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首页> 外文期刊>Urology >Integrity of prostatic tissue for molecular analysis after robotic-assisted laparoscopic and open prostatectomy.
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Integrity of prostatic tissue for molecular analysis after robotic-assisted laparoscopic and open prostatectomy.

机译:机器人辅助腹腔镜和开放式前列腺切除术后用于分子分析的前列腺组织的完整性。

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OBJECTIVES: The warm ischemia time of tissue before fixation for pathologic analysis has been linked to changes in cell morphology and nucleic acid and protein integrity. Robotic-assisted laparoscopic prostatectomy (RALP) results in longer warm ischemia times than open radical retropubic prostatectomy. To assess the effect of longer ischemia times on biomolecular integrity, we analyzed DNA, RNA and protein collected from robotic and open prostatectomy specimens. METHODS: We examined 22 consecutive RALP (n = 11) and open radical retropubic prostatectomy (n = 11) specimens after hematoxylin-eosin staining by light microscopy. To assess protein integrity, immunohistochemical staining for p63, E-cadherin, and AE1/AE3 was performed. DNA was assessed by gel analysis. An RNA integrity score was determined by microfluidic capillary electrophoresis and calculated according to the electropherogram findings and simulated gel view. Finally, epithelial cells were cultured on collagen-coated plates. RESULTS: No differences in clinicopathologic characteristics were found between the two groups, with the exception of a significantly longer warm ischemia time during RALP (82 +/- 23 minutes) compared with open radical retropubic prostatectomy (23 +/- 2 minutes; P <0.001). Tissue integrity was suitable for the assessment of pathologic grade and stage for all samples. Protein and DNA analyses demonstrated no evidence of degradation in any samples. No significant differences in the RNA integrity scores were demonstrated between the surgical approaches. Prostate epithelial cells were cultured successfully in 66% of the RALP specimens. CONCLUSIONS: RALP, although it involves additional exposure to warm ischemia, does not significantly affect the histopathologic characteristics or biomolecular integrity of the specimen. Provided a rapid response occurs for tissue banking after specimen removal, molecular research studies using prostatic tissue harvested by way of RALP appear feasible.
机译:目的:在进行病理分析固定之前,组织的温暖缺血时间与细胞形态以及核酸和蛋白质完整性的变化有关。与开放式根治性耻骨后前列腺切除术相比,机器人腹腔镜前列腺切除术(RALP)导致的缺血时间更长。为了评估更长的缺血时间对生物分子完整性的影响,我们分析了从机器人和开放式前列腺切除术标本中收集的DNA,RNA和蛋白质。方法:我们通过光学显微镜检查了苏木精-曙红染色后的22个连续RALP(n = 11)和开放根治性耻骨后前列腺切除术(n = 11)标本。为了评估蛋白质的完整性,对p63,E-钙黏着蛋白和AE1 / AE3进行了免疫组织化学染色。通过凝胶分析评估DNA。 RNA完整性评分通过微流体毛细管电泳确定,并根据电泳图结果和模拟凝胶视图进行计算。最后,将上皮细胞培养在胶原蛋白包被的板上。结果:两组患者在临床病理特征上无差异,不同之处在于,与开放式根治性耻骨后前列腺切除术(23 +/- 2分钟)相比,RALP期间的温暖缺血时间(82 +/- 23分钟)明显更长。 0.001)。组织完整性适合评估所有样品的病理分级和分期。蛋白质和DNA分析表明,没有任何样品降解的迹象。在两种手术方法之间,RNA完整性评分无明显差异。在66%的RALP标本中成功培养了前列腺上皮细胞。结论:RALP尽管涉及额外的热缺血,但不会显着影响标本的组织病理学特征或生物分子完整性。如果在去除标本后迅速对组织库进行反应,那么使用通过RALP收集的前列腺组织进行的分子研究似乎是可行的。

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