circ-Erbb2ip from adipose-derived mesenchymal stem cell-derived exosomes promotes wound healing in diabetic mice by inducing the miR-670-5p/Nrf1 axis.

BACKGROUND: To investigate the mechanism of exosomes (Exo) secretion by hypoxic pretreated adipose-derived mesenchymal stem cells (ADSCs) promoting skin wound healing in diabetic (DM) mice. METHODS: High-throughput sequencing was used to investigate abnormal expression of circRNA in hypoxic pretreatment ADSCs exosome (HExo) and ADSCs exosome (Exo). Bioinformatics analysis and luciferase reporting analysis were used to clarify the interacted relationship among circRNA, miRNA and mRNA. EPCs cells were employ to analysis the ROS, inflammatory cytokines expression, angiogenic differentiation function under hypoxic condition by using immunofluorescence, ELISA detection and tube forming experiment. DM ulceration mice model were constructed and the therapeutic effect of Exo were detected using immunohistochemistry, immunofluorescence. RESULTS: The result show that HExo have more treatment effect than Exo in promotes cutaneous wound healing of DM mice. High-throughput sequencing found that circ-Erbb2ip play a role in HExo mediated tissues repair. Downregulation circ-Erbb2ip decreased the therapeutic effect of HExo to wound healing in diabetic mice. Bioinformatics analysis and luciferase reporting analysis confirmed that both miR-670-5p and Nrf1 were downstream targets of circ-Erbb2ip. Downregulation of Nrf1 or overexpression of miR-670-5p reversed the protective effect of circ-Erbb2ip to EPCs after exposure to high glucose microenvironment. Upregulation circ-Erbb2ip increased the therapeutic effect of Exo to wound healing in diabetic mice by increased angiogenesis and decreased ROS, inflammatory cytokines expression. CONCLUSION: In conclusion, ADSC-Exos containing circ-Erbb2ip promotes wound healing by targeting miR-670-5p/Nrf1 pathway, and their effects in promoting soft tissue wound healing warrant further study.

Adipose-derived stem cell-secreted exosomes enhance angiogenesis by promoting macrophage M2 polarization in type 2 diabetic mice with limb ischemia via the JAK/STAT6 pathway.

Diabetic lower limb ischemia is an intractable disease that leads to amputation and even death. Recently, adipose-derived stem cell-secreted exosomes (ADSC-Exo) have been reported as a potential therapeutic approach, but its specific mechanism of action is unknown. Studies have found that exosomes derived from stem cells can reduce inflammation and promote tissue repair. Macrophages play an important role in the development and repair of inflammation in lower limb ischemic tissue, but the specific regulation of ADSC-Exo in macrophages has rarely been reported. The present study aimed to verify whether ADSC-Exo could promote angiogenesis by regulating macrophages to reduce the level of inflammation in diabetic ischemic lower limbs. In this study, adipose-derived stem cells (ADSCs) were obtained and identified, and ADSC-Exos were isolated using ultracentrifugation and characterized using transmission electron microscopy, nanoparticle tracking analysis, and western blotting analysis. The uptake of ADSC-Exos by macrophages was observed using immunofluorescence, and macrophage polarization induced by ADSC-Exos was identified by flow cytometry, immunofluorescence and ELISA. The effects of ADSC-Exos on the proliferation, apoptosis, migration and adhesion of macrophages were evaluated using CCK-8 assay, flow cytometry, Transwell assay, scratch and adhesion experiments, and ELISA assay. The polarization-related JAK/STAT6 signaling pathway was explored by using western blotting. A lower limb ischemic model of type 2 diabetic mice was established and ADSC-Exos was intramuscularly injected into the mice. The blood flow in the lower limbs was assessed using a laser Doppler flowmeter, while the level of angiogenesis was determined using immunohistochemistry and immunofluorescence. The results of this study prove that ADSC-Exos induced M2-phenotype polarization of macrophages via the JAK/STAT6 signaling pathway can promote the proliferation, migration and adhesion of M2 macrophages, inhibit the apoptosis of macrophages, and promote the angiogenesis and revascularization of ischemic lower limbs in type 2 diabetic mice. Thus, this study provides a theoretical and experimental basis for the clinical treatment of diabetic lower limb ischemic disease.

Study on the Features of Clinical Imaging Diagnosis of Arteriosclerotic Encephalopathy.

This paper discusses the imaging diagnostic features of arteriosclerotic encephalopathy and combines the spatial context information of local features to study the clinical imaging image copy detection algorithm. Moreover, this paper proposes a clinical imaging copy detection algorithm that combines the BOW model and spatial context embedding and a clinical imaging copy detection algorithm that combines the BOW model and global context verification. In addition, this paper applies the algorithm to the imaging diagnostic features of arteriosclerotic encephalopathy and sets up a controlled experiment to start research. The experimental research shows that the application of imaging diagnosis to the detection of subcortical arteriosclerotic encephalopathy has good clinical effects and rapid remission of patients' symptoms. The effectiveness of this method can be verified by a large number of clinical practices in follow-up studies.

Study on the Effect and Mechanism of miR-185 on Lower Extremity Deep Venous Thrombosis.

Lower extremity deep venous thrombosis (LEDVT) is a venous reflux disorder caused by abnormal coagulation of blood. LEDVT can obstruct the lumen and LEDVT is the third vascular disease after cerebrovascular diseases and coronary artery diseases. miRNAs are associated with thrombosis, and miR-185 was reported to affect the proliferation and apoptosis of vascular endothelial cells by regulating receptor of advanced glycation end products (RAGE). However, no study has reported the effect of miR-185 on LEDVT. Here, we studied the effects of miR-185 on the PI3K/AKT and MAPK signaling pathways in the LEDVT cells. The results showed that miR-185 promotes cell proliferation through activating the PI3K/AKT and MAPK signaling pathways and then inhibits tissue factor and fibrin expression to reduce thrombosis. In short, our study provides new ideas and a theoretical basis for research on the prevention, diagnosis, and treatment of LEDVT.

MicroRNA-125b inhibits the proliferation of vascular smooth muscle cells induced by platelet-derived growth factor BB.

Excessive proliferation and migration of vascular smooth muscle cells (VSMCs) is the main cause of arteriosclerosis obliterans (ASO). The present study aimed to investigate the role of microRNA (miR)-125b on the proliferation and migration of VSMCs. Platelet-derived growth factor-BB (PDGF-BB; 20 ng/ml) was used to treat VSMCs to establish an in vitro model of ASO. VSMCs were transfected with miR-125b mimic to overexpress miR-125. Cell Counting kit-8 (CCK-8) and BrdU assays were performed to assess the proliferative ability of VSMCs, while Transwell and wound healing assays were performed to assess the migratory ability of VSMCs. Western blot and immunofluorescence analyses were performed to detect the expression levels of angio-associated migratory cell protein (AAMP) and serum response factor (SRF) in VSMCs following transfection with miR-125b mimic or inhibitor. The results demonstrated that miR-125b expression decreased following treatment with PDGF-BB, the effects of which were reversed following transfection with miR-125b mimic. According to the CCK-8 assay, the cell proliferative ability decreased by ~50% compared with the negative control (NC) group, and ~40% at day 4 based on the BrdU assay. The results of the Transwell and wound healing assays indicated that the migratory ability of VSMCs significantly decreased in the miR-125b mimic group compared with the NC group. Furthermore, western blot and immunofluorescence analyses demonstrated that AAMP and SRF expression levels decreased following transfection with miR-125b mimic compared with the NC group, the effects of which were reversed following transfection with miR-125 inhibitor. Taken together, the results of the present study suggested that miR-125b inhibits the proliferative and migratory abilities of VSMCs by regulating the expression levels of AAMP and SRF.

Cu3BiS3 Nanocrystals as Efficient Nanoplatforms for CT Imaging Guided Photothermal Therapy of Arterial Inflammation.

Cardio-cerebrovascular diseases caused by chronic inflammatory atherosclerosis seriously damage human health. Nano-photothermal technology has been proven to inhibit the development of vascular inflammation, but the currently reported photothermal agents cannot efficient monitor it during the development of the disease. Herein, we designed and prepared an efficient bifunctional nanoplatform for CT imaging guided photothermal therapy of arterial inflammation. Cu3BiS3 nanocrystals with a size of about 12 nm were synthesized by a simple hydrothermal method. The as-prepared Cu3BiS3 nanocrystals showed intense absorption in the NIR region, thus exhibited amazing photothermal effect. The photothermal conversion efficiency of Cu3BiS3 nanocrystals was reach up to 58.6% under the excitation of an 808 nm laser with a power density of 0.4 W cm-2. Cu3BiS3 nanocrystals can efficiently kill the macrophages both in vitro and in vivo, which plays an important role in the development of atherosclerosis, thus can be used as an effective way to inhibit the occurrence of hypertension. Importantly, Cu3BiS3 nanocrystals can be used as an efficient CT contrast agent to monitor carotid inflammation. Our work provides an insight for imaging guided photothermal therapy of arterial inflammation.

MicroRNA-15b participates in the development of peripheral arterial disease by modulating the growth of vascular smooth muscle cells.

As an atherosclerotic disease, the process of peripheral arterial disease (PAD) is complicated and includes the abnormal proliferation of vascular smooth muscle. The current study aimed to determine the role of microRNA-15b (miR-15b) in the development of PAD and its associated mechanisms. Human vascular smooth muscle cells (hVSMCs) were used in the current study. To assess the effects of miR-15b on hVSMCs, miR-15b was up- or downregulated in hVSMCs using miR-15b mimics or miR-15b inhibitors respectively. Cell viability, migration and apoptosis were then determined via MTT, transwell and flow cytometry assays, respectively. TargetScan bioinformatics software was utilized to predict the targets of miR-15b, and the binding sites between insulin growth factor 1 receptor (IGF1R) and miR-15b were confirmed by dual-luciferase reporter assay. The results reveled that the miR-15b mimic significantly reduced hVSMC cell viability and migration, and promoted cell apoptosis. However, the opposite effect was observed following miR-15b inhibitor transfection. It was also determined that miR-15b directly targeted IGF1R and negatively regulated its expression in hVSMCs. Additionally, the results demonstrated that the miR-15b mimic inhibited the PI3K/AKT signaling pathway in hVSMCs, whereas the miR-15b inhibitor promoted it. Furthermore, the results indicated that the effect of the miR-15b mimic on hVSMCs was reversed by IGF1R overexpression. In conclusion, the data indicated that miR-15b participated in the occurrence and development of PAD by modulating hVSMC proliferation, apoptosis and migration via the regulation of IGF1R expression.

Effectiveness and Postoperative Prognosis of Using Preopening and Staged Percutaneous Transluminal Angioplasty of the Inferior Vena Cava in Treating Budd-Chiari Syndrome Accompanied with Inferior Vena Cava Thrombosis.

BACKGROUND: Budd-Chiari syndrome (BCS) is a rare disorder that has relatively high prevalence in the Huang-Huai area of China. Effective treatment of BCS accompanied with inferior vena cava thrombosis is challenging. OBJECTIVE: This study retrospectively analyzed the clinical effectiveness and safety of traditional open operations versus preopening and staged percutaneous transluminal angioplasty (PTA) of the inferior vena cava in treating BCS accompanied with inferior vena cava thrombosis. METHODS: Data from patients hospitalized and treated for BCS accompanied with inferior vena cava thrombosis between January 1997 and December 2017 were retrospectively analyzed. Thirty-two patients received traditional open operation (open group). Fifty-six patients received preopening and staged PTA of the inferior vena cava (PTA group). Baseline and clinical data were compared between groups. The patients were followed for up to 60 months. Postoperative recurrence rates and restenosis degree were recorded. RESULTS: Eighty-eight patients were included (47 males and 41 females), aged 41.82 ± 10.12 years (range 29-65). In the open group, no pulmonary arterial embolism was found during and after the operation, and the technique success rate was 100%. One patient died of intrathoracic bleeding. In the PTA group, 2 patients had shifting of thrombus in the inferior vena cava that blocked the blood flow restored by the preopening, one resulted in treatment failure, while the other had blood flow restored by dilation with a 12-mm balloon; the success rate was 55/56 (98.21%). The median follow-up time was 32 months (range 3-60). Two patients in the open group developed restenosis 2 years after operation (recurrence rate: 6.25%), and were successfully treated by balloon PTA. Seven patients in the PTA group had severe restenosis 18-42 months after operation (recurrence rate: 12.96%). No thrombosis was found in these 7 patients, and normal blood flow was restored in the inferior vena cava after balloon PTA. CONCLUSIONS: Preopening and staged PTA of the inferior vena cava is a safe and simple method for the treatment of BCS accompanied with inferior vena cava thrombosis, with satisfactory treatment effectiveness that could be applied in clinical practice.

A canine model for IVC occlusive form of Budd-Chiari syndrome using endovascular technique.

The objective of this study was to assess the portal/hepatic changes in a newly designed canine model for Budd-Chiari syndrome (BCS). The inferior vena cava (IVC) was occluded using a diagram stent under general anesthesia in 10 mongrel dogs under the guidance of percutaneous angiography. Five dogs that received IVC angiography only were used as sham controls. Occlusion of the IVC increased the diameter of the hepatic veins, portal vein, and IVC. Massive ascites, significantly increased abdominal circumference, varying degrees of esophageal varices, congestion, cirrhosis, and fibrosis of the liver were also noted. BCS could be readily established by placing a diaphragm-stent in the IVC via a percutaneous endovascular approach.

[Endovascular treatment of multilevel arteriosclerosis obliterans of lower extremities].

OBJECTIVE: To evaluate the clinical effect of percutaneous transluminal angioplasty (PTA) and PTA and stenting (PTAS) in treating multilevel arteriosclerosis obliterans (ASO) of lower extremities. METHODS: Between January 2007 and October 2009, 29 cases of multilevel ASO of lower extremities were treated with PTA or PTAS. There were 17 males and 12 females with an average age of 71 years (range, 56-83 years). The clinical manifestations were limbs numb with cold sensation, intermittent claudication (< 100 m), and 19 patients had rest pain. According to Fontaine stages classification, there were 7 cases of stage II, 14 cases of stage III, and 8 cases of stage IV. Concomitant diseases included hyperlipidemia in 13 cases, diabetes in 6 cases, hypertension in 12 cases, chronic heart failure in 5 cases, chronic obstructive pulmonary disease in 3 cases, cerebral infarction in 5 cases, and toe ulcer in 15 cases. The preoperative ankle/brachial index (ABI) was 0.28 +/- 0.13. RESULTS: In 3 cases of below knee lesions for the guide wire puncturing through the artery wall, 1 case gave up, and 2 cases continued recanalized other arteries without serious bleeding complications. After operation, cerebral infarction occurred in 1 case, pulmonary infection in 2 cases, and groin puncture site hematoma in 2 cases. All patients were followed up 3 to 36 months. Limb skin temperature was significantly improved, and intermittent claudication disappeared or claudication distance lengthened, rest pain disappeared or significantly reduced in 27 patients (30 limbs). The wound of amputee achieved good healing in 2 cases (2 limbs). In 27 patients undergoing endovascular treatment, the postoperative ABI was 0.72 +/- 0.19, showing significant difference when compared with the preoperative one (P < 0.05). CONCLUSION: The PTA and PTAS treatment for multilevel ASO of lower extremities is a safe and effective option, and the short-term effect is good. Different treatments should be chosen according to the location and length of vascular occlusion.

[Effect of urokinase on morphological changes of vein wall after acute thrombosis in rabbits].

OBJECTIVE: To assess the effect of urokinase on morphological changes of vein wall after acute thrombosis in rabbits. METHODS: Forty-three rabbits were randomly divided into 3 groups: thrombolysis group (n = 20), non-thrombolysis group (n = 20) and control group (n = 3). The thrombosed femoral veins of four rabbits in thrombolysis group anf four rabbits in non-thrombolysis group were taken 1, 4, 7, 14, and 21 days after thrombosis. The expression areas of smooth muscle actin and collagen fiber were measured by histochemistry. Scanning electron microscopy was used to observe the damage of endothelial cells. RESULTS: The expression area of smooth muscle actin began to increase significantly since day 4 after thrombosis reached the peak value on day 7 in thrombolysis group and non-thrombolysis group; and then decreased in thrombolysis group but remained at a high level in non-thrombolysis group. The expression area of smooth muscle actin in thrombolysis group was always less that in non-thrombosis group (all P < 0.01). The expression area of collagen fiber began to remarkably increase since day 7 after thrombosis in both thrombolysis and non-thrombolysis groups, however, being less in the thrombolysis group, and then decreased in thrombolysis group and remained at a high level in non-thrombosis group. Different cellular damages were observed by SEM one day after thrombosis in both thrombosis groups. However, the injury cells began to be repaired since day 4 in thrombolysis group. CONCLUSION: Thrombolysis with urokinase effectively maintains the integrity of endothelium of vein wall and decreases the fibrocellular proliferation of the thrombosed vein, however, fails to stop the pathological changes caused by thrombosis.

[Surgical treatment of left iliac compression syndrome].

OBJECTIVE: To explore the remedial indications, operational methods, and perioperational treatment of iliac compression syndrome (ICS) in left extremity. METHODS: The clinical data of 46 patients with symptoms of chronic swelling or varicose vein in left lower extremity and the diagnosis of left ICS were analyzed. Interventional treatment, iliac vein loosening by laparotomy and right iliac artery anastomosis behind left iliac vein lesion, and vascular graft orthotopic transplantation were performed in 32, 8 and 6 cases respectively. Surgical management of saphenous vein and femoral valves were performed simultaneously or by secondary operation in 41 cases. 36 patients were complicated by acute left lower extremity deep venous thrombosis (DVT), of which resection of lesion in left iliac vein and vascular graft orthotopic transplantation after thrombectomy, and iliocaval bypass were performed in 25 and 11 cases respectively. After treatment, the patients were followed up for 6 - 66 months. RESULTS: No operation death and acute pulmonary embolism was found among the patients. The treatment showed a good long-term effect. CONCLUSION: Lesions of left iliac vein are the important cause of swelling and varicose veins and acute DVT in left lower extremity and should be managed as early as possible. Interventional treatment and surgery are effective.