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大豆蛋白在甲状腺细针穿刺细胞蜡块制备中的应用OA

Application of soybean protein in the preparation of cell blocks from thyroid fine-needle aspiration samples

中文摘要英文摘要

目的:甲状腺细针穿刺抽吸术(fine-needle aspiration,FNA)是评估甲状腺结节性质的首选方法,但传统涂片存在细胞量有限、组织结构信息丢失、难以满足后续免疫组织化学及分子检测需求等问题.细胞蜡块技术可弥补上述不足,但现有制备方法在处理微量细胞样本时存在细胞丢失率高、操作烦琐、外源污染风险高等问题.本研究旨在通过探讨大豆蛋白联合滤纸转移法制备甲状腺FNA细胞蜡块的成功率及其临床应用价值,建立一种操作简便、成本低廉、能有效富集微量细胞的细胞蜡块制备新方法,为基层医疗机构提供可行的技术方案.方法:回顾性收集2025年1月至10月于应城市人民医院超声影像科行超声引导下甲状腺FNA检查的80例患者样本,其中男性23例(28.8%),女性57例(71.2%),年龄范围为26~88岁,年龄为(53.9±11.4)岁.所有样本同时制备传统涂片和细胞蜡块.细胞蜡块采用大豆蛋白联合滤纸转移法制备:将穿刺物立即冲洗固定,离心富集细胞后,加入大豆蛋白载体液及95%乙醇,混匀后置入圆形滤纸,经离心使细胞沉淀被滤纸有效截留并完整承载于管底,随后取出沉淀进行脱水、包埋、切片.评估指标包括细胞蜡块制备成功率、苏木精-伊红(hematoxylin and eosin,HE)染色质量及免疫组织化学染色效果.由2位高年资病理医师采用双盲法独立完成Bethesda诊断系统诊断分类,比较传统涂片与细胞蜡块诊断分类的差异.统计学方法采用Cohen's Kappa检验评估2种方法诊断分类的一致性,采用McNemar检验(连续性校正)比较2种方法在不确定诊断(Bethesda Ⅲ类+Ⅴ类)分类上的差异,计算卡方(χ2)值、P值、配对OR及其95%CI,检验水准α=0.05.结果:细胞蜡块制备成功率为96.25%(77/80),3例失败样本涂片细胞量均低于1 000个;HE染色显示细胞形态保存完好,染色质结构清晰,细胞核-细胞质对比鲜明;大豆蛋白背景呈浅红色均匀分布,对目标细胞无遮蔽效应;偶见植物细胞碎片易于鉴别,未干扰诊断;免疫组织化学染色定位准确、背景干净,细胞角蛋白19(cytokeratin 19,CK19)、甲状腺转录因子-1(thyroid transcription factor-1,TTF-1)、甲状腺过氧化物酶(thyroid peroxidase,TPO)等抗体表达模式与对应组织蜡块一致;2种方法诊断分类的总体一致性为中等(Kappa=0.532,P<0.001),完全一致病例53例,总体符合率为66.25%(53/80);细胞蜡块使不确定诊断(Ⅲ类+Ⅴ类)病例数由涂片的30例降至14例,不确定诊断率降低53.33%(16/30).其中,62.50%(10/16)的涂片Ⅴ类(可疑恶性)病例在蜡块中明确升级为Ⅵ类(恶性).McNemar检验显示差异具有统计学意义(χ2=11.250,P<0.001),配对OR=9.000(95%CI 2.160~37.480).结论:大豆蛋白联合滤纸转移法制备甲状腺FNA细胞蜡块成功率高,能有效富集微量细胞,细胞形态与抗原性保存较好,HE染色及免疫组织化学染色效果满意.该方法可显著降低Bethesda诊断系统不确定诊断率,提升甲状腺FNA诊断明确性,为后续免疫组织化学及分子检测提供了可靠平台.

Objective:Fine-needle aspiration(FNA)is the preferred method for evaluating the nature of thyroid nodules.However,conventional smears have limitations,including limited cellularity,loss of architectural information,and inability to meet the requirements for subsequent immunohistochemical and molecular testing.The cell block technique can compensate for these shortcomings,but existing preparation methods for scant cellular samples are associated with high cell loss,procedural complexity,and increased risk of exogenous contamination.This study aims to evaluate the success rate and clinical application value of a soybean protein-assisted filter paper transfer method for preparing thyroid FNA cell blocks,and to establish a simple,low-cost method capable of effectively enriching scant cellular material,thereby providing a feasible technical solution for primary healthcare institutions. Methods:A total of 80 patients who underwent ultrasound-guided thyroid FNA at the Department of Ultrasound,Yingcheng People's Hospital,from January to October 2025 were retrospectively included.Among them,23(28.8%)were male and 57(71.2%)were female,with an age range of 26 to 88 years and a mean age of 53.9±11.4 years.For each case,both conventional smears and cell blocks were prepared.The cell blocks were produced using a soybean protein-assisted filter paper transfer method:Aspirates were immediately rinsed and fixed,centrifuged to concentrate cells,followed by the addition of soybean protein carrier solution and 95%ethanol.After thorough mixing,the suspension was transferred onto circular filter paper and centrifuged,allowing cell sediment to be effectively retained and completely carried at the bottom of the tube.The sediment was then collected for dehydration,embedding,and sectioning.Evaluation indicators included the success rate of cell block preparation,hematoxylin and eosin(HE)staining quality,and immunohistochemical staining performance.Two senior pathologists independently performed diagnostic classification according to the Bethesda diagnostic system in a double-blind manner.Differences between conventional smears and cell blocks were compared.Cohen's Kappa test was used to assess diagnostic concordance,and the McNemar test(with continuity correction)was applied to compare differences in indeterminate diagnoses(Bethesda categories Ⅲ and Ⅴ),with calculation of χ2 values,P values,paired odds ratios(OR),and 95%confidence intervals(CI).The significance level was set at α=0.05. Results:The success rate of cell block preparation was 96.25%(77/80).In the three failed cases,smear cellularity was<1 000 cells.HE staining showed well-preserved cellular morphology,clear chromatin structure,and distinct nuclear-to-cytoplasmic contrast.The soybean protein background appeared as a uniformly distributed pale red matrix without obscuring target cells.Occasional plant cell debris were easily identifiable and did not interfere with diagnosis.Immunohistochemical staining showed accurate localization and clean background;the expression patterns of cytokeratin 19(CK19),thyroid transcription factor-1(TTF-1),and thyroid peroxidase(TPO)were consistent with those observed in corresponding tissue blocks.The overall diagnostic concordance between the 2 methods was moderate(Kappa=0.532,P<0.001),with complete agreement in 53 cases and an overall concordance rate of 66.25%(53/80).The use of cell blocks reduced the number of indeterminate diagnoses(categories Ⅲ and Ⅴ)from 30 cases in smears to 14 cases,representing a 53.33%reduction(16/30).Notably,62.50%(10/16)of smear-based categoryⅤ(suspicious for malignancy)cases were upgraded to category Ⅵ(malignant)in cell blocks.McNemar test showed a statistically significant difference(χ2=11.250,P<0.001),with a paired OR of 9.000(95%CI 2.160 to 37.480). Conclusion:The soybean protein-assisted filter paper transfer method for preparing thyroid FNA cell blocks demonstrates a high success rate and effectively enriches scant cellular material,with good preservation of cellular morphology and antigenicity.Both HE staining and immunohistochemical staining show satisfactory performance.This method significantly reduces the rate of indeterminate diagnoses in the Bethesda diagnostic system and improves diagnostic certainty in thyroid FNA,providing a reliable platform for subsequent immunohistochemical and molecular testing.

张贝;夏磊;彭琪彦;张雪莲;岳慧萍

应城市人民医院病理科,孝感 432400应城市人民医院超声影像科,孝感 432400应城市人民医院病理科,孝感 432400应城市人民医院病理科,孝感 432400应城市人民医院病理科,孝感 432400

医药卫生

大豆蛋白细胞蜡块甲状腺结节细针穿刺细胞病理

soybean proteincell blockthyroid nodulefine-needle aspirationcytopathology

《临床与病理杂志》 2026 (2)

252-262,11

孝感市自然科学计划(XGKJ2024010033).This work was supported by the Xiaogan City Natural Science Program,China(XGKJ2024010033).

10.11817/j.issn.2095-6959.2026.251011

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