“Real life” polycyclic aromatic hydrocarbon (PAH) mixtures modulate hCG,hPL and hPLGF levels and disrupt the physiological ratio of MMP-2 to MMP-9 and VEGF expression in human placenta cell lines

Using JEG-3 and BeWo cells, we examined the effect of “real life” mixtures of polycyclic aromatic hydrocarbons (PAHs), at doses reported in maternal blood (Mix I) and in placental tissue (Mix II), on human chorionic gonadotropin (hCG), placental lactogen (hPL) and placental growth factor (hPLGF) secretion, protein expression and immunolocalization. Additionally, the action of PAH mixtures on basal and hormone-stimulated matrix metalloproteinase-2 (MMP-2), MMP-9 and vascular endothelial growth factor (VEGF) protein expression was evaluated. Under basal conditions, the PAH mixtures increased hCG and decreased hPLGF levels in both cell lines, while hPL expression was stimulated in JEG-3 and inhibited in BeWo. There was no effect on the MMP-2/MMP-9 ratio or VEGF expression. In hormone-stimulated cells, PAH mixtures changed the MMP-2/MMP-9 ratio in JEG-3 cells in favor of MMP-9, while in BeWo MMP-2 was favored. The effect on VEGF expression was cell specific and dependent on the mixture. In hCG-treated cells, only Mix II inhibited VEGF expression in JEG-3 cells. Neither PAH mixtures affected this protein in BeWo cells. In hPL-treated cells, Mix I had a stimulatory effect in JEG-3 cells, while Mix II exerted an inhibitory effect in BeWo cells. In hPLGF-treated cells, Mix II decreased in JEG-3 cells, but in BeWo cells, both mixtures increased VEGF expression. Considering that the evaluated protein hormones play crucial roles in angiogenesis and neovascularization in the placenta, “real life” PAH mixtures by disrupting protein hormones levels, the MMP-2/MMP-9 ratio and VEGF expression can lead to insufficiency and many pregnancy-related disorders.     

A DNA vaccine expressing an optimized secreted FAPα induces enhanced anti-tumor activity by altering the tumor microenvironment in a murine model of breast cancer

Cancer-associated fibroblasts (CAFs), major components of the tumor microenvironment (TME), promote tumor growth and metastasis and inhibit the anti-tumor immune response. We previously constructed a DNA vaccine expressing human FAPα, which is highly expressed by CAFs, to target these cells in the TME, and observed limited anti-tumor effects in the 4T1 breast cancer model. When the treatment time was delayed until tumor nodes formed, the anti-tumor effect of the vaccine completely disappeared. In this study, to improve the safety and efficacy, we constructed a new FAPα-targeted vaccine containing only the extracellular domain of human FAPα with a tissue plasminogen activator signal sequence for enhanced antigen secretion and immunogenicity. The number of CAFs was more effectively reduced by CD8⁺ T cells induced by the new vaccine. This resulted in decreases in CCL2 and CXCL12 expression, leading to a significant decrease in the ratio of myeloid-derived suppressor cells in the TME. Moreover, when mice were treated after the establishment of tumors, the vaccine could still delay tumor growth. To facilitate the future application of the vaccine in clinical trials, we further optimized the gene codons and reduced the homology between the vaccine and the original sequence, which may be convenient for evaluating the vaccine distribution in the human body. These results indicated that the new FAPα-targeted vaccine expressing an optimized secreted human FAPα induced enhanced anti-tumor activity by reducing the number of FAPα⁺ CAFs and enhancing the recruitment of effector T cells in the 4T1 tumor model mice.     

VEGF在膀胱移行细胞癌中的表达及意义

目的：探讨膀胱移行细胞癌中血管内皮生长因子（VEGF）的表达及其与临床病理指标的关系。方法：应用免疫组化方法对40例膀胱移行细胞癌及15例正常膀胱组织中VEGF进行检测。结果：正常膀胱移行上皮均为阴性表达；膀胱移行细胞癌中VEGF阳性表达24例，表达率为60％（24／40），明显高于正常膀胱组织（P＜0.01），其中VEGF表达阳性率随膀胱癌病理分级、临床分期上升而升高，表达强度也随之增强（P＜0.01）。结论：VEGF表达对膀胱移行细胞癌生物学行为有重要影响，VEGF有可能可作为判断膀胱癌生物学行为、转移潜能及预后指标。     

A comparative study of angiogenic and cytokine responses after laparoscopic cholecystectomy performed with standard- and low-pressure pneumoperitoneum

Background  Surgical procedures enhance production of pro- and anti-inflammatory cytokines and angiogenic factors that play a pivotal role in the immunological response to surgical trauma and take part in the pathogenesis of tumor growth and adhesions formation. The purpose of the study was to access the influence of low-pressure CO₂ pneumoperitoneum on the inflammatory and angiogenic responses during the postoperative period after laparoscopy. Methods  The study group consisted of 40 patients, operated on due to cholelithiasis using standard-pressure (n = 20) and low-pressure (n = 20) CO₂ pneumoperitoneum. Serum concentration of interleukin (IL)-6, IL-8, IL-10, vascular endothelial growth factor (VEGF)-A, and endostatin were measured before and at 6, 24, and 48 h after surgery with commercially available enzyme-linked immunosorbent assay (ELISA). Results  Concentrations of IL-6 increased significantly after the operations in both groups. No differences were observed between the groups in regards to IL-6, IL-8, and IL-10 levels. Concentrations of VEGF-A measured at 6 and 48 h were significantly lower in patients who underwent laparoscopies performed with low-pressure pneumoperitoneum. No significant variations were observed in endostatin serum concentration. Concentrations of the studied parameters were not influenced by duration of surgery or by age, gender, or body mass index (BMI) of the patients. Conclusions  The results obtained in our study do not show any significant differences between studied operative procedures with regards to systemic inflammatory response. Changes in the concentrations of VEGF-A and endostatin observed in the studied population may suggest this technique is more favorable with regards to angiogenesis process intensity, along with all its consequences and implications.     

巨噬细胞移动抑制因子与鼻咽癌微血管生成和淋巴结转移的关系

[目的]研究巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)和血管内皮生长因子(vascular endothelial growth factor,VEGF)在鼻咽癌组织中的表达水平及其与肿瘤新生血管的关系,探讨细胞因子在鼻咽癌细胞侵袭转移过程中的作用.[方法]收集1999-2003年期间45例确诊未治的鼻咽原发癌活检组织标本,用LSAB免疫组化法检测鼻咽癌组织中MIF和VEGF表达,计数癌组织中新生血管热区的CD34阳性微血管密度(intratumoral microvessel density,IMD),并将各检测指标与患者的病理及临床资料相联系.[结果]鼻咽原发癌组织中,癌细胞MIF和VEGF的阳性表达率分别为78%(35/45)和71%(32/45),伴有淋巴结转移的癌组织中MIF和IMD水平均高于无淋巴结转移的癌组织,P=0.029,P=0.026,MIF阳性组的IMD计数为(35.1±13.3)/高倍视野,明显高于MIF阴性病例的(20.9±10.2)/高倍视野,P=0.003.以Schmincke型生长方式分布的癌细胞MIF表达水平(67.4%±35.2%)高于以Regaud型方式分布的癌细胞(32.9%±29.7%),P=0.007.癌细胞MIF与VEGF表达以及IMD计数均显示正相关关系,P＜0.05.但癌细胞VEGF的表达与患者性别、年龄、病理组织学分型以及临床分期无统计学显著意义.[结论]MIF在鼻咽癌细胞的淋巴结转移过程中可能具有重要的促进作用,并可能是影响肿瘤性微血管生成和癌细胞VEGF表达的重要因素.     

白介素-6、血管内皮生长因子、D-二聚体对CSDH的影响

目的探讨血肿局部炎症、假膜新血管生成、局部纤溶状况及其在CSDH发生、发展中的作用。进而探讨CSDH的发病机制，并为CSDH的治疗及预防复发提供理论依据。方法以78例CSDH患者作为病例组，20例健康人作为正常对照组。采用ELISA法测定患者血清及血肿液中VEGF及IL-6的含量。比较患者末梢静脉血及血肿液中四种因子的含量变化并与正常对照组比较。结果病例组血肿液FDP、d-dimer检测均为阳性，血液为阴性；正常对照组血液FDP、d-dimer检测均为阴性；病例组血清VEGF含量与正常对照组比较差异无统计学意义。血肿液中VEGF浓度高于血清中。病例组血清IL-6浓度与正常对照组差异无统计学意义，血肿液中IL-6浓度高于血清中。CSDH患者血肿液VEGF、IL-6水平没有相关性。结论CSDH患者血肿液局部纤溶亢进，局部VEGF分泌旺盛，新血管生成活跃，局部炎症活跃，可导致CSDH不断扩大而参与CSDH发病机制。抗炎治疗、抑制VEGF的生理作用、有选择的对病人施行促凝治疗，可成为部分CSDH病人保守治疗及预防复发的有效手段。     

外源性VEGF对自体移植脾组织血管再生的影响

目的：观察外源性VEGF对自体脾组织大网膜内移植后血管再生过程影响。方法：198只昆明种小白鼠随机分为脾切除自体脾组织大网膜内移植组及假手术组,将脾切除自体脾组织大网膜内移植后的小鼠再随机分为实验组（应用外源性VEGF组）和对照组（不用外源性VEGF组）。实验观察组小鼠分别于术后7、15、30d于尾静脉内注射VEGF,最后一次注射后1、7、14、30、60、90d各处死5只,取脾组织标本;对照组于术后观察上述对应时相点后各处死3只,取脾组织标本,进行墨汁灌注血管面密度测定。结果：术后7、15、30d注射VEGF后,较对照组的血管数量明显增多,墨汁灌注显示,实验观察组血管面密度较对照组明显增大。结论：外源性VEGF能够促进移植脾组织内血管生长,改善移植脾组织的血液循环,有利于移植脾组织再生,使其结构恢复更趋完善。     

奥曲肽对肝动脉栓塞治疗大鼠肝癌影响的实验研究

目的　观察奥曲肽(octreotide)降低肝动脉栓塞(HAE)促进肿瘤新生血管生长的作用以及抑瘤作用。方法　采用大鼠肝内种植Walker 256瘤株制作肝癌模型,分为对照组、Octreotide组、HAE组和HAE+octreotide组共四组。定期切取肿瘤标本测量肿瘤体积,检测标本中的血管内皮生长因子(VEGF)的表达及微血管密度(MVD)。结果　各干预组肿瘤体积均小于对照组(P<0.01), HAE+octreotide组肿瘤体积也小于 HAE组(P<0.05); HAE+octrotide组 VEGF表达低于HAE组(P<0.05);Octreotide组、HAE+octreotide组 MVD表达低于对照组(P<0.05)。结论　在对大鼠肝癌的治疗中,行肝动脉栓塞时联用奥曲肽,可降低单纯肝动脉栓塞引起的癌组织 VEGF高表达,减少肿瘤新生血管形成,加强其抑瘤作用。     

重组血管内皮细胞生长因子在大肠杆菌的高效表达

目的:使重组血管内皮细胞生长因子(VEGF)在大肠杆菌中得到高效表达.方法:通过构建表达重组VEGF的质粒PRL621/VEGF,并在大肠杆菌中高效表达.结果:表达量约占菌体总蛋白的40%.对形成包含体的表达产物进行变性,初步复性处理,得到重组人VEGF粗提液,鸡胚绒毛尿囊膜试验表明有促血管生长活性,N-端15个氨基酸序列分析结果,与天然VEGF蛋白质相应序列一致.