肝细胞癌组织微血管密度与血小板源性生长因子BmRNA表达的关系

目的初步探讨肝细胞癌组织中血小饭源性生长因子B（PDGPB）mRNA的表达与微血管密度（MVD）之间的关系。方法用RT-PCR法柃测51例肝细胞癌和20例正常肝组织PDGF-BmRNA。采用免疫组化（sABC）法检测MVD的表达。结果肝细胞癌组织PDGFBmRNA的表达量明显高于正常肝组织，两者比较有显著性差异（P〈0．01）。肝癌组织和正常肝组织的MVD值分别是29．43±4．818和14．56±1．96（P〈0．01）。结论PDGF-B作为一个血管内皮细胞生长调节因子，在肝癌的发生、发展过程中可能起着重要的促进生长和转移作用。     

血管内皮生长因子与结肠癌的相关性研究

应用血管内皮生长因子（VEGF）、血管活性肠肽（VIP）抗体,采用Verhoff铁苏木素染色法、免疫组化SABC染色法对90例手术切除的结肠癌患者和10例结肠正常组织进行血管标记和染色。结果结肠癌微血管密度（MVD）、VEGF表达强度与正常对照组比较有显著性差异（P〈0．01）,且MVD与VEGF表达呈正相关（r=0．590）,VIP在结肠癌组织中较正常结肠组织有较强的表达（P〈0．01）。提示VIP可作为结肠癌的检测指标之一。     

食管鳞癌组织中NF-κB p65、VEGF的表达及其与血管密度测定的意义

采用免疫组化S-P法检测61例食管鳞癌患者组织中核转录因子-κB（NF-κB）p65和血管内皮生长因子（VEGF）的表达及CD105标记的肿瘤内徽血管密度（MVD）。结果NF-κB p65表达与食管鳞癌的组织学分级、浸润深度和淋巴结转移均呈正相关（P均〈0．01）,VEGF蛋白表达及MVD与食管鳞癌组织学分级和浸润深度均无关,但与淋巴结转移呈正相关（P〈0．05）;NF-κB p65与VEGF表达呈正相关（P〈0．01）;二者均与MVD呈正相关（P均〈0．05）。提示NF-κB p65参与食管鳞癌的发生、发展,其机制可能与VEGF的表达和肿瘤血管生成有关。     

慢性肝炎和原发性肝癌患者血管生成素的表达比较及其与血管生成之间的关系

目的探讨原发性肝癌和慢性肝炎组织中血管生成素-1（Ang-1）、血管生成素-2（Ang-2）的表达与血管生成间的关系。方法应用免疫组织化学方法对26例原发性肝癌组织以及58例慢性肝炎组织中Ang-1、Ang-2及CD34标记的微血管密度（MVD）进行检测。结果原发性肝癌组织中Ang-2表达明显高于慢性肝炎组织,差异有显著性（P〈0．05）;而Ang-1在两组间比较无显著性差异（P〉0．05）。原发性肝癌组织中MVD水平明显高于慢性肝炎组织,两组比较差异有显著性（P〈0．05）。相关性分析表明,Ang-2与原发性肝癌肿瘤大小、分级、有无转移、MVD水平相关。慢性肝炎组织中Ang-1、Ang-2表达随着炎症程度分级和纤维化程度分期越高表达越强,但将各分期合并为不同的组合后,各组间比较无显著性差异（P〉0．05）。结论血管生成在慢性肝炎发展到肝癌中起重要作用,但Ang-1、Ang-2作为一种重要的血管生成因子,其在促进慢性肝炎发展到肝癌中的证据还不是很充分,机制还有待进一步研究。     

星形细胞肿瘤组织中VEGF—A、VEGF—C的表达变化及其与肿瘤血管生成的关系

目的观察血管内皮生长因子（VEGF）．A和VEGF—C在星形细胞肿瘤组织中的表达变化,分析两指标间及与肿瘤血管生成的关系。方法留取93例星形细胞肿瘤患者肿瘤组织及9例接受高血压脑出血开颅手术患者的正常脑组织,用免疫组织化学（sP）方法检测VEGF-A、VEGF-C蛋白表达和微血管密度（MVD）,RT-PCR法检测VEGF．A、VEGF．CmRNA表达,对两VEGF间及与肿瘤WHO分级、MVD的关系进行统计学分析。结果星形细胞肿瘤组织中VEGF．A、VEGF．C蛋白阳性表达率及MVD均显著高于正常脑组织,且均随肿瘤恶性程度增高而增强（P均〈0．叭）;星形细胞肿瘤组织中VEGF—A、VEGF-C蛋白表达及两者与MVD均呈显著相关（P均〈0．01）。星形细胞肿瘤组织中VEGF—A和VEGF．CmRNA表达均显著高于正常脑组织,且均随肿瘤恶性程度增高而增强（P均〈0．05）;肿瘤组织中VEGF．A和VEGF．CmRNA表达呈显著相关（P均〈0．05）。结论VEGF—A、VEGF—C在星形细胞肿瘤组织中的表达上调并与肿瘤恶性程度有关,两者间及与肿瘤血管生成均关系密切。     

肺腺癌组织中VEGF-D的表达变化及意义

目的观察肺腺癌组织中血管内皮生长因子D（VEGF—D）的表达,探讨其意义。方法分别采用RT-PCR法、免疫组化法检测48例肺腺癌组织中的VEGF—D mRNA和VEGF—D、微淋巴管密度（MLVD）、微血管密度（MVD）。结果VEGF—D mRNA在肺腺癌组织中的表达高于正常肺组织（P〈0．01）,VEGF—D蛋白阳性率肿瘤周边显著高于肿瘤中心（P〈0．01）;其表达与肿瘤分化、MVD无关,与TNM分期、MLVD、淋巴结转移有关（P均〈0．05）。结论肺腺癌组织中VEGF—D高表达,与淋巴管的生成及转移有关。     

血管内皮生长因子和血管生成与大肠癌发展的关系

目的：探讨血管内皮细胞生长因子（VEGF）和血管生成与大肠癌发展的关系。方法：应用免疫组化法,检测102例大肠癌组织VEGF蛋白表达和微血管密度（MVD）,分析VEGF和MVD及其与大肠癌组织学分级、浸润深度、Dukes分期、淋巴结转移、肝转移和预后的关系。结果：VEGF阳性者MVD值显著高于阴性者（P＜0．01）,VEGF表达和MVD与大肠癌Dukds分期、淋巴结转移和肝转移密切相关（P均＜0．01）,VEGF表达阳性或高MVD的大肠癌患者5年生存率较低（P＜0．01）。结论：VEGF与大肠癌的血管生成密切相关,对大肠癌的生长和浸润转移有促进作用,VEGF和MVD可作为反映大肠癌生物学行为的客观指标。     

VEGF和Ang-2与甲状腺乳头状癌血管生成和转移的相关性探讨

采用免疫组化S-P法检测20例甲状腺乳头状癌（PTC,A组）、15例结节性甲状腺肿（B组）、15例甲状腺腺瘤（C组）和15例正常甲状腺组织（D组）中血管内皮生长因子（VEGF）、促血管生成素-2（Ang-2）蛋白的表达和微血管密度（MVD）。结果A组VEGF和Ang-2表达均显著高于其他组（P均〈0．05）;A组VEGF、Ang-2与MVD呈正相关（P均〈0．05）,而Ang-2与MVD无相关性;A组有淋巴结转移者VEGF表达和MVD均显著高于无转移者（P均〈0．05）,但Ang-2无显著差异。提示VEGF和Ang-2在PTC血管生成中起重要作用;VEGF与PTC转移密切相关。     

HIF-2α、VEGF与非小细胞肺癌中微血管生成及生物学转移的关系

应用免疫组化技术检测48例小细胞肺癌（NSCLC）中缺氧诱导因子-2α（HIF-2α）和血管内皮生长因子（VEGF）的表达,用CD34单克隆抗体标记血管内皮细胞并计数微血管密度（MVD）。结果48例NSCLC的HIF-2α和VEGF阳性表达率分别为72．9％和75％,与正常肺组织相比有统计学意义（P＜0．01）;HIF-2α和VEGF的表达呈正相关（r=0．721,P＜0．01）;HIF-2α或VEGF阳性NSCLC组织中MVD明显高于阴性组织;NSCLC转移者与未转移者的HIF-2α和VEGF比值有显著性差异（P＜0.01）。结论HIF-2α参与丁济导VEGF表达,促进NSCLC血管生成及转移的过程．可能在NSCLC早期形成和发展阶段有重要作用。     

胰腺癌组织中VEGF、PCNA与血管生成的关系及预后相关性

目的 探讨胰腺癌组织中VEGF、PCNA和血管生成的关系。方法 用免疫组织化学方法检测48例胰腺癌及癌旁组织、6例正常胰腺组织中VEGF、PCNA的表达和微血管密度（MVD）。结果 胰腺癌组织中VEGF、PCNA的阳性表达率分别为54．17％和77．5％，显著高于癌旁组织和正常组织的表达率（P〈0．01），胰腺癌组织中MVD显著高于癌旁组织和正常组织。VEGF表达与肿瘤大小和TNM分期有关（P=0．020，P=0．045），并且与MVD有相关性（r=0．294，P=0．043）。PCNA与临床病理因素无关。多元回归分析显示VEGF、PCNA和MVD都不是影响胰腺癌预后的独立因素。结论 血管生成在胰腺癌的发生、发展过程中起重要作用，抗肿瘤血管生成可能会提高胰腺癌的治疗效果。     

Overexpression of EphA2, MMP‐9, and MVD‐CD34 in hepatocellular carcinoma: Implications for tumor progression and prognosis

Aim: To investigate the expression of erythropoietin‐producing hepatocellular (Eph)A2 receptor, matrix metalloproteinase (MMP)‐9, and angiogenesis in hepatocellular carcinoma (HCC), in order to reveal their expression correlations with tumor invasion, metastasis, and prognosis. Methods: From January 2000 to June 2003, 129 specimens of resected tumors from the patients with HCC were obtained. Corresponding pericarcinomatous liver tissues were also obtained and selected as a control group. Expressions of EphA2, MMP‐9, and CD34 were detected with immunohistochemical staining. Microvascular density (MVD) was calculated with counting of CD34‐positive vascular endothelial cells. Results: The expressions of EphA2, MMP‐9, and MVD in the HCC tissues were significantly higher than those in the pericarcinomatous liver tissues (P < 0.01). Statistical analysis showed there were significant correlations between the expressions of EphA2, MMP‐9 and MVD in some classicclinicopathological parameters (i.e. tumor nodule, vein invasion, tumor, node, metastasis stages, extrahepatic metastasis; P < 0.05). The correlation between EphA2 and MMP‐9 expression was positive (r = 0.625, P = 0.011). Tumor MVD was closely associated with EphA2 (r = 0.281, P = 0.01) and MMP‐9 (r = 0.319, P < 0.01) expressions. In particular, EphA2, MMP‐9, and MVD expressions levels were found to be independent prognostic factors after HCC resection. Conclusions: Overexpressions of EphA2 and MMP‐9 relate to tumor progression, metastasis, and prognosis in HCC. The present study suggests that EphA2 is associated with key mediators of angiogenesis and invasion.     

骨桥蛋白下游信号分子在肝细胞癌中的表达

目的 通过肝癌组织芯片检测骨桥蛋白(OPN)下游的信号蛋白,以确定其在肝癌组织中的信号传导途径.方法构建肝细胞癌组织芯片,用免疫组织化学染色法检测并分析OPN及其相关分子、整合素αV、CD44v6、磷酸化黏着斑激酶(p-FAK)、p-Src、磷酸化细胞外信号调节激酶(p-ERK)、磷酸化蛋白激酶B(p-AKT)的表达水平及其关系.计数资料用卡方检验(或Fisher's确切概率法),各指标之间的相关性采用Spearman相关分析.结果OPN及其受体整合素αV、CD44v6和相关信号分子p-FAK、p-Src、Src、p-ERK、p-AKT在肝癌组织的表达水平均明显高于癌周正常肝组织(P＜0.05).FAK在肝癌和癌周组织中的表达差异无统计学意义(P＞0.05).OPN的表达与整合素oV(P＜0.01)、p-ERK(P＜0.01)、CD44v6(P＜0.05)密切相关,与p-FAK、p-Src、p-AKT无相关性(P＞0.05).但p-FAK(P＜0.05)、p-Src(P＜0.01)和p-AKT(P＜0.05)均与OPN受体整合素αV密切相关,p-FAK还与OPN的受体CD44v6密切相关(P＜0.01).结论 OPN通过其受体整合素αV、CD44v6激活下游的丝裂原活化蛋白激酶途径以促进肝癌转移.

Abstract：

Objective Osteopontin (OPN) has close relationship with metastasis in hepatocellular carcinoma but its downstream signal pathways have not been well defined in hepatocellular carcinoma. The object of this study is to identify the associated signal pathways in human HCC tissues. Methods The expressions of OPN, intergrin α V, CD44v6, P-FAK, FAK, P-Src, Src, P-ERK and P-AKT were assayed using TMA analysis. The relationship of OPN with P-ERK, P-Src and P-AKT were explored and the role in HCC metastasis was analysed. Results The expression levels of OPN, intergrin α V, CD44v6, P-FAK, P-Src, Src, P-ERK and P-AKT in HCC tissue were significantly higher than that in normal tissue (P ＜ 0.05). No significant difference was found between the expression levels of FAK in HCC tissue and normal tissue (P ＞0.05). OPN expression was significantly associated with Integrin α v (P ＜ 0.01 ), CD44V6 (P ＜ 0.01) and P-ERK (P ＜ 0.05) but not with P-Stc, P-FAK and P-AKT (P ＞ 0.05). The expressions of P-FAK (P ＜ 0.05), P-Src (P ＜ 0.01) and P-AKT (P ＜ 0.05) were significantly associated with Integrin α v and the P-FAK expression was also significantly associated with CD44V6 (P ＜ 0.01). Conclusion OPN promotes HCC metastasis though Integrin α v/CD44V6/MAPK pathway in human HCC.     

Role of transforming growth factor-beta1-smad signal transduction pathway in patients with hepatocellular carcinoma

AIM:To explore the role of transforming growth factor-beta1(TGF-β1)-smad signal transduction pathway inpatients with hepatocellular carcinoma.METHODS:Thirty-six hepatocellular carcinoma speci-mens were obtained from Qidong Liver Cancer Instituteand Department of Pathology of the Second AffiliatedHospital of Nanjing Medical University.All primary anti-bodies(polyclonal antibodies)to TGF-β1,type Ⅱ Trans-forming growth factor-beta receptor(TβR-Ⅱ),nuclear fac-tor-kappaB(NF-KB),CD34,smad4 and smad7,secondaryantibodies and immunohistochemical kit were purchasedfrom Zhongshan Biotechnology Limited Company(Bei-jing,China).The expressions of TGF-β1,TβR-Ⅱ,NF-kB,smad4 and smad7 proteins in 36 specimens ofhepatocellular carcinoma(HCC)and its adjacent tissuewere separately detected by immunohistochemistry toobserve the relationship between TGF-β1 and TβR-Ⅱ,between NF-kB and TGF-β1,between smad4 and smad7and between TGF-β1 or TβR-Ⅱ and microvessel density(MVD).MVD was determined by labelling the vesselendothelial cells with CD34.RESULTS:The expression of TGF-β1,smad7 and MVDwas higher in HCC tissue than in adjacent HCC tissue(P<0.01,P<0.05,P<0.01 respectively).The expressionof TβR-Ⅱ and smad4 was lower in HCC tissue than in its adjacent tissue(P<0.01,P<0.05 respectively).Theexpression of TGF-β1 protein and NF-kB protein wasconsistent in HCC tissue.The expression of TGF-β1 andMVD was also consistent in HCC tissue.The expressionof TβR-Ⅱ was negatively correlated with that of MVD inHCC tissue.CONCLUSION:The expressions of TGF-β1,TβR-Ⅱ,NF-kB,smad4 and smad7 in HCC tissue,which are ma-jor up and down stream factors of TGF-β1-smad signaltransduction pathway,are abnormal.These factors areclosely related with MVD and may play an important rolein HCC angiogenesis.The inhibitory action of TGF-β1 isweakened in hepatic carcinoma cells because of abnor-mality of TGF-β1 receptors(such as TβR-Ⅱ)and postre-ceptors(such as smad4 and smad7).NF-kB may causeactivation and production of TGF-β1.     

Microvessel density of malignant and benign hepatic lesions and MRI evaluation

AIM. To study the difference of microvessel density (MVD) between malignant and benign hepatic lesions and study the relationship between MVD and dynamic enhanced magnetic resonance imaging (MRI) for evaluation of microvessels within malignant and benign hepatic lesions. METHODS: A total of 265 specimens of hepatocellular carcinoma (HCC), 122 cirrhosis tissues and 22 hepatic benign lesions were enrolled for MVD by immunohistochemistry on tissue microarray, of which 49 underwent MRI examination before surgery, then contrast-to-noise ratios (CNR) and enhancement index (EI) in all the phases were calculated. Pearson correlation was performed for correlation analysis between CNR, EI and MVD. RESULTS: MVD of HCC was 22.7&#177;15.8 (mean&#177;SD), which was obviously higher than that of cirrhosis tissue (8.3&#177;7.6, P&lt;0.01), but was not statistically different from that of benign lesions (31.3&#177;22.7, P&gt;0.05). Among HCC, MVD of gradesⅠ-Ⅱ was 29.9&#177;18.6, which was much higher than those ofgrade Ⅲ (22.2&#177;18.2, P&lt;0.01) and gradeⅣ (22.9&#177;19.0, P&lt;0.01). MVD of HCC (P=0.018) and of benign lesions (P=0.014) were both correlative with CNR in arterial phase. CONCLUSION: Neoangiogenesis is an important feature for malignant tumor, and MVD may act as a biological marker in differentiating malignant from benign hepatic lesions. Dynamic enhanced MRI, especially image in arterial phase, may act as an MVD evaluation criterion for malignant and benign hepatic lesions.     

Osteopontin,a single marker for predicting the prognosis of patients with tumor‐node‐metastasis stage I hepatocellular carcinoma after surgical resection

Background and Aim: Osteopontin (OPN) has been linked to clinical outcomes in several solid tumors. However, it has not been fully evaluated whether OPN could be used as a single marker for the prognosis of patients with hepatocellular carcinoma (HCC), particularly in patients of the tumor‐node‐metastasis (TNM) stage I. Methods: A total of 151 patients with HCC who underwent surgical resection were enrolled, including 112 patients of the TNM stage I. OPN expression was evaluated using immunohistochemistry in the tissue microarrays derived from these patients. Immunoreactivity was classified according to the percentage and intensity of staining: negative (?), weak (+) and strong (++). The impact of OPN expression on survival of patients was analyzed. Results: In total, 65.6% (99 of 151) of HCC tissues expressed OPN. Overall survival in patients of OPN (?) group was significantly higher than those of OPN (+) or OPN (++) group (P = 0.049 and P = 0.001). Interestingly, in patients of the TNM stage I, OPN expression was correlated with the early recurrence after surgical resection (P = 0.001). Multivariate analysis showed that OPN expression was an independent prognostic factor for overall survival and disease‐free survival in patients with the TNM stage I HCC (hazard ratio, 2.272, P = 0.014 and 1.982, P = 0.037). Conclusions: These results suggest that OPN is commonly expressed in HCC and is a useful marker for predicting the prognosis of patients with the TNM stage I HCC, contributing to determining which individual patient needs adjuvant therapy to prevent the early recurrence after surgical resection.     

骨桥蛋白、血管内皮生长因子-A和乏氧诱导因子-1α在肝细胞癌中的表达及意义

目的探讨骨桥蛋白（OPN）、血管内皮生长因子-A（VEGF—A）和乏氧诱导因子（HIF）-1α在肝细胞癌（HCC）中的表达及临床意义。方法采用免疫组织化学法检测肝癌组（90例）、癌旁肝硬化组（20例）及正常组（15例）中OPN、VEGF—A和HIF-1α的表达。结果OPN、VEGF—A和HIF-1α在HCC组呈明显高表达,阳性率分别为76．67％（69／90）、71．11％（64／90）和73．33％（66／90）。其中OPN、VEGF—A显著高于癌旁肝硬化组及正常组,HIF-1α在HCC组的表达与肝硬化组无差异,与正常组有显著差异。OPN、VEGF—A、HIF—1α的表达与肝癌的临床病理特征如癌栓形成、包膜完整性、肿瘤分化和分期、肿瘤转移等有相关性（P〈0．05）,三者之间表达呈正相关性（P〈0．05）。结论OPN、VEGF—A、HIF-1α在肝癌中高表达,联合检测有助于判断肝癌的生物学特征,为肝癌的分子靶向治疗提供重要的可能靶点。     

Expression and significance of new inhibitor of apoptosis protein survivin in hepatocellular carcinoma

AM: To investigate expression and significance of inhibitor of apoptosis protein survivin in hepatocellular carcinoma (HCC). METHODS: The expression of survivin and vascular endothelial growth factor (VEGF) was investigated in 38 cases of HCC tissues and 38 liver cirrhosis tissues by immunohistochemistry and Western blot. The relationship between the expression of survivin and clinicopathological factors of HCC was analyzed. RESULTS: Survivin protein was detected in 23 (60.5%) of 38 HCCs and 3 (7.9%) of 38 liver cirrhosis tissues. In 23 cases of HCC which expressed survivin, the expression of VEGF was positive in 18 cases and slight positive or negative in 5 cases. While in 15 cases of HCC which did not express survivin, 12 cases did not express or slightly expressed, and 3 cases expressed VEGF. In liver cirrhosis tissues, the expression of VEGF was as follows: 24 cases were negative, 10 cases were weak positive and 4 cases were strong positive. The expression of survivin was coincident with the expression of VEGF in HCC (P<0.01). The expression of survivin in HCC had no relationship with the patients' age, gender, tumor size and differentiation level of HCC, while it was related to the metastasis of HCC. The protein quantitative analysis by Western blot also showed that overexpression of survivin in HCC was closely correlated to the expression of VEGF (P<0.01). Furthermore, stronger expression of survivin and VEGF was also found in patients with metastasis rather than in those with no metastasis (P<0.01). CONCLUSION: Survivin plays a pivotal role in the metastasis of HCC, and it has some correlation with tumorigenesis. The expression of survivin in the primary lesion is very useful as an indicator for metastasis and prognosis of HCC. It could become a new target of gene therapy of HCC.     

结肠癌COX-2mRNA表达与微血管密度关系的临床意义

目的通过观察结肠癌组织COX-2mRNA及CD34的表达，结合结肠癌组织中微血管密度（microvessel density，MVD）所见，探讨COX-2与肿瘤血管形成及病理特征的关系，为结肠癌生物治疗提供理论基础。方法 选择62例结肠癌、22例结肠腺瘤和22例正常结肠黏膜标本，采用原位分子杂交法检测COX-2mRNA并用MaxVisionTM快捷免疫组化法检测CD34表达，光镜下记数MVD。结果结肠癌、结肠腺瘤组织COX-2mRNA的阳性率明显高于正常黏膜，且有统计学意义（74．19％：36．36％，P=0．001；72．73％：36．36％，P=0．015）；结肠癌组平均MVD值高于腺瘤组和正常组，三组比较，有统计学意义（F=19．628，P=0．000）。在62例结肠癌组织中，高分化组COX-2mRNA阳性率高于低分化组（X^2=4．215、P=0．040）；进展期癌组的MVD高于早期癌组（t=3．079，P：0．003）；淋巴结有转移组MVD高于无转移组（t=3．180，P=0．002）；有血管侵犯组高于无血管侵犯组（t=2．093，P=0．041）；COX-2mRNA阳性组MVD高于阴性组，COX-2mRNA高表达组MVD高于低表达组，但差异均无统计学意义（P〉0．05）。结论MVD记数可作为判断肿瘤预后的有效评价指标，COX-2与肿瘤细胞的增殖和凋亡密切相关，与肿瘤血管生成无直接相关性。     

Relationship between inducible nitric oxide synthase expression and angiogenesis in primary gallbladder carcinoma tissue

AIM:To explore the relationship between angiogenesis and biological behaviors of primary gallbladder carcinoma (PGBC),the relationship between the expression of inducible nitric oxide synthase (iNOS) and biological behaviors of PGBC and its relationship with the expression of iNOS and angiogenesis of PGBC.METHODS: The expression of iNOS and micro-vessel density (MVD) were assessed by immunohistochemical method and image analysis system in 40 specimens of PGBC and in 8 specimens of normal gallbladder. The immunostaining results and related clinicopathologic materials were analyzed by statistical methods.RESULTS: MVD in PGBC was significantly higher than that in normal gallbladder tissue (46&#177;14 vs 14&#177;6, P&lt;0.05), and was not related with age, gender, tumor size and histological type. MVD of poorly and undifferentiated tumor tissues was higher than that of moderately-differentiated and well-differentiated tumor tissues (52&#177;9 vs43&#177;9 vs33&#177;6, P&lt;0.01).MVD of Nevin IV and V stages was higher than that of NevinI, Ⅱ and Ⅲ stages (52&#177;8 vs 37&#177;13, P&lt;0.01). MVD of cases with lymphatic or liver metastasis was significantly higher than that without liver metastasis (55&#177;6 vs 42&#177;10, P&lt;0.05) or lymphatic metastasis (53&#177;8 vs38&#177;8, P&lt;0.01). The positive level index (PLI) of iNOS in PGBC was 0.435&#177;0.134, and was not related with age, gender, tumor size, histological type,differentiation and clinical stage of PGBC. The PLI of iNOS in cases with lymphatic metastasis was higher than that without lymphatic metastasis (0.573&#177;0.078 vs 0.367&#177;0.064,P&lt;0.01). The PLI of iNOS in cases with liver metastasis was higher than that without liver metastasis (0.533&#177;0.067 vs0.424&#177;0.084, P&lt;0.05). There was a significant correlation between PLI of iNOS and MVD in PGBC (P&lt;0.05).CONCLUSION:Angiogenesis of PGBC is significantly related to the biological behaviors of PGBC. The expression of iNOS is related to the biological behaviors of PGBC. The detection of MVD and the expression of iNOS in PGBC can be used as parameters to determine the degree of malignancy and prognosis.