蜂胶对细菌脂多糖刺激下奶牛乳腺上皮细胞炎症相关基因mRNA转录水平和紧密连接蛋白的影响

本试验旨在研究中国蜂胶乙醇提取物（ethanol extract of Chinese propolis，EECP）对细菌脂多糖（lipopolysaccharide，LPS）刺激下体外培养奶牛乳腺上皮细胞炎症相关基因mRNA转录水平和紧密连接渗透性的影响。EECP中总酚酸和总黄酮含量测定采用福林酚法和硝酸铝法，并建立LPS诱导奶牛乳腺上皮细胞（bovine mammary epithelial cells，MAC-T）炎症模型，采用CCK-8法测定EECP对MAC-T相对增殖率的影响，利用实时荧光定量PCR（RT-qPCR）评估EECP对LPS诱导的MAC-T细胞炎症相关因子（IL-6、IL-8、TNF-α和IL-1β）相对mRNA转录水平；以及对紧密连接蛋白（occludin、ZO-1）相对mRNA转录水平进行检测，并进一步利用免疫荧光技术对紧密连接膜蛋白进行定位，确定EECP对LPS诱导MAC-T细胞炎症紧密连接渗透性的影响。结果显示：EECP中总酚酸含量为106.35 mg没食子酸当量（GAE）·g^-1、总黄酮含量为320.85 mg芦丁当量（RE）·g^-1；CCK-8结果显示EECP的安全浓度为0~15 μg·mL^-1，并可有效提高LPS刺激下MAC-T的活力；LPS刺激显著增加了细胞炎症相关因子IL-6、IL-8、TNF-α和IL-1β mRNA的转录量（P<0.001）；但2.5~15.0 μg·mL^-1 EECP预处理显著降低了IL-6、IL-8、TNF-α和IL-1β mRNA的转录量；与此类似，LPS刺激显著抑制了紧密连接蛋白基因（occludin、ZO-1）mRNA的转录量（P<0.01），而EECP预处理后紧密连接蛋白基因（occludin和ZO-1）mRNA的转录量显著增加（P<0.05）；免疫荧光染色试验也证实EECP能通过上调紧密连接蛋白（occludin、ZO-1）的表达，缓解LPS诱导的乳腺上皮细胞屏障功能紊乱。该结果证实，EECP对细菌脂多糖诱导奶牛乳腺上皮细胞炎症具有良好的保护作用，这为利用中国蜂胶预防奶牛乳腺炎提供了试验基础。     

6种喷雾助剂对3种药剂表面张力与接触角的影响

为达到农药减施增效的目的,将6种喷雾助剂(强力源、柔水通、有机硅、阿普顿、怀农特、倍创)与3种药剂(70%吡虫啉水分散粒剂、5%阿维菌素乳油、1.3%苦参碱水剂)混配,测定混配后药液的表面张力和接触角变化,分析药液表面张力和接触角的关系。结果表明,添加不同质量浓度的6种喷雾助剂均能降低药液的表面张力,其中,有机硅、强力源、怀农特降低3种药剂表面张力的效果最好,柔水通、阿普顿次之,倍创效果最差。有机硅、柔水通、强力源、怀农特降低3种药剂接触角的效果最好,阿普顿次之,倍创效果最差。分析了3种药剂在添加不同质量浓度的6种喷雾助剂后药液的表面张力与接触角余弦值的关系,结果显示,二者呈显著(P<0.05)负相关关系。综上,合理选用喷雾助剂有助于提高农药的利用率。研究结果可为田间施药和进一步研发喷雾助剂提供数据支持。     

Assessment of immunogenicity and protective efficacy of Microsporum canis secreted components coupled to monophosphoryl lipid-A adjuvant in a vaccine study using guinea pigs

Microsporum canis is the most common dermatophyte in pets and is of zoonotic importance but currently there is no effective vaccine available to prevent dermatophytosis. The aim of this work was to assess the immunogenicity and protective efficacy of secreted components (SC) from M. canis adjuvanted with the monophosphoryl lipid-A (MPLA), in a vaccine study using the guinea pig as an experimental model. Animals were vaccinated with either the SC adjuvanted with the MPLA, the MPLA adjuvant alone or PBS three times at two-week intervals, until 42 days prior to M. canis infection. A blind evaluation of dermatophytosis symptoms development and fungal persistence in skin was monitored weekly. The antibody response towards the SC and the levels of Interferon (IFN)γ and Interleukin-4 expressed in peripheral blood mononuclear cells were assessed along or at the end of the study period respectively. The animals that received MPLA had a significantly lower clinical score than those inoculated with PBS. However, no significant difference was observed between the guinea pigs vaccinated with the SC adjuvanted with the MPLA and those having received MPLA alone. The results also showed that vaccination induced a strong antibody response towards the SC and an increase in IFNγ mRNA level. Our results show that the MPLA adjuvant used in this vaccine study can induce per se a partial protection against a M. canis infection. Although they induce a delayed-type hypersensitivity reaction in guinea pigs, the SC do not confer a protection under the present experimental conditions.     

卵磷脂桶混助剂的特性及其在防治稻飞虱中对氟啶虫胺腈的协同增效作用

为明确新型卵磷脂类桶混助剂“融透”的特性，采用透射电镜和激光粒度分析仪表征了其物理性质，并通过室内生物测定和田间药效试验测定了其在防治稻飞虱中对氟啶虫胺腈的协同增效作用。结果表明，“融透”中含有圆形脂质体，Z平均粒径为129.5 nm。室内生物测定结果表明，“融透”可使氟啶虫胺腈对褐飞虱的毒力增加1.67倍。田间药效试验表明，“融透”可使氟啶虫胺腈对稻飞虱的防效提高9.04%~41.77%；当氟啶虫胺腈减量40%时，添加“融透”的处理对稻飞虱的防效与未添加“融透”的常量组相当。研究结果表明，卵磷脂桶混助剂“融透”可以通过形成脂质体提高药剂的利用率，从而达到增效的作用。     

无人机施药对棉蚜的防治效果及经济效益分析

为探讨采用无人机防治新疆南部地区棉蚜的可行性并筛选防治效果好的杀虫剂及具有增效作用的杀虫剂助剂，研究了采用大疆MG-1S型无人机喷施不同的杀虫剂+助剂对棉蚜的防治效果，并对无人机飞防的经济效益进行了分析。结果表明:采用MG-1S型无人机喷施22%氟啶虫胺腈悬浮剂(SC) 225 mL/hm2(制剂量，下同)时对棉蚜的防治效果最好，优于喷施70%吡虫啉水分散粒剂(WG) 30 g/hm2和70%啶虫咪WG 60 g/hm2。在添加助剂方面，以分别在70%啶虫脒WG 60g/hm2和70%吡虫啉WG 30g/hm2中添加15 mL/hm2的高工效通用助剂的防治效果最好，其次为聚合物类助剂和飞防增效剂；而在22%氟啶虫胺腈SC(150 mL/hm2)中，以添加150 mL/hm2的飞防增效剂的效果最好，其次为矿物源类、有机硅类和聚合物类助剂。采用无人机施药，在施药后3 d和5 d的防治效果与采用机械+人工拖管施药相比均无显著性差异，但无人机田间作业施药液量减少1/3，工作效率提高3倍，机械作业费减少60元/hm2，可满足现代农业高效、节药和降低成本的需要。     

细菌鞭毛蛋白作为佐剂的研究进展

细菌鞭毛蛋白作为Toll样受体5(TLR5)或NOD样受体C4(NLRC4)的配体,其结构决定了其既有抗原性又具有佐剂效应。将细菌鞭毛蛋白与外源抗原混合或融合表达,已获得多种有效的候选疫苗。细菌鞭毛蛋白佐剂效应主要是通过TLR5和NLRC4信号途径协同实现的。TLR5位于细胞表面,可触发炎性因子、趋化因子和Ⅰ型干扰素的分泌,启动天然免疫应答。NLRC4是细胞质中的模式识别受体,可识别细胞质中的多种配体并诱导相应免疫应答。另外,细菌鞭毛蛋白能募集T淋巴细胞和B淋巴细胞至次级淋巴器官,促进DCs和T淋巴细胞的活化。细菌鞭毛蛋白的可塑性将会使得以细菌鞭毛蛋白为基础的疫苗成为当前和未来研究的热点。     

新型国产白油佐剂与进口Marcol-52白油佐剂配制疫苗的比较试验

The Newcastle disease inactivated vaccines were prepared with oil adjuvants respectively from domestic and import.The experiments were done in order to compare the new domestic oil and Marcol-52 according to a series of testing indexes. The results showed that the quality of the new domestic oil was better than Marcol-52, except the absorptive quality.     

Preparation of Adjuvant Diluent for Live Vaccine against Classical Swine Fever and Evaluation of its Immune Effects

The aim of this study was to develop a novel oil-in-water (O/W) emulsion as adjuvant diluents (AD) for live vaccine against classical swine fever (CSF) that could effectively enhance the immune effect of vaccine.The AD was prepared by high-pressure homogenization technique.Formulations and preparation parameters were optimized with response surface design.Its stability, particle size, polydispersity (PDI) and Zeta potential were characterized.The humoral immune response and cellular immune response of the AD were evaluated with BALB/c mice by intramuscular injection.The particle size of the AD prepared by optimized formulation and parameters was 100.4 nm, PDI was 0.147, and Zeta potential was —28.7 mV.The experiment results showed that the AD had good stability.The AD was inoculated combined with live vaccine against CSF into BALB/c mice by intramuscular injection.The results showed that the live vaccine against CSF specific immune responses could be evoked in mice by co-inoculation with AD and vaccine.The cellular immune response levels in co-inoculated groups were significantly higher than control group (P<0.05), with obvious phenomena of higher levels of IFN-γ, IL-6 and IL-4 in serum.The result revealed that cellular immune capability significantly improved with the AD.The results strongly revealed that cellular immune capability significantly improved by introducing AD for effective immune-adjuvant for live vaccine against CSF.     

Research Progress on Cytokine Adjuvants

As an adjuvant, cytokine plays a key role in regulating the innate and adaptive immunity and enhancing the vaccine specific immune responses. The researches on the interleukin, interferon, granulocyte macrophage colony stimulating factors, tumor necrosis factors, and chemotactic factor have made considerable progress. Now we reviewed the cytokine adjuvant application in order to provide reference for the future application.