生物信息技术分析脑胶质瘤血清蛋白指纹图诊断模型的临床意义

目的探讨筛选的脑胶质瘤血清蛋白指纹图诊断模型的临床应用价值。方法用疏水性表面芯片(H4)和表面增强激光解吸离子化飞行时间质谱技术(SELDI-TOF—MS)及生物信息学分析方法与SPSS10．0软件，检测分析28例胶质瘤、37例其他脑良性肿瘤和40名健康人的血清蛋白指纹图，并建立脑胶质瘤血清蛋白指纹图诊断模型。结果用建立的区分脑胶质瘤与健康人的血清蛋白指纹图诊断模型进行盲法检测的准确率、敏感性和特异性分别为95．7％(22／23)、88．9％(8／9)和100％(14／14)。建立的区分胶质瘤与其他脑良性肿瘤的血清蛋白指纹图诊断模型，盲法检测脑胶质瘤的准确率、敏感性和特异性分别为86．4％(19／22)、88．9％(8／9)和84．6％(11／13)。建立的区分Ⅰ-Ⅱ级与Ⅲ-Ⅳ级脑胶质瘤的血清蛋白指纹图诊断模型的准确率分别为85．7％(13／15)、84．6％(11／13)。结论用SELDI—TOF—MS技术与生物信息分析法建立的3个血清蛋白指纹图诊断模型对脑胶质瘤的定性诊断提供了一条新途径。     

肺癌血清蛋白质指纹图谱诊断模型的建立

目的建立肺癌血清蛋白质指纹图谱诊断模型,探讨其对肺癌的诊断价值。方法收集127例肺癌患者、108例正常人血清标本,随机分成训练组(肺癌89例、正常人68例)与测试组(肺癌38例、正常人40例),应用表面增强激光解吸/电离-飞行时间-质谱(SELDI-TOF-MS)检测其蛋白质指纹图谱。用BioMarker Pattern软件分析训练组中肺癌与正常人血清中的差异蛋白后,建立肺癌诊断的最优分类树模型,然后用测试组血清标本盲法验证该模型的诊断效率。结果分类树诊断模型由质荷比分别为5 808、5 971和7 780的3种蛋白质组成。盲法验证显示,该模型诊断肺癌的准确率为92.3%,敏感性与特异性分别为92.1%(35/38)和92.5%(37/40)。结论肺癌血清蛋白质指纹图谱诊断模型较以往的传统方法具有更高的敏感性与特异性,值得进一步研究。     

应用SELDI-TOF-MS技术诊断2型糖尿病肾病

目的比较2型糖尿病(T2DM)肾病和对照人群血清蛋白质指纹图谱的差异,建立T2DM肾病诊断模型,探讨此技术对该病诊断的价值。方法采用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术检测51例T2DM肾病患者和66例对照人群血清,获得蛋白质指纹图谱。结合人工神经网络软件建立诊断模型并进行验证。结果在相对分子量2 000~30 000范围内共检测到175个蛋白峰,其中有17个蛋白峰明显表达差异(P<0.01)。筛选其中质荷比(m/z)分别为5 420、5 782、6 472、6 666、10 277和11 770的6个蛋白峰作为标志蛋白建立人工神经网络诊断模型。利用该模型对T2DM肾病进行盲法预测,结果表明其对该病的诊断敏感性和特异性分别为81.0%和96.2%。结论利用SELDI-TOF-MS和生物信息学技术建立了敏感性和特异性均较高的T2DM肾病诊断模型,为该病诊断提供了新途径。     

肾癌血清蛋白质指纹图谱诊断模型的建立

目的:筛选肾癌特异相关蛋白质,建立用于肾癌诊断的分类树模型,为进一步临床应用奠定基础。方法:采集肾癌患者48例与正常人83例血清,用表面增强激光解吸/电离飞行时间质谱分别检测其蛋白表达谱,用BioMarker Wizard软件筛选出差异蛋白,再用BioMarker Patterns软件建立肾癌诊断最优分类树模型。结果:在48例肾癌患者、83例正常人血清中共检测出44个差异蛋白质峰,在质荷比从2 800到16 100的差异蛋白中有21个蛋白质相对含量差异有统计学意义(P<0.05),从中选出12个差异蛋白建立分类树模型,用于鉴别肾癌患者与正常人,该模型在学习模式下的诊断准确率、灵敏度和特异性分别为98.47%(129/131),97.91%(47/48),98.79%(82/83),在测试模式下这3项指标分别为87.02%(114/131),85.42%(41/48),87.95%(73/83)。结论:肾癌血清蛋白质指纹图谱诊断模型具有一定优越性,为肾癌早期诊断提供了新途径。     

蛋白质指纹图谱技术在诊断白血病四种分型中的应用

目的对白血病典型的4种分型血清样本进行检测分析,建立白血病分型的蛋白质指纹图谱诊断模型。方法应用蛋白质指纹图谱技术检测16例急性粒细胞白血病M2a、18例慢性粒细胞白血病、14例急性淋巴细胞白血病及16例急性粒细胞白血病M3,采用Biomarker Wizard及Biomarker Patterns 5.0软件进行标记物的比较判别,建立决策树诊断模型。结果由m/z 3 376、8 055及9 421建立决策树诊断模型,其中16例急性粒细胞白血病M2a检出率为75.0%(12/16)、9例慢性粒细胞白血病检出率为100.0%(18/18)、14例急性淋巴细胞白血病检出率71.4%(10/14)、16例急性粒细胞白血病M3检出率为87.5%(12/14)。结论由3个蛋白标记峰构成的蛋白质指纹图谱诊断模型为白血病4种分型的诊断提供了新的借鉴和参考。     

DNA fingerprinting

Hypervariable tandem-repetitive minisatellite regions of human DNA can be used to generate individual-specific DNA fingerprints. Validation studies have demonstrated the reliability of the analysis, the mode of inheritance of the minisatellites, and the unparalleled degree of individual specificity. The uses of hypervariable probes in forensic biology, paternity testing, and the resolution of a wide range of problems in genetics, molecular biology, population biology, and medicine are illustrated.     

利用SELDI-TOF-MS蛋白质指纹图谱技术建立肺腺癌诊断模型

目的:寻找肺腺癌患者血浆(清)蛋白特异性生物标志物,用最佳的标志蛋白组合建立肺腺癌诊断模型,并探讨其用于肺腺癌筛查的可行性.方法:应用表面增强激光解吸电离飞行时间质谱(SELDI-TOF-MS)技术及CM10芯片,检测45例肺腺癌患者和40例正常人血清蛋白质指纹图谱,用Biomark Wizrd和Biomark Patterns Software软件对结果进行分析,建立肺腺癌诊断模型:双盲法分析另外19例肺腺癌和20例正常人血清的SELDI质谱数据,验证该模型诊断肺腺癌的准确度.结果:肺腺癌患者和正常人血清蛋白质指纹图谱间有31个差异蛋白(P<0.05),其中以分子量为11.66 kD的蛋白在两组血清中的差异最为显著(P<10-7),该蛋白对诊断肺腺癌有很强的敏感性和特异性;软件自动筛选出分子量为11.66 kD的标志蛋白为主根结点,联合3882.9、4673.2、3158.2和2047.2等4个标志蛋白建立了最佳的肺腺癌诊断模型,对肺腺癌诊断(盲法)的敏感性为84.2%(16/19),特异性为90.0%(18/20).结论:SELDI-TOF-MS技术建立的肺腺癌血清检测法,快速、简单、特异和敏感,为肺腺癌诊断和筛查提供了一个可行的途径和方法.     

血清蛋白质指纹图谱用于胃癌早期诊断的研究

目的:探讨血清蛋白质指纹图谱用于胃癌早期诊断的可行性。方法:用弱阳离子交换芯片蛋白芯片结合表面增强激光解吸电离飞行时间质谱技术检测血清蛋白质指纹图谱。以30例胃癌(10例早期、14例进展期、6例伴远隔脏器转移)与30例正常对照为训练组建立早期胃癌决策树分类器模型,并用盲筛方法对未参加建模的27例胃癌患者及7例良性病、19例健康者进行验证。结果:初步建立了3个对早期胃癌诊断预警分类效果较好的模型,参加盲筛的9例早期胃癌患者均被准确鉴别,模型的总体敏感性96.3%,特异性73.1%～84.6%;阴性预测值94.7%～95.7%;阳性预测值78.9%～86.7%。结论:虽然本实验建立的用于胃癌患者早期诊断预警模型在与胃良性病鉴别上效果不理想,但可有效用于人群中的早期胃癌初步筛查,有进一步研究的价值。     

Serum C-reactive protein in normal and infected neonates

Serum C-reactive protein determinations were performed on well and sick neonates, in order to gain information about normal values and its value in the diagnosis of neonatal septicaemia. The median value in 48 cord sera was 200 micrograms/l (range 15 to greater than 6,000 micrograms/L); there was no correlation between paired maternal and cord serum CRP levels (12 pairs). Thirty-six children were followed from birth for a mean of 20 days. There were 21 episodes of confirmed infection in 16 children, each associated with a sustained rise in C-reactive protein, often commencing before there was clinical evidence of infection. In four patients with raised levels, infection was suspected, but no firm evidence was obtained. In the remaining 16 children there were no values above 10 mg/l, and the 95th centile was about 6 mg/l, with no difference between values obtained in the first three days of life and those found subsequently. Hyaline membrane disease and jaundice were not associated with a rise. Raised serum C-reactive proteins is a good indicator of the presence of infection in the neonatal period.     

Serum eosinophil cationic protein in asthma of farmers

Objective: There is need for relevant markers of bronchial inflammation in epidemiologic studies of asthma. Serum eosinophil cationic protein (ECP) is a marker of eosinophil inflammation and asthma activity. We have studied serum ECP in atopic farmers with current asthma, in non-atopic asthmatics and in non-asthmatic, non-atopic controls. Methods and results: In a cross-sectional study of a representative sample of 8482 farmers in Norway, asthma was recorded using a self-administered questionnaire; spirometry and serum sampling were performed on all of them. Atopy was screened with Phadiatop and RAST analyses to the mites Lepidoglyphus destructor and Tyrophagus putrescentiae in all asthma cases and controls. All the identified atopics had additional RAST analyses on a set of allergens. Serum ECP was tested in 60 persons with current asthma and atopy (mean 16.2, 95% CI 13.2-19.3), 127 non-atopic asthmatics (mean 9.1, 95% CI 8.0-10.2) and 39 non-atopic controls (mean 5.5, 95% CI 4.0-7.0). ECP levels in atopic asthmatics were associated with number of positive allergens and reduction of FEV1 values. Moreover, the ECP levels were elevated with allergy to swine, cow, D. pteronyssinus, L. destructor, A. siro, T. putrescentiae, timothy grass and the cereal grains: wheat, oat, barley and rye. Conclusion: Serum ECP seems feasible as an indicator of inflammatory activity in epidemiological studies of current allergic asthma, and may help to indicate the importance of specific allergens. Although the ECP values were significantly more elevated in atopic than in non-atopic asthma, elevated serum ECP was not specific for atopic asthma.     

DNA fingerprinting in leukemia

Summary The detection of polymorphic regions of the human DNA provides individual specific markers, DNA fingerprints (DNA-F), for the identification of individuals. DNA-F can also be used for the detection of cell clones and serve as molecular markers of malignant disease. In leukemia DNA-F analysis can be used for the follow-up studies during remission or after bone marrow transplantation. Recent studies on DNA fingerprinting in leukemia are reviewed.     

Serum protein levels in critically Ill surgical patients

Serum protein levels were found to be markedly reduced in 44 critically ill surgical patients after they had been resuscitated in an intensive therapy unit (ITU). There was no difference in initial protein levels between those patients who survived and those who died, or between those patients who reacted to skin testing with recall antigens and those who showed no reaction. However, albumin and transferrin levels were lower in those patients who had major sepsis (p<0.02). Twenty-six of the patients remained in the ITU for more than five days. Repeat measurements made in these patients shortly before death or discharge from the ITU showed that prealbumin and transferrin levels were still not different in 17 survivors and 9 non-survivors. However, the mean albumin level was lower in the non-survivors (p<0.02), although nearly all of the survivors also continued to have low levels of albumin. There was no improvement in serum protein levels in 10 patients who, on skin testing, converted from being non-reactors to reactors.Although reduced serum protein levels were a common finding in critically ill patient, they were not clearly related to outcome or the response to recall skin antigens and this suggests that these tests may be altered by different mechanisms.