Detection of HOXA3 gene expression and apoptosis in myocardial tissues of children with tetralogy of fallot / 中华实用儿科临床杂志

Objective To explore the potential role of HOXA3 gene in children with tetralogy of fallot (TOF) by detection the expression of HOXA3 mRNA and protein levels,as well as the apoptotic cardiomyocytes in the developing heart tissues.Methods Twenty-two surgical samples from sporadic cases of TOF determined by prenatal color Doppler ultrasound and autopsy [gestational age(25.67 ± 7.68) weeks,TOF group] were examined with quantitative real-time PCR and Western blot to evaluate the expression of HOXA3 gene.Twelve age-matched autopsies without heart structural abnormalities [gestational age (26.55 ± 6.36) weeks,control group] were also included.Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL) assay was performed to clarify the apoptosis of cardiomyocytes.The difference between the 2 groups and the correlation analysis of HOXA3 level and the apoptotic cardiomyocytes were performed with SPSS 13.0 software.Results Compared with control group,HOXA3 mRNA expression of the outflow tract of the right ventricle from the TOF group was significantly reduced(P ＜ O.01).Western blot also confirmed that the HOXA3 protein was accordingly reduced(P ＜0.01).The proportion of apoptotic cardiomyocytes in samples of TOF group was significantly greater than that of control group (P ＜ 0.01).The proportion of apoptotic cells was strongly correlated with the mRNA and protein expression of HOXA3 (r =-0.566,-0.759,all P ＜ 0.01).Conclusions Reduction of HOXA3 gene expression and the increase of apoptotic cardiomyocytes at the crucial stage during heart development may play a potential role in the onset of TOF.

Association of the GLI gene with ventricular septal defect after the susceptibility gene being narrowed to 3.56 cM in 12q13 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Our previous research has suggested that genes around D12S1056 in 12q13 may confer susceptibility to ventricular septal defect (VSD) in humans. The present study was to define the chromosome region assignment by transmission disequilibrium test (TDT), and to identify the important candidate gene by family-based association study and haplotype analysis.</p><p><b>METHODS</b>Surrounding D12S1056, ten microsatellite markers including D12S329, D12S305, D12S1662, D12S1056, D12S1293, D12S334, D12S102, D12S83, D12S1655 and D12S1691 were chosen, and TDT was performed in 62 nuclear family trios each consisting of an affected child and two healty parents. Subsequently, the GLI gene, a positional candidate gene that maps to the target region, was selected for further analysis. Three single nucleotide polymorphisms (SNPs), G11888C, G11388A, and G11625T, were selected for family-based association study and haplotype analysis.</p><p><b>RESULTS</b>VSD was significantly associated with all selected markers except D12S1691 [72.2 centi morgen (cM)] and D12S1700 (75.76 cM). VSD was also significantly associated with G11888C (chi(2) = 5.918, P = 0.015), G11388A (chi(2) = 8.067, P = 0.005), and G11625T (chi(2) = 11.842, P = 0.001). Haplotype analysis showed a strong linkage disequilibrium between G11888C and G11388A (D' = 0.999), but in significant (chi(2) = 1.035, df = 2, P > 0.05).</p><p><b>CONCLUSIONS</b>The susceptibility gene of VSD was mapped to 3.56 cM in 12q13 by TDT, and the GLI gene, an important candidate in the target region, was associated with VSD.</p>

Analysis of single nucleotide polymorphisms and haplotypes in HOXC gene cluster within susceptible region 12q13 of simple congenital heart disease / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>In the candidate region 12q13 of simple congenital heart disease(CHD), four single nucleotide polymorphisms(SNPs) in HOXC4 gene were chosen in order to investigate the distribution of SNP and haplotypes in simple CHD patients and normal people.</p><p><b>METHODS</b>The genotype of 4 SNPs in 108 simple CHD patients and 200 normal people were analyzed by restriction fragment length polymorphism(RFLP) and denaturing high-performance liquid chromatography(DHPLC). The statistical contingency table method was used to analyze SNP genotype frequency and gene frequency in patients and control group; then, the haplotypes were established and their frequencies in the two groups were assessed by PHASE software.</p><p><b>RESULTS</b>C16476T polymorphism was not detected; A17860G located in 3' flanking sequence of HOXC5 gene displayed significant difference between the two groups. The G allele frequency in simple CHD patients was higher than that in healthy controls(P < 0.05); the distribution of frequencies of 4 haplotypes showed significant difference(P < 0.01).</p><p><b>CONCLUSION</b>The A17860G located in 3'flanking sequence of HOXC5 gene is associated with simple CHD; the risk of CHD in the persons with G17860 is higher than that in those with A17860. the haplotype of 3 SNPs may be linked with the susceptible gene of simple CHD.</p>

Analysis and application of SCA1 and SCA3/MJD gene CAG repeats in Han population in Northeastern China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the normal range of (CAG)n in spinocerebellar ataxia type 1 (SCA1) gene and spinocerebellar ataxia type 3 (SCA3/MJD) gene in 110 normal subjects of Han population in Northeastern China, to assess the genotypes for clinically diagnosed spinocerebellar ataxia(SCA) individuals including 25 patients from 8 families and 6 sporadic patients, and to make presymptomatic and prenatal diagnosis.</p><p><b>METHODS</b>DNA fragments from the normal subjects and the patients were detected by fluorescence-PCR. Homozygosities were selected for DNA sequencing.</p><p><b>RESULTS</b>The normal ranges of (CAG)n of SCA1 and SCA3/MJD were 20-39 and 14-38 repeats respectively, SCA1 was found mostly to be 26 and 27 repeats, allele frequency 34.09% and 20.91%; heterozygosity was 84.55%, SCA3/MJD was found mostly to be 14 repeats, allele frequency 39.55%, heterozygosity was 78.18%.(CAG)(68) of SCA3/MJD gene of one affected individual had been found in a family but no CAG mutative expansion in related members was observed.</p><p><b>CONCLUSION</b>The normal ranges of CAG repeats vary with areas and races. SCAs genotyping is the first choice in presymptomatic and prenatal diagnosis.</p>

Analysis on genetic polymorphism of 14 short tandem repeat loci on chromosome 7p14-15 and 12q13 in Chinese north Hans / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the genetic polymorphism of 6 short tandem repeat (STR) loci on chromosome 7p14-15 and 8 STR loci on chromosome 12q13 in Chinese north Hans.</p><p><b>METHODS</b>Fluorescence-labeling polymerase chain reaction and capillary electrophoresis were used to analyze the genetic polymorphism of 100 randomly selected individuals from Chinese north Han nationality at 6 STR loci (D7S1808, D7S2250, D7S2251, D7S683, D7S656 and D7S528) on chromosome 7p14-15 and 8 STR loci(D12S1056, D12S1293, D12S83, D12S1655, D12S1662, D12S334, D12S137 and D12S102) on chromosome 12q13.</p><p><b>RESULTS</b>In the Chinese north Han population, 7 alleles and 24 genotypes, 8 alleles and 27 genotypes, 7 alleles and 22 genotypes, 4 alleles and 10 genotypes, 6 alleles and 17 genotypes, 5 alleles and 13 genotypes were observed at D7S1808, D7S2250, D7S2251, D7S683, D7S656 and D7S528. The heterozygosities at the above 6 STR loci were 86%, 88%, 83%, 79%, 85% and 80%, respectively. Five alleles and 15 genotypes, 5 alleles and 15 genotypes, 8 alleles and 29 genotypes, 6 alleles and 17 genotypes, 6 alleles and 17 genotypes, 6 alleles and 19 genotypes, 5 alleles and 13 genotypes, 7 alleles and 24 genotypes were observed at D12S1056, D12S1293, D12S83, D12S1655, D12S1662, D12S334, D12S137 and D12S102. The heterozygosities at the above 8 STR loci were 86%, 84%, 87%, 82%, 84%, 85%, 81% and 89%, respectively.</p><p><b>CONCLUSION</b>The distributions of allele frequencies of 6 STR loci on chromosome 7p14-15 and of 8 STR loci on chromosome 12q13 were consistent with the Hardy-Weinberg equilibrium. The highly genetic polymorphism was observed in Chinese north Han population.</p>