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MOLECULAR EPIDEMIOLOGY INVESTIGATION OF THALASSEMIA IN CHENGDU REGION,SICHUAN PROVINCE,SOUTHWEST CHINA

MOLECULAR EPIDEMIOLOGY INVESTIGATION OF THALASSEMIA IN CHENG...

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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： Xia Yu Li-ye Yang Hui-Tian Yang Cheng-gui Liu Deng-cheng Cao Wei Sheng Min Lin Department of Medical Laboratory Chengdu Women's & Children's Central Hospital Centeral Laboratory Chaozhou Central Hospital Affiliated to Southern Medical University

Thalassemia is the most common inherited diseases in southern ***,this disorder is usually ignored by health system in Sichuan Province due to lack of epidemiological *** provide basic epidemiology information for thalassemia screening,genetic counseling and prenatal diagnosis（PND） in Chengdu region,a total of3,262 healthy subjects were screened by completed blood cell（CBC） count,reverse dot blot（RDB） gene *** results presented that heterozygote frequency of thalassemia was 3.31%（108/3262）,of which 2.21%（72/3262） carried α-thalassemia and 1.16%（38/3262） were β-thalassemia *** types of a-thalassemia mutation,the most prevalent being the-（68.06%）,followed by-α（rightward）(25%),Hb Quong Sze（HBA2:c.377T>C）(4.17%) and-a37（leftward）(2.78%).The six point mutations identified in the β-thalassemia carriers were:HBB:c.316-197C>T（34.21%）,HBB:c.52A>T（28.95%）,HBB:c.l2629delCTTT（23.68%）,HBB:c.-78A>G（7.89%）,HBB:c.-79A>G（2.63%） and HBB:c.845insC（2.63%）.This was the first detail molecular epidemiological survey of thalassemia in Chengdu region of Sichuan province,People's Republic of China（PRC）.

关键词： Thalassemia Molecular epidemiology survey Chengdu

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The prevalence and molecular spectrum of α-and β-globin gene mutations in 14,332 families of Guangdong Province,China

The prevalence and molecular spectrum of α-and β-globin ge...

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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 Li B Luo M Xu L Wu L Zhang L Ma Y Chen T Gao S Liang J Guo H Qin D Wang J Yuan T Wang Y Huang WW He WF Zhang Y Liu C Xia S Chen Q Zhao Q Zhang X Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Objective:To reveal the familial prevalence and molecular variation of α- andβ-globin gene mutations in Guangdong ***:A total of 40,808 blood samples from 14,332 families were obtained and analyzed for both hematological and molecular ***:A high prevalence of a- and P-globin gene mutations ***,17.70%of pregnant women,15.94%of their husbands,16.03%of neonates,and 16.83%of couples(pregnant women and their husbands) were heterozygous carriers of α- or β-*** regions with the highest prevalence were the mountainous and western regions,followed by the Pearl River Delta;the region with the lowest prevalence was *** total familial carrier rate(both spouses were α- or β-thalassemia carriers) was 1.87%,and the individual carrier rates of α-andβ-thalassemia were 1.68%and 0.20%,*** total rate of moderate-to-severe fetal thalassemia was 12.78%among couples in which both parents were ***:There was a high prevalence of α-and β-thalassemia in Guangdong *** study will contribute to the development of thalassemia prevention and control strategies in Guangdong Province.

关键词： The prevalence and molecular spectrum of globin gene mutations in 14,332 families of Guangdong Province,China

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Screening significantly hypermethylated genes in fetal tissues compared with maternal blood using amethylated-CpG island recovery assay-based microarray

Screening significantly hypermethylated genes in fetal tissu...

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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 Zhang X Wu J Du L He T Zhang X Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Background:The noninvasive prenatal diagnosis procedures that are currently used to detect genetic diseases do not achieve desirable levels of sensitivity and ***,fetal methylated DNA biomarkers in maternal peripheral blood have been explored for the noninvasive prenatal detection of genetic ***,such efforts have covered only chromosomal aneuploidy,and fetal methylated DNA biomarkers in maternalwhole blood for detecting single-gene diseases remain to be ***:To address this issue,we systematically screened significantly hypermethylated genes in fetal tissues and compared them with maternalperipheral blood potential in an attempt to detect fetal genes in maternal peripheral ***,the methylated-CpG island recovery assay combined with a CpG island array was performed for four fetus-toward placental tissues and the corresponding maternal peripheral ***,direct bisulfite sequencing and combined bisulfite restriction analysis(COBRA) were carried out to validate the methylation status of the hypermethylated genes that were identified by the microarray ***:Three hundred and ten significantly hypermethylated genes in the placental tissues were detected by *** the top 15 hypermethylated genes detected by microarray,two were selected for sequencing validation in placental tissue and chorionic villus samples and four were selected for COBRA validation in four placental tissues,ten amniotic fluids and five chorionic villus *** six selected genes were confirmed to be hypermethylated in placental tissue and chorionic villus samples,but methylation of the genes could not be detected in the amniotic ***:Of the many hypermethylated genes and methylation sites that were found in the fetal tissues,some have great potential to be developed into molecular markers for noninvasive prenatal diagnosis of monogenic *** clinical studies are warranted to confirm these

关键词： microarray Screening significantly hypermethylated genes in fetal tissues compared with maternal blood using amethylated-CpG island recovery assay-based microarray CpG

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Determination of median levels of the free beta subunit of human chorionic gonadotropin in women from China's mainland using a new time-resolved fluoroimmunoassay

Determination of median levels of the free beta subunit of h...

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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： Tang YP Wu YS Yin AH Xu WW Yuan PH Li M Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Background:The free beta subunit of human chorionic gonadotropin(free beta-hCG) is an important serum marker for biochemical *** weekly median value varies with *** of the fluorometers for lanthanide chelates are designed for the detection of signals from europium(Eu(3+)) chelates ***:We developed a two-site,one-step assay using two monoclonal antibodies(MAbs) against free beta subunit and beta subunit with Eu(3+) chelates as *** the present assay,we evaluated 24,634 normal serum samples in Chinese pregnant women during 8-20 weeks of ***:The detection limit using this assay was <0.05 ng/*** within-run and between-run imprecision was <6.0%and 7.0%using control *** beta-hCG concentrations measured using the current assay in 999 maternal serum samples correlated well with those obtained by samarium(Sm(3+))-labeled DELFIA free hCGbeta assay(r=0.987).The medians for 8-20 weeks for maternal serum free beta-hCG were higher in the women from China's mainland compared to reports from other ***:The present assay is suitable for use in biochemical screening of women in China's *** study on the median concentrations of free beta-hCG will help establish reference values that are specific for ethnic populations from the Chinese *** will be useful for studying the importance of ethnic factors in biochemical screening.

关键词： hCG Determination of median levels of the free beta subunit of human chorionic gonadotropin in women from China's mainland using a new time-resolved fluoroimmunoassay

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Genotype of α and β-globin genes and the porpolymorphism of β-globin gene in Cantonese

Genotype of α and β-globin genes and the porpolymorphism o...

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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： Liu Ling

Objective Aim to provide an effective genetic testing for thalassemia(thal),we analyzed genotype a and P-globin,as well as the polymorphism of P-globin gene in Cantonese in recent *** Using single-tube complex PCR to detect deletional a-thal,reverse dot blotting(RDB)/PCR to detect 3 kinds undeletionalα-thal——αaCS,αQS,aWSand 18 kinds P-thal mutations which are common seen among Chinese population,454 cases from Guangdong were given thal genotype genetic *** the 454 cases,142 cases were selected to study the polymorphismsingle nucleotide polymorphisms(SNPs) of p-globin gene through denaturing high-performance liquid chromatography(DHPLC) combining the whole gene *** For 438 cases were diagnosed thalassemia,246 cases for a-thal,164 cases for P-thal,28 cases a-thal compounding β-*** 246 a-thal cases,deletions,the dominant mutations,257 cases were diagnosed αα/--SEA,20 cases αα/-α3.7,9cases αα/-α*** 164 β-thal cases,β-thal heterozygotes accounted for 92.7%(152/164),of which the main genotypes are CD41-42,IVS-Ⅱ-654,-28,CD17,dual heterozygotes and homozygotes accounted for 4.9%(8/164)and 2.4%(4/164) *** result of P-globin gene screening via DHPLC combining with sequencing based on the result of ***,we also found 2 kinds of unreported SNPs,IVS-I-13G>A,IVS-II-310T>*** the tested samples,the frequency of 4 kinds SNP was high,among which 3 kinds SNPs—rs713040,rsl0768683,rsl609812,were carried *** *** gene diagnosis,in α-thal cases,the form of αα/--SEA accounted the major and CD41-42,IVS-Ⅱ-654,-28,CD17 are the main type of p-thal *** frequency of P-thal heterozygotes,homozygotes,and a-thal combining P-thal is not low,especially a-thal combining P-thal easily lead to misdiagnosis in clinical tests,which is significant for genetic counseling and accurate prenatal *** combining the whole P-globin gene sequencing can effectively detect the common P-thal mutation and even new

关键词： Thalassemia Genetic Testing Polymorphism Single Nucleotide Globins

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Genetic Diagnosis and Prenatal Diagnosis of Haemophilia B in China by DHPLC and Direct Sequencing

Genetic Diagnosis and Prenatal Diagnosis of Haemophilia B in...

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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： X Xiao XY.Lin SQ.Chen HY.Li YB.Guo WY.Jiang Department of Medical Genetics Medical SchoolSun Yat-Sen University

Introduction Denaturing High Performance Liquid Chromatography(DHPLC)is accurate,sensitive,economic and high-throughput for detecting *** aim to establish a feasible protocol to provide genetic diagnosis and prenatal diagnosis for Chinese hemophilia B patients and carriers by DHPLC and *** The genetic diagnosis was performed on 5 unrelated families,and 4 of them asked for prenatal *** The results of DHPLC and Sequence analysis indicated that the patients and carriers from five families have the pathogenic mutations,c.l022G>A(p.R341Q),c.484C>T(p.R162X),***35C>T(p.R379X),c.799C>T(p.H267Y)and c.l232G>T(p.S4III),*** c.484C>T(p.R162X),the other4 mutations were firstly reported firstly in Chinese *** prenatal diagnosis,one of the fetuses was found to be hemizygote with the c.484C>T(p.R162X) mutation and another two fetus were found to be carriers of c.484C>T/C(p.R162X/C) and c.799C>T/C(p.H267Y/H) *** remaining two fetuses were diagnosed as normal and it was verified after they were *** A combination of DNA direct-sequencing with DHPLC is rapid and accurate for the genetic diagnosis including prenatal diagnosis of hemophilia B ***,the level of F9 clotting activity in umbilical cord blood couldn't be used as a reference resource in the prenatal diagnosis of HB.

关键词： Hemophilia B Genetic diagnosis Prenatal diagnosis Denaturing High Performance Liquid Chromatography(DHPLC) Direct-sequencing

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Noninvasive prenatal diagnosis of common aneuploidies by semiconductor sequencing

Noninvasive prenatal diagnosis of common aneuploidies by sem...

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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： Liao C Yin AH Peng CF Fu F Yang JX Li R Chen YY Luo DH Zhang YL Ou YM Li J Wu J Mai MQ Hou R Wu F Luo H Li DZ Liu HL Zhang XZ Zhang K Prenatal Diagnosis Centre Guangdong Women and Children Hospital

Massively parallel sequencing(MPS) of cell-free fetal DNA from maternal plasma has revolutionized our ability to perform noninvasive prenatal *** approach avoids the risk of fetal loss associated with more invasive diagnostic *** present study developed an effective method for noninvasive prenatal diagnosis of common chromosomal aneuploidies using a benchtop semiconductor sequencing platform(SSP),which relies on the MPS platform but offers advantages over existing noninvasive screening techniques.A total of 2,275 pregnant subjects was included in the study;of these,515 subjects who had full karyotyping results were used in a retrospective analysis,and 1,760 subjects without karyotyping were analyzed in a prospective *** the retrospective study,all 55 fetal trisomy 21 cases were identified using the SSP with a sensitivity and specificity of 99.94%and 99.46%,*** SSP also detected 16 trisomy 18 cases with 100%sensitivity and 99.24%specificity and 3 trisomy 13 cases with 100%sensitivity and 100%***,15 fetuses with sex chromosome aneuploidies(10 45,X,2 47,XYY,2 47,XXX,and 1 47,XXY) were *** the prospective study,nine fetuses with trisomy 21,three with trisomy 18,three with trisomy 13,and one with 45,X were *** our knowledge,this is the first large-scale clinical study to systematically identify chromosomal aneuploidies based on cell-free fetal DNA using the SSP and provides an effective strategy for large-scale noninvasive screening for chromosomal aneuploidies in a clinical setting.

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Development of a platform for breeding by design of CMS lines based on anSSSL library in rice(Oryza sativaL.)

Development of a platform for breeding by design of CMS line...

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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： Ziju Dai Qing Lu Xin Luan Jian Cai Haitao Zhu Ziqiang Liu Ruizhen Zeng Zemin Zhang Shaokui Wang Lingjie Zheng Jialin Li Guiquan Zhang State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources South China Agricultural University

Cytoplasmic male sterility(CMS) is an important foundation for heterosis utilization andexploited widely to hybrid seed production in ***,the traditional experiencedriven and labor-cost breeding style left little room for sustainable improvement ofhybrid *** of molecular biology and genomics provides a chance toenhance hybrid rice breeding efficiency through breeding by *** this study,aline H121 B with rj3 and rf4 genes from XieqingzaoB(XB) was developed in theHuajingxian74(HJX74) genetic *** H121 B as maintainer line,threeisonuclear alloplasmic CMS lines,W-H121 A,D-H121 A and Y-H121 A,with the wildabortive(WA),dwarf wild abortive(DA),and Yegong abortive(YA) types of CMScytoplasm,respectively,were *** H121 B maintainer line was thenimproved by a way of breeding by *** favorable alleles,gs3 for long grain,Wxlfor low amylose content,and OsMADSSO for early heading date,in the chromosomalsubstituted segments of single segment substitution lines(SSSLs) were selected fromthe SSSL library in the HJX74 genetic background,and were pyramided together inH121 B through marker-assisted selection(MAS).The new maintainer line,H131 B,showing the phenotypes of longer in grain length,lower in amylose content,and earlierin heading date than those in H121 B,achieved the breeding goal ***131 B as maintainer line,three new isonuclear alloplasmic CMS lines,W-H131 A,DH131A and Y-H131 A,were *** results indicate that breeding by designin rice can be conducted under the platform of the HJX74 SSSL *** the platform for breeding by design of CMS lines will be helpful for sustainableimprovement of CMS lines in hybrid rice development.

关键词： Single segment substitution line Cytoplasmic male sterility Breeding by design Rice

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Genetic and cytological analysis of a novel type of low temperature-dependent intrasubspecific hybrid weakness in rice

Genetic and cytological analysis of a novel type of low temp...

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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： Chong-Yun Fu Feng Wang Bing-Rui Sun Wu-Ge Liu Jin-Hua Li Ru-Fang Deng Di-Lin Liu Zhen-Rong Liu Man-Shan Zhu Yi-Long Liao Rice Research Institute Guangdong Academy of Agricultural Sciences Guangdong Provincial Key Laboratory of New Technology in Rice Breeding Public laboratory South China Botanical GardenChinese Academy of Sciences

Hybrid weakness（HW） is an important postzygotic isolation which occurs in both intra- and inter-specific *** this study,we described a novel low temperature-dependent intrasubspecific hybrid weakness in the Fi plants derived from the cross between two indica rice varieties Taifeng A and *** plants showed growth retardation,reduced panicle number and pale green leaves with chlorotic *** assay showed that there were reduced cell numbers,larger intercellular spaces,thicker cell walls,and abnormal development of chloroplast and mitochondria in the mature leaves from HW Fi plants in comparison with that from both of the parental *** analysis revealed that HW was controlled by two complementary dominant genes Hw3 from VI134 and Hw4 from Taifeng ***3 was mapped in a 136 kb interval between the markers Indel 1118 and Indellll7 on chromosome 11,and Hw4 was mapped in the region of about 15 cM between RM182 and RM505 on chromosome 7,***-PCR analysis revealed that only LOCs11g44310,encoding a putative calmodulin-binding protein（OsCaMBP）,differentially expressed among Taifeng A,V1134 and their HW *** recombinant was detected using the markers designed based on the sequence of LOCs11g44310 in the BCiF2（Taifeng A//Taifeng A/V1134）***,LOCs11g44310 was probably the candidate gene of *** amplification suggested that LOCs11g44310 was present in VI134 and absent in Taifeng *** results of both parents' genomes further indicated that LOCs11g44310 was absent in Taifeng *** search revealed that LOCs11g44310 had one copy in the japonica genomic sequence of Nipponbare,and no homologous sequence in the indica reference sequence of *** results indicate that Hw3 is a novel gene for inducing hybrid weakness in ***,based on the sequencing results of both parents' genomes,some polymorphic InDel markers were developed,and Hw4 was further narrowed down in an about 2 cM interval and three polymorphic InDe

关键词： LOC Genetic and cytological analysis of a novel type of low temperature-dependent intrasubspecific hybrid weakness in rice

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薏苡丙酮酸脱羧酶基因克隆及淹水表达响应分析

薏苡丙酮酸脱羧酶基因克隆及淹水表达响应分析

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者：陈大清陈双建何建君李亚男仲恺农业工程学院生命科学学院长江大学农学院

薏苡（Coix lacryma/obi L.）为禾本科C植物,起源于东南亚的热带和亚热带地区,是古老的粮药兼用作物,具有较高的营养和医学价值,在中国至少有6000-10000年的栽培历史,同时具有耐涝、耐盐、耐早等多抗性。我们以薏苡为材料,克隆丙酮酸脱... 详细信息

薏苡（Coix lacryma/obi L.）为禾本科C植物,起源于东南亚的热带和亚热带地区,是古老的粮药兼用作物,具有较高的营养和医学价值,在中国至少有6000-10000年的栽培历史,同时具有耐涝、耐盐、耐早等多抗性。我们以薏苡为材料,克隆丙酮酸脱羧酶基因（pyruvatedecarboxylasel,PDC1）序列,应用RT-PCR分析基因对淹水表达的响应。根据玉米、水稻PDC1蛋白保守序列和甘蔗PDC片段设计两对简并引物,分别从薏苡cDNA中克隆出长度为471bp和972bp的两个片段;运用RACE技术克隆了薏苡PDC1 3'端编码序列,测序后对序列进行整合,通过分析重叠区的摇摆碱基密码子使用的偏好性（http://***/codord）,确定蛋白序列编码。整合后的核酸序列序列长度为1398bp,与甘蔗PDC片段和玉米PDC3相似度为97%,与玉米和籼稻PDC2相似度分别为95%和91%;蛋白质序列与玉米PDC3相似度最高为97%,与籼稻和粳稻PDC2蛋白相似度为96%。RACE5'端上游序列克隆,获得了152bp片段,在线比对发现从薏苡中克隆的序列与玉米（Zay mays）、水稻（Oryza sativajaponicaGroup）的PDC基因家族的基因序列最大相似度均超过88%以上（http://***/***）。蛋白序列在线比对BlastP（http://***/cgi-bin/dnaa）结果显示:与大麦、玉米PDC3和水稻PDC2相似度最高,分别为98%,96%和96%,与玉米和水稻其它PDC蛋白相似度也在80%以上。将克隆薏苡PDCI 5'端编码序列的部分与前述片段整合成1507bp序列。用ClustalX软件将薏苡PDC1序列及其蛋白质序列与水稻、玉米、甘蔗、稗和拟南芥PDC家族共15个基因的核酸和蛋白序列分别进行比对,并采用邻接法（NJ）,运用Mega4.0软件构建了进化树,初步确认我们克隆的薏苡PDC1与甘蔗PDC片段、玉米PDC3三者间都有很高的亲缘性,暗示我们初步获得了薏苡PDC1序列,进一步将通过两端特异引物,扩增全长cDNA序列进行验证。此外,我们采用半定量RT-PCR技术,以18sRNA为内参,初步研究了薏苡根和叶在淹水胁迫下的表达特点。结果表明:薏苡经淹水处理后,根和叶中PDC1的表达量在0-24h呈上升趋势,并达到最高值,24h后下降。

关键词：薏苡丙酮酸脱羧酶基因克隆淹水表达

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