Objective To screen the 5’ regulatory region of the aldose reductase (AR) gene for genetic variabilities causing changes in protein expression and affecting the promoter function. Methods The screenings were carri...Objective To screen the 5’ regulatory region of the aldose reductase (AR) gene for genetic variabilities causing changes in protein expression and affecting the promoter function. Methods The screenings were carried out by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). All SSCP variants were submitted for DNA sequencing and inserted into the plasmid chloromycetin acetyl transferase (CAT) enhancer vector. The constructs were used to transfect Hela cells,and CAT assays were performed to assess promoter activity. Gel mobility shift and footprinting assays were also performed to determine the interaction between the DNA and nuclear proteins. Results Two polymorphisms, C(-106)T and C(-12)G, were identified in the regulatory region in 123 Chinese control subjects and 145 patients with type 2 diabetes mellitus. The frequencies of genotypes WT/WT, WT/C(-12)G and WT/C(-106)T were not significantly different between the subjects and patients. In the patients with and without retinopathy, frequencies of WT/C(-106)T were 31.5% and 17.5% (P【0.05) respectively, and the frequencies of WT/C(-12)G were 10.5% and 2.5% (P】0.05) respectively. The total frequency of WT/C(-12)G and WT/C(-106)T in patients with retinopathy was 41.8%, significantly higher than that (20.0%) in patients without retinopathy (P【0.025). The relative transcription activities of the wild-type, the C(-12)G and the C(-106)T were 15.7%, 31.0% and 32.2%, respectively. The results of DNA-protein interaction assays showed that these variations did not change the binding site of DNA with trans-acting factors. Conclusion The polymorphisms C(-12)G and C(-106)T strongly associated with diabetic retinopathy in the Chinese population have been identified in the regulatory region of the aldose reductase gene.展开更多
AIM： To present the rationale, design, methodology, and the baseline data of the Beijing Desheng Diabetic Eye Study（BDDES）, and to determine the prevalence of diabetic retinopathy（DR） and possible risk factors in...AIM： To present the rationale, design, methodology, and the baseline data of the Beijing Desheng Diabetic Eye Study（BDDES）, and to determine the prevalence of diabetic retinopathy（DR） and possible risk factors in patients with type 2 diabetes mellitus（T2DM） in an urban community of Beijing, China.METHODS： Community-based prospective cohort study of persons diagnosed with T2DM aged 30 y or older. The main variables of interest are the presence and progression of DR as determined by the standardized ETDRS grading of seven fields fundus photographs. The presence and severity of DR were analyzed for possible correlations to non-genetic and genetic dispositions.RESULTS： A total of 1438 participants with data available for analysis, the prevalence of any DR was 35.4%. The prevalence of mild non-proliferative diabetic retinopathy（NPDR）, moderate NPDR, severe NPDR, and proliferative diabetic retinopathy was 27.7%, 2.6%, 0.5% and 4.5%, respectively. By multiple logistic regression analysis, risk factors for the presence of any DR included male（P=0.031）, lower income level（P=0.011）, lower education background（P=0.022）, longer duration of diabetes（P=0.001）, younger age at diabetic onset（P=0.001）, higher systolic blood pressure（P=0.007）, higher glycosylated hemoglobin A1 c levels（P=0.001）, high albuminuria（P=0.03）, and use of insulin（P〈0.001）. For vision-threatening DR, four factors were significant： younger age at diabetic onset（P〈0.001）,higher systolic blood pressure（P=0.042）, high albuminuria（P〈0.001）, and use of insulin（P〈0.001）. CONCLUSION： The BDDES is the first large-scale ongoing cohort study of a Chinese urban population of persons with type 2 diabetes. Using standardized grading system comparable to large cohort studies from western populations, our baseline data shows that the prevalence of DR and major risk factors in this Chinese ethnic population are comparable to that found in the western population studies.展开更多
基金grantsfromtheNationalNaturalScienceFoundationofChina (No 396 70 35 2 )
文摘Objective To screen the 5’ regulatory region of the aldose reductase (AR) gene for genetic variabilities causing changes in protein expression and affecting the promoter function. Methods The screenings were carried out by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). All SSCP variants were submitted for DNA sequencing and inserted into the plasmid chloromycetin acetyl transferase (CAT) enhancer vector. The constructs were used to transfect Hela cells,and CAT assays were performed to assess promoter activity. Gel mobility shift and footprinting assays were also performed to determine the interaction between the DNA and nuclear proteins. Results Two polymorphisms, C(-106)T and C(-12)G, were identified in the regulatory region in 123 Chinese control subjects and 145 patients with type 2 diabetes mellitus. The frequencies of genotypes WT/WT, WT/C(-12)G and WT/C(-106)T were not significantly different between the subjects and patients. In the patients with and without retinopathy, frequencies of WT/C(-106)T were 31.5% and 17.5% (P【0.05) respectively, and the frequencies of WT/C(-12)G were 10.5% and 2.5% (P】0.05) respectively. The total frequency of WT/C(-12)G and WT/C(-106)T in patients with retinopathy was 41.8%, significantly higher than that (20.0%) in patients without retinopathy (P【0.025). The relative transcription activities of the wild-type, the C(-12)G and the C(-106)T were 15.7%, 31.0% and 32.2%, respectively. The results of DNA-protein interaction assays showed that these variations did not change the binding site of DNA with trans-acting factors. Conclusion The polymorphisms C(-12)G and C(-106)T strongly associated with diabetic retinopathy in the Chinese population have been identified in the regulatory region of the aldose reductase gene.
基金Supported by the Beijing Natural Science Foundation(No.7131007)the Norwegian Research Council(No.180419/D15/1k)
文摘AIM： To present the rationale, design, methodology, and the baseline data of the Beijing Desheng Diabetic Eye Study（BDDES）, and to determine the prevalence of diabetic retinopathy（DR） and possible risk factors in patients with type 2 diabetes mellitus（T2DM） in an urban community of Beijing, China.METHODS： Community-based prospective cohort study of persons diagnosed with T2DM aged 30 y or older. The main variables of interest are the presence and progression of DR as determined by the standardized ETDRS grading of seven fields fundus photographs. The presence and severity of DR were analyzed for possible correlations to non-genetic and genetic dispositions.RESULTS： A total of 1438 participants with data available for analysis, the prevalence of any DR was 35.4%. The prevalence of mild non-proliferative diabetic retinopathy（NPDR）, moderate NPDR, severe NPDR, and proliferative diabetic retinopathy was 27.7%, 2.6%, 0.5% and 4.5%, respectively. By multiple logistic regression analysis, risk factors for the presence of any DR included male（P=0.031）, lower income level（P=0.011）, lower education background（P=0.022）, longer duration of diabetes（P=0.001）, younger age at diabetic onset（P=0.001）, higher systolic blood pressure（P=0.007）, higher glycosylated hemoglobin A1 c levels（P=0.001）, high albuminuria（P=0.03）, and use of insulin（P〈0.001）. For vision-threatening DR, four factors were significant： younger age at diabetic onset（P〈0.001）,higher systolic blood pressure（P=0.042）, high albuminuria（P〈0.001）, and use of insulin（P〈0.001）. CONCLUSION： The BDDES is the first large-scale ongoing cohort study of a Chinese urban population of persons with type 2 diabetes. Using standardized grading system comparable to large cohort studies from western populations, our baseline data shows that the prevalence of DR and major risk factors in this Chinese ethnic population are comparable to that found in the western population studies.
