The aim of the study is to identify the effects and underlying mechanisms of visfatin on inflammation and necroptosis in vascular endothelial cells. Human umbilical vein endothelial cells (HUVECs) were stimulated with visfatin or pretreated with Polyinosinic acid (LOX-1 inhibitor). By using the Western blot, RT-PCR, immunocytochemistry, enzyme-linked immunosorbent assay (ELISA), MTT and flow cytometry technique, the occurrence of inflammation and necroptosis in HUVECs were evaluated. Our results showed that 100 ng/mL visfatin significantly increased the mRNA and protein expression of monocyte chemotactic protein 1 (MCP-1) and LOX-1 after 24 hours’ treatment in HUVECs. However, pretreatment with Polyinosinic acid could significantly reduce the expression of MCP-1 compared with visfatin group. Additionally, 100 ng/mL visfatin could induce the production of necrotic features and increase the mRNA expression of BMF (one of the markers of necroptosis), while pretreating with Polyinosinic acid markedly downregulated the mRNA expression of BMF gene and promoted the cell proliferation. These results indicate that visfatin might induce inflammation and necroptosis via LOX-1 in HUVECs, suggesting that visfatin plays a central role in the development of atherosclerosis.
Venous pressure monitoring is an important indicator for the arteriovenous fistula evaluation. Direct static venous pressure monitoring is recognized as the most accurate way of venous pressure monitoring, which is widely used in the functional monitoring, functional evaluation of arteriovenous fistula, the diagnosis of complications and the evaluation of surgical efficacy. Venous pressure monitoring has obvious advantages and disadvantages, so it is necessary to improve relevant knowledge to correctly guide clinical diagnosis and treatment. In this paper, the study of static venous pressure monitoring of arteriovenous fistula was summarized, in order to improve the understanding of the significance and clinical application of static venous pressure monitoring of arteriovenous fistula.
OBJECTIVE To compare the permeability and incidence rate of complication of arteriovenous internal fistula made by autogenous, homologous, and artificial Teflon blood vessels. METHODS Two hundred and forty one cases with arteriovenous internal fistula made by autogenous, homologous, and artificial Teflon blood vessels were followed up to compare the permeability and incidence rate of complication at 6 months, 1 year, 3 years, and 5 years. RESULTS The incidence rate of complication of autogenous blood vessels was lowest, it had no statistical differences compared with arteriovenous internal fistula made by homologous blood vessels. The permeability of arteriovenous internal fistula made by homologous blood vessels was highest, and it had no statistical differences compared with autogenous blood vessels. The permeability of arteriovenous internal fistula made by artificial Teflon blood vessels was lowest, but the incidence rate of complication was highest, and it had significantly statistical differences compared with arteriovenous internal fistula made by autogenous blood vessels (P lt; 0.01). CONCLUSION Arteriovenous internal fistulas made by autogenous and homologous blood vessels have high permeability and low incidence rate of complication, they are superior to the arteriovenous internal fistula made by artificial Teflon blood vessels.
Objective To investigate the clinical effect and safety of balloon angioplasty (BAP) for patients with venous hypertension associated with autogenous arteriovenous fistula (AVF). Methods Thirty-three patients with venous hypertension associated with AVF were hospitalized between August 2012 and August 2014 in the Department of Nephrology, West China Hospital of Sichuan University. All of the patients received BAP therapy. The clinical characteristics and therapeutic effects were comparatively summarized before and after operation. Results Venous hypertension in all the 33 patients was caused by central venous stenosis or occlusion. Among them, there were 13 cases of stenosis on the site where the left innominate vein crossed the aorta, 10 cases of stenosis at the junction of the left innominate vein and superior vena cava, 4 cases of stenosis at the junction of the left subclavian vein and the innominate vein, 3 cases of right innominate vein stenosis, and 3 cases of innominate vein occlusion. The lesions were most common in the innominate vein. Innominate vein stenosis or occlusion occurred in 19 cases (57.6%), and stenosis at the junction of innominate vein and the superior vena cava or subclavian vein occurred in 14 cases (42.4%). Of the 33 patients, one patient with complete occlusion of the innominate vein did not receive BAP treatment because guide wire could not pass through the occlusion site. The other 32 patients underwent BAP treatment, among whom 30 (93.8%) were successful, and 2 (6.2%) failed. No obvious complications occurred. One day after BAP treatment, patients’ symptoms were significantly relieved. One to seven days later, swelling of the hands, pain, and other symptoms were relieved. In the 30 patients who underwent the treatment successfully, 29 were followed up for 3 to 24 months. Among them, 21 (72.4%) maintained clinical remission and the AVFs were functional, 6 patients (20.7%) got a restenosis later, and 2 patients died. Conclusions BAP is effective and safe for venous hypertension associated with AVF. However, the follow-up of patients has displayed the possible risk of restenosis in long-term outcomes.
ObjectiveTo explore the value of ultrasound evaluation and marking before arteriovenous internal fistula in end-stage renal disease hemodialysis patients. MethodsTwenty-five uremia end-stage patients were admitted into our nephrology department from January 2012 to July 2012. All of the patients had encountered several times of fistula failure or had difficulty in establishing the forearm arteriovenous fistula. We focused on observing the brachial artery, radial artery, cephalic vein, the basilica vein and great saphenous vein. We measured the diameter of the vessels and marked the trend of arteries and veins in the body surface under the ultrasonic navigation. Our goal was to look for appropriate bypass vessels in the elbow and the upper arm. ResultsFourteen patients had endured several times of fistula failure. Among the 14 patients, 9 patients completed the surgery of reengineering fistula and autogenous great saphenous vein transplantation, 2 accepted artificial vascular transplantation, 1 completed the removal of blood clots in the left upper limb artificial blood vessels and arteriovenous internal fistula molding, and 2 gave up surgery. Eleven patients could not complete the arteriovenous fistula operation for the fine forearm superficial vein. Of them, 2 patients accepted artificial vascular operation, 6 underwent autogenous great saphenous vein transplantation, 1 with slender radial artery in diameter completed higher position fistula between the brachial artery and median cubital vein, and 2 gave up surgery. ConclusionArteriovenous internal fistula preoperative ultrasound assessment and marking have very important value in improving the success rate of operation in end-stage uremia patients.
Objective To investigate the effect of keratin 17 (K-17) on the migration, prol iferation and tube formation of human umbil ical vein endothel ial cell (HUVEC), and to real ize the role of K-17 in angiogenesis. Methods After HUVEC were cultured in DMEM medium supplemented with 10%FBS overnight, K-17-siRNA-mixture (experimental group) and Ncontrol-siRNA-mixture (negative control group) were added into HUVEC, respectively, by Lipofectamine 2000 transfection assay, and the final concentration of the siRNA was 50 nmol/L. Lipofectamine 2000 alone was used as the control. After the cells were cultured for 36 hours, the cell prol iferation abil ity was detected by cell counting. After 30-hour culture, the cell’s abil ities of migration and differentiation to tube were detected by 24-well Mill icell units and the collagen gel assay, respectively. In addition, non-siRNA-treated HUVEC were cultured for 24 hours in DMEM medium supplemented with 10%FBS (group A), 2%FBS (group B) and 2%FBS+10 ng/mL bFGF (group C), respectively, and then the expression of K-17 in HUVEC was detected by RT-PCR and Western blot. Results After the treatment with K-17-siRNA for 36 hours, HUVEC exhibited no significant difference in the prol iferation, compared with both control and negative control groups (P gt; 0.05). After transfected with K-17-siRNA for 30 hours, the number of HUVEC in the experimental group which migrated from the upper chamber to the lower chamber of Mill icell wells within 24 hours (3719.0 ± 319.0) was smaller than both control (7 437.5 ± 212.0) and negative control (7 356.3 ± 795.7) groups, with significant difference (P lt; 0.01). However, there was no significant difference between the control group and the negative control group (P gt; 0.05). After HUVEC were transfected with K-17- siRNA for 30 hours, the number of tubes in the experimental group, the negative control group and the control group in 24 hours was (1.1 ± 0.5), (3.6 ± 0.5) and (3.2 ± 0.6) per field, respectively. The experimental group was significantly different from both control and negative control groups (P lt; 0.01), and there was no significant difference between the negative control group and the control group (P gt; 0.05). The expression of K-17 protein in HUVEC in groups A, B and C was 0.25 ± 0.02, 0.08 ± 0.01 and 0.72 ± 0.03, respectively. There was significant difference among these three groups (P lt; 0.01). Conclusion K-17 has no impact on cell prol iferation, but may augment endothel ial cell migration, which may facil itate angiogenesis.
【摘要】 目的 通过比较两种原代人脐静脉内皮细胞的分离培养方法并对细胞特异性抗原进行鉴定,探索提高原代内皮细胞体外培养存活率及纯化率的方法。 方法 采用一次性无菌注射器向人脐静脉灌注消化液,消化液的浓度和消化时间分别025%(质量体积比)胰蛋白酶,10 min和01%(质量体积比)胶原酶Ⅱ,15 min。通过在倒置显微镜下观察细胞的形态特点和用免疫荧光染色的方法对细胞进行鉴定,比较两种消化方法的优劣。 结果 01%胶原酶Ⅱ,15 min的消化方法较025%胰蛋白酶,10 min对原代人脐静脉内皮细胞有更好的分离效果,活细胞数量多且细胞纯度较高。免疫荧光染色结果表明细胞内有Ⅷ因子相关抗原表达。结论 胶原酶Ⅱ可以有效分离脐静脉内皮细胞,最佳消化条件是01%胶原酶Ⅱ,37℃,15 min。【Abstract】 Objective To explore the optimal method for primary culture of human umbilical vein endothelial cells (HUVECs). Methods HUVECs were prepared from human umbilical cords by 01% collagenase Ⅱ digestion for 15 minutes and 025 trypsinase digestion for 10 minutes,respectively. HUVECs were observed under inverted microscope and identified by immunofluorescence.The two methods of digestion were compared. Results More HUVECs were harvested through the method of 01% collagenase Ⅱ for 15 minutes,which expressed Ⅷ related antigen. Conclusion The method of 0.1% collagenase Ⅱ digestion for 15 minutes is a better choice to isolate HUVECs.
ObjectiveTo explore the role of arteriovenous axillary loop graft (AVALG) on chest for establishing hemodialysis access in patients with chronic renal failure. MethodsA retrospective analysis was made on the clinical data of 12 patients with chronic renal failure who underwent an AVALG on chest for hemodialysis access between December 2010 and May 2014. There were 2 males and 10 females with an average age of 65.25 years (range, 46-75 years). The main causes were chronic glomerulonephritis in 6 cases, diabetic nephropathy in 4 cases, and both kidney resection because of urinary tract tumors in 2 cases. The disease duration was 2-12 years (mean, 6 years). The 12 patients all underwent 5-14 times (mean, 7 times) failed prior vascular accesses [arteriovenous fistula (AVF）and arteriovenous graft (AVG)] leading to exhaustion of venous access sites on the upper extremities. ResultsThe AVALG on chest were functionally useful for hemodialysis access, 2-3 times per week, and the blood flow was 250-350 mL/minute; the average time for the first dialysis was 48 days (range, 42-93 days). All patients were followed up 12-54 months (mean, 20.92 months). There was no death during perioperative period. The primary patency rates at 6 and 12 months were 91.7% and 83.3% respectively, and the secondary patency rates at 6 and 12 months were both 100%. After operation, infection (1 case), thrombosis (2 cases), bleeding (2 cases), and swollen (1 case) occurred, which were all cured after corresponding treatment. ConclusionAVALG on chest is a supplementary option for chronic renal failure patients with inadequate upper extremity venous access sites after repeat occlusion.