Objective:To evaluate the immunological response elicited by an inactivated bacterial vector carrying the K39 antigen of Leishmania infantum,and a purified antigen.Methods:Mice were subjected to the following treatmen...Objective:To evaluate the immunological response elicited by an inactivated bacterial vector carrying the K39 antigen of Leishmania infantum,and a purified antigen.Methods:Mice were subjected to the following treatments:(1)Purified recombinant K39(rK39)protein at a 20μg dose with complete Freund’s adjuvant;(2)Inactivated Escherichia coli(BL21 DE3)carrying the K39 protein at an equivalent total protein content of 200μg;(3)Inactivated bacteria lacking the K39 protein;(4)Non-immunized control animals.Serological monitoring was performed.All groups were challenged by intraperitoneal injection of 10^(7) Leishmania infantum promastigotes.After euthanasia,the liver and spleen were collected to analyze the levels of TNF,IFN-γ,IL-12,IL-4,and IL-10.Results:Mice immunized with purified rK39 or the inactivated bacterial vector carrying the K39 antigen of Leishmania infantum showed a long-lasting immune response with high levels of polyclonal antibodies specifically recognizing the recombinant proteins.The IgG1 subclass was the predominant immunoglobulin;however,the induction of IgG2a and the profile of cytokines produced were indicative of the induction of a mixed-type response.Conclusions:The inactivated bacterial vector carrying the K39 antigen,as well as the purified antigen can induce a long-lasting immune response in immunized mice,predominantly favouring a Th2 profile response.展开更多
Activated G-protein-coupled receptor 39(GPR39)has been shown to attenuate inflammation by interacting with sirtuin 1(SIRT1)and peroxisome proliferator-activated receptor-γcoactivator 1α(PGC-1α).However,whether GPR3...Activated G-protein-coupled receptor 39(GPR39)has been shown to attenuate inflammation by interacting with sirtuin 1(SIRT1)and peroxisome proliferator-activated receptor-γcoactivator 1α(PGC-1α).However,whether GPR39 attenuates neuropathic pain remains unclear.In this study,we established a Sprague-Dawley rat model of spared nerve injury-induced neuropathic pain and found that GPR39 expression was significantly decreased in neurons and microglia in the spinal dorsal horn compared with sham-operated rats.Intrathecal injection of TC-G 1008,a specific agonist of GPR39,significantly alleviated mechanical allodynia in the rats with spared nerve injury,improved spinal cord mitochondrial biogenesis,and alleviated neuroinflammation.These changes were abolished by GPR39 small interfering RNA(siRNA),Ex-527(SIRT1 inhibitor),and PGC-1αsiRNA.Taken together,these findings show that GPR39 activation ameliorates mechanical allodynia by activating the SIRT1/PGC-1αpathway in rats with spared nerve injury.展开更多
The Jitang metamorphic complex is key to studying the tectonic evolution of the Northern Lancangjiang zone.Through structural-lithological mapping,structural analysis and laboratory testing,the composition of the Jita...The Jitang metamorphic complex is key to studying the tectonic evolution of the Northern Lancangjiang zone.Through structural-lithological mapping,structural analysis and laboratory testing,the composition of the Jitang metamorphic complex was determined.The macro-and microstructural analyses of the ductile detachment shear zone(Guoxuepu ductile shear zone,2–4 km wide)between the metamorphic complex and the overlying sedimentary cap show that the shear sense of the ductile shear zones is top-to-the-southeast.The presence of various deformation features and quartz C-axis electron backscatter diffraction(EBSD)fabric analysis suggests multiple deformation events occurring at different temperatures.The average stress is 25.68 MPa,with the strain rates(έ)ranging from 9.77×10^(−14)s^(−1)to 6.52×10^(−16)s^(−1).The finite strain of the Guoxuepu ductile shear zone indicates an elongated strain pattern.The average kinematic vorticity of the Guoxuepu ductile shear zone is 0.88,implying that the shear zone is dominated by simple shear.The muscovite selected from the protomylonite samples in the Guoxuepu ductile shear zone yields a 40Ar-39Ar age of 60.09±0.38 Ma.It is suggested that,coeval with the initial Indo–Eurasian collision,the development of strike-slip faults led to a weak and unstable crust,upwelling of lower crust magma,then induced the detachment of the Jitang metamorphic complex in the Eocene.展开更多
文摘目的探究DDX解旋酶39A(DEAD-box RNA helicases 39A,DDX39A)对食管鳞癌KYSE-150和TE-1细胞生物学行为的影响及作用机制。方法利用癌症基因组图谱(the cancer genome atlas,TCGA)和高通量基因表达数据库(gene expression omnibus,GEO)分析DDX39A在食管鳞癌肿瘤组织和正常组织中的表达差异。进一步在TCGA数据库中,采用Spearman相关分析(设置r>0.5,P<0.05)得到与DDX39A表达最相关基因,通过京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)和基因集富集分析(gene set enrichment analysis,GSEA),注释DDX39A在食管鳞癌中的生物学功能。通过慢病毒介导的短发夹RNA(shRNA)干扰技术下调DDX39A在食管鳞癌KYSE-150和TE-1细胞中的表达,并将细胞分为对照(shCtrl)组和DDX39A敲减(shDDX39A)组。利用qPCR和Western blot法检测敲减效率;利用Celigo法、克隆形成实验和MTT法检测KYSE-150和TE-1细胞的增殖能力和活力;利用流式细胞术检测KYSE-150和TE-1细胞的凋亡水平;利用Transwell实验检测KYSE-150和TE-1细胞的迁移和侵袭能力;利用裸鼠成瘤实验检测敲减DDX39A对体内肿瘤的作用。利用Western blot法筛选敲减DDX39A后KYSE-150细胞内蛋白表达水平显著变化的肿瘤相关经典通路分子。通过慢病毒转染法构建已筛选的分子CDH2(Cadherin 2)过表达的稳转细胞系,并将KYSE-150细胞分为shDDX39A+NC-OE组(转染shDDX39A慢病毒和过表达空载对照病毒)和shDDX39A+CDH2-OE组(转染shDDX39A慢病毒和过表达CDH2病毒)。利用Celigo法、MTT法和Transwell实验分别检测过表达CDH2对DDX39A敲减细胞增殖能力、活力以及转移能力的影响。结果与正常组织相比,食管鳞癌组织中DDX39A的表达显著上调(P<0.001)。DDX39A及其相关基因主要作用于遗传信息的翻译与加工和细胞凋亡等过程。与shCtrl组相比,shDDX39A组KYSE-150和TE-1细胞中DDX39A的mRNA和蛋白表达显著降低(P<0.01),KYSE-150和TE-1细胞增殖和细胞活力显著下降(P<0.001),KYSE-150和TE-1细胞的克隆形成能力显著降低(P<0.01),KYSE-150和TE-1细胞的凋亡水平显著升高(P<0.01),KYSE-150和TE-1细胞的迁移和侵袭能力显著下降(P<0.001)。裸鼠成瘤实验提示DDX39A敲减后瘤体的体积和质量均明显减少(P<0.001)。与shDDX39A+NC-OE组相比,shDDX39A+CDH2-OE组KYSE-150细胞的增殖、细胞活力以及转移能力均显著提高(P<0.001)。结论DDX39A可作为食管鳞癌诊治的新靶点,上调其表达可能通过诱导上皮间充质转化促进食管鳞癌细胞生长、迁移与侵袭。
基金supported by grants from the Brazilian Agencies:Coordenação de Aperfeiçoamento de Pessoal de Nível Superior(CAPES-Financial code 001)Conselho Nacional de Desenvolvimento Científico e Tecnológico(CNPq)Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico(FUNCAP).
文摘Objective:To evaluate the immunological response elicited by an inactivated bacterial vector carrying the K39 antigen of Leishmania infantum,and a purified antigen.Methods:Mice were subjected to the following treatments:(1)Purified recombinant K39(rK39)protein at a 20μg dose with complete Freund’s adjuvant;(2)Inactivated Escherichia coli(BL21 DE3)carrying the K39 protein at an equivalent total protein content of 200μg;(3)Inactivated bacteria lacking the K39 protein;(4)Non-immunized control animals.Serological monitoring was performed.All groups were challenged by intraperitoneal injection of 10^(7) Leishmania infantum promastigotes.After euthanasia,the liver and spleen were collected to analyze the levels of TNF,IFN-γ,IL-12,IL-4,and IL-10.Results:Mice immunized with purified rK39 or the inactivated bacterial vector carrying the K39 antigen of Leishmania infantum showed a long-lasting immune response with high levels of polyclonal antibodies specifically recognizing the recombinant proteins.The IgG1 subclass was the predominant immunoglobulin;however,the induction of IgG2a and the profile of cytokines produced were indicative of the induction of a mixed-type response.Conclusions:The inactivated bacterial vector carrying the K39 antigen,as well as the purified antigen can induce a long-lasting immune response in immunized mice,predominantly favouring a Th2 profile response.
基金supported by the National Notural Science Foundation of China,Nos.82071556 and 82271291 (both to WM)
文摘Activated G-protein-coupled receptor 39(GPR39)has been shown to attenuate inflammation by interacting with sirtuin 1(SIRT1)and peroxisome proliferator-activated receptor-γcoactivator 1α(PGC-1α).However,whether GPR39 attenuates neuropathic pain remains unclear.In this study,we established a Sprague-Dawley rat model of spared nerve injury-induced neuropathic pain and found that GPR39 expression was significantly decreased in neurons and microglia in the spinal dorsal horn compared with sham-operated rats.Intrathecal injection of TC-G 1008,a specific agonist of GPR39,significantly alleviated mechanical allodynia in the rats with spared nerve injury,improved spinal cord mitochondrial biogenesis,and alleviated neuroinflammation.These changes were abolished by GPR39 small interfering RNA(siRNA),Ex-527(SIRT1 inhibitor),and PGC-1αsiRNA.Taken together,these findings show that GPR39 activation ameliorates mechanical allodynia by activating the SIRT1/PGC-1αpathway in rats with spared nerve injury.
基金financially supported by the China Geological Survey Scientific Research Project(Grant Nos.DD20190167 and DD20190053)the National Natural Science Foundation of China(Grant No.42172259).
文摘The Jitang metamorphic complex is key to studying the tectonic evolution of the Northern Lancangjiang zone.Through structural-lithological mapping,structural analysis and laboratory testing,the composition of the Jitang metamorphic complex was determined.The macro-and microstructural analyses of the ductile detachment shear zone(Guoxuepu ductile shear zone,2–4 km wide)between the metamorphic complex and the overlying sedimentary cap show that the shear sense of the ductile shear zones is top-to-the-southeast.The presence of various deformation features and quartz C-axis electron backscatter diffraction(EBSD)fabric analysis suggests multiple deformation events occurring at different temperatures.The average stress is 25.68 MPa,with the strain rates(έ)ranging from 9.77×10^(−14)s^(−1)to 6.52×10^(−16)s^(−1).The finite strain of the Guoxuepu ductile shear zone indicates an elongated strain pattern.The average kinematic vorticity of the Guoxuepu ductile shear zone is 0.88,implying that the shear zone is dominated by simple shear.The muscovite selected from the protomylonite samples in the Guoxuepu ductile shear zone yields a 40Ar-39Ar age of 60.09±0.38 Ma.It is suggested that,coeval with the initial Indo–Eurasian collision,the development of strike-slip faults led to a weak and unstable crust,upwelling of lower crust magma,then induced the detachment of the Jitang metamorphic complex in the Eocene.
