Objective To investigate the ultrastructure changes of pancreatic islets alpha cells in rats with impaired glucose tolerance.Methods 16 Wistar rats were randomly assigned into normal glucose tolerance group(NGT group,n =8)and impaired glucose tolerance group(IGT group,n =8).NGT group were fed with routine diet and IGT-group were fed with high-sugar high-fat diet.At the 12th week,IGT models were considered successful if the level of 2-hour peripheral blood glucose(2 h PG)was between 7.8 and 11.1 mmol/L in oral glucose tolerance test and continued for more than a week.The pancreas tail of animals in the NGT group and IGT group rats was removed for ultrastructure observation.Alpha cells were observed under transmission electron microscope.Results The big-plump secretory granule with aureole appeared in pancreatic islets alpha cells of NGT group,and it contained a round dense core.The nuclear chromatin and contents were evenly distributed.Mitochondria and en-doplasmic reticulum arranged tightly and orderly.Compared with NGT group,the quantities of secretory granule increased significantly in IGT group.The gap between dense core and membrane was narrower.The quantities of mitochondria and rough endoplasmic reticulum increased significantly.There was no significant change for the structure of the other cell organelle.Conclusion The pathogenesis of IGT is associated with ultrastructural changes and dysfunction of pancreas islets alpha cells.

Objective To investigate the level change of glucose excursion , 8-iso prostaglandin F 2α( 8-iso-PGF2α) in type 2 diabetes mellitus(T2DM), and its effects on nonalcoholic fatty liver disease (NAFLD).Methods A total of 62 inpatients with type 2 diabetes mellitus including T2DM group(DM, 30 cases) and T2DM with NAFLD group(DM+NAFLD, 32 cases) were recruited from October 2012 to November 2013 , normal glucose tolerance group had normal glucose tolerance ( NGT;30 cases ) with normal physical examination results.The age, gender, duration, blood pressure, body mass index (BMI), and blood lipid among three groups were no statistical difference .The clinical data of each patient were collected by professional people .Blood lipids and glycated hemoglobin (HbA1c) were detected.Oral glucose tolerance test (OGTT) were taken in all groups.The Postprandial glucose excursion (PPGE), blood glucose standard deviation (SDBG), mean blood glucose and (MBG) and HbA1c were used to evaluate the glucose excursion.Serum 8-iso-PGF2αwas detected by enzyme-linked immunosorbent (ELISA) to evaluate oxidative stress.Inter-group com-parison was conducted with analysis of variance ( ANOVA) .Correlation analysis was used to evaluate the factors of influence .Results⑴The levels of PPGE[(7.34 ±2.23) mmol/L vs (8.52 ±2.43) mmol/L],SDBG[(2.43 ±0.90) mmol/L vs (2.80 ±0.78) mmol/L], MBG[(11.06 ±1.76) mmol/L vs (13.65 ±2.83) mmol/L], and HbA1c [(7.59 ±1.02)% vs (8.05 ±1.52)%] in T2DM group and T2DM with NAFLD group were significantly higher than that in NGT group (both P <0.05); and that in NAFLD group have significantly rise than that in DM group ( P <0.05), but there was no significant difference in HbA 1c between groups.⑵ The level of serum 8-iso-PGF2αwas gradually increased from NGT group [(33.45 ±8.60) pg/ml], DM group [(47.33 ±15.30) pg/ml], to DM +NAFLD group [(56.07 ±13.10) pg/ml], with statistically significant difference ( P <0.05);⑶The Person corre-lation analysis showed that the content of serum 8-iso-PGF2αwas positively correlated with PPGE, SDBG, and MBG ( r =0.796, 0.778 , 0.712 , P <0.01 ) .Conclusions Serum 8-iso-PGF2αis a better parameter to reflect the status of body oxidative stress .The level of oxidative stress is increased with the increase of glucose excursion in T 2DM, which is the important mechanism of its complica-tions of NAFLD.

Objective To investigate the effect of glucagon-like peptide-1 agonist (exendin-4) on expression of glucose transporter 4 (GLUT4) in the skeletal muscle of rats with impaired glucose tolerance (IGT).Methodis 54 Wistar rats were randomly assigned into normal glucose tolerance group (NGT group,n =18) and impaired glucose tolerance group(IGT group,n =36).The rats in NGT group were fed with routine diet and the rats in IGT group were fed with high-sugar high-fat diet.At the 12th week,IGT models were tested successful.Then,half of the rats were allocated to intervention group (Ex group) and the rest were set as IGT control group.The rats in Ex group were subject to exendin-4 subcutaneous injection (5 μg/kg,twice daily).Each rat in NGT group and IGT control group was given the same volume of saline as injection.FBG and 2 h BG were measured before intervention and after 4 weeks.The expression of GLUT4 mRNA and GLUT4 in the skeletal muscle were respectively measured by real time quantitative polymerase chain reaction and immunohistochemistry.Inter-group comparison was conducted using analysis of variance (ANOVA) and least square deviation-test (LSD-t).Results Before intervention,compared with NGT-group,the 2 h BG in IGT control group and Ex group were higher,the GLUT4 mRNA of skeletal muscle in IGT control group and Ex group were lower (respectively P ＜ 0.05).The skeletal muscle cells in IGT control group and Ex group were less colored while the skeletal muscle cells in NGT group were colored extensively,and more colored granules.After 4 weeks of exendin-4 intervention,compared with IGT control group and Ex group of non-intervention,the 2 h BG level in Ex group was lower and the expression level of GLUT4 mRNA of skeletal muscle was higher (respectively P ＜ 0.05).After intervened with exendin-4 for 4 weeks,the GLU protein mainly expressed in cytoplasma of skeletal muscle cells.Its expression was higher in Ex group than in IGT group and in Ex group before intervention.Conclusion Exendin-4 may up-regulate the expression of GLUT4,increase glucose intake of the skeletal muscle,and reduce postprandial blood sugar.

Objective To observe and compare the changes of weight,blood pressure,glycemia, blood lipid,serum uric acid,insulin sensitivity and insulin function of laparoscopic sleeve gastrectomy (LSG)for the treatment of type 2 diabetes mellitus(T2DM)with obesity,and analysed mechanism of meta-bolic surgery.Methods Changes of weight,blood pressure,glycemia,blood lipid ,serum uric acid,in-sulin sensitivity and insulin function of 8 type 2 diabetes mellitus(T2DM)with obesity patients undergoing LSG were retrospectively analyzed.Weight,BMI,EWL%,average systolic and diastolic pressure were measured 1,2,3,6 months and fasting plasma glucose (FPG),glycosylated hemoglobin (HbAl),insulin sensitivity parameter:insulin resistance index(HOMA-IR),insulin sensitivity index(ISI),insulin area un-der the curve (AUCI),insulin function parameter:glucose disposition index(DI)were measured 1,6 months after operation and compared with preoperative levels.Meanwhile medication change was observed af-ter operation and compared with preoperation.Results ⑴Weight ,BMI at 1,2,3,and 6 months after surgery showed a decreasing trend and EWL% showed a decreasing trend over time .⑵ FBG、HbA1c showed a decreasing trend after operation.Blood lipid,uric acid of 3 patients with dyslipidemia and hyperu-ricemia decreased to normal 1 month after operation.⑶ HOMA-IR,AUCI except for 1 patient showed a de-creasing trend and ISI was contrary after operation.DI ,trends were different.⑷7 patients stopped all an-tidiabetic drugs 6 months after operation,1 patient injected insulin with good glycemic control.Blood pres-sure of 5 patients with hypertension was normal without antihypertensive drugs 6 months after operation. Conclusions LSG procedure can significantly increase insulin sensitivity of T2DM with obesity,thereby improve glycemia,cholesterol,uric acid and further promot weight loss.

ObjectiveTo study the association of SNP276 in adiponectin gene with impaired glucose regulation (IGR) complicating with the different component numbers of metabolism syndrome (MS) in population of IGR in Han people of Shanxi region.MethodsThe study population consisted of 256 subjects with IGR which was composed of 123 subjects with component numbers of MS ＜2 (IGRA) and 133subjects with component numbers of MS≥2 (IGRB),and 128 subjects with normal healthy (Normal) who were Chinese Hans residents and in Shanxi province.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was applied to test the adiponectin SNP276G/T polymorphism.ResultsThe distributions of genotypes and alleles of SNP276 both displayed significant difference among the three groups ( x2 =16.893,P =0.002 ; x2 =18.149,P =0.000).In the IGRA,the SNP276 non-TT ( GT + GG) genotype was no difference which increased risk of complicating with the different component numbers of MS( P =0.781,P =0.809).In IGRB,the SNP276 non-TT ( GT + GG) genotype was associated with increased risk of complicating with the different component numbers of MS,and after age and sex-adjusted,there was significant difference ( P =0.007,P =0.007).ConclusionsSNP276 in APM1 increased the risk of complicating with the components of MS in population with impaired glucose regulation in Han people of Shanxi region.

Objective To assess the relationship between serum vitamin D and autoimmune thyroid disease.Methods Subjects included total 520 persons receiving regular health examination,and serum calcium,phosphorus,parathyroid hormone (PTH),thyroid peroxidase autoantibody (TPOAb) and 25-dihydroxy vitamin Ds was measured.The incidence of 25-dihydroxy vitamin D3 deficiency (≤30 μ g/L)was observed.The relationship between 25-dihydroxy vitamin D3 deficiency and autoimmune thyroid disease was analyzed.Results The serum 25-dihydroxy vitamin D3 of all the subjects was (24.47 ± 7.21) μ g/L,and the incidence of 25-dihydroxy vitamin D3 deficiency (≤30 μg/L) was 61.15％ (318/520),and the positive rate of TPOAb was 21.54％ (112/520).The proportion of TPOAb ＞ 50 kU/L or ＞ 100 kU/L in subjects with 25-dihydroxy vitamin D3≤30 μ g/L was higher than that in subjects with 25-dihydroxy vitamin D3 ＞ 30 μg/L [25.79％(82/318) vs.19.80％(40/202) and 9.43％(30/318) vs.4.46％(9/202)],and there was significant difference (P ＜ 0.05).The relationship between 25-dihydroxy vitamin D3 and TPOAb was assessed and showed significant inverse correlation (r =-0.13,P ＜0.05).Conclusions Vitamin D deficiency is very common in the population,and autoimmune thyroid disease is related with vitamin D deficiency,which may has impact on the body's immune regulation.Specific mechanism and whether vitamin D supplementation can intervene and treat autoimmune thyroid disease need further study.

Objective To assess the role of the different pancreatic islet β cell function index in the evaluation of glucose metabolism in different duration of type 2 diabetes mellitus (T2DM).Methods Normal glucose tolerance subjects without diabetes family history (NC group,48 cases) and T2DM patients (182 cases) were enrolled.The T2DM patients were divided into three groups:less than 5 years group (DM ＜5 group,74 cases),5-10 years group (DM5-10 group,51 cases) and more than 10 years group ( DM ＞10 group,57 cases).Oral glucose tolerance test (OGTT) and insulin release test were taken in all groups.Insulin resistance index (HOMA-IR) and whole body insulin sensitivity index [ISI(Matsuda)] were used to estimate insulin sensitivity,and early insulin secretion index ( △ I30/ △ G30) and glucose disposition index (DI) were used to evaluate the function of pancreatic islet β cell.Results HOMA-IR was increased and ISI (Matsuda) was decreased in DM ＜5 group,DM5-10 group and DM ＞10 group compared with those in NC group [HOMA-IR:8.78 ± 7.12,8.08 ± 3.67,7.84 ± 5.08 vs.4.76 ± 3.43;ISI(Matsuda):46.78 ± 29.00,36.71 ± 16.67,38.86 ±21.72 vs.61.13 ± 32.08,P ＜ 0.05],however,there was no significant difference among DM ＜5 group,DM5-10 group and DM ＞10 group.△ I30/ △ G30 and DI were decreased in DM ＜5 group,DM5-10 group and DM ＞10 group compared with those in NC group [ △ I30 △ G30:( 68.41 ± 361.52 ),(4.31 ± 3.42 ),(7.70 ± 5.78 ) mU/mmol vs.(92.65 ± 309.29) mU/mmol;DI:0.0421 ± 0.0123,0.0412 ± 0.0123,0.0363 ± 0.0116 vs.0.1151 ± 0.0236,P ＜ 0.05 ],and there was no significant difference in △ I30 / △ G30 among DM ＜5 group,DM5-10 group and DM ＞10 group,however,DI was decreased in DM＞10 group compared with that in DM＜5 group and DM5-10 group (P＜0.05).ConclusionsHOMA-IR,ISI (Matsuda),△I30/△G30 are not sensitive to evaluate the insulin resistance of different duration.DI can reflect the glucose utilization of pancreatic islet β cell earlier and the ability to regulate blood sugar steady state changes.

Objective To observe the effect of oxidized low-density lipoproteins (Ox-LDL),15-Deoxy-△ 12,14-prostaglandin J2 ( 15d-PGJ2 ),leukotrienes B4 ( LTB4 ) on mRNA expressions of peroxisome proliferator activated receptor γ2 ( PPARγ2 ),receptor activator of NF-κB ligand (RANKL),alkaline phosphatase ( ALP),and osteoprotegerin(OPG) in osteoblastic cells of rats; and to investigate the influence of these PPARγ2 endogenous ligands on bone metabolism.Methods Rat osteoblastic cells were cultured in vitro for 24 h in medium with different PPARγ2 endogenous ligands at various concentrations ( the final concentrations of Ox-LDL were 0,12.5,25,50μg/ml; the final concentrations of 15 d-PGJ2 were 0,10,20,30 μmol/L; the final concentrations of LTB4 were 0,0.1,1.0,10 μ mol/L).RT-PCR was performed to determine the mRNA expressions of PPARγ2,RANKL,ALP,and OPG in osteoblastic cells.Results RT-PCR analysis showed that Ox-LDL,15d-PGJ2,and LTB4 all down-regulated the mRNA expressions of RANKL,ALP,and OPG,while up-regulated the mRNA expressions of PPARγ2 in osteoblastic cells in a dose-dependent manner.Significant differences were found in interclass comparisons( P＜0.05 or P＜ 0.01 ).Conclusions These findings suggest that Ox-LDL,15d-PGJ2,and LTB4 suppress the expressions of osteogenic genes through activating the transcription activity of PPARγ2,and this may be a plausible mechanism of senile osteoporosis.

Objective To study the effect of pioglitazone on the differentiation and function of rat osteoclast-like cells (OLC), and to probe the relationship between activated PPARγ2 and osteoclasts. Methods On day 1 of OLC formation from nonadherent bone marrow ceils (BMC) obtained from rats induced by M-CSF and receptor activator of NF-кB ligand (RANKL), 1, 5 and 10μmol/L pioglitazone hydrochloride was added. RT- PCR was performed to determine the mRNA expressions of PPARγ2 and receptor activator of NF-кB (RANK) on day 3, 5 and 7 during incubation, the number of tartrate-resistant acid phosphatase (TRAP)-positive cells,the number of bone resorption pits and the ratio of its area on dentin slice were counted, the activity of TRAP and the mean fluorescence intensity of integrin β3 (CD61) of OLC were also measured. Results (1) The effect on the differentiation of OLC: The addition of pioglitazone at the start of the culture period induced a dose-dependent decrease in TRAP-positive OLC and the activity of TRAP (P < 0.01 or P < 0.05) ; the mRNA expression of PPARγ2 was up-regulated by 5 and 10 μmol/L pioglitazone in the early stage of incubation and attenuated with thematuration of OLC on the contrary, however, the expression of RANK was down-regulated by 5 and 10 μmol/L piolitazone in every stage of incubation (P < 0.05 or P < 0.01), combined with decrease in TRAP-positive OLC from day 3 by 10 μmol/L pioglitazone. (2) The effect on the function of OLC: the number of bone resorption pits and the ratio of its area on dentin slice were decreased in groups of 5 and 10 μmol/L pioglitazone (P < 0.01 orP < 0.05), no obvious change was noted in the group with 1 μmol/L pioglitazone compared with the control group; the mean fluorescence intensity of CD61 were down-regulated in groups of 5 and 10 μmol/L pioglitazone (P < 0.05 or P <0.01). Conclusion Activation of PPARγ2 pathway by pioglitazone could partially inhibit differentiation and function of OLC derived from rat BMC.

OBJECTIVETo compare and explore metabolic risk factors related to type 2 diabetes mellitus (T2DM) and the development of nonalcoholic fatty liver disease (NAFLD).

METHODSA total of 389 in-patients with T2DM (DM group, 204 cases) and T2DM with NAFLD (DM+NAFLD group, 185 cases) were enrolled in the study between October 2012 and July 2013. Clinical data collected for analysis included levels of blood lipids, liver function markers, uric acid (UA) and insulin, as well as results from the oral glucose tolerance test (OGTT) and C peptide releasing test. Improvements in insulin level, C peptide secretion index [HOMR-IR(CP)] and whole body insulin sensitivity index (ISIcomp) were used to estimate insulin sensitivity. Improvements in insulin level, C peptide secretion index [HOMR-islet(CP)], early insulin secretion index (delta I₃₀/delta G₃₀), correction of the islet beta cell function index (MBC1) and glucose disposition index (DI) were used to evaluate the function of pancreatic islet 1 beta cells. The t-test and repeated measures ANOVA were used for statistical analyses. Risk factors of T2DM with NAFLD were assessed by using logistic regression analysis.

RESULTSCompared with the DM group, the DM+NAFLD group had higher body mass index (BMI) and levels of triglycerides, alanine aminotransferase (ALT), aspartate aminotransferase, gamma glutamine transferase and UA (all P < 0.05), but lower age and level of high density lipoprotein cholesterol (both P < 0.05). Moreover, the DM+NAFLD group had higher postprandial blood glucose levels at 30 min (10.88 ± 2.87 mmol/L vs. 12.18 ± 2.79 mmol/L, t =-3.32), 60 min (14.65 ± 3.69 mmol/L vs. 15.99 ± 3.12 mmol/L, t =-3.46), 120 min (16.56 ± 5.11 mmol/L vs. 17.65 ± 4.29 mmol/L, t =-2.81) and 180 min (13.92 ± 5.10 mmol/L vs. 14.71 ± 4.91 mmol/L, t=-2.02) (all P < 0.05). The DM+NAFLD group had higher postprandial insulin levels at 60 min (28.62 ± 23.51 muIU/ml vs. 36.91 ± 33.47 aIU/ml, t =-3.46) and 120 min (36.36 ± 25.60 muIU/mL vs. 44.38 ± 34.95 muIU/mL, t =-3.35) (both P < 0.05). The DM+NAFLD group had higher postprandial C peptide levels at 30 min (2.74 ± 1.70 ng/mL vs. 4.30 ± 6.51 ng/ml, t =-4.97), 60 min (4.17+/-2.49 ng/ml vs. 5.19 ± 2.96 ng/ml, t =-3.29) and 120 min (6.08 ± 2.79 ng/ml vs. 6.76 ± 3.10 ng/ml, t =-2.19) (all P < 0.05). The DM+NAFLD group had higher HOMA-IR(CP) (1.505 ± 0.004 vs. 1.507 ± 0.005, t =-2.208, P less than 0.05), but lower ISIcomp (90.09+/-69.31 vs. 59.93 ± 24.52, t =5.608), MBCI (4.68 ± 4.31 vs. 3.83 ± 2.41, t =2.365) and DI (35.40 ± 71.83 vs. 15.37 ± 13.93, t =3.730) (all P < 0.05). Logistic analysis showed that BMI, ALT, postprandial level of C-peptide at 30 min, and UA were the major risk factors of T2DM with NAFLD (OR =1.208, 2.080, 1.041, and 1.005, respectively; all P < 0.05).

CONCLUSIONPatients with a propensity for developing nonalcoholic fatty liver disease may experience earlier open of type 2 diabetes. T2DM patients with NAFLD have more severe glucose metabolism disorders.

Adult ; Aged ; Aged, 80 and over ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; complications ; metabolism ; Female ; Humans ; Insulin ; blood ; Lipids ; blood ; Male ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; complications ; metabolism ; Risk Factors