Objective To investigate the effect of bcl-2 gene tr ansfection on the apoptosis and bioactivity of cultured islet cells. Met hods Adenovirus-mediated gene transfection was employed to transfect b cl-2 into the islet cells, the expression of bcl-2 was determined by RT-PCR a nd immunocytochemical staining, and the apoptosis of islet cells was detected by TUNEL, the viability was determined by trypan blue staining and islets function was evaluated by insulin detection. Results Bcl-2 protein exp ressed in 70% of transfected rat islet cells. The apoptosis rate of transfected cells reduced significantly compared with untransfected cells (6% vs. 22%). The viability of transfected cells was higher than that of untransfected cells (91% vs. 68%). Insulin secrete capacity of transfected islet cells in high glucose me dium was superior to untransfected cells. Conclusions Islet cel ls may be successfully transfected with adenovirus-mediated gene transfection t echniques. The apoptosis may be declined by bcl-2 gene transfection, so the isl et cells' function improved.

Objective To investigate the effect of transplantation of islet allograft with Bcl-2 gene transfection on diabetic rats. Methods Among 26 Wistar rats, 22 were built as model of diabetes, and were divided in to 3 groups. Islet cells expressing Bcl-2 by adenovirus-mediated Bcl-2 gene transfection were transplanted through the portal vein of streptozotocin-induced diabetic rats. The function and rejection of the transplanted islet allograft were evaluated by analysis of blood glucose level and histological examination after transplantation. Results 48h after transplantation, the level of blood glucose was significantly decreased in both transfected and untransfected islets transplantation groups. The euglycemia period of transfected islet cells transplantation group was longer than that of the untransfected islet cells transplantation group (16.4?4.3d vs 6.8?2.2d, P

Objective To construct recombinant human Bcl-2 adenoviral vector. Methods Sense human Bcl-2 cDNA fragment from plasimid pcDNA3 1/bcl-2 was cloned into adenovirus shuttle vector pAdCMV. pAdCMV/bcl-2 and plasmid pJM17 were cotransfected into 293 cells with lipofectamine 2000. Replication defective adenovirus was reproduced and purified. Adenoviral virus DNA was detected with polymerase chain reaction(PCR), and plaque assay was performed to determine the titer of virus. Results Recombinant adenovirus containing bcl-2 target gene was obtained. The titer of virus was 2 0?10 10 pfu/ml. Conclusion High titer of adenovirus containing target gene was successfully obtained by recombinant adenovirus vector.

Objective To investigate the impact of experimental stress on serum glucose , insulin, glucagon, cortisol, and ep-inephrine in diabetic rats and its intervention with insulin and fluoxetine .Methods Type 2 diabetic rats model was induced by feeding with high sugar and high fat diet , and intraperitoneal injection of streptozotocin (STZ).The rats were divided into normal group , dia-betic control group , diabetic treated with insulin , and diabetic treated with fluoxetine .All the rats were exposed to multiple stressors (forced swimming, cold stimulation, rotation, restrain, and crowding) for 8 weeks.The rat blood sera collected from each group were analyzed with multiple parameters including glucose , insulin, glucagon, cortisol, and epinephrine at the 4th and 8th week.Results After experimental stress, the levels of glucose [(16.7 ±3.5)mmol/L vs (5.1 ±1.1)mmol/L, t =13.9, P <0.01], glucagon [(158.5 ±50.2)ng/L vs (120.8 ±38.7)ng/L, t =2.5, P <0.05], epinephrine [(203.8 ±48.6)pg/ml vs (158.7 ±42.6)pg/ml, t =2.9, P <0.01], cortisol [(21.3 ±4.8)ng/ml vs (18.2 ±3.8)ng/ml, t =2.1, P <0.05], and HOMA-IR (10.9 ±2.6 vs 3.3 ±0.8 , t =12.3 , P <0.01 ) of diabetic rats were significantly increased .The glucose level [ ( 9.7 ±2.1 ) mmol/L vs ( 16.7 ± 3.5)mmol/L, t =7.0, P <0.01] was significantly improved after treated with insulin .The levels of epinephrine [(158.8 ±37.5 ) pg/ml vs (203.8 ±48.6)pg/ml, t =3.0, P <0.01], and cortisol [(15.7 ±4.2)ng/ml vs (21.3 ±4.8)ng/ml, t =3.5, P <0.01 ] were significantly improved after treated with fluoxetine .Conclusions Experimental stress increased the levels of glucose , en-docrine hormone , and insulin resistance of diabetic rats .Treatment with insulin improved the glucose level and treatment of fluoxetine improved the endocrine hormone level .

ObjectiveTo evaluate the effects on serum homocysteine(Hcy) level,vascular function and carotid intima-media thickness(IMT) in metformin-treated diabetic patients with or without supplementation with folate and Vitamin B12.MethodsA total of 100 newly diagnosed diabetic type 2patients were divided into two groups by random digits table with 50 cases each,90 patients completed study.Forty-seven participants (control group) received a 6-month course of metformin treatment,43 patients (treatment group) received mefformin,folate and Vitamin B12 treatment.The levels of serum Hcy,endothelin-1 (ET-1),carotid IMT,large or small arterial elasticity index (C1,C2),flow-mediated dilatation (FMD) of the brachial artery were evaluated before and after treatment.ResultsThe level of serum Hcy in control group significantly increased compared with before treatment[ (13.4 ± 2.7)μ mol/L vs.(11.1 ± 1.9)μ mol/L],hut the level of serum Hcy in treatment group significantly decreased compared with before treatment [ (9.2 ± 1.8 ) μ mol/L vs. ( 11.3 ± 2.0) μ mol/L ],there was significant difference(P ＜ 0.05 ).A beneficial effect was observed in the serum ET-1,FMD,carotid IMT and C2 in treatment group[ (20.0 ± 6.2)ng/L,( 15.8 ± 7.6)％,(0.8 ± 0.2) mm,(4.1 ± 2.1 ) ml/mm Hg ( 1 mm Hg =0.133 kPa) × 100 vs. (31.3 ±10.1 ) ng/L,(9.7 ± 4.5)％,( 1.1 ± 0.4) mm,(2.3 ± 1.0) ml/mm Hg × 100 ] (P ＜ 0.05).The levels of ET-1,FMD,carotid IMT and C2 after treatment in control group [ (24.8 ± 6.8) ng/L,( 12.9 ± 6.3 )％,(0.9 ± 0.3)mm,(3.0 ± 1.4) ml/mm Hg × 100] had significant difference compared with before treatment [ (30.6 ± 8.7)ng/L,(9.8 ± 4.6)％,( 1.0 ± 0.3) mm,(2.2 ± 0.9) ml/mm Hg × 100](P＜ 0.05).However,the results were improved significantly in treatment group than those in control group (P ＜0.05).In treatment group,significant correlation were detected between changes of Hcy and ET- 1 (r =0.43,P ＜ 0.05 ),carotid IMT(r =0.56,P ＜ 0.05),FMD (r =-0.54,P ＜ 0.05 ),C2 (r =-0.37,P ＜ 0.05 ).ConclusionsAdministration of folate and Vitamin B12 can reduce the levels of serum Hcy and ET-1 in metformin-treated type 2 diabetic patients,which exert beneficial effect on carotid IMT,FMD and C2.

Objective To investigate the toxic effect of immunosuppressive drugs on apoptosis of rat pancreatic islet cells in vitro and the protective action of Bcl-2.Methods Islet cells expressing Bcl-2 and the control islet cells were cultured at different concentrations of tacrolimus and the apoptosis rate of islet cells and insulin accumulation in the culture medium were detected after 48h.Results Low and high concentrations of tacrolimus induced the apoptosis of islet cells and decreased insulin secretion.The Bcl-2 inhibited the apoptosis of islet cells induced by tacrolimus and improved the insulin secretion.Conclusion Tacrolimus may directly damage to isolated rat islet cells and the expression of Bcl-2 can protect the cells from the damage of immunosuppressive drugs.

Objective To investigate the effects of antithyroid drug(ATD) on oxidative stress in Graves disease patients with and without infiltrative ophthalmopathy.Method The levels of superoxide dismutase(SOD),glutathione peroxidase(GPx) and catalase(CAT) in erythrocytes of Graves disease patients(18 with and 20 without infiltrative ophthalmopathy) were measured before and after treatment with ATD.Results Compared with the normal control group,the levels of SOD,GPx and CAT were significantly higher in Graves disease patients with and without infiltrative ophthalmopathy.After euthyroidism was achieved with the treatment of ATD,the levels of SOD,GPx and CAT were normalized in the patients without infiltrative ophthalmopathy,but oxidative stress was still present in the patients with infiltrative ophthalmopathy.Conclusion The oxidative stress is obvious in patients with Graves ophthalmopathy,which suggests that oxidative stress is involved in orbital inflammation.