幽门螺杆菌感染与长乐门区胃癌发生的关系

目的：探讨胃粘膜幽门螺杆菌(Helicobacter pylori,Hp)感染对细胞凋亡及EGFR、VEGF表达的影响,从而初步探讨HP感染与长乐市高发区胃癌发生的关系。方法：采用碱性品红染色法和血Hp抗体检测法检测Hp感染状况;原位末端标记法（TUNEL法）检测胃粘膜细胞凋亡指数;免疫组化S－P法检测胃粘膜EGFR及VEGF表达情况。结果：长乐市胃癌温室伦系列Hp现症感染率（Hp 率）为74．4％,以慢性萎缩性胃炎伴或不伴肠化（CAG－IM）组最高,为92．5％;细胞凋亡指数在CAG－IM组最高,为0．357,而胃癌组最低,为0．179;EGFR表达阳性率为51．35％,其中CAG－IM组阳性率最高,达75．05,慢性浅表性胃炎（CSG）组最低,为25．05,胃癌组为46．7％;VEGF在不典型增生（AH）组最低,为35．0％,在胃癌组最高,为65．0％,（P＜0．05）。VEGFg怼umP《0、05）。Hp感染使胃粘膜凋亡指数增高,EGFR表达增加,VEGF表达下降（P＜0．05）。结论:Hp感染与长乐市高发区胃癌发生相关,Hp感染影响了胃粘膜细胞EGFR、VEGF的表达及胃粘膜细胞的增生和凋亡,从而可能影响了胃癌演化系列发生、发展的动态过程。     

肺癌纤维支气管镜刷检标本端粒酶活性检测

背景与目的:年来研究表明,端粒酶激活在肺瘤发生中起重要作用,但端粒酶活性水平检测用于肺癌小标本研究尚不多。本实验旨在探讨纤维支气管镜检查刷检标本端粒酶活性检测对肺癌诊断的价值。方法：应用TRAP－PCR－ELISA方法和TRAP－PCR－银染法检测66例纤维支 管镜刷检标本端粒酶活性,并分析其对诊断肺癌的价值。结果：56例肺癌纤支镜刷检标本端粒酶活性测定,阳性检出率为78．6％（44／56）,高于细胞学检查阳性率46．4％（26／56）（P＜0．005）,两者联合实验诊断肺癌敏感性93．7％。中央型肺癌纤支镜刷检标本端粒酶活性阳性率82．5％（33／40）,高于周围型68．8％（11／16）,但差异无显著性（P＞0．05）。定性法测定端粒酶活性阳性率与定量法相近,但发现定量法检测ΔA值较低的定性检测为弱阳性,条带较少,且条带不清晰。结论：端粒酶活性是肺癌诊断的分子生物学标志物之一。与细胞学检查相结合可提高肺癌的早期诊断率。     

不同病变胃粘膜端粒酶活性及其亚单位的检测

目的：探讨端粒酶及其亚单位（ＴＰ１、ｈＴＲ、ｈＴＲＴ）在胃粘膜癌变过程中的作用。方法：端粒酶的检测采用端粒末端重复序列扩增技术（ｔｅｌｏｍｅｒｉｃ　ｒｅｐｅａｔ　ａｍｐｌｉｆｉｃａｔｉｏｎ　ｐｒｏｔｏｃｏｌ,ＴＲＡＰ）,端粒酶亚单位的检测采用ＲＴ－ＰＣＲ法。结果：端粒酶阳性检出率在慢性萎缩性胃炎、肠上皮化生、异型增生及胃癌中分别为２４．６％（１４／５７）、３８．９％（７／１８）、３７．５％（３／８）及９２．３％（６０／６５）,正常胃粘膜未检测到端粒酶活性,与以上各病变组织有显著性差异（Ｐ＜０．０５～０．０１）,而慢性萎缩性胃炎、肠上皮化生和异型增生组织中端粒酶阳性率亦明显低于胃癌组织（Ｐ＜０．０１）;端粒酶的表达与临床病理指标无相关性;ＲＴ－ＰＣＲ定性检测发现端粒酶亚单位ｈＴＲ和ＴＰ１在大多数胃粘膜中都有表达,而ｈＴＲＴ主要在胃癌及部分癌前组织中表达,且在胃癌中ｈＴＲＴ的表达与端粒酶活性之间具有明显的相关性（Ｐ＜０．０１）。结论：端粒酶不仅在胃癌组织中高表达,在部分癌前组织中也有表达。提示端粒酶在胃癌的发生过程中可能具有重要作用;端粒酶亚单位ｈＴＲＴ不仅可能作为胃癌的诊断指标,而且还可能作为胃癌基因治疗的靶     

胃癌发生过程中端粒酶活性测定及临床意义

目的：通过检测胃癌及癌前病变组织中端粒酶活性，探讨其在胃癌发生发展中的作用及临床意义。方法：采用SP免疫组化法检测40例胃癌和18例癌前病变（其中慢性萎缩性胃炎伴肠化生10例，非典型增生8例）端粒酶活性。结果：胃癌及癌前病变中端粒酶阳性率分别为92．5％（37／40）和33．3％（6／18），两者比较差异有统计学意义，P〈0．05。端粒酶激活与胃癌的分化程度、病理分期和淋巴结转移无明显相关性，P〉0．05。结论：端粒酶活性表达与胃癌的发生密切相关，利用免疫组化检测端粒酶活性可作为胃癌早期诊断的参考指标，并应对阳性病例进行跟踪随访。     

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。     

喉癌及癌前病变组织端粒酶活性检测

目的 探讨喉鳞状细胞癌及喉癌前病变组织的端粒酶活性表达及其在喉癌发生中的作用。方法 采用多聚酶链反应－酶联免疫吸附法（PCR－ELISA）对47例喉鳞状细胞癌组织、22例喉癌前病变组织、20例喉炎性息肉组织的端粒酶活性进行定量检测。结果 47例喉鳞癌组织中39例端粒酶呈阳性（83％）,而20例喉炎性息肉组织端粒酶均为阴性,两组具有显著性差异（P＜0．001）。22例喉癌前病变组织中有15例端粒酶表达阳性（68．2％）。结论 端粒酶在喉癌组织中有较高的表达,可作为喉癌的肿瘤标志物。在喉癌前病变组织中也有低水平的端粒酶活性表达,喉癌前病变组织端粒酶活性检测将有助于喉癌的早期发现和早期治疗。     

原发性肺癌端粒酶活性检测及其临床应用价值

目的：研究原发性肺癌组织端粒酶活性检测的临床应用价值。探讨其作为肺癌诊断和预后评价指标的可行性。方法：采用端粒重复序列扩增－酶联免疫吸附法（TRAP－ELISA法），对32例原发性肺癌手术切除组织及相应的癌旁组织进行端粒酶活性分析，并以7例肺良性病变作为对照。结果：32例肺癌组织端粒酶水平明显高于癌旁组织（P＜0．05），及良性病变组织（P＜0．001），肺癌组织中端粒酶阳性率为84．4％，癌 旁组织为9．4％，在良性组织中未检出端粒酶阳性，端粒酶活性随临床分期有升高趋势，伴淋巴结转移标本组中其阳性率明显高于非淋巴结转移组（P＜0．05），但与肿瘤病理组织学类型，分化程度等未显示有统计学差异。结论：端粒酶激活与肺癌的发生，发展密切相关，端粒酶可以成为一种用于肺癌辅助诊断，预后判断的重要标志物并在肺癌的基因治疗中发挥作用。     

急性白血病端粒酶活性检测及其临床意义

目的：研究急性白血病发生发展不同阶段的端粒酶活性水平，探讨端粒酶活性检测对急性白血病的临床意义。方法：选取初治、治疗中和完全缓解期共33例成人急性白血病病例，以缺铁性贫血（IDA）和骨髓形态学正常的非血液病患者作对照，采用改良TRAP－银染法及亚利恩凝胶成像系统分析检测其骨髓标本内的端粒酶活性水平并随访。结果：缺铁性贫血患者端粒酶阴性或低水平表达；急性白血病患者端粒酶活性明显增高（P＜0．05），阳性率达70％，白血病完全缓解后端粒酶活性水平下降（P＜0．05），完全缓解期端粒酶活性仍高于正常者往往预后不良。结论：端粒酶与急性白血病的发生发展密切相关。端粒酶活性的检测可以作为白血病诊断及病情监测的一个生物学指标，有希望成为一种监测微小残留病的新的生物学标记。对白血病的抗端粒酶疗法有潜在的价值。     

子宫颈病变中端粒酶活性的表达

目的：探讨宫颈糜烂，宫颈上皮内瘤样变（CIN）及宫颈癌中端粒酶的表达及其意义，方法：采用TRAP法检测30例宫颈糜烂，16例CIN组织，20例宫颈癌及20例正常宫颈组织中端粒酶活性。结果：宫颈糜烂者端粒酶活性阳性率为3．0％（1／30），CIN端粒酶活性阳性率为56％（9／16），20例宫颈癌组织中端粒酶阳性表达率为805（16／20），正常宫颈组织中端粒酶表达阴性。恶性肿瘤与正常或良性病变组织中端粒酶活性差异有显著性（P＜0．05）。结论：端粒酶的活化在宫颈癌的发生过程中起重要作用，端粒酶活性检测可能成为宫颈癌及宫颈癌前病变早期诊断诊断和病情监测的指标。     

Telomerase activity: comparison between fine-needle aspiration and biopsy specimens for the detection of tumor cells.

BACKGROUND: The purpose of the current study was to determine telomerase activity as a sensitive biomarker for the detection of malignant cells in fine-needle aspiration (FNA) specimens. METHODS: FNA specimens with parallel samples of fresh tumor tissue were obtained from surgical specimens after surgical excision. Using a polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) assay, telomerase activity was determined systematically in FNA specimens (n = 21) and corresponding available tissue biopsy specimens (n = 16) containing malignant cells. In addition to a case of myelolipoma, normal counterparts for 3 of 16 cancer cases, including both biopsy and FNA specimens, also were available for the determination of telomerase activity. RESULTS: Telomerase activity was observed in 14 of 16 of the FNA specimens (88%) and 15 of 16 of the corresponding biopsy specimens (94%). Telomerase activity was detected in both the biopsy specimen and the corresponding FNA specimen, with one exception (a case of mucinous adenocarcinoma of the cecum). In contrast, specimens from three normal tissue biopsies and FNA specimens of normal tissue adjacent to the malignant lesions, as well as the myelolipoma, exhibited no telomerase activity. It is interesting to note that both tissue biopsy specimens and FNA specimens from a patient with high grade sarcoma were negative for telomerase activity. The examination of hematoxylin and eosin-stained adjacent tissue biopsy sections or FNA smears revealed similar low populations of lymphocytes, including those cases that were negative for telomerase activity. There was agreement in the detection of telomerase activity between tissue biopsies and their corresponding FNA specimens in 15 of the 16 patients, indicating a 94% concordance rate (95% confidence interval, 70%, 98%). CONCLUSIONS: The results of the current study clearly suggest that the telomerase activity in FNA specimens was comparable to that of their corresponding biopsy specimens, and that this activity was associated with the presence of malignant cells. The TRAP assay has potential for use in the detection of malignant cells in FNA specimens, particularly cases in which the cytology is not characteristically malignant and/or is present in insufficient numbers. Cancer (Cancer Cytopathol) Copyright 1999 American Cancer Society.     

Progressive increase in telomerase activity from benign melanocytic conditions to malignant melanoma

The expression of telomerase activity and the in situ localization of the human telomerase RNA component (hTR) in melanocytic skin lesions was evaluated in specimens from sixty-three patients. Specimens of melanocytic nevi, primary melanomas and subcutaneous metastases of melanoma were obtained from fifty-eight patients, whereas metastasized lymph nodes were obtained from five patients. Telomerase activity was determined in these specimens by using a Polymerase Chain Reaction-based assay (TRAP). High relative mean telomerase activity levels were detected in metastatic melanoma (subcutaneous metastases = 54.5, lymph node metastases = 56.5). Much lower levels were detected in primary melanomas, which increased with advancing levels of tumor cell penetration (Clark II = 0.02, Clark III = 1.1, and Clark IV = 1.9). Twenty-six formalin-fixed, paraffin-embedded melanocytic lesions were sectioned and analyzed for telomerase RNA with a radioactive in situ hybridization assay. In situ hybridization studies with a probe to the template RNA component of telomerase confirmed that expression was almost exclusively confined to tumor cells and not infiltrating lymphocytes. These results indicate that levels of telomerase activity and telomerase RNA in melanocytic lesions correlate well with clinical stage and could potentially assist in the diagnosis of borderline lesions.     

Enhanced expression of telomerase activity in thymoma and thymic carcinoma tissues: a clinicopathologic study

BACKGROUND: Telomerase is a nucleoprotein complex that caps the physical termini of all eukaryotic chromosomes. Because most malignant cells and reproductive cells have telomerase activity, which elongates telomeric DNA, telomerase may play important roles in unlimited cell division acquisition of the malignant phenotype. The current study examined the relation of telomerase activity in thymoma and thymic carcinoma with the clinicopathologic features of these lesions. METHODS: Tissue specimens were surgically resected from patients with thymoma and thymic carcinoma. Telomerase activity was evaluated according to a modified telomeric repeat amplification protocol assay. Paraffin sections of tumor were immunostained by MIC2 antibody, a marker of immature T cells. RESULTS: Telomerase activity was detected in all thymic epithelial tumors. The activity (mean +/- SD; unit per microg protein) in thymoma (n = 17) was significantly higher than that in thymic carcinoma (n = 7) (431.8 +/- 400.1 vs. 68.8 +/- 39.8; P < 0.01). Telomerase activities in thymoma and thymic carcinoma were significantly higher than that in primary lung adenocarcinoma (33.5 +/- 39.2, n = 47), studied as a control (P < 0.01). In patients with thymoma, telomerase activity did not correlate with tumor stage according to Masaoka classification (P = 0.776). In patients with thymic carcinoma, however, telomerase activity positively correlated with tumor stage (P = 0.02). In thymoma, telomerase activity positively correlated with the ratio of induced lymphocytes according to Rosai's classification (P = 0.045). MIC2-positive lymphocytes were identified in all cases of thymoma (n = 12). In contrast, lymphocytes infiltrating thymic carcinoma did not react with MIC2. CONCLUSIONS: In thymoma, telomerase activity reflects the presence of immature T-cell lymphocytes in tumor tissue rather than tumor stage or malignant phenotype. In thymic carcinoma, telomerase activity derived directly from cancer cells may relate to tumor stage.     

p27和ILK在胃癌及癌前病变中的表达及意义

目的：探讨p27和ILK在胃癌及癌前病变中的表达及其与胃癌病理参数之间的关系。方法：用免疫组化技术（SP法）对正常胃黏膜（normal gastric mucosa,NGM）、慢性浅表性胃炎（chronic superficial gastritis,CSG）、慢性萎缩性胃炎（chronic atrophia gastritis,CAG）伴肠上皮化生、慢性萎缩性胃炎伴非典型增生各20例和胃癌（gastric carcinoma,GC）60例标本进行免疫组织化学染色,分析p27和ILK的表达与胃癌临床和病理的关系。结果：p27和ILK表达阳性率分别为：NGM组100％和5％,CSG组85％和10％,CAG伴肠化组70％和20％,CAG伴非典型增生组45％和30％,胃癌组38．3％和40％。胃癌组和CAG伴非典型增生组p27阳性率显著低于其他组（P〈0．05）,ILK阳性率则显著高于其他组（P〈0．05）。p27和ILK在胃癌中的表达与肿瘤分化程度、浸润深度及肿瘤临床分期相关,p27蛋白的表达还与有无淋巴结转移相关。p27和ILK在胃癌中的表达呈显著负相关（r=-0．768,P〈0．05）。结论：检测胃癌组织中p27和ILK的表达有助于判断肿瘤的进展程度,两者联合检测有助于判断肿瘤预后。     

胃癌及癌前病变组织中TGFα和cyclin E的表达及两者关联性分析

背景与目的:研究发现,转化生长因子α(transforminggrowthfactoralpha,TGFα)或细胞周期素E(cyclinE)表达增高与肿瘤的发生、发展关系密切。但在胃癌前病变中的表达报道较少;两者表达的关联性分析未见报道。本研究旨在检测TGFα和cyclinE在慢性浅表性胃炎、胃癌前病变和胃癌组织中的表达情况,分析两者表达的关联性。方法:用免疫组织化学方法,检测TGFα和cyclinE在上述组织中的表达,分析两者表达在不同病理组织中的差异,以及两者表达的关联性。结果:在慢性浅表性胃炎、肠上皮化生、不典型增生及胃癌组织中,TGFα表达的阳性率分别为15.1%、53.6%、51.7%和61.7%,cyclinE分别为7.5%、28.6%、37.9%和42.6%,两者在肠上皮化生、不典型增生和胃癌的表达均高于在慢性浅表性胃炎的表达(均P<0.05)。在中-高分化腺癌和低分化腺癌,TGFα表达的阳性率分别为41.7%和81.0%,cyclinE分别为16.7%和57.1%,两者在低分化腺癌的表达均高于在中-高分化腺癌的表达(均P<0.05)。在胃慢性炎症组织(含癌前病变)以及在胃癌组织,TGFα和cyclinE的表达均存在显著的关联(分别为P<0.001和P=0.005)。结论:在慢性浅表性胃炎、癌前病变和胃癌组织中,TGFα和cyclinE的表达随病变严重程度以及胃癌恶性程度的增高而增高;且两者表达存在显著的关联     

细针穿刺肿瘤组织端粒酶活性检测在乳腺癌诊断中的意义

目的 :探讨细针穿刺乳腺肿瘤组织端粒酶活性检测在乳腺癌诊断中的意义。方法 :用PCR ELISA法检测79例术前乳腺肿瘤穿刺活检标本和大体标本的端粒酶活性并与病理诊断进行对照。结果 :乳腺癌 65例 ,穿刺组织端粒酶阳性 5 7例 ,阳性率为 87 7% ;大体组织端粒酶阳性 5 4例 ,阳性率83 1% ;淋巴结有转移者端粒酶活性高于无淋巴结转移者 ;乳腺良性疾病 14例 ,端粒酶阳性 2例 ,阳性率 14 3 %。结论 :术前乳腺肿瘤穿刺组织端粒酶活性检测有利于乳腺肿瘤的早期诊断及鉴别诊断 ,可以间接了解乳腺癌的进展程度     

脂溢性角化病皮损端粒酶活性的表达及其临床意义

目的 了解脂溢性角化病皮损中端粒酶活性的表达情况,探讨端粒酶与脂溢性角化病发病的关系。方法 采用端粒重复序列扩增文件——酶标法(TRAP-ELISA)检测30例脂溢性角化病和15例恶性黑素瘤以及15例正常皮肤组织中端粒酶活性的表达情况。结果 脂溢性角化病皮损端粒酶活性的表达与恶性黑素瘤皮损及正常皮损比较,其差异均具有统计学意义(P＜0.05)。结论 脂溢性角化病皮损组织中端粒酶的阳性表达,提示端粒酶可能参与了脂溢性角化病皮损的增殖过程。     

Detection of gastric cancer and premalignant lesions by novel marker glycoprotein 87 using monoclonal antibody Adnab-9.

Gastric adenocarcinoma (GC), the second most frequent cancer in the world, is highly prevalent in Asia. A screening test for early-stage GC would represent a major advance in the management of this disease. Associated conditions such as chronic atrophic gastritis (CAG) with intestinal metaplasia are manifested by specialized intestinalized tissue that often includes Paneth cells. Adenoma-associated antigen glycoprotein 87 (GP87), defined by the Adnab-9 monoclonal antibody and shed into the gut, is associated with epithelial cells, some of which resemble Paneth cells. We evaluated the diagnostic value of GP87 for cancer and gastric premalignant lesions. One hundred and seven sections of normal, benign, and premalignant gastric mucosa and 79 sections of pericancerous tissue were evaluated for expression of GP87 by immunohistochemistry. Eighty-two patients with GC, 34 patients with benign chronic disease, 35 patients with premalignant conditions, and 80 normal controls were evaluated by ELISA for GP87 in feces and gastric juice. Adnab-9 immunostaining for GP87 was significantly positive in CAG and intestinal metaplasia (>77.8%), compared with 0% in normal controls (P < 0.05). Fecal GP87 was positive in 79.3% of patients with gastric cancer, including early-stage lesions, and in 84% of patients with CAG versus 10% of controls (P < 0.05). Positive proportions for each pathological group in tissue correlated with that in feces (r = 0.99; P < 0.02). GP87 is differentially expressed in premalignant gastric lesions that appear to be the origin for fecal GP87, which may be useful for early detection of GC.