Objective Size distribution is an important biophysical property of extracellular vesicles(EVs).EVs include small EVs(s-EVs)and large EVs(l-EVs)by size.Differential ultracentrifugation(dUC)is widely used to separate E...Objective Size distribution is an important biophysical property of extracellular vesicles(EVs).EVs include small EVs(s-EVs)and large EVs(l-EVs)by size.Differential ultracentrifugation(dUC)is widely used to separate EVs from biofluids,but it can precipitate large impurity particles.Dynamic light scattering(DLS)is a simple and fast method for analyzing the size distribution of EVs.However,this approach is nonideal for heterogeneous and polydisperse samples since a small quantity of large impurity particles can markedly disturb the DLS results.Here,we developed a simple method to improve the reliability of DLS measurements.Methods Plasma was obtained from 13 volunteers.The plasma was first processed by dUC to obtain crude l-EVs.The crude l-EVs were filtered with syringe filters(pore size of 1μm and membrane material of hydrophilic polyvinylidene fluoride(PVDF))to remove large impurity particles from l-EVs.The size distributions of the crude l-EVs and filtered l-EVs were measured via DLS.Results After the samples were filtered,the coefficients of variation of the hydrodynamic radius and Peak 1 intensity of the filtered l-EVs decreased from 20.39%(12.76–28.96%)and 20.44%(14.58–28.32%)to 3.05%(1.79–4.72%)and 3.43%(1.76–5.88%),respectively,compared with those of the crude l-EVs.Conclusion These findings suggest that filtration can effectively separate circulating l-EVs in plasma to remove large impurity particles and make samples suitable for characterization by DLS.Our findings provide a simple method to improve precision via DLS to measure the size distribution of EVs.展开更多
Growing evidence has demonstrated exercise as an effective way to promote cardiovascular health and protect against cardiovascular diseases However,the underlying mechanisms of the beneficial effects of exercise have ...Growing evidence has demonstrated exercise as an effective way to promote cardiovascular health and protect against cardiovascular diseases However,the underlying mechanisms of the beneficial effects of exercise have yet to be elucidated.Animal exercise studies are widely used to investigate the key mechanisms of exercise-induced cardiovascular protection.However,standardized procedures and well-established evaluation indicators for animal exercise models are needed to guide researchers in carrying out effective,high-quality animal studies using exercise to prevent and treat cardiovascular diseases.In our review,we present the commonly used animal exercise models in cardiovascular research and propose a set of standard procedures for exercise training,emphasizing the appropriate measurements and analysis in these chronic exercise models.We also provide recommendations for optimal design of animal exercise studies in cardiovascular research,including the choice of exercise models,control of exercise protocols,exercise at different stages of disease,and other considerations,such as age,sex,and genetic background.We hope that this position paper will promote basic research on exercise-induced cardiovascular protection and pave the way for successful translation of exercise studies from bench to bedside in the prevention and treatment of cardiovascular diseases.展开更多
Acute heart failure(AHF)is a severe complication after cardiac surgery with cardiopulmonary bypass(CPB).Although some AHF biomarkers have been used in clinic,they have limitations when applied in the prediction and di...Acute heart failure(AHF)is a severe complication after cardiac surgery with cardiopulmonary bypass(CPB).Although some AHF biomarkers have been used in clinic,they have limitations when applied in the prediction and diagnosis of AHF after cardiac surgery with CPB,and there are still no effective and specific biomarkers.We and other researchers have shown that circulating microparticles(MPs)increased in a variety of cardiovascular diseases.However,whether the concentration of circulating MPs could be a new biomarker for AHF after cardiac surgery remains unknown.Here,90 patients undergoing cardiac surgery with CPB and 45 healthy subjects were enrolled.Patients were assigned into AHF(n=14)or non-AHF(n=76)group according to the diagnosis criteria of AHF.The concentrations of circulating MPs were determined before,as well as 12 h and 3 days after operation with nanoparticle tracking analysis technique.MPs concentrations in patients before surgery were significantly higher than those of healthy subjects.Plasma levels of MPs were significantly elevated at 12 h after surgery in patients with AHF,but not in those without AHF,and the circulating MPs concentrations at 12 h after surgery were higher in AHF group compared with non-AHF group.Logistic regression analysis indicated that MPs concentration at postoperative 12 h was an independent risk factor for AHF.The area under receiver operating characteristic curve for MPs concentration at postoperative 12 h was 0.87 and the best cut-off value is 5.20×10~8 particles mL~(–1)with a sensitivity of 93%and a specificity of 70%.These data suggested that the concentration of circulating MPs might be a new biomarker for the occurrence of AHF after cardiac surgery with CPB.展开更多
Electrospinning is a popular and effective method of producing porous nanofibers with a large surface area,superior physical and chemical properties,and a controllable pore size.Owing to these properties,electrospun n...Electrospinning is a popular and effective method of producing porous nanofibers with a large surface area,superior physical and chemical properties,and a controllable pore size.Owing to these properties,electrospun nanofibers can mimic the extracellular matrix and some human tissue structures,based on the fiber configuration.Consequently,the application of electrospun nanofibers as biomaterials,varying from two-dimensional(2D)wound dressings to three-dimensional(3D)tissue engineering scaffolds,has increased rapidly in recent years.Nanofibers can either be uniform fiber strands or coaxial drug carriers,and their overall structure varies from random mesh-like mats to aligned or gradient scaffolds.In addition,the pore size of the fibers can be adjusted or the fibers can be loaded with disparate medicines to provide different functions.This review discusses the various structures and applications of 2D fiber mats and 3D nanofibrous scaffolds made up of different one-dimensional(1D)fibers in tissue engineering.In particular,we focus on the improvements made in recent years,especially in the fields of wound healing,angiogenesis,and tissue regeneration.展开更多
Diabetic foot ulcers(DFUs)are a serious vascular disease.Currently,no effective methods are available for treating DFUs.Pro-protein convertase subtilisin/kexin type 9(PCSK9)regulates lipid levels to promote atheroscle...Diabetic foot ulcers(DFUs)are a serious vascular disease.Currently,no effective methods are available for treating DFUs.Pro-protein convertase subtilisin/kexin type 9(PCSK9)regulates lipid levels to promote atherosclerosis.However,the role of PCSK9 in DFUs remains unclear.In this study,we found that the expression of PCSK9 in endothelial cells(ECs)increased significantly under high glucose(HG)stimulation and in diabetic plasma and vessels.Specifically,PCSK9 promotes the E3 ubiquitin-protein ligase NEDD4 binding to vascular endothelial growth factor receptor 2(VEGFR2),which led to the ubiquitination of VEGFR2,resulting in its degradation and downregulation in ECs.Furthermore,PCSK9 suppresses the expression and activation of AKT,endothelial nitric oxide synthase(e NOS),and ERK1/2,leading to decreased nitric oxide(NO)production and increased superoxide anion(O_(2)^(·-))generation,which impairs vascular endothelial function and angiogenesis.Importantly,using evolocumab to limit the increase in PCSK9 expression blocked the HG-induced inhibition of NO production and the increase in O_(2)^(·-)production,as well as inhibited the phosphorylation and expression of AKT,e NOS,and ERK1/2.Moreover,evolocumab improved vascular endothelial function and angiogenesis,and promoted wound healing in diabetes.Our findings suggest that targeting PCSK9 is a novel therapeutic approach for treating DFUs.展开更多
Cardiopulmonary bypass has been speculated to elicit systemic inflammation to initiate acute lung injury(ALI), including acute respiratory distress syndrome(ARDS), in patients after cardiac surgery. We previously foun...Cardiopulmonary bypass has been speculated to elicit systemic inflammation to initiate acute lung injury(ALI), including acute respiratory distress syndrome(ARDS), in patients after cardiac surgery. We previously found that post-operative patients showed an increase in endothelial cell-derived extracellular vesicles(eEVs) with components of coagulation and acute inflammatory responses. However, the mechanism underlying the onset of ALI owing to the release of e EVs after cardiopulmonary bypass, remains unclear. Plasma plasminogenactivated inhibitor-1(PAI-1) and eEV levels were measured in patients with cardiopulmonary bypass. Endothelial cells and mice(C57BL/6,Toll-like receptor 4 knockout(TLR4^(-/-))) and inducible nitric oxide synthase knockout(iNOS^(-/-)) were challenged with eEVs isolated from PAI-1-stimulated endothelial cells. Plasma PAI-1 and eEVs were remarkably enhanced after cardiopulmonary bypass. Plasma PAI-1 elevation was positively correlated with the increase in eEVs. The increase in plasma PAI-1 and eEV levels was associated with post-operative ARDS. The eEVs derived from PAI-1-stimulated endothelial cells could recognize TLR4 to stimulate a downstream signaling cascade identified as the Janus kinase 2/3(JAK2/3)-signal transducer and activator of transcription 3(STAT3)-interferon regulatory factor 1(IRF-1)pathway, along with i NOS induction, and cytokine/chemokine production in vascular endothelial cells and C57BL/6 mice, ultimately contributing to ALI. ALI could be attenuated by JAK2/3 or STAT3 inhibitors(AG490 or S3I-201, respectively), and was relieved in TLR4-/-and iNOS-/-mice. eEVs activate the TLR4/JAK3/STAT3/IRF-1 signaling pathway to induce ALI/ARDS by delivering follistatin-like protein 1(FSTL1), and FSTL1 knockdown in eEVs alleviates eEV-induced ALI/ARDS. Our data thus demonstrate that cardiopulmonary bypass may increase plasma PAI-1 levels to induce FSTL1-enriched eEVs, which target the TLR4-mediated JAK2/3/STAT3/IRF-1 signaling cascade and form a positive feedback loop, leading to ALI/ARDS after cardiac surgery. Our findings provide new insight into the molecular mechanisms and therapeutic targets for ALI/ARDS after cardiac surgery.展开更多
Chronic heart failure is the end stage of cardiac diseases.With a high prevalence and a high mortality rate worldwide,chronic heart failure is one of the heaviest health-related burdens.In addition to the standard neu...Chronic heart failure is the end stage of cardiac diseases.With a high prevalence and a high mortality rate worldwide,chronic heart failure is one of the heaviest health-related burdens.In addition to the standard neurohormonal blockade therapy,several medications have been developed for chronic heart failure treatment,but the population-wide improvement in chronic heart failure prognosis over time has been modest,and novel therapies are still needed.展开更多
Wound ulceration caused by diabetes is a typical chronic wound wherein healing the local tissue is difficult due to lack of blood vessels and tissue necrosis caused by the long-term accumulation of free radicals.Near-...Wound ulceration caused by diabetes is a typical chronic wound wherein healing the local tissue is difficult due to lack of blood vessels and tissue necrosis caused by the long-term accumulation of free radicals.Near-field electrospinning(NFES)is an innovative technology used to produce micro-nano-scaled,controllable sequencing fibers.In this study,we constructed a novel wound dressing based on the NFES polycaprolactone(PCL)fiber network and modified gelatin with methacrylic anhydride(GelMA)hydrogel to promote angiogenesis and the re-epithelialization of diabetic wounds.An angiogenic and antioxidant drug named deferoxamine(DFO)was encapsulated in a GelMA hydrogel to achieve a slow-release effect that is more suitable for chronic wounds.The cell adhesion experiment showed that the cells could attach to the fibers in the dressing group having a network of PCL fibers on the surface and grow along the direction of the fibers,which in turn,effectively regulates cell behavior from the physical structure.Additionally,the large pore size(~500μm)of the network allowed the cells to penetrate the pores and enter the surface of the hydrogel without being blocked out.Besides,the composite dressing had a notable effect on angiogenesis.Furthermore,antioxidation experiments confirmed that the DFO-loaded hydrogel exhibited antioxidant activity.Experimental animal models of diabetes showed that rats treated with the PCL-GelMA-DFO(PGD)hydrogel had faster ability of hemostasis,scab formation,and wound healing.In conclusion,the PGD hydrogel effectively promoted the repair of chronic wounds.展开更多
Normal high-density lipoprotein(nHDL)can induce angiogenesis in healthy individuals.However,HDL from patients with coronary artery disease undergoes various modifications,becomes dysfunctional(dHDL),and loses its abil...Normal high-density lipoprotein(nHDL)can induce angiogenesis in healthy individuals.However,HDL from patients with coronary artery disease undergoes various modifications,becomes dysfunctional(dHDL),and loses its ability to promote angiogenesis.Here,we identified a long non-coding RNA,HDRACA,that is involved in the regulation of angiogenesis by HDL.In this study,we showed that nHDL downregulates the expression of HDRACA in endothelial cells by activating WW domain-containing E3 ubiquitin protein ligase 2,which catalyzes the ubiquitination and subsequent degradation of its transcription factor,Kruppel-like factor 5,via sphingosine 1-phosphate(S1P)receptor 1.In contrast,dHDL with lower levels of S1P than nHDL were much less effective in decreasing the expression of HDRACA.HDRACA was able to bind to Ras-interacting protein 1(RAIN)to hinder the interaction between RAIN and vigilin,which led to an increase in the binding between the vigilin protein and proliferating cell nuclear antigen(PCNA)mRNA,resulting in a decrease in the expression of PCNA and inhibition of angiogenesis.The expression of human HDRACA in a hindlimb ischemia mouse model inhibited the recovery of angiogenesis.Taken together,these findings suggest that HDRACA is involved in the HDL regulation of angiogenesis,which nHDL inhibits the expression of HDRACA to induce angiogenesis,and that dHDL is much less effective in inhibiting HDRACA expression,which provides an explanation for the decreased ability of dHDL to stimulate angiogenesis.展开更多
Although VEGF-B was discovered as a VEGF-A homolog a long time ago,the angiogenic effect of VEGF-B remains poorly understood with limited and diverse findings from different groups.Notwithstanding,drugs that inhibit V...Although VEGF-B was discovered as a VEGF-A homolog a long time ago,the angiogenic effect of VEGF-B remains poorly understood with limited and diverse findings from different groups.Notwithstanding,drugs that inhibit VEGF-B together with other VEGF family members are being used to treat patients with various neovascular diseases.It is therefore critical to have a better understanding of the angiogenic effect of VEGF-B and the underlying mechanisms.Using comprehensive in vitro and in vivo methods and models,we reveal here for the first time an unexpected and surprising function of VEGF-B as an endogenous inhibitor of angiogenesis by inhibiting the FGF2/FGFR1 pathway when the latter is abundantly expressed.Mechanistically,we unveil that VEGF-B binds to FGFR1,induces FGFR1/VEGFR1 complex formation,and suppresses FGF2-induced Erk activation,and inhibits FGF2-driven angiogenesis and tumor growth.Our work uncovers a previously unrecognized novel function of VEGF-B in tethering the FGF2/FGFR1 pathway.Given the anti-angiogenic nature of VEGF-B under conditions of high FGF2/FGFR1 levels,caution is warranted when modulating VEGF-B activity to treat neovascular diseases.展开更多
We previously demonstrated that normal high-density lipoprotein(nHDL)can promote angiogenesis,whereas HDL from patients with coronary artery disease(d HDL)is dysfunctional and impairs angiogenesis.Autophagy plays a cr...We previously demonstrated that normal high-density lipoprotein(nHDL)can promote angiogenesis,whereas HDL from patients with coronary artery disease(d HDL)is dysfunctional and impairs angiogenesis.Autophagy plays a critical role in angiogenesis,and HDL regulates autophagy.However,it is unclear whether n HDL and d HDL regulate angiogenesis by affecting autophagy.Endothelial cells(ECs)were treated with n HDL and d HDL with or without an autophagy inhibitor.Autophagy,endothelial nitric oxide synthase(e NOS)expression,miRNA expression,nitric oxide(NO)production,superoxide anion(O2^(·-))generation,EC migration,and tube formation were evaluated.n HDL suppressed the expression of miR-181a-5p,which promotes autophagy and the expression of e NOS,resulting in NO production and the inhibition of O2^(·-)generation,and ultimately increasing in EC migration and tube formation.d HDL showed opposite effects compared to n HDL and ultimately inhibited EC migration and tube formation.We found that autophagy-related protein 5(ATG5)was a direct target of miR-181a-5p.ATG5 silencing or miR-181a-5p mimic inhibited n HDL-induced autophagy,e NOS expression,NO production,EC migration,tube formation,and enhanced O2^(·-)generation,whereas overexpression of ATG5 or miR-181a-5p inhibitor reversed the above effects of d HDL.ATG5 expression and angiogenesis were decreased in the ischemic lower limbs of hypercholesterolemic low-density lipoprotein receptor null(LDLr^(-/-))mice when compared to C57BL/6 mice.ATG5 overexpression improved angiogenesis in ischemic hypercholesterolemic LDLr^(-/-)mice.Taken together,nHDL was able to stimulate autophagy by suppressing miR-181a-5p,subsequently increasing e NOS expression,which generated NO and promoted angiogenesis.In contrast,d HDL inhibited angiogenesis,at least partially,by increasing miR-181a-5p expression,which decreased autophagy and e NOS expression,resulting in a decrease in NO production and an increase in O2^(·-)generation.Our findings reveal a novel mechanism by which HDL affects angiogenesis by regulating autophagy and provide a therapeutic target for d HDL-impaired angiogenesis.展开更多
Cardiac fibrosis is a pathological response characterized by excessive deposition of fibrous connective tissue within the heart.It typically occurs following cardiac injuries or diseases.However,the lack of suitable m...Cardiac fibrosis is a pathological response characterized by excessive deposition of fibrous connective tissue within the heart.It typically occurs following cardiac injuries or diseases.However,the lack of suitable models for disease modeling and high-throughput drug discovery has hindered the establishment of an effective treatments for cardiac fibrosis.The emergence and rapid progress of stem-cell and lineage reprogramming technology offer an unprecedented opportunity to develop an improved humanized and patient-specific model for studying cardiac fibrosis,providing a platform for screening potential drugs and synchronously elucidating the underlying molecular mechanisms.Furthermore,reprogramming cardiac fibroblasts into cardiomyocyte-like cells to reduce scar volume and induce myocardial tissue regeneration is a promising approach in treating cardiac fibrosis.In this review,we summarize the current advancements in stem cell technologies applied to study cardiac fibrosis and provide insights for future investigations into its mechanisms,drug discovery as well as therapy method.展开更多
Background and Aims:Increasing utilization of extended criteria donor leads to an increasing rate of early allograft failure after liver transplantation.However,consensus of definition of early allograft failure is la...Background and Aims:Increasing utilization of extended criteria donor leads to an increasing rate of early allograft failure after liver transplantation.However,consensus of definition of early allograft failure is lacking.Methods:A retrospective,multicenter study was performed to validate the Liver Graft Assessment Following Transplantation(L-GrAFT)risk model in a Chinese cohort of 942 adult patients undergoing primary liver transplantation at three Chinese centers.L-GrAFT(L-GrAFT7 and L-GrAFT10)was compared with existing models:the Early Allograft Failure Simplified Estimation(EASE)score,the model of early allograft function(MEAF),and the Early Allograft Dysfunction(EAD)model.Univariate and multivariate logistic regression were used to find risk factors of L-GrAFT high-risk group.Results:L-GrAFT7 had an area under the curve of 0.85 in predicting 90-day graft survival,significantly superior to MEAF[area under the curve(AUC=0.78,p=0.044)]and EAD(AUC=0.78,p=0.006),while there was no statistical significance between the predicting abilities of L-GrAFT7 and EASE(AUC=0.84,p>0.05).Furthermore,L-GrAFT7 maintains good predicting ability in the subgroup of high-donor risk index(DRI)cases(AUC=0.83 vs.MEAF,p=0.007 vs.EAD,p=0.014)and recipients of donors after cardiac death(AUC=0.92 vs.EAD,p<0.001).Through multivariate analysis,pretransplant bilirubin level,units of packed red blood cells,and the DRI score were selected as independent risk factors of a L-GrAFT7 high-risk group.Conclusions:The accuracy of L-GrAFT7 in predicting early allograft failure was validated in a Chinese multicenter cohort,indicating that it has the potential to become an accurate endpoint of clinical practice and transitional study of machine perfusion.展开更多
基金National Natural Science Foundation of China(No.82495171,92268202,82270485)National Key R&D Program of China(No.2021YFA0805100)+5 种基金Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(No.2023-PT320-03)Guangdong Basic and Applied Basic Research Foundation,China(No.2024A1515030041)The Science and Technology Planning Project of Guangdong Province,(No.2023B1212060018)Sun Yat-sen University Basic Research Funds(No.24qnpy354)Clinical Research 5010 Program(No.2014002)Program of National Key Clinical Specialties.
文摘Objective Size distribution is an important biophysical property of extracellular vesicles(EVs).EVs include small EVs(s-EVs)and large EVs(l-EVs)by size.Differential ultracentrifugation(dUC)is widely used to separate EVs from biofluids,but it can precipitate large impurity particles.Dynamic light scattering(DLS)is a simple and fast method for analyzing the size distribution of EVs.However,this approach is nonideal for heterogeneous and polydisperse samples since a small quantity of large impurity particles can markedly disturb the DLS results.Here,we developed a simple method to improve the reliability of DLS measurements.Methods Plasma was obtained from 13 volunteers.The plasma was first processed by dUC to obtain crude l-EVs.The crude l-EVs were filtered with syringe filters(pore size of 1μm and membrane material of hydrophilic polyvinylidene fluoride(PVDF))to remove large impurity particles from l-EVs.The size distributions of the crude l-EVs and filtered l-EVs were measured via DLS.Results After the samples were filtered,the coefficients of variation of the hydrodynamic radius and Peak 1 intensity of the filtered l-EVs decreased from 20.39%(12.76–28.96%)and 20.44%(14.58–28.32%)to 3.05%(1.79–4.72%)and 3.43%(1.76–5.88%),respectively,compared with those of the crude l-EVs.Conclusion These findings suggest that filtration can effectively separate circulating l-EVs in plasma to remove large impurity particles and make samples suitable for characterization by DLS.Our findings provide a simple method to improve precision via DLS to measure the size distribution of EVs.
基金This work was supported by grants from the National Key Research and Development Project(2020YFA0803800 to YB)National Natural Science Foundation of China(82020108002 and 81911540486 to JX,81772444 to LW,81772466 to RD)+2 种基金Innovation Program of Shanghai Municipal Education Commission(2017-01-07-00-09-E00042 to JX)Science and Technology Commission of Shanghai Municipality(18410722200 and 17010500100 to JX)“Dawn”Program of the Shanghai Education Commission(19SG34 to JX)。
文摘Growing evidence has demonstrated exercise as an effective way to promote cardiovascular health and protect against cardiovascular diseases However,the underlying mechanisms of the beneficial effects of exercise have yet to be elucidated.Animal exercise studies are widely used to investigate the key mechanisms of exercise-induced cardiovascular protection.However,standardized procedures and well-established evaluation indicators for animal exercise models are needed to guide researchers in carrying out effective,high-quality animal studies using exercise to prevent and treat cardiovascular diseases.In our review,we present the commonly used animal exercise models in cardiovascular research and propose a set of standard procedures for exercise training,emphasizing the appropriate measurements and analysis in these chronic exercise models.We also provide recommendations for optimal design of animal exercise studies in cardiovascular research,including the choice of exercise models,control of exercise protocols,exercise at different stages of disease,and other considerations,such as age,sex,and genetic background.We hope that this position paper will promote basic research on exercise-induced cardiovascular protection and pave the way for successful translation of exercise studies from bench to bedside in the prevention and treatment of cardiovascular diseases.
基金supported by the National Natural Science Foundation of China(81670392,81600382,81770241,81830013,81970363)the National Science Fund for Distinguished Young Scholars(81325001)+4 种基金the International Cooperation Project(2015DFA31070)from the Ministry of Science and Technology of Chinathe National Key Research and Development Program of China(2016YFC0903000)the Changjiang Scholars Program from the Ministry of Education of China,Guangdong Natural Science Fund Committee(2015A030312009)the Guangdong Pearl River Scholars Programthe Sun Yat-sen University Clinical Research 5010 Program(2014002)。
文摘Acute heart failure(AHF)is a severe complication after cardiac surgery with cardiopulmonary bypass(CPB).Although some AHF biomarkers have been used in clinic,they have limitations when applied in the prediction and diagnosis of AHF after cardiac surgery with CPB,and there are still no effective and specific biomarkers.We and other researchers have shown that circulating microparticles(MPs)increased in a variety of cardiovascular diseases.However,whether the concentration of circulating MPs could be a new biomarker for AHF after cardiac surgery remains unknown.Here,90 patients undergoing cardiac surgery with CPB and 45 healthy subjects were enrolled.Patients were assigned into AHF(n=14)or non-AHF(n=76)group according to the diagnosis criteria of AHF.The concentrations of circulating MPs were determined before,as well as 12 h and 3 days after operation with nanoparticle tracking analysis technique.MPs concentrations in patients before surgery were significantly higher than those of healthy subjects.Plasma levels of MPs were significantly elevated at 12 h after surgery in patients with AHF,but not in those without AHF,and the circulating MPs concentrations at 12 h after surgery were higher in AHF group compared with non-AHF group.Logistic regression analysis indicated that MPs concentration at postoperative 12 h was an independent risk factor for AHF.The area under receiver operating characteristic curve for MPs concentration at postoperative 12 h was 0.87 and the best cut-off value is 5.20×10~8 particles mL~(–1)with a sensitivity of 93%and a specificity of 70%.These data suggested that the concentration of circulating MPs might be a new biomarker for the occurrence of AHF after cardiac surgery with CPB.
基金the funding from Medical Scientific Research Foundation of Guangdong Province(No.A2021093)Science and Technology Planning Project of Shenzhen Municipality(No.YJ20180306174831458)+7 种基金Shenzhen Basic Research Project(No.JCYJ20190807155801657)National Science and Technology Major Project of the Ministry of Science and Technology of China(No.2018ZX10301402)Key International(Regional)Joint Research Program of China(No.5181001045)Guangdong Innovative and Entrepreneurial Research Team Program(No.2016ZT06S029)the National Natural Science Foundation of China(No.51973243)China Postdoctoral Science Foundation(No.2019M663246)the Fundamental Research Funds for the Central Universities(Nos.191gzd35 and 20ykpyl5)Guangdong Basic and Applied Basic Research Foundation(No.2019A1515110686).
文摘Electrospinning is a popular and effective method of producing porous nanofibers with a large surface area,superior physical and chemical properties,and a controllable pore size.Owing to these properties,electrospun nanofibers can mimic the extracellular matrix and some human tissue structures,based on the fiber configuration.Consequently,the application of electrospun nanofibers as biomaterials,varying from two-dimensional(2D)wound dressings to three-dimensional(3D)tissue engineering scaffolds,has increased rapidly in recent years.Nanofibers can either be uniform fiber strands or coaxial drug carriers,and their overall structure varies from random mesh-like mats to aligned or gradient scaffolds.In addition,the pore size of the fibers can be adjusted or the fibers can be loaded with disparate medicines to provide different functions.This review discusses the various structures and applications of 2D fiber mats and 3D nanofibrous scaffolds made up of different one-dimensional(1D)fibers in tissue engineering.In particular,we focus on the improvements made in recent years,especially in the fields of wound healing,angiogenesis,and tissue regeneration.
基金supported by National Key R&D Program of China (2021YFA0805100)National Natural Science Foundation of China (92268202,82270485,81830013,81970363)+7 种基金the Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences (2023-PT320-03)Guangdong Basic and Applied Basic Research FoundationChina (2019B1515120092)the Science and Technology Planning Project of GuangzhouChina (202103000016)the Science and Technology Planning Project of Guangdong Province (2023B1212060018)the Sun Yat-sen University Clinical Research 5010 ProgramProgram of National Key Clinical Specialties。
文摘Diabetic foot ulcers(DFUs)are a serious vascular disease.Currently,no effective methods are available for treating DFUs.Pro-protein convertase subtilisin/kexin type 9(PCSK9)regulates lipid levels to promote atherosclerosis.However,the role of PCSK9 in DFUs remains unclear.In this study,we found that the expression of PCSK9 in endothelial cells(ECs)increased significantly under high glucose(HG)stimulation and in diabetic plasma and vessels.Specifically,PCSK9 promotes the E3 ubiquitin-protein ligase NEDD4 binding to vascular endothelial growth factor receptor 2(VEGFR2),which led to the ubiquitination of VEGFR2,resulting in its degradation and downregulation in ECs.Furthermore,PCSK9 suppresses the expression and activation of AKT,endothelial nitric oxide synthase(e NOS),and ERK1/2,leading to decreased nitric oxide(NO)production and increased superoxide anion(O_(2)^(·-))generation,which impairs vascular endothelial function and angiogenesis.Importantly,using evolocumab to limit the increase in PCSK9 expression blocked the HG-induced inhibition of NO production and the increase in O_(2)^(·-)production,as well as inhibited the phosphorylation and expression of AKT,e NOS,and ERK1/2.Moreover,evolocumab improved vascular endothelial function and angiogenesis,and promoted wound healing in diabetes.Our findings suggest that targeting PCSK9 is a novel therapeutic approach for treating DFUs.
基金supported by the National Key Research and Development Program of China(2021YFA0805100)the National Natural Science Foundation of China(81830013,81770241,81970363,82000362,92268202,81170271,81370370,81490531,81670392,81600382)+6 种基金the National Natural Science Foundation of China Distinguished Young Scholar Grant(81325001)“973 Project”from the Ministry of Science and Technology of China(2009CB522104)Guangdong Basic and Applied Basic Research Foundation,China(2019B1515120092)the Science and Technology Planning Project of Guangzhou,China(202103000016)the Changjiang Scholars Program from the Ministry of Education of Chinathe Sun Yat-sen University Clinical Research 5010 Programthe Program of National Key Clinical Specialties。
文摘Cardiopulmonary bypass has been speculated to elicit systemic inflammation to initiate acute lung injury(ALI), including acute respiratory distress syndrome(ARDS), in patients after cardiac surgery. We previously found that post-operative patients showed an increase in endothelial cell-derived extracellular vesicles(eEVs) with components of coagulation and acute inflammatory responses. However, the mechanism underlying the onset of ALI owing to the release of e EVs after cardiopulmonary bypass, remains unclear. Plasma plasminogenactivated inhibitor-1(PAI-1) and eEV levels were measured in patients with cardiopulmonary bypass. Endothelial cells and mice(C57BL/6,Toll-like receptor 4 knockout(TLR4^(-/-))) and inducible nitric oxide synthase knockout(iNOS^(-/-)) were challenged with eEVs isolated from PAI-1-stimulated endothelial cells. Plasma PAI-1 and eEVs were remarkably enhanced after cardiopulmonary bypass. Plasma PAI-1 elevation was positively correlated with the increase in eEVs. The increase in plasma PAI-1 and eEV levels was associated with post-operative ARDS. The eEVs derived from PAI-1-stimulated endothelial cells could recognize TLR4 to stimulate a downstream signaling cascade identified as the Janus kinase 2/3(JAK2/3)-signal transducer and activator of transcription 3(STAT3)-interferon regulatory factor 1(IRF-1)pathway, along with i NOS induction, and cytokine/chemokine production in vascular endothelial cells and C57BL/6 mice, ultimately contributing to ALI. ALI could be attenuated by JAK2/3 or STAT3 inhibitors(AG490 or S3I-201, respectively), and was relieved in TLR4-/-and iNOS-/-mice. eEVs activate the TLR4/JAK3/STAT3/IRF-1 signaling pathway to induce ALI/ARDS by delivering follistatin-like protein 1(FSTL1), and FSTL1 knockdown in eEVs alleviates eEV-induced ALI/ARDS. Our data thus demonstrate that cardiopulmonary bypass may increase plasma PAI-1 levels to induce FSTL1-enriched eEVs, which target the TLR4-mediated JAK2/3/STAT3/IRF-1 signaling cascade and form a positive feedback loop, leading to ALI/ARDS after cardiac surgery. Our findings provide new insight into the molecular mechanisms and therapeutic targets for ALI/ARDS after cardiac surgery.
基金We thank members of the Huang laboratory for advice and support.This work is supported by National Key R&D Program of China(2021YFA1302500)National Natural Science Foundation of China(81873463 to Z.-P.H.and 82000385 to X.H.)+1 种基金Guangdong Basic and Applied Basic Research Foundation(2019B151502003 to Z.-P.H.)the donation for scientific research from the Terry Fox Foundation to Z.-P.H.
文摘Chronic heart failure is the end stage of cardiac diseases.With a high prevalence and a high mortality rate worldwide,chronic heart failure is one of the heaviest health-related burdens.In addition to the standard neurohormonal blockade therapy,several medications have been developed for chronic heart failure treatment,but the population-wide improvement in chronic heart failure prognosis over time has been modest,and novel therapies are still needed.
基金We sincerely acknowledge the funding from the Science and Technology Planning Project of Shenzhen Municipality(Nos.JCYJ20180306174831458 and JCYJ20190807155801657)the National Natural Science Foundation of China(Nos.51973243 and 52173150)+3 种基金Guangdong Innovative and Entrepreneurial Research Team Program(No.2016ZT06S029)Key international(regional)cooperative research projects of the National Natural Science Foundation of China(No.5181001045)The International Science and technology cooperation project of Shenzhen Municipality(No.GJHZ20200731095008026)Public welfare project of Futian District Health Bureau(No.FTWS2021001).
文摘Wound ulceration caused by diabetes is a typical chronic wound wherein healing the local tissue is difficult due to lack of blood vessels and tissue necrosis caused by the long-term accumulation of free radicals.Near-field electrospinning(NFES)is an innovative technology used to produce micro-nano-scaled,controllable sequencing fibers.In this study,we constructed a novel wound dressing based on the NFES polycaprolactone(PCL)fiber network and modified gelatin with methacrylic anhydride(GelMA)hydrogel to promote angiogenesis and the re-epithelialization of diabetic wounds.An angiogenic and antioxidant drug named deferoxamine(DFO)was encapsulated in a GelMA hydrogel to achieve a slow-release effect that is more suitable for chronic wounds.The cell adhesion experiment showed that the cells could attach to the fibers in the dressing group having a network of PCL fibers on the surface and grow along the direction of the fibers,which in turn,effectively regulates cell behavior from the physical structure.Additionally,the large pore size(~500μm)of the network allowed the cells to penetrate the pores and enter the surface of the hydrogel without being blocked out.Besides,the composite dressing had a notable effect on angiogenesis.Furthermore,antioxidation experiments confirmed that the DFO-loaded hydrogel exhibited antioxidant activity.Experimental animal models of diabetes showed that rats treated with the PCL-GelMA-DFO(PGD)hydrogel had faster ability of hemostasis,scab formation,and wound healing.In conclusion,the PGD hydrogel effectively promoted the repair of chronic wounds.
基金This research was financially supported by the National Key R&D Program of China 2021YFA0805100National Natural Science Foundation of China(Grant Nos.92268202,81830013,81970363,82270485,and 81770241)+3 种基金Guangdong Basic and Applied Basic Research Foundation(Grant 2019B1515120092)Science and Technology Planning Project of Guangzhou,China 202103000016Sun Yat-sen University Clinical Research 5010 Program(2014002)the Program of National Key Clinical Specialties.Figures 7a,8d and Supplementary Fig.S14a were created with BioRender.com.
文摘Normal high-density lipoprotein(nHDL)can induce angiogenesis in healthy individuals.However,HDL from patients with coronary artery disease undergoes various modifications,becomes dysfunctional(dHDL),and loses its ability to promote angiogenesis.Here,we identified a long non-coding RNA,HDRACA,that is involved in the regulation of angiogenesis by HDL.In this study,we showed that nHDL downregulates the expression of HDRACA in endothelial cells by activating WW domain-containing E3 ubiquitin protein ligase 2,which catalyzes the ubiquitination and subsequent degradation of its transcription factor,Kruppel-like factor 5,via sphingosine 1-phosphate(S1P)receptor 1.In contrast,dHDL with lower levels of S1P than nHDL were much less effective in decreasing the expression of HDRACA.HDRACA was able to bind to Ras-interacting protein 1(RAIN)to hinder the interaction between RAIN and vigilin,which led to an increase in the binding between the vigilin protein and proliferating cell nuclear antigen(PCNA)mRNA,resulting in a decrease in the expression of PCNA and inhibition of angiogenesis.The expression of human HDRACA in a hindlimb ischemia mouse model inhibited the recovery of angiogenesis.Taken together,these findings suggest that HDRACA is involved in the HDL regulation of angiogenesis,which nHDL inhibits the expression of HDRACA to induce angiogenesis,and that dHDL is much less effective in inhibiting HDRACA expression,which provides an explanation for the decreased ability of dHDL to stimulate angiogenesis.
基金This study is supported by the State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center,Sun Yat-sen University,and Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science,Guangzhou 510060,P.R.Chinathe National Natural Science Foundation of China(82150710555 and 82220108016 to X.Li,81970823 to Jin Yao and 81830013 to J.O.)+4 种基金a Key Research and Development Plan of Shandong Province(2016GSF201100)the Fundamental Research Funds for the Central Universities(19ykpy151)the long-term structural Methusalem funding by the Flemish Government,Belgiumthe Deutsche Forschungsge-meinschaft(Project No.:394046768-SFB1366)the DZHK partner site Mannheim/Heidelberg to H.F.L.,an ERA PerMed 2020 JTC grant“PROGRESS”.
文摘Although VEGF-B was discovered as a VEGF-A homolog a long time ago,the angiogenic effect of VEGF-B remains poorly understood with limited and diverse findings from different groups.Notwithstanding,drugs that inhibit VEGF-B together with other VEGF family members are being used to treat patients with various neovascular diseases.It is therefore critical to have a better understanding of the angiogenic effect of VEGF-B and the underlying mechanisms.Using comprehensive in vitro and in vivo methods and models,we reveal here for the first time an unexpected and surprising function of VEGF-B as an endogenous inhibitor of angiogenesis by inhibiting the FGF2/FGFR1 pathway when the latter is abundantly expressed.Mechanistically,we unveil that VEGF-B binds to FGFR1,induces FGFR1/VEGFR1 complex formation,and suppresses FGF2-induced Erk activation,and inhibits FGF2-driven angiogenesis and tumor growth.Our work uncovers a previously unrecognized novel function of VEGF-B in tethering the FGF2/FGFR1 pathway.Given the anti-angiogenic nature of VEGF-B under conditions of high FGF2/FGFR1 levels,caution is warranted when modulating VEGF-B activity to treat neovascular diseases.
基金supported by the National Natural Science Foundation of China(81830013,82100424,92268202,81970363)the National Key Research and Development Program of China(2021YFA0805100)+4 种基金Guangdong Basic and Applied Basic Research Foundation(2019B1515120092)Science and Technology Planning Project of GuangzhouChina(202103000016)the Sun Yat-sen University Clinical Research 5010 Program(2014002)Program of National Key Clinical Specialties。
文摘We previously demonstrated that normal high-density lipoprotein(nHDL)can promote angiogenesis,whereas HDL from patients with coronary artery disease(d HDL)is dysfunctional and impairs angiogenesis.Autophagy plays a critical role in angiogenesis,and HDL regulates autophagy.However,it is unclear whether n HDL and d HDL regulate angiogenesis by affecting autophagy.Endothelial cells(ECs)were treated with n HDL and d HDL with or without an autophagy inhibitor.Autophagy,endothelial nitric oxide synthase(e NOS)expression,miRNA expression,nitric oxide(NO)production,superoxide anion(O2^(·-))generation,EC migration,and tube formation were evaluated.n HDL suppressed the expression of miR-181a-5p,which promotes autophagy and the expression of e NOS,resulting in NO production and the inhibition of O2^(·-)generation,and ultimately increasing in EC migration and tube formation.d HDL showed opposite effects compared to n HDL and ultimately inhibited EC migration and tube formation.We found that autophagy-related protein 5(ATG5)was a direct target of miR-181a-5p.ATG5 silencing or miR-181a-5p mimic inhibited n HDL-induced autophagy,e NOS expression,NO production,EC migration,tube formation,and enhanced O2^(·-)generation,whereas overexpression of ATG5 or miR-181a-5p inhibitor reversed the above effects of d HDL.ATG5 expression and angiogenesis were decreased in the ischemic lower limbs of hypercholesterolemic low-density lipoprotein receptor null(LDLr^(-/-))mice when compared to C57BL/6 mice.ATG5 overexpression improved angiogenesis in ischemic hypercholesterolemic LDLr^(-/-)mice.Taken together,nHDL was able to stimulate autophagy by suppressing miR-181a-5p,subsequently increasing e NOS expression,which generated NO and promoted angiogenesis.In contrast,d HDL inhibited angiogenesis,at least partially,by increasing miR-181a-5p expression,which decreased autophagy and e NOS expression,resulting in a decrease in NO production and an increase in O2^(·-)generation.Our findings reveal a novel mechanism by which HDL affects angiogenesis by regulating autophagy and provide a therapeutic target for d HDL-impaired angiogenesis.
基金supported by the National Natural Science Foundation of China(92057113 and 32122027)the Natural Science Foundation of Guangdong Province(2021A1515012489 and 2022A1515011819)+1 种基金the National Natural Science Foundation of China(82200280)the China Postdoctoral Science Foundation(2022M713573).
文摘Cardiac fibrosis is a pathological response characterized by excessive deposition of fibrous connective tissue within the heart.It typically occurs following cardiac injuries or diseases.However,the lack of suitable models for disease modeling and high-throughput drug discovery has hindered the establishment of an effective treatments for cardiac fibrosis.The emergence and rapid progress of stem-cell and lineage reprogramming technology offer an unprecedented opportunity to develop an improved humanized and patient-specific model for studying cardiac fibrosis,providing a platform for screening potential drugs and synchronously elucidating the underlying molecular mechanisms.Furthermore,reprogramming cardiac fibroblasts into cardiomyocyte-like cells to reduce scar volume and induce myocardial tissue regeneration is a promising approach in treating cardiac fibrosis.In this review,we summarize the current advancements in stem cell technologies applied to study cardiac fibrosis and provide insights for future investigations into its mechanisms,drug discovery as well as therapy method.
文摘Background and Aims:Increasing utilization of extended criteria donor leads to an increasing rate of early allograft failure after liver transplantation.However,consensus of definition of early allograft failure is lacking.Methods:A retrospective,multicenter study was performed to validate the Liver Graft Assessment Following Transplantation(L-GrAFT)risk model in a Chinese cohort of 942 adult patients undergoing primary liver transplantation at three Chinese centers.L-GrAFT(L-GrAFT7 and L-GrAFT10)was compared with existing models:the Early Allograft Failure Simplified Estimation(EASE)score,the model of early allograft function(MEAF),and the Early Allograft Dysfunction(EAD)model.Univariate and multivariate logistic regression were used to find risk factors of L-GrAFT high-risk group.Results:L-GrAFT7 had an area under the curve of 0.85 in predicting 90-day graft survival,significantly superior to MEAF[area under the curve(AUC=0.78,p=0.044)]and EAD(AUC=0.78,p=0.006),while there was no statistical significance between the predicting abilities of L-GrAFT7 and EASE(AUC=0.84,p>0.05).Furthermore,L-GrAFT7 maintains good predicting ability in the subgroup of high-donor risk index(DRI)cases(AUC=0.83 vs.MEAF,p=0.007 vs.EAD,p=0.014)and recipients of donors after cardiac death(AUC=0.92 vs.EAD,p<0.001).Through multivariate analysis,pretransplant bilirubin level,units of packed red blood cells,and the DRI score were selected as independent risk factors of a L-GrAFT7 high-risk group.Conclusions:The accuracy of L-GrAFT7 in predicting early allograft failure was validated in a Chinese multicenter cohort,indicating that it has the potential to become an accurate endpoint of clinical practice and transitional study of machine perfusion.
