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Objective:To evaluate the effect of hydrogen on activation of A1 astrocytes in the hippocampus of mice with sepsis-associated encephalopathy (SAE).Methods:A total of 164 clean-grade healthy male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=41 each) using a random number table method: sham operation group (group Sham), sham operation plus hydrogen group (group Sham+ H 2), group SAE and SAE plus hydrogen group (group SAE+ H 2). The SAE model was established by cecal ligation and perforation.Group Sham+ H 2 and group SAE+ H 2 inhaled 2% hydrogen starting from 1 and 6 h after operation, respectively.Twenty mice in each group were selected to observe the 7-day survival rate after operation.The remaining mice were sacrificed at 12 h after operation, and brain tissues were removed for examination of the pathological changes in hippocampal CA1 region (with a light microscope) and for determination of the apoptosis in neurons (by TUNEL), co-expression of hippocampal glial fibrillary acidic protein (GFAP) and complement C3 (by immunofluorescence staining), expression of A1 astrocyte marker C3 (by Western blot), and contents of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and high-mobility group box 1 protein (HMGB1) (by enzyme-linked immunosorbent assay). The abnormal cell ratio and apoptosis rate were calculated.Six mice in each group were selected at 7 days after operation to perform Y-Maze paradigm. Results:Compared with group Sham, the 7-day survival rate after operation was significantly decreased, the abnormal cell ratio and apoptosis rate of hippocampal neurons were increased, the contents of TNF-α, IL-6 and HMGB1 were increased, the expression of C3 was up-regulated, the number of cells coexpressing GFAP and C3 was increased, the exploration time spent in the novel arm in Y-Maze paradigm was shortened, and the preference index was decreased in group SAE ( P<0.05). Compared with group SAE, the 7-day survival rate after operation was significantly increased, the abnormal cell ratio and apoptosis rate of hippocampal neurons were decreased, the contents of TNF-α, IL-6 and HMGB1 were decreased, the expression of C3 was down-regulated, the number of cells coexpressing GFAP and C3 was decreased, the exploration time spent in the novel arm in Y-Maze paradigm was prolonged, and the preference index was increased in group SAE+ H 2 ( P<0.05). There was no significant difference in each parameter mentioned above between Sham group and Sham+ H 2 group ( P>0.05). Conclusion:The mechanism by which hydrogen improves SAE may be related to inhibiting activation of A1 type astrocytes in mice.
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Objective:To develop an evaluation index system for dynamic adjustment effect of medical service prices in public hospitals, as a set of quantitative evaluation tools for management departments to keep track of the trend in time, implement dynamic monitoring and guide decision-making.Methods:Based on the evaluation system of price adjustment effect, through the importance assessment of expert consultation and multiple index percentile method, the scoring criteria were formulated and the empirical analysis was carried out.Results:The total scores of hospital A and hospital B were 71.31 and 77.94 respectively, classified as " average" . The evaluation could basically reflect the effect of dynamic adjustment of medical service price in public hospitals.Conclusions:The evaluation has the functions of displaying differences, witnessing achievements and tracing causes. It can be used to evaluate the effect of dynamic adjustment of regional prices, to assist the regulators to keep track of trends, monitor dynamically and guide decision-making in time, and be used by hospitals in self-evaluation to find problems, improve their own operation and promote the healthy development of hospitals.
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Objective:To evaluate the effect of hydrogen-rich saline on the activity of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammomes during renal ischemia-reperfusion (I/R) in mice.Methods:Thirty male C57BL/6J mice, weighing 20-25 g, aged 6-8 weeks, were divided into 5 groups ( n=6 each) using a random number table method: sham operation group (S group), I/R group, I/R plus NLRP3 inhibitor MCC950 group (I/R+ M group), I/R plus hydrogen-rich saline group (I/R+ H group), and I/R plus MCC950 plus hydrogen-rich saline group (I/R+ M+ H group). The model of renal I/R injury was established by clipping the bilateral renal pedicles for 30 min followed by reperfusion in anesthetized animals.MCC950 20 mg/kg was intraperitoneally injected for 14 consecutive days before surgery in I/R+ M and I/R+ M+ H groups.Hydrogen-rich saline 5 ml/kg was intraperitoneally injected at 1 h after surgery in I/R+ H and I/R+ M+ H groups.The equal volume of normal sline was given instead in the other groups.Blood samples from hearts were collected at 24 h of reperfusion for determination of concentrations of blood urea nitrogen (BUN), creatinine (Cr) and kidney injury molecule-1 (Kim-1), tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 in serum (by enzyme-linked immunosorbent assay). The animals were then sacrificed, and kidney tissues were obtained for microscopic examination of pathological changes of renal tissues (with light microscopes) and for determination of cell apoptosis and expression of NLRP3, apoptosis-associated speck-like protein containing C-terminal caspase recruitment domain (ASC) and caspase-1 protein and mRNA (by Western blot or real-time polymerase chain reaction). Results:Compared with group S, the serum concentrations of Cr, BUN, Kim-1, TNF-α, IL-1β and IL-6 were significantly increased, the kidney injury score and the number of apoptotic cells were increased, and the expression of NLRP3, ASC, caspase-1 protein and mRNA was up-regulated in I/R, I/R+ M, I/R+ H and I/R+ M+ H groups ( P<0.05). Compared with group I/R, the serum concentrations of Cr, BUN, Kim-1, TNF-α, IL-1β and IL-6 were significantly decreased, the kidney injury score and the number of apoptotic cells were decreased, and the expression of NLRP3, ASC, caspase-1 protein and mRNA was down-regulated in I/R+ M, I/R+ H and I/R+ M+ H groups ( P<0.05). Compared with group I/R+ H, the serum concentrations of Cr, BUN, Kim-1 and TNF-α, IL-1β and IL-6 were significantly decreased, the kidney injury score and the number of apoptotic cells were decreased, and the expression of NLRP3, ASC, caspase-1 protein and mRNA was down-regulated in group I/R+ M+ H ( P<0.05). Conclusion:The mechanism by which hydrogen-rich saline reduces renal I/R injury is related to inhibiting the activation of NLRP3 inflammomes in mice.
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Objective@#To construct an index system for evaluating the effectiveness of dynamic pricing adjustment of medical services, for the purpose of providing a set of evaluation tools for price regulatory authorities to evaluate the effectiveness of pricing adjustment of medical services, to keep track of pricing trends, to implement dynamic monitoring and to guide decision-making.@*Methods@#Oriented to public hospitals in Guangdong province, literature analysis and Delphi method were used to construct the index system for evaluating the effectiveness of dynamic adjustment of medical service price. Descriptive analysis, consistency test and index importance evaluation were applied to statistical analysis.@*Results@#Thirty-two experts evaluated the importance of 41 alternative indicators. The index system for evaluating the effectiveness of dynamic adjustment of medical service price was finally constructed, including six structural indicators, six process indicators and six result indicators.@*Conclusions@#Experts are representative, authoritative and well-coordinated. The consultation results are reliable. The evaluation index system has high reliability and validity, and can be used to objectively evaluate the dynamic adjustment effect of medical service price.
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Objective To construct an index system for evaluating the effectiveness of dynamic pricing adjustment of medical services,for the purpose of providing a set of evaluation tools for price regulatory authorities to evaluate the effectiveness of pricing adjustment of medical services,to keep track of pricing trends,to implement dynamic monitoring and to guide decision-making.Methods Oriented to public hospitals in Guangdong province,literature analysis and Delphi method were used to construct the index system for evaluating the effectiveness of dynamic adjustment of medical service price.Descriptive analysis,consistency test and index importance evaluation were applied to statistical analysis.Results Thirty-two experts evaluated the importance of 41 alternative indicators.The index system for evaluating the effectiveness of dynamic adjustment of medical service price was finally constructed,including six structural indicators,six process indicators and six result indicators.Conclusions Experts are representative,authoritative and well-coordinated.The consultation results are reliable.The evaluation index system has high reliability and validity,and can be used to objectively evaluate the dynamic adjustment effect of medical service price.
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OBJECTIVE To investigate the cytotoxic activity of Arca subcrenata Lischke anticancer protein(ASAP)constituents on human myeloid leukemia K562 cells in vitro and analyze its anticancer mechanisms. METHODS ASAP was extracted by low temperature water and ammonium sulfate precipitation. Protein concentration of ASAP was detected by Bradford method. Morphological changes of cultured K562 cells treated with ASAP were observed under the inverted phase-contrast micro?scope. The cell and nucleus changes were analyzed by Giemsa staining. The cytotoxicity of ASAP on K562 cells was detected by MTT assay. Flow cytometry was used to detect apoptosis and cell cycle of K562 cells treated with ASAP. The expression of apoptosis and cell cycle related proteins procaspase 3, caspase 3,P53 and programmed cell death 4(PDCD4)were analyzed by Western blotting. RESULTS ASAP exhibited significant cytotoxic effect on K562 cells in a time- and concentration-dependent manner. The concentration-effect correlation coefficient of ASAP 50,100 and 200 mg · L-1 on K562 cells for 24, 48 and 72 h was 0.851,0.8977 and 0.8997,respectively. Under an optical microscope,K562 cells showed cytomorphosis,or nuclear fragmentation after treatment with ASAP 200 mg · L-1 for 48 h. Flow cytometry analysis and Giemsa staining assay indicated that apoptotic cells increased and G2/M phase cells accumulated significantly with the increase of ASAP concentration. After treatment with ASAP 200 mg · L-1 for 48 h,the early and late apoptosis cell rate increased to(32.8 ± 0.1)%and(31.2 ± 2.2)%vs control group(3.7 ± 1.1)% and (9.9 ± 0.8)%(P<0.01),respectively,and the G2/M phase cells increased to (55.2 ± 1.7)% vs (15.3 ± 0.8)% in control group(P<0.01). After treatment with ASAP 200 mg · L-1 for 0-40 h,the expression of apoptotic protein procaspase 3 was down-regulated and its active form caspase 3 was significantly up-regulated at 32 h,while PDCD4 and P53 protein expression was down-regulated significantly in 0-40 h. CONCLUSION Apoptosis and cell cycle arrest induced in G2/M phase may account for ASAP cytotoxic activity to K562 cells. K562 cell apoptosis induced by ASAP depends on caspase 3 signal pathway. Down-regulated expression of PDCD4 and P53 proteins may be related to K562 cell apoptosis and cell cycle arrest in G2/M phase by ASAP.