BACKGROUND:Studies have confirmed that Pterocarya hupehensis skan total flavonoids(PHSTF)can improve the level of collagen-induced arthritis in rats,but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.OBJECTIVE:To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation,migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathwayMETHODS:Fibroblast-like synoviocytes were divided into control group,lipopolysaccharide group,lipopolysaccharide+low-,medium-,and high-dose PHSTF groups(10,20,and 40 μg/mL),lipopolysaccharide+Wnt pathway inhibitor DKK1 group,and lipopolysaccharide+Wnt pathway inhibitor DKK1+high-dose PHSTF group(40 μg/mL).The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes,and the final drug concentration and time were screened.Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes.Cell scratch assay,EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes.Western blot assay was used to detect the protein expression levels of Wnt3a,β-catenin,tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,Bax and Bcl-2 in fibroblast-like synoviocytes.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability decreased significantly when the concentration of PHSTF was＞40 μg/mL(P＜0.01).Therefore,the drug concentration of≤40 μg/mL was selected for subsequent experiments.(2)Compared with the lipopolysaccharide group,the wound healing rate,cell clone formation rate and the number of EDU-positive cells in the low-,medium-and high-dose PHSTF groups were significantly reduced,while the apoptosis rate was significantly increased(P＜0.05-0.01).(3)Western blot results showed that compared with the lipopolysaccharide group,low-,medium-and high-dose PHSTF significantly inhibited cellular Wnt3a,β-catenin,cellular tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,and Bcl-2 protein expression,and promoted the expression of Bax protein(P＜0.01).(4)Compared with the DKK1 group,the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a,β-catenin,matrix metalloproteinase 2,matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax(P＜0.01).To conclude,PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

BACKGROUND:Studies have confirmed that Pterocarya hupehensis skan total flavonoids(PHSTF)can improve the level of collagen-induced arthritis in rats,but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.OBJECTIVE:To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation,migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathwayMETHODS:Fibroblast-like synoviocytes were divided into control group,lipopolysaccharide group,lipopolysaccharide+low-,medium-,and high-dose PHSTF groups(10,20,and 40 μg/mL),lipopolysaccharide+Wnt pathway inhibitor DKK1 group,and lipopolysaccharide+Wnt pathway inhibitor DKK1+high-dose PHSTF group(40 μg/mL).The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes,and the final drug concentration and time were screened.Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes.Cell scratch assay,EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes.Western blot assay was used to detect the protein expression levels of Wnt3a,β-catenin,tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,Bax and Bcl-2 in fibroblast-like synoviocytes.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability decreased significantly when the concentration of PHSTF was＞40 μg/mL(P＜0.01).Therefore,the drug concentration of≤40 μg/mL was selected for subsequent experiments.(2)Compared with the lipopolysaccharide group,the wound healing rate,cell clone formation rate and the number of EDU-positive cells in the low-,medium-and high-dose PHSTF groups were significantly reduced,while the apoptosis rate was significantly increased(P＜0.05-0.01).(3)Western blot results showed that compared with the lipopolysaccharide group,low-,medium-and high-dose PHSTF significantly inhibited cellular Wnt3a,β-catenin,cellular tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,and Bcl-2 protein expression,and promoted the expression of Bax protein(P＜0.01).(4)Compared with the DKK1 group,the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a,β-catenin,matrix metalloproteinase 2,matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax(P＜0.01).To conclude,PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

Objective To investigate the impact of county-level"Unified ECG Network"construction on the treatment efficiency and clinical outcomes of patients with acute ST-segment elevation myocardial infarction(STEMI).Methods A retrospective analysis was conducted on the clinical data of STEMI patients from Beichuan County and Yanting County in Mianyang City,and Jiange County in Guangyuan City,Sichuan Province,during the 18 months before(128 cases)and 18 months after(187 cases)the establishment of the"Unified ECG Network."Differences in demographic characteristics,treatment efficiency,therapeutic methods,and clinical outcomes between the two groups were compared.Results There was no statistically significant difference in general demographic characteristics between the two groups(all P＞0.05).Compared with the pre-construction group,the post-construction group showed significantly shorter times in initial ECG completion[5(3,7)min vs.6(4,8)min],initial ECG diagnosis[3(2,4)min vs.5(2,6)min],first medical contact to preliminary diagnosis[10(9,12)min vs.13(11,15)min],network hospital door-in-door-out time[21(19,23)min vs.26(23,30)min],and first medical contact to wire-crossing time[(94.82±11.87)min vs.(107.97±18.39)min](allP＜0.001).The proportion of patients bypassing the emergency department and coronary care unit significantly increased(64.17％vs.32.81％,P＜0.001).The proportion of patients undergoing emergency percutaneous coronary intervention significantly increased(72.73％vs.51.56％,P＜0.001),while the proportions of thrombolytic therapy and non-reperfusion therapy significantly decreased(both P＜0.05).Additionally,in-hospital mortality rate,Killip class≥Ⅱ proportion,incidence of major adverse cardiovascular events,and average length of hospital stay were all significantly reduced(all P＜0.05).There were no statistically significant differences among the three county-level chest pain centers in terms of major treatment efficiency,therapeutic strategies,or clinical outcomes(all P＞0.05).Conclusions The construction of the county-level"Unified ECG Network"can significantly improve the treatment efficiency of STEMI patients,optimize reperfusion therapy strategies,improve clinical outcomes,and demonstrate substantial clinical promotion value.

Objective To explore the effect and mechanism of lentinan(LNT)on hepatic tissue ferroptosis in mice exposed to sodium arsenite(SA).Methods C57BL/6N male mice were exposed to SA low-dose,SA high-dose,and LNT intervention combined with SA high-dose,then,hematoxylin and eosin(HE)staining was applied to assess pathological liver tissue damage;Enzyme-linked immunosorbent and Western blot were used to detect the content or expression of tumor necrosis factor α(TNFα),interleukin-6(IL-6),ferritinophagy or ferroptosis biomarkers.Results Compared with the control group,SA exposure induced the elevated levels of TNFα,IL-6,ferritinophagy biomarker ferritin heavy chain 1(FTH1)and microtubule-associated protein 1 light chain 3B(MAP1LC3B)in mice liver tissue,while levels the ferroptosis biomarker GPX4 decreased(P＜0.05).Compared with SA high-dose groups,LNT intervention showed the reduced pathological liver damage and the downregulated levels of TNFα,IL-6,FTH1,and MAP1LC3B,while the level of GPX4 upregulated(P＜0.05).Western blot experiment showed that LNT intervention antagonized the upregulated levels of FTH1,and autophagy biomarker LC3B/A,and antagonized the increased co-expressions of FTH1 with LC3B or Ub protein in SA high-dose group(P＜0.05).Conclusions LNT antagonizes SA-exposed hepatic pathological injury and ferroptosis in mice,possibly associated with inhibition of TNFα-ferritinophagy signaling.

Aim To investigate the inhibitory effect of the protein kinase D(PKD)-specific inhibitor CRT0066101 on hepatocellular carcinoma(HCC)and its underlying molecular mechanisms,providing new theoretical insights and therapeutic strategies for targe-ted HCC treatment.Methods HCC cell lines were treated with varying concentrations of CRT0066101.The inhibitory effects on cell proliferation were assessed using the CCK-8 assay,colony formation assay,and EdU staining.The impact on cell migration and inva-sion was evaluated through wound-healing assays and Transwell migration and invasion assays.was employed toThe effects of CRT0066101 on the phosphorylation levels of PKD and key proteins in the downstream PI3K/AKT signaling pathway were analyzed using Western blot.Additionally,the drug's regulatory effects on apoptosis and autophagy in HCC cells were examined using Western blot,flow cytometry,and the mRFP-GFP-LC3 dual-fluorescence reporter system.Results CRT0066101 significantly inhibited the pro-liferation,migration and invasion of HCC cells.West-ern blotting results demonstrated that CRT0066101 dose-dependently suppressed the phosphorylation of PKD family proteins and downregulated the activation of the PI3K/AKT signaling pathway.Furthermore,CRT0066101 treatment upregulated the expression of the pro-apoptotic protein Bax while downregulating the anti-apoptotic protein Bcl-2.It also markedly increased the expression levels of autophagy marker proteins Bec-lin-1 and LC3B-Ⅱ,suggesting that the drug simulta-neously induced apoptosis and autophagy in HCC cells.Conclusions CRT0066101 specifically inhibits PKD activity,blocks the PI3K/AKT signaling path-way,suppresses HCC cell proliferation and metastasis,and induces apoptosis and autophagy.These findings indicate that CRT0066101 is a promising small-mole-cule inhibitor for targeted HCC therapy with potential clinical applications.

OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.

Objective To investigate the impact of county-level"Unified ECG Network"construction on the treatment efficiency and clinical outcomes of patients with acute ST-segment elevation myocardial infarction(STEMI).Methods A retrospective analysis was conducted on the clinical data of STEMI patients from Beichuan County and Yanting County in Mianyang City,and Jiange County in Guangyuan City,Sichuan Province,during the 18 months before(128 cases)and 18 months after(187 cases)the establishment of the"Unified ECG Network."Differences in demographic characteristics,treatment efficiency,therapeutic methods,and clinical outcomes between the two groups were compared.Results There was no statistically significant difference in general demographic characteristics between the two groups(all P＞0.05).Compared with the pre-construction group,the post-construction group showed significantly shorter times in initial ECG completion[5(3,7)min vs.6(4,8)min],initial ECG diagnosis[3(2,4)min vs.5(2,6)min],first medical contact to preliminary diagnosis[10(9,12)min vs.13(11,15)min],network hospital door-in-door-out time[21(19,23)min vs.26(23,30)min],and first medical contact to wire-crossing time[(94.82±11.87)min vs.(107.97±18.39)min](allP＜0.001).The proportion of patients bypassing the emergency department and coronary care unit significantly increased(64.17％vs.32.81％,P＜0.001).The proportion of patients undergoing emergency percutaneous coronary intervention significantly increased(72.73％vs.51.56％,P＜0.001),while the proportions of thrombolytic therapy and non-reperfusion therapy significantly decreased(both P＜0.05).Additionally,in-hospital mortality rate,Killip class≥Ⅱ proportion,incidence of major adverse cardiovascular events,and average length of hospital stay were all significantly reduced(all P＜0.05).There were no statistically significant differences among the three county-level chest pain centers in terms of major treatment efficiency,therapeutic strategies,or clinical outcomes(all P＞0.05).Conclusions The construction of the county-level"Unified ECG Network"can significantly improve the treatment efficiency of STEMI patients,optimize reperfusion therapy strategies,improve clinical outcomes,and demonstrate substantial clinical promotion value.

Objective To develop and validate a fast Monte Carlo(MC)-based patient-specific quality assurance(PSQA)tool using the treatment log files that is suitable to be used in the online adaptive radiotherapy for pencil beam scanning proton therapy(PBSPT-ART).Methods The proposed tool first used the delivery log file of a PBSPT plan to reversely reconstruct the PBSPT(rPBSPT)plan,and then used an in-house developed graphic processing unit(GPU)-accelerated virtual particle MC(VPMC)dose engine to calculate the dose distribution of the rPBSPT plan.The rPBSPT dose calculated by VPMC was then compared to the rPBSPT dose calculated by another independent MC dose engine(MCsquare),using 3D gamma analysis to verify the accuracy of VPMC calculation.As a demonstration of the feasibility of developed log file-based PSQA,the VPMC calculated dose of the rPBSPT plan was compared to the pre-delivery second check dose of the corresponding PBSPT plan calculated by MCsquare,using 3D gamma analysis.3D gamma analysis employes a criterion of 2 mm/2%/10%.Twenty patients with different disease sites were representatively selected to validate the efficiency and accuracy of the tool.Results The average calculation time of a rPBSPT plan by VPMC was(5.88±4.00)s in the accuracy verification.Compared to MCsquare,the passing rate of the 3D gamma analysis was 99.47%±0.72%.In the proposed PSQA tool demonstration,the passing rate of comparing the VPMC calculated rPBSPT dose to MCsquare calculated second check dose of the corresponding PBSPT plan was 98.91%±0.92%.Conclusion The accuracy and efficiency of the tool can meet the requirements of PSQA in the online PBSPT-ART workflow.

AIM:To investigate the mechanism of Qianggu-Kangshu formula(QGKSF)in the treatment of rheumatoid arthritis(RA)bone destruction based on network pharmacology,molecular docking,and cell experiments.METHODS:The main active ingredients and potential targets of QGKSF were obtained through TCMSP database and litera-ture search.OMIM database and GeneCards database were used to search the targets related to RA bone destruction,and Venny 2.1.0 was employed to screen the intersected targets of QGKSF and RA bone destruction.The protein-protein inter-action network of potential intersected targets was constructed by STRING database,and topological analysis was carried out by Cytoscape software to screen core targets.The screened targets of QGKSF related to RA bone destruction were evalu-ated through the Metascape system.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology were applied to complete enrichment analysis.AutoDock and PyMOL software was used to carry out molecular docking between the core components and the core proteins.Mouse RAW264.7 macrophages were cultured in vitro,and the cell viability was de-tected by CCK-8 assay.The number of tartrate-resistant acid phosphatase(TRAP)positive multinucleated cells in each group was calculated by TRAP staining.The enzyme activity of the cells was evaluated by determination of TRAP activity.F-actin ring formation was detected by phalloidin staining.Western blot analysis was conducted to detect the protein levels of nuclear factor of activated T cells 1(NFATc1),TRAP,cathepsin K(CTSK),c-Fos,matrix metalloproteinase 9(MMP9),osteoprotegerin(OPG),receptor activator of nuclear factor κB(RANK),RANK ligand(RANKL)and phos-phorylated protein kinase B(p-AKT).RESULTS:A total of 136 active ingredients and 126 targets were selected corre-lated with QGKSF,whil 207 intersected targets of QGKSF and RA bone destruction were screened,175 of which were core targets.There were 199 pathways obtained by GO enrichment,and 20 pathways related to osteoclasts were screened out,including phosphatidylinoinosiol 3-kinase/AKT signaling pathway and osteoclast differentiation,etc.The results of TRAP staining,TRAP enzyme activity determination and phalloidin staining showed less positive cell formation,de-creased enzyme activity and decreased F-actin ring formation in QGKSF and methotrexate(MTX)groups compared with model group.Western blot results showed that compared with model group,the protein levels of NFATc1,TRAP,CTSK,c-Fos,MMP9,p-AKT,RANK and RANKL were decreased(P＜0.05),while the expression of OPG protein was in-creased(P＜0.05)in QGKSF and MTX groups.CONCLUSION:Treatment with QGKSF inhibits RANKL-induced dif-ferentiation of RAW264.7 cells into osteoclasts possibly by inhibiting the over-differentiation of osteoclasts via regulating RANKL/RANK/OPG signaling pathway.

Objective::To compare the analgesic effects of ibuprofen administered orally via two modes combined with a conventional, patient-controlled intravenous analgesia pump on maternal pain after cesarean section (CS).Methods::This prospective, randomized, controlled study enrolled females who underwent CS from August 2022 to August 2023 at Peking University First Hospital, Beijing, China. Participants were randomly assigned to either an as-needed ibuprofen group (300 mg orally upon request) or a scheduled ibuprofen group (300 mg every 12 hours for 48 hours). The primary outcomes assessed were postoperative pain levels using the Wong-Baker Faces Pain Scale-Revised and cumulative oxycodone consumption at multiple time points up to 48 hours post-delivery. Secondary outcomes included recovery parameters (time to first flatus, ambulation, and lactation initiation), patient satisfaction with pain control, and postpartum depression scores evaluated by the Edinburgh Postnatal Depression Scale on postoperative day 3. Normally distributed data analyzed with t-tests; non-normal data with Mann-Whitney U tests; categorical variables with chi-square or Fisher’s exact tests (SPSS 26.0, P < 0.05). Results::After excluding 61 non-eligible cases, 339 patients were included (171 as-needed vs. 168 scheduled). The scheduled group showed significantly better pain control at 12 hours (4.00 (2.00-5.50) vs. 4.00 (4.00-6.00), P < 0.001), 24 hours (4.00 (2.00-4.00) vs. 4.00 (2.00-6.00), P < 0.001), and 36 hours (2.00 (2.00-4.00) vs. 4.00 (2.00-4.00), P < 0.001), and 48 hours (2.00 (2.00-4.00) vs. 2.00 (2.00-4.00), P = 0.004) post-delivery and lower levels of oxycodone consumption at 36 hours (10.20 (8.20-13.35) vs. 11.00 (8.80-14.40), P = 0.042) and 48 hours (12.40 (10.40-15.95) vs. 13.80 (11.00-16.00), P = 0.020) postpartum compared with those in the as-needed group. Additionally, the time to the return of bowel movements was shorter in the scheduled group than in the as-needed group (23.50 (16.94, 31.47) vs. 27.00 (19.88, 35.97), P = 0.004). Differences in post-delivery ambulation, lactation initiation, satisfaction levels, and depression scores were not significantly different between the two groups. Conclusion::The results of this study promote the use of ibuprofen (scheduled oral administration) combined with a conventional, patient-controlled intravenous analgesia pump for achieving better post-CS pain control than an as-needed dosage regimen.Registration::Chinese Clinical Trial Registry, ChiCTR2400082474.