程序性细胞死亡因子10抑制Ⅰ型干扰素表达并促进FMDV复制     

曾宗波  马旭升  罗志宽  齐亚银  郑海学 《畜牧兽医学报》2021,52(2):450-459

旨在探究宿主蛋白程序性细胞死亡因子10（programmed cell death factor 10，PDCD10）通过抑制Ⅰ型干扰素表达进而促进口蹄疫病毒（foot-and-mouth disease virus，FMDV）的复制。首先，本研究验证了过表达和沉默PDCD10对FMDV复制的影响，接着利用双荧光素酶报告系统探究PDCD10对Ⅰ型干扰素信号通路活化的影响，最后，利用实时荧光定量PCR探究PDCD10对Ⅰ型干扰素通路下游刺激基因（IFN-stimulated genes，ISGs）转录的影响。结果表明，过表达PDCD10显著促进FMDV的复制，沉默PDCD10显著抑制FMDV的复制。与对照相比，过表达PDCD10后感染仙台病毒（Sendai virus，SeV）的细胞培养液上清液显著促进FMDV复制，进一步，PDCD10显著抑制SeV诱导的IFN-β启动子以及NF-κB的激活且呈剂量依赖性，并且PDCD10负调控Ⅰ型干扰素通路信号分子转录，最后还发现PDCD10负调控Ⅰ型干扰素下游ISGs转录。本研究结果为深入探究PDCD10在抗病毒天然免疫中的作用积累了资料。  相似文献   

鸡Ii结合Rab5a和Rab7b分子的分析     

陈芳芳  桂亚萍  于凤梅  张俊  谈阳  李锦春  刘翠艳  查丽莎 《畜牧兽医学报》2021,52(12):3588-3597

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。  相似文献   

施氮水平对7S亚基缺失大豆根系形态和结瘤固氮的影响   总被引：2，自引：0，他引：2  

姜妍  王清泉  李远明  王绍东  刘伟 《大豆科学》2017,36(2)

为有效推广功能型大豆7S亚基缺失品种,以7S亚基缺失大豆品系东富2号为研究对象,设置4种施氮水平(纯N),N0(0 mg·kg~(-1))、N1(25 mg·kg~(-1))、N2(50 mg·kg~(-1))、N3(75 mg·kg~(-1)),采用桶栽法研究大豆根系形态和结瘤固氮对不同施氮水平的响应。结果表明:N1(25 mg·kg~(-1))水平下根系干重加大,根冠比增大,根瘤固氮潜力高,单株产量较高。N2(50 mg·kg~(-1))水平下根长、根表面积、根体积在生育后期增长较快,根系干重较大,根冠比低,固氮酶活性最高,单株籽粒产量最高。N3(75 mg·kg~(-1))水平下植株干重较大,无效生长较多,根瘤数少,固氮潜力和根冠比低,单株产量不高。综合籽粒产量和根系特性指标,功能型大豆7S亚基缺失品系东富2号的适宜施肥量为25~50 mg·kg~(-1)。  相似文献   

miR-140-3p inhibits viability,invasion and migration of non-small-cell lung cancer A549 cells by targeting PD-L1     

ZHANG Yu-xuan  LI Chun-wei  MAO Wen-hao  ZHU Ke-yan  SHAO Yang-qian  DENG Xiao-ming 《园艺学报》2019,35(1):8-14

AIM: To explore the target relationship between microRNA-140-3p (miR-140-3p) and programmed cell death ligand 1 (PD-L1) and their effect on the viability, migration and invasion of non-small-cell lung cancer A549 cells.METHODS: RT-qPCR was used to detect the miR-140-3p expression in HLF-1, A549 and H1299 cells, and then the A549 cells with the most significant difference were selected as the subsequent research object. TargetScan software and dual-luciferase reporter assay were performed to predict and confirm the target relationship between miR-140-3p and PD-L1. RT-qPCR and Western blot were used to determine the effects of miR-140-3p mimic and inhibitor on PD-L1 expression level. MTT assay was used to detect the viability of A549 cells. Transwell assay was performed to detect the migration and invasion abilities of the A549 cells.RESULTS: miR-140-3p was significantly down-regulated in the A549 cells and H1299 cells (P<0.05). Transfection with miR-140-3p mimic decreased the expression of PD-L1 and inhibited the viability, migration and invasion of the A549 cells. Transfection with pcDNA3.0-PD-L1 reversed the inhibitory effect of miR-140-3p on the viability, migration and invasion of the A549 cells.CONCLUSION: miR-140-3p inhibits the viability, migration and invasion of A549 cells by targeting PD-L1.  相似文献   

内生菌与雷公藤细胞悬浮共培养的生理生化特性     

宋欢  宋萍  封磊  洪伟  吴承祯  王一惠  申露文 《热带作物学报》2017,(2):288-294

将2株雷公藤内生真菌Fusarium oxysporum NS33与Penicillium steckii NS6、2株内生细菌Enterobacter cloacae sub sp LG3与Serratia marcescens LY1及其组合分别与雷公藤细胞悬浮共培养,对不同培养体系内雷公藤细胞的生长及其生理生化特征进行研究。结果显示,在共培养前期,与对照相比,接种单一内生菌株提高了细胞的干重,其中菌株NS6的促生效果最明显;而在共培养后期,无论是单一内生菌还是混合内生菌均对细胞生长具有抑制作用。内生真菌和菌株组合处理的培养液p H值有明显的升高,而内生细菌LY1则明显降低了培养液的p H值,其具有产酸性。另外,当雷公藤细胞同混合菌株共培养时,培养基中总糖消耗量是最大的,而接种单独菌株时则对细胞可溶性蛋白含量具有一定的提高作用。接种内生菌会影响雷公藤细胞的POD、CAT及SOD活性,与对照相比,接种单独菌株会更加提升POD与CAT的活性,而细胞MDA含量则明显下降。  相似文献   

The M43 domain-containing metalloprotease RcMEP1 in Rhizoctonia cerealis is a pathogenicity factor during the fungus infection to wheat          下载免费PDF全文

PAN Li-jun  LU Lin  LIU Yu-ping  WEN Sheng-xian  ZHANG Zeng-yan 《农业科学学报》2020,19(8):2044-2055

Wheat(Triticum aestivum L.) is an important staple crop for global human. The necrotrophic fungus Rhizoctonia cerealis is the causal pathogen of sharp eyespot, a devastating disease of wheat. Herein, we identified RcMEP1, a zinc metalloproteaseencoding gene from R. cerealis genomic sequences, and characterized its pathogenesis function. RcMEP1 expressed at markedly-high levels during R. cerealis infection process to wheat. The predicted protein RcMEP1 comprises of 287 amino acid residues and contains a signal peptide and a M43 metalloprotease domain harboring the active site motif(HEVGHWLGLYH). The assays of Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana leaves indicated that RcMEP1 is an apoplastic elicitor of cell death, and that the predicted signal peptide functions and is required for secretion and cell death-induction. The purified RcMEP1 protein and its M43 domain peptide were individually able to induce plant cell death and H2 O2 accumulation, and to inhibit expression of host chitinases when infiltrated into wheat and N. benthamiana leaves, while the M43 domain-deleting peptide and negative control lacked the capacity. Moreover, compared with the control pretreatment, the purified RcMEP1 protein or its M43-domain peptide resulted in enhanced pathogenesis in the inoculated wheat, whereas the M43 domain-deleting peptide failed. These results suggest that RcMEP1 acted as an important pathogenicity factor during R. cerealis infection to wheat and that its signal peptide and M43 domain are required for the secretion and pathogenesis of RcMEP1. This study provides insights into pathogenesis role of M43 domain-containing metalloproteases during R. cerealis infection to wheat.  相似文献   

Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens          下载免费PDF全文

ZENG Xian-ying  CHEN Xiao-han  MA Shu-jie  WU Jiao-jiao  BAO Hong-mei  PAN Shu-xin  LIU Yan-jing  DENG Guo-hua  SHI Jian-zhong  CHEN Pu-cheng  JIANG Yong-ping  LI Yan-bing  HU Jing-lei  LU Tong  MAO Sheng-gang  GUO Xing-fu  LIU Jing-li  TIAN Guo-bin  CHEN Hua-lan 《农业科学学报》2020,19(9):2294-2300

We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control.  相似文献   

Zerumbone attenuates MPP⁺-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress     

CAO Rui-ping  WANG Jiao  WANG Ce 《园艺学报》2018,34(6):1061-1066

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.  相似文献   

The Safety Evaluation of Cefalonium Intramammary Infusion (Dry Cow)     

HUA WeiYi  LIU YiMing  XU Fei  LU YongQiang  KONG Mei  WANG HaiTing  HUANG HuiLi  WANG HongLei  WU LianYong  LI XiuBo 《中国农业科学》2019,52(2):359-366

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EPS364, a Novel Deep-Sea Bacterial Exopolysaccharide,Inhibits Liver Cancer Cell Growth and Adhesion     

Yun Wang  Ge Liu  Rui Liu  Maosheng Wei  Jinxiang Zhang  Chaomin Sun 《Marine drugs》2021,19(3)

The prognosis of liver cancer was inferior among tumors. New medicine treatments are urgently needed. In this study, a novel exopolysaccharide EPS364 was purified from Vibrio alginolyticus 364, which was isolated from a deep-sea cold seep of the South China Sea. Further research showed that EPS364 consisted of mannose, glucosamine, gluconic acid, galactosamine and arabinose with a molar ratio of 5:9:3.4:0.5:0.8. The relative molecular weight of EPS364 was 14.8 kDa. Our results further revealed that EPS364 was a β-linked and phosphorylated polysaccharide. Notably, EPS364 exhibited a significant antitumor activity, with inducing apoptosis, dissipation of the mitochondrial membrane potential (MMP) and generation of reactive oxygen species (ROS) in Huh7.5 liver cancer cells. Proteomic and quantitative real-time PCR analyses indicated that EPS364 inhibited cancer cell growth and adhesion via targeting the FGF19-FGFR4 signaling pathway. These findings suggest that EPS364 is a promising antitumor agent for pharmacotherapy.  相似文献