ABSTRACT
<p><b>OBJECTIVE</b>To transfect pcDNA3.1/hTM plasmids containing human thrombomodulin (hTM) gene into human umbilical vein endothelial cells (HUVECs), and investigate the expression of hTM and anticoagulating function of transfected HUVECs.</p><p><b>METHODS</b>HUVECs were transfected with pcDNA3.1/hTM by lipofectin. Expression of hTM mRNA was determined by semi-quantitative RT-PCR, hTM antigen on HUVECs membrane by immunohistochemistry, and activated protein C (PC) in HUVECs by chronometry. By using a semiautomatic coagulator, the effect of the reacting liquid from transfected HUVECs mixed with PC from normal peripheral blood was assayed.</p><p><b>RESULTS</b>About 10% HUVECs were transfected by pcDNA3.1/hTM with high-level hTM mRNA and protein expression. Activated PC produced by pcDNA3.1/hTM group, pcDNA3.1(+)/neo group and untransfected group was (2.80 +/- 0.43) microg/ml, (0.75 +/- 0.08) microg/ml and (0.85 +/- 0.11) microg/ml, respectively. APTT was (51.68 +/- 2.73) s, (38.38 +/- 2.44) s, (39.65 +/- 2.39) s, (33.93 +/- 1.73) s and (34.60 +/- 1.86) s and PT was (21.89 +/- 1.66) s, (20.56 +/- 1.74) s, (20.42 +/- 2.04) s, (19.57 +/- 1.36) s and (20.16 +/- 1.35) s in pcDNA3.1/hTM group, pcDNA3.1(+)/neo group, untransfected group, inactivating PC group and control, respectively.</p><p><b>CONCLUSIONS</b>The pcDNA3.1/hTM plasmid could be transfected into endothelial cells and expressed biologically functioning hTM protein on HUVECs membrane. Activated PC could inhibit intrinsic coagulation pathway obviously with slight effect on extrinsic pathway.</p>
Subject(s)
Humans , Cells, Cultured , Endothelial Cells , Metabolism , Endothelium, Vascular , Cell Biology , Gene Expression Regulation , Genes, Transgenic, Suicide , In Vitro Techniques , Partial Thromboplastin Time , Plasmids , Protein C , Prothrombin Time , Thrombomodulin , Genetics , Transfection , Umbilical Veins , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the surgical methods and the outcome of management for traumatic arterial aneurysm (TAA) and traumatic arteriovenous fistula (TAVF).</p><p><b>METHODS</b>A total of 121 patients with TAA or TAVF were treated by surgery. Clinical, operative and postoperative data were collected and analyzed retrospectively.</p><p><b>RESULTS</b>The surgical techniques included aneurysmectomy and arterial end-to-end anastomosis or vascular grafting or artery ligation, aneurysm ligation and bypass, vascular repair, fistula excision and vascular ligation or vascular grafting or repair and so on. One patient died (0.83%). The follow-up rates of TAA and TAVF were 65.7% and 60% respectively.</p><p><b>CONCLUSIONS</b>Complicated TAA and TAVF in different sites should be treated with different methods.</p>