9型腺相关病毒转染乳鼠心肌细胞的可行性及安全性

背景：9型腺相关病毒对心肌细胞具有更好的靶向转染,是目前研究基因治疗心脏疾病的理想载体。目的：探索9型腺相关病毒体外转染乳鼠心肌细胞的可行性及对乳鼠心肌细胞的毒性评估。方法：分离培养原代乳鼠心肌细胞,以携带荧光蛋白基因的9型腺相关病毒为载体,将分离纯化的乳鼠心肌细胞分为正常培养组、无病毒转染组、病毒转染组。结果与结论：第1周细胞搏动频率及百分率正常培养组和病毒转染组与无病毒转染组比较,差异无显著性意义（P〉0.05）,但正常培养组高于病毒转染组（P〈0.05）;3周后各组比较,差异无显著性意义（P〉0.05）。经9型腺相关病毒转染的心肌细胞24h开始有表达,4~6d荧光最强,3组细胞72h内不同时间点还原比率均接近于1.0。说明9型腺相关病毒能成功有效地转染乳鼠心肌细胞,对乳鼠心肌细胞无明显毒性作用。     

鼠尾胶原联合血小板源性生长因子BB培养内皮细胞

背景:有研究证实,Ⅰ型鼠尾胶原可促进成肌纤维细胞的增加,对内皮细胞的迁移及成管有较为明显的作用,推断胶原可为细胞的生长提供一个较为合适的内环境。目的:探讨鼠尾胶原联合血小板源性生长因子BB抗人脐静脉内皮细胞凋亡的效果及安全性。方法:将第4代人脐静脉内皮细胞培养于铺有鼠尾胶原的培养皿上,用Alamar Blue法检测不同时间点的还原比率。将第4代人脐静脉内皮细胞分为4组培养于24孔板,其中生长因子组是在细胞接种前加入血小板源性生长因子BB蛋白于细胞悬浮液中;联合组需事先加入血小板源性生长因子BB蛋白于鼠尾胶中,铺于板底;最后各组均使用H2O2诱导凋亡,73 h后采用Western blot测定血小板源性生长因子BB、凋亡相关蛋白及抗凋亡蛋白的表达,同时Tunnel检测细胞凋亡阳性率。结果与结论:在铺有鼠尾胶原培养皿中培养人脐静脉内皮细胞的成管数量明显多于正常培养人脐静脉内皮细胞的成管数量(P<0.05)。鼠尾胶原培养的细胞与正常对照细胞生长状态相似,说明鼠尾胶原对细胞无明显毒性作用。联合组血小板源性生长因子BB、Bcl-2、p-Akt表达高于其余3组(P<0.05),Bax表达低于其余3组(P<0.05)。联合组细胞凋亡率低于生长因子组、H2O2组(P<0.05)。表明鼠尾胶原对人脐静脉内皮细胞无明显毒性作用,联合血小板源性生长因子BB生长因子后可显著增强抗细胞凋亡效果。     

前列腺环素合酶基因GLu461ALa多态性与维吾尔族人群心肌梗死的相关性

目的 探讨前列腺环素合酶基因(CYP8A1)GLu461ALa多态性与维吾尔族人群心肌梗死(myocardial infarction,MI)的相关性.方法 采用聚合酶链反应-限制性片段长度多态性方法,对210例维吾尔族MI患者和206名维吾尔族健康受试者CYP8A1基因4号外显子GLu461ALa位点进行分析,同时进行血清6-酮前列腺素F1α水平测定.结果 CYP8A1基因4号外显子GLu461ALa在MI组和健康对照组中基因璎频率分别为:CC型0.024(5/210)和0.010(2/206),AC型0.124(26/210)和0.073(15/206),AA型0.852(179/210)和0.917(189/206),两组CC、AC和从3种基因型频率分布差异无统计学意义(χ2=0.782,P>0.05),但CC+AC(突变纯合子+突变杂合子)在MI组[0.148(31/210)]明显高于对照组[0.083(17/206)],差异具有统计学意义(χ2=4.321,P=0.031),且在两组人群中C等位基因频率[0.086(36/420),0.046(19/412)]差异具有统计学意义(χ2=5.284,P=0.021).MI组血清6-酬前列腺素F1α水平[(17.40±4.56)pg/ml]低于对照组[(20.34±5.02)pg/ml],差异具有统计学意义(t=6.255,P<0.01);MI组和对照组CC+AC基因型携带者血清6-酮前列腺素Fα水平[(14.30±3.31)pg/ml,(18.31±4.62)pg/ml]均较AA基因型者[(19.34±5.51)pg/ml,(25.10±5.00)pg/ml]降低,差异具有统计学意义(t'=6.934,P<0.05;t=5.393,P<0.01).logistic回归分析显示,调整传统危险因素的干扰后,CYP8A1 C等位基因仍为MI的独立危险因素(OR=1.77;95%CI:1.06～2.05).结论CYP8A1基因4号外显子GLu461ALa多态性和维吾尔族人群MI的发生具有相关性,可能和基因变异导致的血清前列腺环素水平降低有关.     

纤维蛋白胶对脂肪干细胞增殖和存活的影响

背景:尽管有很多生物支架材料被用于携带种子细胞,但纤维蛋白胶与脂肪干细胞二者在体外的共培养研究及种子细胞在支架材料中的增殖存活评价没有统一的标准.目的:探讨纤维蛋白胶对体外培养的大鼠脂肪干细胞的增殖和存活性,为可注射组织工程化心肌治疗心肌梗死提供实验依据.方法:分离培养大鼠脂肪干细胞,通过流式细胞术检测其免疫表型,将脂肪干细胞与纤维蛋白胶共培养,用噻唑蓝比色法及LIVE/DEAD荧光双染色检测脂肪干细胞在纤维蛋白胶中的增殖和存活性.结果与结论:大鼠脂肪干细胞表型特征为CD90、CD105阳性,CD34、CD45阴性,噻唑蓝比色结果显示,纤维蛋白胶中的脂肪干细胞增殖活性高于常规培养组(P < 0.05),LIVE/DEAD荧光双染色显示脂肪干细胞在纤维蛋白胶中生长良好,24,72 h的细胞存活率均在90%以上,死亡率小于7%.结果提示,大鼠脂肪干细胞在纤维蛋白胶中增殖和存活良好,可用纤维蛋白支架携带脂肪干细胞用于可注射心肌组织工程研究.     

Association between matrix metalloproteinase-9 polymorphism (-1562C ＞ T/R279Q) and acute coronary syndrome in Uygur nationality of Xinjiang Autonomous Region of China

Objective To investigate the association between matrix metalloproteinase-9 (MMP-9) gene polymorphism (-1562C ＞ T/R279Q) and acute coronary syndrome (ACS) in Uygur nationality of Xinjiang Autonomous Region of China. Methods A total of 352 patients with ACS including 213 patients with unstable angina pectoris and 139 patients with acute myocardial infarction evidenced by using coronary arteriography and 421 control subjects were recruited in this study. The MMP-9-1562C ＞ T and R279Q genotypes were detemined by using PCR-RFLP method. The relationship between the polymorphism in the MMP-9 gene and the severity of coronary arterial stenosis was analyzed. All polymorphisms were determined for confimation with Hardy-Weinberg expectations in both groups separately. Differences in distributions of genotypes and alleles between two groups were analyzed with x2 test. The association between the MMP-9 polymorphisms and the risk of ACS was estimated by odds ratio(Ors) and their 95% confidence intervals (CIs), and the comprehensive evaluation of the factors associated with ACS was determined by using multifactor logistic regression. P ＜ 0. 05 was considered to be statistically significant. Results The genotype frequencies for CT + TT genotypes and T allele were 25.9 and14.5 percent in ACS subjects and 15.7 and 8.4 percent in control subjects, respectively. The genotype frequencies were different significantly between the two groups (x2 = 12.26,P ＜ 0.01;x2 = 14.15,P ＜ 0.01, respectively). No relationship between R279Q polymorphism and ACS was found in this study ( P ＞ 0.05). The multifactor logistic regression analysis showed that the T allele carrier (CT + TT) significantly increased the risk of ACS compared with the CC genotype ( OR = 1.791,95 % CI: 1. 088 - 2.951, P = 0.022) after adjustment for tradition risk factors. The frequencies for CT + TT and CC genotypes of the -1562C ＞ T polymorphism were not statistically different among ACS patients with one, two and three or more significantly diseased vessels ( x2 = 1.15, P = 0.56). Conclusions The findings suggest that the polymorphism in MMP-9 gene promoter (-1562C ＞ T) is associated with the susceptibility to the ACS. The T allele might be an independent risk factor for the ACS. But the -1562C ＞ T polymorphism may not be useful as a predictor of the severity of coronary arterial stenosis. The R279Q polymorphism of MMP-9 gene was not significantly associated with ACS in this studied population.     

老年人营养风险指数预测急性ST段抬高型心肌梗死患者经皮冠状动脉介入治疗的预后

目的旨在调查老年人营养风险指数(GNRI)是否与接受经皮冠状动脉介入治疗(PCI)急性ST段抬高型心肌梗死(STEMI)患者的死亡率相关。方法连续选取309例进行PCI的STEMI患者入组,将GNRI评分进行受试者工作特征曲线(ROC)分析,将GNRI≥94或94的患者分别被分配为0或1的GNRI评分组。结果在309例STEMI患者中,24例(7.74%)在医院死亡,15例(4.83%)在长期随访期间死亡[中位随访时间为19.5(3~36)月]。与GNRI 0组患者相比,GNRI 1组患者有更显著的在院死亡率(16.7%比4.4%,P0.001)和长期随访死亡率(23.8%比8.4%,P0.001)。GNRI(HR 2.039,95%CI 1.038~4.004,P=0.039)是接受PCI的STEMI患者死亡率的显著独立预测因子。此外,与GNRI 0组患者相比,GNRI 1组患者的累积生存率显著降低(76.2%比91.6%,log-rank P0.001)。结论 GNRI对于接受PCI的STEMI患者风险分层可能是有效的。     

9型腺相关病毒转染乳鼠心肌细胞的可行性及安全性**☆

背景：9型腺相关病毒对心肌细胞具有更好的靶向转染,是目前研究基因治疗心脏疾病的理想载体。 目的：探索9型腺相关病毒体外转染乳鼠心肌细胞的可行性及对乳鼠心肌细胞的毒性评估。 方法：分离培养原代乳鼠心肌细胞,以携带荧光蛋白基因的9型腺相关病毒为载体,将分离纯化的乳鼠心肌细胞分为正常培养组、无病毒转染组、病毒转染组。 结果与结论：第1周细胞搏动频率及百分率正常培养组和病毒转染组与无病毒转染组比较,差异无显著性意义(P > 0.05),但正常培养组高于病毒转染组(P < 0.05);3周后各组比较,差异无显著性意义(P > 0.05)。经9型腺相关病毒转染的心肌细胞24 h开始有表达,4~6 d荧光最强,3组细胞72 h内不同时间点还原比率均接近于1.0。说明9型腺相关病毒能成功有效地转染乳鼠心肌细胞,对乳鼠心肌细胞无明显毒性作用。     

血清淀粉样蛋白A1基因多态性对汉族人群颈动脉内中膜厚度的影响

Objective To explore the association between genetic polymorphism of serum amyloid protein A1 (SAA1) with carotid intima media thickness in a healthy Han Chinese population of Xinjiang. Methods A total of 449 healthy Han Chinese participating the cardiovascular risk survey between June 2007 and September 2009 were included, the genotypes of the SAA1 were detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). The mean IMT of the right and left common carotid arteries were measured by B-mode ultrasonography. Results (1) There was strong linkage disequilibrium between rs12218 and rs2229338 (D'=0. 89). (2) The carotid common IMT (CC-IMT) and the carotid bulb IMT (CB-IMT) were similar between the AA genotype (wild genotype) and the GGFAG genotype (mutational genotype) in rs2229338 of SAA1 gene. (3) CC-IMT[(0.081 ±0.071)cm vs (0.068 ±0. 019 ) cm, P = 0. 01] was significantly thicker in CC + CT genotype ( mutational genotype) group than in TT genotype (wild genotype) of rs12218 group and the difference remains significant after adjustment for age,gender,blood pressure, waist circumference, creatinine and high density lipoprotein cholesferoL CB-IMT [(0.085±0. 038)cm vs. (0.081 ± 0. 052) cm,P =0. 36] was similar between CC +CT genotype and TT genotype of rs12218 groups. Conclusion Our results suggested that the genetic polymorphism of SAA1 might be linked with IMT and rs12218 mutation could serve as a promoting factor for IMT in Han Chinese people.     

新疆汉、维吾尔、哈萨克族慢性心力衰竭流行病学调查及其患病率研究

目的 了解新疆成年人慢性心力衰竭(心衰)的患病率和分布特征.方法 应用四阶段整群随机抽样法,在全疆23个市、7个地区、5个自治州抽取乌鲁木齐市、克拉玛依市、阜康市、吐鲁番地区、和田地区、伊犁哈萨克自治州等6个地区年龄在35岁以上样本,男女均衡.统计不同民族组、不同年龄组、不同性别组人群的心衰患病率,并对心衰患者合并心血管基础疾病进行分析.结果 共抽样35岁以上城市及农村游牧居民8459例,心衰患病率为1.26%;其中汉族为0.89%,维吾尔族为1.11%,哈萨克族为2.14%.男性为1.61%,女性为0.93%,男性患病率高于女性(u=2.79,P<0.05).35～44岁、45～54岁、55～64岁、65～74岁、75岁以上各年龄组的心衰患病率分别为0.29%、0.60%、1.32%、2.55%、4.10%,随着年龄增高,心衰发生的风险显著上升.107例心衰患者中合并高血压病者占63.55%,冠心病者占42.99%,糖尿病者占18.69%,心瓣膜病者占5.61%,心房颤动者占4.67%.结论 新疆心衰患病率较高,随年龄增加呈增多趋势,且存在民族差异.高血压病、冠心病、糖尿病为新疆各民族心衰患者共同合并心血管基础疾病.通过对新疆不同人群心衰的流行病学监测和调查,寻找适合新疆人文地域特点的心衰早期诊断方法和干预模式具有重大现实意义.     

重组9型腺相关病毒载体转染小鼠心脏及对心功能的影响

目的 探讨重组9型腺相关病毒(recombinant adeno-associated serotype 9,rAAV9)载体对小鼠心脏的转染效果及对心功能的影响.方法 16只昆明种小鼠经尾静脉将携带增强型绿色荧光蛋白(enhanced green fluorescent protein,eGFP)报告基因的rAAV9(rAAV9-eGFP)转染入小鼠,于7、14、21、28 d留取标本,使用荧光显微镜观察eGFP在心肌、肝、肺、肾、脑、骨骼肌的荧光表达,Western blot法检测eGFP)的蛋白表达.另20只昆明种小鼠随机分为2组,每组10只,分别经尾静脉注射rAAV9-eGFP)和生理盐水,28 d后行心脏超声及血流动力学检查.结果 rAAV9-eGFP经尾静脉注射21 d时eGFP表达在心脏达到高峰,转染效率可达32%,在其他脏器表达量少或无表达.rAAV9-eGFP组较生理盐水组心功能差异无统计学意义(P＞0.05).结论 rAAV9可通过外周静脉注射在心脏较高表达,且对心功能无不良影响.