Objective To explore the association between cardiorespiratory fitness and total atherosclerotic burden of brain-and heart-arteries in patients with ischemic stroke.Methods This study retrospectively enrolled consecutive patients with ischemic stroke of the large-artery atherosclerosis or small-artery occlusion subtypes accompanied by cervicocephalic atherosclerosis,who were admitted to the Department of Neurology,Xuanwu Hospital,Capital Medical University,between May 2023 and December 2024.General and clinical data,including age(＜60years,≥60years),sex,admission blood pressure,body mass index,history of hypertension,diabetes mellitus,hyperlipidemia,ischemic stroke,symptomatic coronary artery disease,smoking,alcohol consumption,and medication use(including antiplatelet drugs,anticoagulants,lipid-lowering drugs,antidiabetic drugs,antihypertensive drugs,and β-blockers),characteristics of ischemic stroke(including the duration of onset,ischemic side[left,right,bilateral],and ischemic location[anterior circulation,posterior circulation,bilateral circulation])were collected from all patients enrolled.On the morning after admission,fasting venous blood samples were collected to measure the levels of triglycerides,total cholesterol,low-density lipoprotein cholesterol,high-density lipoprotein cholesterol,fasting blood glucose,glycated hemoglobin,homocysteine,fibrinogen,D-dimer,C-reactive protein,high-sensitivity C-reactive protein,N-terminal pro-B-type natriuretic peptide,creatine kinase-MB,cardiac troponin T,and myoglobin.Transthoracic echocardiography was performed within 7 days after admission to evaluate cardiac function.Left ventricular ejection fraction and cardiac output were recorded,and cardiac index were calculated.Upon admission,neurological impairment,disability,motor function,and balance ability were assessed using the National Institutes of Health stroke scale(NIHSS),modified Rankin scale(mRS),Fugl-Meyer assessment(FMA),and Berg balance scale,respectively.The Saltin-Grimby physical activity level scale was used to evaluate the intensity of habitual physical activity prior to stroke onset.Cardiopulmonary exercise testing was performed within 7 days after admission to assess cardiopulmonary fitness.During the test,peak oxygen uptake(VO2peak),percentage of predicted VO2peak(VO2peak%pred),and metabolic equivalent of task(MET)were recorded.Based on the minimum oxygen uptake required for independent living(15 ml/[kg·min]),VO2peak was classified into low VO2peak(＜15 ml/[kg·min])and high VO2peak(≥15 ml/[kg·min]).With VO2peak%pred＜0.60 as the cutoff,VO2peak%pred was divided into low VO2peak%pred(＜0.60)and high VO2peak%pred(≥0.60).MET were categorized into low MET(＜mean MET)and high MET(≥mean MET)according to the mean value of MET in this study population.Imaging data of cardio-cerebral arteries were obtained by simultaneous CT arteriography within 7 days after admission.Total atherosclerotic burden of brain-and heart-supplying arteries(TAB-BHAs)was used to quantitatively evaluate the overall degree of cardio-cerebral atherosclerosis.And patients were stratified into high TAB-BHAs(≥4 points)and low TAB-BHAs(＜4 points)groups.The differences in general and clinical characteristics,as well as cardiopulmonary fitness indices,were assessed among different groups of TAB-BHAs.Spearman' s correlation was used in the preliminary analysis of the relationship between cardiorespiratory fitness indicators and TAB-BHAs.Multivariate Logistic regression was conducted with VO2peak,low VO2peak%pred,and low METs as independent variables,and high TAB-BHAs as the dependent variable to assess the association between cardiopulmonary fitness and TAB-BHAs.Covariates were selected through regressions stratified by age≥60 years,male,the least absolute shrinkage and selection operator(LASSO),and the variables exhibit statistically significant differences between the low and high TAB-BHAs groups,respectively.Results A total of 104 patients with ischemic stroke were enrolled.Among all patients enrolled,60patients(with ages ranging from 30 to 72years and a mean age of[54±11]years)met the inclusion and exclusion criteria,and were ultimately included.Amid the 60 patients,52 were male and 8 were female.There were 20 patients(33.3%)with low VO2peak,28 patients(46.7%)with low VO2peak%pred,and 31 patients(51.7%)with low MET(MET＜5.0).45 patients were classified into the low TAB-BHAs group and 15 into the high TAB-BHAs group.(1)Compared with the low TAB-BHAs group,patients in the high TAB-BHAs group were significantly older and had a higher proportion of patients aged≥60 years,they also showed significantly higher fibrinogen levels and a history of antihypertensive drug use(all P＜0.05).No significant differences were observed in other general or clinical characteristics between the two groups(all P＞0.05).Regarding cardiopulmonary fitness parameters,patients in the high TAB-BHAs group had lower VO2peak and MET levels,and higher proportions of patients with low VO2peak,low VO2peak%pred,and low MET compared with those in the low TAB-BHAs group(all P＜0.05).(2)Based on the Spearman's correlation analysis,VO2peak(r=-0.298,P=0.021),VO2peak%pred(r=-0.305,P=0.018),MET(r=-0.303,P=0.018)all exhibited negative correlations between and TAB-BHAs.(3)LASSO regression was used to identify the most pertinent variables for high TAB-BHAs,including low VO2peak,low VO2peak%pred,and fibrinogen.Multivariate Logistic regression analysis was then performed with low VO2peak,low VO2peak%pred,and low MET as independent variables and high TAB-BHAs as the dependent variable.The results showed that after adjusting for age≥60 years,male,and fibrinogen,both low VO2peak(adjusted OR,4.420,95%CI 1.052-18.573,P=0.042)and low VO2peak%pred(adjusted OR,7.423,95%CI 1.411-39.046,P=0.018)were independently associated with high TAB-BHAs in ischemic stroke patients.After further adjusting for variables that differed among high and low TAB-BHAs groups(including age≥60 years,male,fibrinogen,and history of using antihypertensive drugs),multivariate Logistic regression showed that low VO2peak%pred remained independently associated with high TAB-BHAs in ischemic stroke patients(adjusted OR,6.347,95%CI 1.170-34.418,P=0.032).Conclusion Poor cardiopulmonary fitness might be associated with high degree of overall cardio-cerebral atherosclerosis,suggesting cardiorespiratory fitness as a potential correlated indicator that could provide clues for improving early screening of patients with heavy cardio-cerebral atherosclerotic burden and for exploring the underlying mechanisms.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective To explore the association between cardiorespiratory fitness and total atherosclerotic burden of brain-and heart-arteries in patients with ischemic stroke.Methods This study retrospectively enrolled consecutive patients with ischemic stroke of the large-artery atherosclerosis or small-artery occlusion subtypes accompanied by cervicocephalic atherosclerosis,who were admitted to the Department of Neurology,Xuanwu Hospital,Capital Medical University,between May 2023 and December 2024.General and clinical data,including age(＜60years,≥60years),sex,admission blood pressure,body mass index,history of hypertension,diabetes mellitus,hyperlipidemia,ischemic stroke,symptomatic coronary artery disease,smoking,alcohol consumption,and medication use(including antiplatelet drugs,anticoagulants,lipid-lowering drugs,antidiabetic drugs,antihypertensive drugs,and β-blockers),characteristics of ischemic stroke(including the duration of onset,ischemic side[left,right,bilateral],and ischemic location[anterior circulation,posterior circulation,bilateral circulation])were collected from all patients enrolled.On the morning after admission,fasting venous blood samples were collected to measure the levels of triglycerides,total cholesterol,low-density lipoprotein cholesterol,high-density lipoprotein cholesterol,fasting blood glucose,glycated hemoglobin,homocysteine,fibrinogen,D-dimer,C-reactive protein,high-sensitivity C-reactive protein,N-terminal pro-B-type natriuretic peptide,creatine kinase-MB,cardiac troponin T,and myoglobin.Transthoracic echocardiography was performed within 7 days after admission to evaluate cardiac function.Left ventricular ejection fraction and cardiac output were recorded,and cardiac index were calculated.Upon admission,neurological impairment,disability,motor function,and balance ability were assessed using the National Institutes of Health stroke scale(NIHSS),modified Rankin scale(mRS),Fugl-Meyer assessment(FMA),and Berg balance scale,respectively.The Saltin-Grimby physical activity level scale was used to evaluate the intensity of habitual physical activity prior to stroke onset.Cardiopulmonary exercise testing was performed within 7 days after admission to assess cardiopulmonary fitness.During the test,peak oxygen uptake(VO2peak),percentage of predicted VO2peak(VO2peak%pred),and metabolic equivalent of task(MET)were recorded.Based on the minimum oxygen uptake required for independent living(15 ml/[kg·min]),VO2peak was classified into low VO2peak(＜15 ml/[kg·min])and high VO2peak(≥15 ml/[kg·min]).With VO2peak%pred＜0.60 as the cutoff,VO2peak%pred was divided into low VO2peak%pred(＜0.60)and high VO2peak%pred(≥0.60).MET were categorized into low MET(＜mean MET)and high MET(≥mean MET)according to the mean value of MET in this study population.Imaging data of cardio-cerebral arteries were obtained by simultaneous CT arteriography within 7 days after admission.Total atherosclerotic burden of brain-and heart-supplying arteries(TAB-BHAs)was used to quantitatively evaluate the overall degree of cardio-cerebral atherosclerosis.And patients were stratified into high TAB-BHAs(≥4 points)and low TAB-BHAs(＜4 points)groups.The differences in general and clinical characteristics,as well as cardiopulmonary fitness indices,were assessed among different groups of TAB-BHAs.Spearman' s correlation was used in the preliminary analysis of the relationship between cardiorespiratory fitness indicators and TAB-BHAs.Multivariate Logistic regression was conducted with VO2peak,low VO2peak%pred,and low METs as independent variables,and high TAB-BHAs as the dependent variable to assess the association between cardiopulmonary fitness and TAB-BHAs.Covariates were selected through regressions stratified by age≥60 years,male,the least absolute shrinkage and selection operator(LASSO),and the variables exhibit statistically significant differences between the low and high TAB-BHAs groups,respectively.Results A total of 104 patients with ischemic stroke were enrolled.Among all patients enrolled,60patients(with ages ranging from 30 to 72years and a mean age of[54±11]years)met the inclusion and exclusion criteria,and were ultimately included.Amid the 60 patients,52 were male and 8 were female.There were 20 patients(33.3%)with low VO2peak,28 patients(46.7%)with low VO2peak%pred,and 31 patients(51.7%)with low MET(MET＜5.0).45 patients were classified into the low TAB-BHAs group and 15 into the high TAB-BHAs group.(1)Compared with the low TAB-BHAs group,patients in the high TAB-BHAs group were significantly older and had a higher proportion of patients aged≥60 years,they also showed significantly higher fibrinogen levels and a history of antihypertensive drug use(all P＜0.05).No significant differences were observed in other general or clinical characteristics between the two groups(all P＞0.05).Regarding cardiopulmonary fitness parameters,patients in the high TAB-BHAs group had lower VO2peak and MET levels,and higher proportions of patients with low VO2peak,low VO2peak%pred,and low MET compared with those in the low TAB-BHAs group(all P＜0.05).(2)Based on the Spearman's correlation analysis,VO2peak(r=-0.298,P=0.021),VO2peak%pred(r=-0.305,P=0.018),MET(r=-0.303,P=0.018)all exhibited negative correlations between and TAB-BHAs.(3)LASSO regression was used to identify the most pertinent variables for high TAB-BHAs,including low VO2peak,low VO2peak%pred,and fibrinogen.Multivariate Logistic regression analysis was then performed with low VO2peak,low VO2peak%pred,and low MET as independent variables and high TAB-BHAs as the dependent variable.The results showed that after adjusting for age≥60 years,male,and fibrinogen,both low VO2peak(adjusted OR,4.420,95%CI 1.052-18.573,P=0.042)and low VO2peak%pred(adjusted OR,7.423,95%CI 1.411-39.046,P=0.018)were independently associated with high TAB-BHAs in ischemic stroke patients.After further adjusting for variables that differed among high and low TAB-BHAs groups(including age≥60 years,male,fibrinogen,and history of using antihypertensive drugs),multivariate Logistic regression showed that low VO2peak%pred remained independently associated with high TAB-BHAs in ischemic stroke patients(adjusted OR,6.347,95%CI 1.170-34.418,P=0.032).Conclusion Poor cardiopulmonary fitness might be associated with high degree of overall cardio-cerebral atherosclerosis,suggesting cardiorespiratory fitness as a potential correlated indicator that could provide clues for improving early screening of patients with heavy cardio-cerebral atherosclerotic burden and for exploring the underlying mechanisms.

Endovascular aneurysm repair (EVAR) is a key treatment for abdominal aortic aneurysms (AAA),but type Ⅱ endoleak (T2EL) is one of the most common postoperative complications. T2EL mainly arises from retrograde blood flow from collateral vessels such as the inferior mesenteric artery (IMA) and lumbar arteries (LA),which continue to perfuse the aneurysm sac. Although T2EL may lead to aneurysm sac enlargement or even rupture,its overall clinical impact is relatively minor,and most cases can be effectively managed through observation or minimally invasive interventions. Currently,the efficacy of prophylactic embolization of the IMA and LA in reducing the incidence of T2EL remains uncertain and may increase operative time,radiation exposure,and the risk of associated complications. More importantly,authoritative guidelines,both domestic and international,do not recommend routine prophylactic embolization of the IMA and LA during EVAR. Whether prophylactic embolization should be performed in high-risk patients for T2EL remains controversial,with most studies in this area being small-sample or single-center retrospective analyses,offering limited evidence quality. This article analyzes the harm of T2EL,the effectiveness of prophylactic embolization,and relevant guideline recommendations based on existing research,aiming to provide a reference for the individualized management of T2EL following EVAR.

Objective To investigate the safety and feasibility of peripheral vascular intervention via the ipsilateral transulnar access(TUA)due to failure of transradial access(TRA)puncture.Methods The clinical data of 2546 peripheral vascular interventions via TRA,which were performed at authors'hospital between January 2019 and December 2021,were retrospectively analyzed.Among the 2546 interventions,TRA puncture failed in 37 procedures,and in 27 of these patients the ipsilateral TUA puncture had to be adopted.The puncture success rate,surgical success rate and puncture approach-related complications of TUA of the 27 patients receiving ipsilateral TUA puncture were analyzed.Results The success rate of ipsilateral TUA puncture after TRA puncture failed was 96.3％(26/27),and in one patient transfemoral access(TFA)puncture had to be substituted because of the ulnar artery spasm.The total success rate of interventional procedures was 96.3％(26/27).No serious complications occurred,and the incidence of minor complications was 19.2％(5/26).Conclusion Preliminary results indicate that for the experienced TRA operators,using ipsilateral TUA puncture due to failure of TRA puncture is a safe and feasible strategy choice.

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.

Endovascular aneurysm repair (EVAR) is a key treatment for abdominal aortic aneurysms (AAA),but type Ⅱ endoleak (T2EL) is one of the most common postoperative complications. T2EL mainly arises from retrograde blood flow from collateral vessels such as the inferior mesenteric artery (IMA) and lumbar arteries (LA),which continue to perfuse the aneurysm sac. Although T2EL may lead to aneurysm sac enlargement or even rupture,its overall clinical impact is relatively minor,and most cases can be effectively managed through observation or minimally invasive interventions. Currently,the efficacy of prophylactic embolization of the IMA and LA in reducing the incidence of T2EL remains uncertain and may increase operative time,radiation exposure,and the risk of associated complications. More importantly,authoritative guidelines,both domestic and international,do not recommend routine prophylactic embolization of the IMA and LA during EVAR. Whether prophylactic embolization should be performed in high-risk patients for T2EL remains controversial,with most studies in this area being small-sample or single-center retrospective analyses,offering limited evidence quality. This article analyzes the harm of T2EL,the effectiveness of prophylactic embolization,and relevant guideline recommendations based on existing research,aiming to provide a reference for the individualized management of T2EL following EVAR.

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease with unclear etiology and limited treatment options. The median survival time for IPF patients is approximately 2-3 years and there is no effective intervention to treat IPF other than lung transplantation. As important components of lung tissue, endothelial cells (ECs) are associated with pulmonary diseases. However, the role of endothelial dysfunction in pulmonary fibrosis (PF) is incompletely understood. Sphingosine-1-phosphate receptor 1 (S1PR1) is a G protein-coupled receptor highly expressed in lung ECs. Its expression is markedly reduced in patients with IPF. Herein, we generated an endothelial-conditional S1pr1 knockout mouse model which exhibited inflammation and fibrosis with or without bleomycin (BLM) challenge. Selective activation of S1PR1 with an S1PR1 agonist, IMMH002, exerted a potent therapeutic effect in mice with bleomycin-induced fibrosis by protecting the integrity of the endothelial barrier. These results suggest that S1PR1 might be a promising drug target for IPF therapy.