正常胃肠道上皮粘液性质及其显示方法

应用组织化学方法包括PAS,AB/PAS,HID/AB,OR/AB,PAPS,mPAS,PB/KOH/PAS,PAT/KOH/PAS,PATB/KOH/PAS观察正常胃肠道粘膜分泌粘液的成分.对各种组织化学方法进行了比较,并对国内尚无报告的新方法进行了介绍.在正常胃粘膜表面上皮及胃窦腺体以分泌中性粘液为主.小肠主要分泌氮乙酰化唾液酸粘液,大肠上皮分泌以硫酸粘液及氧乙酰化唾液酸粘液为主.     

Mucin histochemistry of cystitis glandularis and primary adenocarcinoma of the urinary bladder

We describe the histochemical properties of mucin in three cases of cystitis glandularis affecting the urinary bladder. Of particular importance is the positive staining reaction that was obtained with the periodate borohydride/potassium hydroxide/PAS (PB/KOH/PAS) technique, indicating the presence of O-acetylated sialic acids. This reaction has been regarded as unique to the epithelial mucins in the normal terminal ileum and large intestine. Therefore, cystitis glandularis represents a true mucous metaplasia of large intestinal type. A case of primary adenovillous carcinoma of the bladder associated with cystitis glandularis also produced O-acetylated sialomucin, whereas a primary adenocarcinoma of urachal origin did not. The demonstration of O-acetylated sialic acid producing glandular epithelium in the bladder cannot be assumed to represent a metastasis from a colorectal cancer. The PB/KOH/PAS staining technique may provide a means of distinguishing between primary adenovillous and primary urachal carcinoma of the bladder.     

Mucins in breast carcinoma

Mucins produced by breast carcinoma have been shown to be neutral and sialomucins. Colonic mucins were found to be sulfated with an o-acylated sialomucin component that was reported by Culling to be specific to normal colonic epithelium and colonic adenocarcinoma. This type of molecule gives a positive staining after potassium hydroxide/periodate borohydride (KOH/PBT) treatment. We studied the types of mucin in five medullary carcinomas, 10 infiltrating duct carcinomas, and 10 infiltrating lobular carcinomas of the breast. Twenty-two of 25 cases were positive for neutral mucins; 14 of 25 were positive for sialomucin. Sulfated mucins were found in five of 10 cases of infiltrating lobular carcinoma, and in six of 10 cases there was a positive periodic acid-Schiff reaction following KOH/PBT. This study illustrates the presence of sulfated mucin and o-acylated sialomucin in infiltrating lobular carcinoma of the breast. Thus, mucin stains cannot be used in differentiating metastatic breast carcinoma from colonic carcinoma. The significance of this finding and its relation to tumor histogenesis, typing, and biologic behaviors require further analysis.     

Mucinous (colloid) adenocarcinomas secrete distinct O-acylated forms of sialomucins: a histochemical study of gastric, colorectal and breast adenocarcinomas

AIMS: Mucinous (colloid) adenocarcinomas represent a distinct group of tumours defined by the presence of large amounts of extracellular mucins. By using histochemical methods, we analysed mucins secreted by mucinous versus non-mucinous adenocarcinomas and looked for differential secretion profiles. METHODS AND RESULTS: Sixty-four adenocarcinomas were studied (23 colorectal, 17 gastric, and 24 breast tumours). Thirty-two tumours were of the colloid type. The following methods were applied to paraffin tissue sections: (i) Alcian blue (pH 2.5) and periodic acid-Schiff (PAS); (ii) high iron diamine and Alcian blue (pH 2.5); (iii) periodic acid borohydride, potassium hydroxide, and PAS; (iv) periodic acid-thionine Schiff, potassium hydroxide, and PAS; and (v) periodic acid-borohydride and PAS. Most adenocarcinomas secreted acidic mucins, with sialomucins predominating over sulfomucins, except for non-mucinous adenocarcinomas of the breast which showed predominant neutral mucins. All mucinous adenocarcinomas contained C9-O-acyl sialic acid as mono, di(C8,C9)-, or tri(C7,C8,C9)-O-acyl forms. Acidic mucins secreted by the majority of non-colloid adenocarcinomas consisted of non-O-acylated sialomucins. CONCLUSIONS: C9-O-acylation of sialic acid is a characteristic feature of mucinous adenocarcinomas and can be readily detected by histochemical methods.     

HISTOCHEMICAL DEMONSTRATION OF COLONIC TYPE MUCIN IN GLANDULAR METAPLASIA and ADENOCARCINOMA OF THE HUMAN URINARY BLADDER

Mucin in normal, metaplastic and neoplastic tissue specimens of the human urinary bladder was examined histochemically by the periodic acid-borohydride/potassium hydroxide/periodic acid-Schiff (PB/KOH/PAS) reaction and paradoxical concanavalin A (Con A) staining, both of which have been reported to detect mucins that are specific for the lower gastrointestinal tract. Normal mucosa, Brunn's nest, squamous metaplasia, transitional cell carcinoma, and squamous cell carcinoma produced only small amount of mucin and were negative for both PB/KOH/PAS and labile class III by Con A staining. However, several sections of mucin-producing metaplastic bladder lesions and many samples of adenocarcinoma arising from vesical mucosa and urachus contained PB/KOH/PAS-positive and/or labile class III mucin. Thus the colonic type mucin appeared in some materials of both benign and malignant mucin-producing lesions of the urinary bladder. The findings would be related to the embryogenesis of this organ.     

Histochemical demonstration of colonic type mucin in glandular metaplasia and adenocarcinoma of the human urinary bladder

Mucin in normal, metaplastic and neoplastic tissue specimens of the human urinary bladder was examined histochemically by the periodic acid-borohydride/potassium hydroxide/periodic acid-Schiff (PB/KOH/PAS) reaction and paradoxical concanavalin A (Con A) staining, both of which have been reported to detect mucins that are specific for the lower gastrointestinal tract. Normal mucosa, Brunn's nest, squamous metaplasia, transitional cell carcinoma, and squamous cell carcinoma produced only small amount of mucin and were negative for both PB/KOH/PAS and labile class III by Con A staining. However, several sections of mucin-producing metaplastic bladder lesions and many samples of adenocarcinoma arising from vesical mucosa and urachus contained PB/KOH/PAS-positive and/or labile class III mucin. Thus the colonic type mucin appeared in some materials of both benign and malignant mucin-producing lesions of the urinary bladder. The findings would be related to the embryogenesis of this organ.     

O-acetylated sialic acid variants in intestinal glandular metaplasia of the urinary tract

Two cases of chronic cystitis, two of chronic ureteritis, one of chronic pyelitis and two transitional cell carcinomas of the urinary tract showed glandular metaplasia or focal mucin production. Positive staining for O-acetylated sialic acid variants (sites 7 and 8) detected by the PB-KOH-PAS method was found in the two cases of cystitis, in one of chronic ureteritis and in one of chronic pyelitis. Positive staining was observed only in areas showing an intestinal type of metaplasia and in which goblet cells and argyrophil cells were also identified. These findings taken together with earlier studies on ovarian cystadenomas lead the authors to believe that O-acetylated variants are markers of an intestinal type of epithelium. Evaluation of O-acetylated sialomucins may be useful in further studies on the histogenesis of primary adenocarcinomas of the bladder.     

Mucinous adenocarcinoma of the colon - a histochemical study

Colorectal carcinoma is a major cause of cancer associated with a high rate of morbidity and mortality in the western world. One of the pathologic features considered to be important for prognostic is mucin production. Many authors confirmed that colon carcinomas with high mucin content tend to re-occur locally and carry a poor prognosis. For histochemical evaluation of mucin content, we investigated 149 patients who underwent surgical resection of sporadic colon adenocarcinomas, all over a 5-year period. For histological classification we used the WHO recommendation (2000) and to be more accurate we sub-classified mucinous adenocarcinomas by morphometrical analysis in three categories: pure mucinous, with extracellular mucin more than 80% of the tumoral volume; mixed type, with 50-80% extracellular mucin; and mixed type with less than 50% extracellular mucin. For histochemical investigation, we used stains such as: mucicarmine, PAS ÷ Alcian Blue and High Iron Diamine ÷ Alcian Blue. Our study proved the predominance of mixed mucinous adenocarcinomas with less than 50% extracellular mucin, followed by the pure mucinous type. From the biochemical composition's point of view, the predominant cases were those with acidic mucins, especially in pure mucinous adenocarcinomas (>90%), while those with mixtures of acidic and neutral mucins were present in 62% of the cases. In addition, our study showed the prevalence of sialomucins over sulphomucins (68%), particularly in pure mucinous adenocarcinomas (77%). Clinical pure mucinous forms were detected mainly in advanced stages, but in terms of lymph node metastasis rate, they were secondary after mixed type with 50-80% extracellular mucin.     

Histochemistry and electron microscopy in the diagnosis of small mucous cells in carcinoma of the stomach

One hundred and two cases of poorly differentiated adenocarcinoma were obtained from 71 surgical and 31 biopsy specimens of the stomach. These tumours produced both intracellular and extracellular mucins, which were demonstrated to be neutral mucins, N-acetyl sialomucins, sulphomucins and O-acetyl sialomucins. The majority of tumours (85.3%) secreted two or more kinds of mucins simultaneously, but only a small proportion (14.7%) produced a single kind of mucin. Ultrastructurally, three types of mucin granules were identified; these differed in shape and density. Tumour cells containing intracellular mucins or mucin granules were designated as mucous cancer cells and further subdivided into small, large and goblet mucous cells; tumour cells having neither mucins nor glandular differentiation were designated as undifferentiated cancer cells. These histochemical distinctions, supported by the electron microscopical observations, may be useful to demonstrate early malignant foci in gastric biopsies.     

Retrospective histochemical study of mucosubstances in adenocarcinomas of the gastrointestinal tract

Use of the PATS/KOH/PAS and PB/KOH/PAS techniques and the diamine method (Spicer 1965) together with the demonstration of metachromasia with toluidine blue pH 3.0, for the study of carbohydrates in a retrospective study of gastric and colorectal adenocarcinomas, disclosed an increase in the sulphate content and the absence of O-acylated sialomucins in the majority of gastric adenocarcinomas. In the colorectal tumours there is no change in the degree of sulphation normally present in this area, but the degree of acylation of the sialic acid in adenocarcinoma is very different to that of normal mucosa and appears related to the degree of differentiation of the tumour. The mucosa adjacent to tumours from both areas were also studied, as well as gastric intestinal metaplasia.     

Mucin histochemistry of mucous carcinomas of breast and colon and non-neoplastic breast epithelium

Although epithelial mucin has been demonstrated in ordinary mammary duct carcinomas, it is formed profusely in the uncommon mucous carcinomas. A histochemical study of mucous carcinomas of the breast and colon showed that both types produce similar mucins, viz, acid and neutral, although the mammary carcinomas produce more neutral mucin and small amounts of sulphated mucins are produced by colonic tumours. Similar studies on sections of normal breast and cystic mammary dysplasia have shown that intracellular neutral mucin was present in the ducts and lobules, and that intracellular acid mucin was present within cells of the terminal ducts and lobules. It was demonstrated enzymatically that the acid mucins produced by the normal and neoplastic tissues is sialic acid. Hence it can be shown that the mucin produced by mucous carcinomas is of epithelial rather than connective tissue type. It is contended that mucous carcinoma of breast arises by metaplasia of duct cells that have already undergone malignant change and not from the lobules of the breast.     

Mucin profiles in ulcerative colitis with dysplasia and carcinoma

Mucin secretion was assessed in Crohn's colitis, in ulcerative colitis with regeneration, dysplasia and carcinoma and in non-colitic adenocarcinoma. The high iron diaminealcian blue (HID–AB) and periodate borohydride–saponification periodic acid Schiff (PB–KOH–PAS) techniques were used to demonstrate sulphomucins and sialomucins, and O-acylated sialomucins respectively. There was mucosal hyperplasia and increased sialomucin secretion in Crohn's disease, quiescent and active ulcerative colitis. In colitis with carcinoma inflamed mucosa away from the tumour had increased sialomucins as had colitis with dysplasia. They did not differ statistically from each other or from colitic controls without cancer. Dysplastic crypts frequently secreted sulphomucins and the increased sialomucins were in transitional-like glands in the surface fronds or adjacent to the dysplasia. A comparative study of the HID–AB technique gave total correct qualitative allocation of individual quantitatively assessed crypts. Routine HID–AB staining did not aid the recognition of dysplasia in ulcerative colitis. With the PB–KOH–PAS technique colorectal adenocarcinoma showed a significant diminution in O-acylated sialomucins compared with its adjacent mucosa. Mucosal dysplasia in ulcerative colitis displayed a similar trend in O-acylated sialic acid variants, differing with respect to age- and sex-matched colitic controls. The PB–KOH–PAS technique may be of help in assessing mucin secretion in ulcerative colitis as a guide to the evolution of malignancy.     

Villous adenoma of the bladder.

A case of villous adenoma of the bladder associated with cystitis glandularis of intestinal type is described. Only three villous adenomas have been reported to date, of which two were also accompanied by cystitis glandularis. The lesion can be confused with polypoid hyperplasia in cystitis glandularis. Neutral mucins, acidic sulphomucins, and sialomucins were identified within the villous adenoma and adjacent areas of cystitis glandularis. It is suggested that the villous adenoma may form an intermediary stage in the development of some of the primary adenocarcinomas of the bladder arising in metaplastic intestinal mucosa.     

Concanavalin A binding sites in fetal, adult, transitional, and malignant rectosigmoid mucosa

We studied mucin histochemistry in 25 rectosigmoid adenocarcinomas and in the transitional mucosa adjacent to these tumors using standard techniques for the detection of neutral and acid sialomucins and sulfomucins and the paradoxical concanavalin A (Con A) stain. This histochemical procedure selectively detects residues of mannose in glycoproteins exposed to brief steps of oxidation and reduction. Those techniques were also used to study histologically normal mucosa of specimens with carcinoma, normal rectosigmoid mucosa of patients without inflammatory or neoplastic bowel disease, hyperplastic rectal polyps, and rectosigmoid mucosa of human fetuses. Normal mucosa and hyperplastic polyps mainly contained sulfomucins and did not display Con A binding activity with any of the variants of the stain. In contrast, fetal, transitional, and malignant mucosa predominantly showed sialomucins and although not reactive with the standard Con A sequence, displayed binding activity for the lectin after short oxidative-reductive steps. These results provide further evidence that transitional and malignant mucosa produce markedly abnormal mucins whose histochemical patterns represent a re-emergence of the fetal type found during development. The principles of the paradoxic Con A reaction may be applied to unmask lectin binding activity in apparently unreactive sites.     

Mucin histochemistry in primary adenocarcinoma of the urinary bladder (of urachal or vesicular origin) and metastatic adenocarcinoma originating in the colorectum

In order to evaluate the mucin histochemistry of primary adenocarcinomas (PA) of the urinary bladder and metastatic adenocarcinoma (MA) originating in the colorectum, 52 PA and nine MA were examined. It was determined that the percentage of cases in which more than 25% of the tumor was stained by each of the following: (i) Alcian blue pH 2.5 periodic acid-Schiff (AB-PAS); (ii) high iron diamine-AB (HID-AB); (iii) periodic acid-sodium borohydride-potassium hydroxide-PAS (PA-SB-PH-PAS); (iv) galactose oxidase- Schiff (GOS); and (v) paradoxical concanavalin A stain (PCS). For PA, the values obtained were: 75% of cases (blue, AB-PAS), 85% (magenta, AB-PAS), 71% (black, HID-AB), 75% (blue, HID-AB), 0% (PA-SB-PH-PAS), 19% (GOS), 8% (class II concanavalin A (Con A)-reactive mucin)), and 0% (class III Con A-reactive mucin). For MA, the corresponding values were 33, 22, 0, 11, 0, 0, 11, and 0%, respectively. A higher percentage of PA than MA cases showed staining in AB-PAS for acidic and neutral mucins, in HID-AB for sialo- and sulfomucins, and in GOS for terminal beta-galactose and beta-N-acetylgalactosamine. PA and MA were significantly different in terms of both frequency of staining with AB-PAS and frequency of staining with HID-AB. However, the overlap was such that in practice, it might be difficult, if not impossible, to use mucin histochemistry to inform a differential diagnosis. In view of the differences in AB-PAS and HID-AB positivity between PA and MA, we speculate that MA (originating in the colorectum) may have undergone structural distortion affecting the production and/or secretion of neutral mucins and acidic mucins (sialo- and sulfomucins) during metastasis or invasion.     

Mucin profiles in the mucosa adjacent to large bowel non-adenocarcinoma neoplasias

Morphological features and mucin secretion patterns were investigated in the colonic mucosa adjacent to or overlying mesenchymal or primary epithelial neoplasias, other than adenomas and adenocarcinomas. The material included 15 cases of non-adenocarcinoma tumours examined during 1978-1981. Increased sialomucins and morphological features similar to those described in the so-called 'transitional' mucosa adjacent to primary colorectal adenocarcinomas were observed in only two cases. In contrast our previous studies have demonstrated 'transitional' profiles in 98% of adenocarcinomas.     

Expression of MUC5AC apomucin in transitional cell carcinomas of the urinary bladder and its possible role in the development of mucus-secreting adenocarcinomas

The histogenesis of primary nonurachal mucus-producing adenocarcinomas of the urinary bladder including signet ring cell carcinomas remains to be elucidated, since the normal bladder contains neither columnar nor mucus-secreting glandular epithelium. Based upon the assumption that adenocarcinomas may develop secondarily from pre-existent transitional cell carcinomas (TCC) by a metaplastic process, it was the purpose of the current immunohistochemical study to analyze whether urothelial carcinomas are capable of secreting MUC5AC apomucin, using the monoclonal antibody 45MI. This antibody has been initially demonstrated to strongly react with the mucus-producing columnar cells of the surface gastric epithelium, recognizing a specific epitope located on the peptide core of glycoproteins as major components of mucins. Nine of 64 uniformly differentiated papillary (14.1%) and 5 of 66 nonpapillary (solid) TCC with a uniform urothelial differentiation (7.6%) expressed the MUC5AC antigen, yielding an overall incidence of 10.8%. Transitional cell carcinomas with a focally altered cellular and structural differentiation (squamous cell, pseudoglandular, true glandular and mixed differentiation) stained positively in a substantially higher percentage of 43.8% (21 of 48 cases). A positive immunoreactivity was also observed in 3 of 19 mixed transitional cell and nonurothelial carcinomas. The tumor-associated resurgence of normally cryptic MUC5AC antigenic determinants in transitional cell carcinomas is considered as a re-expression of oncofetal antigenicity, probably as a result of the embryologic origin of the urinary bladder from the pluripotent tissues of the cloacal endoderm and the mesodermal wolffian ducts. Our findings may help to better understand the histogenetic development of mucus-secreting vesical adenocarcinomas from pre-existent urothelial carcinomas.     

A histochemical comparison of the O-acylated sialic acids of the epithelial mucins in ulcerative colitis, Crohn''s disease, and normal controls.

Two histochemical techniques, the PAT/KOH/PAS and the PBT/KOH/PAS, were used to investigate the side chain O-acyl substitution patterns of the sialic acids of the colonic epithelial mucins in cases of ulcerative colitis and Crohn's disease. In both diseases there was, as compared to normal, a reduction in the proportion of sialic acids O-acylated at C7C8, the reduction being greater in ulcerative colitis. Further, there appeared to be an association between the severity of the disease and the reduction in the staining of O-acylated sialic acids. This relationship was more marked in ulcerative colitis. In some cases of both diseases there was evidence for epithelial mucins containing predominantly C7-substituted sialic acids. This study has confirmed our previous conclusion that, in Crohn's disease of the terminal ileum, the disease is associated with an increase in the proportion of sialic acids bearing side chain substituents.     

Mucous metaplasia of epithelial cells in fibrocystic disease of the breast. A histochemical study

Mucous metaplasia of breast epithelium was studied by histochemical procedures in cases of fibrocystic disease. Epithelial mucins consist almost entirely of non-sulfated sialomucins. A slight amount of sulfated sialomucins was identified occasionally while neutral mucins seems to be absent. Abundant glycogen granules are contained in the cytoplasm of metaplastic cells. The secretion of sialomucins may be associated with a loss of cell antigenicity. In this case, the survival of metaplastic epithelium supports the hypothesis of the possible immunological nature of the disease.     

Secondary neoplasms of the bladder are histological mimics of nontransitional cell primary tumours: clinicopathological and histological features of 282 cases

AIMS: The incidence, anatomical localization and histological appearances of secondary neoplasms of the urinary bladder are described, with emphasis on the points of distinction from primary tumours. METHODS AND RESULTS: A retrospective study of cases at the Royal Hospitals Trust yielded a total of 282 secondary bladder neoplasms, representing 2.3% of all malignant bladder tumours in surgical specimens. The commonest primary sites were the colon (21% of secondary neoplasms), prostate (19%), rectum (12%) and cervix (11%). Most tumours from these sites reached the bladder by direct spread. The most common sites of origin of tumours metastatic to the bladder were stomach (4.3% of all secondary bladder neoplasms), skin (3.9%), lung (2.8%), and breast (2.5%). Secondary tumour deposits were almost always solitary (96.7%), and 54% were located in the bladder neck or trigone. Histologically, 54% of secondary tumours were adenocarcinomas. Immunohistochemical staining patterns with prostate-specific acid phosphatase, prostate-specific antigen, carcinoembryonic antigen, chromogranin and neurone-specific enolase were similar in primary vesical and urachal adenocarcinomas and secondary adenocarcinomas from the gastrointestinal tract. CONCLUSIONS: The incidence of secondary bladder tumours is comparable to that of nontransitional cell primary tumours. Few secondary tumours have distinctive histological features, hence knowledge of the history and clinical investigations are particularly important in their diagnosis.