• 细针穿刺标本Midkine检测在甲状腺微小乳头状癌中 不要轻易放弃。学习成长的路上,我们长路漫漫,只因学无止境。


    探讨细针穿刺标本Midkine检测在甲状腺微小乳头状癌中的临床意义。方法 选取2014年8月~2017年5月我院收治的50例甲状腺微小乳头状癌患者、30例结节性甲状腺肿患者的术中细针穿刺标本作为研究对象,观察两类病理类型免疫细胞化学技术检测Midkine的表达。结果 甲状腺微小乳头状癌细针穿刺标本Midkine的表达明显高于结节性甲状腺肿,差异有统计学意义(P0.05),具有可比性。本研究经我院医学伦理委员会审核批准,所有患者均知晓本研究情况并签署知情同意书。 1.2方法 1.2.1标本采集 用22 G针头接10 ml一次性注射器,在甲状腺结节中多点、反复负压抽吸3~5次,将抽出物直接均匀涂于3~8张多聚赖氨酸处理过的玻片上。玻片置于95%酒精中固定15 min,然后放入保湿密闭的切片盒中,置于-20℃冰箱中冷冻保存待用。 1.2.2免疫细胞化学试剂 兔抗人Midkine抗体购自北京北方生物技术研究所有限公司;兔超敏二步法免疫组化检测试剂盒(PV9001)及二氨基联苯胺显色液均购自北京中杉金桥生物技术有限公司;苏木素购自江苏凯基生物技术有限公司。 1.2.3免疫细胞化学实验方法 于冰箱中取出细胞涂片,置室温10~15 min复温。经蒸馏水洗涤后,在柠檬酸盐缓冲液pH6.0中抗原热修复20 min。3%过氧化氢室温孵育10~20 min,阻断内源性过氧化物酶,PBS缓冲液冲洗3 min×3次。滴加牛血清白蛋白封闭,室温孵育20 min。滴加100倍稀释的一抗兔抗人Midkine抗体4℃过夜,PBS缓冲液冲洗3 min×3次。置于37℃水浴锅内复温45 min。滴加反应增强液,37℃孵育20 min,PBS缓冲液冲洗3 min×3次。滴加增强酶标山羊抗兔IgG聚合物,37℃孵育20 min,PBS缓冲液冲洗3 min×3次。二氨基联苯胺显色5 min,双蒸水冲洗。苏木素复染,梯度乙醇脱水,二甲苯透明,中性树胶封片观察。用已知阳性切片作阳性对照,用PBS缓冲液代替一抗作阴性对照。 1.3评价标准 以细胞胞质内有清晰棕黄色颗粒为阳性细胞。阳性细胞占比≥10%的标本判定为阳性,反之,判定为阴性。 1.4统计学方法 采用SPSS 20.0统计学软件进行数据分析,计量资料用均数±标准差(x±s)表示,两组间比较采用t检验;计数资料用率表示,组间比较采用χ2检验,以P0.05)。无淋巴结转移甲状腺微小乳头状癌患者21例,Midkine阳性表达14例(66.7%);有淋巴结转移甲状腺微小乳头状癌患者29例,Midkine阳性表达27例(93.1%),有淋巴结转移甲状腺微小乳头状癌患者的细针穿刺标本Midkine表达阳性率高于无淋巴结转移患者,差异有统计学意义(P[3]Lim H,Devesa SS,Sosa JA,et al.Trends in thyroid cancer incidence and mortality in the United States,1974-2013[J].JAMA,2017,317(13):1338-1348. [4]Oda H,Miyauchi A,Ito Y,et al.Incidences of unfavorable events in the management of low-risk papillary microcarcinoma of the thyroid by active surveillance versus immediate surgery[J].Thyroid,2016,26(1):150-155. [万博足彩,万博体育怎么下赌注,万博体育3.85]Li D,Gao M,Li X,et al.Molecular aberrance in papillary thyroid microcarcinoma bearing high aggressiveness:identifying a “tibetan mastiff dog” from puppies[J].J Cell Biochem,2016,117(7):1491-1496. [6]Leboulleux S,Tuttle RM,Pacini F,et al.Papillary thyroid microcarcinoma:time to shift from surgery to active surveillance?[J].Lancet Diabetes Endocrinol,2016,4(11):933-942. [7]Lee J,Soh EY.Current guidelines for fine needle aspiration of thyroid nodules[J].Korean J Endocrine Surg,2015,15(1):1-5. [8]Zevallos JP,Hartman CM,Kramer JR,et al.Increased thyroid cancer incidence corresponds to increased use of thyroid ultrasound and fine-needle aspiration:a study of the Veterans Affairs health care system[J].Cancer,2015,121(5):741-746. [9]Eszlinger M,Lau L,Ghaznavi S,et al.Molecular profiling of thyroid nodule fine-needle aspiration cytology[J]万博足彩,万博体育怎么下赌注,万博体育3.8.Nat Rev Endocrinol,2017,13(7):415. [10]戎?s,吴妍,姚青,等.甲状腺细针穿刺检查2043例细胞病理学分析[J].中华病理学杂志,2016,45(6):368-371. [11]Lee YH,Baek JH,Jung SL,et al.Ultrasound-guided fine needle aspiration of thyroid nodules:a consensus statement by the Korean Society of Thyroid Radiology[J].Korean J Radiol,2015,16(2):391-401. [12]Shao H,Yu X,Wang C,et al.Midkine expression is associated with clinicopathological features and BRAF mutation in papillary thyroid cancer[J].Endocrine,2014,46(2):285-291. [13]Choi YW,Kim YH,Lee J,et al.Strong immunoexpression of midkine is associated with multiple lymph node metastases in BRAFV600E papillary thyroid carcinoma[J].Human pathology,2015,46(10):1557-1565. [14]Cabanillas ME,Mcfadden DG,Durante C.Thyroid cancer[J].The Lancet,2016,388(10061):2783-2795. [15]Chiosea S,Asa SL,Berman MA,et al.Template for reporting results of 万博足彩,万博体育怎么下赌注,万博体育3.8 biomarker testing of specimens from patients with thyroid carcinoma[J].Arch Pathol Lab Med,2016,141(4):559-563. [16]Muramatsu T.Structure and function of midkine as the basis of its pharmacological effects[J].Br J Pharmacol,2014, 171(4):814-826. [17]Jones DR.Measuring midkine:the utility of midkine as a biomarker in cancer and other diseases[J].Br J Pharmacol,2014,171(12):2925-2939. [18]Maeda S,Shinchi H,Kurahara H,et al.Clinical significance of Midkine expression in pancreatic head carcinoma[J].Br J Cancer,2007,97(3):405-408. [19]Zhao ZQ,Yang S,Lu HS.Expression of Midkine and vascular endothelial growth factor in gastric cancer and the association of high levels with poor prognosis and survival[J].Mol Med Rep,2012,5(2):415-419. [20]Jham BC,Costa NL,Silva JM,et al.Midkine expression in oral squamous cell carcinoma and leukoplakia[J].J Oral Pathol Med,2012,41(1):21-26. (收稿日期:2018-03-15 本文编辑:孟庆卿)

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