液相色谱/离子阱质谱法研究何首乌中糖苷类化合物

采用液相色谱／离子阱质谱（HPLC／IT-MS）联用技术研究何首乌中糖苷类化合物,并对大黄素甲醚糖苷和大黄素糖苷的分子离子或准分子离子的离子化机理进行探讨。实验采用反相C18色谱柱,二元线性梯度洗脱,分离出12个主要成分。利用质谱的诱导碰撞解离技术获得碎片裂解信息,结合文献鉴定出9种糖苷的化学结构,其中顺式-2,3,5,4＇-四羟基二苯乙烯-2-O-β-D-毗喃葡萄糖苷、决明酮-8—O-（6＇-O-乙酰基）-β—D-吡喃葡萄糖苷和大黄素甲醚-8-O-（6＇-O-乙酰基）-β-D-吡喃葡萄糖苷为首次鉴定。     

FATTY ACID DYNAMICS IN THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE): IMPLICATIONS FOR P HYSIOLOGICAL ECOLOGY1,2,3

The growth dry weight, fatty acid weight and fatty acid composition of two clones of Thalassiosira pseudonana Hasle & Heimdal were measured under several growth conditions. Determinations of total cellular fatty acids were made using chemical ionization mass spectrometry. Both clones had the same fatty acids, dominated by C14:0, C16:0, C16:1, C16:3, C16:4, C18:4 and C20:5, though in different relative amounts. Fatty acids typically represented 5–10% the dry weight of a cell during log Phase growth and up to 22% during stationary Phase. The C16 fatty acids of both clones changed as the cultures aged, though much more markedly in the Sargasso Sea done (13–1) than in the estuarine one (3H). The C16:0 and C16:1 acids of both clones declined sharply in the dark and were replenished in the light. Cells maintained in constant illumination, but with no cell division. produced large quantities of these acids. Cells of done 13–1 treated with polychlorinated biphenyl (PCB) initially grew more slowly than control cells, weighed more, and had higher relative amounts of C16:0 and C16:1. Fatty acid studies may provide useful indicators of ecologically important energy reserves and membrane adaptations in the algae.     

Deuterium marking of chemical emissions: detectability and fitness consequences of a novel technique for insect behavioural studies

Behavioural studies often need to employ marking techniques to identify and track individuals. Marks ideally have no influence on behaviour or fitness of marked individuals or on the organisms with which they interact. A newly developed internal marking technique uses deuterium to manipulate the chemical emissions of bethylid wasps that are parasitoids of lepidopteran larvae. Wasps are deuterated indirectly by injecting deuterium oxide (heavy water) into the host on which they feed as larvae. Adult wasps emit a volatile chemical under stressful conditions and the emissions of deuterated and undeuterated wasps can be clearly and readily distinguished by measuring the ratio of deuterated/undeuterated molecules using real‐time mass spectrometry. Deuterated female wasps are, however, detectably disadvantaged in dyadic contests for host resources, a component of female fitness. In this study, we evaluated potential side‐effects of this deuterium marking technique on three further correlates of female Goniozus legneri Gordh (Hymenoptera: Bethylidae) fitness: longevity, lifetime fecundity, and offspring developmental mortality. We found that deuterium enrichment has no overall adverse effects on these parameters. We further showed that injecting hosts with different concentrations of deuterium results in distinguishably different volatile emissions of adult wasps, such that at least three individuals can be simultaneously identified by their chemical signatures. We also showed that deuterium marking remains effective beyond a wasp's death: deuterated and undeuterated wasps remain readily distinguishable after storage. We discuss potential applications of deuterium marking for the laboratory and field study of chemical communication, dispersal, and mating structure in parasitoid wasps and other insects.     

Method for determination of methadone in exhaled breath collected from subjects undergoing methadone maintenance treatment

At present drugs of abuse testing using exhaled breath as specimen is only possible for alcohol. However, we recently discovered that using modern liquid chromatography–mass spectrometry technique amphetamine and methamphetamine is detectable in exhaled breath following intake in drug addicts. We therefore undertook to develop a method for determination of methadone in exhaled breath from patients undergoing methadone maintenance treatment. Exhaled breath was collected from 13 patients after intake of the daily methadone dose. The compounds were trapped by filtering the air through a C18 modified silica surface. After elution of any trapped methadone the extract was analysed by a combined liquid chromatography–tandem mass spectrometry method. Recovery of trapped methadone from the filter surface was 96%, no significant matrix effect was observed, and the quantification using methadone-d3 as an internal standard was accurate (<10% bias) and precise (coefficient of variation 1.6–2.0%). Methadone was indisputably identified by means of the mass spectrometry technique in exhaled breath samples from all 13 patients. Identification was based on monitoring two product ions in selected reaction monitoring mode with correct relative ratio (±20%) and correct retention time. Excretion rates ranged from 0.39 to 78 ng/min. No methadone was detected in 10 control subjects. This finding confirms that breath testing is a new possibility for drugs of abuse testing. Collection of exhaled breath specimen is likely to be more convenient and safe as compared to other matrices presently in use.     

Quantitative proteomics suggests decrease in the secretogranin‐1 cerebrospinal fluid levels during the disease course of multiple sclerosis

Multiple sclerosis (MS) is a chronic inflammatory disease of the CNS with unknown cause. Proteins with different abundance in the cerebrospinal fluid (CSF) from relapsing‐remitting MS (RRMS) patients and neurological controls could give novel insight to the MS pathogenesis and be used to improve diagnosis, predict prognosis and disease course, and guide in therapy decisions. We combined iTRAQ labeling and Orbitrap mass spectrometry to discover proteins with different CSF abundance between six RRMS patients and 18 neurological disease controls. From 777 quantified proteins seven were selected as biomarker candidates, namely chitinase‐3‐like protein 1, secretogranin‐1 (Sg1), cerebellin‐1, neuroserpin, cell surface glycoprotein MUC18, testican‐2 and glutamate receptor 4. An independent sample set of 13 early‐MS patients, 13 RRMS patients and 13 neurological controls was used in a multiple reaction monitoring verification study. We found the intracellular calcium binding protein Sg1 to be increased in early‐MS patients compared to RRMS and neurological controls. Sg1 should be included in further studies to elucidate its role in the early phases of MS pathogenesis and its potential as a biomarker for this disease.     

Proteomics for the detection of indirect markers of steroids treatment in bovine muscle

Despite the ban by the European Union, anabolic steroids might still be illicitly employed in bovine meat production. The surveillance of misuse of such potentially harmful molecules is necessary to guarantee consumers’ health. Analytical methods for drug residue control are based on LC‐MS/MS, but their efficacy can be hindered due to undetectable residual concentrations as a result of low‐dosage treatments. Screening methods based on the recognition of indirect biological effects of growth promoters’ administration, such as the alteration of protein expression, can improve the efficacy of surveillance. The present study was aimed at identifying modifications in the muscle protein expression pattern between bulls treated with an ear implant (Revalor‐XS®) containing trenbolone acetate (200 mg) and estradiol (40 mg), and untreated animals. The analysis of skeletal muscle was carried out using a tandem mass tags shotgun proteomics approach. We defined 28 candidate protein markers with a significantly altered expression induced by steroids administration. A subset of 18 candidate markers was validated by SRM and allowed to build a predictive model based on partial least square discriminant analysis. Our findings confirm the effectiveness of the proteomics approach as potential tool to overcome analytical limitations of drug residue monitoring.     

孕妇孕期营养指导及体质量控制对妊娠结局的影响

目的:对孕妇进行孕期营养指导及体质量控制,探究其对妊娠结局的影响。方法:将2011年2月-2014年2月期间我院收治的孕产妇150例纳入本次研究,所有研究对象随机均衡分为研究组和对照组,每组各75例。对照组孕产妇仅行常规妇科检查,研究组孕产妇在医护人员指导下行孕期营养指导和体质量控制,比较两组分娩方式,妊娠过程状况及围产儿情况。结果:研究组剖宫产率为17.33%低于对照组的57.33%,组间差异存在统计学意义(P0.05)。研究组贫血、羊水过少、产后出血率均低于对照组,差异有统计学意义(P0.05)。研究组胎龄及巨大儿及胎儿窘迫率均低于对照组,差异有统计学意义(P0.05)。结论:孕产妇在孕期进行营养膳食指导,控制体质量在合理范围内,可以改善孕妇妊娠结局,降低妊娠过程中并发症发病率,提高围产儿状况,保证孕妇和婴儿健康。     

Deciphering preferential interactions within supramolecular protein complexes: the proteasome case

In eukaryotic cells, intracellular protein breakdown is mainly performed by the ubiquitin–proteasome system. Proteasomes are supramolecular protein complexes formed by the association of multiple sub-complexes and interacting proteins. Therefore, they exhibit a very high heterogeneity whose function is still not well understood. Here, using a newly developed method based on the combination of affinity purification and protein correlation profiling associated with high-resolution mass spectrometry, we comprehensively characterized proteasome heterogeneity and identified previously unknown preferential associations within proteasome sub-complexes. In particular, we showed for the first time that the two main proteasome subtypes, standard proteasome and immunoproteasome, interact with a different subset of important regulators. This trend was observed in very diverse human cell types and was confirmed by changing the relative proportions of both 20S proteasome forms using interferon-γ. The new method developed here constitutes an innovative and powerful strategy that could be broadly applied for unraveling the dynamic and heterogeneous nature of other biologically relevant supramolecular protein complexes.