Safety and Efficacy of Low Dosage of Urokinase for Catheter-directed Thrombolysis of Deep Venous Thrombosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Catheter-directed thrombolysis (CDT) has been a mainstay in treating deep venous thrombosis (DVT). However, the optimal dosage of a thrombolytic agent is still controversial. The goal of this study was to evaluate the safety and efficacy of low dosage urokinase with CDT for DVT.</p><p><b>METHODS</b>A retrospective analysis was performed using data from a total of 427 patients with DVT treated with CDT in our single center between July 2009 and December 2012. Early efficacy of thrombolysis was assessed with a thrombus score based on daily venography. The therapeutic safety was evaluated by adverse events. A venography or duplex ultrasound was performed to assess the outcome at 6 months, 1 year and 2 years postoperatively.</p><p><b>RESULTS</b>The mean total dose of 3.34 (standard deviation [SD] 1.38) million units of urokinase was administered during a mean of 5.18 (SD 2.28) days. Prior to discharge, Grade III (complete lysis) was achieved in 154 (36%) patients; Grade II (50-99% lysis) in 222 (52%); and Grade I (50% lysis) in 51 (12%). The major complications included one intracranial hemorrhage, one hematochezia, five gross hematuria, and one pulmonary embolism. Moreover, no death occurred in the study.</p><p><b>CONCLUSIONS</b>Treatment of low-dose catheter-directed thrombosis is an efficacious and safe therapeutic approach in patients with DVT offering good long-term outcomes and minimal complications.</p>

Stenting of iliac vein obstruction following catheter-directed thrombolysis in lower extremity deep vein thrombosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Catheter-directed thrombolysis (CDT) for deep venous thrombosis (DVT) of the lower extremity has good effect, but whether iliac vein stent placement after thrombolytic therapy is still controversial. The goal of this study was to evaluate the efficacy of stent placement in the iliac vein following CDT in lower extremity DVT.</p><p><b>METHODS</b>This was a single-center, prospective, randomized controlled clinical trial. After receiving CDT, the major branch of the distal iliac vein was completely patent in 155 patients with lower extremity DVT, and 74 of these patients with iliac vein residual stenosis of >50% were randomly divided into a control group (n = 29) and a test group (n = 45). In the test group, stents were implanted in the iliac vein, whereas no stents were implanted in the control group. We evaluated the clinical indicators, including patency of the deep vein, C in CEAP classification, Venous Clinical Severity Score (VCSS), and Chronic Venous Insufficiency Questionnaire (CIVIQ) Score.</p><p><b>RESULTS</b>All patients had postoperative follow-up visits for a period of 6-24 months. Venography or color ultrasound was conducted in subjects. There was a significant difference between the patency rate at the last follow-up visit (87.5% vs. 29.6%) and the 1-year patency rate (86.0% vs. 54.8%) between the test and control groups. The change in the C in CEAP classification pre- and post-procedure was significantly different between the test and control groups (1.61 ± 0.21 vs. 0.69 ± 0.23). In addition, at the last follow-up visit, VCSS and CIVIQ Score were both significantly different between the test and control groups (7.57 ± 0.27 vs. 0.69 ± 0.23; 22.67 ± 3.01 vs. 39.34 ± 6.66, respectively).</p><p><b>CONCLUSION</b>The stenting of iliac vein obstruction following CDT in lower extremity DVT may increase the patency of the deep vein, and thus provides better efficacy and quality of life.</p>

Rapamycin and 3-methyladenine regulate apoptosis and autophagy in bone-derived endothelial progenitor cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mammalian target of rapamycin (mTOR) is involved in a caspase independent form of programmed cell death called autophagy. The aim of this research was to investigate the effects of rapamycin and 3-methyladenine (3-MA) on autophagy, proliferation, apoptosis, and cell-cycle parameters of rat bone marrow-derived endothelial progenitor cells (EPCs).</p><p><b>METHODS</b>Mononuclear cells isolated from rat bone marrow were treated with rapamycin (0.01, 0.1, 1, or 10 µg/L) or 3-MA (1.25, 2.5, 5, or 10 mmol/L) for 24 hours. Expression of the autophagy marker protein LC3-II was analyzed by Western blotting. Apoptosis and cell-cycle progression were analyzed by flow cytometry. Cell proliferation was measured using the MTT assay.</p><p><b>RESULTS</b>Rapamycin treatment of EPCs induced apoptosis and autophagy and inhibited proliferation and cell-cycle progression in a dose-dependent manner. Treatment with 5 mmol/L 3-MA promoted cell proliferation; in contrast, treatment with 10 mmol/L 3-MA promoted apoptosis and induced S-phase arrest.</p><p><b>CONCLUSIONS</b>Rapamycin treatment of EPCs induced apoptosis and autophagy. Low concentrations of 3-MA had no significant effect on the proliferation and apoptosis of EPCs; The 5 mmol/L group promoted cell proliferation, but had no effect on the apoptosis; the 10 mmol/L group inhibited the proliferation and promoted apoptosis through the cell cycle.</p>

Endovascular treatment of iliac vein compression syndrome / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Iliac vein compression syndrome (IVCS), the symptomatic compression of the left common iliac vein between the right common iliac artery and the vertebrae, is not an uncommon condition. The aim of this research was to retrospectively evaluate long-term outcome and the significance of endovascular treatment in patients with left IVCS.</p><p><b>METHODS</b>Between January 1997 and September 2008, 296 patients received interventional therapy in the left common iliac vein. In the second stage, 170 cases underwent saphenous vein high ligation and stripping. Two hundred and thirty-one cases were followed up over a period of 6 to 120 months (average 46 months) and evaluated for symptom improvement with color ultrasound and ascending venography.</p><p><b>RESULTS</b>The stenotic or occlusive segments of the left iliac vein were successfully dilated in 285 cases, of whom 272 received stent implantation therapy. Most of the patients achieved satisfactory results on discharge. During the follow-up period, varicose veins were alleviated in 98.7% of the patients, and leg swelling disappeared or was obviously relieved in 84% of cases. About 85% of leg ulcers completely healed. The total patency rate was 91.7% as evaluated with color ultrasound and 91.5% with ascending venography.</p><p><b>CONCLUSIONS</b>Endovascular treatment of IVCS provides effective symptomatic improvement and good long-term patency in most patients.</p>

Transplantation of VEGF165-gene-transfected endothelial progenitor cells in the treatment of chronic venous thrombosis in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The organization and recanalization of thrombi is a dynamic and complex process. The aim of this research was to study the cotherapeutic effect of stem cell transplantation and gene transfection on chronic venous thrombosis.</p><p><b>METHODS</b>We constructed a recombinant adenoviral vector carrying the vascular endothelial growth factor 165 (VEGF165) gene by using the pAdEasy system, which was subsequently identified and amplified. Simultaneously, endothelial progenitor cells (EPCs) were isolated from rat bone marrow using Ficoll, cultured in EBM-2MV medium, and identified. Then, the cells were transfected with the recombinant Ad-VEGF165. The EPCs were labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (Dil) before transplantation. A rat model of chronic vein thrombosis was developed by partial ligation of the inferior vena cava. The rats were randomly divided into 4 groups (n = 25, each): A, Ad-VEGF165/EPC-transplantation group received 1 ml (10(6)) of Ad-VEGF165/EPCs; B, EPC-transplantation group received 1 ml (10(6)) of EPCs; C, Ad/EPC-transplantation group received 1 ml (10(6)) of Ad/EPCs; D, control group received 1 ml of the transplantation medium. The thrombi and adjacent caval walls were harvested 28 days after transplantation; real-time quantitative polymerase chain reaction was used to detect the expression level of vascular endothelial growth factor (VEGF) mRNA; and western blotting was used to measure changes in VEGF protein expression. Hematoxylin-eosin staining and immunohistochemical staining were performed to detect recanalization. Neovascularization was detected by immunohistochemical staining using the antibody for von Willebrand factor (vWF), which is a component of endothelial cells. The capillary density was quantitatively determined by counting the capillaries under a high-power microscope.</p><p><b>RESULTS</b>The Ad-VEGF165 was constructed, and bone-marrow-derived EPCs were cultivated and successfully identified. We determined the optimum transfection ratio that promoted the growth of EPCs. After transfection, the EPCs secreted the VEGF protein. After transplantation, the in vivo survival of EPCs and their differentiation into endothelial cells were determined by detecting the fluorescence associated with the Dil stain. VEGF mRNA was expressed in groups A, B, C and D after transplantation, and the VEGF mRNA level in group A was significantly higher than those in groups B, C and D (P < 0.05); the VEGF mRNA levels in groups B and C were significantly higher than those in group D (P < 0.05), and there was no statistical significance between the VEGF mRNA levels in groups B and C. The recanalization capillary density in group A was significantly higher than those in groups B, C (P < 0.05) and D (P < 0.01); the recanalization capillary densities in groups B and C were significantly higher than that in group D (P < 0.05). Moreover, there was no statistical significant difference between the values for groups B and C.</p><p><b>CONCLUSIONS</b>The EPCs were successfully transfected by Ad-VEGF165. A suitable transfection ratio can improve the efficiency of EPCs and the possibility of promotion of angiogenesis after transplantation. Transfected EPCs caused accelerated organization and recanalization of vein thrombi.</p>

Radical correction of Budd-Chiari syndrome / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Interventional therapy is widely accepted as the first choice for the treatment of the Budd-Chiari syndrome, but the use of radical correctional therapy should not be discarded. This study describes radical correction by controlling bleeding from distal end of pathological segment of the inferior vena cava (IVC) and discusses potential surgical errors and postoperative complications.</p><p><b>METHODS</b>Of the 216 patients in the study, 78 were treated with simple membranectomy, 64 with dissection of the pathological segment of the IVC and vascular prosthesis or pericardial patch plasty, 60 with resection of the pathological segment of the IVC and orthotopic graft transplantation with vascular prosthesis, and 14 with resection of the occlusive main hepatic vein and its upper IVC, hepatic venous outflow plasty and vascular prosthesis orthotopic graft transplantation from the hepatic venous entrance to the IVC of right atrial ostium.</p><p><b>RESULTS</b>Except 14 cases who were discharged after hepatic vein outflow plasty, four cases died postoperatively, and 198 patients were discharged without complications. The symptoms of 15 patients were relieved partially and 2 without any change. There were no deaths intraoperatively. Of the 112 cases who were followed up for 72 months, 13 suffered from a relapse.</p><p><b>CONCLUSIONS</b>Radical correction is a beneficial therapy in the treatment of Budd-Chiari syndrome.</p>

Effects of bone marrow-derived endothelial progenitor cell transplantation on vein microenvironment in a rat model of chronic thrombosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Endothelial progenitor cells (EPCs) have been used in both experimental studies and clinical treatments of limb ischemia, as well as in the construction of engineered vascular tissue. The objective of this study was to investigate the effects of transplanted bone marrow-derived EPCs on the vein microenvironment in a rat model of chronic vein thrombosis.</p><p><b>METHODS</b>Mononuclear cells were isolated from the bone marrow of immature rats by density gradient centrifugation, cultured, and then transplanted into experimentally induced thrombi into inferior vena cava through the femoral vein. Vascular endothelial growth factor (VEGF), angiopoietin-1 (ANG-1) and monocyte chemotactic protein-1 (MCP-1) mRNA and protein expression levels were measured by real-time quantitative polymerase chain reaction and Western blotting of thrombi and adjacent caval walls 28 days post-transplantation.</p><p><b>RESULTS</b>Levels of VEGF, ANG-1, and MCP-1 mRNA in EPC-transplanted thrombi were 100%, 230.7%, and 212.5% of levels detected in the sham-operated group (P < 0.01), and 99.9%, 215.4%, and 177.8% of levels detected in the experimental control group (P < 0.01). VEGF, ANG-1 and MCP-1 protein levels exhibited a similar trend.</p><p><b>CONCLUSIONS</b>Transplanted bone marrow-derived EPCs appear to alter the vein microenvironment in experimentally induced chronic vein thrombosis by upregulating cytokines associated with thrombic organization and recanalization.</p>

An in vitro study of PcDNA3.0-hVEGF165 transfection on endothelial progenitor cell derived from murine bone marrow / 中国普通外科杂志

Objective To investigate the eftect of VEGF gene transtection on endothelial progenitor cell derived from murine bone marrow.Methods Wistar rat's bone marrow was obtained, mononuclear cell isolated,and endothelial progenitor cells(EPS)were cultured in EGM-2MV.EPCs were identified by immunocytochemistry and electron microscope.EPCs were transfected by liposome mediated pcDNA3.0-hVEGF165.VEGF protein level was determined in the cultural medium supernatant after VEGF transfection by ELISA.Cultural medium supernatant was used to co-culture with ECV304,VEGF protein activity was evaluated by MTT.EPCs expression of vWF,VEGF,FLK-1 was detected by immunocytochemistry.Results EPCs were effectively enriched by EGM-2MV,and the EPCs obtained express the typical cell surface markers such as CD34,CD133,FLK-1.The concentration of VEGF protein in supernatant reaches 1280 pg/ml in the 7th day after pcDNA3.0-hVEGF transfection.No influence of EPCs proliferation could be found after transfeetion.The cell surface marker expression of VEGF,FLK-1, vWF became higher with time,and the ratios of positive cell were 88.52%,82.65% and 95.97% respectively.Conclusions pcDNA3.0-hVEGF165 transfeet EPCS mediated by liposome could excrete a high concentration of functional VEGF protein.It is helpful for EPC to maintain the characters of endothelial cell after VEGF gene transfection and differentiate to mature endothelial cell.