Regulation of phagosome maturation by signals from toll-like receptors

In higher metazoans, phagocytosis is essential in host defense against microbial pathogens and in clearance of apoptotic cells. Both microbial and apoptotic cells are delivered on a common route from phagosomes to lysosomes for degradation. Here, we found that activation of the Toll-like receptor (TLR) signaling pathway by bacteria, but not apoptotic cells, regulated phagocytosis at multiple steps including internalization and phagosome maturation. Phagocytosis of bacteria was impaired in the absence of TLR signaling. Two modes of phagosome maturation were observed, constitutive and inducible; their differential engagement depended on the ability of the cargo to trigger TLR signaling.     

哺乳动物抗菌肽的生物信息学预测与分析

抗菌肽(Antimicrobial peptides,AMPs)是能够抵抗病源微生物的小分子多肽,广泛分布于动植物、细菌、真菌等各种生物中,其在天然免疫中起着重要的作用。利用生物信息学方法,结合两大生物信息学门户网站ExPASy和NCBI,对截至2013年4月27日NCBI上所有的哺乳动物抗菌肽条目的上述性质进行了详细分类,并对其氨基酸长度、平均疏水性、跨膜结构、信号肽、亚细胞定位等特征进行预测及分析。     

苯酚-硫酸法测定葡萄多糖含量

用苯酚-硫酸法测葡萄多糖含量，研究显色温度、显色时间、苯酚及硫酸用量对葡萄多糖含量测定的影响。以此为基础，采用正交试验法确定最佳反应条件为：显色温度为沸水，显色时间为15min，苯酚用量为1．0ml，硫酸用量为8．0ml。     

猪细小病毒感染PK-15细胞后TLRs转录时相的变化

研究猪细小病毒(Porcine Parvovirus,PPV)感染猪肾传代细胞(PK-15 cells)后Toll样受体(Toll-like receptors,TLRs)表达水平变化情况,为探明PPV感染机制提供理论基础。试验利用荧光定量PCR方法检测PPV感染PK-15细胞后TLR1-10的转录时相变化。结果显示,PPV感染PK-15细胞后,TLR1在PPV感染后3h表达上调,约为对照组细胞的3.1倍,其他时段均低于正常水平;TLR3和TLR7 mRNA表达水平在病毒感染早期均低于正常水平;TLR4和TLR8的mRNA含量均在感染后3、12、36 h表达上调,TLR10分别在在3h和36 h表达上调,其他时段均不同程度的降低;而TLR2和TLR9 mRNA表达水平在感染后24 h开始升高,并于48 h达到峰值,分别为对照水平的10.1倍和26.7倍。研究表明,PPV感染PK-15细胞后能够诱导细胞上TLR2和TLR9在mRNA水平显著上调,PPV感染可能通过TLR2和TLR9受体介导感染。     

Ethanol embryotoxicity: direct effects on mammalian embryos in vitro

Exposure to ethanol retards growth and differentiation in cultured rat embryos during organogenesis. The development of untreated embryos is indistinguishable from growth in utero. These data suggest that the hypoplastic features of children born to chronically alcoholic mothers are due, at least in part, to a direct action of ethanol, which causes reduced embryonic cellular proliferation early in gestation.     

玫瑰露抑菌效果研究

研究了玫瑰露对4种细菌(沙门氏菌、大肠杆菌、枯草芽孢杆菌和苏云金杆菌)、3种霉菌(木霉、青霉、黑曲霉)以及2种酵母菌(啤酒酵母、葡萄酒酵母)的抑菌活性和最低抑菌浓度(MIC),采用滤纸片法测定抑菌圈、液体培养法测定最低抑菌浓度。结果表明,玫瑰露对供试细菌,不论是G+菌还是G-菌,都有较强的抑菌活性,而对霉菌、酵母菌的抑菌活性不明显;对细菌的抑菌效果以沙门氏菌最强,MIC为0.98 mL/L。说明玫瑰露既可以加入到食品中作为香料,同时可配合其他天然香料使用,组成复配防腐剂。     

An agonist of toll-like receptor 5 has radioprotective activity in mouse and primate models

The toxicity of ionizing radiation is associated with massive apoptosis in radiosensitive organs. Here, we investigate whether a drug that activates a signaling mechanism used by tumor cells to suppress apoptosis can protect healthy cells from the harmful effects of radiation. We studied CBLB502, a polypeptide drug derived from Salmonella flagellin that binds to Toll-like receptor 5 (TLR5) and activates nuclear factor-kappaB signaling. A single injection of CBLB502 before lethal total-body irradiation protected mice from both gastrointestinal and hematopoietic acute radiation syndromes and resulted in improved survival. CBLB502 injected after irradiation also enhanced survival, but at lower radiation doses. It is noteworthy that the drug did not decrease tumor radiosensitivity in mouse models. CBLB502 also showed radioprotective activity in lethally irradiated rhesus monkeys. Thus, TLR5 agonists could potentially improve the therapeutic index of cancer radiotherapy and serve as biological protectants in radiation emergencies.     

牛乳铁蛋白素-马盖宁杂合抗菌肽的设计、合成及抑菌活性

根据抗菌肽牛乳铁蛋白素(Lfcin B)与马盖宁(Magainin)的核心功能序列,将Lfcin B(1-15)和Magainin(1-12)二者连接起来,设计一种新型杂合抗菌肽LFB15-MA12,采用固相化学合成法合成获得纯度大于95%的LFB15-MA12.微量肉汤稀释法测得LFB15-MA12对金黄色葡萄球菌A...     

Induction of Zugunruhe by photostimulation of encephalic receptors in white-crowned sparrows

Daily 20-hour encephalic photophases (DEPP), transmitted (hours 0 to 20) via chronically implanted light-conducting fibers to selected sites in the basal hypothalamus of male white-crowned sparrows, were superimposed on daily 8-hour (hours 0 to 8) external ambient photophases (DAPP). Initially the birds displayed motor activity only during the 8-hour DAPP. After a delay of at least 2 weeks, some of the birds became intensively active during hours 8 to 20. We postulate that this period of "nocturnal" activity is equivalent to the nocturnal Zugunruhe shown by caged individuals of many nocturnally migratory species subjected to long days; such activity is generally regarded as the expression of migratory behavior.     

Effect of myasthenic immunoglobulin on acetylcholine receptors of intact mammalian neuromuscular junctions

Degradation of acetylcholine receptors of intact mouse neuromuscular junctions was determined in vivo and in vitro by the release of radioactivity from mouse diaphragms labeled with 125I-alpha-bungarotoxin. Treatment of mice with immunoglobulin from myasthenic patients accelerated the degradation rate to approximately three times normal, in both intact animals and organ cultures. The released radioactivity was in the form of [125I]tyrosine, confirming the nature of the degradative process. Accelerated degradation of acetylcholine receptors at neuromuscular junctions may represent an important antibody-mediated mechanisms in myasthenia gravis.     

小鼠Toll样受体9基因的克隆、表达及其活性的初步鉴定

从小鼠巨噬细胞中提取细胞总RNA,采用RT-PCR技术扩增小鼠Toll样受体9(mTLR9)cDNA,构建真核表达质粒p3XFLAG-CMV-7.1-mTLR9,转染293T细胞,利用Western Blotting检测蛋白表达,并用双荧光素酶报告基因系统检测mTLR9对其介导的信号通路下游转录因子NF-κB转录活性影响.结果表明,本试验成功克隆到小鼠TLR9cDNA,构建的重组体转染细胞后表达的蛋白分子质量与预计相符,并能激活下游转录因子NF-κB的转录活性.     

褐藻胶寡糖制备及抑菌活性研究

以平均聚合度10以下的寡糖为目标产物,将褐藻胶经不同浓度过氧化氢,在不同温度和反应时间下降解,筛选出最佳降解条件,以此降解条件下制备的寡糖对7种常见细菌和真菌进行抑菌实验.结果表明:过氧化氢体积百分比浓度3%、温度80℃、反应1.5～2 h为最佳降解条件;寡糖对细菌的生长有明显的抑制作用,最低抑菌浓度金黄色葡萄球菌、大肠杆菌及藤黄微球菌为0.625%、枯草芽孢杆菌为2.5%,寡糖对真菌抑制作用不显著.     

一株海洋放线菌的分类鉴定及抗菌活性研究

从连云港海域潮间带采集的样品中筛选得到1株产广谱、高活性抗菌物质的放线菌GB-2.该菌对藤黄微球菌、蜡样芽孢杆菌、金黄色葡萄球菌等5种革兰氏阳性菌,大肠杆菌、荧光假单胞菌等4种革兰氏阴性菌及扩展青霉、黄曲霉、点青霉、犁头霉、番茄灰霉、小麦赤霉病菌、香蕉炭疽病菌、棉花枯萎病菌、稻瘟病菌等18种真菌有显著拮抗作用.根据其形态特征、培养特征、生理生化特性和16S rRNA序列分析结果,确定其为弗氏链霉菌变种.菌株GB-2所产抗菌物质的稳定性研究表明,该物质在121 ℃ pH 1和pH 12条件下抑菌活性均不变;紫外线照射不影响其抗菌活性.结果揭示该菌在生防、食品及医药方面有潜在的应用价值.     

草酸对黄瓜过氧化物酶活性的诱导

草酸对过氧化物酶（POD）活性没有直接影响，但草酸能诱导局部叶和系统叶中不同形态的POD活性增加，且不同部位活性增加的大小和时间有明显的差异。     

Decreased RNA polymerase activity in mammalian zinc deficiency

The activity of DNA-dependent RNA polymerase has been measured in liver nuclei from suckling rats nursed by zinc-deficient dams, or by controls that were either pair-fed or given free access to the diet. In the zinc-deficient pups, the activity of the enzyme did not increase; it fell after the tenth day of life.     

不同因子对翅果油树愈伤组织诱导及分化的影响

翅果油树的繁殖和保存是目前急需解决的问题。本试验选用试管芽苗上的叶片作为外植体, 研究不同因子对愈伤组织诱导及其分化的影响。结果表明: MS+BA1mg/L+2 .4 D1 5mg/L为最佳愈伤组织诱导培养基; 在愈伤组织诱导时, 光照条件以前期暗培养、后期光培养为宜, 温度以24℃~26℃为宜; 在诱导愈伤组织芽分化时,芽分化的培养基以MS+BA1mg/L+IBA0. 05mg/L最为合适, 温度以26℃左右为宜。     

鬼针草抑菌物质提取及抑菌活性研究

采用正交试验法优化鬼针草抑菌活性成分的提取条件并研究其抑菌活性.结果表明,抑菌活性成分提取的最佳条件:95%乙醇在70℃下提取1次(2 h).鬼针草醇提物抑菌谱较宽,对金黄色葡萄球菌、枯草芽孢杆菌、水稻纹枯和荔枝疫霉的抑制效果较好,对大肠杆菌、稻瘟菌无抑制效果.对金黄色葡萄球菌、枯草芽孢杆菌的MIC分别为12.5、6.25 mg·m L-1,对水稻纹枯、荔枝疫霉的抑菌率为60.4%、31.6%.鬼针草醇提物的抑菌活性具有较好的热稳定性,对紫外光照射稳定,p H 4-8时抑菌效果稳定.     

杂合抗菌肽Scy-Hep3在毕赤酵母中的分泌表达及其抗菌活性

为获得体外稳定表达的高抗菌活性新型抗菌肽,通过重叠延伸PCR技术获得了拟穴青蟹抗菌肽Scygonadin (简记为Scy)和斜带石斑鱼抗菌肽Hepcidin 3 (简记为Hep3)的杂合抗菌肽基因scygonadinhepcidin3 (简记为scy-hep3),将scy-hep3与毕赤酵母Pichia pastoris的p PIC9K质粒连接,构建了分泌型表达载体p PIC9K-scy-hep3,用电转化法将其转化至毕赤酵母GS115菌中,经甲醇诱导表达和亲和纯化后获得杂合抗菌肽Scy-Hep3,并进行抗菌活性分析。结果表明:本研究中成功构建了重组毕赤酵母菌株GS115/p PIC9K-scy-hep3,用体积分数为0. 5%的甲醇诱导表达24 h,分泌表达上清液中获得了稳定表达的相对分子质量约24 000的表达产物,与预期杂合抗菌肽Scy-Hep3的大小相符;抗菌活性结果表明,杂合抗菌肽Scy-Hep3具有广谱抗菌活性,对被测的3种革兰氏阳性菌(溶壁微球菌Micrococcus lysodeikticus、谷氨酸棒杆菌Corynebacterium glutamicum和金黄色葡萄球菌Staphylococcus aureus)和4种革兰氏阴性菌(施氏假单胞菌Pseudomonas stutzeri、荧光假单胞菌Pseudomonas fluorescens、嗜水气单胞菌Aeromonas hydrophila和弗氏志贺氏菌Shigella flexneri)具有较强的抗菌活性(最小抑菌浓度MIC<12μmol/L);对比于单一的抗菌肽Scy和Hep3,杂合抗菌肽Scy-Hep3的抗菌活性显著增强。本研究结果将为杂合抗菌肽Scy-Hep3的生产和开发应用提供数据资料。