MiR-127-3p enhances macrophagic proliferation via disturbing fatty acid profiles and oxidative phosphorylation in atherosclerosis.

BACKGROUND: Atherosclerosis is a chronic pathology, leading to acute coronary heart disease or stroke. MiR-127 has been found significantly upregulated in advanced atherosclerosis. But its function in atherosclerosis remains unexplored. We explored the role of miR-127-3p in regulating atherosclerosis development and its downstream mechanisms. METHODS: The expression profile of miR-127 in carotid atherosclerotic plaques of 23 patients with severe carotid stenosis was detected by RT-qPCR and in situ hybridization. Primary bone marrow-derived macrophages (BMDM) stimulated with oxidized low-density lipoprotein were used as an in vitro model. CCK-8, EdU, RT-qPCR, and flow cytometry were used to detect the proliferative capacity and polarization of BMDM, which were infected by lentivirus-carrying plasmid to upregulate or downregulate miR-127-3p expression, respectively. RNA sequencing combined with bioinformatic analysis and targeted fatty acid metabolomics approach were used to detect the transcriptome and lipid metabolites. The association between miR-127-3p and its target was verified by dual-luciferase activity reporting and Western blotting. Oxygen consumption rate of BMDM were detected using seahorse analysis. High-cholesterol-diet-fed low density lipoprotein deficient (LDLR-/-) mice, with-or-without carotid tandem-stenosis surgery, were treated with miR-127-3p agomir or antagomir to examine its effect on plaque development and stability. RESULTS: miR-127-3p, not -5p, is elevated in human advanced carotid atheroma and its expression is positively associated with macrophage accummulation in plaques. In vitro, miR-127-3p-overexpressed macrophage exhibites increased proliferation capacity and facilitates M1 polariztion whereas the contrary trend is present in miR-127-3p-inhibited macrophage. Stearoyl-CoA desaturase-1 (SCD1) is one potential target of miR-127-3p. miR-127-3p mimics decreases the activity of 3' untranslated regions of SCD-1. Furthermore, miR-127-3p downregulates SCD1 expression, and reversing the expression of SCD1 attenuates the increased proliferation induced by miR-127-3p overexpression in macrophage. miR-127-3p overexpression could also lead to decreased content of unsaturated fatty acids (UFAs), increased content of acetyl CoA and increased level of oxidative phosphorylation. In vivo, miR-127-3p agomir significantly increases atherosclerosis progression, macrophage proliferation and decreases SCD1 expression and the content of UFAs in aortic plaques of LDLR-/- mice. Conversely, miR-127-3p antagomir attenuated atherosclerosis, macrophage proliferation in LDLR-/- mice, and enhanced carotid plaque stability in mice with vulnerable plaque induced. CONCLUSION: MiR-127-3p enhances proliferation in macrophages through downregulating SCD-1 expression and decreasing the content of unsaturated fatty acid, thereby promoting atherosclerosis development and decreasing plaque stability. miR-127-3p/SCD1/UFAs might provide potential therapeutic target for anti-inflammation and atherosclerosis.

METTL3-mediated m6A modification of NORAD inhibits the ferroptosis of vascular smooth muscle cells to attenuate the aortic dissection progression in an YTHDF2-dependent manner.

Ferroptosis of vascular smooth muscle cells (VSMCs) is related to the incidence of aortic dissection (AD). Long non-coding RNA (lncRNA) NORAD plays a crucial role in the progression of various diseases. The present study aimed to investigate the effects of NORAD on the ferroptosis of VSMCs and the molecular mechanisms. The expression of NORAD, HUR, and GPX4 was detected using quantitative real-time PCR (qPCR) or western blot. Ferroptosis was evaluated by detecting lactate dehydrogenase (LDH) activity, lipid reactive oxygen species (ROS), malonaldehyde (MDA) content, L-Glutathione (GSH) level, Fe2+ content, and ferroptosis-related protein levels. The molecular mechanism was assessed using RNA pull-down, RNA-binding protein immunoprecipitation (RIP), and luciferase reporter assay. The histology of aortic tissues was assessed using H&E, elastic Verhoeff-Van Gieson (EVG), and Masson staining assays. The data indicated that NORAD was downregulated in patients with AD and AngII-treated VSMCs. Overexpression of NORAD promoted VSMC growth and inhibited the ferroptosis induced by AngII. Mechanistically, NORAD interacted with HUR, which promoted GPX4 mRNA stability and elevated GPX4 levels. Knockdown of GPX4 abrogated the effects of NORAD on cell growth and ferroptosis of AngII-treated VSMCs. Moreover, METTL3 promoted m6A methylation of NORAD in an YTHDF2-dependent manner. In addition, NORAD attenuated AAD symptoms, incidence, histopathology, inflammation, and ferroptosis in AAD mice. In conclusion, METTL3-mediated NORAD inhibited ferroptosis of VSMCs via the HUR/GPX4 axis and decelerated AAD progression, suggesting that NORAD may be an AD therapeutic target.

Sorting Nexin 10 Mediates Metabolic Reprogramming of Macrophages in Atherosclerosis Through the Lyn-Dependent TFEB Signaling Pathway.

RATIONALE: SNX10 (sorting nexin 10) has been reported to play a critical role in regulating macrophage function and lipid metabolism. OBJECTIVE: To investigate the precise role of SNX10 in atherosclerotic diseases and the underlying mechanisms. METHODS AND RESULTS: SNX10 expression was compared between human healthy vessels and carotid atherosclerotic plaques. Myeloid cell-specific SNX10 knockdown mice were crossed onto the APOE-/- (apolipoprotein E) background and atherogenesis (high-cholesterol diet-induced) was monitored for 16 weeks. We found that SNX10 expression was increased in atherosclerotic lesions of aortic specimens from humans and APOE-/- mice. Myeloid cell-specific SNX10 deficiency (Δ knockout [KO]) attenuated atherosclerosis progression in APOE-/- mice. The population of anti-inflammatory monocytes/macrophages was increased in the peripheral blood and atherosclerotic lesions of ΔKO mice. In vitro experiments showed that SNX10 deficiency-inhibited foam cell formation through interrupting the internalization of CD36, which requires the interaction of SNX10 and Lyn-AKT (protein kinase B). The reduced Lyn-AKT activation by SNX10 deficiency promoted the nuclear translocation of TFEB (transcription factor EB), thereby enhanced lysosomal biogenesis and LAL (lysosomal acid lipase) activity, resulting in an increase of free fatty acids to fuel mitochondrial fatty acid oxidation. This further promoted the reprogramming of macrophages and shifted toward the anti-inflammatory phenotype. CONCLUSIONS: Our data demonstrate for the first time that SNX10 plays a crucial role in diet-induced atherogenesis via the previously unknown link between the Lyn-Akt-TFEB signaling pathway and macrophage reprogramming, suggest that SNX10 may be a potentially promising therapeutic target for atherosclerosis treatment.

Activation of Bone Marrow-Derived Cells and Resident Aortic Cells During Aortic Injury.

BACKGROUND: The process of aortic injury, repair, and remodeling during aortic aneurysm and dissection is poorly understood. We examined the activation of bone marrow (BM)-derived and resident aortic cells in response to aortic injury in a mouse model of sporadic aortic aneurysm and dissection. MATERIALS AND METHODS: Wild-type C57BL/6 mice were transplanted with green fluorescent protein (GFP)+ BM cells. For 4 wk, these mice were either unchallenged with chow diet and saline infusion or challenged with high-fat diet and angiotensin II infusion. We then examined the aortic recruitment of GFP+ BM-derived cells, growth factor production, and the differentiation potential of GFP+ BM-derived and GFP- resident aortic cells. RESULTS: Aortic challenge induced recruitment of GFP+ BM cells and activation of GFP- resident aortic cells, both of which produced growth factors. Although BM cells and resident aortic cells equally contributed to the fibroblast populations, we did not detect the differentiation of BM cells into smooth muscle cells. Interestingly, aortic macrophages were both of BM-derived (45%) and of non-BM-derived (55%) origin. We also observed a significant increase in stem cell antigen-1 (Sca-1)+ stem/progenitor cells and neural/glial antigen 2 (NG2+) cells in the aortic wall of challenged mice. Although some of the Sca-1+ cells and NG2+ cells were BM derived, most of these cells were resident aortic cells. Sca-1+ cells produced growth factors and differentiated into fibroblasts and NG2+ cells. CONCLUSIONS: BM-derived and resident aortic cells are activated in response to aortic injury and contribute to aortic inflammation, repair, and remodeling by producing growth factors and differentiating into fibroblasts and inflammatory cells.

Matrix metalloproteinases are regulated by MicroRNA 320 in macrophages and are associated with aortic dissection.

Aortic dissection (AD) is the circumferential or transversal tear of the aorta wall that allows blood to infiltrate the layers. MicroRNA (miR) analyses have demonstrated a correlation between miR-320 family and AD. The underlying mechanism is yet unclear. The matrix metalloproteinases (MMPs) are a group of proteolytic enzymes that could catalyze the degeneration of the extracellular matrix and the destruction of the vasculature. In this study, we investigated whether miR-320 presented a role in regulating the production of MMPs in aortic dissection. In a cohort of 30 CE patients and 30 healthy controls, the transcription and secretion of MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, and MMP-12 by monocytes were investigated. The monocyte from AD patients presented significantly elevated capacity of MMP expression than those from healthy controls. In contrast, the monocyte/macrophage expression of miR-320 was significantly lower in AD patients than in controls. In both AD patients and healthy controls, LPS-activation of macrophages resulted in MMP upregulation and miR-320 downregulation, in which the MMP expression was significantly higher while the miR-320 expression was significantly lower in AD patients than in healthy controls. Transfection of miR-320 mimic did not affect MMP gene transcription but significantly reduced the protein production in some MMPs, demonstrated that miR-320 were involved in the post-transcriptional regulation of MMPs. Together, these results demonstrated that miR-320 could regulate the expression of MMPs by macrophages, through which miR-320 may interfere with AD development.

Endovascular Therapy of a Giant Renal Artery Aneurysm Combined With High-Flow Renal Arteriovenous Fistula Using a Patent Ductus Arteriosus Occluder.

Renal artery aneurysm (RAA) concomitant with a renal arteriovenous fistula (RAVF) is extremely rare. A 32-year-old man suffered from a giant RAA combined with high-flow RAVF. The computer tomographic angiography (CTA) demonstrated a RAA, which is 6.3 cm in length and 2.1 cm in diameter, combined with an arteriovenous fistula between the right renal artery and right renal vein (fistula area:1.05 cm × 1.0 cm). After a comprehensive preoperative assessment, a patent ductus arteriosus occluder (PDAO) was implanted. At a 1-year follow-up, the CTA study showed that the PDAO was in situ and there was no recanalization of the lesion. At a third-year follow-up, ultrasound examination showed an image of right renal atrophy. The results of long-term follow-up demonstrate that PDAO is safe and effective for the management of RAAs combined with high-flow RAVF.

Heat shock protein 27 plays a protective role in thoracic aortic dissection by promoting cell proliferation and inhibiting apoptosis.

BACKGROUND: Thoracic aortic dissection (TAD) is one of the most severe aortic diseases. The study aimed to explore the potential role of heat shock protein 27 (HSP27) in the pathogenesis of TAD using an in vitro model of oxidative stress in vascular smooth muscle cells (VSMCs). METHODS: HSP27 was analyzed in aortic surgical specimens from 12 patients with TAD and 8 healthy controls. A lentiviral vector was used to overexpress HSP27 in rat aortic VSMCs. Cell proliferation and apoptosis were measured under oxidative stress induced by H2O2. RESULTS: HSP27 expression was significantly higher in aortic tissue from patients with TAD and VSMCs in the aortic media were the main cell type producing HSP27. Elevated oxidative stress was also detected in the TAD samples. Overexpression of HSP27 significantly attenuated H2O2-induced inhibition of cell proliferation. Furthermore, HSP27 was found to decrease H2O2-induced cell apoptosis and oxidative stress. CONCLUSIONS: These results suggest that HSP27 expression promotes VSMC viability, suppresses cell apoptosis, and confers protection against oxidative stress in TAD.

Improved visual, acoustic, and neurocognitive functions after carotid endarterectomy in patients with minor stroke from severe carotid stenosis.

OBJECTIVE: Carotid endarterectomy (CEA) is an established operation performed to prevent strokes, but its other potential effects, such as improving neurocognitive, visual, and auditory functions, remain unconfirmed. This study examined these effects of CEA on patients with symptomatic carotid stenosis. METHODS: This was a prospective controlled study that included 80 patients with minor strokes who had severe extracranial internal carotid stenoses (>70%). Forty patients, who did not receive or who postponed the CEA due to concerns about age, fear of surgery, limited life expectancy because of cancer, or financial problems, formed the medicine-treatment group. Another 40 patients who received CEA 1 week after recruitment formed the CEA group. For both groups, visual acuity chart tests, perimetry tests, audiometry tests, and neurologic scales (National Institutes of Health Stroke Scale, Mini Mental State Examination, and Barthel Index of Activities of Daily Living) were used to assess ophthalmic functions, auditory acuity, and neurocognitive functions before treatment and 3 months after treatment. Intragroup and intergroup comparisons were conducted to examine the effect of CEA. RESULTS: No deaths or strokes occurred during the 3-month follow-up. The intragroup and intergroup comparisons of ipsilateral function showed that CEA could improve visual acuity, visual field, and auditory acuity at all tested frequencies (250 Hz, 500 Hz, 1000 Hz, 2000 Hz, and 4000 Hz) and could improve the visual field and the auditory acuity for contralateral functions at 1000 Hz. The auditory acuity at 2000 Hz and 4000 Hz were unchanged in the intragroup comparison but showed no deterioration in the intergroup comparison with the medicine group. General neurocognitive function and independent living ability were significantly improved by CEA, as shown by intergroup comparisons (change rate of National Institutes of Health Stroke Scale: -8.1% ± 9.0% vs -2.7% ± 3.0%, P < .001; change rate of Mini Mental State Examination: 15.5% ± 10.5% vs 1.6% ± 2.6%, P < .001; change rate of Barthel Index: 28.0% ± 24.6% vs 2.0% ± 5.5%, P < .001). CONCLUSIONS: In patients with minor strokes caused by severe carotid stenosis, CEA improves neurocognitive, ophthalmic, and acoustic functions. Studies with a larger sample and longer follow-up are needed to substantiate these results, and the underlying mechanisms need further investigation.

AKT2 confers protection against aortic aneurysms and dissections.

RATIONALE: Aortic aneurysm and dissection (AAD) are major diseases of the adult aorta caused by progressive medial degeneration of the aortic wall. Although the overproduction of destructive factors promotes tissue damage and disease progression, the role of protective pathways is unknown. OBJECTIVE: In this study, we examined the role of AKT2 in protecting the aorta from developing AAD. METHODS AND RESULTS: AKT2 and phospho-AKT levels were significantly downregulated in human thoracic AAD tissues, especially within the degenerative medial layer. Akt2-deficient mice showed abnormal elastic fibers and reduced medial thickness in the aortic wall. When challenged with angiotensin II, these mice developed aortic aneurysm, dissection, and rupture with features similar to those in humans, in both thoracic and abdominal segments. Aortas from Akt2-deficient mice displayed profound tissue destruction, apoptotic cell death, and inflammatory cell infiltration that were not observed in aortas from wild-type mice. In addition, angiotensin II-infused Akt2-deficient mice showed significantly elevated expression of matrix metalloproteinase-9 (MMP-9) and reduced expression of tissue inhibitor of metalloproteinase-1 (TIMP-1). In cultured human aortic vascular smooth muscle cells, AKT2 inhibited the expression of MMP-9 and stimulated the expression of TIMP-1 by preventing the binding of transcription factor forkhead box protein O1 to the MMP-9 and TIMP-1 promoters. CONCLUSIONS: Impaired AKT2 signaling may contribute to increased susceptibility to the development of AAD. Our findings provide evidence of a mechanism that underlies the protective effects of AKT2 on the aortic wall and that may serve as a therapeutic target in the prevention of AAD.

[Surgical management of post carotid artery stenting restenosis].

OBJECTIVE: To discuss the feasibility, safety and effectiveness of surgical management of post carotid artery stenting (CAS) restenosis, mainly focusing on the surgical options and indications. METHODS: This study represented retrospective analysis of 3 kinds of surgical managements of 21 patients with symptomatic post CAS restenosis from April 2012 to April 2014. Patch carotid endarterectomy (pCEA), Eversion carotid endarterectomy (eCEA) or carotid excision and graft interposition (CEGI) was selected to remove the stent and reconstruct the blood flow, based on the preoperative imaging results and intraoperative adhesion degree. Use of carotid shunt, blood loss, operative time, carotid artery cross-clamp time and other data were recorded. Patients were followed for improvement of symptoms, complications and restenosis. RESULTS: Eleven, 4 and 6 patients received pCEA, eCEA or CEGI respectively. All the stents were successfully removed. Shunts were deployed in 14 cases. The mean bleeding was (152.6 ± 38.0) ml, the mean operation time was (100.7 ± 34.8) min and the mean carotid artery clamping time was (29.1 ± 4.6) min. In the early postoperative period, there were no infection, strokes, cranial nerve injury, myocardial infarction or mortalities. One patient developed neck hematoma, while 2 patients had the symptoms of hyperperfusion such as headache, irritability and multi-lingual but no intracranial hemorrhage happened according to the brain CT scan, who all fully recovered within 3 days. Within a median follow-up of (13.2 ± 4.3) months, no strokes, myocardial infarctions or recurrent restenosis (> 50%) on duplex ultrasound imaging or CTA was discovered except for 1 patient who died of lung cancer. CONCLUSION: Surgical management to remove the stent and reconstruct the blood flow, which offered new options in the treatment of post CAS restenosis, with its initially confirmed simplicity, feasibility, safety and validity.

[Single-center experience on clinical application of "anatomic fixation" unibody bifurcated stent-graft in endovascular repair].

OBJECTIVE: To evaluate the clinical outcomes of unibody bifurcated stent-graft in endovascular repair (EVAR). METHODS: Retrospective analyses were conducted for the clinical data and postoperative follow-up results of 125 patients (102 cases from Shanghai Changhai Hospital and another 23 from Shanghai Changzheng Hospital) undergoing EVAR with unibody bifurcated stent-graft from January 2008 to January 2013. RESULTS: The technical success rate was 100%. Perioperative complications including type I endoleak (n = 3, 2.4%) and type II endoleak (n = 4, 3.2%). The incidence of type I endoleak in challenging neck cases was higher than non-challenging ones. And the difference was statistically significant (P = 0.001). Except for 1 dead case, the remainder was followed up for a mean of 26.4 ± 1.5 (1-60) months. Neither aneurysm rupture nor stent-graft migration occurred. Late type I endoleak occurred (n = 2, 1.6%). There were left lower extremity arterial thrombosis (n = 1, 0.8%) and surgical reintervention (n = 1, 0.8%). Among 3 dead cases, 2 died from acute myocardial infarction and another 1 contrast-induced nephropathy. CONCLUSION: Unibody bifurcated stent-graft is both safe and efficacious in the treatment of abdominal aortic aneurysm without the risk of long-term migration. Moreover, it has excellent outcomes for hostile neck or narrow abdominal aortic bifurcation.

Comparative effectiveness and safety of laser, needle, and "quick fenestrater" in in situ fenestration during thoracic endovascular aortic repair.

Background: Special instruments are needed for the revascularization of aortic branches in in situ fenestration during thoracic endovascular aortic repair (TEVAR). This prospective study compared the effectiveness and safety of three currently used fenestraters: laser, needle, and Quick Fenestrater (QF). Methods: In all, 101 patients who underwent TEVAR for aortic disease (dissection, n = 62; aneurysm, n = 16, or ulcer, n = 23) were enrolled. All patients were randomly assigned to three groups: 34 were assigned to laser fenestration, 36 to needle fenestration, and 31 to QF fenestration. The epidemiological data, treatment, imaging findings, and follow-up outcomes were analyzed using data from the medical records. Results: The technical success rates of the laser, needle, and QF fenestration groups were 94.1%, 94.4%, and 100% (p > 0.05). After correction of mixed factors such as age and gender, it was showed the average operative time (Laser group: 130.01 ± 9.36 min/ Needle group: 149.80 ± 10.18 min vs. QF group: 101.10 ± 6.75 min, p < 0.001), fluoroscopy time (Laser group: 30.16 ± 9.81 min/ Needle group: 40.20 ± 9.91 min vs. QF group: 19.91 ± 5.42 min, p < 0.001), fenestration time (Laser group 5.50 ± 3.10 min / Needle group 3.50 ± 1.50 min vs. QF group 0.67 ± 0.06 min, p < 0.001), and guide wire passage time after fenestration (Laser group 5.10 ± 1.70 min / Needle group 4.28 ± 1.60 min vs. QF group 0.07 ± 0.01 min, p < 0.001) were all shorter with QF fenestration than with the other two tools. The overall perioperative complication rates of the laser, needle, and QF fenestration groups were 5.9%, 5.6%, and 0% (p > 0.05): One case of sheath thermal injury and one case of vertebral artery ischemia occurred in the laser fenestration group; one case each of access site hematoma and brachial artery thrombosis were reported in the needle fenestration group. 89 (88.1%, 89/101) patients were followed for a median of 12.6 ± 1.6 months. The overall postoperative complication rates of the laser, needle, and QF fenestration groups were 3.3%, 6.5%, and 0% (p > 0.05): In the laser fenestration group, there was one death due to postoperative ST-segment elevation myocardial infarction; in the needle fenestration group, one patient developed occlusion of the bridge stent; no complications occurred in the QF group. Conclusion: All three fenestration methods were effective in reconstructing supra-arch artery during TEVAR. QF fenestration required less contrast agent, with a shorter surgery duration and fewer complications than laser and needle fenestration.

Epitranscriptome analysis of NAD-capped RNA by spike-in-based normalization and prediction of chronological age.

Nicotinamide adenine dinucleotide (NAD) can be used as an initiating nucleotide in RNA transcription to produce NAD-capped RNA (NAD-RNA). RNA modification by NAD that links metabolite with expressed transcript is a poorly studied epitranscriptomic modification. Current NAD-RNA profiling methods involve multi-steps of chemo-enzymatic labeling and affinity-based enrichment, thus presenting a critical analytical challenge to remove unwanted variations, particularly batch effects. Here, we propose a computational framework, enONE, to remove unwanted variations. We demonstrate that designed spike-in RNA, together with modular normalization procedures and evaluation metrics, can mitigate technical noise, empowering quantitative and comparative assessment of NAD-RNA across different datasets. Using enONE and a human aging cohort, we reveal age-associated features of NAD-capping and further develop an accurate RNA-based aging clock that combines signatures from both transcriptome and NAD-modified epitranscriptome. enONE facilitates the discovery of NAD-RNA responsive to physiological changes, laying an important foundation for functional investigations into this modification.

Dengzhan Shengmai capsule versus Aspirin in the treatment of carotid atherosclerotic plaque: A single-centre, non-inferiority, prospective, randomised controlled trial.

BACKGROUND: Aspirin is an effective antiplatelet agent for the treatment of carotid atherosclerosis. However, the high risk of bleeding events associated with the drug makes it necessary to seek a safer alternative, with similar or more efficacy than aspirin. Dengzhan Shengmai (DZSM) capsules have been widely used to treat carotid atherosclerosis, and if proven to be non-inferior to aspirin, it may be preferable over the latter for carotid atherosclerosis treatment due to its numerous advantages. We conducted a randomised trial to test the non-inferiority of DZSM to aspirin for the treatment of carotid atherosclerotic plaques. METHODS: We performed a single-centre, prospective, open-label, randomised non-inferiority trial. Patients with carotid atherosclerotic plaques were enrolled and randomly assigned (1:1) to receive either DZSM capsules or aspirin. The follow-up period was 12 months. The primary outcome was the mean change in carotid intima-media thickness (IMT). Secondary outcomes included ischaemic events, rate of lumen stenosis, lipid levels, and plaque scores, length, counts, and vulnerability. Adverse events and laboratory test results were recorded as safety outcomes. The non-inferiority of DZSM was demonstrated when the lower limit of the one-sided 97.5% confidence interval (CI) of the difference in IMT between groups was more than -0.06 mm (margin of non-inferiority). This trial has been registered at ClinicalTrials.gov (CHiCTR1900021365). RESULTS: From 1 April 2019 to 30 September 2019, 150 patients were enrolled, and there was no statistical difference in demographics between the groups. Intention-to-treat analysis showed that the decrease in IMT(∆IMT) was 0.216 ± 0.160 and 0.225 ± 0.149 mm in the DZSM and aspirin groups, respectively. The one-sided 97.5% CI for the difference between ∆IMTs was (-0.0593, +∞). The non-inferiority of DZSM was demonstrated (Pnon-inferiority = 0.0234). There was no significant difference in the incidence of ischaemic events between the groups (P = 1.0). The DZSM group had significantly reduced plaque scores (P < 0.0001), length (P < 0.0001), and counts (P < 0.0001), and improved plaque vulnerability (P < 0.0001). The DZSM group also had reduced levels of low-density lipoprotein cholesterol (LDL-C) (P < 0.0001). Finally, the DZSM group had a lower incidence of total adverse events (14.7% vs. 28%, P = 0.046), especially gastrointestinal discomfort (5.3% vs. 16%, P = 0.034). Although there was no significant difference in bleeding events (0 vs. 5.3%, P = 0.120), the DZSM group tended to have a lower incidence. CONCLUSION: This trial demonstrated that DZSM was not inferior, in efficacy, to aspirin in treating carotid atherosclerotic plaques, and was found to be superior to aspirin in terms of safety. This study provides a new approach for treating carotid plaques, especially in aspirin-intolerant patients.

A microRNA profile comparison between thoracic aortic dissection and normal thoracic aorta indicates the potential role of microRNAs in contributing to thoracic aortic dissection pathogenesis.

OBJECTIVES: Our aim was to identify important microRNAs (miRNAs) that might play an important role in contributing to aortic dissection by conducting a miRNA profile comparison between thoracic aortic dissection (TAD) and normal thoracic aorta. METHODS: The differentially expressed miRNA profiles of the aortic tissue between TAD patients (n = 6) and age-matched donors without aortic diseases (NA; n = 6) were analyzed by miRNA microarray. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was further performed to verify the expression of 12 selected miRNAs with an increased number of samples (TAD n = 12; NA n = 8). The potential targets of the differentially expressed miRNAs were predicted using computational searches. Bioinformatic analyses of the predicted target genes (gene ontology, pathway and network analysis) were done for further research. Additionally, Western blotting was performed to confirm the bioinformatics findings. RESULTS: The miRNA microarray revealed differentially expressed miRNAs between the TAD and NA groups. In the TAD group, 18 miRNAs were upregulated and 56 were downregulated (fold change >2, P < .01). qRT-PCR verified statistically consistent expression of seven selected miRNAs with microarray analysis. Combined with our previous proteomics study, target gene prediction revealed that some miRNAs reciprocally expressed with their targeted proteins. Target gene-related pathway analysis showed a significant change in five pathways in the TAD group compared with the NA group, especially the focal adhesion and the mitogen-activated protein kinase (MAPK) signaling pathways. By further conducting miRNA gene network analysis, we found that the mir-29 and mir-30 families are likely to play a role in the regulation of these two pathways, respectively. CONCLUSIONS: Our results indicate that miRNAs expression profiles in aortic media from TAD were significantly changed. These results may provide important insights into TAD disease mechanisms. This study also suggests that the focal adhesion and MAPK signaling pathways might play important roles in the pathogenesis of TAD.

Downregulation of FHL1 expression in thoracic aortic dissection: implications in aortic wall remodeling and pathogenesis of thoracic aortic dissection.

BACKGROUND: To investigate the potential role of four and a half Lin-11, Isl-1, and Mec-3 (FHL1) protein in the pathogenesis of thoracic aortic dissection (TAD). METHODS: The expression levels and localization of FHL1 protein in aortic tissue of TAD were analyzed using Western blot and immunohistochemistry. Furthermore, small interfering ribonucleic acid was used to knock down the FHL1 gene in rat aortic smooth muscle cells (SMCs). After assessing knockdown efficiency and specificity by real-time polymerase chain reaction and Western blot, the effect of FHL1 knockdown on cell proliferation and apoptosis was evaluated by 3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide assay and flow cytometry, respectively. RESULTS: Compared with normal aortic tissue, FHL1 protein expression in aortic tissue from TAD patients was significantly downregulated. Immunohistochemistry analysis showed that FHL1 was mainly localized in the cytoplasm of SMCs. In diseased aortic tissue, FHL1 immunoreactivity was lowest in SMCs in the split aortic media and adjacent area, but relatively high in SMCs in the aortic intima and adventitia. FHL1 knockdown significantly inhibited the proliferation of rat aortic SMCs but exerted no obvious effect on cell apoptosis. CONCLUSION: FHL1 protein expression is downregulated in TAD. Downregulation of FHL1 expression might contribute to the pathogenesis of TAD, perhaps by suppressing the proliferation of aortic SMCs and affecting aortic wall remodeling.

IGFBP6 Is Downregulated in Unstable Carotid Atherosclerotic Plaques According to an Integrated Bioinformatics Analysis and Experimental Verification.

AIMS: To investigate the differentially expressed genes (DEGs) and molecular interaction in unstable atherosclerotic carotid plaques. METHODS: Gene expression datasets GSE41571, GSE118481, and E-MTAB-2055 were analyzed. Co-regulated DEGs in at least two datasets were analyzed with the enrichment of Gene Ontology Biological Process (GO-BP), Kyoto Encyclopedia of Genes and Genomes (KEGG), protein-protein interaction (PPI) networks, interrelationships between miRNAs/transcriptional factors, and their target genes and drug-gene interactions. The expression of notable DEGs in human carotid artery plaques and plasma was further identified. RESULTS: The GO-BP enrichment analysis revealed that genes associated with inflammatory response, and extracellular matrix organization were altered. The KEGG enrichment analysis revealed that upregulated DEGs were enriched in the tuberculous, lysosomal, and chemokine signaling pathways, whereas downregulated genes were enriched in the focal adhesion and PI3K/Akt signaling pathway. Collagen type I alpha 2 chain (COL1A2), adenylate cyclase 3 (ADCY3), C-X-C motif chemokine receptor 4 (CXCR4), and TYRO protein tyrosine kinase binding protein (TYROBP) might play crucial roles in the PPI networks. In drug-gene interactions, colonystimulating factor-1 receptor had the most drug interactions. Insulin-like growth factor binding protein 6 (IGFBP6) was markedly downregulated in unstable human carotid plaques and plasma. Under a receiver operating characteristic curve analysis, plasma IGFBP6 had a significant discriminatory power (AUC, 0.894; 95% CI, 0.810-0.977), with a cutoff value of 142.08 ng/mL. CONCLUSIONS: The genes COL1A2, ADCY3, CXCR4, and TYROBP are promising targets for the prevention of unstable carotid plaque formation. IGFBP6 may be an important biomarker for predicting vulnerable plaques.

Ankylosing Spondylitis Manifested by Extensive Cervical Erosions with Spontaneous Anterior Atlantoaxial Subluxation.

BACKGROUND: Atlantoaxial instability owing to bone erosions in a patient with ankylosing spondylitis (AS) is rare. We describe the radiographic characteristics, pathology, and treatment of a patient with this rare clinical manifestation and review the literature. CASE DESCRIPTION: A 36-year-old man with an 8-year history of AS presented with progressive neck pain, low back pain, hand numbness, and limited mobility of the neck. Cervical radiography showed anterior atlantoaxial subluxation with bone erosions at the odontoid process and a mass lateral to the atlas and edge of vertebrae. AS was diagnosed according to the modified New York criteria, and the patient underwent a posterior C0-C6 occipitocervical arthrodesis surgery and C3-C6 laminectomy to reconstruct atlantoaxial stability and relieve cervical compression. The symptoms of neck pain and hand numbness improved at the 1-year follow-up, and the patient completely resumed normal activities. Imaging showed realignment of C1-2 with complete decompression of the spinal cord and fusion of the atlantooccipital joint. The internal fixation has remained stable, and progressive bone erosion changes were not found after surgery. CONCLUSIONS: Extensive cervical erosions with spontaneous atlantoaxial subluxation in AS is extremely rare. The erosive change of atlantoaxial bone may be an early feature of AS. Cervical spine radiographs are essential for patients with AS who present with neck pain. Complete decompression and internal fixation are necessary to prevent serious neurologic morbidity from spinal cord injury in such patients.

Hybrid Surgery for Nontaper or Nonstump Lesions in Symptomatic Subacute or Chronic Internal Carotid Occlusion: A Better Solution.

BACKGROUND: Internal carotid artery occlusion (ICAO) causes transient ischemic attack and cerebral infarction. ICAO management remains clinically challenging. We discuss a hybrid treatment combining carotid endarterectomy and endovascular intervention (E-I) for patients with nontaper or nonstump lesions of symptomatic ICAO. METHODS: We treated 32 patients with consecutive nontaper or nonstump ICAO with neurological symptoms with hybrid treatment or E-I. We analyzed the epidemiology, symptoms, angiographic morphology, technical success rate, and perioperative complications. RESULTS: Of the 32 patients, 17 were treated with hybrid surgery and 15, E-I. The demographic data and lesion characteristics were similar between the 2 groups. The overall recanalization success rate was 71.9%. The rate for hybrid surgery was better than that for E-I (88.2% vs. 53.3%). The postoperative cerebral hyperperfusion rate showed no difference between the 2 groups (11.8% vs. 6.7%). Ipsilateral cerebral perfusion improved after treatment. The mean transition time and time to peak were greater than normal (normal values, <6 seconds and <8 seconds, respectively). Both increased significantly after treatment (mean transition time, 11.30 seconds vs. 7.25 seconds; time to peak, 19.30 seconds vs. 15.50 seconds). The incidence of perioperative complications from hybrid surgery was less than that with E-I (5.9% vs. 40.0%). Recurrent cerebrovascular events (5.9% vs. 13.3%) and the 3-month modified Rankin scale score (2.76 ± 0.66 vs. 2.93 ± 0.70) did not differ between the 2 groups. CONCLUSIONS: Recanalization of nontaper or nonstump ICAO with hybrid treatment was more successful than that with E-I, with fewer perioperative complications. The carotid endarterectomy procedure enables easier wire crossing across the occlusion and reduces potential technology-related complications by requiring a shorter lesion and fewer dissections and minimizing the effect of calcification.