黄芪、当归对血管内皮剥脱后内膜增生的影响及作用机制

目的 研究黄芪和当归防治血管内皮剥脱后再狭窄与血管平滑肌细胞(VSMC)表型转化之问的关系。方法 建立大鼠主动脉内皮剥脱模型，分别采用形态学和Northern印迹分析技术，观察大鼠血管内皮剥脱后内膜增生情况及VSMC表型标志基因SM α—肌动蛋白、平滑肌胚胎型肌聋蛋白重链(SMemb)表达活性。结果 术后7d模型组内膜明显增生，14～21d内膜呈进行性弥漫性增厚，与模型组相比，两种中药治疗组的内膜增生程度均明显藏轻1分化标志SM α—肌动蛋白基因表达增高，去分化标志SMemb表达降低。结论 黄芪和当归通过抑制VSMC表型转化而减缓血管内膜增生。     

反义c-jun表达质粒对血管平滑肌细胞表型转化标志的影响

为研究c-jun反义RNA对血管平滑肌细胞表型转化的影响,本文应用可表达c-jun反义RNA的真核细胞表达载体转染大鼠血管平滑肌细胞,采用Northern印迹、Westem印迹和氚标胸腺嘧啶核苷掺入实验,观察反义c-jun对血管平滑肌细胞表型标志基因平滑肌α-肌动蛋白、平滑肌胚胎型肌球蛋白重链和骨桥蛋白表达和DNA合成的影响。结果发现,在血管平滑肌细胞中表达的c-jun反义RNA可使血管平滑肌细胞去发化标志基因平滑肌胚胎型肌球蛋白重链和骨桥蛋白mRNA表达较对照分别下降50％和95％,骨桥蛋白水平较对照下降75％,同时血管平滑肌细胞的DNA合成受到显著抑制,实验组氚标胸腺嘧啶核苷掺入值对照降低37％。c-jun反义RNA对血管平滑肌细胞分化标志基因平滑肌α－肌动蛋白的表达无明显影响。提示c-jun基因表达产物在维持血管平滑肌细胞于去分化状态中起重要作用。     

碱性成纤维细胞生长因子反义寡脱氧核苷酸抑制胰岛素诱导动脉平滑肌细胞增殖

为研究胰岛素诱导的动脉平滑肌细胞增殖作用和碱性成纤维细胞生长因子反义寡脱氧核苷对培养的Ｗｉｓｔａｒ大鼠主动脉平滑肌细胞生长的影响。采用胰岛素和碱性成纤维细胞生长因子反义寡脱氧核苷酸处理培养的Ｗｉｓｔａｒ大鼠主动脉平滑肌细胞,Ｎｏｒｔｈｅｎｂｌｏｔ检测碱性成纤维细胞生长因子基因ｍＲＮＡ表达,并测定氚标胸腺嘧啶脱氧核苷掺入和细胞计数。结果发现胰岛素能明显诱导平滑肌细胞碱性成纤维细胞生长因子ｍＲＮＡ表达和增殖,且呈浓度依赖性。胰岛素浓度由０．００１ｍｇ／Ｌ增加到１．０ｍｇ／Ｌ,平滑肌细胞增殖率由１０％增加到５３％。碱性成纤维细胞生长因子反义寡脱氧核苷酸（５μｍｏｌ／Ｌ）能明显抑制胰岛素诱导的平滑肌细胞碱性成纤维细胞生长因子ｍＲＮＡ表达和增殖,氚标胸腺嘧啶脱氧核苷掺入被抑制４１．１％（与对照组比较,Ｐ＜０．０１）,细胞计数被抑制３０．２％（与对照组比较,Ｐ＜０．０１）。提示胰岛素可通过诱导平滑肌细胞碱性成纤维细胞生长因子的表达而促进动脉平滑肌细胞增殖,碱性成纤维细胞生长因子反义寡脱氧核苷酸能有效抑制胰岛素诱导的平滑肌细胞的ＤＮＡ合成及其增殖。     

新生小牛血清,碱性成纤维细胞生长因子和肝素对大鼠血管平滑肌细胞 …

为探讨新生小牛血清和碱性成纤维细胞生长因子促血管平滑肌细胞迁移与肝素抑制新生小因清促血管平滑肌细胞迁移的分子机制,采用反转录聚合酶链反应方法合成骨桥蛋白ｃＤＮＡ探针,以羟基脲抑制新生小牛血清、碱性成纤维细胞生长因子对血管平滑肌细胞的促增殖作用后,观察其对骨桥蛋白基因表达及能量转换的影响。结果发现,新生小牛血清、碱性成纤维细胞生长因子均能显著诱导血管平滑肌细胞骨桥蛋白基因表达,在两种因素作用下肌酸激     

碱性成纤维细胞生长因子在烟草烟雾诱导大鼠血管平滑肌细胞增殖中的作用

目的探讨烟草烟雾提取物对大鼠血管平滑肌细胞增殖的影响及碱性成纤维细胞生长因子在其中的作用。方法按不同浓度烟草烟雾提取物(0、2.5%、5%、10%和20%)分为对照组、低浓度、中等浓度、高浓度和过高浓度烟草烟雾提取物组刺激血管平滑肌细胞,采用MTT法观察细胞增殖变化,免疫细胞化学法测定碱性成纤维细胞生长因子和增殖细胞核抗原蛋白的表达,同时用逆转录-聚合酶链反应法检测碱性成纤维细胞生长因子mRNA表达。用筛选出的最适烟草烟雾提取物浓度处理大鼠血管平滑肌细胞不同时间(0、4、8、12 h和24 h)后,检测碱性成纤维细胞生长因子mRNA及碱性成纤维细胞生长因子和增殖细胞核抗原蛋白的变化。用碱性成纤维细胞生长因子抗体和最适浓度烟草烟雾提取物干预血管平滑肌细胞24 h后检测细胞增殖及碱性成纤维细胞生长因子和增殖细胞核抗原蛋白表达的变化。结果 (1)与对照组相比,低浓度烟草烟雾提取物组(P0.05)和中浓度组血管平滑肌细胞增加明显(P0.01),而高浓度组和过高浓度组与对照组比较差异无显著性(P0.05)。碱性成纤维细胞生长因子mRNA、蛋白和增殖细胞核抗原蛋白在对照组中有少量表达,低浓度烟草烟雾提取物组表达增加(P0.01),中浓度烟草烟雾提取物组达到高峰,高浓度和过高浓度烟草烟雾提取物组仍高于对照组(P0.01)。(2)对照组(不加烟草烟雾提取物组即0 h组)血管平滑肌细胞中有少量碱性成纤维细胞生长因子mRNA、蛋白和增殖细胞核抗原蛋白表达。低浓度烟草烟雾提取物刺激4 h后细胞内碱性成纤维细胞生长因子mRNA、蛋白和增殖细胞核抗原蛋白表达增加(P0.01),碱性成纤维细胞生长因子mRNA于8 h达高峰;碱性成纤维细胞生长因子和增殖细胞核抗原蛋白于12 h达高峰。(3)碱性成纤维细胞生长因子抗体可显著抑制5%烟草烟雾提取物诱导的血管平滑肌细胞增殖和碱性成纤维细胞生长因子、增殖细胞核抗原蛋白表达增加。结论低浓度和中浓度烟草烟雾提取物对大鼠血管平滑肌细胞的促增殖作用逐渐增加;高浓度和过高浓度组时促增殖作用反而减弱。烟草烟雾提取物可能是通过增加碱性成纤维细胞生长因子的表达促进大鼠血管平滑肌细胞的增殖。     

新生小牛血清、碱性成纤维细胞生长因子和肝素对大鼠血管平滑肌细胞骨桥蛋白基因表达的影响

为探讨新生小牛血清和碱性成纤维细胞生长因子促血管平滑肌细胞迁移与肝素抑制新生小牛血清促血管平滑肌细胞迁移的分子机制,采用反转录聚合酶链反应方法合成骨桥蛋白ｃＤＮＡ探针,以羟基脲抑制新生小牛血清、碱性成纤维细胞生长因子对血管平滑肌细胞的促增殖作用后,观察其对骨桥蛋白基因表达及能量转换的影响。结果发现,新生小牛血清、碱性成纤维细胞生长因子均能显著诱导血管平滑肌细胞骨桥蛋白基因表达,在两种因素作用下肌酸激酶活性分别较对照组升高７６％和６１％,具有显著性差异（Ｐ＜０．０５）;在肝素与新生小牛血清共同作用下,骨桥蛋白基因表达减弱,细胞肌酸激酶活性也较新生小牛血清单独作用时降低２２％。提示新生小牛血清和碱性成纤维细胞生长因子促进血管平滑肌细胞迁移以及肝素抑制新生小牛血清促血管平滑肌细胞迁移的作用与细胞内骨桥蛋白基因表达改变和能量转换有关。     

碱性成纤维细胞生长因子反义寡脱氧核苷酸抑制胰岛素诱导动 …

为研究胰岛素诱导的动脉平滑肌细胞增殖作用和碱性成纤维细胞生长因子反义寡脱氧核苷对培养的Ｗｉｓｔａｒ大鼠主动脉平滑肌细胞生长的影响。采用胰岛素和碱性成纤维细胞生长因子反义寡脱氧核苷酸处理培养的Ｗｉｓｔａｒ大鼠主动脉平滑肌细胞,Ｎｏｒｔｈｅｎｂｌｏｔ检测碱性成纤维细胞生长因子基因ｍＲＮＡ表达,并测定氚标胸腺嘧啶脱氧核苷掺入和细胞计数。结果发现胰岛素能明显诱导平滑肌细胞碱性成纤维细胞生长因子ｍＲＮＡ表     

肝细胞生长因子及其受体在血管外膜成纤维细胞中的表达

目的 检测肝细胞生长因子（HGF）及其受体c-met在血管外膜2种表型成纤维细胞中的表达，探索其在血管外膜重塑中的作用。方法 用转化生长因子-β1建立大鼠主动脉外膜成纤维细胞和肌成纤维细胞2种细胞表型。以实时定量逆转录聚合酶链反应技术和蛋白免疫印迹方法分析HGF及c-met在2种不同表型细胞中的表达。酶联免疫吸附试验检测2种细胞无血清培养上清液中HGF的含量。结果 HGFmRNA及HGF蛋白，c-metmRNA及c-met蛋白在主动脉外膜成纤维细胞。肌成纤维细胞中均有表达。在主动脉外膜成纤维细胞转变为肌成纤维细胞后表达明显上调，随转化生长因子-β1刺激浓度和时间的增加，在主动脉外膜成纤维细胞逐渐表达-α-平滑肌肌动蛋白转变为肌成纤维细胞的过程中HGF表达逐渐上调。结论 血管外膜成纤维细胞存在局部HGF系统。HGF可能在血管外膜细胞瑶型转化中有一定作用。     

罗格列酮抑制体外培养大鼠血管平滑肌细胞的增殖和迁移作用

目的 探讨罗格列酮对大鼠主动脉平滑肌细胞增殖及迁移行为的影响.方法 组织贴块法原代培养大鼠主动脉平滑肌细胞,胞浆内免疫组织化学染色鉴定细胞.取第5代纯化细胞无血清培养使细胞同步,用血小板源性生长因子BB或碱性纤维细胞生长因子诱导细胞增殖和迁移,之前不加或加入罗格列酮(1、5和10μmol/L)进行预处理.应用免疫组织化学染色法检测细胞核内增殖细胞核抗原的表达,流式细胞仪分析细胞周期分布,Boyden趋化小室观测细胞迁移能力.结果 罗格列酮能够显著抑制血小板源性生长因子BB或碱性纤维细胞生长因子诱导的平滑肌细胞核内增殖细胞核抗原的表达,抑制细胞由G0/G1期向S期的转变,抑制细胞在Boyden室间的迁徙,且呈明显的荆量依赖关系(P<0.0001).结论 罗格列酮能够抑制血小板源性生长因子BB或碱性纤维细胞生长因子诱导的血管平滑肌细胞的增殖和迁移.     

碱性成纤维细胞成长因子、新生小牛血清和肝素对大鼠血管平滑肌细胞迁移的影响

为研究血管平滑肌细胞迁移及其影响因素，建立体外检测血管平滑肌细胞迁移的实验方法，并观察不同浓度的新生小牛血清、碱性成纤维细胞生长因子和肝素作用于血管平滑肌细胞不同时间后对其迁移的影响。结果发现，新生小牛血清和碱性成纤维细胞生长因子均可显著促进血管平滑肌细胞迁移（P＜0.001或P＜0.01）；肝素对新生小牛血清诱导的血管平滑肌细胞迁移产生显著抑制作用（P＜0．001）．三者的作用强度均与浓度呈正相关。提示碱性成纤维细胞生长因子对血管平滑肌细胞具有促增殖作用．而肝素具有抑制血管平滑肌细胞迁移的作用。     

妊娠时间与肺结核复发的相关性研究及诊治对策

目的 分析肺结核史患者妊娠时间和肺结核复发间相关性.方法 选取我院收治的有肺结核史的妊娠妇女576例作为研究对象,对其妊娠前肺结核治疗、治愈后妊娠时间、妊娠后复发肺结核等进行分析,总结有肺结核史育龄女性的妊娠时间和肺结核复发之间的关系.结果 肺结核治愈后不同时间段妊娠者的结核复发率比较,差异具有显著性（P＜0.05）,停药后间隔时间越久妊娠,肺结核复发的几率越小.结论 加强孕期痰菌检查,及早发现复发肺结核,提高母婴安全.     

我国骨关节结核诊治的回顾与展望

骨关节结核是危害人们健康的严重感染性疾病,近95％由他处结核病继发而来.罹患骨关节结核疾病后几乎均将致残,严重影响人们的健康、工作和生活.建国以来在党和国家的关心和支持下,骨关节结核的诊治水平取得了长足进步.时至今日,由于多种原因,学科发展和被重视程度受到一定的制约,同整个医疗行业的发展不相适应.回顾过去,展望未来,我们需要重新审视骨关节结核的诊治方法,努力推进骨关节结核诊疗技术的科学发展.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fos and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44~(MAPK), p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44~(MAPK), p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44~(MAPK) and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between P42/44~(MAPK) and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Raf/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44~(MAPK), c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

Overweight and Obesity and Progression of ADPKD

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Effect of phosphorylation of MAPK and Stat3 and expression of c-fas and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44MAPK, p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44MAPK, p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44MAPK and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between p42/44MAPK and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Rat/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44MAPK, c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice

Non-invasive techniques to monitor stress hormones in small animals like mice offer several advantages and are highly demanded in laboratory as well as in field research. Since knowledge about the species-specific metabolism and excretion of glucocorticoids is essential to develop such a technique, we conducted radiometabolism studies in mice (Mus musculus f. domesticus, strain C57BL/6J). Each mouse was injected intraperitoneally with 740 kBq of 3H-labelled corticosterone and all voided urine and fecal samples were collected for five days. In a first experiment 16 animals (eight of each sex) received the injection at 9 a.m., while eight mice (four of each sex) were injected at 9 p.m. in a second experiment. In both experiments radioactive metabolites were recovered predominantly in the feces, although males excreted significantly higher proportions via the feces (about 73%) than females (about 53%). Peak radioactivity in the urine was detected within about 2h after injection, while in the feces peak concentrations were observed later (depending on the time of injection: about 10h postinjection in experiment 1 and about 4h postinjection in experiment 2, thus proving an effect of the time of day). The number and relative abundance of fecal [3H]corticosterone metabolites was determined by high performance liquid chromatography (HPLC). The HPLC separations revealed that corticosterone was extensively metabolized mainly to more polar substances. Regarding the types of metabolites formed, significant differences were found between males and females, but not between the experiments. Additionally, the immunoreactivity of these metabolites was assessed by screening the HPLC fractions with four enzyme immunoassays (EIA). However, only a newly established EIA for 5alpha-pregnane-3beta,11beta,21-triol-20-one (measuring corticosterone metabolites with a 5alpha-3beta,11beta-diol structure) detected several peaks of radioactive metabolites with high intensity in both sexes, while the other EIAs showed only minor immunoreactivity. Thus, our study for the first time provides substantial information about metabolism and excretion of corticosterone in urine and feces of mice and is the first demonstrating a significant impact of the animals' sex and the time of day. Based on these data it should be possible to monitor adrenocortical activity non-invasively in this species by measuring fecal corticosterone metabolites with the newly developed EIA. Since mice are extensively used in research world-wide, this could open new perspectives in various fields from ecology to behavioral endocrinology.     

Replication and Inhibitors of Enteroviruses and Parechoviruses

The Enterovirus (EV) and Parechovirus genera of the picornavirus family include many important human pathogens, including poliovirus, rhinovirus, EV-A71, EV-D68, and human parechoviruses (HPeV). They cause a wide variety of diseases, ranging from a simple common cold to life-threatening diseases such as encephalitis and myocarditis. At the moment, no antiviral therapy is available against these viruses and it is not feasible to develop vaccines against all EVs and HPeVs due to the great number of serotypes. Therefore, a lot of effort is being invested in the development of antiviral drugs. Both viral proteins and host proteins essential for virus replication can be used as targets for virus inhibitors. As such, a good understanding of the complex process of virus replication is pivotal in the design of antiviral strategies goes hand in hand with a good understanding of the complex process of virus replication. In this review, we will give an overview of the current state of knowledge of EV and HPeV replication and how this can be inhibited by small-molecule inhibitors.     

荣宝和氯硝柳胺灭螺效果比较及成本分析

目的 评价新型灭螺药物荣宝杀灭钉螺的效果,探讨其推广应用价值.方法 按目前推荐的荣宝灭螺剂量,喷洒法为30 g/m2,浸杀法为50 g/m3;氯硝柳胺喷洒法和浸杀法分别采用2 g/m2和2 g/m3杀螺剂量,分别在室内和现场进行灭螺试验,观察两种药物的灭螺效果并初步分析评估其成本.结果 在现场气温22～30℃条件下,荣宝50 g/m3浸杀3、5、7 d后,螺袋内钉螺校正死亡率均达到100.0%,与氯硝柳胺2 g/m3灭螺效果相似;荣宝30 g/m2剂量喷洒3、5、7、15 d后,钉螺校正死亡率分别为54.5%、58.0%、69.0%、79.1%,氯硝柳胺喷洒组钉螺校正死亡率分别为61.0%、69.4%、76.7%、77.9%.在室温18℃条件下,荣宝以30 g/m2喷洒3、5、7、15 d后,钉螺校正死亡率分别为72.9%、87.2%、91.5%、76.1%;而相应2 g/m2氯硝柳胺喷洒后的钉螺校正死亡率分别为81.3%、95.7%、97.9%、80.4%.同样完成1000 m2的喷洒灭螺任务,荣宝所需灭螺药物和人力资费成本比氯硝柳胺多支出0.114元/m2;完成72 m3的浸杀灭螺任务,荣宝所需灭螺药物和人力资费成本比氯硝柳胺多支出0.127元/m3.50 g/m3荣宝浸杀灭螺剂量,对成鱼(＞250 g)的活力不会造成影响,但对鱼类幼苗仍具较强毒性.结论 荣宝与氯硝柳胺灭螺效果相似,由于其成本较高,氯硝柳胺仍然是目前首选灭螺药物,但荣宝的鱼类毒性低,可作为氯硝柳胺之外有益的补充灭螺药物.     

心电图、心电向量图与冠状动脉造影结果对比分析

目的:通过分析心电图(Electrocardiogram,ECG)和心电向量图(Vectorcardiogram,VCG)的改变与冠脉造影（CAG）结果进行对比,探讨ECG、VCG在冠状动脉病变中的诊断价值。方法: 选择2008年1月~2009年12月临床拟诊断为冠心病患者108例,行常规ECG、VCG检查,并于1周内进行CAG,对检查结果依据各自的诊断标准进行判定,以CAG为标准诊断法,利用四格表法,计算相关评价真实性的指标并进行比较。结果: ①VCG检测的灵敏度、特异度、准确度显著高于ECG(P<0.05,P<0．01)。②ECG、VCG阳性率与冠脉病变支数组间比较:在单支病变、双支病变中,VCG阳性率明显高于ECG(P<0.05),左主干或三支病变无统计学意义;组内比较:ECG组左主干或三支病变组较单支病变、双支病变阳性率高(P<0.05,P<0.01);VCG组左主干或三支病变组较单支病变阳性率高(P<0.05);与双支病变阳性率比较无统计学意义;③ECG、VCG阳性率与冠脉病变程度组间比较:冠脉病变狭窄50%～69%的VCG阳性率明显高于ECG (P<0.05),其他两组阳性率比较无统计学意义;组内比较:ECG组冠脉病变狭窄≥90%较50%～69%、70%～89%的阳性率高(P<0.05,P<0.01); VCG组狭窄≥90%较50%～69%阳性率高（P<0.01),其他无统计学意义。结论: VCG对冠心病检测价值显著高于ECG。     

Detection and characterization of the product of hydroethidine and intracellular superoxide by HPLC and limitations of fluorescence

Here we report the structural characterization of the product formed from the reaction between hydroethidine (HE) and superoxide (O(2)(.-)). By using mass spectral and NMR techniques, the chemical structure of this product was determined as 2-hydroxyethidium (2-OH-E(+)). By using an authentic standard, we developed an HPLC approach to detect and quantitate the reaction product of HE and O(2)(.-) formed in bovine aortic endothelial cells after treatment with menadione or antimycin A to induce intracellular reactive oxygen species. Concomitantly, we used a spin trap, 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO), to detect and identify the structure of reactive oxygen species formed. BMPO trapped the O(2)(.-) that formed extracellularly and was detected as the BMPO-OH adduct during use of the EPR technique. BMPO, being cell-permeable, inhibited the intracellular formation of 2-OH-E(+). However, the intracellular BMPO spin adduct was not detected. The definitive characterization of the reaction product of O(2)(.-) with HE described here forms the basis of an unambiguous assay for intracellular detection and quantitation of O(2)(.-). Analysis of the fluorescence characteristics of ethidium (E(+)) and 2-OH-E(+) strongly suggests that the currently available fluorescence methodology is not suitable for quantitating intracellular O(2)(.-). We conclude that the HPLC/fluorescence assay using HE as a probe is more suitable [corrected] for detecting intracellular O(2)(.-).