血管紧张素转换酶2基因多态性与原发性高血压合并脑卒中的关系

目的研究血管紧张素转换酶2（ACE2）基因G9570A多态性与我国南方汉族人原发性高血压合并脑卒中的关系。方法采用聚合酶链反应和限制性片段长度多态性（PCR-RFLP）的方法，检测156例原发性高血压患者，158例原发性高血压合并脑卒中患者及169例健康人群的ACE2基因，并进行组间对照研究，推测ACE2基因G9570A多态性与原发性高血压合并脑卒中发病的相关性。结果男性和女性原发性高血压组G等位基因频率分别为69．8％、57．1％，高于对照组的55．0％、44．2％，差异有统计学意义（P〈0．05）；两组ACE2基因型分布不同，原发性高血压组GG基因型的频率为32．8％，高于对照组的15．9％，差异有统计学意义（P〈0．05）。男性和女性原发性高血压合并脑卒中组A等位基因频率分别为57．3％、62．3％，高于原发性高血压组的30．2％、42．9％，差异有统计学意义（P〈0．01）；两组ACE2基因型分布不同，原发性高血压合并脑卒中组AA基因型的频率为37．7％，高于原发性高血压组的18．6％，差异有统计学意义（P〈0．01）。结论ACE2基因G9570A多态性与我国南方汉族人原发性高血压合并脑卒中可能具有一定相关性。携带A／AA基因的人群发生原发性高血压合并脑卒中的危险性相对较大。     

抗DNA酶B在急性肾小球肾炎诊断的临床应用

抗ＤＮＡ酶Ｂ在急性肾小球肾炎诊断的临床应用董太明黄震东符永恒吴红穗陈剑光陈志红罗健Ａ组溶血性链球菌（ＧＡＳ）感染导致的急性肾小球肾炎（ＡＧＮ）的病原学诊断，国内通常采用抗溶血素Ｏ（ＡＳＯ），到目前为止，尚未见到采用抗脱氧核糖核酸酶Ｂ（抗ＤＮＡ酶Ｂ）微...     

一氧化氮在幽门螺杆菌相关性胃病发病中的可能作用

目的探讨幽门螺杆菌(Hp)感染时胃粘膜内-氧化氮合酶(NOS)活性和一氧化氮(NO)含量与细胞凋亡改变的意义。方法选择慢性活动性胃炎病人27例,其中 Hp 阳性者15例,Hp 阴性者12例;正常对照组10例。应用生物化学方法和切口末端标记法(Tunnel)检测胃粘膜组织中一氧化氮产物 NO_2~-水平、NOS 活性及细胞调亡指数。结果 Hp 感染时,胃粘膜 NOS 活性、NO_2~-水平及细胞凋亡指数明显升高,较Hp 阴性组差异显著(P<0.01),胃炎积分数与胃粘膜组织中 NOS 活性和 NO_2~-水平呈明显正相关(γ=0.66和0.84,P<0.01)。当根除 Hp 后,胃粘膜组织 NOS 活性,NO_2~-水平及细胞凋亡指数则显著降低,细胞凋亡指数与 NOS 活性和 NO_2~-水平呈明显正相关(γ=0.68和0.79,P<0.01)。结论 Hp 感染时可引起胃粘膜组织 NOS 活化,NO 过量产生,细胞凋亡增加,这从又一方面说明 Hp 感染在胃腺癌发病机制中作用。     

Effect of carvedilol on attenuating the acute myocardial infarction-induced myocardial fibrosis in rats

Carvedilol,nonselective β-adrenoreceptor antagonist,was showed protective effects against acute myocardial infarction(AMI)-induced myocardial injury,however,the mechanisms underlying the antifibrosis effect of carvedilol has not been well known.The aim of the present study was to investigate the potential mechanism for the anti-fibrosis effect of carvedilol against AMI-induced myocardial fibrosis in rats.Methods Male SD rats were randomized into the sham group,LAD surgery-AMI model group,AMI plus low dose of carvedilol treatment group(1 mg /kg per day,CAR-L),AMI plus medium dose of carvedilol treatment group(5 mg /kg per day,CAR-M) and AMI plus high dose of carvedilol treatment group(10 mg /kg per day,CAR-H).The passage 3 neonatal SD rat cardiac fibroblasts were used for hypoxia /normoxia(2 h /4 h) treatment in the presence of carvedilol(0,1,2 and 4 μM).Results Cardiac remodeling and impaired heart function were observed after 14-week LAD surgery treatment,however,and the cardiac remodeling and decreased ejection fraction(EF%) and fractional shortening(FS%) were efficiently rescued in the CAR-M and CAR-H groups.The up-regulated expressions of Col1a1,Col3a1 and α-SMA at mRNA and protein levels were significantly reduced in the CAR-M and CAR-H groups.The in vitro study showed that Col1a1,Col3a1 and αSMA expressions at both mRNA and protein levels were down-regulated by carvedilol in rat cardiac fibroblasts in a dose-dependent manner.Smad3 inhibitors,SIS-3 and naringenin,could efficiently decrease Col1a1,Col3a1 and α-SMA expressions in rat cardiac fibroblasts.Smad3 was shown significantly inactivated in carvedilol-treated rat cardiac fibroblasts.Conclusion Carvedilol negatively regulates Smad3 signal pathway and inhibits extracellular matrix related Col1a1,Col3a1 and α-SMA expressions,contributing to the anti-fibrosis effect of carvedilol against AMI-induced myocardial fibrosis in rats.     

SPIO标记骨髓间充质干细胞移植入心肌梗死大鼠的示踪研究

目的探讨超顺磁性氧化铁微粒(SPIO)体外标记骨髓间充质干细胞(BMSCs)及体内示踪移植入大鼠心肌梗死心脏的BMSCs的能力。方法使用左旋多聚赖氨酸-SPIO共培养方式标记BMSCs。采用普鲁士蓝染色观察细胞内铁颗粒,流式细胞术检测细胞活力,将经过SPIO标记的干细胞移植入心肌梗死大鼠心脏,应用1.5TMRI系统行磁标记干细胞成像。超声心动图检测各组心脏的左室射血分数(EF),左室舒张末期内径(LVIDd),左室收缩末期内径(LVIDs)及短轴缩短率(FS)。结果普鲁士蓝染色显示,SPIO标记的BMSCs细胞胞质内出现细小的蓝色铁颗粒,标记效率为(99.81±1.57)%;与正常未标记细胞相比较,细胞的活力差异无统计学意义(P0.05)。标记了SPIO的BMSCs体内MRI成像时显示,细胞移植区域信号缺失,对应区域病理切片普鲁士蓝染色可见胞浆内染色阳性的细胞。超声心动图显示,PBS组FS移植前后没有明显变化,BMSC组FS从移植前的(23.1±1.88)%上升到第1周的(31.28±4.15)%。BMSC组EF在移植前是(51.13±5.07)%,第1周时上升到(60.12±8.40)%。结论 SPIO能成功地标记BMSCs,且对BMSCs的活力无明显影响。BMSCs移植后能改善心梗大鼠的心功能。SPIO标记的BMSCs移植后在大鼠体内的分布、迁移过程可用MRI进行检测评价。     

三七总皂苷对心梗后心室重构大鼠血流动力学作用

目的:研究三七总皂苷(PNS)对急性心肌梗塞(AMI)后左室重构(LVRM)大鼠血流动力学的影响。方法:结扎大鼠左冠状动脉前降支建立AMI模型,术后24 h随机分为对照组和实验组,连续4w分别灌胃给予生理盐水和PNS低、中、高剂量,观察PNS对病鼠心脏冠脉流量(CF)、左心室收缩压(LVSP)、左心室舒张末期压(LV-EDP)、左心室舒张压(LVDP)、等容收缩期左心室内压最大上升和下降速率(±dp/dtmax)、左心室收缩峰压(LVPP)、左室收缩速度(VPM)等心功能变化指标及心肌细胞病理结构和心脏指数改变的影响。结果:PNS可显著提高病鼠LVSP、CF、VPM及±dp/dtmax(P<0.01或P<0.05),显著降低LVEDP及LVPP(P<0.01或P<0.05),能显著改善病鼠左室心肌细胞形态结构的病理改变及心脏指数(P<0.01)。结论:PNS可通过扩张冠脉血管增加病鼠心脏冠脉流量,降低冠脉阻力,调整室壁顺应性,减轻心肌细胞结构病理损伤,明显改善左心收缩和舒张功能,对AMI后心脏血流动力学有改善作用。     

Lox-1基因501G＞C和3''''UTR*188C＞T多态性与急性心肌梗死发病的相关性

目的检测Lox-1501G〉C和3’UTR^＊188C〉T多态性在中国汉族人群中的分布及其与急性心肌梗死（acute myocardial infarction，AMI）发病的相关性。方法利用荧光染料SYBR Green Ⅰ标记定量PCR产物，通过PCR生长曲线和融解曲线分析结果进行多态位点的基因分型。分别检测汉族202名AMI患者及161名健康人群Lox-1 501G〉C和3’UTR^＊188C〉T多态性。随机选取30份经荧光定量PCR分型的标本进行DNA测序鉴定。结果AMI患者和健康人群的Lox-1 501G等位基因频率分别为0．834和0．798，Lox-1 501C等位基因频率分别为0．166和0．202，Lox-1 501G〉C基因型频率在AMI组和对照组问差异无统计学意义[OR0．687，95％CI（0．226—2．093）；P=-0．507]。AMI患者和健康人群的Lox-1 3’UTR^＊188C等位基因频率分别为0．681和0．674，Lox-1 3’UTR^＊188T等位基因频率分别为0．319和0．326．Lox-1 3’UTR^＊188C〉T基因型频率在AMI组和对照组亦无统计学意义[OR 0．947，95％CI（0．623-1．439）；P=0．80]。结论中国汉族Lox-1基因501G〉C和3’UTR^＊188C〉T多态性与AMI发病无关。     

人嗜铬粒蛋白A N-端片段Vasostatin-1(CGA1-76)的原核表达

目的制备编码人Vasostatin-1基因片段,利用大肠杆菌表达重组Vasostatin-1蛋白。方法根据人Vasostatin-1基因序列,设计、合成3对PCR引物,利用PCR合成法制备编码Vasostatin-1的DNA。将Vasostatin-1基因定向插入原核表达载体pGEX-4T-1,行酶切和DNA测序鉴定,并将构建的重组质粒转化大肠杆菌BL21(DE3)。用IPTG诱导BL21(DE3)中导入的Vasostatin-1基因的表达,行SDS-PAGE分析。用GSTrap亲合柱纯化重组Vasostatin-1蛋白。结果用PCR合成法制备了228bp的Vasostatin-1基因片段,并成功构建了重组质粒pGEX-4T-Vasostatin-1,DNA测序显示Vasostatin-1DNA序列和插入位点正确。经IPTG诱导,特异表达出以包涵体形式存在36kDa的重组Vasostatin-1蛋白。结论制备、克隆了人Vasostatin-1基因片段,并在大肠杆菌中表达出重组Vasostatin-1蛋白。     

Screen for Coronary Artery Disease Specific Genetic Expression by Representational Differential Analysis

Objective To screen coronaryartery disease (CAD) specific expressions and clone their genes. Method Blood samples were collected from CAD and non - CAD patients at the end of coronary angiography. mRNA from samples was isolated and converted into cDNA. After ligated with specific linkers, the cDNA was amplified with complementary primers. PCR products from CAD samples were named as tester; the ones from non - CAD samples were named as driver. With different ratio of tester to driver (1 : 100,1: 1, 000, and 1: 10, 000), they were mixed, denatured, and renatured. Single strand cD-NA was eliminated by Mung bean nuclease. Double strand cDNA presented only in tester was amplified, ligated in vector pUC19 and pUC53, and transformed into E. coll DH5a. Strains with inserted cDNA fragments were picked up based on blue and white selection. Insertions were screened by endonuclease digestion and DNA sequencing. Results were compared with DNA sequences of GeneBank. Results: After the selection with representational