Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located...Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located in ZIKV 5'UTR and virus production.Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining,Subsequently,liquid chromatographytandem mass spectrometry(LC-MS/MS),bioinformatics analysis,and Western blot were used to identify the candidate proteins binding to ZIKV 5'UTR.Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production,respecitvely.Results tRSA RNA pull-down assay,LC-MS/MS,and Western blot analysis showed that polypyrimidine tractbinding protein(PTB)bound to the ZIKV 5'UTR.Furthermore,dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV(t=10.220,P<0.001),while PTB knockdown had the opposite effect(t=4.897,P<0.01).Additionally,virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer(t=6.400,P<0.01),whereas reducing PTB expression level weakened virus infectivity(t=5.055,P<0.01).Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.展开更多
The past twenty years have seen the increasingly important role of ontology in traditional Chinese medicine(TCM).However,the development of TCM ontology faces many challenges.Since the epistemologies dramatically diff...The past twenty years have seen the increasingly important role of ontology in traditional Chinese medicine(TCM).However,the development of TCM ontology faces many challenges.Since the epistemologies dramatically differ between TCM and contemporary biomedicine,it is hard to apply the existing top-level ontology mechanically.“Data silos”are widely present in the currently available terminology standards,term sets,and ontologies.The formal representation of ontology needs to be further improved in TCM.Therefore,we propose a unified basic semantic framework of TCM based on in-depth theoretical research on the existing top・level ontology and a re-study of important concepts in TCM.Under such a framework,ontologies in TCM subdomains should be built collaboratively and be represented formally in a common format.Besides,extensive cooperation should be encouraged by establishing ontology research communities to promote ontology peer review and reuse.展开更多
The response to endoplasmic reticulum(ER)stress has been noted in both human depression cases and depression models in rodents.Wuling powder,derived from the mycelium of the esteemed fungus Xylaria Nigripes(Kl.)Sacc,h...The response to endoplasmic reticulum(ER)stress has been noted in both human depression cases and depression models in rodents.Wuling powder,derived from the mycelium of the esteemed fungus Xylaria Nigripes(Kl.)Sacc,has demonstrated efficacy in alleviating depressive symptoms.The purpose of this research was to explore the antidepressant properties of Wuling powder and its basic molecular effects,particularly regarding alterations in ER stress.A model of social defeat stress was created by introducing a mouse to the cage of an unfamiliar,hostile mouse for intervals of 5–10 min daily over a span of 10 d.Subsequently,the mice received oral doses of Wuling powder for 2 weeks.The social approach-avoidance assay was employed to evaluate signs of depression-like behaviors.Moreover,protein and gene expressions linked to ER stress triggered by social defeat were analyzed through Western blotting analysis and quantitative real-time PCR.The behavioral tests indicated that Wuling powder ameliorated behaviors associated with depression due to social defeat stress.Treatment with Wuling powder markedly reduced the increased levels of the 78-k Da glucose-regulated protein and protein disulfide isomerase caused by social defeat stress.It also diminished the expression of inositol-requiring enzyme 1α(IRE1α)and spliced X box-binding protein-1(s XBP1)at the protein and m RNA levels.Furthermore,Wuling treatment notably decreased the levels of phosphorylated eukaryotic initiation factor 2 alpha kinase(P-e IF2α),activating transcription factor 4(ATF4)and C/EBP homologous protein(CHOP),simultaneously enhancing the ratio of B-cell lymphoma 2(Bcl-2)to Bcl-2-associated X protein(Bax).These results suggested that Wuling powder could alleviate ER stress and inhibit cell apoptosis in the hippocampus by inhibiting protein translation and synthesis,thereby attenuating depressive-like behavior.展开更多
In the present study,we aimed to evaluate the anti-inflammatory mechanism of galanthamine,a classic therapeutic drug for Alzheimer s disease(AD).The co-culture system of hippocampal nerve cell line HT-22 and microglia...In the present study,we aimed to evaluate the anti-inflammatory mechanism of galanthamine,a classic therapeutic drug for Alzheimer s disease(AD).The co-culture system of hippocampal nerve cell line HT-22 and microglial cell line BV-2 was established to observe the effect of galanthamine on the expressions of inflammatory factors induced by lipopolysaccharide(LP S).MTT method was used to observe the protective effect of galanthamine on neurons.ELISA and qPCR methods were used to detect the expressions of interleukin-β(IL-1β) and IL-1 receptor antagonist(IL-1 RA) at the protein and mRNA levels,respectively.IL-1β and IL-1 RA were evaluated by the ELISA method after pretreating with galanthamine and α7 nAChR blockerα-bungarotoxin(α-bun),mAChR blocker atropine(Atr),PI3 K inhibitor LY294002,IKKβ inhibitor SC514,or MEK inhibitor PD98059,respectively.The results showed that galanthamine significantly inhibited LPS-induced increased IL-1β and IL-1 RA expressions and maintained the ratio of IL-1β/IL-1 RA.α-Bun could block the regulatory effect of galanthamine on IL-1β and IL-1 RA.As PI3 K and NF-κB pathways were blocked,the regulatory effect of galanthamine on the IL-1β expression was significantly inhibited.Blocking PI3 K and MEK pathways could significantly inhibit the regulation of galanthamine on IL-1 RA expression.In summary,galanthamine regulated the inflammatory activity of the IL-1 subfamily to play an anti-inflammatory role mediated by α7 nAChR and PI3 K/NF-κB/MAPK pathways,which probably provided a new strategy for AD treatment.展开更多
文摘Objectives To identify the 5'untranslated region of Zika virus(ZIKV 5'UTR)RNA-binding proteins and to investigate the impact of the binding protein on the activity of internal ribosomal entry site(IRES)located in ZIKV 5'UTR and virus production.Methods Interacting proteins in U251 cells were captured using tRSA-tagged ZIKV 5'UTR RNA and tRSA-ZIKV 5'UTR RNA-binding proteins were visualized by SDS-PAGE silver staining,Subsequently,liquid chromatographytandem mass spectrometry(LC-MS/MS),bioinformatics analysis,and Western blot were used to identify the candidate proteins binding to ZIKV 5'UTR.Dicistronic expression assay and plaque forming assay were performed to analyze the effect of the binding protein on ZIKV IRES activity and ZIKV production,respecitvely.Results tRSA RNA pull-down assay,LC-MS/MS,and Western blot analysis showed that polypyrimidine tractbinding protein(PTB)bound to the ZIKV 5'UTR.Furthermore,dual luciferase reporter assay revealed that overexpression of PTB significantly enhanced the IRES activity of ZIKV(t=10.220,P<0.001),while PTB knockdown had the opposite effect(t=4.897,P<0.01).Additionally,virus plaque forming assay demonstrated that up-regulation of PTB expression significantly enhanced viral titer(t=6.400,P<0.01),whereas reducing PTB expression level weakened virus infectivity(t=5.055,P<0.01).Conclusion PTB positively interacts with the ZIKV 5'UTR and enhances IRES activity and virus production.
文摘The past twenty years have seen the increasingly important role of ontology in traditional Chinese medicine(TCM).However,the development of TCM ontology faces many challenges.Since the epistemologies dramatically differ between TCM and contemporary biomedicine,it is hard to apply the existing top-level ontology mechanically.“Data silos”are widely present in the currently available terminology standards,term sets,and ontologies.The formal representation of ontology needs to be further improved in TCM.Therefore,we propose a unified basic semantic framework of TCM based on in-depth theoretical research on the existing top・level ontology and a re-study of important concepts in TCM.Under such a framework,ontologies in TCM subdomains should be built collaboratively and be represented formally in a common format.Besides,extensive cooperation should be encouraged by establishing ontology research communities to promote ontology peer review and reuse.
基金National Natural Science Foundation of China(Grant No.81100801 and 81050025)the foundation of Zhejiang Jolly Pharmaceutical Co.,Ltd.
文摘The response to endoplasmic reticulum(ER)stress has been noted in both human depression cases and depression models in rodents.Wuling powder,derived from the mycelium of the esteemed fungus Xylaria Nigripes(Kl.)Sacc,has demonstrated efficacy in alleviating depressive symptoms.The purpose of this research was to explore the antidepressant properties of Wuling powder and its basic molecular effects,particularly regarding alterations in ER stress.A model of social defeat stress was created by introducing a mouse to the cage of an unfamiliar,hostile mouse for intervals of 5–10 min daily over a span of 10 d.Subsequently,the mice received oral doses of Wuling powder for 2 weeks.The social approach-avoidance assay was employed to evaluate signs of depression-like behaviors.Moreover,protein and gene expressions linked to ER stress triggered by social defeat were analyzed through Western blotting analysis and quantitative real-time PCR.The behavioral tests indicated that Wuling powder ameliorated behaviors associated with depression due to social defeat stress.Treatment with Wuling powder markedly reduced the increased levels of the 78-k Da glucose-regulated protein and protein disulfide isomerase caused by social defeat stress.It also diminished the expression of inositol-requiring enzyme 1α(IRE1α)and spliced X box-binding protein-1(s XBP1)at the protein and m RNA levels.Furthermore,Wuling treatment notably decreased the levels of phosphorylated eukaryotic initiation factor 2 alpha kinase(P-e IF2α),activating transcription factor 4(ATF4)and C/EBP homologous protein(CHOP),simultaneously enhancing the ratio of B-cell lymphoma 2(Bcl-2)to Bcl-2-associated X protein(Bax).These results suggested that Wuling powder could alleviate ER stress and inhibit cell apoptosis in the hippocampus by inhibiting protein translation and synthesis,thereby attenuating depressive-like behavior.
基金National Natural Science Foundation of China (Grant No. 81100801)Peking Union Medical College Small-Scale Characteristic School Education Project。
文摘In the present study,we aimed to evaluate the anti-inflammatory mechanism of galanthamine,a classic therapeutic drug for Alzheimer s disease(AD).The co-culture system of hippocampal nerve cell line HT-22 and microglial cell line BV-2 was established to observe the effect of galanthamine on the expressions of inflammatory factors induced by lipopolysaccharide(LP S).MTT method was used to observe the protective effect of galanthamine on neurons.ELISA and qPCR methods were used to detect the expressions of interleukin-β(IL-1β) and IL-1 receptor antagonist(IL-1 RA) at the protein and mRNA levels,respectively.IL-1β and IL-1 RA were evaluated by the ELISA method after pretreating with galanthamine and α7 nAChR blockerα-bungarotoxin(α-bun),mAChR blocker atropine(Atr),PI3 K inhibitor LY294002,IKKβ inhibitor SC514,or MEK inhibitor PD98059,respectively.The results showed that galanthamine significantly inhibited LPS-induced increased IL-1β and IL-1 RA expressions and maintained the ratio of IL-1β/IL-1 RA.α-Bun could block the regulatory effect of galanthamine on IL-1β and IL-1 RA.As PI3 K and NF-κB pathways were blocked,the regulatory effect of galanthamine on the IL-1β expression was significantly inhibited.Blocking PI3 K and MEK pathways could significantly inhibit the regulation of galanthamine on IL-1 RA expression.In summary,galanthamine regulated the inflammatory activity of the IL-1 subfamily to play an anti-inflammatory role mediated by α7 nAChR and PI3 K/NF-κB/MAPK pathways,which probably provided a new strategy for AD treatment.
