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Objective:The effect of intermedin (IMD) on ATP-induced activation of inflammatory bodies and pyroptosis of cells and its mechanism were studied using lipopolysaccharide (LPS)-sensitized mouse macrophage line RAW 264.7.Methods:The cells were divided into the control groups, the LPS groups, LPS+IMD groups, and LPS+IMD+LY294002 groups. The expression of interleukin (IL)-1β and IL-18 and the activation of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammatory cells were detected by real-time PCR and western blotting, and the pyroptosis of cells was detected by propidium iodide (PI) staining. The measurement data was represented by MS± SD, and the inter-group difference was compared with ANOVA calculations, and P<0.05 represented the difference with statistical significance. Results:Compared with the control group [(0.83±0.09) vs (0.49±0.04)], the ratio of phosphorylated phosphatidylinositol-3-kinase, p-PI3K)/phosphatidylinositol-3-kinase (PI3K) (0.44±0.05) and p-Akt/Akt (0.27±0.06) in the LPS group was significantly decreased. The ratios of p-PI3K/PI3K (1.22±0.18) and pAkt/Akt (0.83±0.09) in LPS+IMD group was significantly increased ( F=31.40, P<0.001; F=50.88, P<0.001). Compared with the control group, the mRNA and protein expressions of IL-1β, IL-18 and NLRP3 inflammasome (NLRP3, caspase-1, ASC) in RAW264.7 cells were up-regulated in the LPS group (LPS and ATP). Compared with LPS group, IMD treatment inhibited the expression of inflammatory cytokines IL-1β, IL-18 and NLRP3 inflammasome, which was blocked by LY294002, a blocker of PI3K/Akt pathway. The results of real-time PCR showed that the relative expression of IL-1β mRNA was (1.00±0.11) in the control group, (8.32±0.61) in the LPS group, (8.32±0.55) in the LPS+IMD group, and (7.23±0.41) in the LPS+IMD+LY group ( F=15.42, P<0.001). The relative expression of IL-18 mRNA in the control group was (1.00±0.17), (1.82±0.21) in the LPS group, (1.14±0.15) in the LPS+IMD group, and (1.53±0.11) in the LPS+IMD+LY group respectively ( F=18.16, P<0.001). The relative expression of NLRP3 mRNA in the control group was (1.00±0.13), (2.58±0.18) in the LPS group, (1.07±0.17) in the LPS+IMD group, and (1.33±0.32) in the LPS+IMD+LY group respectively ( F=15.98, P< 0.001); The relative expression of caspase-1 mRNA in the control group was (1.00±0.09), (6.20±0.19) in the LPS group, (3.43±0.06) in the LPS+IMD group, and (5.50±0.45) in the LPS+IMD+LY group respectively ( F=18.39, P<0.001). The relative expression of ASC mRNA in the control group was (1.00±0.21), (4.58±0.48) in the LPS group, (2.07±0.51) in the LPS+IMD group, and (3.33±0.32) in the LPS+IMD+LY group respectively ( F=15.19, P<0.001). Western blotting results showed that the relative expression of IL-1β protein was as follows: (100%) in the control group, [(188±14)%] in the LPS group, [(112±11)%] in the LPS+IMD group, and [(171±27)%] in the LPS+IMD+LY group respectively ( F=21.25, P<0.001). The relative expression of IL-18 protein in the control group was 100%, [(183±16)%] in the LPS group, [(115±19)%] in the LPS+IMD group, and [(179±23)%] in the LPS+IMD+LY group respectively ( F=19.62, P<0.001). The relative expression of NLRP3 protein was 100% in the control group, [(149±15)%] in the LPS group, [(106±10)%] in the LPS+IMD group, and [(144±15)%] in LPS+IMD+LY group respectively ( F=14.35, P<0.001). The relative expression of ASC protein was 100% in the control group, [(188±12)%] in the LPS group, [(110±18)%] in the LPS+IMD group, and [(192±8)%] in the LPS+IMD+LY group ( F=15.79, P<0.001). Conclusion:IMD inhibits the activation of NLRP3 inflammasome and cell pyroptosis by regulating PI3K/Akt activity.
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Objective To explore the mechanism of the protection of acidic fibroblast growth factor (aFGF) for hippocampal astrocytes from injury induced by gentamicin. Methods Hippocampal astrocytes were isolated from newborn (24 hours) Sprague-Dawley rats, puri-fied, and identified with glial fibrillary acidic protein (GFAP) immunofluorescence. The third generations were cultured for 3 days and divid-ed into 3 groups:control group was cultured routinely, injury group was cultured with 2.0 g/L gentamicin for 24 hours, and protection group was cultured with 4.25μg/L aFGF for 24 hours and then cultured with 2.0 g/L gentamicin for 24 hours. Western blotting was adopted to de-tect the expressions of P38, extracellular signal-regulated kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK)1/2. Results Hippocampal as-trocytes were culturated successfully with the purity above 95%. The ERK1 increased in the injury group compared with the control group (P0.05). Conclusion The mitogen-activated protein kinase signal pathway, especially P38 and ERK1, may associate with the protection of aFGF for hippocampal astrocytes from injury induced by gentamicin.
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Objective To evaluate the level situation of clinical doctors graduating from mil-itary medical university within a certain period, understand their career development, and explore and analyze the problems and deficiencies in doctoral education and training to provide reference for deep-ening the reform of clinical disciplines doctoral education. Methods Stratified random samples of 9 Hospitals were selected and the established evaluation index system of clinical discipline quality tracking and investigation questionnaire was used to evaluate doctoral quality. Evaluation was surveyed by self rating and other rating combination. Quality evaluation system contains 4 first level indicators such as the military and political quality, clinical, teaching and research level, the development po-tential as well as 15 second-level indexes. The scores of self-assessment and other evaluation were compared by Pearson rank correlation analysis. The corresponding indicator scores of different doctorate types were compared using the test of variance and the subject research and published papers were analyzed by χ2 inspection. Results The comprehensive score of graduated doctors is excellent, with self-evaluation score (92.72±7.06) and other evaluation score (93.61±8.05). Correlation coefficient is 0.33(P=0.04); The academic doctors have done better in publishing papers(χ2=5.97, P=0.01) and undertaking subject research(χ2=6.08, P=0.00), but poorer in clinical work compared with the doctors of professional degree. The assessed groups are inadequate in publishing high level papers and in un-dertaking research projects and doctors of different degree types have different cognition of the ele-ments of improving the quality. Conclusions Cultivating the doctors' clinical ability and innovation ability are the cores of deepening the reform of clinical doctoral education. We should focus on inno-vation ability , value the cultivation of the clinical professional doctoral degree and explore various joint evaluation systems to attain the goal of enhancing the education quality of doctors.