Objective In the candidate region 12q13 of simple congenital heart defect(CHD),two single nucleotide polymorphisms(SNPs)in the coding-region of GLI1 gene,rs11553626 and rs2228226,were chosen to investigate their distribution in 180 simple CHD patients and 200 normal controls which were collected in the first affliated hospital,China Medical University and The General Hospital of Shenyang Military Area,in order to determine the relationship between GLI1 gene and simple CHD.Methods Genotypes of these two SNPs were analyzed in 180 simple CHD patients and 200 normal controls by Denatured High Performance Liquid Chromatography(DHPLC)and sequencing from Jan.2000 to Jun.2005.?~2 test was applied to analyze the genotype frequency and allele frequency between CHD groups and control groups.Results No polymorphisms were found at rs11553626 while G/C polymorphisms were found at rs2228226.Remarkable significance was observed at rs2228226 in the allele frequency distribution(?~2=8.956,P

ObjectiveTo investigate the association between R405Q polymorphism of GLI1 gene and tetralogy of fallot(TOF).Methods In the case-control study,the R405Q polymorphism of GLI1 gene in 112 children with TOF and 200 healthy controls were detected with polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).The distribution of genotype and allele frequency at R405Q polymorphism site were analyzed and to investigate its relationship with the risk of TOF.ResultsThe distribution of genotype frequency at R405Q polymorphism site was not different between TOF group and the healthy control group(x2 =5.317 ,P = 0.07) .However, the distribution of allele frequency at R405Q polymorphism site was significantly different between TOF group and the healthy control group (x2 = 6.790, P = 0.009) , and the relative risk for TOF in A allele carriers was higher than that in G allele carriers (OR = 1.561,95％ CI 1.116 ～ 2.185) Conclusion The R405Q polymorphism of GLI1 gene is associated with TOF and people with A allele have higher risk with TOF.GLI1 gene might be the genetic susceptibility gene of TOF.

OBJECTIVETo identify and characterize laryngeal cancer related novel genes located on chromosome 6q25.

METHODSElectric hybridization was performed in human genome database using EST (expression sequence tag) as probe. Novel genes were deduced by software from positive DNA clones and their cDNAs were amplified by RT-PCR using primers designed according to the sequence of the putative genes.

RESULTSA novel gene was cloned successfully. The full length of this gene was about 21 kb. It contained two exons and produced a 1006 bp transcript coding a protein with 235 amino acid residues. It's 5'flanking sequence contained two binding sites of oncoprotein c-Myc, thus it was named MTLC (c-Myc target from laryngeal cancer cells). Homologous assay showed that MTLC exhibited little overall homology to known human proteins but it exhibited good overall homology to mouse MT-MC1 protein with an identity of 78%. The primary structure of MTLC protein contained a nuclear location signal motif, but it did not have other conserved domains. The results of subcellular location experiment showed that MTLC expressed in nuclei of human hepatocellular carcinoma cell line Bel7402 cells, while a wide distribution of MTLC in various tissues was demonstrated by Northern blotting.

CONCLUSIONMTLC may play an important role as a target gene of c-Myc and as a transcription factor in keeping the normal physiological process of cells.

Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; Chromosomes, Human, Pair 6 ; genetics ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Gene Expression ; Green Fluorescent Proteins ; Humans ; Laryngeal Neoplasms ; genetics ; Luminescent Proteins ; genetics ; metabolism ; Microscopy, Fluorescence ; Molecular Sequence Data ; Nuclear Proteins ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Tumor Cells, Cultured

Objective: To analyze the current situation and needs of primary and secondary school health clinic services, and to provide a reference for strengthening school health service in China. Methods: Investigation provinces were selected according to China s three major economic zones, 1 or 2 survey centers were selected from each province, 16 provinces and 26 survey centers were selected. In each survey centers, six schools from primary and secondary schools, junior high schools and high schools in urban and rural areas were randomly selected as research sites. Each research site randomly selected 10 students in each grade(grade three, four, five from primary schools) to fill in the questionnaire anonymously. A total of 4 661 valid questionnaires were collected. Results: Among the services provided by the school clinics to students, "trauma treatment" "consulting health problems" "physical examination" accounted for 49.4%, 40.5% and 39.0% respectively. Within the past one year, 45.1% students went to the school clinic for consultation or consultation 1-3 times, 6.4% for 4-5 times, and 3.7% for 6 times or more. When students were sick in school, 65.8%(primary school), 64.0%(junior high school), and 54.4%(high school) of them selected to go to the school clinic. When suddenly suffered trauma in school, 78.2%(primary school), 73.2%(junior high school) and 69.6%(high school) of the students went to the school clinic. Conclusion Primary and middle school students have a great demand for school health services, and it is necessary to continue to strengthen the construction of primary and secondary school health clinics.

Objective: To investigate the effect of silencing troponin I3 (Tnni3) gene expression on biological property of rat embryonic H9C2 cardiomyocytes. Methods: The rat embryonic H9C2 cardiomyocytes were cultured and divided into 2 groups: control group transfected with negative control small interfering RNA (NC-siRNA group) and experimental group transfected with Tnni3 small interfering RNA (Tnni3-siRNA group). At 48 h, 72 h after transfection, the cells were collected, and real time quantitative polymerase chain reaction (qPCR)was used to detect the mRNA expressions of Tnni3 and Caspase-3, and Western blot was used to detect the protein expressions of Tnni3, Cyclin A1 and Cyclin B1.Annexin V-fluorescein isothiocyanate(FITC) apoptosis detection kit was used to analyze cell apoptosis.Cell proliferation was measured by Cell Counting Kit-8 (CCK-8) solution and cell cycle was detected by flow cytometry. Results: At 48 h post-transfection with Tnni3-siRNA, H9C2 cells exhibited a significant decrease in Tnni3 mRNA (0.27±0.05 vs. 1.00±0.00) and protein (0.18±0.03 vs. 1.00±0.00) compared with those transfected with NC-siRNA, and the differences were statistically significant (t=25.26, 47.40, all P<0.01). Apoptotic cells were observed in the NC-siRNA group and the Tnni3-siRNA group.At 72 h post-transfection, the percentage of apoptotic cells significantly increased in H9C2 cells transfected with Tnni3-siRNA [(11.30±1.85)% vs. (0.33±0.15)%] compared with those transfected with NC-siRNA, an increased expression of Caspase-3 mRNA was also observed in Tnni3-siRNA-transfected H9C2 cells (1.39±0.13 vs. 1.00±0.00), and the differences were statistically significant (t=10.24, 5.19, all P<0.01). Compared with NC-siRNA-transfected H9C2 cells, a time-dependent reduction in cell proliferation was observed in Tnni3-siRNA-transfected H9C2 cells (48 h: 0.32±0.06 vs. 0.46±0.03; 72 h: 0.31±0.01 vs. 0.63±0.04; 96 h: 0.36±0.01 vs 0.75±0.04), and the differences were statistically significant (t=3.62, 13.45, 16.39, all P<0.01). At 72 h post-transfection with Tnni3-siRNA, the percentage of G1 phase, S phase and G2 phase cells was (71.25±3.82)%, (18.28±2.78)% and (9.94±1.09)%, respectively.There was a significant increase in the proportion of G2 phase cells [(9.94±1.09)% vs. (4.54±0.99)%] in H9C2 cells transfected with Tnni3-siRNA compared with those transfected with NC-siRNA, an increased expression of Cyclin A1 protein (1.89±0.09 vs.1.00±0.00) and a decreased expression of Cyclin B1 protein (0.47±0.06 vs.1.00±0.00) were observed in Tnni3-siRNA-transfected H9C2 cells, respectively, and the differences were statistically significant (t=6.35, 17.12, 15.32, all P<0.01). Conclusions Silencing Tnni3 gene expression in rat embryonic H9C2 cardiomyocytes can induce cell apoptosis, suppress cell proliferation, and led to G2 cell cycle arrest.

Plant polyphenols are phenylpropanoid derivatives including phenolic acids, stilbenes, curcumins and flavonoids. These compounds display a variety of biological and pharmacological activities such as antioxidation, vasorelaxation, anti-coagulation, anti-inflammation, anti-tumor and anti-virus, conferring a huge application potential in the sectors of drugs, foods, cosmetics, and chemicals. Microorganisms have become important hosts for heterologous synthesis of natural products due to the advantages of fast growth, easiness of culture and industrial operation. In recent years, the development of synthetic biology has boosted the microbial synthesis of plant natural products, achieving substantial progress. In this review, we summarize the synthesis of plant polyphenols in engineered Escherichia coli, Saccharomyces cerevisiae and other microorganisms equipped with the designed biosynthetic pathways of polyphenols. We also discuss the optimization strategies such as precursor engineering, dynamic regulation, and co-cultivation to improve the production of polyphenols and propose future prospects for polyphenol pathway engineering.
Biosynthetic Pathways ; Metabolic Engineering ; Plants ; Polyphenols ; Saccharomyces cerevisiae/genetics*