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Objective To investigate the effect of dipeptidyl peptidase-4 (DPP-4) inhibitor on lipopolysaccharide (LPS)-induced changes in the mass and function of pancreatic β-cells.Methods RINm cells were cultured and treated with LPS alone or combined with different concentrations of sitagliptin for 24 h.The proliferation of RINm cells was detected by CCK-8 assay.Apoptotic rate was determined by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide flow cytometry.Insulin secretion was measured by enzyme-linked immunosorbent assay.The expression of IL-6 mRNA was displayed by RT-PCR.Results LPS significantly stimulated the proliferation of RINm cells (0.89 ± 0.04 vs 1.14 ± 0.08,P＜0.01),while LPS+sitagliptin showed no significant difference compared with LPS group.The cell apoptotic rate in LPS + 10-1 mmol/L sitagliptin group was significantly lower than that in LPS group.There were no significant differences in basal insulin secretion among all groups,but after the high/low glucose stimulation,LPS increased insulin secretion as compared with the control.The IL-6 mRNA expression in LPS+sitagliptin group was significantly lower than that in LPS group (0.77 ± 0.33 vs 1.30 ± 0.41,P =0.006).Conclusions DPP-4 inhibitor has no influence on LPS-induced proliferation of pancreatic β-cell,but it can inhibit LPS-induced apoptosis and insulin secretion,and IL-6 may be involved in the process.
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ObjectiveTo investigate serum lipopolysaccharide (LPS) level in people with different glucose tolerances and to explore the relationship between LPS and insulin resistance/β-cell secretory function.Methods Sixty-seven subjects were recruited,including 23 with newly diagnosed type 2 diabetes ( T2DM),21 impaired glucose tolerance ( IGT),and 23 normal glucose tolerance (NGT).Serum LPS was assayed by limulus amebocyte lysate test ;expression of LPS toll-like receptor 4 (TLR4) on surface of plasma monocytes was measured by flow cytometric assays,and the changes of LPS levels by 0.5 hours and 2 hours after a high-fat diet were detected.Insulin resistance was evaluated by homeostasis model assessment for insulin resistance (HOMA-IR); β-cell secretory function was evaluated by homeostasis model assessment for β3 cell function ( HOMA-β )/HOMA-IR,increment in insulin in the first 30 minutes/increment in glucose in the first 30 minutes ( AIns30/ΔG30)/HOMA-IR,AUCIns120min/HOMA-IR.Results2 h LPS levels after a high-fat diet were significantly higher than fasting LPS levels [ NGT:0.96(0.33,0.99)vs 0.62 (0.22,0.64),IGT:1.08(0.53,1.22)vs 0.71 (0.39,0.82),T2DM:1.23 (0.62,1.43)vs 0.86( 0.45,0.94 ),EU/ml,all P＜0.01 ].Fasting,0.5 h,and 2 h LPS levels and fasting TLR4 levels of T2 DM group and IGT group were respectively higher than those of NGT group [ fasting LPS:0.86( 0.45,0.94 ),0.71 ( 0.39,0.82 ) vs 0.62(0.22,0.64),EU/ml;0.5 h LPS:1.10(0.55,1.18),0.84(0.50,1.07) vs 0.73(0.31,0.76),EU/ml;2 h LPS:1.23(0.62,1.43),1.08(0.53,1.22)vs 0.96(0.33,0.99),EU/ml; fasting TLR4:36.96( 17.22,55.19),30.34 ( 15.00,45.18 )vs 15.66 (6.09,9.76),MIF/105 cells,all P＜0.01 ].Fasting LPS,AUCLPS 120 min,and fasting TLR4 were positively correlated with insulin resistance index and negatively correlated with β-cell secretory function index ( P＜0.05 ).Multiple linear regression analysis showed that fasting LPS was an independent correlative factor of HOMA-IR and 0.5 h LPS was an independent correlative factor of (AIns30/AG30)/HOMA-IR and AUCIns Ins120min/HOMA-IR.ConclusionPeople with different glucose tolerances show differed LPS levels and its receptor TLR4 levcls,both of which are correlated with insulin resistance and β-cell secretory function,suggesting that LPS is associated with the pathogenesis of abnormal glucose regulation.
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To compare the effects of four different intensive insulin therapies on blood glucose control and vascular endothelial function in newly-diagnosed type 2 diabetes.Patients were randomly divided to accept pre-meal insulin aspart 30 or pre-meal insulin aspart and glargine at bedtime or pre-meal Novolin-R and NPH at bedtime or continuous subcutaneous insulin aspart infusion.Capillary blood glucose determination and continuous glucose monitoring system were carried out,therapeutic time and total insulin dosage were recorded.Ultrasound was used to evaluate the vascular endothelial function.Glucose level,incidence of low glucose,potency ratio of the four groups were similar( P＞0.05 ) ; FMD and NMD were not significantly improved ( P =0.718,P =0.065 ).The short-term efficacy and safety of the four groups are similar.The short-term intensive insulin therapy has no obvious effect on vascular endothelial function.
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Objective To investigate the the relationship of a high risk serum screen for Down syndrome in second trimester and adverse pregnancy outcomes,and to evaluate the predictive value for adverse pregnancy outcomes.Methods The tri-marker second trimester maternal serum screening for Down syndrome (alpha-fetoprotein,free beta-hCG and unconjugated estriol)was performed on the pregnant women at Peking Union Medical Hospital from January 2009 to January 2011.The cutoff valvue was 1/270.Pregnancy outcomes were followed up.The general condition and pregnancy complications of the pregnant women with high risk (high-risk group) were compared to that of the pregnant women with low risk (low-risk group); and with 35 years old as a demarcation,the incidences of adverse pregnancy outcomes were calculated in the two groups.Results ( 1 ) A total of 1935 cases were collected.And 1784 cases were with low risk,and 151 cases were with high risk.The difference of weight and gestational age betweem the two groups was not statistically significant ( P ＞ 0.05 ) ; the difference of age between the two groups was statistically significant ( P ＜ 0.01 ).(2) Pregnancy complications were found in 791 cases.In high-risk group,the incidences of gestational diaetes mellitus (GDM,13.9％),neonatal asphyxia (4.0％ ) and small for gestational age infant ( SGA,4.6％ ) were higher than that in low-risk group ( 8.4％,1.0％,1.6％ ),the difference was statistically significant ( P ＜ 0.05 ).The incidences of gestational hypertension disease,premature labor,oligohydrammios,placenta previa,placenta abruption,fetal macrosomia in the two groups was not statistically different (P ＞0.05).(3) In 1705 cases aged less than 35 years,129 cases (7.6％) were GDM,43 cases ( 2.5％ ) were gestational hypertension disease,61 cases ( 3.9％ ) were premature labor; in 230 cases aged 35 years or more,41 cases (17.8％ ) were GDM,12 cases (5.2％) were gestational hypertension disease,15 cases (6.5％ ) were premature labor,and the difference between the two groups was statistically significant ( P ＜ 0.05 ).In ＜ 35 years old group,the incidences of GDM,neonatal asphyxia and SGA (12.3％,4.4％,5.3％ ) were higher in the high-risk group than that (7.2％,0.9％,1.6％ ) in the low-risk group,and the difference was statistically significant ( P ＜ 0,05 ).In ≥35 years old group,the incidences of GDM,neonatal asphyxia and SGA ( 18.9％,2.7％,2.7％ ) were slightly higher in the high-risk group than that (17.6％,1.6％,1.6％ ) in the low-risk group,the difference between the two groups was not statistically significant (P ＞ 0.05 ).Conclusions The present study revealed apparertt increase in the adverse pregnancy outcomes in women with a high risk of Down syndrome screening test.Advanced age is the most important risk factor for a high risk of Down syndrome screening test and adverse pregnancy outcomes.More attention should be attached to the patients whose age were ＜35 years old and with a high risk of Down syndrome screening test.
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ObjectiveTo investigate the effects of lipopolysaccharide ( LPS ) on cell apoptosis,proliferation, and insulin secretion in a β-cell line, NIT-1. MethodsNIT-1 cells were stimulated with 1 μg/ml LPS for 0-120 h. Cell apoptosis was evaluated by Hochest33342 staining and Annexin V/PI flow cytometry. Cell proliferation was evaluated by CCK-8 and BrdU assay. Intracellular insulin content, basal insulin secretion, and glucose-stimulated insulin secretion(GSIS) were detected by RIA. The IRS-2 tyrosine phosphorylation was determined by Western blot. ResultsCell apoptosis was not significantly changed by treatment with LPS for 120 h. Cell proliferation was stimulated by LPS before 48 h, and inhibited after 96 h. Intracellular insulin content or GSIS was not altered, but basal insulin secretion was decreased significantly by LPS after 48 h ( all P＜0.01 ). LPS decreased the tyrosine phosphorylation level of IRS-2 ( 0. 45 ± 0. 08 vs 0. 22 ± 0. 06, P ＜ 0. 05 ) and stimulated IκBα phosphorylation. Pretreatment with a specific IκBα phosphorylation inhibitor, Bay1 1-7082 for 1 h, remarkably blunted the LPS-induced phosphorylation of IκBα and cell proliferation( both P＜0.01 ). ConclusionsLow-dosages of LPS regulate proliferation and basal insulin secretion of NIT-1 β-cells, in which activation of NF-κB and inhibition of IRS2 tyrosine-phosphorylation may be involved.
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Objective To analyze the relationship between karyotypes and clinic features of patients with primary amenorrhea. Method Karyotype analysis of patients with primary amenorrhea was performed by using G-banding technique. Results Karyotype analysis of 468 patients with primary amenorrhea revealed that 255 patients (54. 49% ) had normal female karyotypes and 213 patients (45.51%) had abnormal karyotypes, including 143 patients with abnormal X chromosome, 4 patients with mosaic X -Y chromosome, 57 patients with 46, XY karyotype, 8 patients with abnormal autosome and one patient with Xautosome translocation. 75.52% primary amenorrhea patients with short stature had abnormal X chromosome, and all primary amenorrhea patients with deletion or break-up of Xp11. 1 - 11.4 and Xp21 - 22 were short statures. Conclusion One of the main reasons of primary amenorrhea was chromosome abnormity,especial heterosome abnormity. Karyotype analysis should be used to detect primary amenorrhea patients in regular. There might be relationship between height improvement and the abnormity of Xp11. 1 - 11.4 and Xp21 - 22.
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AIM:To investigate the production and activation of caspase-3 in primary rat renal proximal tubule cells in response to tumor necrosis factor-α(TNF-α) and the implication of nuclear factor-κB (NF-κB) in the process. METHODS:Isolated rat renal proximal tubule cells (PTCs) from male adult Sprague Dawley rats were treated with TNF-α according to the indicated time courses. A specific NF-κB inhibitor,Bay11-7082,was used alone or as a pretreatment for 1 h followed by exposure to TNF-α for 24 h.The protein levels of cleaved caspase-3,caspase-3,I-κBα,phosphorylated I-κBα,and GAPDH were detected by Western blotting using specific antibodies. RESULTS:The protein level of cleaved caspase-3 relative to caspase-3 was significantly increased in the presence of TNF-α for 6 h,12 h,and 24 h. Protein levels of caspase-3 were significantly decreased by 12 h and returned to baseline by 24 h in the presence of TNF-α. Treatment with Bay11-7082 for 25 h alone or pretreatment with Bay11-7082 for 1 h followed by addition of TNF-α for 24 h caused a remarkable reduction in both cleaved caspase-3 and caspase-3 as compared to control and TNF-α treated groups. An increase in phosphorylated I-κBα was observed from 15 min to 60 min after treatment with TNF-α at a dose of 10 μg/L in PTCs. CONCLUSION:NF-κB is not only associated with the activation of caspase-3 but also the production of caspase-3 in primary rat renal proximal tubule cells in response to TNF-α.
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Objective To investigate the effect of miR-375 inhibited by 2'-O-me-375 on lipoapoptosis of NIT-1 pancreatic β cells. Methods NIT-1 cells were divided and treated according to the optimal condition: mock (without lipofectamine) ,lipofectamine( transfected only with lipofectamine) ,NC-miRNA (transfected with negative control miRNA) ,and 2'-O-me-375( transfected with 2'-O-me-375) groups. 72 hours later, all cells in each group were cultured with 500 μmol/L palmitate for 48 h. The percentage of apoptotic cells was detected by Hochest33342 staining and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL). The protein expression of myotrophin ( V1 ) , a target gene of miR-375, was detected by Western blotting. Results Compared to the other three groups,the cell apoptosis rate of 2'-O-me-375 group was the lowest (P<0.01) .along with the highest VI expression level(P<0. 01). Conclusion Inhibition of miR-375 decreases pancreatic (3-cell lipoapoptosis.
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05). Conclusion The maternal serum level of ADAM 12 in the first-trimester is a potential marker for aneupolyhaploid screening and early fetal loss prediction, and is suggested to be tested at 9-12 gestational weeks as part of prenatal screening.
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Objective To evaluate the performance characteristics of the second trimester double-marker test for the detection of fetal Down's syndrome in mainland China. Methods This prospective national multi-centered study used alpha-fetoprotein (AFP) and free β-subunit of human chorionic gonadotrophin( free β-hCG)as the serum markers. From May 2004 to September 2006, 11 centers participated in the collection and analysis of maternal serum AFP and free β-hCG between 14 and 20+6 weeks of pregnancy. The screening results were calculated using the standard algorithm based on the standard database provided with the analytic software. Patients with an increased risk of Down's syndrome pregnancy (≥1/270) were offered genetic anmiocentesis. Outcomes of all pregnancies were obtained.Results A total of 66 132 singleton pregnancies were included in the study. The median maternal age was 27 years. At a cut-eft of 1 in 270, the detection rate (DR) based on a Caucasian database was 72% corresponding to a false positive rate (FPR) of 5%, and the DR based on the Chinese database was raised to 76% corresponding to an FPR of 5%. Conclusion The double-marker test using AFP and free β-hCG is an effective screen strategy for second-trimester detection of fetal Down's syndrome in mainland China. Ethnic variance exists between the Caucasian and Chinese populations. The accuracy of screening is increased by the use of race-specific medians.
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Objective To compare the efficacy and safety of three regiments of transient intensive insulin therapy for type 2 diabetes mellitns: thrice preprandial injection of premixed insulin aspart 30, thrice preprandial injection of insulin aspart and injection of glargine at bedtime, thrice preprandial injection of regular insulin and injection of NPH at bedtime. Methods Patients were randomly divided into 3 groups, treated with 3 kinds of intensive insulin therapy. After achieving the target goal, continuous glucose monitoring system was used to compare the blood glucose level, therapeutic time, dosage of insulin, occurrence of hypoglycemia. Results Detected by continuous glucose monitoring system, there was no statistical difference in average blood glucose [(8.3±2.1,7.5±1.9, 6.8±0.8) mmol/L, P > 0.05], blood glucose area under curve 3 hours (AUC1-3) after breakfast, therapeutic time [ (8.3±2.5, 9.1±3.8, 8.4±1.7)d, P > 0.05], dosage of insulin [(0.63± 80%, P > 0.05) among three kinds of transient intensive insulin therapy. There were no patients complaining of hypoglycemic symptom. Conclusion The short-term efficacy and safety among three intensive insulin therapeutic methods are similar. More attention should be paid to monitor the blood glucose during sleep.
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0.05,respectively).Plasma Mg2+,intracellular Mg2+,the beta 2-AR mRNA and protein in lung tissue in group C at 21st d and 34th d were significantly higher than those in group A at 21st d and 34th d 21st d:(0.84?0.09)mmol/L vs 0.57?0.10)mmol/L,(2.39?0.14)mmol/L vs(2.11?0.08)mmol/L,(0.75?0.09)pmol/g vs(0.59?0.06)pmol/g,(88.50?8.50)pmol/g vs(60.10?7.70)pmol/g,P
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AIM: To investigate the immune stimulation capacity of B7-H1 blockade on immature dendritic cells (DCs) in vitro. METHODS: The human monocyte-derived dendritic cells were induced in the presence of cytokine GM-CSF and IL-4. The expression of B7-H1 was detected by FCM. On blockade of B7-H1, the maturation and endocytic activity, T cells stimulatory proliferation capacity, IL-12 production, T cell differentiation effect of DCs were detected by FCM, MTT assay, ELISA and ELISPOT, respectively. RESULTS: The expression of B7-H1 was increased with the induction of DCs. On day 7, the positive expression was 54.12%, and the TNF-? induced mature DCs had the positive expression rate of 83.64%. The blockade of B7-H1 on immature DCs had sharply increased their T cells stimulatory proliferation capacity and IL-12 production, and efficiently induced the development of Th1/Tc1 cells, but had no effect on their maturation and endocytic activity. CONCLUSION: The blockade of B7-H1 on immature DCs increases its immune stimulation activity. It is valuable to investigate the antitumor immune responses of DCs vaccine with B7-H1 blockade.
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Aim To investigate the effect of insulin glargine and human insulin on proliferation of a human breast cancer cell line MDA-MB-231 and the role of ERK in the process.Methods MDA-MB-231 cells were incubated with insulin glargine and human insulin at different concentrations and for different time courses.A specific ERK1/2 inhibitor,PD98059,was used either alone or in combination with insulin glargine or human insulin to test the involvement of ERK pathway in cell growth.Cell proliferation was evaluated using cell counting kit-8 reagents.Cell cycle distribution was analyzed by flow cytometry.Results Both insulin glargine and human insulin dose-dependently enhanced MDA-MB-231 cell proliferation at the concentrations from 1 to 100 IU?L-1 after treatment for 96 h.At the concentration of 10 IU?L-1,both drugs promoted cell growth at 48,72,and 96 h.The percentage of S+G2/M cells was significantly increased in both insulin glargine and human insulin treated groups as compared to untreated controls.No significant difference was observed between insulin glargine and human insulin in their effects on cell proliferation and cell cycle distribution.Cell proliferation was significantly inhibited by PD98059.However,in the presence of PD98059,both drugs still promoted cell proliferation significantly as compared to untreated controls.Conclusions Insulin galrgine and human insulin similarly promote proliferation of MDA-MB-231 cells independent of ERK activation.