微波在免疫组织化学中的应用

本文综述微波技术在免疫组织化学中的应用两头及其进展，着重介绍了微波在组织固定、抗原恢复和免疫染色方面的应用，并对微波免疫组织化学的应用前景进行了展望 。     

Proteomic expression profiling of thyroid neoplasms

Thyroid cancer is the most common endocrine neoplasm with multiple histologic subtypes, each associated with different treatments and outcomes. Differentiating benign neoplasms such as follicular adenomas from malignant entities such as follicular carcinomas and papillary carcinoma can be challenging. To define the proteomic profile of different thyroid tumors, we screened an antibody array of 330 features against five thyroid neoplasms: follicular adenoma, follicular carcinoma, papillary carcinoma, anaplastic carcinoma, and medullary carcinoma as well as normal thyroid epithelium. Eight candidate biomarkers; c-erbB-2, Stat5a, Annexin IV, IL-11, RARα, FGF7, Caspase 9, and phospho-c-myc were identified as differentially expressed on the antibody array, and validated with immunohistochemistry on tissue microarrays, with a total of 144 samples of the same variety of thyroid neoplasms. Analysis revealed c-erbB-2, Annexin IV, and Stat5a have potential clinical utility to differentiate follicular adenoma, follicular carcinoma, and papillary carcinoma from each other. By using an antibody array as a discovery platform and a tissue microarray as a first step in validation on a large number of specimens, we have identified new markers that have potential utility in the diagnosis of thyroid neoplasms.     

Co-Expression and Co-Localization of Cartilage Glycoproteins CHI3L1 and Lubricin in Osteoarthritic Cartilage: Morphological,Immunohistochemical and Gene Expression Profiles

Osteoarthritis is the most common human arthritis characterized by degeneration of articular cartilage. Several studies reported that levels of human cartilage glycoprotein chitinase 3-like-1 (CHI3L1) are known as a potential marker for the activation of chondrocytes and the progression of Osteoarthritis (OA), whereas lubricin appears to be chondroprotective. The aim of this study was to investigate the co-expression and co-localization of CHI3L1 and lubricin in normal and osteoarthritic rat articular cartilage to correlate their modified expression to a specific grade of OA. Samples of normal and osteoarthritic rat articular cartilage were analyzed by the Kellgren–Lawrence OA severity scores, the Kraus’ modified Mankin score and the Histopathology Osteoarthritis Research Society International (OARSI) system for histomorphometric evaluations, and through CHI3L1 and lubricin gene expression, immunohistochemistry and double immuno-staining analysis. The immunoexpression and the mRNA levels of lubricin increased in normal cartilage and decreased in OA cartilage (normal vs. OA, p < 0.01). By contrast, the immunoexpression and the mRNA levels of CHI3L1 increased in OA cartilage and decreased in normal cartilage (normal vs. OA, p < 0.01). Our findings are consistent with reports suggesting that these two glycoproteins are functionally associated with the development of OA and in particular with grade 2/3 of OA, suggesting that in the future they could be helpful to stage the severity and progression of the disease.     

Immunohistochemical distribution and colocalization of regulatory peptides in the carotid body

Current investigations on the immunohistochemical occurrence and co-occurrence of biogenic polypeptides in the mammalian carotid body were reviewed and extended by our own recent findings. The family of chromogranins and related peptides in glomus cells appears to have a widespread interspecies distribution, whereas other peptides investigated occur in a species-specific pattern. Immunoreactivity to antisera against opioids, which derive from the proenkephalin sequence, appears to be present in glomus cells of the rabbit, cat, dog, and a shrew. Conversely, glomus cells of pig and guinea pig predominantly are immunoreactive to cleavage products of prodynorphin, which co-occur in some cells with substance P and met-enkephalin-arg-phe, respectively. In the rat and Callithrix jacchus, opioid immunoreactivity is present in nerve fibres but not in glomus cells. Immunoreactivity to other peptides, such as neurotensin, cholecystokinin, neuropeptide Y, and galanin, is found only in one or two particular species. Neurotensin immunolabelling occurs in beagle dog glomus cells, which are known to lack substance P. Cholecystokinin immunoreactivity is present in glomus cells of dog and Callithrix, and co-exists with chromogranin A, neuropeptide Y, and substance P. Substance P appears to exist in both carotid body glomus cells and nerve fibres. Substance P immunoreactivity is present in glomus cells of all species investigated, except dog. Coexistence of substance P and calcitonin gene-related peptide (CGRP) is demonstrated in nerve fibres of the guinea pig carotid body, which originate in the petrosal and jugular ganglia. Other peptides visualized immunohistochemically in mammalian carotid body nerve fibres are vasoactive intestinal peptide and neuropeptide Y. The functional significance of the various peptides present in the carotid body is discussed.     

Survivin蛋白和AIF在胃癌组织中的表达

目的探讨Survivin和AIF在胃癌组织中的表达及对胃癌诊断和预后评估的价值。方法应用免疫组化方法检测我院56例胃癌手术标本的Survivin和AIF的表达情况,并对这些患者进行随访。结果Survivin和AIF在胃癌细胞中表达阳性率分别为71.43%(40/56)和66.07%(37/56),Survivin在癌旁组织和正常胃组织中无表达,AIF在癌旁组织和正常胃组织中低表达;Survivin和AIF在胃癌中表达无显著相关性;Survivin及AIF的阳性和阴性组1、3、5年累积生存率均有统计学差异。结论Survivin和AIF在胃癌组织中的阳性表达率明显高于癌旁组织和正常胃组织,可以作为判断胃癌患者预后的指标,但二者的表达无明显相关性。     

BAK和BAG-1蛋白在结直肠癌中的表达及其意义

目的探讨BAK和BAG-1蛋白在结直肠癌中的表达及其意义。方法采用免疫组化SP法对80例结直肠癌、40例结直肠腺瘤及25例癌旁正常结直肠黏膜组织中BAK和BAG-1蛋白的表达进行检测,并分析其与结直肠癌临床病理特征之间的相关性。结果 BAK和BAG-1蛋白在结直肠癌中过度表达,表达率分别为72.5%(58/80)和80%(64/80),且BAG-1蛋白与肿瘤的分化程度呈负相关,BAK蛋白与肿瘤的分化程度呈正相关。有淋巴结转移的结直肠癌,其BAG-1蛋白的表达率为96.7%(29/30),无淋巴结转移的结直肠癌,其BAG-1蛋白的表达率为70%(35/50),两者相比差异有统计学意义(P<0.01)。结论 BAK和BAG-1蛋白的过度表达与肿瘤的分化程度密切相关,且BAG-1蛋白的过度表达还与肿瘤的淋巴结转移密切相关;结直肠癌的发生机制可能涉及BAK和BAG-1调节环路中多个基因的异常。     

乙型肝炎病毒X蛋白表达与肝细胞脂肪变性的关系及其机制

目的探讨乙型肝炎病毒X蛋白(Hepatitis Bvirus X protein,HBx)在肝组织中的表达与慢性乙型肝炎(Chronic hepatitis B,CHB)合并肝脂肪变性的关系及其可能的机制。方法对经病理证实的18例CHB及18例CHB合并肝脂肪变性的病例标本进行血脂、肝功能、血清病毒学及病理组织学指标分析,免疫组化法检测肝组织HBx、肝X受体(Liver X receptorα,LXRα)和脂肪酸合成酶(Fatty acid synthase,FAS)的表达。结果两组病例生化指标、HBV血清标志物及HBV-DNA载量、肝组织炎症活动度分级及纤维化程度分期的构成比差异均无统计学意义(P>0.05);HBx、LXRα和FAS表达的阳性率差异也无统计学意义(P>0.05);但CHB合并肝脂肪变性组HBx、LXRα和FAS的表达强度明显高于CHB组,且差异有统计学意义(P<0.05);在CHB合并肝脂肪变性组中,HBx与LXRα、LXRα与FAS及HBx与FAS的表达呈正相关。结论 HBx在CHB肝组织中广泛表达,在CHB合并肝脂肪变性组的表达明显增强,提示HBx可能通过诱导LXRα进而刺激FAS表达上调,参与CHB合并肝细胞脂肪变性的发生。     

宫颈腺样囊性癌临床病理特征及免疫组织化学表型

【摘要】目的 探讨4例宫颈腺样囊性癌（Adenoid cystic carcinoma, ACC）临床病理及免疫组化特征，并总结回顾文献。 方法 收集北京协和医院外科病理数据库及会诊数据库中诊断为宫颈腺样囊性癌病例共计4例，总结其临床病理特征、治疗方法及预后等；并采用免疫组化方法对石蜡组织标本进行检测；总结回顾中国人群宫颈腺样囊性癌特征。 结果 4例宫颈腺样囊性癌平均年龄为，患者多为绝经后妇女，就诊主要症状为阴道流血，肿瘤类型多为外生性肿物；病理学上，3例患者表现为宫颈腺样囊性癌合并侵袭性鳞癌，1例为宫颈腺样囊性癌单一组分；腺样囊性癌的主要生长方式为筛状；免疫组织化学表达方式支持其导管腺上皮及肌上皮两种组成成分：CK7在导管上皮中表达阳性，P63、SMA在肌上皮中表达；腺样囊性癌特征性标记物MYB在所有病例中均强阳性表达。结论 宫颈腺样囊性癌是一种罕见的特殊类型的腺癌，常合并其他类型的宫颈肿瘤，免疫组化表型与其他部位的腺样囊性癌相同，但预后相对较差，术后可辅以放疗及化疗，因其易早期复发及转移，建议对患者进行密切的随诊。     

Predictive value of p63, ki‐67, and survivin expression in oral leukoplakia: A tissue microarray study

The aim of this study was to analyze the immunohistochemical expression of survivin, ki‐67, and p63 in oral leukoplakic lesions, histopathologically differentiated into dysplastic and nondysplastic. A tissue microarray containing 57 samples of biopsies from clinically classified lesions, such as leukoplakia, was immunolabeled for survivin, ki‐67, and p63. Samples were scored for percentage of positively stained. Scores were designated as follows: low = less than 25% of positive cells; and high = more than 25% of positive cells. On performing histopathological diagnosis, 20 dysplastic lesions and 37 nondysplastic lesions were seen, in which female patients (56.1%) were predominant with an average age of 58.27 years. The study showed a high expression of 37.5% for survivin, 43.7% for ki‐67, and 88.2% for p63 in dysplastic lesions. However, there was a high expression of 16.7% for survivin, 16.7% for ki‐67, and 92% for p63 in nondysplastic lesions. There is a positive correlation of expression among the three antibodies. In the association of immunoreactivity, in both dysplastic and nondysplastic lesions, increased expression of survivin reflects on the increased expression of ki‐67, and there is an overexpression of p63. In leukoplakia, the expression of survivin associated with that of ki‐67 reinforces the assumption that all these lesions are potentially malignant, regardless of histopathology; and the overexpression of p63 may indicate carcinogenic potential. These findings may help in the treatment of patients with this type of lesion.     

Segmentation of digitized histological sections for quantification of the muscularized vasculature in the mouse hind limb

Immunohistochemical tissue staining enhances microvasculature characteristics, including the smooth muscle in the medial layer of the vessel walls that is responsible for regulation of blood flow. The vasculature can be imaged in a comprehensive fashion using whole‐slide scanning. However, since each such image potentially contains hundreds of small vessels, manual vessel delineation and quantification is not practically feasible. In this work, we present a fully automatic segmentation and vasculature quantification algorithm for whole‐slide images. We evaluated its performance on tissue samples drawn from the hind limbs of wild‐type mice, stained for smooth muscle using 3,3'‐Diaminobenzidine (DAB) immunostain. The algorithm was designed to be robust to vessel fragmentation due to staining irregularity, and artefactual staining of nonvessel objects. Colour deconvolution was used to isolate the DAB stain for detection of vessel wall fragments. Complete vessels were reconstructed from the fragments by joining endpoints of topological skeletons. Automatic measures of vessel density, perimeter, wall area and local wall thickness were taken. The segmentation algorithm was validated against manual measures, resulting in a Dice similarity coefficient of 89%. The relationships observed between these measures were as expected from a biological standpoint, providing further reinforcement of the accuracy of this system. This system provides a fully automated and accurate means of measuring the arteriolar and venular morphology of vascular smooth muscle.