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.     

应用蛋白质组学技术对脂肪肝患者血清标志物的筛选

目的:用纳米磁珠与基质辅助激光解析离子化飞行时间质谱(MALDI-TOF-MS)技术检测脂肪肝患者血清蛋白质组,筛选候选标志物并建立诊断模型,初步探讨所建立的诊断模型在脂肪肝早期诊断中的临床意义.方法:95例中度和76例轻度脂肪肝患者均经B超证实,99名健康人血清.用WCX(弱阳离子交换型)纳米磁珠检测各血清标本获得血清蛋白指纹图谱.用Biomarker Wizard软件分析所获得的蛋白图谱找出差异蛋白,再用Biomarker Patterns建立诊断模型.扩大样本量,通过盲法分析进一步验证诊断模型的可靠性.结果:初始建模时,建立了由m/z为7626.2、24147.7、6118.2这3个差异蛋白峰组成的中度脂肪肝诊断模型,其敏感性为90.2%,特异性为93.9%.扩大样本盲法验证的敏感性为90.9%,特异性为92.0%.另选取了m/z为7626.2、3286.0、2760.7、2543.1、2746.1这5个差异蛋白峰组成的模型,能把轻度从中度脂肪肝患者中区分出来:选取m/z为7626.2、11721.0、24141.7、8529.1、9042.7这5个差异蛋白峰组成的模型,能将轻度脂肪肝患者从健康对照中区分出来.结论:蛋白质组学技术可筛选出有意义的血清生物标志差异蛋白.由三个差异表达蛋白及其特定组合构成的诊断模型可以区分中度脂肪肝患者与健康人,由五个特定组合构成的差异表达蛋白的模型能区分轻度、中度脂肪肝患者及区分轻度脂肪肝患者与健康对照.     

原发性胆汁性肝硬化血清蛋白质谱的初步研究

目的筛选与原发性胆汁性肝硬化（PBC）相关的血清蛋白质分子标志物并建立指纹诊断模型。方法收集30例PBC患者血清,30例年龄、性别相匹配正常人,采用弱阳离子磁珠（MB-WCX）为检测介质,基质辅助激光解吸电离质谱（MADIL-TOF-MS）检测得到蛋白质谱,再应用基于磁珠的多维纳升级高效液相色谱及液体蛋白芯片指纹图谱（CLINPROT）系统比较两组血清蛋白质谱。结果在m/z100～10000范围内,检测出的129个蛋白峰中11个蛋白峰差异有统计学意义（P〈0.05）,其中m/z为4963.6、5805.33、1544.45、2292.16、2466.45、2845.85在PBC中表达上调,而m/z为3262.24、3191.2、4090.5、4072.43、3277.46的蛋白峰在PBC中下调;选择m/z为3262.24、3191.2这2个蛋白峰建立的蛋白芯片指纹图谱（CLINPROT）系统对PBC诊断的敏感性为90%,特异性为95%。结论筛选出的11个蛋白分子标志物可能与PBC有关,其中m/z4963在PBC中表达最明显;蛋白芯片指纹图谱（CLINPROT）系统可能对PBC的诊断具有一定临床意义。     

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.     

胃癌与慢性胃炎的唾液蛋白指纹图谱鉴别诊断模型

目的:筛选胃癌患者与慢性胃炎患者及健康志愿者的唾液蛋白质组差异表达谱,并建立胃癌患者与慢性胃炎唾液无创伤分子诊断(判别)模型.方法:采集57例胃癌患者(术前且无放化疗)和28例慢性胃炎患者的唾液标本,用WCX(弱阳离子交换型)纳米磁珠联合基质辅助激光解析离子化飞行时间质谱(matrix assisted laser desorption ionization time-offlight mass spectrometry,MALDI-TOF-MS)技术进行检测,获得各标本的蛋白指纹图谱.用Biomarker Wizard软件分析所获得的蛋白指纹图谱找出差异蛋白,再用Biomarker Patterns 5.0.2建立鉴别诊断模型.结果:优化筛选建立了由m/z为4267.09、6564.85、2138.14等3个差异蛋白峰组成的胃癌与慢性胃炎的鉴别诊断模型,经临床回代检验该分子诊断模型对鉴别胃癌与慢性胃炎的灵敏度和特异度分别为96%(55/57)和86%(24/28),通过交叉验证法进一步验证诊断模型的可靠性,结果该模型的灵敏度和特异度分别为89%(51/57)和75%(21/28).结论:基于WCX结合MALDI-TOF-MS技术建立的唾液蛋白组分子诊断模型为胃癌与慢性胃炎的鉴别诊断提供了一种敏感无创的新方法.     

胰腺癌血清蛋白质波谱模型的构建及诊断价值研究

目的 建立胰腺癌诊断模型,寻找与胰腺癌分期相关的蛋白峰并鉴定出新型的肿瘤标志物.方法 应用表面增强激光解吸离子化飞行时间质谱(SELDI-TOF-MS)技术,用SAX2蛋白芯片,检测胰腺癌患者、胰腺良性疾病患者和正常对照者的血清蛋白指纹谱,统计建立决策树诊断模型和Logistic回归模型并评估诊断价值,蛋白芯片免疫法鉴定差异蛋白峰,ELISA法测定其在血清中的浓度.结果 胰腺癌组和正常对照组指纹谱比较.26个蛋白峰差异有统计学意义(P＜0.01),胰腺癌组和胰腺良性疾病组指纹谱比较,16个蛋白峰差异有统计学意义(P＜0.05).建立的决策树诊断模型对胰腺癌的敏感性为83.3%,特异性为100.0%,经ROC曲线评估,该模型优于糖链抗原(CA)19-9.有6个差异蛋白峰在不同分期咦腺癌中差异有统计学意义(P＜0.01),建立Logistic回归模型诊断早期胰腺癌.敏感性为81.6%,特异性为80.6%.鉴定出差异蛋白峰质荷比(M/Z)28068为C140H166,其诊断胰腺癌的敏感性＞82%,特异性＞88%.结论 应用SELDI-TOF-MS技术建立的诊断模型对胰腺癌的诊断有较大价值,明显优于CA19-9.鉴定出的差异蛋白C140rf166有望在胰腺癌诊断中发挥作用.     

应用蛋白质指纹技术筛选强直性脊柱炎血清特异性标志物的临床研究

目的应用表面增强激光解析离子化-飞行时间质谱技术(surface-enhanced laser desorption ioni zation/time of flight mass spectrometry,SELDI-TOF-MS)和蛋白质芯片筛选强直性脊柱炎(ankylosing spondylitis,AS)患者血清特异性标志物,用于疾病诊断、评估及预测病情进展。方法采用SELDI-TOF-MS技术和弱阳离子交换(weak cation exchange)芯片检测2008年4月至2009年1月山西医科大学第二医院风湿免疫科收治的69例AS患者及12名健康对照者、10例类风湿关节炎(rheumatoid arthritis,RA)患者血清蛋白质表达,进一步将AS患者分为活动期与非活动期,中轴关节受累及中轴、外周关节均受累,HLA-B27阳性与阴性组,比较不同分组之间患者血清蛋白质指纹图谱,采用SELDI质谱仪自带的Biomarker Wizard和Biomarker Pattern软件筛选,初步建立疾病诊断预测模型。结果由8085、2640和2932建立的诊断预测模型Ⅰ诊断AS的敏感度为94.23%,特异度为100%。由3677、3880、2539、3159和3242建立的诊断预测模型Ⅱ判断病情活动的敏感度为98.11%,特异度为100%。由4700、8687和18538建立的诊断预测模型Ⅲ预测AS同时有中轴及外周关节受累的敏感度为80.00%,特异度为82.35%。由10259、7972、2048、2154和2954建立的诊断模型Ⅳ区分AS和RA的敏感度为100%(69/69),特异度为100%(10/10)。结论通过SELDI-TOF-MS技术建立的血清蛋白质指纹图谱可以筛选AS患者血清中的特异性蛋白质标志物,有望成为诊断疾病及评估病情的一种初筛平台。     

应用SELDI-TOF-MS技术建立肝癌筛选血清蛋白质指纹图谱模型

目的:建立肝癌筛选血清蛋白质指纹图谱模型．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蛋白芯片获得新发肝癌、肝硬化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,建立肝癌的筛选模型．结果:肝癌患者与健康对照组血清蛋白质指纹图谱之间有5个标志蛋白(4477,8943,5181, 8617,13 761 Da)在肝癌患者血清中高表达,肝癌患者与肝硬化患者血清蛋白质指纹图谱之间2个标志蛋白(4477,13 761 Da)在肝癌患者血清中高表达,1个标志蛋白(4097 Da)在肝癌患者血清中低表达．SELDI-TOF-MS技术的特异性(60／60,100％);敏感度(18／20,90％)．分析系统筛选出4477,8943,13 761,4097 Da标志蛋白建立的肝癌诊断模型．结论:建立的血清蛋白质指纹图谱模型能够区分肝癌与非肝癌患者,SELDI-TOF-MS在肝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．     

应用蛋白质指纹图谱技术快速鉴别诊断菌阴肺结核

目的 探索应用蛋白质指纹图谱技术于菌阴肺结核与肺炎的鉴别诊断。方法 从本院临床病例中,选择菌阴肺结核和肺炎患者及健康者各60例,应用表面加强激光解吸电离飞行时间质谱技术(SELDI/ToF-Ms)和蛋白芯片技术检测血清蛋白,并应用Ciphergen蛋白芯片3.1.1软件进行比较,分析其相关蛋白峰值并进行统计学处理。结果 对180例菌阴肺结核、肺炎患者、健康者的血清蛋白指纹图谱数据进行比较,发现有5个蛋白峰(1 028.49、4 796.56、7 564.77、8 048.02、11 526.75 m/z)存在显著的差异,有统计学意义(P<0.01)。由这5个蛋白峰组成的诊断模型鉴别诊断菌阴肺结核与肺炎的总有效率84.2%(101/120),敏感性与特异性分别为82.5%(52/63),85.9%(49/57);阳性预测值86.7%(52/60),阴性预测值为81.7%(49/60)。诊断模型在判别肺炎、菌阴肺结核患者与健康者之间,总有效率达89.4%(161/180),特异性为100%(60/60),灵敏度为84.2%(101/120),阳性预测值100%(101/101),阴性预测值75.9%(60/79)。结论 蛋白质指纹图谱技术具有方法简便、检测快速,标本用量少的优点,是筛选结核病特异性标志物的有效手段,通过蛋白质指纹图谱技术检测,发现了具有良好鉴别诊断的“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.     

Options for screening for colorectal cancer

Unlike other types of cancer, there are several options for screening for colorectal cancer (CRC). The most extensively examined method, faecal occult blood testing (FOBT), has been shown, in three large randomized trials, to reduce mortality from CRC by up to 20% if offered biennally and possibly more if offered every year. Recently published data from the US trial suggest that CRC incidence rates are also reduced by up to 20%, but only after 18 years. In this study, the number of positive slides was associated with the positive predictive value both for CRC and adenomas larger than 1 cm, suggesting that the reduction in CRC incidence was caused by the identification and removal of large adenomas. In this respect, this study supports the concept that removing adenomas prevents CRC. More efficient methods of detecting adenomas include the use of colonoscopy or flexible sigmoidoscopy (FS). Considerable evidence exists from case-control and uncontrolled cohort studies to suggest that endoscopic screening by sigmoidoscopy reduces incidence of distal colorectal cancer. However, in the absence of evidence from a randomized trial, several countries have been reluctant to introduce endoscopic screening. Three trialsare currently in progress (in the UK, Italy and the US) to address this issue. Two of these trials are examining the hypothesis that a single FS screen at around age 55-64 might be a cost-effective and acceptable method for reducing CRC incidence rates. Recruitment and screening are now complete in both studies and the first analysis of results on incidence rates is expected in 2004. Colonoscopy screening at 10-year intervals has recently been endorsed in the US on the basis that the reductions in incidence observed with distal CRC screening can be extrapolated to the proximal colon. However, data are lacking and a pilot study for a trial of the acceptability and efficacy of colonoscopy screening is in progress in the US. It has also been suggested that FOBT testing should be used to detect proximal CRC missed by sigmoidoscopy screening, but the small amount of published data suggest that supplementing FS with FOBT offers very little advantage over FS alone. Other forms of CRC screening are under investigation and represent exciting options for the future. Extraction of DNA from stool is now feasible and a number of research groups have shown high sensitivity for CRC using a panel of DNA markers including mutations in k-ras, APC, p53 and BAT26. Data so far indicate that, with the exception of k-ras, these markers are highly specific and therefore represent a significant improvement over FOBT. Whether these tests will replace or supplement existing methods of screening has yet to be determined. It has been suggested that BAT26, which is a marker of microsatellite instability, a feature of proximal sporadic CRC, might be a useful adjunct to sigmoidoscopy screening. Others have suggested that a test for occult blood should be included with the DNA markers to further increase sensitivity. It is not yet known how sensitive these markers are for adenomas--it is only by detecting adenomas that CRC incidence rates can be reduced. A final exciting new option for screening is virtual colonoscopy (VC), which by screening out people without neoplasia allows colonoscopy to be reserved for patients requiring a therapeutic intervention. The sensitivity of VC for large adenomas and CRC appears to be high, although results vary by centre and there is a steep learning curve. Sensitivity for small adenomas is low, but perhaps it is less essential to find such lesions. Some groups have suggested that virtual colonoscopy might be a useful option for investigating patients who test positive with stool-based screening tests. Whichever CRC screening method is finally chosen (and there is no reason why several methods should not ultimately be available), high quality endoscopy resources will always be required to investigate and treat neoplastic lesions detected.     

Indications for surgery for aortic regurgitation

Aortic valve replacement for isolated aortic regurgitation (AR) is usually not indicated unless the regurgitation is severe. However, not all patients with severe AR require aortic valve replacement. This review focuses on the causes of AR and the pathophysiology of acute versus chronic AR, and the attendant adaptive mechanisms of the left ventricle that ultimately determine their different natural histories. Aortic valve surgery must be performed in a timely manner to prevent cardiac death, ameliorate symptoms, and limit late postoperative excess mortality.