VEGF和八因子相关抗原在人脑胶质瘤中的表达及意义

目的 通过检测人脑胶质瘤细胞中血管内皮生长因子（VEGF）和八因子相关抗原（Ⅷ-R Ag）的表达,探讨VEGF在脑胶质瘤发生和发展中的作用.方法 正常脑组织对照标本3例,取自脑外伤后内减压的病人.62例胶质瘤病人的手术标本,其中胶质母细胞瘤23例,间变胶质细胞瘤20例和星形胶质细胞瘤19例.采用VEGF多克隆抗体、兔抗人八因子相关抗原（Ⅷ-R Ag）行常规S-P免疫组化染色,观察并计算VEGF免疫活性指数和微血管密度值.结果 VEGF染色指数随着肿瘤恶性度的升高而增高;脑胶质瘤组织微血管密度随着肿瘤级别的升高而增高;脑肿瘤病理分级和VEGF（r=0.941）的表达及与MVD（r=0.907）之间呈正相关性.结论 VEGF可能通过促进肿瘤组织内血管生成,在人脑胶质瘤的形成与发展过程中起着重要作用,其表达水平对判断恶性程度及预后有指导意义.     

VEGF在恶性淋巴瘤的表达及与MVD的相关性研究

目的探讨血管内皮生长因子(VEGF)在恶性淋巴瘤的表达,及与微血管密度(MVD)的关系。方法采用免疫组化法对恶性淋巴瘤淋巴结病理组织进行免疫学分类,及测定VEGF及微血管密度。结果(1)恶性淋巴瘤组织VEGF高表达,微血管密度增高,与对照组比较差异显著(P<0.01);(2)VEGF主要表达于淋巴瘤淋巴结外带,于肿瘤细胞及其周围细胞胞浆、血管内皮细胞及微血管周围表达;⑶非霍奇金恶性淋巴瘤VEGF与MVD呈正相关(P<0.01)。结论恶性淋巴瘤形成时已有VEGF高水平表达及丰富的微血管形成;VEGF由瘤细胞及周围组织细胞分泌,沉积于血管内皮细胞周围;非霍奇金淋巴瘤VEGF的分泌与微血管的形成相关。     

血管内皮生长因子和微血管密度与非小细胞肺癌预后的关系

目的:探讨血管内皮生长因子(VEGF)和微血管密度(MVD)与非小细胞肺癌(NSCLC)预后的关系.方法:运用免疫组织化学方法检测43例NSCLC石蜡切片中MVD和VEGF的表达,采用Kaplan-meier法和COX模型(似然比检验)进行分析.结果:单因素生存分析显示MVD阳性、VEGF阳性表达的肺癌患者术后生存期缩短;多因素COX回归分析显示MVD是NSCLC患者预后的独立预测因子.MVD阳性NSCLC患者术后死亡的相对危险度是MVD阴性患者的34.247倍(似然比检验,P<0.01).结论:MVD可能是NSCLC预后较强的独立预测因子,MVD高则NSCLC患者预后不良.     

微血管密度和VEGF在膀胱癌中的表达及意义

目的探讨微血管密度(MVD)和血管内皮生长因子(VEGF)在膀胱癌组织中的表达及临床意义。方法应用免疫组化S-P法检测40例膀胱癌组织和10例正常膀胱组织中MVD和VEGF的表达。结果40例膀胱癌组织中MVD记数为MVD为(47.9±8.7)。其中Ⅰ级、Ⅱ级和Ⅲ级膀胱癌MVD分别为(36.2±7.5)、(44.4±9.7)和(54.9±7.0);VEGF阳性率72.5%(29/40),Ⅰ级、Ⅱ级和Ⅲ级阳性率分别为43.8%(7/16)、85.7%(12/14)和100%(10/10)。正常膀胱组织MVD(13.4±7.8),VEGF表达为0%。结论MVD值和VEGF的表达与膀胱癌的病理分级及临床分期有关,可以作为膀胱癌诊断和预后的有效指标之一。     

子宫内膜样腺癌中LVD和VEGF表达的关系及生物学意义

目的探讨子宫内膜样腺癌淋巴管密度（LVD）和VEGF表达的关系及生物学意义。方法应用免疫酶标组织化学二步法检测74例子宫内膜样腺癌、15例子宫内膜不典型增生、14例增殖期子宫内膜中D2-40和VEGF的表达,分析D2-40标记的LVD与VEGF表达的相关性及在不同病变组间表达的差异性。结果子宫内膜样腺癌组中VEGF阳性率和LVD高于增殖期和不典型增生子宫内膜组（P〈0.05）;浸润深度≥1/2肌层组VEGF阳性和LVD高于黏膜内癌组和浸润深度〈1/2肌层组（P均〈0.05）;淋巴结转移组VEGF阳性率和LVD高于无淋巴结转移组（P〈0.05）,且子宫内膜样腺癌VEGF阳性组的LVD高于阴性组（P〈0.05）;子宫内膜样腺癌的VEGF阳性率和LVD与肿瘤细胞分化程度无关。结论VEGF和LVD与子宫内膜样腺癌的生物学行为有关,VEGF可促进淋巴管生成。     

MVD、VEGF在前列腺癌组织中表达与CDFI血流检测及临床病理因素的研究

目的探讨前列腺癌组织微血管密度(MVD)、血管内皮生长因子(VEGF)表达与彩色多普勒超声(CDFI)血流检测及临床病理关系。方法对27例前列腺癌,29例前列腺增生症进行CDFI血流检测,将前列腺内动脉血流显示情况进行分级,计测阻力指数(RI);采用免疫组化(SP)法检测相对应标本内MVD、VEGF的表达。结果前列腺癌组织中MVD与血流分级和病理分化有良好相关性,随血流分级增大和病理分化程度减低呈上升趋势;VEGF表达强度与血流分级有关,与病理分化无明显关联;转移癌RI、MVD、VEGF高于无转移者。结论检测前列腺癌血流及MVD、VEGF表达,对评价其生物学行为具有重要意义。     

膀胱癌血管内皮生长因子、CD31表达与浸润的关系

目的：探讨血管内皮生长因子（VEGF）、CD31在膀胱癌组织中的表达及其与肿瘤浸润的关系。方法：采用免疫组化SP法，分析59例原发性膀胱移行细胞癌VEGF的表达情况，检测CD31以计算微血管计数（MVC）。结果：VEGF可在膀胱癌中表达，VEGF与膀胱癌的分期相关，并与MVC呈正相关（P〈0．05）。结论：VEGF能调节膀胱癌的血管形成，可作为判断膀胱癌生物学行为的指标之一。     

EBER、PTEN和VEGF在血管免疫母T细胞淋巴瘤中的表达及其临床病理学意义

目的:探讨血管免疫母细胞T细胞淋巴瘤(angioimmunoblastic T-cell lymphoma,AITL)组织中EB病毒(Epstein-Barr virus,EBV)、PTEN和VEGF三者表达间的关系,并分析三者与患者临床病理学参数间的关系。方法:采用原位杂交法检测21例AITL组织中EBV编码的小RNA(EBER)的表达情况,免疫组织化学En Vision两步法检测21例AITL和20例淋巴结反应性增生组织中PTEN和VEGF的表达情况,分析三者表达间的相关性及其与患者临床病理学参数间的关系。结果:21例血管免疫母细胞T细胞淋巴瘤中EBER表达阳性率为61.9%;PTEN和VEGF在AITL和淋巴结反应性增生组织中的表达差异均具有显著性(P0.05);EBER与PTEN表达呈负相关(P0.05)。血管免疫母细胞T细胞淋巴瘤中,男性患者和进展期组EBER阳性表达率分别为80%和78.6%,明显高于女性患者和非进展期组患者(P0.05);在伴随B症状和进展期组PTEN阳性表达率分别为31.3%和21.4%,明显低于无B症状和非进展期组患者(P0.05)。生存分析显示,PTEN表达与患者总生存率呈负相关(P0.05)。结论:EBV感染和PTEN低表达可能预示血管免疫母细胞T细胞淋巴瘤恶化进展。EBV是否通过下调PTEN表达参与血管免疫母T细胞淋巴瘤的发生尚不清楚,需进一步研究探讨。     

HIF-1α和VEGF在子宫内膜异位症增生期内膜中的表达及意义

目的探讨缺氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)在子宫内膜异位症(EMs)增生期内膜组织中的表达及与血管生成的相关性。方法分别用SP法和Western blot方法检测内异症40例异位内膜、33例在位内膜及36例子宫肌瘤患者(对照组)增生期内膜组织中HIF-1α和VEGF蛋白表达,CD34标记新生血管,计算微血管密度(MVD)。结果①HIF-1α和VEGF蛋白表达均为异位内膜高于在位内膜、在位内膜高于对照组内膜,组间比较差异均有统计学意义;②异位内膜MVD高于在位内膜、而在位内膜高于对照组内膜,且任意2组之间比较差异均有统计学意义;③异位内膜组织中VEGF与HIF-1α、VEGF与MVD及HIF-1α与MVD之间均存在正相关关系,差异均有统计学意义。结论 VEGF和HIF-1α可能存在相互作用,通过促进内异症血管生成,在内异症的发生发展中发挥作用。     

Rapid G protein-regulated activation event involved in lymphocyte binding to high endothelial venules

The homing of blood borne lymphocytes into lymph nodes and Peyer's patches is mediated in part by recognition and binding to specialized high endothelial venules (HEV). Here we demonstrate that a rapid pertussis toxin-sensitive lymphocyte activation event can participate in lymphocyte recognition of HEV. In situ video microscopic analyses of lymphocyte interactions with HEV in exteriorized mouse Peyer's patches reveal that pertussis toxin has no effect on an initial "rolling" displayed by many lymphocytes, but inhibits an activation-dependent "sticking" event required for lymphocyte arrest. This is the first demonstration that physiologic lymphocyte-endothelial interactions can involve sequential rolling, activation, and activation-dependent arrest, previously shown only for neutrophils. The inhibitory effect of the toxin is dependent on its G protein-modifying ADP- ribosyltransferase activity and can be reversed by phorbol myristic acetate, which bypasses cell surface receptors to trigger activation- dependent adhesion. Lymphocyte sticking can occur within 1-3 s after initiation of rolling. We conclude that a rapid receptor-mediated activation event involving G protein signaling can trigger stable lymphocyte attachment to HEV in vivo, and may play a critical role in regulating lymphocyte homing.     

Tumor necrosis factor-dependent segmental control of MIG expression by high endothelial venules in inflamed lymph nodes regulates monocyte recruitment.

Monocytes recruited from the blood are key contributors to the nature of an immune response. While monocyte recruitment in a subset of immunopathologies has been well studied and largely attributed to the chemokine monocyte chemoattractant protein (MCP)-1, mechanisms mediating such recruitment to other sites of inflammation remain elusive. Here, we showed that localized inflammation resulted in an increased binding of monocytes to perifollicular high endothelial venules (HEVs) of lymph nodes draining a local inflammatory site. Quantitative PCR analyses revealed the upregulation of many chemokines in the inflamed lymph node, including MCP-1 and MIG. HEVs did not express detectable levels of MCP-1; however, a subset of HEVs in inflamed lymph nodes in wild-type (but not tumor necrosis factor [TNF] null mice) expressed MIG and this subset of HEVs preferentially supported monocyte binding. Expression of CXCR3, the receptor for MIG, was detected on a small subset of peripheral blood monocytes and on a significant percentage of recruited monocytes. Most importantly, in both ex vivo and in vivo assays, neutralizing anti-MIG antibodies blocked monocyte binding to inflamed lymph node HEVs. Together, these results suggest that the lymph node microenvironment can dictate the nature of molecules expressed on HEV subsets in a TNF-dependent fashion and that inflammation-induced MIG expression by HEVs can mediate monocyte recruitment.     

Regulation of leukocyte rolling and adhesion to high endothelial venules through the cytoplasmic domain of L-selectin

L-selectin (leukocyte adhesion molecule 1/MEL-14), a member of the selectin family of cell adhesion molecules, mediates leukocyte rolling and leukocyte adhesion to endothelium at sites of inflammation. In addition, L-selectin mediates the binding of lymphocytes to high endothelial venules (HEV) of peripheral lymph nodes. The strong amino acid sequence conservation of the cytoplasmic domain of L-selectin between humans and mice suggests an important role for this region. Deletion of the COOH-terminal 11 amino acids from the approximately 17 amino acid cytoplasmic domain of L-selectin eliminated binding of lymphocytes to HEV in the in vitro frozen section assay, and also abolished leukocyte rolling in vivo in exteriorized rat mesenteric venules, but did not alter the lectin activity of L-selectin. Pretreatment of cells with cytochalasin B, which disrupts actin microfilaments, also abolished adhesion without affecting carbohydrate recognition. Therefore, the cytoplasmic domain of L-selectin regulates leukocyte adhesion to endothelium independent of ligand recognition, by controlling cytoskeletal interactions and/or receptor avidity.     

Heparin promotes soluble VEGF receptor expression in human placental villi to impair endothelial VEGF signaling

Summary. Background: Severe preeclampsia is characterized by hypertension, renal injury and placental dysfunction. Prothrombotic disorders are discovered in 10–20% of women with preeclampsia, providing the rationale for prescribing low‐molecular‐weight heparin (LMWH) in future pregnancies. Heparin has diverse molecular actions and appears to reduce the recurrence risk of preeclampsia in women without prothrombotic disorders. The placenta‐derived anti‐angiogenic splice‐variant protein soluble vascular endothelial growth factor (VEGF) receptor‐1 (sFLT1) is strongly implicated in the pathogenesis of the underlying endothelial dysfunction. As the placental syncytiotrophoblast is the principal source of sFLT1, we tested the hypothesis that heparin suppresses placental sFLT1 secretion. Methods and Results: First trimester placental villi exposed to LMWH (0.25–25 IU mL^?1) in an in vitro explant model significantly increased the expression and release of sFLT1 by the syncytiotrophoblast into culture media, reducing phosphorylation of FLT1 and KDR receptors in cultured human umbilical vein endothelial cells. This response was significantly diminished in placental villi from healthy term pregnancies. Placental villi from severely preeclamptic pregnancies had a higher baseline sFLT1 release, compared with first trimester placental villi and did not respond to LMWH treatment. LMWH promoted villous cytotrophoblast proliferation (BrdU incorporation) and impaired syncytial fusion‐differentiation, causing syncytiotrophoblast apoptosis (by caspase 3&7 activity and TUNEL staining) and necrosis (ADP/ATP ratio). Conclusion: LMWH promotes sFLT1 synthesis and release from first trimester placental villi in a manner similar to that of severely preeclamptic placental villi, which antagonizes VEGF signaling in endothelial cells. These effects in part are mediated by an interaction between heparin and the cytotrophoblasts that regenerates the overlying syncytiotrophoblast responsible for sFLT1 secretion into the maternal blood.     

非小细胞肺癌组织中抑癌基因蛋白和血管内皮生长因子C的表达

目的探讨非小细胞肺癌(NSCLC)组织中Maspin蛋白与血管内皮生长因子C(VEGF-C)表达及其相关性。方法NSCLC组织石蜡标本100例,正常支气管黏膜及良性病变10例,采用免疫组化技术检测组织中Maspin蛋白与VEGF-C表达,结合临床病理,探讨两者与蛋白表达水平的意义。结果NSCLC组织与正常支气管黏膜及良性病变组织比较,Maspin蛋白阳性表达率较低,分别为51.0%vs 90.0%,VEGF-C表达率较高,分别为86.0%vs40.4%(均P<0.01)。结论在NSCLC组织中Maspin蛋白低表达,其转移能力增强可能与Maspin蛋白表达下调、缺失有关,有淋巴结转移比无淋巴结转移Maspin蛋白表达阳性率低;VEGF-C的高表达与组织分化和淋巴结转移有关,分化程度高和无淋巴结转移的组织VEGF-C的表达阳性率高。     

沉默信息调节因子2相关酶1表达与良性前列腺增生的相关性研究

目的通过研究沉默信息调节因子2相关酶1(SIRT1)在良性前列腺增生(BPH)组织中的表达和定位,以及与前列腺体积、年龄、血清前列腺特异抗原(PSA)的相关性,探讨衰老机制在BPH临床进展中的分子病理学机制。方法应用免疫组化法检测40例良性前列腺增生组织标本的SIRT1、细胞增殖指数(PI)以及凋亡抑制基因bcl-2的表达情况。记录所有患者的术前血清PSA水平、前列腺体积及年龄。分析SIRT1表达与上述各项指标以及PI、bcl-2表达的相关性。结果免疫组化法的结果显示SIRT1阳性表达率为55%(22/40),SIRT1表达与患者的年龄无显著相关。SIRT1阳性组平均血清PSA水平和前列腺体积均显著高于SIRT1阴性组(P值分别为0.031,0.044)。SIRT1阳性组bcl-2表达水平显著高于SIRT1阴性组(P=0.009),SIRT1的表达与PI无显著相关。结论 SIRT1表达与BPH患者前列腺体积增大、血清PSA水平升高以及bcl-2阳性表达密切相关。本研究结果提示SIRT1表达可能与BPH的临床进展密切相关。     

The role of chemokines in the microenvironmental control of T versus B cell arrest in Peyer's patch high endothelial venules

Chemokines have been hypothesized to contribute to the selectivity of lymphocyte trafficking not only as chemoattractants, but also by triggering integrin-dependent sticking (arrest) of circulating lymphocytes at venular sites of extravasation. We show that T cells roll on most Peyer's patch high endothelial venules (PP-HEVs), but preferentially arrest in segments displaying high levels of luminal secondary lymphoid tissue chemokine (SLC) (6Ckine, Exodus-2, thymus-derived chemotactic agent 4 [TCA-4]). This arrest is selectively inhibited by functional deletion (desensitization) of CC chemokine receptor 7 (CCR7), the receptor for SLC and for macrophage inflammatory protein (MIP)-3beta (EBV-induced molecule 1 ligand chemokine [ELC]), and does not occur in mutant DDD/1 mice that are deficient in these CCR7 ligands. In contrast, pertussis toxin-sensitive B cell sticking does not require SLC or MIP-3beta signaling, and occurs efficiently in SLC(low/-) HEV segments in wild-type mice, and in the SLC-negative HEVs of DDD/1 mice. Remarkably, sites of T and B cell firm adhesion are segregated in PPs, with HEVs supporting B cell accumulation concentrated in or near follicles, the target domain of most B cells entering PPs, whereas T cells preferentially accumulate in interfollicular HEVs. Our findings reveal a fundamental difference in signaling requirements for PP-HEV recognition by T and B cells, and describe an unexpected level of specialization of HEVs that may allow differential, segmental control of lymphocyte subset recruitment into functionally distinct lymphoid microenvironments in vivo.     

Triptolide涂层支架对猪冠脉内膜增殖及血管内皮生长因子、细胞间黏附分子-1表达的影响

目的 研究雷公藤内酯醇涂层支架(triptolide eluding stent,TES)置入猪冠状动脉后对血管内膜增生、炎性因子激活、血管损伤、术后再狭窄的影响及支架的安全性.方法 12只健康小型猪随机分为3组,分别植入雷公藤内酯醇涂层支架(100μg)、雷帕霉素洗脱支架(parter)、裸金属支架(每组12枚).每只猪冠状动脉前降支、回旋支、右冠状动脉各植入一枚支架,术后给予抗血小板药物,12周后复查冠状动脉造影(QCA)分析管腔变化、组织病理、血常规、生化结果,免疫组化榆测支架处血管VEGF,ICAM-1,α-actin的变化.结果 各组支架植入顺利,12周内无死亡,血管损伤积分无差异,管腔内均无血栓形成.裸支架组支架部位血管腔面积(3.76±0.61)mm2、最小管腔内径(2.15±0.18)mm小于TES组[(5.13±0.46)mm2,(2.65±0.21)mm]和DES组[(5.01±0.54)mm2,(2.65±0.25)mm,P＜0.01];裸支架内增生内膜面积及内膜增生程度裸支架组大于TES组和乐普支架组(P＜0.05),裸支架组支架部位血管内膜VEGF,ICAM-1,α-actin表达明显强于TES和乐普支架组.支架植入前后各组动物血细胞计数和肝肾功能无明显变化.结论 雷公藤内酯醇洗脱支架能抑制血管新生内膜增殖和炎性因子的表达,在支架植入12周后能有效地预防再狭窄.