Expression of basic fibroblast growth factor in human gastric carcinomas.

The expression of mRNA for the basic fibroblast growth factor (FGF) gene was examined in seven human gastric carcinoma cell lines and in tissue from 29 gastric carcinomas together with the adjacent normal mucosa. Among the seven gastric carcinoma cell lines, the MKN45 cell line expressed mRNA for the basic FGF gene. Basic FGF protein production was confirmed by flow cytometric analysis and immunohistochemistry. Among the surgical specimens, 16 (55%) of 29 gastric carcinomas showed higher levels of basic FGF mRNA than the normal mucosa. Interestingly, in scirrhous gastric carcinomas characterized by their fibrous stroma and rapid growth, 9 (69%) of 13, samples examined revealed higher levels of basic FGF mRNA than normal mucosa, whereas only 3 (33%) of the 9 well differentiated adenocarcinomas studied produced similar results. Immunohistochemically, basic FGF protein was localized in tumor cells. These results suggest that basic FGF produced by tumor cells may play an important role in producing fibrosis and angiogenesis in gastric carcinomas.     

Expression of basic fibroblast growth factor in human gastric carcinomas

The expression of mRNA for the basic fibroblast growth factor (FGF) gene was examined in seven human gastric carcinoma cell lines and in tissue from 29 gastric carcinomas together with the adjacent normal mucosa. Among the seven gastric carcinoma cell lines, the MKN45 cell line expressed mRNA for the basic FGF gene. Basic FGF protein production was confirmed by flow cytometric analysis and immunohistochemistry. Among the surgical specimens, 16 (55%) of 29 gastric carcinomas showed higher levels of basic FGF mRNA than the normal mucosa. Interestingly, in scirr-hous gastric carcinomas characterized by their fibrous stroma and rapid growth, 9 (69%) of 13, samples examined revealed higher levels of basic FGF mRNA than normal mucosa, whereas only 3 (33%) of the 9 well differentiated adenocarcinomas studied produced similar results. Immunohistochemically, basic FGF protein was localized in tumor cells. These results suggest that basic FGF produced by tumor cells may play an important role in producing fibrosis and angiogenesis in gastric carcinomas.     

Ultrastructural localization of vesicle-associated membrane protein(s) to specialized membrane structures in human pericytes, vascular smooth muscle cells, endothelial cells, neutrophils, and eosinophils.

Vesicle-associated membrane proteins (VAMPs) are important to the trafficking of vesicles between membrane-bound intracytoplasmic organelles, in the facilitation of neurosecretion, and in constitutive and regulated secretion in non-neuronal cells. We used a pre-embedding ultrastructural immunonanogold method to localize VAMPs to subcellular sites in human cells of five lineages known to have cytoplasmic vesicles that may function in vesicular transport. We found VAMPs localized to caveolae in pericytes, vascular smooth muscle cells, and endothelial cells of venules, to the vesiculo-vacuolar organelle, recently defined in venular endothelial cells, to the vesicle-rich intergranular cytoplasm and secretory granule membranes of neutrophils, and to perigranular cytoplasmic secretory vesicles and secretory granule membranes in eosinophils. These specific localizations in five human vascular and granulocyte lineages support the notion that VAMPs have vesicle-associated functions in these cells.     

Immunohistochemical detection of metallothionein in carcinomatous and normal human gastric mucosa

Utilising a specific monoclonal mouse antibody (E9), metallothionein (MT) expression has been immunohistochemically investigated in 112 formalin-fixed paraffin-embedded surgical gastric samples, 38 of which were early carcinomas (EGC) and 74 advanced ones (AGC); clinico-pathological details and follow-up data (ranging from 3 to 197 months, mean 60.5 months) were available. Eighty-nine portions of gastric mucosa adjacent to examined carcinomas (transitional mucosa) were also analysed; in addition, 22 biopsies of normal gastric mucosa were studied as tissue control. The MT immunoreactivity was evaluated by staining and intensity-distribution scores. A various MT positivity was appreciable in the cytoplasm and nucleus of antrum or body gastric epithelial cells in 100% of normal control biopsies. 75/112 (67%) gastric carcinomas showed MT immunoreactivity with a significant lower expression in AGC. No relationships were encountered between MT immunostaining and clinico-pathological data; in addition, no difference in the Kaplan-Meier survival curves of patients with various MT expression was achieved. When the transitional mucosa was examined, 84/89 (94%) samples were stained although the immunoreaction was not always concordant with that encountered in adjacent carcinomatous elements. The significant statistical decrease of MT scores observed by us moving from normal to neoplastic gastric mucosa allows us to exclude the hypothesis of an overexpression of MT in gastric carcinomas.     

Endothelial stomatal and fenestral diaphragms in normal vessels and angiogenesis

Vascular endothelium lines the entire cardiovascular system where performs a series of vital functions including the control of microvascular permeability, coagulation inflammation, vascular tone as well as the formation of new vessels via vasculogenesis and angiogenesis in normal and disease states. Normal endothelium consists of heterogeneous populations of cells differentiated according to the vascular bed and segment of the vascular tree where they occur. One of the cardinal features is the expression of specific subcellular structures such as plas-malemmal vesicles or caveolae, transendothelial channels, vesiculo-vacuolar organelles, endothelial pockets and fenestrae, whose presence define several endothelial morphological types. A less explored observation is the differential expression of such structures in diverse settings of angiogenesis. This review will focus on the latest developments on the components, structure and function of these specific endothelial structures in normal endothelium as well as in diverse settings of angiogenesis.     

胃癌神经内分泌细胞免疫细胞化学的研究

９６例胃癌应用铬粒素抗体免疫细胞化学法发现３８例（３９．６％）肿瘤含有神经内分泌细胞（ＮＥ细胞）;呈激素阳性者：血清素５例,生长抑素５例,胰多肽５例,胰高血糖素５例,胃泌素３例,ＡＣＴＨ６例。结果显示：ＮＥ细胞发现率及细胞数量在低分化癌中明显高于高、中分化癌;异位激素──ＡＣＴＨ阳性细胞数量在低分化癌也明显多于高、中分化癌;本位激素阳性细胞在ＮＥ细胞中的比例在非肿瘤性粘膜中明显高于癌组织,在原发肿瘤明显高于转移灶,在正常粘膜又明显高于癌旁粘膜。根据上述结果,对ＮＥ细胞的发现率、方法学及其激素产物进行了讨论。     

Transcription factor snail1 expression and poor survival in pharyngeal squamous cell carcinoma

Snail1, a key regulator of epithelial-mesenchymal transition (EMT), plays an important role in tumour progression. Previous studies of snail1 have mainly focused on the epithelial tumour cells. The objective of this study was to evaluate the expression of snail1 protein in endothelial cells, stromal myofibroblasts and malignant epithelial cells of pharyngeal squamous cell carcinomas (PSCC), as well as its relation to clinicopathological features and survival. One hundred and ten tissue microarray samples were analyzed for snail1 expression using immunohistochemistry. In endothelial cells snail1 expression was observed in 51 (48%) of 107 cases and it predicted reduced disease specific survival (DSS) (p=0.009). In 49 (46%) tumour samples snail1 immunostaining was detected in stromal myofibroblasts and there was a tendency to poorer DSS in that group (p=0.067). Snail1 expression in endothelial cells and stromal myofibroblasts is also associated with hypopharyngeal tumours (p=0.01 and p=0.038 respectively), increasing T category (T3-4) (p=0.005, p=0.037 respectively) and poorer general condition of the patient (Karnofsky performance status score <70; p=0.029, p=0.039 respectively). Moreover endothelial expression correlated with advanced stage (III-IV) (p=0.005) and poorer differentiation (grade 2-3; p=0.012). In malignant epithelial cells snail1 immunostaining was detected in 75 of 110 cases (68%). Expression of the protein was more common in hypopharyngeal tumours (p=0.044). Snail1 positive tumours associated with a lower Karnofsky performance status score (p=0.039) and regional failure (p=0.042). Our findings indicate that snail1 protein expression in endothelial cells and to some extent also in tumour stromal myofibroblasts seems to be a predictor of poor survival in PSCC. The presence of snail1 protein in tumour microenvironment rather than in malignant epithelial tumour cells may induce tissue remodelling and tumour progression.     

Immunohistochemical study of the expression of MUC6 mucin and co-expression of other secreted mucins (MUC5AC and MUC2) in human gastric carcinomas.

To investigate the expression of MUC6 mucin in gastric carcinomas, we generated a novel monoclonal antibody (MAb CLH5) using an MUC6 synthetic peptide. MAb CLH5 reacted exclusively with the MUC6 peptide and with native and deglycosylated mucin extracts from gastric tissues. MAb CLH5 immunoreactivity was observed in normal gastric mucosa restricted to pyloric glands of the antrum and mucopeptic cells of the neck zone of the body region. In a series of 104 gastric carcinomas, 31 (29.8%) were immunoreactive for MUC6. The expression of MUC6 was not associated with histomorphological type or with clinicopathological features of the carcinomas. Analysis of the co-expression of MUC6 with other secreted mucins (MUC5AC and MUC2) in 20 gastric carcinomas revealed that different mucin core proteins are co-expressed in 55% of the cases. MUC6 was co-expressed and co-localized with MUC5AC in 45% and with MUC2 in 5% of the cases. Expression of MUC2 alone was observed in 25% of the cases. All carcinomas expressing MUC2 mucin in more than 50% of the cells were of the mucinous type according to the WHO classification. The co-expression of mucins was independent of the histomorphological type and stage of the tumors. In conclusion, we observed, using a novel well-characterized MAb, that MUC6 is a good marker of mucopeptic cell differentiation and is expressed in 30% of gastric carcinomas, independent of the clinicopathological features of the cases. Furthermore, we found that co-expression and co-localization of mucins in gastric carcinomas is independent of histomorphology and staging. Finally, we observed that intestinal mucin MUC2 is expressed as the most prominent mucin of the mucins tested in mucinous-type gastric carcinomas.     

Endocrine cells in gastric carcinoma and adjacent mucosa. An immunohistochemical and ultrastructural study

Endocrine cells are often found in human gastric carcinoma and may be recognized by the immunoreactivity of their chromogranin A, peptides and biogenic amines content. Anti-chromogranin A was used to investigate the morphology of endocrine cells using light and electron microscope immunohistochemical techniques. The hormone content of endocrine cells was examined in both tumour tissue and tumour-adjacent mucosa. It was found that the endocrine cells in tumour tissue were malignant, often had amphocrine differentiation and did not resemble a normal cell type. The hormone content of endocrine cells in tumour tissue seldom corresponded to the hormonal content of endocrine cells in tumour-adjacent mucosa. In intestinal-type carcinoma and in some parts of diffuse-type gastric carcinomas, endocrine cell hyperplasia and an alteration of the differentiation in the tumour-adjacent mucosa were discovered. The distribution of endocrine cells in the tumour tissue was different in both types of gastric carcinoma. The results reported here suggest that endocrine cell differentiation of malignant endocrine cells in human gastric carcinoma develops in a different way from that of endocrine cells in tumour-adjacent mucosa, and as a result, diverse hormonal products may appear in tumour tissue.     

Molecular pathogenesis of hepatic fibrosis and current therapeutic approaches

The pathogenesis of hepatic fibrosis involves significant deposition of fibrilar collagen and other extracellular matrix proteins. It is a rather dynamic process of wound healing in response to a variety of persistent liver injury caused by factors such as ethanol intake, viral infection, drugs, toxins, cholestasis, and metabolic disorders. Liver fibrosis distorts the hepatic architecture, decreases the number of endothelial cell fenestrations and causes portal hypertension. Key events are the activation and transformation of quiescent hepatic stellate cells into myofibroblast-like cells with the subsequent up-regulation of proteins such as α-smooth muscle actin, interstitial collagens, matrix metalloproteinases, tissue inhibitor of metalloproteinases, and proteoglycans. Oxidative stress is a major contributing factor to the onset of liver fibrosis and it is typically associated with a decrease in the antioxidant defense. Currently, there is no effective therapy for advanced liver fibrosis. In its early stages, liver fibrosis is reversible upon cessation of the causative agent. In this review, we discuss some aspects on the etiology of liver fibrosis, the cells involved, the molecular pathogenesis, and the current therapeutic approaches.     

Distribution of the carcinoembryonic antigen CEA in gastric lesions. Immunohistochemical testing of three novel monoclonal antibodies

Three mouse monoclonal antibodies MAB (CEA 12-140-1, -2 and -4) raised against different CEA epitopes were tested in 32 gastric adenocarcinomas (18 intestinal type and 14 diffuse type) and 34 gastric lesions with severe and moderate dysplasia. The MAB stained 13, 11 and 13 out of the 14 diffuse carcinomas and 11, 13 and 13 out of the 18 intestinal carcinomas. The dysplastic lesions were positive in 9, 9 and 6 out of 34 cases. Less than half of the cases with metaplastic epithelium adjacent to the carcinomas were also positive for MAB. All MAB showed the same pattern of reactivity without cross-reactivity. Their cumulative staining rate corresponded closely to that of polyclonal CEA antiserum, but the MAB stained more cells. The reactivity was confined to intracytoplasmic vacuoles in diffuse carcinomas and appeared diffusely in the cytoplasm or limited to the cell membrane in intestinal type of carcinomas. Our findings do not indicate CEA to be a reliable marker for malignant transformation in gastric mucosa.     

Patterns of regeneration in vessels of human diseased muscle and skin

Regenerating vessels from 36 muscle biopsies and 12 skin biopsies pertaining to patients with the clinical and histological diagnosis of dermatomyositis-polymyositis and other inflammatory myopathies were described ultrastructurally. The following characteristics of vascular regeneration were encountered: a) alternation of thin and thick endothelial cells, b) superimposed segments of endothelial cytoplasm with formation of twisted intercellular junctions, c) long and complex intraluminal endothelial projections, d) discontinuous and porous perivascular basement lamina or multilaminated basement lamina. Some vessels featured only two or three of the mentioned characteristics. Endothelial fenestrations were occasionally encountered in the skin capillaries and were an exception in the muscle capillaries. In the skin there were aspects suggesting that intercalation of perivascular cells in the regenerating endothelial wall may occur. Most capillaries were in the final stage of regeneration.     

Flow cytometric analysis of paraffin-embedded material in human gastric cancer

Flow cytometric DNa analysis was performed on formalin-fixed, paraffin-embedded samples obtained by gastroscopic biopsy from 9 patients with histologically normal gastric mucosa (36 specimens) and by radical gastrectomy from 42 cases of human gastric cancer (120 specimens). Ploidy patterns and the distribution of cells in the different cell cycle phases were estimated, and the results were correlated with the histologic and clinical features. All samples of normal mucosa showed a diploid modal DNA content whereas DNA aneuploidy was encountered in 71.4% of the gastric tumors. The correlation between aneuploidy and histologic malignancy grading was statistically significant: aneuploidy was found in 36.4% of highly differentiated (grade 1 and grade 2) tumors and in 75.0% of poorly differentiated (grade 3) tumors (P less than .05). The percentage of cells in S-phase in normal gastric mucosa (median: 5.0%) was lower than that in the tumors (median: 11.3%) (P less than .05). There was a trend for grade 3 tumors to have higher median values (median: 13.4%) than grade 1 and 2 tumors (median: 9.3%); however, this was not statistically significant. An aneuploid DNA pattern was associated with a poorer prognosis, both in early and in advanced stages of gastric tumors, while proliferative activity did not correlate with postoperative survival.     

Ghrelin expression in gut endocrine growths

We aimed to assess the occurrence of ghrelin, a new gut hormone, in endocrine growths of the stomach. In addition, since ghrelin has been detected in other gut derivatives during adult and/or fetal life, we also studied endocrine tumours of the pancreas, intestine and lung. A specific serum generated against amino acids 13-28 of ghrelin was tested on 16 specimens of gastric mucosa with endocrine cell hyperplasia and on 75 endocrine tumours. Ghrelin-immunoreactive cells were moderately represented in normal, atrophic or hypertrophic gastric mucosa, as a rule with no obvious hyperplastic changes even in the presence of concurrent, prominent enterochromaffin-like cell hyperplasia associated with hypergastrinemia. Ghrelin cells were also found in tumour cell fractions of well-differentiated gastric (25 of 33, 76%), pancreatic (6 of 15, 40%) and pulmonary (4 of 8) endocrine tumours. No ghrelin immunoreactivity was detected in 14 intestinal tumours and in five poorly differentiated endocrine carcinomas of the stomach or pancreas. We conclude that ghrelin cells may take part in gut endocrine growths, with special reference to well-differentiated endocrine tumours of the stomach, independently from associated signs of endocrine hyperfunction.     

The vesiculo-vacuolar organelle (VVO). A new endothelial cell permeability organelle.

A newly defined endothelial cell permeability structure, termed the vesiculo-vacuolar organelle (VVO), has been identified in the microvasculature that accompanies tumors, in venules associated with allergic inflammation, and in the endothelia of normal venules. This organelle provides the major route of extravasation of macromolecules at sites of increased vascular permeability induced by vascular permeability factor/vascular endothelial growth factor (VPF/VEGF), serotonin, and histamine in animal models. Continuity of these large sessile structures between the vascular lumen and the extracellular space has been demonstrated in kinetic studies with ultrastructural electron-dense tracers, by direct observation of tilted electron micrographs, and by ultrathin serial sections with three-dimensional computer reconstructions. Ultrastructural enzyme-affinity cytochemical and immunocytochemical studies have identified histamine and VPF/VEGF bound to VVOs in vivo in animal models in which these mediators of permeability are released from mast cells and tumor cells, respectively. The high-affinity receptor for VPF/VEGF, VEGFR-2, was localized to VVOs and their substructural components by pre-embedding ultrastructural immunonanogold and immunoperoxidase techniques. Similar methods were used to localize caveolin and vesicle-associated membrane protein (VAMP) to VVOs and caveolae, indicating a possible commonality of formation and function of VVOs to caveolae.     

Ultrastructural identification of umbilical cord vein endothelium in situ and in culture

The ultrastructure of human umbilical cord vein endothelium in situ, after isolation by collagenase treatment, and in primary culture is described. The cultured cells formed a monolayer with typical "butt" and interdigitated junctions with specialized areas, and contained Weibel-Palade bodies, rod-shaped tubular organelles considered specific of endothelial cells. These morphological features were not present in cultures of human skin fibroblasts and fibroblast-like cells derived from umbilical cords. It is thus concluded that endothelial cells retain their characteristic fine structure in primary culture. Simple ultrastructural studies can thus be used to identify endothelial cells in culture.     

Scrotal angiokeratoma (Fordyce): histopathological and ultrastructural findings.

Bioptic findings related to four cases of scrotal angiokeratoma-Fordyce, were studied under light and electron microscopy. A particular heterogeneity of the structural and ultrastructural patterns typical of this lesion was thus observed. Light microscopy study pointed out, in particular, different degrees of dilation of papillary vessels, whereas ultrastructural study highlighted marked alterations of endothelial cells with structural and quantitative modifications of cytoplasmic organelles.     

LMP-1在胃癌和相应癌旁组织中的表达

目的探讨Epstein-Barr virus(EBV)感染与胃癌发生发展的关系。方法应用免疫组化SP法检测EB病毒潜伏感染膜蛋白-1(LMP-1)在97例胃癌组织及89例相应癌旁组织中表达。结果97例胃癌组织中30例LMP-1蛋白表达阳性,阳性率为30.9%,EBV阳性率与患者性别、浸润深度、淋巴结转移、组织学分型和临床分期之间无明显关系(P>0.05);89例相应癌旁组织中33例检测到EBV LMP-1的表达,胃癌组织及相应癌旁组织EBV阳性表达之间有显著相关性(P=0.000)。结论EBV感染与胃癌的发生有一定的相关性。     

Ultrastructure of cells of proper gastric glands and their stroma in hemorrhagic shock in the rat.

Ultrastructural changes in the cells of the proper gastric glands and their stroma are biphasic in the hemorrhagic shock. First phase: "paralysis" of the capillary vessels with an oedema of their stroma, an intracellular oedema with hydropic degeneration of the parietal cells and a degranulation of the argentaffin and argentaffin-like cells. Second phase: ischemia of the gastric mucosa with ultrastructural features of: a) the increased secretion of the parietal cells, b) the degranulation of the chief gastric cells, c) the increased secretion processes in the mucous neck cells. These findings suggest that at last 2 factors coincide in the pathogenesis of so-called stress gastric ulcerations: biphasic disturbances of blood circulation initially damage the gastric mucosa and are followed the digestive hyperactivity of the gastric juice finally resulting stress ulceration.