一家系人群遗传性神经源性腓骨肌萎缩症的临床研究

目的探讨婴儿期起病的腓骨肌萎缩症(Charcot-Marie-Tooth disease,CMT)一家系的遗传方式、临床特征、实验室及相关辅助检查、染色体核型及基因诊断等。方法按照遗传学标准家系分析纲要对患者家族成员作调查,分析临床特征,做详细的体格检查,部分行血常规、血生化、头颅CT、染色体核型检查。绘制家系发病图谱,并进行遗传学分析和基因诊断。结果先证者谱系代号为Ⅲ7,男性,5岁半。以发现双下肢进行性变细4年半就诊。家系4代41人中患病3人,死亡1例,均为男性,起病年龄为10月至1岁,所有患者均有肌肉萎缩,肢体无力,腱反射减弱或消失,跨阈步态。症状和体征随着年龄增长进行性加重。无明显智力低下。2名CMT患者及5名家系内表型正常者均发现17p11.2-p12区域内包含PMP22基因的重复突变,其亚型为CMT1A。结论 CMT是一种临床表型相似、由不同遗传方式引起的复杂性遗传性疾病,不同的临床表型由不同的基因突变引起;该家系遗传方式为常染色体显性遗传,临床分型为CMT1A;致病基因为17p11.2-p12区域内包含PMP22基因的重复突变。     

国人D17S30遗传多态性研究

采用基因扩增结合电泳银染技术分析１３０名汉族群体ＤＮＡＤ１７Ｓ３０遗传多态性，发现了１３个等位基因，基因频率为０．００７８～０．３５３８，有４２种基因型，其观察值和期望值经Ｘ２检验符合Ｈａｒｄｙ－Ｗｅｉｎｂｅｒｇ平衡定律；３个家系的９名成员Ｄ１７Ｓ３０遗传学分析，符合孟德尔遗传学规律。表明ＤＮＡＤ１７Ｓ３０在国人汉族群体具有高度多态性，在医学基因诊断、遗传学研究和法科学个人识别、亲权鉴定等领域是非常有价值的遗传标记。     

先天性长QT综合征家系的临床特点

的　通过对 3个先天性长QT综合征 (longQTsyndrome,LQTS)家系的调查 ,研究其发病情况、临床和心电图特点 ,推测其相应的基因型。方法　按常规采集 3个家系成员的临床病史 ,进行体格检查 ,采集静息心电图 ,测量QT间期和较正的QT间期。结果　 3个家系 4 3例中有 15例LQTS患者 ,11例可疑诊断。临床表现和心电图各异。结论　家系 1、家系 2和家系 3中LQTS患者的临床和心电图表现符合LQT2、LQT1和LQT3,可能为HERG、KVLQT1、和SCN5A的基因突变所致。     

基于基因组学探讨糖尿病肾病的发病原因及其与心血管疾病的关系

目的分析糖尿病肾病的发病原因及其与心血管疾病发生的关系,为二者的早期诊断和临床治疗提供依据。方法从Pubmed基因芯片公共数据库(GEO Database)中下载糖尿病肾病基因芯片数据,将数据导入R语言、QOE、STRING、GCBI及GenClip等分析软件或数据库,分析健康对照组与糖尿病肾病组的基因表达谱、基因功能、蛋白质相互作用网络、基因及信号共表达网络等,筛选两组之间的关键节点基因。结果糖尿病肾病患者肾小球组织的基因表达谱有明显改变;与健康人群相比,糖尿病肾病患者肾小球组织有844个差异表达基因;前20个差异表达基因蛋白-蛋白相互作用分析发现1个以TNNT2为核心的明显的蛋白相互作用网络;差异表达基因主要与炎症反应、应激反应等有关,且与MAPK信号通路关系密切。结论糖尿病肾病的发病原因主要与炎症反应、应激反应相关,且与心血管疾病的发生有一定的关系。     

甲状腺激素抵抗综合征一家系临床表型分析

目的对一甲状腺激素抵抗综合征患者及其家系成员进行调查分析。方法对患者及其14名家系成员详细询问病史,并行TT3、TT4、FT3、FT4、uTSH、TGAB及TMAB检查。结果家系中5名患者血清甲状腺激素水平升高,促甲状腺激素水平(TSH)也高于正常;他们的临床表现各异,有的表现为甲减,有的表现为甲亢,有的无任何临床症状。结论甲状腺激素抵抗综合征是一种临床表现复杂多变、极易漏诊、误诊的遗传性疾病。     

基于家系的腰椎间盘突出症患者强效基因筛选策略

目的对一个椎间盘退变疾病高发家系中4名椎间盘突出患者进行外显子基因测序,寻找此家系中可能导致椎间盘退变的强效致病基因,为研究椎间盘疾病的发病原因以及致病机制提供可能的帮助。方法收集西安交通大学第一附属医院、西安医学院第一附属医院、西安唐城医院自2010年1月至2016年12月之间就诊的腰椎间盘突出患者,对其进行详细的病史采集及家系调查,从中找到椎间盘退变疾病高发的1个家系。在此家系中,20～50岁成员中有4人为腰椎间盘突出症患者。利用外显子测序技术对这4例患者进行外显子基因分析,检测4例患者的共同外显子突变位点。结果最终在4个样本中检测出IGFBP6的共同突变Chr12:g.53494591T>C。Sanger法对其进行家系内验证,IGFBP6在4例患者中皆表现为C/T型,而在5例正常对照者中皆表现为T型。最后在正常对照组200例中进行验证,未发现此位点变异。结论 IGFBP6的突变位点c.T430C(p.S144P)(Chr12:g.53494591T>C)可能是导致该家系中椎间盘突出疾病高发的因素;外显子测序技术可以应用于复杂疾病致病基因的发掘。     

一脆性X综合征家系的基因诊断及其动态突变

本文报告了使用探针pX6对一脆性X家系进行了分子杂交分析研究。结果表明:女性携带者的（CGG）的较小扩增遗传到子代出现（CGG）重复序列大量扩增,伴随CpG岛甲基化,从而导致FMR-1基因失活。     

Becker型肌营养不良一家系分子遗传学研究

目的 应用分子遗传学方法对患者在基因水平上作出诊断。方法 应用ｄｙｓｔｒｏｐｉｎ ｃＤＮＡ１４ｋｂ探针（包括６个亚探针１－２ａ，２ｂ－３，４－５ａ，５ｂ－７、８、９－１４）与一名１８岁的临床表现温和肌营养不良患者及２例对照者（１例正常２５岁男性及１例１２岁ＤＭＤ患者）的基因组ＤＮＡ／Ｈｉｎｄ Ⅲ片段进行Ｓｏｕｔｈｅｒｎ印迹分析。结果 前者在与亚探针５ｂ－７杂交中，发现其１．５ｋｂ，０．５ｋｂｖｓｕ     

人类OPRM1-EXON1的克隆与探针的制备

目的　克隆、测序人类 OPRM1- EXON1,并用非同位素 生物素标记法对该基因进行标记、制备探针,用于OPRM1- EXON1的表达及功能研究。方法　通过PCR法扩增目的基因片段,并将其连接到pGEM- T载体上,转入感受态细胞中进行重组并克隆,经酶切和基因测序进行鉴定,用非同位素 生物素标记法进行探针的标记与制备。结果　经过PCR扩增的目的基因片段大小(2.2 kb)与理论上片段大小一致,经过测序证实其序列与 NCBI提供的序列相同,用此片段成功的制备了用于研究阿片受体基因的探针。结论　从基因组中成功克隆了人类 OPRM1- EX ON1,并制备了探针,为深入研究吗啡依赖相关基因及基因表达创造了必要的条件。     

一个遗传性共济失调家系报告

本研究小组获得常染色体显性遗传共济失调家系 ,为六代 17例患者 ,存活的患者 6人 ,其中男性 3人 ,女性 3人 ,见图 1。先证者 ,男 ,4 0岁 ,自 2 5岁始出现行走不稳。原为木匠 ,早年因定点不稳和辨距不清而四处求医。症状逐渐加重 ,已发展至依赖木棍支撑行走。四肢震颤 ,肌张力高。近两年来出现言语含混不清 ,双眼可见持续的水平性眼球震颤。共济及平衡功能差。患者智力稍有退化。肢体感觉和四肢腱反射未见异常。家系中存活的患者 ,病情严重程度不一 ,但均表现为共济和平衡功能差。其中Ⅳ1 3伴有渐进的四肢发硬、关节疼痛及完全行走困难。家…     

THRB基因第7、第10外显子遗传标记与抑郁症的关联和连锁不平衡研究

目的从单核苷酸多态性(SNP)和单倍型分析的角度入手分析抑郁症患者甲状腺激素β受体(THRB)基因与抑郁症之间的关系。方法入组抑郁症患者及健康对照者各50例,均为中国陕西籍汉族人。提取基因组DNA,对THRB基因第7、第10外显子测序,并对患者进行汉密尔顿抑郁量表(HAMD)、汉密尔顿焦虑量表(HAMA)评分。测序结果进行序列比对,采用SPSS软件进行统计分析,采用SHEsis在线分析系统、LDA 1.0软件、Haploview 4.0软件进行单倍型和连锁不平衡分析。结果全部样本THRB基因第7外显子上未发现SNP,抑郁症患者组第10外显子上发现G1457T和G1671A两处SNP,健康对照组第10外显子上发现G1671A一处SNP,组间分布无统计学差异(P>0.05)。THRB基因G1671A与G1457T两处SNP构成的单倍型位于存在强烈连锁不平衡关系的单倍型域内,抑郁症患者组中存在3种单倍型分布,表现为连锁不平衡关系;健康对照组中发现两种单倍型分布,不存在连锁不平衡关系。携带THRB基因G1671A杂合子的抑郁症患者HAMD量表总分高于野生型纯合子个体;携带THRB基因G1457T杂合子的抑郁症患者HAMD量表抑郁情绪、HAMA量表抑郁心境得分高于野生型纯合子个体,HAMD迟缓因子得分低于野生型纯合子个体。结论中国陕西省汉族抑郁症患者THRB基因第7、第10外显子上存在不同于现有报道的SNP分布,G1671A与G1457T两处SNP位点及其构成的单倍型与抑郁症不存在关联和连锁不平衡,携带THRB基因G1671A杂合子和/或携带THRB基因G1457T杂合子的抑郁症患者有不同于携带上述两位点野生型纯合子个体的临床表现型。     

MITOCHONDRIAL DNA POLYMORPHISM IN CONTROL REGION FROM CHINESE YUGU POPULATION

Objective To investigate the mitochondrial DNA sequence polymorphism sites in Chinese YUGU ethnic group and to provide basic data used in forensic purpose. Methods Genomic DNA was extracted from the hole blood of 100 unrelated individuals of Chinese YUGU ethnic group by standard chelex-100 method. The sequence polymorphism sites was determined by PCR amplification and direct sequencing. Results 54 polymorphic sites were noted in mtDNA np16091-16418 region, and 46 haplotypes were identified. The genetic diversity was calculated to be 0. 9691, and the genetic identity was calculated to be 0. 0406. Conclusion There are some particular polymorphism sites in Chinese YUGU ethnic group. The results suggest that sequence polymorphism from np]6091 -- 16418 in human mitochondrial DNA can be used as a biological marker for forensic identity.     

陕西地区汉族群体Y染色体STR基因座多态性研究

目的　研究 14 0名陕西地区汉族男性个体Y染色体 6个STR基因座的遗传多态性 ,为法医学实践和群体学研究提供基础数据。方法　应用Y PLEXTM6荧光标记复合扩增系统 ,PCR扩增后用ABI 310 0Avant遗传分析仪进行基因扫描和基因分型。结果　 6个基因座分别检测出 4、6、7、8、4、37种等位基因 ,基因多样性 (GD)从 0 .35 94～0 .95 6 5。在 14 0名个体中共检测出 135种不同的单倍型 ,其中 131个只出现 1次 ,单倍型多样性 (HD)为 0 .992 3,个体识别能力 (DP)为 0 .96 4 3。结论　这 6个Y染色体STR基因座具有较好的多态性 ,联合单倍型具有较高的个体识别能力和非父排除率 ,适用于法医学实践和群体遗传学研究     

DIG－DNA探针检测宫颈癌组织中HPV－16的E_7转化基因

应用地高辛配基（Ｄｉｇ－１１－ｄＵＴＰ）标记人乳头瘤病毒１６型（ＨＰＶ－１６）Ｅ７转化基因作探针，分别检测了宫颈癌活检组织、宫颈炎及正常宫颈脱落细胞ＤＮＡ。其检出率为：宫颈癌中４２．１％（２４／５７）、宫颈炎中２．５％（１／３９）、正常宫颈中５．５％（１／１８）。宫颈组织中ＨＰＶ－１６Ｅ７转化基因的检出率较高，提示该转化基因与宫颈癌的发生可能密切相关。本研究也证实ＤＩＧ配基标记的Ｅ７基因探针具有特异、敏感、安全、稳定和能反复使用的优点，适合基层使用。     

ASSOCIATION ANALYSIS OF POLYMORPHISMS IN SIX GENES WITHIN THE GH/IGF-1 AXIS IN PATIENTS WITH IDIOPATHIC SHORT STATURE IN THE CHINESE HAN POPULATION

Objective To investigate relationships of polymorphisms in six genes ( GHR,IGF-1,IGF-1R,IGFBP-3,JAK2,and STAT5b) in the growth hormone ( GH)/insulin-like growth factor-1 (IGF-1) axis with idiopathic short stature (ISS) in the Chinese Han population. Methods A casecontrol study was carried out on a cohort of 198 ISS patients and 306 healthy controls.A total of 106 tagging single nucleotide polymorphisms (tagSNPs) from the six genes were selected from the HapMap ( haplotype map of the human genome ) Han Chinese in the Beijing subset.Results of genotyping conducted by highthroughput Illumina GoldenGateTM Assay were analyzed by statistical software. Results Both individual tagSNPs and haplotypes showed an association with ISS in the Hun Chinese population ( P ＜ 0.05 ).For each single test,both allele and genotype were tested.By allele frequency analysis,six positive SNP sites ( rsNo.1,rsNo.2,rsNo.3,rsNo.4,rsNo.5,and rsNo.6 ) of 3 genes ( JAK2,IGF-1R,and GHR) were found having associations with ISS. By genotype frequency analysis, there were significant differences between the patient and control groups in the following SNP sites:4 sites in JAK2 gene ( rsNo.1,rsNo.2,rsNo.3,and rsNo.4 ) and 1 site in GHR gene ( rsNo.6 ).The risk which affected ISS was found related to the JAK2 gene in 4 sites ( increase in rsNo.1 and decrease in rsNo.2,rsNo.3,and rsNo.4) and to the GHR gene in 1 site (decrease in rsNo.6).They were four haplotypes in gene of IGF-1R as “ TGC","CGCT",”TA",and "CA",one haplotype in IGFBP-3 as "TA",and one haplotype in JAK2 as "CTG",which revealed high significance for risks of affecting ISS. At last,multivariate logistic regression analysis of specific site rsNo.6 of the GHR gene revealed that the serum IGF-1 was related to genotypes AA and AC,with genotype CC as the reference ( P =0.015). Conclusion Genetic variances in six genes within the GH/IGF-1 axis may be important etiological factors for ISS in the Chinese Han population.     

GENETIC POLYMORPHISM OF SIX Y CHROMOSOMAL STR IN CHINESE HUI ETHNIC GROUP

Objective To study genetic polymorphism of 6 Y chromosomal STR in Hui ethnic group living in Ningxia Hui ethnic autonomous region, in order to evaluate their usefulness in forensic science and enrich the Chinese genetic information resources. Methods We investigated 101 unrelated, healthy, male individuals of Hui ethnic group and studied their allelic frequency distribution and haplotype diversity of 6 Y chromosomal STR. Primer for each loci was labeled with the fluorescent by FAM (blue) or TAMRA(yellow). The data of Hui ethnic group were generated coamplification, GeneScan, genotype, and genetic distribution analysis. Results 31 alleles and 43 phenotype(DYS385) were detected, with the frequencies ranging from 0. 0099- 0. 7129. Out of a total of 101 individuals, 96 showed different haplotypes; 91 were unique; 5 were found 2 times. The haplotype diversity for 6 Y-STR loci was 0. 9990. Conclusion The date obtained can be valuable for individual identification, paternity testing in forensic fields and for population genetics because of 6 Y-STR loci high polymorphism.     

腺病毒携带GFP在豚鼠耳蜗基因转导的形态结构与功能改变

目的　带有绿色荧光蛋白 (GFP)基因的腺病毒注入豚鼠耳蜗后 ,观察不同时间段GFP基因的表达和分布情况及手术操作对听力的影响 ,为内耳基因治疗提供理论依据。方法　 1 5只杂色豚鼠术前及术后行听性脑干反应 (ABR)和畸变耳声发射 (DPOAE)检查 ,空白对照组经圆窗或底回钻孔注入人工外淋巴液。实验组注入带有GFP基因的腺病毒。分别于 3、5、7和 1 0d后取材 ,耳蜗标本经硝酸银染色后铺片。结果　腺病毒注入耳蜗后对听阈影响不大。术前术后的DP值有差异。荧光显微镜下可见 3d组表达产物最高。 7d后逐渐减弱 ,1 0d后更弱 ,表达产物主要分布于支持细胞、螺旋神经节细胞和基底膜下的间皮细胞。对照组均无绿色荧光。结论　通过圆窗或底回注入腺病毒对听阈没有影响。单一位点的接种就能使神经营养因子通过耳蜗液扩散到整个耳蜗。有效的基因转移在耳蜗是可行的 ,但表达相对短暂。     

DISTRACTION OF OSTEOGENESIS AND LENGTHENING OF CRANIOFACIAL BONE-PRELIMINARY REPORT OF AN EXPERIMENTAL RESEARCH

Themethodforgradualcallusdistractionofosteogenesis(Do)inenchondralbonehasbeenu-niversallyappliedontheorthopedicclinics.DowasfirstlyusedonfemurlengtheningbyanItal-ian,Codivilla(19O5),onthepatientsofunequiva-lentofstature,whilethetibiawaslengthenedbyAbbotin1927[1i.Becauseoftheseverecomplica-tionsasedema,necrosisofskin,infections,angu-lardeformity,andthedelayedmineralizationofthedistractedbone,themethoddidn'tarousedthein-terestofpeopleuntilearly2Othcentury,195Ost2i.ItwasIlizarov(aRussiansurge…     

ROLE OF ANTISENSE AND DECOY OLIGONUCLEOTIDE OF NF-κB IN VESSEL STENOSIS AND NEOINTIMA FORMATION IN BALLOON-INJURED RAT ARTERY

Objective To examine antisense and decoy oligonucleotides of nuclear factor kappa B in vivo effects on intima proliferation and balloon-injured monocytes chemotactic protein-1 ( MCP-1 ) and extracellular signal regulated kinase-2 ( ERK2 ) expression in the carotid artery of rats. Methods Sprague-Dawley rats underwent balloon-dilation injury of the left carotid artery. Rats are divided into 7 groups ( n = 18 ) and each group includes 6 time points (6 h, 1, 3, 5, 7, 14 d) ( n =3). Uninjured right carotid artery of the same rat was used as controls. Results In model group, sense group and scramble group, vessel intima area , media area and intima/media ratio increased after 5 d and reached the maximum after 7 d. The effect of antisense plus decoy group on intimal hyperplasia was more obvious than that of antisense group and decoy group alone. MCP-1 mRNA expression was increased expression continuously at 3, 5 and 7 d and decreased at 14 d. Compared with model group, sense group and scramble group, antisense group, decoy group and antisense plus decoy group had lowered MCP-1 mRNA expression in each time point ( P ＜ 0. 05 ). NF-κB p65 was dispersed positive stain 6 h after injury and increased after 1 d and peaked at 7 d, but the protein expression was weak at 14 d. ERK2 protein synthesis increased at 1 d and reached the peak at 7 d, while protein expression after 14 d was similar to that at 7 d. Treatment of antisense group,decoy group and antisense plus decoy group inhibited protein synthesis more significantly than those of model group,sense group and scramble group ( P ＜ 0. 05 ). Conclusion NF-κB modulates genes expression and protein synthesis of MCP-1 and ERK2. Celluar proliferation in vessel wall was dynamically changed after balloon angioplasty injury. Antisense and decoy oligonucleotide of NF-κB by local lipofectamine transfer inhibit NF-κB activating gene modulation and neointimal hyperplasia.     

IDENTIFICATION OF HUMAN MURR1, THE HOMOLOGUE OF MOUSE IMPRINTED Murr1 GENE

Different fromthe biallelic expression mannerof most common autosomal genes,i mprinting is aspecial genomic expression phenomenon,of whichthe i mprinted genes show parental-specific expres-sion only according to their parental origin[1].Cor-rect i mprintingis i mportant inthe nor mal embryon-ic development,postnatal growth,as well as behav-ior of ani mals.Dysregulation of i mprinting hasbeen i mplicated in the pathogenesis of congenitaldiseases and cancer[2-3].To date,78kinds of mousei mprinte…