递增注射脂多糖诱导慢性免疫应激对仔猪生长性能的影响

本试验旨在研究递增30%注射脂多糖(LPS)诱导仔猪慢性免疫应激的效果及对仔猪生长性能、营养物质表观消化率的影响。选取16头断奶仔猪,随机分为2组(对照组、LPS组),单笼饲养。LPS组隔天静脉注射LPS,初始注射剂量80μg/kg BW,随后逐次递增30%,对照组注射相应剂量的生理盐水,试验期15 d。研究发现:仔猪注射LPS第1、5天血浆白细胞介素-1β和皮质醇含量极显著高于对照组(P0.01),第9天血浆皮质醇有高于对照组的趋势(P=0.10),整个试验期内血浆结合珠蛋白含量显著高于对照组(P0.05)。与对照组相比,注射LPS仔猪在试验的3个阶段(1~4 d、5~8 d和9~15 d)平均日增重分别降低了40.00%(P0.01)、29.52%(P0.05)和19.30%(P0.05),平均日采食量分别降低了25.09%(P=0.01)、23.15%(P0.05)和19.47%(P0.05),增重耗料比与对照组无显著差异(P0.05)。与对照组相比,注射LPS仔猪饲粮干物质表观消化率下降了5.43%(P0.05),能量表观消化率有降低的趋势(P=0.06),粗脂肪和粗蛋白质表观消化率略有降低(P0.10)。试验结果表明,递增30%注射LPS成功诱导仔猪慢性免疫应激,降低了仔猪平均日采食量和营养物质表观消化率,抑制了仔猪生长。     

内毒素对肠黏膜微血管内皮细胞内分泌一氧化氮和内皮素的影响

目的是研究内毒素（lipopolysaccharide; LPS）导致肠黏膜微血管内皮细胞损伤的作用机制。将所培养的肠黏膜微血管内皮细胞分为空白对照组和LPS组，每组按时间分为4个亚组：3h、6h、9h、12h。空白对照组加维持培养液，LPS组加1μg/ml LPS培养液，在CO2培养箱内静置培养。结果表明，一氧化氮（NO）分泌明显升高，3h后达到高峰，随后逐渐降低，而内皮素（endothelin; ET）分泌急剧升高，9h后达到高峰，随后又开始逐渐降低，但一直保持在较高的水平。所以引起了NO和ET的升高可能是LPS导致肠黏膜微血管内皮细胞的损伤机制。     

Relation of Japanese <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> pv. <Emphasis Type="Italic">vesicatoria</Emphasis> with worldwide strains revealed with three specific monoclonal antibodies

Strains of Xanthomonas campestris pv. vesicatoria Dye 1978 (Xcv), the causal agent of bacterial spot, have been classified into two groups based on their ability to hydrolyze starch. Three monoclonal antibodies (MAbs), 7AH10, 5HB3, and 4AD2, were produced immunized against the living bacteria and were specific to and could distinguish Xcv strains able or unable to hydrolyze starch (Amy⁺ or Amy^–). The MAb 7AH10, obtained against strain UPB141(Amy^–) reacted in an enzyme-linked immunosorbent assay with all the Amy^– strains (n = 19) and 1 of 11 Amy⁺ strains. Against Xcv 2625, an Amy^– unusual phenotype strain, MAb 5HB3, recognized 97% of our worldwide collection of Xcvs (n = 30). Also against that strain, the MAb 4AD2 reacted with none of the homologous Amy^– phenotypes and with 90% (n = 11) of the heterologous Amy⁺ phenotypes. For all the MAbs, cross reactions with other pathovars or species were less than 4% (n = 67). By assaying a Japanese collection of strains against the three MAbs, the Amy⁺ strains were distinguished from the Amy^– strains, and their relation with other world strains could be demonstrated. All the MAbs reacted with the lipopolysaccharide fraction of the bacterial cell wall during immunoblotting.     

Effects of deltamethrin on hematological parameters and enzymatic activity in Ancistrus multispinis (Pisces, Teleostei)

Deltamethrin (DM) is a pyrethroid insecticide widely used in Brazilian crops and in pest-control programs because of its low environmental persistence and toxicity. It has been shown to exert a wide range of effects on non-targeted organisms including fishes. The aim of this study to evaluate the effects of deltamethrin through the hematological and biochemical parameters using Ancistrus multispinis as animal model. First, blood of A. multispinis was collected by cardiac puncture in order to evaluate the basal values of the total cell counts, hemoglobin and hematocrit rates. A. multispinis showed low number leukocytes, red blood cells and hemoglobin compared to other species. The intoxication with DM (0.1 or 0.3 mg kg⁻¹) induced leukocytosis and increases the number of erythrocytes and hemoglobin levels 96 h after the injection when compared to vehicle-injected animals. The injection of lipopolysaccharide (LPS) also induced a significant leukocytosis and increased the erythrocytes number 4 h after the injection. The combination of the higher dose of DM with LPS showed an additional effect in the number of leukocytes and erythrocytes. The GST and EROD activities were also evaluated in the gills and liver, respectively, 96 h after intoxication with both doses of DM. The GST activity decreased after DM intoxication in both doses while EROD activity was not significantly different. These results suggest that A. multispinis is a valuable model for ecotoxicological studies. Also, DM can affect the metabolizing system (GST) and immunologic system of A. multispinis, a condition that can be worsened in the presence of inflammation or infection.     

大肠杆菌内毒素多克隆抗体的制备与纯化

为制备大肠杆菌内毒素多克隆抗体,应用大肠杆菌内毒素作为抗原免疫5周龄家兔,辛酸-硫酸铵法和葡聚糖凝胶层析法分离提纯抗血清,十二烷基磺酸钠-聚丙烯酰胺凝胶(SDS-PAGE)电泳方法鉴定多抗纯度,紫外分光光度计测定抗体蛋白含量,ELISA法检测抗血清效价与交叉反应;大肠杆菌内毒素抗体含量和效价分别为6.95mg/ml和1:12800;成功制备出抗大肠杆菌内毒素多克隆抗体,为大肠杆菌内毒素病诊断试剂盒的研制奠定基础。     

蒲公英提取物对LPS诱导小鼠乳腺炎的减轻效应及其机制分析

旨在研究蒲公英提取物对内毒素（LPS）诱导小鼠乳腺炎的减轻效应及其机制分析。将小鼠随机分为空白组、模型组、阳性组和蒲公英提取物高、中、低剂量组。蒲公英提取物高、中、低剂量组分别按10.0、5.0、2.5 g·kg^-1灌胃给药,连续灌胃6 d,2次·d^-1,空白组灌胃等体积生理盐水。末次给药1 h后,于小鼠乳房基部分别灌注50 μL 0.2 mg·mL^-1 LPS,建立LPS诱导的小鼠乳腺炎模型,阳性组在建模后6和12 h腹腔注射5 mg·kg^-1地塞米松。24 h后取血,分离血清,剥离乳腺组织。ELISA法测定小鼠血清肿瘤坏死因子α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的含量,髓过氧化物酶（MPO）试剂盒测定小鼠血清MPO的含量,HE染色观察病理变化,Western blot法测定小鼠乳腺中TLR4蛋白以及NF-κB信号通路和MAPKs信号通路相关蛋白的表达。结果显示,蒲公英提取物高、中剂量组对LPS诱导的乳腺炎小鼠血清中TNF-α、IL-1β和IL-6的分泌有极显著抑制作用（P<0.01）,极显著降低小鼠血清中MPO的含量（P<0.01）;蒲公英提取物高、中剂量组能改善LPS诱导的小鼠乳腺组织的病理变化,极显著下调LPS诱导的小鼠乳腺中TLR4、p-IκB、p-p65、p-p38、p-JNK、p-ERK蛋白的表达（P<0.01）。结果表明,蒲公英提取物通过调控NF-κB和MAPKs信号通路对LPS诱导的小鼠乳腺炎有明显减轻作用,为蒲公英的临床开发及应用奠定了基础。     

木麻黄青枯菌的根表吸附及根内增殖与其致病性关系

对 3个不同致病的青枯菌菌株和 7个不同抗病性的木麻黄无性系苗的研究得出 ,各菌株对寄主根表吸附量具有显著差异 ,吸附量与致病性呈负相关 :各菌株在寄主根内的增殖量差别明显 ,增殖量与致病性呈正相关。病菌脂多糖 (LPS)对幼苗根的预处理 ,可使其供体菌的吸附量减少5 8.5 %　 97.3% ,但胞外多糖 (EPS)对供体菌的吸附量无明显影响。洗去EPS和LPS的菌体较完整菌体对根表的吸附量减少 6 3.3%　 74 .2 %。而仅洗去EPS的菌体对根表的吸附有所增加 ,最高达19.1%。各菌体SDS PAGE电泳图谱显示出一定差异 ,其中强毒菌株 6 0 1B具有Rf=0 .35的特异谱带。结果表明青枯菌对木麻黄的致病性与其在寄主根表的吸附情况和在根内的快速增殖具有密切关系。LPS在此病理系统中 ,起着细菌识别因子的作用 ,EPS对LPS进行掩盖 ,二者都是病菌致病性的重要因子     

Lipopolysaccharide induces expression of collagen VI in the rat lung

The involvement of the lung during the septic systemic inflammatory response elicited by administration of lipopolysaccharide (LPS) was investigated. Eight-week-old male Sprague–Dawley rats were injected i.p. with 15 mg/kg LPS. After 24 h, the lungs were excised to evaluate the cellular responses to LPS. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) analysis revealed that type VI collagen (ColVI) was extremely upregulated during sepsis in the rat lung within the first 24 h of LPS administration. Upregulation of ColVI protein and its mRNA was demonstrated by Western blot analysis, real time PCR, and immunohistochemistry. To the best of our knowledge, this is the first report demonstrating the activation of ColVI in the rat lung at the early stage of systemic inflammation. Activation of ColVI might be involved in sepsis-mediated lung fibrosis at an early stage.     

Propofol attenuates LPS-induced tumor necrosis factor-α, interleukin-6 and nitric oxide expression in canine peripheral blood mononuclear cells possibly through down-regulation of nuclear factor (NF)-κB activation

Sepsis is a major cause of mortality in intensive care medicine. Propofol, an intravenous general anesthetic, has been suggested to have anti-inflammatory properties and able to prevent sepsis induced by Gram-positive and Gram-negative bacteria by down-regulating the gene expression of pro-inflammatory cytokines. However, propofol’s anti-inflammatory effects upon canine peripheral blood mononuclear cells (PBMCs) have not yet been clarified. Here, we isolate canine PBMCs and investigate the effects of propofol on the gene expressions of both lipopolysaccharide (LPS)-induced interleukin-6 (IL-6) and tumor necrosis factor (TNF)-α and upon the production of nitric oxide (NO). Through real-time quantitative PCR and the Griess reagent system, we found that non-cytotoxic levels of propofol significantly inhibited the release of NO and IL-6 and TNF-α gene expression in LPS-induced canine PBMCs. Western blotting revealed that LPS does significantly increase the expression of inducible NO synthase (iNOS) protein in canine PBMCs, while pretreatment with propofol significantly decreases the LPS-induced iNOS protein expression. Propofol, at concentration of 25 µM and 50 µM, also significantly inhibited the LPS-induced nuclear translocation of nuclear factor (NF)-κB p65 protein in canine PBMCs. This diminished TNF-α, IL-6 and iNOS expression, and NO production was in parallel to the respective decreased NF-κB p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol. This suggests that non-cytotoxic levels of propofol pretreatment can down-regulate LPS-induced inflammatory responses in canine PBMCs, possibly by inhibiting the nuclear translocation of the NF-κB p65 protein.     

布鲁氏菌LPS对小鼠巨噬细胞中NLRP3炎症小体转录水平的影响

为研究布鲁氏菌LPS对巨噬细胞中NLRP3炎症小体的影响,本试验提取布鲁氏菌2308、RB51和△WbkA的LPS,以不同浓度与小鼠巨噬细胞相互作用,荧光定量PCR检测其对NLRP3、ASC、Caspase1、IL1-β、IL18转录水平的影响。结果显示RB51LPS和△WbkA LPS上调NLRP3炎症小体相关基因的转录水平,且呈浓度依赖性,而浓度对2308LPS调节NLRP3炎症小体相关基因转录的作用不大;且同一浓度下,RB51LPS和2308LPS比△WbkA LPS更好的调节Caspase1、IL1-β、IL18的转录水平。