化学发光分析技术在生命科学研究中的一些应用

生命过程是一个错综复杂的过程，人类对生命现象的探索由来己久。随着现代检测诊断技术，如X射线照影，B超，电子显微和CT技术等的出现，人类对生命活动的认识以及疾病的诊断取得了突飞猛进的发展。在应用这些方法观察生命过程中的宏观现象的同时，人们也把目光投向隐藏在这些宏观现象背后的微观过程，渴望从分子级别上认识生命的各种过程，即对参与生命过程的物质进行定性、定量和活性测试。核酸和蛋白质是在生命活动中最重要的两类物质，而要分析它们在生命体系中的活动并非易事，因为生命体系大都十分复杂。所以针对不同的分析对象建立有效的核酸和蛋白质分析方法已成为生命科学研究中的前沿和热门课题。目前世界上许多研究机构和商业企业在这方面投入大量的人力和财力，以期建立有效的方法和推出相应的商业试剂和仪器。纵观这方面的发展现状，我们不难发现许多当前流行的核酸和蛋白质分析方法中采用了化学发光体系。本文就化学发光法在分析核酸和蛋白质方面的发展现状做一些介绍。     

基于专利分析的化学发光免疫分析技术全球发展趋势研究

基于Innography平台,本文对化学发光免疫分析技术全球专利进行检索和统计分析,从专利权人和国家地区分布、专利质量分析和专利文本聚类分析以及专利侵权风险等角度进行深入解读。分析结果表明,全球化学发光免疫分析研究总体呈上升趋势,专利申请主要集中在美国、日本、德国、中国,美国综合实力最强,高价值专利量最多,最大的两家公司-罗氏和雅培各有一个庞大的专利家族对自己的核心技术进行保护。文本聚类的结果揭示:1)肿瘤标志物、心脏标志物的检测是热点应用领域; 2)化学发光微粒子免疫分析、磁性微粒、固相载体等技术是近年来的热点技术。最后对中国化学发光免疫分析技术的发展提出了建议。     

生物活性物质的电致化学发光检测

电致化学发光是通过电极上直接或间接发生的电化学反应而产生的一种化学发光,因此电致化学发光检测是在化学发光和电化学基础上发展起来的一种新的分析技术。电致化学发光检测技术不但保留了化学发光分析和电化学分析固有的优点,同时还具有其自身的优点,如所发生的化学发光反应易于控制;方法更灵敏,更具有选择性;可以获得更多的化学信息;扩大了化学发光方法可检测的范围;更易于与现代分离技术联用。生物体中很多生物活性物质具有电活性,因此用现代电化学技术研究其电化学行为具有重要的理论意义和实际应用价值。生物体中的生物活性物质通常浓度很低,并且成分复杂,因此分离检测生物体中生物活性物质非常困难。由于电致化学发光检测具有灵敏度高,选择性好的特点,无疑是检测生物体中生物活性物质的强有力工具,如果它能与HPLC、CE及FIA等现代分离技术相结合,将表现出更加强大的生命力。     

特邀编者按

我国的化学发光研究起步于20世纪80年代，章竹君教授于1976年在《化学通报》发表了第一篇介绍化学发光研究的文章。随后，福州大学、陕西师范大学、中国科技大学等单位相继开展了化学发光基础理论研究、仪器研制、试剂合成和人才的培养。1990年代以前，我国的化学发光研究处于起步状态，1990年代以前     

钌(Ⅱ)配合物化学发光反应及其分析应用研究进展

近几年来，钌(Ⅱ)配合物化学发光反应及其分析应用研究进展较快，本文对钌(Ⅱ)配合物化学发光反应新体系、分析应用新方法、与分离技术联用、试剂衍生与固定化等方面的研究进行了评述。     

化学发光传感器研究进展

本文综述了1999年以后国内外化学发光传感器研究进展，其内容包括化学发光传感器的设计、酶基化学发光生物传感器、化学发光组织传感器、免疫化学发光传感器、催化型化学发光气体传感器、分子印迹型化学发光传感器、化学发光反应同时作为分子识别反应的化学发光传感器，并对今后的发展趋势进行展望。     

化学发光免疫分析

本文综述了化学发光免疫分析的基本原理及应用。根据标记物的不同，化学发光免疫分析主要分为三大类：化学发光免疫分析，化学发光酶免疫分析和电化学发光免疫分析。引用文献89篇。     

生物大分子的化学发光成像研究

本文介绍了三种化学发光成像法在人血清结合珠蛋白分型电泳的快速检测方面的应用。这三种方法都是基于鲁米诺一过氧化氢发光体系。使用免疫化学发光成像法，直接化学发光成像法和增强化学发光成像法，能够提高对结合珠蛋白检测的灵敏度，获得更宽的线性范围。我们使用这三种方法完成了对结合珠蛋白的快速、高灵敏、高选择性的检测。     

偶合反应化学发光分析

偶合反应化学发光分析是扩大化学发光分析应用范围的一种有效途径。本文主要从酶联化学发光分析、无机偶合反应化学发光分析、光化学反应一化学发光分析和电化学反应一化学发光分析四个方面对偶合反应化学发光分析的研究状况进行了评述。     

基于纳米材料的化学发光传感器研究

本文合成了多种纳米材料,在此基础上设计了乙醇、乙醛、氨、三甲基胺等化学发光传感器。并对传感器的灵敏度、选择性、使用寿命等参数进行了评价．证明了这类传感器具有良好的传感性能,是一类有前途的化学发光传感器。     

A simple,sensitive, non-stimulated photon counting system for detection of superoxide anion in whole blood

A simple, sensitive, non-stimulated assay was developed to measure the superoxide anion concentration in whole blood, using an ultra-sensitive chemiluminescence (CL) analyzer, and lucignin amplification. The assay system can be performed without leukocyte isolation or stimulant administration. The blood CL levels of healthy males (362.8±337.7 counts/10 sec) were not different from those of females (335±308.7 counts/10 sec) (p=0.64), whereas the CL levels in whole blood in patients with acute pancreatitis (2522±2014 counts/10 sec) were significantly higher than those of healthy controls (p<0.001). This assay system may be valuable in the future for quantitative measurement of reactive oxygen species in various disorders.     

The effect of surgery and anesthetic agents on granulocyte-chemiluminescence in whole blood

Summary The effect of anesthesia and major abdominal surgery on zymosan-induced chemiluminescence (CL) of neutrophil granulocytes was evaluated. CL was measured in diluted whole blood taken at distinct intervals within the perioperative period. In addition, blood samples from healthy volunteers were supplemented with ether and halothane to investigate the in vitro effect of these agents. The phagocytosis-induced CL was not found to be depressed by anesthesia and surgery. Only at supranarcotic concentrations was CL reduced. Surgery and anesthesia, therefore, do not appear to impair this defense system significantly under the conditions of this investigation.     

The effect of surgery and anesthetic agents on granulocyte-chemiluminescence in whole blood

The effect of anesthesia and major abdominal surgery on zymosan-induced chemiluminescence (CL) of neutrophil granulocytes was evaluated. CL was measured in diluted whole blood taken at distinct intervals within the perioperative period. In addition, blood samples from healthy volunteers were supplemented with ether and halothane to investigate the in vitro effect of these agents. The phagocytosis-induced CL was not found to be depressed by anesthesia and surgery. Only at supranarcotic concentrations was CL reduced. Surgery and anesthesia, therefore, do not appear to impair this defense system significantly under the conditions of this investigation.     

Requirements for leukocyte transmigration via the transmembrane chemokine CX3CL1

The surface-expressed transmembrane CX3C chemokine ligand 1 (CX3CL1/fractalkine) induces firm adhesion of leukocytes expressing its receptor CX3CR1. After shedding by the disintegrins and metalloproteinases (ADAM) 10 and 17, CX3CL1 also acts as soluble leukocyte chemoattractant. Here, we demonstrate that transmembrane CX3CL1 expressed on both endothelial and epithelial cells induces leukocyte transmigration. To investigate the underlying mechanism, we generated CX3CR1 variants lacking the intracellular aspartate-arginine-tyrosine (DRY) motif or the intracellular C-terminus which led to a defect in intracellular calcium response and impaired ligand uptake, respectively. While both variants effectively mediated firm cell adhesion, they failed to induce transmigration and rather mediated retention of leukocytes on the CX3CL1-expressing cell layer. Targeting of ADAM10 led to increased adhesion but reduced transmigration in response to transmembrane CX3CL1, while transmigration towards soluble CX3CL1 was not affected. Thus, transmembrane CX3CL1 mediates leukocyte transmigration via the DRY motif and C-terminus of CX3CR1 and the activity of ADAM10.     

Volume-dependent regulation of the respiratory burst of activated human neutrophils

The effect of incubation medium osmolality on the respiratory burst of human neutrophils was studied using luminol-dependent chemiluminescence (CL) as an indicator of burst activity. Neutrophils were stimulated with N-formyl-Met-Leu-Phe (FMLP), phorbol-12-myristate-13-acetate (PMA), the calcium ionophore A23187, thermoaggregated IgG (IgG_n), and opsonized zymosan (OZ). It was shown that increasing the osmolality of the incubation medium from 320 up to 420 mosM decreased the A231870 and OZ-induced CL responses by 90%. Under the same conditions PMA-, FMLP- and IgG_n-induced CL responses were decreased by 40–60%. A decrease of osmolality to 200 mosM resulted in a 2–3 fold decrease of the A23187-, PMA- and FMLP-induced CL and in a 60–80% increase of OZ- and IgG_n-induced CL. It is suggested that osmolality-mediated alteration of cell volume is an important mechanism for regulating neutrophil activity.     

金银藤挥发油成分的气-质联用分析与化学计量学解析

目的 定性、定量分析金银藤挥发油成分.方法 采用气相色谱-质谱进行检测,通过化学计量学解析法对二雏色谱/质谱数据进行解析,得到各组分的纯色谱曲线和质谱,从而实现对金银藤挥发油成分的定性、定量分析,并结合程序升温保留指数来鉴定化合物.结果 金银藤的挥发油鉴定了39个成分,定性组分含量占金银藤挥发油成分总含量的94.54%.结论 全银藤挥发油的主要成分为十六烷酸（45.86%）、9,12-十八碳二烯政乙酯（39.02%）、十四烷酸（1.8%）、十五烷酸（1.07%）、十八烷酸（1.02%）.     

Lipid peroxidation,low-level chemiluminescence and regulation of secretion in the mammary gland

In mammary explants of lactating mice, changes in the intensity of chemiluminescence (CL) were observed after the addition to the incubation medium of hormones and mediators that are involved in the regulation of secretion: oxytocin, acetylcholine, epinephrine and norepinephrine. A 15-min period of treatment with oxytocin, epinephrine or norepinephrine changed the level of thiobarbituric acid-reactive substances (TBARS). Two mammary explants, one of which was treated with oxytocin, acetylcholine, epinephrine or norepinephrine, were found to interact even when separated by a quartz glass wall. Analysis of the level of TBARS formation in these two explants showed that the observed interactions might be connected with light emission resulting from lipid peroxidation (LP) processes. The possible role of LP and low-level CL in the regulation of mammary gland secretion is discussed.     

Alloxan acts as a prooxidant only under reducing conditions: influence of melatonin

Depending on the availability of suitable reducing agents, alloxan can be either a prooxidant or an antioxidant. Alloxan and its reduced derivative, dialuric acid, act as a redox couple, driven by reduced glutathione (GSH) or L-cysteine, generating in vitro in the presence of oxygen, both superoxide radical and hydrogen peroxide. The production of superoxide radicals was shown by the appearance of lucigenin chemiluminescence (CL) as well as by the generation of formazan from nitroblue tetrazolium (NBT). The lucigenin CL as well as the NBT reduction was inhibited by superoxide dismutase and partially by catalase. Melatonin inhibited alloxan-mediated CL. In contrast, in the absence of reducing agents, alloxan is a scavenger of superoxide radicals formed by other reactions. Because of the high content of reducing compounds in the cell (e.g. glutathione), it is suggested that alloxan acts in vivo mainly as a generator of reactive oxygen species. Received 9 November 1998; received after revision 15 January 1999; accepted 15 January 1999     

γ-Glutamyltranspeptidases: sequence, structure, biochemical properties, and biotechnological applications

γ-Glutamyltranspeptidases (γ-GTs) are ubiquitous enzymes that catalyze the hydrolysis of γ-glutamyl bonds in glutathione and glutamine and the transfer of the released γ-glutamyl group to amino acids or short peptides. These enzymes are involved in glutathione metabolism and play critical roles in antioxidant defense, detoxification, and inflammation processes. Moreover, γ-GTs have been recently found to be involved in many physiological disorders, such as Parkinson's disease and diabetes. In this review, the main biochemical and structural properties of γ-GTs isolated from different sources, as well as their conformational stability and mechanism of catalysis, are described and examined with the aim of contributing to the discussion on their structure-function relationships. Possible applications of γ-glutamyltranspeptidases in different fields of biotechnology and medicine are also discussed.