应用PCR技术对DMD患者进行缺失检测与产前基因诊断

运用聚合酶链式反应(polymerasechainreaction,PCR)技术对3个Duchenne型肌营养不良症(DMD)家系中的患者进行dystrophin基因内9个外显子缺失检测,在2个家系中检测到外显子45、48、51缺失,同时运用PCR技术扩增位于dystrophin基因内内含子短串联重复序列,对非缺失型DMD家系进行了产前诊断,胎儿为正常女性.dystrophin基因外显子缺失检测方法快速、敏感、准确,可在临床推广中应用;短串联重复序列(STR)多态性分析方法可用于DMD家系的产前基因诊断和携带者检出.     

Structural analysis of DMD gene and its clinical application in Chinese.I.Bgl Ⅱ exon—containing fragment,RFLP and carrier detection

This article is one of the serial studies on the characteristics of the molecular structure for dystrophin gene in Chinese.By using the entire dystrophin cDNA(14kb) as a probe,the number and RFLPs of Bgl Ⅱ exon-containing fragments of the dystrophin gene were analysed.Four new Bgl Ⅱ fragments were found,two of them(3.7 and 6.2 kb) detected by comparing the hybridization patterns with cDNA1-2a,1a and 2a,one(9.3 kb) from the hybridization pattern with cDNA 9 by lengthening migrating distance of DNA fragments in electrophoresis,and another and (4.0 kb) by comparing the patterns with cDNA 11-14, 11a,11b,aac-12a and 14.The results indicated that the number of Bgl Ⅱ exon-containing fragments should be 59 rather than 55 reported previously,which laid the foundation of the Bgl Ⅱ partial restriction map for dystrophin gene.Three of the four RFLPs found in Caucacian appear in the hybridization patterns of three subclones,i.e. cDNA 2b-3,cDNA 4-5,and cDNA 5b-7.The values of expected heterozygote frequency(EHF) were 0.33,0.33 and 0.40,and the observed heterozygote frequency(OHF) were 0.40,0.40 and 0.48 respectively.Meanwhile,two new rare allelic fragments(15kb) were found in RFLPs from Bgl Ⅱ/2b-3 and Bgl Ⅱ/4-5a patterns respectively.These Bgl Ⅱ RFLPs and four XbaI RFLPs documented in our laboratory have been used to detect the carrier in 7 DMD families and 1 BMD family.Of the 69 individuals from the 8 families,11 females were diagnosed as the carriers with DMD mutation,4 females as the doubtful carriers,12 females were defined as normal genotype and 2 females as probably normal.The results suggest that the carrier testing method based on dosage intensity analysis and genotype analysis by using dystrophin cDNA as a probe will be more sensitive and accurate.     

一例X染色体结构重排导致DMD疾病

假肥大型肌营养不良(Duchenne/Becker muscular dystrophy,DMD/BMD)是一种最常见的进行性肌营养不良疾病，呈X-连锁隐性遗传，主要由DMD基因的缺失、重复及点突变所致，极少数病例是由于染色体结构重排破坏了DMD基因而引起疾病的发生。本文报告了1例经多重连接探针扩增技术(multiplex ligation-dependent probe amplification,MLPA)和下一代测序检测后原因未明的、具有典型症状的DMD患者。采用核型分析、FISH分析及三代测序、 Sanger测序综合分析发现，患者存在母源性的X染色体臂间倒位(Chr.X:g.[31939463–31939465del; 31939466–131765063 inv; 131765064–131765067del])半合子变异。由于该变异破坏了DMD基因和HS6ST2基因，因此推测该变异是患者发病的遗传学病因。患者表现肌无力等典型的DMD症状，没有明显的Paganini-Miozzo综合征相关症状。本病例的明确诊断，提示结构重排破坏DMD基因也是导致DMD重要原因之一；常规遗传学...     

山东省假性肥大型肌营养不良的RFLP分析 Ⅰ．可疑携带者的基因型确定

本文选用10个DMD基因内部和桥探针,对山东省9个地区的21个DMD／BMD家系的173名成员进行RFLP分析,其中可疑携带者55人,多数家系应用1－3个探针,有基因重组家庭选用4－5个探针,便可完成多态分析,RFLP分析可以确定85．45％的可疑携带者（≥95％可信限）,即13人确定为携带者,30人排除携带者,另有4人风险大幅提高,通过这些探针的群体多态检测和不同家庭结构和类型的应用分析,我们提出了在中国人群中DMD／SMD基因诊断的基本程序。     

正常中国人和DMD患者X染色体Xp~(21)区的RFLP研究

本文应用从人类X柒色体Xp~(21)区不同部位分离得到的9种DNA探针,分析了100名正常中国人,38名DMD患者及其母亲X柒色体Xp~(21)区的14个限制性位点多态性(RSP;又称限制性片段长度多态性,RFLP)。发现正常的X染色体与携带DMD基因的X染色体Xp~(21)区的RFLP频率没有明显差别;在38例DMD患者中有7例的X染色体有DNA片段缺失;在本文分析的24例患者母杀中有17例是DMD基因携带者,她们在Xp~(21)区的RFLP均存在杂合的多态性,因此可以应用RFLP连锁分析对这些家系进行DMD的产前诊断。     

Identification of binding epitope of a monoclonal antibody (Z12) against human TNF-α using computer modeling and deletion mutant technique

Humantumornecrosisfactor-a(hTNF-a)isanunglycosylated,pleiotropiccytokinewithnumerousbiologicaleffectsincludingcytotoxicandproinflam-matoryactivities[1].Asrevealedbytheresearchwithmurinemodel,hTNF-playedakeyroleinmanydis-easessuchasrheumatoidarthritis,multiplesclerosisandinflammatoryboweldisease[2],andtherefore,be-cameausefultargetoftherapyforthediseases.Neu-tralizingmonoclonalantibody(mAb)againsthTNF-,asanagentblockinghTNF-activity,hasbeenusedfortherapyofthosediseasesabove[3,4].ThemAb,de…     

Preliminary study on the molecular structure of 3'''' region of Duchenne muscular dystrophy (DMD) gene in Chinese

Number and order of HindⅢ exon-containing fragments (Hd) at 3' region of DMD gene were studied systematically using 16 partly-overlapping cDNA subprobes which were produced from dystrophin cDNA 9- 14 with each of 9 restriction endonudeases. There are 25 Hd fragments corresponding to cDNA 9 -14 in DMD gene. Since then, the exact length and the new order of Hd fragments are established. A new 2.1 kb fragment (Hd 55) is revealed, a 5.2 kb fragment (formely designated as Hd 59) is excluded and the existence of a controversial 3.2 kb fragment (Hd 64) is confirmed. Besides, three new exons were revealed by comparing the PvuⅡ and the XbaⅠ hybridization patterns with the Hindlll hybridization patterns for these cDNA subprobes. It is concluded that there are at least 66 Hd fragments, or 79 exons in DMD gene basing on the discovery of three additional exons. The corresponding relationship between the 66 Hd fragments and the SfiⅠ large scale physical map has been studied, and at least 17 Hd fragments or 19 exo     

Integrated Genetics Inc.宣布将推出几种新的DNA试验法

新近出现了杜兴病(DMD)的产前和病原携带者检测的有效方法。现在每350名男性中就有1人可能患此病,是最常见的遗传性肌肉疾病。DMD是一种有严重危险的肌肉疾病,危害骨骼肌、呼吸肌和心肌。DMD基因(人类最大的已知基因之一)位于X染色体上,且在所有携带缺陷型基因的男性体内表达。症状的发作一般出现在出生后2～5年,而在出生后20～30年时致死。过去五年间,强化了对该基因的鉴定和搞清DMD的     

Expression of estrogen receptor (ER) -α and -βtranscripts in the neonatal and adult rat cerebral cortex, cerebellum, and olfactory bulb

INTRODUCTIONEstrogen has been known to exert extensive effects via estrogen receptor (ER) on diverse physio-logical and develoPmental functions of the brain[1,2]. It has been observed that the distribution ofthe classical ER subtype-a (ERa) and the recentlycharacterized novel ER subtype--fi (ERg), and theirexpression patterns (ERcr/ERfi) vary greatly amongvarious brain regions[1, 3]. These evidences suggestthat each ER subtype may play a different role inestrogen's effects on the br…     

PRMT2β suppresses autophagy and glycolysis pathway in human breast cancer MCF-7 cell lines

Autophagy as a novel therapeutic target can inhibit or increase treatment efficacy in various types of breast cancer in a cell-type-dependent manner [1,2].Several studies have revealed that the coordination between Akt and the glycolytic pathway plays an indispensable role in mediating autophagy and caspase-dependent apoptosis,suggesting that a new regulatory mechanism for the process [3,4].Protein arginine N-methyltransferases(PRMTs)are eukaryotic enzymes that catalyze the transfer of methyl groups from S-adenosylmethionine to arginine residues of numerous PRMT substrates [5,6].PRMT2(also known as HRMT1L1)belongs to the arginine methyltransferase family [7].PRMT2β is a novel PRMT2 splice variant isolated from breast cancer cell [8].It occurs at the 3′ end of the PRMT2,resulting in loss of exons 7–9 and downstream frame-shifting [9].PRMT2β possesses 83 new amino acids at the C-terminus and its size is 301 amino acids.Our previous study reported that PRMT2β has potential antitumor effect by suppressing cyclin D1 expression [10].However,little is known about whether PRMT2β could regulate autophagy and glycolysis of MCF-7 cells.     

人基因组YAC分子克隆库的构建及DMD基因YAC克隆的筛选

以pYAC4为载体,以正常人白细胞和含4条X染色体的细胞株GM1414为DNA源构建成人基因组YAC(Yeast Artificial Chromosome,酵母人工染色体)分子克隆库,已得到原始克隆近2万个,插入DNA片段长度在400—1000kb,从其中选出一组YAC克隆,它们含有DMD基因全部DNA顺序。     

Identification and expression of epidermal growth factor gene in mouse testis

INTRODUCTIONEpidermalgrowthfactor(EGF)wasinitiallyisolatedandpurifiedfromthesubmaxillarygland(SMG)ofmalemouse[1].Itisapolypeptidecomposedof53aminoacidresidues[2].Itinfluencescellproliferationanddifferentiationandmodulatesthegrowthanddevelopmentofmammalianorgans[3--7].AnoteworthyfindingisthatextirpationofmouseSMGresultsinamarkedreductionofserumEGFconcentrationassociatedwithanimpairedspermatogenesis[3].ThisfindingsuggeststhatEGFmayregulatespermproductionanddifferentiation.Inhumantest…     

Peptide 68-88 of apocytochrome c plays a crucial role in its insertion into membrane and binding to mitochondria

Apocytochromec(Apocyt.c)istheheme-freeprecursorofcytochromec.Itissynthesizedinthecytosolandpost-translationallyimportedintomitochondriabyfollowingaquiteuniquepath-waycomparedtoothermitochondrialprecursorproteins[1].ItdoesnotpossessacleavableN-terminalpresequence,andneitheramembranepotentialnorATPisrequiredforitsimport[2].Further,noproteinaceouscomponentresponsibleforitsimporthaseverbeenidentified[3,4].Intheintermembranespaceapocyt.cisconvertedintoholocytochromecbycytochromechemelyase(CCHL)…     

Functional expression of opioid receptor—like receptor and its endogenous specific agonist nociceptin／orphanin FQ during mouse embryogenesis

INTRODUCTIONOpioidreceptorsbelongtotheG-protein-coupledreceptorfamilythatischaracterizedbytheseventransmembranespanningdomainsinstructure.Threesubtypesoftheopioidreceptors(H,6,andK)havebeenclonedandcharacterizedthroughtheir1.CorrespondingauthorFunctionalexpressionofORLIandN/OFQduringmouseembryogenesisdistinctaffinitiesfordifferentopioidligands.TheseopioidreceptorsareallcoupledtotheinhibitoryGprotein(Gi)andnegativelyregulateadenylatecyclase[1].Opioidreceptor--likereceptor(ORLI),anew…     

Nor1, a novel cytokine-responsive regulator of pancreatic β-cell mass and function mediated by disruption of mitochondrial networks

Type 2 diabetes(T2D)is a chronic metabolic disease characterized by insulin resistance and hyperglycemia,which is ultimately linked to the loss of pancreaticβ-cells and their function[1].Understanding the pathological mechanisms ofβ-cell dysfunction in T2D may lead to development of new therapeutic approaches.Recently,compelling evidence suggests that members of the nuclear receptor 4A(NR4A)subgroup play a pivotal role inβ-cell loss[2].Nor1,also known as NR4A3,belongs to the NR4A subfamily,which also includes Nur77(NR4A1)and Nurr1(NR4A2),and is defined as a true orphan nuclear receptor with an unknown endogenous ligand or ligand independent[3].As a regulator of gene expression located in the nucleus,Nor1 exhibits tissue-specific expression,which selectively controls diverse biological processes,including cell proliferation,apoptosis,differentiation,immune homeostasis,and fuel utilization[4].Thus far,it was reported that Nor1 is involved in numerous pathologies such as cancer,inflammatory diseases,and Parkinson’s disease[4].     

NO Level and Endothelial NO Synthase Gene Polymorphism （Glu298Asp） in the Patients with Coronary Artery Disease from the Turkish Population

A healthy endothelium plays a core role in cardiovascu-lar control [1]. In the endothelial cell, nitric oxide (NO) issynthesized by the endothelial nitric oxide synthase (eNOS)encoded by a 26-exon gene (NOS 3) located on chromo-some 7 [2]. Besides its regulatory functions on vasomotortone and blood flow, endothelial NO is known to inhibitthe platelet activation and modulate migration and growthof the vascular smooth muscle [3]. Indirect evidence sug-gests that alterations of the NO pathwa…     

拟澳洲赤眼蜂和短管赤眼蜂在米蛾多卵上的种间竞争

1引言 小菜蛾(Plutella xylostella)是十字花科蔬菜的一种重要害虫.20世纪70年代以来,化学杀虫剂的大量使用和抗性的产生.使小菜蛾成为南亚、东南亚和中国南方蔬菜生产中最为严重的害虫[8,22].因此,寻求安全有效的生物防治措施是防治小菜蛾的必要途径[23].赤眼蜂(Trichogramma)是一类多食性的卵寄生蜂,近百年来已被成功地用于防治为害多种作物及森林的害虫[21],但研究和利用赤眼蜂防治小菜蛾只有10来年的历史[12,23].     

The binding of actin to p38 MAPK and inhibiting its kinase activity in vitro

Ineukaryocyte,themitogen-activatedproteinkinases(MAPKs)playcriticalrolesinmanysignaltransductionprocesses[1].p38signalpathwayisanimportantbranchoftheMAPKs[2].Oneoftheprimaryfunctionsofp38medicatestheinflammatorysignalbyphosphorylatingATF[3].Itisknownthatinhibitingthep38activitycanblockthesignaltransductionofinflammationandsub-sequentlyalleviateinflammatoryresponse[4].Inrecentyears,severalresearchgroupshavetriedtousesomespecificinhibitorsofp38forclinictrial[5—7].IthasbeendemonstratedthatP…     

Inhibition of invasiveness and expression of epidermal growth factor receptor in human colorectal carcinoma cells induced by retinoic acid

INTRODUCTIONAsaderivativeofvitaminA,RAcaninhibittheproliferati0nofmanymalignantcel1sallde1icitdifferentiationinsometumorce1lsI1-5].RecentstudieshaveshownthatRAmodu1atessynthesisofover4Oproteinsthroughitsnucleicrecept0r[6].Forinstance,RAinducesthesynthesis0ffibronectin(FN)incertaintumorcellsandactivatesthegenecodingfOrB1sllbunitoflaminin(LN)causingitsexpressi0n[7,8].FNandLN'areknownt0bethemostimportantcomponentsofnon-c0llageng1ycoproteinsintheextracellu1armatrixandareclose1yrelatedt…