大肠癌及癌前病变的粘液组化研究

应用粘液组化方法AB—PAS、HID—AB、OR—AB、PAT—KOH—PAS、PATB—KOH—PAS显示大肠上皮分泌粘液的类型。结果发现:大肠慢性炎、大肠克隆病及炎性息肉、增生性息肉、幼年性息肉分泌粘液类型与正常粘膜相似(P>0.05),而伴有异型增生的溃疡性结肠炎、管状腺瘤、绒毛状腺瘤、家族性息肉、腺瘤癌变、大肠癌及癌旁粘膜粘液分泌性质与量均有变化。在这些病变中氮乙酰化及含有邻位羟基的氧乙酰化粘液阳性率明显高于正常粘膜(P<0.05)。其检出阳性率与异型增生的程度有关。笔者认为应用粘液组化方法分析大肠癌前病变粘液性质对于判断粘膜上皮恶性倾向提供了有价值的手段。     

347例大肠息肉及其癌变的粘液组化和免疫组化观察

对大肠息肉及其癌变347例的粘液组化和223例的免疫组化研究显示,它们的粘液分泌改变和四种结肠癌相关抗原表达与其组织学类型、体积大小和不典型增生程度相关,并且PAT/KOH/PAS粘液组化染色显示氮乙酰化、侧链C_7、C_9位氧乙酰化唾液酸粘液分泌可能代表了息肉的恶变倾向,结肠癌单克隆抗体CL-2、CL-4的免疫组化染色对判断腺瘤癌变有较大参考价值。     

对一种新粘液组化染色法的评价

近年来,粘液组织化学染色在诊断胃肠道肿瘤及癌前病变方面已显示出重要价值。由于方法本身不够完善,临床应用还很局限。1985年,Reid等报道了一种新的粘液组化方法即:KOH/AB 1.0/PAPS方法,该法能同     

用AB/PAs、HID、CEA法提高胃粘液细胞癌淋巴结转移检出率的研究

用AB(pH2.5)/PAS、HID及CEA(ABC法)对48例胃粘液细胞癌及其淋巴结进行染色观察,对照HE染色,结果表明应用上述方法染色可提高淋巴结转移检出率33.1%,其中AB(pH2.5)/PAS法显示了很强的特异性和敏感性,较HID和CEA法优越。淋巴结微小转移灶多为PAS阳性细胞,说明幼稚的癌细胞分泌中性粘液、侵袭力强、容易转移并先发生转移。CEA标记结果显示本肿瘤有分化越差、越幼稚其CEA含量越少的现象。     

三重染色在结肠癌病理诊断中的应用

本文介绍三重染色技术在同一切片上应用,其中用结肠癌单克隆抗体MC_3,显示结肠癌细胞内的肿瘤相应抗原;用AB或AB/PAS法显示结肠癌组织内的酸性、中性及混合性粘蛋白对32例原发性结肠癌的观察结果,三染阳性率达96.87％,证实MC_3-McAb原发性结肠癌的特异性强。MC_3染色在各型结肠癌阳性细胞浆内及腺腔缘处可见到棕黄色颗粒及弥漫性阳性物质,同时可见到大量PAS染色呈红色的中性粘液物质,这些细胞多位于细胞底2/3位置,正常少见。AB染色显示蓝色酸性粘液,在癌组织间质及癌旁间质混合性粘液呈紫色片状、糊状及正规则型,正常结肠组织中未见到此现象。 本文建立三重染色法,对结肠癌的研究提供免疫组化及组化相结合的方法。     

胃粘膜肠化生及粘液癌分类的探讨

对50例良性及50例恶性病变幻肠化生及固有病变进行了光镜观察,探讨了肠化生与粘液癌的分类及其与胃癌发生的关系。HE切片中,根据肠化生占粘膜组织的比例,将其分为Ⅰ、Ⅱ、Ⅲ级;在HID、AB、pH 2.5切片中,根据占优势的粘蛋白,将其分为大肠化生为主型(硫粘蛋白—棕黑色>唾液酸枯蛋白—蓝包)、小肠化生为主型(唾液酸枯蛋白>硫酸粘蛋白)、大小肠化生为主型(硫粘蛋白=唾液酸粘蛋白);将粘液癌分为大肠为主型、小肠为主型、大小肠     

双染技术在腺鳞癌鉴别诊断中的应用

近年来,应用免疫酶组化双标法在同一切片上显示两种或两种以上抗原的新技术报道甚多。本文介绍一种在同一切片上应用免疫组化中PAP法显示腺鳞癌组织中鳞癌细胞内细胞角蛋白抗原;和应用组织化学AB法(Alcain blue),粘液卡红法,PAS反应及AB—PAS染色显示腺     

正常胃粘膜屏障血型抗原的表达及其生物学意义

首次应用国产单克隆抗体、ABC法,检测国人胎儿、成人正常胃组织石蜡切片A、B、H、M和N血型抗原(BGA)的定位,结合粘液组化AB(pH 2.5)/PAS染色,判断粘液分泌性质。结果表明:胎儿胃粘膜表面和小凹上皮大部分由粘液细胞组成,BGA表达阳性,分泌混合型粘液;成人胃粘膜表面和小凹上皮完全由粘液细胞组成,BGA表达可分为分泌性(A、B、H)和非分泌性(M、N、A、B)抗原;胃粘液中分泌性抗原主要由表面和小凹上皮粘液细胞分泌。提出BGA为胃粘膜屏障的重要组成,为进一步研究胃疾病提供正常形态学和免疫遗传学对照资料。     

乳腺粘液腺癌组织学、组织化学研究

对20例乳腺粘液腺癌进行了组织学、组织化学研究。根据肿瘤是否伴有它型乳癌,将之分为纯粘液腺癌及混合型粘液腺癌二类。组化染色显示粘液属酸性粘液中的唾液酸粘液。并探讨了肿瘤的组织起源。     

乳腺分泌性癌的临床病理和免疫组织化学分析

目的 探讨乳腺分泌性癌的临床病理特征、免疫组织化学表达和特殊染色特点及其生物学意义。方法 分析3例乳腺分泌性癌的临床表现和病理特征;用黏液卡红（MC）和PAS/AB（Alcian blue）及淀粉酶消化后PAS/AB染色观察分泌物的性质,同时采用免疫组织化学ABC法检测ER、PR、Ki-67、p53、p63、c-erbB-2、S-100蛋白、CK7、SMA、CK1/3、EMA、CEA和CK34βE12在肿瘤细胞中的表达。结果 乳腺分泌性癌仅占同期全部活检乳腺癌的0.36%,平均年龄48.7岁。病理形态学的主要特点为癌细胞异型性较小,出现微囊性结构,癌细胞能产生大量分泌物等,对PAS/AB及黏液卡红染色呈现阳性反应。某些病例可以出现印戒样细胞,柱状上皮被覆的腺样结构,及间质广泛透明变性,癌细胞萎缩消失后形成的胶原硬化瘢痕,提示其高分泌活性和相对惰性的生长方式。尽管ER、PR阴性或仅局灶性表达,但预后相关基因p53和c-erbB-2全阴性或仅偶见弱表达,另外,Ki-67反映的增殖指数仅为6.3%,可能同其相对良好的组织形态和较好的预后有关。淋巴结均未见转移。结论 乳腺分泌性癌为一种少见的肿瘤,可发生于中老年女性,侵袭性和转移能力弱,预后良好。根据组织学表现,结合特染及免疫组化表达特点,可作出明确的病理诊断。     

Intramucosal cysts of the stomach. VIII: Histochemical studies

Intramucosal cysts of the human stomach have been earlier classified on the basis of their epithelial lining into 1. fundic, 2. foveolar, 3. pyloric, 4. with intestinal metaplasia and 5. with ciliated metaplasia. Four histochemical methods (high iron diamine (HID)-Alcian blue pH 2.5 (AB), PAS, Concanavalin A (ConA), and Grimelius reaction were used. The cells of foveolar cysts contained neutral mucins and sialomucins, and those of pyloric cysts, neutral mucins, sialomucins, mannose-rich glycoprotein and argyrophilic material. The goblet cells in intestinal metaplastic cysts contained neutral mucins and sialomucins as well as sulphated mucins, while ciliated cells in ciliated metaplastic cysts demonstrated mannose-rich glycoproteins and argyrophilic material (although some ciliated cells were negative for both). The cells of fundic gland cysts were negative for all tested methods. The frequency of intramucosal gastric cysts is known to be high in stomachs having adenocarcinoma, and low in stomachs with peptic ulcers. Several reports have demonstrated alterations in the composition of gastric mucins in stomachs harbouring an adenocarcinoma. Consequently, the histochemical stains may prove of value to investigate the true significance of intramucosal cysts in gastric carcinogenesis.     

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.     

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.     

A study of vesical adenocarcinoma, intestinal metaplasia and related lesions using mucin histochemistry

Biopsy and autopsy material from the urinary bladder was studied using PAS and PAS-D techniques to identify glycogen and neutral mucins, the alcian blue/high iron diamine method to distinguish sialo- and sulphamucins and the PB/KOH/PAS technique to localize O-acylated sialomucins. All of 10 examples of normal urothelium and both of two cases of transitional carcinoma in situ contained glycogen, but no mucin. Other lesions displayed one of two patterns of mucin production: the extracellular mucin seen focally in 17 cases of cystitis cystica consisted of sialo- and/or neutral mucins only, a pattern also displayed by mucins produced in 10 of 13 examples of transitional cell carcinomas and by three of nine tumours purely or in part adenocarcinomas. The intracellular mucins expressed in five of the 17 cases of cystitis glandularis and in all of eight cases of frank intestinal metaplasia with goblet cells displayed a colonic phenotype, with production of O-acylated sialomucins. A similar profile was expressed by six adenocarcinomas and this included tumours likely to be of vesical and also of urachal origin. It is concluded that identification of O-acylated mucins cannot distinguish between primary bladder tumours and metastases from a colonic primary, or between carcinomas of vesical and urachal origin.     

Mucin histochemistry of heterotopic gastric mucosa of the upper esophagus in adults: possible pathogenic implications

The mucin profile of 24 endoscopic biopsies of heterotopic gastric mucosa (HGM) of the upper esophagus in adults and a control group of ten cases of Meckel's diverticula containing heterotopic gastric mucosa were studied with two combined histochemical methods: alcian blue pH 2.5/PAS and high iron diamine/alcian blue pH 2.5. The clinical and light microscopic features of the 24 HGM cases were also reviewed. In addition to overall secretion of neutral mucins by the 24 HGM cases, mucin histochemistry showed prominent secretion of acidic mucins in 19 of 24 HGM cases (79%), with sulphomucins in 11 of 24 HGM cases (45.8%). This mucin profile of HGM was unlike that of either normal gastric mucosa or heterotopic gastric mucosa in Meckel's diverticula. Moreover, a comparison between the mucin profile and clinical features of HGM and Barrett's esophagus showed certain similarities. The data suggest a physiopathologic link between HGM and Barrett's esophagus.     

Mucins in Barrett's esophagus: a histochemical study

To define the mucin secretion of Barrett's esophagus, 132 biopsy specimens from 37 patients were studied with histochemical stains for mucins. Columnar mucous cells contained largely neutral mucins, but scattered cells in over 70% of the biopsies also produced sialomucins and sulfomucins. Goblet cells contained sialomucins, sulfomucins, or both. The distribution and relative proportions of mucins varied considerably among biopsies and among patients. Barrett's esophagus histochemically represents a heterogeneous, partially differentiated epithelium analogous to incomplete intestinal metaplasia of gastric mucosa.     

Simple procedure for assessing relative quantities of neutral and acidic sugars in mucin glycoproteins: its use in assessing cyclical changes in cervical mucins.

A simple histochemical procedure for assessing relative amounts of neutral and acidic sugars in mucin glycoproteins, and its application in the study of cyclical changes of human cervical mucins, is described. This procedure, the saponification/selective periodate oxidation/borohydride reduction/alcian blue pH 2.5/periodic acid Schiff (KOH/PA*/Bh/Ab 2.5/PAS) method, uses a selective oxidation step to remove the PAS positivity of sialic acid; thus only neutral sugars stain positively with PAS, and acidic sugars (O-sulphate esters and carboxyl groups) stain with alcian blue. This differs from the KOH/Ab/PAS technique which stains sialic acid residues with both alcian blue and PAS. Applying the KOH/PA*/Bh/Ab 2.5/PAS technique to the study of cyclical changes of human cervical mucins, a decreased neutral:acidic sugar ratio in the secretory phase mucins compared with those of the proliferative phase was found. This difference was not seen with KOH/Ab/PAS staining in the same cases. The techniques and reagents used in this procedure can be easily applied in a clinical histopathology laboratory.     

Histochemistry of mucosubstances in the frog respiratory epithelium

The histochemical nature of the mucosubstances of the respiratory epithelium of the frog palate was studied. 3 main types of epithelial cells were identified: Goblet cells which contained some glycogen, some neutral mucins and a mixture of sialidase-resistant and sialidase-labile sialomucins, non ciliated columnar cells with a secretory activity restricted to their apical pole. These cells contained abundant glycogen, some neutral mucins and sialidase-resistant sialomucins, ciliated cells with no secretory activity. The surface mucous blanket was composed of a mixture of predominant sialomucins and some neutral mucins. The histochemistry of mucosubstances in the respiratory epithelium of the frog palate differed from that of human and most respiratory systems by the presence of glycogen and the absence of sulphated mucins.     

Immunohistochemical detection of O-acetylated sialomucins in intestinal metaplasia and carcinoma of the stomach

Eighty-two selected gastric mucosal biopsy or resection specimens were stained both conventionally, to classify subtypes of intestinal metaplasia and carcinoma, and immunohistochemically with a mouse monoclonal antibody (MMM-17), raised against normal human colonic mucin, which has an affinity for di- and/or tri-O-acetylated sialomucin. The aims of the study were to reassess the prevalence of O-acetylated sialomucins in normal, metaplastic and carcinomatous gastric mucosa and to investigate whether the production of these mucins by intestinal metaplasia is related to its associated mucosal pathology. O-acetylated sialomucins were not seen in normal mucosa. They were, however, prevalent in all sub-types of metaplastic (64.8%) and carcinomatous (42.9%) mucosa. Type 1 intestinal metaplasia was significantly more likely to contain this type of mucin if Helicobacter pylori infection was identifiable in the adjacent gastric mucosa (81.0% v. 38.5%, P < 0.025). Type 3 showed a similar, albeit nonsignificant, relationship (100% v. 62.5%). O-acetylated sialomucins are, therefore, much more prevalent in gastric intestinal metaplasia and carcinoma than previously recognized by conventional staining techniques. The production of this type of mucin by intestinal metaplasia may reflect an adaptive response to alterations in the luminal environment such as an increase in bacterial content.     

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.