板层状鱼鳞病两家系TGM1基因的突变

目的:检测2个板层状鱼鳞病家系TGM1基因的突变情况.方法:采用PCR-DNA直接测序方法明确两个家系TGM1基因突变位点,并以免疫组化的方法检测其中一家系患者皮肤中转谷氨酰胺酶的活性.结果:在1个家系的患者中发现TGM1基因第13号外显子第2060位碱基发生G→A纯合突变,使密码子CGT→CAT,导致R687H突变:在另一个家系的患者中发现TGM1基因第4号内含子第一个核苷酸发生G→T及第5号外显子第760位碱基发生G→A杂合突变,分别导致IVS4 1G＞T及D254N的复合杂合突变,其父母均为相应突变的携带者.在50名无关正常人中未发现相同突变.R687H纯合突变患者皮肤转谷氨酰胺酶的活性降低.结论:R687H纯合突变及D254N和IVS4 1G＞T复合杂合突变为新发现的TGM1基因突变位点,可能是引起两家系板层状鱼鳞病患者的病因.     

Rothmund-Thomson综合征一例RECQL4基因的突变分析

目的 检测1例Rothmund-Thomson 综合征患者及其父母的RECQL4基因突变情况。方法收集1例中国汉族Rothmund-Thomson 综合征患儿及其父母的外周血标本,提取其外周血DNA,采用PCR扩增RECQL4基因编码区的全部外显子,DNA测序仪直接测序,明确突变位点,并以同样方法检测30例无关正常人作对照。结果 患者RECQL4基因发生2处突变：剪接位点突变IVS11-1G→A和无义突变3401 A→T,两突变分别来自其父母。30例正常人对照组不存在此两种突变。结论 该患者存在RECQL4基因的剪接位点突变IVS11-1G > A和无义突变3401 A > T。     

隐性营养不良型大疱性表皮松解症3例患者的COL7A1基因突变检测

目的检测3例隐性营养不良型大疱性表皮松解症患者COL7A1基因突变位点。方法提取3例患者及其相关亲属外周血DNA,采用PCR扩增COL7A1基因编码区的全部外显子及其侧翼序列并测序。结果基因检测发现3例患者共有5个突变位点,包括2个错义突变(p.P2232L与p.G1531E),2个无义突变(p.R2492*与p.R2777*)和1个剪切位点突变(ds IVS6+2TC),其中p.P2232L,p.G1531E,ds IVS6+2TC这三个突变位点未曾报道。结论上述突变可能参与了隐性营养不良型大疱性表皮松解症的致病机制。     

以“湿疹”为突出表现的Wiskott-Aldrich综合征患者WASP基因突变的检测

目的探讨1例以"湿疹"为突出表现的Wiskott-Aldrich综合征(Wiskott-Aldrich syndrome,WAS)患儿WASP基因突变。方法收集患儿的临床资料及外周血,提取基因组DNA,PCR扩增致病基因WASP基因外显子及剪切位点序列,通过DNA测序查找基因突变。结果发现患儿WASP基因第8号内含子存在一个剪切位点突变(IVS8+1ga)。结论 IVS8+1ga为WASP基因突变热点,本例患儿WAS诊断明确。本例提示,湿疹患儿如合并血小板减少症临床上需警惕WAS可能。     

毛囊角化病患者ATP2A2基因新剪切位点突变的检测研究

目的:检测1例严重表型毛囊角化病患者ATP2A2基因的突变.方法:采用聚合酶链反应及直接测序法对该患者进行ATP2A2基因的突变位点检测,运用反转录技术检测突变导致的RNA水平的变化,同时对120名无血缘关系健康者作为对照进行测序验证.结果:通过筛查在患者ATP2A2基因中发现一个新的剪切位点突变IVS18+5G＞C,反转录分析证实该突变的发生导致ATP2A2基因转录后在其突变等位基因的18号和19号外显子之间插入了27个核苷酸.结论:该例毛囊角化病患者的检测结果进一步扩充了ATP2A2基因的突变库,并为阐明ATP2A2基因的突变导致该病发生的分子机制提供了帮助.     

一对先天性大疱性鱼鳞病样红皮病双胞胎患者表型与角蛋白1基因突变研究

目的 检测先天性大疱性鱼鳞病样红皮病双胞胎患者角蛋白1、10（KRT1、KRT10）基因突变情况,探讨致病基因与表型间的关系。方法 收集1对先天性大疱性鱼鳞病样红皮病双胞胎患者及家族成员的临床资料。提取该双胞胎患者及其兄、父、母的外周血DNA,PCR扩增KRT1和KRT10基因编码区全部外显子及其侧翼序列并测序,以100例健康人作为对照。结果 先证者男,11岁,全身皮肤反复起水疱、肥厚伴脱屑11年;其双胞胎弟弟有类似皮损。2例患者KRT1基因1号内含子第1位碱基发生突变（c.591 + 1G > A）,而家系中3例正常成员和无亲缘关系的100例健康对照均未发现该突变。结论 KRT1基因1号内含子第1位碱基突变（c.591 + 1G > A）可能为引起该双胞胎患者临床表型的病因。     

不伴套叠性脆发的2例Netherton综合征致病基因突变检测

目的:通过基因检测的方法,确诊2例不伴有套叠性脆发表现的Netherton综合征患者。方法:采集患者临床资料,进行皮损组织病理检查及毛发镜检,提取患者及其相关亲属外周血DNA,采用PCR扩增相应可疑致病基因编码区的全部外显子及其侧翼序列并测序。结果:2例患者毛发均未见明显形态学异常。基因检测发现例1及其受累的妹妹SPINK5基因发生c.2260AT纯合突变,导致氨基酸出现p.Lys754*改变,未受累的父母及兄长为该突变的杂合携带者;例2的SPINK5基因发生c.1432CT及c.1693-2del A复合杂合突变,导致氨基酸出现p.Gln478*及剪切位点改变,突变分别来自其健康父母。200例健康正常对照者均未见相同突变。患者CDSN、KRT1、KRT10、KRT2及DSG1基因均未见致病性突变。结论:通过基因检测确诊2例Netherton综合征患者,基因检测为确诊临床表现不典型Netherton综合征的重要方法。     

Kaposi肉瘤组织中lyn基因的表达分析

目的 探讨Peutz-Jeghers综合征2个家系及3例散发病例外周血及息肉组织STK11基因的突变。方法 PCR扩增STK11基因前8个外显子及侧翼序列,进行DNA直接测序。结果 家系1所有患者息肉组织STK11基因的外显子8检测到1个新杂合突变,第2039位碱基鸟嘌呤被胞嘧啶替代,导致308位氨基酸由色氨酸（W）转变为半胱氨酸（C）,即W308C错义突变;外周血未检测到此突变位点。家系1所有患者外周血STK11基因的内含子2第12703碱基检测到1个新突变,该处发生A→G替换突变;息肉组织未检测到此突变位点。家系2所有患者外周血STK11基因的内含子7第16215碱基存在G→C杂合突变,已被前人证实为SNP。3例散发病例中未检测到突变位点。结论 STK11基因G2039C突变可能是部分Peutz-Jeghers综合征患者发病的遗传基础。     

一个遗传性对称性色素异常症家系DSRAD基因剪接位点突变的鉴定

目的 鉴定遗传性对称性色素异常症家系DSRAD基因的突变。方法 收集遗传性对称性色素异常症1个家系成员的血样，提取基因组DNA，用PCR扩增结合直接测序的方法进行DSRAD基因的检测。在内含子区域检测到一个碱基替换后，进一步提取患者外周血RNA，行RT-PCR，直接测序后分析其异常剪接方式。结果 在该家系患者的11号内含子区域检测到1个非经典的剪接位点突变，（c.3021-2G > A），RT-PCR结果发现，12号外显子缺失，在13号外显子处发生移码突变。结论 DSRAD基因11号内含子区域剪接位点突变（c.3021-2G > A）造成mRNA的异常剪接，导致邻近的12号外显子缺失和13号外显子移码突变，从而引起该家系患者发病。     

罕见亚型大疱性表皮松解症3例及家系调查

【摘要】 目的 报道3例罕见亚型遗传性大疱性表皮松解症（EB）。方法 收集先证者及其亲属的临床资料,全外显子测序筛查先证者致病基因,采用Sanger或qPCR测序对患者及其亲属进行突变验证。结果 例1表现为背部线状红色瘢痕,患者及有相似临床表现的母亲、无症状女儿均携带COL7A1基因c.4573G>A（p.Gly1525Arg）突变。例2表现为全身网状色素沉着,偶伴手足水疱,携带KRT5基因c.74C>T（p.Pro25Leu）新发突变。例3表现为曝光部位为主的色素异常伴左手不完全并指,先证者携带FERMT1基因2-6号外显子纯合缺失突变,分别来自无症状父母。例1诊断为显性痒疹型营养不良型 EB, 例2诊断为斑驳色素型单纯型EB, 例3诊断为Kindler EB。结论 EB临床异质性高,基因检测对于罕见亚型EB的明确诊断非常重要。     

Novel and recurrent FERMT1 gene mutations in Kindler syndrome

Kindler syndrome (OMIM 173650) is an autosomal recessive condition characterized by skin blistering, skin atrophy, photosensitivity, colonic inflammation and mucosal stenosis. Fewer than 100 cases have been described in the literature. First reported in 1954, the molecular basis of Kindler syndrome was elucidated in 2003 with the discovery of FERMT1 (KIND1) loss-of-function mutations in affected individuals. The FERMT1 gene encodes kindlin-1 (also known as fermitin family homologue 1), a 77 kDa protein that localizes at focal adhesions, where it plays an important role in integrin signalling. In the current study, we describe five novel and three recurrent loss-of-function FERMT1 mutations in eight individuals with Kindler syndrome, and provide an overview of genotype-phenotype correlation in this disorder.     

Síndrome de Kindler,manejo multidisciplinario

Kindler syndrome is a very rare form of bullous epidermolysis. It is a hereditary condition caused by a mutation in the FERMT1 gene that encodes the protein kindlin-1. It is clinically characterized by trauma-induced blistering, diffuse skin atrophy, poikiloderma, pseudosyndactyly, and photosensitivity. The most common mucosal manifestations are conjunctivitis, ectropion, hemorrhagic gingivitis, periodontal disease, premature tooth loss, and severe colitis.We present the first 4 cases of Kindler syndrome diagnosed at the Instituto Nacional de Salud del Niño in Lima, Peru. These cases highlight the unique clinical presentation and multiple manifestations of this disease and show how a multidisciplinary management approach kept symptoms under control and significantly improved patient quality of life.     

Heterozygous HPS1 mutations in a case of Hermansky-Pudlak syndrome with giant melanosomes

We report a Japanese man with Hermansky-Pudlak syndrome, having oculocutaneous albinism with a bleeding diathesis. Gene analysis of the patient's peripheral blood cells revealed that he was a compound heterozygote for HPS1 gene mutations. One of the mutations was a novel frameshift mutation at codon 321 (a G insertion) in exon 11 ( approximately 962-963insG), and the other was a 5' splice-junction mutation of IVS5 (IVS5 + 5G-->A). The content of eumelanin in the patient's hairs was significantly reduced. Histological analysis using light and electron microscopy revealed that melanocytes in the patient's epidermis contained an appreciable number of giant melanosomes. Cultured melanocytes from the patient's skin also contained giant melanosomes. Our finding of mutations in the HPS1 gene in relation to abnormalities in melanosome morphology and melanin production shed light on the role and function of the HPS1 gene product in the synthesis of melanosomes and melanin pigment.     

Costello综合征合并皮肤松弛症一例基因突变检测

目的 报告1例并发皮肤松弛症的Costello综合征患者,并进行分子遗传学诊断.方法 收集1例合并有皮肤松弛症的Costello综合征患者临床资料,提取该患者皮肤组织和其父母及150例无关健康对照的外周血基因组DNA,对HRAS基因所有外显子和侧翼序列测序.结果 患儿女,13月龄,生长发育迟缓,重度营养不良,面容粗糙,四肢皮肤严重松弛,皮下脂肪减少甚至消失.患儿HRAS基因第2外显子出现突变c.34G＞ T(p.Gly 12Cys),但其父母及健康对照均未检测到该位点突变.结论 HRAS基因第2外显子c.34G＞ T(p.Gly12Cys)突变可能为该例Costello综合征的致病原因.     

Kindler syndrome with severe mucosal involvement in childhood

Kindler syndrome (KS) is an inherited dermatosis linked to the FERMT1 gene, and is characterized clinically by trauma‐induced acral skin blisters in infancy and childhood, photosensitivity, and progressive poikiloderma. We report a case of KS in a 7‐year‐old Indian girl with severe mucosal involvement of the oral cavity and genitourinary tract. Mutation analysis in the girl showed a homozygous FERMT1 mutation, c.862C>T, p.R288*. The clinical manifestations in patients with KS show significant inter individual variation, even with the same type of mutations and within members of the same family. Our case highlights the role of environmental modifiers in regulating the clinical features of KS.     

X性连锁少汗性外胚层发育不良家系ED1基因突变检测

目的 探讨X性连锁少汗性外胚层发育不良(XLHED)家系中ED1基因突变。方法 收集2个X性连锁少汗性外胚层发育不良家系外周血标本;采用聚合酶链反应(PCR)结合DNA直接双向测序的方法。结果 家系1中ED1基因的第8个外显子下游与内含子8交界处存在一个新的剪接点缺失突变(IVS8+5 del G)。家系2中第9个外显子处存在一个错义突变(A959G)。这些突变未在两个家系的正常人及188例无关正常对照者中出现。结论 中国人ED1基因突变可引起XLHED,且IVS8+5del G为一个新的突变。     

红细胞生成性原卟啉病一家系中亚铁螯合酶基因突变的检测

目的 对一红细胞生成性原卟啉病（EPP）家系进行基因突变研究,探讨基因突变与临床表现的关系,为进一步开展基因诊断和基因治疗奠定基础。方法 收集家系资料,抽取家系中患者、正常人及与该家系无关的50例正常人的外周血,提取外周血基因组DNA。应用PCR方法扩增外周血基因组DNA亚铁螯合酶（FECH）基因的第1至11外显子及其侧翼序列。PCR产物直接进行双向测序以检测突变。结果 根据临床表现和卟啉测定结果,患者明确诊断为红细胞生成性原卟啉病。PCR扩增得到预期DNA片段。PCR 产物直接测序结果：家系中先证者、其妹和其父FECH基因第1内含子供体剪接位点检测到一个杂合突变（IVS1 ＋ 1G→C）,该突变为国际首次报道。还在先证者、其妹和其母FECH基因第1内含子受体端检测到一个与低表达等位基因相关的多态性（IVS1-23C/T）。结论 报道一FECH基因第1内含子供体剪接位点的新突变,该突变可能引发FECH基因缺陷,是EPP家系中患者发病的分子基础。     

Recurrent mutations in kindlin-1, a novel keratinocyte focal contact protein, in the autosomal recessive skin fragility and photosensitivity disorder, Kindler syndrome

Kindler syndrome (OMIM 173650) is a rare autosomal recessive disorder characterized by trauma-induced blister formation (especially in childhood) and photosensitivity. Other features include mucocutaneous scarring and progressive poikiloderma. There is also an increased risk of skin and mucous membrane malignancy. The disorder was recently mapped to 20p12.3 and pathogenic mutations were identified in a new gene, KIND1. This gene encodes a 677 amino acid protein, kindlin-1, a component of focal contacts in keratinocytes. In this study, we identified four new recurrent mutations in KIND1 in 16 individuals with Kindler syndrome from 13 families of Pakistani (676insC), UK Caucasian (E304X), Omani (W616X), or Italian (958-1G > A) origins. Haplotype analysis demonstrated common ancestral mutant alleles for each mutation, apart from one of the six Pakistani families in which the mutation 676insC (which occurs in a repeat of seven cytosines) was present on a different genetic background. All mutations were homozygous, apart from the three UK Caucasian cases that were all compound heterozygotes (second allele mutations: L302X, 1161delA, 1909delA). All mutations were associated with markedly reduced or absent skin immunostaining with an antikindlin-1 antibody. These loss-of-function KIND1 mutations demonstrate the importance of kindlin-1 in maintaining epithelial integrity, although the mechanism linking this mutant protein to photosensitivity and poikiloderma remains to be determined. Delineation of these recurrent mutations is also relevant to optimizing mutation detection strategies in Kindler syndrome patients from particular ethnic backgrounds.     

Kindler syndrome: a focal adhesion genodermatosis

Kindler syndrome (OMIM 173650) is an autosomal recessive genodermatosis characterized by trauma-induced blistering, poikiloderma, skin atrophy, mucosal inflammation and varying degrees of photosensitivity. Although Kindler syndrome is classified as a subtype of epidermolysis bullosa, it has distinct clinicopathological and molecular abnormalities. The molecular pathology of Kindler syndrome involves loss-of-function mutations in a newly recognized actin cytoskeleton-associated protein, now known as fermitin family homologue 1, encoded by the gene FERMT1 . This protein mediates anchorage between the actin cytoskeleton and the extracellular matrix via focal adhesions, and thus the structural pathology differs from other forms of epidermolysis bullosa in which there is a disruption of the keratin intermediate filament–hemidesmosome network and the extracellular matrix. In the skin, fermitin family homologue 1 is mainly expressed in basal keratinocytes and binds to the cytoplasmic tails of β1 and β3 integrins as well as to fermitin family homologue 2 and filamin-binding LIM protein 1. It also plays a crucial role in keratinocyte migration, proliferation and adhesion. In this report, we review the clinical, cellular and molecular pathology of Kindler syndrome and discuss the role of fermitin family homologue 1 in keratinocyte biology.