Objective: To investigate the apoptosis of renal cells induced by tumor necrosis factor alpha (TNF-α) and nuclear factor-κB (NF-κB) in diabetic rats and intervention of rapamycin. Methods: A total of 20 rats (Goto-Kakizaki rats) with type 2 diabetes mellitus (T2DM) were randomly and equally divided into DM model group (DM group) and rapamycin treatment group (DMR group, received rapamycin treatment after DM model was established); another 10 Wistar male rats were regard as normal control group. Apoptosis of renal cells, expression levels of TNF-α and NF-κB and levels of blood lipids, blood glucose were measured in all groups after four weeks and eight weeks. Results: Four and eight weeks After model was established, compared with normal control group and DMR group, there were significant increase in renal cells apoptosis [RCA, four weeks: (0.217±0.031), (0.272±0.031) vs. (0.545±0.031), eight weeks: (0.358±0.031), (0.350±0.031) vs. (0.811±0.031)] and expressions of NF-κBp65 [OD: four weeks: (0.160±0.027), (0.131±0.027) vs. (0.411±0.027), eight weeks: (0.232±0.027), (0.275±0.027) vs. ( 0.634±0.027)] and TNF-α [OD: four weeks: (0.242±0.027), (0.275±0.027) vs. (0.617±0.027), eight weeks: (0.385±0.027), (0.342±0.027) vs. (0.912±0.027)] in DM group (P<0.01 all). Correlation analysis indicated that there were positive correlations between renal NF-κBp65 and TNF-α, among RCA and TNF-α, NF-κBp65 (r=0.956, 0.953, 0.886，P<0.01 all).

BACKGROUND: The islet cell transplantation has provided a solid basis for diabetic therapy, but the insufficient donor limits its development.OBJECTIVE: improving the method of isolating and purifying islets to observe the transplantation effect.DESIGN: A laboratory animal research.SETTING: Key Laboratory of Animal and Department of Cell Biology, China Medical University.MATERIALS: The experiment was carried out in the Key Laboratory of Animal and Department of Cell Biology in China Medical University between January and October in 2006. Donors were Wistar rats of either gender, weight 250-300 g;Acceptors were SD male rats, weight 180-220g. The two kinds of rats were all common closed population and from the Experimental Animal Department of China Medical University (The Admission Number of Experimental Animal Institute is SYXK(LIAO)2003-0013).METHODS: ①Isolation and exaltation of islet cells as well as the functional evaluation of pancreas: After etherisation, the Wistar rat without fasting was executed. A little cut was made on the beginning of the biliary pore, then the little cut lumbar anesthesia ductus, which were connected with a 1-mm-diameter syringe and full of cold collagenase solution (1.5 g/L), was inserted directly to dilate pancreas thoroughly. The pancreatic gland was isolated and digested in the water of centrifuge, when doing that, 1 mol/L NaOH was put interruptedly into the centrifuge tube to keep the pH value of the solution at 7.8±1.0. The rat pancreas purified by centrifugation of Ficoll density gradient: The identification of purified islets was evaluated by dithizone staining. The viability of islet was assessed by fluorescence staining of aridine orange and propidium iodide. The motility rate=the total number of live cells/(the total number of live cells + the total number of dead cells)×100%. Pancreatic activity was calculated: insulin release index=the level of insulin at the third hour (high concentration glucose)/the level of insulin at the second hour (low concentration glucose). ②The blood from vena caudalis of SD rat was sampled and measured the blood sugar after the intraperitoneal injection of streptozocin. The rat was diagnosed as DM when blood sugar was more than 16.7 mmol/L twice without fasting. The DM rats were divided into two groups, every group 8 rats. The experimental group rats were injected about 1 000 islet cells into the location below renal capsule, and the control group rats were injected the same volume of 1640 cultu re solution. Eight normal rats, whose glucose concentration ≤ 5.5 mmol/L, were taken randomly as normal controlled group. The blood sugar was measured every day after the surgery. The blood sugar less than 11.1 mmol/L without fasting was taken as the sign of successful islet transplantation. Intravenous sugar tolerance test was applied to the rats of normal control group, DM control group and experimental group 3 days after islet transplantation. Fasting for 12 hours before test, the blood sugar was measured at 0, 15, 30, 60, 90 and 120 minutes.MAIN OUTCOME MEASURES: Purity quotient, survival rate and activity of islet cells.RESULTS: All 24 SD acceptor rats were involved in the result analysis without miss.①The total number of purified islets of one pancreas was (1 150±141) in well morphology. The purity of islets was more than 95%. The viability of islets was more than 98%. ②The insulin secretion response to glucose challenge in vitro showed the mean value of insulin in the low-glucose medium was (70.5±6.9) mlU/L, while that of high-glucose medium was (321.4±11.6) mlU/L, the insulin release index was 4.6±0.52, that meant the beta cell of islet functioned well. ③The blood glucose level and the insulin level in plasma of the transplanted recipients restored to normal 3 days after transplantation. The survival period of transplanted islets was (6±2) days. But there was not any change in the concentration of blood sugar in the control group (16.7 mmol/L). The intravenous glucose tolerance test showed the identical outcome between the islet splantation group and the normal control group.CONCLUSION: There are high yield and high purify of islet cells in rats, which are isolated by in situ perfusion and purified by Ficoll density gradient centrifugation.

Objective: To study effect of small ubiquitin related modifier protein 1 (SUMO1) in inflammatory reactions mediated by tumor necrosis factor (TNF)-α and nuclear factor (NF)-κB in myocardial damage of rats with type 2 diabetes mellitus (T2DM). Methods: A total of 20 Goto-Kakizaki (GK) rats with spontaneous diabetes mellitus (DM) were randomly divided into group DM1 (pure DM group, n=10) and group DM2 (DM+high-fat diet group, n=10), and another 10 normal Wistar rats were regard as healthy control group. Expressions of SUMO1, TNF-α and NF-κB were measured by immunohistochemical method. Results: 1. Levels of blood glucose and TG in group DM1 and group DM2 were significantly higher than those of healthy control group, and those of DM2 group were higher than of DM1 group ,P<0.05 all; 2. Myocardial cells lined up in order and there was no hypertrophy in group DM1; but those in group DM2 showed cells loosely lined up and hypertrophy under light microscope; 3 Immunohistochemical assay indicated that expression of SUMO1 in group DM2 and DM1 group were significantly higher than those of healthy control group [(44.5±1.1) vs. (27.2±2.2) vs. (21.7±3.0)], and of group DM2 was significantly higher than that of DM1 group (P<0.01 all); expression of TNF-α in group DM2 and group DM1 were significantly higher than that of healthy control group [(27.5±1.5) vs. (20.2±2.7) vs. (13.1±1.6)], and of DM2 group was significantly higher than that of group DM1 (P<0.01 all)；expression of NF-κB in group DM2 and group DM1 were significantly higher than that of healthy control group [（30.1±1.7）vs.40.7±1.5）vs.（16.0±2.6）], but of group DM1 was significantly higher than that of group DM2 (P<0.01 all). Conclusion: There are obvious metabolic disorders of glucose and lipid in T2DM rats, and complicated morphological changes of myocardial tissues similar to myocardial lesions in DM humans; the expressions of SUMO1, NF-κB and TNF-α significantly increase, suggest SUMO1 takes part in inflammatory reaction mediated by NF-κB, TNF-α in myocardial lesion of rat with T2DM，and may inhibit NF-κB, possesses effect of protect myocardium.

Objective: To study relative risk factors for diabetic nephropathy (DN). Methods: A total of 238 patients diagnosed as type 2 diabetes mellitus (DM) were enrolled in the study. According to urine microalbuminuria to urine creatinine ratio (UACR), patients were divided into pure DM group (group DM1, n=90), early diabetic nephropathy group (group DM2 , n=73) and clinical diabetic nephropathy group (group DM3 ,n=75). Clinic data of all patients were collected; Fasting blood glucose (FBG), 2h postprandial blood glucose (2hPB), blood lipids, uric acid (UA), fibrinogen (Fg) and glycosylated haemoglobin (HbA1c) were measured in all patients, and their correlations with DN were analyzed. Results: Compared with group DM1, the course of disease in DM [(7.25±6.29) years vs. (10.25±7.67) years vs. (13.53±7.82) years], levels of FBG [(8.46±2.52) mmol/L vs. (9.52±3.38) mmol/L vs. (10.82±3.30) mmol/L], 2hPB [(18.40±5.64) mmol/L vs. (20.27±5.94) mmol/L vs. (22.59±6.14) mmol/L], HbA1c [(7.96±1.65) % vs. (8.60±1.76) % vs. (9.55±2.09) %], triglyceride [TG, (1.72±0.86) mmol/L vs. (2.34±1.87) mmol/L vs. (3.16±1.85) mmol/L], Fg [(3.49±0.93) g/L vs. (3.88±1.21) g/L vs. (4.99±2.10) g/L] and UA [(295.42±52.34) μmol/L vs. (324.18±96.29) μmol/L vs. (351.23±56.88) μmol/L] significantly increased in group DM2 and group DM3 in order (P<0.01~0.001). Logistic gradual regression analysis indicated that course of DM, HbA1c, TG, Fg and UA were risk factors for DN (OR=1.008~1.910, P<0.01~0.001). Conclusion: The course of DM, blood glucose, blood lipid, uric acid and fibrinogen are risk factors of diabetic nephropathy; increased UACR reflects progress of patient’ condition in DM patients, its detection is used for diabetic prognosis and treatment.

Objective: To study influence of overweight and obesity on blood coagulation and metabolic disorders in patients with type 2 diabetes mellitus (T2DM). Methods: A total of 248 preliminary diagnosed T2DM patients were selected. According to body mass index (BMI), they were divided into normal BMI control group (n=95), overweight group (n=87) and obesity group (n=66). Blood lipids, blood glucose and fasting insulin (FINS) were measured in all patients and homeostasis model-insulin resistance index (HOMA-IR) was calculated then. Statistical analysis was performed. Results: Compared with normal BMI control group, there were significant increase in fibrinogen [(3.37±0.55) g/L vs. (4.04±0.70) g/L vs. (5.20±0.69) g/L], urine microalbumin [(14.46±8.90) mg/g vs. (47.33±42.54) mg/g vs. (104.45±60.78) mg/g], fasting blood glucose [ (7.15±0.97) mmol/L vs. (8.84±1.81) mmol/L vs. (10.06±2.28) mmol/L], FINS [(10.09±8.21) IU/ml vs. (14.33±15.55) IU/ml vs. (19.69±10.86) IU/ml], HOMA-IR[(3.19±2.59) vs. (5.51±5.38) vs. (8.48±4.62)], TG, TC and LDL-C levels in overweight group and obesity group, and the more BMI patients were, the higher these indicators were; There were significant decrease in plasma prothrombin time [(13.33±0.69)s vs. (12.74±0.69)s vs. (11.43±0.53)s], activated partial thromboplastin time [ (37.32±2.31)s vs. (36.55±2.41)s vs. (34.61±1.53)s] and HDL-C [(1.54±1.12) mmol/L vs. (1.27±0.41) mmol/L vs. (1.09±0.28) mmol/L] in overweight group and obesity group（P<0.05 all）. Conclusions: Overweight and obesity aggravate coagulation and metabolic disorders in patients with type 2 diabetes mellitus. It also aggravates degree of insulin resistance, the more BMI patients are, the more serious they are.

Objective To evaluate the effect of positive experience intervention on anxiety, depression and quality of life among rural empty nesters. Methods A total of 115 rural empty nesters were divided into the intervention group (n=58) and the control group (n=57). Both groups received routine community activities. In addition, participants in the intervention group received positive experience intervention. Geriatric Depression Scale, Self-Rating Anxiety Scale and 12-item Short-Form Health Survey were adopted to measure the related items before and after the intervention. Results The intervention group showed a significant decrease in anxiety (1.54±0.07) and depression (7.28±3.82), compared with the control group (1.64±0.09, 11.81±6.38), the difference was significant (t=-5.960,-4.613, P<0.01). The Physical Component Summary (PCS) and Mental Component Summary (MCS) was 47.01±6.47 and 52.13±6.22 in the intervention group, which were significantly higher than those in the control group (41.37±8.71, 48.71±6.13), the differences were significant (t=3.937, 2.970, P < 0.01). Conclusions Positive experience intervention can decrease anxiety and depression, and improve quality of life of rural empty nesters.

Objective To characterize the changes of retinal structure and visual field mean defects (MD) in early Parkinson's disease (PD) and the related factors.Methods Twenty-five patients with early stage PD and 33 normal controls were enrolled in this study.PD patients were evaluated by unified Parkinson's disease rating scale (UPDRS) and Hoehn-Yahr (H-Y) stage in off period.All subjects undertook retina of retinal nerve fiber layer (RNFL) and macular examinations by optical coherence tomography (OCT),and MD by automatic visual field analyzer.Results (1) The average thicknesses of RNFL and C11 quadrant RNFL were thinner in PD group [(96.2 ±7.6) μm and (124.4 ± 18.4) μm] than in healthy controls [(102.6 ± 5.0) μm and (135.4 ± 21.8) μm,respectively,P =0.000 and P =0.047].Moreover,PD patients had a thinner average thickness of macular[(277.2 ±9.6) μm vs (285.8 ± 12.6)μm,P=0.006],and smaller macular volume compared with controls [(10.0 ± 0.3) mm3 vs (10.3 ±0.4) mm3,P =0.006].However,there was no significant difference in MD between PD and control subjects [(0.43 ± 2.75) dB vs (-0.18 ± 1.41) dB,P =0.322].(2) Pearson's correlation analysis showed that H-Y stage was negative correlated with the average RNFL thickness (r =-0.569),average macular thickness and volume (r=-0.501 and r=-0.417) in PD patients (all P＜0.05).Conclusions PD patients have thinner average thicknesses of RNFL,C11 quadrant RNFL and macular,smaller macular volume than those in normal controls.There were no significant differences in MD between two groups.There were negative relationships between H-Y stage,and retinal thickness and volume of PD patients.

Objective: To study expressions of small ubiquitin-related modifier protein（SUMO）4 (SUMO4), nuclear factor (NF)- κB and inhibitory factor of NF-κB (IκB) in kidneys of rats with type 2 diabetes mellitus (T2DM). Methods: A total of ten 40-week-old male Goto-Kakizaki (GK) rats （with spontaneous diabetes mellitus）of specific-pathogen free (SPF) grade, and ten 40-week-old male Wistar rats of SPF grade were selected. The lesion of renal tissue was observed by hematoxylin eosin (HE) staining. Expressions of SUMO4, NF-κB and IκB in renal tissue were observed by immunohistochemistry methods. Results: In the GK rats, glomerular capillary ball hypertrophy, basilar membrane slightly thickening; glomerular mesangial cells hyperplasia, hypertrophy and renal tubular epithelial cells hypertrophy were observed. Compared with normal Wistar rats, expression levels of NF-κB [(0.232±0.034) vs. (0.634±0.058)], IκB [(0.242±0.027) vs. (0.712±0.078)] and SUMO4 [(0.160±0.031) vs. (0.545±0.045)] significantly increased in renal tissue of GK rats (P<0.01 all). Conclusion: Compared with Wistar rats, expressions of NF-κB, IκB and SUMO4 significantly increase in renal tissue of GK rats, suggesting that SUMO inhibiting transcriptional activity of NF-κB may exist in kidneys of T2DM rats. Therefore, sumoylation may be a new therapeutic target for inhibit renal microvascular lesion of diabetic disease.

Objective To observe the therapeutic effect of glycosaminoglycans(GAGs)on diabetes mellitus with early nephropathy in order to find a new therapeutic approach to diabetic nephropathy. Methods 60 cases of type 2 diabetic nephropathy(albuminuria:30 to 300mg/24h,male/female:38/22,ages:43-70 years old, course of disease:1-30 years)without hypertension were selected. Some indexes were analyzed before and after administration of regular therapy in routine group or glycosaminoglycans group. The elderly group and non elderly group of diabetes nephropathy were compared. When the metabolism is stable, the levels of endothelin (ET), Netricoxide (NO)and urinary albumin excretion rate(UAER)were measured. Results After three months treatment, the levels of UAER were decreased significantly in both GAGs group and routine group(P＜0.01).After three months, UAER was decreased step by step, and there was no difference between the two groups. The levels of UAER had no change in regular group and there was significant difference between this group and the other two groups. In GAGs group, the levels of whole blood viscosity of medium shear rate 1,whole blood viscosity of medium sheer rate 2,whole blood viscosity of low shear rate were declined and serum NO increased significantly; that of plasma viscosity, whole blood reduced viscosity, ET were all decreased to some degree. Conclusion GAGs has the therapeutic effect on type 2 diabetic nephropathy patients with microalbuminuria because of decreasing UAER and reversing the development of DN. The benefit was positively correlated with the time of taking glycosaminoglycans.

Objective: To study risk factors for non-alcoholic fatty liver disease (NAFLD) and vascular erectile dysfunction (ED) in patients with metabolic syndrome (MS). Methods: A total of 18 096 subjects were selected from people undergoing physical examination from 2008 to 2009 in northern cities of China by random cluster sampling method, and analyzed the risk factors for NAFLD and ED. Results: The 18 096 cases with age 18~76 (46.8±10.1) years old，containing 10 096 (55.79%) males and 8 000 (44.21%) females. Awareness rate of MS was 8.33% and prevalence rate of MS in healthy adults was 21.18%. Most common components of MS were hyperuricemia (27%, 4838/18096), obesity and overweight (21%), hypertension (20%,) and dyslipidemia (17%) in turn. Body mass index (BMI, kg/m2) and waist/hip ratio (WHR) of all MS subgroups from high to low were ED group [(28.9±1.1), (1.26±0.03)], overweight or obesity group [(27.5±2.3), (1.31±0.03)], prediabetes group [(26.8±2.6), (1.03±0.03)] and hypertension group [(26.1±1.3), (0.90±0.04)] in turn. A total of 3 721 MS patients （20.56%）complicated with NAFLD; By means of NAFLD complicated by MS as dependent variable, Logistic regression analysis indicated that increased ALT, waist circumference（WC）, age, DM family history, LDL-C and BMI (β=1.004~0.479, P=0.000~0.016 in turn) were risk factors for NAFLD, and physical exercise and occupational physical work were protective factors for NAFLD. There were 106 ED males and its prevalence rate was 1.04%; Logistic regression analysis indicated that age, WC, LDL-C, DM family history and BMI (β=0.681~0.238, P=0.000~0.018 in turn) were risk factors for ED, and educational degree, physical exercise and occupational physical work were protective factors for ED. Conclusion: Risk factors for NAFLD and ED in MS were closely correlated with MS. It’s a new path to prevent and treat NAFLD and ED through correcting risk factors of MS.