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Using an exon microarray to identify a global profile of gene expression and alternative splicing in K562 cellsexposed to sodium valproate

Using an exon microarray to identify a global profile of gen...

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广东省遗传学会第九届代表大会暨学术研讨会

作者： Zhang XZ Yin AH Zhu XY Ding Q Wang CH Chen YX Prenatal Diagnosis Centre Guangdong Women and Children Hospital

To investigate the effect of valproate treatment on the K562 cell line,a model for chronic myelogenous leukaemia,the growth and survival of the K562 cell line were investigated using the Annexin-V/PI dual staining method,and global profiles of gene expression and alternative splicing in K562 cellswere assessed using exon microarrays.A significant increase in cell apoptosis was observed in valproate-exposed K562 cells using flow cytometry.A total of 628 transcripts were identified as being significantly differentially *** number of genes demonstrating increased expression levels was greater than the number of genes demonstrating decreased expression levels(445genes vs.183 genes,respectively).The significant enrichment analysis of GO terms for the differentially expressed genes revealed that these genes are involved in many important biological processes such as *** of the genes observed to be differentially expressed that might be involved in apoptosis were selected to undergo qRT-PCR *** total,198 candidates of alternative splicing variants were *** them,three alternative splicing events were selected for validation,and CBLC and TBX1 were confirmed to be alternatively spliced by semi-nested *** conclusion,valproate exposure facilitated cell apoptosis,altered mRNA expression and alternative splicing events in the K562 cell line.

关键词： profile microarray Using an exon microarray to identify a global profile of gene expression and alternative splicing in K562 cellsexposed to sodium valproate

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Non-invasive prenatal diagnosis of trisomy 21 by reverse transcriptase multiplex ligation-dependent probe amplification

Non-invasive prenatal diagnosis of trisomy 21 by reverse tra...

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广东省遗传学会第九届代表大会暨学术研讨会

作者： Deng YH Yin AH He Q Chen JC He YS Wang HQ Li M Chen HY Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Background:Obtaining fetal DNA or RNA by either chorionic villus sampling(CVS) or amniocentesis is currently,the gold standard ***,these invasive procedures carry risk of miscarriage.A reliable method for non-invasive prenatal diagnosis(NIPD) has long been sought to reduce the risk of ***:Cell-free fetal RNA was extracted from the plasma of peripheral blood from 121 women9-20 weeks of *** single nucleotide polymorphism(SNP) loci in PLAC4 gene were analyzed by reverse transcriptase multiplex ligation-dependent probe amplification(RT-MLPA),followed by capillary *** analysis was used for confirmation of prenatal diagnosis of trisomy ***:Of 121 samples,23 were diagnosed with trisomy 21,87 with normal ploidy,nine had all five SNP loci homozygous and two had one heterozygous SNP *** with karyotype analysis,the diagnostic sensitivity and specificity of RT-MLPA were 92%and 100%,***:RT-MLPA is a convenient and reliable method for the diagnosis of trisomy *** have shown that this method has good specificity,high sensitivity,and high throughput,making this technique applicable for NIPD in clinical practice.

关键词： Non-invasive prenatal diagnosis of trisomy 21 by reverse transcriptase multiplex ligation-dependent probe amplification

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Genetic diagnosis of Chinese Han patients with Polycystic Kidney Disease

Genetic diagnosis of Chinese Han patients with Polycystic Ki...

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广东省遗传学会第九届代表大会暨学术研讨会

作者： Zishui Fang Shiyan Xu Weiying Jiang Department of Medical Genetics Zhongshan School of MedicineSun Yat-sen University Shen Zhen People's Hospital

Polycystic kidney disease(PKD) includes autosomal dominant polycystic kidney disease(ADPKD) and autosomal recessive polycystic kidney disease(ARPKD),which has always been associated with a high prevalence *** clinical diagnosis of the disease has numerous limitations,genetic testing has proved to be useful in the early diagnosis of asymptomatic patients,with or without family *** molecular methods involved prenatal as well as pre-symptomatic ***,the clinical is difficult to distinguish between fetal ADPKD and ARPKD,particularly few reports of the relevant PKD1,PKD2,and PKHD1 genes of diagnosis in the Chinese *** diagnosis of the disease in at-risk individuals is complicated due to the structural complexity of PKD1 gene and the high diversity of the *** aimed to revolutionize the diagnosis protocol of PKD to provide earlier diagnosis and an timely *** the meantime,we explored preimplantation genetic diagnosis of PKD *** results indicated that devising and running a systematic operating procedure in identifying the pathogenic mutations in PKD1,PKD2 and PKHD1 genes could offer great promise in the diagnosis and treatment of ADPKD and ARPKD among *** direct sequencing and DHPLC combining prediction of the pathogenicity of missense variants,we screened these genes for mutations in 13 families with PKD,and found 7novel mutations including 4 mutations in PKD1 gene and 3 mutations in PKHD1 gene,which enrich spectrum of mutations Chinese Han population.

关键词： Polycystic kidney disease(PKD) PKD1 PKD2 PKHD1 DNA sequencing DHPLC Novel mutations Pathogenicity prediction PGD

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A sensitive time-resolved fluoroimmunoassay for determination of median levels of pregnancy-associatedplasma protein a in pregnant women in China

A sensitive time-resolved fluoroimmunoassay for determinatio...

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广东省遗传学会第九届代表大会暨学术研讨会

作者： Yu T Gao S Yin A Tang Y Wu Y Li L Li M Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Pregnancy-associated plasma protein A(PAPP-A) is an important serum marker for first trimester *** weekly median value varies with ethnicity.A one-step time-resolved fluoroimmuno assay(TRFIA) using two monoclonal antibodies against PAPP-A with Eu(3+) chelates as labels has been *** the assay described here,we evaluated 5,301 normal serum samples from Chinese women at 7-13 weeks of *** detection limit using this assay was 1.2 mlU/L,and the maximum detection range was up to 10,000 mlU/*** intra-assay and inter-assay coefficients of variation were <3.0%and <5.0%,respectively,and the mean recovery rate was 98.0%.PAPP-A concentrations measured in 516 maternal serum samples correlated well with those obtained by Dissociation-Enhanced Lanthanide Fluorescent Immunoassay(DELFIA) PAPP-A assay(r=0.988,P<0.001).The medians for 7-13 weeks of maternal serum PAPP-A were higher in the women from China compared to reports from other *** present assay possesses accuracy and high sensitivity and exhibits great potential for the clinical analysis of *** investigation on themedian concentrations of PAPP-A will help establish reference values that are specific for China and study the importance of ethnic factors in biochemical screening.

关键词： A sensitive time-resolved fluoroimmunoassay for determination of median levels of pregnancy-associatedplasma protein a in pregnant women in China

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莱茵衣藻酰基辅酶A合成酶功能验证及其调控脂肪酸代谢的研究

莱茵衣藻酰基辅酶A合成酶功能验证及其调控脂肪酸代谢的研究

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广东省遗传学会第九届代表大会暨学术研讨会

作者：宋燕子贾彬林柏成胡章立黄瑛深圳市海洋生物资源与生态环境重点实验室深圳市海洋藻类开发与应用工程实验室深圳大学生命科学学院

能源微藻胞内油脂提取工艺复杂,过程能耗极高,若能实现微藻油脂的胞外分泌,必将促进能源微藻的产业化进程。Yasunari M和Michael S等人分别通过敲除脂肪酸酰基辅酶A合成酶基因的方法先后在酿酒酵母Saccharomyces cerevisiae中实现了油... 详细信息

能源微藻胞内油脂提取工艺复杂,过程能耗极高,若能实现微藻油脂的胞外分泌,必将促进能源微藻的产业化进程。Yasunari M和Michael S等人分别通过敲除脂肪酸酰基辅酶A合成酶基因的方法先后在酿酒酵母Saccharomyces cerevisiae中实现了油脂的胞外分泌,使微藻油脂的胞外分泌研究成为可能。基于上述背景,本研究以莱茵衣藻(chlamydomonas reinhardtii)为对象旨从酰基辅酶A合成酶的角度对微藻油脂的代谢进行研究,为今后进一步研究微藻油脂的胞外分泌奠定基础。本研究利用生物信息学的方法从莱茵衣藻基因组数据库中预测出两个莱茵衣藻ACS基因,再通过RT-PCR从莱茵衣藻CC-849中克隆它们并分别命名为cracsl,cracs2,然后用多重比对和分子进化树对其编码蛋白进行功能预测。为了进一步探索cracs1,cracs2的功能,本实验分别在酿酒酵母YB525和莱茵衣藻CC-849中对其进行体外和体内功能研究。在体外功能研究中,本实验将cracs1,cracs2连接到酵母表达载体pYES2上,通过电转化分别将它们转入ACS缺陷型酵母,再以不同碳链长度脂肪酸作为唯一碳源分别培养阳性酵母转化子,探究CrACS1和CrACS2的功能;在体内功能研究中,本实验采用荧光定量PCR的方法检测缺氮处理CC-849中cracs1,cracs2基因表达情况,进一步分析CrACS1和CrACS2与油脂代谢的关系。本实验首次从莱茵衣藻中克隆了两个cracs基因,其中cracs1长2004bp,基因序列与预测序列完全一致,cracs2长2016bp,基因序列比预测序列多出399bp,分析发现基因中多出的序列位于莱茵衣藻基因组的gap区域。多重比对结果表明CrACS1和CrACS2均含有中长链ACS的两个保守区:AMP-binding区和FACS区,分子进化树预测CrACS1和CrACS2属于长链ACS,CrACS1参与脂肪酸的合成,CrACS2参与脂肪酸的β氧化和脂肪酸的转运。后续莱茵衣藻ACS的体外功能研究证实CrACS1和CrACS2均属于酰基辅酶A合成酶家族,能恢复缺陷型酵母YB525的酰基辅酶A合成酶活性,活化外源脂肪酸,参与油脂的合成与代谢;体内功能实验则发现随着缺氮时间的延长cracs1的表达量总体呈上升趋势,而cracs2的表达量却逐渐下降,缺氮72h时,cracs2的相对表达量下降6.7倍,此结果进一步表明CrACS1可能参与脂肪酸的合成,CrACS2可能参与脂肪酸的β氧化。CrACS1和CrACS2与莱茵衣藻油脂分泌之间的关系也在进一步研究中。

关键词：莱茵衣藻酰基辅酶A合成酶脂肪酸代谢油脂分泌

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The carrier rate and mutation spectrum of genes associated with hearing loss in South China hearing femalepopulation of childbearing age

The carrier rate and mutation spectrum of genes associated w...

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广东省遗传学会第九届代表大会暨学术研讨会

作者： Yin A Liu C Zhang Y Wu J Mai M Ding H Yang J Zhang X Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Background:Given that hearing loss occurs in 1 to 3 of 1,000 live births and approximately 90 to 95 percent of them are born into hearing families,it is of importance and necessity to get better understanding about the carrier rate and mutation spectrum of genes associated with hearing impairment in the general ***:7,263 unrelated women of childbearing age with normal hearing and without family history of hearing loss were tested with allele-specific PCR-based universal *** genetic testing were provided to the spouses of the screened *** those couples at risk,multiple choices were provided,including prenatal ***:Among the 7,263 normal hearing participants,303 subjects carried pathogenic mutations included in the screening chip,which made thecarrier rate 4.17%.Of the 303 screened carriers,282 harbored heterozygous mutated genes associated with autosomal recessive hearing loss,and 95 spouses took further genetic tests.8 out of the 9 couples harbored deafness-causing mutations in the same gene received prenatal ***:Given that nearly 90 to 95 percent of deaf and hard-of-hearing babies are born into hearing families,better understanding about thecarrier rate and mutation spectrum of genes associated with hearing impairment in the female population of childbearing age may be of importance incarrier screening and genetic counseling.

关键词： The carrier rate and mutation spectrum of genes associated with hearing loss in South China hearing femalepopulation of childbearing age

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Genetic counseling and prenatal diagnosis for hereditary hearing loss in high-risk families

Genetic counseling and prenatal diagnosis for hereditary hea...

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广东省遗传学会第九届代表大会暨学术研讨会

作者： Yin A Liu C Zhang Y Wu J Mai M Ding H Yang J Zhang X Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Background:Genetic counseling and prenatal diagnosis are very necessary and accurate to detect hereditary hearing loss,especially in *** diagnosis is testing for diseases or conditions in fetuses before born,which gives parents the chance to prepare psychologically,financially and medically for the probable health and educational needs of the affected ***:54 unrelated families with children affected with non-syndromic sensorineural hearing loss were enrolled in the study and received geneticanalysis with microarray and DNA sequencing *** counseling was provided to each participating families,and prenatal diagnosiswas given to those at risk and would like to know their fetuses' genotypes and probable hearing ***:Half the cases in the present study were diagnosed with confirmed pathogenic mutations and clear inheritance *** receivinggenetic counseling,24 carrier couples with pathogenic mutations chose to proceed prenatal diagnosis,the results of which were in accordance with the pregnancy *** prenatally detected to be monoallelic mutation carriers and those harbored neither deafness-causing mutations form their parents passed newborn hearing screening and six-month follow-ups,while neonates prenatally detected to be carriers of diallelic or compound heterozygous mutations developed hearing loss after ***:With appropriate genetic counseling and support services provided,the genetic testing and the prenatal diagnosis of hearing losswere valued by carrier couples for the information provided for future family planning and probably the preparation for the health and educational needs of the affected neonates.

关键词： Genetic counseling and prenatal diagnosis for hereditary hearing loss in high-risk families

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A prenatal missed diagnosed case of submicroscopic chromosomal abnormalities by karyotyping:the clinical utility of array-based CGH in prenatal diagnostics

A prenatal missed diagnosed case of submicroscopic chromosom...

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广东省遗传学会第九届代表大会暨学术研讨会

作者： Yin A Lu J Liu C Guo L Wu J Mai M Zhong Y Zhang X Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Background:Array-based comparative genomic hybridization possesses a number of significant advantages over conventional cytogenetic and other molecular cytogenetic techniques,providing a sensitive and comprehensive detection platform for unexpected imbalances in the genome *** Presentation:The newborn proband,demonstrated with craniofacial dysmorphism and multiple malformations,was born to a family with spontaneous *** pregnancy was uneventful,except the prenatal ultrasound examination showed an increased nuchal translucency at 12(+) weeks of *** revealed an apparently normal karyotype,and the couple decided to continue the ***-basedCGH analysis was applied to the affected infant,identified a combination of 18 p deletion and 7q *** study indicates that the unbalanced translocation was inherited from a balanced translocation carrier ***:In review of the case,several overlooked points leading to the missed diagnosis should be discussed and certain quality control strategies should be adopted to avoid similar problems in the ***-based CGH and karyotyping techniques are complemented by diverse detection spectrum and resolutions,and a combination of these methods could help providing optimal genetic *** that the array-CGHanalysis will not introduce additional risk to patients,it is reasonable to recommend those already undergoing invasive testing should take array-basedCGH as an adjunct to conventional cytogenetic tests and other molecular cytogenetic analysis.

关键词： Array-based CGH Prenatal diagnostics Submicroscopic chromosomal abnormalities

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应用EGS引导核酶P高效抑制HCMV临床病毒株UL148基因表达

应用EGS引导核酶P高效抑制HCMV临床病毒株UL148基因表达

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广东省遗传学会第九届代表大会暨学术研讨会

作者：胡兢晶苏海浩丁俊彩郭媛媛谢斌华伍苑宾密英鸽王波广东省儿童医院·广东省妇幼保健院儿科

背景目前,婴幼儿HCMV感染的发病率逐年升高,婴儿先天性感染往往导致先天畸形、耳聋、智力障碍等严重后遗症,甚至死亡。有效防治或阻断HCMV先天感染对提高儿童健康水平、生活质量方面具有重大意义。随着病毒部分分子机制的明确,目前治疗H... 详细信息

背景目前,婴幼儿HCMV感染的发病率逐年升高,婴儿先天性感染往往导致先天畸形、耳聋、智力障碍等严重后遗症,甚至死亡。有效防治或阻断HCMV先天感染对提高儿童健康水平、生活质量方面具有重大意义。随着病毒部分分子机制的明确,目前治疗HCMV感染已取得了重大成效,但HCMV耐药株的出现及抗HCMV药物的副作用,使其在儿童和孕妇感染者中的使用受到一定的限制。防治婴儿HCMV感染尚无有效策略,阻断母婴传播,降低婴儿先天性HCMV感染仍是世界性难题。目的探讨外部引导序列(externa1 guide sequencees,EGSs)基因沉默技术对人巨细胞病毒临床病毒株特有基因UL148表达的抑制作用,以探索抗HCMV治疗的新途径,为开发新型反义核酸药物奠定基础。方法细胞外研究:应用RNA structure技术模拟UL148 RNA二级结构,选择二级结构相对简单的区域,以第362位为切割位点设计并合成DNA性质EGS362。体外扩增UL148基因及M1RNA基因,克隆至T7启动子下游,体外转录UL148 RNA及M1RNA,其中UL148 RNA以P标记。将P标记的UL148RNA及M1RNA混合至体外反应液中,进行EGS的引导下进行细胞外切割反应。细胞内研究:扩增UL148基因克隆至pEGFP-N1载体,构建UL148真核表达载体。将UL148真核表达载体转染Hela细胞,构建稳定表达UL148基因的细胞株,经荧光显微镜观察及RT-PCR的方法鉴定UL148 RNA在Hela细胞内的表达。将EGS362转染稳定表达UL148基因的Hela细胞株,分别以荧光显微镜及荧光定量PCR分析EGS362在真核细胞内对UL148表达的沉默效果。结果细胞外研究:经RNA structure技术体外模拟EGS362可于UL148 RNA形成类似M1RNA天然作用底物的径环结构,易于M1RNA的识别及切割。在细胞外获得UL148 RNA及M1RNA,经体外切割反应,可获得与预期大小相符的切割条带,证实DNA性质EGS362可特异性地切割UL148 RNA,其切割位点准确,与预期相符。细胞内研究:经UL148真核表达载体转染Hela细胞,经荧光显微镜观察可观察到绿色荧光,应用RT-PCR鉴定UL148基因在真核细胞中稳定表达,证实稳定表达UL148的Hela细胞系构建成功。分别以50nM、100 nM、200 nM、400 nM EGS362转染Hela细胞后,以荧光显微镜观察,绿色荧光蛋白较前明显下降,经荧光定量PCRUL148分析,其表达率下降分别为74.9%、78.6%、81.1%、87%。结论EGS362能准确的切割目标基因UL148 RNA,其准确性可精确到单个碱基的水平;经体外筛选出的EGS362能进一步在细胞内沉默UL148基因的表达;EGS362的沉默效率与其浓度成正相关;EGS362发展成为UL148的理想沉默工具;该研究也为明确该基因的功能奠定基础。

关键词：人巨细胞病毒外部引导序列 UL148 临床病毒株核酶P

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Continuous Glucose Monitoring Effects on Maternal Glycemic Control and Pregnancy Outcomes in Patients with Gestational Diabetes Mellitus:A Prospective Cohort Study

Continuous Glucose Monitoring Effects on Maternal Glycemic C...

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广东省遗传学会第九届代表大会暨学术研讨会

作者：余凡广东省妇产医院·广东省妇幼保健院

Clinical evidences on the consequential effects of continuous glucose monitoring(CGM) on pregnancy outcomes in women with GDM are scarcely *** To evaluate the effectiveness of CGM on maternal glycemic control and pregnancy outcomes in patients with ***:340 Chinese pregnant women with GDM were allocated to either the routine care group(n= 190) or the CGM group(n=150).Design and Setting:This was a prospective cohort study in Department of Obstetrics of GuangDong Women and Children Hospital in *** started in April 2011 and stopped in August ***:A 72 h continuous glucose monitoring system(CGMS) was used as a supplementary tool forglucose monitoring in the CGM *** Outcome Measurements The parameters of glycemic variability included:mean blood glucose(MBG);the standard deviation(SD) of blood glucose(SDBG);mean amplitude of glycemic excursions(MAGE) and the mean of daily differences(MODD).The maternal outcomes(preeclampsia and cesarean delivery) and composite neonatal outcomes were *** The SDBG,MAGE and MODD values were significantly lower in the CGM group compared to those of the routine care group(P <0.001).Subjects in the CGM group were at lower risk of preeclampsia and primary cesarean delivery compared with the routine care group(P < 0.05).The mean infant birth weight of women in the CGM group was lower than infants of women in the routine care group(P < 0.001).The MAGE was associated with birth weight(p= 0.196,P < 0.001),and it was an independent factor for preeclampsia(OR,3.66;95%CI 2.16- 6.20) and composite neonatal outcome(OR,1.34;95%CI 1.01-1.77).Conclusions The use of supplementary CGM combined with routine antenatal care can improve the glycemic control andpregnancy outcomes of patients with GDM.

关键词： GDM Continuous Glucose Monitoring Effects on Maternal Glycemic Control and Pregnancy Outcomes in Patients with Gestational Diabetes Mellitus:A Prospective Cohort Study

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