Exosomes(EXOs)play an important role in the progression of breast cancer.EXOs,with a diameter of approximately 100 nm,have a simple structure but diverse functions,and can affect the development of breast cancer throu...Exosomes(EXOs)play an important role in the progression of breast cancer.EXOs,with a diameter of approximately 100 nm,have a simple structure but diverse functions,and can affect the development of breast cancer through signal transduction and molecular transfer,etc.Angiogenesis provides nutrients for the growth and metastasis of breast cancer and is a crucial part of tumor progression.The mechanism of tumor angiogenesis is complex.The VEGF/VEGFR pathway promotes angiogenesis by regulating the activities of ECs.Hypoxia,a common feature in the tumor microenvironment,as a key regulator,can affect angiogenesis in multiple aspects such as the transfer of miRNAs in EXOs,protein transport,extracellular matrix regulation,and metabolic adaptation.The Notch pathway has a bidirectional regulatory role in breast cancer angiogenesis,and different molecules can promote or inhibit angiogenesis.EXOs secreted by breast cancer cells are rich in angiogenic factors.Components such as proteins and nucleic acids in EXOs can affect the functions and behaviors of vascular ECs,thereby influencing breast cancer angiogenesis.Research on the mechanisms of EXOs in breast cancer angiogenesis is of great significance for tumor treatment.EXOs are expected to become biomarkers for breast cancer diagnosis/prognosis.This research provides potential targets for in-depth understanding of the biological characteristics of breast cancer and the development of new treatment strategies.展开更多
Objective:To use the gene chip of pseudomonas aeruginosa as a research sample and to explore it at an omics level,aiming at elucidating the co-expression network characteristics of the virulence genes exoS and exoU of...Objective:To use the gene chip of pseudomonas aeruginosa as a research sample and to explore it at an omics level,aiming at elucidating the co-expression network characteristics of the virulence genes exoS and exoU of pseudomonas aeruginosa in the lower respiratory tract from the perspective of molecular biology and identifying its key regulatory genes.Methods:From March 2016 to May 2018,312 patients infected with pseudomonas aeruginosa in the lower respiratory tract who were admitted to Department of Respiratory Medicine of Baogang Hospital and given follow-up treatments in the hospital were selected as subjects by use of cluster sampling.Alveolar lavage fluid and sputum collected from those patients were used as biological specimens.The genes of pseudomonas aeruginosa were detected with the help of oligonucleotide probes to make a pre-processing of chip data.A total of 8 common antibiotics(ceftazidime,gentamicin,piperacillin,amikacin,ciprofloxacin,levofloxacin,doripenem and ticarcillin)against Gram-negative bacteria were selected to determine the drug resistance of biological specimens.MCODE algorithm was used to construct a co-expression network model of the drug-resistance genes focused on exoS/exoU.Results:The expression level of exoS/exoU in the drug-resistance group was significantly higher than that in the non-resistance group(p<0.05).The top 5 differentially expressed genes in the alveolar lavage fluid specimens from the drug-resistance group were RAC1,ITGB1,ITGB5,CRK and IGF1R in the order from high to low.In the sputum specimens,the top 5 differentially expressed genes were RAC1,CRK,IGF1R,ITGB1 and ITGB5.In the alveolar lavage fluid specimens,only RAC1 had a positive correlation with the expression of exoS and exoU(p<0.05).In the sputum specimens,RAC1,ITGB1,ITGB5,CRK and IGF1R were positively correlated with the expression of exoS and exoU(p<0.05).The genes included in the co-expression network contained exoS,exoU,RAC1,ITGB1,ITGB5,CRK,CAMK2D,RHOA,FLNA,IGF1R,TGFBR2 and FOS.Among them,RAC1 had a highest score in the aspect of regulatory ability(72.00)and the largest number of regulatory genes(6);followed by ITGB1,ITGB5 and CRK genes.Conclusions:The high expression of exoS and exoU in the sputum specimens suggests that pseudomonas aeruginosa has a higher probability to get resistant to antibiotics;RAC1,ITGB1,ITGB5 and CRK genes may be the key genes that can regulate the expression of exoS and exoU.展开更多
Background Neuroinflammation participates in the pathogenesis of subarachnoid haemorrhage(SAH);however,no effective treatments exist.MicroRNAs regulate several aspects of neuronal dysfunction.In a previous study,we fo...Background Neuroinflammation participates in the pathogenesis of subarachnoid haemorrhage(SAH);however,no effective treatments exist.MicroRNAs regulate several aspects of neuronal dysfunction.In a previous study,we found that exosomal miR-486-3p is involved in the pathophysiology of SAH.Targeted delivery of miR-486-3p without blood-brain barrier(BBB)restriction to alleviate SAH is a promising neuroinflammation approach.Methods In this study,we modified exosomes(Exo)to form an RVG-miR 486-3p Exo(Exo/miR)to achieve targeted delivery of miR-486-3p to the brain.Neurological scores,brain water content,BBB damage,flow cytometry and FJC staining were used to determine the effect of miR-486-3p on SAH.Western blot analysis,ELISA and RT-qPCR were used to measure relevant protein and mRNA levels.Immunofluorescence staining and laser confocal detection were used to measure the expression of mitochondria,lysosomes and autophagosomes,and transmission electron microscopy was used to observe the level of mitophagy in the brain tissue of mice after SAH.Results Tail vein injection of Exo/miR improved targeting of miR-486-3p to the brains of SAH mice.The injection reduced levels of neuroinflammation-related factors by changing the phenotype switching of microglia,inhibiting the expression of sirtuin 2(SIRT2)and enhancing mitophagy.miR-486-3p treatment alleviated neurobehavioral disorders,brain oedema,BBB damage and neurodegeneration.Further research found that the mechanism was achieved by regulating the acetylation level of peroxisome proliferator-activated receptorγcoactivator l alpha(PGC-1α)after SIRT2 enters the nucleus.Conclusion Exo/miR treatment attenuates neuroinflammation after SAH by inhibiting SIRT2 expression and stimulating mitophagy,suggesting potential clinical applications.展开更多
The management of diabetic wounds remains a critical therapeutic challenge. Platelet-rich plasma (PRP) gel, PRP-derived exosomes (PRP-Exos), and mesenchymal stem cell-derived exosomes (MSC-Exos) have demonstrated ther...The management of diabetic wounds remains a critical therapeutic challenge. Platelet-rich plasma (PRP) gel, PRP-derived exosomes (PRP-Exos), and mesenchymal stem cell-derived exosomes (MSC-Exos) have demonstrated therapeutic potential in wound treatment. Unfortunately, their poor mechanical properties, the short half-lives of growth factors (GFs), and the burst release of GFs and exosomes have limited their clinical applications. Furthermore, proteases in diabetic wounds degrade GFs, which hampers wound repair. Silk fibroin is an enzyme-immobilization biomaterial that could protect GFs from proteases. Herein, we developed novel dual-crosslinked hydrogels based on silk protein (SP) (sericin and fibroin), including SP@PRP, SP@MSC-Exos, and SP@PRP-Exos, to promote diabetic wound healing synergistically. SP@PRP was prepared from PRP and SP using calcium gluconate/thrombin as agonist, while SP@PRP-Exos and SP@MSC-Exos were derived from exosomes and SP with genipin as crosslinker. SP provided improved mechanical properties and enabled the sustained release of GFs and exosomes, thereby overcoming the limitations of PRP and exosomes in wound healing. The dual-crosslinked hydrogels displayed shear-induced thinning, self-healing, and eradication of microbial biofilms in a bone-mimicking environment. In vivo, the dual-crosslinked hydrogels contributed to faster diabetic wound healing than PRP and SP by upregulating GFs expression, down-regulating matrix metalloproteinase-9 expression, and by promoting an anti-NETotic effect, angiogenesis, and re-epithelialization. Hence, these dual-crosslinked hydrogels have the potential to be translated into a new generation of diabetic wound dressings.展开更多
Pseudomonas aeruginosa is a significant pathogen mainly causing healthcare-associated infections(HAIs).Newly emerging high-risk clones of P.aeruginosa with elevated virulence profiles furtherly cause severe community-...Pseudomonas aeruginosa is a significant pathogen mainly causing healthcare-associated infections(HAIs).Newly emerging high-risk clones of P.aeruginosa with elevated virulence profiles furtherly cause severe community-acquired infections(CAIs).Usually,it is not common for P.aeruginosa to co-carry exoU and exoS genes,encoding two type III secretion system(T3SS)effectors.The pathogenicity mechanism of exoS+/exoU+strains of P.aeruginosa remains unclear.Here,we provide detailed evidence for a subset of hypervirulent P.aeruginosa strains,which abundantly co-express and secrete the T3SS effectors ExoS and ExoU.The exoS+/exoU+P.aeruginosa strains were available to cause both HAIs and CAIs.The CAI-associated strains could elicit severe inflammation and hemorrhage,leading to higher death rates in a murine acute pneumonia model,and had great virulence potential in establishing chronic infections,demonstrating hypervirulence when compared to PAO1(exoS+/exoU-)and PA14(exoS-/exoU+).Both ExoS and ExoU were co-expressed and co-secreted in abundance in exoS+/exoU+strains.Their abundant protein secretion could boost exoS+/exoU+strains’potentials for cytotoxicity in vitro and pathogenicity in vivo.Genomic evidence indicates that exoU acquisition is likely mediated by horizontal gene transfer(HGT)of the pathogenicity island PAPI-2,while deletion of exoU was sufficient to mitigate virulence in the exoS+/exoU+strains.Furthermore,bioinformatics analysis showed that such exoS+/exoU+P.aeruginosa strains turned out to be widely distributed across the globe.Overall,the research provide detailed evidence for the high virulence and epidemicity of exoS+/exoU+strains of P.aeruginosa,highlighting an urgent need for surveillance against these high-risk hypervirulent strains.展开更多
文摘Exosomes(EXOs)play an important role in the progression of breast cancer.EXOs,with a diameter of approximately 100 nm,have a simple structure but diverse functions,and can affect the development of breast cancer through signal transduction and molecular transfer,etc.Angiogenesis provides nutrients for the growth and metastasis of breast cancer and is a crucial part of tumor progression.The mechanism of tumor angiogenesis is complex.The VEGF/VEGFR pathway promotes angiogenesis by regulating the activities of ECs.Hypoxia,a common feature in the tumor microenvironment,as a key regulator,can affect angiogenesis in multiple aspects such as the transfer of miRNAs in EXOs,protein transport,extracellular matrix regulation,and metabolic adaptation.The Notch pathway has a bidirectional regulatory role in breast cancer angiogenesis,and different molecules can promote or inhibit angiogenesis.EXOs secreted by breast cancer cells are rich in angiogenic factors.Components such as proteins and nucleic acids in EXOs can affect the functions and behaviors of vascular ECs,thereby influencing breast cancer angiogenesis.Research on the mechanisms of EXOs in breast cancer angiogenesis is of great significance for tumor treatment.EXOs are expected to become biomarkers for breast cancer diagnosis/prognosis.This research provides potential targets for in-depth understanding of the biological characteristics of breast cancer and the development of new treatment strategies.
文摘Objective:To use the gene chip of pseudomonas aeruginosa as a research sample and to explore it at an omics level,aiming at elucidating the co-expression network characteristics of the virulence genes exoS and exoU of pseudomonas aeruginosa in the lower respiratory tract from the perspective of molecular biology and identifying its key regulatory genes.Methods:From March 2016 to May 2018,312 patients infected with pseudomonas aeruginosa in the lower respiratory tract who were admitted to Department of Respiratory Medicine of Baogang Hospital and given follow-up treatments in the hospital were selected as subjects by use of cluster sampling.Alveolar lavage fluid and sputum collected from those patients were used as biological specimens.The genes of pseudomonas aeruginosa were detected with the help of oligonucleotide probes to make a pre-processing of chip data.A total of 8 common antibiotics(ceftazidime,gentamicin,piperacillin,amikacin,ciprofloxacin,levofloxacin,doripenem and ticarcillin)against Gram-negative bacteria were selected to determine the drug resistance of biological specimens.MCODE algorithm was used to construct a co-expression network model of the drug-resistance genes focused on exoS/exoU.Results:The expression level of exoS/exoU in the drug-resistance group was significantly higher than that in the non-resistance group(p<0.05).The top 5 differentially expressed genes in the alveolar lavage fluid specimens from the drug-resistance group were RAC1,ITGB1,ITGB5,CRK and IGF1R in the order from high to low.In the sputum specimens,the top 5 differentially expressed genes were RAC1,CRK,IGF1R,ITGB1 and ITGB5.In the alveolar lavage fluid specimens,only RAC1 had a positive correlation with the expression of exoS and exoU(p<0.05).In the sputum specimens,RAC1,ITGB1,ITGB5,CRK and IGF1R were positively correlated with the expression of exoS and exoU(p<0.05).The genes included in the co-expression network contained exoS,exoU,RAC1,ITGB1,ITGB5,CRK,CAMK2D,RHOA,FLNA,IGF1R,TGFBR2 and FOS.Among them,RAC1 had a highest score in the aspect of regulatory ability(72.00)and the largest number of regulatory genes(6);followed by ITGB1,ITGB5 and CRK genes.Conclusions:The high expression of exoS and exoU in the sputum specimens suggests that pseudomonas aeruginosa has a higher probability to get resistant to antibiotics;RAC1,ITGB1,ITGB5 and CRK genes may be the key genes that can regulate the expression of exoS and exoU.
基金supported by grants from the National Natural Science Foundation of China(No.82271363(LY Wu),82301485(XX Chen),82201456(Y Zhou),82130037(CH.Hang))Natural Science Foundation of Jiangsu Province(No.2022JJ30060(Y Zhou))+1 种基金Anhui Provincial Department of Education Excellent Scientific Research and Innovation Team Project in Colleges and Universities(2022AH010073(NS Lai))Postgraduate Research&Practice Innovation Program of Jiangsu Province(SJCX24_0010(B Sheng)).
文摘Background Neuroinflammation participates in the pathogenesis of subarachnoid haemorrhage(SAH);however,no effective treatments exist.MicroRNAs regulate several aspects of neuronal dysfunction.In a previous study,we found that exosomal miR-486-3p is involved in the pathophysiology of SAH.Targeted delivery of miR-486-3p without blood-brain barrier(BBB)restriction to alleviate SAH is a promising neuroinflammation approach.Methods In this study,we modified exosomes(Exo)to form an RVG-miR 486-3p Exo(Exo/miR)to achieve targeted delivery of miR-486-3p to the brain.Neurological scores,brain water content,BBB damage,flow cytometry and FJC staining were used to determine the effect of miR-486-3p on SAH.Western blot analysis,ELISA and RT-qPCR were used to measure relevant protein and mRNA levels.Immunofluorescence staining and laser confocal detection were used to measure the expression of mitochondria,lysosomes and autophagosomes,and transmission electron microscopy was used to observe the level of mitophagy in the brain tissue of mice after SAH.Results Tail vein injection of Exo/miR improved targeting of miR-486-3p to the brains of SAH mice.The injection reduced levels of neuroinflammation-related factors by changing the phenotype switching of microglia,inhibiting the expression of sirtuin 2(SIRT2)and enhancing mitophagy.miR-486-3p treatment alleviated neurobehavioral disorders,brain oedema,BBB damage and neurodegeneration.Further research found that the mechanism was achieved by regulating the acetylation level of peroxisome proliferator-activated receptorγcoactivator l alpha(PGC-1α)after SIRT2 enters the nucleus.Conclusion Exo/miR treatment attenuates neuroinflammation after SAH by inhibiting SIRT2 expression and stimulating mitophagy,suggesting potential clinical applications.
基金supported by the National Natural Science Foundation of China(51973076)the Fundamental Research Funds for Central Universities(2020kfyXJJS035).
文摘The management of diabetic wounds remains a critical therapeutic challenge. Platelet-rich plasma (PRP) gel, PRP-derived exosomes (PRP-Exos), and mesenchymal stem cell-derived exosomes (MSC-Exos) have demonstrated therapeutic potential in wound treatment. Unfortunately, their poor mechanical properties, the short half-lives of growth factors (GFs), and the burst release of GFs and exosomes have limited their clinical applications. Furthermore, proteases in diabetic wounds degrade GFs, which hampers wound repair. Silk fibroin is an enzyme-immobilization biomaterial that could protect GFs from proteases. Herein, we developed novel dual-crosslinked hydrogels based on silk protein (SP) (sericin and fibroin), including SP@PRP, SP@MSC-Exos, and SP@PRP-Exos, to promote diabetic wound healing synergistically. SP@PRP was prepared from PRP and SP using calcium gluconate/thrombin as agonist, while SP@PRP-Exos and SP@MSC-Exos were derived from exosomes and SP with genipin as crosslinker. SP provided improved mechanical properties and enabled the sustained release of GFs and exosomes, thereby overcoming the limitations of PRP and exosomes in wound healing. The dual-crosslinked hydrogels displayed shear-induced thinning, self-healing, and eradication of microbial biofilms in a bone-mimicking environment. In vivo, the dual-crosslinked hydrogels contributed to faster diabetic wound healing than PRP and SP by upregulating GFs expression, down-regulating matrix metalloproteinase-9 expression, and by promoting an anti-NETotic effect, angiogenesis, and re-epithelialization. Hence, these dual-crosslinked hydrogels have the potential to be translated into a new generation of diabetic wound dressings.
基金supported by grants from the National Key R&D Program of China(2021YFC2302005)the Joint Funds of the International Development Research Center of Canada(109282-001)the National Key R&D Program of China(2021YFC2301004 and 2017YFE0125600).
文摘Pseudomonas aeruginosa is a significant pathogen mainly causing healthcare-associated infections(HAIs).Newly emerging high-risk clones of P.aeruginosa with elevated virulence profiles furtherly cause severe community-acquired infections(CAIs).Usually,it is not common for P.aeruginosa to co-carry exoU and exoS genes,encoding two type III secretion system(T3SS)effectors.The pathogenicity mechanism of exoS+/exoU+strains of P.aeruginosa remains unclear.Here,we provide detailed evidence for a subset of hypervirulent P.aeruginosa strains,which abundantly co-express and secrete the T3SS effectors ExoS and ExoU.The exoS+/exoU+P.aeruginosa strains were available to cause both HAIs and CAIs.The CAI-associated strains could elicit severe inflammation and hemorrhage,leading to higher death rates in a murine acute pneumonia model,and had great virulence potential in establishing chronic infections,demonstrating hypervirulence when compared to PAO1(exoS+/exoU-)and PA14(exoS-/exoU+).Both ExoS and ExoU were co-expressed and co-secreted in abundance in exoS+/exoU+strains.Their abundant protein secretion could boost exoS+/exoU+strains’potentials for cytotoxicity in vitro and pathogenicity in vivo.Genomic evidence indicates that exoU acquisition is likely mediated by horizontal gene transfer(HGT)of the pathogenicity island PAPI-2,while deletion of exoU was sufficient to mitigate virulence in the exoS+/exoU+strains.Furthermore,bioinformatics analysis showed that such exoS+/exoU+P.aeruginosa strains turned out to be widely distributed across the globe.Overall,the research provide detailed evidence for the high virulence and epidemicity of exoS+/exoU+strains of P.aeruginosa,highlighting an urgent need for surveillance against these high-risk hypervirulent strains.
