Receptors interaction protein 2(RIP2)is a specific adaptor molecule in the downstream of NOD2.The role of RIP2 during foot-and-mouth disease virus(FMDV)infection remains unknown.Here,our results showed that RIP2 inhib...Receptors interaction protein 2(RIP2)is a specific adaptor molecule in the downstream of NOD2.The role of RIP2 during foot-and-mouth disease virus(FMDV)infection remains unknown.Here,our results showed that RIP2 inhibited FMDV replication and played an important role in the activation of IFN-βand NF-κB signal pathways during FMDV infection.FMDV infection triggered RIP2 transcription,while it reduced the expression of RIP2 protein.Detailed analysis showed that FMDV 2B,2C,3C^(pro),and L^(pro) proteins were responsible for inducing the reduction of RIP2 protein.3C^(pro) and L^(pro) are viral proteinases that can induce the cleavage or reduction of many host proteins and block host protein synthesis.The carboxyl terminal 105-C114 and 135-C144 regions of 2B were essential for reduction of RIP2.Our results also showed that the N terminal 1-61 region of 2C were essential for the reduction of RIP2.The 2C-induced reduction of RIP2 was dependent on inducing the reduction of poly(A)-binding protein 1(PABPC1).The interaction between RIP2 and 2C was observed in the context of viral infection,and the residues 1-61 were required for the interaction.These data clarify novel mechanisms of reduction of RIP2 mediated by FMDV.展开更多
Background:Wound management of diabetic foot ulcers(DFUs)is a complex and challenging task,and existing strategies fail to meet clinical needs.Therefore,it is important to develop novel drug candidates and discover ne...Background:Wound management of diabetic foot ulcers(DFUs)is a complex and challenging task,and existing strategies fail to meet clinical needs.Therefore,it is important to develop novel drug candidates and discover new therapeutic targets.However,reports on peptides as molecular probes for resolving issues related to DFUs remain rare.This study utilized peptide RL-QN15 as an exogenous molecular probe to investigate the underlying mechanism of endogenous non-coding RNA in DFU wound healing.The aim was to generate novel insights for the clinical management of DFUs and identify potential drug targets.Methods:We investigated the wound-healing efficiency of peptide RL-QN15 under diabetic con-ditions using in vitro and in vivo experimental models.RNA sequencing,in vitro transfection,quantitative real-time polymerase chain reaction,western blotting,dual luciferase reporter gene detection,in vitro cell scratches,and cell proliferation and migration assays were performed to explore the potential mechanism underlying the promoting effects of RL-QN15 on DFU repair.Results:Peptide RL-QN15 enhanced the migration and proliferation of human immortalized keratinocytes(HaCaT cells)in a high-glucose environment and accelerated wound healing in a DFU rat model.Based on results from RNA sequencing,we defined a new microRNA(miR-4482-3p)related to the promotion of wound healing.The bioactivity of miR-4482-3p was verified by inhibiting and overexpressing miR-4482-3p.Inhibition of miR-4482-3p enhanced the migration and proliferation ability of HaCaT cells as well as the expression of vascular endothelial growth factor B(VEGFB).RLQN15 also promoted the migration and proliferation ability of HaCaT cells,and VEGFB expression was mediated via inhibition of miR-4482-3p expression by the p38 mitogen-activated protein kinase(p38MAPK)and smad3 signaling pathways.Conclusions:RL-QN15 is an effective molecule for the treatment of DFUs,with the underlying mechanism related to the inhibition of miR-4482-3p expression via the p38MAPK and smad3 signal展开更多
BACKGROUND Diabetic polyneuropathy is a very common complication of diabetes.Numerous studies are available in terms of pathogenesis.But examination methods with low reliability are still not standardized and generall...BACKGROUND Diabetic polyneuropathy is a very common complication of diabetes.Numerous studies are available in terms of pathogenesis.But examination methods with low reliability are still not standardized and generally time consuming.Highsensitive,easy-to-access methods are expected.Biochemical markers are one of the subjects of research.We aimed to discover a potential biomarker that can be used for this purpose in patients with diabetes who have not yet developed symptoms of neuropathy.AIM To determine the place and availability of visfatin and thiol-disulfide homeostasis in this disorder.METHODS A total of 392 patients with type 2 diabetes mellitus were included in the study.The polyneuropathy clinical signs were evaluated with the Subjective Peripheral Neuropathy Screen Questionnaire and Michigan Neuropathy Screening Instrument questionnaire and examination.The biochemical parameters,oxidative stress markers,visfatin,and thiol-disulfide homeostasis were analyzed and correlated with each other and clinical signs.RESULTS Subjective Peripheral Neuropathy Screen Questionnaire and Michigan Neuropathy Screening Instrument questionnaire with examination scores were correlated with each other and diabetes duration(P<0.005).Neuropathy related symptoms were present in 20.7%of the patients,but neuropathy related findings were observed in 43.9%of the patients.Serum glucose,glycated hemoglobin,and visfatin were positively correlated with each other.Also,these parameters were positively correlated with the total oxidative stress index.Total and native thiol was positively correlated with total antioxidant status and negatively with oxidant status.Inversely thiol-disulfide positively correlated with higher glucose and oxidant status and negatively with total antioxidant status(P<0.005).There was no correlation between visfatin and thiol-disulphide(P=0.092,r=0.086).However,a significant negative correlation was observed between visfatin and total with native thiol(P<0.005,r=-0.338),(P<0.005,r=-0.448).CONCLUSION Diagnosis of 展开更多
Enterovirus A71(EV-A71)is a significant human pathogen,especially in children.EV-A71 infection is one of the leading causes of hand,foot,and mouth diseases(HFMD),and can lead to neurological complications such as acut...Enterovirus A71(EV-A71)is a significant human pathogen,especially in children.EV-A71 infection is one of the leading causes of hand,foot,and mouth diseases(HFMD),and can lead to neurological complications such as acute flaccid myelitis(AFM)in severe cases.Although three EV-A71 vaccines are available in China,they are not broadly protective and have reduced efficacy against emerging strains.There is currently no approved antiviral for EV-A71.Significant progress has been made in developing antivirals against EV-A71 by targeting both viral proteins and host factors.However,viral capsid inhibitors and protease inhibitors failed in clinical trials of human rhinovirus infection due to limited efficacy or side effects.This review discusses major discoveries in EV-A71 antiviral development,analyzes the advantages and limitations of each drug target,and highlights the knowledge gaps that need to be addressed to advance the field forward.展开更多
Foot-and-mouth disease virus(FMDV) is the causative agent of a highly contagious disease in livestock. The viral proteinaseL^(rop) of FMDV is involved in pathogenicity, and mutation of theL^(rop) SAP domain reduces FM...Foot-and-mouth disease virus(FMDV) is the causative agent of a highly contagious disease in livestock. The viral proteinaseL^(rop) of FMDV is involved in pathogenicity, and mutation of theL^(rop) SAP domain reduces FMDV pathogenicity in pigs. To determine the gene expression profiles associated with decreased pathogenicity in porcine cells, we performed transcriptome analysis using next-generation sequencing technology and compared differentially expressed genes in SK6 cells infected with FMDV containingL^(rop) with either a wild-type or mutated version of the SAP domain. This analysis yielded 1,853 genes that exhibited a ≥ 2-fold change in expression and was validated by real-time quantitative PCR detection of several differentially expressed genes. Many of the differentially expressed genes correlated with antiviral responses corresponded to genes associated with transcription factors, immune regulation, cytokine production, inflammatory response, and apoptosis. Alterations in gene expression profiles may be responsible for the variations in pathogenicity observed between the two FMDV variants. Our results provided genes of interest for the further study of antiviral pathways and pathogenic mechanisms related to FMDV L^(rop).展开更多
[Objectives]To investigate the clinical effect of different doses of Xuesaitong combined with autologous platelet-rich gel(APG)on patients with diabetic foot(DF).[Methods]90 patients with diabetic foot admitted to our...[Objectives]To investigate the clinical effect of different doses of Xuesaitong combined with autologous platelet-rich gel(APG)on patients with diabetic foot(DF).[Methods]90 patients with diabetic foot admitted to our hospital from February 2017 to February 2019 were enrolled in the study.According to the random number table method,the subjects were divided into study group A and B and control group C.Group A was given a low dose of Xuesaitong combined with APG,while group B was given high-dose Xuesaitong combined with APG and group C was treated only with APG.Patients in the three groups were observed and the changes of related indexes were detected.[Results]After treatment,with regard to the three groups,the fasting blood glucose,2-h postprandial blood glucose level,HbA1c,TNF-α,Hcy,blood urea nitrogen(BUN),creatinine(Cr)and 24-h urine protein levels were all decreased,yet AT-III level was increased,and granulation tissue coverage and thickness,wound clearance rate were increased,while the repair time of ulcer surface was significantly reduced.The above indexes were all significantly different(P<0.05).Compared with the control group C,the changes of the indexes in the study group A and B were the same as above,and the difference was significant(P<0.05).Compared with the study group A,the level of AT-III in group B was significantly increased(P<0.05),while TNF-α,Hcy,BUN,Cr and 24-h urine protein levels were significantly decreased(P<0.05).There was no significant difference in the other indexes between groups A and B(P>0.05).[Conclusions]Xuesaitong combined with APG could effectively reduce the blood sugar level of DF patients,improve the clinical indexes,promote wound healing,and the high-dose group had more significant advantages and was worthy of promotion.展开更多
Mussel foot proteins(Mfps) secreted in the byssal plaque of marine mussels are widely researched for their relevance to mussel adhesion in water. As the abundant residue in the amino acid sequences of major adhesive p...Mussel foot proteins(Mfps) secreted in the byssal plaque of marine mussels are widely researched for their relevance to mussel adhesion in water. As the abundant residue in the amino acid sequences of major adhesive proteins, 3,4-dihydroxyphenylalanine(Dopa) or its catecholic moiety plays a key role in both Mfp binding to surface and cohesive cross-linking of Mfps in byssal plaques. The binding performance of an Mfp significantly depends on the content and redox state of Dopa, whereas the types of interaction vary in line with different surface chemistries and p H conditions. Thorough understanding of mussel adhesion from a molecular perspective is crucial to promote the application of synthetic mussel-bionic adhesives. This article presents a brief review of the research progress on the adhesion mechanisms of Mfps, which further emphasizes the contributions of Dopamediated interactions and considers other amino acids and factors. The involved inter-and intramolecular interactions are responsible for not only the diverse adhesion capacities of an adhesive byssal plaque as mussel's adhesion precursor but also the formation and properties of the plaque structure.展开更多
Monoclonal antibodies (McAbs) 1A9 and 9F12 against Foot-and-mouth disease virus (FMDV) serotype O were produced by fusing SP2/0 myeloma cells with splenocyte from the mouse immunized with O/China99. Both McAbs reacted...Monoclonal antibodies (McAbs) 1A9 and 9F12 against Foot-and-mouth disease virus (FMDV) serotype O were produced by fusing SP2/0 myeloma cells with splenocyte from the mouse immunized with O/China99. Both McAbs reacted with O/China99 but not with Asia 1, as determined by immunohistochemistry assay. The microneutralization titer of the McAbs 1A9 and 9F12 were 640 and 1 280, respectively. Both McAbs contain kappa light chains, but the McAbs 1A9 and 9F12 were IgG1 and IgM, respectively. In order to define the McAbs binding epitopes, the reactivity of these McAbs against VP1, P20 and P14 were examined using indirect ELISA, the result showed that both McAbs reacted with VP1 and P20. McAbs may be used for further studies of vaccine, diagnostic methods, prophylaxis, etiological and immunological researches on FMDV.展开更多
基金This work was supported by Grants from the National Key R&D Programme of China(No.2017YFD0501103 and 2017YFD0501800)the Key Development and Research Foundation of Yunnan(No.2018BB004)the Chinese Academy of Agricultural Science and Technology Innovation Project(CAAS-XTCX2016011-01 and Y2017JC55).
文摘Receptors interaction protein 2(RIP2)is a specific adaptor molecule in the downstream of NOD2.The role of RIP2 during foot-and-mouth disease virus(FMDV)infection remains unknown.Here,our results showed that RIP2 inhibited FMDV replication and played an important role in the activation of IFN-βand NF-κB signal pathways during FMDV infection.FMDV infection triggered RIP2 transcription,while it reduced the expression of RIP2 protein.Detailed analysis showed that FMDV 2B,2C,3C^(pro),and L^(pro) proteins were responsible for inducing the reduction of RIP2 protein.3C^(pro) and L^(pro) are viral proteinases that can induce the cleavage or reduction of many host proteins and block host protein synthesis.The carboxyl terminal 105-C114 and 135-C144 regions of 2B were essential for reduction of RIP2.Our results also showed that the N terminal 1-61 region of 2C were essential for the reduction of RIP2.The 2C-induced reduction of RIP2 was dependent on inducing the reduction of poly(A)-binding protein 1(PABPC1).The interaction between RIP2 and 2C was observed in the context of viral infection,and the residues 1-61 were required for the interaction.These data clarify novel mechanisms of reduction of RIP2 mediated by FMDV.
基金supported by grants from the National Natural Science Foundation of China(32360138,32060212,32301054 and 81760648)Key Program of Yunnan Fundamental Research Project(202301AS070036)+1 种基金Outstanding Youth Program of Yunnan Applied Basic Research Project-Kunming Medical University Union Foundation(202301AY070001-301)Project of Yunnan Applied Basic Research Project-Kunming Medical University Union Foundation(202101AY070001-006 and 202101AY070001-036).
文摘Background:Wound management of diabetic foot ulcers(DFUs)is a complex and challenging task,and existing strategies fail to meet clinical needs.Therefore,it is important to develop novel drug candidates and discover new therapeutic targets.However,reports on peptides as molecular probes for resolving issues related to DFUs remain rare.This study utilized peptide RL-QN15 as an exogenous molecular probe to investigate the underlying mechanism of endogenous non-coding RNA in DFU wound healing.The aim was to generate novel insights for the clinical management of DFUs and identify potential drug targets.Methods:We investigated the wound-healing efficiency of peptide RL-QN15 under diabetic con-ditions using in vitro and in vivo experimental models.RNA sequencing,in vitro transfection,quantitative real-time polymerase chain reaction,western blotting,dual luciferase reporter gene detection,in vitro cell scratches,and cell proliferation and migration assays were performed to explore the potential mechanism underlying the promoting effects of RL-QN15 on DFU repair.Results:Peptide RL-QN15 enhanced the migration and proliferation of human immortalized keratinocytes(HaCaT cells)in a high-glucose environment and accelerated wound healing in a DFU rat model.Based on results from RNA sequencing,we defined a new microRNA(miR-4482-3p)related to the promotion of wound healing.The bioactivity of miR-4482-3p was verified by inhibiting and overexpressing miR-4482-3p.Inhibition of miR-4482-3p enhanced the migration and proliferation ability of HaCaT cells as well as the expression of vascular endothelial growth factor B(VEGFB).RLQN15 also promoted the migration and proliferation ability of HaCaT cells,and VEGFB expression was mediated via inhibition of miR-4482-3p expression by the p38 mitogen-activated protein kinase(p38MAPK)and smad3 signaling pathways.Conclusions:RL-QN15 is an effective molecule for the treatment of DFUs,with the underlying mechanism related to the inhibition of miR-4482-3p expression via the p38MAPK and smad3 signal
基金Research and Agricultural Project of Zhejiang Technology Department(2008C22026,2009C32016)Key Project of Zhejiang Education Department(20070430)Xin-Miao Project of Zhejiang Technology Department(2008R40G2110002)
文摘BACKGROUND Diabetic polyneuropathy is a very common complication of diabetes.Numerous studies are available in terms of pathogenesis.But examination methods with low reliability are still not standardized and generally time consuming.Highsensitive,easy-to-access methods are expected.Biochemical markers are one of the subjects of research.We aimed to discover a potential biomarker that can be used for this purpose in patients with diabetes who have not yet developed symptoms of neuropathy.AIM To determine the place and availability of visfatin and thiol-disulfide homeostasis in this disorder.METHODS A total of 392 patients with type 2 diabetes mellitus were included in the study.The polyneuropathy clinical signs were evaluated with the Subjective Peripheral Neuropathy Screen Questionnaire and Michigan Neuropathy Screening Instrument questionnaire and examination.The biochemical parameters,oxidative stress markers,visfatin,and thiol-disulfide homeostasis were analyzed and correlated with each other and clinical signs.RESULTS Subjective Peripheral Neuropathy Screen Questionnaire and Michigan Neuropathy Screening Instrument questionnaire with examination scores were correlated with each other and diabetes duration(P<0.005).Neuropathy related symptoms were present in 20.7%of the patients,but neuropathy related findings were observed in 43.9%of the patients.Serum glucose,glycated hemoglobin,and visfatin were positively correlated with each other.Also,these parameters were positively correlated with the total oxidative stress index.Total and native thiol was positively correlated with total antioxidant status and negatively with oxidant status.Inversely thiol-disulfide positively correlated with higher glucose and oxidant status and negatively with total antioxidant status(P<0.005).There was no correlation between visfatin and thiol-disulphide(P=0.092,r=0.086).However,a significant negative correlation was observed between visfatin and total with native thiol(P<0.005,r=-0.338),(P<0.005,r=-0.448).CONCLUSION Diagnosis of
基金This research was supported by the National Institute of Allergy and Infectious Diseasess of Health(NIH,USA,grants AI147325 and AI157046)the Arizona Biomedical Research Commission Centre Young Investigator grant(ADHS18-198859,USA)to Jun WangYanmei Hu was supported by the NIH training grant T32 GM008804(USA).
文摘Enterovirus A71(EV-A71)is a significant human pathogen,especially in children.EV-A71 infection is one of the leading causes of hand,foot,and mouth diseases(HFMD),and can lead to neurological complications such as acute flaccid myelitis(AFM)in severe cases.Although three EV-A71 vaccines are available in China,they are not broadly protective and have reduced efficacy against emerging strains.There is currently no approved antiviral for EV-A71.Significant progress has been made in developing antivirals against EV-A71 by targeting both viral proteins and host factors.However,viral capsid inhibitors and protease inhibitors failed in clinical trials of human rhinovirus infection due to limited efficacy or side effects.This review discusses major discoveries in EV-A71 antiviral development,analyzes the advantages and limitations of each drug target,and highlights the knowledge gaps that need to be addressed to advance the field forward.
基金supported by grants from the National Science and Technology Ministry (2015BAD12B04)National Natural Sciences Foundation of China (No.31302118,31502042 and 31402179)+2 种基金the Gansu Science Foundation for Distinguished Young Scholars (no.145RJDA328)the International Atomic Energy Agency (16025/R0)the Key technologies R&Dprogram of Gansu Province (1302NKDA027)
文摘Foot-and-mouth disease virus(FMDV) is the causative agent of a highly contagious disease in livestock. The viral proteinaseL^(rop) of FMDV is involved in pathogenicity, and mutation of theL^(rop) SAP domain reduces FMDV pathogenicity in pigs. To determine the gene expression profiles associated with decreased pathogenicity in porcine cells, we performed transcriptome analysis using next-generation sequencing technology and compared differentially expressed genes in SK6 cells infected with FMDV containingL^(rop) with either a wild-type or mutated version of the SAP domain. This analysis yielded 1,853 genes that exhibited a ≥ 2-fold change in expression and was validated by real-time quantitative PCR detection of several differentially expressed genes. Many of the differentially expressed genes correlated with antiviral responses corresponded to genes associated with transcription factors, immune regulation, cytokine production, inflammatory response, and apoptosis. Alterations in gene expression profiles may be responsible for the variations in pathogenicity observed between the two FMDV variants. Our results provided genes of interest for the further study of antiviral pathways and pathogenic mechanisms related to FMDV L^(rop).
基金Applied Basic Research Program of Yunnan Province(2017FH001-081).
文摘[Objectives]To investigate the clinical effect of different doses of Xuesaitong combined with autologous platelet-rich gel(APG)on patients with diabetic foot(DF).[Methods]90 patients with diabetic foot admitted to our hospital from February 2017 to February 2019 were enrolled in the study.According to the random number table method,the subjects were divided into study group A and B and control group C.Group A was given a low dose of Xuesaitong combined with APG,while group B was given high-dose Xuesaitong combined with APG and group C was treated only with APG.Patients in the three groups were observed and the changes of related indexes were detected.[Results]After treatment,with regard to the three groups,the fasting blood glucose,2-h postprandial blood glucose level,HbA1c,TNF-α,Hcy,blood urea nitrogen(BUN),creatinine(Cr)and 24-h urine protein levels were all decreased,yet AT-III level was increased,and granulation tissue coverage and thickness,wound clearance rate were increased,while the repair time of ulcer surface was significantly reduced.The above indexes were all significantly different(P<0.05).Compared with the control group C,the changes of the indexes in the study group A and B were the same as above,and the difference was significant(P<0.05).Compared with the study group A,the level of AT-III in group B was significantly increased(P<0.05),while TNF-α,Hcy,BUN,Cr and 24-h urine protein levels were significantly decreased(P<0.05).There was no significant difference in the other indexes between groups A and B(P>0.05).[Conclusions]Xuesaitong combined with APG could effectively reduce the blood sugar level of DF patients,improve the clinical indexes,promote wound healing,and the high-dose group had more significant advantages and was worthy of promotion.
基金supported by the National Natural Science Foundation of China (Grant No. 51605090)the Natural Science Foundation of Jiangsu Province (Grant Nos. BK20160670 and BK20160776)the Fundamental Research Funds for the Central Universities (Grant No.2242019k1G011)。
文摘Mussel foot proteins(Mfps) secreted in the byssal plaque of marine mussels are widely researched for their relevance to mussel adhesion in water. As the abundant residue in the amino acid sequences of major adhesive proteins, 3,4-dihydroxyphenylalanine(Dopa) or its catecholic moiety plays a key role in both Mfp binding to surface and cohesive cross-linking of Mfps in byssal plaques. The binding performance of an Mfp significantly depends on the content and redox state of Dopa, whereas the types of interaction vary in line with different surface chemistries and p H conditions. Thorough understanding of mussel adhesion from a molecular perspective is crucial to promote the application of synthetic mussel-bionic adhesives. This article presents a brief review of the research progress on the adhesion mechanisms of Mfps, which further emphasizes the contributions of Dopamediated interactions and considers other amino acids and factors. The involved inter-and intramolecular interactions are responsible for not only the diverse adhesion capacities of an adhesive byssal plaque as mussel's adhesion precursor but also the formation and properties of the plaque structure.
基金The national high technology research and development program of China 863 (2006AA10A204)The national science and technology pillar program (2006BAD06A17)
文摘Monoclonal antibodies (McAbs) 1A9 and 9F12 against Foot-and-mouth disease virus (FMDV) serotype O were produced by fusing SP2/0 myeloma cells with splenocyte from the mouse immunized with O/China99. Both McAbs reacted with O/China99 but not with Asia 1, as determined by immunohistochemistry assay. The microneutralization titer of the McAbs 1A9 and 9F12 were 640 and 1 280, respectively. Both McAbs contain kappa light chains, but the McAbs 1A9 and 9F12 were IgG1 and IgM, respectively. In order to define the McAbs binding epitopes, the reactivity of these McAbs against VP1, P20 and P14 were examined using indirect ELISA, the result showed that both McAbs reacted with VP1 and P20. McAbs may be used for further studies of vaccine, diagnostic methods, prophylaxis, etiological and immunological researches on FMDV.
