杀虫双的检验和质谱解析

本文用气相色谱/质谱联用方法检验杀虫双,并解析杀虫双的质谱图.     

猪圆环病毒2型感染性克隆的构建

为进一步研究猪圆环病毒2型(PCV2)的致病机制和基因功能,利用PCR方法从河北保定株(HBBD0608)猪PCV2病料中获得了PCV2全基因组序列,将其定向克隆入pEGFP-N1载体,成功构建了PCV2全基因组串联双拷贝的重组质粒pEGFP-N1-2PCV2.将该质粒转染PK-15细胞并连续盲传5代,用PCR方法可以在转染后盲传的细胞中检测到PCV2核酸;间接免疫荧光可以检测到绿色荧光,表明有PCV2抗原存在.证明构建的双拷贝重组质粒具有感染性.     

猪PD-1分子及其配体的实时荧光定量RT-PCR检测方法的建立

根据猪程序性死亡因子PD-I及其配体PD-LI和PD-L2的基因序列,分别设计了特异性引物和TaqMan探针,以10倍系列稀释的重组质粒pMD-PD-1、pMD-PD-L1和pMD-PD-L2为标准品,对实时荧光定量PCR的条件进行优化,以建立定量检测这3个基因的方法.结果表明,建立的方法在1×10~2～1×10~8copies/μL模板范围内具有良好的线性关系(r~2>0.99),扩增效率均高于96%,可检测至少100 copies的阳性标准品.利用该方法对猪外周血单核细胞中这3个基因的表达情况进行了检测,结果也表明该方法具有良好的敏感性、特异性和重复性,可应用于临床样品的检测.     

猪戊型肝炎病毒广西株ORF2三段连续基因编码蛋白的表达及抗原性分析

为了确定猪戊型肝炎病毒(HEV)ORF2抗原性片段的分布,参考中国T1株序列设计套式引物,从新鲜猪粪中扩增出HEV部分基因片段,长1725bp,位于ORF2第205～1929bp之间,命名为HEVORF2-A。根据HEVORF2-A序列设计了3对引物,分别扩增HEVORF2-A三段连续的基因片段,命名为HEVORF2-B1、HEVORF2-B2、HEVORF2-B3,分别位于HEVORF2-A第1～576bp,577～1149bp,1150～1725bp之间。将这4段扩增的基因片段分别与表达载体pET32a(+)连接,并转入BL21(DE3)pLysS感受态细胞,经IPTG诱导,SDS-PAGE分析。结果显示,经1mmol/LIPTG37℃诱导6h后,除pET32a-A在82.6ku处有微量融合蛋白表达外,pET32a-B1、pET32a-B2、pET32a-B3分别在41.3ku、41.5ku、41.8ku处有高水平的融合蛋白表达。用自然感染猪血清和北京万泰HEVELISA诊断试剂盒阳性对照血清进行Western-blot分析,结果3个短的融合蛋白均能检测到明显的条带,表明构建的3个表达载体具有良好的抗原性。     

冷应激对雏鸡空肠黏膜前列腺素E2含量和环氧合酶-2 mRNA表达的影响

以15日龄健康雏鸡为试验对象,进行急性和慢性冷应激(12±1)℃处理,检测空肠黏膜前列腺素E2(PGE2)含量和环氧合酶-2(COX-2)mRNA的表达量。结果显示,急性冷应激时空肠黏膜PGE2含量和COX-2mRNA的表达量无显著变化;慢性冷应激时,空肠黏膜PGE2含量和COX-2mRNA表达量均显著升高,并随着冷应激时间的延长而降低。说明冷应激可引起雏鸡空肠黏膜PGE2含量和COX-2mRNA表达量的变化,这些变化可能在冷应激对肠道的损伤中起一定的作用。     

从“北溪-2”项目看德俄关系及俄美欧三方博弈

作为两个重要的地区大国,德国与俄罗斯的关系对彼此和欧洲都有着重要影响。在双边关系中,能源因素因其对国民经济生产的决定性意义长期发挥着“稳定器”的作用。2018年9月,由德俄牵头,法国、奥地利与荷兰合资建造的“北溪-2”天然气管道项目正式动工铺设。该项目将使输往欧洲的俄罗斯天然气绕开乌克兰,经过波罗的海海底直接连通德俄两国,德国由此成为俄罗斯天然气最重要的转运国。此举虽然在双边层面上保证了德俄的能源经济安全,但在多边层面却招致欧盟、美国等多方反对。因担心会进一步加深对俄罗斯能源的依赖,欧盟委员会及中东欧国家欲诉诸法律对该项目施以约束;美国在制裁项目企业的同时转而向波兰、乌克兰等国出口液化天然气,在利用政治机遇抢占欧洲市场的同时试图制衡俄罗斯的影响力。在多方力量博弈中,德国需要在平衡俄美欧关系中付出更多努力。     

Prodrugs of New Psychoactive Substances (NPS): A New Challenge

The concept of a substance acting as a prodrug for an intended drug is not new and has been known and utilized with particular benefits within medicine for efficacy and patient safety. Prodrugs of psychoactive substances are also not particularly new but this has also extended to considerations of prodrugs of new psychoactive substances (NPS). The continuing evolution of NPS has been a constant forensic challenge. In some countries, this constant evolution has led to the introduction of various alternative methods of drug control. Whether for this reason or in the pursuit of user experimentation, prodrugs of NPS have been discussed, developed, and exploited, posing some distinct forensic challenges. This is especially the case within toxicological analysis of biological fluids and for some substances, also forensic chemical analysis, through inherent instability of the prodrug or metabolism in the body. Particular examples of NPS prodrugs include 1-propanoyl-LSD, 1-butanoyl-LSD, 1-acetyl-LSD, and 2C-B-AN. This is in addition to associated substances and medicines that may be used for an intended pharmacological effect. Various prodrugs for stimulant and hallucinogenic substances in particular have appeared in the literature and have been discussed within drug user forums and made available for purchase online. Presently, drug monitoring data from national and international systems indicate that prodrugs are not widely available or problematic. Nevertheless, it is important that there is sufficient awareness of the prodrug concept and potential impact and associated forensic implications, not just for chemical analysis but also for toxicological considerations when a substance has been used.     

Integrating Concerns with Climate Change into Local Development Planning in Cambodia

Integration of climate change adaptation with development planning at multiple scales is widely seen as preferable to reactive, fragmented, or highly centralized responses. At the same time, there are growing concerns on when intervention is most appropriate, the transaction costs of coordination, and the adequacy of institutional capacity at local levels, especially in developing countries. This article examines entry points and mechanisms for integrating concerns with climate change into local development planning in Cambodia. An institutional ethnography of the planning process indicates that subnational planning is participatory and flexible; and thus, provides plausible entry points to integrate climate change concerns. Case study methods applied to two externally supported, climate‐resilient development projects identify promising mechanisms and strategies, as well as obstacles to integration. A vulnerability reduction assessment tool and top‐up grant scheme both included promising deliberative and participatory elements from which lessons for future and elsewhere can be drawn. At the same time, key stakeholders concede that local integration more widely is hampered by multiple obstacles, including weak institutional capacity, low community participation, and lack of resources and incentives. Addressing these challenges requires political commitments for good governance, capacity development, and additional resources.     

我国P2P平台准入制度的检讨与重构

目前,我国P2P平台准入制度尚存在准入方式不当、准入条件混乱和准入标准模糊等不足,导致P2P市场出现野蛮增长、良莠不齐、鱼目混珠的乱象。重构P2P平台准入制度,应遵循"理念--模式--制度"的路径,以"适度"为理念设置控制与激励双重标准,选择差异化准入监管模式,具化准入方式、条件和标准等实质性要件。