Tissue factor pathway inhibitor in childhood nephrotic syndrome

It is now recognised that the extrinsic tissue factor pathway is the main trigger to the coagulation system in vivo. Its main inhibitor, tissue factor pathway inhibitor (TFPI), has never been studied in childhood nephrotic syndrome. The aim of the study was to monitor the level of TFPI in childhood nephrotic syndrome. One hundred and thirty-nine nephrotic children were classified into the following groups: group 1 (n=25), in relapse and receiving no treatment; group 2 (n=37), in relapse but receiving steroid treatment; group 3 (n= 45), in early remission and on steroids; group 4 (n=24), in established remission and receiving no steroids; group 5 (n=8), steroid-resistant. The controls (n=84) were healthy and age-matched. There was significant elevation of total TFPI levels in groups 1 and 2 and 3; levels were comparable to those of the healthy controls in group 4. The highest levels of total TFPI were recorded in group 5. Like total TFPI, the levels of the free form of TFPI showed a statistically significant increase in groups 1, 2, 3 and 4, when compared with levels in healthy controls. The highest levels of free TFPI were recorded group 5. We concluded that the elevated levels of both the total and free TFPI in various phases of nephrotic syndrome add another natural anticoagulant mechanism, which will attenuate the hypercoagulability of childhood nephrotic syndrome.     

组织因子途径抑制物2抑制胰腺癌血管生成的研究

目的研究组织因子途径抑制物2（TFPI-2）对胰腺癌血管生成的影响，探讨其抑制胰腺癌生长及侵袭、转移的机制。方法建立裸鼠角膜微囊移植模型，将3组细胞Pane-1TFPI-2、Pane-1-P和Pane-1-V分别接种裸鼠角膜微囊，观察角膜新生血管的形成；再将上述3组细胞接种于裸鼠皮下，观察裸鼠皮下肿瘤生长及转移情况，并采用抗CD34抗体进行血管免疫组织化学染色检测皮下肿瘤的微血管密度（MVD）。Pane-1-TFPI-2组为实验组，Panc-1-P和Pane-1-V组作为对照组。结果实验组角膜新生血管积分比对照组明显减少（P〈0．05），实验组和对照组裸鼠皮下均成瘤，实验组肿瘤体积（438．0±69．8）mm^3，对照组分别为（852．0±102．9）mm^3和（831．0±78．1）mm^3（P〈0．05）；同对照组比较，实验组未见明显远处转移，其肿瘤微血管密度（9．68±1．12），对照组分别为（18．69±2．51）和（20．32±2．08），差异有统计学意义（P〈0．05）。结论组织因子途径抑制物2能抑制肿瘤血管的形成，抑制胰腺癌生长。     

Tissue factor, its inhibitor, and the thrombogenicity of two new synthetic membranes

BACKGROUND: The aim of the study was to compare the effect of new high-flux hemodialysis membranes made from polyacrylonitrile (AN69ST) and polysulfone (Helixone) on the plasma levels of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) playing a key role in hemostasis. Established thrombogenicity markers were also determined. METHODS: In a clinical prospective randomized study, 10 patients were examined using either membrane at the start and at minutes 15, 60, and 240 of hemodialysis. RESULTS: Increases in the plasma TF levels reached significance at the end of hemodialysis with both membranes, with the Helixone also after 15 min. TFPI levels tended to rise significantly from minute 15 onward while not differing from baseline at the end of the procedure. Judging by the increase in thrombin-antithrombin III complexes, both membranes significantly activated coagulation at the end of hemodialysis. Platelet factor 4 levels, released during thrombocyte and endothelial stimulation, were elevated from the start of procedures. There were no significant differences between the AN69ST and the Helixone in any of the assessed markers. CONCLUSIONS: The AN69ST and Helixone membranes do not differ in their effects on TF and TFPI or even in established thrombogenicity markers.     

组织因子途径抑制物-2在肾癌组织中的表达及甲基化

目的 探讨组织因子途径抑制物-2(TFPI-2)对肾癌侵袭性及细胞凋亡的影响,TFPI-2在肾癌组织中的表达与基因启动子甲基化关系.方法 免疫组织化学、蛋白质印迹、荧光定量RT-PCR检测37例肾癌及11例正常肾脏组织TFPI-2基因表达;TUNEL法检测细胞凋亡;荧光定量甲基化特异性PCR检测TFPI-2基因启动子甲基化.结果肾癌与正常肾脏组TFPI-2蛋白质印迹条带吸光度(A)值分别为0.92±0.36、1.61±0.13;2组TFPI-2 mRNA相对表达量为0.0019±0.0011、0.0065±0.0008.随肾癌分期增加,TFPI-2表达下降.肾癌TFPI-2表达1～4级细胞凋亡指数分别为2.41%、3.90%、6.78%、9.57%.随TFPI-2表达增加,肾癌细胞凋亡指数上升.肾癌TFPI-2基因启动子甲基化阳性组与阴性组TFPI-2 mRNA相对表达量分别为0.0015±0.0011、0.0024±0.0009,2组蛋白质印迹条带A值为0.82±0.35、1.04±0.34.基因启动子甲基化阳性组TFPI-2表达低于阴性组.结论 肾癌TFPI-2基因表达与侵袭性呈负相关,促进细胞凋亡;启动子高甲基化是肾癌TFPI-2基因低表达机制之一.     

Tissue factor and its inhibitor in human non-crescentic glomerulonephritis--immunostaining vs plasma and urinary levels.

BACKGROUND: Tissue factor (TF)-the most potent trigger of coagulation and emerging antiapoptotic, proliferative and angiogenic factor, along with its principal inhibitor (tissue factor pathway inhibitor, TFPI) are known to be involved in crescentic glomerulonephritis (GN). We studied the relationship between plasma and urinary levels as well as renal biopsy immunostaining of TF and TFPI antigens with reference to some clinical parameters in human chronic non-crescentic GN. METHODS: We examined plasma and urinary levels of TF and total TFPI (pre-biopsy, ELISA) and the intensity of TF, TFPI 1 and TFPI 2 staining (immunoperoxidase histochemistry) in kidney biopsy specimens from 30 chronic GN patients. RESULTS: Plasma and urinary TF (uTF) were higher in patients than in 18 healthy individuals. In normal kidneys, TF and TFPI 1/2 antigens were undetectable in glomeruli while a distinct staining of both TFPI variants was observed in tubules and interstitial microvessels. In diseased kidneys, TF was strongly expressed in glomeruli but was undetectable in tubules. In contrast, staining for TFPI 1/2 was observed in glomeruli and tubules. Neither plasma nor urinary levels of the markers correlated with the intensity of TF and TFPI 1/2 staining in biopsy specimens. uTF was significantly associated with creatinine clearance (R = 0.489, P = 0.006) and urinary TFPI (R = 0.554, P = 0.014), and tended to be lower in proliferative vs non-proliferative GN [83 (0-617) vs 281 (10-805) pg/ml; P = 0.06]. CONCLUSION: The intrarenal TF/TFPI system is profoundly disturbed in chronic GN. Plasma and urinary concentrations of TF and TFPI probably do not reflect genuine activity of the disease, likely due to a confounding effect of kidney insufficiency. uTF measurement seems to be helpful in initial identification of proliferative GN, yet further studies are required to validate its use as a marker of glomerular injury in chronic GN.     

Dynamically monitoring tissue factor and tissue factor pathway inhibitor following secondary brain injury

Objective: To study the altering rule of coagulation function at molecular level in patients with secondary brain injury (SBI). Methods: Tissue factor (TF) and tissue factor pathway inhibitor (TFPI) were studied in 32 patients 1, 2,3 and 7 days after craniocerebral injury. Repeated cranial CT scans and platelet counts were made simultaneously.Same measurements were done in 30 normal adults except CT scan. Results: No obvious difference was found in age, sex and platelet count between the injured and the normal groups. TFPI/TF decreased markedly in the first week after injury in patients with SBI, but only decreased on the 7th day in the patients without obvious SBI. For the patients who developed delayed intracranial hematoma (DIH) or hematoma enlargement, TF rose only 1 and 2 days after injury, but TFPI had a tendency to rise again after a fall on the 3rd day. For those patients who developed no DIH, TF rose all the time within the 1st week. Conclusions: Decrease of TFPI/TF for a long time,especially within 3 days after injury, may be one of the most important reasons for SBI. High expression of TF for a relative short time and increase of TFPI after a fall within 3 days may be one of the important reasons for DIH or hematoma enlargement.     

中国汉族深静脉血栓形成患者与组织因子途径抑制物基因多态性的关系

目的探讨组织因子途径抑制物(TFPI)基因变异与中国汉族深静脉血栓形成(DVT)患者的相关性。方法采用聚合酶链反应单链构象多态性分析及DNA测序,对62例DVT患者和50例正常汉族人进行TFPI基因序列分析。结果8号内含子中发现一个C/T多态位点,等位基因频率为86.6%和13.4%,病例组与对照组比较无统计学差异;5号内含子41位发现一个单碱基“G”插入,病例组中有4例(6.5%)与对照组比较差异无统计学意义(P>0.05)。结论TFPI基因可能不是中国汉族人DVT的主易感基因;5号内含子中单核苷酸“G”插入有深入研究价值。     

凝血因子Xa直接抑制剂的研究现状及应用

抗凝药物在血栓性疾病的预防和治疗中具有重要地位。抗凝治疗是防治静脉血栓栓塞症（VTE）、心房颤动（AF）和急性冠状动脉综合征（ACS）的重点,由于传统药物的局限性,凝血因子Xa抑制剂成为了研究的热点药物并逐步走向临床。本文对凝血因子Xa直接抑制剂的研究现状进行回顾,并对其临床应用及临床试验研究结果进行综述。     

组织因子途径抑制物2在肿瘤发生转移和侵袭中的作用

细胞外基质(extracellularmatrix,ECM)降解是肿瘤发生浸润转移过程中的关键一步,肿瘤细胞分泌的多种丝氨酸蛋白酶包括基质金属蛋白酶(matrixmetallopoteinases,MMPs)参与细胞外ECM降解。组织因子途径抑制物2(tissuefactorpathwayinhibitor-2,TFPI-2)是一种具有32000丝氨酸蛋白酶抑制物,可抑制包括纤溶酶、胰蛋白酶、MMPs在内的多种蛋白酶。越来越多的研究表明,TFPI-2通过降解MMPs等抑制肿瘤转移、降低肿瘤的侵袭转移能力。TFPI-2在抑制肿瘤细胞迁徙及浸润转移过程中发挥重要作用,可作为一种潜在的基因治疗靶点。     

TFPI-2对胰腺癌细胞体内和体外侵袭能力的影响

目的 研究组织因子途径抑制物-2(TFPI-2)基因对人胰腺癌细胞系Panc-1细胞侵袭能力的影响.方法 通过脂质体法将TFPI-2基因转染到Panc-1细胞, 用RT-PCR和Western blot分别检测转染前、后TFPI-2 mRNA和相应蛋白的表达.Boyden小室法测定Panc-1-TFPI-2、Panc-1-V和Panc-1-P 3组细胞的体外侵袭能力的变化,同时将3组细胞分别接种裸鼠,观察其体内肿瘤生长及转移情况.结果 转染成功的Panc-1细胞有TFPI-2 mRNA及相应蛋白的表达; Panc-1-TFPI-2组、Panc-1-V组和Panc-1-P组细胞穿膜细胞数分别为(24.4±3.5)个、(61.3±4.1)个和(60.2±3.9)个,前者明显少于后两者(P＜0.05),提示Panc-1-TFPI-2组细胞的体外侵袭能力下降.3组细胞分别接种裸鼠,Panc-1-TFPI-2组肿瘤体积为(438.0±69.8) mm3,与Panc-1-V组的(852.0±102.9) mm3和Panc-1-P组的(831.0±78.1) mm3相比明显缩小,差异有统计学意义(P＜0.05); 镜下见Panc-1-V组和Panc-1-P组肿瘤组织浸润到肌层,有肝、肺转移灶,而Panc-1-TFPI-2组未见肌层浸润及肝、肺转移.结论 TFPI-2基因表达可抑制胰腺癌细胞系Panc-1的侵袭转移能力,为胰腺癌的基因治疗提供了实验依据.     

Altered Plasma Antigen Levels of Tissue Factor Pathway Inhibitor During Open-Heart Surgery

r = 0.96, P < 0.0001) which increased significantly after heparin injection (P < 0.0001), and increased further during the bypass period (P < 0.005). The increased free TFPI antigen level during CPB correlated with the duration of bypass (r = 0.65, P = 0.02). When heparin was neutralized by protamine, the free TFPI antigen level decreased immediately, but remained higher than the preoperative level (P < 0.005). These results suggest that plasma TFPI antigen levels increase during CPB. (Received for publication on Dec. 14, 1998; accepted on July 13, 1999)     

组织因子途径抑制物-2影响人前列腺癌细胞侵袭能力的实验研究

目的:探讨人组织因子途径抑制物2(TFPI-2)对基因转染入人前列腺癌细胞株PC3M体外和体内侵袭转移能力的影响.方法:通过Boyden小室体外侵袭转移模型,评价转染成功的PC3M-TFPI-2细胞和未转染的PC3M迁徙和侵袭能力的强弱;并且分别进行裸鼠接种实验,观察两组间其形成的局部新生物相对体积、组织浸润程度,以及有无远处转移的差异.结果:转染成功的PC3M-TFPI-2细胞中,证实有TFPI-2mRNA和相应蛋白质的表达,转染成功的PC3M-TFPI-2细胞较未转染的PC3M体外侵袭能力明显下降,接种PC3M-TFPI-2组细胞的裸鼠较接种未转染的PC3M的裸鼠形成的新生物肌层浸润明显减少.结论:人TFPI-2基因真核表达载体成功转染人PC3M并可以在PC3M中得到表达,TFPI-2的表达可以抑制PC3M的体外和体内的侵袭能力.     

MMP-1、TIMP-1及PDGF在病理性瘢痕中的表达

目的研究基质金属蛋白酶-1(MMP-1)、金属蛋白酶组织抑制剂-1(TIMP-1)、血小板源性生长因子(PDGF)在病理性瘢痕中的表达及意义。方法对58例病理性瘢痕手术切除标本采用免疫组化方法。结果MMP-1、TIMP-1、PDGF在病理性瘢痕中呈阳性表达(62.07%,63.71%,55.17%),在正常皮肤与普通瘢痕中几乎无阳性表达,病理性瘢痕组与两对照组差异均有显著意义(P<0.05)。病理性瘢痕中TIMP-1与PDGF呈正相关(r=0.331,P<0.05)。结论TIMP-1、PDGF促进病理性瘢痕形成;在病理性瘢痕形成过程中PDGF可促进TIMP-1的表达;病理性瘢痕的形成与MMP-1、TIMP-1、PDGF相互作用失衡有关。     

碱性成纤维细胞生长因子对大鼠烫伤创面中基质金属蛋白酶及其抑制剂的影响

目的　观察大鼠深Ⅱ度烫伤创面愈合过程中基质金属蛋白酶 (MMP) 2、MMP 7与金属蛋白酶组织抑制因子 2 (TIMP 2 )的变化以及创面外用碱性成纤维细胞生长因子 (bFGF)对其的影响。　方法　制作 30 %TBSA深Ⅱ度烫伤大鼠模型 ,分为单纯烫伤组和bFGF治疗组。于伤后 3、6h和 1、3、7、14d取创面皮肤标本 ,检测再上皮化率及胶原含量 ,并采用免疫组织化学染色法检测创面组织真皮内成纤维细胞MMP 2、MMP 7和TIMP 2的表达变化。另取 6只正常大鼠作为假烫组 ,分别检测上述各项指标。　结果　 (1)伤后 3～ 14d ,bFGF治疗组再上皮化率高于单纯烫伤组。 (2 )伤后 3h～ 3d ,bFGF治疗组和单纯烫伤组胶原含量持续下降 ,7～ 14d开始回升 ,但仍低于假烫组 (P<0 .0 5 )。 (3)伤后 1d ,单纯烫伤组MMP 2、MMP 7和TIMP 2表达增多 ,7d时达到高峰 ,持续到 14d。(4 )伤后 3～ 6h,bFGF治疗组MMP 2、MMP 7的表达与单纯烫伤组相似 ;伤后 1～ 14d ,3者的阳性表达强于单纯烫伤组。　结论　烫伤大鼠创面中细胞外基质的活动会影响皮肤的胶原沉积 ;MMP 2、MMP 7、TIMP 2的表达变化是组织修复的重要步骤 ,与bFGF加速创面愈合的过程密切相关。     

组织因子表达与直肠癌肝转移的关系及其调控

目的探讨组织因子(TF)在原发直肠癌中的表达意义,初步分析血管内皮生长因子(VEGF)对人直肠癌细胞(HT-29)TF表达的调控作用。方法(1)应用免疫组织化学染色法研究40例原发直肠癌组织、6例直肠良性腺瘤组织TF表达特点,分析其与转移和预后的关系;(2)以VEGF(25ng/ml)孵育HT-29细胞,分别应用免疫印记法、细胞免疫组织化学染色法和促凝血活性分析法检测不同时间点HT-29细胞TF的表达水平及活性。结果(1)40例原发直肠癌中20例(50%)TF表达阳性,正常黏膜和直肠良性腺瘤无TF表达,多因素回归分析表明,TF是影响原发直肠癌预后的因素之一(P=0.024);(2)直肠癌同时性和异时性肝转移组TF的表达均与肝转移显著相关(P=0.002和0.001);(3)Logistic回归分析表明,TF为直肠癌肝转移的影响因素(P=0.003);(4)静止状态下HT-29细胞存在TF基础表达,VEGF(25ng/ml)可诱导HT-29细胞TF表达增强,6h达高峰,刺激效应可维持24～32h。结论(1)TF可能参与原发直肠癌肝转移的生物学过程,其阳性表达可能作为高危患者术后肝转移的预测指标应用于临床;(2)TF是影响原发直肠癌预后的因素之一;(3)VEGF可能参与直肠癌细胞组织因子表达的上调过程。     

MMP-1、TIMP-1及PDGF在病理性瘢痕中的表达

目的研究基质金属蛋白酶-1（MMP-1）、金属蛋白酶组织抑制剂-1（TIMP-1）、血小板源性生长因子（PDGF）在病理性瘢痕中的表达及意义。方法对58例病理性瘢痕手术切除标本采用免疫组化方法。结果MMP-1、TIMP-1、PDGF在病理性瘢痕中呈阳性表达（62．07％，63．71％，55．17％），在正常皮肤与普通瘢痕中几乎无阳性表达，病理性瘢痕组与两对照组差异均有显著意义（P〈0．05）。病理性瘢痕中TIMP-1与PDGF呈正相关（r=0．331，P〈0．05）。结论TIMP-1、PDGF促进病理性瘢痕形成；在病理性瘢痕形成过程中PDGF可促进TIMP-1的表达；病理性瘢痕的形成与MMP-1、TIMP-1、PDGF相互作用失衡有关。     

Tissue factor, tissue factor pathway inhibitor and vascular endothelial growth factor-A in carotid atherosclerotic plaques.

OBJECTIVE: To determine the concentration of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and vascular endothelial growth factor A (VEGF-A) in carotid plaques. MATERIALS AND METHODS: Thirty-eight consecutive patients (20 symptomatic, 18 asymptomatic) undergoing carotid endarterectomy were enrolled into the current study. The concentration of TF, TFPI and VEGF-A in carotid plaque homogenates and blood plasma was measured using enzyme immunoassay. RESULTS: The concentration of TF in carotid plaque homogenates was 60 fold higher than in blood plasma. There were no statistically significant differences between the concentration of TF, TFPI and VEGF-A in symptomatic and asymptomatic plaques. Carotid plaques of diabetic patients contained an increased level of TF and VEGF-A ( p = 0.002, p = 0.005). The plaque concentration of VEGF-A was elevated among older patients ( p = 0.02). Carotid plaques of non-smokers contained an increased level of TFPI ( p = 0.03). The concentration of TF, TFPI and VEGF-A in carotid plaques correlated positively with plasma level of these factors ( R = 0.86; p < 0.0001; R = 0.91; p < 0.0001; R = 0.80; p = 0.001, respectively). A highly positive correlation between concentration of VEGF-A and TF, TFPI in carotid plaques was also observed ( R = 0.75; p < 0.001; R = 0.62; p < 0.001, respectively). CONCLUSIONS: TF, TFPI and VEGF-A concentrations do not differ in atheroma removed from symptomatic and asymptomatic patients but are higher in diabetic patients. There is a highly positive correlation between the level of VEGF-A and TF, TFPI in carotid plaques.     

力生长因子的表达及其在组织修复与再生中的作用

目的对力生长因子(mechano-growth factor,MGF)的表达以及在组织修复与再生中的作用进行综述。方法广泛查阅近年来国内外有关MGF表达和参与组织修复与再生的文献,对其研究进展进行综述。结果 MGF在多种组织/细胞中表达,具有力敏感性,并能激活卫星细胞,促进成肌细胞增殖,在治疗肌肉缺损、预防心肌损伤和修复受损神经等方面起重要作用。结论 MGF在多种组织中的修复作用已明确,但其具体作用机制尚未明确,有待进一步深入研究。