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Objective To explore the surface morphological and biomechanical properties differences of peripheral blood mononuclear cells (PBMCs) between groups of patients with mitochondrial diabetes caused by mt.3243A ＞ G mutation and healthy controls.Methods 2 milliliters blood were obtained from each subject of the mitochondrial diabetes group (n =5) and the control group (n =5).The PBMCs were separated from the blood using the standard Ficoll-Hypaque density-gradient centrifugation method and detected by atomic force microscope (AFM).Results The morphological analysis revealed that compared with control group,the PBMCs of diabetic patients tended to have a lower cell height (0.73 ± 0.24μm vs 2.49 ± 1.17μm,P =0.011) and a much rougher cell membrane (Ra:161.8 ± 33.2nm vs 66.4 ± 16.3 nm,P =0.000;Rq:202.2 ± 40.9nm vs 85.4 ± 17.1 nm,P =0.000).The adhesion force distribution was nearly three times higher in PBMCs of diabetic patients than that of the control group (779.6 ± 190.0pN vs 161.1 ± 83.1 pN,P =0.000).The Young's modulus of PBMCs was significantly increased in diabetic patients (421.4 ± 140.0kPa vs 138.3 ± 77.2kPa,P ＜ 0.01),indicating that diabetic PBMCs were stiffer than control cells.Conclusion Our study demonstrated the surface morphological and biomechanical properties changes in mitochondrial diabetes caused by mt.3243A ＞ G mutation at PBMCs level,which was beneficial to the better understanding of the pathophysiological mechanisms of mitochondrial diabetes associated with mt.3243A ＞ G mutation.
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Objective Assessing the detection performance of testing mycoplasma pneumonia(MP) type-specific antibodies by Chemiluminescence immunoassay(CLIA), in order to evaluate the feasibility of screening MP infection by CLIA.Methods Total of 280 cases of respiratory disease patients,20 examples infected mycoplasma pneumonia and 20 cases health volunteers as the control group were enrolled in this study from August 2016 to October 2017 in the Nanfang Hospital,Southern Medical University,testing MP antibodies by CLIA,Enzyme linked immunosorbent assay(ELISA)and Passive agglutination method(PA) respectively.According to the performance evaluation scheme, we evaluate the performance indexs of detecting MP antibodies by CLIA, including lower limit of detection, intra-batch precision, inter-batch precision,linearity range,clinical coincidence rate and consistency compared with ELISA and PA,and the results were analyzed by EXCEL and SPSS version 22.0.Results MP-IgG CLIA reagent:Limit of blank, Limit of detection and Limit of quantitation were 1.9 AU/ml,4.5 AU/ml and 5.1 AU/ml respectively;Coefficient Variation(CV)of intra-batch precision in high and low concentration levels were 2.98% and 2.45%respectively; CV of inter-batch precision in high and low concentration levels were 6.44% and 6.83% respectively;both the Linear range and Clinical report range are from 2.0 AU/ml to 253.0 AU/ml;the linear regression equation R 2≥0.990 0,0.85≤b≤1.15.MP-IgM CLIA reagent: CV of intra-batch precision in high and low concentration levels were 2.55% and 2.86% respectively; CV of inter-batch precision in high and low concentration levels were 4.82% and 5.46% respectively.The total clinical coincidence rate of MP-IgG and MP-IgM detected by CLIA were 90.0%and 97.5%respectively.The kappa values of MP-IgG and MP-IgM detected by CLIA and ELISA were 0.763(P=0.000)and 0.804(P=0.023)respectively, with Consistent percentage of 88.9% and 91.4% respectively.The kappa value of CLIA and PA was 0.541(P=0.063)with a consistent percentage of 79.6%.Conclusions The results of study show that detecting MP type-specific antibodies by CLIA meet the prescribed performance indexes. Detecting MP type-specific antibodies by CLIA,which is precise, speedy and automated, could be applied to clinical and replace ELISA and PA, becoming the prior choice in clinical for MP infection screening.
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<p><b>OBJECTIVE</b>To establish a rapid, accurate, noninvasive and low cost method for screening MT3243A>G mutation in mitochondrial diabetes.</p><p><b>METHODS</b>Blood, saliva, and urine sediment samples were collected from 6 patients with confirmed mitochondrial diabetes and 50 healthy controls from Shanghai Children's Hospital and Shanghai Sixth People's Hospital. The heterozygosity levels of MT3243A>G mutation in above samples were detected with pyrosequencing, and the data were compared. MT3243A>G mutations were rapidly screened with high resolution melting curve analysis (HRM) in the urine sediment samples of 1070 diabetic patients from 4 communities in Shanghai. Furthermore, pyrosequencing was used to validate the suspected positive samples, and the heterozygosity levels were also quantified.</p><p><b>RESULTS</b>Comparative experiments found that heterozygosity of MT3243A>G mutation was 2 to 7 times higher in urine sediment than in saliva and blood samples from the 6 patients with confirmed mitochondrial diabetes. However, the heterozygosity was slightly higher in saliva than blood samples. MT3243A>G mutation was not detected in the 50 healthy controls. Two samples with suspected MT3243A>G mutation were identified in the 1070 urine sediment samples of diabetes patients with HRM screening, which were validated by pyrosequencing. The heterozygosity of MT3243A>G mutation were 33.32% and 14.67% in the urine sediment samples, respectively.</p><p><b>CONCLUSION</b>Urine sediment samples can be used for rapid screening of MT3243A>G mutation for its ease to collect, noninvasiveness and higher level of heterozygosity. HRM is suitable for rapid screening for mitochondrian mutations for its low cost, while such mutations could be detected with sensitivity and accuracy by pyrosequencing.</p>
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Humans , DNA, Mitochondrial , Genetics , Diabetes Mellitus , Genetics , Heterozygote , Mutation , Sequence Analysis, DNA , Methods , Transition TemperatureABSTRACT
Metabolic diseases information consulting provides its clients with data and insight to help translation information medicine product development plans:for clinical,diagnosis,biologics,medical devices/combination products.Understanding the mechanisms by which specific protein functions contribute to metabolic disease pathogenesis is a great challenge.The barcode is the common nominator and identifier of a sample.This code can be utilized in both 2D form,capturing important identifiers for each sample type and origin.Visual database informational system,to impact on the quality of the analysis data generated.Our ultimate motivation lies in helping you to advance the translation medicine success of our hospital,and to optimize the benefit they provide to patients in need.
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Objective To compare the differences of metabolic syndrome (MS) prevalence by using four working definitions and their relationship with obesity-related indicators in first-degree relatives of type 2 diabetes mellitus pedigrees. Methods Totally, 2 372 first-degree relatives from 715 type 2 diabetic pedigrees were selected in this study. Complete laboratory data, including blood pressure, lipid profile and plasma glucose, were collected. The prevalence rates of MS and obesity of four definitions, as defined by National Cholesterol Education Program Adult Treatment Panel Ⅲ (ATPⅢ) in 2005, International Diabetes Federation (IDF) in 2005,Chinese Diabetes Society (CDS) in 2004 aml Joint Committee for Developing Chinese Guidelines on Prevention and Treatment of Dyslipidemia in Adults (JCDCG) in 2007,were analyzed. Results (1)The prevalence rates of MS were 45.40% ,38.74% ,25.08% and 39.29% aecording to four definitions respectively. The prevalence rates of MS were higher in females than in males by using ATPⅢ and IDF definitions (both P<0. 01). (2)The common comhinations of metabolic abnormality was dyslipidemia, hypertension, obesity and hyperglycemia by using four definitions,except in females by using CDS definition. (3)The prevalence rates of obesity were 58.18% ,58.18% ,33.90% and 42.96% acconling to the four definitions respectively. The prevalence rates of MS in obese subjects were 66.59% ,66.59% ,54.85% and 68.99% according to four definitions respectively. (4) Applying the cutoff point for abdominal obesity according to ATPⅢ, IDF and JCDCG definitions, the prevalence rates of abdominal obesity in subjects with body mass index (BMI) <25 kg/m2 were respectively 28.58% and 16.78%, being higher in females than in males(38.90% vs 15.02% ,21.01% vs 11.22% ,both P<0. 01). Conclusion (1)There is significant familial aggregation of MS and obesity,and the first-degree relatives of type 2 diabetic patients are high risk populations. (2) Waist circumference rather than BMI taken as a discriminating component of obesity in MS seems to be clinically more helpful to the early identification and prevention of MS.
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Objective To investigate the effect of solute cartier organic anion transporter family, member 1B1 (SLCOIBI) gene variants on the response to therapy with repaglinide in type 2 diabetes. Methods 100 newly-diagnosed type 2 diabetic patients were treated with repaglinide during a course of 48 weeks. Anthropometrie parameters and indices related to glucose metabolism were measured periodically. Genotypes of SLCO1B1 D130N and V174A were detected by PCR-restriction fragment length polymorphism (RFLP) and sequencing respectively. Results Eighty-nine patients accomplished the 48-week follow-up visits. D130N variant in SLCO1B1 gene was associated with repaglinide treatment, DD genotype had better HbA1C lowering effect than N allele carrier [△HbA1C: (-2.29±0.23) % vs (-1.49±0.21)%, P<0.05]. No association was detected between D130N and the other effects of repaglinide on glucose metabolism related phenotypes. Conclusion D130N variant in SLCO1B1 gene is associated with the response to repaglinide treatment in patients with type 2 diabetes. DD homozygotes had a better effect than N allele carriers.
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To explore the association between alkaline phosphatase (ALP) and degenerative cardiac valvular calcification, the activity of ALP and the grades 0～Ⅳ degenerative calcified valves were determined by biochemical and immunocytochemical methods. ALP positive substance was present in the matrix around the foci of calcification and in the cytoplasm of fibroblasts. The bioactivity of ALP was much higher in grades Ⅰ and Ⅱ calcified valves than that in grades 0 and Ⅲ～Ⅳ calcified valves. The results indicate that the level of ALP bioactivity is predominantly dependent on the process of extruding matrix vesicles from senescent cells in the valves.
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Objective To investigate the association of activating transcription factor 6(ATF6)gene Ala145Pro(GCG→CCG)variant with glucose and lipid metabolism in Chinese.Methods The genotypes were determined by PCR-RFLP in 689 Chinese in Shanghai.Among them,361 subjects showed normal glucose regulation,250 cases were newly-diagnosed diabetic patients without taking any drug and 78 cases were probands of early-onset type 2 diabetes pedigrees.The following phenotypes were measured:body height and weight to calculate BMI;waist,hip and femoral circumference to calculate waist-to-hip ratio and waist-to-femoral ratio;blood pressure;plasma glucose levels obtained at 0 and 120 minute during 75 g oral glucose tolerance test;fasting serum lipid profile including total cholesterol,triglyceride,high density lipoprotein-cholesterol and low density lipoprotein-cholesterol;body fat percentage and distribution.Results(1)The frequency of C allele is significantly lower in probands from early-onset type 2 diabetes patients compared with subjects with normal glucose regulation(P=0.035).(2)In subjects with normal glucose regulation,the CC+CG genotype had a significantly lower level of high density lipoprotein cholesterol as compared with GG genotype(P=0.014).(3)In type 2 diabetic patients,the CC+CG genotype had a significantly higher level of low density lipoprotein-cholesterol as compared with GG genotype(P=0.041).Conclusion These findings suggest that variant of ATF6 plays a role in glucose and lipid metabolism in Chinese.