Identification of a conservative site in the African swine fever virus p54 protein and its preliminary application in a serological assay

BackgroundASF was first reported in Kenya in 1910 in 1921. In China, ASF spread to 31 provinces including Henan and Jiangsu within six months after it was first reported on August 3, 2018. The epidemic almost affected the whole China, causing direct economic losses of tens of billions of yuan. Cause great loss to our pig industry. As ELISA is cheap and easy to operate, OIE regards it as the preferred serological method for ASF detection. P54 protein has good antigenicity and is an ideal antigen for detection.ObjectiveTo identify a conservative site in the African swine fever virus (ASFV) p54 protein and perform a Cloth-enzyme-linked immunosorbent assay (ELISA) for detecting the ASFV antibody in order to reduce risks posed by using the live virus in diagnostic assays.MethodWe used bioinformatics methods to predict the antigen epitope of the ASFV p54 protein in combination with the antigenic index and artificially synthesized the predicted antigen epitope peptides. Using ASFV-positive serum and specific monoclonal antibodies (mAbs), we performed indirect ELISA and blocking ELISA to verify the immunological properties of the predicted epitope polypeptide.ResultsThe results of our prediction revealed that the possible antigen epitope regions were A23–29, A36–45, A72–94, A114–120, A124–130, and A137–150. The indirect ELISA showed that the peptides A23–29, A36–45, A72–94, A114–120, and A137–150 have good antigenicity. Moreover, the A36–45 polypeptide can react specifically with the mAb secreted by hybridoma cells, and its binding site contains a minimum number of essential amino acids in the sequence 37DIQFINPY44.ConclusionsOur study confirmed a conservative antigenic site in the ASFV p54 protein and its amino acid sequence. A competitive ELISA method for detecting ASFV antibodies was established based on recombinant p54 and matching mAb. Moreover, testing the protein sequence alignment verified that the method can theoretically detect antibodies produced by pigs affected by nearly all ASFVs worldwide.     

高温胁迫下绿豆象内参基因筛选及其Hsp70基因的表达特性

为明确Hsp70蛋白在绿豆象Callosobruchus chinensis高温耐受性中的作用,该研究筛选并克隆β-actin、α-tubulin、β-tubulin、EF1-α、GAPDH、Hsc70、AK和RPL40八个候选内参基因,采用实时荧光定量PCR(quantitative real-time PCR,qPCR)技术测定其在绿豆象雌雄成虫体内的表达量,通过geNorm、BestKeeperr和NormFinde软件对这8个候选内参基因的表达稳定性进行评价,选择最适宜的内参基因对不同高温胁迫下绿豆象体内Hsp70基因的表达特性进行测定。结果表明,8个候选基因引物均有良好的特异性和引物扩增效率,引物扩增效率介于91.6%～108.1%之间。高温胁迫下,β-tubulin基因较其他7个候选基因有较小的平均变异度、稳定值和标准差,为最适宜的内参基因。39、42和45℃高温处理1 h后绿豆象雌雄成虫体内Hsp70表达水平较对照显著上调,当处理时间延长至3 h后,Hsp70基因的相对表达量较对照也有不同程度上调,但差异不显著,表明绿豆象Hsp70基因可以响应高温胁迫,Hsp70蛋白在绿...     

中国条锈菌新小种条中30、31号的研究

本文报道了1991年以来对新小种条中30、31号鉴定与致病性的研究。继1991年发现对绵阳系成株小麦有致病力的、对Hybrid46有毒的新致病类型91—1,1993年又发现了新的致病类型93—1。根据它们对我国鉴别寄主的反应,命名为条中30、31号。与条中28、29号相比,新小种具有更宽毒性基因组成和更高的相对寄生适合度值,它们对我国生产品种、高代品系和重要抗源有更广的致病范围。证实两个新小种的出现和发展是绵阳系小麦抗条锈变异的主要因素,建议加强对新小种抗病育种和流行预测的研究。     

糖苷水解酶家族基因AghyGH1a参与莲草直胸跳甲对皂苷的代谢

为了解植物化合防御物质皂苷对专食性天敌莲草直胸跳甲Agasicles hygrophila的防御作用,测定莲草直胸跳甲2龄幼虫取食皂苷后的排泄和肠道变化,利用转录组测序筛选参与皂苷代谢的候选基因,并采用 RNA 干扰技术鉴定候选基因功能。结果发现,取食皂苷导致莲草直胸跳甲幼虫排泄量增加,肠道畸变,排泄量与取食皂苷浓度正相关。转录组测序发现,与对照相比,取食 100 mmol/L皂苷处理的莲草直胸跳甲幼虫肠道有496条差异表达基因,其中261个上调基因和235个下调基因。GO分类和KEGG分类分析发现糖苷水解酶家族1（glycoside hydrolase family1,GH1）基因集中在碳水化合物水解通路。利用RNA干扰技术鉴定发现,莲草直胸跳甲AghyGH1a基因沉默可导致幼虫取食皂苷后排泄量增加,肠道内容物颜色更暗沉。表明莲草直胸跳甲AghyGH1a参与皂苷代谢。     

猪轮状病毒感染研究进展

猪轮状病毒感染是由猪轮状病毒引起的一种传染病,多发生于仔猪。其典型症状为腹泻。此病分布广泛,引起严重经济损失。本文对病原学、流行病学、临床症状、发病机理、免疫、诊断、防制等方面的研究进展进行了综述。     

Oncolytic Vaccinia Virus Carrying Aphrocallistes vastus Lectin (oncoVV-AVL) Enhances Inflammatory Response in Hepatocellular Carcinoma Cells

Aphrocallistes vastus lectin (AVL) is a C-type marine lectin derived from sponges. Our previous study demonstrated that oncolytic vaccinia virus carrying AVL (oncoVV-AVL) significantly enhanced the cytotoxicity of oncoVV in cervical cancer, colorectal cancer and hepatocellular carcinoma through the activation of Ras/ERK, MAPK/ERK and PI3K/Akt signaling pathways. In this study, the inflammatory response induced by oncoVV-AVL in a hepatocellular carcinoma cell (HCC) model was investigated. The results showed that oncoVV-AVL increased the levels of inflammatory cytokines including IL-6, IL-8 and TNF-α through activating the AP-1 signaling pathway in HCC. This study provides novel insights into the utilization of lectin AVL in the field of cancer therapy.     

次生湿地水生植物应用评价

种植水生植物是优化水生生物群落,提高水生态系统的自净能力治理及修复湿地水体的有效途径。该文针对南部次生沉降湿地水环境的质量特征,采用秩和比与特尔菲法相结合的方法,对在唐山地区应用广泛的21种水生植物进行综合评价。研究表明:综合效能高的水生植物应确立为南湖次生沉降湿地绿化建设的首选植物材料,综合效能较低的水生植物,再利用时要充分考虑其特性,谨慎使用。     

笼型异步磁力耦合器机械特性与试验

为有效解决目前机械传动系统中选用电机时功率裕度过大而普遍存在的能源利用效率较低问题,提出一种基于三相异步电机鼠笼转子的异步磁力耦合器(squirrel cage asynchronous magnetic coupler,SCAMC)。结合SCAMC具体结构特点,采用标量磁位法及二维场边界条件,建立气隙磁场数理模型;在气隙磁通密度中引入时间变量,推导出感生电流随时间变化的表达式;基于电流叠加性,将笼条电流折算到转子表面,并沿圆周方向对感生电流所形成的洛伦兹力进行积分,建立了SCAMC的电磁转矩模型。基于上述理论及技术基础,设计并制造出一台37 k W SCAMC样机,并对其机械特性进行理论计算、仿真验证及试验测试。结果表明:转差率相同时,所得的仿真及试验数据与理论计算值相比,误差不超过5%;SCAMC与同容量的三相异步电机相比,线性工作区更宽,过载能力更强,但其机械特性偏软,可有效缓解负载对电机的冲击。该研究可为磁力耦合器在大惯量、难启动及经常性过载机械设备中的应用提供参考。     

新疆大蒜叶疫病的研究:Ⅱ病菌越冬致病性病害流行因素及防治

 试验结果表明,本菌(Stemphylium vesicarium(Wallr.) Simons)以病残体上分生孢子和菌丝体在田间和冷藏室越冬,9月至翌年6月检查,分生孢子萌发率为16.4~41.3%。人工接种试验指出,不论有无伤口,病菌均可侵染大蒜,但洋葱与大葱在伤口接种条件下病情较重。在自然条件下,大蒜的斑点型病状出现于6月下旬,7月为病情高峰期。影响病情的天气因素,主要是7月份温度和7月中旬的降水量(r=0.69)。如果7月雨次多、雨时长,病情即极为严重,可导致蒜田绝收。化学防治试验结果,以腐霉利防效最佳。从7月上旬开始,每间隔10~13天喷药1次。喷药2~3次即可取得较好的防治效果。     

壬菌铜30%微乳剂防治葡萄霜霉病田间药效试验

试验表明壬菌铜30%微乳剂对葡萄霜霉病有很好的防治效果,使用浓度为200、300mg/kg时,对葡萄霜霉病的防治效果分别为79.9%、85.3%,与氢氧化铜46%水分散粒剂230mg/kg、烯酰吗啉40%悬浮剂250mg/kg处理的防效相当,且耐雨水冲刷,持效期长,价格低廉,值得推广使用。