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Objective:To explore the clinical characteristics and follow-up outcomes of a pedigree of maturity onset diabetes of the young (MODY) induced by a novel mutation of glucokinase (GCK).Methods:The clinical features and laboratory data of a pedigree diagnosed with GCK-MODY in Peking Union Medical College Hospital was analyzed. Genomic DNA was extracted, and Sanger sequencing was performed to detect the gene mutation of the family members. The proband and her father were followed up for 3 years. Wanfang and PubMed were used to search literatures on follow-up studies for treatment of GCK-MOYD.Results:Both the proband and her father were found to have a novel mutation on the GCK gene located in exo10 c.1348G.T (p. Ala450Thr). The proband was treated with diet and exercise control only. At the end of the follow-up, her fasting plasma glucose (FPG, 6.8 mmol/L), 2 h postprandial plasma glucose (2hPG, 7.4 mmol/L), and glycated hemoglobin (HbA1c, 6.3%) were all within the control targets. Additionally, the levels homeostasis model assessment of insulin resistance (HOMA-IR) tended to improved comparing to that at baseline (4.09 to 2.32), and glucose disposition index (DI) was improved compared with baseline (16.22 to 20.05). As to the proband′s father, the treatment with insulin plus acarbose was converted to sulfonylureas monotherapy. His FPG and 2hPG mostly were within the target range, and the levels of HbA1c were significantly reduced by 0.5%-0.7% when compared to that at baseline. The HOMA-IR or islet beta cell function was comparable to those at baseline.Conclusions:Screening patients whose clinical performance meets GCK-MODY and their family members with proper genetic testing is of great importance to reduce misdiagnosis of GCK-MODY, so as to obtain a better glucose control without unnecessary over-treatment and protect islet beta cell function.
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Objective Cloning and expression of Par6A.Methods Par6A cDNA was amplified from rat L6 skeletal muscle cells by RT-PCR and the cloning and expression vectors of Par6A were constructed.The expression vector was transfected into 293 cells.Furthermore,the function of Par6A was confirmed by Co-immunoprecipitation.Results Par6A cDNA with approximately 1 kb in length was successfully amplified,and the expression vector of pDsRed-Express-N1-Par6A was constructed.The red fluorescene was seen under fluorescent microscope after 293ET cells were transfected for 24 h using the pDsRed-Express-N1-Par6A vector.The expressed Par6A protein can interacte with PKC?.Conclusion We successfully cloned the Par6A cDNA from rat L6 skeletal muscle cells,which provided a reliable method to study the function of Par6A.
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Objective To construct human SREBP-1c-promoter reporter gene vector and to detect its function.Methods Human blood genome DNA was extracted and pGL3-Basic-SREBP-1c-promoter reporter gene vector was constructed.Furthermore,the function of SREBP-1c-promoter was confirmed by dual-luciferase reporter assay.ResultspGL3-Basic-SREBP-1c-promoter reporter gene vector was successfully constructed and the promoter activity was obviously repressed by co-transfection FoxO1.Overexpression FoxO1 inhibited the SREBP-1c protein expression.Conclusion FoxO1 repressed the SREBP-1c protein expression through inhibition the SREBP-1c transcription.
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<p><b>OBJECTIVE</b>To illuminate the regulating effect of all-trans retinoic acid (ATRA) on gap junctional intercellular communication (GJIC) and connexin 43 (Cx43) gene expression in glioma cells, which is tissue- and organ-specific.</p><p><b>METHOD</b>Rat C6 glioma cells were exposed to ATRA at a concentration of 1, 10, 100 micromol/L and the GJIC function of the cells was examined with scrape-loading dye transfer assay 24 hours, 48 hours and 72 hours after ATRA treatment. The effect of ATRA on Cx43 gene expression was measured with semiquantitative reverse transcription polymerase chain reaction (RT-PCR) 24 hours after ATRA exposure.</p><p><b>RESULTS</b>The GJIC function of C6 glioma cells was significantly increased by ATRA at each concentration applied. The dye passed 4 to 5 rows of cells from the scraping edge in ATRA treated cells, but only 1 or 2 rows in the control. The augment effect was observed 24 hours after each concentration ATRA treatment, and lasted till 72 hours after treatment with 1 micromol/L and 10 micromol/L ATRA. Forty-eight hours after exposed to 100 micromol/L ATRA, the enhancement of GJIC was less obvious. There was no significant increase induced by ATRA on the transcription of Cx43 gene, as demonstrated by semiquantitative RT-PCR.</p><p><b>CONCLUSION</b>ATRA turned out to be a potent enhancer on GJIC function in C6 glioma cells, andthe enhancement effect was most probable at post-transcriptional level.</p>