Tissue factor expression in colorectal adenocarcinoma: association with angiogenesis and clinical and pathological aspects

Introduction: Tissue factor (TF) expression has been described in various neoplasms and was correlated with angiogenesis and metastases. Objectives: To describe TF expression in colorectal cancers, correlating it with microvessel density and clinical and pathological variables. Methods: Immunohistochemistry was used to determine TF expression and microvessel density. The Student t-test was used to compare high and low TF expression with microvessel density andwith age. The chi-squared test was used for other comparisons, and Kaplan-Meier curves were used for survival analyses. Results: Forty-three patients were operated with curative intent. Their mean age was 58.1±12.6 years old, and 62.8% were male. The rectum was the most common location (60,4%), and most tumors reached the serosa and peri-intestinal fat (72.1%). Lymph nodes were positive in 46.5%, and 72.1% of the tumors were moderately differentiated adenocarcinomas. Death occurred in 27.6±12.8months in 51.1% of the patients who had recurrence. Tissue factor expression was intense in 88.4%. There was a positive correlation between TF expression and microvessel density (p=0.02), and between TF and older age (p< 0.01). There was no correlation between TF expression and other variables (gender, histological type, penetration into the intestinal wall, and lymphatic and systemic metastases). Tissue factor expression did not correlate with survival. Conclusion: Tissue factor expression correlated with increased microvessel density and older age. Further studies are necessary to ascertain the clinical relevance of TF in colorectal cancer. (AU)

The research progress of tissue factor pathway inhibitor / 中华检验医学杂志

Tissue factor pathway inhibitor (TFPI) is a major inhibitor of tissue factor-mediated extrinsic coagulation pathway, mainly derived from microvascular endothelial cells. Recent studies have found that TFPI plays a role in hemophilia, sepsis, antiphospholipid syndrome, venous thromboembolism and other diseases, and participates in the occurrence and development of diseases through anticoagulation mechanism. At present, there are many methods to detect the source, content and function of TFPI, which are helpful for the diagnosis and treatment of clinical diseases.

Molecularly engineered truncated tissue factor with therapeutic aptamers for tumor-targeted delivery and vascular infarction

Selective occlusion of tumor vasculature has proven to be an effective strategy for cancer therapy. Among vascular coagulation agents, the extracellular domain of coagulation-inducing protein tissue factor, truncated tissue factor (tTF), is the most widely used. Since the truncated protein exhibits no coagulation activity and is rapidly cleared in the circulation, free tTF cannot be used for cancer treatment on its own but must be combined with other moieties. We here developed a novel, tumor-specific tTF delivery system through coupling tTF with the DNA aptamer, AS1411, which selectively binds to nucleolin receptors overexpressing on the surface of tumor vascular endothelial cells and is specifically cytotoxic to target cells. Systemic administration of the tTF-AS1411 conjugates into tumor-bearing animals induced intravascular thrombosis solely in tumors, thus reducing tumor blood supply and inducing tumor necrosis without apparent side effects. This conjugate represents a uniquely attractive candidate for the clinical translation of vessel occlusion agent for cancer therapy.

Role of crotoxin in coagulation: novel insights into anticoagulant mechanisms and impairment of inflammation-induced coagulation

Snake venom phospholipases A2 (svPLA2) are biologically active toxins, capable of triggering and modulating a wide range of biological functions. Among the svPLA2s, crotoxin (CTX) has been in the spotlight of bioprospecting research due to its role in modulating immune response and hemostasis. In the present study, novel anticoagulant mechanisms of CTX, and the modulation of inflammation-induced coagulation were investigated. Methods: CTX anticoagulant activity was evaluated using platelet poor plasma (PPP) and whole blood (WB), and also using isolated coagulation factors and complexes. The toxin modulation of procoagulant and pro-inflammatory effects was evaluated using the expression of tissue factor (TF) and cytokines in lipopolysaccharide (LPS)-treated peripheral blood mononuclear cells (PBMC) and in WB. Results: The results showed that CTX impaired clot formation in both PPP and WB, and was responsible for the inhibition of both intrinsic (TF/factor VIIa) and extrinsic (factor IXa/factor VIIIa) tenase complexes, but not for factor Xa and thrombin alone. In addition, the PLA2 mitigated the prothrombinase complex by modulating the coagulation phospholipid role in the complex. In regards to the inflammation-coagulation cross talk, the toxin was capable of reducing the production of the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α, and was followed by decreased levels of TF and procoagulant activity from LPS-treated PBMC either isolated or in WB. Conclusion: The results obtained in the present study recognize the toxin as a novel medicinal candidate to be applied in inflammatory diseases with coagulation disorders.(AU)

RG108 affects the proliferation and apoptosis of NSCLC cells by regulating TFPI-2 methylation and TMPRSS4 expression / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of RG108 on the proliferation and apoptosis of human non-small cell lung cancer (NSCLC) cell lines (A549, H1299) and explore its molecular mechanism. Methods: A549 and H1299 cells were cultured in vitro and treated with different concentrations of RG108. The cell proliferation, cell cycle and apoptosis were detected by MTT assay and Flow cytometry, respectively. qPCR and Western blotting (WB) were used to detect the TFPI-2 mRNA and protein expressions as well as the expression of TMPRSS4 in cells. Meanwhile, the methylation status and degree of TFPI-2 promoter in cells were detected with Methylation-specific PCR (MSP) and colorimetry. Finally, siRNA-TFPI-2 and pcDNA3.0-TMPRSS4 plasmids were used to silence TFPI-2 or overexpress TMPRSS4, and then the changes in cell proliferation and apoptosis were detected. Results: After treatment with RG108, the proliferation rate of A549 and H1299 cells were significantly decreased (all P<0.05), while the apoptosis rate were significantly increased(P<0.05), the cell cycle were arrested in G1/S phase (P<0.05), and the intracellular mRNA and protein expressions of TFPI-2 were significantly increased (P<0.01 or P<0.05). Meanwhile, the methylase degree in TFPI-2 promoter region and the expression of TMPRSS4 in cells were all significanly decreased ( all P<0.05). After TFPI-2 silence, the proliferation levels of A549 and H1299 cells were significantly increased(all P<0.05); however, the apoptosis rate of A549 and H1299 cells were significantly reduced after transfection with pcDNA3.0-TMPRSS4(all P<0.05). Conclusion: RG108 can inhibit proliferation of A549 and H1299 cells and promote apoptosis by inhibiting the methylation of TFPI-2 and negatively regulates the expression of TMPRSS4.

Expressions of tissue factor and vascular endothelial growth factor in diffuse large B-cell lymphoma and their clinical significances / 白血病·淋巴瘤

Objective@#To investigate the expressions of tissue factor (TF) and vascular endothelial growth factor (VEGF) in diffuse large B-cell lymphoma (DLBCL) and their clinical significances.@*Methods@#The clinical data of 80 cases of DLBCL diagnosed at the Second People's Hospital of Lianyungang from January 2010 to December 2017 were collected, and 30 cases of reactive hyperplasia of lymph node (RLN) were selected as the controls. The expressions of TF and VEGF in the two groups were detected by using immunohistochemical staining.@*Results@#The positive rate of TF and VEGF in the DLBCL group was higher than that in the RLN group [TF: 86.3% (69/80) vs. 50.0% (15/30) ; VEGF: 90.0% (72/80) vs. 53.3% (16/30) ; both P < 0.01]. And there was a positive correlation between the expression of TF and VEGF (r = 0.704, P < 0.05). There was no significant difference in the positive rates of TF and VEGF among the patients with different gender, age and Hans subtypes in DLBCL group (all P > 0.05). The positive rate of TF in DLBCL patients with B symptom, increased LDH, physical status grade ≥2, and extranodal lesion number >1 was higher (all P < 0.05). The positive rate of VEGF in patients with Ann Arbor stage Ⅲ-Ⅳ, B symptom, increased LDH, and extranodal lesion number >1 was higher (all P < 0.05). The positive rate of TF in international prognostic index (IPI) high-risk group was higher than that in low-risk group (P < 0.01); the positive rate of VEGF in IPI high-risk group and middle-high-risk group was higher than that in low-risk group (all P < 0.01). The expressions of TF (r = 0.491, P < 0.01) and VEGF (r = 0.529, P < 0.01) were positively correlated with IPI. The overall survival rates of TF and VEGF low-expression group were higher than those of TF and VEGF high-expression group (both P < 0.05).@*Conclusion@#The expressions of TF and VEGF are highly expressed in DLBCL, which is associated with the IPI. It can provide a reference value in evaluating prognosis of DLBCL.

Qilin pills prevent ovarian hyperstimulation syndrome of patients after IVF-ET by regulating plasma-related indicators / 中国临床药理学与治疗学

AIM: To study the preventive effect of Qilin pill on ovarian hyperstimulation syndrome (OHSS) after in vitro fertilization and embryo transfer (IVF-ET) and its effects on vascular endothelial growth factor (VEGF), tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in plasma. METHODS: Sixty-four patients undergoing IVF-ET treated in our hospital from January 2016 to January 2019 were selected. On the day of ovulation induction injection of human chorionic gonadotropin (HCG), 32 patients with high risk factors of OHSS were randomly divided into two groups. The control group received western medicine therapy, while the observation group received extra Qilin pill. The incidence of mild to moderate OHSS, fresh cycle transplant cancellation rate, plasma VEGF, TF, TFPI levels, and clinical outcomes of patients undergoing IVF-ET (HCG positive rate, biochemical pregnancy rate, clinical pregnancy rate) were compared between the two groups.RESULTS:There was no severe OHSS occurred in the two groups, the incidence of OHSS in the observation group (12.50%) and the cancellation rate of fresh cycle transplantation (15.63%) were lower than those in the control group (50.00%, 43.75%)(χ2=6.063,P=0.014); The levels of VEGF and TF in the observation group on the day of egg retrieval and embryo transfer were [(368±103) pg/mL, (392±91) pg/mL],[(24±4)pg/ mL,(29±4) pg/mL], which were lower than the control group [(436±117) pg/mL, (448±108) pg/mL],[(26±4) pg/mL, (31±4) pg/mL] (t=2.450,2.237,4.093,5.204,P=0.017,0.029,<0.001,<0.001); The plasma TFPI levels in the observation group on the day of egg retrieval and embryo transfer were [(73±18) ng/mL,(66±12) ng/mL], higher than the control group [(62±16)ng/mL, (58±10) ng/mL](t=2.550,3.032,P=0.014,0.004); The biochemical pregnancy rate in the observation group (8.70%) was lower than that in the control group (42.86%) (χ2=4.147, P=0.042),the clinical pregnancy rate (91.30%) was higher than that of the control group (57.14%) (χ2=4.147,P=0.042).CONCLUSION:Qilin pill can prevent the occurrence of severe OHSS after IVF-ET, reduce the occurrence of mild to moderate OHSS, decrease the cancellation rate of fresh cycle transplantation and improve the pregnancy outcome after IVF-ET; Its mechanism may be related to the regulation of the expression of VEGF, TF and TFPI.

Research progress of relationship between pancreatic cancer and blood coagulation factors / 复旦学报（医学版）

It has been widely considered that the pancreatic cancer has an inherent and unique ability to induce a hypercoagulable state that leads to clinically significant thrombosis.During examination of lower-limb venous with color doppler blood flow images,more than 50％ of pancreatic cancer patients were found having deep vein thrombosis (DVT).The causes of the hypercoagulable state in pancreatic cancerare still partly understood now.Its relationship with invasion,metastasis and prognosis of pancreatic cancer also need further research.How to deal with this kind of hypercoagulable state is worthy of study.This article discusses changes of some main factors in clotting mechanism of pancreatic cancer.The progress of research on the prevention and treatment is expounded and the future research direction is also put forward.

The effect of Xuebijing injection on the procoagulation of tissue factor by inositol-requiring enzyme 1α signaling pathway / 中华急诊医学杂志

Objective To investigate the effect of Xuebijing injection on lipopolysaccharide (LPS)-mediated the procoagulant activity of tissue factor (TF) in abdominal aortic endothelial cells from rats.Methods Abdominal aortic endothelial cells from rats were randomLy(random number) divided into the control group,LPS group (500 ng/mL),Xuebijing group (1,5,25 μL/mL),and LPS+Xuebijing group (1,5,25 μL/mL),respectively.Cell proliferation was measured by CCK8 and lactate dehydrogenase (LDH) level in supematants was determined at 24,48,and 72 h;Expressions ofinositol-requiring enzyme-1α (IRE 1α),unspliced-box binding protein-1 (uXBP-1),spliced-box binding protein 1 sXBP-1),and protein disulfide isomerase (PDI) were determined by Western blotting at 72 h.Procoagulant activity of TF was measured as the ability of monolayer to support activation of factor with the addition of a and Ca2+ by chromogenic substrate method.Results Compared with the control group,the cell proliferation was decreased and LDH level was increased in the LPS group (P＜0.05),and there were markedly up-regulated in the expression of IRE1 α,uXBP1,sXBP1,and PDI (P＜0.05).Compared with the control group,treatment with Xuebijing injection could promote cell proliferation and reduce the release of LDH (P＜0.05 or P＜0.01),which were gradually enhanced along with the observational intervals.Compared with the LPS group,the LPS+Xuebijing group showed obviously higher cell proliferation and lower release of LDH (P＜0.05 or P＜0.01),expressions of IRE 1 α,uXBP 1,sXBP 1,and PDI were significantly reduced (P＜0.01);meanwhile,F Ⅹ a activity was decreased in the LPS+Xuebijing group,and 5 μ L/mL Xuebijing was the optimal dose in down-regulation of F Ⅹ a.Conclusions These results suggest that treatment with Xuebijing injection can markedly down-regulate the expression of PDI by inhibiting the IRE1α-XBP1 signaling pathway to suppress the procoagulant activity of TF in abdominal aortic endothelial cells from rats.

Changes of microparticle levels before and after daunorubicin-based chemotherapy in patients with acute leukemia / 白血病·淋巴瘤

Objective To observe the serum levels of endothelial microparticles (EMP) and tissue factor-bearing microparticles (TF+MP) in patients with acute leukemia before and after daunorubicin-based chemotherapy. Methods From July 2012 to February 2013, 15 patients with newly diagnosed acute leukemia in Peking Union Medical College Hospital received DA (daunorubicin + cytarabine) regimen or VDCLP (vincristine + daunorubicin + cyclophosphamide + L-asparaginase + prednisone) regimen chemotherapy. There were 8 males and 7 females, and the median age of patients was 44 years old. Eleven patients were acute myeloid leukemia (M01 case, M11 case, M29 cases), and 4 were acute lymphocytic leukemia. The peripheral blood samples were taken before induction chemotherapy and after 3 days of daunorubicin. Levels of EMP and TF+MP were assessed using flow cytometry. Results The serum EMP and TF+MP levels were significantly higher after 3-day daunorubicin infusions than those before induction chemotherapy (28.94/μl vs. 10.74/μl, P= 0.001; 64.24/μl vs. 43.80/μl, P= 0.02). Conclusion Daunorubicin-based chemotherapy may cause increased numbers of EMP and TF+MP in patients with acute leukemia.

Mechanism of coagulation dysfunction in the recipients mediated by tissue factor activation after liver xenotransplantation / 器官移植

Objective To investigate the mechanism underlying the activation of tissue factor (TF) that leads to coagulation dysfunction in the recipients after liver xenotransplantation. Methods Auxiliary heterotopic liver xenotransplantation was performed in 3 minipigs with α-1,3-galactosyltransferase gene-knockout (GTKO) as the donors and Tibetan macaque (Macaca thibetana) as the recipients. Postoperative coagulation function changes in the recipients were observed. Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical staining were adopted to quantitatively measure the expression levels of monkey and minipig TF messenger RNA (mRNA) and protein in the liver tissues of the primary and transplant livers at different time points before and after transplantation. The recalcification time of peripheral blood mononuclear cell (PBMC) was recorded in the normal control monkeys and the recipient monkeys before and 2 h after liver transplantation to evaluate the coagulation status in the recipients. Results All three recipients presented with different degrees of coagulation dysfunction after surgery, manifested as a decrease in fibrinogen level and a reduction in platelet count. The monkey TF protein was positively expressed in the primary livers after surgery, whereas negatively expressed in transplant livers before and after liver transplantation. The minipig TF protein was negatively expressed in both primary livers and transplant livers. At postoperative 2 h, monkey TF mRNA was up-regulated by (2.10±0.24) times in the primary liver compared with the preoperative level, whereas the minipig TF mRNA was up-regulated by (1.42±0.15) times compared with preoperative level. There was statistical significance between the primary livers and transplant livers (P=0.014). Compared with PBMC in the normal control monkeys and recipient monkeys before liver transplantation, the recalcification time of the PBMC in the recipient monkeys was significantly shortened at postoperative 2 h (both P<0.001). Conclusions At the presence of coagulation dysfunction after liver xenotransplantation, the level of TF activation in the primary livers is significantly higher than that in the transplant livers. The TF activation in the primary livers is the main cause of coagulation dysfunction after liver xenotransplantation.

Effect of D-dimer and tissue factor-1208 D/I gene polymorphism on the prognosis of patients with off-pump coronary artery bypass grafting / 中华危重病急救医学

Objective To investigate the effect of perioperative period D-dimer and tissue factor (TF)-1208 D/I gene polymorphism on the long-term prognosis of patients with off-pump coronary artery bypass grafting (OPCABG). Methods Retrospective analysis of the case data of the first OPCABG patients admitted to Tianjin Medical University General Hospital from May 2015 to May 2016 were enrolled. The general data, operation time, bypass number, left ventricular ejection fraction (LVEF), flow rate of 24-hour pleural effusion, intraoperative heparin dosage, combined anticoagulant and antiplatelet time, and the time of postoperative ventilator were measured. The blood biochemical indexes of 1, 4, 7, 14 days and 1, 2, 3 months after operation, perioperative complications, the level of D-dimer in the patients with different TF-1208 D/I gene polymorphism, and prognosis of 1-year follow-up were recorded. The risk factors of recurrent angina 1 year after operation was analyzed by Logistic regression analysis. Results The level of plasma D-dimer was increased continuously after OPCABG, and reached a peak at 1 month after operation [1.94 (1.07, 2.70) mg/L], then decreased, and decreased to preoperative level 3 months after operation [0.20 (0.10, 0.45) mg/L]. The level of D-dimer in TF-1208 I I genotype was significantly higher than that in TF-1208 DD genotype and TF-1208 D/I genotype group at 14 days and 1 month after operation [mg/L: 4.17 (1.54, 5.09) vs. 1.91 (1.07, 2.26), 1.02 (0.91, 1.88) at 14 days; 5.12 (2.41, 6.32) vs. 1.94 (1.18, 2.70), 1.62 (0.22,1.88) at 1 month, all P < 0.05]. The results of 1-year follow-up showed that 25 patients with recurrent angina pectoris without the occurrence of myocardial infarction. The proportion of recurrent angina pectoris in TF-1208 I I genotype was significantly higher than that in TF-1208 DD genotype and TF-1208 D/I genotype group (χ2= 0.197, P = 0.004). Logistic regression analysis showed that LVEF< 0.50 [odds ratio (OR) = 6.482, 95% confidence interval (95%CI) = 1.365-18.763, P = 0.015] and TF-1208 I I genotype (OR = 8.864, 95%CI = 1.613-46.743, P = 0.012) were independent risk factors for recurrent angina pectoris at 1 year after OPCABG. Conclusions After OPCABG, the body was in a hypercoagulable state and lasted for a long time, and almost recovered 3 months after operation. LVEF < 0.50 and TF-1208 I I genotype were independent risk factors of angina pectoris at 1 year after surgery.

Study on the risk factors of chronic obstructive pulmonary disease in acute exacerbation complicated with pulmonary embolism / 国际检验医学杂志

Objective To analyze the risk factors of chronic obstructive pulmonary disease (COPD) in acute exacerbation (AE-COPD)complicated with pulmonary embolism ,and to provide reference for the pathogenesis and treatment .Methods A total of 73 patients with COPD suspected PE admitted to our hospital from May 2015 to April 2016 were enrolled in this study .All patients were examined including WBC ,Neu% ,CRP ,IL-8 ,ESR ,PCT ,ET-1 ,D-dimer ,fibrinogen ,NT-proBNP ,myocardial enzyme ,arterial blood gas ,lactic acid ,CT pulmonary angiography(CTPA)within 48 h of admission .The risk factors of AECOPD with PE or with-out PE were analyzed .Results There were 15 cases with PE ,58 cases without PE in all objects .Neu% ,PCT ,NT-proBNP ,D-di-mer ,LDH ,cTnI ,CRP ,IL-8 ,ET-1 in patients with PE were significant higher than those in patients without PE (P<0 .05) .In the PE group ,the correlation coefficient between CRP and IL-8 was 0 .457(P=0 .087) ,the correlation coefficient between CRP and ET-1 was 0 .598(P=0 .019) ,the correlation coefficient between IL-8 and ET-1 was 0 .695(P=0 .004) .Conclusion Acute exacer-bation of COPD combined with PE is associated with the severity of inflammation in the body ,the more serious the inflammatory re-action ,the corresponding increase in myocardial injury ,the higher the risk of PE .

Study on the risk factors of chronic obstructive pulmonary disease in acute exacerbation complicated with pulmonary embolism / 国际检验医学杂志

Objective To analyze the risk factors of chronic obstructive pulmonary disease (COPD) in acute exacerbation (AE-COPD)complicated with pulmonary embolism ,and to provide reference for the pathogenesis and treatment .Methods A total of 73 patients with COPD suspected PE admitted to our hospital from May 2015 to April 2016 were enrolled in this study .All patients were examined including WBC ,Neu% ,CRP ,IL-8 ,ESR ,PCT ,ET-1 ,D-dimer ,fibrinogen ,NT-proBNP ,myocardial enzyme ,arterial blood gas ,lactic acid ,CT pulmonary angiography(CTPA)within 48 h of admission .The risk factors of AECOPD with PE or with-out PE were analyzed .Results There were 15 cases with PE ,58 cases without PE in all objects .Neu% ,PCT ,NT-proBNP ,D-di-mer ,LDH ,cTnI ,CRP ,IL-8 ,ET-1 in patients with PE were significant higher than those in patients without PE (P<0 .05) .In the PE group ,the correlation coefficient between CRP and IL-8 was 0 .457(P=0 .087) ,the correlation coefficient between CRP and ET-1 was 0 .598(P=0 .019) ,the correlation coefficient between IL-8 and ET-1 was 0 .695(P=0 .004) .Conclusion Acute exacer-bation of COPD combined with PE is associated with the severity of inflammation in the body ,the more serious the inflammatory re-action ,the corresponding increase in myocardial injury ,the higher the risk of PE .

Effects of mangiferin on tissue factor expressionin vascular endothelial cells and the underlying mechanisms / 中国药理学通报

Aim To explore the effects of mangiferin on tissue factor(TF) expression in human umbilical vein endothelial cells(HUVECs) and the underlying mechanisms.Methods HUVECs were isolated and primarily cultured in vitro.After the treatment with mangiferin and oxidized low density lipoprotein(oxLDL), TF expression was determined in HUVECs with real-time PCR and Western blot.Results oxLDLinduced the mRNA and protein expression and pro-thrombotic activity of TF in HUVECs.However, the inductive effects of oxLDL were blocked significantly by mangiferin.Furthermore, mangiferin modified TF expression and activity in a dose-dependent manner.Mangiferin was demonstrated to enhance the activity of peroxisome proliferator-activated receptor gamma(PPARγ).In contrast, GW9662, an antagonist of PPARγ, reversed at least partially the suppressive effects of mangiferin on TF.Conclusion Through activating PPARγ, mangiferin suppresses the expression of TF serving pro-thrombotic functions in endothelial cells.

Puerarin attenuates the expressions ofHUVECs tissue factor and its inhibitor induced by ox-LDL / 基础医学与临床

Objective To investigate the effects of puerarin on the expression of human umbilical vein endothelial cells (HUVECs) tissue factor (TF) and tissue factor pathway inhibitor (TFPI) induced by oxidized low-density lipoprotein (ox-LDL).Methods After HUVECs were incubated with different concentrations of puerarin and 50 mg/L ox-LDL,the expression of TF and TFPI mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blot respectively.Results Compared with control,treatment with ox-LDL caused the augment of TF mRNA and protein expression (P<0.01),and the decrease of TFPI mRNA and protein expression.However,50,100,and 200 μmol/L puerarin blunted the augment of TF mRNA and protein expression and weakened the inhibition of TFPI mRNA and protein expression induced by ox-LDL(P<0.01).Conclusions Puerarin reduces HUVECs TF and TFPI mRNA and protein induced by ox-LDL.

Effects of mTOR activation on anti-β2GPI/β2GPI-stimulated tissue factor expression in THP-1 cells / 临床检验杂志

Objective To investigate the role of mammalian target of rapamycin(mTOR) in the expression of tissue factor(TF) from THP-1 cells induced by β2GPI/anti-β2GPIcomplex.Methods The THP-1 cells were treated with both β2GPI/anti-β2GPI and β2GPI/IgG-APS(β2GPI/IgG from APS patients) complexes.Rapamycin(100 nmol/L),the mTOR inhibitor,was used to exert the intervention experiment.The total RNA and proteins of the THP-1 cells were collected for detection.The mRNA expression level and activity of TF in THP-1 cells were detected by real-time quatitative PCR(RT-qPCR) and TF activity kit respectively.western blotwas used to determine the levels of mTOR and phosphorylated-mTOR(p-mTOR),and p38,p-p38,ERK1/2,p-ERK1/2,JNK,p-JNK,NF-κB p65 and p-NF-κB p65 in THP-1 cells were determined simultaneously.Results Both β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes chould significantly upregulate the mRNA expression and activity of TF,and the phosphorylation levels of mTOR in THP-1 cells(P ＜ 0.05).Rapamycin markedly attenuated the mRNA expression and activity of TF and mTOR phosphorylation induced by β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes (P ＜ 0.05),and also inhibited the phosphorylation levels of p38,ERK1/2 and NF-κB p65 in THP-1 cells induced by β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes (P ＜ 0.05),but did not showed effects on the phosphorylation of c-Jun NH2-terminal protein kinase (JNK) (P ＞ 0.05).Conclusion mTOR could be activated by β2GPI/antiβ2GPI complexes in THP-1 cells and play a crucial role for β2GPI/anti-β2GPI-induced TF expression in THP-1 cells.

Analysis on detection of Hcy ,vWf and TF-PCA within 48 h of onset in patients with coronary heart disease / 国际检验医学杂志

Objective To observe the change of serum homocysteine (Hcy) ,plasma von willebrand factor (vWF) and whole blood tissue factor procoagulant activity (TF-PCA) within 48 h of onset in the patients with coronary heart disease (CHD).Meth-ods The relevant instrument was adopted to detect the level of serum Hcy ,plasma vWF and whole blood TF-PCA in 300 CHD pa-tients and 100 individuals undergoing the healthy physical examination ,and then the statistical analysis was performed.Three hundreds cases of CHD were divided into the stable angina group (SAP group ,n= 96) ,unstable angina group (UAP group ,n=100) and acute myocardial infarction group (AMI group ,n=104).Results The Hcy ,vWF and TF-PCA levels in the CHD patients were higher than those in the control group ,the difference was statistically significant (PUAP group> SAP group ,the difference was statistically significant (P<0.05).The vWF and TF-PCA levels in the AMI group and UAP group were higher than those in the SAP group with statistical difference (P<0.05).The Hcy level in SAP ,UAP and AMI patients complicated with diabetes and hypertension was significantly increased compared with the patients without complicating di-abetes and hypertension ,the difference was statistically significant (P<0.05).The vWF and TF-PCA levels had statistical differ-ence between the UAP group and AMI group(P<0.05).Conclusion Routinely detecting the levels of Hcy ,vWF and TF-PCA has an important clinical value for the diagnosis and curative effect observation in the patients with CHD.

Effects of PDTC and curcumin on expression of tissue factor induced by anti-β2GPI in mice / 中国免疫学杂志

Objective:To investigate whether PDTC or curcumin had effect on anti-β2GPI-induced tissue factor (TF) expression in mice.Methods:BALB/c mice were pretreated with PDTC (100 mg/kg,once a day) by intraperitoneal injection (i.p.) or/and curcumin (50 mg/kg,once a day) by oral gavage for 3 consecutive days at 2 h before 500 μg of anti-β2GPI injections in subsequent experiments.Mouse peritoneal macrophages and aorta were collected,homogenized by sonication.The total RNA and protein were collected from each animal,TF expression was detected by Real-time quatitative PCR and TF activity kit.The phosphorylation of NF-κB p65 and c-Jun/AP-1 was determined by Western blot.Results:Anti-β2GPI cloud significantly upregulate TF expression and phosphorylation of NF-κB p65 and c-Jun/AP-1 in mouse peritoneal macrophages and aorta,compared with NR-IgG treated mice (P＜ 0.05).PDTC or/and curcumin could markedly attenuate anti-β2 GPI-induced TF expression,also inhibit activation of NF-κB p65 and cJun/AP-1 in the aorta and peritoneal macrophages respectively (P＜0.05),but combination of two inhibitors had no synergistic effect.Conclusion:Both PDTC and curcumin could affect the expression of TF induced by anti-β2GPI in mice,indicatiug that PDTC and curcumin has the potential to prevent thrombosis in APS.

Role of PI3K/Akt/eNOS signaling pathway in inhibitory effects of puerarin on ox-LDL-induced TF expression in vascular endothelial cells / 中国病理生理杂志

AIM: To explore the role of phosphatidylinositiol 3-kinase/protein kinase B/endothelial nitric oxide synthase (PI3K/Akt/eNOS) signaling pathways in the inhibitory effects of puerarin on oxidized low-density lipoprotein (ox-LDL)-induced tissue factor (TF) expression in vascular endothelial cells.METHODS: The mRNA expression of TF was detected by real-time fluorescent quantitative PCR.The protein levels of TF and Akt was determined by Western blot.The content of the nitric oxide (NO) was measured by nitrate reduction method.RESULTS: Compared with control group, incubating endothelial cells with ox-LDL significantly induced TF expression at mRNA and protein levels and the dephosphorylation of Akt protein, and decreased NO production.Incubation of the endothelial cells with puerarin for 1 h and then treatment of the cells with ox-LDL decreased the TF expression at mRNA and protein levels, increased Akt protein phosphorylation and intracellular NO content.Co-incubation of the endothelial cells with PI3K inhibitor LY294002 and puerarin for 1 h and then treatment of the cells with ox-LDL augmented the TF expression at mRNA and protein levels and the Akt protein dephosphorylation, and decreased NO production.Co-incubation of the endothelial cells with eNOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) and puerarin significantly decreased the inhibitory effect of puerarin on ox-LDL-induced TF expression at mRNA and protein levels in the endothelial cells, and reduced Akt protein phosphorylation and NO production.CONCLUSION: Puerarin inhibits ox-LDL-induced TF expression at mRNA and protein levels in the human umbilical vein endothelial cells via activation of PI3K/Akt/eNOS signaling pathway.