Human lymphoblastoid cells produce extracellular matrix-degrading enzymes and induce endothelial cell proliferation, migration, morphogenesis, and angiogenesis

Human lymphoproliferative diseases can be hypothesized to invade locally and to metastatize via mechanisms similar to those developed by a variety of solid tumors, i.e., the secretion of extracellular matrix-degrading enzymes and stimulation of angiogenesis. To assess this hypothesis, Namalwa, Raji, and Daudi cell lines (Burkitt’s lymphoma), LIK and SB cell lines (B-cell lymphoblastic leukemia), CEM and Jurkat cell lines (T-cell lymphoblastic leukemia), and U266 cell line (multiple myeloma) were evaluated for their capacity to produce matrix metalloproteinase-2 and -9, and urokinase-type plasminogen activator. These cell lines were also assessed for their ability: (1) to produce the angiogenic basic fibroblast growth factor and vascular endothelial growth factor; (2) to induce an angiogenic phenotype in cultured endothelial cells, represented by cell proliferation, chemotaxis, and morphogensis; (3) to stimulate angiogenesis in different in vivo experimental models. All cell lines expressed the mRNA for one or both metalloproteinases. Namalwa, Raji, LIK, SB, and U266 cells secreted the active form of both metalloproteinases, while Daudi, CEM, and Jurkat cells produced metalloproteinase-2 but not -9. In contrast, urokinase-type plasminogen activator was secreted only by SB cells. While Raji, LIK, SB, CEM, and Jurkat cells secreted both basic fibroblast growth factor and vascular endothelial growth factor, Daudi and U266 cells produced only the former, and Namalwa cells only the latter. Accordingly, the conditioned medium of all cell lines stimulated cell proliferation and/or chemotaxis in cultured endothelial cells, with the exception of that of Namalwa cells which was ineffective. The conditioned medium of CEM and Jurkat cells induced morphogenesis in cultured endothelial cells grown on a reconstituted basement membrane (Matrigel). Lastly, Namalwa, Raji, LIK, SB, U266, CEM, and Jurkat cells induced angiogenesis and mononuclear cell recruitment in the murine Matrigel sponge model and in a chick embryo chorioallantoic membrane assay. The extent of angiogenesis in both models was strictly correlated with the density of the mononuclear cell infiltrate. The results indicate that human lymphoproliferative disease cells possess both local and remote invasive ability via the secretion of matrix-degrading enzymes and the induction of angiogenesis which is fostered by host inflammatory cells and by an intervening ensemble of angiogenic factors.     

三氧化二砷对人脐静脉内皮细胞中蛋白激酶C活性和白细胞介素-15分泌、表达的影响及其临床意义

目的 通过研究三氧化二砷(As2O3)对人脐静脉内皮细胞(HUVEC)中蛋白激酶C(PKC)活性和白细胞介素(IL)-15表达和分泌的影响,探讨内皮细胞在As2O3抑制白血病或肿瘤中的作用。方法用不同浓度的As2O3作用于培养的HUVEC细胞,测定其生长情况、胞膜和胞浆PKC活性以及对IL-15基因表达和分泌的影响。结果0.25 μmol／L~50 μmol／L的.As2O3均抑制HUVEC细胞的增殖(P<0.01),随着药物浓度或作用时间的增加,细胞逐渐死亡。在As2O3作用细胞4 h测定胞浆和胞膜的PKC活性,两者在不同浓度作用下活性水平均得到提升(P<0.01),但胞膜的活性上升幅度更大。在As2O3的测定浓度范围内,PKC的活性水平与As2O3的浓度呈正相关。1、5、16 μmol／L的As2O3导致IL-15基因表达和分泌水平的增加(P<0.01),并呈浓度依赖性关系。结论As2O3可能是通过激活内皮细胞的PKC活性,促进IL-15或其他细胞因子的分泌,来抑制白血病或肿瘤发展的。     

Kidney Transplantation Substantially Improves Endothelial Progenitor Cell Dysfunction in Patients with End-Stage Renal Disease

Endothelial progenitor cells (EPC) are involved in endothelial repair and maintenance. Dysfunction of EPC may contribute to accelerated arteriosclerosis in chronic kidney disease. Kidney transplantation (KTx) improves both survival and endothelial function of dialysis patients. In a prospective study, we tested to which extent KTx changes EPC biology. We studied number and function (migratory activity, adhesion to extracellular matrix proteins and to mature endothelial cells [EC]) of EPC in 20 patients during dialysis and 3, 6, 9 and 12 months after KTx. Twenty-two healthy volunteers served as matched controls. Circulating precursor populations were measured by flow cytometric analysis. Cytokines relevant for EPC mobilization were monitored. Compared to the dialysis state, KTx increased the migration of EPC to approximately 2-fold. Adhesion to fibronectin and to collagen type IV was significantly increased after KTx. An improved adhesion rate of EPC to mature EC was observed. The number of EPC decreased. The amount of precursor populations showed no difference compared to the pretransplant state. Our study shows an improved function of EPC after KTx. This finding indicates an improved potential for endothelial repair which in turn may contribute to enhanced endothelial function and reduced cardiovascular morbidity after KTx.     

妊娠肝内胆汁淤积症患者胎盘组织中血管内皮生长因子的表达

目的观察血管内皮生长因子(VEGF)在妊娠肝内胆汁淤积症(ICP)胎盘中的表达.方法采用免疫组织化学方法,检测25例正常妊娠胎盘(对照组)和25例ICP胎盘(ICP组)中VEGF的水平.结果 VEGF在正常妊娠胎盘和ICP胎盘中分布基本一致,分布在滋养细胞、血管及绒毛间质.在ICP组VEGF为轻度表达占76%,中度表达占24%,无重度表达;而在对照组VEGF轻度表达占24%,中度表达占44%,重度表达占32%.ICP组中VEGF表达明显低于对照组(P＜0.01).结论 VEGF在胎盘中主要由绒毛滋养细胞分泌,ICP胎盘中VEGF的减少可能与胎盘血管生成减少及胎盘滋养叶细胞浸入异常有关,在ICP发病中占有一定的地位.     

阶跃切应力对内皮细胞释放前列环素的影响

采用流室定量研究阶跃切应力对内皮细胞释放前列环素（ｐＧＩ＿２）的影响。对流室中的内皮细胞连续１００分钟施加１、１０、２０ｄｙｎｅ／ｃｍ￣２的阶跃剪切应力，用放免法测流室的８个时间段流出液的６－ｋｅｔｏ－ＰＧＦｉ＿（１ａ）。绘出切应力刺激内皮细胞释放ＰＧＩ＿２的曲线。结果表明：未施加切应力时，内皮细胞释放ＰＧＩ＿２的量在检测限以下，施加切应力后，ＰＧＩ＿２释放速率很快达极大值，且又在数分钟内减小．施加阶跃切应力越大，ＰＧＩ＿２释放速率曲线峰值越高，但衰减也越快，在８０～１００分钟时间内的前列环素释放速率平均值也越大。由此提示，在活体中血液流变学因素一血流切应力对血管内皮细胞释放前列环素有促进作用。     

明显强化孤立肺结节血流模式的临床价值

目的利用4层螺旋CT动态增强技术定量评价不同性质的明显强化孤立肺结节的血流模式并初步评价血管内皮生长因子(VEGF)表达阳性的孤立性肺腺癌血管生成与血流模式定量CT参数的相关性.方法 78例孤立明显强化肺结节(直径≤4 cm,68例恶性,10例活动性炎性),行多层螺旋CT(MSCT)动态增强(以4 ml/s的流率注入对比剂).记录孤立肺结节增强前后各时相的CT值并计算强化值、灌注值,结节-主动脉强化值比.灌注值等于时间-密度曲线最大斜率除以主动脉强化值.其中30例VEGF表达阳性的肺腺癌患者用免疫组织化学测定微血管密度(MVD)并标定VEGF,评价肺腺癌血流模式定量CT参数(强化值、灌注值、结节-主动脉强化值比及平均通过时间)与MVD的相关性.结果恶性结节强化值(35.79±10.76) HU与活动性炎性结节(39.76±4.59) HU差异无显著意义 (t=1.148 , P=0.255).恶性结节的结节-主动脉强化值比(14.27±4.37)%及灌注值(3.02±0.96)ml-1·min-1·kg-1均低于活动性炎性结节(18.51±2.71)%,(6.34±4.39)ml-1·min-1·kg-1 (t=2.978,P=0.004;t=5.590,P<0.0001).VEGF表达阳性的肺腺癌强化值(33.06±13.57)HU、结节-动脉强化值比(14.25±4.92)%及灌注值(2.97 ±0.56) ml-1·kg-1·min-1与MVD(70.15±20.03)条/视野,均呈正相关性(r=0.781, P<0.0001;r=0.688, P<0.0001;r=0.716, P<0.0001).平均通过时间(14.86±5.84)s与MVD无显著相关性(r=0.260, P=0.200).结论恶性与活动性炎性孤立肺结节血流模式不同,恶性结节通过结节-大动脉强化值比和灌注值可有效区别于活动性炎性结节,有助于两者鉴别诊断.肺腺癌强化值、结节-动脉强化值比及灌注值反映了VEGF表达阳性的肺腺癌的MVD.强化值、结节-动脉强化及灌注值可作为VEGF相关的肺腺癌血管生成的指标.     

肺腺癌VEGF-C、VEGF-D的表达与MVD、MLVD及淋巴转移的关系

目的探讨肺腺癌中VEGF-C、VEGF-D与微淋巴管密度MLVD（VEGFR-3）、微血管密度MVD （CD34）及淋巴结转移之间的关系。方法免疫组化检测48例肺腺癌组织中VEGF-C、VEGF-D、MLVD、MVD蛋白的表达。结果VEGF-C、VEGF-D蛋白阳性率分别为70．8％（34／48例）、58．3％（28／48例）,肿瘤周边部位显著高于肿瘤中心部位,具有统计学意义,其表达与肿瘤分化程度无关,与肿瘤的TNM分期有关,Ⅲ～Ⅳ期显著高于Ⅰ～Ⅱ期。在VEGF-C蛋白阳性组,MVD高于阴性组（P=0．016）,MLVD显著高于阴性组（P=0．006）,淋巴结转移（P=0．042）增多;而VEGF-D蛋白阳性组与阴性组相比MVD无显著差异（P=0．943）, MLVD高于阴性组（P〈0．01）,淋巴结转移（P=0．012）增加。结论VEGF-C的表达与肺腺癌血管生成及淋巴管生成和淋巴结转移关系密切,而VEGF-D的表达只与淋巴管生成和淋巴结转移关系密切,与血管生成无关。     

灌注液对糖尿病玻璃体切割手术中角膜内皮的影响

目的 评价不同灌注液对糖尿病玻璃体切割手术中角膜内皮细胞的影响。方法 对30例术中应用BSS或BSSPlus的糖尿病玻璃体切割手术眼的角膜内皮细胞密度和细胞面积变异系数进行前瞻性研究．结果 无囊膜组中应用BSSPlus的玻切手术眼角膜内皮细胞密度、细胞面积变异系数与应用BSS的手术眼之间存在明显差异，在有囊膜组中二者之间无明显差异。结论 对于囊膜不完整的糖尿病患者玻切手术中使用BSSPlus可减轻手术对角膜内皮细胞的损害。     

对急性冠状动脉综合征的新认识和治疗对策

近几年来，由于基础实验研究的迅速发展，对急性冠状动脉综合征(ACS)的认识和治疗取得了重大进展。作者就ACS分子生物学和细胞学基础、不稳定斑块的治疗对策、ACS的危险分层及治疗方法作一综述。     

老年高血压妇女内皮功能、血小板活化状态及雌激素的影响

目的：了解老年高血压妇女内皮功能和血小板活化状态，以及雌激素治疗的影响。方法：已接受降压治疗的78例老年高血压妇女随机分为两组，均继续服用降压药，A组加服尼尔雌醇，B组加服安慰剂，疗程为6个月。治疗前后观察血浆一氧化氮（NO）、血管性血友病因子（vWF)、P选择素（P－selectin)和纤维蛋白原（Fbg)浓度变化。24例健康体检者作为正常对照组。结果：高血压vWF,P-selectin和Fbg显著高于对照组，NO显著低于对照组；治疗后A组vWF和P-selectin显著降低，NO明显升高，与B组比较有显著性差异。结论：已接受降压治疗的老年高血压妇女仍存在内皮损害和血小板活化，尼尔雌醇治疗具有明确的有益作用。