Distributed storage can store data in multiple devices or servers to improve data security.However,in today’s explosive growth of network data,traditional distributed storage scheme is faced with some severe challeng...Distributed storage can store data in multiple devices or servers to improve data security.However,in today’s explosive growth of network data,traditional distributed storage scheme is faced with some severe challenges such as insufficient performance,data tampering,and data lose.A distributed storage scheme based on blockchain has been proposed to improve security and efficiency of traditional distributed storage.Under this scheme,the following improvements have been made in this paper.This paper first analyzes the problems faced by distributed storage.Then proposed to build a new distributed storage blockchain scheme with sharding blockchain.The proposed scheme realizes the partitioning of the network and nodes by means of blockchain sharding technology,which can improve the efficiency of data verification between nodes.In addition,this paper uses polynomial commitment to construct a new verifiable secret share scheme called PolyVSS.This new scheme is one of the foundations for building our improved distributed storage blockchain scheme.Compared with the previous scheme,our new scheme does not require a trusted third party and has some new features such as homomorphic and batch opening.The security of VSS can be further improved.Experimental comparisons show that the proposed scheme significantly reduces storage and communication costs.展开更多
G protein-coupled receptor kinase 2(GRK2)participates in the phosphorylation and desensitization of G protein-coupled receptor(GPCR),impacting various biological processes such as inflammation and cell proliferation.D...G protein-coupled receptor kinase 2(GRK2)participates in the phosphorylation and desensitization of G protein-coupled receptor(GPCR),impacting various biological processes such as inflammation and cell proliferation.Dysregulated expression and activity of GRK2 have been reported in multiple cells in rheumatoid arthritis(RA).However,whether and how GRK2 regulates synovial hyperplasia and fibroblast-like synoviocytes(FLSs)proliferation is poorly understood.In this study,we investigated the regulation of GRK2 and its biological function in RA.We found that GRK2 transmembrane activity was increased in FLSs of RA patients and collagen-induced arthritis(CIA)rats.Additionally,we noted a positive correlation between high GRK2 expression on the cell membrane and serological markers associated with RA and CIA.Immunoprecipitationemass spectrometry and pulldown analyses revealed tumor necrosis factor receptor-associated factor 2(TRAF2)as a novel substrate of GRK2.Furthermore,surface plasmon resonance(SPR)and molecular docking assays determined that the C-terminus of GRK2 binds to the C-terminus of TRAF2 at the Gln340 residue.GRK2 knockdown and the GRK2 inhibitor CP-25 attenuated synovial hyperplasia and FLS proliferation in CIA both in vitro and in vivo by decreasing GRK2 membrane expression and activity.Mechanistically,increased GRK2 transmembrane activity contributed to the recruitment of TRAF2 on the cell membrane,promoting GRK2-RAF2 interactions that facilitate the recruitment of the E3 ubiquitin ligase TRIM47 to TRAF2.This enhanced TRAF2 Lys63 polyubiquitylation and induced nuclear factor(NF)-kB activation,leading to synovial hyperplasia and abnormal proliferation of FLSs.Our study provides a mechanistic and preclinical rationale for further evaluation of GRK2 as a therapeutic target for RA.展开更多
Rheumatoid arthritis(RA)is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes(FLS).The up-regulated cellular membrane expression of G protein coupled receptor k...Rheumatoid arthritis(RA)is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes(FLS).The up-regulated cellular membrane expression of G protein coupled receptor kinase 2(GRK2)of FLS plays a critical role in RA progression,the increase of GRK2 translocation activity promotes dysfunctional prostaglandin E4 receptor(EP4)signaling and FLS abnormal proliferation.Recently,although our group found that paeoniflorin-6’-O-benzene sulfonate(CP-25),a novel compound,could reverse FLS dysfunction via GRK2,little is known as to how GRK2 translocation activity is suppressed.Our findings revealed that GRK2 expression up-regulated and EP4 expression down-regulated in synovial tissues of RA patients and collagen-induced arthritis(CIA)rats,and prostaglandin E2(PGE2)level increased in arthritis.CP-25 could down-regulate GRK2 expression,up-regulate EP4 expression,and improve synovitis of CIA rats.CP-25 and GRK2 inhibitors(paroxetine or GSK180736 A)inhibited the abnormal proliferation of FLS in RA patients and CIA rats by down-regulating GRK2 translocation to EP4 receptor.The results of microscale thermophoresis(MST),cellular thermal shift assay,and inhibition of kinase activity assay indicated that CP-25 could directly target GRK2,increase the protein stability of GRK2 in cells,and inhibit GRK2 kinase activity.The docking of CP-25 and GRK2 suggested that the kinase domain of GRK2 might be an important active pocket for CP-25.G201,K220,K230,A321,and D335 in kinase domain of GRK2 might form hydrogen bonds with CP-25.Site-directed mutagenesis and co-immunoprecipitation assay further revealed that CP-25 down-regulated the interaction of GRK2 and EP4 via controlling the key amino acid residue of Ala321 of GRK2.Our data demonstrate that FLS proliferation is regulated by GRK2 translocation to EP4.Targeted inhibition of GRK2 kinase domain by CP-25 improves FLS function and represents an innovative drug for the treatment of RA by targeting GRK2.展开更多
基金This work was supported by the National Natural Science Foundation of China under Grant 62072249,61772280,61772454,62072056.J.Wang and Y.Ren received the grants,and the URL of the sponsors’website is http://www.nsfc.gov.cn/This work was also supported by the Project of Transformation and Upgrading of Industries and Information Technologies of Jiangsu Province(No.JITC-1900AX2038/01).X.Yu received the grant,and the URL of the sponsors’website is http://gxt.jiangsu.gov.cn/.
文摘Distributed storage can store data in multiple devices or servers to improve data security.However,in today’s explosive growth of network data,traditional distributed storage scheme is faced with some severe challenges such as insufficient performance,data tampering,and data lose.A distributed storage scheme based on blockchain has been proposed to improve security and efficiency of traditional distributed storage.Under this scheme,the following improvements have been made in this paper.This paper first analyzes the problems faced by distributed storage.Then proposed to build a new distributed storage blockchain scheme with sharding blockchain.The proposed scheme realizes the partitioning of the network and nodes by means of blockchain sharding technology,which can improve the efficiency of data verification between nodes.In addition,this paper uses polynomial commitment to construct a new verifiable secret share scheme called PolyVSS.This new scheme is one of the foundations for building our improved distributed storage blockchain scheme.Compared with the previous scheme,our new scheme does not require a trusted third party and has some new features such as homomorphic and batch opening.The security of VSS can be further improved.Experimental comparisons show that the proposed scheme significantly reduces storage and communication costs.
基金supported by National Natural Science Foundation of China(Nos.82104187,82173824,and 82430114)Natural Science Foundation of Anhui Province(No.2108085QH382,China)+2 种基金Project funded by China Postdoctoral Science Foundation(Nos.2021T140002 and 2021M700184)the Postdoctoral Science Foundation of Anhui Province(No.2020B430,China)Postgraduate Innovation Research and Practice Program of Anhui Medical University(No.YJS20230162,China).
文摘G protein-coupled receptor kinase 2(GRK2)participates in the phosphorylation and desensitization of G protein-coupled receptor(GPCR),impacting various biological processes such as inflammation and cell proliferation.Dysregulated expression and activity of GRK2 have been reported in multiple cells in rheumatoid arthritis(RA).However,whether and how GRK2 regulates synovial hyperplasia and fibroblast-like synoviocytes(FLSs)proliferation is poorly understood.In this study,we investigated the regulation of GRK2 and its biological function in RA.We found that GRK2 transmembrane activity was increased in FLSs of RA patients and collagen-induced arthritis(CIA)rats.Additionally,we noted a positive correlation between high GRK2 expression on the cell membrane and serological markers associated with RA and CIA.Immunoprecipitationemass spectrometry and pulldown analyses revealed tumor necrosis factor receptor-associated factor 2(TRAF2)as a novel substrate of GRK2.Furthermore,surface plasmon resonance(SPR)and molecular docking assays determined that the C-terminus of GRK2 binds to the C-terminus of TRAF2 at the Gln340 residue.GRK2 knockdown and the GRK2 inhibitor CP-25 attenuated synovial hyperplasia and FLS proliferation in CIA both in vitro and in vivo by decreasing GRK2 membrane expression and activity.Mechanistically,increased GRK2 transmembrane activity contributed to the recruitment of TRAF2 on the cell membrane,promoting GRK2-RAF2 interactions that facilitate the recruitment of the E3 ubiquitin ligase TRIM47 to TRAF2.This enhanced TRAF2 Lys63 polyubiquitylation and induced nuclear factor(NF)-kB activation,leading to synovial hyperplasia and abnormal proliferation of FLSs.Our study provides a mechanistic and preclinical rationale for further evaluation of GRK2 as a therapeutic target for RA.
基金supported by the Key Project of National Natural Science Foundation of China(No.81330081)Surface Project of National Natural Science Foundation of China(No.81673444)Youth Science Fund Project of National Natural Science Foundation of China(No.81502123)
文摘Rheumatoid arthritis(RA)is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes(FLS).The up-regulated cellular membrane expression of G protein coupled receptor kinase 2(GRK2)of FLS plays a critical role in RA progression,the increase of GRK2 translocation activity promotes dysfunctional prostaglandin E4 receptor(EP4)signaling and FLS abnormal proliferation.Recently,although our group found that paeoniflorin-6’-O-benzene sulfonate(CP-25),a novel compound,could reverse FLS dysfunction via GRK2,little is known as to how GRK2 translocation activity is suppressed.Our findings revealed that GRK2 expression up-regulated and EP4 expression down-regulated in synovial tissues of RA patients and collagen-induced arthritis(CIA)rats,and prostaglandin E2(PGE2)level increased in arthritis.CP-25 could down-regulate GRK2 expression,up-regulate EP4 expression,and improve synovitis of CIA rats.CP-25 and GRK2 inhibitors(paroxetine or GSK180736 A)inhibited the abnormal proliferation of FLS in RA patients and CIA rats by down-regulating GRK2 translocation to EP4 receptor.The results of microscale thermophoresis(MST),cellular thermal shift assay,and inhibition of kinase activity assay indicated that CP-25 could directly target GRK2,increase the protein stability of GRK2 in cells,and inhibit GRK2 kinase activity.The docking of CP-25 and GRK2 suggested that the kinase domain of GRK2 might be an important active pocket for CP-25.G201,K220,K230,A321,and D335 in kinase domain of GRK2 might form hydrogen bonds with CP-25.Site-directed mutagenesis and co-immunoprecipitation assay further revealed that CP-25 down-regulated the interaction of GRK2 and EP4 via controlling the key amino acid residue of Ala321 of GRK2.Our data demonstrate that FLS proliferation is regulated by GRK2 translocation to EP4.Targeted inhibition of GRK2 kinase domain by CP-25 improves FLS function and represents an innovative drug for the treatment of RA by targeting GRK2.
