Objective:To investigate the mechanism of inducing immune tolerance of autoimmune diabetic mice by insulin B chain gene vaccine Methods:Insulin B chain DNA vaccine was constructed by genetic engineering techniques DNA vaccine was injected into tibilias anterior muscle of C57BL/6 mice Mice were divided into four groups at random: group control (C), group diabetes (D), group treatment (T), prophylactic treatment (P) Serum was taken to determine insulin, the expression of bcl 2 mRNA was determined by dot blot dehydribation Results:①After prediabetic mice were injected by DNA vaccines, the ratio of incidence of diabetes was 30% in 2 w, 40% in 3 w and 4 w, which were significantly higher than that of groups D and T, and lower than that of group C The level of insulin showed no difference between group C and the non diabetic mice of group P In contrast to non diabetic mice, the change of pancreas of diabetic mice in group P was similar to group D ②AGV of bcl 2 mRNA in thymus cells, spleen cells, and blood monocytes of effective mice in group P showed no difference with group C, but markly lower than those of group D & T Conclusion:Insulin B chain DNA vaccine could recover the balance of the development and activity of thymus cells, spleen cells and lymphocyte in prediabetic mice

The serum level of 25-dihydroxyvitamin D3 was determined in patients with type 2 diabetes,diabetic nephropathy,and healthy objects..The results demonstrated that the serum 25-dihydroxyvitamin D3 level in type 2 diabetic patients with nephropathy was lower than that in the healthy objects or type 2 diabetes objects.Blood β2-M,total cholesterol,creatinine,24 h urinary albumin,and course of illness in type 2 diabetic patients with nephropathy have relationship with serum 25-dihydroxyvitamin D3.

Objective To investigate protective effects of pyrrolidine dithiocarbamate (PDTC) and mycophenolate mofetil (MMF) on kidneys of diabetic rats and the possible mechanism. Methods Thirty-four streptozotocin-induced diabetic rats were randomly divided into 4 groups: diabetic aminals without treatment (D group), diabetic rats treated with PDTC (P group), diabetic rats treated with MMF (M group), diabetic rats treated with combined PDTC and MMF (P+M group), and normal rats were considered as control group (C group). Blood glucose, blood urea nitrogen (BUN), serum creatinine (Scr), and 24 h urinary protein (24 h UP) were detected at the end of treatment lasting for 8 weeks. Kidneys were weighed in order to measure kidney weight/body weight ratio (KI) after rats were killed. Pathological changes in the kidneys were observed by light microscope and electron microscope. Expressions of interleukin-18 (IL-18), intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-α (TNF-α) in renal glomerulus were detected by immunohistochemical staining.Results Blood glucose, BUN, Scr, 24 h UP, KI were significantly higher in D, P, M and P+M groups than those in C group (all P＜0.05). BUN, Scr, 24 h UP, KI were significantly lower in P, M and P+M groups than those in D group (all P＜0.05). 24 h UP was reduced in P+M groups compared with P,M groups (P＜0.05). Pathological changes were attenuated in P,M and P+M groups compared with D group. Expressions of IL-18, ICAM-1, TNF-α in renal glomerulus were much higher in D group than those in C group (all P＜0.05) and were reduced in P, M and P+M groups compared with D group (all P＜0.05). Expressions of IL-18, ICAM-1, TNF-α in P+M group were lower than those in P group (all P＜0.05), while expressions of IL-18, TNF-α were lower than those in M group (both P＜0.05). Conclusion The protective effects on kidneys of diabetic rats produced by combined use of PDTC with MMF were much better than by the application of either PDTC or MMF alone. The mechanism appears to be related with suppressing expressions of inflammatory cytokines.

Objective To assess the efficacy of methylprednisolone (MP)pulse therapy in treatment of Hashimoto thyroiditis(HT) complicated with goiter.Methods 30 patients with HT complicated with goiter participated in the study and received MP pulse therapy.The patients had to be euthyroid for at least 3 months before the date of inclusion with plasma concentrations of thyroid hormones within their reference range.The goiter was still obvious and had no improvement.A dose of 250 mg MP was administered intravenously for seven consecutive days,and then treated with oral prednisone 10 mg,three times per day for 6 weeks with the dosage in each week gradually reduced at 5 mg to none.Ultrasonic was used to measure thyroid size before and after MP treatment.Results The treatment was successful at the end of the trial in all of the 30 patients receiving MP.The thyroid size from length,breadth,thickness to isthmus obviously decreased (P ＜ 0.05).The length of the left lobe was (57.42 ± 12.87) mm and (46.37 ± 7.67) mm (t =4.58) before and after treatment; The breadth of the left lobe was(26.68 ±7.71) mm and(22.21 ±6.09) mm(t =4.56) before and after treatment; The thickness of the left lobe was (27.18 ± 6.60) mm and (21.14 ± 5.67) mm(t =7.28) before and after treatment.The length of the right lobe was(58.17 ± 12.32)mm and(49.73 ±9.35) mm(t =3.84) before and after treatment; The breadth of the right lobe was (26.14 ± 7.37)mm and (23.00 ± 6.68) mm(t =3.29) before and after treatment ; The thickness of the right lobe was(27.57 ± 6.42)mm and(22.00 ±5.55)(t =5.88)before and after treatment.The isthmus before and after treatment was(9.94 ±4.15)mm and(6.19 ±2.57)mm(t =6.09).The recurrence rate was 17％ (5/30) after one year.Conclusions MP pulse therapy is an effective treatment for HT complicated with goiter.The recurrence rate is low.

The methylation status of GNAS1 gene in pseudohypoparathyroidism type Ib patients was detected by methylation-specific PGR technique. There was an abnormal methylation of 1A region in all seven PHPIb patients. Loss of exon 1A methylation (imprinting defect) seems to be the cause of PHPIb.

Objective To explore the effect of radiosurgery in combination of temozolomide therapy in vitro and in vivo on invasion and endocrine function of human glioma cell line U87-epidermal growth factor receptor and variant Ⅲ (EGFRvⅢ) and their xenografl.Methods (1) Human glioma U87-EGFRvⅢ cells were routinely cultured;CCK-8 was used to detect the effect of temozolomide at different concentrations on proliferation of U87-EGFRvⅢ cells,and the semi-inhibitory concentration (IC50) oftemozolomide was calculated;the U87-EGFRvⅢ cells were divided into control group,6 Gy radiotherapy group and 12 Gy radiotherapy group,and the expressions of invasion-related proteins and gene repair proteins in the three groups were detected by Western blotting after 0,6 and 12 Gy edge-dose irradiation,respectively;the U87-EGFRvⅢ cells were divided into control group,6 Gy radiotherapy group and 6 Gy radiation combined with temozolomide treatment group (5 mmol/L temozolomide was added after 6 Gy irradiation),and 24 h after each treatment,Western blotting was used to detect the expressions of invasion related proteins and gene repair related proteins in the three groups.(2) Nude mouse transplantation intracranial glioma models were established with cell suspension of the control group,6 Gy radiotherapy group and 6 Gy radiation combined with temozolomide treatment group;7,14,21 d after transplantation,biolumineseence imaging (BLI) was employed to detect luciferase expression in nude micein vivo and measurement of transplanted tumors was performed;survival curve and body mass curve were drew in nude mice;immunohistochemical staining was used to detect the expressions of EGFRvⅢ,epidermal growth factor receptor (EGFR),matrix metalloproteinase (MMP)-9,vascular endothelial growth factor (VEGF) and Ki67 in intracranial transplanted gliomas.Results (1) The IC50 oftemozolomide on U87-EGFRvⅢ cells was 5 mmol/L;as compared with those in the control group,the expressions of EGFRvⅢ,EGFR,MMP-2,MMP-9,VEGF,MGMT,AKT-1,β-catenin and Ku70 significantly increased in the cells of the 6 Gy radiotherapy group,while the expressions of EGFRvⅢ,EGFR,MMP-2,MMP-9,VEGF,MGMT,AKT-1,β-catenin and Ku70 significantly decreased in the cells of the 12 Gy radiotherapy group as compared with those in the 6 Gy radiotherapy group (P＜0.05);as compared with those in the 6 Gy radiotherapy group,the expressions of EGFRvⅢ,EGFR,MMP-2,MMP-9,VEGF,MGMT,AKT-1,β-catenin and Ku70 in cells of the 6 Gy radiotherapy combined with temozolomide group were statistically decreased (P＜0.05).(2) As compared with mice in the control group and 6 Gy radiotherapy group,mice in the 6 Gy radiotherapy combined with temozolomide group had significantly lower cell tumorigenicity,significantly higher survival rate and body mass 21 d after transplantation,and significantly smaller volume of xenograft tumors (P＜0.05);the expressions of EGFRvⅢ,EGFR,MMP-9,VEGF and Ki67 in tumor tissues of nude mice in the 6 Gy radiotherapy group were significantly decreased as compared with those in the control group;as compared with those in the 6 Gy radiotherapy group,the expressions of EGFRvⅢ,EGFR,MMP-9,VEGF and Ki67 in tumor tissues of mice in the 6 Gy radiotherapy group were significantly decreased (P＜0.05).Conclusion The combined treatment of radiosurgery and temozolomide can more obviously inhibit the invasiveness of U87-EGFRvⅢ cells and it exerts a stronger inhibitory effect on tumorigenicity of U87-EGFRvⅢ cells in brain of nude rats.

Objective To evaluate the effect of ultra short-term low-dose gamma knife radiation on invasive properties of glioma cells at the genetic level.Methods Malignant glioma cell lines U87 and U251 were treated with gamma knife radiation,respectively,with a marginal doses of 0,4,6,8 and 10 Gy (all to the 80％ isodose line).Real time-PCR was used to measure the mRNA expressions of AKT-1,AKT-2,β-catenin and TCF-4 at 30 min after radiation.Results The expressions of AKT-1,AKT-2,β-catenin,TCF-4 in glioma cell lines after low-dose gamma knife radiation with a marginal dose of 6-8 Gy were significantly increased as compared with those after 0 Gy (P＜0.05); reduced expressions ofAKT-1,AKT-2,β-catenin and TCF-4 after a marginal dose of 10 Gy were found as compared with those after 0 Gy (P＜0.05); the β-catenin and AKT-1 mRNA expressions in U87 cells and TCF-4,AKT-1 and AKT-2 mRNA expressions in U251 cells enjoyed the most obvious increase at a marginal dose of 6 Gy; the TCF-4 and AKT-2 mRNA expressions in U87 cells and β-catenin mRNA expression in U251 cells enjoyed the most obvious increase at a marginal dose of 8 Gy.Conclusion Low-dose gamma knife radiation (6-8 Gy) could increase the β-catenin,TCF-4,AKT-1 and AKT-2 expressions,while high dose (10 Gy) could inhibit these effects in a short time period of 30 min.

Although high-efficiency targeted delivery is investigated for years, the efficiency of tumor targeting seems still a hard core to smash. To overcome this problem, we design a three-step delivery strategy based on streptavidin-biotin interaction with the help of c(RGDfK), magnetic fields and lasers. The ultrasmall superparamagnetic iron oxide nanoparticles (USIONPs) modified with c(RGDfK) and biotin are delivered at step 1, followed by streptavidin and the doxorubicin (Dox) loaded nanosystems conjugated with biotin at steps 2 and 3, respectively. The delivery systems were proved to be efficient on A549 cells. The co-localization of signal for each step revealed the targeting mechanism. The external magnetic field could further amplify the endocytosis of USPIONs based on c(RGDfK), and magnify the uptake distinctions among different test groups. Based on photoacoustic imaging, laser-heating treatment could enhance the permeability of tumor venous blood vessels and change the insufficient blood flow in cancer. Then, it was noticed that only three-step delivery with laser-heating and magnetic fields realized the highest tumor distribution of nanosystem. Finally, the magnetism/laser-auxiliary cascaded delivery exhibited the best antitumor efficacy. Generally, this study demonstrated the necessity of combining physical, biological and chemical means of targeting.