泛发性肥大细胞增多症一例及KIT基因突变研究

目的 检测1例泛发性肥大细胞增多症患者KIT基因突变情况,为患者提供预后判断及治疗选择。 方法 收集患者临床资料,提取患者,其父母以及200例健康人外周血DNA,采用PCR扩增KIT基因编码区的全部外显子及其侧翼序列并测序。 结果 基因检测发现患者外周血DNA中的KIT基因发生c.1526A > T杂合突变,导致氨基酸出现p.Lys509Ile改变,父母及200例健康对照均未见相同突变。 结论 KIT基因p.Lys509Ile突变可能为泛发性肥大细胞增多症患者病因之一。     

Sjogren-Larsson综合征ALDH3A2基因突变研究

目的 检测2例以先天性鱼鳞病、智力发育迟缓及痉挛性截瘫为主要表现的Sj?觟gren-Larsson综合征患者的ALDH3A2基因突变情况。 方法　2例分别为2岁女性及1.5岁男性儿童,家族中均无类似疾病。提取2例患者及其6例相关亲属（每例患儿的父母及哥哥）外周血DNA,采用PCR扩增ALDH3A2基因编码区的全部外显子及其侧翼序列并测序。同时随机抽取100例无关健康人外周血基因组DNA作对照。结果基因检测发现,例1的ALDH3A2基因发生c.325G>A纯合突变,导致氨基酸出现p. Gly109Arg改变,其父母及其未受累的兄长为该突变的杂合携带者。例2的ALDH3A2基因发生c.1157A>G及c.1294A>T复合杂合突变,导致氨基酸出现p. Asn386Ser及p.Arg432X改变,突变分别来自父母。100例健康对照者均未见相同突变。结论 在2例Sj?觟gren-Larsson综合征患者中检测到ALDH3A2基因的p. Gly109Arg纯合突变及p. Asn386Ser与p. Arg432X复合杂合突变,该基因突变可能与患者临床表型相关。 【关键词】 Sjogren-Larsson综合征; 鱼鳞病; 突变; ALDH3A2     

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。     

基因检测确诊表现为多发性咖啡斑的1型神经纤维瘤病2例

目的对2例表现为多发性牛奶咖啡斑的散发患儿进行神经纤维瘤病致病基因检测,早期确定其诊断。方法提取患儿及其父母外周血DNA,采用PCR扩增先证者NF1基因所有外显子及其侧翼序列并测序,并以200例无关正常人作为对照。结果 2例散发患儿除多发性咖啡斑以外均无神经纤维瘤病的其他表现。我们在患者1和患者2基因组DNA中发现NF1基因分别发生c.4600CT杂合无义突变(p.R1534X)以及c.7348CT杂合无义突变(p.R2450X)。这两个突变位点既往均未见报道。2例患儿的父母及200例健康正常人均未检测到相应突变位点。结论 2例多发性咖啡斑患儿经基因诊断均确诊为1型神经纤维瘤病,NF1基因的p.R1534X及p.R2450X突变分别为其致病突变。基因诊断是早期确诊神经纤维瘤病的有效方法。     

长岛型掌跖角化病二例SERPINB7基因突变研究

目的 报告2例长岛型掌跖角化病,确定其致病基因突变。 方法 收集患者及其父母外周血和临床资料,提取基因组DNA,PCR扩增SERPINB7基因8个外显子及其侧翼序列,对扩增产物进行DNA测序以查找基因突变位点,并以200例无关健康人DNA作为对照进行扩增测序。 结果 2例患者均存在SERPINB7基因c.796C > T纯合突变,导致编码蛋白质第266位氨基酸出现终止改变（p.R266*）,其父母均为c.796C > T杂合突变,而无关健康对照未发现上述突变。 结论 SERPINB7基因的c.796C > T突变可能是引起2例患者长岛型掌跖角化病的原因。     

Netherton综合征一例SPINK5基因突变研究

目的 检测Netherton综合征患者SPINK5基因的突变情况。 方法 收集患者临床资料,提取患者及其相关亲属外周血DNA,用PCR扩增SPINK5基因编码区的全部外显子及其侧翼序列并测序。 结果 直接测序发现患者SPINK5基因的第13号外显子中的第1111位碱基发生C→T杂合突变（c.1111C > T）,导致其编码第371位氨基酸变为终止密码子（p.R371X）;第32号外显子中的第3121位碱基发生C→T杂合突变（c.3121C > T）,导致其编码第1041位氨基酸发生错义突变（p.R1041C）,其健康父母为相应突变的杂合携带者,200例健康对照未见该突变。 结论 SPINK5基因的p.R371X及p.R1041C复合杂合突变可能是引起该患者临床表现的病因之一。     

先天性大疱性鱼鳞病样红皮病角蛋白10的新发突变

目的: 检测1例先天性大疱性鱼鳞病样红皮病患者KRT1和KRT10基因突变.方法: 提取患者及其家人外周血DNA,PCR扩增KRT1和KRT10基因编码区的全部外显子及其侧翼序列并测序,以100名正常人作对照.结果: 该患者KRT10基因第1号外显子中的第466位碱基发生C→T杂合突变(c.C466T),导致其编码第156位氨基酸发生错义突变(p.R156C),患者父母、妹妹及正常对照均未发现该突变,提示其为新发突变.结论: KRT10基因的c.C466T错义突变可能为引起该患者临床表型的病因.     

Carvajal综合征一例伴桥粒斑蛋白基因新突变

【摘要】 目的 对1例临床表现为羊毛状发,膝盖、掌跖角化性皮损,暂无心脏症状的患儿进行基因突变检测。方法 收集患儿及其父母的临床资料。提取患儿、其父母及100例无关健康对照者外周血DNA,采用二代皮肤靶向测序包检测患儿的基因突变,应用Sanger测序法进行验证。结果 患儿女,3岁,出生头发卷曲,8月龄出现掌跖角化并渐累及膝盖,其父母表型正常。测序发现,患儿桥粒斑蛋白（DSP）基因第23号外显子存在移码突变c.5152dupT（p.L1718Ffs*15）,DSP基因第24号外显子检测到无义突变c.C6478T（p.R2160X）。其母亲DSP基因第23号外显子亦存在c.5152dupT移码突变,但第24号外显子未检测到相关突变。其父亲及100例健康对照中均未检测到相关突变。诊断：Carvajal综合征。结论 该例Carvajal综合征患儿存在DSP基因复合杂合突变c.5152dupT（p.L1718Ffs*15）和c.C6478T（p.R2160X）,可能与其发病有关。     

TSC2基因发生体细胞镶嵌突变导致结节性硬化症一例北大核心CSCD

目的报告1例TSC2基因发生体细胞镶嵌突变导致结节性硬化症患者的临床特点和基因突变情况。方法采集1例临床拟诊断结节性硬化症患者及其父母外周血提取基因组DNA,采用聚合酶链反应（PCR）及第二代测序法扩增TSC1和TSC2基因的全部外显子及其侧翼序列,并进行DNA测序分析,查找基因突变位点,以200例无关健康人DNA作为对照。提取患者皮损处皮肤组织DNA,利用PCR扩增TSC2基因目的片段并测序。结果患者临床出现面部血管纤维瘤、腰部色素减退斑、甲周纤维瘤及肾血管平滑肌脂肪瘤,符合结节性硬化症的诊断标准。患者TSC2基因存在c．5130_5131insT突变（p．V1711Cfs＊18）,且瘤体DNA较外周血DNA携带更高频率突变。患者父母、无关正常对照及公开数据库均未发现该突变。结论TSC2基因发生体细胞镶嵌突变c．5130_5131insT是导致该患者较轻结节性硬化症表型的原因。     

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。     

痣样基底细胞癌综合征一家系PTCH1基因突变分析

目的 对痣样基底细胞癌综合征一家系进行PTCH1基因突变分析。 方法 提取先证者（Ⅱ5）及Ⅱ1、Ⅱ3、Ⅲ4的DNA,以50例健康人为对照。应用聚合酶链反应（PCR）、DNA直接测序明确突变位点,根据突变位点设计特异性引物,用PCR来检测突变位点从而进一步确定该家系的致病原因。 结果 先证者PTCH1基因的1条等位基因第14号外显子上2137位胞嘧啶C被胸腺嘧啶T替代（c.2137C > T）,即CAG→TAG,导致终止密码产生（Q714X）,Ⅲ4也检测到相同突变。健康对照者中未检出该突变。 结论 PTCH1基因的无义突变（c.2137C > T）可能是引起该患者临床症状的特异性突变。     

UV-specific p53 and PTCH mutations in sporadic basal cell carcinoma of sun-exposed skin

UVB irradiation is known to produce DNA damage at mutation hotspots in the p53 tumor suppressor gene, leading to the development of skin cancers. Mutations in the PTCH tumor suppressor gene, which is known to be responsible for the development of nevoid basal cell carcinoma syndrome, have also been identified in sporadic basal cell carcinomas (BCCs). We describe the case of an 80-year-old welder in whom 3 novel p53 mutations, as well as UV-specific PTCH mutations, were detected in two BCC samples from sun-exposed skin. The simultaneous presence of UV-specific p53 and PTCH mutations in the same BCC sample has not previously been reported.     

Clinical and genetic study in 22 patients with basal cell nevus syndrome

BACKGROUND: Nevoid basal cell carcinoma syndrome is an autosomal dominant disorder characterized by developmental abnormalities and cancer predisposition. The PTCH 1 gene, the human homolog of the Drosophila segment polarity gene patched, has been shown to be involved in the development of nevoid basal cell carcinoma syndrome. PTCH 1 is mapped to chromosome 9q22.3. The aim of the present study was to report on clinical and genetic characteristics in patients followed for nevoid basal cell carcinoma syndrome and to compare them to the data in the literature. PATIENTS AND METHODS: Screening for PTCH 1 mutations was done in 22 patients followed between 1981 and 2003 for clinical suspicion of nevoid basal cell carcinoma syndrome. Clinical and radiological data were reviewed retrospectively from records. Genetic analysis was performed using blood samples after patient informed consent was obtained. When possible, DNA was also analyzed from the parents of patients in whom PTCH 1 mutations were found. RESULTS: All patients had developed basal cell carcinomas: 45% palmar and plantar pitting, 62% jaw cysts and 66% calcification of falx cerebri. Medulloblastomas and meningiomas were the most common associated tumors. PTCH 1 mutations were identified in 13 patients: 6 familial cases, 3 sporadic cases and for 4 patients, it was not possible to conclude. Nine different new germ-line mutations were identified. DISCUSSION: Genetic analysis allows molecular confirmation of diagnosis in about half of all patients. Early diagnosis is essential for detection of clinical and radiological manifestations in young patients and for provision of advice concerning protection of the skin from the sunlight.     

Congenital linear unilateral basal cell nevus: a case report with patched gene molecular studies

BACKGROUND: Linear unilateral basal cell nevus represents a linear collection of macules and papules histologically similar to basal cell carcinoma but with benign clinical behavior. We describe a patient who initially presented at the age of 6 months with a unilateral linear basal cell nevus on the right flank. The differential diagnosis included the nevoid basal cell carcinoma syndrome. Constitutional PTCH mutations are causative of the nevoid basal cell carcinoma syndrome, and somatic PTCH mutations are found in the vast majority of basal cell carcinomas. Somatic SMO mutations have also been found in some basal cell carcinomas. METHODS: Histologic examination of the lesions is performed. Short tandem-repeat molecular analysis at the PTCH locus and sequencing of PTCH and SMO genes is performed. RESULTS: Histologic examination revealed features initially indistinguishable from basal cell carcinoma. Short tandem-repeat DNA analysis did not reveal loss of heterozygosity at the PTCH locus. DNA sequencing of both the PTCH and the SMO genes from the patient's lesions revealed neither inactivating mutations of PTCH nor activating mutations of SMO. CONCLUSION: Molecular examination indicates that the PTCH and SMO genes are not involved in the pathogenesis of the patients' congenital linear unilateral basal cell nevus. Furthermore, we discuss the relationship between linear basal cell nevus and basaloid follicular hamartoma.     

遗传性对称性色素异常症一家系ADAR基因检测

目的：检测遗传性对称性色素异常症一家系的ADAR基因突变。方法：提取家系中患者、健康成员及无血缘健康对照人群外周血样DNA,PCR扩增ADAR基因外显子后测序。结果：该家系中患者均存在ADAR基因第2号外显子,第982位碱基突变（c.982C>T,p.R328X）,突变导致第328位的精氨酸被终止密码替代。家系中健康成员及健康对照人群未发现该突变。结论：该遗传性对称性色素异常症家系患者中的ADAR基因突变（c.982C>T,p.R328X）可能与发病有关。     

Spectrum of PTCH1 mutations in French patients with Gorlin syndrome

Gorlin syndrome or nevoid basal cell carcinoma syndrome is an autosomal dominant disease characterized by developmental abnormalities and a predisposition to cancers. The responsible gene for this syndrome is the PTCH tumor suppressor gene encoding for the Sonic Hedgehog receptor. We screened for PTCH mutations in 65 French Gorlin syndrome families or sporadic cases for the first time. Nineteen novel mutations and five new polymorphisms were identified in this group of patients. One microdeletion without frameshift underlines the importance of one amino acid for Ptc receptor function. Although no mutation hot-spot was described, we identified a recurrent mutation.     

表皮松解性掌跖角皮症一家系KRT9基因新突变

目的:检测表皮松解性掌跖角皮症一家系患者角蛋白9(KRT9)基因突变。方法:收集家系成员的临床资料和血样,提取家系中4例患者和3名正常人及50名与本家系无关的正常对照外周血DNA,采用PCR技术扩增KRT9基因所有编码区并进行测序,分别检测家系中的突变情况。结果:该家系中所有患者均存在KRT9基因错义突变(c.484TC),导致第162位密码子由TCT(丝氨酸)转变为CCT(脯氨酸)(p.S162P),家系中3名正常个体和50名健康对照均未发现上述突变。结论:KRT9基因c.484TC错义突变是导致该家系发生表皮松解性掌跖角皮症的遗传基础。