TRIM64 promotes ox-LDL-induced foam cell formation, pyroptosis, and inflammation in THP-1-derived macrophages by activating a feedback loop with NF-κB via IκBα ubiquitination.

Atherosclerosis is a chronic inflammatory disease and the main pathology behind most cardiovascular diseases and the overactivation of macrophages initiates the development of atherosclerosis. However, the specific functions of oxidized low-density lipoprotein (ox-LDL) in macrophages remain elusive. Macrophages derived from monocyte (THP-1) were treated with ox-LDL and were used to generate atherosclerosis in an in vitro model. NLRP3 inflammasome markers were examined using quantitative RT-PCR and Western blotting. Cytokines were measured using ELISA. Chromatin immunoprecipitation (ChIP) was utilized to detect nuclear factor kappa B (NF-κB) and TRIM64 interactions. A fat-rich diet was applied to ApoE-/- mice for in vivo studies. ox-LDL promoted TRIM64 expression in a time-dependent manner. According to loss- and gain-of-function analyses, TRIM64 enhanced the activation of NLRP3 inflammasomes and the expression of downstream molecules. TRIM64 directly interacted with IκBα and promoted IκBα ubiquitination at K67 to activate NF-κB signaling. We detected direct binding between NF-κB and the TRIM64 promoter, as well as enhanced TRIM64 expression. Our study revealed an interaction between TRIM64 and NF-κB in the development of atherosclerosis. TRIM64 and NF-κB formed a positive feedback to activate NF-κB pathway. ox-LDL induces foam cell formation and TRIM64 expression TRIM64 regulates ox-LDL-induced foam cell formation, pyroptosis and inflammation via the NF-κB signaling TRIM64 activates NF-κB signaling by ubiquitination of IκBα NF-κB inhibition attenuates atherosclerosis in HFD-induced ApoE (-/-) mice.

Delineating the conformational landscape and intrinsic properties of the angiotensin II type 2 receptor using a computational study.

As a key regulator for the renin-angiotensin system, a class A G protein-coupled receptor (GPCR), AngII type 2 receptor (AT2R), plays a pivotal role in the homeostasis of the cardiovascular system. Compared with other GPCRs, AT2R has a unique antagonist-bound conformation and its mechanism is still an enigma. Here, we applied combined dynamic and evolutional approaches to investigate the conformational space and intrinsic properties of AT2R. With molecular dynamic simulations, Markov State Models, and statistics coupled analysis, we captured the conformational landscape of AT2R and identified its uniquity from both dynamical and evolutional viewpoints. A cryptic pocket was also discovered in the intermediate state during conformation transitions. These findings offer a deeper understanding of the AT2R mechanism at an atomic level and provide hints for the design of novel AT2R modulators.

Acute myocardial infarction as the first sign of infective endocarditis: a case report.

Infective endocarditis is a bacterial or fungal infection of the heart valves or endocardial surface, and it frequently forms vegetation and can lead to systemic embolism. Dislodged vegetation rarely results in coronary artery embolism (CAE) and subsequent acute myocardial infarction. A 43-year-old male patient was emergently brought to our hospital for suspected acute myocardial infarction. Coronary angiography was performed and it showed embolism in the left circumflex artery. Thrombus aspiration was performed during coronary angiography. Echocardiography showed formation of vegetation in the posterior leaflet of the mitral valve and multiple blood cultures showed Listeria monocytogenes. Infective endocarditis was diagnosed. Three weeks later, debridement of subacute bacterial endocarditis, mitral valve replacement, and tricuspid valvuloplasty were successfully conducted. Our findings suggest that CAE should be considered in the differential diagnosis of acute myocardial infarction. Aspiration of coronary embolus during coronary angiography followed by surgical intervention of diseased heart valves is a plausible strategy for managing CAE in infective endocarditis.

Circadian locomotor output cycles kaput accelerates atherosclerotic plaque formation by upregulating plasminogen activator inhibitor-1 expression.

To explore the association between clock circadian regulator circadian locomotor output cycles kaput gene (CLOCK) and the forming of atherosclerotic plaques and its underlying mechanisms, mouse aortic endothelial cells (MAECs) and atherosclerosis (AS) mouse model were recruited for our study. The apoE gene knockout mouse was used as the model of AS and we accelerated the formation of unstable plaques through the combination of carotid artery ligation and high-fat (HF) diet administration (0.2% cholesterol, 20% fat). The mRNA and protein expressions of CLOCK in peripheral blood monouclear cells of acute coronary syndrome (ACS) patients or mouse AS model were detected by qPCR, western blot analysis and immunohistochemical staining. The number of adherent cells and atherosclerotic plaques was counted to assess the effects of CLOCK on the progression of ACS, and adherence-associated genes, such as vascular cell adhesion molecule (VCAM)-1, C-C motif chemokine ligand 2 (CCL-2), and CCL-5. The results showed that CLOCK expression was significantly increased in both ACS patients and AS mouse model. The levels of CLOCK, leukemia inhibitory factor (LIF), intercellular adhesion molecule 1 (ICAM-1), perilipin 2 (ADFP), nuclear factor kappa B (NF-κB), and plasminogen activator inhibitor-1 (PAI-1), as well as the number of atherosclerotic plaques were elevated in the AS mouse model, as compared with the control group. Chromatin immunoprecipitation assay showed that CLOCK bound directly to the promoter of PAI-1 gene and CLOCK could positively regulate the expressions of LIF, ICAM-1, ADFP, NF-κB, and PAI-1. Reduction of CLOCK expression would decrease the expressions of VCAM-1, CCL-2, and CCL-5, and the number of adherent cells and atherosclerotic plaques, but these effects were neutralized when PAI-1 was simultaneously overexpressed in either mouse model or MAECs. Our results demonstrate that CLOCK overexpression triggers the formation of atherosclerotic plaques by directly upregulating PAI-1 expression.

PGC-1α ameliorates AngiotensinII-induced eNOS dysfunction in human aortic endothelial cells.

Increasing evidences support that PGC-1α participates in regulating endothelial homeostasis, in part by mediating endothelial nitric oxide (NO) synthase (eNOS) activity and NO production. However, the molecular mechanisms by which PGC-1α regulates eNOS activity are not completely understood. In the present study, we investigated the effects of PGC-1α on eNOS dysfunction and further explore the underlying mechanisms. The results showed that PGC-1α expression was downregulated after AngiotensinII (AngII) treatment and paralleled with the decreased NO generation in human aortic endothelial cells. Overexpression of PGC-1α with adenovirus or pharmacological agonist ameliorated AngII-induced the decrease of NO generation, evidenced by the restoration of cGMP and nitrite concentration. Rather than affecting eNOS expression and uncoupling, PGC-1α inhibited AngII-induced decrease of eNOS serine 1177 phosphorylation through activation of PI3K/Akt signaling. In addition, PGC-1α overexpression suppressed AngII-induced the increase of PP2A-A/eNOS interaction and PP2A phosphatase activity, with a concomitant decrease in PP2A phosphorylation, leading to eNOS serine 1177 phosphorylation. However, pharmacological inhibition of PI3K/Akt signaling blunted the observed effect of PGC-1α on PP2A activity. Taken together, our findings suggest that PGC-1α overexpression improves AngII-induced eNOS dysfunction and that improved eNOS dysfunction is associated with activated PI3K/Akt pathway, impaired PP2A activity and reduced PP2A-A/eNOS association. These date indicate that forced PGC-1α expression may be a novel therapeutic approach for endothelial dysfunction.

Randomized Comparison of 3 Hemostasis Techniques After Transradial Coronary Intervention.

BACKGROUND: The transradial route for coronary intervention has proven to be a safe and feasible method, and several techniques have been shown to be effective in achieving hemostasis. OBJECTIVES: The aim of this study is to evaluate the efficacy of 3 hemostasis techniques on radial artery outcomes after transradial catheterization. METHODS: A total of 1650 patients were randomly assigned to 1 of the 3 hemostasis techniques after the procedure. The outcome measures were local vascular complications, tolerance to the device, and the time taken to achieve hemostasis. RESULTS: Time taken to achieve hemostasis was significantly longer in the pressure dressing (PD) group than in the pneumatic compression device (PCD) group and rotary compression pad device (RCD) group (306 ± 65 vs 263 ± 62 and 237 ± 58 minutes; P < .0001). There were also significant differences between PD, PCD, and RCD groups with respect to the incidence of oozing (8.2% vs 5.1% and 5.1%; P = .047) and discomfort level (1.68 vs 1.43 and 1.40; P < .0001). The incidence of early (24 hours after the procedure) radial artery occlusion was significantly higher in the PD group than in the PCD and RCD groups (15.6% vs 5.8% and 4.5%; P < .0001). Logistic regression analysis showed that independent predictors of radial artery occlusion at 30-day follow-up visit were diabetes (hazard ratio, 2.39), larger radial artery diameter (hazard ratio, 0.52), the use of the PCD (hazard ratio, 0.51, compared with PD) and the RCD (hazard ratio, 0.52, compared with PD), and radial artery patency during compression (hazard ratio, 0.016). CONCLUSIONS: Hemostasis devices have comparative advantages over the conventional pressure dressing. The presence of radial artery flow during compression represents a strong predictor of radial artery patency during follow-up periods.

IL-18 upregulates the production of key regulators of osteoclastogenesis from fibroblast-like synoviocytes in rheumatoid arthritis.

Recent data have demonstrated the importance of IL-18 in the induction and perpetuation of chronic inflammation in experimental arthritis. The aim of the present study was to elucidate whether IL-18 has any indirect effects on osteoclastogenesis by regulating the production of molecules from fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA). Human FLS were isolated from RA synovial tissue and cultured in vitro for three to five passages. The expression of IL-18 receptor was determined by RT-PCR. The levels of soluble receptor activator of nuclear factor κB ligand (RANKL), osteoprotegerin (OPG), macrophage colony-stimulating factor (M-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) in culture supernatants were determined by ELISA. Membrane-bound RANKL expression was analyzed by flow cytometry. Both α and ß chains of IL-18 receptor were confirmed in cultured FLS. IL-18 upregulated membrane-bound RANKL expression and soluble RANKL production by FLS in both time- and dose-dependent manners. In addition, IL-18 enhanced production of M-CSF, GM-CSF, and OPG from cultured FLS in a dose-dependent manner. IL-18 also increased the ratio of RANKL/OPG, suggesting that the net effect of IL-18 on FLS favors for the induction of osteoclast formation and bone resorption. In conclusion, IL-18 upregulates the production of key regulators of osteoclastogenesis from FLS in RA.

Takayasu's arteritis: clinical features and outcomes of 125 patients in China.

Takayasu's arteritis (TA) is a chronic inflammation that frequently involves the aorta and its major branches. The clinical features of TA vary in different ethnic populations. The objective of this study is to characterize the clinical features, angiographic findings, and response to treatment of patients with TA in Changhai Hospital, Shanghai, China. The hospital records of 125 patients diagnosed with TA were retrospectively evaluated. Eighty patients were followed for a median duration of 36 months. Females (86.4%) were most frequently affected. The mean age at onset was 26.9 years. Constitutional symptoms were present in only 38.4% of patients. The most common clinical finding was pulse deficit. Histological findings from 12 clinically inactive patients showed active lesions in 58.3%. Angiographic classification showed that type I was the most common, followed by type V and IV. Type I was more common in adult patients than in pediatric patients. Although immunosuppressive treatment induced remission in most patients, over 90% of those who achieved later remission relapsed. Both bypass procedures and angioplasty showed high rates of initial success, but restenosis occurred in 34.7% of bypass procedures and 77.3% of angioplasty procedures. Eight patients died during the follow-up period with the main cause of death being congestive heart failure. Constitutional symptoms were not frequent in our study. Correlation between the clinical assessment of disease activity and histologic findings is often poor in TA. Angiographic findings showed that type I was the most common in our study. Over the longer term, the outcomes of revascularization were superior to angioplasty.

The use of calcium sulfate impregnated with vancomycin in the treatment of open fractures of long bones: a preliminary study.

The purpose of this study was to investigate the effect of calcium sulfate impregnated with vancomycin combined with internal fixation in the treatment of open fractures of long bones. Between October 2007 and January 2008, twenty-eight patients (24 men, 4 women) who sustained open fractures of long bones were enrolled. Mean patient age was 34.5 years (range, 19-57 years). According to the classification system developed by Gustilo and Anderson, there were 15 type II open fractures, 11 type IIIa open fractures, and 2 type IIIc open fractures. All the patients were treated with vancomycin-loaded calcium sulphate and internal fixation. Clinical signs of wound infection were recorded. Postoperative radiographs were used to evaluate the bone healing and absorption process.Twenty-six of 28 patients were followed-up successfully, with an average follow-up of 10.5 months (range, 6-16 months). No infection was present in the 26 patients. Exudation from the incision or the drain incision was observed in 2 patients. Bacteria culture was negative and the wound healed with dressing change within 15 days. Bone union was observed in 23 patients, with a mean bone union time of 5.8 months (range, 4-9 months). All the calcium sulfate pellets had completely resorbed in an average of 1.4 months (range, 1-2 months). Our study showed that the combination of internal fixation and calcium sulfate impregnated with vancomycin could decrease the incidence of deep infection without impairment to the bone healing process in the treatment of open fractures of long bones.