ObjectiveTo investigate the effects of over expression of miR-34a on cellular proliferation and migration in bladder cancer cell line J82 by targeting Notchl.MethodsmiR-34a was predicted as a putative gene which can target Notchl through bioinformatics analysis,qRT-PCR and Western blot were performed to measure the expression levels of Notchl and miR-34a in invasive transitional cell carcinoma of bladder (TCCB) tissues and J82 cells transfected with miR-34a.Luciferase assay was employed to determine if miR-34a could target Notchl through binding to the 3'-untranslated region (3'UTR) of Notchl mRNA.J82 cells were transfected with pcDNA3.0-miR-34a or pcDNA3.0 control plasmid.MTS colorimetry was used to evaluate the effect of miR-34a on cell proliferation.The effect of miR-34a on cell migration was assessed by transwell migration assay.ResultsThe expression level of miR-34 in invasive TCCB tissues was lower than in adjacent bladder tissues (0.016(0.018) vs 0.042 (0.059),N =16; P =0.0006).On the contrary,the average levels of Notchl mRNA and protein were higher in tumors than in adjacent bladder tissues (2.765(2.156) vs 2.312(1.365),N =16; P =0.0025 and 0.857 ±0.197 vs 0.648 ±0.171 ;P ＜0.0001 ).After the transfection of miR-34a,the expressive level of miR-34a in J82 was highly induced ( (2.408 ±0.789) × 10-4 vs(0.153 ±0.029) × 10-4; P =0.0026).However,the expressive levels of Notchl mRNA and protein were obviously decreased (3.001 ± 0.106 vs 4.998 ± 1.053 ; P =0.0308 and 0.747 ± 0.050 vs 0.988 ± 0.102 ; P =0.0215 ).The results of luciferase assay showed that firefly activity was highly dimished (0.422 ± 0.028 vs 2.392 ± 0.148 ; P ＜ 0.0001 ).Cellular proliferation was inhibited after the transfection of miR-34a in J82 (P ＜ 0.0001 ).Moreover,number of migration cells of J82 was significantly reduced after the ectopic expression of miR-34a ( 179.3 ± 21.02 vs 269.7 ± 23.71 ; P =0.0078 ).ConclusionsmiR-34a inhibits the cellular proliferation and migration of bladder cancer cell line J82 via binding to the 3UTR of Notchl mRNA.

Objective To observe the short-term,long-term clinical results and complications in refractory juvenile idiopathic arthritis (JIA) treating with TNF-α inhibitors,and to compare 2 evaluation systems.Methods A retrospective review of 52 cases of patients with refractory JIA in Shenzhen Children's Hospital was performed.With reference to International Leagne of Associations for Rheumatology(ILAR)2001 diagnostic criteria,the patients were divided into 4 groups:26 polyarticular JIA patients,14 systemic JIA patients,9 oligoarticular JIA patients and 3 other types of patients.The children with JIA were based on the conventional treatment such as Methotrexate,combination of TNF-α inhibitors treatment.The short-term and long-term clinical outcomes were evaluated and compared with American College of Rheumatology (ACR) and Juvenile Arthritis Disease Activity Score (JADAS).Complications in each group were recorded.Results (1) Short-term outcome assessment:ACR 50 were achieved in 69.2％ of the polyarticular JIA,66.7％ in oligoarticular and 35.7％ in systemic JIA patients on the third month;and by the time of the sixth month it reached to 73.0％ in polyarticular JIA,77.7％ in oligoarticular JIA and 14.3％ in systemic JIA patients on the sixth month.Significant improvement of JADAS after the treatment was observed in each type of JIA,and the differences were statistically significant(all P ＜ 0.05).(2) Long-term outcome assessment:except for the cases missing follow-ups and withdrawal cases,46 patients were able to complete 2 years assessments:according to ACR,effective rate was 84.0％ in polyarticular JIA (21/25 cases),50.0％ in oligoarticular JIA (4/8 cases) and 7.7％ in systemic (1/13 cases) JIA patients;JADAS was significantly decreased in polyarticular JIA patients (76.0％,19/25cases) (P ＜ 0.05),while significant improvement was not observed in oligoarticular JIA and systemic JIA patients(P ＞ 0.05).(3) Complications of upper respiratory tract infection (23.0％,12 cases) and local reaction in injection site (7.6％,4 cases) were noticed.Higher risks of tuberculosis infection and malignancy were not observed.Conclusions (1) TNF-α inhibitors treatment showed a better short-term and long-term outcome in polyarticular and oligoarticular JIA patients,and it may also improve short-term outcome in systemic JIA but with poorer long-term outcome.(2)Two evaluated systems (ACR and JADAS) were well relative,but ACR was capable to compare clinical course between different types of JIA.(3) TNF-α inhibitors treatment was relatively safe with unremarkable adverse reactions.