Streptococcus equi ssp. zooepidemicus (S. zooepidemicus) is a zoonotic pathogen with worldwide distribution. Lacking suitable vaccine and virulent maker is still bottleneck to control this infection. An immunoproteo...Streptococcus equi ssp. zooepidemicus (S. zooepidemicus) is a zoonotic pathogen with worldwide distribution. Lacking suitable vaccine and virulent maker is still bottleneck to control this infection. An immunoproteomic approach has been used to screen the membrane-associated and cell wall-associated proteins of S. zooepidemicus isolate in China CY to discover vaccine candidate antigens and therapeutic agents. Finally, 11 membrane-associated proteins, and 13 cell wall-associated proteins were successfully identified. BLAST (www.sanger.ac.uk) results also indicated that nucleotide sequences of majority identified proteins shared high homology (60%) with S. zooepidemicus, except for AC1-3, AC5, AC8, and AC13. Moreover, genes for 7 of the identified proteins were detected from CY; compared with ST171, 3 of them (AM1, AM8 and AC11) were only found in virulent strains (CY). All of the proteins identified in this study remain not to be reported in S. zooepidemicus. Some of the proteins serve a vital role in the immune system and reproduction of host species according to available data, while the functions of the rest were seldom researched.展开更多
Deep-sowing is an important method for avoiding drought stress in crop species,including maize.Identifying candidate genes is the groundwork for investigating the molecular mechanism underlying maize deep-sowing toler...Deep-sowing is an important method for avoiding drought stress in crop species,including maize.Identifying candidate genes is the groundwork for investigating the molecular mechanism underlying maize deep-sowing tolerance.This study evaluated four traits(mesocotyl length at 10 and 20 cm planting depths and seedling emergence rate on days 6 and 12)related to deep-sowing tolerance using a large maize population containing 386 inbred lines genotyped with 0.5 million high-quality single nucleotide polymorphisms(SNPs).The genomewide association study detected that 273 SNPs were in linkage disequilibrium(LD)with the genetic basis of maize deep-sowing tolerance.The RNA-sequencing analysis identified 1944 and 2098 differentially expressed genes(DEGs)in two comparisons,which shared 281 DEGs.By comparing the genomic locations of the 273 SNPs with those of the 281 DEGs,we identified seven candidate genes,of which GRMZM2G119769 encoded a sucrose non-fermenting 1 kinase interactor-like protein.GRMZM2G119769 was selected as the candidate gene because its homologs in other plants were related to organ length,auxin,or light response.Candidate gene association mapping revealed that natural variations in GRMZM2G119769 were related to phenotypic variations in maize mesocotyl length.Gene expression of GRMZM2G119769 was higher in deep-sowing tolerant inbred lines.These results suggest that GRMZM2G119769 is the most likely candidate gene.This study provides information on the deep-sowing tolerance of maize germplasms and identifies candidate genes,which would be useful for further research on maize deep-sowing tolerance.展开更多
Our previous studies demonstrated that JWA, a novel retinoic acids responsive and cytoskeleton related gene, is associated with cell differentiation and apoptosis. In the present study, to elucidate if the JWA is a no...Our previous studies demonstrated that JWA, a novel retinoic acids responsive and cytoskeleton related gene, is associated with cell differentiation and apoptosis. In the present study, to elucidate if the JWA is a novel kind of microtubule-associated proteins (MAPs) and functionally link to microtubule, we first successfully identified JWA from the physically purified MAPs complex of rat brain tissues. The results of co-immunoprecipitation, gene trans-fection and immunofluorescence microscopy assays from HBE and NIH3T3 cells provide strong evidence for a linkage between JWA and b-tubulin. In general, JWA is stably binding to b-tubulin whenever microtubule is polymerized or not, and it may be critical to the mitosis process. In addi-tion, by use of the antisense oligonucleotides technique, we also showed that JWA is a negative modulator on intracellu-lar amino acids in PC12 cells. Further analysis indicated that JWA selectively regulates both taurine, an inhibitory amino acid, and glutamate, an excitatory amino acid. In conclusion, JWA is not only structurally associated, but also a novel functional MAP.展开更多
目的研究重组人粒细胞集落刺激因子(Reeomhinant human gmnulocyte colony-stimulating factor,rhG-CSF)对脑梗死周围区的微管相关蛋白(Microtubule-associated protein 2,MAP-2)表达的影响。方法用线栓法建立大鼠大脑中动脉缺血再灌注...目的研究重组人粒细胞集落刺激因子(Reeomhinant human gmnulocyte colony-stimulating factor,rhG-CSF)对脑梗死周围区的微管相关蛋白(Microtubule-associated protein 2,MAP-2)表达的影响。方法用线栓法建立大鼠大脑中动脉缺血再灌注模型,观察rhG-CSF对神经功能缺损评分的影响;应用半定量逆转录一聚合酶链式反应(RT-PCR)方法和免疫组织化学方法分别测定梗死周围区MAP-2的mRNA和蛋白表达水平的变化。结果与对照组相比,治疗组的神经功能缺损评分较低,脑缺血周围区MAP-2的mRNA和蛋白表达增高。结论rhG-CSF可以促进脑缺血周围区的微管相关蛋白的表达,从而减轻脑缺血再灌注损伤。展开更多
多系统萎缩(multiple system atrophy,MSA)是一类神经系统退行性疾病,其病理特征是胶质细胞中出现含有不溶性α突触核蛋白(α-synuclein)的胞质包涵体.研究显示,α-synuclein在多系统萎缩的发病机制中有重要作用,但其毒性的分子机制目...多系统萎缩(multiple system atrophy,MSA)是一类神经系统退行性疾病,其病理特征是胶质细胞中出现含有不溶性α突触核蛋白(α-synuclein)的胞质包涵体.研究显示,α-synuclein在多系统萎缩的发病机制中有重要作用,但其毒性的分子机制目前还不清楚.本文在前期研究氧化应激条件下α-synuclein引起细胞内钙稳态失衡,提出了以氧化应激为连接的多系统萎缩中,胶质细胞死亡的新假说的基础上,深入分析了α-synuclein过表达导致U251细胞变性死亡的分子机制.首先证明过表达α-synuclein的U251细胞出现生长速度减慢、氧化应激水平增加和钙离子瞬时受体电位通道蛋白(transient receptor potential channel-1,TRPC1)表达量升高,而且细胞存活率的变化可通过下调TRPC1的表达得以恢复,说明TRPC1在α-synuclein过表达细胞死亡中发挥了重要作用;其次,研究发现α-synuclein稳转U251细胞中出现了明显的自噬水平增加和细胞凋亡的特征,表明α-synuclein通过作用于内质网钙泵以及细胞膜上的瞬时受体电位钙通道TRPC1,破坏了细胞内的钙稳态,进而影响自噬和凋亡,增加了U251细胞对于过氧化氢的敏感性,这可能是导致多系统萎缩病人脑内胶质细胞死亡的原因.展开更多
目的探讨细胞外信号调节激酶(ERK)抑制剂对间歇低氧大鼠海马CA1区神经细胞自噬相关蛋白表达及认知功能的影响。方法成年雄性Wistar大鼠96只,采用随机数字法分为正常对照组(UC组)、5%间歇低氧组(5%IH组)、ERK抑制剂组(U0126组),每组又分...目的探讨细胞外信号调节激酶(ERK)抑制剂对间歇低氧大鼠海马CA1区神经细胞自噬相关蛋白表达及认知功能的影响。方法成年雄性Wistar大鼠96只,采用随机数字法分为正常对照组(UC组)、5%间歇低氧组(5%IH组)、ERK抑制剂组(U0126组),每组又分为7、14、21、28 d 4个时间点,每个时间点8只大鼠。应用透射电镜观察大鼠海马CA1区神经细胞线粒体、自噬体等超微结构的改变;免疫组织化学法检测海马CA1区ERK1/2、Beclin-1、微管相关蛋白1轻链3(LC3)蛋白表达;TUNEL检测海马CA1区神经细胞凋亡情况;Morris水迷宫检测各组大鼠学习记忆功能。结果与UC组比较,5%IH组7、14、21、28 d海马CAl区神经细胞ERK1/2蛋白表达明显增加(P<0.05);与5%IH组比较,U0126组7、14、21、28 d海马CAl区神经细胞ERK1/2蛋白表达明显降低(15.48±2.00 vs 27.91±2.15,15.51±1.93 vs 35.86±2.58,15.57±2.09 vs 47.04±3.96,15.43±1.97 vs 59.03±4.09,P<0.05)。与UC组比较,5%IH组和U0126组7、14、21、28 d海马CAl区神经细胞Beclin-1和LC3蛋白表达及神经元凋亡指数明显增加(P<0.05);与5%IH组比较,U0126组7、14、21、28 d海马CAl区神经细胞Beclin-1和LC3蛋白表达及神经元凋亡指数明显降低(P<0.05)。与UC组比较,5%IH组和U0126组大鼠7、14、21、28 d逃避潜伏期明显延长,跨越目标象限时间明显缩短(P<0.05);与5%IH组比较,U0126组大鼠7、14、21、28 d逃避潜伏期明显缩短,跨越目标象限时间明显延长(P<0.05)。结论ERK抑制剂U0126通过阻断ERK信号通路抑制大鼠海马神经细胞自噬的激活,进而减轻神经细胞损伤,改善认知功能。展开更多
基金supported by the Program for New Century Excellent Talents (NCET) at the University of China (NCET-08-0794)the Key Technology R&D Program of Jiansu Province, China (BE2009388)+1 种基金the Fundamental Research Funds for the Central Universities, China (KYT 201003)the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China
文摘Streptococcus equi ssp. zooepidemicus (S. zooepidemicus) is a zoonotic pathogen with worldwide distribution. Lacking suitable vaccine and virulent maker is still bottleneck to control this infection. An immunoproteomic approach has been used to screen the membrane-associated and cell wall-associated proteins of S. zooepidemicus isolate in China CY to discover vaccine candidate antigens and therapeutic agents. Finally, 11 membrane-associated proteins, and 13 cell wall-associated proteins were successfully identified. BLAST (www.sanger.ac.uk) results also indicated that nucleotide sequences of majority identified proteins shared high homology (60%) with S. zooepidemicus, except for AC1-3, AC5, AC8, and AC13. Moreover, genes for 7 of the identified proteins were detected from CY; compared with ST171, 3 of them (AM1, AM8 and AC11) were only found in virulent strains (CY). All of the proteins identified in this study remain not to be reported in S. zooepidemicus. Some of the proteins serve a vital role in the immune system and reproduction of host species according to available data, while the functions of the rest were seldom researched.
基金supported by the National Key R&D Program of China(2018YFD0100903)the China Agriculture Research System of MOF and MARA(CARS-02-13)the Natural Science Fund of Liaoning Province,China(20170540806)。
文摘Deep-sowing is an important method for avoiding drought stress in crop species,including maize.Identifying candidate genes is the groundwork for investigating the molecular mechanism underlying maize deep-sowing tolerance.This study evaluated four traits(mesocotyl length at 10 and 20 cm planting depths and seedling emergence rate on days 6 and 12)related to deep-sowing tolerance using a large maize population containing 386 inbred lines genotyped with 0.5 million high-quality single nucleotide polymorphisms(SNPs).The genomewide association study detected that 273 SNPs were in linkage disequilibrium(LD)with the genetic basis of maize deep-sowing tolerance.The RNA-sequencing analysis identified 1944 and 2098 differentially expressed genes(DEGs)in two comparisons,which shared 281 DEGs.By comparing the genomic locations of the 273 SNPs with those of the 281 DEGs,we identified seven candidate genes,of which GRMZM2G119769 encoded a sucrose non-fermenting 1 kinase interactor-like protein.GRMZM2G119769 was selected as the candidate gene because its homologs in other plants were related to organ length,auxin,or light response.Candidate gene association mapping revealed that natural variations in GRMZM2G119769 were related to phenotypic variations in maize mesocotyl length.Gene expression of GRMZM2G119769 was higher in deep-sowing tolerant inbred lines.These results suggest that GRMZM2G119769 is the most likely candidate gene.This study provides information on the deep-sowing tolerance of maize germplasms and identifies candidate genes,which would be useful for further research on maize deep-sowing tolerance.
基金supported by the National Natural Science Foundation of China(Grant Nos.30070664 and 30170812)the Foundation of High-Tech Key Project of Educational Department of Jiangsu Province(Grant No.JH01-049)+1 种基金the National Key Basic Research and Development Project(973)(Grant No.2002CB512905)the Project of the Ministry of Science and Technology of China(Grant Nos.2001-50 and 2000-026).
文摘Our previous studies demonstrated that JWA, a novel retinoic acids responsive and cytoskeleton related gene, is associated with cell differentiation and apoptosis. In the present study, to elucidate if the JWA is a novel kind of microtubule-associated proteins (MAPs) and functionally link to microtubule, we first successfully identified JWA from the physically purified MAPs complex of rat brain tissues. The results of co-immunoprecipitation, gene trans-fection and immunofluorescence microscopy assays from HBE and NIH3T3 cells provide strong evidence for a linkage between JWA and b-tubulin. In general, JWA is stably binding to b-tubulin whenever microtubule is polymerized or not, and it may be critical to the mitosis process. In addi-tion, by use of the antisense oligonucleotides technique, we also showed that JWA is a negative modulator on intracellu-lar amino acids in PC12 cells. Further analysis indicated that JWA selectively regulates both taurine, an inhibitory amino acid, and glutamate, an excitatory amino acid. In conclusion, JWA is not only structurally associated, but also a novel functional MAP.
文摘目的研究重组人粒细胞集落刺激因子(Reeomhinant human gmnulocyte colony-stimulating factor,rhG-CSF)对脑梗死周围区的微管相关蛋白(Microtubule-associated protein 2,MAP-2)表达的影响。方法用线栓法建立大鼠大脑中动脉缺血再灌注模型,观察rhG-CSF对神经功能缺损评分的影响;应用半定量逆转录一聚合酶链式反应(RT-PCR)方法和免疫组织化学方法分别测定梗死周围区MAP-2的mRNA和蛋白表达水平的变化。结果与对照组相比,治疗组的神经功能缺损评分较低,脑缺血周围区MAP-2的mRNA和蛋白表达增高。结论rhG-CSF可以促进脑缺血周围区的微管相关蛋白的表达,从而减轻脑缺血再灌注损伤。
文摘目的探讨细胞外信号调节激酶(ERK)抑制剂对间歇低氧大鼠海马CA1区神经细胞自噬相关蛋白表达及认知功能的影响。方法成年雄性Wistar大鼠96只,采用随机数字法分为正常对照组(UC组)、5%间歇低氧组(5%IH组)、ERK抑制剂组(U0126组),每组又分为7、14、21、28 d 4个时间点,每个时间点8只大鼠。应用透射电镜观察大鼠海马CA1区神经细胞线粒体、自噬体等超微结构的改变;免疫组织化学法检测海马CA1区ERK1/2、Beclin-1、微管相关蛋白1轻链3(LC3)蛋白表达;TUNEL检测海马CA1区神经细胞凋亡情况;Morris水迷宫检测各组大鼠学习记忆功能。结果与UC组比较,5%IH组7、14、21、28 d海马CAl区神经细胞ERK1/2蛋白表达明显增加(P<0.05);与5%IH组比较,U0126组7、14、21、28 d海马CAl区神经细胞ERK1/2蛋白表达明显降低(15.48±2.00 vs 27.91±2.15,15.51±1.93 vs 35.86±2.58,15.57±2.09 vs 47.04±3.96,15.43±1.97 vs 59.03±4.09,P<0.05)。与UC组比较,5%IH组和U0126组7、14、21、28 d海马CAl区神经细胞Beclin-1和LC3蛋白表达及神经元凋亡指数明显增加(P<0.05);与5%IH组比较,U0126组7、14、21、28 d海马CAl区神经细胞Beclin-1和LC3蛋白表达及神经元凋亡指数明显降低(P<0.05)。与UC组比较,5%IH组和U0126组大鼠7、14、21、28 d逃避潜伏期明显延长,跨越目标象限时间明显缩短(P<0.05);与5%IH组比较,U0126组大鼠7、14、21、28 d逃避潜伏期明显缩短,跨越目标象限时间明显延长(P<0.05)。结论ERK抑制剂U0126通过阻断ERK信号通路抑制大鼠海马神经细胞自噬的激活,进而减轻神经细胞损伤,改善认知功能。