反义c-myc寡核苷酸对骨肉瘤细胞MG-63增殖的影响及机制

目的探讨反义c-myc寡核苷酸对人骨肉瘤MG-63细胞增殖的影响及机制。方法采用反义核酸技术设计反义c-myc寡核苷酸片段,转染入人骨肉瘤MG-63细胞,MTT法及流式细胞仪法检测瘤细胞体外增殖、细胞周期及c-myc基因蛋白表达情况。结果MTT法示人骨肉瘤MG-63细胞的增殖受抑,10．0μmol／L、作用48h效果最明显;流式细胞仪示反义c—myc寡核苷酸主要阻止人骨肉瘤MG-63细胞进入S期,c-myc基因蛋白表达受抑。结论反义c-myc寡核苷酸可抑制人骨肉瘤MG-63细胞的增殖;其机制为在基因水平干扰c-myc基因蛋白的表达。推测用反义策略抑制癌基因表达,或用基因敲除、基因替换的方法去除癌基因,可达到基因治疗的目的。     

同型半胱氨酸促血管平滑肌细胞增殖的机制

目的　研究同型半胱氨酸 (HCY)对血管平滑肌细胞 (VSMC)细胞周期、细胞周期素和 P2 7蛋白表达的影响。方法　用流式细胞技术测定细胞周期、细胞周期素和 P2 7蛋白表达量。结果　 HCY促进 VSMC增殖和细胞 S期合成 ,同时使细胞周期素 D、E、A表达明显增加 ,P2 7蛋白表达下降。结论　 HCY促进 VSMC增殖和细胞 S期合成可能是通过增加细胞周期素 D、E、A的表达 ,抑制 VSMC中 P2 7蛋白的表达来实现的。     

姜黄素对血管平滑肌细胞增殖及cyclinD1和p21wafl/cipl蛋白表达的影响

目的 观察姜黄素(curcumin)抑制大鼠血管平滑肌细胞(VSMC)增殖和诱导细胞周期停滞的作用,以及对细胞周期蛋白cyclinDl,p21wafl/cipl表达的影响.方法 采用组织贴块法体外培养大鼠VSMC,MTT法测定姜黄素对VSMC增殖的抑制作用,流式细胞仪分析细胞周期分布,Western印迹法检测cyclinDl,p21wafl/cipl蛋白的变化.结果 MTT示不同浓度姜黄素(7.5～120 μmol/L)在24～72 h范围内,浓度和时间依赖性抑制VSMC增殖;30 μmol/L以上浓度姜黄素显著抑制增殖的VSMC细胞周期进程,使S及G2/M期减少(P<0.05),G0/G1期增加(P<0.05);30 μmol/L的姜黄素可抑制cyclinDl表达,促进p21wafl/cipl蛋白表达.结论 姜黄素具有明确的抑制VSMC增殖和细胞周期停滞的作用,其与cyclinDl,p21wafl/cipl蛋白变化有关.     

周期素激酶抑制剂p27在肿瘤坏死因子-α诱导系膜细胞增生中的作用

目的 研究周期素激酶抑制剂p27在肿瘤坏死因子－α（TNF－α）诱导系膜细胞（MC）增生中的作用。方法 采用蛋白印迹（Western杂交）方法测定MC裂解液p27蛋白水平。[^3H]胸腺嘧啶核苷（[^3H]TdR）测定MC增生的情况,观察p27反义寡核苷酸（ODN）转染对TNF-α刺激MC中的p27水平及其对增生程度的影响。结果 TNF－α（200000U/L）可使无血清培养24h的MC中的p27水平降低（P＜0.01）,同时使[^3H]TdR掺入增加（P＜0.01）;p27反义ODN转染可降低TNF－α刺激24h的MC中的p27水平（P＜0.01）,同时可使[^3H]TdR掺入增加更为明显（P＜0.05）。结论 p27水平降低可能在TNF－α诱导MC增生中起重要作用。     

姜黄素对血管平滑肌细胞增殖及cyclinD1和p21waf1/cip1蛋白表达的影响

目的 观察姜黄素（curcumin）抑制大鼠血管平滑肌细胞(VSMC)增殖和诱导细胞周期停滞的作用,以及对细胞周期蛋白cyclinD1,p21waf1/cip1表达的影响。方法 采用组织贴块法体外培养大鼠VSMC,MTT法测定姜黄素对VSMC增殖的抑制作用,流式细胞仪分析细胞周期分布,Western印迹法检测cyclinD1,p21waf1/cip1蛋白的变化。结果 MTT示不同浓度姜黄素(7.5～120 μmol/L)在24～72 h范围内,浓度和时间依赖性抑制VSMC增殖;30 μmol/L以上浓度姜黄素显著抑制增殖的VSMC细胞周期进程,使S及G2/M期减少(P<0.05),G0/G1期增加(P<0.05);30 μmol/L的姜黄素可抑制cyclinD1表达,促进p21waf1/cip1蛋白表达。结论 姜黄素具有明确的抑制VSMC增殖和细胞周期停滞的作用,其与cyclinD1,p21waf1/cip1蛋白变化有关。     

p38MAPK表达与血管平滑肌细胞增殖关系的研究

目的探讨丝裂素活化蛋白激酶p38(p38MAPK)的表达与血管平滑肌细胞(VSMC)增殖的关系以及检测p38MAPK反义寡核苷酸(AODN)对VSMC增殖的抑制作用。方法将培养大鼠胸主动脉VSMC,随机分为对照组、p38MAPKAODN组、正义寡核苷酸(SODN)组。采用噻唑蓝比色分析法(MTT)和流式细胞仪检测VSMC,用蛋白免疫印迹法测定p38MAPK蛋白量。结果p38MAPKAODN能减少p38MAPK蛋白表达,明显抑制VSMC增殖,其抑制作用与p38MAPK蛋白表达相关,呈剂量依赖性。结论p38MAPKAODN能抑制大鼠VSMC增殖,该信号分子与VSMC增殖密切相关,可能是VSMC增殖的信号途径。     

腺病毒介导的p27kip1基因对胃癌细胞周期和DNA合成的影响

背景：近年来胃癌基因治疗的研究取得了一定的进展，但总体疗效尚不尽人意，目前正积极寻求组织特异性基因作为胃癌基因治疗的突破点。目的：以腺病毒为载体，研究p27kipl基因对胃癌细胞周期和DNA合成的影响。方法：将成功构建的携带人p27kipl基因的重组腺病毒载体Ad-p27kipl和LacZ重组腺病毒Ad-LacZ转染胃癌细胞系SGC-7901，并观察细胞形态的变化，流式细胞仪检测细胞周期和凋亡，^3H-胸腺嘧啶核苷(TdR)掺入实验测定细胞DNA合成。结果：Ad-p27kip1转染SGC-7901细胞后，细胞变圆、呈葡萄串样聚集以致脱落，G0／G1期细胞比例增加，8期、G2／M期细胞比例降低，并有凋亡发生，^3H-TdR掺入量亦显著降低。结论：腺病毒介导的p27kipl基因能使SGC-7901细胞产生G0／G1期阻滞，并能诱导细胞凋亡，抑制DNA合成，表明该基因疗法能有效抑制体外胃癌细胞的生长。     

一氧化碳对大鼠血管平滑肌细胞低氧性增殖反应的作用

目的 :从细胞周期的角度探讨降钙素基因相关肽 (CGRP)对人血管平滑肌细胞 (VSMC)增殖的影响 ,揭示其对抗动脉粥样硬化的作用机制。方法 :应用流式细胞仪分析CGRP对VSMC细胞周期的影响 ,免疫组织化学检测细胞周期蛋白D1 、E的表达。结果 :CGRP使细胞停留于G0 /G1 期 ,并能降低细胞中周期蛋白D1 、E的表达。结论 :CGRP可能是通过抑制周期蛋白D1 、E的积累 ,使人VSMC停留于G0 /G1 期 ,限制细胞周期的进程 ,抑制VSMC增殖     

卡托普利对血管平滑肌细胞增殖和PDGF—B、bFGF及相关癌基因的影响

采用内皮素建立培养的血管平滑肌细胞增殖模型。用氚-胸腺嘧啶核苷（［3H］－TdR）参入法，流式细胞术，免疫细胞化学及NorthemBlot杂交方法，观察卡托普利（Cap）对VSMC增殖的作用及对生长因子PDGF－B，bFGF及其相关癌基因c－sis，c－myc表达的影响。结果发现：Cap能逆转内皮素所致的［3H］－TdR参入量增多，阻止血管平滑肌细胞由静止期（G0／G1期）进入DNA合成期（S期）和有丝分裂（G2／M期），并能逆转内皮素引起的PDGF—B，bFGF抗原，c－sis，c－mycmRNA表达增强。提示：aop有抑制血管平滑肌细胞增殖作用，与生长因子及癌基因调控的分子生物学机制有关。     

粘着斑激酶反义寡核苷酸与胃癌细胞生长及凋亡的关系

根据FAKcDNA序列设计合成与FAKmRNA碱基序列互补的FAK反义寡核苷酸,导入BGC－823胃癌细胞,通过MTT比色法、细胞免疫化学染色法、逆转录－聚合酶链反应（RT－PCR）、细胞周期及电镜等方法,检测FAK反义寡核苷酸对人胃癌细胞的影响。结果示：①胃癌细胞生长明显受到抑制,抑制效应呈浓度和时间依赖性。②细胞内P125蛋白和FAKmRNA表达均明显下降。③经FAK反义寡核苷酸处理后BGC－823胃癌细胞发生凋亡,FCM普上可见明显的凋亡峰,电镜观察呈现凋亡早期形态改变,认为FAK反义寡核甘酸导入BGC－823胃癌细胞后,使BGC－823胃癌细胞FAKmPNA及P125蛋白表达水平下降,抑制PGC－823胃癌细胞增列,同时诱导BGC－823胃癌细胞发生凋亡。     

青少年高血压的研究进展

随着人们生活和行为方式的改变,高血压发病明显呈年轻化趋势。在青少年时期识别高血压病高危人群有助于早期进行有效干预和治疗,降低未来高血压的发生率及其严重性。现试从青少年高血压的诊断、发病因素、特点、治疗策略等方面的研究进展作一综述。     

Morbidity in cardiovascular diseases in immigrants in Sweden

INTRODUCTION: Although immigration to Sweden has increased in the last few decades, the incidence rates of cardiovascular disease and coronary heart disease in immigrants are unknown. The aim of the present study is to estimate whether place of birth affects the incidence rates of cardiovascular disease and coronary heart disease. MATERIAL AND METHODS: The study was designed as a follow-up study on morbidity in cardiovascular disease and coronary heart disease between 1 January 1997 and 31 December 1998, including three and a half million persons with age range 35-64 years, of whom 550 000 were born abroad, from the database MigMed consisting of the whole Swedish population. Incidence rates and relative risks were estimated by indirect standardization and a proportional hazard model. RESULTS: The age-adjusted risk of coronary heart disease was higher in most foreign-born groups than in Swedes. For example, in nine of 12 male groups, the relative risks varied between 1.1 and 2.2, and in seven of 12 female groups, the relative risks varied between 1.4 and 2.5. When also adjusting for level of education and employment status, the risks were still high, but on a lower level. CONCLUSIONS: Foreign-born people possess an over-risk of cardiovascular or coronary heart disease(CVD/CHD) compared with Swedish-born persons, also when level of education and employment status are taken into account.     

C·肉毒杀鼠素(冻干剂)应用研究

目的为研究C·肉毒杀鼠索对杀灭达乌尔黄鼠(简称黄鼠)的大面积应用情况和对家畜、家禽的毒害作用,进行了C·肉毒杀鼠素的应用研究.方法大面积投毒采用ES-2药饵撒播机[1],间隔约80m进行条投.羊、鸡采用直接灌胃.结果大面积应用的灭鼠率为83.72%.对羊、鸡最高剂量分别为500万MLD、150万MLD,均未出现中毒现象.结论 C·肉毒杀鼠素是较为理想的草原大面积杀灭黄鼠的理想、首选药物.     

高龄老年高血压的临床研究进展

随着社会老龄化进程的加快,80岁以上高龄老人的绝对数量以及占总人口的比例均在增加,如何提高高龄老年高血压的防治水平备受关注。现试从高龄老年高血压的临床特点、治疗策略等方面的临床研究进展作一综述。     

中国老年性耳聋的研究与干预进展

老年性耳聋已成为影响我国老年人生活质量的最主要的慢性病之一。助听器是目前帮助听力损失的老年人克服交流障碍的主要手段。在我国,数字助听器已逐渐取代模拟助听器并且体现出了更好的效果。但是老年听力损失患者中使用助听器的比例仍然很小。人工耳蜗植入也已被应用在老年患者中。我国针对老年人的听力康复服务还有较长的路要走。     

Sex Differences in Ethanol-induced Dopamine Release in Nucleus Accumbens and in Ethanol Consumption in Rats

In vivo microdialysis was used to examine changes in nucleus accumbens and striatal dopamine, dihydrophenylacetic acid (DO-PAC), and homovanillic acid (HVA) following acute administration of ethanol (0.0, 0.25, 0.5, 1.0, or 2.0 g/kg) in male and female Long-Evans rats. Following dialysis, rats were trained to bar-press for oral ethanol reinforcement. In nucleus accumbens, females showed significant increases in extracellular dopamine following 0.25 or 0.5 g/ kg ethanol, but did not show significant increases over baseline at the higher doses. Males showed slight increases in dopamine at the lower doses and decreased dopamine at 2.0 g/kg. In striatum, both sexes showed increased dopamine at the lower doses and decreased dopamine at 2.0 g/kg. There were slight increases in nucleus accumbens DOPAC and HVA at some doses in both sexes, but no changes in striatal metabolite levels. In addition to showing increased responsiveness to ethanol-induced mesolimbic dopamine stimulation, females consumed more ethanol than males during behavioral testing. The pattern of both greater ethanol-induced nucleus accumbens dopamine release and greater ethanol consumption in females supports the hypothesis that ethanol reward is mediated, at least in part, by the mesolimbic dopamine system.     

Epstein-Barr virus in malignancies in renal transplant recipients in Japan

A role for Epstein-Barr virus (EBV) in the development of malignancies including lymphomas, and carcinoma of the stomach, nasopharynx, thymus and salivary gland is suggested. It is indicated that EBV evokes polyclonal-B-cell-proliferative diseases in immunocompromised hosts, such as transplant patients, which results in monoclonal malignant lymphomas. The suppression of immune functions in these patients is thought to lead to incomplete elimination of the cells expressing EBV latent infection genes. To examine the etiological role of EBV in the development of malignancies following renal transplant in Japan, 42 malignancies in 1744 cases of renal transplant were studied for the presence and type of EBV. The polymerase chain reaction revealed that 5 malignancies were positive for EBV, all type A: 2 of 2 cases of non-Hodgkin's lymphoma (NHL), 2 of 8 cases of gastric adenocarcinoma of the common type, and 1 of 2 cases of gastric plasmacytoma. In situ hybridization revealed positive signals in the nucleus of tumor cells in 2 cases of NHL and 1 of plasmacytoma. Positive signals were found in the small lymphoid cells but not in the tumor cells in 2 cases of gastric carcinoma. On the basis of these findings, a role for EBV in the development of malignancies in renal transplant patients is unlikely except for lymphoid neoplasias.Abbreviations PCB polymerase chain reaction - EBV Epstein-Barr virus - NHL non-Hodgkin's lymphoma     

糖尿病大鼠心室肌细胞钾通道的改变

目的研究糖尿病大鼠心室肌细胞钾通道的改变及增加葡萄糖代谢对其的作用。方法取体重150～200g的雄性SpragueDawley大鼠,腹腔注射链脲菌素(STZ)建立糖尿病模型,采用酶解法获得单个心室肌细胞,应用膜片钳全细胞记录技术记录钾电流。结果糖尿病大鼠心室肌细胞瞬间外向性钾流(Ito)密度较对照组显著降低[ 60mV时,分别为(15.90±1.19)pA/pF(n=25)和(28.55±0.97)pA/pF(n=12),P<0.001];分别用100nmol/L胰岛素及1.5mmol/L二氯乙酸在体外预处理心室肌细胞4～5h和3～4h使Ito密度恢复至对照组水平[ 60mV时,分别为(29.40±0.38)pA/pF(n=20)和(27.35±0.97)pA/pF(n=12)]。结论糖尿病大鼠心室肌细胞钾通道功能发生改变,增加葡萄糖代谢可逆转这一改变,提示葡萄糖代谢与Ito功能间存在一定关系。