肺癌脱落细胞端粒酶活性检测的临床意义

目的：探讨肺癌脱落细胞端粒酶活性检测的临床意义。方法：收集63例肺癌患者和31例非肺癌肺疾病患者的支气管肺泡灌洗液，同时行刷片和灌洗液细胞学检查；34例肺癌患者痰液，31例非肺癌肺疾病患者痰液；彩PCR－TRAP银染法检测端粒酶活性。结果：肺癌和非肺癌肺疾病支气管肺泡灌洗液端粒酶活性阳性率分别为76．2％（48／63）和6．5％（2／31），P＜0．01；肺癌支气管肺泡灌洗液端粒酶活性阳性率高于刷片细胞学阳性率（58．7％，37／63），P＞0．05；高于灌洗液细胞学阳性率（14．7％，9／63），P＜0．01。肺癌痰液和非肺癌肺疾病痰液端粒酶活性阳性率分别为29．4％（10／34），3．2％（1／31），P＜0．01。结论：肺癌端粒酶活性检测有较高的特异性和敏感性，可应用于肺癌的临床诊断。支气管肺泡灌洗液和痰液端粒酶检测能提高肺癌检出率。     

癌性胸水细胞端粒酶活性研究

目的：研究癌性胸水细胞中的端粒酶活性。方法：采用PCR－TRAP方法检测癌性胸水细胞标本中的端粒酶活性,并将检测结果与细胞学诊断结果进行比较。结果：57例诊断明确的癌性胸水细胞标本经细胞学检查和端粒酶活性检测发现：在43例肺癌胸水标本中,29例细胞学阳性胸水有27例端粒酶阳性,3例细胞学可疑胸水有2例端粒酶阳性,11例细胞学阴性胸水有4例端粒酶阳性。在14例其它肿瘤胸水标本中,8例细胞学阳性及1例细胞学可疑胸水标本端粒酶检测均为阳性,5例细胞学阴性胸水有1例端粒酶阳性。细胞学诊断和端粒酶检测总阳性率分别为64．91％（37／57）和75．44％（43／57）。结论：结果提示端粒酶活性检测可能在癌性胸水诊断和鉴别诊断方面有重要辅助诊断价值。     

CT扫描与痰液肿瘤标志物检测对周围型肺癌的早期诊断价值

目的：研究CT扫描与肺活检后痰液脱落细胞的端粒酶活性和p16甲基化检测对早期周围型肺癌的诊断意义。方法：用PCR－TRAP－ELISA半定量及PCR－TRAP银染定性法、甲基化相关的PCR法，前瞻性检测55例经CT扫描而发现的肺部孤立性结节（直径≤30mm)、并疑诊为早期周围型肺癌的患者（T1N0M0），行CT导向肺活检（纤维支气管镜肺活检33例，经皮肺针刺活检22例）后24h内痰液脱落细胞端粒酶活性及p16甲基化状态，并将检测结果与组织病理进行对照研究。结果：CT扫描对周围型肺癌诊断的敏感性为100％，但特异性只有61．8％（34／55）。端粒酶活性的敏感性为79．4％，特异性为90．5％，准确性为83．6％；p16甲基化的敏感性为32．4％，特异性为100％，准确性为58．2％，3者联合检测的敏感性为86．1％，特异性为90．5％，准确性为87．3％。对照组端粒酶阳性率9．5％（2／21），无1例检测到p16甲基化。结论：CT扫描与端粒酶活性、p16甲基化联合可弥补痰液的细胞学检查的不足，提高肺癌痰检的敏感性，有助于周围型肺癌早期诊断和肺部孤立性结节的鉴别诊断。     

痰检、纤支镜下刷检联合检测提高肺癌诊断的阳性率

痰检、纤支镜下刷检联合检测提高肺癌诊断的阳性率湖北省荆沙市胸科医院陈从德诊断原发性支气管癌（肺癌）的方法很多，细胞学是重要的手段之一，其中痰液脱落细胞学检查（痰检）和纤维支气管镜（纤支镜）下毛刷涂片细胞学检查（刷检）各有其优点，但皆有假阴性。本文通过...     

纤支镜刷检、活检、冲洗、检后痰对非典型肺癌的诊断价值

纤支镜刷检、活检、冲洗、检后痰对非典型肺癌的诊断价值王琳主治医师曹培根解放军８５医院（２００２３３）据报道，肺癌病人的纤支镜检查（ＦＢ）阳性率，刷检８５％，活检９４％～９７％，总细胞学阳性率８５％，而纤支镜检查对胸部Ｘ线未显示明显增生性块影的非典型肺...     

痰和纤维支气管镜刷片细胞学检查在肺癌诊断中的意义

目的探讨痰和纤支镜刷片细胞学检查在肺癌诊断中的意义。方法收集532例同时进行痰和纤支镜检查的肺癌病例,分析痰和纤支镜刷片细胞学检查的敏感性和分型的准确性,与纤支镜咬检组织学对比,评价细胞学在肺癌诊断中的意义。结果痰细胞学的敏感性为33．8％,纤支镜刷片细胞学的敏感性为58．7％,纤支镜咬检组织学的敏感性为39．1％。痰细胞学分型诊断与组织学的符合率鳞癌为95．7％,腺癌为87．2％,小细胞癌为100．0％,痰细胞学区分小细胞癌和非小细胞癌的准确性为100．0％。纤支镜刷片细胞学分型诊断与组织学的符合率鳞癌为93．3％,腺癌为85．4％,小细胞癌为95．5％,纤支镜刷片细胞学区分小细胞和非小细胞癌的准确性为98．0％。结论纤支镜刷片细胞学具有较高的敏感性,在肺癌的诊断中有重要的应用价值。痰细胞学敏感性较低,但可作为纤支镜检查阴性和不能耐受纤支镜检查的肺癌患者的补充检查手段。两者的细胞学分型具有较高的可信度。     

胃癌及癌前病变组织中端粒酶RNA表达及其临床意义

目的 探讨胃癌及癌前病变胃粘膜端粒酶RNA与端粒酶活性检测及临床意义。方法 分别采用原位逆转录－PCR、端粒重复序列扩增法（TRAP）检测114例胃粘膜组织标本端粒酶RNA与端粒酶活性,其中包括慢性浅表性胃炎（CSG）32例、不典型增生（AH）34例、胃癌（GC）48例。结果 原位逆转录－PCR检测胃粘膜活检标本的端粒酶RNA阳性率为57．9％（66／114）,显著高于TRAP法检测端粒酶活性检出率（44．7％,51／114,P＜0．05）。在不同胃粘膜病变中,AH及GC组的端粒酶RNA阳性率分别为52．9％、100％,而CSG组胃粘膜中未检出端粒酶RNA;AH及GC组的胃粘膜端粒酶RNA阳性率分别显著高于CSG（P＜0．05）,而GC组端粒酶RNA阳性率亦明显高于AH组（P＜0．05）。端粒酶RNA主要分布于胃粘膜癌细胞及癌前病变上皮细胞的胞核内。结论 端粒酶RNA表达与胃癌的发生密切相关。原位逆转录－PCR检测胃粘膜端粒酶RNA可能是较端粒酶活性更灵敏的生物学指标,对胃粘膜癌变的预测和早期诊断有重要价值。     

影响纤支镜下刷检阳性率的因素

纤维支气管镜（纤支镜）下毛刷涂片细胞学检查（刷检）作为诊断肺癌的重要手段之一，有着其他检测不可替代的优点，但文献报道的阳性率差异甚大［１］。本文通过对１２０例肺癌病例纤支镜下刷检结果进行分析，探讨影响其阳性率的因素。材料与方法一、对象：选择１９８５年...     

肺癌支气管肺泡灌洗液标本端粒酶活性检测的价值

目的:探讨纤维支气管镜检查所取支气管肺泡灌洗液(BALF)标本端粒酶活性检测对肺癌诊断的价值.方法:应用TRAP-PCR-ELISA方法及TRAP-银染法分别检测60例支气管肺泡灌洗液标本端粒酶活性及活性水平并分析对肺癌诊断的价值.结果:41例肺癌组标本端粒酶活性及活性水平与癌疑组相近,但明显高于炎症对照组.端粒酶活性的检测敏感性83.8%,特异性69.6%,准确性80.4%.周围性肺癌端粒酶活性阳性率83.3%,虽较中央性肺癌组高,但两者比较无显著性差异.病理确诊的肺癌BALF端粒酶活性阳性率83.8%,明显高于刷检细胞学检查48.6%,两者联合平行试验敏感性91.7%,特异性60.5%.用TRAP-银染法测定端粒酶活性率与定量法相近,但端粒酶活性水平较低的标本定型分析为弱阳性,条带不清晰或条带较少.结论:端粒酶活性是肺癌的标志物之一,用定量法检测支气管肺泡灌洗液标本端粒酶活性与组织细胞学检查相结合,可提高肺癌的早期诊断率.     

A comparative study of telomerase activity in sputum, bronchial washing and biopsy specimens of lung cancer.

The potential of telomerase, the ribonucleoprotein enzyme, as a non-invasive screening marker was studied in pre-bronchoscopy sputum (S), bronchial washings (W) and bronchoscopic biopsy (B) samples from individuals under evaluation for lung cancer. Out of the 52 cases studied, 42 were clinically suspected primary lung cancer patients and 10 had pulmonary disorders but had no clinical evidence of lung cancer. Fifteen (39.5%) S samples, 24 (63.1%) W samples and 32 (84.2%) B samples, which were cytologically/histopathologically positive were also positive for telomerase activity. Interestingly, 16 (42%) S samples, 20 (52.6%) W samples and 20 (52.6%) B samples, initially reported cytologically/histopathologically negative, showed detectable telomerase activity. Lung cancer was finally confirmed in these cases by repeat cytology/histopathology. However, telomerase activity was detected in 31 (81.6%) S, 26 (68.4%) W and 33 (86.8%) B samples of suspected lung cancer patients. Telomerase activity was negative in S, W, and B of four of the suspected cases, which ultimately turned out to be negative for lung cancer. Cytopathology/histopathology alone (including repeat attempts) identified 15 (39.5%) cases of sputum, 24 (63.1%) cases of bronchial washings and 32 (84.2%) bronchoscopic biopsy samples. Out of 10 controls, low telomerase activity was detected in only one (10%) of the bronchial washings, which later turned out to be due to large number of inflammatory cells. Telomerase activity assay of sputum carried sensitivity, specificity and diagnostic accuracy of 81.6, 100 and 86.5%, respectively, while that for bronchial washing was 68.4, 100 and 76.9%, respectively, and for bronchoscopic biopsy samples was 86.8, 100 and 88.1%, respectively. A positive correlation (P<0.01) was seen between age and telomerase activity in sputum, bronchial washing and biopsy samples but no significant correlation was seen between sex and telomerase activity or duration of smoking and telomerase activity. A significant positive correlation was observed between staging and telomerase activity in sputum (P<0.01), bronchial washing (P<0.01) and biopsy samples (P<0.01). Our findings indicate that telomerase is a specific marker for malignant lung disease and can complement cytology/histopathology in the diagnosis of lung cancer. Sputum telomerase assay holds the potential for early and non-invasive diagnosis of lung cancer.     

Telomerase activity in benign and malignant cytologic fluids

BACKGROUND: Telomerase is a ribonucleoprotein that maintains telomeric base pair repeats at the ends of mammalian chromosomes during DNA replication. Telomerase is expressed in various human tumors, some normal tissues, and immortalized cell lines. The assay of telomerase activity has potential as an adjunct for cancer detection in cytologic fluids. METHODS: Twenty-four unfixed cytologic fluids, including 13 ascitic fluids, 7 pleural fluids, 3 pelvic washings, and 1 bronchial washing, were prepared for polymerase chain reaction (PCR)-based telomeric repeat amplification protocol (Oncor, Inc., Gaithersburg, MD). Telomerase activity was determined by PCR. The presence of a ladder of products with 6 base pair increments, separated by polyacrylamide gel electrophoresis and detected by phosphoimaging, was considered a positive result. Results were compared with cytologic evaluation of alcohol fixed, Papanicolaou stained smears. RESULTS: Of the 14 cytologically malignant specimens, 11 (79%) contained detectable telomerase activity. Two cytologically malignant samples could not be evaluated for telomerase activity due to the presence of inhibitory substances of PCR reaction. Of the 10 cytologically negative specimens, 1 (10%) was positive for telomerase activity; this specimen was from a patient with history of both endometrial and lung carcinomas. CONCLUSIONS: Telomerase activity can be detected in malignant cytologic specimens and thus has potential as a diagnostic adjunct in cytopathology.     

食管癌组织端粒酶活性的研究

目的：检测食管癌及周围非癌食管粘膜活性、了解食管癌发生可能的分子生物学基础。方法：应用ＴＲＡＰ法对３３例食管及１８例癌周正常食管粘膜进行端粒酶活性测定。结果：食管癌组织２８例（８４．８％）表达阳性,癌周组织２例（１１．１％）表达阳性。端粒酶活性表达与食管的性别,民族,肿瘤部位,分化程度,有无淋巴结转移等似乎无关。结论：端粒酶活化与食管癌发生有关,并可为食管肿瘤早期诊断与治疗提供可靠的指标与线索     

采用改进方法检测肺癌组织端粒酶活性

用改进的ＴＲＡＰ法检测肺癌的端粒酶活性,探讨端粒酶活性与肺癌的关系。方法：用培养瓶替代液氮瓶收集标本,采用ＴＲＡＰ改进方法,内含３６ｐｂ质控模板预防假阴性,增加强物长度避免二聚体形成,且使反应一步完成,并用银染法显示检测结果,对４５例病理确诊的肺癌手术标本及一株肺癌细胞株的端粒酶活性进行检测。     

膀胱癌组织中端粒酶活性的检测及其意义

目的 研究端粒酶与膀胱移行细胞癌（TCC）生物学行为的关系。方法 方法 采用端粒酶重复序列扩增法及酶联免疫吸附性检测58例TCC组织、10例癌旁膀胱粘膜及10例正常膀胱粘膜组织的端粒酶活性。结果 TCC组织端粒酶阳性率为86．2（50／58）,不同病理分级、临床分期之间无显著性差异（P＞0．05）;癌旁组织中只有1例端粒酶阳性;正常组织中端粒酶均为阴性。结论 TCC组织中端粒酶活性阳性率明显高于正常膀胱粘膜组织及癌旁膀胱粘膜组织。各病理分级和临床分期之间端粒酶活性虽然无显著性差异（P＞0．05）,但端粒酶活性的强弱分布不同。端粒酶活性可作为TCC早期诊断、鉴别诊断及预测复发的肿瘤标志物之一。     

胆汁脱落细胞端粒酶活性检测在胆道恶性肿瘤诊断中的价值

背景与目的：已有学者对尿液,胰液,肠腔冲洗液,胸水等脱落细胞端粒酶活性进行了检测,探索其对恶性肿瘤的诊断价值。而胆汁脱落细胞端粒酶活性的检测尚少见报道。本文的目的是探讨胆汁脱落细胞端粒酶活性检测及其对胆道恶性梗阻的诊断价值。方法：应用TRAP银染法检测胆汁脱落细胞的端粒酶活性。结果：44例恶性肿瘤胆道梗阻病人获得的胆汁中有33例检测到端粒酶活性阳性,阳性率75．0％,而良性梗阻组19例中只有1例检测到端粒酶弱阳性,阳性率5．3％。胆汁脱落细胞端粒酶活性阳性率与淋巴结转移,远处转移以及分化程度等肿瘤临床病理学特征无关,但胰腺癌和胆管癌病人胆汁中端粒酶活性检测阳性率高。端粒酶活性检测与脱落细胞学检查对比结果显示：44例胆道恶性梗阻的病人中31例进行了胆汁脱落细胞学检查,结果只有3例癌细胞阳性,阳性率9．7％,这3例脱落细胞学检查阳性的胆汁脱落细胞端粒酶活性检测均为阳性。结论：TRAP银染法可有效检测到胆汁脱落癌细胞的端粒酶活性。端粒酶可以作为诊断恶性胆道疾病的分子标志物。胆汁中脱落细胞端粒酶活性检测可作为细胞学检查的补充诊断手段。     

检测诱导痰中端粒酶活性对肺癌的诊断价值

目的:探讨检测诱导痰中端粒酶活性对肺癌的诊断价值。方法:肺癌患者48例,肺部良性疾病患者26例,两组患者均收集常规痰和诱导痰标本,采用TRAP-ELISA法测定端粒酶活性。结果:肺癌组患者常规痰液中端粒酶活性阳性检出率为47·9%,诱导痰中端粒酶活性阳性检出率为75·0%,两者差异有统计学意义,P=0·0181。肺癌组两种方法的标本端粒酶水平均高于对照组;而不同病理类型的肺癌端粒酶活性水平差异无统计学意义,P=0·6717。结论:诱导痰技术无创、简便,检测诱导痰中端粒酶活性对肺癌的临床诊断有一定价值。     

肺癌患者支气管镜活检标本端粒酶活性的测定

目的 验证肺癌中端粒酶活性的存在,探寻支气管镜活检标本的端粒酶活性测定在肺癌诊断中的价值。方法 以端粒重复扩增技术（ＴＲＡＰ）分析手术标本及支气管镜活检标本的细胞提取液中端粒酶的活性,ＴＲＡＰ分析的结果与病理检查结果进行比较,结果 １０例肺癌手术标本中均检测到端粒酶活性。１０例手术获得的正常肺组织中没有检测到端粒酶活性。１１例肺癌支气管镜活检标本中有８例检测垤端粒酶活性。３例肺良性病变支气管镜活检     

Real-time telomerase assay of less-invasively collected esophageal cell samples

Genomic and proteomic efforts have discovered a complex list of biomarkers that identify human disease, stratify risk of disease within populations, and monitor drug or therapy responses for treatment. Attention is needed to characterize these biomarkers and to develop high-throughput technologies to evaluate their accuracy and precision. Telomerase activity is correlated with tumor progression, indicating cells that express telomerase possess aggressive clinical behavior and that telomerase activity could be a clinically important cancer biomarker. Traditionally, the detection of cancer has involved invasive procedures to procure samples. There is a need for less invasive approaches suitable for population- and clinic-based assays for cancer early detection. Esophageal balloon cytology (EBC) is a low-invasive screening technique, which samples superficial epithelial cells from the esophagus. Since telomerase activity is absent in superficial cells of normal esophageal squamous epithelium but is often present in superficial cells from dysplastic lesions and ESCCs, measuring telomerase activity in EBC samples may be a good way to screen for these lesions. The development of rapid real-time telomerase activity assays raises the possibility of extending such screening to high-risk populations. In this study, we evaluate the feasibility of using rapid Real-Time Telomerase Repeat Amplification Protocol (RTTRAP) for the analysis of NIST telomerase candidate reference material and esophageal clinical samples. The telomerase activity of eight EBC samples was also measured by capillary electrophoresis of RTTRAP products, RApidTRAP, and hTERT mRNA RT-PCR assays. These findings demonstrate the feasibility of using the RTTRAP assay in EBC samples and suggest that individuals from high-risk populations can be screened for telomerase activity.     

Significance of telomerase activity in the diagnosis of small differentiated hepatocellular carcinoma

Precise diagnosis of well-differentiated hepatocellular carcinoma (HCC) is sometimes difficult to establish. Telomerase activity was examined by telomeric-repeat-amplification protocol (TRAP) in 37 HCC nodules smaller than 3 cm in diameter, including 24 fine-needle-aspiration biopsy specimens, 22 non-tumor chronic-liver-disease tissues (9 chronic hepatitis and 13 liver cirrhosis) and 3 normal liver tissues. Telomerase activity was assayed by serially diluted samples and quantitated by using an internal telomerase assay standard (ITAS). Telomerase activity was detected in all HCC and in 11 of 22 non-tumor chronic-liver-disease tissues. Normal liver samples had undetectable telomerase activity. Cut-off level of telomerase activity for its practical usage in HCC diagnosis was tentatively set for 0.6 μg liver protein/assay at 10-cell equivalent activity of a gastric-cancer cell line, MKN-1. This level was twice the highest activity in non-tumor chronic liver disease therefore, telomerase activity in all non-tumor liver samples was below this level. The telomerase-positive incidence exceeding this cut-off level was 73% (11/15) in well-differentiated HCC, 94% (16/17) in moderately differentiated HCC and 100% (5/5) in poorly differentiated HCC. Well-differentiated HCC showed low positivity by other diagnostic markers. 21% by AFP, 0% by PIVKA-II and 13% by angiography. The detection of telomerase activity may thus be a useful additional tool for precise and early diagnosis of small differentiated HCC, even when diagnosis is inconclusive by conventional techniques. Int. J. Cancer 74:141-147, 1997. © 1997 Wiley-Liss, Inc.