OBJECTIVETo explore the association between the short tandem repeat polymorphism of exon 5 of MICA gene (MICA-STR) and nasopharyngeal carcinoma (NPC) in a southern Chinese population.

METHODSOne hundred and twenty-seven consecutive NPC patients and 112 randomly selected normal controls residing in southern China mainland were analyzed for MICA-STR allelic variation and MICA gene deletion by fluorescent polymerase chain reaction-gene scanning and polymerase chain reaction-sequence specific priming.

RESULTSMICA*A9 was observed at significantly higher frequency in the NPC patient group than in the control group (relative risk = 2.524, P = 0.001,Pc = 0.006); whereas MICA*A5.1 was present at significantly lower frequency in the NPC patient group than in the control group (RR = 0.418, P = 0.0004, Pc = 0.0026). Further analysis revealed that MICA*A9 was over-represented in male NPC patients, compared with male controls (RR = 3.23, P = 0.00095, Pc = 0.006); whereas MICA*A5.1 was present at significantly lower frequency in male NPC patients, compared with male controls (RR = 0.372, P = 0.0007, Pc = 0.004). None of the MICA-STR variants showed statistically significant frequency difference between female NPC patients and female controls (Pc > 0.05).

CONCLUSIONMICA-STR polymorphism is associated with NPC, and MICA*A9 is a genetic susceptibility marker of male individuals for NPC in a southern Chinese population.

Adult ; Asian Continental Ancestry Group ; genetics ; China ; Exons ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Histocompatibility Antigens Class I ; genetics ; Humans ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Nasopharyngeal Neoplasms ; ethnology ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics

OBJECTIVETo examine the relationship between intercellular adhesion molecule-1 (ICAM-1) gene polymorphism and ischemic stroke (IS) in Chinese Zhuang populations.

METHODSThe K469E polymorphism in the exon 6 of ICAM-1 gene was detected by polymerase chain reaction-restriction fragment length polymorphism analysis and DNA sequencing in 205 patients with IS of Zhuang nationality and in 210 healthy controls, and the serum level of ICAM-1 was determined by enzyme-linked immunosorbent assay.

RESULTSThe IS group showed significantly higher serum levels of ICAM-1 than did the control group (P < 0.01). There was significant difference in frequencies of allele and genotype in K469E polymorphism between IS and control groups, respectively (P < 0.05). The K allele carriers had 1.424 times the risk of suffering from IS as compared with the E allele carriers (OR = 1.424, 95% CI: 1.071 - 1.894); the serum ICAM-1 level of E allele carriers was significantly higher than that of K allele carriers (501.24 +/- 139.56 ng/ml vs 475.17 +/- 118.35 ng/ml, P < 0.01).

CONCLUSIONThere is an association between ICAM-1 gene K469E polymorphism and IS, and E allele may be a genetic risk factor of IS among Guangxi Zhuangs, in which the ICAM-1 E allele carriers may have up-regulated expression of ICAM-1 and hence are at a higher risk of ischemic stroke.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Brain Ischemia ; ethnology ; genetics ; China ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA

OBJECTIVETo determine a complex chromosomal rearrangement by advanced molecular cytogenetic techniques and analyze its clinical effect.

METHODSA complex chromosomal rearrangement (CCR) involved in chromosomes 5, 16 and 20 in a 29-year-old male carrier was determined by chromosomal microdissection and multicolor fluorescence in situ hybridization (M-FISH), and family degree investigation was further performed.

RESULTSThe karyotype of the case was a complex chromosomal translocation among chromosomes 5, 20 and 16, and accompanied with a band of chromosome 20 inserted into chromosome 5. His mother and sister both had the same abnormal karyotype by familial investigation.

CONCLUSIONThe combined use of M-FISH and chromosome microdissection is a powerful tool to determine CCR. The complex chromosomal rearrangement could be transmitted stably in the family, but still the carriers could give birth to a healthy baby by chance.

Adult ; Chromosomes, Human, Pair 10 ; genetics ; Chromosomes, Human, Pair 16 ; genetics ; Chromosomes, Human, Pair 20 ; genetics ; Chromosomes, Human, Pair 5 ; genetics ; Cytogenetic Analysis ; methods ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Pregnancy ; Translocation, Genetic

OBJECTIVETo study the association between cholesteryl ester transfer protein (CETP) gene polymorphism and insulin resistance in type 2 diabetes.

METHODSCETP-TaqIB gene was genotyped with polymerase chain reaction-restriction enzyme fragment polymorphism analysis in 108 patients with type 2 diabetes and in 60 normal controls. Plasma lipids, fasting insulin, insulin sensitivity index and insulin resistance index were determined in 108 patients with type 2 diabetes.

RESULTSThe distribution of CETP-TaqIB genotypes and B1B2 allele frequency in the patients with type 2 diabetes were similar to that in general population. High density lipoprotein cholesterol (HDL-C), apolipoprotein A1(apoA1) and insulin sensitivity index (ISI) levels were significantly higher in B2B2 genotype than in B1B1 genotype. Fasting insulin (FINS) and Homeostasis model assessment-insulin resistance (HOMA-IR) levels were significantly lower in B2B2 genotype than in B1B1 genotype. No significant differences in triglyceride (TG), total cholesterol(TC),low density lipoprotein cholesterol (LDL-C), apolipoprotein B (apoB) levels were observed among different CETP-TaqIB genotype groups. By multivariate analysis, the determinants of ISI and HOMA-IR were body mass index (BMI), TC, HDL-C and CETP-TaqIB genotype.

CONCLUSIONThe CETP-TaqIB genotype is independently associated with insulin resistance and lipid metabolism. It may be an important risk factor of insulin resistance in type 2 diabetes.

Adult ; Aged ; Aged, 80 and over ; Cholesterol Ester Transfer Proteins ; genetics ; metabolism ; Diabetes Mellitus, Type 2 ; genetics ; metabolism ; Female ; Gene Frequency ; Genotype ; Humans ; Insulin Resistance ; Lipid Metabolism ; Male ; Middle Aged ; Polymorphism, Genetic ; genetics

OBJECTIVETo establish the restriction fragment differential display-polymerase chain reaction (RFDD-PCR) as an efficient technique for constructing and studying the gene expression profile of human tissues.

METHODSThe tissues of mamma adenocarcinoma (T), cancerometastasis lymph node (L) and normal mammary (N) from one mammary infiltrating ductal carcinoma case were collected, and the gene expression profile of each kind of tissue was constructed using RFDD-PCR technique at equal pace according to the operating manual of Qbio-gene Company. Then all fragments of the three gene expression profiles were separated and displayed by electrophoresis. With the use of gene database at the website http://www.Qbio-gene.com/display, the authors identified the names of the probable fragments by bioinformatics analysis. Through comparison of the three profiles, the numbers and types of most differentially expressed gene fragments were displayed.

RESULTSThe expression profiles of the three kinds of tissue have been constructed covering 1716 fragments of mammary adenocarcinoma, 1769 of cancerometastasis lymph nodes and 1922 of normal mammary tissue. Among these 5407 fragments, 39.39% were exactly the same. While 33.9% sequences of T and L showed differences in abundance or presence, 40.9% of T and N and 39.6% fragments of L and N were observed differentially expressed. These differentially expressed gene fragments were found to relate with metastasis, differentiation, inflammation and so on.

CONCLUSIONRFDD-PCR is an efficient technique for research in human diseases genomics as a mass screening for complete gene expression profile with high-flux. Through comparison among three or more profiles, the screening for candidate genes of a certain disease can be accomplished, and there is probably a chance to identify novel gene or expressed sequence tag.

Adenocarcinoma ; genetics ; Breast Neoplasms ; genetics ; Carcinoma, Ductal, Breast ; genetics ; Computational Biology ; Electrophoresis ; methods ; Female ; Gene Expression Profiling ; methods ; Humans ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA ; methods

OBJECTIVETo introduce the application of denaturing high-performance liquid chromatography (DHPLC) in the diagnosis of childhood type spinal muscular atrophy (SMA).

METHODSExon 7 and flanking area of survival motor neuron (SMN) gene were amplified by PCR in 1 standard sample, 25 normal individuals and 25 patients with SMA. The PCR products were then directly loaded onto the DHPLC system after denaturing and annealing. Different DNA segments were separated by changing the concentration of buffer A relative to that of buffer B.

RESULTSDifferent DNA segments were separable on the DHPLC chromatogram. Three peaks including SMN1/SMN2 heteroduplex peak, SMN2 homoduplex peak and SMN1 homoduplex peak were detected in 23 out of 25 normal individuals. Only SMN1 homoduplex peak was detected in 2 normal individuals and the standard sample, indicating the deletion of SMN2 On the contrary, only the SMN2 homoduplex peak was detected in 22 out of 25 patients with SMA, indicating deletion of SMN1. The three peaks as those of normal individuals were detected in the other 3 patients, indicating no SMN1 or SMN2 deletion.

CONCLUSIONAs a new technology for diagnosing SMA, DHPLC is sensitive, accurate, rapid and convenient.

Chromatography, High Pressure Liquid ; methods ; Exons ; genetics ; Humans ; Muscular Atrophy, Spinal ; diagnosis ; genetics ; Polymerase Chain Reaction ; Reproducibility of Results ; SMN Complex Proteins ; genetics ; Sensitivity and Specificity ; Survival of Motor Neuron 1 Protein ; genetics ; Survival of Motor Neuron 2 Protein

OBJECTIVETo investigate the molecular genetics basis for a novel HLA allele, HLA-B*5614, in Chinese population.

METHODSDNA was extracted from whole blood by salting-out method. The HLA-B exons 2-4 of the proband was amplified and the amplified product was cloned using TOPO cloning sequencing kit to split the two alleles apart. Both strands of exons 2,3 and 4 of chosen colonies were sequencing. The PCR-SSP was performed to confirm the mutations detected by sequencing.

RESULTSThe sequencing results showed the HLA-B alleles of the proband as B*1502 and the novel allele. The sequences of the novel allele have been submitted to GenBank (AY601726, AY610727, AY610728). After BLAST analysis, the novel allele differs from B*5608 by a single nucleotide at position 277G-->C in exon 2. This results in an amino acid change from Gly to Arg at codon 93.

CONCLUSIONThis allele is a novel allele and has been officially named B*5614 by the WHO Nomenclature Committee.

Alleles ; Exons ; genetics ; HLA-B Antigens ; genetics ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA ; methods

OBJECTIVETo search the interaction between reduced folate carrier gene (RFC1 A80G) polymorphism of children with neural tube defects (NTDs) and maternal periconceptional no supplementation of folic acid. The purpose is to provide the epidemiological evidence for finding genetic marker of NTDs.

METHODSRFC1 (A80G) genotype was detected using PCR-restricted fragment length polymorphism for the blood DNA of 104 trios with NTDs-affected child, and 100 control families with non-malformed control children. The authors investigated the gene-environment interactions between the offspring RFC1 genotype and maternal periconceptional folic acid supplementation through a case-control study.

RESULTSIt was observed that the offspring with the GG genotype were associated with a 2.56-fold increased risk of NTDs when compared to those with the AA genotype (OR = 2.56; 95% CI = 1.04-6.36) in this population under investigation. The risk of mothers who did not take folic acid for having an NTDs-affected infants was 7.69 (95% CI = 2.86-21.75). Among the mothers who did not utilize folic acid supplements, the NTDs risk was 3.30 (95% CI = 1.15-9.65) for offspring with the GG genotype, compared to the reference (AA) genotype. Children who had the GG genotype and whose mothers did not take folic acid had an elevated risk for NTDs (OR = 8.80, 95% CI = 2.86 - 29.82), compared to "offspring with AA or GA genotype" and "maternal folic acid use", the interactive coefficient being 1.45.

CONCLUSIONThe above findings indicate that the RFC1 genotype (GG) is a possible susceptible gene marker for an increased NTDs risk in Chinese population, and there is a potential gene-nutrient interaction between offspring RFC1 GG genotype and maternal periconceptional intake of folic acid on the risk of NTDs. However,the sample size of this study was limited, a larger sample of population-based study is required to pursue the initial observation.

Adult ; Case-Control Studies ; Child ; Child, Preschool ; Dietary Supplements ; Female ; Folic Acid ; administration & dosage ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Membrane Transport Proteins ; genetics ; Neural Tube Defects ; genetics ; prevention & control ; Polymorphism, Genetic ; genetics ; Reduced Folate Carrier Protein ; Vitamin B Complex ; administration & dosage

OBJECTIVETo investigate the relationship between apolipoprotein A5(apoA5) - 1131T > C polymorphism and the susceptibility of coronary artery disease (CAD) in Chinese.

METHODSThe restriction fragment length polymorphism of apoA5 gene - 1131T > C was studied using PCR in a case-control study which enrolled 235 patients with CAD diagnosed by angiography and 262 healthy controls from Jiangsu province.

RESULTSThe frequencies of T, C allele were 59.57%ì40.43% and 65.65%, 34.35% in CAD group and control group respectively. There was statistically significant difference in allele frequencies between CAD group and control group (P < 0.05). The susceptibility to CAD for the CC genotype was much higher than that for wild type TT (OR = 1.872, 95% CI = 1.039 - 3.376, P = 0.037), even after the use of Logistic regression models (OR = 2.285, 95% CI = 1.222 - 4.274, P = 0.012). In control group, there was significant difference in TG levels among different genotypes, the C allele carriers had higher serum TG concentration (P = 0.007).

CONCLUSIONapoA5 - 1131T > C polymorphism is associated with an increased risk of CAD and is also in strong association with serum TG levels.

Aged ; Apolipoprotein A-V ; Apolipoproteins A ; genetics ; Asian Continental Ancestry Group ; genetics ; China ; Coronary Artery Disease ; blood ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Humans ; Lipids ; blood ; Logistic Models ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Triglycerides ; blood

OBJECTIVETo study the clinical features and to identify homeobox D13 (HOXD13) gene mutation of the affected individuals in a Chinese synpolydactyly (SPD) kindred.

METHODSClinical data and peripheral blood samples of SPD family members were obtained through field investigation. For every member of this pedigreeìthe fragment containing mutational hot spots of HOXD13 was amplified by PCR for mutation screening. To examine whether there is any other mutation within coding sequence of HOXD13, exon 1 and exon 2 of HOXD13 were also amplified by PCR. All the amplified fragments were electrophoresed on 2% agarose gels and then the mutant fragments were electrophoresed on 5% polyacrylamide gels to be separated. Purified PCR products of normal and selected mutant alleles were directly sequenced.

RESULTSComparing the HOXD13 coding sequence of the affected individuals with HOXD13 sequence in the GenBank and with that of the unaffected, an inserted segment coding 8 alanine residues within HOXD13 was found segregating with the disorder. This mutation is also termed polyalanine expansion. The 8-alanine expansion can be interpreted as a reduplication of normal alanines 5-12.

CONCLUSIONThe results suggest that synpolydactyly in this kindred may be caused by polyalanine expansion in HOXD13.

Base Sequence ; China ; DNA Mutational Analysis ; Female ; Homeodomain Proteins ; genetics ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Syndactyly ; genetics ; Transcription Factors ; genetics