膀胱癌尿脱落细胞端粒酶活性检测及其临床意义

目的检测尿脱落细胞端粒酶活性并探讨其临床意义。方法应用改良的端粒重复序列扩增（ＴＲＡＰ）银染方法,分别对膀胱癌组织、正常膀胱组织,以及膀胱癌患者和非尿路上皮肿瘤患者的尿脱落细胞、膀胱冲洗液进行端粒酶活性检测。结果１２例正常膀胱组织均无端粒酶活性,４８例膀胱癌组织中４４例（９１．７％）端粒酶阳性。膀胱癌患者尿液及膀胱冲洗液中脱落细胞端粒酶阳性率分别为８３．３％（４０／４８）和８７．５％（４２／４８）。１２例分化良好（Ｇ１级）膀胱癌患者中,尿液和膀胱冲洗液中脱落细胞端粒酶阳性率分别为７５．０％（９／１２）和８３．３％（１０／１２）。结论尿脱落细胞端粒酶活性检测敏感性高,可用于膀胱癌的早期诊断和术后随访。     

尿脱落细胞荧光原位杂交技术诊断膀胱癌的价值

本文分析染色体异常与膀胱癌相关性，尿脱落细胞荧光原位杂交技术诊断膀胱癌的价值。     

膀胱癌尿脱落细胞CK20表达的临床意义

目的　探讨尿脱落细胞CK2 0表达在膀胱癌诊断中的意义。　方法　应用免疫组化技术检测 4 0例膀胱癌患者和 2 0例非肿瘤患者尿脱落细胞CK2 0的表达。　结果　非肿瘤组尿脱落细胞无CK2 0表达 ,肿瘤组尿脱落细胞CK2 0表达率为 5 3% ,CK2 0与HE染色结合检测 ,阳性率达90 %。复发组尿脱落细胞CK2 0表达率 80 % ,明显高于初发组的 4 3% (P <0 .0 5 )。CK2 0表达率与肿瘤分级、分期无相关性。　结论　尿脱落细胞CK2 0表达检测特异性高 ,可用于膀胱癌的早期诊断和术后随访     

膀胱癌患者尿脱落细胞存活素表达的临床意义

目的　探讨膀胱移行细胞癌 (TCCB)患者尿脱落细胞存活素 (survivin)表达的临床意义。　方法　采用巢式RT PCR方法检测 5 6例TCCB、2 8例对照组 (其中 8例膀胱外泌尿系肿瘤患者、10例非肿瘤泌尿系疾病患者及 10例健康者 )尿脱落细胞存活素的表达。　结果　 5 6例TCCB患者中尿脱落细胞存活素阳性表达 5 1例 (91.1% ) ,T1表达率 90 .5 % (19/ 2 1) ,T2～ 491.4 % (32 / 35 ) ;G1表达率90 .6 % (2 9/ 32 ) ,G2 90 .9% (10 / 11) ,G3 92 .3% (12 / 13) ,存活素表达与临床分期和病理分级不相关 (P >均 0 .0 5 )。对照组 2 8例中仅 1例血尿患者存活素阳性表达 (3.6 % )。TCCB组与对照组存活素阳性率比较差异有显著性意义 (P <0 .0 1)。尿脱落细胞存活素表达诊断TCCB的敏感性为 91.1% ,特异性为96 .4 %。　结论　尿脱落细胞存活素检测诊断TCCB敏感性、特异性高 ,无创 ,可作为诊断TCCB的敏感指标。     

尿脱落细胞荧光原位杂交技术诊断膀胱癌的价值

本文分析染色体异常与膀胱癌相关性,尿脱落细胞荧光原位杂交技术诊断膀胱癌的价值。     

膀胱癌尿脱落细胞染色体畸变的研究及临床意义

为明确荧光原位杂交在检测膀胱癌患者尿液，膀胱冲洗液中脱落细胞间期核染色体数目畸变的临床应用价值，收集２２例膀胱癌患者的尿液，２１例膀胱冲洗液，分别做尿细胞学、流式细胞术及荧光原位杂交的检测。同时收集５例正常人，５例前列腺增生患者尿液及２例膀胱冲洗液标本作为对照。结果显示：膀胱癌患者尿液组中尿细胞学、流式细胞术、荧光原位杂交分析阳性率分别为２７．３％（６／２２），４５．５％（１０／２２）和５０．０％（１１／２２），三者联合阳性率达６８．２％。膀胱冲洗液中阳性率则分别为２０．０％（４／２０），４０．０％（８／２０），７１．４％（１５／２１）和７６．２％（１６／２１）。认为荧光原位杂交技术检测膀胱癌患者尿液、冲洗液脱落细胞染色体畸变，可作为膀胱癌诊断的一项重要方法，并可能在术后监测复发以及预后判断中具有重要的临床意义。     

PCR-SSCP银染色法检测膀胱癌患者尿脱落细胞P53基因突变

采用ＰＣＲＳＳＣＰ银染色法检测５８例膀胱移行细胞癌患者尿脱落细胞Ｐ５３基因５８外显子突变。结果发现健康者和非膀胱癌患者的尿脱落细胞均无Ｐ５３基因突变，１９例膀胱癌患者尿脱落细胞存在Ｐ５３基因突变，其中Ⅰ、Ⅱ、Ⅲ级膀胱癌分别为０％（０／１６），２５％（７／２８）和８６％（１２／１４），Ｐ＜００１；表浅性（ＰＴｉｓ～Ｔ１）和浸润性膀胱癌（ＰＴ２Ｔ４）分别为１１％（４／３６）和６８％（１５／２２），Ｐ＜００１；初发和复发膀胱癌分别为９％（２／２２）和４７％（１７／３６），Ｐ＜００１；单发和多发膀胱癌分别为３１％（１２／３９）和３７％（７／１９），Ｐ＞００５。结论：ＰＣＲＳＳＣＰ银染色法是一种可靠、快速、实用的Ｐ５３基因突变检测方法。Ｐ５３基因突变主要发生在高级、期的膀胱癌，与膀胱癌的早期复发有密切关系，而与其多灶性无关。Ｐ５３基因突变的检测对预测膀胱癌的进展，复发的早期诊断具有重要的临床意义     

尿脱落细胞黏蛋白7 mRNA检测在膀胱癌诊断中的价值

目的 探讨尿脱落细胞黏蛋白7(Muc 7)mRNA检测在膀胱尿路上皮癌诊断中的价值.方法 采用RT-PCR方法检测52例膀胱癌患者和34例泌尿系非肿瘤患者尿脱落细胞中Muc7 mRNA表达情况,同时行尿细胞学检查,比较两种方法诊断膀胱癌的敏感性和特异性.膀胱癌患者52例,男30例,女22例.年龄41～87岁,平均65岁.TNM分期:Ta 1例、T1 22例、≥T2 29例.WHO分级:G1 18例、G2 14例、G3 20例.泌尿系非肿瘤患者34例,男23例,女11例.年龄30～75岁,平均58岁.结果 52例膀胱癌患者中Muc 7 mRNA检测阳性44例,敏感性为84.6%;34例非泌尿系肿瘤患者Muc 7 mRNA阴性29例、假阳性5例,特异性为85.2%,假阳性率为14.7%.膀胱癌患者尿脱落细胞检测阳性18例,敏感性为34.6%;非泌尿系肿瘤患者尿脱落细胞检测阴性31例、假阳性3例,特异性为91.2%,假阳性率为8.8%.尿Muc 7 mRNA检测诊断膀胱癌的敏感性优于尿脱落细胞学检查,差异有统计学意义(P<0.01);2种检测方法特异性及假阳性率比较,差异均无统计学意义(P>0.05).结论 尿Muc 7 mRNA检测诊断膀胱尿路上皮癌的敏感性优于尿脱落细胞学检查,可以作为膀胱癌的辅助检测指标.     

粘蛋白7在膀胱癌尿脱落细胞中的表达及意义

目的:探讨粘蛋白7(mucin7,MUC7)在膀胱移行细胞癌(TCC)诊断中的意义。方法:采用巢式逆转录聚合酶链反应(nested RT-PCR)方法检测MUC7mRNA在TCC患者尿脱落细胞中的表达。结果:68例TCC患者中有42例尿液MUC7表达阳性,敏感性为62%;MUC7表达与临床分期有关而与病理分级无关。对照组25例中有3例在尿液中MUC7为阳性表达,特异性为88%。结论:用RT-PCR的方法检测尿脱落细胞中的MUC7,具有较高的膀胱癌诊断价值。     

Immunostaining of cytokeratin 20 in cells from voided urine for detection of bladder cancer

PURPOSE: Cytokeratin 20 is a cytoskeletal protein expressed in colon and bladder cancer cells but only rarely in normal urothelium. Previous studies have shown that identification of RNA coded for cytokeratin 20 in urine samples using polymerase chain reaction is highly sensitive for detection of bladder tumors. We examined the efficacy of immunocytology on cells from voided urine samples using monoclonal antibodies against cytokeratin 20 for noninvasive detection of bladder tumors. MATERIALS AND METHODS: A total of 174 patients comprised the study, including 80 who were evaluated because of hematuria or irritative voiding symptoms and 94 who were examined during followup visits after resection of bladder tumors. Voided urine samples were obtained for immunocytology and cytopathology. Each patient underwent cystoscopy, and biopsies were done when a bladder tumor was detected or carcinoma in situ was suspected. Indirect immunoperoxidase staining was done on cytocentrifuge slides using a monoclonal antibody against cytokeratin 20. RESULTS: Bladder tumors were found in 87 patients. Immunocytology resulted in 81.6% sensitivity, 77% specificity and 80% accuracy. False-negative results occurred in 12 patients with superficial low grade tumors and in 4 with high grade invasive tumors. Cytopathology resulted in 51.7% sensitivity and 94.2% specificity. CONCLUSIONS: Immunocytology of the cytokeratin 20 antigen is significantly more sensitive than cytopathology for bladder tumor detection, especially low grade and low stage cancer. The results are not affected by inflammation or previous treatments with bacillus Calmette-Guerin. Immunocytology may miss high grade tumors and, therefore, cannot be used alone as a substitute for cystoscopy.     

从膀胱癌尿脱落细胞中提取RNA进行RT-PCR的研究

目的:建立从膀胱癌尿液脱落细胞中提取RNA进行RT-PCR研究的实验方法.方法:从20例膀胱癌患者尿液脱落细胞中提取RNA,采用RT-PCR技术扩增癌细胞中CD_(44)基因V_6～V_(11)外显子序列.结果:18例尿液标本均可被扩增出所设计的相应片段,并经Southern印迹杂交证实.结论:临床上采用此技术可从分子水平上对大批膀胱癌患者的癌基因表达产物进行无创伤性检测;为深入研究膀胱癌的发病机制、生物学行为和探讨简便、有效的无创伤性检测方法奠定了实验基础.     

荧光原位杂交技术对膀胱癌尿脱落细胞诊断价值的荟萃分析

目的通过荟萃分析比较荧光原位杂交技术（fluorescence in situ hybridization,FISH）和细胞学检测膀胱癌尿脱落细胞的灵敏度和特异度。方法从中国生物医学文献数据库、PubMed等专业数据库中收录的有关FISH、细胞学与膀胱癌尿脱落细胞检测相关分析文献中,纳入符合条件的文献,应用Revman5．0进行荟萃分析。结果HSH用于诊断膀胱癌尿脱落细胞的灵敏度为78．1％（0R值为4．98,95％CJ：3．63～6．83）,显著优于细胞学的46．3％（P〈0．00001）;HSH检查的特异度为93．9％（OR值为0．41,95％CI：0．193-0．930）,稍低于细胞学检查的96．3％（P=0．03）。结论荟萃分析肯定了FISH用于膀胱癌尿脱落细胞检测的价值,其灵敏度高,特异度接近细胞学检查。     

Flow cytometric study comparing paired bladder washing and voided urine for bladder cancer detection

Paired bladder washings and voided urines from bladder cancer patients were compared as sources of exfoliated cells for detection of bladder carcinoma by flow cytometry (FCM). Bladder specimens fixed in 25% ethanol within sixty minutes of collection were found to be superior to unfixed bladder specimens. The percentage of specimens with good DNA resolution was greater for bladder washings (67% unfixed, 90% fixed) than voided urines (41% unfixed, 66% fixed). There was no difference in DNA resolution between specimens that remained unfixed less than one day, one day, or two days suggesting that the cells undergo the majority of degradation within a critical period soon after collection. Once fixed, there was no difference in DNA resolution for up to nineteen days, which suggests the feasibility of specimen transport to central FCM laboratories. Eighteen percent of unfixed bladder washings and 33 percent of unfixed voided urine specimens contained an insufficient number of cells (less than 5,000) at the time of analysis compared with 6 percent bladder washings and 17 percent voided urines fixed in 25% ethanol. Flow cytometry and cytology results were concordant in 28/43 (65%) of fixed bladder washings and 9/13 (69%) of fixed voided urine. Voided urine was unreliable in providing consistent FCM data due to the high number of specimens with poor resolution or insufficient cells and is not recommended as a substitute for bladder washing when screening high-risk populations or monitoring patients with past history of bladder cancer.     

尿脱落细胞微卫星改变分析在膀胱肿瘤诊断中的应用

目的 选择6个微卫星位点，了解其在膀胱肿瘤诊断中的敏感性和特异性。方法 以外周血白细胞作为自身正常对照，检测31例膀胱肿瘤患肿瘤组织、尿脱落细胞以及10例正常人尿脱落细胞中的微卫星改变情况。结果 应用所选6个位点，诊断膀胱肿瘤具有90．3％的敏感性和100％的特异性，显高于尿细胞学检查的敏感性(10．3％)。结论 尿脱落细胞微卫星改变分析是一种无创、敏感、特异的诊断膀胱肿瘤的分子学方法。     

Microsatellite alterations in human bladder cancer: detection of tumor cells in urine sediment and tumor tissue

OBJECTIVES: Bladder cancer is the result of clonal expansion of cancer cells in which multiple genetic alterations have occurred. Loss of heterozygosity (LOH) studies have demonstrated that alterations in microsatellite regions are common in bladder cancer. This observation offers the possibility of early tumor detection by examining the DNA of urinary sediment.METHODS: We investigated alterations of 17 microsatellite loci in urinary bladder carcinomas of different stages and grades. Per locus, 19-30 specimens were evaluated. DNA was isolated from tumor specimens, urinary sediment and peripheral blood lymphocytes. DNA fragments of 17 microsatellite loci were amplified by PCR and analyzed for genomic alterations.RESULTS: Microsatellite alterations were detected in tumor tissue and urine sediment from 27 out of 31 patients (87%). Urine sediment analysis alone proved positive in 24 out of 31 patients (77%). The type of lesions most frequently detected was LOH (74% of all alterations), followed by length alteration (24%) and additional alleles (2%). On average, the alteration frequency was 22% per locus. The loci at chromosomes 9 and 18 proved most informative. No alterations were found in grade I tumors. The study revealed a correlation between microsatellite alterations and the respective grades and stages of the tumors. Average alteration frequencies per locus were: 27.4% in grade III versus 19.3% in grade II tumors, 26.5% in invasive versus 12.3% in superficial tumors.CONCLUSIONS: Our results demonstrate that microsatellite alterations are common in bladder cancer and that analysis of genomic instabilities in urine samples should be further evaluated as a method for bladder cancer screening in a high-risk group. Especially, when a set of microsatellites is used that shows a high probability of detecting alterations and allows easy handling, this could be an alternative or a completion to currently available methods.