ABSTRACT
OBJECTIVE：To ex plore the protective effects of Longbie capsule contained serum （called“LBJN”for short ）on the apoptosis of chondrocytes induced by YAP inhibitor verteporfin and its mechanism. METHODS ：Primary human knee osteoarthritis（OA）chondrocytes were extracted by two-step enzymatic digestion ，and then identif ied by toluidine blue staining and type Ⅱ collagen immunofluorescence staining. The effects of 2，5 μmol/L verteporfin alone or combined with 5％LBJN on cell apoptosis were detected by flow cytometry. Solvent control （0.1％ DMSO）and 5％ LBJN were set. Western blot assay was adopted to detect the expression of apoptosis related proteins （YAP，Bcl-2，cleaved-caspase-3） after treated with 0.1％DMSO（solvent control ），2 μmol/L verteporfin，2 μmol/L verteporfin+5％LBJN 和 0（blank control ），2.5％ LBJN and 5％ LBJN for 48 h. The expression of autophagy related proteins （mTOR，Beclin-1，LC3A/B） after treated with 0 （blank control ），2.5％，5％ LBJN for 48 h were det ected by Western blot assay. RESULTS ：The isolated cells accorded with the characteristics of chondrocytes. Compared with 0.1％DMSO， the apoptosis rates of cells were increased significantly after treated with 2，5 μmol/L verteporfin（P＜0.05），and the effects of the two concentrations were similar （P＞0.05）. Compared with verteporfin alone ，2，5 μmol/L verteporfin combined with 5％LBJN could significantly decrease the apoptotic rate of cells （P＜0.05）. Compared with 0.1％DMSO，the protein expression of YAP and Bcl-2 were decreased significantly after treated with 2 μ mol/L verteporfin （P＜0.05）， while the protein expression of cleaved-caspase-3 were increased significantly （P＜0.05）. Compared with 2 μmol/L verteporfin，protein expression of YAP and Bcl-2 were increased significantly after treated with 2 μmol/L verteporfin+5％LBJN（P＜0.05），while the protein expression of cleaved-caspase-3 were decreased significantly （P＜0.05）. Compared with blank control ，the protein expression of YAP ，Bcl-2 and Beclin-1 were increased significantly after treated with 2.5％，5％LBJN（P＜0.05），while protein expression of cleaved-caspase- 3 and mTOR were decreased significantly （P＜0.05）. CONCLUSIONS ：LBJN can block the apoptosis of chondrocytes induced by YAP inhibitor verteporfin ，and its mechanism may be related to regulating the expression of apoptosis related proteins and enhancing autophagy of chondrocytes.
ABSTRACT
BACKGROUND: Total knee arthroplasty is a procedure for treatment of knee osteoarthritisa with standardized, mature technology and affirmative efficacy. Total knee arthroplasty can result in overt excessive bleeding, decreased hemoglobin levels, patient mouth infection and other complications. As a new technology, autologous blood transfusion device can effectively reduce the rate of blood transfusion through reinfusing the unwashed and filterable drainage blood after operation. Up to now, no systematic reviews incorporating meta-analyses have found directly sufficient evidence to compare autologous blood transfusion drainage and no drainage after primary total knee arthroplasty. OBJECTIVE: To study the clinical efficacy, safety and potential advantages of the application of autologous blood transfusion device/no drainage based on the meta-analysis. METHODS:PubMed, Embase, the Cochrane Library, CBMdisc, China HowNet, VIP, Wanfang database were searched comprehensively by computer. The search strategies were developed by the way of MeSH terms combining with free words: “total knee replacement” OR “total knee arthroplasty” OR “total knee prosthesis” OR “unicompartmental” OR “unicondylar” OR “unicompartmenta” OR “arthroplasty, replacement, knee” [MeSH terms] AND “autologous blood transfusion” OR “Autotransfusion” OR “blood transfusion, autologous” [MeSH Terms] OR “Intraoperative Blood Salvage” OR “Intraoperative Blood” OR “Postoperative Blood Salvage” OR “Intraoperative Blood Cel Salvage” OR “Operative Blood Salvage” [MeSH Terms]. Data included in the final literature were analyzed using RevMan 5.3.5 software recommended by Cochrane. The main outcome measure was the rate of transfusion. The secondary outcome measures were the average change in hemoglobin, hemoglobin levels at the 3rd day, hospitalization time and intraoperative mouth infection rate. RESULTS AND CONCLUSION:Five randomized controlled trials, a total of 667 patients were enroled. Meta-analysis results showed that there were no significant differences in the transfusion rate (OR=0.73, 95%CI: 0.47-1.13;Z=1.41,P=0.16), average change in hemoglobin (WMD=0.20, 95%CI:-0.28-0.68;Z=0.82,P=0.41), the hemoglobin levels at the 3rdday (WMD=0.41, 95%CI:-0.26-1.09;Z=1.20,P=0.23), hospitalization time (OR=1.01, 95%CI: 0.06-16.27;Z= 0.01,P=1.00), intraoperative mouth infection rate (OR=1.01, 95%CI: 0.06-16.27;Z=0.01,P=1.00) between the postoperative use of autologous blood transfusion and no drainage. These results suggest that the meta-analysis of outcome measures has not provided the evidence-based medical support for the clinical efficacy of autologous blood transfusion device (including blood transfusion rate, the average change in hemoglobin, average hemoglobin change at the 3rd day, hospitalization time). Given the inherent limitations of the quality of the included studies and the publication bias, future high-quality, large-volume, multi-center randomized controled trials are awaited to confirm and update the findings of this analysis.
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OBJECTIVE A surveillance study was performed for nosocomial infections in order to investigate the change in antimicrobial resistance of Klebsiella pneumoniae,especially the strains isolated from 1999 to 2004.METHODS K-B test was used for the antibiotics susceptibility test and the results were read based on National Committee for Clinical Laboratory Standards(NCCLS) of the USA.The situation of ESBLs-producing strains of K.pneumoniae was investigated.RESULTS Totally 326 K.pneumoniae strains showed the highest susceptibility to imipenem.Ceftazidime,cefepime,and cefoperazone/sulbactam also showed excellent activity against K.pneumoniae.The prevalence of ESBLs from 326 strains was 20.2%.CONCLUSIONS It is important to study the drug resistance in nosocomial infections by K.pneumoniae.