Non-random spatial relationships between mast cells and microvessels in human endometrial carcinoma

Mast cells (MCs) accumulate in the stroma surrounding tumors, where they secrete angiogenic cytokines and proteases, and an increased number of MCs have been demonstrated in angiogenesis associated with solid and hematological tumors. The aim of this study is to contribute to the knowledge of distribution of MCs in tumors, investigating the pattern of distribution of tryptase-positive MCs around the blood vessels in human endometrial carcinoma samples by introducing a quantitative approach to characterize their spatial distribution. The results have shown that in human endometrial cancer bioptic specimens the spatial distribution of MCs shows significant deviation from randomness as compared with control group in which, instead, the spatial distribution of MCs is consistent with a random distribution. These findings confirm that MCs enhance tumor angiogenesis and their preferential localization along blood vessels and sites of new vessel formation sustaining the suggestion for an association between MCs and angiogenesis. However, this spatial association between vessels and MCs might simply reflect migrating MCs from the blood stream at vessel growing sites.     

A comparative study of the spatial distribution of mast cells and microvessels in the foetal, adult human thymus and thymoma

Mast cells (MCs) are widely distributed in human and animal tissues and have been shown to play an important role in angiogenesis in normal and pathological conditions. Few data are available about the relationship between MCs and blood vessels in the normal human thymus, and there are virtually no data about their distribution and significance in thymoma. The aim of this study was to analyse the spatial distribution of MCs and microvessels in the normal foetal and adult thymus and thymoma. Twenty biopsy specimens of human thymus, including foetal and adult normal thymus and thymoma were analysed. Double staining with CD34 and mast cell tryptase was used to count both mast cells and microvessels in the same fields. Computer-assisted image analysis was performed to characterize the spatial distribution of MCs and blood vessels in selected specimens. Results demonstrated that MCs were localized exclusively to the medulla. Their number was significantly higher in thymoma specimens as compared with adult and foetal normal specimens respectively. In contrast the microvessel area was unchanged. The analysis of the spatial distribution and relationship between MCs and microvessels revealed that only in the thymoma specimens was there a significant spatial association between MCs and microvessels. Overall, these data suggest that MCs do not contribute significantly to the development of the vascular network in foetal and adult thymus, whereas in thymoma they show a close relationship to blood vessels. This could be an expression of their involvement not only in endothelial cells but also in tumour cell proliferation.     

Spatial distribution of mast cells around vessels and glands in human gastric carcinoma

The spatial distribution of mast cells inside the tumor stroma has been little investigated. In this study, we have evaluated tumor mast cells (MCs) distribution in gastric cancer through the analysis of the morphological features of the spatial patterns generated by these cells, including size, shape, and architecture of the cell pattern. The pattern of distribution of tryptase- and chymase-positive MCs around the blood vessels and gastric glands in human gastric adenocarcinoma samples was investigated by immunohistochemical techniques and by introducing a quantitative approach to characterize the spatial distribution of MCs. In human gastric cancer, both chymase-positive MC and vessels exhibited significant deviations from randomness for what it concerns their spatial relationship with gastric parenchyma. As indicated by cell-to-gland distances shorter than expected by chance, in grade II samples a preferential localization of chymase-positive MC near the gastric glands was observed. Interestingly, the same type of spatial association was exhibited by vessels in grade IV samples, where vessel-to-gland distances shorter than expected by chance were observed. These two findings allow to speculate about a sequence of events in which a subpopulation of MC is first recruited around gastric parenchyma to drive the subsequent development of a vascular support to the tissue.     

Histamine induces the neuronal hypertrophy and increases the mast cell density in gastrointestinal tract

Histamine is an endogenous biogenic amine that is synthesized from the basic amino acid histidine. Ability to mimic anaphylaxis is one of the first described functions of histamine and it has been demonstrated that histamine plays a significant role in the regulation of immune system and neuronal function, influences neuronal morphology and is involved in mast cells (MCs) chemotaxis. MCs as histamine releasers, may thus also interact with neuronal function. In the present study, we aimed to evaluate the role of histamine on mast cell density and neuronal morphology in the gastrointestinal tract of the mouse.Ten mice were daily injected intraperitoneally for 7 days with 20 mg/kg of histamine diluted in 0.5 ml physiological serum. After 7 days, mice were euthanised and samples from stomach, small bowel, colon and appendix were processed for histological examination. Immunohistochemistry was performed employing primary antibodies directed against triptase for mast cells and PGP 9.5 antigen for neuronal structures. The density of triptase and PGP 9.5 positive cells and the morphology of the ganglia were quantitatively evaluated by digital image analysis.The number of ganglia was higher in stomach, small bowel, colon and appendices of the histamine group when compared with the control group. Only in appendices and colon, the number of Schwann cells was significantly higher than that of the control group. The PGP 9.5 expression and the mean area of ganglia showed a significant increase only in appendices. In histamine group the MCs were clustered especially in the lamina propria. Mast cell density (MCD) was significantly higher than the control group in the small bowel, colon and appendices tissues.The intraperitoneally injection histamine increases the MCD and induces the neuronal hypertrophy and after the comparison of the organs in the gastrointestinal tract the results indicated the most effected organ as the appendices.     

Tryptase-positive mast cells correlate with angiogenesis in canine mammary carcinoma

Angiogenesis plays a crucial role in tumour growth, invasion and metastasis. Mast cells (MCs) release angiogenic factors that promote endothelial proliferation and differentiation. Previous studies have suggested that MCs are involved in tumour angiogenesis due to the release of various pro-angiogenic factors. This study evaluated samples from 40 canine mammary carcinomas and eight healthy non-neoplastic canine mammary glands. Toluidine blue staining was performed to characterize the MCs. Immunohistochemical labelling was performed to detect the number of tryptase-positive MCs and microvessels. MCs accumulated in tumour tissue and were closely associated with blood or lymphatic vessels in the tumour microenvironment. Angiogenesis, as measured by microvessel density, increased in direct proportion to the number of MCs. The correlation coefficient was significantly higher for tryptase-positive MCs than for toluidine blue-stained MCs. These results suggest that MCs are involved in tumour angiogenesis, which in turn influences tumour growth, invasion and metastasis. In particular, MC tryptase may be influential in mediating this function of MCs.     

Hyperexpression of FcgammaRI and Toll-like receptor 4 in the intestinal mast cells of Crohn's disease patients

We previously reported that human mast cells (MCs) express high affinity IgG receptor (FcgammaRI) and Toll-like receptor 4 (TLR4) in response to interferon (IFN)-gamma in vitro. The number of MCs is known to increase in Crohn's disease (CD) and ulcerative colitis (UC). We aimed to examine the expression and function of the receptors in these diseases by immunohistochemistry of the colonic mucosae and by in vitro experiments. The density of MCs expressing FcgammaRI, TLR4, or both proteins was significantly higher in CD than in UC or control samples. The density of TNF-alpha(+) MCs expressing FcgammaRI or TLR4 was significantly higher in CD than in control samples. LPS and IgG1 cross-linking synergistically induced a high level of TNF-alpha production in IFN-gamma-treated human MCs. Hyperexpression of FcgammaRI and TLR4 on MCs was related to the high frequency of TNF-alpha expression in CD, suggesting the activation of MCs via these receptors in vivo.     

The frequency and distribution of mast cells in pleomorphic adenomas of salivary glands

AIM: To investigate whether the frequency and distribution of mast cells (MCs) in pleomorphic adenomas (PAs) of major and minor salivary glands justifies the suggestion that there exists an association between MCs and mucoid stromal changes in PAs. METHODS: The material consisted of 22 cases of pleomorphic adenoma (eight arising in major and 14 in minor salivary glands) and a control group represented by five cases of monomorphic adenoma (MA). Representative 3-microm thick, paraffin-embedded sections were stained with H&E and Azur A. Computer-aided image analysis was performed in order to evaluate the relative surface area occupied by epithelial and connective tissue components, as well as the absolute number of MCs. RESULTS: According to our findings, PAs from minor salivary glands contain significantly greater numbers of mast cells compared with tumours from major glands. Additionally, the distribution of MCs within the stromal connective tissue appeared not to be random. CONCLUSION: It is possible that differences in the pattern of connective tissue might influence the actual concentration of MCs and that these differences are responsible for the observed variations between major and minor gland PAs.     

血管内皮生长因子C及其受体3在人恶性黑色素瘤组织内的表达及意义

目的 观察人恶性黑色素瘤组织内血管内皮生长因子C(VEGF-C)及其受体3(VEGFR-3)的表达,探讨VEGF-C和VEGFR-3在恶性黑色素瘤淋巴管生成及淋巴道转移中的作用.方法 取人恶性黑色素瘤组织48例(石蜡标本30例,术后新鲜组织18例),应用免疫组织化学和RT-PCR技术,观察VEGF-C和VEGFR-3蛋白及mRNA在恶性黑色素瘤组织内的表达情况.以淋巴管内皮透明质酸受体(LYVE-1)标记淋巴管,计数恶性黑色素瘤组织淋巴管数密度.结果 VEGF-C和VEGFR-3蛋白主要表达于恶性黑色素瘤细胞胞浆内,在肿瘤周围的血管和淋巴管内皮上也可见VEGFR-3蛋白表达,VEGF-C和VEGFR-3蛋白在淋巴结转移组恶性黑色素瘤组织内的表达水平明显高于无淋巴结转移组(P<0.05).在18例新鲜恶性黑色素瘤中,淋巴结转移组VEGF-C和VEGFR-3mRNA的表达明显高于无淋巴结转移组(P<0.01).LYVE-1表达于肿瘤间质内的淋巴管内皮细胞,淋巴结转移组恶性黑色素瘤组织中的淋巴管数密度(LMVD)为9.845±2.454,无淋巴结转移组恶性黑色素瘤组织中的淋巴管数密度为6.534±2.193,淋巴结转移组恶性黑色素瘤组织内的淋巴管数密度明显高于无淋巴结转移组(P<0.01).结论恶性黑色素瘤组织内VEGF-C表达明显增高,并通过上调其受体VEGFR-3的表达促进恶性黑色素瘤组织内淋巴管的生成,从而促进恶性黑色素瘤的淋巴道转移.     

Number of pericryptal fibroblasts correlates with density of distinct mast cell phenotypes in the crypt lamina propria of human duodenum: implications for the homeostasis of villous architecture

Pericryptal fibroblasts (PFs), a class of myofibroblasts, have strongly been implicated in the regulation of villous structure because of their location close to crypts and their ability to secrete cytokines affecting intestinal epithelial cell proliferation and differentiation. Recently, mast cells (MCs) have also been involved in the homeostasis of villous architecture. As myofibroblasts arise in a wide variety of settings concurrently with a local increase in the number of tissue MCs, we calculated in this study the density of both PF and distinct pericryptal MC phenotypes in the mucosa of human duodenum showing normal, defective, or atrophic villous profiles. In addition, we evaluated the statistical association between PF-MC densities and each pattern of villous architecture. Finally, we correlated the density of PF with the density of pericryptal MC phenotypes. For this purpose, samples taken by endoscopy from 30 patients complaining of inflammatory bowel disorders were studied by immunohistochemistry. The densities of alpha-smooth muscle actin-positive PFs as well as tryptase-, chymase-, and c-kit-positive MCs were determined in the crypt lamina propria. Villous architecture was found to be significantly associated with the number of PFs and tryptase-, chymase-, c-kit-positive MCs in the lamina propria (ANOVA group effect P < 0.001). High density of both PFs and MCs was found in intestinal samples with normal villous morphology while lower densities were associated with defective or atrophic villous profiles (Tukey's test for multiple comparison P < 0.001). In addition, a significant correlation was found between PF density and the density of each pericryptal MC phenotype (vs. tryptase-positive MCs, r = 0.913; vs. chymase-positive MC, r = 0.905; vs. c-kit-positive MC, r = 0.927; P < 0.001 in all cases). This study provides morphological support for an important cooperation between PFs and MCs in maintaining normal villous architecture.     

Increased density of interstitial mast cells in amyloid A renal amyloidosis.

Renal interstitial fibrosis is the final common pathway leading to end-stage renal disease in various nephropathies including renal amyloidosis. However, the role of mast cells (MCs) in the fibrotic process of renal amyloidosis is not fully understood. We compared the distribution of MCs in renal biopsies from 30 patients with AA type renal amyloidosis and 20 control cases. Immunoreactivity of renal MCs to anti-tryptase and anti-chymase was studied. Interstitial myofibroblasts were stained with anti-alpha-smooth muscle actin (alpha-SMA) antibody, and inflammatory cells were identified by anti-CD45, -CD20, and -CD68 mAbs. Positively stained cells were counted, and the relative interstitial and fractional areas of anti-alpha-SMA stained cells were measured. Anti-CD29 mAb was used to detect beta1 integrin and anti-basic fibroblast growth factor (bFGF) mAb for the growth factor on MCs. MCs were rarely found in control samples. In contrast, samples showing amyloid deposition contained numerous tryptase-positive (MCT) (940.17 +/- 5.4 versus 6.74 +/- 1.1/mm2) but fewer chymase-positive (MCTC) cells (20.7 +/- 2.86 versus 1.7 +/- 0.76/mm2) in the renal interstitium. There was a significant relationship between interstitial MCT and creatinine clearance (r = -0.72), and between interstitial MCT and glomerular amyloid-index (GAI) (r = 0.723) and interstitial amyloid area (r = 0.824). Accumulation of MCs correlated significantly with the number of T lymphocytes (MCT: r = 0.694). There was also a significant relationship between mast cell (MC) number and the fractional area of alpha-SMA positive interstitium (r = 0.733) and interstitial fibrotic area (r = 0.6). Double immunostaining demonstrated intracytoplasmic presence of beta1 integrin on 87% of MCT and correlated significantly with the interstitial amyloid area (r = 0.818, P = .001) and T-cell number (r = 0.639, P = .002). bFGF was also detected on 85.5% of MCTC correlating well with the interstitial alpha-SMA-area (r = 0.789). Our results indicate that MCs constitute an integral part of the overall inflammatory process and play a crucial role in interstitial fibrosis in renal amyloidosis.     

小鼠恶性黑色素移植瘤模型的建立及其淋巴管内皮细胞透明质酸受体1的表达

目的:建立小鼠皮肤恶性黑色素移植瘤模型,探讨小鼠皮肤恶性黑色素移植瘤组织内淋巴管的生成情况。方法:体外培养B16瘤细胞,接种B16瘤细胞于C57BL/6小鼠右侧背部皮内,构建C57BL/6小鼠皮肤恶性黑色素移植瘤模型;应用H-E染色、淋巴管内皮细胞透明质酸受体1(1ymphatic vessel endothelial hyaluronan receptor-1, LYVE-1)免疫组化染色确认模型和观察肿瘤组织内淋巴管的生成情况。结果:建立了恶性黑色素移植瘤模型;LY- VE-1主要着色于正常组织和移植瘤组织中淋巴管内皮细胞的细胞膜上;恶性黑色素瘤组织中淋巴管密度明显高于正常皮肤组织。结论:构建C57BL/6小鼠皮肤恶性黑色素移植瘤模型是获得恶性黑色素瘤组织和进一步研究的有效手段;LYVE-1是特异性较高的淋巴管内皮标记物;小鼠皮肤恶性黑色素移植瘤组织中可能存在淋巴管的新生。     

肥大细胞转化生长因子β在人慢性根尖周病组织中的表达

目的:本研究旨在观察不同类型慢性根尖周病组织中肥大细胞(mast cells,MCs)的分布,分析转化生长因子β(transforming growth factor-β,TGF-β)在MCs上的表达情况,探讨类胰蛋白酶(tryptase)-TGF-β双阳性MCs在慢性根尖周病免疫病理中的作用。方法:共收集78例样本,分为3组:(1)根尖肉芽肿组25例;(2)根尖囊肿组25例;(3)正常对照组28例。组织标本经4%甲醛固定48 h以上。石蜡包埋、制备组织连续切片。HE染色,观察其组织学改变;采用甲苯胺蓝染色法观察各组标本组织中MCs的数量及MCs脱颗粒情况;采用免疫荧光双染色法,在荧光显微镜下观察各组标本组织中tryptase-TGF-β双阳性MCs的数量。结果:(1)与正常对照组相比,两组慢性根尖周病组织中tryptase-TGF-β双阳性MCs密度显著增多(P0.01);(2)根尖囊肿组tryptase-TGF-β双阳性MCs密度明显高于根尖肉芽肿组(P0.01);(3)与甲苯胺蓝染色法相比,tryptase-TGF-β免疫荧光双染色法结果中各组标本组织的MCs密度显著增多(P0.01)。结论:慢性根尖周病组织,尤其是根尖囊肿组织中tryptaseTGF-β双阳性细胞明显增多,提示tryptase-TGF-β双阳性MCs可能在慢性根尖周病中的致病过程,尤其在根尖周囊肿纤维组织形成中发挥作用。Tryptase免疫荧光染色法比传统鉴别肥大细胞的甲苯胺蓝染色法更灵敏。     

Impact of different progestins on endometrial vascularisation of postmenopausal women. A retrospective image analysis-morphometric study

OBJECTIVES: To determine if the vascularisation of the endometrium is dependent on the administered progestin during sequential hormone replacement therapy. METHODS: Nine women received percutaneous estradiol-17 beta, 1.5 mg/day from days 1 to 24 combined with 200 mg/day micronised progesterone from days 11 to 24 of the treatment cycle. Fifteen women received percutaneous estradiol, 1.5 mg/day from days 1 to 24, combined with 10 mg/day chlormadinone acetate from days 11 to 24. Eleven women received percutaneous estradiol, 50 microg/day from days 1 to 28 combined with percutaneous norethisterone acetate, 0.3mg/day form days 14 to 28. Twelve women received intranasal estradiol, 300 microg/day from days 1 to 25 combined with 0.5 mg of promegestone from days 11 to 24. Eleven spontaneous cycling women had an endometrial biopsy during luteal phase and served as controls. Endometrial biopsies were processed routinely between days 18 and 24 and sections were immunostained using anti-CD34 antibody to identify vascular endothelial cells, which were treated with an automatic image analysis system. RESULTS: mean (+/-S.D.) vascular density for controls was 147+/-41.5 vessels/mm(2), with mean vessel area of 143+/-60.9 microm(2). In chlormadinone users endometrial microvascular density and mean vessel area did not differ from the control group (150.2+/-58.6 and 152.9+/-70.5). The other three progestins generated a significant increase of mean vessel density, 179.6+/-51.6 with micronised progesterone, 178.5+/-67.6 with norethisterone and 179.6+/-48.4 with promegestone. The mean vessel area was lower in the latter three groups, respectively, 108.4+/-39.0, 97.5+/-46.5 and 141.6+/-66.7 microm(2), promegestone leading to non significant difference with control. CONCLUSION: regarding vascularisation, chlormadinone and control group gave similar patterns. Promegestone was associated with an increase of the number of vessels, as did micronised progesterone or norethisterone; the mean vascular area was the smallest in the norethisterone group.     

Tissue hemostasis and chronic inflammation in colon biopsies of patients with inflammatory bowel disease

Inflammatory bowel disease (IBD) is characterized by a chronic inflammation accompanied by procoagulation settings. However, tissue hemostasis in IBD patients was only incidentally reported. Accordingly, the current study characterizes changes in tissue hemostasis components in a colon inflammatory setting. Serial cryostat sections of endoscopic mucosal biopsy specimens taken from 26 consecutive IBD patients diagnosed de novo and normal colon resection specimens taken from 6 patients were immunohistochemically stained with monoclonal anti-human tissue factor (TF), tissue factor pathway inhibitor (TFPI), thrombomodulin (TM), as well as CD3 and CD68 positive cells. The hemostatic components studied differed significantly from the control subjects. Up-regulation predominated in the case of TF while down-regulation was mainly found in TM and TFPI in IBD. In the control sections, TF was observed in a few fibroblast-shaped cells in the lamina propria, while in the majority of IBD sections, TF positively stained small microvessels, infiltrating mononuclear cells and fibroblast-shaped cells tightly surrounding the colon crypts. Thrombomodulin intensively stained the endothelium of the small capillary vessels in the control, whereas such staining mainly accompanied infiltrating mononuclear cells of the IBD subjects. Tissue factor pathway inhibitor positively stained the endothelium of the small capillary vessels in the control group, whereas in the IBD group endothelial cells presented only weak TFPI staining. The mean number of CD3-positive lymphocytes in IBD was 23.3±14.3, but the mean number of CD68-positive cells was 114.5±55.8. In the control sections, it was 4.1±2.4 and 39.6±17.9, respectively. There was no relationship between CD3 and CD68 (+) cells and the hemostasis markers studied. The results of the current study indicate a shift of tissue hemostasis toward the procoagulant state irrespective of the severity of inflammatory infiltration. In addition, TF distribution in the colon sections of IBD patients may indicate a role in the restoration of the barrier function in injured intestinal mucosa.     

Identification and characterization of survivin‐derived H‐2Kb‐restricted CTL epitopes

Survivin is overexpressed in several malignancies and in tumor‐associated endothelium making it an attractive target for therapeutic cytotoxic T‐cell responses. Thus, it would be important to test this notion in preclinical models. Consequently, we screened the murine survivin sequence for potential binding K^b‐restricted octamer peptide epitopes. Two epitopes, which bind strongly to K^b, were selected to test their immunogenicity in vivo. Spleen cells from mice vaccinated by intradermal injection of mature DC pulsed with these peptides displayed reactivity to the respective epitopes. The natural processing and presentation of these epitopes by tumor cells was evident by the killing of murine melanoma cells by vaccination‐induced T cells. Subcutaneous challenge with syngeneic melanoma demonstrated the protective immunity of this vaccination. Notably, analysis of the vessel density in subcutaneous tumors revealed that survivin‐specific vaccination significantly reduced the number of intratumoral vessels. In summary, we demonstrated the immunogenicity of two K^b‐restricted peptide epitopes derived from the murine survivin protein; moreover, survivin‐specific vaccination not only resulted in a reduction of tumor cells but also the tumor supplying blood vessels. The presented preclinical model for survivin‐directed vaccination may serve as a valuable tool to improve already running clinical trials in a syngeneic tumor model.     

Morphological features of regeneration of rabbit aortic endothelium after cryoinduced vascular damage

Regeneration of endothelium after damage is an important factor, which limits the development of atherogenesis. This study examines the topographical characteristics of regenerating endothelial cells (EC) in rabbit aorta after de-endothelialization by cryodestruction. The effects of cloricromene on these processes were also studied. Vessels were harvested from 6-month-old NZW rabbits, 1 and 3 days after cryodestruction. The vessels were evaluated using scanning electron microscopy (SEM). One day after cryodestruction, there were defects in the endothelial monolayer in the zone of injury in saline treated animals. Large numbers of platelets and monocytes were observed in association with endothelium in the damaged zone. Three days after de-endothelialization the size of the area of the damage had decreased. On the surface of the new endothelial layer and below this defect, the number of adhering monocytes was increased, and many microdefects between endothelium could be seen. Administration of cloricromene for 1 or 3 days after damage reduced the number of endothelium-adherent platelets and monocytes, and microdefects in endothelium. The feature of endothelial repair in rabbits is a relatively large involvement of monocytes and platelets, which are visible below regenerated endothelium. Administration of cloricromene essentially restored re-endothelialization and significantly decreased the number of adherent monocytes and microdefects in the new endothelium.     

Clinical significance of the histomorphometric evaluation of diabetic microangiopathy in the oral mucosa

Several structural parameters of the capillary vessels were measured in the oral mucosa of patients with diabetes mellitus of type 1 (D.1) and of type 2 (D.2), and of control cases (C), by means of an image analyser in histological sections of routinely processed biopsies. The studied parameters included: a) capillary wall thickness; b) capillary diameter; c) the ratio of capillary wall thickness and diameter; d) capillary wall area; e) capillary area; f) the ratio of capillary wall area and capillary area; g) density of capillary vessels in the lamina propria; h) density of endothelial cells; i) endothelial nuclear area. Clinical and laboratory parameters were also evaluated (duration of the disease, systolic and diastolic blood pressure levels, glycemia, glycosylated haemoglobin, glycosylated albumin, fructosamines, apolipoproteins A1 and B), in order to assess whether a relationship exists with the morphometric parameters studied. Statistically significant differences, at the level of p less than 0.05, were found in the following morphometric parameters between controls and each group of diabetic patients: mean and standard deviation of capillary wall thickness, mean capillary wall area, mean ratio of the capillary wall area and capillary area. A reduction in the capillary density, i.e. the number of capillary vessels per mm2 of lamina propria, was also observed in diabetic patients with respect to the control group, although it was not statistically significant (C vs. D.1: p less than 0.21; C vs. D.2: p less than 0.10).(ABSTRACT TRUNCATED AT 250 WORDS)     

Tumor lymphangiogenesis: a novel prognostic indicator for cutaneous melanoma metastasis and survival

Malignant melanomas of the skin are distinguished by their propensity for early metastatic spread via lymphatic vessels to regional lymph nodes, and lymph node metastasis is a major determinant for the staging and clinical management of melanoma. However, the importance of tumor-induced lymphangiogenesis for lymphatic melanoma spread has remained unclear. We investigated whether tumor lymphangiogenesis occurs in human malignant melanomas of the skin and whether the extent of tumor lymphangiogenesis may be related to the risk for lymph node metastasis and to patient survival, using double immunostains for the novel lymphatic endothelial marker LYVE-1 and for the panvascular marker CD31. Tumor samples were obtained from clinically and histologically closely matched cases of primary melanomas with early lymph node metastasis (n = 18) and from nonmetastatic melanomas (n = 19). Hot spots of proliferating intratumoral and peritumoral lymphatic vessels were detected in a large number of melanomas. The incidence of intratumoral lymphatics was significantly higher in metastatic melanomas and correlated with poor disease-free survival. Metastatic melanomas had significantly more and larger tumor-associated lymphatic vessels, and a relative lymphatic vessel area of >1.5% was significantly associated with poor disease-free and overall survival. In contrast, no differences in the density of tumor-associated blood vessels were found. Vascular endothelial growth factor and vascular endothelial growth factor-C expression was equally detected in a minority of cases in both groups. Our results reveal tumor lymphangiogenesis as a novel prognostic indicator for the risk of lymph node metastasis in cutaneous melanoma.     

皮肤肿瘤组织中类高内皮微静脉的形态学研究

目的：探讨人黑色素瘤组织中类高内皮微静脉与肿瘤浸润淋巴细胞（tumor infiltrating lymphocyte，TIL）的形态特征。方法：采用光镜和透射电镜观察了lO例手术切除的人皮肤黑色素瘤组织中的类高内皮微静脉和TIL的分布和超微结构。结果：类高内皮微静脉主要分布于乳头层和癌周围。病变早期其内皮高大，胞浆突起增多，核大，细胞器丰富，有大量淋巴细胞穿越管壁；病变晚期，内皮细胞核不规则，出现大量齿状切迹，细胞质内出现大量微丝和质膜小泡，线粒体多出现肿胀、扩张，基膜不连续，少见淋巴细胞穿越。TIL多存在于癌周围区，活化淋巴细胞较多。随着病程的进展，淋巴细胞数世减少：结论：1．皮肤黑色素瘤组织中部分血管可演变为类高内皮微静脉，是淋巴细胞向癌组织浸润（淋巴细胞定向归巢）的重要场所：2．肿瘤组织中的淋巴细胞归巢与抗肿瘤密切相关。     

白细胞介素1β和白细胞介素17在根尖周病组织肥大细胞上的表达

 目的：观察不同类型慢性根尖周病组织中肥大细胞（mast cells, MCs）的分布情况，并分析白细胞介素1β（interleukin-1β，IL-1β）和白细胞介素17（interleukin-17，IL-17）在MCs上的表达情况，探讨MCs IL-1β和IL-17在根尖周病发病机制中的作用。方法：本实验共收集102例样本，分为3组：（1）根尖肉芽肿组32例；（2）根尖囊肿组35例；（3）正常对照组35例。组织标本经4%中性甲醛缓冲液固定48 h以上；石蜡包埋，组织连续切片，HE染色，在光学显微镜下观察其组织学改变；采用免疫荧光双染色法，在荧光显微镜下观察各组标本组织表达IL-1β和IL-17的MCs数目。结果：两组慢性根尖周病的炎症反应程度明显高于正常对照组（P<0.01）；根尖囊肿组与根尖肉芽肿组之间炎症反应程度无显著差异（P>0.05）。与正常对照组相比，两组慢性根尖周病组织中IL-1β和IL-17阳性MCs数目均显著增多（P<0.01）；根尖囊肿组与根尖肉芽肿组IL-1β和IL-17阳性MCs密度无显著性差异（P>0.05）。经Pearson相关分析，3组标本组织中IL-1β和IL-17阳性MCs密度与组织炎症反应程度存在直线的正相关关系（P<0.01）。结论：慢性根尖周病组织中MCs数目明显增多，同时IL-1β和IL-17阳性的MCs密度显著增高。IL-1β和IL-17阳性MCs密度与根尖周病的炎症程度趋势相一致。这提示IL-1β和IL-17阳性MCs可能在慢性根尖周病的致病机制中发挥重要作用。