膀胱癌组织中血管内皮生长因子的表达及与血管生成的关系

目的 探讨血管内皮生长因子（ＶＥＧＦ）在原发性膀胱移行细胞癌中的表达及其与膀胱癌血管生成之间的关系。方法 免疫组织化学技术检测６８例原发性膀胱移行细胞癌和７例正常膀胱组织中血管内皮生长因子的表达,测定４０例浸润性膀胱癌的微血管密度,并分析ＶＥＧＦ和ＭＶＤ间以及它们与膀胱癌病理分级和临床分期间的关系。结果 ＶＥＧＦ在正常膀胱中不表达或低表达,而在膀胱表达较强;ＶＥＧＦ表达及ＭＶＤ值均与肿瘤的病理分级     

Caveolin-1在肝细胞癌表达及与肿瘤血管生成的关系

目的：探讨caveolin-1与肝细胞癌（HCC）侵袭转移的关系及其可能的机制。方法：应用实时PCR方法检测伴肝内转移的 HCC组织、癌旁组织、肝硬化组织、正常肝组织中 caveolin-1 mRNA的表达;用Western blot方法检测伴肝内转移的 HCC组织、癌旁组织中caveolin-1蛋白的表达;用免疫组化方法检测75例HCC组织中caveolin-1,血管内皮生长因子（VEGF）,CD34,α-平滑肌肌动蛋白（α-SMA）的表达,分析caveolin-1,VEGF的表达以及肿瘤微血管密度（MVD）,非成对动脉（UA）计数与临床病理因素的关系。结果：在伴肝内转移的HCC组织中,caveolin-1 mRNA表达明显高于癌旁组织、肝硬化组织、正常肝组织（均P<0.05）,其蛋白表达明显高于癌旁组织;caveolin-1表达水平的升高与肿瘤转移有关（P<0.05）,且caveolin-1的表达与VEGF表达及MVD和UA计数呈正相关（r=0.293,P=0.011;r=0.361,P=0.001;r=0.388,P=0.001）。结论：caveolin-1表达升高促进HCC的侵袭转移,其机制可能是通过诱导HCC表达VEGF,促进肿瘤新生血管生成而实现的。     

溶酶体相关基因在骨肉瘤中的表达和功能

目的 寻找可能的骨肉瘤血清蛋白标志物,探讨溶酶体相关基因在骨肉瘤中的表达和功能.方法 分别利用基因芯片和激光解析离子化飞行时间质谱(SELDI-TOF-MS)技术筛选3个骨肉瘤细胞株和27例骨肉瘤患者血清中差异表达的基因和血清蛋白质峰.采用Link-test统计学方法 筛选得到骨肉瘤血清标志蛋白,采用MATLAB软件对653个差异表达的标志蛋白进行功能聚类分析,并对溶酶体相关基因进行KEGG分析.结果 本实验在骨肉瘤细胞株中共筛选出653个候选骨肉瘤标志基因,在骨肉瘤患者血清中共筛选到9个差异表达的蛋白质谱峰.通过Link-test统计分析,共筛选到15个差异表达的骨肉瘤血清标志蛋白.骨肉瘤相关基因SCARB2、GNS和DNASE2等在溶酶体信号通路中显著富集.结论 溶酶体相关基因表达失调和溶酶体功能紊乱是骨肉瘤形成的原因之一,可以作为骨肉瘤的诊断标志物.溶酶体可以作为骨肉瘤治疗的靶细胞器.     

Expression of Maspin in non-muscle invasive bladder carcinoma: correlation with tumor angiogenesis and prognosis

OBJECTIVES: Maspin is a member of the serpin (serine protease inhibitor) family and has been shown to be a suppressor of tumor growth and an inhibitor of angiogenesis as well as metastasis in several types of tumors. We studied expression patterns of Maspin in pTa/pT1 urothelial carcinoma of the bladder and compared them with microvessel density (MVD) for two vascular markers (CD34 and CD105) and correlated the findings with clinical outcome. MATERIAL AND METHODS: We investigated tumor samples of 110 patients undergoing transurethral resection for pTa/pT1 bladder carcinoma (pTa, n=84; pT1, n=26; grade 1, n= 22; grade 2, n= 81; grade 3, n=7). Immunohistochemical studies were performed using the monoclonal antibodies, anti-human Maspin (NCL Maspin), anti-CD34 Class II and anti-CD105. Maspin expression level was classified according to the staining intensity (- to +++). The blood vessels (CD34) and specifically proliferating blood vessels (CD105) were counted as vessels per field (microvessel density, MVD). RESULTS: Of the 110 tumors, 27 showed a negative immunostaining for Maspin, 46 tumors stained +, 29 stained ++, and 8 stained +++. Maspin expression correlated inversely with CD34 reactivity. In tumors with loss of or only weak Maspin expression, the MVD for CD34 was 21.7 vessels per field, and 4.2 vessels per field for proliferating vessels (CD105), whereas Maspin-positive tumors had an MVD of 17.7 vessels per field (CD34), and of 6.0 vessels per field (CD105). Complete follow-up data are available in 92 patients. After a median follow-up of 25 months, 18 of the 92 patients (19.6%) had tumor recurrences. Tumors with decreased Maspin expression (-/+) had a shorter disease-free interval (23 months) than patients with stronger Maspin (++/+++) expression (29 months), whereas a Kaplan-Meier analysis and the log-rank test showed no significant difference in disease-free survival between the patients. CONCLUSION: The clinical importance of Maspin has been mainly investigated regarding tumor progression or metastasis. We found a decreased Maspin expression in a large portion of pTa/pT1 bladder tumors. Even if patients with decreased Maspin expression have a slightly shorter disease-free survival Maspin does not appear to be a promising prognostic marker.     

Ⅱ型环氧合酶在膀胱移行细胞癌中的表达及与肿瘤血管发生的关系

目的:研究Ⅱ型环氧合酶(Cox -2)在膀胱移行细胞癌中的表达,探讨它与膀胱癌病理分级及临床分期的关系及其临床意义,并研究它与血管内皮细胞生长因子(VEGF)及微血管密度(MVD)表达的关系,从而探讨Cox- 2在膀胱肿瘤血管发生中所起的作用。方法:应用免疫组化方法检测Cox- 2、VEGF及MVD在94 例膀胱移行细胞癌、12例膀胱良性病变和5例膀胱癌旁正常组织中的表达。结果:Cox- 2的表达随着膀胱癌分级分期的上升而呈上升趋势(P<0.05);在膀胱癌中,Cox 2 表达与VEGF表达( r=0.716, P=0.000)关系十分密切,与MVD表达存在明显相关性( r=0.458,P=0.000)。结论:Cox- 2在膀胱癌中均为高表达,它的表达参与了肿瘤的发生及恶性进展,并且与膀胱肿瘤新生血管发生有着密切关系。     

Plexin A1在胃癌中的表达及其与肿瘤细胞增殖和血管生成的关系

目的探讨PlexinA1在胃癌组织中的表达及其与肿瘤细胞增殖和血管生成的关系。方法应用RT-PCR检测20例胃癌患者的癌组织及胃切缘正常胃黏膜组织Plexin A1的mRNA表达;免疫组织化学方法检测50例胃癌组织和20例胃正常黏膜中PlexinA1、肿瘤细胞增殖指数Ki-67、血管内皮生长因子(VEGF)和第Ⅷ因子微血管密度(MVD)的表达。结果RT-PCR示胃癌组织中的PlexinA1mRNA的表达明显高于胃正常黏膜(0.71±0.37vs0.60±0.25,P<0.05)。胃癌组织中MVD随着PlexinA1的mRNA表达的增强(r=0.8736,P<0.01)和蛋白表达的增高而增高(P<0.01);Ki-67的表达也与PlexinA1存在明显的一致性(r=0.4851,P<0.01);而VEGF的表达与PlexinA1的表达无关。结论PlexinA1在胃癌发生发展中发挥重要作用,与调节血管生成和促进肿瘤细胞增殖有关。     

Tie2受体在人肝癌的表达及其与血管生成和肿瘤发展的关系

目的　探讨Tie2受体在肝癌血管生成中的作用。方法　检测CD34、Tie2受体在 2 2例人肝细胞性肝癌、8例门脉性肝硬化和 5例正常肝脏的表达 ,并对Tie2表达水平与肿瘤生物学行为的关系进行分析。结果　 (1 )CD34在正常肝脏无表达 ,在肝硬化肝脏偶见 ,在肝癌表达明显增加 (P <0 0 1 ) ;(2 )Tie2受体在正常肝脏无表达 ,在肝硬化肝脏偶见 (1 1 3± 8 7/HP) ,在肝癌微血管内皮大量表达 (52 4± 1 6 7/HP ,P <0 0 1 )。 (3)Tie2表达水平和肝癌直径、血管生成、门静脉侵犯相关 ,而与肿瘤包膜浸润、组织学分级、复发无关。结论　人肝细胞肝癌血管生成增加 ,Tie2受体和肝癌血管生成、肝癌大小、血管侵犯等相关 ,Angiopoietin /Tie 2可能参与肝癌血管生成的调节     

胃癌组织中Cox-2的表达与肿瘤血管生成及周围静脉血微转移的关系

目的 研究胃癌组织中Cox-2的表达及其与肿瘤血管生成和周围静脉血微转移的关系.方法 用免疫组织化学法检测42例胃癌组织中Cox-2的表达和微血管密度(MVD),用巢式RT-PCR法检测胃癌病人外周血癌细胞微转移.结果 本组有24例胃癌组织中Cox-2表达阳性.Cox-2阳性肿瘤的MVD计数显著大于Cox-2阴性者(P＜0.01).有13例周围静脉血中CK19mRNA表达阳性.Cox-2阳性胃癌病人外周血微转移阳性率显著高于Cox-2阴性者(P＜0.05).结论 胃癌组织中Cox-2表达的增加与胃癌组织血管化程度及周围静脉血微转移密切相关.     

VEGF在肾母细胞瘤中的表达及其与肿瘤血管形成和转移关系的研究

目的研究VEGF在肾母细胞瘤的表达及其与肿瘤血管形成和转移(包括淋巴结或/并肺转移等,下同)的关系。方法用免疫组化S-P法对25例肾母细胞瘤组织中血管内皮生长因子(VEGF)和肿瘤内微血管密度(MVD)进行检测。结果VEGF在肾母细胞瘤中阳性表达13例,阳性表达与MVD、肿瘤转移、临床分期均有显著性差异(P<0.05),而与病理类型无统计学意义(P>0.05)。结论VEGF的表达与肿瘤的血管形成和转移存在密切关系,其检测可作为判断肾母细胞瘤的转移和预后的指标。     

胃癌组织中 EphB4受体及其配体 EphrinB2的表达与血管生成及肿瘤转移的关系

目的探讨EphB4受体与其配体EphrinB2在胃癌组织中的表达及其与血管生成和肿瘤转移的关系。方法应用免疫组织化学(免疫组化)SABC法检测93例胃癌和30例癌旁组织中EphB4受体与其配体EphrinB2的表达及间质微血管密度(MVD)。结果胃癌组织中肿瘤细胞、间质新生血管的EphB4与EphrinB2高度表达。癌组织中EphB4与EphrinB2的阳性表达(49.5%和50.5%)明显高于癌旁组织(20.0%和23.3%,P=0.005和P=0.009);EphB4与EphrinB2阳性表达组的平均MVD值(56.21±15.3和62.41±16.9)明显高于EphB4与EphrinB2阴性表达组(32.22±12.9和34.12±14.7),两组比较,分别为P=0.018和P=0.004。此外,EphB4与EphrinB2表达程度还与胃癌的转移和癌浸润程度密切相关。结论EphB4与EphrinB2可以促进肿瘤间质的血管生成,加速肿瘤浸润和转移。     

凋亡相关基因在骨肉瘤细胞中的表达及其临床意义

目的 探讨凋亡相关基因在骨肉瘤细胞中的表达及其临床意义.方法 采用3个公认的骨肉瘤细胞株及1个成骨细胞株,提取总RNA,合成生物素标记的cRNA与Affymetrix(R)GeneChip(R) U133A芯片杂交,筛选差异表达的凋亡相关基因.利用MATLAB软件进行细胞凋亡通路的KEGG分析.结果 以表达差异≥2.0倍为限,在Affymetrix(R)HG-U133A芯片包含的所有基因中,3个骨肉瘤细胞株与成骨细胞株比较,一共筛选出7个差异表达的凋亡相关基因:3个上调基因(IKBKB、IL1R1和FAS),4个下调基因(TP53、PPP3CB、PPP3CC和APAF1).利用MATLAB软件将7个差异表达的凋亡相关基因映射到KEGG的凋亡通路上.结论 骨肉瘤足细胞增殖和细胞凋亡间平衡失调的结果,通过调控细胞凋亡相关基因的表达可以诱导肿瘤细胞凋亡,达到抗肿瘤目的 .     

结直肠癌转移相关基因表达及其临床病理意义

目的评价P53、血管内皮生长因子(VEGF)、上皮钙黏附蛋白(E-CD)、α-连环蛋白(α-CA)及β-连环蛋白(β-CA)在结直肠癌转移潜能和预后判断中的价值。方法应用免疫组织化学ABC法检测76例结直肠癌组织中P53、VEGF、E-CD、α-CA及β-CA的表达。结果(1)结直肠癌组织中P53表达阳性率为47.4%,伴有肝转移的结直肠癌P53阳性表达率明显高于无肝转移者(P<0.05);VEGF表达阳性率为57.9%,VEGF表达阳性的结直肠癌易发生淋巴结转移和肝转移(P<0.01);E-CD、α-CA和β-CA表达在结直肠癌组织中均明显减弱,且与结直肠癌的分化程度密切相关(P<0.01);E-CD表达与结直肠癌转移无明显的相关关系(P>0.05);α-CA、β-CA表达减弱与结直肠癌淋巴结转移显著相关(P<0.01),但与肝转移发生无相关(P>0.05)。(2)P53与VEGF表达显著相关(P<0.05)。(3)Cox模型多因素分析表明,P53和VEGF表达阴性的结直肠癌患者预后较好,而α-CA和β-CA表达减弱者预后较差(P<0.01)。结论结合分析P53、VEGF、α-CA及β-CA的表达有助于对结直肠癌转移方式、转移潜能及预后的判断。     

Mycophenolate mofetil inhibits tumor growth and angiogenesis in vitro but has variable antitumor effects in vivo, possibly related to bioavailability

BACKGROUND: Identifying immunosuppressive agents with antitumor effects could help address the problem of posttransplant malignancy. Here we tested for potential inhibitory effects of mycophenolate mofetil (MMF) on tumors in vitro and in vivo. METHODS: Mouse CT26 colon adenocarcinoma, B16 melanoma, and human TMK1 gastric adenocarcinoma cells were tested for in vitro growth in the presence of MMF. In vitro angiogenesis was tested with a rat aortic-ring assay. Tumor cells were implanted into dorsal skinfold chambers (DSFC) to assess in vivo angiogenesis. Subcutaneous tumor growth was determined in mice receiving MMF. RESULTS: MMF caused a dose-dependent reduction in tumor cell numbers in vitro, starting between 0.1 to 1 microM. Vessel sprouting from aortic rings was markedly blocked by similar concentrations of MMF. In vivo, however, DSFC results showed a marginal reduction in CT26 tumor angiogenesis with MMF doses of 40 or 80 mg/kg/day, although MMF did inhibit TMK1 vascularity. Moreover, 80 mg/kg/day MMF did not reduce subcutaneous CT26 tumor volumes, but did slightly inhibit B16 and TMK1 expansion. Interestingly, the mycophenolic acid (MPA) blood level 2 hr after 80 mg/kg/day MMF bolus dosing was near 14 mg/L, but decreased dramatically thereafter, suggesting a drug availability issue. Indeed, intermittent 2-hr MMF pulses in tumor-cell cultures substantially reduced the antiproliferative effect of MPA. CONCLUSION: MMF strongly inhibits tumor cell growth and angiogenesis in vitro, but only marginally inhibits tumors in vivo. These contrasting results may relate to drug availability, where intermittent exposure of tumor cells to immunosuppressive doses of MMF substantially reduce its potential antitumor effects.     

脂蛋白基因在早期骨关节炎软骨下骨的表达

目的：研究早期骨关节炎软骨下骨脂蛋白相关的基因表达改变情况。方法：大鼠分为实验组（15只）和对照组（15只）.实验组切除右膝内侧半月板及内侧副韧带,对照组仅切开关节囊。于术后1、2、4周取右膝关节标本,采用全基因表达谱芯片技术研究软骨下骨全基因表达,利用差异基因分析的方法分析脂蛋白相关的基因表达改变情况。结果：软骨下骨Apoa5表达于建模术后1周上调,术后2周下调;Apoc2表达于术后2周上调;Apol3表达于术后1周上调,术后4周下调;Lrp1于术后1、2周下调;Lrp5于术后2周下调;Gpihbp1、Lpl、Tfpi、Vldlr表达于术后1周均上调;Lrpap1、RGD1309808于术后4周均下调。结论：脂蛋白相关基因的表达改变在早期膝关节骨关节炎的软骨下骨变化中可能起了重要作用。     

METH1在酵母双杂交系统中的表达及对报告基因激活作用的检测

目的通过观察血管生成抑制因子METH1的cDNA片段在酵母双杂交中的表达及检测其对报告基因有无激活作用，为进一步明确METHI抑制增生性瘢痕的分子机制奠定基础。方法采用酵母双杂交Ga14系统3，经PCR扩增子METH1的cDNA片段，分别克隆入pUC19质粒，经测序正确后。再分别亚克隆入酵母双杂交诱饵载体pGBKT7中。将重组质粒导入酵母菌AH109，检测其表达产物在酵母细胞中对报告基因的激活作用。结果成功获得METH1的cDNA片段，该片段所表达的蛋白对酵母菌AH109无毒性，且对报告基因无激活作用。结论血管生成抑制因子METH1蛋白活性区在酵母双杂交系统中的表达产物。可作为诱饵蛋白进行相互作用蛋白的筛选研究。     

METH1在酵母双杂交系统中的表达及对报告基因激活作用的检测

目的通过观察血管生成抑制因子METH1的cDNA片段在酵母双杂交中的表达及检测其对报告基因有无激活作用,为进一步明确METH1抑制增生性瘢痕的分子机制奠定基础。方法采用酵母双杂交Gal4系统3,经PCR扩增子METH1的cDNA片段,分别克隆入pUC19质粒,经测序正确后,再分别亚克隆入酵母双杂交诱饵载体pGBKT7中。将重组质粒导入酵母菌AH109,检测其表达产物在酵母细胞中对报告基因的激活作用。结果成功获得METH1的cDNA片段,该片段所表达的蛋白对酵母菌AH109无毒性,且对报告基因无激活作用。结论血管生成抑制因子METH1蛋白活性区在酵母双杂交系统中的表达产物,可作为诱饵蛋白进行相互作用蛋白的筛选研究。     

Urodynamic findings in patients with nocturia and their associations with patient characteristics

IntroductionThis study identified associations between lower urinary tract pathology confirmed on urodynamic testing, baseline characteristics, and symptoms for adults with nocturia. Nocturia frequency was examined for predictors.MethodsThis retrospective study from 2012–2019 analyzed adult patients with nocturia (waking to void ≥2x/night) referred for urodynamic testing (UDS). Data on baseline characteristics, symptoms, UDS parameters, and lower urinary tract pathology were recorded. Males and females were analyzed separately, and univariable analyses were conducted, stratified by lower urinary tract pathology. Multivariable regression models were fit. Nocturia frequency was analyzed for associations with clinical parameters.ResultsAltogether, 372 patients were included (159 men and 213 women). More men had detrusor overactivity (DO) (p<0.001) and bladder outlet obstruction (BOO) (p<0.001). DO was associated with storage symptoms (odds ratio [OR] 5.19, p<0.001), in addition to older age (p=0.009) and being male (p<0.001). Detrusor under-activity (DU) was associated with voiding symptoms (OR 1.92, p=0.004), older age (p<0.001), and being female (p=0.018). BOO was associated voiding symptoms (OR 2.09, p=0.023), younger age (p=0.018), and being male (p<0.001). The quantity of lower urinary tract symptoms was associated with DU and DO. Nocturia frequency was not associated with baseline variables or underlying pathologies. A substantial number of patients were diagnosed with DU alone (n=69, 18.7%) or associated with other diagnoses (n=108, 29.3%).ConclusionsCareful assessment of risk factors and symptoms may help identify underlying lower urinary tract pathology for adults with nocturia. DU is found in a significant proportion of patients with nocturia, a previously under-reported result.

KEY MESSAGES

• In adults presenting with nocturia for urodynamic testing, storage and voiding lower urinary tract symptoms (LUTS) may help differentiate underlying lower urinary tract pathology.
• The quantity of LUTS may increase the likelihood of being diagnosed with certain lower urinary tract pathologies.
• A substantial proportion of adults presenting with nocturia have underlying detrusor underactivity, a known underdiagnosed condition.