Carotid Injury Research Articles

CFTR Suppresses Neointimal Formation Through Attenuating Proliferation and Migration of Aortic Smooth Muscle Cells.

Cystic fibrosis transmembrane conductance regulator (CFTR) plays important roles in arterial functions and the fate of cells. To further understand its function in vascular remodeling, we examined whether CFTR directly regulates platelet-derived growth factor-BB (PDGF-BB)-stimulated vascular smooth muscle cells (VSMCs) proliferation and migration, as well as the balloon injury-induced neointimal formation. The CFTR adenoviral gene delivery was used to evaluate the effects of CFTR on neointimal formation in a rat model of carotid artery balloon injury. The roles of CFTR in PDGF-BB-stimulated VSMC proliferation and migration were detected by mitochondrial tetrazolium assay, wound healing assay, transwell chamber method, western blot, and qPCR. We found that CFTR expression was declined in injured rat carotid arteries, while adenoviral overexpression of CFTR in vivo attenuated neointimal formation in carotid arteries. CFTR overexpression inhibited PDGF-BB-induced VSMC proliferation and migration, whereas CFTR silencing caused the opposite results. Mechanistically, CFTR suppressed the phosphorylation of PDGF receptor β, serum and glucocorticoid-inducible kinase 1, JNK, p38 and ERK induced by PDGF-BB, and the increased mRNA expression of matrix metalloproteinase-9 and MMP2 induced by PDGF-BB. In conclusion, our results indicated that CFTR may attenuate neointimal formation by suppressing PDGF-BB-induced activation of serum and glucocorticoid-inducible kinase 1 and the JNK/p38/ERK signaling pathway.

Optimal Timing of Operative Intervention for High-grade Carotid Injuries in Patients with Blunt Trauma: A Nationwide Analysis of Trauma Quality Improvement Program

Blunt carotid injuries (BCIs) are rare injuries with an incidence of 1% to 3% in patients with blunt trauma. Earlier diagnosis and treatment are necessary to prevent the morbidity associated with BCIs. Operative intervention (open or endovascular) is indicated for patients with high-grade BCIs. However, there are no guidelines regarding the timing of operative intervention. The aim of our study was to assess the impact of early vs late operative intervention for BCIs.

Lipid phosphate phosphatase‐3 dysfunction exaggerates neointimal hyperplasia following diabetic vascular injury

IntroductionDiabetes mellitus is a major risk factor for cardiovascular diseases and promotes neointimal hyperplasia characterized by occlusive disorders such as atherosclerosis and stenosis, caused by excessive proliferation and migration of vascular smooth muscle cells (VSMCs) following endovascular interventions such as balloon angioplasty and stent placement. Lipid phosphate phosphatase‐3 (LPP3) is an enzyme that regulates the availability of lysophosphatidic acid (LPA) and has been reported to play a significant role in vascular genesis, cellular permeability, tumor progression, vascular remodeling, and myocardial infarction. However, the functional role and mechanism of LPP3 in diabetic neointimal hyperplasia is unclear. LPP3 dysfunction may lead to LPA accumulation at the vascular injury site, leading to vascular inflammation, proliferation, and migration, causing restenosis. Recently, the Receptor for Advanced Glycation End product (RAGE) has been presented as a potential receptor for LPA leading to downstream signaling in vascular smooth muscle cells (VSMCs).MethodsIn this study, we used a streptozocin‐induced diabetes mice model. We made SM‐22 specific LPP3 knock‐out mice and performed endothelial denudation surgery in the carotid artery to induce neointima hyperplasia to tease out the role of LPP3 in restenosis. We isolated aortic VSMCs from LPP3 floxed and LPP3 knock‐out mice and cultured them under normal or high glucose media. The immunoblotting was performed and probed with antibodies for total extracellular signal‐regulated kinase (ERK) and phosphorylated‐ERK. The cell proliferation and migration assay was performed using the Electric Cell substrate Impedance Sensing (ECIS) with or without LPP3 in VSMC under normal or high glucose conditions. Data are mean ± SEM; P‐values were determined by Student t‐test or one‐way ANOVA with Tukey's post‐hoc tests.ResultWe observed a significant increase in proliferation (P<0.01) and exaggerated migration (p<0.01) of VSMC that lack LPP3 under the hyperglycemic condition in real‐time using ECIS. These effects are at least partially a consequence of decreased lysophosphatidic acid hydrolysis due to lack of LPP3 activity. Furthermore, VSMC that lack LPP3 showed increased phospho‐ERK expression (P<0.01) under hyperglycemic conditions. The diabetic LPP3 knock‐out mice exhibited exaggerated neo‐intimal areas after the carotid injury compared to LPP3 floxed mice.ConclusionThese findings implicate that LPP3 plays a vital role as a negative regulator of smooth muscle phenotypic modulation. We speculate that changes in LPP3 expression control the development of neo‐intima by limiting smooth muscle cell proliferation, migration, and vascular inflammation in diabetic restenosis.

α-Lipoic Acid-Plus Ameliorates Endothelial Injury by Inhibiting the Apoptosis Pathway Mediated by Intralysosomal Cathepsins in an In Vivo and In Vitro Endothelial Injury Model.

α-Lipoic acid-plus (LAP), an amine derivative of α-lipoic acid, has been reported to protect cells from oxidative stress damage by reacting with lysosomal iron and is more powerful than desferrioxamine (DFO). However, the role of LAP in experimental carotid artery intimal injury (CAII) has not yet been well investigated. Therefore, we sought to uncover the role and potential endovascular protective mechanisms of LAP in endothelial injury. In vitro, oxyhemoglobin (OxyHb) stimulation of cultured human umbilical vein endothelial cells (HUVECs) simulated intimal injury. In vivo, balloon compression injury of the carotid artery was used to establish a rat CAII model. We found that the protein levels of cathepsin B/D, ferritin, transferrin receptor (TfR), cleaved caspase-3, and Bax increased in the injured endothelium and HUVECs but were rectified by DFO and LAP treatments, as revealed by western blotting and immunofluorescence staining. Additionally, DFO and LAP decreased oxidative stress levels and endothelial cell necrosis of the damaged endothelium. Moreover, DFO and LAP significantly ameliorated the increased oxidative stress, iron level, and lactic dehydrogenase activity of HUVECs and improved the reduced HUVEC viability induced by OxyHb. More importantly, DFO and LAP significantly reduced mitochondrial damage and were beneficial for maintaining lysosomal integrity, as indicated by acridine orange (AO), Lyso-Tracker Red, JC-1, and ATPB staining in HUVECs. Finally, LAP might offer more significant endovascular protective effects than DFO. Our data suggested that LAP exerted endovascular protective effects by inhibiting the apoptosis signaling pathway mediated by intralysosomal cathepsins by reacting with excessive iron in endothelial lysosomes after intimal injury.

Platelet-derived microvesicles regulate vascular smooth muscle cell energy metabolism via PRKAA after intimal injury

Vascular intimal injury initiates various cardiovascular disease processes. Exposure to subendothelial collagen can cause platelet activation, leading to collagen-activated platelet-derived microvesicles (aPMVs) secretion. In addition, vascular smooth muscle cells (VSMCs) exposed to large amounts of aPMVs undergo abnormal energy metabolism; they proliferate excessively and migrate after the loss of endothelium, eventually contributing to neointimal hyperplasia. However, the roles of aPMVs in VSMC energy metabolism are still unknown. Our carotid artery intimal injury model indicated that platelets adhered to injured blood vessels. In vitro, phosphorylated Pka (cAMP-dependent protein kinase) content was increased in aPMVs. We also found that aPMVs significantly reduced VSMC glycolysis and increased oxidative phosphorylation, and promoted VSMC migration and proliferation by upregulating phosphorylated PRKAA (α catalytic subunit of AMP-activated protein kinase) and phosphorylated FoxO1. Compound C, an inhibitor of PRKAA, effectively reversed the enhancement of cellular function and energy metabolism triggered by aPMVs in vitro and neointimal formation in vivo. We show that aPMVs can affect VSMC energy metabolism through the Pka-PRKAA-FoxO1 signaling pathway and this ultimately affects VSMC function, indicating that the shift in VSMC metabolic phenotype by aPMVs can be considered a potential target for the inhibition of hyperplasia. This provides a new perspective for regulating the abnormal activity of VSMCs after injury.

Code-free machine learning for object detection in surgical video: a benchmarking, feasibility, and cost study

While the utilization of machine learning (ML) for data analysis typically requires significant technical expertise, novel platforms can deploy ML methods without requiring the user to have any coding experience (termed AutoML). The potential for these methods to be applied to neurosurgical video and surgical data science is unknown. AutoML, a code-free ML (CFML) system, was used to identify surgical instruments contained within each frame of endoscopic, endonasal intraoperative video obtained from a previously validated internal carotid injury training exercise performed on a high-fidelity cadaver model. Instrument-detection performances using CFML were compared with two state-of-the-art ML models built using the Python coding language on the same intraoperative video data set. The CFML system successfully ingested surgical video without the use of any code. A total of 31,443 images were used to develop this model; 27,223 images were uploaded for training, 2292 images for validation, and 1928 images for testing. The mean average precision on the test set across all instruments was 0.708. The CFML model outperformed two standard object detection networks, RetinaNet and YOLOv3, which had mean average precisions of 0.669 and 0.527, respectively, in analyzing the same data set. Significant advantages to the CFML system included ease of use, relatively low cost, displays of true/false positives and negatives in a user-friendly interface, and the ability to deploy models for further analysis with ease. Significant drawbacks of the CFML model included an inability to view the structure of the trained model, an inability to update the ML model once trained with new examples, and the inability for robust downstream analysis of model performance and error modes. This first report describes the baseline performance of CFML in an object detection task using a publicly available surgical video data set as a test bed. Compared with standard, code-based object detection networks, CFML exceeded performance standards. This finding is encouraging for surgeon-scientists seeking to perform object detection tasks to answer clinical questions, perform quality improvement, and develop novel research ideas. The limited interpretability and customization of CFML models remain ongoing challenges. With the further development of code-free platforms, CFML will become increasingly important across biomedical research. Using CFML, surgeons without significant coding experience can perform exploratory ML analyses rapidly and efficiently.

A Comorbid Rat Model of Neuroendocrine-Immune System Alterations Under the Impact of Risk Factors for Stroke.

Hypercholesterolemia and carotid atherosclerosis contribute to the etiology of stroke. However, there has been a lack of appropriate comorbid animal models incorporating some of the ubiquitous characteristics that precede strokes. Curcumin is a natural active polyphenolic compound extracted from the rhizoma of Curcuma longa L. which possesses comprehensive bioactivities. The present study aimed to evaluate whether neurobehavioral deficits, neuroendocrine-immune dysregulations and cerebral microcirculation dysfunction, are part of the initial stages of cerebral ischemia in individuals suffering from carotid atherosclerosis resulting from a high cholesterol diet (HCD) and if they could be tested using a comorbid animal model. Furthermore, the utility of this model will be examined following the administration of curcumin. Adult wild-type SD rats were fed a regular diet or HCD and supplemented with either vehicle or curcumin for 4 weeks. Carotid injury was induced by an air-drying endothelial denudation method at the end of the second week. Plasma cholesterol, carotid pathomorphology, neurobehavioral tests, and neuroendocrine-immune parameters were measured. We found higher plasma levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein-cholesterol (LDL-C), intima and media (I/M) ratio, but lower high-density lipoprotein-cholesterol (HDL-C), spatial learning and memory capacity impairment, elevated NPY expression in the hypothalamus, increased plasma concentration of leptin, upregulated TNF-α, IL-1β, and CRP in the circulation as well as TNF-α and IL-1β in the cerebral cortex, plus enhanced ICAM-1, VCAM-1, and E-selectin in cerebral microvessels in HCD-fed model rats. All these alterations were ameliorated by curcumin. These results suggest that a comorbid rat model was effectively developed by HCD and carotid injury.

GDF11 alleviates neointimal hyperplasia in a rat model of artery injury by regulating endothelial NLRP3 inflammasome activation and rapid re-endothelialization

BackgroundNeointimal hyperplasia induced by interventional surgery can lead to progressive obliteration of the vascular lumen, which has become a major factor affecting prognosis. The rate of re-endothelialization is known to be inversely related to neointima formation. Growth differentiation factor 11 (GDF11) is a secreted protein with anti-inflammatory, antioxidant, and antiaging properties. Recent reports have indicated that GDF11 can improve vascular remodeling by maintaining the differentiated phenotypes of vascular smooth muscle cells. However, it is not known whether and how GDF11 promotes re-endothelialization in vascular injury. The present study was performed to clarify the influence of GDF11 on re-endothelialization after vascular injury.MethodsAn adult Sprague–Dawley rat model of common carotid artery balloon dilatation injury was surgically established. A recombinant adenovirus carrying GDF11 was delivered into the common carotid artery to overexpress GDF11. Vascular re-endothelialization and neointima formation were assessed in harvested carotid arteries through histomolecular analysis. CCK-8 analysis, LDH release and Western blotting were performed to investigate the effects of GDF11 on endothelial NLRP3 inflammasome activation and relevant signaling pathways in vitro.ResultsGDF11 significantly enhanced re-endothelialization and reduced neointima formation in rats with balloon-dilatation injury by suppressing the activation of the NLRP3 inflammasome. Administration of an endoplasmic reticulum stress (ER stress) inhibitor, 4PBA, attenuated endothelial NLRP3 inflammasome activation induced by lysophosphatidylcholine. In addition, upregulation of LOX-1 expression involved elevated ER stress and could result in endothelial NLRP3 inflammasome activation. Moreover, GDF11 significantly inhibited NLRP3 inflammasome-mediated endothelial cell pyroptosis by negatively regulating LOX-1-dependent ER stress.ConclusionsWe conclude that GDF11 improves re-endothelialization and can attenuate vascular remodeling by reducing endothelial NLRP3 inflammasome activation. These findings shed light on new treatment strategies to promote re-endothelialization based on GDF11 as a future target.

The hidden aspect of carotid lesions in traffic accidents: A little-known phenomenon

Vascular injuries that occur during traffic accidents are a commonly neglected aspect that can add more detail to the framework of a case. In this study, we analysed a case series of 150 traffic accidents, 39 of which were marked by microscopically identifiable vascular lesions. The purpose was to identify the presence of carotid injuries in individuals who died due to traffic accidents and had nonpenetrating trauma of the neck. We focused on the discrepancies regarding the macroscopical aspect and the histology and demonstrated how histological analysis of the carotids in cases of trauma can reveal injuries that are attributable to the trauma itself. We conducted a histological analysis of the lesions to describe their distribution and type and investigate potential correlations. The study offers insight on how to examine road accidents that involve traumatic injury of the carotid arteries. Indeed the main task of the forensic pathologist in the case of death is to establish the existence of a causal relationship between the micro- or macroscopic alterations observed in the autopsy and the traumatic event that led to the death of the subject. Thus, further morphological elements were provided to the forensic practitioners that may reveal injuries attributable to the trauma itself and should be evaluated in cases of trauma in traffic accidents.

Risk factors for stroke in penetrating carotid trauma-An analysis from the PROOVIT Registry.

Penetrating carotid injuries are associated with an up to 20% risk of stroke. This study evaluated patients in the American Association for Surgery of Trauma PROspective Observational Vascular Injury Trial, with the aim of determining factors associated with stroke and stroke or death. Penetrating extracranial carotid injuries in the American Association for Surgery of Trauma PROspective Observational Vascular Injury Trial registry from 2012 to 2020 were queried. Isolated external carotid injuries were excluded. Patients with documented postinjury in-hospital stroke were compared with those without. Significant predictors (p < 0.1) for stroke and stroke or death on univariate analysis were included in multivariate analyses. One hundred two patients from 17 institutions were included. Mean age was 35 ± 18 years, and 80% were male. Average Glasgow Coma Scale (GCS) score on presentation was 9 ± 5, with an Injury Severity Score [ISS] of 22 ± 13. Operative management occurred in 51% of patients who were significantly more hypotensive (systolic blood pressure: 109 vs. 131 mm Hg; p = 0.015) with a lower initial pH (7.17 vs. 7.31; p = 0.001) and presented with hard signs of vascular injury (74% vs. 26%; p < 0.001). Overall stroke rate was 17% (23% operative vs. 10% nonoperative, p = 0.076). Rate of stroke or death was 27% (64% operative and 36% nonoperative). On multivariate analysis, lower GCS (p = 0.05) and completion angiography (p = 0.04) were associated with stroke. Likewise lower GCS (p = 0.015) and ISS (p = 0.04) were associated with stroke or death. Penetrating carotid trauma undergoing operative management had a stroke rate of 23%. Low GCS on arrival and need for completion angiography are independently associated with postinjury in-hospital stroke, whereas low GCS on arrival and ISS were associated with stroke or death. The ideal treatment strategy remains elusive, thus a dedicated multicenter study may help to achieve higher fidelity data on this rare but devastating injury. Prognostic and Epidemiological, Level III.

Recombinant Hirudin Protects the Vasculature and Myocardium in Atherosclerotic Rats by Inhibiting the p38 MAPK/NF-kB Signaling Pathway

Recombinant hirudin (r-hirudin) has a good anticoagulant effect and has a certain inhibitory effect on atherosclerosis (AS), however, its intrinsic mechanism of inhibiting AS is still unclear. In this study, we investigated the mechanism underlying the vascular and myocardial protective effects of r-hirudin in AS rats through animal experiments. A rat AS model was established by high-fat diet feeding combined with common carotid artery balloon injury. The model rats were given low, medium, or high doses of r-hirudin (0.05, 0.1, or 0.2 mg/kg/ day), simvastatin tablets (1 mg/kg/day) and p38 mitogen-activated protein kinase (MAPK) pathway inhibitors (SB203580, 100 mg/kg/day) by gavage for 8 weeks. The results showed that in AS rats, r-hirudin significantly alleviated pathological changes in the common carotid artery and myocardial tissue; decreased serum total cholesterol (TC), triglyceride (TG) and low-density lipoprotein-cholesterol (LDL-C) levels; increased highdensity lipoprotein-cholesterol (HDL-C) levels; decreased serum oxidized low-density lipoprotein (ox-LDL), tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β, and endothelin (ET)-1 levels; increased nitric oxide (NO) levels; and decreased p38 MAPK, nuclear factor-kappa B (NF-κB), caspase-9, caspase-3 mRNA and protein expression. This study showed that r-hirudin may protect blood vessels and the myocardium in AS rats by adjusting blood lipid levels and inhibiting the p38 MAPK/NF-κB signaling pathway to exert anti-inflammatory and anti-apoptotic effects and protect the vascular endothelium.

Galangin inhibits neointima formation induced by vascular injury via regulating the PI3K/AKT/mTOR pathway.

Aims: The proliferation and migration of vascular smooth muscle cells (VSMCs) play vital roles in the pathological process of neointima formation after vascular injury. Galangin, an extract of the ginger plant galangal, is involved in numerous biological activities, including inhibiting the proliferation and migration of tumor cells, but its effect on VSMCs is unknown. This study focused on the role and mechanism of galangin in the neointima formation induced by vascular injury. Methods and results: In this study, we found that galangin restrained the PDGF-BB-induced proliferation, migration and phenotypic switching of VSMCs in a concentration-dependent manner. In vivo, we established a model of carotid artery balloon injury in rats, followed by intragastric administration of galangin (40 mg kg-1 day-1 or 80 mg kg-1 day-1) for 14 or 28 consecutive days. Then, the degree of neointima hyperplasia was evaluated by H&E staining, and the level of relevant protein expression was assessed by immunofluorescence and western blotting. In vitro, we isolated and grew primary rat aortic smooth muscle cells, which were treated with PDGF-BB and different doses of galangin, and then CCK-8 assay, wound healing assay, transwell assay, western blotting and immunofluorescence assays were performed. We found that galangin significantly inhibited PDGF-BB-induced proliferation, migration, and phenotypic switching of VSMCs and promoted autophagy in VSMCs in vitro, and galangin significantly inhibited neointimal hyperplasia after the common carotid artery balloon injury in rats. In terms of mechanisms, galangin inhibited the PI3K/AKT/mTOR pathway, thereby suppressing VSMC's switch from a contractile to a synthetic phenotype, inhibiting VSMC proliferation, migration and phenotypic switching and upregulating the Beclin1 protein expression levels and the ratio of LC3BII/I, promoting VSMC autophagy, and thereby inhibiting neointimal hyperplasia after vascular injury. Conclusion: Our study suggests that galangin inhibits neointimal hyperplasia after vascular injury by inhibiting smooth muscle cell proliferation, migration and phenotypic switching and by promoting autophagy, and that galangin may be a promising drug for the prevention and treatment of vascular restenosis after PCI.

Internal Carotid Injury during Transsphenoidal Resection of Pituitary Macroadenoma Managed with Neurointervention Technique

AbstractIatrogenic internal carotid artery (ICA) injuries during transsphenoidal pituitary tumor surgery are extremely rare but are associated with potentially disabling or life-threatening complications. Management options of ICA injuries are sacrifice, embolization, and endoluminal reconstruction. We report a case of ICA injury during transsphenoidal pituitary macroadenoma surgery, which was managed with endovascular covered stent placement for left carotid-ophthalmic pseudoaneurysm.

Case report; successful treatment of traumatic ischaemic hemiplegia secondary to blunt carotid injury associating high grade liver trauma

Introduction and importanceBlunt carotid injury (BCI) injury is a rare sequel of trauma and could result in ischemic complication if not detected and treated early. The presence of high-grade solid organ injury with ongoing bleeding represents additional challenge in treating BCI.Case presentationA 25-year-old victim of motor vehicle collision resulted in grade IV liver, grade III left kidney and grade I spleen injury. He underwent an urgent laparotomy with transient liver packing at local hospital. A full body Contrast-Enhanced Computer Tomography (CECT) upon arrival revealed right internal carotid intimal tear with intra and extra-cranial thrombosis and a 3 cm aneurysm. With a decreased level of consciousness, the patient showed a GCS of 13 and left-sided hemiplegia. After complex multidisciplinary treatment sessions, patient recovered with a partial regain of left-sided muscle power.Clinical discussionSelective embolization of active liver bleeding was a turning point in the management of our patient as it deferred the need for a second operative intervention. It was a necessary step before endovascular stenting and recanalization of the BCI to restore the circulation to the right cerebral hemisphere. Dual anti-platelet therapy (DAPT) was necessary to prevent thrombosis of the stent and continuity of carotid recanalization.ConclusionBCI with traumatic ischaemic hemiplegia associating a sum of life-threatening multiple injuries including high grade liver trauma with ongoing bleeding could still be managed non-operatively with acceptable outcome in the presence of a comprehensive specialized multidisciplinary service.

Sex Differences in Arterial Identity Are Associated with Neointimal Hyperplasia after Carotid Balloon Injury

Sex Differences in Arterial Identity Are Associated with Neointimal Hyperplasia after Carotid Balloon Injury

JCAD enhances arterial thrombosis by regulating endothelial plasminogen activator inhibitor-1 and tissue factor expression

Abstract Introduction Arterial thrombosis underlies most acute CV events. Variants of the Junctional cadherin 5 associated (JCAD) locus were consistently shown to associate with increased risk of acute coronary syndrome. Being a component of cell junctions, JCAD protein is highly expressed in endothelial cells and was shown to promote atherosclerosis by acting on the Hippo pathway through LATS2 kinase. Purpose This project investigated the effect of JCAD in arterial thrombosis by using an established in vivo mouse model of carotid injury. The translational value of animal findings was assessed in primary human aortic endothelial cells (HAECs) as well as in CV patients. Methods JCAD knock-out (Jcad−/−) mice were exposed to photochemically-induced carotid artery endothelial injury to trigger thrombosis. Primary HAECs treated with JCAD small-interfering RNA (si-JCAD), LATS2-silencing RNA (si-LATS2) or control siRNA (si-SCR) were employed for in vitro assays. Plasma JCAD was measured in patients with chronic coronary syndrome (CCS) or ST-elevation myocardial infarction (STEMI). Results Compared to wild-type, Jcad−/− mice displayed reduced thrombus formation as underlined by delayed time to occlusion following endothelial-specific carotid damage. Suggesting a blunted activation of the extrinsic coagulation cascade, Jcad−/− animals showed reduced tissue factor (TF) protein expression and activity in carotid artery lysates (Fig. 1). Increased thrombus embolization episodes and D-dimer further suggested an increased activation of the fibrinolytic system in Jcad−/− mice. Indeed, Jcad−/− mice displayed reduced vascular expression of the fibrinolysis inhibitor plasminogen activator inhibitor (PAI)-1. In contrast, platelets aggregation in response to collagen and thrombin was similar in Jcad−/− and Jcad+/+ mice (Fig. 1). In line with the in vivo data, JCAD-silencing of HAECs inhibited TF and PAI-1 gene and protein expression. In accordance with previous literature, JCAD-silenced HAECs displayed increased levels of LATS2 Kinase, which blunts the Hippo pathway by increasing YAP phosphorylation. Yet, double JCAD and LATS2 silencing did not retrieve the phenotype of control HAECs. Of interest, si-JCAD HAECs showed increased levels of Akt phosphorylation, known to downregulate procoagulant expression and to directly phosphorylate YAP. Treatment with the Akt inhibitor Wortmannin prevented the effect of JCAD silencing on TF and PAI-1 indicating a causative role for this pathway (Fig. 2). Recapitulating in vitro findings, p-Akt and p-YAP levels were higher in arterial tissue of Jcad−/− animals as compared to WT (Fig. 1). Patients with STEMI showed significantly higher plasma levels of JCAD as compared to CCS (Fig. 2). Conclusions JCAD promotes arterial thrombosis by selectively modulating coagulation and fibrinolysis, but not platelet aggregation through endothelial TF and PAI-1. Our findings support the importance of JCAD as a novel therapeutic target for CV prevention. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Swiss National Science Foundation

675 ICA Ligation and Malignant MCA Syndrome Secondary to A Gunshot Wound: A Case Presentation

Abstract Penetrating carotid injuries occur infrequently in the UK. Without surgical intervention, mortality rates are close to 100%. Whilst vessel repair is the optimal surgical choice, zone III neck injuries or haemodynamic instability in particular often require vessel ligation, which carries higher mortality rates, increased risk of stroke and poorer outcomes overall. We present a young patient who sustained a gunshot wound to the neck. Emergency exploration revealed a penetrating injury to zone III of the left side of the neck and a complete transection of the left internal carotid artery. Due to haemodynamic instability, damage control surgery was mandated, and the internal carotid artery was ligated. The patient was admitted to ITU but had a severe acute neurological deterioration post-operation. CT head imaging revealed a large left frontal-parietal infarct with rightward midline shift, in keeping with malignant MCA syndrome. He underwent an emergency decompressive craniectomy, with resolution of the midline shift seen on subsequent imaging. At 3 months, the patient walked independently out of hospital, with a reduction in his modified Rankin Scale score from 5 to 2 following intensive therapies input. Whilst rare, the importance of timely recognition and intervention of an infrequent complication of carotid ligation is paramount. Decompressive craniectomy is the mainstay of treatment for malignant MCA syndrome, yet outcomes remain poor in all ages, highlighting the patient’s noteworthy recovery. Though emerging evidence suggests novel endovascular repair techniques may provide favourable outcomes and reduced complications in carotid injuries, open surgery remains the treatment of choice.

Carotid Artery-Cavernous Segment Injury during an Endoscopic Endonasal Surgery: A Case Report and Literature Review of the Overlooked Option for Surgical Trapping in the Hyperacute Phase.

Internal carotid artery (ICA) injury is a catastrophic complication of endoscopic endonasal surgery (EES). However, its standard management, emergent endovascular treatment, may not always be available, and the transnasal approach may be insufficient to achieve hemostasis.A 44-year-old woman with pituitary adenoma underwent EES complicated with the ICA cavernous segment injury (CSI). In urgent intraoperative angiogram, a good collateral flow from the contralateral carotid circulation was observed. Due to the unavailability of intraoperative embolization, emergent surgical trapping was performed by combined transcranial and cervical approach. The patient recovered but later developed a giant cavernous pseudoaneurysm. During the pseudoaneurysm embolization, ICA was directly accessed via a 1.7-F puncture hole using a bare microcatheter technique. Then, both the aneurysm and parent artery were obliterated with coils. At the 4-year follow-up, the patient was asymptomatic without a residual tumor. To our knowledge, this is the first case of ICA–CSI during EES successfully treated with ICA trapping as a lifesaving urgent surgery that achieved a complete recovery after a pseudoaneurysm embolization. Although several studies reported that EES-related ICA–CSIs with percutaneous carotid artery access, neither our surgical salvage technique nor our carotid access and tract embolization techniques were previously described.

A new Mfn-2 related synthetic peptide promotes vascular smooth muscle cell apoptosis via regulating the mitochondrial apoptotic pathway by inhibiting Akt signaling

BackgroundRestenosis after angioplasty is a major challenge for the treatment of coronary artery diseases. Facilitation of vascular smooth muscle cell (VSMC) apoptosis may be an attractive approach to decrease the incidence of restenosis. We synthesized a 16-amino acid mitofusin-2 (Mfn-2) gene related peptide (MRSP) based on the sequence of the p21ras signature motif, the smallest functional sequence of the Mfn-2 gene with proapoptotic properties in VSMC. We investigated whether MRSP enhanced apoptotic activities to inhibit VSMC accumulation and neointimal hyperplasia in rats with carotid balloon injury.MethodsVSMCs were treated with different concentrations of MRSP, the PI3K agonist 740 Y-P and the inhibitor LY294002. Cell apoptosis and related pathway molecules were assessed. MRSP was also given to rats with carotid artery balloon injury. Neointimal hyperplasia and cell apoptotic pathways were detected.ResultsIn vitro experiments revealed that MRSP treatment significantly increased VSMC apoptosis and induced increases in procaspase-9 cleavage, caspase-3 activation, cytochrome c release from mitochondria to the cytoplasm and the Bax/Bcl-2 ratio but not caspase-8 expression, indicating that the mitochondrial apoptotic cascade was activated by MRSP, which might be attributed to suppression of the PI3K/Akt signaling pathway. We further found that the PI3K agonist 740 Y-P prevented and that the inhibitor LY294002 strengthened the proapoptotic effects of MRSP. MRSP strongly inhibited neointimal hyperplasia and VSMC accumulation, but increased VSMC apoptosis in the vascular wall after balloon injury. Moreover, MRSP substantially enhanced Bax and cleaved caspase-3 expression and decreased Bcl-2 levels in intima, accompanied by decreased levels of phosphorylated Akt and PI3K in vivo.ConclusionsTaken together, the present study showed that MRSP treatment results in a strong proapoptotic effect by activating the mitochondrial apoptotic cascade through suppression of the PI3K/Akt pathway.

Human urinary kininogenase reduces the endothelial injury by inhibiting Pyk2/MCU pathway

The injury of endothelial cells is one of the initiating factors in restenosis after endovascular treatment. Human urinary kallidinogenase (HUK) is a tissue kallikrein which is used for ischemia-reperfusion injury treatment. Studies have shown that HUK may be a potential therapeutic agent to prevent stenosis after vascular injury, however, the precise mechanisms have not been fully established. This study is to investigate whether HUK can protect endothelial cells after balloon injury or H2O2-induced endothelial cell damage through the proline-rich tyrosine kinase 2 (Pyk2)/mitochondrial calcium uniporter (MCU) pathway. Intimal hyperplasia, a decrease of pinocytotic vesicles and cell apoptosis were found in the common carotid artery balloon injury and H2O2-induced endothelial cell damage, Pyk2/MCU was also up-regulated in such pathological process. HUK could prevent these injuries partially via the bradykinin B2 receptor by inhibiting Pyk2/MCU pathway, which prevented the mitochondrial damage, maintained calcium balance, and eventually inhibited cell apoptosis. Furthermore, MCU expression was not markedly increased if Pyk2 was suppressed by shRNA technique in the H2O2 treatment group, and cell viability was significantly better than H2O2-treated only. In short, our results indicate that the Pyk2/MCU pathway is involved in endothelial injury induced by balloon injury or H2O2-induced endothelial cell damage. HUK plays an protective role by inhibiting the Pyk2/MCU pathway in the endothelial injury.