p16/Ki-67手工染色与全自动免疫组化仪染色效果对比

<正>p16蛋白是细胞周期蛋白依赖性激酶抑制剂,p16在细胞周期的阻滞期具有抑制细胞增殖的功能^[1]。Ki-67表示细胞周期进展及增殖活跃^[2]。两者同时表达在一个细胞周期则提示细胞周期失调。正常生理状态下,p16和Ki-67的表达相互制约,不会同时表达于一个细胞中。若p16/Ki-67免疫细胞化学双染检测阳性,提示子宫颈上皮内病变2级（cervical intraepithelial neoplasias 2,CIN2）及以上病变^[3-6]。     

探讨外院会诊玻片行全自动免疫组化仪染色异常的优化方法

正免疫组化技术可以检测细胞来源、辨认细胞产物及了解分化程度等,为病理诊断和鉴别诊断提供客观依据,并为疾病治疗提供指导。在精准医疗时代,全自动免疫组化仪以其自动化程度高、操作简便、染色流程标准化等特点,在病理科广泛应用~([1-2])。但全自动免疫组化仪染色也会遇到组织玻片非特异染色、染色偏弱或无法着色等问题~([3]),其中以外院送检的会诊组织在罗氏Ventana免疫组化染色时最为常见。     

全自动免疫组化仪与手工操作应用体会

免疫组化技术作为病理诊断的重要辅助手段，在临床病理鉴别诊断、肿瘤评估及耐药检测等方面发挥不可或缺的作用，大大提高病理诊断水平。近年随着社会总人口以及癌症患者数量增长，病理科的工作量相应增大，传统的手工操作已不能满足工作需要，因此各种型号的自动免疫组化仪应运而生。本科室于2014年3月开始使用全自动免疫组化染色仪，经过一段时间的实践摸索，现将其应用体会总结如下。     

全自动免疫组化染色仪的应用体会

目前国内免疫组化多采用手工操作,虽有不少单位使用了全自动免疫组化染色仪,但是使用大容量全自动免疫组化染色仪的报道少见.我们通过摸索实践,在以往手工操作的基础上,充分利用全自动免疫组化染色仪流程的优点,密切注意染色过程中所出现的问题并加以改进,加强对仪器的保养维护,确保免疫组化染色质量的稳定性.现将体会介绍如下.     

优化全自动免疫组化仪切片贴片位置

<正>随着免疫组化染色标准化、规范化的推行，全自动免疫组化仪以其操作简单、重复性高、染色流程标准化等优点被广泛应用。但免疫组化的染色步骤繁多，稍有失误就会影响染色结果，进而影响病理诊断的准确性^[1-2]。设立对照是控制染色质量的重要措施之一，可以根据对照结果判断免疫组化染色过程中操作步骤的可行性、使用试剂的可靠性^[3]。日常工作中，因组织贴片区域不恰当，尤其是阳性对照贴片位置偏离全自动免疫组化染色区域，不可避免的会出现一些染色假象，     

自动免疫组化仪的使用体会

免疫组化技术在病理诊断中的应用日益广泛,并已成为常规病理诊断中不可缺少的重要手段.良好的免疫组化制片是正确判断染色结果的前提和基础.但由于免疫组化染色有很多步骤,每一步都有可能影响染色结果[1],对相同的抗体会因为不同的实验室操作程序、操作人员素质的高低而导致结果有所差异[2],因此正确使用自动免疫组化仪,将免疫组化的操作标准化,达到免疫组化的质量控制要求,是病理技术质量控制的必然趋势.本文通过摸索实践,总结出全自动免疫组化染色仪的使用体会,介绍如下.     

自动免疫组化染色仪染色的质量控制

影响免疫组化实验结果的因素很多,如检测系统灵敏度、抗原修复方法不同等。因此,免疫组化标准化和质量控制势在必行。现将我们的工作总结如下。     

提高全自动免疫组化仪制片质量的相关细节

为确保免疫组化染色结果的重复性和真实性,现代病理工作者已逐渐将目光转向高品质的全自动免疫组化仪,但自动化染色也可能产生一些假象.我们在使用美国Bench Mark XT全自动免疫组化仪后总结了针对提高制片质量的几点建议,与大家共同探讨. 1 标本固定、处理及蜡块选择的相关细节 免疫组化染色过程中抗原性保存完整、不弥散或丢失是实验成功的重要保证.在一张组织或细胞已坏死、抗原已丢失的材料上,再好的技术和抗体也难以成功染色[1].组织处理之前必须保证组织充分固定,如果固定不适当、不完全,可能会改变组织的染色特性[2].     

Infant pleuropulmonary blastoma: report of a rare case and review of literature

In infants, pleuropulmonary blastoma is a rare but aggressive tumor. The typical histopathological presentation includes the aggregation of malignant primitive small cells, usually observed in sheets. So as to provide proper and timely treatment, the differential diagnosis includes pulmonary blastoma, sarcomatoid mesothelioma, fetal rhabdomyoma, synovial sarcoma, and primitive neuroectodermal tumor. Herein, we will present one male pediatric patient with pleuropulmonary blastoma. The patient was a 4-month-old male infant, who had a prolonged cough and dyspnea for 4 months that was complicated by cyanosis for 3 days. A physical examination revealed a solid mass in the right lung that was sized 9.0 × 6.0 × 4.0 cm and had a grayish-white cross section. The boundary between the mass and lung tissue was clear; the mass already occupied a great portion of the lung. A microscopic examination suggested that the tumor was composed of round or orbicular-ovate primitive fetal cells. The cells were medium sized, having little cytoplasm, but had a clearly visualized nucleolus and active karyokinesis. The tumor mass was biphasic, namely, fasciculated sarcoma (composed of spindle-shaped cells and short spindle-shaped cells) and malignant fibrous histiocytoma containing well-differentiated cartilage islands or cartilaginous nodes. Immunohistochemistry was performed for further detection: vimentin (+), S-100 protein (+), CK (AE1/AE3), EMA and TTF-1 in residual epithelial components (+), NSE (focal +), SMA (mesenchymal cells, focal +), CD99 (weak +), Bcl-2 (weak +), desmin (-), myoglobin (-), calretinin (-), calponin (-), FLI (-), MyoD-1 (-), and CD34 (-). Pleuropulmonary blastoma is extremely rare but highly aggressive neoplasm in children. Its typical histopathological presentation is the aggregation of primitive malignant small cells. Combining imaging and histopathological examinations and clinical data should help in determining the diagnosis of pleural pulmonary blastoma.     

QTD自动测量与人工测量对比研究

目的：研究QTD自动测量技术及测量结果与人工测量结果的对比。方法：取得卧、立位的心电信号，CM5导联，采样率300Hz,ADC12位，放大器放大倍数1500倍，用最大值法和阈值法自动测量QTD，人工目测法测量QTD。每例至少测三个心动周期，用相关分析技术检验三者的相关性。结果：三种测量方法中，以最大值法的可靠性最高，重复性最好，对噪声的敏感性小，且不受阀值设置的影响，三者间的两两相关系数为：R（自动峰值测量与人工测量）＝0．998353，R（自动峰值测量与自动始末点测量）＝0．997849，R（自动始末点测量与人工测量）＝0．997532，结论：可用最大值法的自动测量QTD结果，代替起点一终点法测量QTD的结果。讨论：（1）若采用最大值法的自动测量结果，作为肌缺血，心肌梗塞等心血管疾病的预测和评价指标，必须重新正常值；（2）当前发表的文章，大多为人工测量结果：（3）当前有一些计算自动测量QTD的设备进入临床，但临床多反映可靠性差，可能皆为测量方法可靠性差所致。     

Somatostatin receptor immunohistochemistry in neuroendocrine tumors: comparison between manual and automated evaluation

Background: Manual evaluation of somatostatin receptor (SSTR) immunohistochemistry (IHC) is a time-consuming and cost-intensive procedure. Aim of the study was to compare manual evaluation of SSTR subtype IHC to an automated software-based analysis, and to in-vivo imaging by SSTR-based PET/CT. Methods: We examined 25 gastroenteropancreatic neuroendocrine tumor (GEP-NET) patients and correlated their in-vivo SSTR-PET/CT data (determined by the standardized uptake values SUVmax,-mean) with the corresponding ex-vivo IHC data of SSTR subtype (1, 2A, 4, 5) expression. Exactly the same lesions were imaged by PET/CT, resected and analyzed by IHC in each patient. After manual evaluation, the IHC slides were digitized and automatically evaluated for SSTR expression by Definiens XD software. A virtual IHC score “BB1” was created for comparing the manual and automated analysis of SSTR expression. Results: BB1 showed a significant correlation with the corresponding conventionally determined Her2/neu score of the SSTR-subtypes 2A (r_s: 0.57), 4 (r_s: 0.44) and 5 (r_s: 0.43). BB1 of SSTR2A also significantly correlated with the SUVmax (r_s: 0.41) and the SUVmean (r_s: 0.50). Likewise, a significant correlation was seen between the conventionally evaluated SSTR2A status and the SUVmax (r_s: 0.42) and SUVmean (r_s: 0.62).Conclusion: Our data demonstrate that the evaluation of the SSTR status by automated analysis (BB1 score), using digitized histopathology slides (“virtual microscopy”), corresponds well with the SSTR2A, 4 and 5 expression as determined by conventional manual histopathology. The BB1 score also exhibited a significant association to the SSTR-PET/CT data in accordance with the high affinity profile of the SSTR analogues used for imaging.     

1例气球细胞痣临床及病理分析

目的：学习气球细胞痣的临床病理诊断及鉴别诊断。方法：对1例气球细胞痣进行临床资料、病理形态及免疫组织化学观察,并结合文献对其诊断及鉴别诊断进行学习。结果：前胸壁皮肤先天性色素痣,近1年来增大,其组织学上表现为混合痣伴多量细胞胞浆丰富,呈气球样。免疫组织化学标记气球样细胞Melan A、S100、HMB45阳性,CD68阴性。结论：气球细胞痣是较为罕见的一种黑素细胞痣,根据其临床及组织学特征,结合免疫组织化学方法,有助于其诊断及鉴别诊断。     

The value of electronmicroscopy and immunohistochemistry in the diagnosis of soft tissue sarcomas: a study of 200 cases

Two hundred soft tissue sarcomas, accrued consecutively over a 4-year period, were examined by light and electronmicroscopy and by routine immunohistochemistry. The commonest tumour type was malignant fibrous histiocytoma. Fibrosarcoma, composed only of fibroblasts, was diagnosed in only one case; three others, composed also of myofibroblasts, could be regarded as fibrosarcomas or myofibrosarcomas. Immunohistochemistry was of most value in the diagnosis of rhabdomyosarcoma, leiomyosarcoma, malignant peripheral nerve sheath tumour of Schwann cell type and epithelioid sarcoma. Electronmicroscopy was of most use for the diagnosis of malignant peripheral nerve sheath tumour of perineurial cell type and marker-negative monophasic synovial sarcoma, and for confirming malignant fibrous histiocytoma. Fifteen of 19 marker-negative spindle cell tumours (79%) were diagnosable by electronmicroscopy. A combination of techniques resulted in a specific diagnosis in 193 cases (96.5%). The routine use of electronmicroscopy in sarcoma diagnosis can improve accuracy of diagnosis, establish the true frequency of marker-positivity for each ultrastructurally confirmed tumour type and minimise the number of unclassifiable cases.     

Wolffian附件肿瘤的临床病理观察

目的 观察Wolffian附件肿瘤的临床病理特点,探讨其病理诊断与鉴别诊断。方法 对3例Wolffian附件肿瘤进行HE及免疫组化染色（EnVision法）,并进行病理观察。结果 3例Wolffian附件肿瘤均为单侧,位于阔韧带或输卵管系膜。镜下肿瘤细胞呈弥漫实体状分布,可见成片的梭形或多边形细胞及排列紧密的管状结构。管腔内衬立方或柱状上皮,细胞无明显异型,核分裂象少见。管周有PAS阳性的基膜物质。免疫组化示肿瘤细胞vimentin、AE1／AE3、α-inhibin、calretinin和WT1均呈阳性,CD99呈灶性阳性,EMA和CK7呈阴性。结论 Wolffian附件肿瘤是具有独特发病部位和组织病理学特点的妇科肿瘤,需要和一系列其他妇科肿瘤鉴别。     

The comparative roles of electron microscopy and immunohistochemistry in the diagnosis of soft tissue tumours

Electron microscopy has contributed to the diagnosis of soft tissue tumours for four decades, and immunohistochemistry for two. Because of its relative ease of use and interpretation, the latter technique has become extensively and routinely applied to identify lines of differentiation in benign soft tissue tumours and in sarcomas. The use of electron microscopy has declined but retains a role because few antibodies are wholly specific or fully sensitive, some tumours are polyphenotypic or divergent in differentiation, and others have no specific antigens. Immunohistochemistry is superior in diagnosis of smooth muscle tumours, small round cell tumours, sarcomas with epithelioid morphology, and most synovial sarcomas. Electron microscopy is of particular value for peripheral nerve sheath tumours, marker-negative synovial sarcomas, pleomorphic sarcomas and mesotheliomas. As with all adjunctive techniques, immunohistochemistry and electron microscopy should be used in a complementary fashion according to the nature of the diagnostic problem.     

29例上皮样胃肠间质瘤的临床病理特征

目的:探讨29例上皮样胃肠间质瘤(gastrointestinal stromal tumors,GIST)的组织学形态和免疫组织化学特点以及诊断和鉴别诊断,对临床正确诊断治疗和判断预后具有十分重要的临床意义.方法:回顾性分析安阳市肿瘤医院病理科2009年月3月至2016年8月321例完整切除GIST标本,经筛查并重新阅片诊断上皮样GIST 29例.结果:29例上皮样GIST,发生胃部15例,小肠2例,肠系膜3例,直肠4例,腹腔3例,腹膜后1例,盆腔1例.恶性GIST 25例,良性4例.瘤细胞丰富,胞质嗜酸或透明,部分瘤细胞核质比高,核大小不等具有多形性,核分裂较多,可伴有多灶凝固性坏死,间质多数伴有黏液样变性.组织结构形成器官样、片状、巢状及腺泡状等.免疫组织化学CD34,DOG-1在上皮样GIST均弥漫阳性(阳性率100%),CD117阳性率(86%).结论:上皮样GIST发生部位广泛,形态多变,易误诊其他上皮样分化的肿瘤;免疫组织化学CD34,DOG-1,CD117在上皮样GIST诊断及鉴别诊断中具有重要价值;如肿物较大、细胞丰富、核分裂多、间质黏液样变性;绝大多数要考虑恶性GIST.     

Ki-67/MIB-1 immunostaining in a cohort of human gliomas

Histopathological malignancy grading of human gliomas is limited by subjective interpretation of the morphological criteria. Assessment of mitotic activity is a cornerstone of grading these tumours, but mitotic figures can be hard to identify in haematoxylin-eosin stained sections. Thus, determining proliferative activity by means of Ki-67/MIB-1 immunostaining has become a useful supplement. However, this method has drawbacks, so continuous testing and evaluation are required for optimization and standardization. The aim of this study was to analyse and evaluate the Ki-67/MIB-1 proliferative indices (PIs) in a series of gliomas. We found that Ki-67/MIB-1 PIs correlated well with histological malignancy grade in all glioma subtypes, but a considerable overlap of PIs was observed between the malignancy groups. Consequently, Ki-67/MIB-1 immunostaining alone is not sufficient to adequately determine the malignancy grade. Therefore, future work is necessary to clarify the role of this immunostaining in the histopathological diagnosis of human gliomas.