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Objective To explore the potential effects of neutrophil extracellular traps (NETs) on rheumatoid arthritis synovial fibroblasts (RA-FLSs).Methods The synovial tissues of RA patients were isolated and cultured in vitro.Peripheral blood neutrophils were extracted from healthy volunteers and used to stimulate NETs' formation,following with NETs' extraction.MTS proliferation assay was used to evaluate the effect of NETs on the proliferation of RA-FLSs.QRT-polymerase chain reaction (q-PCR) was used to determine the expression of connective tissue growth factor (CTGF) mRNA in cells treated with NETs-stimulated RA-FLSs for 60 h.The results were processed using paired sample t-test and one-way analysis of variance (ANOVA).Results The isolated and purified neutrophils could form NETs by in vitro stimulation.The concentration of extracted NETs-DNA was 58.5 ng/μl (1×106 cells).Compared with the control group (0 μl NETs),NETs could promote the proliferation of RA-FLSs.With the increase of NETs' concentration,the proliferation of RA-FLSs was also enhanced (F=99.519,P＜0.05).Compared with the control group (0 μl NETs),10 μl NETs could significandy promote the proliferation of RA-FLSs (t=-12.226,P＜0.01).Pretreatment of NETs with DNase Ⅰ inhibited its effect on promoting the proliferation of RA-FLSs (t=-2.376,P=0.049),NETs stimulated the upre-gulation of CTGF mRNA expression in RA-FLSs [(30.7±0.5),t=12.13,P＜0.01].Conclusion NETs can promote the proliferation of RA-FLSs and stimulate the up-regulation of CTGF mRNA in RA-FLSs in vitro.
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Objective To study the function of interleukin (IL)-25 for rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) differentiation as well as on the expression of extracellular regulating protein kinase (ERK) and matrix metalloproteinases-3 (MMP-3).Methods The differences on ERK1/2 and MMP-3 protein levels were tested in RA-FLS of RA patients and healthy controls,then IL-17A (10 ng/ml) was tested when the RA-FLS were co-stimulated with different concentrations of IL-25 (0.01,0.1,1 and 10 ng/ml) and IL-17A(10 ng/ml) for 24 hours respectively.The expression of ERK1/2 and MMP-3 protein was detected by the Western blot.T test was used for the comparison between different groups.Results The expression of ERK1/2 (1.71±0.17) and MMP-3 (0.50±0.13) proteins in RA-FLS was higher than the healthy controls (0.50±0.15,0.17±0.05) (t=-9.13,P＜0.01 and t=-4.10,P＜0.05),after stimulated with IL-17A,the expression of ERK1/2 (0.77±0.22) and MMP-3 (0.59±0.13) proteins in RA-FLS were increased compared with the untreated groups (0.18±0.35,0.04±0.03) (t=-4.69,P＜0.01 and t=-7.47,P＜0.01).With increase of the concentration on IL-25,the level of ERK1/2 (0.54±0.26,0.48±0.18,0.48±0.23,0.23±0.06) and MMP-3 (0.58±0.09,0.59±0.14,0.21±0.04,0.04±0.02) in RA-FLS which were stimulated by IL-17A was decreased slowly (t=4.22,P＜0.05 and t=4.95,P＜0.01 and t=7.47,P＜0.01).Conclusion IL-25 can inhibit the stimulation of IL-17A on ERK1/2 and MMP-3 fractionally,which implies that it may take part in the development of RA through this pathway and may be a target for the RA treatment.
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Objective To investigate the possible differences in safety and efficacy between re-use and initial use in patients with ankylosing spondylitis (AS) treated with tumor necrosis factor inhibitors (TNFi). Methods From October 2016 to October 2017, 82 patients with AS who were admitted to the Second Hospital of Lanzhou University were studied. Among them, 57 patients used TNFi for the first time and 25 patients reuse it after the interruption. After 3 months of standardized use of TNF-inhibitor, we compared the efficacy indicators [visual analogue scale/score (VAS), morning stiffness, bath ankylosing spondylitis disease activity index (BASDAI), Bath ankylosing spondylitis functional index (BASFI), Bath ankylosing spondylitis metroloty index (BASMI), ankylosing spondylitis disease activity score (ASDAS), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) 0 and safety events between the two groups. T test and covariance analysis were used. Results The efficacy indexes of the two groups after treatment were significantly improved, the difference was statistically significant (P<0.05) compared with the baseline [Before and after treatment in the first treatment group: ESR: (40±31) mm/1 h, (8±8) mm/1 h, CRP: (28±35) mg/L, (5±9) mg/L, VAS: (6.5±1.6), (2.0 ±1.7), Morning stiff time: (0.6 ±0.4) h, (0.1 ±0.2) h, BASDAI: (5.0 ±1.3) h, (1.6 ±1.2) h, BASFI: (4.1 ±2.3), (1.3±1.3), BASMI: (2.6±2.0), (0.8±1.0), ASDAS: (3.5±0.8), (1.2±0.7); Before and after treatment in the re-use group: ESR: (39 ±33) mm/1 h, (9 ±10) mm/1 h, CRP: (28 ±28) mg/L, (5 ±6) mg/L, VAS: (6.6 ±1.9), (1.6 ±1.0), Morning stiff time:(0.6±0.4) h, (0.1±0.1) h, BASDAI:(5.1±0.8), (1.4±1.4), BASFI (5.1±2.2), (1.3±1.4), BASMI:(3.4 ±1.8), (1.0 ±0.9), ASDAS: (3.6 ±0.8), (1.2 ±0.4)]. But there was no statistical significant difference between the two groups in patients after treatment (P>0.05). Conclusion Patients with AS who re-uses TNFi after discontinuation could achieve the same safety and efficacy as they first use it.
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Objective To observe the impacts of Ruangan-Lidan Tang on immune function for liver cancer patients receiving TACE combined with 3D conformal radiotherapy, evaluation of short-term efficacy, survival, to explore the changes of cellular immune function in the treatment. Methods 86 cases of liver cancer patients were randomly divided into treatment group and control group were taken in 3D conformal radiotherapy to 50-60GY/5-6 week for fourth weeks after TACE, the treatment group taking Ruangan-Lidan Tang intervention, control group without the use of traditional Chinese medicine intervention.Before the start, after 1 weeks of TACE, within 3 days before radiotherapy,end of radiotherapy, a total of 4 times for T cell differentiation antigen were measured, and between two groups were compared with t test, 2 months after radiotherapy CT or MRI evaluation of curative effect, survival. Results 5 cases of loss of cases, the total cases were 81 patients, including 40 cases of treatment group, 41 cases in After 1 weeks of TACE, CD3+, CD4+, CD8+of the treatment group were (55.15±4.76)%, (31.88±5.50)%, (22.00±3.18)% of the control group were (53.39±5.00)%, (30.49±5.94)%, (23.39±3.41)%. There was no significant difference between two groups (P>0.05); Before radiotherapy, CD3+, CD4+, CD8+ of the treatment group were (59.05±5.91)%, (40.55±6.17)%, (18.63±3.14)%, of the control group were 55.88±4.65%,33.88±4.41%,22.00±3.78%. There was significant difference between two groups(P0.05). In the treatment group, the median survival time was 22 months,the average survival time was 21.7 months, the half year survival rate was (97.5± 2.5)%(39/40);In the control group, the median survival time was 20 months, the average survival time was 19.3 months, the half year survival rate was (94.9±3.5)% (37/41). Follow-up survival analysis, treatment group than in the control group increased, but there was no statistical difference (P=0.132, P>0.05). Conclusion Ruangan-Lidan Tang can improve the immune function of liver cancer patients receiving TACE combined with radiotherapy, but in the Short-term efficacy and survival did not reflect advantage.