Comparison of flagellin and an oil-emulsion adjuvant in inactivated Newcastle disease vaccine in stimulation of immunogenic parameters

The present study was designed to investigate the potential application of native (N) and recombinant (truncated modified [tmFliC] and full-length [flFliC]) flagellin proteins along with inactivated Newcastle disease virus (NDV). Fifty six SPF chickens were immunized twice with PBS (control), inactivated NDV (Ag), inactivated NDV/flFliC (AgF), inactivated NDV/tmFliC (AgT), inactivated NDV/N (AgN), commercial vaccine containing Montanide (Vac) and Vac/N (VacN), with a two-week interval. Blood was collected weekly and spleens were harvested after chickens were sacrificed. Interleukin-6 (IL-6) and tumor necrotic factor-α (TNF-α) gene expression in peripheral blood mononuclear cells were analyzed by Real-Time PCR. Antibody response was assessed by haemagglutination inhibition (HI). Cellular activity was quantified by MTT assay. Results showed that the most IL-6 and TNF-α gene expression was observed in AgF group (P < 0.01). The lowest gene expression among vaccinated groups was observed in Ag group for IL-6 and Ag and Vac group for TNF-α. The highest HI titer was observed in Vac, VacN, AgF and AgT groups. The AgF group showed the highest cellular activity (P < 0.01). In conclusion, flagellin-adjuvanted groups showed a pro-inflammatory effect and acted similarly to or better than the Vac group. Hence, flagellin can be proposed as a potential adjuvant for ND vaccine.     

大豆GmRLP19基因克隆及胁迫应答模式分析

以大豆品种Williams82为试验材料,克隆GmRLP19基因,通过生物信息学分析qPCR检测探讨该基因不同生育期品种、不同组织、不同非生物胁迫下表达模式。结果表明,该基因编码区全长3 138 bp,编码1 045个氨基酸,编码蛋白具有胞外信号肽,LRRNT域,富含亮氨酸重复结构域,跨膜结构域和一段胞内短肽,属于受体类蛋白家族,定位于质膜。该基因在不同生育期品种间差异表达,在豆和豆荚中特异性表达,且在5种非生物胁迫中均有响应。研究为探究WRKY20基因功能及挖掘和完善基因调控通路奠定基础。     

商品化猪丹毒疫苗分类及特点

文章介绍了5种商品化猪丹毒疫苗的种类、毒株组成、抗原含量及相应免疫程序,为了解各种类型的猪丹毒商品化疫苗提供参考。     

坛紫菜叶状体的抗生素联合除菌效果

为研究坛紫菜及其外生菌群对氯霉素、竹桃霉素和头孢噻肟3种抗生素的敏感性,通过将其与氨苄青霉素、卡那霉素和庆大霉素的组合,优化坛紫菜叶状体的除菌方法,并利用涂布平板、qRT-PCR及16S rRNA测序分析其除菌效果。结果显示,当坛紫菜先以氨苄青霉素(300 mg/L)、卡那霉素(100 mg/L)和庆大霉素(100 mg/L)混合处理18 h,再以氯霉素(50 mg/L)、头孢噻肟(200 mg/L)和竹桃霉素(50 mg/L)混合处理4 d,叶状体的健康率保持在96.3%以上,而对可培养细菌的抑制率达到99.9%。根据qRT-PCR和16S rRNA的结果可知,该种组合方法较前3种抗生素的组合,其菌总数下降41.5%,微生物的丰度和多样性指数明显下降。其中,假单胞菌、交替赤杆菌、交替单胞菌以及海杆菌等得到针对性的抑制。研究表明,多种抗生素的优化组合能够对坛紫菜叶片的菌群结构产生显著影响,使其受到抑制,但仍无法达到绝对除菌的效果。     

Construction of recombinant capripoxviruses as vaccine vectors for delivering foreign antigens: Methodology and application

Goatpox (GTP), sheeppox (SPP) and lumpy skin disease (LSD) are three severe diseases of goat, sheep and cattle. Their typical clinical symptoms are characterized by vesicles, papules, nodules, pustules and scabs on animal skins. The GTP, SPP and LSD are caused by goatpox virus (GTPV), sheeppox virus (SPPV) and lumpy skin disease virus (LSDV), respectively, all of which belong to the genus Capripoxvirus in the family Poxviridae. Several capripoxvirus (CaPV) isolates have been virulently attenuated through serial passaging in vitro for production of live vaccines. CaPV-based vector systems have been broadly used to construct recombinant vaccines for delivering foreign antigens, many of which have been demonstrated to induce effective immune protections. Homologous recombination is the most commonly used method for constructing recombinant CaPVs. Here, we described a methodology for generation of recombinant CaPVs by the homologous recombination, and further reviewed CaPV-vectored vaccines for delivering foreign antigens.     

H2S的信号分子作用及其对果蔬采后生理代谢的调控研究进展

硫化氢（H2S）作为气体信号分子，参与调控植物体的一系列生命活动，并在果蔬采后生理代谢过程中发挥着重要作用。本文综述了H2S的理化性质、发展应用历史及其在果蔬采后生理调控机制中的信号分子作用，讨论了H2S对果蔬生理代谢的调控作用，包括对果蔬采后抗逆性和品质的影响，以期为采后果蔬的贮藏保鲜提供理论参考。     

河北省长针线虫新纪录种松长针线虫Longidorus pinus Xu,Ye,Wang&Zhao的形态与分子鉴定

2018 年，在河北省保定市园林植物紫荆（Cercis chinensis）根围分离到一种长针线虫。经形态 学观察和 28S rDNA D2D3 区序列分析，将其鉴定为松长针线虫（Longidorus pinus Xu, Ye, Wang & Zhao）。 其雌虫主要形态特征为：雌虫体长 3 048~3 464 μm，唇区明显缢缩，宽 9.5~10.5 μm，侧器囊袋状，齿尖 针长 66.0~69.5 μm，导环距体前端 27.5~30.0 μm，尾长 31~33 μm，短圆锥形，尾长与肛门处体宽比值＝ 1.5~1.6。松长针线虫河北种群 28S rDNA D2D3 区与 GenBank 数据库的山西种群进行序列比对，相似性为 99.2%~99.9%。松长针线虫是河北省长针线虫新纪录种。紫荆是松长针线虫的新寄主。     

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

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.     

甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物纳米微球作为口蹄疫病毒核酸疫苗递送载体的特性评价

旨在对制备的甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物[poly （D，L-lactide-co-glycolide），PLGA]纳米微球作为口蹄疫病毒（foot-and-mouth disease virus，FMDV）核酸疫苗递送载体进行评价。采用西佛碱反应和元素分析制备具有一定取代度的甘露糖修饰的壳聚糖衍生物（mannose modified chitosan，MCS），然后，经双重乳化挥发法制备得到甘露糖修饰的壳聚糖PLGA纳米微球（MCS-PLGA-NPs）。采用纳米粒径仪检测MCS-PLGA-NPs粒径分布和表面电势（zeta）、扫描电镜考察其形态、琼脂糖凝胶电泳观察其对质粒的吸附和吸附质粒后抵抗核酸酶降解能力、CCK-8法检测MCS-PLGA-NPs的细胞毒性、激光共聚焦观察巨噬细胞对MCS-PLGA-NPs-质粒DNA复合物的摄取、荧光显微镜和Western blot验证MCS-PLGA-NPs加载质粒DNA在细胞中的表达。元素分析结果表明，成功制备了取代度为5%~10%的MCS。纳米粒径测定和扫描电镜结果表明，MCS-PLGA-NPs的zeta为正值、粒径分布均匀且形态规则呈球形。琼脂糖凝胶电泳结果显示，MCS-PLGA-NPs吸附质粒的能力随着其质量的增加而增强并且可以在一定程度上抵抗核酸酶降解质粒DNA。在细胞毒性试验中，不同浓度的MCS-PLGA-NPs与RAW264.7细胞共孵育24 h后，细胞存活率仍在85%以上。在细胞摄取试验中，用激光共聚焦显微镜可以明显观察到质粒DNA结合到纳米微球表面被RAW264.7细胞摄取。荧光显微镜和Western blot试验证明MCS-PLGA-NPs加载质粒DNA可以在细胞中进行表达。综上表明，本研究成功制备了MCS以及具有递送核酸疫苗能力的MCS-PLGA-NPs，为FMDV核酸疫苗的递送研究提供了新的方向和见解，也为该递送载体携带特定抗原靶向抗原递呈细胞表面甘露糖受体以及应用于动物免疫的研究奠定基础。