基于HRM方法筛选乳腺癌组织MYBL2基因突变热点区域

MYB家族成员——MYBL2基因,是一个重要的原癌基因,可通过多种途径抑制细胞凋亡并参与细胞周期调控,但其在乳腺癌发生和发展中的作用尚不明确.本试验通过采用高分辨率熔解曲线法(HRM),结合DNA测序分析检测MYBL2基因第5、8、13和14外显子在乳腺癌组织中的突变情况.结果显示有12例(约占32%)乳腺癌组织在MYBL2基因第8号外显子第1 472位存在A→G的突变,导致编码氨基酸序列发生S→G的突变;而外显子5、13和14未检测到突变.结果提示MYBL2基因的8号外显子的突变可能与乳腺癌的形成和发展有密切联系.     

全外显子测序结合先天性心脏病相关基因过滤在一家族性房间隔缺损家系中检测出新的致病突变TBX20(D176N)(英文)

研究目的:寻求该房间隔缺损家系遗传致病原因。创新要点:1.鉴定出一个全新的家族性房间隔缺损相关性TBX20突变;2.首次使用全外显子测序结合先天性心脏病相关基因过滤的方法来研究小家系遗传致病因素;3.TBX20的T-box DNA结合域的突变与先天性心脏病有关。研究方法:对一个临床发现的房间隔缺损家系(图1a)的先证者进行全外显子测序,运用公共数据库过滤后,使用先天性心脏病相关基因再次过滤,得到了19个候选基因;然后,运用SIFT、Polyphen-2和MutationTaster等软件预测,排除了13个多态性位点(表2);最后,运用共分离检测(聚合酶链式反应产物直接测序),找到该家系致病的遗传因素,即TBX20基因发生了错义突变(D176N)(图2),该突变位点在ESP和dbSNP数据库中也未曾发现,且该位点在多种生物中高度保守(图1c)。重要结论:1.本研究发现的TBX20突变(D176N)是该房间隔缺损家系致病的原因,同时该突变位点为世界上首次报道;2.全外显子测序结合先天性心脏病相关基因过滤是一个分析小家系遗传致病因素的有效又经济的方法。     

一个中国Lynch综合征家系携带的MLH1基因第19号外显子移码突变:一项家系研究（英文）

目的:寻找一个Lynch综合征患者所在家系携带的DNA错配修复基因突变,探讨各突变对肿瘤发生发展的影响。创新点:MLH1的第19号外显子c.2250_2251insAA移码突变既往被认为是意义未名突变,而我们的研究为明确该突变的致病意义提供了依据。另外,我们首次报道了MLH3基因第1号外显子c.1397C>A突变。该突变有可能使Lynch综合征患者的发病年龄提前。方法:运用免疫组织化学技术检测家系先证者肿瘤组织中错配修复基因蛋白的缺失情况,使用二代测序技术通过先证者血标本明确患者所携带的突变。同时运用Sanger法检测家系其他成员该突变的携带情况以明确突变对肿瘤发生发展的影响。结论:我们在患者体内发现MLH1基因第19号外显子移码突变(c.2250_2251insAA)以及MLH3基因第1号外显子c.1397C>A突变。在患者家系中,我们仅检测到有MLH1突变,因此该突变极有可能为致病突变。     

全外显子组测序发现一个中国Nance-Horan综合征家系NItS基因的新突变

研究目的：通过对一个中国Nance-Horan综合征家系的临床表型及基因突变分析,揭示本家系的致病遗传机制。研究方法：对该Nance—Horan综合征家系的一个男性患者进行全外显子组测序,结合此家系临床表型及遗传方式分析,选定x染色体上NHS基凶上的一个无义突变c．322G〉T（E108x）为可疑致病突变。通过聚合酶链式反应（PCR）和Sanger测序,对该家系内其他成员进行NHS基因突变分析,同时对50名健康对照者的NHS基因的突变检测结果进行对比。另外,将该突变的位点第108位氨基酸残基进行多物种NHS蛋白内序列比对。最后,对该家系成员眼部及全身的临床特点进行全面检查和分析。重要结论：全外显子组测序结合Sanger测序发现NHS基因第一个外显子上的c．322G〉T（E108X）突变为引起该家系临床病变的突变位点;多物种NHS蛋白内序列比对发现该突变位点第108位氨基酸残基位于高度保守区;临床表型分析发现该家系内存在表型异质性。此家系为国内首次报道的无义突变引起的Nance-Horan综合征家系。     

全外显子组测序发现一个中国Nance-Horan综合征家系NHS基因的新突变(英文)

研究目的:通过对一个中国Nance-Horan综合征家系的临床表型及基因突变分析,揭示本家系的致病遗传机制。研究方法:对该Nance-Horan综合征家系的一个男性患者进行全外显子组测序,结合此家系临床表型及遗传方式分析,选定X染色体上NHS基因上的一个无义突变c.322GT(E108X)为可疑致病突变。通过聚合酶链式反应(PCR)和Sanger测序,对该家系内其他成员进行NHS基因突变分析,同时对50名健康对照者的NHS基因的突变检测结果进行对比。另外,将该突变的位点第108位氨基酸残基进行多物种NHS蛋白内序列比对。最后,对该家系成员眼部及全身的临床特点进行全面检查和分析。重要结论:全外显子组测序结合Sanger测序发现NHS基因第一个外显子上的c.322GT(E108X)突变为引起该家系临床病变的突变位点;多物种NHS蛋白内序列比对发现该突变位点第108位氨基酸残基位于高度保守区;临床表型分析发现该家系内存在表型异质性。此家系为国内首次报道的无义突变引起的Nance-Horan综合征家系。     

一种新的PITX2基因缺失/插入移码突变引起的Axenfeld-Rieger综合征研究(英文)

研究目的:对1个Axenfeld-Rieger综合征家系的临床特点及基因突变进行研究,探索Axenfeld-Rieger综合征发病的遗传机制。研究方法:对该Axenfeld-Rieger综合征家系进行全面临床检查,对家系成员应用聚合酶链反应(PCR)扩增PITX2基因和FOXC1基因的所有外显子及相邻内含子,对其产物进行直接测序并对PITX2基因第5个外显子进行克隆测序。选取100名健康者作为对照组,应用PCR扩增PITX2基因第5个外显子并进行直接测序。应用SWISS-MODEL软件对野生型和突变型的PITX2蛋白同源域进行建模分析。重要结论:该Axenfeld-Rieger综合征家系的眼部表型多样,但是各患者的全身系统异常却呈现一致性(见图2;表1)。基因测序结果显示先证者及其他患者均具有PITX2基因杂合突变c.198_201delins TTTCT(p.M66Ifs*133)。尽管PITX2基因突变引起Axenfeld-Rieger综合征已经被广泛证实,但是PITX2基因缺失/插入移码突变引起的Axenfeld-Rieger综合征仅被报道过一次,我们的研究首次在中国人群中揭示了这种罕见的突变方式。     

Amazing Letters

ATTITUDE=100%Sample Text如果令:A B C D E F G H I J K L M N O P Q R S T U V W X Y Z分别等于百分之1234567891011121314151617181920212223242526那么Hard work(努力工作)H A R D W O R K=8 1 18 4 23 15 18 11=98%Knowledge(知识)K N O W L E D G E=11 14 15 23 12 5 4 7     

莆田地区非小细胞肺癌患者EGFR基因突变分析

分析莆田地区非小细胞肺癌(NSCLC)患者表皮生长因子受体(EGFR)基因突变的情况。收集138例莆田地区非小细胞肺癌组织,采用EliVisionTM plus免疫组织化学染色检测癌组织中EGFR基因外显子18、19、20及2l的突变,同时分析其突变与临床特征的关系。结果:138例NSCLC中共检出52例EGFR基因突变,EGFR突变阳性率为37.7%;外显子19和21突变占总突变的92.3%;腺癌突变发生率占突变总数的73.1%;女性EGFR基因突变率(55.0%)显著高于男性(30.6%)(P<0.05)。结果表明:莆田地区NSCLC患者EGFR基因突变以外显子19和21突变为主,女性患者和腺癌患者是选用EGFR酪氨酸激酶抑制剂的优势人群。     

家族性肥厚型心肌病相关cTnI基因突变检测芯片的应用

基于基因芯片技术对心肌钙蛋白(cTnI)基因上的突变进行了分析.针对外显子上的突变特征设计了特异性探针,制备了可以同时检测cTnI基因上第3,5,7,8外显子突变的低密度基因芯片.对每一个外显子,设计了2条5'端标记荧光的寡核苷酸链,一条模拟野生型序列,另一条模拟突变型序列,将二者混合起来模拟杂合子序列.经过实验条件的优化,制作的芯片可以灵敏、特异地检测cTnI基因外显子上的突变.结果表明,该芯片检测突变正配错配区别明显,荧光强度比值符合理论估计(正中碱基错配的信号强度是完全正配信号强度的50%左右).经进一步对芯片优化后,该芯片有望在家族性肥厚型心肌病的研究和诊断中得到应用.     

水牛IGF1基因多态性与其生长性状、血清浓度以及IGF1和IGF1R基因表达的相关性研究（英文）

目的:探讨胰岛素样生长因子1(insulin-like growth factor1,IGF1)基因单核苷酸多态性(SNP)与埃及水牛生长性状的关联性。创新点:发现IGF1基因多态性对埃及水牛的生长性状、血液生化指标和基因表达有显著影响,并为埃及水牛的选育提供重要的分子遗传标记。方法:以200头埃及水牛为试验对象,对其IGF1基因的多态性进行检测,并分析其与生长性状、血液生理生化指标和基因表达的相关性。结论:在水牛IGF1基因多态性分析中,发现两个新的SNP位点(G64A和G280A)分别分布在外显子1和外显子4的非编码区。不同基因型的统计分析表明,G64A和G280A位点的GG基因型水牛个体分别在3~6月龄和6~9月龄的体重(BW)和平均日增重(ADG)均显著高于其他基因型(P0.05)。这两个SNPs位点的组合基因型产生了三种单倍体GG/GG、AG/AG和AA/AA。在3~12月龄的水牛个体中,单倍体基因型与BW和ADG存在显著关联(P0.0001)。纯合的GG/GG单倍体基因型水牛生长性能优于其他水牛。两个SNP位点与半腱肌中IGF1和IGF1R的mRNA水平以及IGF1血清浓度水平相关。此外,GG/GG单倍体水牛表现出较高的mRNA和血清浓度水平。综上所述,这两种SNP位点G64A和G280A可作为埃及水牛生长性状选育的重要分子遗传标记。     

Polymorphisms in the genes for coagulation factor II,V,VII in patients undergoing coronary angiography

INTRODUCTIONCoronaryarterydisease (CAD)continuestobethemajorcauseofmorbidityandmortalityinourcountry .TheactivityofbloodcoagulationfactorshasbeenshowntobeanimportantriskindicatorforCAD .However,circulatinglevelsofcoagulationfactorsmaynotaccuratelyreflectt…     

Polymorphisms in the genes for coagulation factor II,V, VII in patients undergoing coronary angiography

Objective: To determine whether polymorphisms in the genes for coagulation factor. II, V, VII could predispose an individual to increase risk for coronary artery disease (CAD) and/or myocardial infarction (MI) in Chinese. Methods: We screened coagulation factor II(G20210A), V(G1691A), VII (R353Q and HVR4) genotype in 374 patients undergoing coronary angiography by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) assay. Results: The R353Q and HVR4 gemotype of the factor VII distribution was in accordance with Hardy-Weinberg equilibrium. The frequencies of FVII genotype or allele did not show statistically significant differences between CAD group and controls or between male and female. The frequencies of the Q allele and (RQ+QQ) genotype were significantly higher among the CAD patients without myocardial infarction (MI) history than among those with MI history (P<0.05). However, HVR4 polymorphism was not significantly different within groups. We only find one normal control of factorII (G20210A) mutation. No coagulation factor V(G1691A) mutation was found in the CAD patients and controls. Conclusion: The factor II(G20210A), V(G1691A) mutation is absent and may not be a major genetic factor for CAD and/or MI; the Q allele of the R353 Q polymorphism of the factor VII gene may be a protective genetic factor against myocardial infarction in Chinese. Project (No. 021103166) supported by a grant from the Key Project of Science and Technology Commission of Zhejiang Province, China     

A new mutation （1062 del 16） of iduronate-2-sulfatase gene from a Chinese patient with Hunter syndrome

Objective: To identify the mutations of iduronate-2-sulfatase (IDS) gene, to reveal its mutation features, and to establish a basis for genetic counseling and prenatal gene diagnosis of Hunter syndrome. Methods: Urine glycosaminoglycans (GAGs) assay, PCR and DNA sequencing were performed to detect mutation of IDS gene of the patient and his parents. Results: The result showed that the patient was: DS( ), HS( ), KS(-), CS(-), and that both of his parents were negative. A frame-shift deletion mutation (1062 del 16) was identified in exon 7 of the patient's IDS gene. His parents' genotypes were normal. Conclusion:The patient's mutation was not inherited by his parents but a novel one. The mutation probably altered the primary structure and tertiary structure of IDS enzyme protein remarkably and lowered the activity of IDS enzyme greatly. Therefore it is supposed to be the direct cause of the disorder.     

Genotype-phenotype correlations in Chinese patients with TGFBI gene-linked corneal dystrophy

In this paper, we report the clinical and molecular features of the distinct TGFBI (human transforming growth factor β-induced, OMIM No. 601692) gene-linked corneal dystrophy. Altogether, five pedigrees and ten unrelated individuals diagnosed as corneal dystrophy were recruited. Peripheral venous DNA was extracted, and then amplified by polymerase chain reaction (PCR) and scanned for mutation by single-stranded conformation polymorphism (SSCP). Direct DNA sequencing was used to analyze the mutations of the TGFBI gene. In our study, thirty patients from five pedigrees and ten sporadic patients were diagnosed as four TGFBI gene-linked corneal dystrophies of granular corneal dystrophy type I (GGCD I), Avellino corneal dystrophy (ACD), lattice corneal dystrophy type I (LCD I), and lattice corneal dystrophy type IIIA (LCD IIIA), and in total, seven disease-causing mutations, namely R555W, A546D, A546T, and T538P mutations in exon 12, R124H and R124C mutations in exon 4, and P501T mutation in exon 11, were identified, while four polymorphisms of V327V, L472L, F540F, and 1665-1666insC were screened in exons 8,11, and 12. The study ascertained the tight genotype-phenotype relationship and confirmed the clinical and genetic features of four TGFBI gene-linked corneal dystrophies.     

Novel mutation c.980_983delATTA compound with c.986C>A mutation of the FRMD7 gene in a Chinese family with X-linked idiopathic congenital nystagmus

Objective

To screen mutations in FERM domain-containing protein 7 (FRMD7) gene in two Chinese families with X-linked idiopathic congenital nystagmus (XLICN).

Methods

Common ophthalmic data and peripheral blood of two Chinese XLICN families (families A and B) were collected after informed consent. Genomic DNA was prepared from the peripheral blood of members of the two families and from 100 normal controls. Mutations in the FRMD7 gene were determined by directly sequencing polymerase chain reaction (PCR) products.

Results

We identified a novel mutation c.980_983delATTA compound with c.986C>A mutation in the 11th exon of FRMD7 in family B, and a previously reported splicing mutation c.782G>C (p.R261G) in family A. The mutations were detected in patients and female carriers, while they were absent in other relatives or in the 100 normal controls.

Conclusions

Our results expand the spectrum of FRMD7 mutations in association with XLICN, and further confirm that the mutations of FRMD7 are the underlying molecular mechanism for XLICN.     

Deletion analysis of SMN1 and NAIP genes in southern Chinese children with spinal muscular atrophy

Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of lower motor neurons and occasionally bulbar motor neurons leading to progressive limb and trunk paralysis as well as muscular atrophy. Three types of SMA are rec-ognized depending on the age of onset, the maximum muscular activity achieved, and survivorship: SMA1, SMA2, and SMA3. The survival of motor neuron (SMN) gene has been identified as an SMA determining gene, whereas the neuronal apoptosis inhibitory protein (NAIP) gene is considered to be a modifying factor of the severity of SMA. The main objective of this study was to analyze the deletion of SMN1 and NAIP genes in southern Chinese children with SMA. Here, polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) was performed to detect the deletion of both exon 7 and exon 8 of SMNI and exon 5 of NAIP in 62 southern Chinese children with strongly suspected clinical symptoms of SMA. All the 32 SMAI patients and 76% (13/17) of SMA2 patients showed homozygous deletions for exon 7 and exon 8, and all the 13 SMA3 patients showed single deletion of SMN1 exon 7 along with 24% (4/17) of SMA2 patients. Eleven out of 32 (34%) SMA1 patients showed NAIP deletion, and none of SMA2 and SMA3 patients was found to have NAIP deletion. The findings of homozygous deletions of exon 7 and/or exon 8 of SMN1 gene confirmed the diagnosis of SMA, and suggested that the deletion of SMN1 exon 7 is a major cause of SMA in southern Chinese children, and that the NA1P gene may be a modifying factor for disease severity of SMA 1. The molecular diagnosis system based on PCR-RFLP analysis can conveniently be applied in the clinical testing, genetic counseling, prenatal diagnosis and preimplantation genetic diagnosis of SMA.     

Deletion analysis of SMN1 and NAIP genes in southern Chinese children with spinal muscular atrophy

Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of lower motor neurons and occasionally bulbar motor neurons leading to progressive limb and trunk paralysis as well as muscular atrophy. Three types of SMA are recognized depending on the age of onset, the maximum muscular activity achieved, and survivorship: SMA1, SMA2, and SMA3. The survival of motor neuron (SMN) gene has been identified as an SMA determining gene, whereas the neuronal apoptosis inhibitory protein (NAIP) gene is considered to be a modifying factor of the severity of SMA. The main objective of this study was to analyze the deletion of SMN1 and NAIP genes in southern Chinese children with SMA. Here, polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) was performed to detect the deletion of both exon 7 and exon 8 of SMN1 and exon 5 of NAIP in 62 southern Chinese children with strongly suspected clinical symptoms of SMA. All the 32 SMA1 patients and 76% (13/17) of SMA2 patients showed homozygous deletions for exon 7 and exon 8, and all the 13 SMA3 patients showed single deletion of SMN1 exon 7 along with 24% (4/17) of SMA2 patients. Eleven out of 32 (34%) SMA1 patients showed NAIP deletion, and none of SMA2 and SMA3 patients was found to have NAIP deletion. The findings of homozygous deletions of exon 7 and/or exon 8 of SMN1 gene confirmed the diagnosis of SMA, and suggested that the deletion of SMN1 exon 7 is a major cause of SMA in southern Chinese children, and that the NAIP gene may be a modifying factor for disease severity of SMA1. The molecular diagnosis system based on PCR-RFLP analysis can conveniently be applied in the clinical testing, genetic counseling, prenatal diagnosis and preimplantation genetic diagnosis of SMA. Project supported by the National Natural Science Foundation of China (No. J0710043), and the Natural Science Foundation of Zhejiang Province (No. 2007C33049), China     

Identification of <Emphasis Type="Italic">FECH</Emphasis> gene multiple variations in two Chinese patients with erythropoietic protoporphyria and a review

Erythropoietic protoporphyria (EPP), an autosomal dominant disease, is caused by partial deficiency of ferrochelatase (FECH), which catalyzes the terminal step of heme biosynthesis because of loss-of-function mutations in the FECH gene. To date, only a few cases have been described in Asia. In this study, we describe the clinical features of two Chinese patients with EPP, with diagnosis confirmed by the increase of free protoporphyrin in erythrocytes, detection of plasma fluorescence peak at 630–634 nm, and analysis of FECH gene mutations. Using gene scanning, we identified a small deletion in the FECH gene (c.973 delA) in one proband (patient A) and a pathogenic FECH mutation (c.1232 G>T) in the other (patient B) and also observed some nucleotide variations (c.798 C>G, c.921 A>G, IVS1?23 C>T, IVS3+23 A>G, IVS9+35 C>T, and IVS3?48 T>C) in these patients. The family pedigree of patient A was then established by characterization of the genotype of the patient’s relatives. We also analyzed the potential perniciousness of the missense mutation with bioinformatic software, Polyphen and Sift. In summary, Chinese EPP patients have similar manifestations to those of Caucasians, and identification of the Chinese FECH gene mutations expands the FECH genotypic spectrum and may contribute to genetic counseling.