Objective:To investigate the effects and possible mechanisms of caffeic acid phenethyl ester (CAPE) on the replication, amplification, and fibre formation of prions (PrP Sc). Methods:The CCK8 assay was used to detect the cell viability of the prion-infected cell model SMB-S15 after CAPE treatment for 3 days and 7 days and the maximum safe concentration of CAPE for SMB-S15 was obtained. The cells were treated with a concentration within a safe range, and the content of PrP Sc in the cells before and after CAPE treatment was analyzed by western blot. Protein misfolding cycle amplification (PMCA) and western blot were used to assess changes in PrP Sc level in amplification products following CAPE treatment. Real-time-quaking induced conversion assay (RT-QuIC) technology was employed to explore the changes in fibril formation before and after CAPE treatment. The binding affinity between CAPE and murine recombinant full-length prion protein was determined using a molecular interaction assay. Results:CCK8 cell viability assay results demonstrated that treatment with 1 μmol/L CAPE for 3 and 7 days did not exhibit statistically significant differences in cell viability compared to the control group (all P<0.05). However, when the concentration of CAPE exceeded 1 μmol/L, a significant reduction in cell viability was observed in cells treated with CAPE for 3 and 7 days, compared to the control group (all P<0.05). Thus, 1 μmol/L was determined as the maximum safe concentration of CAPE treatment for SMB-S15 cells. The western blot results revealed that treatment with CAPE for both 3 and 7 days led to a detectable reduction in the levels of PrP Sc in SMB-S15 cells (all P<0.05). The products of PMCA experiments were assessed using western blot. The findings revealed a significant decrease in the levels of PrP Sc (relative grey value) in the PMCA amplification products of adapted-strains SMB-S15, 139A, and ME7 following treatment with CAPE, as compared to the control group (all P<0.05). The RT-QuIC experimental results demonstrated a reduction in fibril formation (as indicated by ThT peak values) in CAPE-treated mouse-adapted strains 139A, ME7, and SMB-S15, as well as in SMB-S15 cells infected with prions. Furthermore, CAPE exhibited varying degrees of inhibition towards different seed fibrils formation, with statistically significant differences observed (all P<0.05). Notably, CAPE exhibited a more pronounced inhibitory effect on ME7 seed fibrils. Molecular interaction analyses demonstrated significant binding between CAPE and murine recombinant prion protein, and the association constant was (2.92±0.41)×10 -6 mol/L. Conclusions:CAPE inhibits PrP Sc replication, amplification, and fibril formation in vitro possibly due to specific interactions with the prion protein at the molecular level.

Objective:To investigate the effects and possible mechanisms of caffeic acid phenethyl ester (CAPE) on the replication, amplification, and fibre formation of prions (PrP Sc). Methods:The CCK8 assay was used to detect the cell viability of the prion-infected cell model SMB-S15 after CAPE treatment for 3 days and 7 days and the maximum safe concentration of CAPE for SMB-S15 was obtained. The cells were treated with a concentration within a safe range, and the content of PrP Sc in the cells before and after CAPE treatment was analyzed by western blot. Protein misfolding cycle amplification (PMCA) and western blot were used to assess changes in PrP Sc level in amplification products following CAPE treatment. Real-time-quaking induced conversion assay (RT-QuIC) technology was employed to explore the changes in fibril formation before and after CAPE treatment. The binding affinity between CAPE and murine recombinant full-length prion protein was determined using a molecular interaction assay. Results:CCK8 cell viability assay results demonstrated that treatment with 1 μmol/L CAPE for 3 and 7 days did not exhibit statistically significant differences in cell viability compared to the control group (all P<0.05). However, when the concentration of CAPE exceeded 1 μmol/L, a significant reduction in cell viability was observed in cells treated with CAPE for 3 and 7 days, compared to the control group (all P<0.05). Thus, 1 μmol/L was determined as the maximum safe concentration of CAPE treatment for SMB-S15 cells. The western blot results revealed that treatment with CAPE for both 3 and 7 days led to a detectable reduction in the levels of PrP Sc in SMB-S15 cells (all P<0.05). The products of PMCA experiments were assessed using western blot. The findings revealed a significant decrease in the levels of PrP Sc (relative grey value) in the PMCA amplification products of adapted-strains SMB-S15, 139A, and ME7 following treatment with CAPE, as compared to the control group (all P<0.05). The RT-QuIC experimental results demonstrated a reduction in fibril formation (as indicated by ThT peak values) in CAPE-treated mouse-adapted strains 139A, ME7, and SMB-S15, as well as in SMB-S15 cells infected with prions. Furthermore, CAPE exhibited varying degrees of inhibition towards different seed fibrils formation, with statistically significant differences observed (all P<0.05). Notably, CAPE exhibited a more pronounced inhibitory effect on ME7 seed fibrils. Molecular interaction analyses demonstrated significant binding between CAPE and murine recombinant prion protein, and the association constant was (2.92±0.41)×10 -6 mol/L. Conclusions:CAPE inhibits PrP Sc replication, amplification, and fibril formation in vitro possibly due to specific interactions with the prion protein at the molecular level.

Objective:To evaluate the effect of dexmedetomidine on the thioredoxin-interacting protein (TXNIP)/apoptosis signal-regulated kinase 1 (ASK1) signaling pathway in a mouse model of intestinal ischemia-reperfusion (I/R).Methods:Thirty-two SPF healthy adult male C57BL/6J mice, aged 8-10 weeks, weighing 18-22 g, were divided into 4 groups ( n=8 each) using a random number table method: sham operation group (Sham group), intestinal I/R group (I/R group), TXNIP inhibitor resveratrol group (Res group) and dexmedetomidine group (Dex group). The mouse model of intestinal I/R injury was developed by clamping the superior mesenteric artery for 45 min followed by 120-min reperfusion in anesthetized animals. Resveratrol 30 mg/kg was intraperitoneally injected before developing the model in Res group, and dexmedetomidine 25 μg/kg was intraperitoneally injected at 30 min before ischemia in Dex group. Blood samples were collected by cardiac puncture at the end of 120-min reperfusion, then the mice were sacrificed, and the small intestine tissues were removed for microscopic examination and for determination of the serum diamine oxidase (DAO) concentration (by enzyme-linked immunosorbent assay) and expression of TXNIP, ASK1 and cleaved-caspase-3 in small intestinal tissues (by Western blot). The apoptosis rate of intestinal epithelial cells was calculated. The intestinal damage was assessed and scored according to Chiu. Results:Compared with group Sham, the Chiu′s score, serum DAO concentrations and apoptosis rate of intestinal epithelial cells were significantly increased, and the expression of TXNIP, ASK-1 and cleaved-caspase-3 was up-regulated in group I/R ( P<0.05). Compared with group I/R, the Chiu′s score, serum DAO concentration and apoptosis rate of intestinal epithelial cells were significantly decreased, and the expression of TXNIP, ASK-1 and cleaved-caspase-3 was down-regulated in group Res ( P<0.05). Compared with I/R group, the Chiu′s score, serum DAO concentration and apoptosis rate of intestinal epithelial cells were significantly decreased, and the expression of TXNIP, ASK-1 and cleaved-caspase-3 was down-regulated in Dex group ( P<0.05). Conclusions:The mechanism by which dexmedetomidine alleviates intestinal I/R injury may be related to inhibition of the TXNIP/ASK1 signaling pathway and reduction of cell apoptosis in mice.

OBJECTIVE：To stu dy the improvement effects o f Trillium tschonos kii total sapo nins（TTM）on adjuvant-induced arthritis model rats and its mechanism. METHODS ：SD rats were randomly divided into model group （n＝44）and blank group （n＝6）. Model group was given Complete Freund ’s adjuvant to induce the adjuvant-induced arthritis model ；blank group was given constant volume of normal saline with same method. Model group were divided into model group ，Tripterygium wilfordii polyglycoside tablets group （40 mg/kg），TTM low-dose ，medium-dose and high-dose groups （50，100，200 mg/kg），with 6 rats in each group. Administration groups were given relevant medicine intragastrically ；blank group and model group were given normal saline intragastrically ，once a day ，for consecutive 9 days. After last intragastric administration ，the body weight and foot swelling degree （bilateral）were detected ，and arthritis score was performed. The contents of tumor necrosis fator α（TNF-α）， interleukin 1β（IL-1β）and IL- 6 were detected ；the pathomorphological changes of rat ankle were observed ；protein expression of NOD-like receptor protein 3 （NLRP3），apoptosis associated spot-like protein （ASC），caspase-1 were detected ；the protein expression of NLRP 3 in synovial tissue of ankle joint were also determined. RESULTS ：Compared with blank group ，foot swelling degree，serum contents of TNF-α，IL-1β and IL-6，the protein expression of NLRP 3，ASC and caspase- 1 in knee tissue as well as the protein expression of NLRP 3 in synovial tissue of ankle joint were increased significantly in model group （P＜0.05 or P＜ 0.01），while the body weight was decreased significantly （P＜0.05）. The obvious proliferation of synovial cells ，the congestion and inflammatory cell infiltration of synovial tissue were all observed. Compared with model group ，most of the above indexes were all reversed significantly in TTM groups （P＜0.05 or P＜0.01）；the pathological changes such as synovial cell proliferation ， congestion of synovial tissue and chondrocyte destruction were all relieved ，and inflammatory cell infiltration was alleviated.CONCLUSIONS：TTM can improve rheumatoid arthritis of rats， the mechanism of which may be associated with 用。E-mail：yulingling@ctgu.edu.cn inhibiting the activity of NLRP 3/caspase-1 signaling pathway and decreasing the expression of inflammatory factors TNF-α，IL-1β，IL-6.

Animals ; Cdc20 Proteins/metabolism* ; Cell Line ; Embryo, Mammalian/embryology* ; Embryonic Development ; Female ; Infertility, Female/metabolism* ; Mice ; Mutation ; Oocytes/metabolism*

Objective To explore the short-term and long-term curative effects of proprioception training for pre-term infants with functional dysphagia. Methods Seventy premature infants with functional dysphagia were ran-domly divided into a control group and an observation group, each of 35. Thirty infants of the control group and 32 from the observation group completed the whole study. Both groups were given routine medication and interventions such as touching and passive exercise training. The observation group additionally received a comprehensive interven-tion based on proprioception training. The intervention lasted from the initial stabilization of their condition to the age of 3 months. They were trained twice a day, about 20 minutes each time. The clinical manifestations and complica-tions of dysphagia were evaluated on the 28th day after their birth. At the age of 3 months, the average body mass and development quotient (DQ) were compared between the two groups. Results At the age of twenty-eight days there were significant differences between the two groups in the average number of cases of oral milk residue and coughing or oral-nasal reflux after feeding, as well as in the total number of infants with complications. However, no significant differences were found in the incidence of aspiration pneumonia or dyspnea after feeding. The average body mass and developmental quotient of the observation group were significantly better than those of the control group at the age of 3 months. Conclusion The comprehensive intervention based on proprioception training has a good clinical effect on functional dysphagia among premature infants. It can improve their life quality in both the near and longer term, and it is worth popularizing.

Objective To explore the effect of sequential and comprehensive preventative measures on the development of premature infants' intelligence.Methods A cohort of 120 premature infants was randomly divided into an observation group and a control group,each of 60.Both groups were given routine premature infant care,but the observation group was additionally provided with sequential and comprehensive preventive intervention.It included neonatal screening,inpatient-outpatient link-up,and their parents' watching CDs explaining early childhood education and health education.All of the infants were followed up from birth to 3 years old.Their adaptive capacity,fine motor skills,language acquisition,gross motor skills and social communication were evaluated at 12,24 and 36 months old using a child intelligence developmental scale for neurological development.Development intelligence quotients (DQs) were calculated and compared.Results After 12 months,significant inter-group differences were observed in adaptability and fine motor control.At 24 and 36 months old there were also significant differences in language skills.At one,two and 3 years old the average DQ of the observation group was significantly higher than that of the control group.Significant within-group differences in average DQ were observed in both groups between 1 and 2 years old,but not between 2 and 3.Conclusion Intervention within two years after birth is critical for premature infants.Timely,sequential,integrated,preventive intervention can promote the development of intelligence and better life quality for premature infants.

Objective To observe the proliferation and apoptosis of oxaliplatin-resistant colon cancer cell lines and the expression of estrogen receptor related receptor(ERR)α when tetrasoanin was down-regulated. Methods Western blot and quantitative real-time polymerase chain reaction(qRT-PCR)were used to detect the expression of tetrasoanin and ERRα of colon cancer cells and oxaliplatin resistant cells in mRNA and protein levels. ERRα inhibitor XCT790 was used to down-regulate ERRα expression, The expression of ERRα was down regulated by ERR α inhibitor XCT790,and the level of tetrasoanin,apoptosis and proliferation of L-OHP-SW620 cells were detected by Western blot, flow cytometry and MTT.Tetrasoanin expression was down regulated by siRNA, the expression, apoptosis and proliferation of L-OHP-SW620 cells AKT, p-AKT, tetrasoanin and ERRα were detected by Western blot,qRT-PCR,flow cytometry and MTT assay.Results The expression of tetrasoanin and ERRα protein in L-OHP-SW620 cell lines were higher than those in SW620 cells (t=6.127,P<0.01,t=12.579,P<0.01),The expression of tetrasoanin mRNA in L-OHP-SW620 cell line was higher than that in SW620 cell line(t=9.085,P< 0.01). The early apoptosis rate of L-OHP-SW620 cells in XCT790 group after XCT790 inhibited ERR -αexpression was higher than that in NC group(t=3.297, P< 0.01). The survival rate of XCT790 group after 72 h culture was(45.264±6.249)%,lower than that of NC group((63.364 ± 9.472)%)(t=4.537, P<0.01). Compared with NC group,p-AKT protein was up-regulated(t=8.139,P<0.01),ERRα protein was down-regulated(t=6.452,P<0.01),the apoptosis rate was(17.541±2.317)%,lower than that in the sitetrasoanin group((32.892±3.296)%)(t=4.526,P<0.01), the survival in sitetrasoanin group after 72 h culture was(49.653 ± 5.945)%, lower than that in NC group ((67.376±7.934)%)(t=3.109,P<0.05).Conclusion Tetrasoanin down-regulation and p-AKT protein up-regulation decreases ERRα protein and OHP-resistant colon cell proliferation is decreased, apoptosis is increased and drug resistance is decreased.

Objective To investigate the utility of plasma procalcitonin (PCT) as an early predictor for postoperative complications in patients who underwent elective pancreaticoduodenectomy (PD).Methods Clinical data of 87 patients who underwent elective PD in Changhai Hospital from March.1, 2016 to Dec.31, 2016 were collected.The general data, postoperative recovery, serum PCT level and white blood cell (WBC) count before, 1 d, 3 d and 5 d after PD were recorded.ROC curve was drawn and AUC was calculated to determine the cutoff value, sensitivity and specificity.Patients were divided into complication group (n=42) and noncomplication group (n=45) based on the occurrence of post-operative complications, and the comparisons between the two groups were performed.Results There were no significant differences on the age, gender, diabetes, obstructive jaundice, laboratory tests including PCT, operative time, blood loss volume during surgery and tumor type between the two groups, which were comparable.Complication group had longer hospitalization than noncomplication group (24 d vs 15 d,P<0.001), and the differences were statistically significant.In complication group, 18 patients had pancreatic fistula, 13 had peritoneal infection, 7 had gastric empty dysfunction, 8 had bleeding, 2 had bile fistula and 2 had incision infection after PD.The postoperative plasma PCT level in patients with gastric empty dysfunction, bleeding, bile fistula and incision infection was not statistically different from those in noncomplication group (all P>0.05), but the plasma PCT level in patients with pancreatic fistula and peritoneal infection on 3 d and 5 d after PD was significantly higher than those in noncomplication group, and the difference was statistically significant (all P<0.05).The combination of plasma PCT and WBC on 3 d and 5 d after PD was superior to PCT or WBC alone in predicting pancreatic fistula (sensitivity 88.9%, 72.7%;specificity 68.5%, 78.2%) and abdominal infection (sensitivity 100%, 100%;specificity 45.9%, 44.4%).Conclusions Plasma PCT could predict the occurrence of abdominal infection and pancreatic fistula after PD.The combination of PCT and WBC might be more valuable in predicting abdominal infection and pancreatic fistula.

Objective To observe the changes of proliferation and apoptosis of colon cancer cell line treated with oxaliplatin after the downregulation of ERRα and to investigate the mechanism.Methods Colon cancer cell lines Colo-205,HCT-116,SW620 and HT-29 were cultured by adherent cells and in accordance with the given intervention,they were divided into group XCT790-OHP-HCT-116(after oxaliplatin treatment,ERR αinhibitor XCT790 was administered),group siERRα-OHP-HCT-116(after oxaliplatin treatment,siERR α was transfected into HCT-116 cells and downregulated ERR αexpression),oxaliplatin intervention group(group OHP-HCT-116)and the control group(NC group)which was given no intervention.The experiment was divided into siERR αgroup with siERR α transfected with HCT-116 cells,downregulated ERR αexpression and the negative control group(siNC group)transfected with siNegative Control.Using Western blot method and real-time quantitative(qRT)-PCR for the detection of colorectal cancer cell ERRαprotein and mRNA expression,the expression of ERR αwas downregulated by ERR αinhibitors XCT790 and siERR,and apoptosis and proliferation of colon cancer cells were detected by flow cytometry and MTT.Western blot and qRT-PCR were used to detect apoptosis and proliferation-related gene proteins and mRNA expression.Results ERR αand mRNA protein in HCT-116 were higher than those of Colo-205,SW620 and HT-29 cell lines(P<0.05); in the XCT790-OHP-HCT-116 group,the early apoptosis rate was higher than those of the NC group and OHP-HCT-116 group(P<0.05),the survival rate of cell culture in 72 and 96 h in the XCT790-OHP-HCT-116 group was lower than those in the NC group and OHP-HCT-116 group(P<0.05).The siERR α HCT-116 cells transfected with down-regulation of ERR expression,siERR α -OHP-HCT-116 group early apoptosis rate was lower than those of NC group and OHP-HCT-116 group(P<0.05),siERR -OHP-HCT-116 group cells cultured for 72 and 96 h after the survival rate was lower than the NC group and OHP-HCT-116 group(P<0.05);After the downregulation of ERRαby siERR alpha transfected with HCT-116 cells,the early apoptotic rate in the group siERRα-OHP-HCT-116 was lower than that in the group NC and group OHP-HCT-116(P<0.05),the survival rate of the group siERRα-OHP-HCT-116 after 72 and 96 h were lower than those in the group NC and group OHP-HCT-116(P<0.05),siERR α was transfected into HCT-116 cells,compared with the siNC group,YAP1,p73,p63,MDM2, Capase 8,Capase 9 protein in the siERR group decreased(P<0.01),there was no significant difference in the level of mRNA(P>0.05).Conclusion The downregulation the expression of ERRαcan promote colon cancer cell apoptosis,inhibit proliferation,and enhance the killing effect of oxaliplatin on colon cancer cells.