Objective To study the effects of citrullinated vimentin (cVim) on the maturation and immunologic function of dendritic cells (DCs) from rheumatoid arthritis (RA) peripheral blood.Methods In the present study,mononuclear cells were isolated from the peripheral blood of patients with RA and cultivated in media containing GM-CSF and IL-4 to generate immature DCs (imDCs).The imDCs generated were stimulated with citrullinated vimentin and vimentin.LPS was used as the positive control and PBS was used as the negative control.The expression of surface molecules on the DCs,such as CD14,CD80,CD83,CD86,MHC Ⅰ and MHC Ⅱ were analyzed with FACS.The capability of the stimulatory activity of the DCs on allogeneic T cells in mixed reaction was tested by MTS.t-test was used for statistical analysis.Results Compared to untreated DCs,DCs treated with LPS increased the expression levels of MHC Ⅱ,CD80,CD83 and CD86 (1.07±0.14,1.25±0.13,1.90±1.08,2.44±0.65,P＜0.05),while cVim increased the expression levels of MHC Ⅱ ( 1.18±0.09,P＜0.05) and CD83 ( 1.97±0.99,P＜0.01 ),and Vim decreased the expression levels of CD80 (0.82±0.18,P＜0.01 ).It was demonstrated that the expression levels of MHC Ⅱ on DCs pulsed with cVim were significantly higher than that of the DCs with LPS,but the expression levels of CD80 and CD86 were not significantly different.The expression levels of MHC Ⅱ and CD83 on DCs pulsed with cVim were significantly higher than that of the DCs with Vim.The mixed lymphocyte reaction showed that the DCs induced by LPS and cVim trigerred the proli-feration of allogenic T cells obviously.Conclusion This result suggests that cVim could promote the phenotypic maturation of DCs and increase the expression of costimulatory molecules.

Adolescent ; Adult ; Aged ; Facial Nerve ; anatomy & histology ; pathology ; Female ; Humans ; Male ; Microsurgery ; Middle Aged ; Parotid Neoplasms ; surgery ; Young Adult

To prove the influence of protein isoaspartyl-methyltransferase ( PIMT ) on the cell apoptosis of fibroblast-like synoviocytes of RA.Methods: The expression vector of PIMT was constructed and transfected in to the RA-FLS, the impact of PIMT on the cell apoptosis of RA-FLS was observed by overexpressing the vector of PIMT.Results:The mRNA and protein level of PIMT in RA-FLS was increased after transfected the vector of PIMT into RA-FLS;compared with the normal cultured RA-FLS and the RA-FLS transfected with the empty vectors ,the cell apoptosis level was also increased.Conclusion:The decreased expression level of PIMT in RA-FLS is an important reason for reduce apoptosis of RA-FLS,and PIMT can affect the imbalance of proliferation/ap-optosis in the RA-FLS.

Objective To study the injury and protective action of drugs on neurons, the model of glutamate excitotoxicity on primary cultured hippocampual neurons from new born rats were( set up. Methods)By use of trypan blue dye staning and testing the lactate dehydrogenase leakage from cultured neurons, to investigate the neuron survival rate. Results We found the injury of neurons was related with the concentration of glutamate. NMDAR non-competitive antagonist —MK-801 could protect the glutamate excitotoxic damage on neurons. Conclusion The glutamate results in neuron injury through NMDAR; the model of neuron culture was sufficient for glutamate-induced excitotoxicity.

Objective:To prove the effect of PADI-4 gene in the development of rheumatoid arthritis.Methods:Four SiRNA sequences were designed for PADI-4 gene,and the SiRNAs were cloned into blank pSiRNA-hH1neo G2 vectors.The vectors were transformed into GT116 E.coli competent cells.By white-blue selection system,the right vectors were gotten.After transfection into HL-60 cells,the cells were collected on 3,5,7,10 and 14 day,the levels of PADI-4 mRNA were detected by Real-time PCR.Results:Digestion by Acc 65Ⅰand Hind Ⅲ,the recombinant expressive vector of RNA interference was obtained successfully.The PADI-4 mRNA generated by the cells transfected with the vector of SiRNAs were reduced,and the level was not change in normal cells.Conclusion:The recombinant expressive vector of RNA interference is obtained successfully and the recombinant expressive vector can affect expression of PADI-4 gene in HL-60 cells.

OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

China ; Congresses as Topic ; Fatty Liver ; Humans

Objective To explore the association between fibrinogen gene polymorphism(rs1049636) and serum γ′ fibrinogen level and ischemic stroke (IS) .Methods 421 IS patients and 421 age‐and gender‐ matched healthy controls ,including 283 males and 138 females ,were recruited in this assay .The plasma γ′fibrinogen concentration was measured by enzyme‐linked immunosor‐bent assay (ELISA) .Fibrinogen gene polymorphism(rs1049636) were genotyped by using PCR‐LDR assay .Results γ′fibrinogen concentrations in IS patients[(159 .4 ± 97 .4)U/dL] were significantly higher than that in control group[(114 .2 ± 73 .0)U/dL] with statistically significant difference(P<0 .001) .Single nucleotide polymorphism(SNP) analysis showed that rs1049636 C allele was significantly associated withγ′fibrinogen level ,but not associated with increased risk of IS(P=0 .077) .Conclusion An associ‐ation between increasedγ′fibrinogen level and IS existed in Chinese Han population .However ,no association between rs1049636 C allele and IS risk was observed in our study .

Objective To investigate the relationship between gene polymorphism of transcripion factor 7-like 2 (TCF7L2) at positions rs290487, rs11196205, rs11196218 and gestational diabetes mellitus (GDM) in Chinese women.Methods In 1140 unrelated pregnant Northern Chinese women (335 women with GDM, 158 gestational cases with impaired glucose tolerance and 647 pregnant non-diabetic controls) ,three single nucleotide polymorphisms (rs290487, rs11196205, and rs11196218) in the TCF7L2 gene were genotyped using ligase detection reaction (LDR).In the present study, cases with GDM and impaired glucose tolerance (IGT) were indistinguishable clinically and biochemically, and were combined into case group.Results The frequency of C allele of rs290487 was 41.6% in case group, being significantly higher than that in control group (36.3%, P=0.012).There was significant difference in the frequency of CC genotype between case group and control group (18.7% vs 14.0%, P=0.033).Compared with T allele carriers, CC genotype carriers had a 1.418-fold increased risk of GDM (95% CI 1.028-1.955).After adjusting for age, body mass index, family history of diabetes,systolic blood pressure,and diastolic blood pressure, pregnant women with CC genotype carriers of rs290487 were more prone to hyperglycemia compared with the T allele carriers (OR 1.518, 95% CI 1.064-2.166).Conclusions The TCF7L2 rs290487 variant may contribute to the genetic predisposition to GDM.CC genotype is likely to be associated with an increased risk of GDM in the pregnant Chinese women.

Objective:To explore the status of adolescents'empathy and its related factors.Methods:Totally 1460 students from part of middle schools and vocational schools in Huainan City and Wenzhou City were surveyed,1159 (613 males,546 females) valid samples obtained.Subjects'empathy,family function and class environment were separately assessed with Interpersonal Reactivity Index-C(IRI-C),Family Assessment Device(FAD) and the questionnaire of My Class.Results:IRI-C scores were higher in gifts than in boys [total empathy,(55-± 12) vs.(51 ± 11),P ＜ 0.05].Vocational school students' IRI-C scores were lower than junior school students' [total empathy,(50 ± 11) vs.(54 ± 11),P ＜ 0.001],and lower than high school students' [total empathy,(50 ± 11) vs.(54 ± 11),P ＜ 0.05].Competition scores and classmates scores were positively associated with emotional empathy scores (β =0.11,0.13,P ＜ 0.001).The role scores were positively associated with the scores of emotional empathy(β =0.14,P ＜ 0.001).Problem solving scores were negatively associated with cognitive empathy and total empathy scores (β =-0.22,-5.20,P ＜ 0.001).Classmates scores were positively associated with cognitive empathy and total empathy scores (β =0.18,5.76,P ＜ 0.001).Conclusion:It suggests that girls' empathy may be better than boys'.Vocational school students'empathy may be worse than junior school students'and high school students Good relationship with classmates,efficient solutions to the problems and moderate competition may be favorable factors for adolescents' empathy.