lines were analyzed by using flow cytometry with Annexin-V fluorescein isothiocyanate （FITC）/propidium iodide （PI） staining. The effects of these combined treatments on cell cycle progres- sion in the MCF-7 and MDA-MB-453 human breast cancer cell lines were examined by using flow cy- tometry. The effects of these combined treatments on the expression of apoptosis-related proteins and proteins in the mitogen-activated protein kinase （MAPK） pathways were analyzed by using Western blotting. The effects of these combined treatments on the expression of the heat shock protein 70 （HSP70） and the multi-drug resistance （MDR） gene product P-glycoprotein （Pgp） were examined by using Western blotting. The results showed that the half-maximal inhibitory concentration （IC50） of do- cetaxel for MCF-7 and MDA-MB-453 cells was 19.57±1.12 and 21.64±2.31 gmol/L respectively. Mild hyperthermia with docetaxel therapy could increase apoptosis rate in the MCF-7 and MDA-MB-453 cells. Apoptosis rate in MCF-7 and MDA-MB-453 cells was increased from （23.66±3.59）% and （18.51±3.17）% in docetaxel treatment group to （47.12±6.73）% and （55.16±7.42）% in mild hyperthermia plus docetaxel group, indicating that the mild hyperthermia and docetaxel therapeutic approaches exhib- ited significant synergistic antitumor effects. Treatments of mild hyperthermia plus docetaxel induced G2/M cell cycle arrest in the MCF-7 and MDA-MB-453 cells. Western blotting demonstrated that pro- teins in the MAPK pathway were expressed at higher levels in docetaxel-treated cells following mild hypothermia than those in cells treated with docetaxel alone. As compared with blank control group, cells from the mild hyperthermia plus docetaxel group exhibited significantly decreased B-cell lym- phoma 2 （Bcl-2） protein expression but slightly increased Bcl-2-associated X protein （Bax） expression. Western blotting results revealed that HSP70 and Pgp expression levels were significantly increased following mild hypothermia. It was concluded that treatments of mild hyperthermia plus docetaxel in- hibited the proliferation of human breast cancer cells, promoted apoptosis of breast cancer cells, and produced synergistic antitumor effects.     

姜黄素调控乳腺癌细胞增殖与转移机制研究

目的研究姜黄素对人乳腺癌细胞MDA MB 453中缺氧诱导因子 1α（HIF 1α）的影响及其对肿瘤细胞增殖和转移的作用。方法采用噻唑蓝（MTT）法和细胞计数观察姜黄素抑制MDA MB 453细胞增生作用;采用transwell侵袭小室测定姜黄素对MDA MB 453转移的影响;采用Real time PCR和Western blot印迹法检测常氧和乏氧状态下姜黄素对MDA MB 453细胞系中HIF 1α的mRNA和蛋白表达水平的影响,同时采用Real time PCR和ELISA法检测姜黄素对HIF 1α下游靶基因血管内皮生长因子（VEGF）mRNA水平和蛋白表达水平的影响。结果姜黄素对MDA MB 453细胞体外生长和转移具有抑制作用;常氧、乏氧状态下加入姜黄素后,MDA MB 453细胞系中HIF 1α的mRNA水平无变化,但蛋白水平明显降低,VEGF mRNA水平和蛋白水平均明显降低。结论姜黄素通过HIF 1α调控MDA MB 453细胞系增殖和转移。     

高表达GRIM-19诱导结肠癌SW480细胞凋亡

目的：研究干扰素/维甲酸诱导死亡基因（retinoidinterferoninduced mortality,GRIM19）对结肠癌SW480细胞凋亡的影响。方法：构建GRIM19真核表达载体（pCMVFlagGRIM19）,转染入SW480细胞中,Western blotting检测GRIM19及凋亡相关蛋白Balxl的表达,采用AnnexinV/PI和线粒体膜电位JC-1染色检测SW480细胞的凋亡。结果：成功构建pCMVFlagGRIM19真核表达载体。pCMV-Flag-GRIM-19质粒转染SW480细胞后,GRIM19的表达上调,凋亡抑制蛋白Bclxl的表达则下调。转染空质粒pCMVFlag组SW480细胞凋亡率为（7.7±1.39）%,转染pCMV-Flag-GRIM-19质粒后SW480细胞凋亡率为（15.0±2.52）%（P<0.05）。线粒体膜电位检测显示转染空质粒pCMVFlag组SW480细胞膜电位降低细胞为（7.5±2.09）%,而转染pCMVFlagGRIM19后细胞线粒体膜电位降低细胞为（17.5±3.07）%（P<0.05）。结论：GRIM-19体外转染可有效诱导结肠癌SW480细胞凋亡。     

hSav1蛋白高表达对Mst1介导的细胞凋亡的影响

Objective To elucidate the effect of hSavl expression on Mstl-mediated apoptosis in HeLa cells. Methods Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and cotransfected into HeLa cells. Triple immunofluorescent labeling of hSav1, Mst1 and nucleus was performed to determine their subcellular localization. Plasmids pCMV-HA-hSav1 and/or pcDNA/4TO-Flag-Mst1 were transfected into HeLa cells, and 36 hours later cisplatin (50 μmol/L) as a pro-apoptotic agent was added for 14 hours. Cell apoptosis was analyzed by annexin V/PI assay. Results Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and the authenticity of constructs was verified by sequencing. The binding in vitro showed that hSavl could be detect from the anti-Mstl immunoprecipitation complex. The immunofluorescent labeling showed that hSavl and Mstl had the same localization in cells. Overexpressed protein hSavl did not induce a significant cell apoptosis. However, co-expression of hSavl with Mstl resulted in a significant increase of apoptosis above the level seen with Mstl alone (24. 5% ± 2.4% vs. 39.3% ± 4.0%, P < 0. 05). Conclusion Our findings indicate that hSavl is a newly identified protein that interacts with Mst1 and a, augments Mst1-mediated apoptosis.     

结直肠癌中T-cadherin分子表达异常及意义

目的 探讨在结直肠癌组织中T-cadherin分子的表达及其与临床病理特征的关系.方法 应用免疫组化技术(SABC)对手术切除经病理证实的50例结直肠癌组织与癌旁组织中的T-cadherin分子表达进行分析.结果 50例结直肠癌组织中的T-cadherin分子表达阳性率为20%(10/50),50例癌旁组织中T-cadherin分子的表达阳性率为62%(31/50),两者差异有显著性意义(P<0.05).T-cadherin分子在结直肠癌中的表达情况与肿瘤的Dukes分期及淋巴结转移有密切的关系.结论 在结直肠癌组织中存在T-cadherin分子表达的缺失或低水平表达,而且随着肿瘤的恶性进展,T-cadherin分子的表达进行性下降.提示T-cadherin分子表达异常在结直肠癌发生发展过程中有重要作用.     

超声显像对脾组织块自体移植术后的随访

目的：探讨超声显像对脾组织块自体移植术后的随防的价值。方法：本组脾切除术后脾组织块自体移植术后10例，超声均测及脾块，经免疫学检查结果证实存活。结果：脾块均位于腹腔浅表部位，声像图特征：（1）脾组织块回声与正常脾相似，呈均匀低回声；（2）脾块包膜回声增厚；（3）长时间观察无变化。免疫学中，进行IgG、A、M、及C30测定，发现上述指标逐步升高，直到恢复正常，血常规正常，血中未发现Howell-Jolly小体及空泡红细胞。结论：超声显像可替代其他影像诊断方法，显示脾组织块。     

彩超在胎儿胆石病中的应用并文献复习

胎儿胆石病是非常罕见的 ,据英文文献记载目前仅有 30余例 [1 ] ,其中部分病例失随访或资料不完整。本文就我院自 1998年至今 ,5例资料完整的病例 ,报告如下 ,并结合文献资料的复习 ,以期提高对本病的认识和进一步研究。资料与方法我科自 1998年以来 ,经彩超检查晚期孕妇 30 0 0例 ,年龄 2 3～ 2 9岁 ,平均 2 4 .8岁 ,孕龄 32～ 4 0周 ,平均 32 .2周 ,均为孕 1产 0 ,共检出胎儿胆石病 5例 ,均为正常体检发现 ,无特殊药物应用史。在确定胎儿胆石病后 ,进行一系列实验室检查 ,包括母体和婴儿肝功能 ,及产后彩超复查。所用仪器为 HP尖端影像彩…