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Objective To compare ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) and chemiluminescence immunoassay(CLIA) measurement of human serum androgen levels in polycystic ovarian syndrome(PCOS). Methods 160 PCOS patients and 146 healthy subjects(control group) were recruited and their serum androgen levels——measurements of testosterone and dehydcoepiandrosterone sulfate (DHEA-S) were tested by UPLC-MS/MS and CLIA. The androgen results combined with body mass index(BMI) were analyzed by ROC curve. According to area under curve(AUC) calculated by SPSS, a better method will be selected to provide accurate test results for physicians. Results (1)AUC of DHEA-S tested by UPLC-MS/MS was significantly higher than the one tested by CLIA(P<0.01). There was no significant difference between the AUC of T tested by UPLC-MS/MS and the one tested by CLIA. (2)AUC of T combined with DHEA-S tested by HPLC was not only significantly higher than the AUC of two combined indicators tested by CLIA(P<0.01),but also significantly higher than the AUC of a single indicator——either T(P<0.01) or DHEA-S(P<0.01) tested by UPLC-MS/MS. (3)The AUC of T,DHEA-S combined with BMI tested by HPLC was not only significantly higher than the AUC of three combined indicators tested by CLIA(P<0.01),but also higher than the AUC of two combined indicators tested by UPLC-MS/MS(P<0.05). Conclusion T and DHEA-S tested by UPLC-MS/MS combined with BMI has a certain reference value in the diagnosis of PCOS.
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Objective To explore the effect of nicotinaide nucleotide transhydrogenase(NNT) mutation on glucose homeostasis in C57BL/6 mice with mix background. Methods We generated wild type NNT homozygous, mutant NNT homozygous and heterozygous by mating the C57BL/6J (with NNT mutation) and 6N (without NNT mutation). At the age of 4 weeks, those mice were randomly assigned to normal control diet(NCD) or high-fat diet(HFD) for 4 weeks. The body weight was measured every week. At the age of 8 weeks, an intraperitoneal glucose tolerance test(IPGTT) and an intraperitoneal insulin tolerance test (ITT) were performed. Results The body weight growth was not affected by NNT mutation during an HFD fed. NNT mutant mice showed significant glucose intolerance. After 4 weeks of high fat diet, the NNT mutant mice showed a decreased insulin sensitivity, while the glucose excursion curve was not elevated in the heterozygous mice. Conclusion NNT mutation had a significant influence on the phenotype of glucose metabolism and insulin resistance of mice, in particular under a metabolic stress. The phenotypes of heterozygous and homozygous mutant ones differed from each other. When using mice with C57BL/6J and C57BL/6N mixed background in research, NNT mutation should be carefully screened in all metabolic studies.
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Objective · To explore the relationship between baseline lipid profiles and long-term cardiovascular outcomes after intervention with hypoglycemic drugs metformin and glipizide and to detect lipid components that can predict the long-term cardiovascular effect of metformin and glipizide.Methods· Liquid chromatography-quadrupole time of flight-mass spectrometry (LC-QTOF/MS) was used to measure 119 lipid components in baseline serum for 116 patients with type 2 diabetes (T2DM) and atherosclerotic heart disease (CHD) who were treated with glipizide (56 cases,the glipizide group)or metformin (60 cases,the metformin group).Cardiovascular complex end points (including cardiovascular death,all-cause death,nonfatal myocardial infarction,nonfatal stroke,and arterial revascularization) of all patients were followed up.The relationship between lipid components and cardiovascular complex end points was analyzed with Logistic regression analysis.The category-free net reclassification index (cfNRI) and the integrated discrimination improvement (IDI) were used to evaluate whether lipid components are helpful for predicting the recurrent cardiovascular events.Results· The differences in baseline drug distribution,clinical characteristics,and biochemical indexes between two groups were not statistically significant,except for diuretics use,serum PC (O-34:2) level,and SM (d18:0-24:0) level.Logistic regression analysis showed that baseline ChE (20:4) was a protective factor for recurrent cardiovascular events in the glipizide group (OR=0.87,P=0.039).ChE (20:4) significantly increased the cfNRI and IDI of cardiovascular complex end points by 69％ and 0.07,respectively (P=0.011,P=0.028).Baseline SM (dl 8:1-22:0) was a risk factor for recurrent cardiovascular events in the metformin group and all participants (OR=1.65,P=0.039;OR=1.64,P=0.014).SM (d18:1-22:0) significantly increased the cfNRI of cardiovascular complex end points in the metformin group and all participants by 74％ and 55％,respectively (P=0.012,P=0.005).Conclusion· Of 119 lipid components measured with LC-QTOF/MS,baseline ChE (20:4) is a protective factor and SM (d18:1-22:0) is a risk factor for cardiovascular complex end points in with T2DM and CHD patients after long-term treatment with metformin and glipizide.Both lipid components are helpful for improving the prediction of recurrent cardiovascular events.
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mTOR pathway plays a critical role in cell proliferation, growth and metabolism. This pathway is composed of two different large protein complexes, mTORC1 and mTORC2, which have their distinct downstream effects. Its inhibitor, rapamycin, has been proved to cause β-cell damage and glucose intolerance. Furthermore, various transgenic mouse models and ex vivo studies have revealed that mTORC1 and mTORC2 are both essential for maintaining normal β cell mass and function, whereas the underlying molecular mechanism and the relevance of the whole mTOR signaling to pathogenesis of type 2 diabetes remain to be explored and further clarified.
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Objective To investigate the adrenal steroidogenic function in genotype-proven heterozygotes carrying mutations in CYP17A1 gene in vivo. Methods Eight patients and 14 family members from 5 families with 17-hydroxylase/17,20-lyase deficiency (17OHD) were recruited. The mutations of the CYP17A1 gene in these individuals were screened by direct sequencing of PCR products. The hormonal response to ACTH was evaluated in the 14 genotype-proven carriers and 45 age- and sex-matched normal subjects. Results Three mutations were found in 5 unrelated families. 14 carriers with CYP17A1 mutation were identified, including 7 heterozygotes with D487_F489del, 6 with Y329fs, and 1 for H373L. Compared to the normal subjects, the carriers exhibited lower basal and ACTH-stimulated cortisol levels, but higher ACTH-stimulated corticosterone level. The ratios of corticosterone to cortisol in the genotype-proven heterozygotes were higher than those of normal individuals at baseline and following ACTH-stimulation. Similarly, progesterone level and ratios of progesterone to 17-hydroxyprogesterone in the male heterozygotes were also higher than that of normal individuals before and after stimulation. No significant differences were observed in the hormone levels between two genotypes (D487_F489del vs Y329fs). Conclusions Genotype-proven carriers of 17OHD without apparent clinical symptoms exhibit decreased enzyme activity,analogous to mildly impaired adrenal 21-hydroxylase activity in the carriers of CYP21 A2 gene mutation.
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Objective Cathepsin K (CTSK) played an important role in adipocyte differentiation.The activation of CTSK needs to convey by mannose-6-phosphate receptors (M6PR) in osteoclasts. The aim of the present study was to identify the effects of mannose-6-phosphate (M6P) in adipocyte differentiation and its underlying molecular mechanism. Methods Oil red O staining, accumulation of cytoplasmic triglycerides and glycerine release were used to assess its effects on adipocyte differentiation in the 3T3-L1cell line. The enzyme activity of CTSK was observed by laser confocal microscopy. The proliferation of 3T3-L1 preadipocytes was detected by MTT methods. mRNA expression of M6PR was determined by RTPCR. Results M6P could prevent adipocyte differentiation in a dose-dependent manner as evidenced by absence of triglyceride accumulation and glycerol content. Statistical significance was showed when the concentrations of M6P were 5.0 mmol/L and 8. 0 mmol/L respectively(P ＜0. 05). The mRNA expression of M6PR was detected during the whole process of adipocyte differentiation. With the increase of M6Pconcentration, enzyme activity of CTSK was inhibited in a concentration-dependent manner. MTT method showed that the absorbance at 570 nm of 3T3-L1 preadipocytes was 0. 057 ±0. 091, increased about 62. 9%at 10. 0 mmoL/L compared with the control group (P ＜ 0. 05 ). Conclusion M6P inhibits the terminal differentiation of adipocyte, which may be associated with its effect of blocking CTSK activity by competitive binding with M6PR.
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Objective This research attempted at prevention of steam contamination and protection of medical staff in periodontal scaling. Methods Thirty sample patients without oral cavity or upper res-piratory tract infection, chronic disease, infectious disease or other contraindication were randomly classified into the experimental group and the control group.The former gargled with chlorhexidine in the process of periodontal scaling while the latter rinsing only. Oral cavity bacteria of both groups were sampled before and after operation;air samples were got at five periods of time(before operation, 10 minutes in operation, 20 minutes in operation, 30 minutes in operation, and end of operation) and germicnltured.The entire data were analyzed.Results Air samphng in periodental scaling and oral sample germiculture after the scaling in the experimental group were lower than those of the control group, P<0.01. Conclusions With effective disinfection and sta-ble characteristics, chlorbexidine can be popularly utilized in skin mucous membrane disinfection. Gargling with chlorhexidine in periodontal scaling eliminates bacteria and microorganism in oral cavity and resulting steam,thus reducing environmental pollution and protecting the health of medical staff.
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Betacellulin (BTC) is one of "islets regeneration factors" which received more and more attention these years. BTCe is the C-terminal 50-residue region of mature BTC protein and can bind with erbB-1?erbB-4 receptor. It has the same mitogenic activity on cells as the whole section. BTC and BTCe can improve the level of glucose-stimulated insulin secretion (GSIS) during islets culture in vitro though they had no effect on the acute insulin secretion. BTCe also effectively ameliorated the hyperglycemia of STZ-induced diabetic rats by a single plasmid injection into muscle of rats. It is supposed that BTCe promote the proliferation of PDX-1 positive cells or repair some signal transduction pathway. Perhaps the latter is more important.
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Objective To explore the possible correlation between serum highly sensitive C reactive protein (hs CRP) and blood glucose, insulin, lipids ,insulin sensitivity index (SI),acute insulin response(AIR), and adiponectin in subjects with impaired glucose tolerance (IGT) or newly diagnosed type 2 diabetes mellitus(DM). Methods SI and AIR were assessed by the reduced sample number of Bergman′s minimal model method by intravenous glucose tolerance test in subjects. Meanwhile body mass index (BMI), waist hip ratio (WHR), the serum lipid profile, hs CRP, adiponectin levels were measured. Results Compared with normal control (NC) group[SI(6.6?2.4) 10 -4 (min?mU/L) -1 ,adiponectin7.77(6.35 10.70 mg/L),hs CRP0.40(0.21 1.67mg/L)], the SI and serum adiponectin in IGT group [(1.5?1.1) 10 -4 (min?mU/L) -1 , 4.29(3.59 6.22 mg/L) respectively] and type 2 DM group [(1.5?1.0)?10 -4 (min?mU/L) -1 , 3.46(2.37 4.72 mg/L) respectively] were significantly decreased (all P
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Objective To study the association of glucagon like peptide 1 receptor (GLP1R) gene polymorphism with type 2 diabetes in Han population in Shanghai. Methods In the study, 360 type 2 diabetic patients and 313 normal control subjects were enrolled. Diabetic patients were further subdivided into insulin treated non obese patients (BMI28, 192 subjects). A single nucleotide polymorphism (SNP) rs 2268657 was genotyped in all the subjects enrolled in the study using allele specific real time PCR and its association with type 2 diabetes was examined. Results The frequencies of AA,AG, GG genotype incontrol group were0.086,0.447, 0.466 respectively, 0.155, 0.375, 0.470 in non obese diabetic patient group respectively, and 0.109, 0.500, 0.391 in obese diabetic patient group respectively. There was significant difference of the frequency of genotype AA between control group and non obese diabetic patient group (OR=1.939, P
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The relationship between abnormal circulating GH-insulin-like growth factor (IGF) axis and hypoglycaemia was explored in 2 cases of non-islet cell tumor associated hypoglycaemia (NICTH). Serum level of IGF-Ⅱ was increased, the levels of GH, IGF-Ⅰ, IGFBP-3 were decreased to some extent, and IGF-Ⅱ/IGF-Ⅰ (molar ratio) was significantly increased in these two cases. In one case, circulating GH-IGF axis returned to normal after complete removal of the tumor. In the diagnosis of NICTH, the abnormal circulating GH-IGF axis is quite valuable and IGF-Ⅱ/IGF-Ⅰ appears to be a more sensitive parameter.