慢性重型乙型肝炎患者血浆蛋白质组差异表达的初步研究

目的探讨慢性重型乙型肝炎患者血浆蛋白质组变化,为研究慢性重型乙型肝炎的发病机制及治疗提供新的线索。方法收集慢性重型乙型肝炎患者和健康对照者血浆各20份,去除血浆中高丰度蛋白。应用双向凝胶电泳（2-DE）技术构建两组研究对象血浆蛋白2-DE图谱,经Image Master差异分析软件进行分析,寻找差异蛋白质点。用电喷雾离子串联质谱（ESI-MS-MS）对重要的差异蛋白质点进行鉴定。结果经过样品预处理后,血浆中白蛋白和免疫球蛋白IgG含量大大减少,低丰度蛋白得到较好的富集。正常健康组蛋白质点287个,慢性重型乙型肝炎组蛋白质点297个,筛选出2倍以上的差异蛋白质点21个。鉴定出8个慢性重型乙型肝炎相关蛋白质,其中上调的蛋白质有α1-抗胰蛋白酶,载脂蛋白E;下调的蛋白质有补体因子B、CD5抗原样蛋白、纤维蛋白原B链、α-1B糖蛋白、载脂蛋白AI、触珠蛋白。结论成功鉴定了8个慢性重型乙型肝炎相关蛋白质。这些差异蛋白质功能涉及能量代谢、脂蛋白代谢、蛋白质分泌、补体旁路活化途径等过程。对这些重要蛋白质的结构和功能的进一步研究,对于进一步揭示慢性重型乙型肝炎发病分子机制及寻找治疗靶标可能具有重要的意义。     

载脂蛋白E基因与额颞叶痴呆临床特征的关系

目的探讨载脂蛋白E(Apo E)基因与额颞叶痴呆(FTD)临床特征的关系。方法选取FTD患者37例,年龄相仿的正常对照51例,应用多态性分析法检测Apo E基因型。应用χ2检验/Fisher确切概率法分析APOEε4基因携带和FTD临床特征的关系。结果 FTD患者的精神行为症状,淡漠占81%,易激惹、脱抑制各为54%,激越/攻击57%。10例ε4携带患者均以行为异常为首发症状,70%以记忆力减退为首发症状;而27例非ε4携带患者中,16例以行为异常为首发症状,11例以语言障碍为首发症状,12例有记忆减退的症状。ε4携带者出现幻觉(80%,P=0.001)、妄想(70%,P=0.017)、抑郁/心境恶劣(60%,P=0.012)的比例较高,差异具有统计学意义。结论 FTD患者的精神行为症状,以淡漠、易激惹、脱抑制、激越/攻击的比例较高;ε4携带者的首发症状以行为异常最常见;ε4携带者出现幻觉、妄想、抑郁/心境恶劣比例较高。     

蛋白质组研究在肾脏病领域的应用

基因组载有生物体的全部遗传信息 ,人类基因组计划的完成是人类发展史上重要的里程碑之一 ,标志着生命科学已由研究结构基因组进展至功能基因组时代[1] 。基因是遗传信息的携带者 ,而蛋白质是生物功能真正的执行者 ,其表达水平、存在方式及相互作用等与生物功能直接相关。蛋白质与mRNA之间并不存在严格的线性对应关系 ,Anderson等[2 ]认为两者的相关系数仅为 0 4 8,因此mRNA水平的研究不能替代蛋白质水平的研究。基因组基本上是固定不变的 ,蛋白质组是动态变化的 ,具有时空性和可调节性 ,它反映基因表达的时间、数量以及蛋白质翻译后修…     

额颞叶痴呆-TDP的病理和临床研究进展

额颞叶退行性变(FTLD)是一组以行为、人格改变和失语为特征性表现的疾病,包括3种临床综合征:行为变异额颞叶型痴呆(bvFTD)即狭义的额颞叶痴呆,额颞叶痴呆额叶型、语义性痴呆(SD),进行性非流利性失语(PNFA)〔1〕。FTLD是一个广义的范畴,临床表现、病理学特征、遗传学病因上都具有异质性。目前许多研究正在寻找其遗传学特征、病理学表现与临床症状之间的关联。     

双向凝胶电泳和二维色谱技术检测大肠癌蛋白质谱的意义

目的利用双向凝胶电泳和二维色谱技术分析Dukes B期大肠癌肿瘤与癌旁组织蛋白质差异,为大肠癌的早期诊断、生物学治疗寻找新的靶点。方法①双向凝胶电泳技术和质谱技术鉴定大肠癌肿瘤与癌旁组织的差异蛋白质组;②二维色谱技术和质谱技术鉴定大肠癌肿瘤与癌旁组织的差异蛋白质组。结果在大肠癌肿瘤组织与癌旁组织差异蛋白中两种技术路线有13个蛋白得到一致的鉴定结果,其中在肿瘤组织中表达上调的有7个,表达下调的有6个。结论两种技术路线共同鉴定出的差异蛋白质是有意义的,对研究大肠癌的发病、转移及复发机制提供了依据和方向。     

伴进行性延髓麻痹的额颞叶变性1例

病例:女,60岁,大专文化,商业单位主管,因"言语表达障碍、行为异常1年余"入院。入院1年前家属发现患者说话词不达意,无故欣快,行为幼稚,并伴记忆力下降,遂就诊当地医院,行头颅CT检查未见异常,而后行头颅磁共振成     

双向凝胶电泳与质谱技术分析Dukes B期大肠癌肿瘤与癌旁组织的差异蛋白质组学

目的研究Dukes B期大肠癌肿瘤与癌旁组织差异蛋白质组学。方法①应用双向凝胶电泳(2-DE)和质谱技术对大肠癌肿瘤与癌旁组织分离分析。②在鉴定出具有差异蛋白质中,选出2个蛋白质烯醇化酶(Enolase)、血红蛋白(Hemoglob in)行蛋白印记实验,验证2-DE结果。结果通过2-DE和质谱技术对大肠癌肿瘤与癌旁组织进行分离分析,21个蛋白质点在组织中的表达量差异显著。通过免疫蛋白印记实验进一步证明2-DE结果的准确性。结论 Dukes B期大肠癌肿瘤与癌旁组织蛋白质具有差异性,其中14个蛋白质点在肿瘤组织上调,7个下调。     

燃煤污染型砷中毒肝损伤患者血清差异蛋白质的筛选

目的 对比分析燃煤污染型砷中毒肝损伤患者和健康人血清中差异表达的蛋白,筛选与燃煤污染型砷中毒致肝损伤发生密切相关的血清蛋白质.方法 6例血清标本来自于贵州省兴仁县交乐病区燃煤型砷中毒肝损伤患者及病区健康人(对照组),采用双向凝胶电泳(2-DE)分离血清中总蛋白,硝酸银染色后经图像分析识别差异表达的蛋白质,基质辅助激光解吸电离飞行时问质谱(MALDI-TOF-MS)鉴定差异表达的蛋白质点,寻找与燃煤污染型砷中毒致肝损伤有关的蛋白质.结果成功地建立了血清2-DE图谱.差异蛋白分析显示,砷中毒肝损伤组平均蛋白点数为(824±31)个,对照组为(782±42)个,两组血清2-DE图谱的匹配率为94.9%(782/824).筛选出两组间的差异蛋白点49个,对其中表达量差异3倍以上的30个蛋白点进行MALDI-TOF-MS分析,鉴定出相关蛋白10个.与对照组比较,其中α2-巨球蛋白、B细胞受体相关蛋白31、细胞角蛋白1、载脂蛋白A-I在砷中毒肝损伤组表达上调,触珠蛋白、α2-HS-糖蛋白、细胞外信号调节激酶4、锌指蛋白323、ZAP-70、SP40表达下调.结论成功地建立了分辨率高、重复性较好的燃煤污染型砷中毒肝损伤患者血清2-DE图谱,并筛选出一些与健康人血清中差异表达的蛋白质,这些蛋白质能否作为燃煤污染型砷中毒肝损伤诊断的血清标志物还有待于进一步验证.     

心血管系统蛋白质组的研究进展

继基因组之后蛋白质组研究已成为当前生物医学研究的热点,检查蛋白质组的变化可深入理解基因组分析不能明确的细胞和分子机制,蛋白质组技术的发展促进了心血管疾病在分子机制上更全面的研究.本文概述了蛋白质组技术及其在心血管系统的研究进展.     

燃煤污染型砷中毒肝损伤患者血清差异蛋白质的筛选

Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.     

中国大陆日本血吸虫地理株间成虫蛋白质组分的差异

目的分离、鉴定中国大陆日本血吸虫不同地理株间成虫差异表达蛋白。方法分别收集、制备日本血吸虫不同地理株成虫可溶性蛋白,经固相pH梯度双向凝胶电泳分离,凝胶银染,并用PDQuest8.0凝胶图像分析软件进行比较分析,筛选出差异表达蛋白点并采用基质辅助激光解析离子飞行时间质谱仪进行鉴定。结果湖南株、江西株和江苏株雄虫分别检出698±10、650±19、629±23个蛋白点,雌虫分别检出670±12、682±22、625±28个蛋白点,约90%蛋白点分子量处于20～90kD范围内,蛋白等电点在5～8之间。不同地理株雌虫与雄虫分别两两比较,雄虫发现14个差异点,雌虫发现18个差异点。对其中7个雄虫差异表达蛋白及3个雌虫差异表达蛋白经MALDI-TOF-MS鉴定分析,获得其肽质量指纹图谱、等电点、分子量等相关信息。7个雄虫差异表达蛋白分别为:SJCHGC00821蛋白、SJCHGC00475蛋白、3-磷酸甘油醛脱氢酶、谷胱甘肽转移酶片段、谷胱甘肽-S-转移酶、D-核酮糖-5-磷酸-3-表异构酶、G蛋白α亚基AgGq1,3个雌虫表达蛋白分别为:预测蛋白、GAF型传感器信号转导组氨酸激酶、组织蛋白酶B样半胱氨酸蛋白酶前体。结论中国大陆日本血吸虫不同地理株虫体间蛋白质存在差异,部分蛋白表达差异与虫体对吡喹酮敏感性密切相关。     

Neuropsychiatric characteristics of PiB-negative subcortical vascular dementia versus behavioral variant frontotemporal dementia

BackgroundNeuropsychiatric symptoms of subcortical vascular dementia (SVaD) are mainly associated with damage to frontal-subcortical circuits and may be similar to symptoms of behavioral variant frontotemporal dementia (bvFTD). The aim of this study was to determine whether the neuropsychiatric manifestations of the Pittsburgh compound B (PiB)-negative SVaD and bvFTD groups differ.MethodsWe compared the Caregiver-Administered Neuropsychiatry Inventory (CGA-NPI) between 48 patients with PiB(−) SVaD and 31 patients with bvFTD. A stepwise logistic regression was applied to determine the best model to predict SVaD.ResultsThe SVaD group showed a higher frequency of depression, whereas the bvFTD group had a higher frequency of elation, aberrant motor behavior and appetite/eating disorders. Regarding NPI subscores, the bvFTD group had greater severity of elation, apathy, disinhibition, aberrant motor behavior and appetite/eating disorders, whereas SVaD did not have significantly higher subscores in any domains. The most predictive models that tend to find suggestions of SVaD, as opposed to bvFTD, are as follows: (1) the presence of depression and the absence of appetite/eating disorders, (2) higher NPI subscores of depression and lower NPI subscores of irritability and aberrant motor behavior.ConclusionApart from apathy, SVaD differed from bvFTD in that negative symptoms were more common in SVaD than bvFTD, whereas positive symptoms were predominant in bvFTD compared to SVaD.     

MALDI-TOF-MS分析新疆食管鳞状细胞癌的蛋白组学

目的:分析新疆食管鳞状细胞癌和食管正常上皮细胞的差异表达蛋白.方法:运用激光捕获显微切割技术(LCM)分别获取食管鳞状癌细胞和食管正常上皮细胞,应用二维凝胶电泳技术(2-DE)分离纯化细胞,Imagemaster 2D软件比较分析两者电泳图谱的差异,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定分析两者表达的差异蛋白.结果:建立了食管鳞状癌细胞和食管正常上皮细胞的2-DE图谱,获得43个差异蛋白点,通过质谱鉴定出17种蛋白,其中15种蛋白如Trangelin2、HSP27、S100A11、GSTP等在食管鳞状癌细胞中表达明显增高,2种蛋白如SCCA1,在食管鳞状癌细胞中表达明显降低.结论:提示17种差异蛋白可能与食管鳞状细胞癌的发生和发展有关,为筛选食管鳞状细胞癌的特异性分子标志物奠定基础.     

Proteomic analysis of hippocampus in the rat

Objective To analyze the protein expression in the rat hippocampus by the proteomic approach. Methods Proteins from hippocampal tissue homogenates of the rat were separated by two-dimensional gel electrophoresis (2-DE), and stained with colloidal Coomassie blue to produce a high-resolution map of the rat hippocampus proteome. Selected proteins from this map were digested with trypsin, and the resulting tryptic peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) The mass spectrometric data were used to identify the proteins through searches of the NCBI protein sequence database. Results 37 prominent proteins with various functional characteristics were identified. The identified brain protein classes covered metabolism enzymes, cytoskeleton proteins, heat shock proteins, antioxidant proteins,signalling proteins, proteasome-related proteins, neuron-specific proteins and glial-associated proteins. Furthermore,3 hypothetical proteins, unknown proteins so far only proposed from their nucleic acid structure, were identified.Conclusion This study provides the first unbiased characterization of proteins of the rat hippocampus and will be used for future studies of differential protein expression in rat models of neurological disorders.     

Proteomic analysis of gastric cancer and immunoblot validation of potential biomarkers

AIM: To search for and validate differentially expressed proteins in patients with gastric adenocarcinoma.METHODS: We used two-dimensional gel electrophoresis and mass spectrometry to search for differentially expressed proteins in patients with gastric adenocarcinoma. A set of proteins was validated with immunoblotting.RESULTS: We identified 30 different proteins involved in various biological processes: metabolism, development, death, response to stress, cell cycle, cell communication, transport, and cell motility. Eight proteins were chosen for further validation by immunoblotting. Our results show that gastrokine-1, 39S ribosomal protein L12 (mitochondrial precursor), plasma cell-induced resident endoplasmic reticulum protein, and glutathione S-transferase mu 3 were significantly underexpressed in gastric adenocarcinoma relative to adjacent non-tumor tissue samples. On the other hand, septin-2, ubiquitin-conjugating enzyme E2 N, and transaldolase were significantly overexpressed. Translationally controlled tumor protein was shown to be differentially expressed only in patients with cancer of the gastric cardia/esophageal border.CONCLUSION: This work presents a set of possible diagnostic biomarkers, validated for the first time. It might contribute to the efforts of understanding gastric cancer carcinogenesis.     

Perspective: Proteomic approach to detect biomarkers of human growth hormone

Several serum biomarkers for recombinant human growth hormone (rhGH) have been established, however, none alone or in combination have generate a specific, sensitive, and reproducible ‘kit’ for the detection of rhGH abuse. Thus, the search for additional GH specific biomarkers continues. In this review, we focus on the use of proteomics in general and two-dimensional electrophoresis (2-DE) in particular for the discovery of new GH induced serum biomarkers. Also, we review some of the protocols involved in 2-DE. Finally, the possibility of tissues other than blood for biomarker discovery is discussed.     

Can proteomics lead to the discovery of real biomarkers for HCC?

The development of proteomics technologies has lead to a great deal of effort being focused on the identification of biomarkers for cancers. Although many papers have reported candidate biomarkers for hepatocellular carcinomas (HCCs) in particular, so far none of these candidate biomarkers have been used either for diagnosis or therapy intreating patients. The question remains: Can proteomics identify real biomarkers for HCCs?     

Differential protein expression profiling of myocardial tissue in a mouse model of hypertrophic cardiomyopathy

Hypertrophic cardiomyopathy (HCM) is a genetic disorder caused by mutations in genes encoding sarcomere proteins. The mechanisms involved in the development of cardiac hypertrophy and heart failure remain poorly understood. Global proteomic profiling was used to study the cardiac proteome of mice predisposed to developing HCM. Hearts from three groups of mice (n = 3 hearts per group) were studied: non-transgenic (NTG) and cardiac-specific transgenic models over-expressing either the normal (TnI^WT) or a mutant cardiac troponin I gene (Gly203Ser; TnI^G203S). Two-dimensional gel electrophoresis (2-DE) coupled with tandem mass spectrometry was used to identify proteins. Image analysis was performed using Progenesis SameSpots. A total of 34 proteins with at least a twofold change in the TnI^G203S mouse model were identified. Alterations were detected in components involved in energy production, Ca²⁺ handling, and cardiomyocyte structure. Expression level changes in cytoskeletal and contractile proteins were well represented in the study, including the intermediate filament protein desmin, which was further investigated in two additional physiological and pathological settings, i.e., exercise treatment, and severe heart failure in a novel double-mutant TnI-203/MHC-403 model of HCM. This study highlights the potential role of tissue proteomic profiling for mapping proteins, which may be critical in cardiac dysfunction and progression to heart failure in HCM.     

刚地弓形虫蛋白质组研究进展

随着基因组学的发展和完善,在基因组学的基础上,蛋白质组学作为一门新兴的前沿科学已经显示出了其巨大的发展前景.将蛋白质组学技术应用于弓形虫研究,在蛋白质水平上全面认识弓形虫的生理、病理等过程是目前弓形虫研究领域的热点之一.该文就弓形虫蛋白质组的研究进展进行综述,以期从蛋白质组学研究的角度为弓形虫研究提供新的思路.