家族性载脂蛋白B-100缺陷症的检测

目的：建立检测家族性载脂蛋白Ｂ－１００缺陷症（ＦＤＢ）的方法和研究ＦＤＢ与高脂血症和冠心病的关系。方法：用聚合酶链反应扩增载脂蛋白Ｂ－１００基因特定片段，以非放射性物质地高辛标记等位基因特异性寡核苷酸作为探针与扩增产物杂交，检测突变载脂蛋白Ｂ基因。结果：在１０３例高脂血症患者中发现１例突变，１００例冠心病患者中有２例突变，３４５例正常人中无突变者。结论：该基因与脂质代谢和冠心病有关；本试验方法简便、稳定，适用于临床ＦＤＢ的普查     

汉族人 CST3单核苷酸多态性与冠心病、胱抑素 C水平相关性研究

目的：研究胱抑素 C 基因（CST3）－157位点单核苷酸多态性（SNP）与冠心病及血胱抑素 C（Cys C）水平的相关性。方法：采用病例对照研究,入选因疑似冠心病入院的282例患者为研究对象,依据冠脉造影结果分为冠心病组（n＝130）及对照组（n＝152）。提取外周血白细胞基因组 DNA,应用巢式 PCR-直接测序技术检测 CST3－157位点 SNP,分析该位点的基因型分布及等位基因频率与冠心病、Cys C 水平的关系。结果：汉族人群中 CST3基因－157位存在 G／C 突变,存在 GG、GC 和 CC 3种基因型, GG 型为野生基因型,无论等位基因频率或者基因型分布在冠心病及对照组无明显差异,但野生基因型 Cys C 水平明显高于突变基因型。结论：汉族人群中 CST3基因－157位基因多态性与冠心病发病无直接相关性,携带野生型者血 Cys C 水平较高。     

用PCR和直接测序方法分析胰腺的c-Ki-ras基因点突变

用ＰＣＲ和直接测序方法分析胰腺的ｃ－Ｋｉ－ｒａｓ基因点突变徐永泉，刘述信，夏玉亭人类肿瘤中最常见的与转化活性有关的基因ｒａｓ家族癌基因（Ｎ－ｒａｓ，Ｋｉ－ｒａｓ，Ｈａ－ｒａｓ）。ｒａｓ基因的激活在肿瘤发生中起重要作用。ｒａｓ基因激活的主要方式是点突变...     

高胆固醇血症患者筛查出家族性载脂蛋白B-100缺陷症

目的：筛查高胆固醇血症患载脂蛋白（ａｐｏ）Ｂ－１００可能存在的突变体。方法：以聚合酶链反应结合单链构象多态性分析筛查受检ｓｐｏ Ｂ基因３５００密码子附近一段ＤＮＡ是否存在突变，对产生异常图谱的ＤＮＡ片段进行测序。先证的基因型以ａｐｏＢ基因的三个多态点定出。结果：在３６２例高胆固醇血症患中，发现１例Ｒ３５００Ｗ先证，基因型为ＸｂａⅠ－／－，ＭｓｐⅠ＋／＋，ＥｃｏＲⅠ＋／＋。结论：广州人群存在ａｐｏＢ－１００ Ｒ３５００Ｗ突变携带，与携带同一突变的台湾及海外华人可能来自同一远祖。     

ras癌基因第12密码子点突变和胃癌患者预后关系的研究

对福尔马林固定，石蜡包埋的胃癌组织使用移聚酶链式反应－限制性片段长度多态性（简称ＰＣＲ－ＲＦＬＰ）技术同时进行ｃ－Ｈａ－ｒａｓ基因第１２位和６１位，Ｎ－ｒａｓ基因第１２位，ｋ－ｒａｓ第１２位和１３位密码子点突变的研究，发现３３．３％（１４／４２），的胃癌有ｃ－Ｈａ－ｒａｓ基因第１２位密码子的点突变；４．８％（２／４２）的病例有ｋ－ｒａｓ基因第１２位密码子的点突变，点突变的发生与患者的预后，淋巴结转     

编码肝片吸虫GST和猪蛔虫GST基因的测序及同源性分析

目的：比较肝片吸虫GST（FhGST）、猪蛔虫GST（AsGST）基因与日本血吸虫26GST（Sj26GST）间的同源性。方法：以盐酸胍／氯化铯梯度超速离心法提取肝片吸虫和猪蛔虫的总RNA，合成两对特异性引物，通过反转录－PCR将所需的编码GST的cDNA扩增出来，并将PCR产物直接测序。应用DNASIS软件比较FhGST、AsGST基因与Sj26GST间的同源性。结果：测序得到FhGST403bp片段和AsGST411bp片段。结论：FhGST和AsGST片段与 Sj26GST的cD－NA在核苷酸水平上的同源性分别为59％和54％，两片段间的同源性为52％。     

临床分离耐氟喹诺酮金黄色区区球菌的耐药机制研究

为了解临床分离金黄色葡萄球菌（金葡萄）对氟喹诺酮类药物的耐药机制。应用聚合酶链反应－限制性片段长度多生（ＰＣＲ－ＲＦＬＰ）分析和ＰＣＲ－边构象多太分析（ＳＳＣＰ）技术及利血平逆转实验，分别检测３１株临床分离耐环丙沙星金谋略 株ｇｙｒＡ基因的突变及ｎｏｒＡ基因的表达。结果表明，３１菌中２２株检出ｇｒｙＡ基因８４位点突变，有２８株在得血平逆转实验中对环丙沙星和诺氟沙星的ＭＩＣ同时降低，表明存在 ｏｒＡ     

线粒体tRNAGuluA14693G突变与2型糖尿病的易感性研究

目的 研究湖北地区2型糖尿病（T2DM）中线粒体基因tRNAGlu A14693G和tRNALeu（UCR）A3243G突变的发生率及其与T2DM相关性。方法 采用聚合酶链反应-限制性片段长度多态性（PCR—RFLP）结合基因测序技术，对179例T2DM患者和208例糖耐量正常的健康对照进行检测。结果首次在T2DM中发现4例tRNAGlu（A14693G）突变（2．23％），对照组未检出该突变，两组间A14693G突变率差异有统计学意义（X^2=4．697．P=0．030）；未发现A3243G的点突变。结论 tRNA^GluA14693G突变可能与T2DM的易感性有关。     

男性不育患者不育相关基因检测结果分析（附63例报告）

目的 探讨男性不育相关基因检测对不育原因的诊断价值，为助孕或使用供精提供理论依据。方法 收集男性不育患者血液样本并提取其中的DNA进行检测，经过片段化处理、末端修补、3’端加A、接头连接及扩增纯化等方法构建全基因组文库，并从中捕获、富集靶基因片段，构建靶基因文库。采用Illumina高通量测序平台对靶基因文库进行高通量测序。将测序数据与人类基因组GRCh37/hg19参考序列进行比对，筛选出相关变异。基因检测结果阳性代表本次检测检出已知的致病性或疑似致病变异，对判定为阳性的变异位点进行一代测序验证。结果 共检出6个基因9个变异位点为疑似致病突变或致病突变，因精液常规畸形率异常行基因检测的43例中有7例检出阳性基因（16.28%），其中因圆头精子症行检测的8例中检出3例（37.50%），非圆头精子症的35例中检出4例（11.43%），而因胚胎因素行基因检测的20例中有3例检出阳性基因（15.00%）。结论 在不育男性中行不育相关基因检测可发现遗传学因素导致不育的原因，可为进一步遗传咨询，包括继续常规助孕或使用供精提供理论依据。     

胃癌组织ras族基因突变与预后的关系

应用多聚酶链延伸反应－限制性片段长度多态笥分析法（PCR－RFLP）对88例福尔马林液固定，石蜡包埋胃癌组织C－Ha-ras第12位和13位，K－ras第12位和13位及N－ras第12位密码子的点突变进行检测，结果发现ras基因总突为18．2％（16．88），以c-Ha-ras第12位密码子突变率最高（13．6％），点突变的发生与肿瘤浆膜浸润，淋巴结转移，临床病理分期及术后生存期密切相关。以上结果提示，检测胃癌组织ras族基因点突变有助于判断胃癌患者的预后。     

老年冠心病患者载脂蛋白B100基因突变的研究

目的探讨老年冠心病发病基因水平上的原因,为冠心病的诊断提供理论依据。方法采用ＰＣＲ与限制性内切酶ＭｓｐⅠ酶切的方法对１８３例老年冠心病患者进行了载脂蛋白Ｂ１００基因突变的研究。结果７４例心绞痛病例中,基因水平有改变者为１８例（２４．４％）;１０例心肌梗死患者基因水平有改变者３例（３３．３％）,余者基因水平无改变。结论老年冠心病患者中心绞痛和心肌梗死两类冠心病部分病人病因与其基因水平碱基突变有关。     

老年人术后谵妄分析

对１２４０例老年手术病人进行了连续７ｄ的术后床边认识评分。结果发现有２００例病人发生术后谵妄,占总数的１６．３％,谵妄发生率为术后４ｄ内较高,以后逐渐下降。根据病人临床症状不同将２００例老年谵妄病人分为三类临床亚型,焦虑型：占发病人数０．３９％;安静型：占发病人数４．３％;混合型：占发病人数１８．０％;死亡人数占发病人数的４．０％。对老年术后谵妄的发病机理及防治进行初步探讨。     

冠心病患者血浆同型半胱氨酸及亚甲基四氢叶酸还原酶C677T基因多态性

目的比较不同冠心病类型及不同冠状动脉病变支数间血浆同型半胱氨酸水平差异,分析亚甲基四氢叶酸还原酶C677T基因突变对血浆同型半胱氨酸水平影响,及其与冠心病的关系.方法对经冠状动脉造影确诊的非冠心病74例、稳定型心绞痛32例、不稳定型心绞痛104例、急性心肌梗死25例采用改良高效液相色谱法测定同型半胱氨酸,聚合酶链反应-限制性内切酶法测定亚甲基四氢叶酸还原酶C677T基因型.结果血浆同型半胱氨酸水平冠心病组较非冠心病组显著增高,急性心肌梗死、不稳定型心绞痛组均较非冠心病组、稳定型心绞痛组显著增高,冠状动脉单支病变较正常、双支、三支病变组显著高.亚甲基四氢叶酸还原酶C677T基因纯合突变患者血浆同型半胱氨酸水平显著高于正常纯合子和杂合子患者.冠心病组亚甲基四氢叶酸还原酶C677T基因突变频率较对照组高,但无统计学意义.相关分析表明亚甲基四氢叶酸还原酶C677T基因突变与冠心病、同型半胱氨酸均无相关性.二元Logistic回归分析表明,高同型半胱氨酸血症致冠心病的OR值是1.138,亚甲基四氢叶酸还原酶C677T基因突变不是冠心病的独立危险因素.结论同型半胱氨酸可能是冠心病的独立危险因素,更可能是急性冠状动脉综合征的标志物.血浆同型半胱氨酸水平不与冠状动脉病变支数成正相关.亚甲基四氢叶酸还原酶C677T基因突变可能不是冠心病的独立危险因素及血浆同型半胱氨酸水平最关键的影响因素.     

Weight, the metabolic syndrome, and coronary heart disease in type 2 diabetes: associations among a national French sample of adults with diabetes-the ENTRED study

The authors examined whether obesity alone or as part of the metabolic syndrome (MS) increases coronary heart disease (CHD) risk in type 2 diabetes mellitus (T2DM) among 2970 adults aged 30-79 years in a French national sample. MS was defined as T2DM plus self-report of 2 or more of the following: body mass index >30 kg/m(2), diagnosed hypertension, or diagnosed dyslipidemia. A subsample with physician-reported data (n =841) was further classified with measured hypertension and dyslipidemia. Weight distribution included normal (21%), overweight (42%), and obese (37%). A 20% increased odds of CHD was estimated for every 5-kg/m(2) body mass index increase (P=.0001). MS was associated with a more than 2-fold higher risk of CHD compared with T2DM without MS (P<.0001, multivariate-adjusted [both samples]). With MS stratified by high-density lipoprotein cholesterol (<1.5 vs > or =1.5 mmol/L), compared with no MS, the odds ratio for CHD was 2.8 (normal-level high-density lipoprotein MS; 95% confidence interval, 1.8-4.5) and 1.5 (high-level high-density lipoprotein MS; 95% confidence interval, 0.8-2.9). The authors suggest that obesity alone--and particularly when the MS is present--increases CHD risk in patients with T2DM. High levels of high-density lipoprotein may modify this relationship.     

Lack of association between genetic variation in the beta3-adrenergic receptor gene and insulin resistance in patients with coronary heart disease.

The beta-adrenergic system plays a critical role in regulating lipolysis and thermogenesis. Recent studies have suggested that a missense Trp64Arg mutation in the beta3-adrenergic receptor gene is involved in visceral obesity and insulin resistance. We investigated the effect of this mutation on insulin resistance in patients with angiographically documented coronary heart disease ([CHD]n = 137) and normal subjects (n = 188). Plasma glucose and insulin responses to a 75-g oral glucose tolerance test and insulin resistance measured by the insulin suppression test, were determined in 58 (42%) patients with CHD and 121 (64%) controls. The genotype and allele frequency of the beta3-adrenergic receptor did not differ between patients with CHD and controls. The blood pressure, body mass index (BMI), waist to hip ratio, fasting plasma glucose, insulin, and lipid, and plasma glucose and insulin responses to the glucose load were relatively similar in subjects with and without the mutation in CHD and normal groups. The degree of insulin sensitivity, ie, the steady-state plasma glucose concentration, was not significantly different between subjects with and without the mutation in the CHD group (11.3 +/- 1.2, n = 11 v 11.9 +/- 0.6 mmol/L, n = 47, P = NS) and control group (8.4 +/- 0.7, n = 30 v 8.2 +/- 0.4 mmol/L, n = 91, P = NS). We conclude that Trp64Arg polymorphism of the beta3-adrenergic receptor gene does not likely play a major role in the development of CHD in the Chinese population. In addition, it appears to have no association with the insulin resistance syndrome in either CHD or non-CHD subjects.     

Relationship between metabolic syndrome (MS) and coronary heart disease (CHD) in an aged group

Our study aimed at investigating the relationship between metabolic syndrome (MS) and coronary heart disease (CHD) in aged patients, including 125 patients (age> or =60 years). Of them 78 cases belonged to the CHD group, which were subdivided into the CHD-MS group (38 cases) and the simple CHD group (40 cases); the other 47 patients without CHD belonged to the non-CHD group, which were subdivided again into the MS group (11 cases) and the control group (36 cases). Body mass index (BMI), blood lipids, blood uric acid, plasma fibrinogen, blood glucose and blood pressure of every patient were detected. The anatomy of coronary vessels was analyzed by selective coronary angiography to evaluate the relationship between MS and CHD. We found that the prevalence of MS in CHD group was significantly higher as compared to the groups not suffering from CHD (p<0.01). The CHD-MS group showed a higher prevalence of multivessel disease (p<0.05), unstable lesions (p<0.05) and needed more revascularization procedures (p<0.05) than the simple CHD group. The prevalence of CHD and the number of blocked coronary vessels were directly correlated with MS by Spearman correlation analysis (r=0.225, p<0.05; r=0.361, p<0.01). Logistic regression analysis demonstrated that both the risk of having future CHD and the number of blocked coronary vessels were directly correlated with MS (p<0.01; p<0.01), suggesting that MS can predict the prevalence and extent of future CHD in the elderly.     

伴有代谢综合征的冠心病患者高敏C反应蛋白浓度的变化

目的探讨伴有代谢综合征(MS)的冠心病患者血清中高敏C反应蛋白(hs-CRP)浓度的变化及其临床意义。方法选择179例冠心病患者为冠心病组,根据冠心病组患者是否伴有MS分为2个亚组,伴MS组(63例)和不伴MS组(116例)。同期选择健康体检者80例为正常对照组。测量身高、体重、血压,测定血糖、血脂和hs-CRP等相关指标,分析血清hs-CRP浓度与冠心病、MS的关系。结果冠心病组患者血清hs-CRP(1.77±0.96)mg/L较正常对照组(1.43±0.51)mg/L明显增高(P<0.05)。伴MS组血清hs-CRP(3.06±1.14)mg/L明显高于正常对照组,且差异有统计学意义(P<0.01);而不伴MS组血清hs-CRP(1.52±0.75)mg/L较正常对照组虽有增高,但差异无统计学意义(P>0.05)。血清hs-CRP浓度随着MS组分数目的增加而增高,且差异有统计学意义(P<0.05)。相关分析表明,血清hs-CRP与体重指数、收缩压、舒张压、TC、空腹血糖呈正相关(r=0.355,0.218,0.312,0.368,0.297,P<0.05),与HDL-C呈负相关(r=-0.221,P<0.05)。结论冠心病患者血清hs-CRP浓度增高,尤其以伴有MS的冠心病患者增高明显。     

黑龙江省东部地区汉族人群PCSK9基因第1外显子多态性与脂代谢的相关性

目的 研究黑龙江省东部地区汉族人群前蛋白转化酶枯草溶菌素(PCSK)9基因第1外显子多态性与脂代谢的相关性.方法 因心绞痛和(或)运动试验阳性及有冠脉管腔狭窄证据而需入院行冠状动脉造影者220例中,110例冠心病(CHD)者为病例组,110例非CHD者为对照组,检测PCSK9基因第1外显子基因多态性及血脂水平,并对基因多态性与血脂水平进行相关性分析.结果 共发现c.121C>G、c.299C>T、c.427-428insGCT、A53V、P56S和c.556A>G 6个突变位点,其中在A53V突变位点上发现CC和TC两种基因型,但未发现TT基因型.病例组和对照组各基因型间血清三酰甘油(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)水平差异具有统计学意义(P<0.05).病例组中TC基因型血清TG、TC和LDL-C水平均显著高于CC基因型(P<0.05);对照组中TC基因型血清TC水平显著高于CC基因型(P<0.05).结论 PCSK9基因第1外显子在黑龙江省东部地区汉族人群中存在多态性,且与CHD患者脂代谢有关.     

冠心病伴代谢综合征患者血清炎性因子与冠状动脉病变积分的相关性

目的:探讨伴代谢综合征(MS)的冠心病(CHD)患者血清白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)水平与冠状动脉(冠脉)病变积分(CAS)的关系。方法:共入选73例CHD患者,均行冠脉造影并计算CAS,同时根据患者是否伴发MS分为2组:CHD伴MS组(实验组)患者37例,单纯CHD组(对照组)患者36例,测定各组患者血清IL-6及TNF-α的水平并分析其与CAS的关系。结果:①实验组IL-6与TNF-α水平显著高于对照组(P<0.01);②实验组多重线性回归分析表明腰围、空腹血糖、IL-6及TNF-α与CAS呈显著正相关,偏回归系数分别为0.187(P=0.01)和0.119(P<0.01)。结论:CHD伴MS患者炎症反应更显著;IL-6和TNF-α与CAS呈正相关,与冠脉病变严重程度密切相关。     

Prevalence of coronary heart disease and carotid arterial thickening in patients with the metabolic syndrome (The ARIC Study)

We determined the prevalence of the metabolic syndrome (MS) with the criteria recommended by the National Cholesterol and Education Program, Adult Treatment Panel III report and estimated the magnitude of cross-sectional associations between the MS, coronary heart disease (CHD), and atherosclerosis in 14,502 black and white middle-age patients in the Atherosclerosis Risk in Communities Study. CHD was ascertained by standardized procedures and subclinical atherosclerosis was determined by measuring carotid intimal medial wall thickness using B-mode ultrasonography. The prevalence of MS was 30%, with substantial variation across race and gender subgroups. Among women but not among men, MS was significantly associated with increasing low-density lipoprotein cholesterol. CHD prevalence was 7.4% among those with the MS compared with 3.6% in comparison subjects (p <0.0001). After adjustment for established risk factors, subjects who had MS were 2 times more likely to have prevalent CHD than were those who did not have the syndrome. Among individuals free of CHD and stroke, after adjustment for age, gender, and race/center, the average intimal-medial wall thickness of carotid arteries was greater among those with versus those without MS (747 vs 704 mum, p <0.0001). Thus, MS was significantly associated with the presence of CHD and carotid intimal medial wall thickness. Identification of patients who have MS may provide opportunities to initiate CHD prevention strategies.