EXPRESSION OF ICAM-1 AND VCAM-1 IN HUMAN MALIGNANT MESOTHELIOMA

Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are cytokine-inducible adhesion molecules which recognize ligands that are highly expressed on leukocytes. Expression of ICAM-1 and VCAM-1 was investigated in tissue sections of 16 cases of malignant mesothelioma (seven epithelial, eight biphasic, and one sarcomatoid) using immunohistochemistry. Neoplastic cells were diffusely and intensely stained for ICAM-1 in all cases. VCAM-1 was detected in 14 of 16 cases. The percentage of VCAM-1-positive tumour cells was more than 50 per cent in eight cases and the staining was observed mainly in epithelial-like cells. VCAM-1 was rarely expressed in other malignant tumours of epithelial origin, being present in only 1 of 58 cases of carcinoma originating from different anatomical sites. At the cellular level, ICAM-1 and VCAM-1 appeared co-distributed, the staining for both being cytoplasmic with a membrane reinforcement. The regulation of VCAM-1 expression by neoplastic mesothelial cells was investigated in vitro using 14 mesothelioma cell lines. ICAM-1 was expressed by cultured cells of all mesothelioma cell lines, even in the absence of cytokines. VCAM-1 was detected in 10–50 per cent of the cells in three non-stimulated mesothelioma cell lines (mero-95, mero-96, and mero-134), and was absent or poorly expressed in the remaining 11. Exposure of a negative cell line (mero-48a) to an optimal concentration of tumour necrosis factor alpha (TNFα) or interleukin-13 (IL-13) for 6–18 h resulted in the induction of VCAM-1 mRNA synthesis and in VCAM-1 expression at the membrane level in 60–70 per cent of the cells. These findings are consistent with the possibility that TNFα, IL-13, or other activating signals are released in the tumour micro-environment and regulate the expression of VCAM-1 in malignant mesothelioma cells.     

IMMUNOHISTOCHEMICAL STUDY OF OVEREXPRESSION OF FIBROBLAST GROWTH FACTOR-1 (FGF-1), FGF-2, AND FGF RECEPTOR-1 IN HUMAN MALIGNANT SALIVARY GLAND TUMOURS

Fibroblast growth factor-1 (FGF-1) and FGF-2 are broad spectrum mitogens. The expression of FGF-1, FGF-2, and their receptor, FGF receptor-1 (FGFR-1), was examined in malignant salivary gland tumours and normal salivary glands, using immunohistochemical methods. In seven cases of adenoid cystic carcinoma (ACC), both duct-like cells and modified myoepithelial cells were apparently immunopositive for FGF-1, FGF-2, and FGFR-1. In five cases of mucoepidermoid carcinoma (MC), all three types of tumour cells including epidermoid cells, mucous cells, and intermediate cells expressed immunoreactive FGF-1, FGF-2, and FGFR-1. In these malignant salivary gland tumours, increased expression of FGFR-1 correlated with the intensity of both FGF-1 and FGF-2 immunoreactivity. In contrast to malignant salivary gland tumours, eight cases of normal salivary gland showed negative immunostaining for FGF-1, FGF-2, and FGFR-1 while four cases were weakly immunoreactive for FGF and its receptor. These results demonstrate that malignant salivary gland tumours overexpress FGF-1, FGF-2, and FGFR-1 compared with normal salivary glands and suggest that these growth factors may play an important role in facilitating neoplastic progression in human salivary glands.     

EVALUATION OF TUMOUR ANGIOGENESIS AS A PROGNOSTIC MARKER IN MALIGNANT MESOTHELIOMA

Angiogenesis plays an important role in the growth, progression, and metastasis of solid tumours. Malignant mesothelioma (MM) of the pleura is a highly invasive tumour with a poor prognosis. In the present study, microvascular quantification was undertaken on 25 specimens of mesothelioma and 15 specimens of non-neoplastic mesothelium (NNM), by staining for the antigens CD34 and CD31. Areas of highest intratumoural microvascular density (IMD) were identified and counted either manually (mIMD) or on a computerized image analysis system (CIAS; iIMD). The two IMDs were significantly correlated with each other ( r =0·736; P <0·001). The average IMD for MM was significantly ( P <0·001) higher than in NNM. Moreover, each unit increment in iIMD for MM, when regarded as a continuous variable, was significantly ( P =0·001) associated with an increased hazard of about 4 per cent. When regarded as a categorical variable, the patients in the highest tertile (>58 vessels/field) had a significantly ( P <0·01; log-rank test) shorter survival than patients in the lowest tertile (<45 vessels/field). This association was independent of the age of the patient and of the histological type or grade of the MM. No association was noted with p53 immunoexpression. Although the mean vascular area of blood vessels measured on the CIAS did not correlate with survival, assessment of IMDs can be an important independent prognostic indicator in malignant mesothelioma. © 1997 John Wiley & Sons, Ltd.     

运动神经诱向因子1及其受体在大鼠下颌下腺的表达

实验用运动神经诱向因子１的单克隆抗体及抗独特型单克隆抗体的免疫组织化学染色研究了运动神经诱向因子１及其受体在大鼠下颌下腺的表达。邻片免疫组织化学法显示：成年雄性大鼠下颌下腺的浆液性腺泡及各级导管上皮细胞增多呈运动神经诱向因子１及其受体的免疫的应阳性，副交感神经节细胞也呈运动神经诱向因子１及其受体免疫反应阳性，提示下颌下腺的以上结构均能自分泌运动神经诱向因子１。生后１日龄大鼠、仅见个别腺导管显示为极     

人骨髓巨核细胞分化发育过程中神经生长因子基因表达

实验通过原位分子杂交技术,检测人骨髓巨核细胞分化发育过程中神经生长因子（ＮＧＦ）基因表达情况。结果显示β神经生长因子ＤＮＡ探针与其互补的ｍＲＮＡ杂交后应信号存在于各级不同发育的巨核细胞胞浆中。原巨核数量很少,较难发现,杂交后反应阳性的胞浆很少,呈线状。幼巨核细胞阳性胸浆较少,杂交反应产物分布较为均匀。颗粒型巨核细胞胞浆丰富,杂交反应产物深浅不一,常见较为细小的阳性颗粒,成就型巨核细胞体积很大,胞浆     

EXPRESSION OF THE TYPE 1 TYROSINE KINASE GROWTH FACTOR RECEPTORS EGF RECEPTOR,c-erbB2 AND c-erbB3 IN BLADDER CANCER

Expression of the c-erbB3 protein was determined in transitional cell carcinoma of the bladder by immunohistochemistry. Strong membrane staining was observed in 10 per cent of cases (7/70) and cytoplasmic and membrane overexpression in 20 per cent (14/70). Overexpression of the epidermal growth factor (EGF) receptor (36 per cent, 25/70) and c-erbB2 proteins (9 per cent 6/70) was determined in the same series of cases. c-erbB3 overexpression was positively correlated with EGF receptor expression ( P <0·025) but appeared to be inversely associated with c-erbB2 overexpression.     

人血管内皮生长因子基因的克隆、表达及生物活性分析

目的 克隆人血管内皮细胞生长因子165(VEGF165)基因,构建真核表达载体,观察其对脐静脉内皮细胞的增殖作用和血管新生的影响。方法 利用RT-PCR方法,从人扁桃体组织中扩增人VEGF165 cDNA完整编码区,并构建成pcDNA3．1( )／VEGF165(简称pcDNA／V)重组体;应用脂质体介导的基因转移技术将构建的真核表达载体pcDNA／V体外转染至人脐静脉内皮细胞(HUVEC),MTT法检测其对内皮细胞增殖的影响。建立家兔下肢缺血模型,注射重组质粒pcDNA／V,pcDNA3．1( )空质粒作对照,选取不同时间点,行血管造影。结果 构建的真核表达载体pcDNA／V的酶切电泳分析和测序表明结果正确。pcDNA／V转染HUVEC能明显促进内皮细胞的分裂增殖。血管造影显示,术后基因治疗组远端动脉充盈早于对照组,新生血管数目也明显多于同时期对照组。结论 成功克隆了人VEGF165基因,构建了其真核表达载体。体内外生物学活性研究证实,重组质粒的表达产物具有刺激HUVEC增殖和促进缺血肢体侧枝循环建立的功能。     

IMMUNOREACTIVITY FOR HEPATOCYTE GROWTH FACTOR/SCATTER FACTOR AND ITS RECEPTOR,met, IN HUMAN LUNG CARCINOMAS AND MALIGNANT MESOTHELIOMAS

Paraffin sections from 29 lung carcinomas (28 primary and 1 metastatic) and 9 pleural malignant mesotheliomas were immunostained with antisera to human hepatocyte growth factor/scatter factor (HGF/SF) and its receptor, met. For HGF/SF, immunoreactivity was demonstrated in all 9 mesotheliomas, 9 of 12 adenocarcinomas, and 7 of 10 squamous cell carcinomas. None of seven cases of small cell anaplastic carcinoma was positive. The adenocarcinomas frequently showed enhanced luminal staining, suggesting possible secretion of HGF/SF, and this pattern of staining was also seen occasionally in bronchial epithelium adjacent to the tumour. Stromal fibroblasts also showed immunoreactivity for HGF/SF in 6/8 cases of mesothelioma but in only 3/12 adenocarcinomas, 1/10 squamous cell carcinomas, and 1/4 small cell anaplastic carcinomas. All tumours stained for met, usually strongly. The staining was mainly cytoplasmic in nature, but some plasma membrane staining was usually evident. Adenocarcinomas showed strong luminal membrane staining, as did adjacent, histologically normal bronchial epithelium. This study demonstrates the presence of HGF/SF and met in most of the tumour types described, particularly mesotheliomas, and suggests that the HGF/SF/met signalling system may play a role in the development of these tumours, either by autocrine or by paracrine mechanisms.     

脑缺血再灌注后脑梗塞周围区生长相关蛋白-GAP43表达的变化及外源性神经生长因子的影响

目的研究大鼠脑缺血再灌注后生长相关蛋白-43(GAP-43)表达的变化规律及外源性神经生长因子(NGF)的影响.方法成年健康雌性Wistar大鼠72只,随机分为假手术组、自然恢复组、人工脑脊液组和NGF治疗组.采用线栓法建立大脑中动脉缺血再灌注(MCAO)动物模型,应用免疫组织化学方法观察脑缺血再灌注后GAP-43的表达.结果(1)脑缺血再灌注6 h后,缺血周围区GAP-43表达逐渐增高,第7天达高峰,以后逐渐降低,第21天仍有表达.(2)应用外源性NGF后,GAP-43表达较对照组有所增高,但无显著性差异(P＞0.05).结论提示中枢神经系统损伤后,神经元具有再生和修复的可塑性,外源性NGF对GAP-43的调节有待于进-步研究.     

血管内皮生长因子及其受体在糖尿病小鼠视网膜的表达

目的：观察血管内皮生长因子（Vascular Endothelial Growth Factor,VEGF)及其受体flk-1在糖尿病视网膜细胞的表达,以期阐明VEGF与糖尿病性盲的关系。方法：应用免疫组织化学这及发病组视网膜VEGF和flk-1阳性细胞,并计算单位面积视网膜中神经节细胞VEGF阳性率和flk-1阳性细胞数,结果：发病组视网膜神经节细胞VEGF阳性率和flk-1阳性神经节细胞数从3月龄时开始增多（P＜0．05）,并随着病程延长有上升趋势,VEGF在6月龄后增多更显著,flk-1在8月龄后增加也更为明显。结论：VEGF和flk-1在糖尿病视网膜神经节细胞表达量增加,VEGF通过与flk-1相互作用,不仅使血管内皮细胞增殖,导致视网膜新生网管形成,而且还可能通过改变神经节细胞及Muller细胞膜上的受体数量而对神经节细胞功能活动产生影响。     

大鼠肝癌发生过程中转化生长因子—α在肝组织中的表达

以二乙基亚硝胺诱发大鼠肝癌、用免疫组织化学法检测肝组织中转化生长因子－α的表达变化。结果显示：正常大鼠肝的枯否细胞呈转化生长因子－α阳性表达；至诱癌第８周，除少量枯否细胞仍呈转化生长因子－α阳性外，部分肝细胞亦呈转化生长因子－α阳性，且吾两种反应类型，即部分肝细胞质和胞膜均呈转化生长因子－α阳性。有的肝细胞仅胞膜呈转化生长－α阳性。随着肝癌病变发展，转化生长因子－α阳必肝细胞数渐增多，转化生长因子     

人胎海马生长抑素和神经生长因子免疫阳性神经元的研究

本文采用免疫细胞化学ABC法对20例胎龄为16～36周的人胎儿海马进行了观察。证明人胎海马存在着生长抑素和神经生长因子免疫阳性神经元，两者均散在分布于海马皮质的深层，为多极神经元；免疫反应阳性颗粒均匀地分布于神经元胞质中，生长抑素阳性细胞中的颗粒比神经生长因子阳性细胞中的颗粒粗大。本文结果提示：人胎海马中存在的上述两类阳性神经元可能在中枢神经系统的发育中起着重要作用。     

转化生长因子β1在大鼠肝内的原位表达

为探讨转化生长因子β１（ＴＧＦβ１）对肝细胞增殖的调节作用，用ＴＧＦβ１单克隆抗体和免疫组织化学方法研究ＴＧＦβ１在大鼠胎肝、正常成年肝、再生肝和癌变肝中的表达。结果表明：正常成年大鼠肝的大部分血管内皮细胞及胆管上皮细胞表达ＴＧＦβ１。大鼠胎肝的少量血窦内皮细胞表达ＴＧＦβ１，出现后表达逐渐增强，至生后３０ｄ的表达水平同成年肝。肝大部分切除１￣２ｄ，血窦内皮细胞ＴＧＦβ１表达增强，于大部切除后１０     

神经生长因子在大鼠心内神经节的表达

目的 观察成年和中老年大鼠心内神经节神经生长因子(NGF)阳性神经元的存在及其变化。方法 免疫组织化学法。结果 成年及巾老年大鼠心内神经节均存在NGF阳性神经元,但中老年大鼠心内神经节NGF阳性神经元明显减少,表达明显降低。结论 心房后壁心内神经节存在NGF阳性神经元,并随年龄增加表达减少,提示NGF表达的减少可能与心脏功能的退化相关。     

寻常性银屑病患者皮损及尿液中表皮生长因子的检测

应用＾１２５Ｉ人类表皮生长因子放射免疫分析法对３０例寻常性银屑病患者及２０例正常人皮损及尿液中ＥＧＦ进行了检测。结果显示：银屑病进行期皮损表皮中ＥＧＦ含量显著高于正常人及恢复期表皮和未受累表皮。而银屑病患者尿液中ＥＧＦ在银屑病各期及正常人之间比较均无显著性差异。提示ＥＧＦ可能在银屑病表皮的过度增殖及异常分化中起重要作用。     

雄激素对大鼠心内神经节神经生长因子表达的影响

目的 观察神经生长因子(NGF)在成年雄性大鼠、雌性大鼠、雌性大鼠睾酮处理组、睾丸切除大鼠、睾丸切除后睾酮替代大鼠心房后壁心内神经节的表达及变化。方法 免疫组织化学染色。结果 各组大鼠心内神经节均存在NGF阳性神经元,但睾丸切除组大鼠心内神经节NGF阳性神经元的数量明显减少,表达明显降低。结论 心内神经节细胞内含NGF;雄激素可能影响心内神经节细胞的NGF表达。     

表皮生长因子受体与雌,孕激素受体在乳腺癌组织中的表达及相互关系

本文应用碘标记的表皮生长因子，用分步离心提取部细胞膜蛋白，以标准的ＥＧＦ作竞争剂，采取放射标记结合法测定了４０例乳腺癌组织中的表皮生长因子受体水平，同时采用葡聚糖活性碳分析法测定了乳腺癌组织中的雌激素受体，孕激素受体以探讨ＥＧＦＲ表达与ＥＲ，ＰＲ在乳腺癌组织中的生物学行为及相互关系。     

血管内皮生长因子C在乳腺癌中的表达及其与淋巴结转移的关系

目的：检测血管内皮生长因子C（VEGF－C）在乳腺癌中有无表达及其表达程度与癌细胞淋巴结转移的关系,以探讨VEGF－C在肿瘤淋巴道转移中的作用以及肿瘤细胞通过淋巴道转移的机理。方法：选用67例行乳腺癌根治术或改良根治术后女患者肿瘤组织石蜡标本,采用免疫组化方法检测其VEGF－C蛋白的表达,结果：在部分乳腺癌的癌细胞胞浆中可检测到VEGF－C蛋白的表达（28／67）,且VEGF－C在伴腋淋巴结转移的病例组中的表达明显高于无腋淋巴结转移的病例组,结论：VEGF－C表达于乳腺癌中,且其表达与乳腺癌肿瘤细胞的淋巴结转移密切相关。     

人胰岛素样生长因子-1基因的克隆、表达和活性检测

目的 克隆人胰岛素样生长因子Ⅰ型(hIGF-1)，构建真核表达载体并进行表达及活性检测，为IGF-1基因治疗糖尿病奠定基础。方法 提取胎儿肝脏总RNA，RT-PCR法扩增IGF-1cDNA片段，重组于pUCM-T载体，测序正确后构建表达载体，转染猴肾成纤维细胞系COS-7，用原位杂交和免疫组织化学检测表达，收集培养上清以胰岛素刺激释放实验进行活性测定。结果 扩增得到710bp带有Kozak序列的IGF-1cDNA片段；成功构建了真核表达载体pCI-neo／hIGF-1；IGF-1在COS-7细胞得到了表达，并具有刺激胰岛素分泌的活性。结论 新构建的载体pCI-neo／hIGF-1能在COS-7细胞中表达、分泌，且所分泌的IGF-1具有生物活性。