CYP1A1基因多态性与宫颈鳞癌的关系

目的:检测宫颈鳞癌组织中CYP1A1等位基因的表达,探讨CYP1A1基因多态性在宫颈鳞癌发生中的作用。方法:收集宫颈鳞癌组织标本50例,以正常宫颈组织61例为阴性对照。采用等位基因特异性PCR法(ASA-PCR)和PCR扩增限制酶切法(RFLP-PCR)检测宫颈鳞癌组织中CYP1A1等位基因Exon7位点和MspⅠ位点多态性;用SPSS13.0软件建立数据库,进行χ2检验、logistic回归分析。结果:(1)CYP1A1Exon73种多态基因型在宫颈鳞癌组和对照组分布差异有统计学意义(P<0.005),宫颈鳞癌组Ile/Val、Val/Val基因型的分布频率明显高于对照组。Ile/Val和Val/Val基因型的个体发生宫颈鳞癌的OR值分别是Ile/Ile基因型个体的1.969倍和3.15倍,差异有统计学意义(P<0.05);(2)CYP1A1MspⅠ位点多态性分析:将MspⅠ位点的PCR产物行酶切分析,显示m1/m2杂合型、m2/m2突变型在宫颈鳞癌组和对照组表达差异无统计学意义(P>0.05)。结论:宫颈鳞癌组织中CYP1A1基因Exon7位点突变率升高,Ile/Val和Val/Val基因突变型个体发生宫颈鳞癌的危险性明显升高;CYP1A1基因MspⅠ位点的多态性可能与宫颈鳞癌易感性无关。     

两个Alstrom综合征家系 ALMS1基因变异分析及产前诊断

本研究报道了2个Alstrom综合征家系的遗传学病因, 并据此对再次妊娠的胎儿进行产前诊断。这2个家系先证者均存在不同程度视力异常, 现先证者母亲均再次妊娠, 就诊于郑州大学第一附属医院进行产前诊断。全外显子组测序及Sanger测序验证结果显示, 家系1的先证者为ALMS1基因c.6103C>T(p.Gln2035*)和c.6430C>T(p.Arg2144*)复合杂合变异, 父母分别为携带者;家系2的先证者为ALMS1基因c.9148₉149delCT(p.Cys3051Serfs*9)和c.12028delC(p.Leu4010Typfs*19)复合杂合变异, 父母分别为携带者;其中c.6103C>T(p.Gln2035*)、c.9148₉149delCT(p.Cys3051Serfs*9)和c.12028delC(p.Leu4010Typfs*19)均为新发现的变异位点。家系1胎儿携带c.6430C>T(p.Arg2144*)杂合变异, 遗传咨询后选择继续妊娠;家系2胎儿与先证者相同, 携带c.9148_...     

应用变性高效液相色谱技术检测结节性硬化症TSC1基因外显子15基因突变的研究

摘要：目的　探讨结节性硬化症（TSC）TSC1基因外显子15基因突变的特点。方法　1996—2006年采用多聚酶链扩增和变性高效液相色谱（DHPLC）技术,对21个家系59名研究对象进行了TSC1基因外显子15检测,对DHPLC检测异常者再进行多聚酶链扩增产物直接测序方法证实其突变类型。结果　对21个家系TSC1 基因外显子15共检测出4个家系的3种突变形式,其中c.1708～1709delAG（p.Arg570GlyfsX17）与c.1888～1891delAAAG（p.Lys630GlnfsX22） 两种突变为国内尚未报道的小的缺失突变,c.1460C > G（p.Ser487Cys）突变为1种罕见的错义突变。TSC1 基因外显子15突变基因检出频率为4/21（19%）。在检出突变的4个家系中1个为家系突变,其余3个为散发突变。结论　TSC1基因外显子15的c.1460C > G（p.Ser487Cys）,c.1708～1709delAG（p.Arg570GlyfsX17）与c.1888～1891delAAAG （p.Lys630GlnfsX22） 突变为目前国内首报突变。     

CYP1A1基因多态性与子宫内膜腺癌发生的关系研究

目的研究细胞色素P4501A1(CYP1A1)基因MspI位点和Ile-Val位点多态性与子宫内膜腺癌发生的关系。方法以病例-对照研究,采用聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)方法和等位基因特异性PCR(AS-PCR)技术检测84例子宫内膜腺癌患者和66例对照者CYP1A1基因MspI位点和Ile-Val位点的多态性。结果Ile-Val三种多态基因型在对照组和子宫内膜腺癌组中的分布在显著性差异(P<0.05),其中Ile/Val和Val/Val基因型在子宫内膜腺癌组中的分布频率明显高于对照组。与Ile/Ile基因型相比,Ile/Val基因型个体发生子宫内膜腺癌的相对危险度(OR)是2.682,两者差异有统计学意义(P<0.05);而MspI位点的多态性在对照组和子宫内膜腺癌组中的分布差异无统计学意义(P>0.05)。结论CYP1A1基因第7外显子的Ile/Val基因型与子宫内膜腺癌的发生有关,可以作为子宫内膜腺癌易感人群筛查的重要指标,MspI位点多态性与子宫内膜腺癌的发生无相关性。     

葡萄糖醛酸转移酶1A1 G71R突变和葡萄糖 6 磷酸脱氢酶缺陷对新生儿胆红素浓度的影响

目的探讨葡萄糖醛酸转移酶1A1(UGT1A1)基因G71R突变、葡萄糖 6 磷酸脱氢酶(G6PD)缺陷对新生儿生后前3d胆红素浓度的影响。 方法测定81例新生儿脐血的G6PD活性及G71R基因型，分组比较生后前3d光疗前胆红素值的组间差异。用等位基因特异性寡核苷酸探针点杂交法(ASO)确定G71R基因型。 结果在G71R野生型新生儿中，G6PD缺乏组与G6PD正常组相比，生后前3d胆红素值间无统计学差异。G6PD正常新生儿中,G71R突变纯合子或杂合子的新生儿生后前3d胆红素浓度与G71R正常野生型新生儿相比无统计学差异。G6PD缺陷新生儿中，同时合并有G71R突变纯合子或杂合子的新生儿组生后第2天、第3天胆红素浓度高于G71R正常野生型新生儿组。 结论G6PD缺乏与G71R基因突变并存加重新生儿黄疸程度。     

HADHB基因变异导致致死型线粒体三功能蛋白缺乏症1例

本文报告了1例HADHB基因复合杂合变异导致的致死型线粒体三功能蛋白缺乏症(mitochondrial trifunctional protein deficiency, MTPD)。患儿生后24 h内起病, 以"吃奶少伴心肌酶异常"为主诉在生后第4天转至南京医科大学附属儿童医院。入院时体格检查见精神稍萎靡, 反应尚可, 心肺腹无明显异常;实验室检查肌酸激酶同工酶及肌钙蛋白均偏高, 心电图提示窦性心动过速、肢导联低电压;血液代谢筛查提示十四烯酰基肉碱及多种3-羟基肉碱均偏高;全外显子组测序结果示胎儿HADHB基因复合杂合变异[c.739C>T(p.Arg247Cys)和c.607C>T(p.Arg203Ter, 272)]。随访至3月龄, 患儿受凉后出现吃奶差、精神差, 再入院后出现心力衰竭、呼吸衰竭等多脏器功能障碍表现, 经治疗无效后死亡。致死型MTPD较为罕见, 目前无有效治疗措施, 预后差。对于新生儿期出现不明原因心肌病、低血糖、酸中毒等代谢异常表现, 应警惕致死型MTPD可能, 新生儿期基因检测可早期明确诊断。     

哈萨克族肥胖儿童β3-肾上腺素能受体基因多态性观察

目的探讨β3-肾上腺素能受体基因(β3-AR)Trp64Arg变异与新疆哈萨克族儿童肥胖的相关性。方法选取乌鲁木齐周边地区95例6~12岁哈萨克族学龄肥胖儿童及87名非肥胖儿童，用限制性片段长度多态性方法检测被调查儿童的基因型，生化方法检测血清甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、载脂蛋白Ａ(ApoA)、载脂蛋白Ｂ(ApoB)水平，并测量身高、体重。结果变异型等位基因(Ｃ)在被调查对象中出现的频率为0.194，其中男0.210，女0.160。肥胖儿童中变异等位基因(Ｃ)、Trp64Arｇ变异基因型的出现频率明显高于非肥胖者(P<0.05)。单纯性肥胖儿童与非肥胖儿童比较，血清TG、TC、LDL-C、ApoB水平均明显升高(P<0.05)。β3-AR不同基因型间血脂比较，差异无统计学意义(P>0.05)。结论哈萨克族学龄儿童中存在一定的β3-AR Trp64Arg变异，可能与哈萨克族儿童肥胖有关。     

应用突变引物延伸扩增技术快速产前诊断β地中海贫血基因突变

目的 探讨综合应用突变引物延伸扩增(MOEA)及巢式聚合酶链反应(PCR)技术进行β地中海贫血产前诊断的可行性.方法 先用一对外侧引物扩增1.5 Kb β珠蛋白基因片段,再用MOEA技术检测β地中海贫血点突变.结果 完成了6例β地中海贫血高危胎儿的产前基因诊断,其中双重杂合子1例,杂合子3例,正常胎儿2例;经分娩或流产后基因分析验证,结果与产前诊断完全一致.结论 MOEA技术适合于已知点突变的基因诊断和产前诊断,具有快速、简便、安全、准确等优点,便于推广应用.     

高通量测序技术鉴定新生儿窒息性胸廓发育不良症1例

目的对1例多器官发育缺陷的新生儿进行高通量全外显子组测序(WES),明确遗传学病因。方法对患儿进行临床检查,采集患儿及其父母外周静脉血,进行全外显子组基因测序与分析,再对可疑突变位点进行Sanger测序验证。结果患儿临床表现为胸廓发育畸形伴先天性心脏病和肝脾肿大,全外显子组测序显示其存在DYNC2H1基因c.8512CT(p.R2838*)及c.10163CT(p.P3388L)复合杂合突变,分别遗传自父亲和母亲。结论根据测序结果结合临床表现,鉴定该患儿为DYNC2H1基因复合杂合突变引起的窒息性胸廓发育不良(ATD)。     

以转氨酶升高为主的多种酰基辅酶A脱氢酶缺乏症1例临床表现及基因分析

目的研究1例以转氨酶升高为主的多种酰基辅酶A脱氢酶缺乏症患者临床表现、实验室检查、肌肉活检及基因突变情况分析,并进行文献复习,为该病的早期诊断及治疗提供依据。方法收集1例7岁3月男性患儿的临床资料,采集患儿及父母血标本,采用二代基因测序检测致病基因,腓肠肌穿刺活检明确肌肉病变情况。结果患儿肌肉活检电镜结果示肌纤维内大量脂滴沉积。基因测序结果显示患儿的ETFDH基因存在c.1773_1774del AT p.(Cys592※)无义突变和c.389AT p.(Asp130Val)错义突变,考虑为复合杂合突变,患儿父母分别为携带者。结论临床上有转氨酶升高为主伴有心肌酶升高、运动障碍者,应尽早进行分子遗传学检查,有条件者行腓肠肌肌肉活检术,可以为患儿家庭提供准确的遗传咨询和产前诊断。     

Crigler-Najjar syndrome type 2.

Crigler-Najjar syndrome is a rare disorder of bilirubin metabolism with two distinct forms: type 1 and type 2. We report three patients with Crigler-Najjar syndrome type 2 (CN-2). All patients had serum bilirubin values higher than 171 micromol/L and deep yellow skin color. The results of other liver function tests, glucose-6-phosphate dehydrogenase activity and hematology tests were normal, and immunologic tests for hepatitis A, B and C were negative, although one patient had slightly elevated alanine aminotransferase level (45 IU/L). Polymerase chain reaction and sequence analysis of the UDP-glucuronosyltransferase 1A1 (UGT1A1) gene revealed a novel homozygous T>A mutation at nucleotide 479 in exon 1 (Val160Glu) of patient 1, a novel homozygous A>G mutation at nucleotide 610 in exon 1 (Met204Val) of patient 2, and a homozygous T>G variation at nucleotide 1456 in exon 5 (Tyr486Asp) plus a heterozygous G>A variation at nucleotide 211 in exon 1 (Gly71Arg/normal) of patient 3. Two of these mutations were novel and variations identified within the coding region of the UGT1A1 gene were considered the cause of CN-2 in all three patients.     

FSH β-subunit mutations in two sisters: the first report from the Indian sub-continent and review of previous cases

Isolated FSH deficiency due to mutations in the gene for β-subunit of FSH is an extremely rare autosomal recessive disease of which only eleven cases have been reported so far. The clinical features include absent breast development and primary amenorrhea in females and azoospermia with normal testosterone levels in males. In this study we report two Kashmiri sisters born to native Kashmiri consanguineous parents with failure of onset of puberty. Hormonal evaluation revealed undetectable serum FSH and estradiol and high LH. Genetic analysis of FSH β-gene identified one nonsense mutation (c.343C?>?T:p. Arg115Stop) in exon 3. The two sisters were homozygous for this nonsense mutation while the parents were heterozygous. Incorporation of a stop codon at 115 codon position is predicted to result in the formation of truncated FSH β protein, lacking 14 amino acid from the carboxy-terminus (p.Arg115Stop). Very recently, this same mutation was reported for the first time in a Chinese male. Ours is the first ever report of any FSH β-subunit mutation from the Indian sub-continent and this particular mutation in any female from anywhere in the world. We conclude and emphasize that this diagnosis should be considered in girls with delayed puberty and selective deficiency of FSH.     

Small mutations of the DMD gene in Taiwanese families

BACKGROUND/PURPOSE: Duchenne/Becker muscular dystrophies are X-linked recessive disorders caused by mutations in the Duchenne muscular dystrophy (DMD) gene. We aimed to demonstrate the small mutation patterns of the DMD gene in Taiwanese subjects. METHODS: We sequenced all 79 exons of the DMD gene in 33 unrelated Taiwanese families in which large-scale deletions and duplications had been excluded by multiplex ligation-dependent probe amplification. RESULTS: Direct sequencing detected 23 different mutations from 26 families, including 15 novel mutations and eight previously reported ones. The 15 novel mutations consisted of seven substitutions (c.1238C>G [p.S413X], c.2971G>T [p.E991X], c.3172C>T [p.Q1058X], c.7402G>T [p.E2468X], c.8022C>G [p.S2605X], c.10018T>C [p.C3340R], c.10546G>T [p.E3516X]), six small deletions (c.2202delG [p.A668fsX676], c.2268delC [p.F756fsX759], c.4611delT [p.N1537fsX1545], c.4856_4857delAA [p.K1619fsX1621], c.6638delT [p.L2213fsX2220], c.9457delT [p.C3153fsX3154]), and two small insertions (c.4351insA [p.L1451fsX1468], c.10493_10495insAAT [p.L3498X]). Twenty-two of the 23 pathologic changes disrupted the translational reading frame (13 nonsense, 7 frameshift, 2 splice-site change), whereas only one was a missense variant with known pathogenic nature. Two previously reported mutations, c.8038C>T [p.R2680X] and c.10108C>T [p.R3370X] were detected in two and three unrelated families, respectively. CONCLUSION: Most identified mutations either led to a predictable premature stop codon or resulted in splicing defects, which caused defective function of dystrophin. Our findings extend the mutation spectrum of the DMD gene. Molecular characterization of the affected families is important for genetic counseling and prenatal diagnosis.     

KLHL40基因复合杂合变异导致先天性线状体肌病一例

本文报道了1例KLHL40基因复合杂合突变导致线状体肌病的病例。患儿女,生后即起病,以"气促、肌力、肌张力低"为主要表现,常规治疗后症状加重,体格检查示四肢肌力低、肌张力低,原始反射均未引出;血清肌酸激酶升高。患儿未行肌肉活检。采用全外显子组二代测序证实患儿存在KLHL40基因复合杂合突变,包括c.932G>T(p.R311L)和c.1487T>A(p.M496K),并且分别来源于父母,其中c.932G>T(p.R311L)为已发现的突变位点,c.1487T>A(p.M496K)为新发现的突变位点。线状体肌病是一种罕见的先天性肌肉疾病,肌纤维中发现线状体是特征性肌肉病理改变。病理及基因诊断是诊断线状体肌病的金标准。     

Metaphyseal chondrodysplasia McKusick type in a Chinese fetus, caused by novel compound heterozygosity 64T> A and 79G >T in RMRPgene

We present the first confirmed case by molecular analysis of a metaphyseal chondrodysplasia, McKusick type, in a 22-week fetus. Two novel compound heterozygous mutations, 64T> A and 79G > T, were found in the highly conserved regions of the RMRP gene. Twenty-two heterozygous g.1018 T> C mutations, two homozygous g.1018 T> C mutations, two heterozygous insertion mutations g.799_g.800insC and one heterozygous insertion mutation g.849_g.850insT were found among 100 normal controls. Careful radiological examination of the fetus for skeletal dysplasia allowed definitive diagnosis, proper genetic counselling and future prenatal diagnosis.     

Mucopolysaccharidosis type VI: Report of two Taiwanese patients and identification of one novel mutation.

Mucopolysaccharidosis type VI (MPS VI) is an autosomal recessive lysosomal storage disease caused by a deficiency of N-acetylgalactosamine-4-sulphatase (arylsulfatase B, ASB). We report the clinical investigation and mutation analysis of two Taiwanese patients with severe (Case 1) and intermediate (Case 2) phenotypes of MPS VI. Three missense mutations and one polymorphism were identified. Case 1 was found to have a novel heteroallelic C-to-G transversion at nucleotide 1197 causing a phenylalanine to leucine substitution at residue 399 (Phe399Leu), and a heteroallelic Gln239Arg mutation. In Case 2, a heterozygous Cys192Arg mutation and a Val358Met polymorphism were identified. Among these three mutations, the Gln239Arg and Phe399Leu substitutions have so far been observed only in the Taiwanese population. The correlation between genotype and phenotype contributes to molecular pre- and post-natal diagnosis for MPS VI patients.     

Analyses of GDF9 mutation in 100 Chinese women with premature ovarian failure

We screened growth differentiation factor 9 coding regions for mutations in a Chinese sample of 100 women with premature ovarian failure and discovered four novel single-nucleotide polymorphisms: c.436C>T (p.Arg146Cys), c.588A>C (silent), c.712A>G (p.Thr238Ala), and c.1283G>C (p.Ser428Thr). Nonsynonymous single-nucleotide polymorphisms c.436C>T and c.1283G>C were also detected in the control population. The c.712A>G perturbation results in a missense mutation (p.Thr238Ala) and was not present in any of 96 controls. Substitution of the hydrophobic amino acid residue alanine for hydrophilic threonine may disrupt growth differentiation factor 9 function.     

全外显子测序检测6个囊性肾病胎儿及其核心家系的致病突变

目的全外显子测序技术检测6个囊性肾病胎儿及其核心家系,寻找致病变异,为遗传咨询、产前诊断或植入前诊断提供遗传学数据支持。方法应用Illumina Hiseq2500测序平台,对超声检查提示为囊性肾病、染色体微阵列检测(CMA)未见异常的胎儿标本进行家系全外显子检测,参考数据库Human Genome 19(hg19/GRCh37),根据美国医学遗传学与基因组学学会指南(ACMG,2015)及指南应用建议评估变异致病性,对评级可疑致病位点应用Sanger测序验证。结果家系WES检出3个家系存在异常突变,其中1个家系的突变为遗传性,2个家系胎儿的突变为新发突变;余3个家系未检出明确致病突变。胎儿1 PKHD1基因c.5869G>A(父源)和c.9455delA复合杂合突变(母源),均为可能致病的变异。胎儿5 HNF1B基因c.826C>T杂合突变,为致病性新发突变。胎儿6 HNF1B基因c.1318G>T杂合突变,为可能致病性新发突变。Sanger测序验证结果与全外显子检测结果一致。结论对囊性肾病可运用家系核心成员全外显子检测,筛选出候选变异后进行Sanger测序验证,并对家系进行遗传分析,有助于遗传咨询、再次妊娠的产前诊断或辅助生殖的植入前诊断。本研究检出3个未见文献报道的可能致病突变位点,1个致病性突变位点已在个别文献报道但是未在国际公认数据库明确为致病突变位点,本研究拓宽了囊性肾病的致病基因位点谱。     

NovelCFAP43 andCFAP44 mutations cause male infertility with multiple morphological abnormalities of the sperm flagella (MMAF)

Research question

Multiple morphological abnormalities of the sperm flagella (MMAF) comprise a rare congenital disease that can cause primary male infertility. Several pathogenic genes (e.g. AKAP4, DNAH1, CFAP43 and CFAP44) are associated with MMAF but the pathogenic mechanisms have not been elucidated.