Association between SNP rs10569304 on the Second Expressed Region of Hole Gene and the Congenital Heart Disease

The correlation of single nucleotide polymorphism (SNP) rs10569304 on the second expressed region of hole gene and congenital heart disease (CHD) of human being, and the effect of hole gene on CHD were investigated. 179 patients with CHD as CHD group and 183 healthy people as control group were selected in the case-control study. DNA was abstracted from the peripheral blood by phenol-chloroform method. Primer was designed for the flanking sequence of SNP rs10569304 on the second expressed region of hole gene. The genotype was identified by PCR degenerative acrylamide electrophoresis with amplification products. Then the three amplification products received sequencing. By chi-square test, the genotype frequency and allele frequency in CHD group and control group were analyzed. There was insertion-deletion (GCC/-) of SNP rs10569304 which corresponded to alleles of A and B in Southern Chinese people. The genotype frequency and allele frequency in control group and CHD group were met the Hardy-Weinberg equilibrium. By chi-square test, in control group and CHD group, the genotype frequency of AA (insertion homozygous), AB (insertion-deletion heterozygous) and BB (deletion homozygous) was 21.31%, 54.09%, 24.59% and 16.75%, 46.36%, 36.87%, respectively. The distributional difference of genotype frequency had statistical significance (χ2=6.51, P<0.05); The allele frequency of A and B was 48.36% and 51.64% in control group, 39.94% and 60.06% in CHD group, respectively. The distribu- tional difference of allele frequency had statistical significance (χ2=5.20, P<0.05). Meanwhile, by contrast with the control group, the BB genotype frequency and B allele frequency in CHD group was higher, but the AA and AB frequency was lower. There was higher risk to suffer from CHD involving B allele. BB genotype had 1.907-fold increased risk of developing CHD according to AA genotype (P<0.05). It is concluded that there is insertion-deletion (GCC/-) of SNP rs10569304 in the Southern Chinese people, and the people whose hole gene involving BB genotype have higher risk to suffering from CHD.

HOLE基因外显子2区SNP与先天性心脏病的关联研究

目的:研究HOLE基因外显子2区SNP与人类先天性心脏病(CHD)的相关性。方法:以病例组179例CHD患者和对照组183例门诊体检健康人为研究对象,酚-氯仿法提取外周血DNA,PCR扩增产物经变性丙烯酰胺电泳鉴定基因型,应用卡方检验分析组间基因型频率及等位基因频率差异。结果:(1)粤黔两地汉族人群中存在HOLE基因SNPrs10569304的插入缺失(GCC/-)多态性,对应为等位基因A/B。(2)对照组和病例组的基因型频率及等位基因频率符合Hardy-Weinberg平衡,两组AA、AB和BB三种基因型频率分别为21.31%、54.09%、24.59%和16.75%、46.36%、36.87%,经卡方检验基因型分布差异有统计学意义(χ2=6.51,P<0.05),与对照组相比,病例组中BB基因型频率升高,AA、AB基因型频率降低。(3)对照组与病例组A和B等位基因频率分别为48.36%,51.64%和39.94%,60.06%,病例组B等位基因频率显著高于对照组,分布差异有显著性(χ2=5.20,P<0.05)。结论:粤黔两地汉族人群中HOLE基因SNPrs10569304存在插入缺失(GCC/-)多态性,具有BB基因型的个体罹患CHD的危险度增高。

人hole基因的克隆和序列分析

目的克隆人的hole基因,并进行生物信息学分析。方法以胚胎心脏cDNA文库为模板,进行PCR扩增得到hole基因的ORF。并进行亚克隆入PMD18-T载体,转化大肠杆菌DH5α,用Xho1进行酶切和测序鉴定阳性克隆。利用Internet和GeneBank数据库对测序结果正确的hole基因进行生物信息学分析。结果克隆了hole基因,测序结果正确;生物信息学分析表明,hole基因开放读码为960 bp,编码319个氨基酸;同源性分析表明,人的hole与鼠、鸡hole蛋白具有高度同源性。多种组织的半定量RT-PCR研究表明,该基因在心脏、肝脏、肺、脑、肾脏、脾脏、胰脏均有表达,其心脏表达最高。结论成功克隆hole基因,对其功能进行了初步预测,为进一步的功能研究打下基础。

Determining the Scour Dimensions Around Submerged Vanes in a 180°Bend with the Gene Expression Programming Technique

Submerged vanes are installed on rivers and channel beds to protect the outer bank bends from scouring.Also,local scouring occurs around the submerged vanes over time,and identifying the effective factors on the scouring phenomena around these submerged vanes is one of the important issues in river engineering.The most important aimof this study is investigation of scour pattern around submerged vanes located in 180°bend experimentally and numerically.Firstly,the effects of various parameters such as the Froude number(Fr),angle of submerged vanes to the flow(α),angle of submerged vane location in the bend(θ),distance between submerged vanes(d),height(H),and length(L)of the vanes on the dimensionless volume of the scour hole were experimentally studied.The submerged vanes were installed on a 180°bend whose central radius and channel width were 2.8 and 0.6 m,respectively.By reducing the Froude number,the scour hole volume decreased.For all Froude numbers,the biggest scour hole formed atθ=15°.In all models,by increasing the Froude number,the scour hole volume significantly increases.In addition,by increasing the submerged vanes’length and height,the scour hole dimensions also grow.Secondly,using gene expression programming(GEP),a relationship for determining the scour hole volume around the submerged vanes was provided.For this model,the determination coefficients(R2)for the training and test modes were computed as 0.91 and 0.9,respectively.In addition,this study performed partial derivative sensitivity analysis(PDSA).According to the results,the PDSA was calculated as positive for all input variables.

按揉“委中”穴影响腰椎间盘突出症患者血浆内皮素和降钙素基因相关肽含量及临床疗效研究

目的基于"腰背委中求"对按揉"委中穴"治疗腰椎间盘突出症患者血浆内皮素(ET-1)和降钙素基因相关肽(CGRP)含量变化,探讨按揉委中穴治疗腰腿痛的作用机制及临床疗效。方法采用按揉委中穴推拿治疗30例腰椎间盘突出症腰痛患者,检测治疗前后的ET-1与CGRP的数值,并与对照组进行比较分析。结果检测结果发现腰痛患者ET-1高于对照组,而CGRP低于对照组。治疗前后比较,两者均有显著差异(P<0.01)。结论研究提示按揉委中穴对腰椎间盘突出症腰腿痛治疗有效,且对ET-1与CGRP平衡有调节作用。

心肌特异性Hole转基因小鼠模型的构建与鉴定

目的:建立在小鼠心肌细胞中特异性过表达Hole转基因小鼠。方法:构建心肌特异性过表达Hole的转基因载体,显微注射导入小鼠受精卵,通过胚胎移植,获得转基因首建者小鼠。利用PCR检测Hole基因整合情况,并通过实时定量PCR检测Hole基因过表达效率。结果:PCR检测发现有6只小鼠在其基因组上整合有Hole载体基因。实时定量PCR结果显示,在心脏组织中Hole基因的转录本表达明显升高。结论:成功建立了在小鼠心肌细胞中特异性过表达Hole转基因小鼠,为研究Hole基因在心脏发育与相关疾病中的功能及机制提供了动物模型。