一个常染色体显性遗传Emery-Dreifuss型肌营养不良家系的基因突变分析

Objective To investigate the clinical, pathological and genetic characteristics in a family with autosomal dominant Emery-Dreifuss muscular dystrophy (AD-EDMD). Methods Clinical data and skeletal muscle specimens were collected from two patients (the proband and her daughter) for pathological analysis. DNA samples of the proband and her family members (7 persons from 3 generations) were obtained for PCR amplification and direct DNA sequencing of the lamin A/C (LMNA) gene. Haplotype analysis was performed after the identification of mutation. Results The proband had typical clinical manifestation of EDMD: joint contracture, progressive muscle weakness and atrophy and cardiac conduction dysfunction. Muscular pathology revealed myopathic changes combined with slight neuropathic changes. A heterozygous missense mutation 1583 (C→G) (T528R) was identified in exon 9 of the LMNA gene in the two patients, but not in other family members. Haplotype analysis indicated that the proband and her daughter shared the same causative haplotype. Conclusion This is the first report of the phenotype and genotype of AD-EDMD in Chinese.     

一个常染色体显性遗传Emery-Dreifuss型肌营养不良家系的基因突变分析

Objective To investigate the clinical, pathological and genetic characteristics in a family with autosomal dominant Emery-Dreifuss muscular dystrophy (AD-EDMD). Methods Clinical data and skeletal muscle specimens were collected from two patients (the proband and her daughter) for pathological analysis. DNA samples of the proband and her family members (7 persons from 3 generations) were obtained for PCR amplification and direct DNA sequencing of the lamin A/C (LMNA) gene. Haplotype analysis was performed after the identification of mutation. Results The proband had typical clinical manifestation of EDMD: joint contracture, progressive muscle weakness and atrophy and cardiac conduction dysfunction. Muscular pathology revealed myopathic changes combined with slight neuropathic changes. A heterozygous missense mutation 1583 (C→G) (T528R) was identified in exon 9 of the LMNA gene in the two patients, but not in other family members. Haplotype analysis indicated that the proband and her daughter shared the same causative haplotype. Conclusion This is the first report of the phenotype and genotype of AD-EDMD in Chinese.     

Liddle's syndrome caused by a novel mutation of the γ-subunit of epithelial sodium channel gene SCNN1G in Chinese

Objective To screen the mutation of the β and γ subunits of epithelial sodium channel gene SCNN1 in two families with Liddle's syndrome. Methods Two patients clinically diagnosed as Liddle's syndrome and their family members were enrolled. Peripheral blood samples were collected and total genomic DNA was prepared. Polymerase chain reaction (PCR) was used to amplify the exon 13 of the SCNN1B and SCNN1G gene. PCR products were purified and subjected to direct DNA sequencing. Results A heterozygous nonsense mutation at codon 564 of the SCNN1B gene from CGA(Arg) to stop codon(TGA)was detector in the proband of family 1. More importantly, a novel heterozygous nonsense mutation of CAG (Gln) to stop codon TAG at codon 567 of the SCNN1G gene was detected in the proband and another two members of family 2. Conclusion Screening for specific mutations of the SCNN1 gene in relatives of patients with Liddle's syndrome can be used to identify the previously unrecognized cases within the family.A new nonsense mutation(Q567X) of the SCNN1G gene is likely the cause of Liddle's syndrome in family 2.     

Liddle's syndrome caused by a novel mutation of the γ-subunit of epithelial sodium channel gene SCNN1G in Chinese

Objective To screen the mutation of the β and γ subunits of epithelial sodium channel gene SCNN1 in two families with Liddle's syndrome. Methods Two patients clinically diagnosed as Liddle's syndrome and their family members were enrolled. Peripheral blood samples were collected and total genomic DNA was prepared. Polymerase chain reaction (PCR) was used to amplify the exon 13 of the SCNN1B and SCNN1G gene. PCR products were purified and subjected to direct DNA sequencing. Results A heterozygous nonsense mutation at codon 564 of the SCNN1B gene from CGA(Arg) to stop codon(TGA)was detector in the proband of family 1. More importantly, a novel heterozygous nonsense mutation of CAG (Gln) to stop codon TAG at codon 567 of the SCNN1G gene was detected in the proband and another two members of family 2. Conclusion Screening for specific mutations of the SCNN1 gene in relatives of patients with Liddle's syndrome can be used to identify the previously unrecognized cases within the family.A new nonsense mutation(Q567X) of the SCNN1G gene is likely the cause of Liddle's syndrome in family 2.     

建立一种新型凝血因子Ⅴ基因突变小鼠模型：突变小鼠无显著骨组织变化

BACKGROUND: Blood coagulation factor V gene mutation in non-traumatic femoral head necrosis has been shown to have an higher incidence than that in healthy and secondary non-traumatic femoral head necrosis, and the incidence of thrombosis is positively related. Inactivated blood coagulation factor V can accelerate the activation of prothrombin and the generation of thrombin. Mutations at arg-306, arg-506 and arg-679 will result in the blood clots and hypercoagulable state. Here, this study is designed to investigate the influence of R506Q/R679Q on osteonecrosis. OBJECTIVE: To establish the mouse model of mutations of Gln506Arg and Gln679Arg in coagulation Factor V (Factor VR506Q/R679Q). METHODS: Factor VR506Q/R679Q point mutation target vector was constructed by molecular cloning technology, the linearization vector was transfected into embryonic stem cells, and then G418-resistant cells were screened and used for microinjection. The target blastocysts were transplanted to the fallopian tube of estrus mice to obtain the Chimera mice carrying bilateral LoxP gene, followed by mated with CMV-cre transgenic mice, and then the mice with Factor VR506Q/R679Q point mutations were obtained. After genotype identification by PCR, hematoxylin-eosin staining results and percentage of empty lacunae were compared between the mutant and wild-type mice, and rat bone tissue and bone mass were analyzed. RESULTS AND CONCLUSION: There were no obvious abnormalities in the embryonic and postnatal development, percentage of empty lacunae and bone mass of Factor VR506Q/R679Q point mutation mice when compared with the wild-type mice. These results suggest that the mouse model with Factor VR506Q/R679Q point mutation is established successfully, but there is no significant change in the bone tissue. The following research should focus on the effect of external stimulus on the incidence of osteonecrosis in a mutant mouse. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Analysis of clinical phenotype and ACAT1 gene mutation in a family affected with β-ketothiolase deficiency北大核心CSCD

Objective: To investigate the clinical phenotype and ACATI gene mutation in a family affected with β-ketothiolase deficiency (BKTD). Methods: Clinical features and laboratory test data were collected. The probands were monozygotic twin brothers. Genomic DNA was isolated from peripheral blood leukocytes obtained from the probands and their family members. Molecular genetic testing of the ACAT1 gene was carried out. Results: The probands have presented with fever, vomiting and severe ketoacidosis. By arterial blood gas testing, pH was determined to be 7. 164, bicarbonate was 4. 0 mmol/L, and urine ketone was + + + +. Urinary organic acid gas chromatography-mass spectrometry analysis showed excessive excretion of 3-hydroxybutyric acid, 2-methyl-3-hydroxybutyric acid and tiglylglycine. Increased 3-hydroxybutyrylcarnitine (C4-OH), tiglylcarnitine (C5:1) and 3-hydroxyisovalerylcarnitine (C5-0H) levels. The clinical phenotype of proband's parents were both normal, but an elder sister turned out to be an affected patient. Genetic analysis has identified two heterozygous mutations [c. 622C>T(p. R208X) and c. 653C>T(p. S218F)] in the proband, which were respectively detected in the mother and father. The c. 653C>T(p. S218F) mutation was not found among the 100 healthy controls and has not been included in the Human Gene Mutation Database (HGMD). Conclusion: The primary clinical manifestations of BKTD is ketoacidosis. Urine organic acid and blood acylcarnitine analyses play an important role in the diagnosis of the disease. The compound heterozygous of ACAT1 gene mutations probably underlie the BKTD in our patient. © 2016, West China University of Medical Sciences. All rights reserved.     

中国人黏多糖贮积症ⅣA型家系GALNS基因新突变的鉴定

目的 研究黏多糖贮积症ⅣA型(mucopolysaccharidosis type ⅣA,MPS ⅣA)患者发病的分子遗传学机制,揭示其基因型与表现型的相互关系,为产前基因诊断等创造必要的前提条件.方法 采用尿糖胺聚糖(glycosaminoglycans,GAGs)定性检测法对疑似MPS ⅣA型的先证者进行初诊,然后采用PCR及扩增产物直接测序法对先证者及其家庭成员进行突变检测.在检出GALNS基因c.1567T＞G新突变后,先后建立XspⅠ酶切鉴定法和扩增阻碍突变系统(amplification refractory mutation system,ARMS)快速特异鉴定法,对随机采集的110名正常对照与先证者及其家庭成员的GALNS基因第14外显子进行序列分析,同时采用生物信息学方法对蛋白质二级、三级结构进行预测,以及直接测定患儿GALNS酶活性的方法,对该新突变进行致病性鉴定.结果 先证者尿检呈弱阳性GAGs(±),其GALNS基因第14外显子内存在杂合的c.1567T＞G终止密码突变,第4外显子存在杂合的c.374C＞T错义突变,为两种突变的复合杂合子.其妹突变类型与先证者完全相同,其母仅在第14外显子存在杂合的终止密码突变,为该病的携带者,其父仅在第4外显子存在杂合的错义突变,也为杂合子;第14外显子的PCR产物经XspⅠ酶切后,正常对照组切出28 bp、120 bp、399 bp 3条带,而患者和携带者的母亲均切出28 bp、120 bp、148 bp和399 bp 4条带;用ARMS特异引物扩增后,正常对照组均扩增阴性,而患者及携带者均扩增阳性;蛋白质二级、三级结构预测结果显示:c.1567T＞G变异导致终止密码(TAG)突变为谷氨酸(GAG),使多肽链延长了92个氨基酸残基,导致蛋白质二、三级结构发生明显改变,而正常对照无此变化.酶活性测定的结果显示:患者的GALNS酶活性仅为8.3 nmol/17 h/mg pr,明显低于正常值(正常参考值为41.9～92.1 nmol/17h/mg pr).结论 c.1567T＞G变异是一种新的致病性突变,是引起该家系患儿发病的根本原因.

Abstract：

Objective To study the molecular genetic mechanism of mucopolysaccharidosis type ⅣA(MPS ⅣA), and reveal the relationship between the genotype and phenotype, and provide a basis for prenatal gene diagnosis in the future. Methods A preliminary diagnosis was made by qualitative detection of urinary glycosaminoglycans of the suspected MPS ⅣA proband. Then, mutation detection was performed on the proband and her family members with PCR and direct sequencing of the PCR products. After a novel c.1567T＞G mutation was detected,XspⅠrestriction enzyme digestion and amplification refractory mutation system(ARMS) fast specific identification were established to analyze the sequences of exon 14 in GALNS gene, including 110 randomly selected healthy controls, the proband and other pedigree members. At the same time, bioinformatic approaches for protein secondary, tertiary structure prediction were applied to identify the novel pathologic mutation. Results The proband's urine GAGs test was a weak positive(±),and a c.1567T＞G heterozygous termination codon mutation in exon 14 and a c.374C＞T heterozygous missense mutation in exon 4 were found. The proband was compound heterozygous of the two mutations, so was her younger sister. Her mother was a carrier with only a c.1567T＞G heterozygous mutation in exon 14. Her father had a heterozygous mutation of c.374C>T in exon 4. After XspⅠrestriction enzyme digestion, healthy controls had three bands including 28 bp, 120 bp and 399 bp, while the proband and her mother had four bands consisting of 28 bp, 120 bp,148 bp and 399 bp. For amplification by ARMS specific primers, it was negative for the controls,while it was positive for the proband and the carrier. The results of protein secondary and tertiary structure prediction showed that the c.1567T＞G mutation located in the stop codon, resulted in stop codon (TAG) changing to glutamic acid (GAG), with the peptide chain extending 92 amino acid residues, and secondary and tertiary protein structure change, which were not found in the controls. The result of enzyme assay showed that the activity of GALNS enzyme in the affected child was 8.3 nmol/17h/mg pr, which was obviously lower than the normal value (the normal range is 41.9-92.1 nmol/17h/mg pr). Conclusion These results illustrate that the c.1567 T＞G is a novel pathologic mutation, which is the main cause of the disease in this family.     

两个新RUNX2基因突变引起家族性锁骨颅骨发育不全

Objective To identify the RUNX2 gene mutation in two unrelated Chinese families with cleidocranial dysplasia (CCD), and to assess the feasibility of gene diagnosis for patients with CCD. Methods Genomic DNA was isolated from peripheral blood samples of 4 patients and 4 healthy members in the two pedigrees as well as 102 unrelated healthy controls. All 7 coding exons and their flanking intronic sequences of the RUNX2 gene were amplified by PCR, then the PCR products were sequenced bi-directionally. The sequencing results were compared with normal sequences in GenBank to identify the mutation. The mutation was confirmed by RFLP with restriction endonuclease. Results In one family, a novel heterozygous missense mutation c. 346T＞A (W116R) in exon 1 of the RUNX2 gene was detected in the two affected individuals, and the mutation was further confirmed with Bsr Ⅰ restriction endonuclease digestion. In the other family, a novel nonsense mutation c. 610A＞T (K204X) was identified in the two patients. No above sequence change was found in the 102 healthy controls. Conclusion Two novel RUNX2 mutations were found in two unrelated Chinese families with cleidoeranial dysplasia. The identification of these mutations further extended the mutation spectrum of RUNX2 gene and will facilitate prenatal diagnosis and gene diagnosis of CCD.     

两个新RUNX2基因突变引起家族性锁骨颅骨发育不全

Objective To identify the RUNX2 gene mutation in two unrelated Chinese families with cleidocranial dysplasia (CCD), and to assess the feasibility of gene diagnosis for patients with CCD. Methods Genomic DNA was isolated from peripheral blood samples of 4 patients and 4 healthy members in the two pedigrees as well as 102 unrelated healthy controls. All 7 coding exons and their flanking intronic sequences of the RUNX2 gene were amplified by PCR, then the PCR products were sequenced bi-directionally. The sequencing results were compared with normal sequences in GenBank to identify the mutation. The mutation was confirmed by RFLP with restriction endonuclease. Results In one family, a novel heterozygous missense mutation c. 346T＞A (W116R) in exon 1 of the RUNX2 gene was detected in the two affected individuals, and the mutation was further confirmed with Bsr Ⅰ restriction endonuclease digestion. In the other family, a novel nonsense mutation c. 610A＞T (K204X) was identified in the two patients. No above sequence change was found in the 102 healthy controls. Conclusion Two novel RUNX2 mutations were found in two unrelated Chinese families with cleidoeranial dysplasia. The identification of these mutations further extended the mutation spectrum of RUNX2 gene and will facilitate prenatal diagnosis and gene diagnosis of CCD.     

Detection of mutation in embB gene of Mycobacterium tuberculosis from clinical isolates of tuberculous patients in China by means of reverse-dot blot hybridization

The relationship between embB mutation of Mycobacterium tuberculosis and ethambutol (EMB) resistance of the clinical isolates of tuberculous patients in China was investigated by reverse dot blot hybridization (RDBH) in addition to evaluating the clinical value with application of PCR-RDBH technique to detect EMB resistance. In the present study, the genotypes of the 258 bp fragments of embB genes from 196 clinical isolates of M. tuberculosis were analysed with RDBH and DNA sequencing. It was demonstrated that 60 out of 91 phenotypically EMB-resistant isolates (65.9%) showed 5 types of missense mutations at codon 306 of embB gene, resulting in the replacement of the Met residue of the wild type strain with Val, Ile or Leu residues. In these mutations, the GTP mutation (38/91, 41.8%) and the ATA mutation (16/91, 17.6%) were the most encountered genotypes. The embB mutation at codon 306 could also be found in 69 isolates of phenotypically EMB-sensitive but resistant to other anti-tuberculous drugs, but no such gene mutation could be found in 36 strains of drug-sensitive isolates. Meanwhile, the concordance with the results of DNA sequencing for one wide-type probe and 5 probes for specific mutations was 100% . It was concluded that the EMB-resistance occurring in most M. tuberculosis is due to appearance of embB mutation at codon 306, and the PCR-RDBH assay was proved to be a rapid, simple and reliable method for the detection of gene mutations, which might be a good alternative for the drug-resistance screening.     

一个糖原累积病Ⅱ型家系的酸性-α-葡萄糖苷酶及其产前基因诊断

目的 为1个糖原累积病Ⅱ型(glycogen storage disease typeⅡ,GSDⅡ)家系进行酶学和产前基因诊断.方法 用酸性-α-葡萄糖苷酶(acid-alpha-glucosidase,GAA)特异性水解荧光底物4-甲基伞型酮-α-D-吡喃葡萄糖苷(4-methylumbelliferyl-α-D-glucopyranoside,4-MUG)和阿卡波糖抑制其同工酶的方法检测外周血白细胞和羊水细胞GAA酶活性,聚合酶链反应扩增GAA基因外显子编码区序列,直接测序分析GAA基因突变情况.结果 先证者外周血白细胞与胎儿羊水细胞GAA酶活性均明显低于正常参考值范围,分别为正常对照平均值的12.3% 和1.1%.先证者和胎儿均携带新无义突变 p.W738X 和已报道的无义突变p.E888X;先证者、母亲和胎儿均携带假性缺陷等位基因[c.1726G>A; c.2065G>A].结论 通过GAA酶活性检测结合GAA基因分析对1个GSDⅡ家系进行了产前诊断.由于假性缺陷等位基因可引起GAA酶活性降低,故GAA基因分析应作为亚洲人群GSDⅡ产前诊断的常规手段.

Abstract：

Objective To carry out prenatal diagnosis for a glycogen storage disease typeⅡ(GSDⅡ) affected family. Methods The acid-α-glucosidase (GAA) activity was measured in whole leukocytes and cultured amniocytes with 4-methylumbelliferyl-α-D-glucopyranoside as substrate and with acarbose as inhibitor. The coding regions of GAA gene were amplified by polymerase chain reaction and analyzed by direct DNA sequencing. Results The proband and the fetus had low GAA activity (12.3% and 1.1% of the average normal range, respectively). Mutation analysis of the GAA gene revealed a novel nonsense mutation p.W738X and a reported nonsense mutation p.E888X in both the proband and the fetus; the reported pseudodeficiency allele c.[1726G>A;2065G>A] was found in the proband, the mother and the fetus. Conclusion The proband and the fetus were both GSDⅡaffected. A combination of GAA activity analysis and mutation analysis is efficient for the prenatal diagnosis of GSDⅡ. Mutation analysis should be a routine method in the prenatal diagnosis of GSDⅡ in Asian population, where pseudodeficiency allele can cause low GAA activity in normal individuals which is relatively common in Asian.     

九例遗传性凝血因子Ⅶ缺陷症患者的分子发病机制与临床特性

目的 探讨9个遗传性凝血因子Ⅶ( coagulation factorⅦ,FⅦ)缺陷症家系的基因突变类型与临床特征.方法 检测凝血酶原时间、活化部分凝血活酶时间及FⅦ活性和抗原等指标以明确诊断;用PCR法扩增先证者F7基因的全部外显子及侧翼序列、5′和3′非翻译区.PCR产物纯化后直接测序,寻找基因突变,用反向测序证实所发生的突变.结果 9个家系中的先证者凝血酶原时间均延长,FⅦ活性在2.0％～6.0％之间,7例先证者的FⅦ抗原显著减低.共发现8种F7基因突变,其中g.8355 A＞T(Gln100Leu)、g.11243T＞C(Ser269Pro)和g.11520_11521insT 3种突变为首次发现;6种突变发生在催化区;缺失突变和插入突变各1种,其余均为错义突变;所有的基因突变均来自先证者的父亲和(或)母亲,发现5个家系存在近亲婚配.g.27_28delCT、Cys329Gly、Arg304Trp和His348Gln突变在无亲缘关系的家系中重复出现.不同基因突变类型的临床表型有较大差异:2例His348Gln及1例Arg304Trp纯合突变可表现为轻型和无症状,2例g.27_28delCT纯合、杂合突变分别表现为中型、无症状,4例双杂合突变分别表现为1例(Ser269Pro和Cys329Gly)无症状、2例(Arg304Trp和Cys329Gly与Arg277Cys和g.11520_11521insT)轻型、1例(Gln100Leu和His348Gln)中型.结论 中国汉族人群中存在导致F7基因缺陷的突变热点.遗传性凝血因子Ⅶ缺陷症患者的临床表型与FⅦ活性、F7基因突变类型无明显的相关性.     

Effect of Glycoprotein ⅢaT1565C Mutation on FAK Phosphorylation

Objective To study the effect of Glycoprotein Ⅲ aT1565C mutation on the phosphorylation of focal adhesion kinase (FAK). Methods The recombinants of pcDNA3.1( + ) Ⅱb and pcDNA3.1( + )Ⅲa or pcDNA3.1( + )ⅢaT1565C were transfected into CHO cells by Lipofectamine 2000. Cell lines expressing wild-type and mutational GP Ⅱ b/Ⅲ a were screened by G418. The constructed CHO cell lines were examed through flow cytometry (FCM) to detect the expression of CD41 and CD61. lmmunoprecipitation, Western blot and FCM were employed to detect the tyrosine and serine 910 phosphorylation of FAK in CHO cells stimulated by fibrinogen (Fbg). Results CD41 and CD61 were highly expressed in both CHO cell lines detected by FCM, which was 97.19% and 99.71%,respectively. The tyrosine phosphorylation of FAK was detected in CHO cells expressing wild-type and mutational GP Ⅱb/Ⅲa stimulated by Fbg for 90min, which was 16.24% and 20.44%, respectively. There was no serine 910 phosphorylation of FAK observed in both CHO cell lines stimulated by Fbg for 90min. However, serine 910 phosphorylation of FAK in the two cell lines was increased after 48h of stimulation to 34.89% and 73.84%, respectively.Conclusions CHO cell lines stably expressing wild-type GP Ⅱ b/Ⅲ a and mutational GPⅡb/ⅢaT1565C were constructed successfully. GPⅢaT1565C mutation could enhance the serine 910 and tyrosine phosphorylation of FAK. Increased phosphorylation of FAK may enhance GPⅡb/Ⅲ a-mediated outside-in signal transduction.     

The complete genomic sequence analysis of genotype 4 human astrovirus HASTVgz01 strain in Guangzhou

To analyze the genomic molecular structure and genotype of human astrovirus isolated from infant in Guangzhou of China, the primers were designed based on the genomic sequence of astrovirus from the GenBank and the target sequence were amplified by RT-PCR. Then the PCR-products were cloned to T vector and sequenced. The genomic nucleotide sequences were analyzed by the programs CLUSTAL W and DNASTAR. It was found that the full genomic length of HASTVgz01 strain was 6721 bp and the ORFs were 6558 bp. The 5' and 3' UTR were 82 and 81 nucleotides. The genome included 3 open reading frames (ORFs): ORF1a, ORF1b and ORF2. The 5'-terminal ORF1a started at nucleotide 83 and extended to nucleotide 2845. ORFlb (nt 2785 to nt 4332) overlaped ORFla by 61 nucleotides. The 3'-terminal ORF2 began at nucleotide 4325 and terminated at nucleotide 6640. ORF2 had 2316 nucleotides. Compared with other astrovirus sequences in GenBank, the homology of the amino acid sequence of ORF2 of HASTVgz01 strain with that of serotype 4 was 93% . Homology with other serotypes ranged from 61% to 70% . The complete nucleotide sequence of astrovirus HASTVgz01 strain isolated from Guangzhou in China was 6721 bp in length, GenBank accession NO. DQ344027. Comparing the ORF2 of astrovirus HASTVgz01 with the known sequences of types 1-8 the highest homology was serotype 4 (93%). Comparative sequence analysis of the HASTVgz01 ORF2 with the reported human astrovirus sequences revealed that the isolated astrovirus belongs to genotype (serotype) 4.     

Clinical and molecular genetic analysis for 7 patients from 5 pedigrees with 17α-hydroxylase/17, 20 lyase deficiency

Objective To investigate the clinical and genetic characteristics of 7 patients from 5 families with 17α-hydroxylase/17, 20 lyase deficiency (17OHD) and the CYP17A1 mutation in Chinese. Methods Clinical features and laboratory data were collected from 5 families with 17OHD. PCR direct sequencing was performed to screen the mutation of CYP17A1 gene of the patients. Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) and sequencing were performed to screen the mutations of CYP17A1 gene in 288 healthy individuals from Shandong province. Results Seven patients (5 of them were 46, XX; 2 were 46, XY) had typical clinical presentation of sexual infantilism, hypertension and hypokalemia. Hormone profile indicated decreased plasma cotisol and sex hormones, and elevated blood adrenocorticotrophic hormone (ACTH). TAC329AA and H373L in exon 6 and D487_ F489del in exon 8 were identified from the patients. One heterozygote for D487_F489del was identified in 288 healthy controls. Conclusion The TAC329AA and D487_F489del of the CYP17A1 gene were the most frequent mutations in Chinese with 17OHD. There might be certain frequency of heterozygotes for D487_ F489del in Chinese population.