p53Mutations and Presence of HPV DNA Do Not Correlate with Radiosensitivity of Gynecological Cancer Cell Lines

Objective.The correlation betweenp53tumor suppressor gene mutations and the presence of high-risk human papillomavirus (HPV) DNA with thein vitroradiosensitivity of gynecological malignancies was studied in 26 cell lines derived from gynecological cancers of 23 patients.Methods.Comparison of the intrinsic radiosensitivity was performed with mean inactivation dose (D?) determined with the 96-well plate clonogenic assay.p53mutations were investigated with polymerase chain reaction and single-strand conformation polymorphism (PCR–SSCP) analysis and direct DNA sequencing, and the presence of HPV DNA was studied with PCR using HPV consensus primers.Results. p53mutations were found in 6 of 10 vulvar squamous cell carcinoma (SCC) lines. Nine vulvar and 1 vaginal SCC cell lines were HPV DNA negative and 1 vulvar cell line was HPV 16 positive. All 4 cervical SCC lines were HPV positive and possessed the wild-typep53.Three cell lines expressed HPV 16 and 1 HPV 68. Among 10 endometrial cancer cell lines, 2 cell lines with mutantp53and 1 HPV 16 positive cell line were found. No correlation could be demonstrated between inactivation of thep53gene and radiosensitivityin vitro;the cell lines were evaluated as one group or according to their anatomical origin or histology.Conclusion.Our results indicate that inactivation of thep53gene through mutation or binding with HPV DNA does not increase the resistance of gynecological malignancies to ionizing radiationin vitro.     

研究经络本质的新途径

目的:探索经络的物质本质的新途径.方法:回顾分析了国内外近20年经络实质研究概况,并详细介绍了缝隙连接(GJ)、半通道的结构特点及其与经络的某些相似之处.结果:针刺效应的产生与神经系统、内分泌系统、免疫系统以及结缔组织等密切相关;同一组织内和不同组织间相同和不同细胞间的GJ、半通道可能是机械性刺激信号(针刺)在体内转变为生物电信号或化学信号的结构基础.结论:针刺效应是在腧穴接受刺激的条件下,以高密度的GJ、半通道作为物质基础,以细胞间液、组织间液为缓冲系统的神经、血管、内分泌、免疫等多个系统的综合反应.     

胆酸钠对人卵巢癌细胞系作用的研究

目的 探讨胆酸钠单独应用及联合全反式维甲酸对2株卵巢癌细胞系(COC2、CAOV3)的体外抑制作用。方法 2003年5～8月对经不同浓度的胆酸钠及胆酸钠联合全反式维甲酸作用后的两株细胞采用MTT比色法测定细胞生长抑制率，流式细胞仪测定细胞周期变化，并行培养细胞的光镜、电镜观察。结果胆酸钠对两株细胞的抗增殖活性在50～150mg／L范围内呈现剂量依赖效应，并且在两药合用时此作用更明显；单用胆酸钠及两药联合应用均能随用药浓度而明显增加G0／G1期细胞比例，降低S、G2／M期比例，并以两药合用更为明显；光镜、电镜结果显示：胆酸钠100mg／L组与全反式维甲酸10μmol／L组细胞形态变化相似，但两者合用时细胞凋亡明显。结论 在两株卵巢癌细胞中胆酸钠不但具有单独而且有协同全反式维甲酸抑制肿瘤的作用，胆酸钠能明显降低全反式维甲酸的用量从而减少其副反应，并且不降低其抗肿瘤作用。     

细胞因子HGF体外增强骨肉瘤细胞株的增殖和黏附性

【目的】研究肝细胞生长因子，离散因子（HGF／SF）及其受体蛋白c-Met在骨肉瘤细胞株MG-63和HOS中的表达及对瘤细胞生物学行为的影响，探讨HGF／c-Met系统在骨肉瘤细胞生长侵袭过程中的作用。【方法】用流式细胞术检测HGF-α、HGF-β、c-Met在2株骨肉瘤细胞株中的表达；外源性HGF刺激此2株细胞株后，用MTT法、黏附性检测方法研究细胞增殖速率和黏附性的变化。【结果】HGF-α、HGF-β、c-Met在2株骨肉瘤细胞株中的阳性表达率均低于3％。HGF浓度达50ng／mL以上时．MG-63和HOS的增殖速率显著增高，P〈0．01。当外源性HGF的浓度分别达5ng／mL和10ng／mL时，MG-63和HOS的黏附性指标即显著升高，P〈0．01。外源性HGF刺激后HGF-α、HGF-β、c-Met在2株骨肉瘤细胞株中的表达略有变化，但差异无统计学意义，P〉0．05。【结论】HGF可以促进骨肉瘤细胞株MG-63和HOS的体外增殖和黏附能力。     

人肝癌细胞株VEGF/VEGFR的检测及意义探讨

目的：筛选血管内皮生长因子（vascular endothelial growth factor，VEGF）高表达肝癌细胞株，为进一步研究VEGF在肝癌治疗中的作用提供理论依据。并探讨VEGF受体在肝癌细胞株的表达及意义。方法：ELISA法及Western blot法分别检测人肝癌细胞株培养上清及细胞内VEGF蛋白的表达。免疫细胞化学方法检测VEGFR在人肝癌细胞中的表达。结果：5株肝癌细胞株均见VEGF蛋白表达，其中SMMC-7721的VEGF表达量最高，VEGF特异性受体Fit-1在HepG2、HHCC、SMMC-7721、Bcl-7402见阳性表达，KDR在HepG2、HHCC、Bel-7402、QGY-7701见阳性表达。结论：肝癌细胞株中VEGF表达量不尽一致，VEGF在肝癌发生发展中可能存在自分泌作用方式。     

熊果酸体外抗胃癌细胞SGC7901机制的研究

目的探讨熊果酸(urso lic ac id,UA)体外抑制胃癌细胞SGC 7901生长的作用机制。方法体外培养人胃癌细胞株SGC 7901,M TT法观察不同浓度UA作用不同时间对细胞生长的影响;不同浓度UA(0~40μm o l/L)处理SGC 7901细胞24 h后,倒置显微镜观察细胞形态变化;荧光染料Hoechst 33258染色和流式细胞仪检测细胞凋亡情况;W estern B lotting法检测凋亡相关蛋白B cl-2和B ax的表达。结果20~40μm o l/L UA可抑制SGC 7901的生长,并呈浓度和时间依赖性,其作用12、24、36、48 h的IC50分别为(57.50±1.18)、(34.28±2.05)、(27.54±1.11)、(24.83±1.02)μm o l/L。20~40μm o l/L UA作用24 h后,SGC 7901细胞变圆,出现不同程度的漂浮;同时细胞被阻滞于G0/G1期并发生凋亡,随药物浓度升高,凋亡率增加,凋亡相关蛋白B cl-2表达减少,B ax无明显变化。结论熊果酸对SGC 7901细胞具有较强的抗肿瘤活性,其机制可能与细胞毒作用、增殖抑制作用以及下调凋亡相关蛋白B cl-2表达而促进凋亡有关。     

姜黄素对人淋巴细胞低密度脂蛋白受体表达影响的研究

 目的建立一种简便的直接检测人类低密度脂蛋白受体(lowdensitylipoproteinrecepor,LDL-R)活性并筛选降脂中药的细胞模型:人B淋巴细胞永生化细胞系研究不同浓度姜黄素对人类LDL-R在人淋巴细胞中表达的影响,在细胞和受体水平上探讨姜黄素的作用机制。方法EBV转化技术建立的人B淋巴细胞永生化细胞系;以荧光试剂标记配体法,利用流式细胞仪技术和激光扫描共聚焦显微镜技术研究姜黄素对人淋巴细胞LDL-R表达的影响。结果姜黄素在5～50μmol·L^-1内可以增强人淋巴细胞LDL-R的表达,并且具有明显的量效关系。结论利用荧光试剂标记配体法检测人淋巴细胞LDL-R活性是一个简单有效的方法;姜黄素是一个非常强的LDL-R基因表达促进剂,可能通过增加人淋巴细胞LDL-R的表达起到降血脂和抗动脉粥样硬化的作用。     

环氧化酶-2抑制剂nimesulide对乳腺癌细胞株增生、凋亡的影响

目的 探讨环氧化酶-2抑制剂nimesulide(NIM)对雌激素受体(ER)阴性(MDA-MB．231)和阳性(MCF-7)人乳腺癌细胞株增生、凋亡的影响。方法 应用MTT比色法分析细胞生长抑制作用，流式细胞技术测定细胞周期分布和凋亡率，透射电镜观察细胞形态与超微结构，AnnexinV法检测细胞的凋亡。结果 NIM以时间、剂量依赖性方式抑制MDA-MB-231(COX-2阳性)、MCF-7(COX-2阴性)细胞生长，阻滞细胞周期于G0／G1期，诱导细胞的凋亡，MDA—MB-231细胞对NIM的作用更为敏感。NIM对COX-2表达阴性的MCF-7细胞同样具有抑制增生、诱导凋亡的作用。结论 NIM对ER阳性和ER阴性乳腺癌细胞均有抑制增生、诱导凋亡的作用。NIM的抗肿瘤作用存在环氧化酶．2依赖性与非依赖性两种途径。     

Achieving Near-Zero and Zero: Who Said Interventions and Controls Don't Matter?

BackgroundImplementation of the Institute for Healthcare Improvement's Central Line Bundle in 2005 did not result in attainment of 0 central line-associated bloodstream infections (CLABSIs).PurposeTo establish process improvements to eliminate CLABSIs.MethodsInfection Prevention and Intravenous Therapy Departments at our institution formed a collaborative partnership in 2008 to eliminate CLABSIs. Staff education, daily surveillance, and implementing evidence-based practice into policies, protocols, and product selection were utilized. A study designed to capture and track multiple interventions from the beginning versus time was implemented.ResultsThere were a total of 7 CLABSIs over 5 years, which trended down by year from 3 to 2 to 1 to 1 to 0. The average annual CLABSI rate of infections per 1,000 catheter line days declined from 1.068 to 0.849 to 0.508 to 0.449 to 0.000. The full 3-year simple average from 2009 to 2011, after all of the key interventions were phased-in, was 0.3048.ConclusionsCollaboration and commitment played a role in implementing changes. Empowering staff through education that increases knowledge, understanding, and skills improves patient safety and patient outcomes. Process changes and practice protocols supported by evidence-based practice along with efficacious products played a crucial part in getting to 0 CLABSIs. Layered kits with products placed in the correct order of use can increase aseptic technique compliance. Prompt identification of individual CLABSI episodes with prompt intervention to the specific individuals involved has been shown to be cost-effective and can be implemented in most hospitals.