Zinc transporter 8 (ZnT8) is an important candidate antigen for type Ⅰ diabetes. The autoantibody detection kit based on ZnT8 can be used to help diagnose type Ⅰ diabetes, and the related products have been launched in Europe and the United States. Since the recombinant production system of active ZnT8 has not been established in China, this key raw material is heavily dependent on imports. We used Saccharomyces cerevisiae to carry out the recombinant expression of ZnT8. First, multiple antigenic forms of ZnT8 were designed as C-terminal haploid (C), C-terminal diploid (C-C), and N-terminal and C-terminal concatemers (N-C). The proteins were expressed, purified and tested for antigenicity by bridging-type ELISA. The serum of 13 patients with type Ⅰ diabetes and the serum of 16 healthy volunteers were detected. C, N-C, and C-C proteins had similar detection rates, which were 53.8% (7/13), 61.5% (8/13) and 53.8% (7/13). The specificity of the three groups was 100% (16/16). The detection value on positive samples P3, P4, and P8 increased by more than 90%, indicating better serum antibody recognition ability. Finally, N-C protein was selected for further serum sample testing, and the test results were characterized by receiver operating characteristic (ROC) curve for sensitivity and specificity. Compared with imported gold standard antigen, the sensitivity was 76.9% (10/13) and the specificity was 87.5% (14/16). There was no significant difference in the sensitivity of the method, but the specificity needed to be improved. In conclusion, the ZnT8 N-terminal and C-terminal concatemer protein developed based on S. cerevisiae expression system is expected to be a key alternative raw material in the development of in vitro diagnostic reagents for type Ⅰ diabetes.
Antigens ; Autoantibodies ; Diabetes Mellitus, Type 1/diagnosis* ; Enzyme-Linked Immunosorbent Assay ; Humans ; Saccharomyces cerevisiae/genetics* ; Zinc Transporter 8/genetics*

OBJECTIVETo explore the relationship between the polymorphism of solute carrier family 30, member 8 (SLC30A8) gene and type 2 diabetes mellitus (T2DM) in Uyhgur in Xinjiang and further analyze the interaction between SLC30A8 gene polymorphism loci and smoking.

METHODSA case control study, including 1 000 patients with T2DM and 1 010 non-diabetic controls, was conducted in Xinjiang. All the subjects were Uygur and the age difference between the two groups was within 3 years. Physical examination and blood biochemical detection were performed to obtain personal clinical parameters. Genomic DNA was extracted from peripheral blood leukocytes. The single nucleotide polymorphism (SNP) of SLC30A8 of all the subjects was tested by using MALDI-TOF. Statistical analyses were performed with SPSS 16.0. Bootstrap method was used to calculate 95% confidence intervals of RERI, AP and S.

RESULTSAfter adjusting BMI, SBP, TC, HDL-C and LDL-C, rs13266634 of SLC30A8 gene genotype frequency and allele frequency distribution had statistical differences (P<0.05). Rs13266634 of risk allele were C, OR was 1.194 (95% CI: 1.044-1.366). In addition, the data from genotype distribution analysis under different models showed that significant association between rs13266634 and T2DM in dominant model, OR was 1.640 (95% CI 1.072-2.510). The product of rs13266634 with the active smoking or passive smoking had no statistical significance (P>0.05) , indicating there were no multiplication interaction among them. Additive interactions index of RERI, AP and S and its 95% confidence interval of rs13266634 and active smoking, rs13266634 and passive smoking were 0.301 (-1.314-0.712), 0.204 (-0.854-0.446), 0.612 (0.186-2.013) and 0.125 (-0.805-1.055), 0.052 (-0.353-0.456), 1.096 (0.500-2.403) respectively, indicating there were no significant additive interaction among them.

CONCLUSIONRs13666334 of SLC30A8 gene is associated with the susceptibility of T2DM in Uygur, and its protective genotype might be TT. Passive smoking might increase the risk of T2DM in Uygur.

Alleles ; Case-Control Studies ; Cation Transport Proteins ; genetics ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; epidemiology ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Single Nucleotide ; Risk Factors ; Smoking ; epidemiology ; Zinc Transporter 8

OBJECTIVETo investigate a potential relationship between Solute carrier family 30 (zinc transporter) member 8 (SLC30A8) rs13266634 variant and efficacy of rosiglitazone or repaglinide in treating newly diagnosed Chinese type 2 diabetes patients.

METHODSA total of 209 diabetic patients without any antihyperglycemic history were recruited and treated with repaglinide or rosiglitazone randomly for 48 weeks (104 and 105 patients, respectively). Anthropometric measurements and clinical laboratory tests were carried out before and after the treatment. An non-synonymous variant rs13266634 was genotyped by matrix-assisted laser desorption ionization-time of flight mass spectroscopy.

RESULTSNinety-one patients in repaglinide group and ninety-three patients in rosiglitazone group completed the study. Δ value of homeostasis model assessment of beta cell function (HOMA-B) and Δ value of fasting proinsulin levels were statistically significant between three genotype groups (P=0.0149 and 0.0246, respectively) after rosiglitazone treatment. However, no genotype association was observed in the repaglinide or rosiglitazone group with other parameters.

CONCLUSIONThe SLC30A8 variant was associated with the efficacy of insulin sensitizer monotherapy on insulin secretion in patients with newly diagnosed type 2 diabetes mellitus in Shanghai, China.

Carbamates ; therapeutic use ; Cation Transport Proteins ; genetics ; China ; Diabetes Mellitus, Type 2 ; drug therapy ; genetics ; Female ; Gene Frequency ; Humans ; Hypoglycemic Agents ; therapeutic use ; Male ; Middle Aged ; Piperidines ; therapeutic use ; Polymorphism, Single Nucleotide ; Thiazolidinediones ; therapeutic use ; Zinc Transporter 8

OBJECTIVETo study the associations of single nucleotide polymorphisms (SNPs) of TCF7L2, CDKAL1, SLC30A8, HHEX with diabetic retinopathy (DR) and nephropathy (DN) in type 2 diabetes mellitus.

METHODSA total of 479 subjects with DR,248 with DN and 650 without DR or DN were recruited to assess the associations between SNPs of TCF7L2 (rs7903146, rs6585205, rs11196218), CDKAL1 (rs10946398,rs4712527), SLC30A8 (rs13266634, rs3802177, rs11558471) and HHEX (rs1111875, rs7923837) and the development of DR and DN.

RESULTSThere were significant differences in genotypic and allele frequencies of rs11558471 (SLC30A8) between DR and control groups (P< 0.05), the odds ratio (OR) values of A and AA were 1.27 and 1.68. The distributions of genotype and allele frequency for rs11196218 (TCF7L2) were significantly different between DN and control group (P=0.0051,OR=1.37). However, the P value after Bonferroni correction showed no significant difference. No significant differences were found in the distributions of rs13266634 and rs3802177 (SLC30A8), rs10946398 (CDKAL1), rs6585205, rs7903146 and rs11196218 (TCF7L2) and rs7923837 (HHEX) between DR and control groups, and nor significant differences were found in distributions of rs6585205 (TCF7L2), rs4712527 (CDKAL1), rs13266634, rs3802177 and rs11558471 (SLC30A8), and 7923837 (HHEX) between DN and control groups, though for all comparison the OR values were greater than 1.

CONCLUSIONPolymorphisms of SLC30A8 and TCF7L2 genes may be associated with the development of DR and DN, respectively. Association between the polymorphisms of CKDAL1, TCF7L2 and HHEX genes and DR, and between the polymorphisms of SLC30A8, HHEX and CDKAL1 genes and DN, cannot be excluded.

Cation Transport Proteins ; genetics ; Cyclin-Dependent Kinase 5 ; genetics ; Diabetes Mellitus, Type 2 ; complications ; genetics ; Diabetic Angiopathies ; genetics ; Female ; Homeodomain Proteins ; genetics ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Transcription Factor 7-Like 2 Protein ; genetics ; Transcription Factors ; genetics ; Zinc Transporter 8 ; tRNA Methyltransferases