低渗破红法在非小细胞肺癌患者血性胸水中的应用
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莆田学院 附属医院,莆田3511001.病理科; 2.医务部

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Application of Low-Osmosis to Eliminated Erythrocyte in NSCLC Patients
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1.Department of Pathology, The Affiliated Hospital of Putian University, Putian 351100, China; 2.Department of Medical Administration, The Affiliated Hospital of Putian University, Putian 351100, China

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    摘要:

    目的探讨低渗破红法对非小细胞肺癌(NSCLC)患者血性胸水中肿瘤细胞形态的影响及其在分子病理检测前评估中的应用。方法收集2018年3月-2019年3月送检的经病理诊断为NSCLC的血性胸水样本24例,采用低渗破红法处理并制备成细胞块,并以50%乙醇+10%中性缓冲甲醛混合液破红和常规细胞块技术为对照,切片经H-E染色进行肿瘤细胞形态分析。此外,分别切取细胞蜡卷提取核酸,进行分子病理检测前的评估,并对低渗破红法处理制备的细胞块所提取的核酸进行肺癌靶向治疗相关基因检测。结果低渗破红法处理制备的细胞块与常规细胞块技术制备的细胞块相比较,切片中红细胞明显减少、肿瘤细胞丰度高,染色背景清晰,轮廓结构清楚,但与50%乙醇+10%中性缓冲甲醛混合液破红法相比较,差别无统计学意义(P>0.05); 细胞块核酸合格率与常规细胞块技术法相比较差别显著(P<0.05),与50%乙醇+10%中性缓冲甲醛混合液破红法相比较差别无统计学意义(P>0.05)。低渗破红法制备的细胞块行肺癌靶向治疗相关基因检测,其中13例检测出EGFR基因突变,未检测出EML4-ALK和ROS1融合基因。结论低渗破红法是处理NSCLC患者血性胸水良好的有效方法,并能够支持肺癌靶向治疗相关的基因检测。

    Abstract:

    ObjectiveTo explore the impacts of eliminated erythrocyte on morphological alterations of tumour cells in hemorrhagic effusion from NSCLC carcinoma and the pretreatment application of gene mutation detection by low-osmosis method.MethodsData of 24 cases of hemorrhagic effusion from NSCLC carcinoma patients in the Affiliated Hospital of Putian University from march 2018 to march 2019 were collected.Cytologic blocks were prepared with paraffin-embedding by low-osmosis to remove RBCs.We used mixture of 50% ethanol and 10% neutral buffer formalin(NBF)to damages RBCs and conventional way to form a paraffin-embedded cytologic block in control group.H-E and nucleic acid extraction evaluations were performed.ResultSignificantly decreased RBCs, higher number of tumor cells, clearly and more distinct cellular outlines were significantly different with low-osmosis method than the conventional method(P<0.05).There was no obvious difference compared to the mixture of 50% ethanol and 10% NBF(P>0.05).The qualification rate of nucleic acids-both DNA and RNA-by low-osmosis method was noticeably different than that of the conventional method(P<0.05).Thirteen nucleic acid mutations were detected in 24 cases screened for EGFR mutations while EML4-ALK and ROS1 gene fusion were not detected.ConclusionThe results indicate that the low-osmosis method is an efficient way to dispose the hemorrhagic effusion from NSCLC carcinoma and it is able to support the genetic testing related to target therapy of NSCLC.

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刘芳芳, 陈智伟, 祁晓育.低渗破红法在非小细胞肺癌患者血性胸水中的应用[J].福建医科大学学报自然版,2020,(4):269-273

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