表面增强激光解析电离飞行时间质谱法筛选肺癌患者血清和肺泡灌洗液中标志蛋白的研究

Objective To detect the protein markers in serum and bronchoalveolar lavage fluid (BALF) of the patients with lung cancer by surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) technology, and to explore if they can be used as markers for the diagnosis of lung cancer.Methods SELDI-TOF-MS technology and protein chips weak cation exchange (WCX-2 chip) were used to detect the protein mass spectrum in serum and BALF of 35 patients with lung cancer and 18 cases of benign pulmonary diseases.The different protein markers were analyzed by Biomarker Pattern Software and the initial diagnosis models were set up.The diagnosis models were verified further by blind screen to confirm the efficacy of diagnosis.Results Five protein peaks in the sera of the patients with lung cancer were significantly higher (P ＜ 0.05 ).The protein peak with a mass/charge ratio (M/Z)of 5639 was selected to establish the classification tree model.The sensitivity of diagnosis was 80% (28/35) and the specificity was 78% (14/18).The results verified by blind screen showed a sensitivity of 85% (17/20),a specificity of 90% (9/10), a crude accuracy (CA) of 87% ( 26/30 ) and Youden' s index (γ) of 0.7.Eight protein peaks in the BALF of the patients with lung cancer were significantly higher ( P ＜ 0.05).The different protein peaks with M/Z of 7976 and 11 809 respectively were selected to establish the classification tree model.The sensitivity of diagnosis was 86% (30/35) and the specificity was 72% (13/18).The results verified by blind screen showed a sensitivity of 90% (18/20), a specificity of 90% (9/10), a CA of 90% (27/30) and γof 0.8.There was a complementary role in combination of differential proteins in serum and BALF and the sensitivity, specificity and accuracy of diagnosis for lung cancer were 100% by parallel test.Conclusions The SELDI-TOF-MS technology can screen out the differential protein markers in serum and BALF of the patients with lung cancer, which show high sensitivity and specificity as tumor markers.The differential proteins in the BALF may be more promising for clinical application.     

经纤维支气管镜生物性肺减容术的动物实验

目的:探讨生物性肺减容术糜蛋白酶代替胰蛋白酶的合适剂量、国产生物蛋白胶中纤维蛋白原与凝血酶的最佳配比、706代血浆替换小牛血清的可行性.方法:9头健康约克幼猪(体重16～22 kg)分别标记1～9号.第一阶段用不同剂量(A,B两种)糜蛋白酶和不同配比(A',B'两种)生物蛋白胶配伍加上小牛血清通过纤维支气管镜给药于1～...     

窒息性气管狭窄四例救治体会

因病变导致气管接近完全堵塞而处于极度缺氧状态,患者可能出现缺氧性心脏骤停并死亡,这种严重的气管狭窄临床上称为窒息性气管狭窄,需要紧急处理.能否快速调整氧合状态、防止缺氧性心脏骤停是抢救成败的关键,既往多采用外科手术治疗[1-2].现将4例典型的结核所致窒息性气管狭窄患者采用体外循环下球囊扩张及气管插管扩张的诊治经验报道如下.     

白细胞介素32在肺癌患者支气管肺泡灌洗液及血清的表达

目的研究肺癌患者支气管肺泡灌洗液（BALF）和外周血清中白细胞介素32（interleukin-32,IL-32）表达及其临床意义。方法采用酶联免疫吸附测定（ELISA）技术测定40例肺癌患者及21例良性肺病变患者BALF和血清IL-32的浓度。结果①在BALF中肺鳞癌组患者IL-32（156.49±32.84）ng/L显著低于肺腺癌组（224.70±54.00）ng/L及肺良性病变组（219.06±67.82）ng/L（P〈0.05）,而血清IL-32浓度无论是肺鳞癌组（149.08±31.31）ng/L还是肺腺癌组（158.03±46.58）ng/L均低于肺良性病变组（188.11±45.98）ng/L（P〈0.05）;②Ⅳ期肺癌患者BALF中IL-32（221.43±53.02）ng/L高于Ⅲ期（157.68±37.56）ng/L和Ⅱ期（135.06±21.03）ng/L（P〈0.05）,而血清IL-32浓度尽管在Ⅱ期（122.84±25.23）ng/L、Ⅲ期（147.59±32.52）ng/L、Ⅳ期（158.60±39.01）ng/L肺癌患者逐渐升高,但三者浓度差异无统计学意义（P〉0.05）。结论 IL-32在肺良性病变及肺腺癌患者BALF中的高表达水平提示其可能参与肺部感染性疾病和肺腺癌的发病机制,肺癌患者BALF中IL-32浓度与肺癌TNM分期有关,表明它有可能成为肺癌的病程进展及预后不良一个指标。     

应用SELDI-TOF-MS技术筛选肺鳞癌和肺腺癌患者差异蛋白的研究

目的 应用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术筛选出不同病理类型肺癌患者血清、支气管肺泡灌洗液(BALF)和肺癌组织中的差异蛋白,并探讨其临床意义.方法 选用WCX-2芯片、SELDI-TOF-MS技术检测20例肺鳞癌、20例肺腺癌和20例肺良性病变患者血清、BALF和肺组织匀浆的蛋白质谱,用Biomarker Pattern软件分析肺鳞癌和肺腺癌的差异蛋白并初步建立诊断模型.结果 ①肺鳞癌组与肺良性病变组:在血清、BALF、肺组织匀浆中发现存在2、9、8个差异蛋白峰(P＜0.05),分别选用其中质荷比为5 124.24、7 967.29联合10 843.45、7914.59联合8709.66的差异蛋白波峰建立分类树模型,其诊断肺鳞癌的灵敏度分别为85％、80％、75％,特异度分别为65％、80％、90％,正确率分别为75％、80％、83％,阳性预测值分别为71％、80％、88％,阴性预测值分别为81％、80％、78％,其相对应ROC曲线下面积分别为0.750、0.916、0.930.②肺腺癌组与肺良性病变组:在血清、BALF、肺组织匀浆中发现存在8、9、7个差异蛋白峰(P＜0.05),分别选用其中质荷比为9295.79、7923.01、2452.49的差异蛋白波峰建立分类树模型,其诊断肺腺癌的灵敏度分别为75％、80％、70％,特异度分别为65％、90％、85％,正确率分别为70％、85％、78％,阳性预测值分别为68％、89％,82％,阴性预测值分别为72％、82％、71％,其相对应ROC曲线下面积分别为0.844、0.933、0.825.结论 肺鳞癌和肺腺癌BALF及肺组织匀浆中的差异蛋白较血清多,诊断效率比血清高,肺鳞癌BALF中质荷比为7967.29和10843.45的差异蛋白峰联合、肺组织匀浆中质荷比为7914.59和8709.66的差异蛋白峰联合建模及肺腺癌BALF中质荷比为7923.01的差异蛋白峰建立分类树诊断模型,其诊断的灵敏度、特异度和正确率可达到75％～90％,有可能作为诊断肺鳞癌和肺腺癌的标志蛋白.     

表面增强激光解析电离飞行时间质谱技术对周围型肺癌差异蛋白表达的研究

目的 筛选周围型肺癌患者血清、肺组织活检前后BALF中的差异蛋白,探讨其表达特点、影响因素及其临床意义.方法 选择2008年3月至2009年11月周围型肺癌及肺部良性病变患者各20例,应用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术分别检测患者血清、活检前后BALF中的蛋白质质谱并初步建立分类树诊断模型.结果 (1)两组患者血清中检出6个差异蛋白峰,以质荷比为6637的差异蛋白建模诊断周围型肺癌的敏感度为70%(14/20),特异度为90%(18/20),正确率为80%(32/40),阳性预测值为88%(14/16),阴性预测值为75%(18/24),受试者工作曲线(ROC曲线)下而积(AUC)为0.73;(2)两组活检前BALF中检出11个差异蛋白峰,以质荷比为7982的差异蛋白建模诊断周围型肺癌的敏感度为85%(17/20),特异度为90%(18/20),正确率为88%(35/40),阳性预测值为89%(17/19),阴性预测值为86%(18/21),AUC为0.94;(3)两组活检后BALF中检出14个差异蛋白峰,以质荷比为7671的差异蛋白建模诊断周围型肺癌的敏感度为85%(17/20),特异度为100%(20/20),正确率为93%(37/40),阳性预测值为100%(17/17),阴性预测值为87%(20/23),AUC为0.93.结论 周围型肺癌活检前后BALF中检出的差异蛋白较血清中多,活检后BALF中的差异蛋白较活检前多.活检前质荷比为7982的差异蛋白与活检后质荷比为7671的差异蛋白在周围型肺癌患者BALF中的AUC值均＞0.9,高于血清;这2种差异蛋白可作为较好的早期诊断周围型肺癌的标志蛋白.

Abstract：

Objective To scan the protein mass spectra in the sera and bronchoalvolar lavage fluid (BALF) from patients with peripheral lung cancer, screen out the differential proteins, and explore the clinical significance of the differential proteins. Methods SELDI-TOF-MS was used to detect the protein mass spectra and to screen out the differential proteins in the sera and BALF collected before and after lung biopsy in 20 patients with peripheral lung cancer and 20 patients with benign pulmonary diseases. The differential proteins were analyzed and the initial diagnostic models were set up. Results ( 1 ) There were 6 differential protein peaks in the sera of the 2 groups ( P ＜0. 05 ＝. The protein with a mass/charge ratio ( M/Z) of 6637 was selected to establish the diagnostic model. The sensitivity of diagnosing peripheral lung cancer was 70% (14/20) ,the specificity 90% ( 18/20), the accuracy 80% (32/40), the positive predictive value ( PV + ) 88% ( 14/16), the negative predictive value( PV - ) 75% ( 18/24), and the area under the ROC curve (AUC)was 0.73. (2) There were 11 differential protein peaks in the BALF collected before lung cancer biopsy of the 2 groups ( P ＜ 0. 05 ＝. The protein with a M/Z of 7982 was selected to establish the diagnostic model. The sensitivity of diagnosing peripheral lung cancer was 85% ( 17/20 ), the specificity 90% ( 18/20), the accuracy 88% (35/40), the PV + 89% ( 17/19), the PV - 86% ( 18/21 ), and the AUC was 0. 94. (3) There were 14 differential protein peaks in the BALF collected after lung cancer biopsy of the 2 groups ( P ＜0. 05 ＝. The protein with a M/Z of 7671 was selected to establish the diagnostic model.The sensitivity of diagnosing peripheral lung cancer was 85% (17/20) ,the specificity 100% (20/20), the accuracy 93% (37/40), the PV + 100% (17/17), the PV- 87% (20/23), and the AUC was 0. 93. Conclusions There were more differential proteins in BALF as compared with sera. There were more differential proteins in the BALF collected after lung biopsy as compared to that before lung biopsy. The AUC of the diagnostic models set up by proteins in BALF collected before and after lung biopsy were all above 0. 9 and showed higher efficiency for the diagnosis of peripheral lung cancer as compared to proteins in sera.These differential proteins may be better tumor markers for the diagnosis of peripheral lung cancer at the early stage.     

难治性恶性气管-支气管狭窄的治疗体会

目的 探讨难治性恶性气管-支气管狭窄的有效处理方法.方法 回顾性分析2006～2010年间在该院住院的3例肺癌术后复发致气管-支气管狭窄、3例甲状腺癌气管转移致气管狭窄、2例结肠癌术后复发致气管狭窄以及1例巨大纵隔原发精原细胞癌致气管狭窄患者的诊疗过程.结果 1例肺癌复发患者绕过气管插管用纤支镜定位法将1镍钛金属支架置入于气管-左主支气管,其余2例肺癌复发者在使用高频电刀切除瘤体后将金属支架放置在气管-支气管;3例甲状腺癌患者在拔除气管插管后,1例置入2枚金属支架、1例置入1枚气管支架而迅速解除呼吸困难症状,1例置入2枚气管支架都不能缓解其呼吸困难;精原细胞癌患者经气管插管置入1枚镍钛金属支架后顺利撤机和拔除气管插管.结论 肺癌复发引起的气管狭窄,可对气管内肿瘤组织用高频电刀予以切除,然后置入气管支架;转移性肿瘤致外压性气管狭窄者可绕过或经气管插管采用纤支镜定位法置入镍钛金属支架.     

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。