Cyclin-dependent kinase 5 contributes to apoptosis of vascular endothelial cells during aortic dissection.

Background: Tearing of inner layer of aorta causes aortic dissection (AD), a severe disease with high morbidity and mortality. The pathological development of AD partially results from apoptotic death of aortic endothelial cells (AECs), the trigger and the molecular regulation of which remain largely unknown. Cyclin-dependent kinase 5 (CDK5) was initially detected in the brain as a proline-directed serine/threonine protein kinase regulating neuronal cell cycle re-entry and arrest. The abnormal expression of CDK5 leads to apoptotic cell death following abortive cell cycle re-entry in some neuronal diseases. Although physiological and pathological roles of CDK5 have been widely investigated, the expression and function of CDK5 in AD have not been reported. Therefore, the aim of the present study was to discuss the expression and function of CDK5 in AD. Methods: Gene expression profiles were compared between AD tissues and normal aortic tissues using Gene Expression Omnibus (GEO) database with bioinformatic tools. Different cell types were isolated from the digested AD and normal aortic specimens by fluorescence-activated cell sorting (FACS). Gene expression in cells was quantified by quantitative reverse transcription polymerase chain reaction. Endothelial cells purified by FACS were transfected in vivo with plasmids. Cell growth was analyzed by Cell Counting Kit-8 assay. Cell apoptosis was analyzed by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling assay. Results: Analysis of gene profiles from AD tissues and normal aortic tissues using GEO database showed significant higher expression of CDK5 and its downstream regulated genes, proliferating cell nuclear antigen, cyclin B1, and B-cell lymphoma 2, which are regulators for cell cycle and apoptosis. Analysis of purified cells from AD and normal aortic specimens further confirmed this result and showed that the major source of CDK5 was endothelial cells. Depletion of CDK5 inhibited apoptosis of AECs, while the expression of CDK5 promoted apoptosis of AECs. Conclusions: CDK5 induces apoptosis of AECs to promote AD. CDK5 appears to be a promising novel target for preventing AD.

Gene therapy targeting inflammatory pericytes corrects angiopathy during diabetic wound healing.

Wound healing is impaired in the diabetic status, largely attributable to diabetes-associated angiopathy. Pericytes play critical roles in the stabilization of the formed vessels. The loss and dysfunction of pericytes have been reported in inflammation during diabetes and associated with the pathology of diabetic angiopathy. However, a practical approach that targets inflammatory pericytes to improve diabetic wound healing is lacking. In the current study, we showed that the inflammatory pericytes from wound skin of diabetic patients were impaired in growth potential and underwent oxidative stress and apoptosis. Expression of antioxidant gene oxidation resistance protein 1 (OXR1) specifically in pericytes through an adenovirus carrying OXR1 under a pericyte-specific neuron glia antigen-2 (NG2) promoter (AV-NG2p-OXR1) relieved the oxidative stress, reduced the apoptosis, and recovered the growth potential in diabetic pericytes. Moreover, expression of OXR1 in diabetic pericytes retrieved their potential of both suppressing the migration of co-cultured HUVECs and inducing cell aggregates at the branching points, indicating a functional recovery. In vivo gene therapy using this AV-NG2p-OXR1 to DB/DB mice, the mouse model for type 2 diabetes, significantly improved wound healing, likely through enhancing blood flow at the wound rather than increasing vessel density. Together, our data suggest that gene therapy targeting inflammatory pericytes may improve diabetes-associated impaired wound healing.

Timing of Endovascular Interventions for Iliac Vein Compression Syndrome With Thrombus.

The aim of this study is to explore the timing and method of endovascular intervention for iliac vein compression syndrome (IVCS) with thrombus. Data from 111 patients with IVCS, complicated acute deep vein thrombosis (DVT), or post-thrombotic syndrome (PTS) who underwent endovascular interventions were analyzed retrospectively. Patients were divided into Group A (DVT group), including 56 patients with IVCS and iliofemoral DVT, with or without femoropopliteal DVT, with sudden lower limb swelling, and Group B (PTS group) included 55 patients with IVCS and PTS, including 18 with lower extremity wet ulcers and 32 with lower limb infections. Interventional therapies were used to treat the thrombus and eliminate stenosis and occlusion of the iliac vein. In both groups, clinical symptoms in the lower limbs after surgery were reduced significantly, and PTS incidence was low during long-term follow-up. The cumulative patency rate was 75.2% in the DVT group and 88.6% in the PTS group. Comprehensive interventional therapies are safe and effective in patients with IVCS and thrombi. Long-term efficacy in the PTS group tended to be better than that in the DVT group.

Co-suppression of VEGF-A and VEGF-C inhibits development of experimental hemangioma.

Vascular endothelial growth factor A (VEGF-A) plays a critical role in the development and progression of Infantile hemangioma (IH), the most common vascular tumor occurring during infancy. However, a role of VEGF-C in IH remains unclear. Here, we addressed this question. The expression of VEGF family members in hemangiomas at involuting-phase and at proliferating-phase was compared, by RT-qPCR and by ELISA. VEGF-A and VEGF-C were suppressed by specific short-hairpin interfering RNA (shRNA), respectively. Cell growth was determined in an MTT assay. Cell proliferation was assessed by BrdU incorporation and analysis of cell-cycle regulators by Western blotting. Cell apoptosis was assessed by Annexin V assay and analysis of apoptosis-associated proteins by Western blotting. The effects of VEGF-A suppression, or VEGF-C suppression, or both, on hemangioma growth were analyzed in vivo by bioluminescence assay and by weight of the implanted tumor. Significantly higher levels of VEGF-A and VEGF-C were detected in the proliferating-phase of the hemangiomas than in the involuting-phase of the hemangiomas. Suppression of either VEGF-A or VEGF-C decreased hemangioma cell growth, likely through inhibition of proliferation and enhancement of the apoptosis, while suppression of both VEGF-A and VEGF-C had a more pronounced effect than suppression of either VEGF-A or VEGF-C alone. VEGF-A and VEGF-C seemed to regulate proliferation and apoptosis through different proteins. Suppression of both VEGF-A and VEGF-C had a more pronounced effect than suppression of either one on the growth of the implanted hemangiomas In vivo. Thus, co-suppression of VEGF-A and VEGF-C has better inhibitory effects on the growth of hemangioma.

Midterm Results of Endovascular Treatment for Iliac Vein Compression Syndrome from a Single Center.

BACKGROUND: The efficacy of endovascular interventional treatment for iliac vein compression syndrome (IVCS) is not well studied. The purpose of our study was to investigate the clinical outcome of endovascular interventional treatment for IVCS. METHODS: Data of 68 patients with IVCS, who underwent interventional treatment in our hospital, were analyzed retrospectively. Among these patients, 46 had lower extremity varicose veins. Sixty-five patients underwent stent implantation, and 3 patients underwent simple balloon angioplasty. Fourteen patients had post-thrombotic syndrome (PTS) and 11 patients had acute deep venous thrombosis (DVT). Among these 14 patients with PTS, 12 underwent stent implantation and 2 underwent iliac venous simple balloon angioplasty. On the other hand, 9 of the 11 patients with DVT underwent catheter-directed thrombolysis (CDT) and then stent implantation. Of the remaining 2 patients, one underwent Angiojet Rheolytic thrombectomy (ART) before CDT and the other underwent CDT with simple balloon angioplasty. The stenosis rate of iliac vein and the circumference differences between the affected limb and healthy one were measured before and after operation. These patients were followed up with duplex ultrasound postoperatively. RESULTS: A total of 75 stents were placed in 65 patients. The diameter and the length of stent were 6 to 14 mm (mean 12.5 ± 2.0 mm) and 40 to 260 mm (mean 82.5 ± 36.9 mm), respectively. CDT with/without ART was performed, using urokinase and/or alteplase, after inferior vena cava filtration in all of 11 patients with DVT, without the recurrence of pulmonary embolism. The difference in iliac venous stenosis was statistically significant ([91.2% ± 8.4%] [70.0% ∼ 100.0%] vs. 3.9% ± 13.0% [0 ∼ 70.0%], P < 0.01). The patency rates at 1-, 3-, 6- months, 1 year, and 2 years were 98.5%, 95.6%, 94.1%, 92.4%, and 90.7%, respectively. Four patients (5.9%) suffered from minor bleeding at puncture point and were successfully treated with compression. However, 1 (1.5%) patient underwent stent implantation of right iliac vein that became complicated due to migration of stent to the right ventricle which was successfully arrested by Amplatz Goose Neck Snare Kit. The incidence of PTS was 10.3% (7/68) during the follow-up at 1 to 24 (mean 18.2 ± 7.7) months. CONCLUSIONS: Interventional therapy for patients of IVCS is safe and effective. Satisfactory outcomes were obtained for stent placement for IVCS.

microRNA-143 acts as a suppressor of hemangioma growth by targeting Bcl-2.

Infantile hemangioma is the most common vascular tumor affecting infants, which is associated with clonal expansion of endothelial cells. The aim of this study is to determine the role of microRNA (miR)-143 in the growth and survival of hemangioma-derived endothelial cells (HemECs). We examined the expression of miR-143 in patients with proliferating-phase (n=10) and involuting-phase (n=8) hemangiomas. The effects of ectopic expression of miR-143 on the viability, proliferation, cell cycle distribution, and apoptosis of HemECs were explored. We also identified the target gene(s) that was involved in the activity of miR-143. It was found that proliferating hemangiomas had significantly (P<0.05) lower levels of miR-143 than involuting counterparts. Reexpression of miR-143 significantly reduced the viability and proliferation of HemECs, while knockdown of miR-143 led to an increase in the proliferation of HemECs. Moreover, overexpression of miR-143 arrested HemECs at the G0/G1 phase and promoted caspase-3-dependent apoptosis. At the molecular level, miR-143 overexpression significantly promoted the expression of p21 and p53 and reduced the expression of cyclin D1, CDK2, CDK4, and Bcl-2. Silencing of Bcl-2 phenocopied the effect of miR-143 overexpression on the proliferation and apoptosis of HemECs. Furthermore, co-expression of Bcl-2 reversed the growth-suppressive effect of miR-143 on HemECs. Taken together, miR-143 acts as a suppressor in the growth of HemECs, at least partially, through downregulation of Bcl-2. Reexpression of miR-143 may represent a potential therapeutic strategy for the treatment of proliferating hemangiomas.

[Treatment of thoracolumbar fractures by rehabilitation exercise using knee pads on the orthopedic traction bed].

OBJECTIVE: To discuss the clinical effectiveness in treating thoracolumbar fractures adopting the rehabilitation exercise utilizing knee pads on the orthopedic traction bed. METHODS: From June 1996 to June 2006, we studied the clinical effectiveness of thoracolumbar fractures utilizing knee pads on the orthopedic traction bed for rehabilitation exercise. The cases surveyed total 209, 163 of which had full data. There were 98 males and 65 females with the age from 17 to 74 years (mean, 14.5 years). Consulting time after injury from 30 min to 7 days. Fracture site in T11 had 8 cases, in T12 24 cases, in L1 73 cases, in L2 33 cases, in L3 8 cases, in L4 3 cases, in T12 and L1 14 cases. Compression degree of vertebral anterior border: full compression had 1 case,more than 4/5 had 23, more than 2/3 had 67, more than 1/2 had 40, in 1/3 had 46. RESULTS: Among them, 8 cases with legs paresis no recovery in nerval function or stopping recovery changed methods, and underwent surgical treatment. Others 155 cases were followed up from 2 to 12 years with an average of 3 years and 4 months. The average height of vertebral anterior borders of the 169 injured compressed had increased from 1.55 cm before treatment to 2.70 cm after treatment with an average of 1.15 cm. The height of the injured vertebral anterior borders had recovered from 50.5% (1.55/3.07) before treatment to 89.4% (2.70/3.02) after treatment. Kyphosis angle of the injured vertebral bodies had recovered from 13.25 degrees to -1.6 degrees in average. Twenty-three cases associated with dislocation basic reduction. CONCLUSION: Rehabilitation exercise using knee pads on the orthopedic traction bed can obtain satisfactory clinical effect in treating thoracolumbar fractures, the method is easy. At 3, 7, 10 days after treatment, the height of bed should be adjusted according X-ray.