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Objective To investigate the role and potential mechanism of high mobility group box-1 protein (HMGB1) on improving the chemosensitivity of gemcitabine-resistant pancreatic cancer PANC1 ceils.Methods Gemcitabine-resistant pancreatic cancer PANC1 (PANC1-GR) cell line was established by using increased gradient concentration of gemcitabine.The si-HMGB1-PANC1 and si-HMGB1-PANC1-GR cells were established by the transfection with HMGB1 siRNA using liposome.The 50％ inhibitory concentration (IC50) and Resistance index (RI) of gemcitabine in 4 PANC1 cell lines with or without HMGB1 siRNA transfection were determined and calculated by CCK-8 assay.Western blot assay was used to detect the protein expression of HMGB1 in PANC1 and PANC1-GR cells and the expression of autophagy marker protein Beclin1 in the 4 PANC1 cell lines.Flow cytometry assay was used to evaluate the apoptosis rate of 4 pancreatic caner cell lines.Results The gemcitabine-resistant pancreatic cancer cell line PANC1-GR was successfully established,which could grow stably and passage in media with 100 μmol/L gemcitabine.The IC50 of gemcitabine in PANC1,PANC1-GR,si-HMGB1-PANC1,and si-HMGB1-PANC1-GR cells lines were (4.7 ±0.4) μmoL/L,(166.5 ± 13.6) μmol/L,(3.2 ± 0.3) μmol/L,and (52.4 ± 8.4) μmol/L,respectively.The IC50 in PANC1-GR wassignificantly higher than that in PANC1,while the IC50 in the transfected cells was significantly lower than that in untransfected cells,and the differences were both statistically significant (bothP ＜ 0.01).The RI value of gemcitabine in transfected and untransfected PANC1-GR cells was 35.4 and 16.4.The relative protein levels of HMGB1 in PANC1 and PANC1-GR were 0.17 ± 0.08 and 0.38 ± 0.11.The expression of HMGB1 in PANC1-GR was obviously higher than that in PANC1,and the difference was statistically significant (P＜0.01).The relative protein levels of Beclin1 in PANC1,PANC1-GR,si-HMGB1-PANC1 and siHMGB1-PANC1-GR cells were 2.68 ± 0.23,3.28 ± 0.15,0.68 ± 0.23 and 0.78 ± 0.11,which in two transfected cells was greatly lower than those in untransfected cells.The apoptosis level was (34.58± 3.14)％,(79.56±3.58)％,(19.41± 1.53)％,and (34.57±2.94)％.The apoptosis level in the 2 transfected cell lines were significantly higher than those in the 2 untransfected cell lines,and the differences were both statistically significant (P ＜ 0.01).Conclusions The inhibition of HMGB1 could improve the chemosensitivity of gemcitabine in pancreatic cancer PANC1 cells,which might be mediated by the activation of autophagy.
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OBJECTIVE To investigate the reasonable use of antibacterial in department of orthopedics.METHODS The patients in our department were registered according to the inventory of antibacterial use made by the hospital infection monitoring network of Medical Administration Bureau,Ministry of Health of the People′s Republic of China,the(operative) incision was classified according to Clinical Surgery.The data of reasonable use of antibacterial were(obtained) after evaluation and(analysis).RESULTS Of the 257 patients in our study,79.37% of them were used antibacterial,20.62% were not used antibacterial,24.50% were used antibacterial for therapy,75.49% were used antibacterial for infection(prevention);95.09% were used systematically;74.50% were reasonable(usage,) 25.49% were unreasonable usage;of the 204 patients who were used antibacterial,only 34 patients were tested by drug sensitivity trial,and the others were used by experience.CONCLUSIONS In order to use antibacterial reasonably,we suggest two points,one is that the medicine management department should workout a feasible antibacterial use institution and mass(evaluation) criterion,take mass examination regurally,and connect the bonus to the examination result;the other is that we should enhance the management of antibacterial market,avoid the improper competition,rise the morality view of the doctor,and suggest them to use antibacterial reasonably(according) to the scientific theory.