Th1/Th2型免疫反应相关基因在巨噬细胞RAW 264.7应对细粒棘球绦虫原头蚴刺激时的表达谱研究

本研究旨在解析巨噬细胞RAW264.7在应对细粒棘球绦虫原头蚴刺激时其Th1、Th2型免疫反应相关基因的差异表达规律,为进一步揭示巨噬细胞抗细粒棘球绦虫原头蚴免疫调控机制奠定理论基础.将细粒棘球绦虫原头蚴和巨噬细胞RAW264.7共培养6、24、72 h,收集RAW264.7细胞,提取总RNA,构建cDNA文库,利用R...     

人参多糖对脂多糖刺激小鼠巨噬细胞的免疫调控作用

本试验旨在研究脂多糖(LPS)刺激条件下人参多糖(GPS)对小鼠单核巨噬细胞形态及免疫功能的调节作用.采用LPS刺激小鼠巨噬细胞(RAW264.7),通过测量不同浓度(1、0.5、0.1 mg/mL)GPS对细胞形态、生物酶活性、促炎症因子分泌及TLR4/NF-κB信号通路mRNA表达量的影响来研究不同浓度的GPS对L...     

发酵金针菇多糖通过核转录因子-κB-NOD样受体家族含pyrin结构域蛋白3信号通路抑制巨噬细胞炎症反应

为探究金针菇多糖(FVP)和发酵金针菇多糖(FFVP)对小鼠单核巨噬细胞(RAW264.7)炎症反应的影响与机制,以脂多糖(LPS)构建RAW264.7炎症模型,设置CON组(正常培养基)、LPS组(正常培养基+1μg/mL LPS)、FVP组(正常培养基+1μg/mL LPS+25、50或100μg/mL FVP)和FFVP组(正常培养基+1μg/mL LPS+25、50或100μg/mL FFVP),通过测定RAW264.7的细胞活力、吞噬能力、活性氧(ROS)和一氧化氮(NO)含量以及炎症因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-18(IL-18)和肿瘤坏死因子-α(TNF-α)的含量与mRNA相对表达量,比较FVP和FFVP抑制巨噬细胞炎症反应的作用;以核转录因子-κB(NF-κB)抑制剂BAY11-7082处理RAW264.7,通过Western blot检测磷酸化核转录因子-κB抑制蛋白α(p-IκBα)、NOD样受体家族含pyrin结构域蛋白3(NLRP3)、半胱天冬蛋白酶-1(Caspase-1)和IL-1β的蛋白相对表达量,探究FVP...     

Necrostatin-1对BCG感染后小鼠巨噬细胞RAW264.7凋亡的调控

5-(1H-吲哚-3-基甲基)-3-甲基-2-硫酮-4-咪唑烷酮(Necrostatin-1,Nec-1)是一种能够特异的、有效抑制细胞程序性凋亡的小分子物质.为了探讨Nec-1对卡介苗(Bacillus Calmette-Guérin,BCG)诱导的小鼠(Mus musculus)巨噬细胞RAW264.7凋亡的调控作用,本研究采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide,MTT)比色法检测细胞存活率,Annexin V和碘化丙啶(propidine iodide,PI)双染法检测细胞凋亡率,JC-1染色法检测细胞线粒体膜电位水平,采用分光光度法检测细胞内Caspase-3的酶活性,qRT-PCR和Westemblot法检测凋亡相关基因的mRNA和蛋白表达水平.结果表明,Nec-1可提高被BCG感染巨噬细胞的存活率,降低其凋亡率,通过降低线粒体膜电位水平、上调抑凋亡基因Bcl-2的表达同时下调RIP1、RIP3和BAX基因的表达水平,从而有效降低了Caspase-3的蛋白表达量及酶活性.本研究表明Nec-1可通过提高线粒体膜电位水平、并下调促凋亡蛋白的表达量,从而抑制被BCG感染后巨噬细胞的凋亡,这将有助于进一步研究结核分枝杆菌(Mycobacterium tuberculosis)与巨噬细胞间的相互作用,对于揭示结核病的致病机理具有重要意义.     

Effect of binary combination of some plant-derived molluscicides with MGK-264 or piperonyl butoxide on the reproduction of the snail Lymnaea acuminata

The effects of sub-lethal treatments (20 and 60% of 24-h LC(50)) with plant-derived molluscicides Annona squamosa, acetogenins, Argemone mexicana seed and protopine, in combination (1 + 5) with MGK-264 (ENT 8184) or piperonyl butoxide on the reproduction of Lymnaea acuminata has been studied. The plant-derived molluscicides and their active molluscicidal components, protopine and acetogenins, in combination with ENT 8184 or piperonyl butoxide caused a significant reduction in the fecundity, hatchability and survival of young snails. Combination of A squamosa seed powder with piperonyl butoxide was very effective as it caused a complete arrest of snail fecundity within 24 h of treatment. Removal of the snails to fresh water after the 96-h treatments caused a significant recovery in the fecundity of L acuminata.     

布鲁氏菌介导的mmu-miR-671-5p的靶基因预测与功能富集性分析

为进行布鲁氏菌介导的mmu-miR-671-5p的靶基因预测,本试验利用布鲁氏菌（Brucella）感染RAW264.7细胞后,分别运用miRanda和TargetScan软件进行差异表达的mmu-miR-671-5p靶基因的预测,预测的靶基因分别有11 953和9 252个,将预测结果取交集进行韦恩（Venn）分析,重叠部分的靶基因有3 681个;利用GO与KEGG进行功能富集性分析,再利用PicTar软件进一步对mmu-miR-671-5p的靶基因进行预测,将预测结果与Venn分析结果进一步取交集,结果显示,mmu-miR-671-5p的预测靶基因有7个;实时荧光定量PCR分别验证7个预测靶基因的相对表达量发现,Tnfrsf1b与Tnip1基因相对表达量极显著降低（P<0.01）;在RAW264.7细胞中转染mmu-miR-671-5p inhibitors,分别验证7个预测靶基因的相对表达量发现,Tnfrsf1b与Tnip1基因的相对表达量极显著升高（P<0.01）;对mmu-miR-671-5p与Tnfrsf1b和Tnip1的3'非翻译区（3'UTR）结合靶位点分别进行预测,结果初步表明,mmu-miR-671-5p的靶基因为Tnfrsf1b与Tnip1,其预测结合靶位点均只有1个,分别位于Tnfrsf1b和Tnip1 3'UTR全长位置的91和305 bp。本研究结果为进一步揭示mmu-miR-671-5p在布鲁氏菌感染RAW264.7细胞过程中的功能提供科学依据。     

A sesquiterpene quinone, 5-Epi-smenospongine, promotes TNF-α production in LPS-stimulated RAW 264.7 Cells

Eight sesquiterpene quinones: ilimaquinone (1), smenospongidine (3), smenospongiarine (5), smenospongine (7), and their corresponding 5-epimers 2, 4, 6, and 8, isolated from the Palauan marine sponge Hippospongia sp., were examined regarding their effects on TNF-α production in LPS-stimulated RAW 264.7 cells. 5-Epi-smenospongine (8) promoted the production of TNF-α to a level three times greater than the control at 10 μM, but compounds 1–7 did not show apparent activity. The results suggest that the cis-decaline ring and a primary amine in the benzoquinone ring are necessary for activity. This is the first study to report the modulation of TNF-α production by a sesquiterpene quinone.     

可抵抗共谋攻击的H．264／AVC压缩域数字视频水印算法

针对H.264/AVC编码标准的新特性,提出一种可抵抗共谋攻击的压缩域鲁棒数字水印算法.对Noorkami等算法进行改进,水印的安全性是基于水印嵌入位置的随机性,通过DC系数的相关改变量产生密钥从而确定水印的嵌入位置.通过改变每个宏块的一个4×4块的非零量化系数嵌入水印.仿真试验表明,本算法在保证视频的视觉质量的前提下,水印的嵌入量有很大的提高.此外,水印提取时不需要对压缩码流完全解码,并能满足实时盲检测的要求.     

Performance Analysis of Motion Estimation Parameters Based on H. 264

H, 264 is the new standard for video coding, it can save 64.46%, 48, 80% and 38.62% bit-rate compared with that of MPEG-2, H. 263 HLP and MPEG-4 ASP respectively under the same decoded picture quality, But the high encoding efficiency is at the cost of heavily increased computational complexity. The multi-prediction macroblock mode search and rate-distortion optimization mode decision that achieve high encoding efficiency are introduced firstly. Then experiment researches are performed to analyze their effect on encoding efficiency. Results show that these two kinds of technique can only achieve slightly improvement on encoding efficiency, but computational complexity increases heavily. This provides an important reference for the parameters selection when H. 264 is used in the practical application of video coding.     

脂肪酸结合蛋白4对BCG诱导巨噬细胞自噬的调控作用

旨在探讨脂肪酸结合蛋白4（fatty acid binding protein,FABP4）在牛分枝杆菌卡介苗（BCG）感染的巨噬细胞中对脂肪酸代谢与自噬的调控作用。本研究利用小干扰RNA技术敲减小鼠巨噬细胞系RAW264.7中FABP4的表达,并结合BCG感染,采用免疫印记和免疫荧光等技术,检测了FABP4蛋白表达、脂肪酸累积、脂肪酸β氧化和自噬相关蛋白表达等指标。结果表明,BCG感染极显著上调了小鼠巨噬细胞RAW264.7中FABP4的表达（P<0.001）并促进了脂肪酸的累积。FABP4小干扰RNA（siRNA-FABP4）能显著降低BCG感染后巨噬细胞中脂肪酸含量,伴随着肉毒碱棕榈酰移位酶1A（CPT1A）的表达上调（P<0.05）,与ATP产量的提升（P<0.05）。同时,siRNA-FABP4极显著下调了自噬调控关键因子AMPK及自噬相关蛋白p-ULK1、ATG5、ATG7、ATG12和LC3B的表达（P<0.01）。以上研究结果表明,在BCG感染RAW264.7细胞过程中,下调了FABP4的表达,促进了脂肪酸的氧化,减少了巨噬细胞内脂肪酸的含量,并通过AMPK信号通路抑制了细胞自噬的发生。