杂合子导致CADASIL病患者家系NOTCH3基因突变

目的 研究一伴皮质下梗死和白质脑病的常染色体显性遗传性脑动脉病（CADASIL）患者家系NOTCH3基因突变,探讨突变分析方法在遗传性CADASIL疾病筛查和诊断中的应用。方法 收集该家系8位成员（5例患者,3例正常个体）外周血标本,提取基因组DNA。采用PCR扩增NOTCH3基因突变热点区域,DNA直接测序检测扩增产物,寻找该家系致病的突变基因。结果 NOTCH3基因第4外显子内存在一杂合的错义突变（c.421C＞T）,导致141位精氨酸（Arg）被半胱氨酸（Cys）替代。该家系中患者均携带该突变基因,而家系中正常个体未发现此突变基因。结论 杂合的错义突变（c.421C＞T）与该CADASIL家系中患者表型共分离,为引起该家系的致病基因突变。     

一个遗传性血管水肿家系C1抑制物基因突变的检测分析

目的 对一个遗传性血管水肿(HAE)家系患者C1抑制物(C1 INH)基因的突变类型进行检测分析。方法 用聚合酶链反应扩增产物直接测序法检测HAE患者C1 INH基因8个外显子及旁侧内含子序列,将检测结果与GenBank公布的C1 INH基因序列相比较,确定突变。为除外多态性可能,在30名正常人中对该突变进行分析。结果 该家系中的5例患者外显子8中均检测到1种新的突变类型(核苷酸序列17839 del C),正常人中无此改变。结论 在该家系中发现C1 INH基因核苷酸序列17839 del C突变,该突变可能是此家系发病的分子基础。     

A SPG3A mutation with a novel foot phenotype of hereditary spastic paraplegia in a Chinese Han family

Hereditary spastic paraplegia （HSP） （MIM#182600） is a group of heterogeneous neurodegenerative disorders, with 35 underlying loci recognized by the HGNC （HUGO Gene Nomenclature Committee; http：//www.gene.ucl.ac.uk/nomenclature/） and 10 identified genes （ http ： //www. gene. ucl. ac. uk/cgi-bin/nomenc lature/ searchgenes.pl plus NIPA1, last search July 2006）. The mode of inheritance may be autosomal dominant, autosomal recessive or X-linked. Among these, autosomal dominant spastic paraplegia （AD-HSP） is the most common type, accounting for 70%-80% of all families. The disease is characterized by lower limb spasticity, hyperreflexia, progressive spastic gait and an extensor plantar response. Pes cavus is one of the commonly reported foot phenotypes.     

家族性良性慢性天疱疮一家系及ATP2C1基因突变分析

目的 报道家族性良性慢性天疱疮一家系,并对ATP2C1基因突变进行分析。方法 收集家族性良性慢性天疱疮家系成员的一般资料,进行临床调查,绘制家系图谱。采用PCR及Sanger直接测序法对该家系中4例患者的ATP2C1基因进行检测,并以该家系3例健康者和100例无亲缘关系的正常人作为对照。结果 4例家族性良性慢性天疱疮患者ATP2C1基因的第21号外显子的第472位核苷酸由G变为A,该突变导致原先158位的天冬氨酸变为天冬酰胺（p.Asp158Asn）的错义突变。而家系中健康对照及无亲缘关系的正常人均未发现该突变。结论 ATP2C1基因的第21号外显子的第472位核苷酸由G变为A是ATP2C1基因新的突变位点,可能是家族性良性慢性天疱疮家系发病的主要原因。     

成骨不全家系一个新的Ⅰ型胶原α1链蛋白基因突变

目的 对Ⅰ型胶原α1链蛋白基因（COL1A1基因）进行测序研究，旨在寻找已知或未知的COL1A1基因突变位点，探讨我国成骨不全的发病机制。方法 研究一常染色体显性遗传成骨不全家系的临床特征，设计引物对家系中患者和正常人的COL1A1基因外显子进行扩增和测序分析，同时对群体中无血缘关系的50名健康对照者进行限制性内切酶分析。结果 发现家系中成骨不全患者COL1A1基因的第3470位点的碱基G→A的突变，导致G1157D，而在家系内非患者及正常对照者中均无发现。结论 COL1A1基因突变也是中国人群中成骨不全致病原因之一，现发现的突变属成骨不全一个新的致病基因突变。甘氨酸转变成天冬氨酸的这种突变对成骨不全表型具有重要的影响。     

家族性慢性良性天疱疮一家系致病基因新突变的发现

目的对家族性慢性良性天疱疮一家系的ATP2C1基因突变进行检测。方法采用聚合酶链反应扩增该家系患者和健康对照个体ATP2C1基因的全部外显子,直接测序法进行DNA测序,100例无亲缘关系的正常人作为对照。结果该家系患者ATP2C1基因内含子3的末端235-2碱基发生了1个腺嘌呤（A）→鸟嘌呤（G）的杂合性剪接位点突变。家系中健康对照个体及无亲缘关系的正常对照均未发现该突变。结论该剪接位点突变可影响基因转录和翻译产物,是ATP2C1基因新的特异性突变。     

Identification of a new lamin A/C mutation in a chinese family affected with atrioventricular block as the prominent phenotype

Even though mutations in LMNA have been reported in patients with typical dilated cardio-myopathy(DCM)and atrioventricular block(AVB)previously,the purpose of this study was to disclose this novel genetic abnormality in one Chinese family with the atypical phenotype of progressive AVB followed by DCM with normal QRS interval.Genome-wide linkage analysis mapped the AVB gene in this family to a marker at chromosome 1q21.2,where the LMNA gene was located.Direct DNA sequence analysis revealed a heterozygous G t...     

A novel deletion-frameshift mutation in the S1 region of HERG gene in a Chinese family with long QT syndrome

Background The congenital Long QT syndrome (LQTS) is a hereditary cardiac channelopathy that is characterized by a prolonged QT interval,syncope,ventricular arrhythmias,and sudden death.The chromosome ...     

一个汉族肥厚型心肌病家系中首次发现肌球连接蛋白-C基因Arg346fs突变

目的研究中国人家族性肥厚型心肌病(HCM)的致病基因突变位点,分析基因型与临床表型的相互关系。方法对5个经过MYH7基因扫描未发现异常的家族性HCM的先证者进行肌球连接蛋白-C基因(MYBPC3)扫描,聚合酶链反应(PCR)扩增其功能区外显子片断,双脱氧末端终止法测序。对阳性结果者进行家系中其他成员筛查,并分析患者临床表型特点。结果在1个家系中发现MYBPC3基因的13号外显子的Arg346fs突变,而正常对照组同一位置未见异常,Arg346fs突变为我国患者中首次发现。结论MYBPC3基因为我国家族性HCM的的致病基因之一。其临床表型的异质性提示多因素参与了HCM的发生及外显。     

Early-onset primary torsional dystonia in a 4-generation Chinese family with a mutation in the DYT1 gene

Primary torsion dystonia (PTD) is a clinically and genetically heterogeneous movement disorder. At least thirteen different types of dystonia can be distinguished on a genetic basis. The DYT1 gene was first mapped by Ozelius et al in 1989. Kramer et al linked the same locus to PTD in 12 Ashkenazi Jewish families in 1990.     

用位点特异PCR法检测Leber遗传性视神经病家系的mtDNA11778G→A点突变

目的 寻找一种快速、准确、有定量突变比例的简单PCR方法,以识别Leber遗传性视神经病(LH0N)息者所携带的mtDNA ll778G→4A点突变。方法 根据已知致病突变的碱基变化,设计M(突变)和N(正常)引物,分别与反向引物R配对扩增,严格控制退火温度和其他PCR反应条件达到特异扩增。病例组为LH0N家系共10个个体,对照组为40位正常人。结果 在先证者、母系已发病亲属和一个10岁男孩(未发病)体内分别检出突变比例各不相同11778G→A点突变,而在家系的正常配偶、父系子女及40位正常对照组未检出该突变。结论 位点特异PCR是一种不受DNA序列有否限制性内切酶位点的检测突变的方法,适用于LH0N等致病突变明确的遗传病的基因诊断。     

Exercise test on the patients with normokalaemic periodic paralysis from a Chinese family with a mutation in the SCN4A gene

Background Normokalaemic periodic paralysis （normoKPP） is characterized by transient and recurrent myoasthenia, and some patients also show muscle stiffness induced by cold exposure （paramyotonia congenita, PMC）. It is caused by a mutation in the muscle voltage gated sodium channel alpha subunit （SCN4A） gene. Due to the diversity of the clinical manifestations of patients, it is difficult for clinicians to differentiate some of patients with atypical normoKPP from those who suffer from other periodic paralysis and nondystrophic myotonia. So far, for normoKPP there are almost no ways to assist definite diagnosis besides genetic screening. This research was designed to evaluate an exercise test （ET） in confirming the diagnosis of normoKPP and in assessing the therapeutic effectiveness of some drugs on this disease. Methods ET, described by McMains, was performed on six subjects from a Chinese family, including four patients with overlapping disease of normoKPP and PMC caused by a mutation of SCN4A Met1592Val that is identified by genetic analysis and two normal control members. The change of compound muscle action potential （CMAP） was recorded. Besides the family, two patients were also tested during treatments with acetazolamide. Results All patients showed a slight increase in CMAP immediately after exercise, followed by an abnormal gradual decline, which reached its nadir 25-30 minutes after exercise. CMAP amplitude dropped by more than 40% in patients but less than 23% in controls. In the patients who received treatment with acetazolamide, the change of CMAP amplitude was less than 28% and, at any fixed times, less than pretreatment values. Conclusions The ET may be used as a predictive, easy and reliable method of diagnosing normoKPP under conditions without genetic screening help, and is an objective way to evaluate the therapeutic effectiveness. According to different response patterns, the ET may also be helpful in reducing the scope of genetic screening.     

一个Paget骨病家系SQSTM1及TNFRSF11A基因突变分析

目的 研究一Paget骨病家系SQSTM1基因及TNFRSF11A基因突变情况。方法 收集一Paget骨病家系,家系成员外周血中提取基因组DNA,应用聚合酶链式反应、直接测序对该家系成员SQSTM1及TNFRSF11A基因外显子进行测序。测序结果与GenBank公布的SQSTM1和TNFRSF11A基因正常序列对比,寻找有无突变。结果 在该家系中未发现与Paget骨病共分离的致病基因突变,检测到14个已知的单核苷酸多态性。结论 该家系成员的发病情况与SQSTM1、TNFRSF11A基因中发现的SNP无相关性,排除其为该Paget骨病家系致病基因的可能性。     

A novel mutation, Y255X, of the ARSB gene in a Chinese family with mucopolysaccharidosis type Ⅵ

Mucopolysaccharidoses (MPS) are lysosomal storage diseases with defectivedegradation of glycosaminoglycans. Mucopolysaccharidosis type Ⅳ( MPS Ⅳ )or Maroteaux-Lamy syndrome is caused by defective arylsulfatase B in the lysosomes ( ARSB; Nacetylgalactosamine-4-sulfatase, EC 3.1.6. 12 ) . The clinical manifestations of MPS VI include coarse facial features, growth retardation, short stature, skeletal malformations, hepatosplenomegaly, corneal clouding,and cervical myelopathy. Heart failure is the usual cause of death in the second or third decade of life. Despite all the physical disabilities, the intellect of MPS Ⅳ patients is preserved.     

A novel missense mutation of the ubiquitin protein ligase E3A gene in a patient with Angelman syndrome

Background  Angelman syndrome (AS) is a neurogenetic disorder caused by an expression defect of the maternally inherited copy of ubiquitin protein ligase E3A (UBE3A) gene from chromosome 15. Although the most common genetic defects include maternal deletions of chromosome 15q11-13, paternal uniparental disomy and imprinting defect, mutations in the UBE3A gene have been identified in approximately 10% of AS patients.

Methods  A Chinese girl of 28 months presented clinical manifestation of AS. Genetic diagnosis and molecular genetic defects were studied by methylation-specific PCR (MS-PCR) and linkage analysis by short tandem repeat (STR). We further performed sequence analysis of all the coding exons and flanking sequences of the UBE3A gene. The novel mutation screening was also performed in 100 unrelated healthy individuals to exclude the possibility of identifying a polymorphism variation.

Results  The MS-PCR analysis of the patient showed biparental inheritance of chromosome 15 with a normal methylation pattern in the 15q11-q13 region. And STR analysis revealed that the patient also inherited biparental alleles for six microsatellites. A novel mutation, cDNA1199 C>A (p.P400H), in exon 9 of the maternal UBE3A gene, was identified in the patient. Meanwhile, the mutation was observed in the patient’s mother who had a normal phenotype.

Conclusions  It is necessary to perform the UBE3A gene mutation analysis in non-deletion/non-UPD/non-ID patients with AS. The clinical picture of the patient is concordant with that observed in previously reported AS patients with UBE3A mutation.

     

葡萄糖激酶W257R突变致青少年发病的成人型糖尿病一家系分析

青少年发病的成人型糖尿病（MODY）是一类以常染色体显性模式遗传的单基因疾病,临床表现以无症状、轻度空腹血糖升高为特征,很少出现糖尿病并发症。本文报道一例中国人群中葡萄糖激酶（GCK）基因新发W257R突变所致的MODY。在先证者及父亲、弟弟中均发现GCK基因（Chr744187343）第7号外显子的杂合突变c.769T>C（p.W257R）。该家系中W257R突变在中国人群中为首发。     

CACNA1A 基因的错义突变R1345Q导致一种新的共济失调伴随发作性全身震颤：临床特征、基因诊断 及治疗的家系分析

目的CACNA1A基因编码P/Q型钙离子通道的亚单位,它的突变至少造成3种等位基因病：发作性共济失调2型（EA-2）、家 族性偏瘫性偏头痛1型（FHM1)和小脑脊髓共济失调6型（SCA 6）。本研究对一例19岁男性的发作性全身震颤患者的临床表现、 基因分析结果和治疗效果进行研究。方法对病人及家系中有类似症状的成员进行专科查体;对先证者的DNA进行下一代测序 分析以寻找致病基因,并用Sanger 测序方法对家系成员进行基因变异的验证。结果神经专科查体显示患者共济失调体征,醉酒 步态,头和躯干震颤。家系中另4个成员的症状和体征较轻。基因检测发现先证者携带有CACNA1A基因的杂合错义突变（NM_ 001127221.1 c.4034G->A, p.R1345Q, exon 25）,为致病突变。家系中4个患病成员中也携带同样杂合突变。病人经醋甲唑胺治 疗后效果不佳,但钙离子通道阻断剂西比灵治疗效果良好。结论根据患者的临床表现、基因突变类型和治疗效果,我们认为患者 CACNA1A基因突变R1345Q所引起的疾病不属于EA2,FHM1,或SCA 6任何一种,而是一种新的伴有发作性震颤共济失调。     

2型糖尿病线粒体基因变异的研究

目的：探索中国人２型糖尿病与线粒体变异的关系。方法针对已经报道的线粒体基因的高发突变区（ｍｔ３１５３－３５５１）近４００ｂｐ的片段在中国人正常人群、２型糖尿病群体及１２个母系遗传的２型糖尿病系中进行聚合酶链反应－单链构象多态性（ｐｏｌｙｍｅｒａｓｅ ｃｈａｉｎ ｒｅａｃｔｉｏｎ－ｓｉｎｇｌｅ ｓｔｒａｎｄ ｃｏｎｆｏｒｍａｔｉｏｎ ｐｏｌｙｍｏｒｐｈｉｓｍ,ＰＣＲ－ＳＳＣＰ）研究,发现异常构象再进     

风湿性心脏瓣膜病合并心房颤动患者中DNMT3 A与Kv1.5蛋白的表达变化及相关性研究

目的：探讨风湿性心脏瓣膜病合并心房颤动( AF)患者DNA甲基转移酶3A(DNMT3A)与超速延迟整流性钾通道蛋白(Kv1．5)表达水平变化及其相关性研究。方法将20例风湿性心脏瓣膜病接受瓣膜置换手术患者于体外循环前切取的右心耳组织作为研究对象,根据AF的定义将标本分为AF组(n=10)和窦性心律(SR)组(n=10),分别应用免疫组化、Western blot 法检测心房肌组织中 DNMT3A 和Kv1.5蛋白表达水平变化。结果免疫组化检测显示AF组与SR组相比,DNMT3A蛋白表达增加(P<0．05);而Kv1.5蛋白表达减少(P<0．05)。同时,Western blot检测显示AF组与 SR 组相比,DNMT3A 蛋白表达增加(P <0．05),而Kv1.5蛋白表达减少(P<0．05)。相关性分析显示DNMT3A与Kv1．5之间存在一定负相关性(r =-0．64, P <0．01)。结论 DNMT3A与Kv1.5之间存在负相关性,提示DNMT3A可能参与调控Kv1．5通道蛋白的表达。     

研究STEMI患者RBP4、HbA1c与miR-1-3p检测水平的诊断价值及与病情关系

目的 研究ST段抬高型心肌梗死（ST-segment elevation myocardial infarction,STEMI）患者视黄醇结合蛋白4（retinol-binding protein 4,RBP4）、糖化血红蛋白（glycosylated hemoglobin,HbA1c）与微小RNA（microRNA,miRNA）-1-3p检测水平的诊断价值及与病情关系。 方法 选取河北省保定市第二医院收治的136例STEMI患者为STEMI组,根据SYNTAX评分分为轻度病变组（n=55）、中度病变组（n=43）、重度病变组（n=38）3个亚组,选取同期65例体检健康者为对照组。收集研究对象临床资料,采用酶联免疫吸附测定检测血清RBP4和HbA1c水平,实时荧光定量聚合酶链式反应检测血清miR-1-3p水平。Spearman相关系数分析STEMI患者血清RBP4、HbA1c和miR-1-3p水平与SYNTAX评分的相关性。多因素Logistic回归分析STEMI发生的危险因素。受试者工作特征曲线分析血清RBP4、HbA1c和miR-1-3p水平对STEMI的诊断价值。 结果 与对照组比较,STEMI组吸烟史比例和总胆固醇、三酰甘油、低密度脂蛋白胆固醇（low density lipoprotein cholesterol, LDL-C）、空腹血糖、C反应蛋白（C reactive protein,CRP）、RBP4、HbA1c、miR-1-3p水平升高,高密度脂蛋白胆固醇水平降低（t/Z=2.459~9.608,P均＜0.05）。轻度、中度、重度病变组血清RBP4、HbA1c和miR-1-3p水平依次升高（F=18.202~48.798,P均＜0.001）。STEMI患者血清RBP4、HbA1c和miR-1-3p水平与SYNTAX评分呈正相关（r=0.588~0.647,P均＜0.001）。吸烟史、CRP、LDL-C、RBP4、HbA1c、miR-1-3p可作为STEMI发生的独立危险因素（OR=1.037~1.646,P均＜0.05）。血清RBP4、HbA1c和miR-1-3p联合诊断STEMI的曲线下面积高于各指标单独诊断（Z=3.025~4.213,P均＜0.05）。 结论 STEMI患者血清RBP4、HbA1c和miR-1-3p水平升高,与冠状动脉病变程度正相关,可作为反映STEMI患者病变程度的参考指标。