Objective To induce embryonic stem cell (ESC) to differentiate into endothel ioid cells using a simple adhesive culture method, and to provide a new cells seed source for vascular tissue engineering or cell therapy. Methods SV129-derived ESC were seeded at 2 × 104/cm2 and maintained undifferentiated on ESC culture medium in the presence of 1 000 U/mL leukaemia inhibitory factor (LIF). Embryoid body (EB) formatted when ESC cultured in suspension in the lack of LIF. At 4 days, EB was transferred to 0.1% gelatin coated dish and cultured with medium supplementary of VEGFto be induced differentiation. The characteristics of differentiated cells were determined by immunohistochemistry staining, flow cytometry (FCM), 1, 1-dioctadecyl-3, 3, 3, 3-tetramethyl indocarbocyanine-labeled acetylated low density l ipoprotein (DiIAc- LDL) takeup test, and TEM detection. Results Differentiated cells were morphologically characterized as endothel ial cells. They could takeup DiI-Ac-LDL, be stained positive by Flk-1 and CD31. The CD31 positive cells reached above 90% when measured by FCM. Furthermore, Weibel-Palade bodies were detected and tight junctions were found when differentiated cells were examined by TEM. Conclusion Using a simple adhesive culture method and by suppl ied with VEGF alone, ESCs can be induced to differentiate into endothel ioid cells. The differentiation method is simple and economic, and can provide seed cells for vascular tissue engineering or cell-therapy.
ObjectiveTo summarize the feasibility and effectiveness of endovascular treatment for vertebral artery stenosis ectopic originating from the aortic arch. MethodThe clinical data of 5 patients with vertebral artery stenosis ectopic originating from aortic arch admitted to Xuanwu Hospital of Capital Medical University from January 2020 to May 2023 were retrospectively analyzed. ResultsAll 5 patients underwent magnetic resonance angiography and CT angiography before treatment. The vertebral arteries of all patients originated from the aortic arch and were severe stenosis, with a median diameter of 1.3 mm (range, 1.1–1.5 mm) by CT angiography. All 5 patients were successfully treated with endovascular intervention. The operation time was 37–45 min, with a median of 39 min. There were no complications of vertebral artery rupture, embolism or occlusion, cerebral infarction, plaque abscission, hematoma at puncture site, pseudoaneurysm, postoperative hemorrhage, or perioperative pulmonary infection. The patients were followed-up for 13–30 months, with a median follow-up time of 20 months. The blood vessels of 5 patients remained unobstructed and no clinically significant vascular restenosis occurred. ConclusionsBased on the experiences of 5 cases of vertebral artery stenosis ectopic originating from the aortic arch, it is safe and effective to choose endovascular treatment for severe symptomatic vertebral artery stenosis. Especially, it is necessary to evaluate the stenosis degree of vertebral artery using CT angiography before treatment.
Objective To explore the mechanism of mesenchymal stem cells （MSCs） transplantation for chronic hindlimb ischemia in Lewis rats by using cell tracer technique. Methods MSCs were isolated and cultured by using density gradient centrifugation and adherence method respectively， then labeled by 5-bromo-2-deoxyuridine （BrdU）. Eight chronic hindlimb ischemia models of Lewis rats were prepared by using suture-occluded method and then divided randomly to MSCs transplantation group and control group， each group enrolled 4 rats， accepting MSCs transplantation and saline respectively. Then on 7 days and 14 days after transplantation， clinical observation， determination of blood flow， and angiography were performed on rats of the 2 groups. At the same time points after previous tests， rats of the 2 groups were sacrificed to get quadriceps tissues and gastrocnemius tissues to perform HE staining and BrdU immunohis-tochemistry. Results The 8 rats were all survived on 14 days after transplantation， with no tumor happened and necroses in the transplanted area. On 14 days after transplantation， the blood flow ratio of operated side to un-operated side in the hindlimb （1.773 vs. 1.279） of rats in MSCs transplantation group and control group increased， and the angiography results showed that there were no much increase in ratio of collateral vessels number （0.908 vs. 0.835）. There were no significant change in the quadriceps tissues and gastrocnemius tissues by HE staining. The BrdU positive kernels located in the inter-stitial substance cells and vascular endothelia cells， and divided differently in different parts of hindlimb at different time points， that the ratio of positive cells in gastrocnemius tissue was higher than those of quadriceps tissue on 7 days after transplantation， but lower on 14 days. Conclusions MSCs transplantation can increases the blood perfusion of hindlimb in the early stage of chronic hindlimb ischemia model， and the possible mechanism may be the paracrine effect of MSCs but not the number increase of collateral vessels.
Objective To explore the effective surgical approaches in treating subclavian artery occlusion. Methods Between December 2005 and February 2010, 53 patients with subclavian artery occlusion were treated, including left subclavian artery occlusion (35 cases) and stenosis (5 cases), right subclavian artery occlusion (5 cases) and stenosis (4 cases), and bilateral subclavian artery occlusion (4 cases). There were 40 males and 13 females with an average age of 64 years (range, 22-77 years), including 49 cases of arteriosclerosis obl iterans and 4 cases of aortic arteritis. The disease duration was 15 days to 20 months (6.5 months on average). In 49 patients with unilateral subclavian artery occlusion, 39 cases compl icated by carotid or / and cerebral artery lesion underwent axillo-axillary bypass grafting, and 10 cases without carotid or /and cerebral artery lesion underwent carotid-subclavian bypass grafting. Ascending aorta to bisubclavian bypass graftings were performed on 4 cases with bilateral subclavian artery occlusion. After operation, patients received routine treatment with anticoagulant and antiplatelet agents. Results The operations were successfully performed in 52 cases with a successful rate of 98.11%. Thrombogenesis at anastomotic site occurred in 1 case of aortic arteritis after 48 hours. Two cases had brachial plexus crush injury and 4 had hematoma around the bilateral anastomosis after axillo-axillary bypass grafting, and all recovered with nonoperative therapy. A total of 52 patients were followed up 1-52 months (24.5 months on average). All patients survived and the symptoms of basilar and upper l imb artery ischemia disappeared. Doppler ultrasonography showed that the blood flow was patent through anastomosis and polytetrafluoroethylene graft, and the vertebral artery flow was normal. Pseudoaneurysm at anastomosis was found in 1 case after 18 months and treated by interventional embol ization. The postoperative graft patency rate was 100% at 1 year and at 2 years. Conclusion Both thoracic and extrathoracic surgical approaches are effective for treating subclavianartery occlusion. The reasonable surgical approach should be selected according to the arteriopathy and the patient’s condition. Perioperative treatment and strict intraoperative manipulation are important to guarantee the success of surgery.