KiSS-1 overexpression as an independent prognostic marker in hepatocellular carcinoma: an immunohistochemical study

The KiSS-1 gene has been reported to play an important role as a metastasis suppressor gene in various human malignancies. However, there is little information about its possible role in hepatocellular carcinoma (HCC). In this study, we evaluated the prognostic significance of the expression of KiSS-1 and its receptor AXOR12 in 142 HCC tissue specimens by immunohistochemistry. By using a cutoff level of 50%, immunoreactivity of KiSS-1 and AXOR12 was found in 6 (4%) and 11 (8%) HCCs. The expression of KiSS-1 and AXOR12 in HCC correlated with each other (r = 0.42, p < 0.0001) and with the expression in corresponding, surrounding liver tissue (both r = 0.35, p < 0.0001). Positive AXOR12 immunoreactivity in HCC correlated with advanced pT-stage of tumors and low tumor grading (r = 0.18, p = 0.032; r = −0.18, p = 0.029). High KiSS-1 expression in HCC had a statistically significant influence on diminished disease-free and overall survival in uni- (p = 0.006 and p = 0.002) and multivariate analysis (r = 2.874, p = 0.027 and r = 2.913, p = 0.026). In this study, we report for the first time that elevated KiSS-1 expression level in HCC correlates with worsened clinical outcome, as an independent prognostic marker for the aggressiveness of HCC.     

Retinoblastoma and p53 gene expression related to relapse and survival in human breast cancer: an immunohistochemical study.

Inactivation of tumour suppressor genes may be an important aetiological factor in many human cancers including breast. In a study of 197 breast cancer patients, tumour tissue was snap-frozen at the time of surgery and immunohistochemical labelling for p53 protein and retinoblastoma (Rb) gene product carried out using an indirect immunohistochemical technique. Tumours were scored by two independent observers for the intensity of nuclear staining for each antibody. Expression of p53 protein showed a significant association with a shorter time to relapse (P = 0.03) and death (P = 0.02) (log rank test). p53 expression did not correlate with nodal status but showed a significant association with high tumour grade (P = 0.001). Rb gene expression showed no relationship to relapse or survival but loss of expression showed a significant correlation with positive lymph node status. The manner by which these proteins might act to determine tumour behaviour remains to be established.     

Melanosis of the prostate: an immunohistochemical study

We report here a case of prostatic melanosis examined immunohistochemically. The patient was a 70-year-old man with prostatic hyperplasia. Six foci with brown pigmentation were found, not only in stromal cells but also in glandular epithelial cells among many pieces of prostatic tissue. Immunohistochemically, only pigment-laden stromal cells were obviously positive with antibodies to S-100 protein and melanoma (HMB45), but not for CD57, while epithelial cells were only positive for CD57 after depigmentation with bleaching. This is the first report demonstrating immunopositivity of stromal cells for antimelanoma antibody (HMB45), which can label reactive or dysplastic but not normal melanocytes. These results clearly demonstrated that pigment-laden stromal cells are neuroectodermal in origin, and probably reactive or dysplastic but not normal melanocytes.     

WT1 as a complementary marker of malignant melanoma: an immunohistochemical study of whole sections

AIMS: To test the usefulness of WT1 as a diagnostic aid in melanoma diagnosis and prognostication. METHODS AND RESULTS: Benign naevi, Spitz naevi, dysplastic naevi and melanoma in situ, primary epithelioid, spindle cell and desmoplastic melanoma, and metastatic melanoma biopsy specimens were collected. Primary melanoma cases were grouped using the 2003 Tumour Node Metastasis classification. Cases were examined using haematoxylin and eosin and WT1-stained sections. The presence and pattern of WT1 expression were recorded. Benign naevi were uniformly negative with WT1. The majority of Spitz naevi expressed WT1 (83%). The dysplastic naevi/in situ group showed expression of WT1 in 35% of cases. The majority of primary epithelioid melanoma cases showed WT1 expression (88%). The prevalence of expression by each T category was similar, with T1 = 90%, T2 = 75%, T3 = 95% and T4 = 90%. All spindle cell and desmoplastic melanoma cases showed WT1 expression (100%). Ninety per cent of metastatic melanoma cases expressed WT1. CONCLUSIONS: WT1 is a useful ancillary diagnostic tool in routine melanoma diagnosis. WT1 expression in primary melanoma is unrelated to tumour depth. Its usefulness is limited by the fact that most Spitz naevi express WT1, up to one-third of dysplastic naevi can express WT1 and not all melanomas demonstrate its expression.     

Cellular diversity of human placental stem villi: an ultrastructural and immunohistochemical study

The aim of the study was to investigate the distribution and differentiation of cell types in the stroma of human placental stem villi (SV). A total of 14 human term placental tissues were studied. Double immunolabeling was performed for desmin-vimentin, desmin--smooth actin and vimentin--smooth actin. Cytokeratin 7, proliferating cell nuclear antigen immunolabeling was also performed. Parallel tissue samples were examined by transmission electron microscopy. HSCORE was performed for the semi-quantitative analysis of distribution of cells in the stroma of SV. Vimentin-labeled cells were mostly distributed in the subtrophoblastic area. Desmin-vimentin double immunolabeling was mainly localized in the triangular area and to a lesser degree in the perivascular area and vessel walls (p=<0.001). However, desmin- smooth actin labeling was observed predominantly in the vessel wall and perivascular area. Vimentin- smooth actin immunoreactivity was significantly stronger in the triangular and perivascular areas compared to the vessel walls (p=0.003).

Ultrastructurally, cells in the stroma of SV were mesenchyme cells, reticulum cells, fibroblasts, myofibroblasts, smooth muscle cells, and Hofbauer cells, filamented and vacuolated cells. The differentiation of myofibroblasts in the triangular and perivascular areas may play a role in maturation of SV and villous contractility, modulation of the intervillous space and this may have effects on maternofetal placental circulation.     

Expression of p53, pRB,and p16 in lung tumours: a validation study on tissue microarrays

Tissue microarrays have been created from 326 lung tumours, including 173 squamous cell carcinomas (SCCs) and 132 adenocarcinomas (ADs). In order to evaluate the usefulness of this microarray series, the expression of p53, p16, and Rb proteins was compared by immunohistochemistry on both the tissue microarrays and the corresponding whole sections for all 326 tumours. The presence of replicate punches improved both the yield and the concordance of data relative to the whole section results, so that the consensus score from the replicates agreed with the whole section result in more than 90% of informative tumours. The large number of tumours in this series also allowed significant differences in protein expression patterns to be detected between SCC and AD, the major subtypes of non-small cell lung carcinoma (NSCLC). SCC had higher levels of p53 staining (67% vs 52% in AD) and substantially increased p16 loss (SCC 75%, AD 53%) combined with greater retention of pRB expression (SCC 86% vs 67% in AD). The strong inverse correlation between p16 and pRB seen in SCC was essentially absent in AD. This study represents the largest single immunohistochemical survey of protein expression for p53, p16, and RB in NSCLCs.     

Three cases of sclerosing lipogranuloma: an immunohistochemical study

Sclerosing lipogranuloma (SLG) is a rare disease that occurs in the male genital region. We report here three cases of SLG, including two of Y-shaped lesions in the penile base forming an intrascrotal mass, as well as a patient with a mass in the epidydimal region. All three lesions histologically exhibited formation of multiple granulomas consisting of multinucleated giant cells and epithelioid cells, in the fibroadipose tissue or around the epidydimis, in association with eosinophil infiltration. Membranocystic changes were found within the cytoplasm of multinucleated cells. In the two patients with scrotal lesions, membranocystic changes were positive for diastase-PAS reaction and on Sudan black B staining in dewaxed sections. CD68 staining was strongly positive in multinucleated giant cells and epithelioid cells. Most of the lymphocytes infiltrating the lesions were T cells associated with some S-100-positive dendritic cells. T-cell mediated immune reaction appears to be important in the histogenesis of SLG. The histogenesis is generally considered to be a foreign-body reaction to degenerated or damaged fatty tissue or lipids, but no apparent causative factors can be identified in some reported cases. All three patients in the present study had a good clinical course after biopsy or surgical excision. We thus report three cases of SLG including an unusual case in the epidydimal region, with some considerations regarding the histogenesis of SLG.     

Expression of the HNK-1 carbohydrate epitope in human retina and retinoblastoma

Summary Fifty formalin-fixed and paraffin-embedded retinoblastoma specimens and five normal human eyes were studied with the monoclonal anti-Leu-7 antibody, directed against the HNK-1 carbohydrate epitope that is shared by human natural killer cells and many neuronal, glial and neuroectodermal cells. The laboratory method was a sensitive immunohistochemical staining procedure, and neuroectodermal tumours that usually express this epitope were used as positive controls. In the human retina, Müiller cell membranes were positively stained, but additional staining of neuronal cells was not excluded at the light microscopical level. A positive cytoplasmic reaction was also seen in ciliary and retinal pigment epithelial cells. All but one intraocular retinoblastomata studied contained cells staining positively for the HNK-1 epitope, but these cells were probably not neoplastic. Although positive reaction has previously been reported in three retinoblastomata, the present results suggest that positive cells are derived from entrapped and infiltrated retina. Staining of adjacent sections against leukocyte common antigen suggested that the positively staining cells were not natural killer cells.This study was supported by a grant from the Emil Aaltonen Foundation     

胚胎发育不良性神经上皮瘤与皮质发育不良

目的观察胚胎发育不良性神经上皮瘤（DNT）的病理形态学及免疫组织化学特点,探讨其与皮质发育不良之间的关系以及组织来源。方法应用光镜和免疫组织化学EnVision法对14例DNT进行观察分析,并对患者进行长期随访。结果肿瘤位于颞叶的有11例,镜下由神经元和神经胶质成分混合构成,9例可见“特异的胶质神经元结构”。1例为简单型,8例为复杂型,5例为非特异型。11例标本充足的病例10例伴有皮质发育不良改变,表现为分子层和（或）白质内异位神经元数量增多（7例）,分子层内可见成行的外颗粒细胞层残留（4例）,脑皮质的结构异常（10例）,以及出现异常形态的神经细胞。免疫组织化学染色少突胶质样细胞（OLC）均呈现Olig2的免疫反应性,部分OLC表达nestin、微管相关蛋白2、神经丝蛋白、胶质纤维酸性蛋白,但神经元核抗原呈阴性。癫痫控制结果Ⅰ级12例,Ⅱ级2例,无肿瘤复发。结论DNT与皮质发育不良关系密切,应用免疫组织化学染色有助于DNT和皮质发育不良的诊断。     

Expression and immunohistochemical detection of Nesfatin-1 in the gastrointestinal tract of Casertana pig

In this study, we report nesfatin-1 immunoreactivity in the gastrointestinal tract of Casertana breed pig. The newly discovered anorexigenic peptide nesfatin-1 has been shown to possess physiological relevance in regulating food intake and energy homeostasis at a central level, although evidence has been accumulating that it may also play important functions at a more local gastroenteric level. Nesfatin-1 immunoreactive endocrine cells have been detected in the gastric fundus and ileocecal valve. Nesfatin-1 immunopositive neurons and nerve fibers have been observed mainly in the enteric plexuses. Western blot analysis confirmed the immunohistochemical observations, showing immunoreactive bands in all analyzed gastrointestinal tracts with the exception of the rectum. Nesfatin-1 immunodetection in the swine digestive system reinforces the importance of the role played by nesfatin-1 at the gastrointestinal level and sustains the necessity to study the role of this peptide in the regulation of food intake in farm species for which weight gain is essential for optimizing production.     

Malignant rhabdoid tumour of the uterus: an immunohistochemical and ultrastructural study

Summary Malignant rhabdoid tumours (MRTs) are highly aggressive neoplasms which most frequently occur in the kidney of young children. Several cases of primary MRT occurring in extra-renal sites have been reported, particularly in the soft tissues. We report a case of primary MRT of the uterus, a very rare site for this neoplasm, with morphological, immunohistochemical and ultrastructural features corresponding to restrictive morphological criteria for MRT. The possible differential diagnoses were considered.     

Neuropeptides in the human dorsal vagal complex: An immunohistochemical study

The distribution of twelve biologically active neuropeptides, i.e., thyrotropin-releasing hormone, corticotropin-releasing factor, pro-opiomelanocortin-derived peptides (adrenocorticotropic hormone, β-endorphin, α-melanocyte-stimulating hormone), leucine-enkephalin, dynorphin A, dynorphin B, cholecystokinin, substance P, galanin and calcitonin gene-related peptide, was examined by immunohistochemistry in the human dorsal vagal complex including the nucleus of the solitary tract, the dorsal motor nucleus of the vagus and the area postrema. Immunoreactivity of all the twelve neuropeptides was found widely distributed in the various subdivisions of the nucleus of the solitary tract, showing a unique distribution for every peptide. Neuronal cell bodies immunostained with leucine-enkephalin, galanin and dynorphin B were found in this region. There were no immunopositive perikarya for any of the peptides in the other structures studied. Fibers containing galanin, corticotropin-releasing factor, substance P, dynorphin B, thyrotropin-releasing hormone and calcitonin gene-related peptide were observed at a relatively high density in the nucleus of the solitary tract. In the same structure, a moderately dense network of fibers immunostained with dynorphin A, cholecystokinin and leucine-enkephalin, but only solitary pro-opiomelanocortin-derived peptides-containing fiber fragments were observed. In the dorsal motor nucleus of the vagus the most prominent network of fibers was found to contain thyrotropin-releasing hormone, galanin and substance P. In contrast to these, no β-endorphin immunoreactivity was detected. The area postrema contained only moderate to low densities of galanin-, substance P-, calcitonin gene-related peptide-, dynorphin B- and cholecystokinin-immunoreactive fibers.     

Expression of glypican 3 in hepatoblastoma: an immunohistochemical study of 65 cases

Glypican 3 is a heparin sulfate proteoglycan bound to the cell surface that is theorized to participate in cell signaling, resulting in embryonic cell growth and differentiation. The GPC3 gene is mutated in Simpson-Golabi-Behmel syndrome, whose features include numerous developmental abnormalities, tissue overgrowth, and an increased risk for embryonal malignancies, including hepatoblastoma. GPC3 has been detected in hepatic stem cells and was recently identified as one of the most overexpressed genes in hepatoblastoma by microarray analysis. The purpose of this study was to analyze the expression of GPC3 in a large series of hepatoblastoma using immunohistochemistry to assess its use as a diagnostic marker. The GPC3 immunoreactivity was semiquantitatively evaluated in 65 cases of hepatoblastoma. In addition, histologic patterns in each tumor were individually assessed for immunoreactivity. All 65 hepatoblastomas had cytoplasmic immunoreactivity for GPC3 with greater than 90% of cases showing strong, diffuse positivity. There was no reactivity in benign liver tissue. Fetal, embryonal, and small cell undifferentiated patterns were diffusely positive in almost all cases, whereas mesenchymal and teratoid patterns were nearly all negative. The high GPC3 expression consistently demonstrated in this study suggests that GPC3 may play a role in the tumorigenesis of hepatoblastoma.     

Primary retroperitoneal mucinous cystoadenocarcinomas: an immunohistochemical and molecular study

Special immunohistochemical stains for the identification of gastroenteropancreatic antigens in two cases of primary retroperitoneal mucinous cystoadenocarcinomas (PRMC) show that these tumours have patterns similar to ovarian mucinous tumours. Markers of pyloric type gastric mucosa differentiation (M1, cathepsin E, concavavalin A, pepsinogen II) are mostly positive in benign and borderline areas with endocervical type differentiation, while immunoreactivity for intestinal cell markers (M3SI and CAR-5) and for DU-PAN-2 is present mainly in frankly malignant areas, regardless of differentiation type. DNA analysis shows a point mutation of K-ras oncogene at codon 12 (GGT to CGT) in one case. The immunohistochemical and genotypic similarity of PRMC and ovarian mucinous tumours may indicate similar mechanisms in their histogenesis.     

Mutational screening of the RB1 gene in Italian patients with retinoblastoma reveals 11 novel mutations

Retinoblastoma (RB, OMIM#180200) is the most common intraocular tumour in infancy and early childhood. Constituent mutations in the RB1 gene predispose individuals to RB development. We performed a mutational screening of the RB1 gene in Italian patients affected by RB referred to the Medical Genetics of the University of Siena. In 35 unrelated patients, we identified germline RB1 mutations in 6 out of 9 familial cases (66%) and in 7 out of 26 with no family history of RB (27%). Using the single-strand conformational polymorphism (SSCP) technique, 11 novel mutations were detected, including 3 nonsense, 5 frameshift and 4 splice-site mutations. Only two of these mutations (1 splice site and 1 missense) were previously reported. The mutation spectrum reflects the published literature, encompassing predominately nonsense or frameshift and splicing mutations. RB1 germline mutation was detected in 37% of our cases. Gross rearrangements outside the investigated region, altered DNA methylation, or mutations in non-coding regions, may be the cause of disease in the remainder of the patients. Some cases, e.g. a case of incomplete penetrance, or variable expressivity ranging from retinoma to multiple tumours, are discussed in detail. In addition, a case of pre-conception genetic counselling resolved by rescue of banked cordonal blood of the affected deceased child is described.     

嗅神经母细胞瘤的光镜、电镜和免疫组织化学观察

报道了１１例嗅神经母细胞瘤的光镜、免疫组化及其中２例超微结构观察结果。１１例嗅神经母细胞瘤中１０例可见瘤细胞排列成大小不等的巢状或条索状；８例有Ｈｏｍｅｒ－Ｗｒｉｇｈｔ菊形团，５例见Ｆｌｅｎｅｒ菊形团（其中４例同时伴有Ｈｏｍｅｒ－Ｗｒｉｇｈｔ菊形团）；８例可见瘤细胞之间由细胞突起形成的嗜酸性纤维样背景。电镜下１例瘤细胞内见神经原纤维，２例均有神经内分泌颗粒，并对其颗粒进行了形态测量。多种抗体免疫组化染色：ＮＳＥ１１例均为阳性，Ｖｉｍｅｎｔｉｎ，Ｓ－１００、Ｋｅｒａｔｉｎ各有４例、３例和１例阳性，ＮＦ，ＥＭＡ，ＣＥＡ，ＬＣＡ均为阴性。文中对嗅神经母细胞瘤的病理形态及免疫组化特征在诊断中的意义进行了讨论。     

Early development of chick embryo respiratory nervous system: an immunohistochemical study

The extrinsic and intrinsic respiratory nervous systems receive specific contributions from the vagal and sympathetic components. Using specific markers for vagal and sympathetic structures, we studied the distribution patterns of immunoreactivity to galanin (GAL), pituitary adenylate cyclase-activating polypeptide-27 (PACAP) and the tachykinin substance P in extrinsic and intrinsic nerve of chick embryo respiratory system, during development from the very early age to hatching. All peptides studied appeared in the intrinsic and extrinsic nervous systems early. We found substance P in both the vagal and sympathetic systems, PACAP in vagal components alone and GAL mainly in the sympathetic system. The intrinsic nervous system showed high immunoreactivity for all peptides studied. These data accord with the well known early trophic functions that peptides have on the development of nervous networks and modulatory activity on the intrinsic nervous system. The GAL again proves to be the main peptide in chick embryo sympathetic respiratory system.     

Orexin 1 receptor in the seminiferous tubules of boar testis: An immunohistochemical study

Orexin receptor 1 (OX₁R) and orexin receptor 2 (OX₂R) are two G-protein-coupled receptors that bind their ligands, orexin A (OXA) and B (OXB), with different affinities. The male genital system represents an important target for OXA, which appears to play a role in the control of steroidogenesis and germ cell development in the testis. It is known that among domestic breeding animals, in the boar the number of Leydig cells is very high and OXA appears to have stimulatory activity on testosterone production. In this study, we aimed to evaluate the presence of OX₁R in the boar testis in order to extend our knowledge concerning the distribution and a potential functional role of the orexinergic system in the male reproductive tract of farm animals. The presence of OX₁R immunopositive cells in seminiferous tubules of the boar testis enables us to hypothesize a possible role of OXA on male germ cells cycle in pig. Further investigations, involving functional and ultrastructural analysis, may contribute to our understanding of the role of orexins in the boar genital system.     

伴微乳头状分化的胃肠道腺癌的临床病理学特征和免疫组织化学研究

目的 探讨伴微乳头状分化的胃肠道腺癌(GAMPD)的临床病理特征及上皮细胞膜抗原(EMA)、胰岛素样生长因子Ⅱ mRNA结合蛋白-3(IMP3)和E-cadherin表达的临床意义.方法 对73例GAMPD进行临床病理学分析,并应用免疫组织化学EnVision法检测EMA、IMP3和E-cadherin 在GAMPD中的表达.结果 在73例GAMPD中,微乳头状癌分化成分所占比例为5%～70%,多见于中分化腺癌.与普通腺癌比较,GAMPD在转移阳性淋巴结和TNM分期上存在明显差异.淋巴结转移性微乳头状癌成分与原发灶微乳头状癌成分含量呈正相关.微乳头状癌细胞簇间质面EMA表达阳性率为52.1%(38/73).EMA+组好发于胃(P=0.018),EMA+ 组阳性淋巴结平均数量为(6.6±5.8)枚,多于EMA-组(3.8±4.7)枚,两组差异具有统计学意义(P=0.029),其余临床病理学参数EMA+组与EMA-组比较差异无统计学意义.IMP3在EMA+组GAMPD中的表达阳性率为86.8%(33/38),显著高于普通腺癌(P＜0.05).而E-cadherin在GAMPD中的阳性表达率则显著低于普通腺癌(P＜0.00).结论 GAMPD是一种在组织学形态、免疫表型和生物学行为上不同于胃肠道普通腺癌的特殊类型,侵袭性强、预后差.免疫组织化学联合检测EMA、E-cadherin和IMP3的表达对GAMPD的病理诊断和预测预后有帮助.

Abstract：

Objective To study the clinicopathologic features and immunophenotype of gastrointestinal adenocarcinomas with a micropapillary pattern differentiation(GAMPD). Methods Seventy-three eases of GAMPD arising in gastrointestinal tract were retrospectively reviewed.Immunohistochemical study for epithelial membrane antigen(EMA), insulin-like growth factorⅡmRNAbinding protein-3(IMP3)and E-cadherin was performed.Results Amongst the 73 cases studied,the micropapillary pattern accounted for 5% to 70% of the tumor component.It was often seen in a background of moderately differentiated adenocarcinoma.As compared with conventional adenocarcinoma,nodal metastasis was more frequently observed and the TNM tumor stage was statistically higher in GAMPD.The occurrence of micropapillary component in metastatic lymph nodes positively correlated with the proportion of mieropapillary pattern in primary lesions.EMA staining on the stroma-facing surface of tumor micropapillae was demonstrated in 52.1%(38/73)of the cases.As compared with EMA-negative GAMPD, EMA-positive GAMPD was more in the stomach(P=0.018).and with more metastatic lymph nodes(6.6 4-5.8 vs 3.8 ±4.7, P=0.029).The rate of IMP3 expression in EMA-positive GAMPD was 86.8%(33/38),which was higher than that in conventional adenocarcinoma.In contrast.the rate of E-cadherin expression in GAMPD was lower than that in conventional adenocarcinoma.Conclusions GAMPD is a distinctive variant of gastrointestinal adenocarcinomas and different from conventional adenocarcinoma in tumor morphology.immunophenotype and biologic behavior.It carries an aggressive clinical course and poor prognostic outcome.Immunohistochemical study for EMA.IMP3 and E-cadherin would be helpful in the diagnosis and prognostic evaluation of GAMPD.     

Epithelial cell adhesion molecule expression in pituitary adenomas: an immunohistochemical study

An important amount of data correlating the expression of epithelial cell adhesion molecule (Ep-CAM) with cellular proliferation and de-differentiation could directly contribute to carcinogenesis. The aim of this study is to evaluate prognosis relevance of Ep-CAM expression in a group of pituitary adenomas. Epithelial cell adhesion molecule, proliferating cell nuclear antigen, and microvascular density labeling indices in pituitary adenomas were determined by immunohistochemistry on tissue samples obtained from each adenoma after surgery. We evaluated 45 adenomas. Sixty-two percent were nonsecretor adenomas and 37.8% were secretor tumors. Immunohistochemistry was scored for immunoexpression of Ep-CAM (cytoplasmic, membrane, and mixed pattern). Proliferating cell nuclear antigen and vascular density (CD34) labeling indices were assessed. Statistical significance was observed between Ep-CAM cytoplasmic immunoreactions (P = .000) and higher proliferating cell nuclear antigen (P = .001) in secretor adenomas compared with nonsecretor tumors. Vascular density labeling indices did not show statistical significance. Therefore, Ep-CAM could be evaluated to distinguish secretor and nonsecretor pituitary adenomas. These suggest that the markers could predict the growth potential of individual pituitary adenomas.