质谱在金属抗癌药物与蛋白质相互作用研究中的应用

研究细胞毒性金属抗肿瘤药物与蛋白质的相互作用,以及这种相互作用对药物的细胞摄入、转运、代谢和生物利用度的影响,对金属抗癌药物的结构设计和优化,提高药物的抗癌活性,降低毒副作用具有重要意义。基于软电离技术的电喷雾质谱和基质辅助激光解析电离质谱能够在分析检测过程中很好的保留金属抗癌药物与蛋白质的共价(配位)结合,获得药物与蛋白质结合位点的信息。同时,质谱分析还具有灵敏度高,所需样品量少,耗时短以及适用于分析复杂生物样品等优点,已成为研究金属抗癌药物与蛋白质相互作用最强有力的工具,在为药物发现提供大量化学、生物信息的同时,也极大地促进了质谱技术自身的发展。本文将结合我们在金属抗癌药物相互作用组学研究中取得的最新进展,系统地总结、评述Bottom-up和Top-down质谱分析方法在铂、钌类金属抗癌药物与蛋白质相互作用研究中的发展动态,并分析这一前沿交叉领域未来的发展趋势。     

亲和超滤质谱技术在中药活性成分筛选中的研究进展

亲和超滤质谱技术是20世纪90年代中期发展起来的一种快速、简单、有效的药物小分子发现模式。该技术利用配体与受体之间特异性结合,通过超滤装置快速筛选活性小分子化合物,再结合液相色谱 质谱联用技术（LC-MS）,鉴定活性成分结构。亲和超滤质谱技术集药物活性成分筛选、结构鉴定于一体,尤其适用于从复杂体系中筛选潜在的活性小分子化合物。近年来,针对中药发挥药理作用具有多组分、多靶点的重要特点,亲和超滤质谱技术已被广泛用于从中药提取物中筛选与特定蛋白靶点相结合的小分子活性物质,对阐明中药药效的物质基础和以活性成分作为先导化合物的新药开发具有重要意义,是对传统药物发现方法的有利补充。本工作综述了该技术在中药活性成分筛选中的原理、特点、应用进展,以及对未来的展望。     

质谱在新药研究中应用的回顾

回顾了质谱在新药研究中的一些应用 ,包括新药合成、化学结构确证、质量控制以及药物代谢物的鉴定等。综述了基质辅助激光解吸电离 \飞行时间质谱 ( Matrix-assistant laser desorption ionization time-of-flight mass spectrometry,MALDI/ TOFMS)、电喷雾电离质谱 ( Electrode spray ionization m ass spectrome-try,ESI-MS)等仪器及其联用技术在生物大分子药物研究 ,包括蛋白质组学、肽图谱、氨基酸序列、核苷酸分析等方面的进展 ,展望了质谱在未来生命科学中广阔的应用前景。     

基质辅助激光解吸电离-飞行时间质谱在高分子研究中的应用

在质谱学的发展过程中,如何获得大分子物质的质谱信息一直是最具有挑战性的课题。在过去的20年里,大分子的质谱研究取得了很大进展,发展出快原子轰击（FAB）、液质联用（LC-MS）、基质辅助激光解吸电离-飞行时间质谱（MALDI-TOF）以及质谱/质谱串联（MALDI-TOF/TOF）等技术,并在生物大分子的结构测定、高聚物的结构及端基测定、共聚物的序列结构及组成等方面对高分子科学的发展作出了突出贡献。本文结合具体实例着重介绍了MALDI-TOF MS在大分子分析中的技术优势及应用     

质谱电喷雾电离源研究新进展

自1984年约翰芬恩(John Fenn)发明一种软电离离子源,即电喷雾电离源(ESI)以来,促进了质谱技术在大分子分析领域,特别是生物大分子领域的广泛使用。ESI源的低取样效率由ESI源的离子化效率和传输效率的构成,制约着ESI源质谱灵敏度的提高,因此研究者们一直努力改进ESI源的性能,以提高取样效率、改善ESI源质谱灵敏度。本研究介绍了ESI源的原理,并评述了近年来ESI源研究的新进展和应用。     

生物质谱在核糖核酸领域的应用

综述了近十年来生物质谱用于分析核糖核酸的应用进展。概述了电喷雾电离质谱(ESI-MS)、基质辅助激光解析电离质谱(MALDI-MS)的原理。论述了生物质谱用于分析生物大分子具有准确、快速、灵敏度高等优点。总结了质谱用于单核苷酸多态性分型分析(SNP genotyping);对短的串联重复序列(STR)分析;基因缺陷而导致的疾病诊断;对寡核苷酸片段的序列分析等4个方面的研究成果。讨论了应用ESI-MS分析寡核苷酸形成的非共价复合物,包括研究寡核苷酸的高级结构、寡核苷酸与蛋白质间形成的复合物、寡核苷酸与其它配体小分子间相互作用。提出了存在的问题,并展望了生物质谱未来的发展方向。     

现代生物质谱及其应用

80年代以来,有机质谱获得突破性进展,其应用已跨入了生物大分子的研究领域,并迅速形成了一个新的分支学科生长点——生物质谱学。已经诞生的生物质谱计已在生命科学的研究中发挥着非常重要的作用。本文主要介绍基质辅助激光解吸电离/飞行时间质谱(MALDI/TOF MS)的构造、原理、功能及主要应用。     

基于纳米毛细管的非变性质谱分析

非变性质谱利用电喷雾电离质谱技术从非变性缓冲溶液中研究自然折叠状态下的蛋白质、核酸及其复合物,是大型蛋白质组装体、蛋白质-药物复合物、核酸-配体复合物的结构、组成及其相互作用解析的重要工具。生化缓冲溶液中的非挥发性盐能提供足够的离子强度和适宜的pH值,以维持蛋白质、核酸分子折叠和结合状态,但通常与电喷雾电离质谱不兼容。基于纳米毛细管的电喷雾电离具备强大的基质耐受能力,为直接从生理或近生理缓冲溶液中完成蛋白质及其复合物的高效离子化提供了可能,极大地促进了近年来非变性质谱的应用研究。本文综述了纳米毛细管的制备与表征方法及其尺寸和界面性质导致的电离行为特性,回顾了近10年来纳米毛细管在非变性质谱中的应用研究进展。     

高分辨快原子轰击质谱法及其在生物医学分析中的应用

快原子轰击质谱经过十几年的发展，已成为分析极杜强、难挥发和热不稳定化合物囊常用的质谱方法，然而用快原子轰击质话进行精确质量测定的应用和研究开展还不很广泛．本文简要介绍了高分辨快原子轰击腐语法的方法和特点，并概述了高分辨快原子轰击质谱在生物医学分析几个主要方面（生物小分子、生物大分子、药物代谢产物和临床病理研究等）中的应用．     

MALDI-TOF MS在聚合物分析中的应用研究进展

近年来,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)作为一种新型的软电离质谱技术,具有有快速、准确、灵敏度高、检测范围广等特点,在化学、生物,医疗卫生等众多领域得到广泛研究和应用。本文着重介绍了MALDI-TOF-MS的工作原理,基质的选择,该技术在聚合物分析中的应用进展,以及存在的问题,并对其发展前景作出展望。     

斑蝥素及其药理作用研究进展

斑蝥属鞘翅目芫菁科昆虫,是我国最早发现的具有抗肿瘤作用的中药材之一,其药用成分是一种单萜类有毒物质——斑蝥素。本为就斑蝥素的资源分布、生化特性、在虫体中的分布及药理学作用进行了综述,为人类进一步认识和利用斑蝥素提供了理论基础。     

Light microscopy and scanning electron microscopy: Implications for authentication of misidentified herbal drugs

Plant‐based drugs have reached remarkable acceptability as therapeutic remedy for various diseases due to the adverse effects of contemporary medicines. This increasing popularity of herbal drugs leads to a growing herbal market for the development of plant‐based drugs, nutraceuticals, pharmaceuticals and cosmeceuticals. Herbal drug adulteration is a complex problem which currently has undeniable consequences on health and nutrition. Ambiguities in nomenclature, misidentification and resemblance of colour and texture of the crude herbal drugs are the major causes of adulteration. Three different species commercially marketed under the same trade name Halion are Lepidium apetalum, Asparagus officinalis, and Lepidium didymum. The genuine source of Halion is Lepidium apetalum, which is authenticated by using basic and advanced taxonomic techniques. Morphology, anatomy and palynology of the misidentified sources were done using light and scanning electron microscopic techniques for authentication. This study may help to set microscopic techniques as a tool to achieve quality and standardization of the genuine source of the herbal drug. Phytochemical analysis and biological screening is needed for the further establishment of authenticity and quality of herbal drugs.     

Proteomics analysis provides novel biomarkers and therapeutic target candidates in the treatment of the Huang-Pu-Tong-Qiao formula in an AD rat model

Background: Huang-Pu-Tong-Qiao formula (HPTQ), a traditional Chinese herbal formula, has a variety of pharmacological effects. It has been used to treat Alzheimer’s disease (AD) for decades. This study aimed to screen differentially expressed proteins in the hippocampus of AD model rats treated with HPTQ. Proteomic studies of the effects of HPTQ on AD are key to understanding the therapeutic mechanisms of HPTQ and identifying potential therapeutic targets. Methods: We hence used the isobaric tags for relative and absolute quantification (ITRAQ) approach to investigate the differentially expressed proteins in the hippocampus of AD model rats before and after HPTQ administration and to identify the potential therapeutic target proteins of HPTQ. In this study, the learning and memory abilities of AD rats were examined by the Morris water maze test. After HPTQ administration, the differentially expressed proteins in the hippocampus of AD rats were quantified and analyzed in silico. Furthermore, western blotting was used to verify the expression of related proteins. Results: The Morris water maze results showed that HPTQ could improve the learning and memory ability of AD model rats. The proteomics analysis results showed that 57 proteins were differentially expressed, of which 35 were up-regulated and 22 were down-regulated. Bioinformatics analysis indicated that proteins with altered expression after HPTQ treatment were involved in several biological processes that have the potential to exert neuroprotective effects. These included promoting the translation of ribosomes, improving the deposition of amyloid-beta (Aβ), regulating autophagy, regulating neuronal synaptic function and plasticity, and alleviating oxidative stress. Conclusion: In conclusion, we identified several potential therapeutic target proteins and related mechanistic pathways of HPTQ in the treatment of AD, laying the foundation for further investigation of the therapeutic effects of HPTQ.     

Scutellarin alleviates complete freund’s adjuvant-induced rheumatoid arthritis in mice by regulating the Keap1/Nrf2/HO-1 pathway

Scutellarin (SCU) is a herbal flavonoid glucuronide with multiple pharmacological activities, including anti-oxidant, anti-inflammation, vascular relaxation, anti-platelet, and myocardial protection. However, the effect of SCU on complete Freund’s adjuvant (CFA)-induced rheumatoid arthritis (RA) had not been studied. In this study, we investigated the beneficial effects of SCU in the CFA-induced RA mice model and the anti-arthritic activity was evaluated by paw edema. Enzyme-linked immunosorbent assay (ELISA) was carried out to evaluate the plasma levels of immunoglobulin (Ig)G, IgE, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, receptor activator of nuclear factor-κB ligand (RANKL), and osteoprotegerin (OPG). Histological slides were prepared from the harvested paws of mice to determine the pathological changes in the joints. The proportions of T helper type 1 (Th1) and T helper type 2 (Th2) cells of CD4+ T lymphocyte subsets were analyzed by flow cytometry. The expression of Kelch-like ECH-associated protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) was analyzed using real-time quantitative PCR (RT-qPCR) and western blotting assays. The present study demonstrated that SCU prevented CFA-induced RA, and inhibited the expression of inflammation factors, IgG, IgE, TNF-α, IL-1β, and IL-6. While SCU also reduced the RANKL level, it increased OPG expression in RA mice. The Th1/Th2 ratio was significantly lower in mice treated with SCU. Additionally, HO-1 expression was reduced while the expression of Keap1 and Nrf2 was elevated following SCU treatment. Results provide preliminary evidence to employ SCU in arthritis treatment which might be related to the regulation of Th1/Th2 balance and the Keap1/Nrf2/HO-1 pathway.     

通过RRLC-Q-TOF MS和UPLC-QQQ MS分析原人参三醇型皂苷在人肠道菌群中的代谢产物

通过高分离度快速液相色谱-四极杆飞行时间质谱(RRLC-Q-TOF MS)和超高效液相色谱-三重四极杆质谱(UPLC-QQQ MS)法对原人参三醇型皂苷Re、Rg 1、Rg 2、Rh 1、Rf、F 1、R 1在人肠道菌群中的转化产物进行定性、定量分析,确定原人参三醇型皂苷的代谢产物、转化途径和60 h时的转化率。结果表明,人参皂苷Re的转化产物为人参皂苷Rg 1、Rg 2、Rh 1、F 1和PPT,转化率为91%;人参皂苷Rg 1的转化产物为人参皂苷Rh 1、F 1和PPT,转化率为80%;人参皂苷Rg 2的转化产物为人参皂苷Rh 1和PPT,转化率为73%;人参皂苷Rh 1和F 1主要通过PPT代谢,转化率分别为82%和81%;人参皂苷Rf的转化产物为人参皂苷Rh 1和PPT,转化率为89%;三七皂苷R 1的转化产物为人参皂苷Rg 1、R 2、Rh 1和PPT,转化率为79%。原人参三醇型皂苷类成分可被人肠道菌群代谢,主要通过丢失糖残基形成转化产物,而次级皂苷和苷元是人参在体内发挥药理作用的物质基础。     

Authentication of herbal drug Tukhm‐e‐balango (Lallemantia royleana Benth.) using microscopic,pharmacognostic, and phytochemical characterization

The study is aimed to provide a comprehensive account on authentication of herbal drug named as Tukhm‐e‐balango (Lallemantia royleana Benth.) from the seeds of Ocimum basilicum by using microscopic, pharmacognostic, and phytochemical characterization. The crude medicinal plants and their parts are often adulterated or substituted in market due to improper identification by the consumers while among herbal plant sellers, taxonomic confusion is caused due to morphological similarities of the plant parts and lack of a standard identification system.In microscopy, both herbarium and fresh specimens were studied using qualitative and quantitative morphological characteristics of leaves, seeds, and pollen. For pharmacognosy, solubility, fluorescence, and physicochemical characterizers were analyzed whereas a total phenolic and flavonoids contents was determined in addition to DPPH radical scavenging activity. In current study, microscopic, pharmacognostic, and phytochemical characterization clearly differentiated L. royleana from O. basilicum. The major problem in herbal drug industry is caused due to confusion and controversy of certain synonyms used for more than one or two drugs. Sometimes, under the same common or local name, entirely different taxa are being sold in herbal markets. It is concluded that correct and proper identification of medicinal plants is very crucial to ensure the safety and efficacy of herbal medicines, as many medicinal plants are intentionally or unintentionally adulterated with similar species or varieties. In herbal market, the seeds of L. royleana are adulterated with seeds of O. basilicum due to their similar morphology.     

裂解-气相色谱-质谱法对中药材指

建立了离线裂解-气相色谱-质谱联用法(P-GC/MS)研究中药材指纹图谱的测定方法。中药材样品经粉碎,离线高温裂解,丙酮乙醇混合液萃取后,利用气相色谱-质谱联用法进行测定。质谱总离子流图充分反映中药材组分特性,并确定各峰组分的结构,为中药材鉴别及质量控制提供了一种很好的方法。该方法有效地改进了图谱的特征性和重现性,具有快速、简单、准确等特点,可用于中药材指纹图谱研究。     

中草药微波真空连续干燥机控制系统的设计

针对中草药微波真空连续干燥过程复杂,自动化程度要求高的问题,将PLC、触摸屏等自动化及人机工程技术引入到中草药微波真空连续干燥过程中,建立了由PLC、MP277(触摸屏)、物料输送机构(包括进料、出料)、调频器、制冷机组、集水器、真空泵、微波组件、电磁阀、温度传感器、湿度传感器、压力表和重量传感器等组成的中草药微波真空连续干燥控制系统。着重研究了中草药微波真空连续干燥机PLC自动控制系统的结构及原理,进行了PLC、触摸屏及重要传感器的选型,并编制了上位机及下位机控制程序。实际运行结果证实,该系统实现了中草药微波真空连续干燥过程的自动控制,提高了中草药微波真空连续干燥机的可靠性和自动化程度。     

<i>In vitro</i> study of emodin-induced nephrotoxicity in human renal glomerular endothelial cells on a microfluidic chip

Emodin is an effective component of rhubarb with positive pharmacological effects on human health. However, it is also toxic to different cells or tissues to varying degrees. The effects of emodin on glomerular endothelial cells (GECs) remain to be tested, and the documented works were always performed in vitro and hardly reflect the real physiological situation. To study the effects of emodin on GECs in a biomimetic environment, we utilized a microfluidic chip to assess the physiological reaction of human renal glomerular endothelial cells to various concentrations of emodin in this work. The results showed that emodin caused cytotoxicity, impaired glomerular filtration barrier integrity to macromolecules, and increased barrier permeability in a dose-dependent manner. With the increase in emodin concentration, the concentration of the pro-inflammatory cytokine tumor necrosis factor-α, interleukin (IL)-6, transforming growth factor-β1, and monocyte chemoattractant protein (MCP-1) increased while the production of inflammatory cytokine IL-6 first increased and then decreased with the increase in emodin concentration. Our findings shed new light on emodin-induced nephrotoxicity and provide insights for the application of microfluidic chip devices to reveal drug-cell interactions.     

An experimental study on four kinds of Chinese herbal medicines containing alkaloids using fluorescence microscope and microspectrometer

In order to find a useful method for uniting the identification and quality evaluation of Chinese herbal medicine, the techniques of fluorescence microscopy and microspectrometer were firstly applied to authenticate four kinds of herbal medicines, Caulis Sinomenii, Rhizoma Coptidis, Radix Sophorae Tonkinensis and Rhizoma Menispermi, as well as to measure the distribution of alkaloids in their cross-sections. The results showed that the fluorescence microscopic characteristics and the fluorescence emission spectra of the same tissues from the four kinds of Chinese herbal medicine were different, for example, the cortex of Radix Sophorae Tonkinensis emitted blue fluorescence instead of the yellow of Rhizoma Coptidis observed with emission filter of long-pass 397 nm, which could be effective to identify them. Furthermore, alkaloids, as active components of the above herbal medicines, were distributed in each part of herbal tissue but their fluorescence intensities in different parts of tissue were different. The results indicated that fluorescence microscope combined with microspectrometer could be a helpful method for identification and quality evaluation of Chinese herbal medicine.