Morphological and functional abnormalities of vascular endothelial cells(VECs) are risk factors of ischemiareperfusion in skin flaps.Signaling pathway mediated by interleukin-1 receptor(IL-1 R) is essential to hypoxia...Morphological and functional abnormalities of vascular endothelial cells(VECs) are risk factors of ischemiareperfusion in skin flaps.Signaling pathway mediated by interleukin-1 receptor(IL-1 R) is essential to hypoxia/reoxygenation(H/R) injury of VECs.While the TIR/BB-loop mimetic(AS-1) disrupts the interaction between IL-1 R and myeloid differentiation primary-response protein 88(MyD88),its role in the VECs dysfunction under H/R is unclear.In this study,we first showed that there was an infiltration of inflammatory cells and the apoptosis of VECs by using a skin flap section from patients who received flap transplantation.We then showed that the H/R treatment induced apoptosis and loss of cell migration of endothelial cell line H926 were attenuated by AS-1.Furthermore,our data suggested that AS-1 inhibits the interaction between IL-1 R and MyD88,and subsequent phosphorylation of IκB and p38 pathway,as well as the nuclear localization of NF-κB subunit p65/p50.Thus,this study indicated that the protective role of AS-1 in H/R induced cellular injury may be due to the AS-1 mediated down-regulation of IL-1 R signaling pathway.展开更多
植物果聚糖是一类重要的可溶性碳水化合物,其在植物中的积累可提高植物的抗逆性。为了解大蒜蔗糖:蔗糖1-果糖基转移酶的序列特征和功能,本研究采用TA克隆方法(Original TA Cloning Kit)得到乐都紫皮大蒜As-1-SST基因全长序列,利用BLAST...植物果聚糖是一类重要的可溶性碳水化合物,其在植物中的积累可提高植物的抗逆性。为了解大蒜蔗糖:蔗糖1-果糖基转移酶的序列特征和功能,本研究采用TA克隆方法(Original TA Cloning Kit)得到乐都紫皮大蒜As-1-SST基因全长序列,利用BLAST、DNAMAN、ProtParam、SWISS-MODEL、MEGA等生物信息工具分析其序列特征,通过荧光定量PCR(qRT-PCR)分析As-1-SST基因在大蒜根、假茎、叶片和鳞芽中的表达差异及其对低温和干旱胁迫的响应情况。结果表明,大蒜As-1-SST基因全长1872 bp,编码623个氨基酸,推测蛋白质分子质量为69.76 kDa,理论等电点为5.19,为不稳定亲水性蛋白;亚细胞定位预测结果显示,As-1-SST蛋白主要定位于液泡,该蛋白无信号肽,包含2个特异位点,属于糖苷水解酶32(GH32)家族。在进化关系上,大蒜As-1-SST与百合科的洋葱1-SST亲缘关系最为接近。qRT-PCR分析表明,As-1-SST基因在根中的表达量最高,其次是假茎,在鳞芽和叶片中表达水平较低,具有明显的组织特异性;不同组织As-1-SST对于低温及干旱胁迫的响应差异显著,低温胁迫显著诱导了根、假茎、叶片中As-1-SST的表达,而干旱胁迫只显著提高了鳞芽中As-1-SST的表达量,说明大蒜各组织As-1-SST对逆境信号的响应机制不同。本研究为进一步鉴定大蒜果聚糖合成酶基因的生物信息学功能和表达调控机制提供了一定的理论依据。展开更多
Objective： To test whether IL-1 RI/My088-TIR mimic AS-1 can work as a new compound that targeted at blocking MyD88- dependent signaling pathway, we investigated the physical structure and biological function of AS-1....Objective： To test whether IL-1 RI/My088-TIR mimic AS-1 can work as a new compound that targeted at blocking MyD88- dependent signaling pathway, we investigated the physical structure and biological function of AS-1. Methods：The crystallographic structure of AS-1 was examined by 1^H nuclear magnetic resonance. The toxicity of AS-1 was measured with Methyl thiazolyl tetrazolium （MTT） assay. The effect of AS-1 on phosphorylation state of p38 MAPK and IRAK-1 was observed with Western blot. Results：The crystallographic details of AS-1 demonstrated that it was a tri-peptide sequence[（F/Y）-（V/L/I）-（P/G）] of the IL-1R I -TIR domain BBloop. No toxicity of AS-1 was shown to HEK 293A cells. The phosphorylation of p38 MAPK, induced by IL-1β significantly increased from those in the control group. AS-1 significantly reduced the phosphorylation of p38 MAPK induced by IL-1β. IL-1β increased the phosphorylation of IRAK-1 significantly, which was prevented by AS-1. Conclusion：AS-1 is a competitive mimic between IL-1R I-TIR and MyD88-TIR domain, which most likely interferes with MyD88-dependent signaling pathway.展开更多
Soils and waters are heavily contaminated by antimony in Xikuangshan(XKS)mine area.It is widely accepted that oxidative dissolution of sulfide minerals and aqueous dissolution are the most prevalent geochemical mechan...Soils and waters are heavily contaminated by antimony in Xikuangshan(XKS)mine area.It is widely accepted that oxidative dissolution of sulfide minerals and aqueous dissolution are the most prevalent geochemical mechanisms for the release of Sb to the environment.Bosea sp.AS-1 is an antimonite-oxidizer isolated from the mine slag in Xikuangshan Sb mine.Whole genome sequencing revealed the presence of multiple sulfur-oxidizing genes,antimony(Sb)metabolism genes and carbon fixation genes in AS-1 s genome.We therefore hypothesized that under oxic conditions,AS-1 could mediate the oxidation of sulfide and Sb(Ⅲ)in stibnite(Sb_(2)S_(3))and lead to the release of Sb.Indeed,strain AS-1 was discovered as an autotrophic Sb(Ⅲ)-oxidizer.Antimony mobilization studies conducted with strain AS-1showed significantly enhanced mobilization of Sb,and complete oxidation of released Sb and sulfur to Sb(V)and sulfate.In addition,AS-1 induced a faster release of Sb under heterotrophic condition,and new acicular minerals might form.These findings support the hypothesis that microorganisms play an important role in the mobilization and transformation of Sb in XKS mine area and may contribute to our further understanding of the Sb biogeochemical redox cycle in natural environment.展开更多
基金supported by the National Natural Science Foundation of China(No.81470418 and No.81770230)。
文摘Morphological and functional abnormalities of vascular endothelial cells(VECs) are risk factors of ischemiareperfusion in skin flaps.Signaling pathway mediated by interleukin-1 receptor(IL-1 R) is essential to hypoxia/reoxygenation(H/R) injury of VECs.While the TIR/BB-loop mimetic(AS-1) disrupts the interaction between IL-1 R and myeloid differentiation primary-response protein 88(MyD88),its role in the VECs dysfunction under H/R is unclear.In this study,we first showed that there was an infiltration of inflammatory cells and the apoptosis of VECs by using a skin flap section from patients who received flap transplantation.We then showed that the H/R treatment induced apoptosis and loss of cell migration of endothelial cell line H926 were attenuated by AS-1.Furthermore,our data suggested that AS-1 inhibits the interaction between IL-1 R and MyD88,and subsequent phosphorylation of IκB and p38 pathway,as well as the nuclear localization of NF-κB subunit p65/p50.Thus,this study indicated that the protective role of AS-1 in H/R induced cellular injury may be due to the AS-1 mediated down-regulation of IL-1 R signaling pathway.
基金This study was supported by the National Natural Science Foundation of China(No.30571842)
文摘Objective： To test whether IL-1 RI/My088-TIR mimic AS-1 can work as a new compound that targeted at blocking MyD88- dependent signaling pathway, we investigated the physical structure and biological function of AS-1. Methods：The crystallographic structure of AS-1 was examined by 1^H nuclear magnetic resonance. The toxicity of AS-1 was measured with Methyl thiazolyl tetrazolium （MTT） assay. The effect of AS-1 on phosphorylation state of p38 MAPK and IRAK-1 was observed with Western blot. Results：The crystallographic details of AS-1 demonstrated that it was a tri-peptide sequence[（F/Y）-（V/L/I）-（P/G）] of the IL-1R I -TIR domain BBloop. No toxicity of AS-1 was shown to HEK 293A cells. The phosphorylation of p38 MAPK, induced by IL-1β significantly increased from those in the control group. AS-1 significantly reduced the phosphorylation of p38 MAPK induced by IL-1β. IL-1β increased the phosphorylation of IRAK-1 significantly, which was prevented by AS-1. Conclusion：AS-1 is a competitive mimic between IL-1R I-TIR and MyD88-TIR domain, which most likely interferes with MyD88-dependent signaling pathway.
基金supported by the National Natural Science Foundation of China(Nos.42077220 and 41877320)。
文摘Soils and waters are heavily contaminated by antimony in Xikuangshan(XKS)mine area.It is widely accepted that oxidative dissolution of sulfide minerals and aqueous dissolution are the most prevalent geochemical mechanisms for the release of Sb to the environment.Bosea sp.AS-1 is an antimonite-oxidizer isolated from the mine slag in Xikuangshan Sb mine.Whole genome sequencing revealed the presence of multiple sulfur-oxidizing genes,antimony(Sb)metabolism genes and carbon fixation genes in AS-1 s genome.We therefore hypothesized that under oxic conditions,AS-1 could mediate the oxidation of sulfide and Sb(Ⅲ)in stibnite(Sb_(2)S_(3))and lead to the release of Sb.Indeed,strain AS-1 was discovered as an autotrophic Sb(Ⅲ)-oxidizer.Antimony mobilization studies conducted with strain AS-1showed significantly enhanced mobilization of Sb,and complete oxidation of released Sb and sulfur to Sb(V)and sulfate.In addition,AS-1 induced a faster release of Sb under heterotrophic condition,and new acicular minerals might form.These findings support the hypothesis that microorganisms play an important role in the mobilization and transformation of Sb in XKS mine area and may contribute to our further understanding of the Sb biogeochemical redox cycle in natural environment.
