联合检测尿液Survivin浓度及脱落细胞端粒酶活性在膀胱移行细胞癌中的诊断价值

目的：探讨尿液中Survivin浓度检测联合尿脱落细胞端粒酶活性检测在膀胱移行细胞癌诊断中的应用价值。方法：收集膀胱移行细胞癌患者尿液64例及非膀胱癌对照组患者尿液42例,应用ELISA方法检测各尿液标本中Survivin浓度及脱落细胞端粒酶的活性。结果：尿液中Survivin在膀胱癌诊断中的敏感度为82.8%,特异度为85.7%;尿脱落细胞端粒酶活性检测对膀胱癌诊断的敏感度为84.4%,特异度为69.0%;两种方法联合应用,两者均为阴性时诊断为阴性,否则为阳性,诊断膀胱癌的敏感度为95.3%,特异度为64.3%。结论：应用ELISA方法对尿液中Survivin浓度及脱落细胞端粒酶活性联合检测可显著提高膀胱移行细胞癌诊断的敏感度,而特异度变化不大,因此,两种指标联合检测较单一检测对膀胱移行细胞癌诊断更有价值。     

尿脱落细胞端粒酶活性检测在膀胱癌中的诊断价值

目的：探讨通过检测尿脱落细胞端粒酶活性早期诊断膀胱癌。方法：采用改进的银染－TRAP（端粒重复序列扩增）法,分别对45例膀胱移行细胞癌患者,10例膀胱良性病变患者及8例正常人尿脱落细胞进行了端粒酶活性检测。结果：24例G1级（分化良好）的膀胱癌患者,端粒酶活性强阳性12例,弱阳性6例（阳性率75．0％）,21例G2,G3级膀胱癌患者,强阳性16例,弱阳性3例（阳性率94．5％）,膀胱良性病变患者及正常人尿脱落细胞只有1例端粒酶活性呈弱阳性,其余全部阴性。结论：尿脱落细胞端粒酶活性检测特异性强,敏感性高,与病理分类,组织分级等无明显关系,提示有可能成为膀胱癌临床早期诊断指标之一。     

尿透明质酸含量诊断膀胱癌的临床研究

目的　探测尿液透明质酸 (HA)含量检测在膀胱移行细胞癌诊断中的意义并研究放免方法代替ELISA方法检测尿液中HA含量的可能性。方法　同时使用ELISA和放免两种方法对 49例膀胱移行细胞癌病人、12例良性膀胱肿瘤病人、30例BPH和泌尿系结石病人和 2 0例正常人尿液中HA含量进行检测。结果　ELISA方法和放免方法测得各组尿液HA值结果相近 (P >0 .0 5 )。以放免方法所得结果进行分析 ,膀胱移行细胞癌病人组尿液HA含量是良性膀胱肿瘤病人组、泌尿系非肿瘤性疾病病人组和正常成人组的 2 -4倍 ,差异性极显著 (P <0 .0 1)。以对照组尿液HA水平上限137.5ng/mg( 113 .6 + 2 3.9=137.5ng/mg )作为阳性界值时 ,尿液HA含量检测诊断膀胱移行细胞癌的敏感度为 91.8% ,特异度为 91.9% ,与尿脱落细胞学相比 (敏感度为 48.9%、特异度为 95 .2 % ) ,敏感度差异有极显著意义 (P <0 .0 1) ,特异度差异无统计学意义 (P >0 .0 5 )。结论　尿液HA含量检测是一种简单、无创、价廉、结果客观、敏感度和特异度均较高的诊断膀胱移行细胞癌的方法 ;价格更为低廉的放免法可代替ELISA方法进行尿液HA含量的检测。     

膀胱移行上皮癌尿脱落细胞端粒酶活性检测

[目的]检测尿脱落细胞端粒酶活性表达对膀胱移行上皮癌的诊断价值.[方法]应用PCR为基础的端粒重复片段扩增方法(TRAP-Hyb Kit)分别检测52例膀胱移行上皮癌患者及10例泌尿生殖系良性疾病患者尿脱落细胞端粒酶活性表达情况,并与尿脱落细胞学检查结果进行比较.[结果]52例膀胱移行上皮癌患者中有34例的自排尿标本中可检测出端粒酶活性表达(65.4%);尿脱落细胞学检查阳性例数为11例(11/30,36.7%);两者比较差异有显著性(P＜0.05).泌尿生殖系良性疾病的尿脱落细胞端粒酶的阳性率为10.0%.但端粒酶活性与肿瘤的分级无相关性.[结论]尿脱落细胞端粒酶活性检测作为无创性的检测方法,可作为膀胱移行上皮癌早期诊断、术后监测的一种有效手段.     

端粒酶活性检测在膀胱肿瘤早期诊断中的应用

目的：探讨检测端粒酶活性在膀胱癌早期诊断中的临床意义。方法：采用TRAP-PCR-ELISA法检测32例膀胱癌患者尿液和膀胱冲洗液中脱落细胞、膀胱癌组织、20例正常膀胱组织及14例非膀胱肿瘤患者尿液脱落细胞中端粒酶活性，并行尿脱落细胞学检查。结果：32例膀胱癌患者尿液脱落细胞、膀胱冲洗液脱落细胞、膀胱癌组织中端粒酶活性阳性率分别为65．6％（21／32）、71．9％（23／32）和84．0％（27／32），20例正常膀胱组织端粒酶活性均为阴性，14例非膀胱肿瘤患者尿液中1例端粒酶活性阳性。端粒酶活性阳性表达与肿瘤的分级、分期之间差异无明显相关性（P〉0．05），敏感性明显高于脱落细胞病理学检查。结论：尿液、膀胱冲洗液脱落细胞端粒酶活性测定敏感性较高．可用于膀胱癌的早期诊断。     

尿液中核基质蛋白22联合端粒酶活性检测诊断膀胱癌的实验研究

目的:探讨尿液中核基质蛋白22(NMP22)联合端粒酶活性检测在膀胱癌诊断中的价值,寻找一种敏感特异的无创性方法诊断膀胱癌.方法:收集60例膀胱癌和40例非膀胱癌患者新鲜尿液,用化学发光分析法检测尿液中NMP22水平,TRAP-PCR-ELISA法检测尿脱落细胞端粒酶活性,并同时行尿脱落细胞学检查,对比三者对膀胱癌诊断的敏感性和特异性.结果:尿液中NMP22对膀胱癌诊断的敏感性83.3％,特异性43.3％;端粒酶敏感性82％,特异性90％;细胞学检查敏感性30％,特异性100％.NMP22联合端粒酶活性检测,敏感性96％,特异性78％.结论:联合检测NMP22、端粒酶活性有较高的敏感性和特异性,具有一定临床诊断、随访价值.     

尿细胞角蛋白检测与尿脱落细胞学检查在膀胱癌诊断中的价值

目的:探讨尿细胞角蛋白检测与尿脱落细胞学检查在膀胱移行细胞癌诊断中的价值。方法:136例怀疑膀胱癌者,进行尿细胞角蛋白8和18的含量(UBC值)。检测与尿细胞学检查,其中87例经组织学证实为膀胱移行细胞癌。比较两者诊断膀胱癌的敏感性和特异性。结果:尿细胞角蛋白的敏感性为70.1%,特异性为73.3%;尿细胞学的敏感性为42.5%,特异性为83.7%。尿细胞角蛋白在膀胱癌不同分级和分期中的敏感性优于尿细胞学(P<0.05)。结论:尿细胞角蛋白的检测在早期诊断膀胱癌方面优于尿细胞学检查,可作为膀胱癌的早期检测指标。     

癌性体液细胞中端粒酶活性的研究及意义

 目的　研究癌性体液脱落细胞的端粒酶活性及在良恶性体液的鉴别诊断中的应用价值。方法　应用 PCR- ELISA方法检测体液脱落细胞中端粒酶活性 ,并将检测结果与细胞学检查进行比较。结果　癌性胸、腹水端粒酶阳性率为 61 .1 1 % (33/54) ;其中 ,肺癌胸水脱落细胞端粒酶阳性率为 70 % (2 1 /30 ) ,而形态学阳性率为 53.33% (1 6/30 )。膀胱癌尿液端粒酶阳性率为 83.64%(46/55) ,而形态学阳性率为 52 .72 % (2 9/55)。结论　端粒酶活性检测在癌性体液的诊断和鉴别诊断方面有重要的应用价值。     

BTA检测在膀胱癌诊断中的价值

目的BTA检测与尿脱落细胞学检查结果比较,以明确在膀胶癌诊断中的应用价值.方法 收集1996年12月～1997年1月经膀胱镜及病理学检查确诊为膀胱乳头状移行细胞癌的病人共47例.每一例于膀胱镜检查前连续留取三次晨尿,行尿脱落细胞学检查.最后一次标本同时行BTA检测及尿液常规化验.结果BTA检测膀胱癌的敏感度为70.2%(33/47),尿脱落细胞学检查敏感度为25.5%(12/47),两者有非常显著性差异(P<0.001).共有8例(17.0%)患者两种检查结果均为阳性,10例(21.3%)患者两种检测结果均为阴性.另外,BTA检测对T1期膀胱癌患者的敏感度明显高于尿脱落细胞学检查,其结果分别为76.0%(19/25)和12.0%(3/25),统计学分析示有非常显著性差异(P<0.01).结论 BTA检测是一种有价值的膀胱癌诊断辅助措施,且使用方便,检测迅速,无创伤性,便于临床开展.     

膀胱移行细胞癌尿液脱落细胞中Livinα的表达及意义

目的:研究膀胱移行细胞癌(BTCC)尿液脱落细胞中Livinα的表达及意义.方法:留取49例BTCC患者、21例其他系统疾病患者和15例正常健康成人的新鲜尿液,离心收集脱落细胞,以逆转录多聚酶链反应(RT-PCR)检测尿液脱落细胞中Livinα的表达,并行尿脱落细胞学检查.结果:49例BTCC患者尿脱落细胞中有40例检测出Livinα表达,而21例其他系统疾病患者和15例正常健康成人的尿脱落细胞中均未检测出Livinα的表达.以RT-PCR 方法检测膀胱癌患者尿液脱落细胞中Livinα的敏感性为81.6%,特异性为100%.尿脱落细胞学检查的敏感性和特异性为16.3%和100%,两种方法之间存在显著性(P<0.05).结论:初步的试验结果显示,RT-PCR法检测膀胱癌患者尿液脱落细胞中Livinα的方法比细胞学检查灵敏度高,可能成为诊断膀胱癌的无创性方法.     

Diagnostic Role of Survivin in Urinary Bladder Cancer

Background: Early diagnosis of carcinoma of bladder remains a challenge. Survivin, a member of theinhibitor of apoptosis (IAP) protein family, is frequently activated in bladder carcinoma. The objective of thisstudy was to investigate urinary survivin as a marker for diagnosis of urinary bladder. Materials and Methods:We examined urinary survivin concentration in 28 healthy individuals, 46 positive controls and 117 cases ofhistologically proven TCC prior to transurethral resection, using ELISA, and compared values with findings forurinary cytology. Results: Survivin was found to be significantly higher in the cancer group (P<0.05). A cut offvalue of 17.7 pg/ml was proposed, with an approximate sensitivity of 82.9% and specificity of 81.1% (P<0.0001),whereas urine cytology had a sensitivity of 66.7% and a specificity of 96.0%. Conclusions: Urinary survivin canbe used as a non-invasive diagnostic biomarker for TCC bladder, both for primary and recurrent disease.     

尿液核基质蛋白22在膀胱移行上皮癌筛检中的价值

目的：评价尿核基质蛋白22(NMP22)在膀胱移行上皮癌筛检中的意义。方法：采用酶联免疫法检测28例膀胱移行上皮癌、25例泌尿系良性疾病和10例泌尿系非移行上皮癌患者尿NMP22水平，并与尿脱落细胞学检查进行比较。结果：膀胱移行上皮癌患者尿NMP,,中位值为66．5u／ml，明显高于泌尿系良性疾病和非移行上皮癌，与肿瘤分期、分级、数目无关，以10u／ml为临界，诊断膀胱癌敏感性为85．7％，特异性为60％，尿脱落细胞学检查敏感性为32．1％，特异性为100％。结论：尿NMP22检测膀胱移行上皮癌敏感性、特异性均较高，可用于膀胱移行上皮癌的筛选和术后随访。     

Telomerase activity in bladder cancer, bladder washings and in urine

With only a few exceptions, the ribonucleoprotein telomerase has been found in malignant, but not in benign tissues. Telomerase is thus a potentially new diagnostic marker. Carcinoma of the urinary bladder is the most frequent malignant tumor of the urinary tract and, after prostatic carcinoma, the second most common malignancy of the genitourinary system. In order to evaluate the diagnostic capabilities of telomerase in bladder carcinomas, four cell lines derived from human urothelial carcinomas of the bladder, 75 tissue samples from bladder carcinomas, eight tissue samples of normal bladder urothelium, 40 bladder washings and 30 urine samples were examined for telomerase activity. The four cell lines derived from urothelial carcinoma of the bladder (F975, 582, SCaBER, UM-UC-3) all exhibited high telomerase activity and were thus used as positive controls. Telomerase activity was found in nearly all (96%) tissue samples obtained from histologically confirmed urothelial carcinoma of the bladder. None of the normal tissue samples examined showed telomerase activity. Telomerase activity was similarly found in 73% of bladder washings in patients with histologically confirmed bladder carcinoma. There were no false positive results. The determination of telomerase activity in bladder washing samples thus represents a new diagnostic method for detection of tumor cells in rinsing media. Because of the early inactivation or degradation of telomerase there was no detection of the enzyme in native urine in the present study.     

The clinical relevance of urine-based markers for diagnosis of bladder cancer

The aim of the present study was to evaluate the diagnostic relevance of urinary fibronectin (FN), telomerase (RTA), and cytokeratin 20 (CK20) mRNA in comparison with voided urine cytology (VUC). The study included 132 patients with bladder cancer, 60 patients with benign bladder lesions, and 48 healthy individuals. All were subjected to urine cytology, estimation of fibronectin by ELISA, RTA by TRAP, and CK20 mRNA by conventional RT–PCR in urothelial cells from voided urine. The best cutoff point for FN was determined by receiver operating characteristic curve (41.7 ng/mg protein) revealed the highest sensitivity for malignant (80%) followed by the benign (70%) than the healthy individuals (4.1%) at P < 0.001. Also, RTA and VUC showed significant difference among the three investigated groups (P < 0.001). The overall sensitivity (89.3%) and specificity (98.4%) were the highest for CK20 mRNA. Combined sensitivity of VUC with FN, RTA, and CK20 mRNA together (98.4%) was higher than either the combined sensitivity of VUC with any of them or than that of the biomarker alone. Accordingly, when the diagnostic efficacy was considered, CK20 mRNA had the highest sensitivity and specificity compared to all investigated markers.     

Sandwich ELISA for detecting urinary Survivin in bladder cancer

Objective

Survivin as a tumor marker in the diagnosis of bladder cancer has not been completely confirmed yet and there are few reports about using Survivin enzyme-linked immunosorbent assay (ELISA) kit to detect the urine of bladder cancer patients. This study aimed to develop a Survivin ELISA and validate its value in the detection of bladder cancer.

Methods

Through square matrix titration, different combinations of coating antibody and detecting antibody, a Survivin ELISA was constructed. This assay was evaluated according to intra-assay precision, inter-assay precision and minimum detectable dose (MDD). Survivin levels were detected and analyzed in 102 bladder cancer patients and 102 healthy people by established ELISA. Then cutoff value was defined according to the analysis of receiver operating characteristic (ROC) curve. The sensitivity and specificity of detection were calculated on the basis of cutoff value to diagnose bladder cancer patients. Furthermore, the value of Survivin expression detected by ELISA among different clinicopathological characteristics of patients was also compared.

Results

Through optimization of different conditions, intra-assay precision was 8.39%, inter-assay precision 8.57% and MDD 0.0625 ng/mL in this assay. When the optical density at 450 nm (OD₄₅₀) was 0.09, it could get the optimized diagnostic cutoff value. According to this value, the sensitivity and specificity of diagnosis in bladder cancer patients were 70.6% and 89.2%, respectively. The associations between patients’ clinical variables and OD₄₅₀ were not significant except tumor numbers in patients.

Conclusions

This experiment has preliminarily developed a Survivin ELISA and confirmed Survivin as a biomarker which owned a practical and significant value in the diagnosis of bladder cancer.Key Words: Survivin, bladder cancer, enzyme-linked immunosorbent assay (ELISA), tumor marker, diagnosis     

Urinary cytology and bladder cancer. The cellular features of transitional cell neoplasms

Urinary cytology is increasingly accepted as a diagnostic tool in the detection and follow-up of patients with bladder cancer. Its potential value has been reduced, however, by the relative inexperience of most pathologists in the examination of urinary specimens, and by the lack of cellular criteria specifically reflecting the morphology of low-grade papillary and flat lesions of bladder epithelium. The cytologic features of urothelial lesions, including papillary transitional cell carcinomas and flat urothelial dysplasias have been studied in both experimental systems and clinical situations and their application to a selected patient population is presented. Overall, the cytohistologic correlation for patients with bladder cancer was 92%. Positive cells reflecting the morphology of the tumor occurred in 62% of patients with grade I transitional cell carcinomas, and cells suspicious for malignancy were identified in an additional 14% of these individuals. Using the criteria presented, a positive cytology can correlate with a papillary grade I bladder tumor, and should not necessarily indicate the presence of another neoplasm. Dysplastic cells in cytologic specimens are often identified in patients having urothelial dysplasia as the most serious bladder lesion, but the cytologic diagnosis of dysplasia may represent an under-interpretation of a low-grade papillary bladder tumor. The cells of urothelial neoplasms, including low-grade transitional cell carcinomas and dysplasias, differ morphologically both from normal and reactive/reparative elements, and can be detected in cytologic samples. The changes are often subtle and require experience and a cautious approach for accurate interpretation.     

尿CYFRA21-1、端粒酶、VEGF联合检测膀胱移行细胞癌的价值

 目的　探讨反映肿瘤发生发展不同阶段的尿瘤标联合 ,检测膀胱移行细胞癌 (BTCC)的价值。方法　 4 5例BTCC患者 ,同时分别以抗体夹心ELISA方法检测尿CYFRA2 1 1、VEGF、TRAP方法检测尿端粒酶。联合检测的灵敏度与单一瘤标及尿细胞学检测结果比较 ,χ2 检验。结果　尿CYFRA2 1 1、端粒酶、VEGF诊断BTCC的灵敏度分别为 82 .2 %、77.8%、84 .4 % ,与尿细胞学 (44 .4 % )比较 ,均有显著性差异 (P <0 .0 0 5 ) ;联合瘤标诊断BTCC的灵敏度为 95 .6 % ,与尿细胞学 (P <0 .0 0 5 )