冠状动脉TF/TFPI与急性冠脉综合征关系的研究

目的：研究急性冠脉综合症患者冠状循环局部血液TF/TFPI改变。方法：入选40名冠状动脉造影患者,其中不稳定心绞痛（Unstable pectoris,UP）20例,稳定性心绞痛（Stable pectoris,SP）和正常对照组各10例。所有受试者在冠状动脉造影时,同时经导管抽取主动脉根部、冠状静脉窦和股静脉血样,采用ELISA法测定血浆TF、TFPI水平,计算TF/TFPI比值。结果：三组受检者基本情况均相似,UA组和SA组冠心病危险因素无统计学差异。股静脉血中TF、TFPI浓度及TF/TFPI比值比较：UA组和SA组血TF水平显著高于对照组,UA组血TF水平显著高于SA组。UA组血TFPI水平低于SA组及对照组;SA组血TFPI水平略低于对照组,但未达到统计学意义;UA组血TF/TFPI比值显著高于对照组和SA组,SA组与对照组比较差异无统计学意义。不同部位血TF、TFPI水平及TF/TFPI比值比较：UA组冠状静脉窦血中TF水平较主动脉根部显著升高,CS-AO差值显著高于FV-AO差值;UA组冠状静脉窦血TFPI水平较主动脉根部有升高趋势,但差异无统计学意义,而TF/TFPI比值较主动脉根部升高,差异有统计学意义,CS-AO差值显著高于FV-AO差值。血TF、TFPI水平及TF/TFPI比值在股静脉与主动脉根部之间差异无统计学意义（P〉0.05）。SA组和对照组血TF、TFPI水平及TF/TFPI比值三部位间差异无统计学意义。结论：冠脉循环TF/TFPI比值能较敏感地反映ACS的情况。     

胞内钙与蛋白激酶C对人脐静脉内皮细胞组织因子途径抑制物及组织因子表达的影响

为探讨细胞内钙及蛋白激酶C在血管内皮细胞释放组织因子（TF）,组织因子途径抑制物（TFPI）中的作用,本文采用钙离子载入剂A23187与蛋白激酶C激动剂佛波脂（PMA）刺激原代培养人脐静脉内皮细胞（HUVEC）,观察各实验组内皮细胞冻融液中TF活性及条件培养液中TFPI活性的变化,以一期凝固法测TF活性,二期生色法测定TFPI活性。结果显示：A23187,PMA及A23187＋PMA组TF活性较对照明显升高（P＜0.001）,三组中A23187和A23187＋PMA组活性明显低于PMA组（P＜0．05）,但前两组间无显著性差异（P＞0．05）;A23187组TFPI活性与对照无明显差别,但PMA与A23187＋PMA组TFPI活性较对照及A23187组明显升高（P＜0.01）。结果提示：细胞内钙升高及PKC激活促进血管内皮细胞TF的合成,以PKC为强;PKC促进血管内皮细胞释放TFPI,而钙对此无明显作用。     

急性白血病患者血浆VEGF、TF水平测定及其临床意义

目的:研究血清中血管内皮生长因子(VEGF)和组织因子(TF)的表达及其与急性白血病发生和发展的关系.方法:收集70例急性白血病患者(AL),其中初发未治者31例,完全缓解者14例,未缓解者7例,复发患者18例,另外选择20名健康体检者作为对照组,应用酶联免疫吸附试验(EL ISA)法测定各组血清中VEGF及TF的含量,并进行统计学分析.结果:AL初发未治组、未缓解组、复发组患者血清VEGF和TF的含量均明显高于正常对照组和缓解组(P<0.01);而缓解组患者血清VEGF和TF的含量与正常对照组比较其差异无统计学意义(P>0.05);TF含量与VEGF含量在AL中呈显著正相关(r=0.675,P<0.01).结论:VEGF和TF与AL的发生、发展密切相关.动态检测急性白血病患者的血清VEGF及TF水平对观察病情、预测复发、指导治疗均有十分重要的临床价值.     

两种剂量阿托伐他汀治疗老年急性冠脉综合征的疗效及影响炎症、凝血因子的研究

目的:探讨两种剂量阿托伐他汀治疗老年急性冠脉综合征的疗效及对炎症、凝血因子的影响。方法:选择我院2010年1月~2012年12月收治的120例老年急性冠脉综合征患者作为观察对象,根据住院号随机分为观察组和对照组,每组均60例,两组患者均采用阿托伐他汀治疗,对照组予以10 mg/d,观察组予以20 mg/d,比较两组的临床效果以及炎症因子、凝血因子的变化。结果:治疗后,两组血脂达标率均显著提高,观察组治疗后1个月的血脂达标率为33.3%,治疗后3个月的血脂达标率为46.7%,均显著高于对照组的15.0%、23.3%,差异均具有统计学意义(均P0.05);治疗后1个月和3个月所有患者血浆CRP和TF均显著下降,观察组治疗后1个月和3个月CRP和TF水平均显著低于对照组,差异均具有显著性(均P0.05);观察组治疗后3个月TFPI水平显著高于对照组,差异具有统计学意义(P0.05)。结论:大剂量阿托伐他汀治疗老年急性冠脉综合征患者临床疗效优于小剂量,对炎症因子和凝血因子的影响有利于预后的改善,值得临床进一步推广应用。     

血UA水平与急性脑梗死患者颈动脉斑块及梗死分型的关系

目的:探讨血尿酸(UA)水平与急性脑梗死(ACI)患者颈动脉斑块及梗死分型的关系。方法:选择2013年11月~2016年5月本院收治的ACI患者200例,依据牛津郡社区卒中研究分型(OCSP)标准将患者分为完全前循环梗死(TACI)组,部分前循环梗死(PACI)组、后循环梗死(POCI)组和腔隙性梗死(LACI)组;依据斑块性质将所有患者分为无斑块组、稳定斑块组和不稳定斑块组,分析不同梗死分型和斑块性质患者血UA水平差异。结果:TACI组与PACI组、POCI组与LACI组组间UA水平比较差异无统计学意义(P0.05),而TACI组和PACI组均高于POCI组和LACI组,差异有统计学意义(P0.05);UA水平与OCSP分型存在相关性(r=0.237,P=0.001);无斑块组UA水平与稳定斑块组的差异无统计学意义(P0.05);不稳定斑块组UA水平显著高于无斑块组和稳定斑块组,差异有统计学意义(P0.05),UA水平与不同斑块性质分型存在相关性(r=0.356,P=0.000);ACI患者总体OCSP分型和斑块性质存在相关性(r=0.334,P=0.000)。结论:血UA水平与ACI患者颈动脉斑块及梗死分型存在相关性,应重视ACI患者UA水平的检测,以预防脑梗死。     

左房内径、血清尿酸水平与老年心房颤动的相关性分析

目的:探讨左房内径(LAD)、血清尿酸(UA)水平与老年心房颤动的相关性。方法:选择2013年1月至2016年7月在我院住院的60岁以上的非瓣膜性房颤患者,共166例,其中持续性房颤组85例,阵发性房颤组81例,选择同期无房颤的高血压、冠心病老年患者83例作对照组。通过心脏彩超检查检测LAD、左心室舒张末期内径(LVDD)、左心室收缩末期内径(LVDS)、左心室射血分数(LVEF),以≥40 mm为左房内径增大。并采用生化分析检测患者血清UA水平。结果:(1)持续性房颤组LAD、LVEF、左心房增大发生率均显著高于阵发性房颤组和对照组,而阵发性房颤组以上指标均明显高于对照组,差异具有统计学意义(P0.05)。(2)持续性房颤组和阵发性房颤组患者血清UA水平均显著高于对照组,但持续性房颤组和阵发性房颤组之间血清UA水平比较差异无统计学意义(P0.05)。结论:左心房内径大小、血尿酸水平与老年患者心房颤动的发生密切相关。     

不同浓度的尿酸对软骨细胞活性及炎症因子表达的影响

目的：检测软骨细胞在不同浓度尿酸（uric acid, UA）环境下的活性及其白介素-1beta（IL-1beta）和肿瘤坏死因子-alpha（TNF-alpha）的表 达水平的变化,探讨不同浓度的尿酸对软骨的影响。方法：分别用0、4、8、10、16、32 mg/dL的尿酸溶液培养大鼠关节软骨细胞, MTT 比色法检测软骨细胞活力,ELISA 方法检测细胞培养液中IL-1beta、TNF-alpha的浓度。结果：4 mg/dL UA 组软骨细胞活性低于对 照组（0 mg/dL),差异无统计学意义（P＞0.05);8、10、16、32 mg/dL UA 组软骨细胞活性均低于对照组,差异有统计学意义（P＜ 0.05),且其活性随着尿酸浓度的升高而降低。4、8、10 mg/dL UA 组IL-1beta、TNF-alpha的水平均高于对照组,差异有统计学意义（P＜ 0.05),其水平随尿酸浓度的升高而增加。10、16、32 mg/dL UA 组IL-1beta、TNF-alpha的水平亦均高于对照组,差异有统计学意义（P＜ 0.05),但其水平随着尿酸浓度的升高而降低。结论：尿酸可以抑制软骨细胞的活性;同时,在4-10 mg/dL浓度范围内明显促进软骨 细胞IL-1beta、TNF-alpha的表达,但高于10 mg/dL后,随着细胞活性的严重降低其促进作用逐渐减弱。这可能是尿酸参与软骨破坏及 骨关节炎发生的重要因素。     

血清尿酸水平对急性脑梗死患者颈动脉粥样硬化斑块的影响

目的:探讨血清尿酸(UA)水平对急性脑梗死患者颈动脉粥样硬化斑块的影响。方法:回顾性分析2011年6月至2016年1月我院收治的251例急性脑梗死患者的临床资料,根据有无颈动脉粥样硬化斑块分为伴颈动脉粥样硬化斑块组(观察组)176例和无颈动脉粥样硬化斑块组(对照组)75例,观察组根据颈动脉粥样硬化程度分为斑块形成组(113例)、内中膜增厚组(63例),根据颈动脉斑块稳定程度分为不稳定组(106例)、稳定组(70例),比较各组血清UA水平,根据UA水平不同分为高UA组(134例)和正常UA组(117例),进行颈动脉斑块发生情况比较。结果:观察组的血清UA水平显著高于对照组,差异有统计学意义(P0.05)。(1)斑块形成组和内中膜增厚组血清UA水平显著高于对照组(P0.05),而斑块形成组和内中膜增厚组血清UA水平比较,差异无统计学意义(P0.05);(2)不稳定组血清UA水平显著高于对照组和稳定组(P0.05),而稳定组和对照组血清UA水平比较,差异无统计学意义(P0.05);(3)正常UA组和高UA组颈动脉斑块的发生情况比较,差异无统计学意义(P0.05)。结论:血清UA水平可以作为表征急性脑梗死患者伴随出现颈动脉粥样硬化斑块的生物学指标之一,此外,血清UA的水平在颈动脉粥样硬化斑块形成者和不稳定者表达更高,但血清UA水平与颈动脉斑块形成无明显联系。     

过敏性紫癜患儿血浆白细胞介素9水平变化及意义探讨

目的:探讨儿童过敏性紫癜急性期血浆白细胞介素9(IL-9)水平变化特点及其临床意义。方法:观察组为79例过敏性紫癜(HSP)患儿,分为无肾损害组和有肾损害两组;对照组为28例门诊健康体检儿童。ELISA测定各组儿童血浆IL-9及白细胞介素4(IL-4)水平。结果:HSP患儿血浆IL-9水平为(28.32±6.97)pg/ml,显著高于对照组(23.92±5.91)pg/ml,差异有统计学意义(t=2.98,p0.05);HSP患儿血浆IL-4水平为(93.86±25.35)pg/ml,显著高于对照组(77.75±15.90)pg/ml,差异有统计学意义(t=3.01,p0.05);紫癜有肾损害组及紫癜无肾损害组血清IL-4及IL-9水平较对照组明显升高,差异有统计学意义,p0.05。紫癜有肾损害组血清IL-9水平较紫癜无肾损害组明显升高,差异有统计学意义,p0.05。紫癜有肾损害组血清IL-4水平较紫癜无肾损害组升高不明显,差异无统计学意义,p0.05。直线相关分析结果显示,HSP患儿血浆IL-9水平变化与IL-4水平呈显著正相关(r=0.298,p0.05)。结论:IL-9水平的显著增高在HSP特别是紫癜性肾炎发病机制中有重要作用。     

金黄色葡萄球菌LukF-PV基因的原核表达及血清特异性抗体的检测

目的:在大肠杆菌中表达金黄色葡萄球菌(Staphylococcus aureus,SA)LukF-PV基因,纯化重组蛋白并以其为抗原检测儿童SA感染者血清特异性IgG抗体,分析不同部位SA感染患者血清LukF-PV抗体水平。方法:将Luk F-PV克隆至pET-28a(+)载体,IPTG诱导重组蛋白表达、His柱纯化重组蛋白后,用间接ELISA检测儿童SA感染者与健康对照者血清特异性IgG抗体水平。结果:成功表达纯化LukF-PV蛋白,儿童SA感染组与健康对照组血清特异性抗体均值分别为(0.309±0.063)、(0.505±0.261),具有统计学意义(P0.05)。不同部位来源标本的感染者中血液组和扁桃体组抗体OD均值分别为(0.634±0.225)、(0.481±0.264)与健康对照组有显著差异(P0.05),其余各组与健康对照组无显著差异(P0.05)。抗体阳性率统计分析中血液组分别与脑脊液组、扁桃体组、咽拭子组、痰液组有统计学意义(P0.05),其余各组间均无显著差异(P0.05)。结论:LukF-PV成功表达纯化,,儿童SA感染组LukF-PV特异性IgG抗体显著高于健康对照组,其中来源于血液和扁桃体部位的SA感染者抗体水平与健康者有显著差异,尤以血液感染者最显著。     

Tumour-expressed tissue factor inhibits cellular cytotoxicity

Aims: The association between tissue factor (TF) expression and increased rate of tumour metastasis is well established. In this study, we have examined the hypothesis that the expression of TF by disseminated tumour cells confers protection against immune recognition and cytotoxicity. Materials and methods: A hybrid EGFP-TF protein was expressed in HT29 colon carcinoma and K562 lymphoblast cell lines. To assess the cytotoxic activity against tumour cells over-expressing TF, a novel method was used, based on the direct measurement of fluorescently labelled HT29 or K562 target cells. Results: Upon challenge with peripheral blood mononuclear cells (PBMC), tumour cells expressing TF partially evaded cellular cytotoxicity (Δ=15–40% reduction in cytotoxicity). Moreover, the influence of TF was not primarily dependent on its procoagulant function, although the inclusion of 20% (v/v) plasma did lower the rate of cytotoxicity against untransfected cells. However, expression of a truncated form of TF, devoid of the cytoplasmic domain, did not mediate any degree of inhibition of cytotoxicity, suggesting that the protective function of TF is principally due to this domain. Conclusions: We conclude that TF can promote immune evasion in tumour cells expressing this protein leading to increased survival and therefore metastatic rate in such cells.     

Synergistic induction of endothelial tissue factor by tumor necrosis factor and vascular endothelial growth factor: functional analysis of the tumor necrosis factor receptors

Tissue factor expression on the surface of endothelial cells can be induced by tumor necrosis factor (TNF) and vascular endothelial growth factor (VEGF) in a synergistic manner. We have investigated the role of the two different TNF receptors for this synergy. Firstly, stimulation of the 60 kDa TNF receptor (TNFR60) by a mutant of TNF specific for TNFR60 induced responses comparable to wild-type TNF. In contrast, stimulation of TNFR80 by a TNFR80-specific TNF mutein did not result in enhancement of tissue factor expression even in the presence of a suboptimal TNFR60 triggering. Secondly, we tested neutralizing TNF receptor antibodies for inhibition of tissue factor synthesis induced by VEGF and TNF. A TNFR60-specific antibody inhibited tissue factor production over a broad range of TNF concentrations, indicating an essential role of TNFR60 in the TNF/VEGF synergy. In contrast, blocking of TNF binding to TNFR80 strongly inhibited TNF-induced tissue factor expression at low, but less pronounced at high, TNF concentrations. In conclusion, these data are in agreement with a model in which TNFR80 participates in the synergy between VEGF and low concentrations of soluble TNF by passing the ligand to the signalling TNFR60.     

Antisense oligonucleotide for tissue factor inhibits hepatic ischemic reperfusion injury

Tissue factor (TF) is an initiation factor for blood coagulation and its expression is induced on endothelial cells during inflammatory or immune responses. We designed an antisense oligodeoxynucleotide (AS-1/TF) for rat TF and studied its effect on hepatic ischemic reperfusion injury. AS-1/TF was delivered intravenously to Lewis rats. After 10 h, hepatic artery and portal vein were partially clamped. Livers were reperfused after 180 min and harvested. TF expression was studied using immunohistochemical staining. One of 10 rats survived in a 5-day survival rate and TF was strongly stained on endothelial cells in non-treatment group. However, by treatment with AS-1/TF, six of seven survived and TF staining was significantly reduced. Furthermore, we observed that fluorescein-labeled AS-1/TF was absorbed into endothelial cells. These results suggest that AS-1/TF can strongly suppress the expression of TF and thereby inhibit ischemic reperfusion injury to the rat liver.     

Cellular localization of tissue factor in human breast cancer cell lines

Expression of tissue factor (TF), the cellular receptor of clotting factor VII/VIIa, is a feature of certain malignant tumours. The TF gene has been classified as an immediate early gene responsive to serum and cytokines. Thus, the regulation of TF gene expression seems to play a role in cell growth. Recently, we have shown that constitutive TF expression in MCF-7 breast cancer cells is modulated by such growth factors as EGF, TGFα, and IL-1. The present study deals with the immunocytochemically detectable cellular distribution of TF in human breast cancer cell lines MCF-7 and MaTu stimulated by EGF and TGFα. In MCF-7 cells growing logarithmically, stimulation led to a significant increase of TF mRNA after 2 h (in situ hybridization, Northern blot) and to maximum TF expression after 6 h (immunohistochemistry). When decorated by monoclonal antibodies, TF protein showed a pronounced localization at ruffled membrane areas, cell edges, and processes of spreading cells after 6 and 20 h. In more flattened cells TF was concentrated in peripheric lamellae and microspikes communicating with neighbouring cells. After epithelial colony pattern had established, TF was predominantly accumulated at the intercellular boundaries. The vary same distribution patterns as seen in MCF-7 cells were true for the stimulated MaTu cell line. The dynamics and cellular distribution patterns of stimulated TF expression support the hypothesis that TF could be of importance for morphogenic events associated with the growth and differentiation of breast cancer cells in culture.     

Proteolytic activation of tissue plasminogen activator by plasma and tissue enzymes

Tissue kallikrein and factor Xa were found to activate tissue plasminogen activator (t-PA) at a rate comparable with that of plasmin. During the activation reaction, the single-chain molecule was converted into a two-chain form. A slight t-PA activating activity was also found in plasma kallikrein. Other activated coagulation factors, factor XIIa, factor XIa, factor IXa, factor VIIa, thrombin and activated protein C had no effect on t-PA activation. t-PA was also activated by a tissue kallikrein-like enzyme that was isolated from the culture medium of melanoma cells. These results indicate that tissue kallikrein and factor Xa may participate in the extrinsic pathway of human fibrinolysis.     

Thrombophilia in mice expressing a tissue factor variant lacking its transmembrane and cytosolic domain

Mice with a targeted truncation in the gene encoding tissue factor of blood coagulation (TF) to eliminate the cytosolic domain and carrying a neo(R) cassette in intron 5 unexpectedly displayed severe spontaneous thrombosis in various vascular beds. Thrombosis was observed in heterozygous TF(+/neo) mice, causing death of over 50% of adults within 36 weeks of birth, and fulminantly exacerbating in pregnant females. Homozygous TF(neo/neo) mice were more severely affected and died within 7 weeks after birth. These TF(neo) mice primarily synthesized a mutant mRNA aberrantly spliced from exon 5 to neo(R), encoding an apparently non-vesicle-binding soluble TF lacking both the transmembrane and cytosolic domain, but still capable of blood coagulation induction. This severe thrombotic phenotype associated with the presence of a non-anchored soluble TF variant underscores the recently recognized significance of circulating TF for thrombus formation and development.     

血肌酐与非ST段抬高急性冠脉综合征冠脉病变程度及其预后

目的:研究血清肌酐(serum creatinine,SCr)与非ST段抬高急性冠脉综合征(non-ST-elevation acute coronary syndrome,NSTE-ACS)患者冠脉病变程度及其预后的关系。方法:对293例患者进行回顾性分析,依据冠脉造影结果分为NSTE-ACS组和非冠心病组。根据Gensini积分系统,评价NSTE-ACS患者冠脉病变程度,并将患者分为轻度病变组、中度病变组和重度病变组。检测患者SCr水平,应用SPSS16.0分析SCr与NSTE-ACS患者冠脉病变程度及其预后关系。结果:(1)与非冠心病组相比,NSTE-ACS患者SCr较高(P0.05);其中,重度冠脉病变NSTE-ACS患者SCr水平尤高(P0.001)。(2)SCr是NSTE-ACS冠脉病变的危险因子;SCr与NSTE-ACS冠脉病变程度呈正相关(r=0.263,P0.000);SCr与NSTE-ACS主要心血管不良事件(Major Adverse Cardiovascular Events,MACE)呈正相关(r=0.183,P0.01)。结论:SCr是NSTE-ACS患者冠脉病变的独立危险因子,且与NSTE-ACS患者预后相关。SCr对于NSTE-ACS的诊疗有潜在临床价值。     

人脐静脉血管平滑肌促内皮细胞组织因子表达的体外实验研究

目的研究血管平滑肌细胞对血管内皮细胞组织因子表达的影响并探讨其临床意义.方法用贴块法培养人脐静脉平滑肌细胞;酶消化法培养人脐静脉内皮细胞;用培养平滑肌细胞条件培养液(SMC-CM)刺激培养的内皮细胞,一步凝固法检测内皮细胞组织因子的活性;Northern blot检测内皮细胞组织因子的mRNA表达;并用酶联免疫吸附试验检测SMC-CM中IL-1α、IL-1β、TNF-α和VEGF的含量.结果 SMC-CM使内皮细胞组织因子活性呈剂量依赖性增强,作用6h增至最高,最高增强约38倍;SMC-CM使内皮细胞组织因子mRNA表达显著增强;SMC-CM中的组织因子诱导剂不耐热,且并非IL-1α、IL-1β、TNF-α和VEGF等已知的组织因子诱导剂.结论血管平滑肌细胞能促进血管内皮细胞组织因子的表达,提示体内增生的平滑肌细胞,如动脉再狭窄新内膜中的平滑肌细胞可能诱导局部血管内皮细胞活化及表达组织因子,在局部血栓形成中起一定作用.