Objective To investigate the distribution of single nucleotide polymorphisms(SNPs) on retinol binding protein 4(RBP4) genes and forkhead box O1 (FOXO1) gene, and their relationships with the occurrence of type Ⅱ diabetes mellitus (T2DM) in Chinese Han population. Methods Totally ten SNPs on RBP4 and FOXO1 were determined in 384 T2DM patients and 384 normal controls by TaqMan probe genotyping and agarose gel electrophoresis methods. And their serum level of fasting blood glucose (FBG), total cholesterol (TC) and trigly- ceride (TG) were also estimated. Results For RBP4, there was no significance for various genetypes and alleles including - 803 G > A, + 5169 C > T, and + 6969 G > C between two groups (P > 0.05). Each genotype had no relationships with T2DM (using adjusted logistic regression models). No haplotype was associated with T2DM. For FOXO1, among seven SNPs typed, significant variation was found in the frequency distribution of rs7324943 G/T in the two groups(χ~2=4.02, P = 0.044), and further stratification analysis showed that in subjects of aged 40 and non-hypertension, there was a higher risk of T2DM in GT heterozygous carriers than in GG homozygous carriers (OR = 1.47, 1.80), T allele carriers showed higher risk than non-T carriers (OR = 1.42,1.79). For rs17592236 C/T, though no significant frequency variation was found between two groups (χ~2 = 0.39, P = 0.401), but in subjects of aged ≤ 40, stratification analysis showed dramatically increased risk of T2DM in CT and TT carriers than in CC carriers (OR = 6.33,10.15), T allele carriers showed 7. 11-fold higher risk than non-T carriers. A haplotype CT related to T2DM susceptibility was also found, which could decrease the risk of its carriers by 28%. Conclusions For BBP4, the polymorphisms of - 803 G > A, + 5169 C > T, and + 6969 G > C had no relationships with T2DM in Chinese Han population. For FOXO1, the polymorphism of rs7324943 G/T,rs17592236 C/T and a haplotype CT were found related to the susceptibility of T2DM in Chinese Han population. Yet further studies are necessary to explain the impact of these polymorphisms on the disease occurrence.

To describe the fractal feature of CGR (Chaos-game representation) graph of genomes sequences, a multifractal theory is presented in the analysis. By studying the effect of three probability sets on the scale invariance range, the probability set with the best scale invariance is chosen, and then the smooth general dimension spectrum and multifractal spectrum are calculated. The experimental result shows that the probability set composed of the relative probability has the best scale-invariance performance. The scale invariance has three different variance regions, which indicate that genomes sequence segments with different lengths have different distribution rules. It is concluded that the multifractal method is effective for describing the fractal feature of CGR graph of genomes sequences.
Algorithms ; Base Sequence ; Computer Simulation ; Game Theory ; Genome ; Humans ; Mathematical Computing ; Sequence Analysis, DNA ; statistics & numerical data

Biosequence analysis is the primary research field of bioinformatics. In this field, useful information can be extracted by comparison analysis methods. Among them, sequence alignment is the most common comparison method. However the sequence comparison by alignment, which assumes conservation of contiguity between homologous segments, is at odds with genetic recombination. Especially for the multisequence alignment, there exists the difficulty in the complexity of calculation. Therefore, alignment-free sequence comparison methods are required. In this paper, two main categories of alignment-free sequence comparison methods are reviewed. The first one is based on the word (oligomer) frequency and its distribution. The sequences are compared using the distances defined in a Cartesian space by the frequency vectors. In the second category, sequences are compared using Kolmogorov complexity and chaos theory.
Algorithms ; Computational Biology ; Sequence Alignment ; Sequence Analysis ; methods

Objective: To clone human interleukin 10(hIL-10) gene full-length cDNA sequence and to construct its eukaryotic expression vector in Chinese hamster ovary(CHO) cells. Methods: cDNA fragment encoding hIL-10 gene was amplified from human peripheral normal white blood cells by RT-PCR and confirmed by DNA sequencing.The ds-DNA was inserted into pcDNA3 vector.This recombinant expressing vector was transfected into CHO cell.The IL-10 molecules expressed were detected by ELISA and the role of inhibition of IL-10 was determined with MTT. Results: The cDNA encoding hIL-10 was cloned by RT-PCR and inserted into T-easy vector and sequence analysis verified that the cloned fragment was hIL-10 cDNA.The IL-10 was expressed in the CHO and it blocked the lymphocyte transformation.Conclusion: The eukaryotic expression plasmid was constructed successfully,which will contribute to further studies on the role of hIL-10 in autoimmunity,transplantation immunity and inflammatory disease.

OBJECTIVETo detect the mutation sites of exons 2, 20, 11A and 11B in Chinese patients with breast cancer.

METHODSA total of 86 patients with breast cancer without blood relationship were randomly selected. Polymerase chain reaction (PCR) and double-strand DNA direct sequencing were applied.

RESULTSNo mutations, especially deletions were found in exons 2, 20 and 11 with carefully checking the sequencing results, although they were reported frequently in Europe populations with breast cancer. We found one polymorphism in exon 11, with high frequency, and in the test of chi-square, the frequencies of two alleles had no significant difference between the patients and controls.

CONCLUSIONThe above results suggest this SNP may not be associated with the breast cancer in Chinese population, and indicates that the gene sequence of what we have studied doesn't account much for occurrence of the breast cancer in the population of China.

Asian Continental Ancestry Group ; genetics ; BRCA1 Protein ; genetics ; Breast Neoplasms ; genetics ; Exons ; Female ; Gene Frequency ; Humans ; Mutation ; Polymorphism, Genetic

OBJECTIVETo figure out the polymorphism of three Y-STR loci in isolated populations and explore the consanguinity of the populations with the use of Y-STR.

METHODSMale samples were selected from two isolated populations(80 and 60 males) in Zhejiang province and one open population (36 males), genescan was performed with males' DNA by genescan technology with ABI PRISM 377 sequencer at Y chromosome loci DYS388, DYS390 and DYS395.

RESULTSDYS388, DYS390, DYS395 allele counts in Yushan island population, Taohua island population and open population were 8, 9, 7, 5, 6, 7 and 6, 6, 5 respectively. Gene diversity was between 0.70-0.80 in the three populations. There was no difference in distribution of allele frequency and shared genotypes between the isolated populations and the open population by statistical test. Genetic distance is long between Taohua island population and open population, short between Yushan island population and open population, and moderate between Yushan island population and Taohua island population.

CONCLUSIONThe main allele is 129 at DYS388; 215 at DYS390; and 119 at DYS395. The distribution of allele frequency and gene diversity at DYS388, DYS390, DYS395 loci, and the shared genotypes between populations as well as the genetic distance are unable to explain the blood relationship between the isolated and open populations, suggesting the additional studies in large sample size will be necessary to use Y-STR for exploring the blood relationship between populations.

Alleles ; China ; Gene Frequency ; Genetics, Population ; Humans ; Male ; Microsatellite Repeats ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics ; Y Chromosome ; genetics

OBJECTIVETo investigate the association of two single nucleotide polymorphisms (SNPs) of beta 2-adrenoceptor (beta 2-AR) gene with hypertension in elderly patients.

METHODSThe study samples were collected from unrelated Chinese Han population of Dabie Mountain in Anhui province. Eighty-six elderly patients with hypertension and 43 controls were selected. Genotypes of +1053 and +1239 SNPs were typed by polymerase chain reaction-restriction fragment length polymorphism.

RESULTSThe frequencies of the two SNPs complied well with the Hardy-Weinberg equilibrium in normal group. The distribution of genotypes AA, GA,GG of the SNP at locus +1239 in moderate and severe hypertension group was significantly different from that in normal group (chi square=8.67, P<0.05). There were evident differences in the frequencies of alleles of the two groups (chi square=4.02, P<0.05). No significant difference was observed in the distribution of genotypes of the SNP at locus +1053 between the two groups.

CONCLUSIONThese data indicate that the SNP at locus +1239 of beta 2-AR gene is associated with hypertension in elderly patients.

Aged ; Aged, 80 and over ; Alleles ; DNA ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Receptors, Adrenergic, beta-2 ; genetics

Objective　To study whether human umbilical cord blood CD34+ cells transduced with human aldehyde dehydrogenase class-1 (ALDH-1) and multidrug resistance gene (MDR1) have increases resistance to 4-Hydroperoxycyclo-phosphamide (4-HC) and P-glycoprotein effluxed drugs. Methods　A bicistronic retroviral vector G1Na-ALDH1-IRES-MDR1 was constructed and used to transfect the packaging cell lines GP+E86 and PA317 by LipofectAMINE method, using the medium containing VCR and 4-HC agents for cloning selection and ping-ponging supernatant infection between the ecotropic producer clone and the amphotropic producer clone, we obtained high titer amphotropic PA317 producing cells with high titers up to 5.6×105 CFU/ml. Cord blood CD34+ cells were transfected repeatedly with supernatant of retrovirus containing human ALDH-1 and MDR1cDNA under the stimulation of hemopoietic growth factors. Results　Bicistronic retroviral vector construction was verified by restriction endonuclease analysis. Polymerase chain reaction (PCR), reverse transcription (RT)-PCR, Southern blot, Northern blot, fluorescenceactivated cell sorting (FACS) method and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analyses showed that dual drug resistance genes have been integrated into the genomic DNA of cord blood CD34+ cells and expressed efficiently. The transgenes recipient cells confered 4-fold stronger resistance to 4-HC and 5.5 to 7.2-fold P-glycoprotein effluxed drug than untransduced cells. Conclusion　The bicistronic retroviral vector-mediated transfer of two different types of drug resistance genes into human cord blood CD34+ cells and co-expression provided an experimental foundation for improving combination chemotherapy tolerance in tumor clinical trial.

Objective　The study was conducted to investigate single nucleotide polymorphism(SNP) in beta2-adrenoceptor(β2-AR) gene and the distribution of these identified SNPs in Chinese Han ethnic group.Methods　β2-AR gene was sequenced to detect SNPs by fluorescent labeling automatic sequencing method in 80 unrelated samples from territory of Dabie Mountain in Anhui province.Results A total of 8 SNPs were identified in length of 3.8 kb, including 5 SNPs in code region, 3 SNPs in regulatory region. Although the variations, -468C to G, -367T to C, -47C to T,-20T to C,+79C to G,+100G to A,+491C to T,+1098T to C have been identified in other ethnic groups, they have not been found in our study. The allele distribution of SNPs is in good unity with the Hardy-Weinberg equilibrium.Conclusion　The distribution of SNPs in β2-AR gene is not equable and the SNPs in different ethnic groups differ greatly. The allele distribution of SNPs conforms well to the Hardy-Weinberg equilibrium.

Retroviral vectors are wildly used as vehicles for gene transfer into hematopoietic cells based on its potency for efficient gene delivery and integration of transgene in host genome. The development of better transduction protocols depends on gene markers that allow a rapid detection and effective selection of genetically transduced cells. In this study, the enhanced green fluorescent protein (EGFP), a gene that is optimized for detection and expression in mammalian cells, was firstly amplified and cloned by high-fidelity PCR. The vector LGSN carrying EGFP gene was then constructed and the retroviral producer cell lines that yield high titers of LGSN vector in supernatants were developed by liposome-mediated transfection in combination with cross infection. Both GP + envAm12 murine fibroblasts and K562 leukemic cells transduced with EGFP virus demonstrated a stable green fluorescence signal readily detectable by flow cytometry or fluorescence microscopy in up to 97% and 86% of examined cells, respectively. The integration of LGSN provirus in transduced cells was confirmed by PCR analysis. These results indicate EGFP is a suitable reporter molecule for gene transfer and expression in hematopoietic cells. Therefore, the bright and long-term expression of EGFP in living cells will advance the study of gene therapy in vitro and in vivo, particularly for human applications.