基因兴奋剂检测的现状与展望

生物技术在取得巨大成就的同时也导致了兴奋剂的发展。从体外重组蛋白到潜在的基因兴奋剂的出现,给兴奋剂检测工作带来了巨大的挑战。基因治疗的发展以及一些功能基因对运动技能提高的潜力,加大了基因兴奋剂使用的可能性。本文在介绍基因兴奋剂的基础上,阐述了基因兴奋剂检测方法的研究现状,展望了基因兴奋剂检测技术的发展策略。     

基因兴奋剂检测方法研究进展

基因兴奋剂是指以非治疗目的而向运动员体内导入外源基因或细胞等物质,以不正当方法提高运动员成绩的物质或技术。目前随着基因治疗技术的发展,基因兴奋剂被用于竞技体育赛场的潜在问题受到越来越密切的关注,因此对基因兴奋剂检测方法的需求也愈发迫切。本文简要介绍了基因兴奋剂的潜在候选基因、导入途径与载体、基因兴奋剂的危害以及已有基因兴奋剂的检测方法,并对未来基因兴奋剂检测领域急需解决的关键问题进行了讨论。     

高效毛细管电泳在兴奋剂检测中的应用进展

对高效毛细管电泳(HPCE)在兴奋剂检测方面的工作进行了评述,讨论了高效毛细管电泳不同分离模式在兴奋剂检测中的应用,并对这一领域的发展趋势进行了展望。     

赛默飞世尔科技(原热电公司)与中国兴奋剂检测中心签署2008年奥运会兴奋剂检测项目合作计划书

赛默飞世尔科技(原热电公司)与中国兴奋剂检测中心在北京签署2008年奥运会兴奋剂检测项目合作计划书。协议确定了赛默飞世尔参与和支持2008年奥运会兴奋剂检测的仪器和技术服务,以保证奥运会期间兴奋剂的检测任务的完成。反兴奋剂是目前各国政府和体育界共同面临的问题,而赛默飞世尔科技与国际兴奋剂检测机构的合作可以追溯到世界反兴奋剂斗争的最开始。中国兴奋剂检测中心承担了2008年奥运会兴奋剂检测主要任务,先后购买了赛默飞世尔科技的DFS GC/MS高分辨磁式气相色谱质谱联用仪等多台高端质谱系统。同时,为实现2008北京奥运会期间兴奋…     

兴奋剂与兴奋剂检测研究进展

滥用兴奋剂会遭受国际奥委会的严厉处罚 ,但仍有不少运动员为冠军及奖牌所带来的巨大利益所诱惑 ,铤而走险 ,不惜以各种方式滥用兴奋剂 ,所以兴奋剂及其检测是体育界乃至全社会关注的问题。本文主要就兴奋剂、兴奋剂检测的概念以及该领域国内外的最新研究动态和发展方向进行介绍 ,并着重探讨肽类激素rhEPO、rhGH的研究方法和对不同检测基质 (尿样、发样、血样、体液 )的检测进展     

“兴奋剂检测方法专栏”序言

第29届奥运会即将于2008年8月8日在北京召开,伴随着中国社会的进步和科学、经济的腾飞,百年奥运梦想就要成真,华夏儿女无不为之欢欣。作为分析化学工作者,尤其是色谱工作者,我们在关注奥运本身的体育意义外,还会思考奥运精神以及与此相关的技术问题,其中兴奋剂检测就是一个不得不说的话题。 回顾现代奥林匹克运动的发展史,可以说始终伴随着滥用兴奋剂和反兴奋剂的斗争。随着体育运动的日益商业化,滥用兴奋剂的案例也越来越多。从1967年国际奥委会提出最早的禁用药物名单,到1968年初国际奥委会医学委员会正式宣布了特为法国格勒诺布尔冬奥会兴奋剂检查制定的禁用药物名单。从1999年国际奥委会瑞士洛桑成立世界反兴奋剂机构（WADA）,到在联合国教科文组织第33届会议通过《反对在体育运动中使用兴奋剂国际公约》(简称“反兴奋剂国际公约”),再到2007年第三届世界反兴奋剂大会通过修订后的《世界反兴奋剂条例》（将于2009年1月正式生效）,兴奋剂的种类越来越多,兴奋剂的检测技术越来越先进,真所谓“道高一尺,魔高一丈”。在某种程度上甚至可以说,分析化学的发展大大促进了反兴奋剂事业的进步,而反兴奋剂的需要,又推动了分析检测技术的更快发展。但无论如何,兴奋剂的滥用不仅严重背离了奥林匹克精神,而且会极大地损害相关运动员和教练员的身心健康。因此,为了配合北京奥运会的召开,为了让读者了解国际上兴奋剂检测研究的现状和我国科学工作者在兴奋剂检测领域的研究成果,也为了北京能办一届“干净”的奥运会,我们特别增设了“兴奋剂检测方法”专栏,共约稿13篇,其中有综述也有研究报告;有传统的检测方法也有最新的毛细管电泳和芯片技术检测兴奋剂的进展;在检测对象方面,既有小分子药物,也有大分子蛋白质;既有传统的药物,也有新的基因类兴奋剂,从不同的侧面反映了当今兴奋剂检测所面临的挑战和新型检测方法与技术大发展的展望。相信读者会从中获得有价值的信息。 非常感谢这么多作者欣然接受我们的邀请,把他们的最新研究结果呈现给我们。特别要感谢北京大学化学学院的白玉博士在稿件审阅和修改过程中所付出的大量劳动。我们愿意将这期专栏作为礼物献给2008年北京奥运会。     

β2-兴奋剂的样品前处理与分析检测方法研究进展

许多国家禁止β2-兴奋剂用于动物饲养,但违禁药品的使用屡有发生,因此β2-兴奋剂的分析检测引起了高度关注.该文综述了β2-兴奋剂分析的样品前处理方法,包括溶剂萃取、固相萃取、固相微萃取等,并对常用的β2-兴奋剂检测方法(如色谱分析法、电化学检测法、酶联免疫法、化学发光法等)的研究进展进行了评述.     

促红细胞生成素和人生长激素兴奋剂检测方法的研究进展

在2008年世界反兴奋剂组织颁布的兴奋剂目录中,S2项肽类激素及相关品种均属于内源性生物大分子物质,如何区分所检测物质属于外界摄入还是机体分泌是此项检测的重点与难点。本文针对其中应用最为普遍、研究较为深入的促红细胞生成素(EPO)和人生长激素(hGH)的检测,从间接血液指标检测、直接检测途径等方面进行了分类评述,侧重于从理化分析方法、免疫分析方法角度阐述识别及区分重组蛋白与内源性蛋白的新途径。     

兴奋剂重组人促红细胞生成素及其类似物检测方法的研究进展

重组人促红细胞生成素(rhEPO)是一种激素类兴奋剂,近年来被滥用在一些耐力性比赛项目中。由于重组与内源性EPO的氨基酸序列相同,区别很小,并且在尿样或血样中的浓度低,代谢快,给检测带来了很大的难度。本文从直接方法和间接方法两个方面综述了近几年来兴奋剂rhEPO及其类似物检测的研究进展,并结合本小组的工作展望了rhEPO检测的发展方向。     

液相色谱-质谱联用在兴奋剂检测中的应用及进展

液相色谱-质谱联用技术已越来越广泛地应用在兴奋剂的检测中,其中包括对各类小分子兴奋剂和肽类激素等的检测。本文综述了近年来液相色谱-质谱联用在兴奋剂检测中的筛选、确证和定量方面的应用及进展情况,并讨论了相关的检测标准。     

Rogue athletes and recombinant DNA technology: challenges for doping control

The quest for athletic excellence holds no limit for some athletes, and the advances in recombinant DNA technology have handed these athletes the ultimate doping weapons: recombinant proteins and gene doping. Some detection methods are now available for several recombinant proteins that are commercially available as pharmaceuticals and being abused by dopers. However, researchers are struggling to come up with efficient detection methods in preparation for the imminent threat of gene doping, expected in the 2008 Olympics. This Forum article presents the main detection strategies for recombinant proteins and the forthcoming detection strategies for gene doping as well as the prime analytical challenges facing them.     

Procedures for monitoring recombinant erythropoietin and analogues in doping control

The present report summarizes the main analytical strategies developed to identify the presence of recombinant erythropoietin (EPO) administered as a doping agent. Indirect evidence is based on the analysis of blood parameters (haemoglobin, haematocrit, reticulocytes, macrocytes, etc.) and serum markers (concentration of EPO and serum transferrin receptors, etc.). The problem of intertechnique comparison for reliable results evaluation is emphasized, especially for serum markers. Charge differences between isoforms of recombinant EPO and native urinary EPO are the grounds for the isoelectric focusing–double blotting–chemiluminescence detection method presently approved for doping control. Works addressing its advantages and limitations are presented and commented on. The chemical bases of the differential detection are highlighted and some future approaches for detection are also presented. The appearance and detectability of EPO analogues and mimetics susceptible for abuse are also addressed.     

Current strategies of blood doping detection

During the last 30 years, the artificial increase of red blood cell volume (“blood doping”) has changed the level of performance in all endurance sports. Many doping scandals have shown the extent of the problem. The detection of blood doping relies on two different approaches: the direct detection of exogenous manipulating substances (erythropoietic stimulants) or red cells (homologous transfusion) and the indirect detection, where not the doping substance or technique itself, but its effect on certain biomarkers is measured. Whereas direct detection using standard laboratory procedures such as isoelectric focusing can identify erythropoietic stimulants, homologous blood transfusion is identified through mismatches in minor blood group antigens by flow cytometry. Indirect methods such as the athlete biological passport are the only means to detect autologous transfusion and may also be used for the detection of erythropoietic stimulants or homologous transfusion. New techniques to unmask blood doping include the use of high-throughput ‘omics’ technologies (proteomics/metabolomics) and the combination of different biomarkers with the help of mathematical approaches. Future strategies should aim at improving the use of the available data and resources by applying pattern recognition algorithms to recognize suspicious athletes and, on the basis of these findings, use the appropriate testing method. Different types of information should be combined in the quest for a forensic approach to anti-doping.     

PCR-based detection of gene transfer vectors: application to gene doping surveillance

Athletes who illicitly use drugs to enhance their athletic performance are at risk of being banned from sports competitions. Consequently, some athletes may seek new doping methods that they expect to be capable of circumventing detection. With advances in gene transfer vector design and therapeutic gene transfer, and demonstrations of safety and therapeutic benefit in humans, there is an increased probability of the pursuit of gene doping by athletes. In anticipation of the potential for gene doping, assays have been established to directly detect complementary DNA of genes that are top candidates for use in doping, as well as vector control elements. The development of molecular assays that are capable of exposing gene doping in sports can serve as a deterrent and may also identify athletes who have illicitly used gene transfer for performance enhancement. PCR-based methods to detect foreign DNA with high reliability, sensitivity, and specificity include TaqMan real-time PCR, nested PCR, and internal threshold control PCR.     

New analytical strategies in studying drug metabolism

Identification and elucidation of the structures of metabolites play major roles in drug discovery and in the development of pharmaceutical compounds. These studies are also important in toxicology or doping control with either pharmaceuticals or illicit drugs. This review focuses on: new analytical strategies used to identify potential metabolites in biological matrices with and without radiolabeled drugs; use of software for metabolite profiling; interpretation of product spectra; profiling of reactive metabolites; development of new approaches for generation of metabolites; and detection of metabolites with increased sensitivity and simplicity. Most of the new strategies involve mass spectrometry (MS) combined with liquid chromatography (LC).     

毛细管区带电泳用于多种类兴奋剂的同时快速分离检测

建立了一种同时分离检测包括利尿剂、蛋白同化剂、β-阻断剂、麻醉剂、β2-激动剂、刺激剂等6类8种兴奋剂的毛细管区带电泳-紫外检测法。优化的色谱条件为：以50 mmol/L甲酸铵-氨水(pH 7.8)缓冲液为运行液,于3 kPa下进样10 s,分离电压为20 kV,检测波长为214 nm。在此条件下,8种兴奋剂在7 min内实现了快速的基线分离。在相应的浓度范围内,8种组分的浓度与峰高呈良好的线性关系,检出限达为0.2~0.7 μg/mL。该方法快速,分析成本低,无污染,非常适用于多种类兴奋剂的同时快速检测。     

Unintentional doping in GaN

The optimisation of GaN-based electronic and optoelectronic devices requires control over the doping of the material. However, device performance, particular for lateral transport electronic devices, is degraded by the presence of unintentional doping, which for heteroepitaxial GaN layers grown in the polar (0001) orientation is mainly confined to a layer adjacent to the GaN/substrate interface. The use of scanning capacitance microscopy (SCM) has demonstrated that this layer forms due to the high rate of incorporation of gas phase impurities, primarily oxygen, during the early stages of growth, when N-rich semi-polar facets are often present. The presence of such facets leads to additional unintentional doping when defect density reduction strategies involving a three-dimensional growth phase (such as epitaxial lateral overgrowth) are employed. Many semi-polar epitaxial layers, on the other hand, exhibit significant unintentional doping throughout their thickness, except when a three-dimensional growth phase is introduced to aid in defect density reduction resulting in the presence of (0001) and non-polar facets which incorporate less dopant. Non-polar epitaxial samples exhibit behaviour more similar to (0001)-oriented material, but oxygen diffusion from the sapphire substrate along prismatic stacking faults also locally affects the extent of the unintentional doping in this case.     

Rationally Design of Near Infrared Light Responsive Micro‐Photoelectrodes for In Vivo Sensing of Neurotransmitter Molecules in Mouse Brain†

The accurate quantification of neurotransmitter molecules is an indispensable means to reveal the physiological mechanisms of neuro movement in molecular level. However, existing detection strategies cannot fully meet practical needs, and the on‐site and in vivo detection of neurotransmitters in brain remains a great challenge. Here, we report the development of a near infrared light responsive photoelectrochemical (PEC) detection method for in vivo quantification of neurotransmitter dopamine in mouse brain. Under guidance of density function theory calculations, a combination strategy of non‐metal cation doping and defect engineering is introduced to rationally design the micro‐photoelectrodes with excellent biocompatibility and stability and implements the in vivo PEC detection of dopamine in mouse brain. It opens up a new way for the accurate in vivo detection of biomolecules and allows researchers to make novel inquiries for long‐standing questions in a new way.