Sample Mining and Data Mining: Combined Real-Time and Retrospective Approaches for the Identification of Emerging Novel Psychoactive Substances

Novel psychoactive substances (NPS) are synthetic drugs that pose serious public health and safety concerns. A multitude of NPS have been identified in the United States, often implicated in forensic investigations. The most common and effective manner for identifying NPS is by use of mass spectrometry and the true utility lies within nontargeted acquisition techniques. During this study, a liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) assay was developed, validated, and implemented for forensic toxicology testing. A SCIEX TripleTOF™ 5600 + with SWATH^® acquisition was used. Resulting data were compared against an extensive library database containing more than 800 compounds. The LC-QTOF-MS assay was applied to the reanalysis of biological sample extracts to discover emergent NPS. More than 3,000 sample extracts were analyzed, and more than 20 emerging NPS were detected for the first time. Among these were isopropyl-U-47700, 3,4-methylenedioxy-U-47700, fluorofuranylfentanyl, N-methyl norfentanyl, 2F-deschloroketamine, 3,4-methylenedioxy-alpha-PHP, eutylone, and N-ethyl hexedrone.     

Applying Stone in a Western Landscape: Ranchers,Conservationists, and Causal Stories in the “American Serengeti”

As the emergence of nongovernmental conservation efforts generates conflict among various stakeholders, the causal story that each party articulates regarding conservation and the causes of land degradation reflects their unique interests. This study uses existing literature to evaluate causal stories surrounding a contemporary conservation effort: Montana's American Prairie Reserve. Through qualitative review of web‐based documents and newspaper articles, it generates a preliminary account of key stakeholders' causal stories. The case study suggests that parties who might be disadvantaged by ascribing responsibility for environmental harms in an adversarial fashion may instead elect to articulate causal stories that are more neutral than existing approaches might forecast. The study concludes by suggesting that further development of causal story literature may enable it to better address contemporary conservation efforts.     

中国汉族与日本群体DYFl55S1基因座的遗传多态性

目的探讨Y染色体DYF155S1基因座的遗传多态性及群体间差异。方法 应用MVR-PCR、荧光显谱及DNA序列分析技术,对来自中国群体(北方汉族,64例)和日本群体(43例)男性个体的DYF155S1基因座进行初步分析。结果107例样本共检出了5种重复序列类型,包括新的命名为6型的重复序列,它是在1型的基础上T22A置换所形成,仅存在于日本群体,可作为民族特征性遗传标记。2群体重复序列的排列方式以3134顺序为主,在中国和日本群体中各占73.44％和67.44％,是黄种人的特点。134顺序在中国群体中占第二位,为17.19％,6134排列占日本群体的16.28％。3’端的4型重复序列的平均数目在日本群体为8.8条,明显低于中国群体的12.5条。结论DYF155S1基因座具有非常高的遗传多态性和明显的群体差异。     

大鼠液压冲击脑损伤TGF-β1及其受体TβR I的表达

目的观察脑损伤后TGF-β1及其受体TβRI蛋白表达及其时序性变化,探讨脑损伤的分子机制及法医学脑损伤时间推断。方法用免疫组织化学方法观察大鼠液压冲击脑损伤后TGF-β1及其受体TβRI蛋白在伤后不同时间(30 min、1 h、3 h、6 h、12 h、1 d、3 d、7 d)的表达情况,以正常大鼠及手术大鼠作为对照。结果 (1)正常及手术对照组大鼠脑组织内有低水平的TGF-β1及TβRI表达;(2)脑损伤后1～3d,大脑皮质和脑干TGF-β1/TβRI蛋白表达逐渐增加,7 d时仍维持在高表达水平;(3)海马冲击后12 h～1 d逐渐增加,3 d时仍处于高表达水平,7 d时已开始下降。结论脑损伤可诱导TGF-β1/TβRI基因在脑内表达,其时序性改变可望用于法医学脑损伤时间推断。     

鸡白细胞介素-2受体基因γ链的转录分析与表达

以脾单个核细胞总RNA为模板,对鸡白细胞介素-2受体基因γ链(chIL-2Rγ)进行了RT-PCR,获得了一1 047 bp的开放阅读框,编码由348个氨基酸残基组成的分子质量为37.8 ku的蛋白多肽。预测的鸡IL-2Rγ多肽链中包含4个保守半胱氨酸残基、1个WSXWS基序和7个N连接的糖基化位点。鸡IL-2Rγ与其他动物IL-2Rγ在氨基酸水平上的同源性仅为21.4%~38.2%。RT-PCR检测发现,鸡IL-2RγmRNA分布于大脑、小脑、脊髓、腔上囊、脾、胸腺、骨髓、盲肠扁桃体、腺胃、肌胃、空肠、卵黄囊憩室、回肠、盲肠、直肠、心脏、肾、肺、肝、骨骼肌和皮肤,而在十二指肠和睾丸中没有检测到其转录。构建了鸡IL-2Rγ胞外区的原核重组表达载体,进行了表达和鉴定。     

堆型艾美球虫广东株3-1E基因的克隆与序列分析

根据GenBank中登录的堆型艾美球虫31E基因序列,设计了3条引物,以广东株堆型艾美球虫裂殖子总RNA为模板,利用反转录聚合酶链反应(RTPCR)扩增获得了31E基因部分片段,将这一片段克隆至pGEMTEasy载体中,经PCR、限制性内切酶分析和克隆片段的序列测定、比较,证实了克隆片段的可靠性。序列比较发现,所克隆的基因片段与Eimeriaacervulina美国株(US)、E.acervulinaQH株31EcDNA的核苷酸同源性分别为99.0%和99.2%,推导氨基酸的同源性分别为98.2%和97.6%。     

鸡肾型传染性支气管炎病毒的分离鉴定

从陕西省榆林市某蛋用种鸡场疑似肾型传染性支气管炎病鸡中分离到了1 株肾型IBV(定名为YL 04),并对分离病毒进行了血凝性、血凝抑制性、致病性、鸡胚矮小化、电镜特征等生物学特性鉴定及S1基因5′端的RT PCR鉴定。结果表明,该分离株经10 g/L胰酶处理后的各代病毒尿囊收集液均可凝集鸡红细胞;标准阳性血清可特异性地抑制其凝集性;可复制出与自然发病相同的病例;病毒传代物有明显的致鸡胚矮小化作用;透射电镜下可见有近似球形、直径100 nm左右的冠状病毒粒子;用RT PCR方法扩增到1 条373 bp的目的片段,其核苷酸序列与IBV CQ/01/2004株序列的同源性达98%。     

鸭传染性腔上囊病毒VP2基因的克隆与序列分析

应用RT PCR法对分离于当地典型发病鸭群的鸭传染性腔上囊病毒 (IBDV )YL997株进行了VP2基因的克隆与序列分析 ,并和相关毒株进行了比较。结果表明 ,YL997株与STC株的核苷酸和氨基酸同源性只有 92 .5 %和 91.8% ,与超强毒株UK6 6 1的核苷酸和氨基酸同源性均为97.6 % ,而与当地鸡群中分离的超强毒株HN94 2的核苷酸和氨基酸同源性则高达 98.1%和98.4 % ,该毒株的VP2基因序列完全具备了超强毒株的主要特征。     

猪圆环病毒2型感染裸鼠及BALB/c小鼠的研究

用猪圆环病毒2型(PCV2)经腹腔注射BALB/c小鼠及裸鼠各15只,每隔2周1次,共感染3次。BALB/c小鼠初次、再次感染后均未出现明显的临床症状,仅有2只小鼠出现病理变化;从血清中可检测到PCV2核酸及其ORF2抗体;同时淋巴细胞增殖反应显著增强,NK和CTL细胞杀伤率显著升高;CD4+、CD8+、CD3+T淋巴细胞及CD19+B淋巴细胞数量显著减少。裸鼠也未出现明显的临床症状,从血清中可检测到PCV2核酸,但未检测到PCV2 ORF2抗体;除NK细胞杀伤率显著升高外,其余免疫学指标无显著改变。结果表明,BALB/c小鼠比裸鼠对PCV2易感,各项免疫学指标变化与PCV2感染猪一致,但不表现临床症状,仅个别BALB/c小鼠出现病理变化,说明BALB/c小鼠对PCV2不如猪易感。