Objective To observe the function and survival of parathyroid cells cultured under simulated microgravity condition after cotransplanation of syngeneic allogeneic testicular sertoli cells.Methods Parathyroid cells in SD rats were assigned to flask-culture or bioreactor-culture.Allogeneic testicular sertoli cells in SD rats were cultured by using routine method.The recipients of hypoparathyroidism Wistar rat models were divided into 3 groups randomly:group A,receiving parathyroid cells(cultured with routine method)transplantation only;group B,receiving parathyroid cells and allogeneic testicular sertoli cells(cultured with routine method)transplantation;group C,receiving parathyroid cells(cultured under simulated microgravity condition)and allogeneic testicular sertoli cells transplantation.Allograft survival,change in cell components,apoptosis of infiltrative lymphocytes and parathyroid cells function were analyzed after transplantation respectively.Results The average survival time in group A,B and C was(17.3±1.6),(43.2±2.4)and (52.5±1.5)days,respectively.There was significant difference among group (P<0.05).In group C,parathyroid cells with strong secreting function grew into scaffold materials and adhered to them.FasL-expressing testicular cells and apoptotic lymphocytes were quite evident between allograft and kidney parenchyma.Conclusion Parathyroid cell cultured under simulated microgravity condition enhances its survival and function after cotransplanation of allogeneic testicular sertoli cell with immune privilege.

Objective To observe the antioxidant effects of 99Tc-MDP on the brain of aged mice induced by D-galactose.Methods A total of 48 healthy female Kunming mice were divided into 6 groups by random number table method:normal control group,model group,vitamin E (Vit E) group,groups treated with low,middle and high dosages of 99Tc-MDP.Except the normal control group,mice of each group were injected with 10％ D-galactose saline subcutaneously in the neck for 42 d.At the same time,the 99Tc-MDP groups were given different dosages of 99Tc-MDP (1.75× 10-5,3.50× 10-5,0.70× 10-4 μg/g according to the body mass) respectively into abdominal cavity twice a day.Vit E group was given Vit E by intragastric administration from the first day.The model group was injected with saline every day.After behavioral testing,mice serum samples and brain tissue samples were collected for the determination of superoxide dismutase (SOD),malondialdehyde (MDA),glutathione peroxidase (GSH-Px) and monoamine oxidase (MAO)levels.Then the mice were sacrificed and the brain tissue was taken for HE staining.The independent-sample t test was used for data analysis.Results The mice model was established successfully.SOD levels in brain tissue of groups injected with low,middle,high dosages of 99Tc-MDP,Vit E group were (19.06± 6.44),(20.41±4.02),(22.24±3.76),(24.71±5.09) U/mgprot,respectively,all of which were higher than that of the model group((11.32±2.90) U/mgprot;t=3.099 6-6.504 6,P＜0.05 or ＜0.01).There were similar results for GSH-Px (t =2.214 1-4.145 7,P＜0.05 or ＜0.01).MDA and MAO levels in brain tissue of 99Tc-MDP groups were lower than those of the model group (t =2.140 3-3.057 8,all P＜0.05).Compared to normal control group,the hippocampus in model group showed reduced cell number and layers,disordered structure,with part of the cells in smaller volume and abnormal nuclear shape.In Vit E group and the three 99Tc-MDP groups,no significant change of neuron was observed compared with normal control group,the degeneration and necrosis of hippocampal cells were mild compared with model group,the cell number and morphology were normal,and the structures were clear.Conclusion 99Tc-MDP may increase the activities of SOD,GSH-Px and reduce the levels of MDA and MAO in brain tissue of aged mice,thus it may be helpful in delaying the brain aging.

Objective To investigate the association between the genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) and ulcerative colitis (UC) of Han ethnic population in Zhejiang, China. Methods Two hundred and seventy-four consecutive patients with UC and 726 healthy controls (HC) were studied. The genetic polymorphisms of MTHFR (C677T and A1298C) were genotyped using PCR-RELP methods. Results The frequencies of variant allele and genotype in MTHFR A1298Cgene were higher in UC patients than in the HC (35.77% vs 29. 96%, P =0. 013; 52. 19% vs 44. 90%,P=0.039; respectively). However, there were no significant discrepancies of the allele and genotype frequencies in the MTHFR C677T gene between the UC patients and the HC (P ＞ 0. 05 ). In addition, the MTHFR 677Tr homozygote, T allele and 677CT/1298AC compound genotype were more prevalent in patients with extensive colitis than in those with distal colitis (37. 66% vs 14. 72% ,P = 0. 0002; 49. 35% vs 32.99% ,P =0. 0004; 29. 87% vs 15.23% ,P =0. 006; respectively). Furthermore,the variant allele in the MTHFR A1298C gene (C) in severe UC patients was significantly lower than in mild and moderate UC patients (18.97% vs 33. 88% ,P =0. 022). Conclusion The genetic polymorphisms of MTHFR C677T and A1298C are obviously associated with Han ethnic population with UC in Zhejiang province.

Aim To investigate the effects of starfish saponins(Sfs) on insulin signaling pathway in orotic acid-induced NAFLD rats.Methods 1% orotic acid was used to establish NAFLD model in male Wistar rats for six weeks.The NAFLD rats were randomly divided into two groups(eight rats in each group) and then fed with the corresponding diets: Model group(1% orotic acid)and Sfs group(1% orotic acid containing 0.04% starfish saponins).After starfish saponins feeding for 8 weeks, hepatic lipids content, liver function indices and relevant protein expression in muscle insulin signaling pathway were measured.Results Compared with model group, starfish saponins reduced hepatic lipids content and improved liver functions.In addition, it effectively ameliorated insulin resistance by improving insulin signaling pathway and improved glucose uptake in muscle.Conclusion The amelioration effect of starfish saponins on impaired insulin signaling pathway in muscle is observed in orotic acid-induced NAFLD rats.

Objective:To Studythe extracellular matrix metalloproteinase inducer (CD147) changes in patients with systemic lupus erythematosus (SLE), and to study thecorrelation of CD147 level and athero-sclerosis in SLE.Methods:Eighty patients with SLE in total were divided into intimal thickening group, (24 cases, group A) and normal intimal group (56 cases, group B) according to carotid intimamedia thickness (IMT) checked by carotid ultrasonography. In addition, their age, bodymass index, blood pressure, seral total cholesterol (TC), high density liptein cholestero (HDL-C), low density lipoprotein cholestero (LDL-C), triglyceride (TG), urinary protein quantitative test(24 h), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), anticardiolipin antibody, serum creatinine level, course of the disease, and treatment regimens were collected. Thirty-five healthy people were set as the control group (group C). The levels of serum CD147 were measured by enzyme-linked immuno sorbent assay (ELISA) in 3 groups. The correlation between the serum CD147 level of SLE patients and atherosclerosis was analyzed. The statistical analysis was carried out with independent t-test, chi-square test, analysis of variance, Spearman correlation and Logistic regression. Results:① Levels of serum CD147 in group A [ (238±30) pg/ml] were significantly higher than those in group B [(198±30) pg/ml] and group C [(150±26) pg/ml, F=67.908, P<0.01]. ② Body mass index, hypertensive ratio,total blood cholesterol,urine protein quantitative test (24 h), systemic lupus erythematosus disease activity index (SLEDAI), serum CD147 index in group A were significantly higher than those in group B ( P<0.05). ③ In Logistic regression analysis, serum CD147 [ OR (95% CI)=1.039(1.014, 1.065), P<0.05], urine protein quantitative (24 h) [ OR (95% CI)=2.598(1.033, 6.534), P<0.05] were independently relevant factors affecting carotid artery IMT. Conclusion:Serum CD147 is an independent risk factor for carotid intimamedia thickness in SLE patients.

ObjectiveTo investigate the characteristics of trace elements and amino acid metabolism in the blood of patients with hepatocellular carcinoma (HCC) and to guide clinical treatment. MethodsThis study included 125 untreated patients with primary HCC in Tangshan Maternal and Child Health Hospital and Tangshan Guzhi District Hospital from March to November, 2013, as well as 50 healthy controls who underwent physical examination in Tangshan Maternal and Child Health Hospital. Blood trace elements were measured by atomic absorption spectrophotometry, and the levels of amino acids were determined by an amino acid analyzer. Comparison of continuous data was made by t test. ResultsCompared with the controls, the HCC patients had a significantly higher Cu level (t=6.30, P＜0.05) and significantly lower Zn, Fe, and Mg levels (t=10.15, 3.98, and 2.25, all P＜0.05). The HCC patients showed significant increases in tyrosine, phenylalanine, and methionine levels (t=10.10, 13.46, and 2.34, all P＜0.05) and significant decreases in valine, isoleucine, leucine, and arginine levels (t=3.65, 2.98, 5.67, and 8.35, all P＜0.05). ConclusionHCC patients show changes in various amino acids and trace elements in blood. Measurement of trace elements and amino acids is of certain significance for evaluating HCC patients′ condition and guiding clinical treatment.

Objective To assess the expression and discuss the clinical significance of CD4+CD25+ regulatory T cells and Thl7 cells in peripheral blood of patients with gastric carcinoma. Methods 30 patients with gastric carcinoma in our hospital were randomly selected as the gastric carcinoma group. 30 people who checked up their health condition in our hospital during the same period were selected as the normal group. The expression of CD4, CD25, Foxp3 and IL-17 in peripheral blood was assessed in both groups. The amount and proportion of CD4+CD25+ regulatory T cells and Thl7 cells were analyzed. Results The expression of CD4+CD25+ regulatory T cells and Thl7 cells in gastric carcinoma group increased compared with that in normal group (P＜0. 05). The expression of CD4+CD25+ regulatory T cells in peripheral blood of patients with advanced gastric carcinoma, gastric carcinoma in T3+T4 and gastric carcinoma with lymph node metastasis was higher than that in patients with early gastric carcinoma, gastric carcinoma in T1+T2 and gastric carci noma without lymph node metastasis respectively (P＜0. 05). No significant difference was noticed in the expression of Th17 cells in patients with gastric carcinoma in different stages, depth of infiltration or with lymph node metastasis (P＞ 0. 05). Conclusion The expression of both CD4+CD25+ regulatory T cells and Thl7 cells in peripheral blood of patients with gastric carcinoma increased significantly. It was also suggested the progression of gastric carcinoma was associated with CD4+CD25+ regulatory T cells, but not associated with Th17 cells statistically.

Objective:To analyze the serum levels of integrin-associated proteins (CD47) in patients with rheumatoid arthritis (RA), and to explore its association with disease activity and bone destruction in RA.Methods:Serum and clinical data were collected from 65 RA patients and 25 healthy subjects. RA patients were grouped into low, moderate, and high bone erosion groups according to 7-joint ultrasonography score (US7). The levels of serum CD47, thrombospondin-1 (TSP-1) and receptor activator of nuclear factor-κB ligand (RANKL) were measured by enzyme-linked immunosorbnent assay (ELISA) in patients with RA and healthy subjects. The statistical analysis was carried out with independent t-test, analysis of variance, nonparametric rank sum test, pearson or Spearman correlation and logistic regression. Results:① The Serum levels of CD47, TSP-1, and RANKL were higher in the RA group than in the healthy controls ( P<0.01). ② In RA patients, serum CD47 level was positively correlated with disease course ( r=0.301, P<0.05), C-reactionprotein (CRP)( r=0.316, P<0.05), number of tender joints (TJC) ( r=0.254, P<0.05), number of swollen joints (SJC) ( r=0.316, P<0.05), disease activity score in 28 joints (DAS28) ( r=0.255, P<0.05), RANKL ( r=0.252, P<0.05) and TSP-1 ( r=0.260, P<0.05). Serum TSP-1 level was positively correlated with CRP ( r=0.299, P<0.05), TJC ( r=0.335, P<0.01), DAS28 ( r=0.315, P<0.05), RANKL ( r=0.305, P<0.05). ③ The disease course [ OR(95% CI)=1.048(1.033, 1.017)] and TSP-1 [ OR(95% CI)=1.013(1.000, 1.026)] were independently relevant factors affecting bone destruction. Conclusion:CD47 levels is significantly higher in RA patients than in healthy controls, and is associated with disease activity and bone destruction. CD47 may be involved in the bone destruction process of RA by acting on TSP-1.

Objective:To investigate the effect of tofacitinib on early atherosclerosis of patients with systemic lupus erythematosus and explore the possible relationship between lupus nephritis and early atherosclerosis of systemic lupus erythematosus.Methods:Sixteen 8-week-old female MRL/lpr mice with a body weight of 20~25 g were selected and randomly divided into the treatment group and placebo group, with 8 mice in each group. The treatment group diluted tofacitinib by normal saline, and given at a dose of 10 mg·kg -1·d -1, and the placebo group (starch tablets) administered the medication in the same way as the treatment group for a total of 8 weeks. The ELISA method was applied to detect serum anti-dsDNA antibody levels in the two groups of mice. Bradford method protein concentration was used to determine the level of urine protein in mice. Automatic biochemical analyzer was used to detect blood lipids, urea nitrogen, serum creatinine, complement C3, complement C4 levels. Western blotting was used to determine the protein expression levels of monocyte chemoattractant protein-1 (MCP-1), non-receptor protein tyrosine kinase family 1 (JAK1), signal transducer and activator of transcription 1 (STAT1) and signal transducer and activator of transcription 2 (STAT2) in aortic and kidney tissues. After the aortic arch section were prepared, oil red O was used to stain the sections, and the vascular plaque area and intimal thickness were evaluated by ImageJ software. The kidneys were dissected and stained with HE, and the active lesions of lupus nephritis were evaluated using the glomerular activity scoring system. SPSS 23.0 software was used for statistical analysis, in which the between-group comparison was performed using two independent samples t-test, and the correlation analysis was performed using the Spearman method. Results:①The serum anti-dsDNA antibody expression level in the treatment group [(5.2±1.0) U/ml] was lower than that in the placebo group [(6.9±1.2) U/ml], ( Z=-3.07, P=0.008), and the levels of complement C3 and complement C4 were higher than those in the placebo group [(293±10) mg/L vs. (260±19) mg/L, Z=2.72, P=0.017]; (16±6) mg/L vs. (8±9) mg/L, Z=3.78, P=0.006]. There was no significant difference in serum BUN and Scr between the treatment group and the placebo group [(10.6±0.7) mmol/L vs. (11.5±1.1) mmol/L, Z=-1.96, P=0.071; (17±5) μmol/L vs. (22±6) μmol/L, Z=-1.79, P=0.095]. ② Compared with the placebo group, the levels of LDL, TC and TG in the treatment group decreased [(0.83±0.15) mmol/L vs. (1.08±1.05) mmol/L, Z=-3.95, P=0.001; (2.90±0.08) mmol/L vs. (1.81±0.97) mmol/L, Z=-5.17, P=0.001; (1.10±0.08) mmol/L vs. (1.60±0.42) mmol/L, Z=-3.23, P=0.013], and HDL level increased [(2.02±0.99) mmol/L vs. (1.81±0.97) mmol/L, Z=4.42, P=0.001]. ③ Compared with the placebo group, the levels of aortic MCP-1, JAK1, STAT1 and STAT2 in the treatment group were reduced [(0.17±0.30) vs. (0.23±0.05), Z=-3.06, P=0.009; (0.83±0.09) vs. (1.05±0.19), Z=-3.07, P=0.008; (0.77±0.07) vs. (0.94±0.13), Z=-2.83, P=0.014; (0.70±0.07) vs. (0.82±0.09), Z=-2.83, P=0.013], the aortic plaque area and aortic intimal thickness were lower than those in the placebo group [(12±31) μm 2vs. (1 242±1 101) μm 2, Z=-3.12, P=0.016; (63±7) μm vs. (82.10±8.06) μm, Z=-5.13, P<0.001]. ④ Compared with the placebo group, the urine protein level and glomerulonephritis activity score in the treatment group were decreased [(0.08±0.03) mg/mL vs. (0.20±0.11) mg/mL, Z=-3.08, P=0.015; (1.79±0.38) vs. (2.79±0.14) points, Z=-7.08, P<0.001)], and renal tissue MCP-1, JAK1, STAT1.Compared with the placebo group, STAT2 levels were reduced [(0.364±0.040) vs. (0.425±0.021), Z=-3.85, P=0.003; (0.689±0.074) vs. (0.838±0.068), Z=-4.19, P=0.001; (0.508±0.070) vs. (0.646±0.019), Z=-2.85, P=0.015; (0.618±0.062) vs. (0.740±0.101), Z=-2.94, P=0.013. ⑤ The glomerular mobility scores of the two groups were positively correlated with LDL, TCHO, TG, aortic plaque area and aortic intimal thickness ( r=0.51, P=0.043; r=0.79, P<0.001; r=0.64, P=0.008; r=0.82, P<0.001; r=0.74, P=0.001), and negatively correlated with HDL ( r=-0.53, P=0.036). The urine protein levels in the two groups were positively correlated with LDL, TC, TG, aortic plaque area and aortic intimal thickness ( r=0.67, P=0.004; r=0.68, P=0.004; r=0.53, P=0.033; r=0.80, P<0.001; r=0.74, P=0.001), and negatively correlated with HDL ( r=-0.57, P=0.021). Conclusion:The severity of lupus nephritis is correlated with atherosclerosis and dyslipidemia in the early stage of systemic lupus erythematosus. Tofacitinib may reduce the degree of early arteriosclerosis and lupus nephritis in MRL/LPR mice, and reduce blood lipid levels, which may be effective in improving the prognosis of SLE and improving the survival rate of patients.

Objective To investigate the relationship between gene polymorphisms of T cell immunoglobulin domain and mucin domain protein-3 (Tim-3) and ulcerative colitis (UC) in Han nationality of Zhejiang.Methods A total of 391 UC patients and 573 healthy controls were recruited.Two single nucleotide polymorphisms (SPNs) of Tim-3 (rs1036199 and rs10515746) were examined by the improved multiple ligase detection reaction technique.Chi-square test or Fisher's exact test was performed to analyze the differences in the distribution of Tim-3 gene polymorphisms and its influence on the location and severity.Haploview 4.2 software was used to analyze linkage disequilibrium (LD) and haplotype.Results The frequencies of genotype CA+AA and mutant allele A of rs10515746 in UC were lower than those in healthy controls (1.79％,7/391 vs 4.19％,24/573;0.90％,7/782 vs 2.18％,25/1 146;x2=4.295 and 4.712,P=0.038 and 0.030).However,there was no significant differences in frequencies of genotype CA+ CC and mutant allele C of gene rs1036199 between UC patients and the healthy controls (1.79％,7/891 vs 8.49％,20/578;0.90％,7/782 vs 1.74％,20/1 146;both P＞0.05).The frequencies of genotype CA+AA and mutant allele A of rs10515746 in mild and moderate UC patients were both higher than those in severe UC patients (2.87 ％,7/244 vs 0;1.43 ％,7/488 vs 0),and the differences were statistically significant (Fisher's exact test,P=0.049 and 0.048).The analysis for LD indicated that rs1036199andrs10515746 were closeLD (D'=0.92,r2=0.72).Furthermore,the frequency of haplotype CA formed by the mutant alleles C and A of these two SNPs was lower in UC patients than that in healthy controls (0.64％,5/782 vs 1.74％,20/1 146),and the difference was statistically significant (x2 =4.441,P=0.035).Conclusions Tim-3 (rs10515746) gene mutation may not only decrease the incidence,but also reduce the severity of UC.Moreover,the haplotype CA formed by the mutant alleles of rs1036199 and rs10515746 may also reduce the incidence of UC.