神经烯醇化酶、突触素和神经纤维丝蛋白在人胚胎心脏组织中的表达

目的探讨神经烯醇化酶(NSE)、突触素(SYN)和神经纤维丝蛋白(NF)在人胚胎心脏组织不同发育阶段的分布特征。方法应用免疫组织化学法,检测第2~4个月胎龄段共16份人胚胎心脏组织内NF、NSE和SYN蛋白的表达,分析其变化规律。结果第2~4个月龄段,NF、NSE和SYN蛋白在人胚胎心脏组织内均有阳性表达。随着胎龄的增大,NF、NSE和SYN在心脏组织内阳性表达强度值逐渐降低。第2个月龄时,NSE、NF和SYN蛋白呈少量阳性表达,阳性表达强度值分别是86.79±7.75、133.03±13.61和114.32±11.12。第3个月龄时,NSE、NF和SYN阳性表达强度值分别是81.89±9.62,119.91±11.70和93.13±13.63。第4个月胎龄时,NSE、NF和SYN阳性表达强度值分别是72.18±11.97,107.02±10.89和91.17±13.81。应用One-Way ANOVA和LSD-t统计学分析第2~4个月龄段人胚胎心脏组织内NSE、NF和SYN蛋白的各自阳性表达强度值,P0.05。结论第2~4个月龄段,NSE、NF和SYN均在人胚胎心肌组织内表达和呈现特定的分布规律,随胎龄增大,心肌组织内NF、NSE和SYN的表达强度逐渐增强。     

神经纤维丝蛋白、神经烯醇化酶和突触素在人胚胎舌组织中的表达

目的 探讨神经纤维丝蛋白（NF）、神经烯醇化酶(NSE)和突触素(SYN)在人胚胎舌组织不同发育阶段的分布特征。 方法 应用免疫组织化学法,检测第2～4个月胎龄段共16份人胚胎舌组织内NF、NSE和SYN蛋白的表达,分析其变化规律。 结果 第2～4个月龄段,NF、NSE和SYN蛋白在人胚舌组织内均有阳性表达。随着胎龄的增大,NF、NSE和SYN在舌组织内阳性表达数量增多,表达强度逐渐增强。第2个月龄时,NF、NSE和SYN蛋白呈少量弱阳性表达,阳性表达强度值分别是135.83±24.62、136.57±15.23和139.84±21.40。第3个月龄时,NF、NSE和SYN阳性表达强度值分别是96.04±23.37、94.89±22.52和90.65±21.08。第4个月胎龄时,NF、NSE和SYN阳性表达强度值分别是79.02±20.90、76.78±21.27和83.43±25.90。应用 One-Way ANOVA和 LSD-t统计学方法,分析第2～4个月龄段人胚胎舌组织内NF、NSE和SYN蛋白的各自阳性表达强度值,P＜0.01。 结论 第2～4个月龄段,人胚胎舌组织内NF、NSE和SYN的表达强度值随胎龄增大而降低,它们均参与调控人胚胎舌内神经系统和舌肌的分化发育。     

Gastroenteropancreatic neuroendocrine tumors. A histochemical and immunohistochemical study of epithelial (keratin proteins, carcinoembryonic antigen) and neuroendocrine (neuron-specific enolase, bombesin and chromogranin) markers in foregut, midgut, and hindgut tumors

Thirty-four gastroenteropancreatic (GEP) neuroendocrine tumors were evaluated for expression of epithelial (keratin, carcinoembryonic antigen [CEA] and neuroendocrine (neuron-specific enolase, chromogranin, bombesin) markers, and results were correlated with histologic patterns and histochemical staining. Tumors of mixed pattern (insular or trabecular with glandular areas) predominated. CEA localization corresponded to staining for mucin, with polarized apical or lumenal staining in glandular areas. Four trabecular midgut carcinoids, however, revealed diffuse cytoplasmic staining for CEA. Staining for keratin proteins was present in 68% of tumors. Bombesin immunoreactivity was demonstrated in 60% of GEP neuroendocrine tumors, indicating that bombesin positive metastatic tumors may not be predominantly of pulmonary origin, as previously suggested. Chromogranin was a sensitive marker for identifying normal gastrointestinal neuroendocrine cells that were not demonstrated by staining for neuron-specific enolase. Chromogranin was present in most neuroendocrine tumors, but was absent from three of five rectal carcinoids in keeping with the distinctive profile of hormonal and silver staining in these tumors. All GEP neuroendocrine neoplasms expressed both neuroendocrine and epithelial markers, supporting their derivation from endodermal epithelium.     

Expression of gamma-subunit of enolase, neuron-specific enolase, in human non-neuroendocrine tumors and derived cell lines

The gamma-subunit of 2-phospho-D-glycerate hydrolyase, E.C. 4.2.1.11 (enolase), neuron-specific enolase (NSE), is present at high concentrations in neurons and neuroendocrine cells and has therefore recently been introduced as a marker for neuroendocrine tumors. By the indirect methods, immunocytochemistry and radioimmunoassay, NSE has been detected also in some nonneuroendocrine tumors, a finding that could reflect technical artifacts or the capacity for NSE expression in nonneuroendocrine tumor cells. This paper reports on the expression of NSE in human neuroendocrine and nonneuroendocrine tumor specimens and in a panel of permanent human cell lines, by using a direct (enzymatic) and an indirect (radioimmunoassay) method for determination of NSE. We detected NSE in all tested tumor specimens and neuroendocrine tumor cell lines and in a majority (21 of 24) of the nonneuroendocrine tumor cell lines. In general, neuroendocrine tumor specimens and derived tumor cell lines contained more NSE than the nonneuroendocrine tumor specimens and cell lines. However, some of the cultured hematopoietic cell lines (T leukemia and Epstein-Barr virus immortalized B lymphoblastoid cell lines) had NSE levels comparable to those found in some neuroblastoma and small-cell lung carcinoma cell lines. We conclude that NSE is not exclusively expressed in neuroendocrine tumor cells.     

An ultrastructural and immunohistochemical evaluation of cytodifferentiation in neuroblastic tumors

Twenty-six cases of neurogenic tumors consisting of 2 ganglioneuromas (GN), 8 ganglioneuroblastomas (GNB), and 16 neuroblastomas (NB) were studied to evaluate their cytodifferentiation. Ultrastructurally, a moderate to large number of neuritic processes and high density neurosecretory granules (NSG) were found in all cases of GN and well-differentiated GNB, in two-thirds of poorly differentiated GNB, in about a half of rosette-fibrillary NB, and in no case of round-cell NB. All GN and GNB had tumor cells which were positive for both chromogranin and neurofilaments. Of the 16 cases of NB, 12 were positive for chromogranin, and 13 and 15 were positive for Mr 200,000 and 68,000 neurofilament polypeptides, respectively. However, both markers appeared mainly in tumor cells maturing toward neuroblasts. Electron microscopy was helpful for the diagnosis of undifferentiated NB in those cases immunohistochemically negative for chromogranin or neurofilaments. We conclude that ultrastructural and immunohistochemical examinations are useful for the morphologic assessment of the degree of maturation of neuroblastic tumors.     

Expression of neurofilament triplet proteins in human neural tumors. An immunohistochemical study of paraganglioma, ganglioneuroma, ganglioneuroblastoma, and neuroblastoma.

Intermediate filaments which are specific to neural cells, ie, neurofilaments, consist of three subcomponents--68, 150, and 200 kd. Thirty human neural tumors were examined for the presence of these three subcomponents by means of their monospecific antisera. All 8 paragangliomas contained cells that were positive for the 68-kd component, but only 5 of them had cells positive for the 150-kd and 200-kd components. All 4 ganglioneuromas and 11 ganglioneuroblastomas contained cells that reacted with antibodies to all three components. All 7 neuroblastomas had cells reacting with antibody to 68 kd, but only 3 of them had cells that reacted with antibodies to 150 kd and 200 kd. In each case, the number of positive cells depended on the antibody used. The largest number reacting with antibody to 68 kd and the smallest with antibody to 200 kd. Furthermore, it was possible to detect tumor cells in which the 68-kd subcomponent existed by itself, but no tumor cells in which the 150-kd or 200-kd subcomponent existed alone could be detected. These results seem to indicate that antibody to the 68-kd component is sufficiently discriminating to be applied diagnostically.     

An immunohistochemical study on the distribution of glial fibrillary acidic protein, S-100 protein, neuron-specific enolase, and neurofilament in medulloblastomas

In order to clarify the differentiation of medulloblastomas, the authors studied on the morphological features and immunohistochemical expression of glial fibrillary acidic protein (GFAP), S-100 protein, neuron-specific enolase (NSE), and neurofilament (NF) in 31 medulloblastomas. GFAP was detected only in a small number of tumor cells of 5 medulloblastomas; S-100 protein in both small tumor cells and some so-called spongioblastic cells in 16 medulloblastomas; NSE in the more abundant tumor cells and the matrix in 28 medulloblastomas; NF in a few tumor cells of 12 medulloblastomas; GFAP and NF in 2 medulloblastomas, but each of them in different tumor cells. These results suggest that medulloblastomas have a capacity of differentiation along neuronal and/or glial lines. The conventional morphological markers of differentiation in medulloblastomas such as spongioblastic cells and Homer Wright rosettes were not necessarily compatible with expression of immunohistochemical markers such as GFAP or NF. NSE and S-100 protein seem less valuable markers of differentiation because they were detected in both neuronal and glial elements. But NSE, which was observed in most medulloblastomas, might have a value as a marker for medulloblastomas.     

An immunohistochemical demonstration of neuron-specific enolase in the Merkel cells of the frog taste organ

The Merkel cells in the taste organ of the frog were investigated by immunohistochemistry using neuron-specific enolase (NSE) antiserum. NSE-immunoreactivity was found exclusively in the Merkel cells lying at the base of the taste organ. The distribution and the profiles of the NSE-immunoreactive Merkel cells coincided with serotonin-containing cells previously reported at the same place.     

Pituitary adenomas. An immunohistochemical study of hormone production and chromogranin localization.

Tumors from 42 surgically resected pituitaries and from 13 autopsy cases were studied immunohistochemically with polyclonal antisera to 7 anterior pituitary hormones and with a newly developed monoclonal antibody directed against human chromogranin for evaluation of the distribution of chromogranin in normal and neoplastic pituitaries. In addition, a prospective study was done for assessment of the prevalence, morphology, and endocrine cell types of pituitary tumors in 100 autopsy subjects. When these 55 pituitary adenomas were examined with monoclonal antibody (LK2H10) directed against human chromogranin, selective staining of normal adenohypophyseal cell types and pituitary tumors was observed. Most null-cell adenomas (12/14) were positive for chromogranin, whereas all prolactin (PRL)-producing adenomas (19/19) were negative. Growth hormone (GH) adenomas were focally positive (9/9). All oncocytomas (2/2), 1 thyrotropin (TSH) adenoma, and a follicle-stimulating hormone/luteinizing hormone adenoma were positive for chromogranin. One or more adenomas were present in 14% of the autopsy cases. The tumors occurred most frequently in patients in the fifth through the seventh decades of life. Immunohistochemical staining of 13 adenomas revealed 1 TSH, 1 ACTH, and 4 PRL-producing tumors, whereas 7 other tumors, which were null-cell or undifferentiated adenomas, failed to stain for any of the seven principle pituitary hormones. These results indicate that antibody LK2H10 to human chromogranin is useful in the immunohistochemical characterization of pituitary adenomas. Incidental pituitary microadenomas from autopsy-derived pituitaries most commonly produce PRL, or they belong to the null-cell or undifferentiated tumor group.     

Expression of vitamin B12 R-binder in breast tumors. An immunohistochemical study

Vitamin B12 R-binder, a vitamin B12-specific binding protein, was demonstrated by immunohistochemistry in 103 cases of breast tumors of various types and grades. In normal breast tissues, R-binder was found on the luminal surface of interlobular, intralobular, and small terminal ducts. R-binder localization was observed in all fibroadenomas and phyllode tumors, in 90% of the benign epithelial tumors, and in 33% of the carcinomas. Among malignant tumors, 43% of the ductal carcinomas expressed the R-binder, whereas none of the lobular or mucinous carcinomas were positive. In invasive ductal carcinomas, R-binder expression was demonstrated more often in well-differentiated lesions.     

Widespread p53 overexpression in human malignant tumors. An immunohistochemical study using methacarn-fixed, embedded tissue.

p53 is a nuclear protein believed to play an important role, through mutation and overexpression, in the progression of human malignant tumors. The authors employed a monoclonal antibody, 1801, and investigated overexpression of p53 in a series of 255 malignant and benign tumors, using deparaffinized sections of methacarn-fixed tissue. Overall, immunohistochemically detected p53 overexpression was found in 39% of malignant tumors, with considerable variation within individual tumor types (34% of breast carcinomas, 92% of ovarian carcinomas, 33% of soft tissue sarcomas). Homogenous, heterogenous, and focal immunostaining patterns were noted. With rare exceptions, no immunostaining of any benign tumors was noted. No immunostaining was found in adjacent, benign tissues, or in a series of fetal tissues. This is the first demonstration of widespread p53 overexpression in alcohol-fixed, embedded tissue and confirms the major role played by p53 in human malignancies.     

Expression of novel neuroendocrine marker insulinoma-associated protein 1 (INSM1) in genitourinary high-grade neuroendocrine carcinomas: An immunohistochemical study with specificity analysis and comparison to chromogranin,synaptophysin, and CD56

Confirmation of genitourinary high-grade neuroendocrine carcinomas (GU-HGNECs) often requires immunohistochemical staining. Here we evaluated a novel neuroendocrine marker, insulinoma-associated protein 1 (INSM1), in GU-HGNECs with comparison to chromogranin, synaptophysin and CD56. Immunohistochemical expression of INSM1, chromogranin, synaptophysin, and CD56 was evaluated in 39 GU-HGNECs using full tissue sections [4 in kidney, 28 in urinary bladder, and 7 in prostate; 31 small cell carcinomas (SmCCs), 6 large cell neuroendocrine carcinomas (LCNECs), 2 mixed SmCC-LCNECs]. In 33 SmCCs/components, INSM1 showed similar sensitivity (93.9 %) to chromogranin (87.8 %), synaptophysin (93.9 %) and CD56 (87.8 %), and stained a similar percentage of tumor cells (52 %) to chromogranin (49 %) and CD56 (52 %), but lower than synaptophysin (87 %) (p < 0.0001). In 8 LCNECs/components, INSM1 is similar to chromogranin, synaptophysin or CD56 in sensitivity (62.5 %, 62.5 %, 75 %, 62.5 %, respectively) and the mean percentage of positively stained tumor cells (21 %, 44 %, 48 %, 37 %, respectively). INSM1 is more sensitive for SmCCs than LCNECs (93.9 % vs. 62.5 %, p = 0.015). INSM1 showed 97.4 % specificity upon analyzing 273 genitourinary non-neuroendocrine tumors on tissue microarrays. Our study indicates that INSM1 is a sensitive marker for genitourinary HGNECs with high specificity. For genitourinary SmCCs, INSM1 shows similar sensitivity to chromogranin, synaptophysin and CD56 but stains a lower percentage of tumor cells than synaptophysin. For genitourinary LCNECs, INSM1 showed similar sensitivity to chromogranin, synaptophysin and CD56. INSM1 is more sensitive for genitourinary SmCCs than LCNECs. Our result and literature review indicate that whether INSM1 is more sensitive than conventional neuroendocrine markers for HGNECs depends on the tumor primary sites.     

Prostatic origin of tumors. An immunohistochemical study

An immunoperoxidase technic was used to localize prostatic acid phosphatase in a variety of primary and metastatic neoplasms. The aim was to explore the histogenesis of tumors affecting the prostate gland and to demonstrate the prostatic origin of metastases in various sites. A highly specific antiserum to prostatic acid phosphatase was raised in rabbits, and the peroxidase-antiperoxidase procedure was carried out on formalin-fixed paraffin-embedded routine pathology material. All specimens from the 37 cases of known primary and metastatic prostatic carcinomas stained positively for prostatic acid phosphatase, regardless of their histologic differentiation. None of the specimens from the 44 cases of proven nonprostatic primary and metastatic tumors stained positively for prostatic specific acid phosphatase. The data suggest that demonstration of prostatic acid phosphatase by the immunoperoxidase technic is a practical, sensitive, and specific test for the prostatic origin of an otherwise unclassifiable primary or metastatic neoplasm.     

Demonstration of differential immunohistochemical localization of the neuron-specific enolase antigen in rat pinealocytes

A refined method for the immunohistological demonstration of neuron-specific enolase (NSE) on 1- to 2-μm Epon-812 section gave characteristic staining of cerebral and cerebellar neurons. This method has made it possible to obtain a more detailed characterization of the heterogeneity of rat pinealocytes in the superficial portion of the rat pineal complex. Thirty adult male rats have been studied, five of which were used in a photometric analysis of the distribution of NSE. Pinealocytes stained either intensely or weakly for the NSE antigen and exhibited an uneven distribution within a given region. Further analysis of the gland revealed a distal to proximal decrease in stain intensity. It is suggested that the more strongly stained cells, being concentrated distally, are under sympathetic control.     

Increased chromogranin A and neuron-specific enolase in rats with chronic nonbacterial prostatitis induced by 17-beta estradiol combined with castration

Although chronic nonbacterial prostatitis (CNBP) is a common diagnosis in middle-aged men, the etiology of this disease remains poorly understood. Neuroendocrine cells play an important role in the neuroendocrine regulation of the prostate, and chromogranin A (CgA) and neuron-specific enolase (NSE) are regarded as classic markers of neuroendocrine cells. This study aimed to determine CgA and NSE levels in a CNBP rat model to evaluate the role of neuroendocrine cells in the pathogenesis of CNBP. For developing a CNBP rat model, we examined the ability of 17-beta estradiol and surgical castration alone or in combination to induce CNBP. Histologic inflammation of the prostate was assessed in CNBP-induced rats by hematoxylin-eosin staining, whereas CgA and NSE protein levels were assessed by immunohistochemistry, Western blot analysis, and enzyme-linked immunosorbent assays. Our results showed that 17-beta estradiol combined with castration successfully induced CNBP and that CgA and NSE levels were increased in the prostate of CNBP rats as compared to those without CNBP. These findings indicate that the neuroendocrine regulation mediated by neuroendocrine cells may be involved in the pathogenesis of CNBP.     

Alpha-fetoprotein in germ cell tumors. An immunohistochemical study

Total 40 cases of testicular and ovarian germ cell tumors and one case of extragonadal germ cell tumor were studied for the presence of alphafetoprotein (AFP) by indirect immunoperoxidase technique. All seminomas (7 cases) and teratomas (13 cases) were negative for AFP; while 85% of the pure embryonal carcinomas, (E.C.) all pure yolk sac tumors (Y.S.T.) (7 cases) and all embryonal carcinoma and yolk sac components in mixed tumors were AFP positive. Immunostaining of tumor marker appeared to help only in differentiating seminomatous and nonseminomatous tumors and hence does not provide any additional information for classification of these tumors.     

Extrauterine mixed mesodermal tumors. An immunohistochemical study.

Mixed mesodermal tumors are uncommon outside the uterus. Nine extrauterine mixed mesodermal tumors (eight ovarian and one extragenital) were selected for histochemical and immunoperoxidase study. In eight cases, both epithelial and mesenchymal elements were malignant (chondroid in six, rhabdomyoid in four, and osteoid in two). One ovarian tumor was an adenosarcoma. All cases were stained with periodic acid-Schiff with and without diastase and for alpha 1-antitrypsin, myoglobin, keratin, vimentin, muscle-specific actin, and alpha 1-antichymotrypsin, by using the avidin-biotin-immunoperoxidase method. The periodic acid-Schiff-positive, diastase-resistant droplets in several of the tumors showed peripheral alpha 1-antitrypsin positivity. Keratin delineated epithelial areas well in seven cases, and rhabdomyoid differentiation was confirmed with myoglobin in four cases. However, squamous elements in one tumor were falsely positive for myoglobin. We concluded that despite occasional cross-reactivity, carefully interpreted immunoperoxidase stains can be useful in distinguishing epithelial and mesenchymal elements in these tumors.     

Expression of P53 protein in normal,dysplastic, and malignant gastric mucosa: An immunohistochemical study

Mutations in the p53 nuclear oncogene are the most frequent genetic abnormalities encountered in human malignancies. Using the polyclonal antibody CM-1, we have examined the expression of the p53 oncoprotein immunohistochemically in archival material of normal, dysplastic, and malignant gastric mucosa. Abnormal expression of this protein was not observed in biopsies of normal gastric tissue (n=30) but was detected in 22 of the 36 gastric cancers analysed (61 per cent). Nuclear staining was diffuse in 15 of the positive cancer cases, the remaining seven showing a more varied heterogeneous staining pattern. Abnormal p53 protein was not detected in mild (n=14) or moderate (n=16) gastric dysplasia but was present in 3 out of 15 severe dysplasia cases. The results suggest that expression of the p53 oncoprotein is a common finding in gastric cancer and occurs as a late event in the malignant transformation process.