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Development of Bead-Based Suspension Array Technology for the Diagnosis of Hereditary Hearing Loss

Development of Bead-Based Suspension Array Technology for th...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Liu Chang Zhang Liang Zhang Yan Ding Hongke Zhou Weiping Feng Defeng Zeng Yukun Liu Ling Yin Aihua Medical Genetic Centre Guangdong Women and Children Hospital Maternal and Children Metabolic-Genetic Key Laboratory Guangdong Women and Children Hospital

Objective Hearing loss is one of the most common birth defects and one of the most prevalent se nsorineural disorders. Given that hereditary hearing impairment exhibits incredible genetic hetero geneity, effective diagnostic methods are highly demanded by the clinical management. Method We developed a new diagnostic assay using the bead-based suspension array technolo gy(BBSAT), which can detect 20 point mutations in four genes of Asian prevalent hearing impair ment in parallel within 5 hours. More than 200 previously genotyped clinical samples were used t o determine the cut-off values and to validate the accuracy of new assay. Result In the BBSAT assay, suspended beads increase surface area to 3-D and more probes co ncentrated on beads. Besides, probes and targets mixed in the same mobile phase also generate better hybridization dynamics. With the multiplex PCR primer and probe design strategy, 20 point mutations in four genes of Asian prevalent hearing impairment can be simultaneously detected in a single assay. The results of BBSAT were completely in line with known genotypes, indicating th at the new diagnostic assay possesses excellent clinical sensitivity(100%) and specificity(100%) to identify genetic mutations of hearing loss. Conclusion The BBSAT is a rapid, accurate and cost-effective technology, suitable for molecular diagnosis of the hereditary hearing loss in large populations.

关键词： Development of Bead-Based Suspension Array Technology for the Diagnosis of Hereditary Hearing Loss

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Non-Invasive Pre-Implantation Aneuploidy Screening and Diagnosis of Beta Thalassemia IVSⅡ654 Mutation using Spent Embryo Culture Medium

Non-Invasive Pre-Implantation Aneuploidy Screening and Diagn...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Liu Weiqiang Liu Jian Qiao Du Hong Zi Ling Jia Wei Sun Xiao Fang Chen Dun Jin Third affiliated hospital of guangzhou medical university

Objective Cell-free nuclear DNA has been isolated from spent embryo culture medium. Whether these small amount of DNA can be amplified at the whole genome level and the concordance rate of karyotypes and specific alleles between biopsied cells and media has not been evaluated. The purpose of this study is to investigate whether cell-free nuclear DNA isolated from spent culture medium can be amplified at the whole genome level and to compare the concordance rate of karyotypes and specific alleles between biopsied cells and media. Method Seven couples were recruited and 88 donated embryos and their corresponding media were collected for whole genome amplification(WGA). The efficiency of WGA, and the concordance of chromosome status and the HBB gene IVSⅡ654 allele between biopsied cells and media were investigated using Next Generation Sequencing. Result After WGA, the DNA detection rate was 90.90% with a mean concentration of 26.15 ng/μl. The full chromosome concordance rate between biopsied cells and medium was 64.52%, and it increased to 90.00% for diploid blastocyst samples. Analysis of the mutated IVSⅡ654 locus and SNP linkage verified that the DNA present in the medium originated from embryonic cells. Conclusion We confirmed that nuclear DNA is present in spent culture medium, and that the majority of this DNA can be amplified for subsequent analysis. Our results showed that noninvasive embryo genetic testing at the chromosomal-level using medium can concordant to the biopsied cells, but it needs further optimized before use in clinical applications.

关键词： Non-Invasive Pre-Implantation Aneuploidy Screening and Diagnosis of Beta Thalassemia IVS Mutation using Spent Embryo Culture Medium

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DNA methylation mediates genotype and smoking interaction in the development of anti-citrullinated peptide antibody-positive rheumatoid arthritis

DNA methylation mediates genotype and smoking interaction in...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Liu Yun 复旦大学基础医学院

Objective Rheumatoid arthritis(RA) is a chronic autoimmune disease that leads to inflammation of the joints and surrounding tissues. Multiple factors, including interactions between genetic andenvironmental risks, are important in RA susceptibility. However, the underlying functional background is not fully understood. Epigenetic modifications, such as DNA methylation, can be influenced by both internal genetic and external environmental risk factors and are critical for phenotypic plasticity. Here, we tested the hypothesis that DNA methylation can integrate both genetic and environmental cues and mediate the interaction between genotype and smoking in the development of anti-citrullinated peptide antibody(ACPA)-positive RA. Method Using data from multiple cohorts, one significant interaction between genotype and smoking on DNA methylation was identified and replicated using a linear regression model. This interaction between genotype and smoking is a novel interaction impacting the risk of developing ACPA-positive in both a cohort composed of Caucasians and a cohort composed of Asian descent, evaluated by a linear regression model as well as by departure from the additivity of effects. Finally, we showed that DNA methylation mediates this gene-environment interaction on the risk of developing of ACPA-positive RA by a mediation analysis. Result We identified one significant gene-environment interaction between a SNP and smoking on DNA methylation, which has been shown previously to be associated with the risk of developing ACPA-positive RA. Additionally, we show that the gene-environment interaction represents a novel interaction that confers the risk of developing ACPA-positive RA in different ethnic groups and this gene-environment interaction is mediated through DNA methylation. Conclusion To the best of our knowledge, this is the first report showing that DNA methylation can mediate gene-environment interaction regarding the development of a common disease, thus

关键词： DNA methylation mediates genotype and smoking interaction in the development of anti-citrullinated peptide antibody-positive rhe

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PKLR polymorphism and gastric cancer risk

PKLR polymorphism and gastric cancer risk

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Bi Yan Zhang Sheng-jian Zhao Wan Yu Tao Li Xingwang He Lin Qiu Li-xin He Guang Bio-X Institutes Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education) Shanghai Jiao Tong University Shanghai Key Laboratory of Psychotic Disorders Shanghai Institute of Mental Health Shanghai Jiao Tong University Fudan University Shanghai Cancer Center Department of Oncology Shanghai Medical College Fudan University Department of Oncology Suzhou Science & Technology Town Hospital Institutes of Biomedical Sciences Fudan University Institute for Nutritional Sciences Shanghai Institutes of Biological Sciences Chinese Academy of Sciences Cancer Institute Collaborative Innovation Center for Cancer Medicine Fudan University Shanghai Cancer Center

Objective Gastric cancer(GCa) is a leading cause of cancer-related death with high incidence worldwide, particularly in China. There have been some genes found to be involved in the initiation and progression of GCa and PKLR is an emerging gene in malignant tumors. The PKLR rs3762272 A>G polymorphism showed association with GCa in seldom literature. We intend to further co nfirm the role of rs3762272 in GCa pathology. Method we performed a hospital-based case-control study recruiting 1115 cases and 1172 controls from a Chinese Han population. Result We found that the Gallele of rs3762272 was associated with areduced risk of GCa [GG vs. AA, OR = 0.62, 95% CI = 0.44-0.88;AG/GG vs. AA, OR = 0.83, 95% CI = 0.70-0.98;GG vs. A A/AG, OR = 0.66, 95% CI = 0.47-0.92]. And the statistical power still significant in subgroups of age, sex, drinking status, smoking status and tumor site respectively. Conclusion Our result in the present study suggest the mild protection role of PKLR rs3762272 in GCa in Chinese Han people.

关键词： PKLR polymorphism and gastric cancer risk

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Association study of GRM7 polymorphisms with major depressive disorder in the Chinese Han population

Association study of GRM7 polymorphisms with major depressiv...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Niu Weibo Huang Xiaoye Chen Shiqing Li Xingwang Wu Xi Cao Yanfei Zhang Rui Bi Yan Yang Fengping Wang Lu Li Weidong Xu Yifeng He Lin Yu Tao He Guang Bio-X Institutes Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education) Shanghai Jiao Tong University Institute for Nutritional Sciences Shanghai Institutes of Biological Sciences Chinese Academy of Sciences Shanghai Key Laboratory of Psychotic Disorders Shanghai Institute of Mental Health Shanghai Jiao Tong University Institutes of Biomedical Sciences Fudan University

Objective Major depressive disorder(MDD) is a severe and polygenic psychiatric disorder with hi gh heritability. There is an evidence that metabotropic glutamate receptor 7(GRM7) is associate d with schizophrenia, bipolar disorder(BP), attention deficit hyperactivity disorder(ADHD) and M DD. In this study, we investigated whether single nucleotide polymorphisms(SNPs) in GRM7 wer e associated with MDD. Method We carried out a case-control study that was to genotype four single nucleotide polymorp hisms(SNPs)(rs9814881, rs13353402, rs9870680 and rs1531939) in 568 major depressive disor der patients and 846 healthy controls of Chinese Han origin. Result The results showed that SNP rs9814881 in GRM7 displayed the significant genotype distri bution with MDD(P = 0.0287). Nevertheless, there was no association in allele frequencies. Conclusion In summary, the results of this study indicate that the GRM7 might be associated wit h major depressive disorder in Chinese Han population.

关键词： MDD Association study of GRM7 polymorphisms with major depressive disorder in the Chinese Han population

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A Novel Homozygous Missense Mutation and A Compound Heterozygous Frameshift Mutations of PDZD7 Gene Identified in Two Chinese Families with Nonsyndromic Hearing Loss.

A Novel Homozygous Missense Mutation and A Compound Heterozy...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Guan Jing Wang Dayong Wang Hongyang Xie Linyi Yang Ju Zhao Cui Lan Lan Yin Zifang Wang Qiuju Department of Otolaryngology-Head and Neck Surgery Chinese PLA Institute of Otolaryngology Chinese PL A General Hospital

Objective To investigate the genetic etiology of two non-consanguineous Chinese families with a utosomal recessive non-syndromic hearing loss(ARNSHL). Method Two Chinese ARNSHL families with two affected siblings were recruited in this study. Th e families were analyzed for exons, splicing sites and immediate flanking intron sequences by tar geted region capture and high throughput sequencing. Furthermore, 1751 normal Chinese individ uals served as controls and 122 Chinese families segregating apparent ARNSHL, who have been previously excluded common deafness genes, were ascertained for screening candidate mutation s. Result We identified a novel homozygous missense mutation(***66 Leu) and novel compound heterozygous frameshift mutations(***56 fs Ter24 and ***403 fs Ter36) in the both families. W e screened 1751 normal Chinese individuals and 122 unrelated Chinese families segregating app arent ARNSHL failed to detect the three confirmatory mutations. Conclusion We report novel biallelic mutations in PDZD, which is the first study to identify PDZD7 as the ARNSHL-associated gene in Chinese population. Our finding could expand the pathoge nic spectrum and strengthens the role of PDZD7 biallelic mutations cause ARNSHL.

关键词： A Novel Homozygous Missense Mutation and A Compound Heterozygous Frameshift Mutations of PDZD7 Gene Identified in Two Chinese Families with Nonsyndromic Hearing Loss

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Molecular analysis of 12 Pakistani families with non-syndromic or syndromic hearing loss

Molecular analysis of 12 Pakistani families with non-syndrom...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Wang Rongrong Han Shirui Khan Amjad Zhang Xue Institute of Basic Medical Sciences Chinese Academy of Medical Sciences School of Basic Medicine Peking Union Medical College China Medical University

Objective To investigate the causative genetic mutations in 12 Pakistani families with non-syndro mic or syndromic hearing loss Method Mutations in the commonest causative gene GJB2 were pre-excluded by Sanger sequen cing. Then, targeted next-generation sequencing or whole-exome sequencing was carried out to analyze the genomic DNA samples from the 11 prob ands with hearing loss. Sanger sequencing was performed to verify all identi?ed variants. Result We found pathogenic or likely to be pathogenic mutations in all 12 families, including six k nown mutations in GJB2, SLC26 A4, LHFPL5, or USH2 A and eight novel mutations in ESPN, MY O7 A, LRTOMT, PCDH15, USH2 A or EPS8 L2. Notably, four compound heterozygous mutations i n the MYO7 A and USH2 A genes were detected in two consanguineous families, respectively. In addition, the novel frameshift mutation in EPS8 L2 was first documented in Pakistan. Conclusion Our study has increased the spectrum of mutations associated with hearing loss in t he Pakistani population. In addition, our study highlights the possibility of rare occurrences of com pound heterozygous mutations in consanguineous families.

关键词： GJB Molecular analysis of 12 Pakistani families with non-syndromic or syndromic hearing loss

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Alternatively spliced products lacking exon 12 dominate the expression of fragile X mental retardation 1 gene in human tissues

Alternatively spliced products lacking exon 12 dominate the ...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Fu Xianguo Department of Medical Genetics Fuzhou General Hospital

Objective Fragile X mental retardation 1 gene(FMR1) expression is associated with fragile X syn drome(FXS) and exhibits several splicing products. However, the proportion of spliced isoforms t hat are expressed in different tissues remains *** In the present study, long-chain reverse transcription-polymerase chain reaction with a T cloning-sequencing method was conducted in order to analyze the entire coding region of the FM R1 gene in human tissues. In particular, FXS-associated tissues were analyzed, including the brai n and testis. Result Twenty alternatively spliced isoforms were observed among 271 recombinants, including six novel ones. The isoform that consisted of the entire FMR1 coding region(ISO1) accounted for a small proportion of all isoforms. Isoforms lacking exon 12 were the most abundant. In particular, spliced isoforms ISO7 and ISO17 were the most abundant. However, their relative abundance va ried between the peripheral blood cells, and the testis and brain tissues. Bioinformatic analyses s uggested that exon 12 may be the sole exon undergoing positive selection. The results of the pre sent study suggested that the mechanisms underlying alternative splicing(AS) of the FMR1 gene may be more complex. Furthermore, the functions of alternatively spliced products lacking exon 12 require further investigation. Conclusion The results of the present study provide novel insights into the association between AS and the structure and function of the FMR1 gene.

关键词： Alternatively spliced products lacking exon 12 dominate the expression of fragile X mental retardation 1 gene in human tissues

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Effects of G6PD activity inhibition on the mechanical properties and cytoskeletons of cervical cancer cells

Effects of G6PD activity inhibition on the mechanical proper...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Fang ZiShui Jiang ChengRui Feng Yi Chen RiXin Lin XiaoYing Zhang ZhiQiang Han LuHao Chen XiaoDan Li HongYi Guo YiBin Jiang WeiYing Sun Yat-sen University

Objective Glucose-6-phosphate dehydrogenase(G6 PD) deficiency has been revealed to be involved in the efficacy to anti-cancer therapy but the mechanism remains unclear. In our previous study, we haveobserved increased ROS generation caused by the inhibition of G6 PD activity using Dehydroepiandrosterone(DHEA) and sh RNA technology. We aimed to investigate the further anti-cancer mechanism of G6 PD deficiency. Method In the present study, Peak Force QNM Atomic Force Microscopy was used to assess the changes of topography and biomechanical properties of cells and detect the effects on living cells in a natural aqueous environment. Moreover, Laser Scanning Confocal Microscope was used to observe the alterations in cytoskeleton to explore the involved mechanism. Result When G6 PD was inhibited by DHEA or sh RNA interference, the abnormal Young‘s modulus and increased roughness of cell membrane were observed in He La cells. Simultaneously, the inhibition of G6 PD activity caused the disorganization of microfilaments and microtubules of cytoskeletons. Conclusion Our results indicated the anti-cervix cancer mechanism of G6 PD deficiency may be involved with the abnormal reorganization of cell cytoskeleton and abnormal biomechanical properties caused by the increased ROS due to the inhibition of G6 PD activity. The results suggested suppression of G6 PD may be a promising strategy in developing novel therapeutic methods for cervical cancer.

关键词： Effects of G6PD activity inhibition on the mechanical properties and cytoskeletons of cervical cancer cells

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A novel insertion mutation in the ATP7A gene associated with delayed infantile onset of Menkes disease

A novel insertion mutation in the ATP7A gene associated with...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Li Wugao Yan Tizhen Tang Ning Li Zhetao Huang Jiwei Tan Jianqiang Pan Lizhen Cai Ren Key Laboratory of birth defects prevention and control Department of Medical Genetics Liuzhou Municipal Maternity and Child Healthcare Hospital

Objective Determining the relationship between clinical phenotype and genotype in genetic disea ses is important in clinical practice. In general, frameshift mutations are expected to produce pre mature termination codons, leading to production of mutant transcripts destined for degradation b y nonsense-mediated decay. In X-linked recessive diseases, male patients with frameshift mutati ons typically have a severe or even lethal phenotype. Method We report a case of a 22-month-old boy patient with Menkes disease, an X-linked recess ive copper metabolism disorder, caused by mutations in the ATP7 A copper transporter gene. His clinical presentation involved unexpecte dly late onset and milder symptoms. Result His genomic DNA showed a de novo insertion mutation(c.1036 ins T) in ATP7 A gene, pred icting a translational frameshift and premature stop codon, and a classic severe phenotype. Char acterization of his ATP7 A m RNA showed no abnormal splicing. Conclusion We speculated that translation re-initiation could occur downstream of the premature termination codon, and produce a partially functional ATP7 A protein. Study of the patient‘s fibrobl asts found no evidence of translation reinitiation, however the possibility remains that this pheno menon occurred in neural tissues and influenced the clinical phenotype.

关键词： ATP A novel insertion mutation in the ATP7A gene associated with delayed infantile onset of Menkes disease

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