应用SELDI-TOF-MS技术建立直肠癌筛选血清蛋白质指纹图谱模型

目的:建立直肠癌筛选血清蛋白质指纹图谱模型并初步验证. 方法:用表面加强激光解析电离飞行时间质谱技术(SELDI- TOF-MS)及WCX2蛋白芯片获得新发直肠癌、直肠息肉患者和正常人血清的蛋白质指纹图谱,用计算机软件进行比较分析,建立直肠癌的筛选模型,并对其进行了盲法验证．结果:直肠癌组与对照组共有26个蛋白质有显著性差异(P<0．05);以其中4个蛋白质生物标志物(质／荷比9 295,3 730,3 938和4 095)组建的筛选模型检测正确率为 96．8％(93／96),经盲法验证,其灵敏度为95．0％(38／40),特异性为93．4％(45／48)．结论:建立的血清蛋白质指纹图谱模型能够区分直肠癌与非直肠癌患者,SELDI-TOF-MS在直肠癌的诊断及肿瘤特异性蛋白质生物标志分子的筛选等方面具有一定价值．     

SELDI-TOF-MS技术在肝癌诊断和介入治疗评价中的价值

目的:建立肝癌血清学诊断模型,探讨评估SELDI-TOF-MS技术在肝癌诊断和介入治疗评价中的价值.方法:用弱阳离子交换芯片(CM10芯片)和表面增强激光解吸电离飞行时间质谱仪(surface-enhanced laser desorption ionization time-of-flight mass spectrometry,SELDI-TOF-MS)技术,测定60例肝癌患者和60例正常对照者的血清蛋白质指纹图谱,应用BiomarkerWizard统计软件比较肝癌组和正常对照组血清蛋白质表达的差异性,采用Biomarker Pattern软件分析数据建立肝癌诊断模型,比较介入治疗前后血清蛋白质指纹图谱的差异性.结果:在质荷比(M/Z)为2000-10000范围内,和正常血清比较,肝癌的差异峰有3个(M/Z为4182Da、5710Da、6992Da;P<0.01),4182Da和5710Da下调,6992Da上调.用这3个差异蛋白峰建立肝癌诊断模型,诊断肝癌的灵敏度为93.3%(28/30),特异度为90.0%(27/30),正确率为91.7%(55/60),约登指数为0.833.差异蛋白峰(M/Z4182Da)在介入术后1mo明显上调(P<0.05).结论:应用SELDI-TOF-MS技术进行肝癌血清蛋白质指纹图谱分析,建立肝癌诊断树模型,对肝癌的诊断有一定的价值;筛选出的差异蛋白峰对肝癌的介入治疗评估有一定的应用价值.     

类风湿关节炎患者血清蛋白质指纹图谱检测及其临床意义

目的初步探讨基于判别分析建立的血清蛋白质指纹图谱模型在类风湿关节炎(RA)诊断中的临床意义.方法用表面增强激光解吸电离飞行时间质谱技术及配套蛋白质芯片检测60例RA患者和31例健康人的血清蛋白质指纹图谱,并采用SPSS12.0软件判别分析处理数据和筛选标志物,以建立诊断模型.结果质荷比(m/z)为3 975 Da,6 433 Da和15 129 Da的3个蛋白质峰组合构建的诊断模型鉴别RA患者和健康人的敏感性为80.0%(48/60),特异性为80.6%(25/31).结论血清蛋白质指纹图谱在筛选RA患者血清中特异性蛋白标志物及其诊断方面具有一定价值.     

SELDI-TOF-MS在筛选肝癌高发家系与无癌家系血清蛋白差异中的应用

目的：比较肝癌高发家系患者和无癌家系患者血清蛋白质谱图,探寻两者间的血清蛋白差异。方法用表面增强激光解吸电离-飞行时间质谱技术（ SELDI-TOF-MS）及CM10蛋白芯片对25例肝癌高发家系患者和25例无癌家系患者血清样本进行分析,将获得的血清蛋白质指纹图谱,用计算机软件进行比较分析,建立高发家系患者和无癌家系患者的比较筛选模型。结果肝癌高发家系组与无癌家系组共有15个蛋白质差异有统计学意义。筛选出4个蛋白峰建立诊断模型的灵敏度为96.0％（24/25）,特异度为88.0％（22/25）,准确率为92.0％（46/50）。结论肝癌高发家系患者组和无癌家系患者组的血清存在一定的差异蛋白, SELDI技术在特异性蛋白生物标志分子的筛选等方面具有较好的临床应用前景。     

应用SELDI-TOF-MS技术建立SLE患者血清蛋白质指纹图谱筛选模型

目的筛选系统性红斑狼疮（SLE）血清蛋白质指纹图谱模型,并探讨其意义。方法用表面加强激光解析电离飞行时间质谱（SEIDI-TOF-MS）技术获得SLE、类风湿关节炎患者（RA组）、健康人群（对照组）血清中的蛋白质谱,用计算机软件比较分析,建立SLE血清蛋白质指纹图谱筛选模型,并验证该模型诊断SLE的灵敏度和特异性。结果SLE组与非SLE组共有11种蛋白质表达有显著性差异;以其中4个蛋白质生物标志物（5 739 Da、10 832 Da、4 650 Da、15 797 Da）组建的筛选模型,经双盲法验证其灵敏度为84.4%,特异性为90.3%。结论应用SEIDI-TOF-MS技术建立的血清蛋白质指纹图谱模型在SLE的早期诊断中具有一定价值。     

肝癌、肝硬化患者前期处理后腹水中差异蛋白质组学分析及意义

目的 分析肝癌、肝硬化患者腹水前期处理后的蛋白质组学变化,并探讨其意义.方法 采用表面加强激光解析电离飞行时间质谱技术（SELDI-TOF-MS技术）对27例肝癌（肝癌组）、46例肝硬化患者（肝硬化组）腹水前期处理后蛋白质进行检测,对蛋白质波谱进行分析.结果 肝癌组在7 852 Da、11 467 Da蛋白质表达量高于肝硬化组,在4 475 Da蛋白质表达量低于肝硬化组（P均＜0.05）.SELDI-TOF-MS技术检测7 852 Da、11 467 Da、4 475 Da区分肝癌和肝硬化灵敏度为81.48％ （22/27）,特异性为86.96％ （40/46）.结论 腹水经前期处理后,肝癌、肝硬化患者在7 852 Da、11 467 Da、4 475 Da蛋白质峰有显著差异,通过这三个蛋白质峰检测可以区分肝癌与肝硬化.     

SELDI技术筛选肺癌患者血清标志蛋白的临床价值

目的探讨表面增强激光解析电离飞行时间质谱（SELDI-TOF-MS）技术筛选肺癌患者血清标志蛋白的临床价值。方法用SELDI-TOF-MS技术、弱阳离子交换蛋白芯片,检测肺癌和肺良性病变患者的血清蛋白质质谱图;用Biomarker Pattern软件分析肺癌差异蛋白并初建其诊断模型,通过盲筛验证诊断模型。结果发现有统计学差异的蛋白峰20个,其中肺癌患者血清高表达蛋白质波峰14个,低表达蛋白质波峰6个;用质荷比2 090.77、2 503.31 Da的差异蛋白峰建立分类树模型,其诊断肺癌的灵敏度88%,特异度95%;盲筛验证灵敏度90%,特异度100%,粗符合率93.33%,Youden指数0.9。结论SELDI-TOF-MS技术筛选的肺癌血清差异性蛋白及分类树模型,诊断肺癌的灵敏度高、特异性好。     

应用蛋白指纹图谱技术筛选原发性胆汁性肝硬化患者血清特异性标志物

目的: 探讨蛋白指纹图谱技术筛选原发性胆汁性肝硬化(PBC)患者血清中可用于诊断的特异性标志物.方法: 采用弱阳离子纳米磁性微球捕获血清中的蛋白,ProteinChip PBSII-C型蛋白质芯片阅读仪检测绘制成蛋白指纹图谱.所有蛋白指纹图谱采用Biomarker Wizard 3.1分析之后用Biomarker Pattems Sofiware 5.0识别最终可能用于PBC诊断的蛋白标志物并优化组合建立诊断模型.结果: 在PBC患者和对照组之间找到69个差异蛋白峰(P<0.05).其中质荷比(m/z)为3445,4260,8133和16 290的蛋白峰建立PBC诊断模型.该诊断模型能很好地把PBC患者从其他肝脏疾病患者和正常人群中区分出来,其敏感性为93.3%,特异性为95.1%.经双盲实验验证,该模型对PBC诊断的敏感性为92.9%,特异性为82.4%. 结论: 采用纳米磁性微球与蛋白质芯片阅读仪联用的蛋白指纹图谱技术可以检测PBC患者血清中的特异性蛋白标志物,并建立敏感性和特异性均较高的PBC诊断模型.     

非小细胞肺癌患者血清蛋白质标记物的检测

目的 检测非小细胞肺癌患者(NSCLC)血清蛋白质,筛选特异的蛋白质标记物,构建用于NSCLC早期诊断的血清蛋白质指纹图谱模型.方法 应用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术检测235例血清标本的蛋白质质谱,并结合生物信息学方法(支持向量机)分析数据.结果 筛选出3个质荷比(m/z)位于6628,9191和11412的蛋白质标记物,构建NSCLC早期诊断模型.联合3种潜在蛋白质标记物,经留一法交叉验证,区分NSCLC和正常健康对照的敏感性为98%,特异性为96%.盲法验证显示,该模型诊断NSCLC的敏感性为96.56%,特异性为94.79%.结论 SELDI-TOF-MS结合支持向量机建立NSCLC血清蛋白质指纹图谱模型是早期诊断NSCLC的一种敏感性高、特异性强的新方法,值得进一步研究与应用.     

新疆汉族、维吾尔族及哈萨克族食管癌血清蛋白质指纹图谱

目的:探讨新疆主要民族之间食管癌患者血清蛋白质指纹图谱的差异.方法:用表面增强激光解吸电离飞行时间质谱技术(SELDI-TOF-MS)检测43例汉族、43例维吾尔族及41例哈萨克族食管癌血清蛋白质指纹图谱,所得结果用ZUCI-PDAS软件分析处理,筛选血清差异性蛋白质.结果:同民族间的血清蛋白质指纹图谱存在明显差异.汉族组与维吾尔族组中质荷比为4310.0109,8713.0142,7993.0223的3个蛋白质峰差异具有统计学意义(P<0.05);汉族组与哈萨克族食管癌组中质荷比为4310.0184, 8167.9277,8158.1117,13789.4864,8067.7056,4611.9098,7993.4422, 16146.8706的8个蛋白质峰差异具有统计学意义(P<0.05);维吾尔族组与哈萨克族组中质荷比分别为9161.7944,4611.6342,6649.6163, 4979.3807的4个蛋白质峰差异具有统计学意义(P<0.05);质荷比4310.0109的蛋白峰在汉族组中呈低表达,而在维吾尔族组和哈萨克族组中均呈高表达.在维吾尔族组和哈萨克族组之间该蛋白峰未见到有差异.结论:汉族、维吾尔族及哈萨克族食管癌患者之间血清蛋白质指纹图谱的差异具有统计学意义,在新疆地区建立不同民族食管癌患者血清蛋白质指纹图谱疾病诊断模型有一定的临床意义.     

New multi protein patterns differentiate liver fibrosis stages and hepatocellular carcinoma in chronic hepatitis C serum samples

~~New multi protein patterns differentiate liver fibrosis stages and hepatocellular carcinoma in chronic hepatitis C serum samples@Thomas Gbel$Klinik für Gastroenterologie,Hepatologie und Infektiologie,Heinrich-Heine-UniversitatDusseldorf,Moorenstr.e,D…     

表面增强的激光解析电离飞行时间质谱技术在肝细胞癌血清学诊断中的应用

目的:检测HBV感染携带者与肝细胞癌(HCC)患者血清蛋白质的差异性表达,以发现HCC的肿瘤诊断标志物.方法:用表面增强的激光解析电离飞行时间质谱(SELDI-TOF-MS)技术检测27例HCC患者,27例HBV感染携带者,25例健康对照血清中的蛋白质谱,并用Biomarker Patterns System 5．0软件分析,建立诊断模型．结果:检测HCC患者与HBV感染携带者。正常对照与HCC患者,正常对照与HBV感染携带者的差异性蛋白分子,据此构建分类模型,得到的灵敏度和特异度分别为93％,85％; 96％,96％;84％,89％．其中相对分子质量为8141 Da的蛋白峰在HCC组明显高于HBV感染组(p<10-5);相对分子质量为3448 Da的蛋白峰在HCC及HBV感染携带组表达均较正常组显著增高(P<10-5),而在HCC-HBV组无明显差异(P>0．05),提示其可能为HBV感染的一种标志蛋白;相对分子质量为777) Da的蛋白峰在3组中均有差异性表达．结论:SELDI蛋白芯片技术检测血清蛋白质谱法诊断HCC具有较高的灵敏度和特异度,操作简单快捷,临床应用前景广阔,为HCC诊断提供了新的血清学方法．     

肝细胞性肝癌患者介入治疗前后血清蛋白质组的变化

目的用表面增强激光解吸离子化飞行时间质谱（surface—enhanced laser desorption—ionization timeof—flightmass spectrometry,SELDI—TOF—MS）蛋白质芯片检测肝细胞性肝癌（简称肝癌）患者介入治疗前后血清蛋白质组的变化,初步探讨肝癌介入治疗的药物靶点。方法应用SELDI—TOF—MS蛋白质芯片检测50例未经治疗的肝癌患者、50例介入治疗的肝癌患者,获得弱阳离子交换芯片（weakcationicexchanger）蛋白表达图谱,Bio．MarkerWizard软件分析差异蛋白。结果未经治疗的肝癌患者和介入治疗的肝癌患者血清中检测到不同质荷比处有3个差异蛋白质峰,其蛋白质含量差异有统计学意义（P〈0．05）。数据库搜索这3个蛋白质峰,得到3种与之分子量最为接近的蛋白质。结论应用SELDI—TOF—MS筛选肝癌患者介入治疗的药物靶点快速、有效,检测到的3个差异蛋白质可能是肝癌患者介入治疗的药物靶点。     

蛋白芯片技术对肝细胞癌组织蛋白谱的建立及标志蛋白的筛选

目的:应用表面增强激光解吸附电离飞行时间质谱技术(SELDI-TOF-MS)对肝细胞癌组织的蛋白表达谱进行分析,从中筛选出标记蛋白.方法:采用CM10芯片及SELDI-TOF-MS技术对肝细胞癌组织26例和肝硬化组织18例进行蛋白指纹图谱检测分析;比较高、中、低分化肝细胞癌组织,不同TNM分期肿瘤以及AFP阴、阳性肝细胞癌组织的蛋白表达差异,所得结果均采用Biomarker Wizard软件进行分析;通过查询蛋白库,对特定分子质量所对应的标记蛋白进行初步确定.结果:肝细胞癌和肝硬化组织蛋白表达图谱之间存在16个稳定的标志蛋白,7个蛋白在肝细胞癌组织中表达上调.9个蛋白在肝细胞癌组织中表达下调,其中4.7 kDa,7.2 kDa和9.8 kDa蛋白峰差异性最明显;中分化和高分化肝细胞癌之间存在11个标志蛋白;通过查询蛋白库ExPasy并进行筛选,初步确定特定分子质量所对应的蛋白.结论:采用SELDI-TOF-MS技术,证实在肝细胞癌组织中存在多种高度特异性低分子蛋白.     

Identification of a new marker of hepatocellular carcinoma by serum protein profiling of patients with chronic liver diseases

Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS) is a proteomic technique that enables the profiling of proteins present in any biological material studied. We used this approach to identify new biomarkers of hepatocellular carcinoma (HCC) in the sera of patients with cirrhosis. Sera from 82 patients with cirrhosis, either without (n = 38) or with (n = 44) HCC, were analyzed by SELDI-TOF MS, and the results of the two groups were compared. The most efficient protein peaks leading to discrimination of patients with HCC were selected (receiver operative characteristic curves). The highest-scoring peak combination was established in a first group of serum samples (multinomial regression) and was tested in an independent group. The protein corresponding to the highest discrimination was purified and characterized further. The intensity of 30 protein peaks significantly differed between cirrhotic patients with and without HCC. An algorithm including the six highest-scoring peaks allowed correct classification (presence or absence of HCC) of 92.5% of patients in the test sample set and 90% in the validation sample set. The highest discriminating peak (8900 Da) was purified further and was characterized as the C-terminal part of the V10 fragment of vitronectin. An in vitro study suggested that the increase of the 8900-Da fragment in the serum of patients with HCC may proceed from the cleavage of native vitronectin with metalloproteases, a family of enzymes whose activity is enhanced in HCC. In conclusion, global protein profiling is an efficient approach that enabled us to identify a catalytic fragment ofvitronectin as a new serum marker of HCC in patients with chronic liver diseases.     

Application of Surface-Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Coupled With an Artificial Neural Network Model for the Diagnosis of Hepatocellular Carcinoma

Background/Aims: There are no satisfactory biomarkers for hepatocellular carcinoma (HCC). The surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technique has been used to identify biomarkers for cancer. Methodology: Four hundred thirty five serum samples were tested by SELDI-TOF-MS matching on a gold chip. Samples were assigned to a training set and a testing set according to collection order. The training set was used to identify statistically significant peaks and to develop the artificial neural network (ANN) model for diagnosing HCC. The testing set was used in a blind test to validate the diagnostic efficiency of the ANN model. Results: A total of 75 proteins that differed between patients and controls were identified (p<0.05). Seven of these proteins (p<0.01; m/z at 4207Da, 6604Da, 7734Da, 8106Da, 8545Da, 8599Da, 8894Da) were chosen to develop the ANN model. The model was subjected to a blind test using the testing set for HCC diagnosis. Sensitivity and specificity were 84.00% and 81.25%, respectively, and the accuracy was 81.90%. Conclusions: These results suggest that patients with HCC may have serum proteins that differ from healthy controls. The ANN is a new method for diagnosing and identifying HCC.     

HB virus infection and delta surinfection in Sahelian Africa

78 hospitalized patients were selected when presenting with at least one of these signs: hepatomegaly, jaundice, ascites, oesophageal varices, abdominal venous pattern, splenomegaly. All had radioimmunoassays for hepatitis B surface antigen (HBsAg) and antidelta antibody (78/78). Acute or chronic hepatic disease was diagnosed in 56 patients: 7 acute viral hepatitis, 13 chronic hepatitis, 23 non alcoholic hepatic cirrhosis, and 13 hepatocellular carcinoma. Twenty-two patients with other diagnoses served as controls. Serum antidelta was present in each group: acute viral hepatitis (2/7), chronic hepatitis (2/13), non alcoholic hepatic cirrhosis (9/23), hepatocellular carcinoma (3/13), controls (2/22). Every patient with acute or chronic hepatic disease and positive serum anti-delta was positive for serum HBsAg. Amony controls, 2 patients with positive serum antidelta were negative for serum HBsAg but positive for antiHBs. Delta superinfection is present in the sahelian region; Patients with acute viral hepatitis, chronic hepatitis, non alcoholic hepatic cirrhosis, and hepatocellular carcinoma are electively infected. Patients with acute or chronic hepatitis and positive serum antidelta have hepatitis B virus evolutive infection (positive serum HBsAg).