Background and purpose:Members of the serine protease inhibitor(SERPIN)family can influence tumorigenesis,progression,and prognosis by modulating processes such as apoptosis,invasion,metastasis,and angiogenesis in tumor cells.However,their role in pancreatic cancer remains unclear.This study aimed to investigate the impact of high expression of serine protease inhibitor A3(SERPINA3)on the proliferation,apoptosis,migration,and chemoresistance of pancreatic cancer cells and its mechanism.Methods:This study analyzed the SERPINA3 expression levels in the normal pancreatic ductal epithelial cell line hTERT-HPNE and pancreatic cancer cell lines SW1990,Capan-1,PANC-1,and ASPC-1 by real-time reverse transcription quantitative polymerase chain reaction(qRT-PCR).We established gemcitabine-resistant pancreatic cancer cell lines PANC-1/R and ASPC-1/R,and used qRT-PCR assay and cell counting kit-8(CCK-8)to determine the SERPINA3 expression levels in the constructed resistant cell lines and their parental sensitive cell lines,as well as the differences in their chemosensitivity to gemcitabine.We constructed the SERPINA3-knockdown cell line si-SERPINA with siRNA,and the negative control group si-SERPINA#NC with siRNA negative control.We used MDA assay,CCK-8 assay,EdU cell proliferation assay,transwell migration assay,matrigel invasion assay,scratch assay,and apoptotic assay to respectively detect the lipid oxidation levels,proliferation,migration,invasion,wound-healing ability,and the influence on apoptosis of the gemcitabine-resistant pancreatic cancer cells in the si-SERPINA group and the si-SERPINA#NC group.Results:Compared with normal pancreatic ductal epithelial cells hTERT-HPNE,the expression level of SERPINA3 in various pancreatic cancer cell lines was significantly increased(P＜0.05).mRNA and protein expression levels of SERPINA3 in PANC-1/R and ASPC-1/R were significantly increased compared with those in parent cells(P＜0.001).When SERPINA3 was knocked down in PANC-1/R and ASPC-1/R cells,the survival rate of the cells under different concentrations of gemcitabine chemotherapy decreased,and MDA detected that the lipid oxidation level was increased(P＜0.001).In addition,the proliferation rate of PANC-1/R and ASPC-1/R cell lines with SERPINA3 knockout,the number of migrating/invading cells and the healing rate of scratch test were significantly decreased(P＜0.01),and flow cytometry demonstrated that the number of apoptotic cells was increased(P＜0.05).These results suggest that SERPINA3 knockdown can inhibit the proliferation,migration,invasion and wound healing ability of gemcitabine-resistant pancreatic cancer cells,and promote the apoptosis of these resistant cells.Conclusion:SERPINA3 overexpression was found in various pancreatic cancer cells.SERPINA3 overexpression promoted malignant progression and chemotherapy resistance of pancreatic cancer,and interference with SERPINA3 expression promoted ferroptosis and enhanced chemotherapy sensitivity of gemcitabine-resistant pancreatic cancer cells.

Background and purpose:Members of the serine protease inhibitor(SERPIN)family can influence tumorigenesis,progression,and prognosis by modulating processes such as apoptosis,invasion,metastasis,and angiogenesis in tumor cells.However,their role in pancreatic cancer remains unclear.This study aimed to investigate the impact of high expression of serine protease inhibitor A3(SERPINA3)on the proliferation,apoptosis,migration,and chemoresistance of pancreatic cancer cells and its mechanism.Methods:This study analyzed the SERPINA3 expression levels in the normal pancreatic ductal epithelial cell line hTERT-HPNE and pancreatic cancer cell lines SW1990,Capan-1,PANC-1,and ASPC-1 by real-time reverse transcription quantitative polymerase chain reaction(qRT-PCR).We established gemcitabine-resistant pancreatic cancer cell lines PANC-1/R and ASPC-1/R,and used qRT-PCR assay and cell counting kit-8(CCK-8)to determine the SERPINA3 expression levels in the constructed resistant cell lines and their parental sensitive cell lines,as well as the differences in their chemosensitivity to gemcitabine.We constructed the SERPINA3-knockdown cell line si-SERPINA with siRNA,and the negative control group si-SERPINA#NC with siRNA negative control.We used MDA assay,CCK-8 assay,EdU cell proliferation assay,transwell migration assay,matrigel invasion assay,scratch assay,and apoptotic assay to respectively detect the lipid oxidation levels,proliferation,migration,invasion,wound-healing ability,and the influence on apoptosis of the gemcitabine-resistant pancreatic cancer cells in the si-SERPINA group and the si-SERPINA#NC group.Results:Compared with normal pancreatic ductal epithelial cells hTERT-HPNE,the expression level of SERPINA3 in various pancreatic cancer cell lines was significantly increased(P＜0.05).mRNA and protein expression levels of SERPINA3 in PANC-1/R and ASPC-1/R were significantly increased compared with those in parent cells(P＜0.001).When SERPINA3 was knocked down in PANC-1/R and ASPC-1/R cells,the survival rate of the cells under different concentrations of gemcitabine chemotherapy decreased,and MDA detected that the lipid oxidation level was increased(P＜0.001).In addition,the proliferation rate of PANC-1/R and ASPC-1/R cell lines with SERPINA3 knockout,the number of migrating/invading cells and the healing rate of scratch test were significantly decreased(P＜0.01),and flow cytometry demonstrated that the number of apoptotic cells was increased(P＜0.05).These results suggest that SERPINA3 knockdown can inhibit the proliferation,migration,invasion and wound healing ability of gemcitabine-resistant pancreatic cancer cells,and promote the apoptosis of these resistant cells.Conclusion:SERPINA3 overexpression was found in various pancreatic cancer cells.SERPINA3 overexpression promoted malignant progression and chemotherapy resistance of pancreatic cancer,and interference with SERPINA3 expression promoted ferroptosis and enhanced chemotherapy sensitivity of gemcitabine-resistant pancreatic cancer cells.

Objective To investigate the clinical efficacy of rib's strapping-traction technology using absorbable implants combined with memory-alloy ribs embracing fixator in treating multiple rib fractures under video-assisted thoracoscope.Methods A retrospective case control study was performed to analyze 178 patients with multiple rib fractures treated from January 2015 to June 2017.According to the operation method,all patients were divided into observation group (91 patients) and control group (87 patients).The observation group including 59 males and 32 females aged (39.7 ± 7.8)years adopted internal fixation by rib's strapping-traction technology using absorbable implants combined with memory alloy ribs embracing fixator under video-assisted thoracoscopic surgery.The control group including 56 males and 31 females aged (40.2 ± 6.9) years adopted internal fixation by conventional rib's clamping-traction technology using towel forcep combined with memory-alloy ribs embracing fixator under video-assisted thoracoscopic surgery.The incision length,single rib internal fixation time,number of internal fixation rib fractures,visual analogue scale (VAS) score on postoperative 1 d,postoperative chest drainage,postoperative hospital stay,and postoperative fracture healing were compared between the two groups.Results All patients were followed up for 3-30 months (mean,16.7 months).All operations were successful,with no case of thoracotomy.The observation group had shorter incision length [(4.3 ± 1.2) cm vs.(6.2 ± 1.7) cm] and single rib internal fixation time [(10.3±2.9)min vs.(14.1 ±2.3)min] than the control group (P＜0.05).There were no significant differences (P ＞ 0.05) between the two groups in number of internal fixation rib fractures [(5.7±.3.6) vs.(5.9±3.3)],postoperative chest drainage [(668.3 ±131.4)ml vs.(703.7±116.2)ml],postoperative hospital stay [(6.4 ± 1.8) d vs.(6.8 ± 1.7) d],the VAS score on postoperative 1 d [0-3 point,62％ vs.61％;4-6 point,38％ vs.39％],postoperative osteophyte formation rate [postoperative 1 month,97％ vs.95％;postoperative 3 months,100％ vs.100％].Stable chest walls and thoracic deformity recovery were seen in all patients,with no significant complications occurred.Conclusion Rib's strapping-traction technology using absorbable implants combined with memory-alloy ribs embracing fixator under video-assisted thoracoscope is convenient and has exact efficacy for multiple rib fractures,with advantage of shorter incision length and operation time over conventional thoracoscopic surgery,and thus is worthy of clinical application.

Objective To prepare and identify monoclonal antibody (McAb) against recombinant human tissue factor pathway inhibitor (rhTFPI) and to use it for measurement of TFPI by ELISA, and to evaluate the effects of the McAb on dilute prothrombin time (PT) and activated partial thromboplastin time (APTT).Methods After intrasplenic immunization of Balb/c mouse with TFPI, hybridoma technique was used to raise monoclonal antibody against rhTFPI. The McAb was well-characterized and labelled with horseradish peroxidase (HRP) by using assay of TFPI in ELISA. Furthermore, the McAb was added to normal and factor Ⅸ deficient plasma for observation of dilute PT and APTT.Results Two hybridomas (4F4, 4F8) secreting McAb against TFPI were established. The Ig class and subclass of the McAb purified from 4F8 was IgG1. Immunoblotting results indicated that the McAb4F8 only recognized a single band of TFPI with molecular weight of 34.8 KD. The results of Sandwich enzyme-linked immunosorbent assay (ELISA) by using the HRP labelled McAb4F8 showed that the mean of TFPI in normal human plasma is 103.2±11.5 μg/L. The McAb 4F8 was also proved to shorten markedly dilute prothrombin time of factor Ⅸ deficient plasma and normal plasma.Conclusions We established two hybridomas cell lines (4F4, 4F8) and obtained the McAb4F8 against TFPI and reported the levels of TFPI in healthy adult human plasma by Sandwich ELISA with HRP labelled McAb4F8 in Chinese.

Objective:To evaluate the influence of delayed application of mixed self-etching bonding agent on the tensile bond strength.Methods:Two "two-bottle,one-step" self-etching adhesives,Adper Prompt and Xeno III,were evaluated.The occlusal enamel of human third molar was cut out and the exposed dentin surface was divided in two halves by a 1 mm deep groove in the labial-lingual orientation to allow for evaluation of immediate application and delayed application(0.5,1,2,3,4,5 and 6 h) of mixed self-etching bonding agent in the same specimen,followed by composite resin built-up on the adhesive.The bonded specimens were sectioned into beams of approximately 0.9 mm2 and subsequently subjected to micro-tensile bond strength testing at a crosshead speed of 1.0 mm/min.Results:The specimens bonded with the Adper Prompt within 5 h after mixing showed no different and that with the Xeno III within 3 h after mixing showed no different.Conclusion:The effective period for "two-bottle,one-step" self-etching adhesives after mixing is limited.Different bonding agents have different effective periods.

AIM:To study the effects of lipopolysaccharide(LPS), interleukin-6(IL-6)and tumor necrosis factor ? (TNF?) on tissue factor(TF) expression of astrocytes. METHODS:Astrocytes were identified with anti-glial fibrillary acidic protein antibody. The TF activity of cell lysate was measured with one stage clotting assay. RESULTS:TF activity of astrocytes of LPS,IL-6,TNF? groups were obviously higher than that of the control group( P