Serotonin in fore-gut carcinoids. A survey of 60 cases with regard to silver stains, formalin-induced fluorescence and serotonin immunocytochemistry

A series of 60 fore-gut carcinoid tumours was examined with regard to serotonin content after application of three different techniques, namely: the argentaffin reaction, formalin-induced fluorescence according to Falck-Hillarp and immunocytochemistry with monoclonal antibodies to serotonin. To evaluate the staining-fluorescence of individual tumour cells, the methods were applied to identical tumour sections. Twelve tumours demonstrated serotonin-immunoreactive cells, six of which were also argentaffin. Four tumours contained argentaffin cells but no serotonin-immunoreactivity. With the use of all three techniques, three types of tumour cells occurred, namely: serotonin-immunoreactive, non-argentaffin and non-fluorescence cells, serotonin-immunoreactive, argentaffin and fluorescent cells, and non-serotonin immunoreactive, argentaffin and non-fluorescent cells. The first (serotonin-immunoreactive) cell type was most frequently found in the tumours. One gastric carcinoid in which the argentaffin cells exceeded the serotonin-immunoreactive cells, a positive reaction was found with the modified Warthin-Starry reaction for demonstrating melanin. Since none of the techniques used for visualization of serotonin in endocrine tumours is unquestionably specific and since they do not give identical results, it is indicated that for a more accurate identification of serotonin in fore-gut carcinoid tumours, a positive reaction with at least two of the applied techniques is desirable.     

Immunocytochemical localization of prohormone convertase 1/3 and 2 in gastrointestinal carcinoids

Gastrointestinal carcinoids are derived from the diffuse intestinal endocrine system and may produce amines and many peptides, including serotonin, chromogranin A (CGA), and tachykinins. Most peptide hormones are synthesized as bigger prohormones, which are processed to smaller active hormones by prohormone convertases (PCs). A total of 35 cases of gastrointestinal carcinoids, including gastric, duodenal, small intestinal, appendiceal, and large intestinal carcinoids, were immunocytochemically stained for serotonin, CGA, and PC 1/3 and 2, in order to colocalize CGA and PCs in the carcinoids. All carcinoids were positive for CGA and PCs. Carcinoids that stained strongly for CGA were generally weakly stained for PCs and those weakly staining for CGA were more strongly stained for PCs in the majority of the small and large intestinal tumors. Gastrointestinal carcinoids were positive for CGA and PCs, and the presence of PCs may suggest that the conversion of peptide prohormones to smaller peptide hormones occurs in gastrointestinal carcinoids. PCs immunocytochemistry may be added as a new phenotypic characterization for gastrointestinal carcinoids.     

Histopathology of gastric carcinoids: a survey of 42 cases

An unselected series of 42 gastric carcinoids has been reviewed. Clinically the tumours simulated common gastric lesions including ulcer, polyp and carcinoma. No endocrine symptoms were identified. The tumours were most frequent in the body of the stomach and in 25% in that site were multiple. Morphologically most tumours when classified according to Soga (1974) demonstrated a mixed growth pattern. Six tumours displayed an atypical morphology (type D): they were larger and metastasized more frequently than the rest of the tumours. Six tumours contained a few scattered argentaffinic cells but the others were negative indicating negligible serotonin secretion in only a few cases. The Grimelius argyrophilic reaction was positive in most cells in all tested tumours except in three, two of which showed atypical morphology (type D). It is suggested that gastric carcinoids with a type D morphology or a minority cell population of argyrophil cells are dedifferentiated carcinoids which are biologically nearer to gastric carcinomas. The most frequent clinicopathological correlation was achlorhydria linking pernicious anaemia and gastric carcinoids. This indicates pathogenetic similarities between gastric carcinoids and gastric carcinomas.     

Chromogranin A and B and secretogranin II in bronchial and intestinal carcinoids

Summary Carcinoid tumours (bronchial and intestinal) were analyzed by immunoblotting for the presence of chromogranin A, B and secretogranin II. In all tumours an antigen corresponding in electrophoretic behaviour to adrenal chromogranin A was present. Lung carcinoids (3 out of 5) contained a relatively high concentration of a proteoglycan form of this antigen in addition. Chromogranin B was found in all tumours. In one and two dimensional immunoblotting it appeared identical to the corresponding adrenal antigen. Secretogranin II was also present, however concentrations (especially in intestinal carcinoids) were low and variable. Furthermore, in intestinal tumours it differed from the adrenal antigen by having a slightly higher molecular size and a more alkaline pI. Immunohistochemistry revealed that the tumour tissues stained positively for all three antigens. For secretogranin II the staining in intestinal tumours was relatively weak and quite variable. These results should provide a defined basis for immunohistochemical screening of carcinoids for the chromogranin/secretogranin antigens.     

原位检测新基因SNC66在胃癌组织与正常胃粘膜中的表达

目的: 观察新基因SNC66在胃癌组织中的表达水平,分析SNC66表达水平与胃癌发生的相关性。 方法:以β-actin为对照,对20例胃癌组织及其配对正常胃粘膜的石蜡切片,进行原位cRNA/mRNA分子杂交和原位RT-PCR扩增。 结果:两种检测方法表明,SNC66在正常胃粘膜组织中都呈强阳性表达。而在胃癌组织中,原位分子杂交检测表明,20例标本SNC66不表达和弱阳性表达各8例,强阳性表达4例。原位RT-PCR检测表明,当循环数为10时,此20例胃癌标本中不表达、弱阳性和呈强阳性表达数分别为6,9和5;而当循环数为25时,则4例不表达,16例呈强阳性表达。 结论:SNC66基因在胃癌组织中存在明显的表达降低和表达缺陷。SNC66可以作为一个胃癌负相关基因加以进一步研究。     

Active neovascularization and possible vascular-centric development of gastric and periscapular elastofibromas

An elastofibroma is a benign and rare fibrous lesion that most commonly occurs in the periscapular region. A gastrointestinal elastofibroma is extremely rare. In the present study, six cases of elastofibromas including a case in the stomach were evaluated. The gastric case revealed widely distributed lesions in the submucosal layer with perivascular fibrotic lesions (PVFLs) and some PVFLs were distributed to the skip lesions of elastofibroma. These PVFLs were also observed in all five periscapular cases and invariably contained elastic fibers which showed various degree of maturation. CD34-positive stromal cells were observed not only in elastofibromas but also in PVFLs in each case. These findings suggested the possibility of the PVFLs were the primary lesions of elastofibroma and their vascular-centric development. The percentage of the CD105-positive vessels in elastofibroma group was significantly higher than in the control group. This result indicates active neovascularization in elastofibromas.     

The importance of smooth muscle cells in the development of foam cells in the gastric mucosa

Summary Foam cells in lipid islands of the stomach can develop from both histiocytes and smooth muscle cells. With increasing storage of lipid vacuoles in smooth muscle cells, loosening of the myofilament arrangement and decrease of the dense areas subjacent to the plasma membrane occurs. Endoplasmic reticulum and the cisternae of the Golgi-apparatus dilate, the cell organelles increase initially and the basement membrane of the smooth muscle cells is fragmentarily formed. Only in incompletely formed foam cells can the origin from smooth muscle cells be recognised, in their final state their histiogenesis is seldom apparent.The authors are grateful to H. Gerdes for leaving the specimens and to R. Naumann and H. Staubitz for technical and photographic assistance.     

Arteriography of three models of gastric oesophagoplasty: the whole stomach, a wide gastric tube and a narrow gastric tube

The esogastric anastomotic fistula,occurring after the replacement of esophagus by the stomach, is a post-operative complication always feared and awaited. Apart from other causes, there exist the anatomical dispositions notably the vascular and technical factors that stress this potential risk despite certain advantages of esophagogastroplasty. The goal of our study was to study the arterial distribution of the gastric transplants in order to identify the better modalities of their making. We used 39 stomachs taken from fresh cadavers of autochtone subjects. After a modeling treatment using three different techniques, they were subjected to a radiographic opacification of the right gastro-epiploic artery with sulphate of barium follow by an x-rays in incidence full-face (25 kv, 10 mAS). It was a matter of 15 entire stomachs (E.E.) with denudation of the small curvature, of 12 wide gastric tubes (W.T.) prepared according to the Akiyama technique modified and of 12 narrow tubes (N.T.) tubulized according to the Marmuse method. We studied the anastomotic type of the gastro-epiploic arterial circle according to the classification of Koskas, the collateral branches of the arterial circles of the gastric curvatures, the antral and corporeal anastomosis of these circles and the distribution anastomotic at the level of the summit of the anastomotic. Only 28 pieces (15 E.E., 8 W.T. and 5 N.T.) were able to be the object of a complete angiographic exploitation. The anastomosis of the arterial circle was type I in 64.1% of the cases, type II in 15.4% of the cases, type III in 15.4% of the cases and type IV in 5.1% of the cases. The average number of collateral branches originating from gastro-epiploic arterial circle was respectively 24, 17 and 22 for the E.E., the W.T. and the N.T. Only the two first ones presented collateral branches being borne of the small curvature circle. Fifty per cent of the N.T. did not possess any antral or corporeal anastomosis between the two arterial circles; some of them were even for a quarter of the W.T. In the case of gastric tubulization there existed an irrigation defect of the summit of the plasty for a third of the N.T. and a quarter of the W.T., despite a constant intramural bridge anastomosis between the two gastro-epiploic arteries. The usage of the entire stomach must be recommended for gastric oesophagoplasty; but when the operative indications require a resection of the small curvature it is preferable to use a wide gastric tube whose diameter respects the two left third of the initial width of the organ.     

B7-H4实时荧光定量PCR检测方法的建立及其在人胃癌组织中的初步应用

目的建立实时荧光定量PCR检测B7-H4的方法并初步应用于人胃癌组织。方法将B7-H4和内参GAPDH的PCR扩增产物经测序鉴定正确后克隆入载体pMD19-T,构建重组质粒标准品,并纯化、定量及梯度稀释,分别建立B7-H4和GAPDH的标准曲线,应用实时荧光定量PCR检测8例人胃癌组织中的B7-H4相对于GAPDH的表达情况。结果 B7-H4的最低检测拷贝数为5.27拷贝,线性范围5.27×101~5.27×107拷贝,标准曲线方程y=–3.1395x+41.805,直线回归相关系数r=0.994904,批间变异系数范围2.39%~3.59%,扩增效率108.2%;GAPDH的最低检测拷贝数为38.6拷贝,线性范围3.86×102~3.86×107拷贝,标准曲线方程y=–3.2436x+41.083,直线回归相关系数r=0.998913,批间变异系数范围2.26%~3.86%,扩增效率103.4%。8例人胃癌组织的B7-H4相对于GAPDHmRNA表达水平在0.044~0.888之间。结论荧光定量PCR检测B7-H4的方法具备较高的敏感性和特异性,且系统有良好的重复性和线性范围。     

胃泌素促进胃癌细胞的迁移和侵袭

 目的：研究胃泌素在胃癌细胞迁移和侵袭中的作用。方法：用细胞划痕实验、Transwell小室迁移和侵袭实验检测10 nmol/L和100 nmol/L胃泌素处理后胃癌细胞AGS和SGC-7901迁移和侵袭能力的变化, MTT法检测细胞增殖能力, ELISA法检测细胞上清液中基质金属蛋白酶2（MMP-2）的含量。结果：10 nmol/L和100 nmol/L胃泌素处理胃癌细胞后,细胞迁移和侵袭能力增强。对照组、10 nmol/L和100 nmol/L胃泌素组AGS细胞平均迁移数分别为56.0、88.1和106.4 /view,SGC-7901细胞平均迁移数分别为52.8、91.0和113.3 /view, AGS细胞平均侵袭数分别为78.4、118.7和141.6 /view,SGC-7901细胞平均侵袭数分别为87.3、124.6和147.4/view（均P<0.01）;100 nmol/L胃泌素组细胞迁移和侵袭能力均高于10 nmol/L组（P<0.05）;胃泌素处理后细胞的增殖率及MMP-2的分泌量也显著增加（P<0.05）。结论：胃泌素通过促进MMP-2的分泌以剂量依赖的方式增强胃癌细胞的迁移和侵袭能力,可能是体内胃癌细胞增殖、侵袭和转移的重要机制之一。     

胃癌组织中Cyr61和核因子-κB的表达与转移及预后的关系

目的 检测胃癌组织中富含半胱氨酸肝素结合蛋白61(Cyr61)和核因子-κB(NF-κB)的表达及其与临床病理特征及预后的关系.方法 应用逆转录聚合酶链反应技术验证和检测53例胃癌组织与11例非肿瘤胃黏膜中Cyr61基因的表达,并用免疫组织化学方法 (SP法)检测99例胃癌组织与25例非肿瘤胃黏膜中cyr61与NF-κB蛋白的表达情况.结果 胃癌原发灶和转移灶中Cyr61mRNA的阳性表达率分别为84.6%(22/26)、88.9%(24/27),明显高于非肿瘤胃黏膜组5/11(P值均＜0.05).非肿瘤胃黏膜、原发癌与转移癌组的基因表达量(2-△Ct)分别为(2.76±5.50)×10-5、(14.61±20.64)×10-5、(18.46±26.38)×10-5.胃癌原发灶、转移灶Cyr61 mRNA均高于非肿瘤胃黏膜(P值均＜0.05),转移灶与原发灶比较,差异无统计学意义(P＞0.05).99例胃癌中Cyr61及NF-κB蛋白的高表达率分别为56.6%(56/99)和55.6%(55/99),其表达水平与肿瘤浸润深度、脉管侵犯、淋巴结转移、远处转移及TNM分期呈正相关(P值均＜0.05),NF-κB还与肿瘤大小呈正相关(P＜0.05);Cyr61蛋白的表达水平与NF-κB呈正相关(P＜0.05);Cyr61与NF-κB蛋白高表达的病例平均生存时间和5年生存率明显低于低表达的病例(P值均＜0.05).结论 Cyr61和NF-κB阳性表达与胃癌的侵袭、转移密切相关,检测Cyr61和NF-κB的表达可作为判断胃癌生物学行为的重要参考指标.     

亚硝基胍诱发大鼠胃腺癌发生发展过程中p53、ras基因表达和突变的研究

目的探讨ｐ５３、ｒａｓ基因在亚硝基胍（ＮｍｅｔｈｙｌＮ′ｎｉｔｒｏＮｎｉｔｒｏｓｏｇｕａｎｉｄｉｎｅ，ＭＮＮＧ）诱发大鼠胃腺癌发生发展过程中的作用。方法用免疫组织化学、聚合酶链反应单链构象多态性分析（ＰＣＲＳＳＣＰ）技术对大鼠正常腺胃粘膜、癌前病变和胃腺癌组织中ｐ５３，ｒａｓ基因的表达和突变进行检测。结果ｐ５３蛋白在癌组织中的阳性表达率为５０％（２０／４０只），正常和癌前病变组织中为阴性；ｒａｓ蛋白（ｐ２１）癌前病变组织中阳性率为４４％（２３／５２只），在癌组织中为２３％（９／４０只）。癌组织中ｐ５３基因突变率为４５％（１８／４０只），非癌组织为阴性。癌组织及癌前病变组织中均未发现ｒａｓ基因突变。结论在ＭＮＮＧ诱发大鼠胃腺癌发生发展过程中，ｒａｓ基因的激活是一早期事件，可能参与了癌的发生过程，而ｐ５３突变则主要与胃癌的进展有关。     

侧脑室注射微量五肽胃泌素对大鼠胃电和胃运动的影响

在大鼠侧脑室微量注入五肽胃泌素(G-5)0.1μg观察其对胃电和胃运动的影响。实验结果表明:侧脑室注射G-5(0.1μg/10μl)后胃电慢波的振幅由对照值的0.56mV增加到0.847mV(P<0.01),快波的振幅和频率也明显增加;胃窦运动频率增加到对照值的41.556±1.01%(P<0.01),振幅增加196.68±2.07%(P<0.001)。这一反应被切断双侧颈迷走神经所阻断。提示脑内的胃泌素可参与对胃电和胃运动的调节,这一作用可能是通过迷走神经来实现的。     

原发性胃癌组织中的端粒及端粒酶表达

目的观察端粒（ＴＲＦ）及端粒酶在胃癌形成及发展中的作用。方法采用ＤＮＡ琼脂糖凝胶杂交技术及端粒重复扩增ＰＣＲ方法检测１７例早期胃癌、８９例进展期胃癌组织及邻近正常组织中的平均ＴＲＦ长度及端粒酶活性。结果早期、进展期胃癌的ＴＲＦ长度及端粒酶活性表达均显著短于或高于邻近胃组织（Ｐ＜０．０５～０．０１），且端粒酶活性的表达主要表现在ＴＲＦ缩短或延长的胃癌组织中，而ＴＲＦ缩短与胃癌的分化程度相一致。结论端粒及端粒酶的异常状态可能与胃癌的发生发展密切相关，端粒酶活性及端粒缩短可以作为胃癌的诊断标志     

Esthesioneuroblastoma

Summary A case of esthesioneuroblastoma was examined by histological, histochemical and electron microscopic techniques. The tumour cells showed an argyrophil reaction with the Grimelius technique and contained cytoplasmic secretory granules, but in contrast to previous reports were devoid of histochemically demonstrable biogenic amines. For routine diagnosis the argyrophil technique may be useful in differentiating this type of tumour from epidermoid carcinoma.Supported by the Swedish Medical Research Council (Project No. 102) and Landsforeningen for kraeftens bekaempelse (2206/76)     

Fetal development of the pyloric muscle

Summary The fetal development of the pyloric muscle was studied in five human embryos (crown-rump length 5 to 31 mm) and in ten fetuses aged 3 to 9 months. Samples of pyloric muscle were obtained during operation for pyloric stenosis in two infants aged six weeks. Anatomo-radiologic, morphologic and immunohistochemical studies were made on this material, from which it emerged that the pylorus is identifiable by means of specific markers from the 40th day. Its two-layered muscular structure is described in detail. The mechanism of sphincteric function is reviewed. This study assumes clicincal importance in the context of the etiopathogenesis of hypertrophic stenosis of the pylorus.Work presented at the First European Congress of the European Association of Clinical Anatomy, September 9–10, 1991, Brussels, Belgium     

胃癌及肠化组织微卫星不稳定性

目的研究微卫星不稳定性（ＭＳＩ）在胃癌发生、发展中的作用。方法采用聚合酶链反应（ＰＣＲ）方法检测了５０例手术切除胃癌标本及１５例肠化标本的ＭＳＩ。结果５０例胃癌中有２７例检出１个以上位点ＭＳＩ,总阳性率为５４０％;高中分化腺癌ＭＳＩ检出率（８６．６％）显著高于低分化腺癌（３８．７％,Ｐ＜０．０５）;肠型胃癌ＭＳＩ阳性率（７７．８％）显著高于胃型胃癌（４００％,Ｐ＜０．０５）;ＭＳＩ与胃癌发病年龄、大小、发生部位、浸润深度、淋巴结转移及临床分期无显著相关。３例早期胃癌ＭＳＩ均阳性,１５例肠化标本中有３例检出ＭＳＩ,阳性率为２０％。结论ＭＳＩ是胃癌发生过程中的早期分子标志,在胃癌的发生中可能扮演重要角色。