[Expression of fusion protein encoding EGFP-EGF1 of rat coagulation factor VII and its binding function].

This study was aimed at clarification of the function of EGF(1) segment in rat coagulation factor VII with tissue factor (TF) by means of the expression of the fusion protein of EGFP-EGF(1). The DNA fragment encoding EGF(1) was amplified from a rat liver tissue by RT-PCR, and then inserted in an EGFP-procaryotic expression vector to construct the recombinant plasmid pET28a-EGFP-EGF(1) which was introduced into the competent cells of E.coli BL21, then an engineering bacteria strain was obtained which was induced by IPTG to express the fusion protein of EGFP-EGF(1).

[Inhibition of tissue factor by siRNA enhances doxorubicin-induced apoptosis in human neuroblastoma].

Objective: To investigate the regulation of tissue factor (TF) on doxorubicin-induced apoptosis in human neuroblastoma.

Method: The expression of TF was examined by Western blotting. TF siRNA-pSUPER plasmid was constructed by inserting a specific 19-nt silencing sequence targeting TF gene into pSUPER vector.

[Effect of arsenic trioxide on the expression of apoptosis-related genes in NB4 cells].

The aim of this study was to investigate the gene expression profiles of acute promyelocytic leukemia (APL) cell line NB4 treated with arsenic trioxide (As2O3) by using cDNA microarray. cDNA probes were prepared through reverse transcription from mRNA of NB4 cells treated with or without arsenic trioxide. The probes were labeled with Cy3 and Cy5 fluorescence dyes individually, hybridized with cDNA microarray representing 201 different human genes, and their fluorescent intensities were scanned.

[Regulation of tissue factor expression by anti-oncogene PTEN in human neuroblastoma cell line].

Objective: To explore the role of PTEN gene in the regulation of tissue factor (TF) expression in human neuroblastoma cells.

Methods: Expression of PTEN or TF was determined by Western blotting. Transcription of TF was examined by RT-PCR.

Identification of brain-derived neurotrophic factor as a novel angiogenic protein in multiple myeloma.

Patients with multiple myeloma (MM) have increased bone marrow angiogenesis, but the angiogenic properties of myeloma cells and the mechanism of MM-induced angiogenesis have not been completely clarified. The brain-derived neurotrophic factor (BDNF) and its high-affinity receptor, TrkB, have been identified as critical factors in the regulation of vessel formation. In this study, we demonstrate that patients with MM had increased BDNF and vascular endothelial growth factor (VEGF) in their peripheral blood.

Brain-derived neurotrophic factor promotes growth and migration of multiple myeloma cells.

The evolution of multiple myeloma (MM) depends on complex signals from the bone marrow microenvironment, which support proliferation and survival of malignant plasma cells. Previous study defined a brain-derived neurotrophic factor-tyrosine kinase receptor B (BDNF/TrkB) axis in myeloma and autocrine growth stimulation by BDNF in various tumor cells. We examined the biological effects of BDNF on MM cells.

[Regulation of tissue factor expression in brain microvascular endothelial cells by PLA nanoparticles coating NF-kappaB decoy oligonucleotides].

Objective: To investigate a new strategy of polylactic acid (PLA) nanoparticles delivery system coating nuclear factor-kappaB (NF-kappaB) decoy oligonucleotides (ODNs) for inhibiting TF expression in cultured brain microvascular endothelial cells(BMECs).

Methods: PLA nanoparticles coating FITC-labeled NF-kappaB decoy ODNs were formulated by nano-deposition method and the characteristics of nanoparticles were detected. BMECs were isolated and cultured in vitro.

[Factor VII detection and its association with lipid in ischaemic heart disease].

Objective: To study the variation and significance of plasma factor VII (FVII) in various types of ischemia heart disease (IHD) and discuss its relation with plasma lipid.

Methods: FVIIa was determined with one stage clotting assay using a recombinant soluble tissue factor (rsTF). FVIIc was measured with one stage clotting assay.

[Regulatory effect of IL-10 on expression of tissue factor induced by IL-6 in peripheral blood mononuclear cells].

To investigate the role of anti-inflammatory cytokine in acute coronary syndrome (ACS), the effect of IL-10 on expression of tissue factor (TF) induced by IL-6 in peripheral blood mononuclear cells (PBMNC) were studied. PBMNC were allowed to culture with rhIL-10 before being stimulated by rhIL-6. One-step recalcification clotting time was used to evaluate procoagulant activity (PCA) of PBMNC.

[Study on the high expression of brain-derived neurotrophic factor in multiple myeloma patients and its possible mechanism].

In order to investigate the expression of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) in multiple myeloma patients and the in vitro and in vivo proangiogenic effects of BDNF, the plasma concentrations of BDNF and VEGF in MM patients and control group were determined by ELISA, the effect of BDNF on the in vitro proliferation of human umbilical vein endothelial cells (HUVEC) was examined by MTT assay; the effects of BDNF on HUVEC migration and tube formation were studied by modified Boyden chamber assay and tube formation assay, respectively. Matrigel plug assay and chorioallantoic membrane assay were used to evaluate the effect of BDNF on angiogenesis in vivo. The results demonstrated that the concentration of BDNF was (4.

[Coagulation factor VII levels in uremic patients and theirs influence factors].

This study was aimed to investigate coagulation factor VII level in uremic patients with chronic renal failure and to explore theirs influence factors. The plasma levels of coagulation factor VII were detected in 30 uremic patients with chronic renal failure before and after hemodialysis for 1 month, the factor VII activity (FVII:C) was determined by one-stage coagulation method, while activated factor VII (FVIIa) was measured by one-stage coagulation method using recombinant soluble tissue factor, and factor VII antigen was detected by ELISA. The results showed that: (1) The FVIIa, FVII:C and FVIIAg levels in chronic uremic patients before hemodialysis were 4.

[The effects of tissue factor/activated factor VII complex on the invasion and metastasis of human ovarian cancer].

Objective: To explore the role of tissue factor/activated factor VII (TF/FVIIa) complex in human ovarian cancer invasion and metastasis.

Methods: (1) Constructed an expression vector of TF, pcDNA3-TF and established a human ovarian cell line A2780/TF expressing high level TF by using molecular cloning and gene transfection techniques. (2) By Boyden chamber assay to count the numbers of A2780 and A2780/TF cells that penetrated the matrigel to the back of PVPF membrane after FVIIa stimulation.

[Study of the effect of TF/FVIIa complex on the expression of u-PAR mRNA in human ovarian cancer].

Objective: To construct the expression vector of human tissue factor (TF), and investigate the influence of TF/coagulant factor VIIa (FVIIa) complex on the transcriptional expression of urokinase plasminogen activator (u-PA) and u-PA receptor (u-PAR) in human ovarian cancer.

Methods: The human TF cDNA was obtained from placenta by RT-PCR and then inserted into eukaryotic expression vector pcDNA3 to obtain the TF-pcDNA3 combinant. This combinant was transfected into human ovarian cancer cell line A2780 by lipofectamine.

[Detection of thrombus precursor protein and its clinical significance].

Objective: To discuss the significance of the concentration change of plasma thrombus precursor protein (TpP) in the diagnosis of disseminated intravascular coagulation (DIC) or pre-DIC.

Methods: Enzyme linked immunosorbent assay (ELISA) was used in the detection of plasma TpP in patients with different diseases (77) and normal adults (47). At the same time, other correlated markers were detected and statistical comparison was proceeded.

[Cloning of expression vector of human tissue factor gene and its expression in human ovarian cancer cell line].

The aim was to construct the expressive vector of human tissue factor (TF), and determine its expressive level in stable-transfected human ovarian cancer cell line. The human TFcDNA was obtained from human placenta by RT-PCR and then inserted into eukaryotic expressive vector pcDNA3 to obtain the TF-pcDNA3 recombinant. This recombinant gene was introduced into human ovarian cell line A2780 through transfection mediated by lipofectamine.

[Tissue factor expression in human umbilical vien endothelial cells stimulated by TNF-alpha and its molecular mechanism].

The objective of this study was to explore tissue factor (TF) expression induced by TNF-alpha in cultured human umbilical vien endothelial cells (HUVEC) and its molecular mechanism. TF expression on the surface of HUVEC, TF mRNA and nuclear factor kappaB (NF-kappaB) in HUVEC were detected by flow cytometry, RT-PCR and Western blot respectively. The results showed that TNF-alpha could enhance TF expression on the surface of HUVEC, the TF expression increase was highly consistent with the increased synthesis of TF mRNA, and the increase of TF expression was lately appeared for several hours.

[Study of the inhibitory effect of NF-kappaB decoy on tissue factor gene expression and FVII activation in cultured human umbilical vein endothelial cells].

Objective: To investigate the inhibitory effect of NF-kappaB decoy on tissue factor (TF) expression and FVII activation in cultured human umbilical vein endothelial cells (HUVEC), and to explore new methods for prevention and treatment of coronary heart disease.

Methods: NF-kappaB decoy transfection efficiency was detected by flow cytometry, NF-kappaB decoy's mechanism was analyzed by electrophoretic mobility shift assay (EMSA), TF mRNA was detected by RT-PCR, TF antigen expression on the surface of HUVEC by flow cytometry, FVIIa level in plasma incubated with HUVEC stimulated by TNF-alpha by rsTF one stage clotting method.

Results: NF-kappaB decoy could be successfully transfected into HUVEC.

[Experimental study on activating antileukemic T cells by vaccination with dendritic cells pulsed with survivin].

The objective of this study is to investigate the effect of vaccination with dendritic cells pulsed with survivin antigen on activation of antileukemic T cells, and inhibiting proliferation of leukemic cells. The expression of survivin on acute leukemic cells were detected by cofocal microscopy and immunoprecipitation-Western blot. DCs collected from peripheral blood mononuclear cells were pulsed with survivin purified proteins.

[Construction of expression vector for recombinant annexin II and characteristics of its fibrinolysis].

The study was designed to investigate annexin II resulting in molecular pathological mechanism of the primary fibrinolysis and establish annexin II vector model for further research on disturbance of coagulation. A target gene was amplified from human umbilical vein endothelial cells (HUVEC) by RT-PCR. Annexin II gene fragment was purified and ligated with molecular biological recombinant technology.