BPX-80离心式血泵中剪切力引发血管性血友病因子机械损伤的评价

目的 对BPX-80离心式血泵中剪切力引起血管性血友病因子（von Willebrand Factor,vWF）的损伤进行评价,判断其是否可以作为vWF损伤研究的参照泵。方法 按照ASTM标准搭建体外溶血测试平台,使用新鲜猪血在此平台对BPX-80离心式血泵进行8 h测试,然后评估每小时血液样本的溶血水平和vWF损伤,并与静态对照组进行比较。结果 在整个实验过程中,BPX-80的溶血指数保持稳定且较低;高分子量vWF多聚体有少量降解,与静置对照组相比无显著性差异;vWF抗原含量无明显变化,与静置对照组趋势基本一致。结论 BPX-80离心式血泵血液相容性良好,可以作为vWF损伤以及溶血评价实验的基准参照泵,从而为新型血泵的设计与优化提供参考。     

血小板反应素-1对猪胸导管内皮细胞增殖和凋亡的影响

目的探讨血小板反应素-1(thrombospondin-1,TSP-1)对淋巴管内皮细胞(lymphatic endothelial cells,LECs)增殖和凋亡的影响。方法取健康猪的胸导管内皮细胞进行分离、培养;应用凝血因子VIII,血管内皮生长因子受体-3(VEGFR-3)抗体联合对淋巴管内皮细胞进行鉴定;采用MTT法检测TSP-1对LEC的生长抑制作用;采用Hoechst33258荧光染色检测淋巴管内皮细胞凋亡。结果培养的淋巴管内皮细胞呈第VIII因子和VEGFR-3阳性反应,为典型淋巴管内皮细胞。MTT法证实,当浓度为0.8μg/ml￣2.0μg/ml时,TSP-1能明显抑制淋巴管内皮细胞的增殖,且抑制作用呈现剂量依赖关系。Hoechst33258证实,经TSP-1处理后淋巴管内皮细胞,可观察到其核周围有凋亡小体。结论TSP-1对淋巴管内皮细胞增殖有明显的剂量依赖性抑制作用,诱导淋巴管内皮细胞凋亡。     

不稳定性心绞痛患者血管性血友病因子与血管细胞间粘附分子水平的变化

目的：探讨老年不稳定性心绞痛（UA）患者血管性血友病因子（vWF）与可溶性血管细胞间粘附分子-1（sVCAM-1）水平变化及其与心肌缺血的关系。 方法： 采用酶联免疫吸附法（ELISA），检测了50例健康人和73例UA（自发性心绞痛27例，心肌梗死后心绞痛25例，恶化劳力性心绞痛21例）患者血浆vWF、sVCAM-1浓度的变化。 结果： ①UA患者血浆vWF水平、sVCAM-1浓度［（2.47±0.88）%、（1.92±0.51）%、（961±58）μg/L、（692±73）μg/L］，明显高于对照组［（572±58）μg/L、（0.96±0.14）%］（P＜0.01）；②心绞痛发作时vWF、sVCAM-1浓度增高较缓解后更明显（P＜0.01）；③心绞痛发作时和缓解后，sVCAM-1与vWF呈正相关（r=0.785，r=0.674，P＜0.01 ）；④不同类型的心绞痛发作时和缓解后sVCAM-1、vWF浓度差异亦具有显著性（P＜0.01）；⑤自发性心绞痛患者vWF、sVCAM-1增高较心肌梗死后和恶化劳力性心绞痛更明显。 结论： 急性心肌缺血与vWF和sVCAM-1异常有一定关系。     

流体切应力对血管内皮细胞粘附分子ICAM-1的影响

目的 :探讨血液流动产生的切应力对白细胞 血管内皮细胞 (VEC)相互粘附所产生的影响及其在炎症、免疫反应、动脉粥样硬化发生发展中所起的重要作用。方法 :将已汇合的人脐静脉内皮细胞玻片放于平板流动腔中 ,以层流不同切应力 0 .6、1.2、2 .4Pa剪切不同时间 (2、8、12h) ,用免疫细胞化学方法检测VEC表面ICAM 1表达 ,以显微镜下计数阳性细胞率表示。结果 :稳层流切应力能上调HUVECICAM 1表达 ,切应力是内皮细胞粘附分子表达的调节因素之一。     

循证血管发生因子PD-ECGF、VEGF及抑制因子TSP-1在子宫内膜异位症发病因素中的作用

目的 :循证血小板衍化内皮细胞生长因子PD ECGF、血管内皮细胞生长因子VEGF、血小板反应素TSP 1在子宫内膜异位症 (EM )发病因素中的作用。方法 :3 0例需行手术治疗的EM患者 ,取其在位和异位内膜作免疫组化SP法检测PD ECGF、VEGF ,原位杂交法检测TSP 1的表达 ,以 3 0例非EM患者的子宫内膜作为对照 ,进行比较分析。结果 :异位内膜PD ECGF表达升高 ,EM患者子宫内膜TSP 1表达较对照组降低。结论 :PD ECGF参与了EM血管发生 ,EM患者子宫内膜抑制血管发生能力降低 ,使异位的内膜易于种植生长可能是EM发病的因素之一     

流体切应力和Ox—LDL地内皮细胞表面ICAM—1表达的影响

目的研究动脉粥样硬化(AS)的危险因素高脂血症、流体切应力及二者共同作用对血管内皮细胞细胞间粘附分子-1(ICAM-1)表达的影响。方法以人脐静脉内皮细胞(HUVECs)为研究对象,应用免疫组化ACB方法,观测Ox-LDL、流体切应力及二者共同作用对HUVECs的粘附分子ICAM-1表达的影响。结果与对照组相比,(1)Ox-LDL显著增加HUVECs表面ICAM-1表达;(2)流体切应力使HUVECs表面ICAM-1表达增加,但ICAM-1的表达随时间依赖性增加的趋势不明显;(3)共同作用后,HUVECsICAM-1的表达较基础表达显著增加。结论Ox-LDL、流体切应力的改变及二者共同作用显著增加H-UVECs表面ICAM-1表达。     

细胞外基质蛋白1在肝癌中的表达及意义

目的：探讨细胞外基质蛋白1（extracellular matrix protein1,ECMl）在肝癌中的表达及意义。方法：应用免疫组织化学EnVision法,检测肝脏细胞株（正常肝细胞株LO2、肝癌细胞株HepG2）和肝脏组织（正常肝组织和肝癌组织）中ECM1的表达,并比较正常肝组织和肝癌组织之间的ECM1表达差异。结果：ECM1的表达如下：正常肝细胞株LO2（-）,肝癌细胞株HepG2（＋）;正常肝组织阳性率20％（4／20）,肝癌组织阳性率85．4％（70／82）。统计分析表明,对ECM1的表达,肝癌组织明显高于正常组织（P〈0．01）。结论：ECM1在肝癌组织中过表达。     

流体切应力对人内皮细胞迁移和整合素基因表达的影响

目的研究流体切应力空间梯度和均匀切应力对内皮细胞（ECs）迁移和整合素基因表达的影响，为阐明应力诱导血管重建的分子机制提供一些实验依据。方法将脐静脉内皮细胞置于平行平板流动腔系统，分别施加11dyn／cm^2的均匀切应力（矩形组）和5～14dyn／cm^2的切应力梯度（梯度组）为受力组，以静态条件下培养的ECs为对照组（静止组）。Transwell方法检测切应力对ECs迁移能力的影响；半定量RT-PCR测定ECs整合素α3β1、α5β1mRNA3、6和12h的表达情况。结果①同静止组相比，切应力梯度组ECs3h的迁移（88±4 vs 23±3，P〈0．01，n=3）能力显著提高；而均匀切应力组ECs3h迁移（21±3VS23±3）同静止组相比，差异无统计学意义（P〉0．05，n=3）；②ECs受切应力梯度作用3h即可诱导ECs整合素α3β1、α5β1mRNA表达（P〈0．01，P〈0．05）；均匀切应力于6h激活ECs整合素理，亚型表达（P〈0．01），α3 、β1亚型表达延迟到12h激活（P〈0．01）。结论切应力通过上调整合素基因表达促进了ECs迁移，提示整合素信号通路参与了切应力条件下ECs迁移过程的信号转导。     

Platelets, inflammatory cells, von Willebrand factor, syndecan-1, fibrin, fibronectin, and bacteria co-localize in the liver thrombi of Bacillus anthracis-infected mice

Vascular dysfunction and thrombosis have been described in association with anthrax infection in humans and animals but the mechanisms of these dysfunctions, as well as the components involved in thrombi formation are poorly understood. Immunofluorescent microscopy was used to define the composition of thrombi in the liver of mice challenged with the Bacillus anthracis Sterne spores. Lethal infection with the toxigenic Sterne strain, in contrast to the non-lethal, non-toxigenic delta-Sterne strain, demonstrated time-dependent increase in the number of vegetative bacteria inside the liver sinusoids and central vein. Massive appearance of thrombi typically occluding the lumen of the vessels coincided with the sudden death of infected animals. Bacterial chains in the thrombi were stained positive for syndecan-1 (SDC-1), fibronectin, and were surrounded by fibrin polymers, GPIIb-positive platelets, von Willebrand Factor (vWF), CD45-positive leukocytes, and massive amount of shed SDC-1. Experiments with human umbilical vein endothelial cells (HUVECs) demonstrated the active role of the host response to the secreted pathogenic factors of bacteria during the onset of the pro-thrombotic condition. The bacterial culture supernatants, as well as the isolated proteins (the pore-forming toxin anthrolysin O and phospholipase C) induced release of vWF, while anthrolysin O, sphingomyelinase and edema toxin induced release of thrombin from HUVECs and polymerization of fibrin in the presence of human plasma. Conclusion: Our findings suggest that activation of endothelium in response to infection can contribute to the formation of occlusive thrombi consisting of aggregated bacteria, vWF, shed SDC-1, fibrin, activated platelets, fibronectin and leukocytes.     

Adhesion Molecules ICAM-1 and PECAM-1 as Potential Biomarkers of Central Nervous System Damage in Women Breast Cancer Survivors

Breast cancer (BC) is the most common tumor in women worldwide with high mortality rates. Surgical methods followed by radio–chemotherapy are used to treat these tumors. Such treatment can lead to various side effects, including neurological complications. The development of a reliable biomarker to predict the onset of CNS complications could improve clinical outcomes. In the current study, ICAM-1 and PECAM-1 serum levels were measured as potential biomarkers in 45 female patients in a long-term follow-up period after breast cancer treatment, and compared to 25 age-matched female healthy volunteers. Serum levels of both biomarkers, ICAM-1 and PECAM-1 were significantly higher in patients after breast cancer treatment and could be associated with cognitive dysfunction, depression, and vestibulocerebellar ataxia. In conclusion, our results provide a first hint that elevated serum levels of ICAM-1 and PECAM-1 could serve as early predictive biomarkers in breast cancer survivors that might help to improve the management of these patients.     

缺氧应激反应中活性氧与缺氧诱导因子-1的相互作用

在缺氧条件下,线粒体会产生大量的活性氧（ reactive oxygen species, ROS）,随即大量的活性氧将会诱导机体产生缺氧应激反应。缺氧诱导因子1（hypoxia inducible factor 1, HIF?1）是缺氧应激反应中的中枢调控因子。有研究表明, ROS主要通过抑制脯氨酸羟化酶家族（ prolyl hydroxylases, PHDs）的活性来抑制HIF?1α的泛素化降解,从而稳定HIF?1。而HIF?1蛋白水平的升高有助于机体应对缺氧微环境。最近研究还发现, REDD?1可以通过ROS对HIF?1产生负调控作用,从而抑制肿瘤的形成;秀丽线虫呼吸突变体的产生会导致ROS水平增加,从而增强HIF?1的活性,最终延长的线虫寿命;以及ROS、 HIF?1促进自噬的产生。因此,了解缺氧条件下ROS与HIF?1之间的相互作用关系,对于今后肿瘤、衰老和自噬的研究具有重要意义。     

细胞因子调节人肝癌细胞ICAM-1分子的表达及其对CD3AK杀伤功能的影响

应用基因重组人干扰素α（IFN-α）、干扰素γ（IFN-γ）和肿瘤坏死因子（TNF）体外处理人肝癌细胞系H-7402,ABC-CELISA法检测各处理组和对照组细胞表面细胞间粘附分子-1（ICAM-1）的表达水平,用 LDH释放法观察CD3单克隆抗体活化的杀伤细胞（CD3AK）对各处理条件下H-7402细胞的杀伤活性。结果表明:经TNF,IFN-α处理的肿瘤细胞ICAM-1表达水平较对照组显著增高,同时对CD3AK细胞的杀伤敏感性亦显著增强,IFN-γ虽能够提高靶细胞ICAM-1分子的表达,但CD3AK的杀伤活性并未因此增强。抗ICAM-1和抗白细胞功能相关抗原LFA-1单抗能够有效地降低CD3AK对靶细胞的杀伤作用,并且能够阻断IFN-α、TNF对CD3AK细胞杀伤效应的促进作用。提示ICAM-1/LFA-1参与了CD3AK粘附杀伤肿瘤细胞过程,TNF、IFN-α的促杀伤效应与肿瘤细胞表面ICAM-1分子的表达水平升高有关。     

血清TK1、CD147、VEGF、CYFRA21-1和CEA联合检测对非小细胞肺癌的诊断价值研究

目的探究血清胸苷激酶1(TK1)、细胞外基质金属蛋白酶诱导因子(CD147)、血管内皮细胞生长因子(VEGF)联合细胞角蛋白19片段(cytokeratin-19-fragment,CYFRA21-1)和癌胚抗原(carcinoembryonic antigen,CEA)对非小细胞肺癌(NSCLC)的诊断、风险评估价值,及五者与NSCLC临床病理特征之间的关系。方法收集2016年5月至2018年4月期间入我院就诊的NSCLC患者120例作为病例组,同期收取我院收治的患有肺部良性疾病患者120例作为对照组。结果与对照组相比,NSCLC患者血清TK1、CD147、VEGF、CYFRA21-1和CEA水平均增高,且差异具有统计学意义(P<0.05)。血清TK1、CD147和VEGF与NSCLC患者淋巴结转移及TNM分期显著相关(P<0.05),且三者的水平在淋巴结转移或Ⅲ~Ⅳ期患者中显著高于未发生淋巴结转移或Ⅰ~Ⅱ期患者(P<0.05)。血清TK1、CD147、VEGF、CYFRA21-1和CEA在区分NSCLC与肺部良性疾病的灵敏度/特异性分别为:91.2%/82.2%、81.0%/90.9%、79.1%/80.4%、68.8%/61.4%和56.1%/53.7%;对应的临界值分别为:3.2pmol/L、5.4ng/L、88.7pg/mL、9.2ng/mL和4.9ng/mL;联合血清TK1、CD147、VEGF、CYFRA21-1和CEA在区分NSCLC与肺部良性疾病的AUC为0.949(95%CI:0.905~0.998,P<0.001),在"并联"时,约登指数为最大值时的灵敏度和特异性分别为93.6%和86.4%;在"串联"时,约登指数为最大值时的灵敏度为88.7%,特异性为93.0%。多因素分析指出血清高TK1、高CD147和高VEGF水平是NSCLC发病的独立危险因素(P<0.05)。结论血清TK1、CD147、VEGF、CYFRA21-1和CEA联合检测在NSCLC临床诊疗意义重大。     

热休克因子在乳腺癌细胞株MCF—7热／化疗前后的表达差异及其影响

目的：探讨乳腺癌细胞株MCF-7经热处理及应用阿霉素前后热休克因子（HSF1）的表达差异及基对热估克蛋白（HSP70）以及P-糖蛋白表达的影响。方法：分别给予乳腺癌细胞株MCF-7非致死温度的热冲击43℃1小时、45℃15分钟以及阿霉素处理，继续培养4小时收获细胞，进行免疫细胞化学染色，观察热/化疗前后热休克因子、热休克蛋白以及P-糖蛋白的表达情况。结果：乳腺癌细胞株MCF-7经上述处理后三组细胞的HSF1、HSP70较处理前蛋白表达有明显增高，43℃组、化疗组P-糖蛋白表达较处理前有明显增高，但45℃组P-糖蛋白的表达在热疗前后无明显变化。结论：经热疗或化疗后的乳腺癌细胞株MCF-7中热休克因子表达增高，并能促进热休克蛋白70及P-糖蛋白的表达；热疗可导致化疗耐药的产生。     

The role of the IL-33/IL-1RL1 axis in mast cell and basophil activation in allergic disorders

Interleukin-33 (IL-33) is a recently discovered cytokine that belongs to the IL-1 superfamily and acts as an important regulator in several allergic disorders. It is considered to function as an alarmin, or danger cytokine, that is released upon structural cell damage. IL-33 activates several immune cells, including Th2 cells, mast cells and basophils, following its interaction with a cell surface heterodimer consisting of an IL-1 receptor-related protein ST2 (IL-1RL1) and IL-1 receptor accessory protein (IL-1RAcP). This activation leads to the production of a variety of Th2-like cytokines that mediate allergic-type immune responses. Thus, IL-33 appears to be a double-edged sword because, in addition to its important contribution to host defence, it exacerbates allergic responses, such as allergic rhinitis and asthma. A major purported mechanism of IL-33 in allergy is the activation of mast cells to produce a variety of pro-inflammatory cytokines and chemokines. In this review, we summarize the current knowledge regarding the genetics and physiology of IL-33 and IL-1RL1 and its association with different allergic diseases by focusing on its effects on mast cells and basophils.     

Systematic review of plasma-membrane ecto-ATP synthase: A new player in health and disease

This review summarizes recent studies on plasma-membrane ecto-ATP synthase from structural and functional standpoints to possible pathophysiological roles. This review discusses significant new contributions and perspectives in the area of ecto-ATP synthase since the topic was last reviewed in 2015. Following an extensive summary of the cell types in which the ecto-ATP synthase is present, its structural and functional mechanism are discussed and physiological and pathological roles of the ecto-ATP synthase are reviewed and evaluated. Attempts to define the possible role of ecto-ATP synthase as possible target for anti-cancer and anti-obesity interventions are discussed.     

Systemic lupus erythematosus and systemic sclerosis: All roads lead to platelets

Systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) are two phenotypically distincts inflammatory systemic diseases. However, SLE and SSc share pathogenic features such as interferon signature, loss of tolerance against self-nuclear antigens and increased tissue damage such as fibrosis. Recently, platelets have emerged as a major actor in immunity including auto-immune diseases. Both SLE and SSc are characterized by strong platelet system activation, which is likely to be both the witness and culprit in their pathogenesis. Platelet activation pathways are multiple and sometimes redundant. They include immune complexes, Toll-like receptors activation, antiphospholipid antibodies and ischemia-reperfusion associated with Raynaud phenomenon. Once activated, platelet promote immune dysregulation by priming interferon production by immune cells, providing CD40L supporting B lymphocyte functions and providing a source of autoantigens. Platelets are actively implicated in SLE and SSc end-organ damage such as cardiovascular and renal disease and in the promotion of tissue fibrosis. Finally, after understanding the main pathogenic implications of platelet activation in both diseases, we discuss potential therapeutics targeting platelets.     

Mechanisms of human smooth muscle cell proliferation and transplant vasculopathy induced by HLA class I antibodies: In vitro and in vivo studies

Vascular smooth muscle cells (SMC) play an important role in the pathophysiology of transplant vasculopathy (TV), a major cause of late death in patients receiving an organ transplant. In this review we describe the proliferative effect in vitro and in vivo of HLA class I antibodies on human SMC. We have developed an experimental model using segments of human mesenteric arteries transplanted in the position of the infrarenal aorta in immunodeficient mice (SCID/beige). Weekly injections of transplanted mice with a monoclonal antibody towards HLA class I provoked typical lesions of TV after 6 weeks in the human graft while transplanted mice receiving an irrelevant antibody did not develop any significant lesion. In vitro, the anti-HLA antibodies were mitogenic to SMC and we showed that they activate a stress-induced signaling pathway implicating matrix metalloproteinases (MMP) and neutral sphingomyelinase 2 (nSMase-2). The proliferative effect of anti-HLA antibodies could be blocked by pharmacological inhibitors or by siRNA. Administration of pharmacological inhibitors diminished the development of TV in grafted mice injected with anti-HLA antibodies demonstrating an important role of the MMP/nSMase-2 pathway in antibody-induced TV. This observation opens new perspectives for the management of TV in clinical settings.     

Effects of lycopene and apigenin on human umbilical vein endothelial cells in vitro under angiogenic stimulation

Angiogenesis is the formation process of new blood vessels from preexisting vessels. Solid tumors need angiogenesis for growth and metastasis. The suppression of tumor growth by inhibition of neoangiogenic processes represents a potential approach to cancer treatment. Lycopene has powerful antioxidant capacities and anticarcinogenic properties. The aim of this study was to investigate the effects of lycopene on angiogenesis in vitro. For this reason, we measured in vitro angiogenesis in human umbilical vein endothelial cells including parameters of cell proliferation, tube formation, cell migration. Lycopene and apigenin were observed to block the endothelial cell proliferation in a dose-dependent manner. In addition, they significantly decreased the capillary-like tube lengths, tube formation and endothelial cell migration. This study provides indications that apigenin and lycopene, which are considered as chemopreventive agents, to be effective in vitro on endothelial cells and angiogenesis.