Association of adiponectin gene polymorphism with obesity in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the distribution characteristics of adiponectin gene +45 single nucleotide polymorphisms (SNP) in Chinese children, and to determine the role of adiponectin gene +45 polymorphisms in the pathogenesis of childhood obesity.</p><p><b>METHODS</b>A total of 147 Chinese obese and 118 healthy children were randomly selected and enrolled to identify adiponectin gene SNP+45 polymorphism by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. Plasma adiponectin levels were determined using ELISA. Waist circumference (WC), waist to hip ratio (WHR), percentage of body fat (%BF), systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting plasma glucose (FPG), serum triglycerides (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), plasma fasting insulin (FINS), and homeostasis model assessment of insulin resistance (HOMA-IR) were measured.</p><p><b>RESULTS</b>The allelic frequency of adiponectin gene SNP+45 in children with obesity and healthy controls were 40.5% and 25.4%, respectively. There were significant differences in the distribution of genotypes and the allelic frequency between the two groups (P<0.05). The plasma adiponectin levels were significantly higher, in contrast, %BF, HOMA-IR, TC and LDL-C levels were significantly lower in obese children with TT genotype than those in obese children with TG or GG genotype.</p><p><b>CONCLUSIONS</b>The adiponectin gene SNP+45 polymorphism may be associated with pathogenesis of obesity in children. T→G variance may be associated an increased risk of childhood obesity and result in a decreased level of adiponectin.</p>

Effect of repaglinide and gliclazide on glycaemic control, early-phase insulin secretion and lipid profiles in / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Both repaglinide and gliclazide are insulin secretagogues widely used in the treatment of type 2 diabetes. They stimulate insulin secretion through distinct mechanisms and may benefit patients from different aspects. The present study was to evaluate the effects of repaglinide or gliclazide on glycaemic control, insulin secretion, and lipid profiles in type 2 diabetes patients.</p><p><b>METHODS</b>A total of 47 newly diagnosed type 2 diabetes patients were randomized 1:1 to receive a 4-week treatment with repaglinide or gliclazide. The standard mixed meal tolerance test was performed before and after the treatment. Plasma glucose (PG), insulin concentration, and lipid profiles were measured. The area under insulin concentration curve (AUC(ins)) and the early-phase insulin secretion index (ΔI(30)/ΔG(30)) were calculated.</p><p><b>RESULTS</b>After the trial, fasting and postprandial PG and postprandial insulin improved significantly in both groups (P < 0.05). The maximum insulin concentration occurred earlier in the repaglinide group than that in the gliclazide group. AUC(ins) increased in both groups (P < 0.05), but no significant difference was found between groups. ΔI(30)/ΔG(30) increased in both groups (P < 0.05), especially in the repaglinide group (P < 0.05). Triglyceride and total cholesterol decreased significantly in the repaglinide group in some time points, while no significant change was observed in the gliclazide group.</p><p><b>CONCLUSIONS</b>Repaglinide and gliclazide had similar effects on glycaemic control and total insulin secretion, while repaglinide had more effects on improvements in β-cell function and lipid metabolism.</p>

Expression of imprinted genes during the course of differentiation from mouse embryonic stem cells to islet-like cells in vitro / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of in vitro inducement on the expression of SF1-G imprinted genes, Kcnq1 and Cdkn1c during the course of differentiation from mouse embryonic stem (ES) cells to islet-like cells.</p><p><b>METHODS</b>Mouse ES cells were induced to differentiate into islet-like cells in vitro. The expression of islet specific markers was tested by RT-PCR or immunofluorescence. RT-PCR/RFLP was used to test the imprinted genes parental expression in cells at different stages.</p><p><b>RESULTS</b>Islet specific genes, such as Insulin, Glucagon, Somatostatin, IAPP and Glut2, were expressed in differentiated cells. The proteins of insulin, C-peptide and Somastatin were expressed in the final stage cells. Imprinted gene Kcnq1 and Cdkn1c were biallelicly expressed in islet-like cells.</p><p><b>CONCLUSIONS</b>Mouse ES cells can be successfully induced into islet-like cells in vitro. Gene imprinting status of Kcnq1 and Cdkn1c may be changed in differentiated cells (causing loss of imprinting) during the in vitro inducement.</p>

Serum leptin level and its association with bone mineral density in obese children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate serum leptin level and its relationship with bone mineral density in obese children from Changsha City.</p><p><b>METHODS</b>One hundred and nineteen obese children and 103 normal children aged 7 to 12 years from five primary schools of Changsha City were enrolled. Obesity was assessed based on the body mass index (BMI). Dual energy X-ray absorptiometry (DEXA) was used to determine bone mineral density (BMD) and body composition. Serum leptin level was measured using enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The obesity group had higher height, weight, BMI, waist circumference and waist to hip ratio (WHR) compared with the normal group (p<0.01). BMD, bone mineral content (BMC), lean mass (LM), fat mass (FM), percentage of body fat (%BF) and leptin concentration in the obesity group were significantly higher than those in the normal group (p<0.01). Serum leptin level was positively correlated with BMD, BMC, LM and FM (r=0.528-0.903, p<0.01). Multiple stepwise regression analysis indicated that BMI and %BF were independent influencing factors for serum leptin level.</p><p><b>CONCLUSIONS</b>Obese children have higher serum leptin level. Serum leptin concentration is significantly correlated with BMD and body composition. BMI and %BF are independent influencing factors for serum leptin level in children.</p>

Hyperlipidemia induced by high fat diet ingestion activates TGF-beta/Smad signaling pathway in the kidney of diabetic rats / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of diet-induced hyperlipidemia on TGF-beta/Smad signaling pathway in the kidney of diabetic rats, and to explore the mechanism by which hyperlipidemia leads to renal injury in diabetes.@*METHODS@#Diabetic rats and non-diabetic rats were fed with normal fat diet and high fat diet for 16 weeks, respectively. The expressions of TGF-beta1, TbetaRII, and Col-IV mRNA in the renal cortex were examined by reverse transcriptase-PCR,TbetaRII and p-Smad staining in glomerular cells were detected by immunohistochemical staining, and the expression of TGF-beta1 and Col-IV protein was determined by Western blot.@*RESULTS@#Diet-induced hyperlipidemia up-regulated the levels of TGF-beta1, TbetaRII, p-Smad, and Col-IV protein and mRNA in the renal cortex of diabetic rats compared with those of non-diabetic rats. However, feeding high fat diet to non-diabetic rats had no influence on the expression of TGF-beta1, TbetaRII, p-Smad2, and Col-IV in the renal cortex.@*CONCLUSION@#Hyperlipidemia induced by high fat diet ingestion leads to renal injury in diabetic rats through activating TGF-beta1 /Smad signaling pathway.

Changes of endothelium-dependent vasodilation in patients with impaired glucose tolerance and type 2 diabetes / 中南大学学报(医学版)

OBJECTIVE@#To investigate the changes of endothelium-dependent flow-mediated dilation (FMD) in patients with impaired glucose tolerance (IGT) and Type 2 diabetes (T2DM) and its influencing factors.@*METHODS@#The patients with IGT and T2DM were divided into IGT group (n=36), T2DM without vascular complication group (DM1;n=57), and T2DM with vascular complication group (DM2;n=31). And 25 normal subjects served as controls (NC group). The FMD was measured by high resolution ultrasound.The serum levels of tumor necrosis factor-alpha (TNF-alpha) and high sensitive C-reactive protein (hs-CRP) were detected with ELISA, and nitric oxide (NO) with Griess Reaction. The serum glucose, lipids, and other indexes were also detected.@*RESULTS@#Compared with the NC group, the serum levels of triglyceride (TG) total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), hs-CRP, and TNF-alpha significantly increased (P<0.05), but the serum levels of high density lipoprotein-cholesterol (HDL-C), the NO, and FMD significantly decreased (P<0.05) in the IGT, DM1, and DM2 group. With the progress of diabetes, FMD level descended. Pearson correlation analysis showed a negative correlation between FMD and glycosylated hemoglobin (HbA1c), insulin resistant index (HOMA-IR),TG,TC,hs-CRP, and TNF-alpha (P<0.01), but a positive correlation between FMD and NO,HDL-C (P<0.01) in IGT and T2DM patients. In multiple linear stepwise regression with FMD as dependent variable, NO,HbA(1)c,HDL-C,HOMA-IR, TNF-alpha, and hs-CRP showed a significant association with FMD (P<0.01).@*CONCLUSION@#Endothelium-dependent vasodilation is impaired in patients with IGT and T2DM,which is associated with hyperglycaemia, insulin-resistance, hyperlipemia, and inflammation.

Effect of rosiglitazone on NO and eNOS via PI3K/PKB signal pathways in cultured human umbilical vein endothelial cells / 中南大学学报(医学版)

OBJECTIVE@#To observe the effect of rosiglitazone on the production of nitric oxide (NO) and the expression of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB) /the endothelial nitric oxide synthase (eNOS) in cultured human umbilical vein endothelial cells(HUVECs), and to investigate the mechanism of signal transduction of rosiglitazone in improving the endothelial function.@*METHODS@#HUVECs were treated with various concentrations of rosiglitazone. The NO level was measured using Griess Reaction in cell culture supernatants; the expressions of PI3K-, PKB- and eNOS mRNA were measured using RT-PCR; and the expressions of PKB, eNOS, and phosphorylation of PKB-Ser473, eNOS-Ser1177 were measured using Western Blot.@*RESULTS@#Rosiglitazone increased the endothelial NO production in a dose- and time-dependent manner in cultured HUVECs, and also increased the expression of PI3K mRNA and the phosphorylation of PKB-Ser473 and eNOS-Ser1177 in a concentration-dependent manner, with no alteration in the expression of PKB and eNOS in cultured HUVECs. N(w)-nitro-L- arginine methyl ester (L-NAME, eNOS synthase inhibitor) blocked the rosiglitazone-induced NO formation; LY294002 (a PI3K inhibitor) prevented the NO production; and the phosphorylation of eNOS and PKB was induced by rosiglitazone.@*CONCLUSION@#Treatment with rosiglitazone can increase the NO production and improve the endothelial function through up-regulating the PI3K/PKB/eNOS signal pathways in cultured HUVECs.

Aortic endothelium-dependent vasodilation function and PI3K-, PKB-, eNOS mRNA expressions in insulin-resistant and type 2 diabetic rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To observe the changes of aortic endothelium-dependent vasodilation function (EDVR) and expressions of endothelial nitric oxide synthase (eNOS), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (PKB) in insulin-resistance (IR) and type 2 diabetic rats.</p><p><b>METHODS</b>IR rat model was established by feeding 4-6 week-old male SD rats with high glucose and cholesterol diet for 6 weeks and type 2 diabetes (DM) were induced by intraperitoneal injection with low dose streptozotocin (STZ) to IR rats. Glucose infusion rate (GIR) was determined by euglycemic hyperinsulinemic clamp technique, EDVR by acetylcholine (Ach)-induced vasodilation response in isolated aortic rings, aortic NO concentration by Griess Reaction, activation of eNOS detected by immunohistochemical SP method, mRNA expressions of eNOS-, PI3K- and PKB of aorta were assayed by RT-PCR, aorta ultrastructure observed by electron microscopy. Body weight, fast plasma glucose (FPG), insulin (FINS), triglyceride (TG), cholesterol (TC) were determined and the insulin sensitivity index (ISI) was calculated.</p><p><b>RESULTS</b>(1) Body weight, FINS, TG and TC levels were significantly higher while ISI and GIR significantly lower in IR and DM rats than that in normal control rats (P < 0.05). (2) Aorta EDVR decreased significantly in IR and DM group compared with that in control group (P < 0.05) and EDVR was also significantly reduced in DM rats than that in IR rats (P < 0.05). The maximum Ach-induced vasodilation response (EDVR(max), P < 0.01) was positively correlated with ISI and negatively correlated with FPG, TG, TC and FINS (P < 0.01). (3) Aortic NO concentration, the mRNA expressions of eNOS-, PI3K-, and PKB and eNOS immunohistochemical expression in aorta were significantly lower in IR and DM rats compared with normal control rats and the decrease was more pronounced in DM rats (P < 0.05 vs. IR). (4) Pathologic aortic ultrastructure changes were also visualized in IR and DM rats.</p><p><b>CONCLUSION</b>Our results suggest that reduced NO concentration and expression as well as reduced PI3K-, PKB-, and eNOS mRNA expressions might contributed to the reduced EDVR function and related pathological ultrastructure changes in IR and DM rats.</p>

Effects of rosiglitazone on endothelium-dependent vasodilation in patients with Type 2 diabetes / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effects of rosiglitazone on endothelium-dependent vasodilation in patients with Type 2 diabetes.@*METHODS@#Eighty-three newly diagnosed patients with Type 2 diabetes were divided into metformin group (metformin 750 mg/d), therapeutic alliance group (rosiglitazone 4 mg/d and metformin 750 mg/d), and rosiglitazone group (rosiglitazone 4 mg/d), and 25 normal subjects were as the control group. All patients were treated for 12 weeks. The height, weight, blood pressure, fasting plasma glucose (FPG), fasting serum insulin (FINS), glycosylated hemoglobin (HbA1c), triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), and serum nitric oxide (NO) were measured before and after the therapy. The body mass index (BMI) and homeostasis model assessment-insulin resistant index (HOMA-IR) were calculated. The flow-mediated dilatation (FMD) and endothelium independent dilatation (EID) were measured by high-resolution ultrasound.@*RESULTS@#The BMI, TG, TC, LDL-C, FPG, HBA1c, HOMA-IR were higher (P<0.01), but HDL-C, NO, FMD were lower (P<0.01) in all diabetic groups than those in the control group before the treatment. After the 12-week treatment, TG, FPG, HBA1c, HOMA-IR decreased significantly in all the diabetic groups (P<0.01); FINS decreased, but HDL-C, NO, FMD and EID increased (P<0.01) in therapeutic alliance group and rosiglitazone groups, while HDL-C, FINS, NO, FMD had no change in metformin group compared with those before the treatment.@*CONCLUSION@#Endothelium-dependent vasodilation function is impaired in patients with Type 2 diabetes. The therapy with rosiglitazone can increase the serum NO level, and improve endothelium-dependent vasodilation,but metformin has no influence on the above-mentioned indexes.

Effect of plasma glucose on the vascular endothelial function and analysis of relevant factors / 中南大学学报(医学版)

OBJECTIVE@#To compare the flow-mediated dilatation (FMD) among the newly diagnosed impaired glucose tolerance (IGT), type 2 diabetes mellitus (T2DM), and the normal controls (NC) and to analyze relevant factors under different glucose levels.@*METHODS@#The study included IGT (n=34), DM1 (n=52), DM2 (n=33) and NC (n=25). Levels of fasting plasma glucose (FPG), 2-hour postprandial plasma glucose (PPG), fasting insulin (FINS), 2-hour postprandial insulin (PINS), triglyceride (TG), total cholesterol (TC), and hemoglobin A1C (HbA1C) were determined in all participants. High resolution ultrasound examining FMD was performed to measure vascular endothelial function subsequently.@*RESULTS@#There was statistically significant difference between IGT, DM, HG and NC group in FMD (P=0.008). Partial correlation analysis found that a significant negative correlation existed between FMD and homeostasis model assessment-index (HOMA-IRI), difference of plasma glucose (DPG), FPG and PPG (P<0.01), and a negative correlation between FMD and HbA1C (P<0.05). Setting FMD as dependent variable to conduct multiple linear stepwise regression, in IGT group it was the waist/hip ratio (WHR) and HOMA-IRI that entered the regression equation; in DM1 group it was HOMA-IRI, PPG and DPG that entered the regression equation; in DM2 group it was FPG and HOMA-beta that entered the regression equation.@*CONCLUSION@#There exists a flow-mediated vasodilatation dysfunction in patients of newly diagnosed IGT and T2DM. Effect of relevant factors on FMD differs with different glucose levels.

Effects of rosiglitazone on the IMTc and serum MMP-9 levels in newly diagnosed type 2 diabetic patients / 中南大学学报(医学版)

OBJECTIVE@#To investigate the change of carotid intima-media thickness (IMTc) and serum matrix metalloproteinases-9 (MMP-9) levels in newly diagnosed Type 2 diabetic patients, and to analyze the relationship between MMP-9 and IMTc; at the same time, to assess the effect of rosiglitazone on IMTc and MMP-9 levels.@*METHODS@#Fifty-eight patients with Type 2 diabetes mellitus were selected in our study, and 25 healthy adults served as normal controls. Diabetic patients were divided into 2 groups: Group A (31 subjects) were treated with rosiglitazone (4 mg/d), and Group B (27 subjects) were treated with metformin alone (500 approximately 1,500 mg/d). They all received the treatment for 3 months. The IMTc was measured by high resolution ultrasonography, and the serum MMP-9 was determined by enzyme linked immunosorbent assay (ELISA) to assess the relationship between IMTc and MMP-9.@*RESULTS@#The mean level of serum IMTc and MMP-9 in Type 2 diabetic patients was significantly higher than that in healthy adults (P < 0.05). After treatment with rosiglitazone and metformin, IMTc and serum MMP-9 levels decreased significantly (P < 0.05). There was no obvious change in IMTc and serum MMP-9 levels in group B before and after the treatment (P = 0. 071, P = 0.065). Using multiple linear stepwise regression analysis, the significant correlation between IMTc and HbA1C, BMI, WHR, HDL-C, MMP-9 were discovered.@*CONCLUSION@#IMTc and MMP-9 levels increase in newly diagnosed Type 2 diabetic patients, suggesting that there is closely relationship between serume MMP-9 levels and early diabetic macrovascular disease. IMTc and MMP-9 can be reduced significantly in the newly diagnosed diabetic patients after being treated with rosiglitazone, which may be one of the protective mechanisms of vascular vessels.

Isolation, induction and differentiation of human blood-derived endothelia progenitor cells / 中南大学学报(医学版)

OBJECTIVE@#To determine the biological traits and optimal condition for the induction and differentiation of endothelial progenitor cells from peripheral blood in healthy adults.@*METHODS@#Mononuclear cells isolated from peripheral blood of healthy adults were cultured in M199 medium supplemented with VEGF, bFGF, IGF-1, and EGF. The appearing time of cell clusters or spindle-shaped cells was recorded respectively. Attached spindle-shaped cells were detached and labeled with a series of antibodies against blood vessel endothelial-specific markers.@*RESULTS@#Attached spindle-like cells appeared 4 days after the culture, cell clusters were observed at 5 to 8 days, and cord-like structure was formed by 10th day. These cells expressed endothelial-specific markers.@*CONCLUSION@#Endothelial progenitors cells were derived from mononuclear cells of peripheral blood, which can be induced into endothelial cells at specific conditions.

Effects of simvastatin on TGF-beta system of diabetic rat kidneys / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effects and mechanism of renal benefit of simvastatin on diabetic rat kidneys.@*METHODS@#Twenty STZ-induced SD rats and 10 normal rats were assigned to diabetic rat (DM) group, simvastatin [ 4 mg/( kg x d) ] treatment (S) group and normal control (C) group. Immunohistochemistry, RT-PCR and western-blot were employed to examine the changes of the mRNA and protein expression of TGF-beta1 and Tbeta II R in the kidneys of the rats.@*RESULTS@#Compared with the normal control group, both the mRNA and protein expression of TGF-beta1 and Tbeta II R in the diabetic rat group and treatment group were significantly increased (P < 0.05). Compared with the diabetic rat group, simvastatin could markedly decrease the mRNA and protein expression of TGF-beta1 and Tbeta II R (P < 0.05).@*CONCLUSION@#Simvastatim may play a protective role in the diabetic kidneys by down-regulating TGF-beta1 and Tbeta II R and inhibiting the TGF-beta signal pathway.