柔嫩艾美耳球虫DF-1细胞培养体系的建立及应用

为了建立柔嫩艾美耳球虫的鸡胚成纤维传代细胞(DF-1)培养体系,并应用于柔嫩艾美耳球虫子孢子的细胞入侵活力检测,将CFDA-SE标记的子孢子接入DF-1细胞,利用流式细胞仪测定子孢子的细胞入侵活力。比较了37℃和41℃培养温度下,子孢子对DF-1、MDBK、Vero、BHK、MDCK五种细胞的入侵活力。优化了在子孢子入侵活力检测时,DF-1细胞最佳培养条件。结果表明,柔嫩艾美耳球虫子孢子在DF-1细胞中可发育为第一代裂殖子。在不同培养温度下,子孢子对DF-1细胞的入侵活力均高于其他细胞。优化后的DF-1细胞培养体系的培养程序为:用含100mL/L胎牛血清的DMEM稀释DF-1细胞,以每孔1×105个细胞铺被24孔板,37℃、50mL/L CO2培养24h后,用含50mL/L胎牛血清的DMEM换液后,每孔接入3×105个子孢子,41℃、50mL/L CO2培养8～12h,收集细胞进行检测。     

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.     

三个柔嫩艾美球虫虫株线粒体cox 1基因部分序列的比较分析

为研究柔嫩艾美球虫不同毒力株的线粒体细胞色素c氧化酶第1亚基(cox1)基因与球虫种群之间的遗传关系,应用聚合酶链反应扩增柔嫩艾美球虫3个不同毒力株的cox1序列,并与GenBank上登录的鸡柔嫩艾美球虫、巨型艾美球虫、毒害艾美球虫和堆形艾美球虫虫株的相应序列进行比对分析.结果显示,每个虫株都获得659 bp的cox1部分有效序列(pcox1).柔嫩艾美球虫不同虫株的pcox1序列完全相同,但与其他种的艾美球虫相应的pcox1序列有不同程度的差异.表明柔嫩艾美球虫的cox1序列可作为艾美球虫不同种虫株之间遗传变异研究的标记.     

Determinants and Evaluation of Local Energy‐Efficiency Initiatives from the American Recovery and Reinvestment Act

This article examines variation in local‐level energy‐efficiency grants and corresponding initiatives from the American Reinvestment and Recovery Act (ARRA) in the United States. The analysis is based upon a hurdle model of counts of energy‐efficiency grants received by 348 local governments that received these grants from 2009 to 2013, as well as 348 matched local governments that did not receive such funds. City‐level characteristics including amount of federal financial support, per capita income, signaling of preferences for sustainability policies, manufacturing, and political influences are shown to be empirically important determinants of variation in local energy‐efficiency initiatives. The evidence suggests that all else held equal, the $21.8 billion in ARRA funds expended with the intent of increasing local energy‐efficiency programs and policies successfully led to this end.     

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.     

消瘀接骨散对兔膝骨关节炎模型关节液中MMP-1、MMP-3表达的影响

目的观察消瘀接骨散对兔膝骨关节炎模型中关节软骨病理改变及关节液中基质金属蛋白酶(matrix metalloproteinase,MMP)-1、MMP-3表达的影响。方法将24只新西兰大耳白兔分为正常组、模型组、用药组,每组8只。采用膝关节石膏制动方法复制膝骨关节炎模型,模型复制成功后,正常饲养并驱赶1周,给予用药组家兔消瘀接骨散外敷,给予模型组和正常组家兔相同大小药棉、绷带、胶布固定。光镜下观察膝关节软骨退变情况,采用酶联免疫吸附法测定关节液中MMP-1、MMP-3表达水平。结果用药组家兔膝关节软骨损伤较模型组明显减轻,关节软骨表面较正常组粗糙,滑膜增厚、稍有肿胀,镜下可见细胞排列规律尚可,偶见少量细胞簇集现象,关节软骨细胞有轻度变性,潮线尚完整。用药组家兔膝关节软骨的Mankin评分及关节液中MMP-1、MMP-3表达水平明显低于模型组(P0.05)。结论消瘀接骨散外敷治疗兔膝骨关节炎的机制与其降低关节液中MMP-1、MMP-3的表达水平有关。