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OB JECTIVE To study the anti-inflammatory effect of couplet medicinals of Achyranthes bidentata -Eucommia ulmoides. METHODS Mouse macrophage RAW 264.7 were divided into blank group ，model group ，A. bidentata group（800 μg/mL），E. ulmoides group（800 μg/mL）and low- ，medium- and high- concentration groups of couplet medicinals of A. bidentata - E. ulmoides （400，800，1 600 μg/mL）. Excep for blank group and model group ，the other groups were added with corresponding drugs for 6 hours；then blank group was continued to add into the medium ，while model group was added into 10 μg/mL lipopolysaccharide （to induce the inflammatory model ）；other groups were added into corresponding drugs and 10 μ g/mL lipopolysaccharide. The levels of inflammatory factors [nitric oxide （NO），interleukin-1β（IL-1β），IL-6，tumor necrosis factor-α （TNF-α）] were detected ，and Jin ’s formula was used to evaluate the effects of A. bidentata -E. ulmoides . The expression of inducible nitric oxide synthase （iNOS），cyclooxygenase-2（COX-2），nuclear factor kappa-B （NF-κB）and inhibitor α of NF-κB （IκBα）as well as the phosphorylation of NF-κB p65，IκB kinase（IKK），p38 mitogen-activated protein kinase （p38 MAPK）， extracellular signal-regulated kinase （ERK）and c-Jun N-terminal kinase （JNK）were determined. RESULTS Compared with blank group，the level of inflammatory factors ，protein expression of iNOS and COX- 2 as well as the phosphorylation of NF-κB p65， IKK，p38 MAPK，ERK and JNK were increased significantly （P＜0.01），while the protein expression of IκBα was decreased significantly（P＜0.01）. After intervention of couplet medicinals of A. bidentata -E. ulmoides ，the level of inflammatory factors ，the expression or phosphorylation of above proteins were reversed significantly （P＜0.05 or P＜0.01），and couplet medicinals of A. bidentata-E. ulmoides had a synergistic effect. CONCLUSIONS The couplet medicinals of A. bidentata -E. ulmoides have synergistic anti-inflammatory effect on RAW 264.7 cells. Its mechanism may be related to the inhibition of NF-κB/MAPK signaling pathway related protein expression.
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Objective To isolate and identify exosomes from serum samples of the patients with polymyositis / dermatomyositis (PM/ DM),and analyze their protein composition preliminarily.Methods Exosomes from serum samples of the patients with PM/DM were isolated and purified by the ExoQuickTM kit.The morphological characteristics and particle size of exosomes were determined by transmission electron microscope (TEM) and NanoSight analyzer,respectively.The surface markers of exosomes such as CD9,CD81 and Flotillin-2 were identified by western blot.The concentration and composition of exosome protein were determined by the BCA method and SDS-PAGE,respectively.Results The exosomes from serum samples of PM/DM patients displayed round or oval vesicles with membrane structure under TEM,and their diameter range was about (92 ± 67) nm.western blot showed that these exosomes expressed CD9,CD81 and Flotillin-2.The total protein concentrations of exosomes in the patients with PM/DM and healthy controls were 14.68 (6.00,32.55) μg/μL and 14.09 (8.00,23.28) μg/μL,respectively.SDS-PAGE showed that high-abundance proteins enriched in 55-70 kD in both PM/DM patients and healthy controls,and that there were different bands in 40-55 kD between them.Conclusion Exosomes are isolated from serum samples of the patients with PM/DM successfully,and their protein concentration and composition are analyzed preliminarily,which provides the experimental evidences for further finding differential proteins.
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Background and purpose:Papillary renal cell carcinomas (PRCC) is relatively infrequent, and there are few related researches in China. This study aimed to summarize the clinical and pathological features of PRCC, and evaluate prognostic factors for patients treated with surgery.Methods:A total of 64 patients who under-went surgery for PRCC were retrospectively assessed. PRCC tissue slides from each patient were reviewed for type (ⅠorⅡ), grade, TNM stage, coagulative tumor necrosis and microvascular invasion. We estimated overall survival using the Kaplan-Meier method. Multivariate analysis was done according to the Cox proportional hazards model of factors statistically signiifcant on univariate analysis. Results:The incidence rate of ENE was 6.04%in RCC, the median age was 55 (range 22 to 78) years. The comparison of the 22 (534.4%) typeⅠPRCCs and 42 (65.6%) typeⅡPRCCs revealed that typeⅡtumors were associated with a greater stage and grade more often. The median follow-up was 46 months (range 19 to 133). Of the 64 patients, 14 died, (4.5%) with typeⅠand 13 (31.0%) with typeⅡtumors (P=0.018). The overall survival rate was 85.7%in typeⅠtumors and 55.8%in typeⅡtumors, respectively. Univariate analysis identiifed symptoms at presentation, tumor type, TNM stage and grade as prognostic factors. On multivariate analysis only metastatic RCC remained associated with decreased overall survival (HR:14.78, P=0.004). Conclusion:The per-centage of PRCC is lower and typeⅡPRCC is relatively common compared with foreign data. Metastasis at diagnosis is an independent predictive parameter of overall survival in Chinese patients with PRCC.