Comparative analysis of the liver and plasma proteomes as a novel and powerful strategy for hepatocellular carcinoma biomarker discovery

The extraordinary developments made in the past decade in proteomic technologies, in particular in mass spectrometry, have enabled investigators to consider designing studies to search for diagnostic and therapeutic biomarkers by scanning complex proteome samples. We developed a method based on extensive fractionation of intact proteins, to comprehensively and quantitatively profile the liver and plasma proteomes in health and disease. We have applied this method to samples collected from patients with early hepatocellular carcinoma (HCC) and from patients with liver cirrhosis as well as to samples collected from three mouse models of HCC. This method allowed for the identification of proteins that differ in expression levels in liver tissue or in plasma with disease progression from liver fibrosis, cirrhosis or steatohepatitis to HCC. The comparative analysis of the liver and plasma proteomes generated from human and mouse specimens, constitutes a novel and powerful strategy for HCC biomarker discovery.     

“3P”医学新概念

“3P”医学是将预警、预防以及针对不同患者的个性化治疗有机地结合为一体，被誉为21世纪医学发展的新方向，代表医学发展的终极目的和最高阶段。后基因组时代系统生物学的目的就是使卫生保健和医疗实践活动发生根本性转变，从而引导医学研究与实践进入“3P”医学的新时代。本文阐述了“3P”医学和系统生物学的基本概念以及它们对系统医学的作用。     

黄曲霉毒素B1诱发树鼩肝癌过程中的差异表达蛋白质分析及意义

目的比较黄曲霉毒素B1(AFB1)诱发的树鼩肝细胞癌生长不同阶段的蛋白质表达谱的改变,筛查在AFB1诱发的肝癌发生发展中起重要作用的关键蛋白质分子.方法用双向电泳(2-DE)及基质辅助激光解吸飞行时间串联质谱法(MALDI-TOF-MS/MS)对同一动物自身的癌发生前肝组织(A45W),癌发生后的肝癌组织(ACa)及未经AFB1处理的对照组动物肝组织(E45W)进行了相互比较.结果A45W组、ACa组、E45W组分别平均检测到(1 255±43)个蛋白点(n=3)、(1 198±50)个蛋白点(n=3),和(1 252±40)个蛋白点(n=3),以其中A45W组图谱为参考胶,ACa组、E45W组分别与其匹配的平均匹配点数为(818±39)个(n=3)和(777±45)个(n=3),平均匹配率分别为66%和62%.癌与癌前比(ACa vs A45W),相差2倍以上的上调点和下调点分别为108个(包括癌特异点)和75个;癌前与未经AFB1处理的对照组比(A45W vs E45W),相差2倍以上的上调点有78个,下调点有21个.质谱可鉴定出不均一核糖核蛋白A1、不均一核糖核酸核蛋白A2/B1、peroxiredoxin 1、peroxiredoxin 2、膜联蛋白4、肌球蛋白α链等26种胶内差异蛋白质.结论在AFB1诱发的树鼩肝癌发生不同阶段有明显的差异性蛋白表达谱,为进一步对人肝癌蛋白质组的研究提供了线索.     

蛋白质飞行质谱技术在严重急性呼吸综合征(SARS)早期诊断中的应用价值(英文)

目的探索蛋白质飞行质谱技术在严重急性呼吸综合征(SARS)早期诊断中的应用价值。方法用表面加强激光解吸电离质谱技术检测急性SARS组和对照组的血清,首先建立诊断模型,然后单盲进行模型验证。结果通过检测急性SARS组(74例)和对照组(147例)的血清,发现了早期SARS患者血清中质谱峰(M/Z)为3939.08、4137.71、8136.64、11514.20的4种特异蛋白指纹标志物,并建立诊断模型,此模型敏感性98.6%(73/74),特异性94.6%(139/147)。结论早期SARS患者有特异的蛋白指纹图谱,利用表面加强激光解吸电离质谱技术检测SARS患者或疑似SARS患者血清,可更好地协助临床进行SARS的早期诊治。     

Proteomics of diphtheria toxoid vaccines reveals multiple proteins that are immunogenic and may contribute to protection of humans against Corynebacterium diphtheriae

Introduced for mass immunization in the 1920s, vaccines against diphtheria are among the oldest and safest vaccines known. The basic principle of their production is the inactivation of purified diphtheria toxin by formaldehyde cross-linking, which converts the potentially fatal toxin in a completely harmless protein aggregate, which is still immunogenic. Since in addition to diphtheria toxin also other proteins may be secreted by Corynebacterium diphtheriae during cultivation, we assumed that diphtheria toxoid might not be the only component present in the vaccine. To address this question, we established a protocol to reverse formaldehyde cross-linking and carried out mass spectrometric analyses. Different secreted, membrane-associated and cytoplasmic proteins of C. diphtheriae were detected in several vaccine preparations from across the world. Based on these results, bioinformatics and Western blot analyses were applied to characterize if these proteins are immunogenic and may therefore support protection against C. diphtheriae. In frame of this study, we could show that the C. diphtheriae toxoid vaccines induce antibodies against different C. diphtheriae proteins and against diphtheria toxin secreted by Corynebacterium ulcerans, an emerging pathogen which is outnumbering C. diphtheriae as cause of diphtheria-like illness in Western Europe.     

乳腺癌血清蛋白质组与尿代谢物组协同分析研究

目的 研究血清蛋白质组和尿代谢物组协同分析乳腺癌诊断方法。方法 用双向电泳、高效液相色谱和液质联用技术对7份正常人与14份乳腺癌患者血清蛋白质组和尿代谢物组进行分析，从中寻找差异蛋白和代谢组模式差异。结果 与正常人相比，乳腺癌患者血清中谷胱甘肽S-转移酶M5呈高表达状态，尿中乳清酸核苷、1-甲酰化腺苷、S-腺苷-L-蛋氨酸及N^2-甲酰化鸟苷等4种核苷代谢物组成模式异常。结论 乳腺癌患者体液内化学物质与正常人相比具有特异性模式差异。经肿瘤蛋白质组与代谢物组协同研究，有望建立肿瘤早期诊断新方法，进行肿瘤发病机理研究。     

蛋白质组学分析在淋巴瘤中的应用进展

蛋白质组学分析已经广泛应用于多种肿瘤的临床研究,用于发现可以应用于临床的潜在生物学标志物.近年,与淋巴瘤相关的蛋白质组学分析研究也正在进行中,主要涉及淋巴瘤的发病机制、诊断、分型、治疗及预测预后等方面.为了使相关临床工作者更好地了解蛋白质组学分析及其应用价值,笔者拟就蛋白质组学分析在淋巴瘤中的应用进展进行综述.     

New serological biomarkers of inflammatory bowel disease

Serological biomarkers in inflammatory bowel disease （IBD） are a rapidly expanding list of non-invasive tests for objective assessments of disease activity, early diagnosis, prognosis evaluation and surveillance. This review summarizes both old and new biomarkers in IBD, but focuses on the development and characterization of new serological biomarkers （identifi ed since 2007）. These include fi ve new anti-glycan antibodies, anti-chitobioside IgA （ACCA）, anti-laminaribioside IgG （ALCA）, anti-manobioside IgG （AMCA）, and antibodies against chemically synthesized （∑） two major oligomannose epitopes, Man α-1,3 Man α-1,2 Man （∑Man3） and Man α-1,3 Man α-1,2 Man α-1,2 Man （∑Man4）. These new biomarkers serve as valuable complementary tools to existing biomarkers not only in differentiating Crohn＇s disease （CD）, ulcerative colitis （UC）, normal and other non-IBD gut diseases, but also in predicting disease involvement （ileum vs colon）, IBD risk （as subclinical biomarkers）, and disease course （risk of complication and surgery）. Interestingly, the prevalence of the antiglycan antibodies, including anti-Saccharomyces cerevisiae antibodies （ASCA）, ALCA and AMCA, was found to be associated with single nucleotide polymorphisms （SNPs） of IBD susceptible genes such as NOD2/CARD15, NOD1/CARD4, toll-likereceptors （TLR） 2 and 4, and β-defensin-1. Further-more, a gene dosage effect was observed： anti-glycan positivity became more frequent as the number of NOD2/CARD15 SNPS increased. Other new serum/ plasma IBD biomarkers reviewed include ubiquitination factor E4A （UBE4A）, CXCL16 （a chemokine）, resistin, and apolipoprotein A-IV. This review also discusses the most recent studies in IBD biomarker discovery by the application of new technologies such as proteomics, fourier transform near-infrared spectroscopy, and multiplex enzyme-linked immunosorbent assay （ELISA）＇s （with an emphasis on cytokine/chemokine profiling）. Finally, the prospects o     

铁必复颗粒治疗老年性聋的临床疗效与分子机制研究

目的观察铁必复颗粒治疗老年性聋的临床疗效,通过实验探讨该方对肾虚耳聋治疗作用的分子机制。方法老年性聋患者136例,随机分两组,铁必复颗粒组70例（138耳）,西药对照组66例（129耳）,前者接受铁必复颗粒冲服,后者口服尼莫地平等。实验研究选用Wistar大鼠120只,随机分为两组。正常对照大鼠组,喂标准饲料16W;缺铁大鼠组,采用缺铁饲料喂养8W后,再分成肾虚耳聋组、铁必复颗粒组,肾虚耳聋组继续喂以缺铁饲料8W,铁必复颗粒治疗组喂以添加铁必复颗粒的缺铁饲料8W。取大鼠耳蜗膜性组织进行蛋白质组学分析,比较各组蛋白质表达差异。结果老年性聋患者铁必复颗粒组显效21耳,有效42耳,有效率45．65％;西药对照组有效10耳,有效率7．75％,前者有效率高于后者,两组疗效差异有统计学意义（P〈0．01）。实验研究表明,肌动蛋白、肌球蛋白、细胞骨架蛋白等在铁必复颗粒治疗组耳蜗膜性组织中高表达,与正常组和肾虚耳聋组相比,蛋白表达都有上调。铁必复颗粒组出现特异性高表达46kDaprotein、48kDaprotein等尚未命名的蛋白。结论铁必复颗粒治疗老年性聋有效,其作用机制可能与调控耳蜗肌动、肌球、细胞骨架蛋白等的表达水平有关。