11个Fabry病家系的α-半乳糖苷酶A活性及GLA基因检测

目的 建立Fabry病α-半乳糖苷酶A（α-gal A）酶活性及基因检测体系,并对基因型临床表型进行分析。方法 检测11个Fabry病家系先证者及家系成员外周血粒细胞α-gal A活性及GLA基因。酶活性检测采用底物荧光法,基因检测采用PCR直接测序法,并进行临床评估。结果 在11个Fabry病家系中检出9种GAL基因突变,包括5个错义突变（R301Q、I91T、G132R、F273L、D165Y）,2个无义突变（W236X、R342X）,1个单碱基缺失（1082delG）和1个大片段缺失（44 bp nt．1077）,其中4种为新突变（D165Y、F273L、1082delG、44 bp nt．1077）。11个家系中通过基因及酶活性检测,确诊男性半合子13例,女性杂合子12例。男性半合子α-gal A酶活性显著下降,女性杂合子α-gal A酶活性部分下降,1/4女性杂合子的α-gal A酶活性处于正常范围内。结论 确诊了11个Fabry病家系的GLA基因突变类型,并筛出所有家系中先证者以外的患者14例。外周血粒细胞α-gal A酶活性和GAL基因检测是筛查和诊断Fabry病的有效手段。     

一个Fabry病家系的基因突变和临床表型研究

目的对一个临床表现以肾脏损害为主的Fabry病家系进行调查,分析其基因突变和临床特点。探讨不同突变类型对临床表型的影响。方法收集该例先证者及其家族成员的临床资料。采集先证者及其姐姐和一名健康对照者的外周血,应用荧光底物法检测a半乳糖苷酶A（toga[A）的酶活性;提取血液基因组DNA,PCR分段扩增α—galA基因的7个外显子,产物纯化后进行DNA测序,检测是否存在突变位点。结果①临床调查患者家系发现5名男性患者,均发展至终末期肾脏疾病（endstage renaldisease,ESRD）,伴有肢端疼痛、皮肤血管胶质瘤及少汗等Fabry病典型表现;3名女性杂合子,临床表现各异;②regalA活性检测示先证者酶活性明显下降,其姐姐α-galA活性在正常范围;③基因检测发现先证者regalA基因第2外显子的第112个密码子上的碱基胞嘧啶（c）被胸腺嘧啶（T）替代,因此导致由原本翻译的精氨酸变成了半胱氨酸（p．R112C）,形成错义突变,从而导致α-galA酶活性的降低或缺乏。结论发现了一个新的Fabry病家系,并明确了该家系的基因突变特点。在此家族中,我们发现除了外胚层组织损害,α—galA基因错义突变p．R112C亦可导致中胚层组织（如肾脏）的严重损害。     

成人经典型Bartter综合征家系CLCNKB基因突变的研究

目的 分析1个成人经典型Bartter综合征家系CLCNKB基因突变特点。 方法 用PCR方法对先证者CLCNKB基因19个外显子及侧翼序列进行扩增,PCR产物纯化后直接测序或构建T-A克隆测序检测其基因变异。 结果 先证者CLCNKB基因表现为G433E和cDNA 753delG复合杂合突变。家系分析表明,杂合错义突变G433E来自父亲,杂合缺失突变cDNA 753delG来自母亲。患者弟弟携G433E,其妹妹正常。正常对照100条染色体中未发现同样变异。 结论 在1个成人迟发经典型Bartter综合征家系中发现了CLCNKB基因2个突变位点,经检索文献及人类基因突变库（HGMD）,cDNA 753delG为新发现突变位点。     

中国人恶性高热家系蓝尼定受体-1基因的筛查

目的 筛查中国人恶性高热(MH)家系的蓝尼定受体-1(RYR1)基因.方法 提取确诊为MH的先证者及家系其他成员外周血白细胞的基因组DNA,采用PCR扩增先证者RYR1基因的部分外显子,并进行直接测序,采用Fok Ⅰ限制酶分析验证先证者及家系其他成员基因突变情况.结果 先证者RYR1基因第6 724位碱基C突变为T(c.6724 C＞T),所编码第2 206位氨基酸由苏氨酸变为甲硫氨酸(p.T 2206 M),此突变在白种人已报道;先证者的4个子女中有2个为该错义突变携带者,为MH易感者.结论 中国人MH易感者携带与白种人MH易感者相同的RYR1基因突变.     

两个遗传性并多指(趾)家系致病基因突变分析

目的探讨2个并多指(趾)畸形(SPD)家系的致病基因。方法收集2019年1月、2020年12月就诊于临沂市人民医院的2个SPD家系的临床资料, 采集先证者及家系成员静脉血样本, 提取基因组DNA, 对先证者行全外显子组测序筛选候选基因变异;采用Sanger测序对2个家系成员验证其突变位点;采用生物信息学软件PolyPhen-2和PROVEAN对突变位点的致病性进行预测分析, 结合美国医学遗传学与基因组学学会(ACMG)指南对突变位点进行致病性判断。结果家系1三代成员中共有5例患者(男2例、女3例), 先证者为8岁女性, 表现为右手第3、4指并指, 指蹼融合和远端指甲融合, 其余手指活动自如, 双脚未见异常;家系2三代成员中共有4例患者(均为女性), 先证者为4岁女性, 表现为双手第3、4指并指, 示指侧弯。全外显子组测序分别在2个SPD家系中检出同源盒D13(HOXD13)基因c.917G>A和c.917G>T突变, 且2个突变均呈现基因型-表型共分离, 其中HOXD13基因c.917G>T突变未见数据库收录, 为新发杂合错义突变。生物信息学软件预测这2个突变位点均为...     

VHL基因N131Y突变导致家族性胰腺多发囊肿

目的探讨1个单纯表现胰腺多发囊肿的Von Hippel-Lindau(VHL)综合征家系的VHL基因变异情况。方法调查1个VHL病家系临床资料,绘制树状图;抽取该家系3位成员外周血,提取基因组DNA,应用聚合酶链式反应(PCR)以及实时定量PCR对VHL基因进行点突变以及基因缺失进行筛查。结果该家系中仅先证者和其母亲表现为胰腺多发囊肿,且基因检测结果显示先证者和其母亲均存在VHL基因编码区第391位核苷酸A突变为T,导致第131号编码氨基酸由天冬酰胺变成了酪氨酸,为VHL基因未曾报道过的突变位点;而其父亲均无异常表现。结论 VHL综合征中,VHL基因2号外显子发生该位点突变,则仅表现为胰腺多发囊肿;对于单纯表现为胰腺多发囊肿的患者,尤其是青年或无胰腺疾病家族史者需考虑VHL综合征的可能。     

经典型Bartter综合征家系CLCNKB基因突变分析

目的 研究一个经典型Bartter综合征家系CLCNKB基因突变情况。 方法 提取该家系各成员患者外周血淋巴细胞基因组DNA,应用PCR扩增CLCNKB基因全部外显子及侧翼序列,并直接测序检测突变。选取50例无亲缘关系的健康人作为对照。 结果 在患者中检测到1个杂合（错义）突变,其第4号外显子,第482位碱基T→G突变,造成第161位氨基酸由亮氨酸变为精氨酸（482T>G,L161R）;家系中母亲为杂合突变（L161R杂合突变）,父亲未发现突变;查阅国内外文献及人类基因突变数据库,L161R未见报道,属新发现的突变。 结论 发现了一种新的CLCNKB基因突变：L161R。     

家族性青少年高尿酸血症肾病家系患者尿调节素编码基因突变位点研究

目的本文报道家族性青少年高尿酸血症肾病(FJHN)一家系的尿调节素(UMOD)编码基因新的位点突变并结合文献复习,以期引起肾脏病学者对该病的足够重视。 方法收集、核实和整理分析1例FJHN先证者的临床特征、实验室检查及该家系其他成员相关的临床资料;检测先证者及其长子和外甥的UMOD编码基因外显子2～5变异情况。 结果先证者呈现典型的FJHN临床表现,包括青年发病,显著性高尿酸血症、痛风性关节炎和痛风石,早期即有夜尿增多等尿浓缩功能减退的表现,肾功能损害持续缓慢进展,至40岁左右时发展为终末期肾病。家系调查及基因突变检测显示,家系三代中至少有高尿酸血症12人,其中3人已死于尿毒症,先证者也已进入尿毒症期,并正在接受血液透析治疗。先证者及其长子经基因检测均有相同的基因突变：UMOD编码基因位第4外显子上第854碱基出现C/A嵌合子(正常参考基因碱基为C)变异,使氨基酸序列第285位丙氨酸(A,GCG)变异为谷氨酸(E,GAG)。无高尿酸血症的先证者外甥经基因检测未发现基因外显子2～5变异。 结论家族性青少年高尿酸血症肾病可能与UMOD编码基因位第4外显子上第854碱基变异(此为一新发现的基因突变)有关,对有显著高尿酸血症、痛风,尤其是有慢性肾脏病家族史的青(少)壮年患者,应考虑有无FJHN的可能性,通过医学影像学、肾活检和(或)UMOD基因的检测,尽早明确诊断、避免误诊、漏诊。     

中国人遗传性非息肉病性结直肠癌家系错配修复基因突变的研究

目的探讨中国人遗传性非息肉病性结直肠癌(HNPCC)家系中hMLH1、hMSH2基因遗传性突变情况。方法取14个符合中国人HNPCC标准的HNPCC家系肿瘤先证者外周血DNA样本,聚合酶链反应扩增hMLH1、hMSH2基因共35个外显子,应用变性高效液相色谱技术(DHPLc)结合DNA测序法检测突变。结果14个家系中共发生41个大肠癌和19个肠外恶性肿瘤,其中胃癌是最常见的肠外恶性肿瘤类型。14个患者中检测到分属于9个家系的12个遗传性单个碱基改变,其中8％为无义突变,25％为错义突变,其余42％为单核苷酸多态,17％为内含子区的单碱基改变,8％为同义突变。结论(1)应用DHPLc成功检测到hMLH1、hMSH2基因杂合性突变。(2)符合中国人HNPCC标准家系约有1／3可检出hMLH1、hMSH2基因遗传杂合性突变,其中错义突变较多见。     

中国人遗传性非息肉病性结直肠癌临床及遗传学特征分析

目的分析中国人遗传性非息肉病性结直肠癌(hereditary nonpolyposis colorectal cancer,HNPCC)家系的临床及遗传性突变特点。方法收集整理符合中国人HNPCC诊断标准的31个家系资料,应用PCR及变性高效液相色谱分析(denaturing high-performance liquid chromatography,DHPLC)筛查hMLH1和hMSH2基因的突变,对DHPLC图形异常的样本进行测序。结果31个家系中共发生136例次恶性肿瘤(多原发肿瘤14例),其中结直肠癌106例次,占所有肿瘤患者的77．9%,诊断年龄均数为48．6±29．0岁；胃癌14例。肿瘤先证者中共检出分属于17个不同家系的23个碱基变异位点,经DNA序列分析,证实10个家系存在10个不同的碱基突变(10/31,32．9%),其中3个为同义突变未引起蛋白质序列的改变,另外7个为病理性改变,分别为错义突变、无义突变、移码突变。这10个突变中7个位点为首次报道。外显子区检测到5个已知SNP,内含子区检测到8个碱基改变。结论(1)符合中国人HNPCC标准家系约有1/3可检出hMLH1、hMSH2基因遗传性种系突变。(2)中国人HNPCC家系以左半结肠癌和直肠癌多见。     

A novel GLA mutation in a Fabry family with glucose-6-phosphate dehydrogenase deficiency

Fabry disease is an X-linked lysosomal disease caused by mutations of the alpha-galactosidase A (GLA) gene at chromosome subband Xq22.1. To date, more than 600 genetic mutations have been identified to determine the nature and frequency of the molecular lesions causing the classical and milder variant phenotypes and for precise carrier detection. We report here a Fabry family (mother, son and daughter) where the alpha-galactosidase A defect was associated with a glucose-6-phosphate dehydrogenase (G6PD) deficiency. Mutation analysis revealed for the GLA gene the presence of a new mutation, i.e., a small deletion (c.452delA) on exon 3 and for the G6PD gene the presence of 2 mutations, p.V68M (G6PD Asahi, G6PD A+) and p.N126D (G6PD A+) on exon 3 and exon 4, respectively.     

14例Fabry病肾损害的临床、病理及基因突变特点的分析

目的 探讨Fabry病肾损害的临床病理及α-半乳糖苷酶A(α-Gal A)基因(GLA 基因)突变的特点.方法 回顾性分析14例Fabry病患者的临床、肾脏病理及GLA基因突变等特点.结果 Fabry病肾损害在肾活检患者中检出率为0.074％,平均确诊年龄(30.57±9.32)岁,男∶女=2.5∶1.尿蛋白量中位数为1.71 g/24 h[(0.32～ 4.71)g/24 h].5例有血尿,4例有肾功能受损,肾外受累的表现以血管角质瘤最多见(10/14),其次为心脏病变(6/14).经典型患者9例,迟发型5例,其中6例有肾脏病家族史.肾脏病理光镜下可见明显的肾小球细胞空泡变性,部分患者可见硬化的肾小球.电镜下2例女性患者为部分足细胞内有髓磷脂样小体形成,其余病例所有足细胞内均可见髓磷脂样小体.4例测定α-Gal A活性的先证者均低于正常值.12例先证者进行了GLA基因突变分析,11例发现有GLA基因突变.3个新突变为碱基插入或缺失突变,临床表型均为经典型Fabry病.大多数迟发型患者携带的基因突变位于酶结构的包埋区或部分包埋区(3/11).在已证实的GLA基因突变中,携带I91T、R112H、Q312H的先证者主要表现为“迟发型”;携带W162X、F169S、S201F、N272K及L310R的先证者均表现为“经典型”.结论 本组Fabry病肾损害患者占肾活检的0.074％,常伴有血管角质瘤及心脏受累,且不同的GLA基因突变可能与患者的表型密切相关.     

Identification of the CLCN7 gene mutations in two Chinese families with autosomal dominant osteopetrosis (type II)

Here we report the identification of two different mutations in chloride channel 7 gene in two unrelated patients with autosomal dominant osteopetrosis type II. We determined that one patient (a 32-year-old woman) carried a heterozygous gene for a R767W mutation in exon 24, and another patient (a 17-year-old boy) carried a heterozygous gene for a novel frameshift mutation (Glu798FS) in exon 25. Recent studies have reported loss-of-function mutations in the chloride channel 7 (CLCN7) gene as a cause of autosomal dominant osteopetrosis type II (ADO-II). The identification of gene mutations in Chinese with ADO has not been reported previously. In this study, we identified mutations of the CLCN7 gene in two unrelated Chinese families with ADO-II. Two probands with ADO-II were diagnosed based on their bone characteristics on X-rays and their laboratory results. All 25 exons of the CLCN7 gene, including the exon–intron boundaries, were sequenced. We found in family 1 that the proband (a 32-year-old woman) was heterozygous for a CLCN7 mutation. The nonsynonymous mutation consisted of a heterozygous C/T transition at codon 2327 in exon 24, which resulted in an arginine (CGG)-to tryptophan (TGG) substitution at position 767 (R767W). The same heterozygous mutation (C/T) was determined in her father and son, who were asymptomatic with normal skeleton radiography. In family 2, we found that the proband (a 17-year-old boy) carried a novel frameshift mutation (Glu798FS) resulting from a G insertion between codon 60 and codon 61 in exon 25. The heterozygous –/G insertion is predicted to elongate the peptide of CLCN7 by 120 amino acids after position 797 amino acids. Similarly, some individuals of this family carried the same heterozygous mutation, but they are all asymptomatic. Furthermore, the R767W and Glu798FS mutations were not found in 100 unrelated controls. Our present findings suggest that the novel Glu798FS mutation in exon 25 and R767W in exon 24 in the CLCN7 gene were responsible for ADO-II in these Chinese patients.     

Denys-Drash综合征及其致病基因突变鉴定二例报道

目的 研究中国Denys-Drash综合征(DDS)患者Wilms瘤基因1(WT1)的基因突变。 方法 应用PCR方法扩增出WT1基因全部10个外显子及其相邻内含子序列，经纯化后进行PCR产物直接测序。 结果 2例患者WT1基因分别存在1个杂合错义突变。例1的X外显子9第1180位碱基C→T突变，造成第394位精氨酸改变为色氨酸，即p.R394W(c.1180C>T)。例号2的S外显子9第1203位碱基C→A突变，造成第401位组氨酸改变为谷氨酰胺，即p.H401Q(c.1203C>A)。其中第1203位碱基C→A突变，p.H401Q (c.1203C>A)，在国内外文献及突变数据库中均未见报道，属新发现的突变。 结论 DDS综合征WT1基因中外显子9为突变热点，并发现一种新的WT1基因突变。     

Mutational analysis of the RET proto-oncogene in a kindred with multiple endocrine neoplasia type 2A and Hirschsprung's disease.

BACKGROUND/PURPOSE: Germline mutations of the RET proto-oncogene (RET; 10q11.2) have been reported in multiple endocrine neoplasia type 2A (MEN 2A) and Hirschsprung's disease. The authors investigated a Japanese kindred in which MEN 2A and Hirschsprung's disease frequently have been found. METHODS: The pedigree consisted of 28 members (11 boys and 17 girls) spanning 4 generations, of whom, 8 were affected with MEN 2A or Hirschsprung's disease. RESULTS: Direct sequence DNA analysis of the RET proto-oncogene showed a heterozygosity for a G to C transition at the second nucleotide of codon 620 (exon 10) in the patients, resulting in the replacement of cysteine by a serine residue in the affected Ret protein. This family added a novel RET missense mutation (C620S) predisposing to the association of MEN 2A and Hirschsprung's disease. CONCLUSION: Detection of the mutated RET gene carriers may be used for genetic counseling of potential risk for Hirschsprung's disease as well as MEN 2A in the affected families.     

A novel small insertion mutation, C.1030_1031ins (T) in α-galactosidase A leads to renal variant fabry disease

Fabry disease is a rare X-linked recessive glycosphingolipid storage disease that is caused by a deficiency of the lysosomal α-galactosidase A (GLA) enzyme, encoded by the GLA gene. This deficiency leads to the accumulation of glycosphingolipids throughout the body, which, in turn, causes multisystem diseases associated with renal, cardiovascular, and cerebrovascular complications. Recent molecular studies of GLA have demonstrated the existence of atypical variants in Fabry disease, suggesting significant genotype-phenotype correlations. In this study, we describe a renal variant of Fabry disease caused by a novel small insertion mutation in the GLA gene.     

A novel missense mutation, Leu390Val, in the cardiac beta-myosin heavy chain associated with pronounced septal hypertrophy in two families with hypertrophic cardiomyopathy

OBJECTIVE: An examination of the genetic background and phenotypic presentation of familial hypertrophic cardiomyopathy (FHC) with respect to specific mutations in the MYH7-gene encoding the cardiac beta-myosin heavy chain. SETTINGS: Two families (n = 22) from a cohort of 67 families with FHC were studied at the National University Hospital, Rigshospitalet, Copenhagen. METHODS: Clinical, non-invasive examinations of all included family members followed by molecular genetic analysis including PCR-single strand conformation polymorphism/heteroduplex (SSCP/HD) analysis and sequencing of exon 3-23 of the MYH7-gene. RESULTS: We found FHC associated with a missense mutation in two families, i.e. a C > G transversion at position g10124 and a G > T transversion at position g10126 causing the change of a leucine residue at codon 390 to a valine residue. The mutation is located in the actin-binding region of the beta-myosin heavy chain. The leucine residue is evolutionarily conserved in vertebrate myosins. In the two families, the phenotypic presentations in the clinically affected were characterized by asymmetric septal hypertrophy (septum diameter 18.8 (5.0) mm (mean (SD)) with only minor involvement of the left ventricular free wall (posterior wall diameter 11.0 (2.2) mm). Furthermore, the left ventricular systolic and diastolic functions were well preserved, even at a high age. The symptomatic status of the clinically affected patients depended on the presence or absence of a concomitant left ventricular outflow tract gradient. CONCLUSIONS: We report a novel missense mutation associated with FHC caused by a double nucleotide transversion. The penetrance of the mutation was not complete, but in clinically affected patients the mutation gives rise to an echocardiographic phenotype, predominantly characterized by pronounced septal hypertrophy.     

A novel uromodulin mutation in autosomal dominant tubulointerstitial kidney disease: a pedigree-based study and literature review

Autosomal dominant tubulointerstitial kidney disease caused by mutations in uromodulin gene (ADTKD-UMOD) is a spectrum of hereditary renal disorders, characterized by early-onset hyperuricemia, gout and progressive nephropathy. This study presented a novel UMOD mutation in an ADTKD pedigree and reviewed studies in Chinese population. The index patient is a 16-year-old girl with hypertension, hyperuricemia and normal serum creatinine level. Four affected and six unaffected members were available for genetic screen. The mutation analysis was performed by next-generation sequencing and direct sequencing. A literature research was conducted to review Chinese ADTKD-UMOD cases. MEDLINE and Chinese Biomedicine Databases were searched with ‘uromodulin’, ‘juvenile gout’ and their related terms. Genetic sequencing revealed a de novo mutation within exon 3 (Cys223Gly), which was co-segregating with phenotype in this pedigree. In the review, four studies and our study involving a total of 67 ADTKD patients from 11 families were identified. Of these patients, 27 were confirmed to carry UMOD mutations. Mutations occurred in exon 3 were commonly observed, while mutations within exon 4, 5 and 9 occurred less frequently in Chinese ADTKD-UMOD cases. Among these cases, median age of symptom onset was 26.5?years, median age of end-stage renal diseases (ESRD) or death by ESRD was 41.9?years without renal replacement treatment. Phenotype caused by mutations in D8C domain seemed to be severe than those in GPI domain. Compared with patients of other race, Chinese ADTKD-UMOD patients advanced more aggressively to ESRD.