长角血蜱酸性磷酸酶全长cDNA的克隆与原核表达及其表达产物的酶活性分析

通过对长角血蜱饥饿雌性成蜱cDNA表达文库中筛选到的未知基因进行5′RACE扩增,得到了长角血蜱酸性磷酸酶全长cDNA序列。利用生物信息学方法对该基因的核苷酸序列和推导的氨基酸序列进行了分析,同时构建了系统发育树。采用PCR方法扩增目的基因完整的开放阅读框,将其克隆到表达载体pGEX-4T-1中,构建重组质粒,转入BL21(DE3)中,经IPTG进行诱导表达,分别用SDS-PAGE和Wes-tern-blot分析目的蛋白的表达情况和反应原性,同时进行体外酶活性分析。结果表明,成功获得了长角血蜱酸性磷酸酶全长序列,体外高效诱导表达了约67.0 ku的重组蛋白。Western-blot表明该蛋白具有很强的反应原性,且在pH5.0时,该酶水解对硝基苯磷酸二钠的能力最强。     

WHEN PARENTS CAN CHOOSE TO HAVE THE “PERFECT” CHILD: WHY FERTILITY CLINICS SHOULD BE REQUIRED TO REPORT PREIMPLANTATION GENETIC DIAGNOSIS DATA

Preimplantation Genetic Diagnosis (PGD) is a procedure used to screen embryos for certain genetic conditions before implantation via in vitro fertilization (IVF) so the desired embryos can be chosen for implantation. The procedure was originally used to prevent the birth of children with deadly genetic disorders, but it is now used for more controversial reasons, such as to select for sex. Limited information is available regarding how PGD is used in the United States and there are many ethical concerns surrounding the practice of PGD, most notably that it could lead to the creation of designer babies and eugenic practices. This Note proposes amending a federal act to require fertility clinics to report PGD data through an existing web‐based system. This data can then be used by policymakers to federally regulate PGD practices.     

联合应用常染色体STR和X-STR标记鉴定单亲疑难案例

目的通过对常染色体和X染色体遗传标记的检测,探讨单亲疑难案例的鉴定策略。方法提取3个单亲鉴定案例的6份血样,采用Goldeneye 20A试剂盒和AGCU21＋1试剂盒检测常染色体上39个STR基因座,采用自主研制的16重X-STR扩增系统检测X染色体上16个STR基因座。结果用Goldeneye 20A试剂盒检测后发现每个单亲案例均有一个基因座不符合遗传规律,当常染色体STR基因座增加到39个时,案例1累计出现3个矛盾基因座;案例2和案例3均没有出现新的矛盾基因座。X染色体STR分型结果显示案例1有8个矛盾基因座,案例2和案例3无矛盾基因座,与常染色体分型结论相符。结论对于出现单基因座不符合遗传规律的母女、母子、父女单亲案例鉴定,不仅可以增加新的常染色体STR检测,也可以增加X染色体STR的检测,这样在相互验证的同时也能获得更加可靠的鉴定意见。     

LC-MS/MS鉴定大鼠血浆中桃红四物汤有效成分

目的 研究桃红四物汤有效成分体内作用的物质基础。方法 利用液相色谱-串联质谱（liquid chromatography technique coupled with tandem mass spectrometry,LC-MS/MS）提供的色谱及离子碎片信息,对大鼠灌胃桃红四物汤中入血成分进行分析鉴定。结果 通过保留时间比对、离子碎片解析等综合分析,鉴定桃红四物汤中6个入血成分（梓醇、藁本内酯、阿魏酸、芍药苷、芍药内酯苷和苦杏仁苷）均为原形成分,入血成分可能为桃红四物汤体内直接作用的有效成分。结论 本实验确定了桃红四物汤经口服后可被胃肠道吸收入血的6个有效成分,为后期桃红四物汤的药物代谢动力学研究提供了参考依据。     

茜草地上部分的化学成分研究

目的 研究茜草（Rubia cordifolia L.）地上部分化学成分。方法 利用硅胶柱色谱、反相硅胶柱色谱、Sephadex LH-20凝胶柱色谱等多种方法进行分离纯化;根据理化性质和谱学数据对分离得到的化合物进行结构鉴定。结果 分离得到10个单体化合物,分别鉴定为乙酰齐墩果酸（1）、2-羟基-1-甲氧基蒽醌（2）、β-谷甾醇（3）、（-）-表丁香脂素（4）、反式对羟基肉桂酸（5）、对苯二甲酸二丁酯（6）、（-）-3,4-二香草基四氢呋喃（7）、东莨菪亭（8）、异落叶松脂素（9）、5,5′-二甲氧基-7-氧代落叶松脂醇（10）。结论 化合物1～10为首次从茜草地上部分中分离得到,化合物1～2、4～10为首次从本植物中分离得到。     

Military genomic testing: proportionality,expected benefits,and the connection between genotypes and phenotypes

Mehlman and Li offer a framework for approaching the bioethical issues raised by the military use of genomics that is compellingly grounded in both the contemporary civilian and military ethics of medical research, arguing that military commanders must be bound by the two principles of paternalism and proportionality. I agree fully. But I argue here that this is a much higher bar than we may fully realize. Just as the principle of proportionality relies upon a thorough assessment of harms caused and military advantage gained, the use of genomic research, on Mehlman and Li''s view, will require an accurate understanding of the connection between genotypes and phenotypes – accurate enough to ameliorate the risk undertaken by our armed forces in being subject to such research. Recent conceptual work in evolutionary theory and the philosophy of biology, however, renders it doubtful that such knowledge is forthcoming. The complexity of the relationship between genotypic factors and realized traits (the so-called ‘G→P map’) makes the estimation of potential military advantage, as well as potential harm to our troops, incredibly challenging. Such fundamental conceptual challenges call into question our ability to ever satisfactorily satisfy the demands of a sufficiently rigorous ethical standard.     

柱前衍生化反相高效液相色谱法测定蝉蜕中3种氨基酸含量

目的 建立柱前衍生化反相高效液相色谱法测定蝉蜕中天冬氨酸、酪氨酸和缬氨酸3种氨基酸含量的方法。方法 采用酸水解法制备氨基酸样品溶液,以异硫氰酸苯酯柱前衍生化后进行高效液相色谱分析。采用Kromasil C18色谱柱(250 mm×4.6 mm),梯度洗脱,流动相A为0.1 mol/L醋酸钠缓冲液- 乙腈(93∶7),流动相B为乙腈- 水(4∶1),流速1.0 mL/min,柱温为43 ℃,检测波长254 nm,进样量20 μL。结果 氨基酸衍生溶液在36 h内保持稳定,3种氨基酸在线性范围内峰面积与浓度的相关系数均大于0.999 0,加样回收率(n＝6)分别为95.62%、101.26%和100.00%。结论 所建立的检测方法稳定可靠,精密度和重复性良好,可用于蝉蜕中天冬氨酸、酪氨酸、缬氨酸含量测定。     

Lipidic compounds found in soils surrounding human decomposing bodies and its use in forensic investigations – A narrative review

Following decomposition of a human body, a variety of decomposition products, such as lipids, are released into the surrounding environment, e.g. soils. The long-lasting preservation in soils and their high diagnostic potential have been neglected in forensic research. Furthermore, little is known about the preservation, chemical transformation, or degradation of those human derived lipids in soils. To date, several studies identified various lipids such as long-chain free fatty acids and steroids in soils that contained decomposition fluids. Those lipids are preserved in soils over time and could serve as markers of human decomposition in forensic investigations, e.g. for estimating the post-mortem interval or identifying the burial location of a human body. Therefore, this review focuses on the current literature regarding fatty acid and steroid that have been detected in soils and associated with human body decomposition. After a short introduction about human decomposition processes, this review summarises fatty acid and steroid analysis applied in current case studies and studies related to taphonomic research. This review provides an overview of the available studies that have used fatty acids and steroids as identifiers of human decomposition fluid in soils in a forensic context and discusses the potential for developing this innovative field of research with direct application in a forensic context.     

小儿智力咀嚼片提取工艺研究

目的优选小儿智力咀嚼片的最佳提取工艺。方法以远志酸的含量为指标,利用正交设计法考察溶剂种类、溶剂用量、提取时间以及提取次数对实验的影响,确定最佳提取工艺参数。结果最佳提取工艺参数为:以60%乙醇为溶剂,加10倍量,提取0.5 h,提取2次。结论该工艺可用于小儿智力咀嚼片的提取。