NINJ1 Facilitates Abdominal Aortic Aneurysm Formation via Blocking TLR4-ANXA2 Interaction and Enhancing Macrophage Infiltration.

Abdominal aortic aneurysm (AAA) is a common and potentially life-threatening condition. Chronic aortic inflammation is closely associated with the pathogenesis of AAA. Nerve injury-induced protein 1 (NINJ1) is increasingly acknowledged as a significant regulator of the inflammatory process. However, the precise involvement of NINJ1 in AAA formation remains largely unexplored. The present study finds that the expression level of NINJ1 is elevated, along with the specific expression level in macrophages within human and angiotensin II (Ang II)-induced murine AAA lesions. Furthermore, Ninj1flox/flox and Ninj1flox/floxLyz2-Cre mice on an ApoE-/- background are generated, and macrophage NINJ1 deficiency inhibits AAA formation and reduces macrophage infiltration in mice infused with Ang II. Consistently, in vitro suppressing the expression level of NINJ1 in macrophages significantly restricts macrophage adhesion and migration, while attenuating macrophage pro-inflammatory responses. Bulk RNA-sequencing and pathway analysis uncover that NINJ1 can modulate macrophage infiltration through the TLR4/NF-κB/CCR2 signaling pathway. Protein-protein interaction analysis indicates that NINJ1 can activate TLR4 by competitively binding with ANXA2, an inhibitory interacting protein of TLR4. These findings reveal that NINJ1 can modulate AAA formation by promoting macrophage infiltration and pro-inflammatory responses, highlighting the potential of NINJ1 as a therapeutic target for AAA.

Integrative multi-omics analyses reveal vesicle transport as a potential target for thoracic aortic aneurysm.

BACKGROUND: Thoracic aortic aneurysm (TAA) refers to dilation and enlargement of the thoracic aorta caused by various reasons. Most patients have no apparent symptoms in the early stage and are subject to a poor prognosis once the aneurysm ruptures. It is crucial to identify individuals who are predisposed to TAA and to discover effective therapeutic targets for early intervention. METHODS: We conducted a label-free quantitative proteomic analysis among aorta tissue samples from TAA patients to screen differentially expressed proteins (DEPs) and key co-expression modules. Two datasets from Gene Expression Omnibus (GEO) database were included for integrative analysis, and the identified genes were subjected to immunohistochemistry (IHC) validation. Detailed vesicle transport related enrichment analysis was conducted and two FDA-approved drugs, chlorpromazine (CPZ) and chloroquine (CQ), were selected for in vivo inhibition of vesicle transport in mice TAA model. The diameter of thoracic aorta, mortality and histological differences after interventions were evaluated. RESULTS: We found significant enrichments in functions involved with vesicle transport, extracellular matrix organizing, and infection diseases in TAA. Endocytosis was the most essential vesicle transport process in TAA formation. Interventions with CPZ and CQ significantly reduced the aneurysm diameter and elastin degradation in vivo and enhanced the survival rates of TAA mice. CONCLUSIONS: We systematically screened the aberrantly regulated bioprocesses in TAA based on integrative multi-omics analyses, identified and demonstrated the importance of vesicle transport in the TAA formation. Our study provided pilot evidence that vesicular transport was a potential and promising target for the treatment of TAA.

A convenient and effective method for determining organophosphorus pesticides in citrus fruits based on a novel dispersive solid phase extraction using UiO-66/Alg bead as the sorbent.

This work presents a novel, convenient and effective method for assaying organophosphorus pesticides (OPPs) in the pulp and peel of citrus fruits. In this method, shaped UiO-66/alginate (UiO-66/Alg) beads were employed to replace the powder sorbents used in traditional dispersive solid phase extraction (d-SPE) methods. The UiO-66/Alg beads can be easily separated by only using a tweezer within 1 min, which effectively simplifies the sample pretreatment and overcomes the shortages brought by the incomplete separation of powder sorbents. Moreover, the matrix compounds can be effectively excluded by UiO-66/Alg beads, and the UiO-66/Alg beads can be reused at least 8 times. The d-SPE conditions were optimized by a single factor test. The method shows satisfactory sensitivity, accuracy and precision. Furthermore, ATR-FTIR and UV-Vis-DRS were employed to investigate the adsorption mechanism. Finally, the developed method was applied to monitor the OPPs in ten different citrus fruits.

Establishing and Validating a Morphological Prediction Model Based on CTA to Evaluate the Incidence of Type-B Dissection.

Background: Many patients with Type B aortic dissection (TBAD) may not show noticeable symptoms until they become intervention and help prevent critically ill, which can result in fatal outcomes. Thus, it is crucial to screen people at high risk of TBAD and initiate the necessary preventive and therapeutic measures before irreversible harm occurs. By developing a prediction model for aortic arch morphology, it is possible to accurately identify those at high risk and take prompt action to prevent the adverse consequences of TBAD. This approach can facilitate timely the development of serious illnesses. Method: The predictive model was established in a primary population consisting of 173 patients diagnosed with acute Stanford TBAD, with data collected from January 2017 and December 2018, as well as 534 patients with healthy aortas, with data collected from April 2018 and December 2018. Explicitly, the data were randomly separated into the derivation set and validation set in a 7:3 ratio. Geometric and anatomical features were extracted from a three-dimensional multiplanar reconstruction of the aortic arch. The LASSO regression model was utilized to minimize the data dimension and choose relevant features. Multivariable logistic regression analysis and backward stepwise selection were employed for predictive model generation, combining demographic and clinical features as well as geometric and anatomical features. The predictive model's performance was evaluated by examining its calibration, discrimination, and clinical benefit. Finally, we also conducted internal verification. Results: After applying LASSO logistic regression and backward stepwise selection, 12 features were entered into the prediction model. Age, aortic arch angle, total thoracic aorta distance, ascending aorta tortuosity, aortic arch tortuosity, distal descending aorta tortuosity, and type III arch were protective factors, while male sex, hypertension, aortic arch height, and aortic arch distance were risk factors. The model exhibited satisfactory discrimination (AUC, 0.917 [95% CI, 0.890-0.945]) and good calibration in the derivation set. Applying the predictive model to the validation set also provided satisfactory discrimination (AUC, 0.909 [95% CI, 0.864-0.953]) and good calibration. The TBAD nomogram for clinical use was established. Conclusions: This study demonstrates that a multivariable logistic regression model can be used to predict TBAD patients.

Assessing outcomes after thrombectomy with or without iliac vein stenting for young provoked DVT patients with iliac vein stenosis.

BACKGROUND: This study aimed to assess the outcomes of thrombectomy with/without iliac vein stenting for young and transiently provoked DVT patients with iliac vein stenosis. METHODS: This is a retrospective analysis of a prospectively collected multicenter database. Acute, transiently provoked DVT patients between 18 and 45 years old with iliac vein stenosis were included. All patients underwent thrombectomy. Outcomes including the Villalta score, the VEINES-QOL score, and adverse events were evaluated. RESULTS: The data of 522 patients were collected of whom 75 were included, 58 underwent thrombectomy alone (nonstenting group) and 17 underwent thrombectomy and stenting (stenting group). Within 6 months, the Villalta score of patients in stenting group is lower than that of patients in nonstenting group (6 mo: 0.73 ± 0.77 vs. 1.41 ± 0.56, p = .0004), and the VEINES-QOL score of stenting group is higher than that of nonstenting group (6 mo: 89.00 ± 2.94 vs. 87.47 ± 3.72, p = .2141). At the following follow-ups, the Villalta score (12 mo: 0.56 ± 0.49 vs. 0.60 ± 0.58, p = .8266) and VEINES-QOL score (12 mo: 88.36 ± 2.29 vs. 88.31 ± 3.36, p = .9604) between the two groups are similar. CONCLUSION: The stenting group had better efficacy within 6 months after intervention, while there was no significant difference in the symptom, signs, and quality of life between two groups after 6 months within a 2-year follow-up. TRIAL REGISTRATION: This study was registered in the Chinese Clinical Trial Registry (Registration Number: ChiCTR2200056073).

An H2 S-BMP6 Dual-Loading System with Regulating Yap/Taz and Jun Pathway for Synergistic Critical Limb Ischemia Salvaging Therapy.

Critical limb ischemia, the final course of peripheral artery disease, is characterized by an insufficient supply of blood flow and excessive oxidative stress. H2 S molecular therapy possesses huge potential for accelerating revascularization and scavenging intracellular reactive oxygen species (ROS). Moreover, it is found that BMP6 is the most significantly up-expressed secreted protein-related gene in HUVECs treated with GYY4137, a H2 S donor, based on the transcriptome analysis. Herein, a UIO-66-NH2 @GYY4137@BMP6 co-delivery nanoplatform to strengthen the therapeutic effects of limb ischemia is developed. The established UIO-66-NH2 @GYY4137@BMP6 nanoplatform exerts its proangiogenic and anti-oxidation functions by regulating key pathways. The underlying molecular mechanisms of UIO-66-NH2 @GYY4137@BMP6 dual-loading system lie in the upregulation of phosphorylated YAP/TAZ and Jun to promote HUVECs proliferation and downregulation of phosphorylated p53/p21 to scavenge excessive ROS. Meanwhile, laser-doppler perfusion imaging (LDPI), injury severity evaluation, and histological analysis confirm the excellent therapeutic effects of UIO-66-NH2 @GYY4137@BMP6 in vivo. This work may shed light on the treatment of critical limb ischemia by regulating YAP, Jun, and p53 signaling pathways based on gas-protein synergistic therapy.

SENP3 deletion promotes M2 macrophage polarization and accelerates wound healing through smad6/IκB/p65 signaling pathway.

Macrophages preferentially polarize to the anti-inflammatory M2 subtype in response to alterations in the wound microenvironment. SUMO-specific protease 3 (SENP3), a SUMO-specific protease, has been proven to regulate inflammation in macrophages by deSUMOylating substrate proteins, but its contribution to wound healing is poorly defined. Here, we report that SENP3 deletion promotes M2 macrophage polarization and accelerates wound healing in macrophage-specific SENP3 knockout mice. Notably, it affects wound healing through the suppression of inflammation and promotion of angiogenesis and collagen remodeling. Mechanistically, we identified that SENP3 knockout facilitates M2 polarization through the Smad6/IκB/p65 signaling pathway. SENP3 knockout elevated the expression of Smad6 and IκB. Moreover, Smad6 silencing enhanced the expression of p-p65 and proinflammatory cytokines while inhibiting the level of IκB. Our study revealed the essential role of SENP3 in M2 polarization and wound healing, which offers a theoretical basis for further research and a therapeutic strategy for wound healing.

Identification of core cuprotosis-correlated biomarkers in abdominal aortic aneurysm immune microenvironment based on bioinformatics.

Background: The occurrence of abdominal aortic aneurysms (AAAs) is related to the disorder of immune microenvironment. Cuprotosis was reported to influence the immune microenvironment. The objective of this study is to identify cuprotosis-related genes involved in the pathogenesis and progression of AAA. Methods: Differentially expressed lncRNAs (DElncRNAs) and mRNAs (DEmRNAs) in mouse were identified following AAA through high-throughput RNA sequencing. The enrichment analyses of pathway were selected through Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG). The validation of cuprotosis-related genes was conducted through immunofluorescence and western blot analyses. Results: Totally, 27616 lncRNAs and 2189 mRNAs were observed to be differentially expressed (|Fold Change| ≥ 2 and q< 0.05) after AAA, including 10424 up-regulated and 17192 down-regulated lncRNAs, 1904 up-regulated and 285 down-regulated mRNAs. Gene ontology and KEGG pathway analysis showed that the DElncRNAs and DEmRNAs were implicated in many different biological processes and pathways. Furthermore, Cuprotosis-related genes (NLRP3, FDX1) were upregulated in the AAA samples compared with the normal one. Conclusion: Cuprotosis-related genes (NLRP3,FDX1) involved in AAA immune environment might be critical for providing new insight into identification of potential targets for AAA therapy.

m6A Modification Mediates Endothelial Cell Responses to Oxidative Stress in Vascular Aging Induced by Low Fluid Shear Stress.

N6-methyladenosine (m6A) is one of the most prevalent, abundant, and internal transcriptional modification and plays essential roles in diverse cellular and physiological processes. Low fluid shear stress (FSS) is a key pathological factor for many cardiovascular diseases, which directly forces on the endothelial cells of vessel walls. So far, the alterations and functions of m6A modifications in vascular endothelial cells at the low FSS are still unknown. Herein, we performed the transcriptome-wide m6A modification profiling of HUVECs at different FSS. We found that the m6A modifications were altered earlier and more sensitive than mRNA expressions in response to FSS. The low FSS increased the m6A modifications at CDS region but decreased the m6A modifications at 3' UTR region and regulated both the mRNA expressions and m6A modifications of the m6A regulators, such as the RBM15 and EIF3A. Functional annotations enriched by the hypermethylated and hypomethylated genes at low FSS revealed that the m6A modifications were clustered in the aging-related signaling pathways of mTOR, PI3K-AKT, insulin, and ERRB and in the oxidative stress-related transcriptional factors, such as HIF1A, NFAT5, and NFE2L2. Our study provided a pilot view of m6A modifications in vascular endothelial cells at low FSS and revealed that the m6A modifications driven by low FSS mediated the cellular responses to oxidative stress and cell aging, which suggested that the m6A modifications could be the potential targets for inhibiting vascular aging at pathological low FSS.

Adhesive hydrogel wrap loaded with Netrin-1-modified adipose-derived stem cells: An effective approach against periarterial inflammation after endovascular intervention.

Endovascular interventions, such as balloon dilation and stent implantation, are currently recommended as the primary treatment for patients with peripheral artery disease (PAD), greatly improving patient prognosis. However, the consequent lumen restenosis that occurs after endovascular interventions has become an important clinical problem. Inflammation has been proven to be crucial to postoperative restenosis. In previous studies we have identified that Netrin-1-modified adipose-derived stem cells (N-ADSCs) transplantation is an effective anti-inflammatory strategy to repair vascular damage. Nevertheless, it remained to be explored how one could constantly deliver N-ADSCs onto damaged arteries. Therefore, we developed an adhesive double network (DN) hydrogel wrap loaded with N-ADSCs for sustained perivascular delivery. Inspired by the adhesion mechanism of mussels, we developed an adhesive and tough polyacrylamide/calcium-alginate/reduced graphene oxide/polydopamine (PAM/CA/rGO/PDA) hydrogel. Dopamine was attached to graphene sheets and limitedly oxidized to generate free catechol groups. The hydrogel could wrap damaged arteries and induce anti-inflammatory effects through N-ADSCs. In vitro experiments demonstrated that N-ADSCs significantly promoted the M2 polarization of macrophages to anti-inflammatory phenotypes and reduced the expression of inflammatory factors. In vivo experiments in a rat carotid artery guidewire injury model showed that the adhesive hydrogel wrap loaded with N-ADSCs could significantly reduce arterial inflammation, inhibit intimal hyperplasia and improve re-endothelialization. Altogether, this newly developed N-ADSCs-loaded hydrogel wrap provides an effective slow-releasing system, which may be a promising way to prevent and treat restenosis after endovascular interventions.

Exosomal miR-17-5p from adipose-derived mesenchymal stem cells inhibits abdominal aortic aneurysm by suppressing TXNIP-NLRP3 inflammasome.

BACKGROUND: Preclinical studies have suggested that adipose-derived mesenchymal stem cells (ADSCs) transplantation can suppress abdominal aortic inflammation and aneurysm expansion through paracrine factors. Yet, the mechanism of action is not fully understood. In the present study, we further examined the function and mechanism of ADSC-derived exosomes (ADSC-exos) and their microRNA-17-5p (miR-17-5p) on the abdominal aortic aneurysm (AAA) progression. METHODS: ADSC-exos were isolated and identified. DiR and PKH67 staining were used to trace ADSC-exo in vivo and in vitro. Raw264.7 cells were applied to perform in vitro experiments, while a murine AAA model induced using angiotensin II (Ang II) was used for in vivo testing. The expression level of miR-17-5p in macrophages and Ang II-treated macrophages after ADSC-exos treatment was determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The target relation between miR-17-5p and thioredoxin-interacting protein (TXNIP) was identified by a dual-luciferase reporter gene assay. Artificial activation and block of experiments of miR-17-5p and TXNIP were conducted to clarify their functions in inflammation during AAA progression. The severity of AAA between groups was assessed by maximal aorta diameter, AAA incidence, survival rate, and histological stainings. Besides, inflammasome-related proteins and macrophage pyroptosis were further evaluated using western blot, RT-qPCR, and enzyme-linked immunosorbent assay (ELISA). RESULTS: The ADSC-exos were isolated and identified. In vivo testing showed that ADSC-exos were mainly distributed in the liver. Meanwhile, in vitro experiments suggested that ADSC-derived exosomes were taken up by macrophages, while inside, ADSC-exos miR-17-5p decreased a TXNIP induced by Ang II by directly binding to its 3'-untranslated region (3'UTR). Furthermore, overexpression of miR-17-5p enhanced the therapeutic function of ADSC-exos on inflammation during AAA expansion in vivo, while its inhibition reversed this process. Finally, overexpressed TXNIP triggered macrophage pyroptosis and was alleviated by ADSC-derived exosomes in vitro. CONCLUSION: ADSC-exos miR-17-5p regulated AAA progression and inflammation via the TXNIP-NLRP3 signaling pathway, thus providing a novel insight in AAA treatment.

Clinical Validation of the Impact of Branch Stent Extension on Hemodynamics in ISF-TEVAR Involving LSA Reconstruction.

Background: The study of hemodynamics regarding thoracic endovascular aortic repair (TEVAR) is helpful to improve the surgical efficacy. Objective: Correlations between hemodynamic changes and branch stent extension length and interference factors for branch stent extension length of in situ fenestration TEVAR (ISF-TEVAR) involving the left subclavian artery (LSA) were evaluated. Materials and Methods: This study retrospectively analyzed 196 patients with Stanford type B aortic dissection who received in situ laser fenestrated thoracic endovascular aortic repair with LSA fenestration from April 2014 to March 2021. Branch stent extension to the main stent graft was evaluated by the computed tomographic angiography (CTA). Hemodynamic change of LSA was defined as a 20 mmHg interbrachial systolic pressure difference. The factors affecting the extension of the branch stent were also evaluated. Results: All patients underwent ISF-TEVAR with LSA fenestration, and there was no recurrence during the follow-up. The mean length of the branch stent extension was 10.37 ± 0.34 mm, which was used to divide the patients into long and short groups. Asymptomatic hemodynamic changes (defined as a 20 mmHg interbrachial systolic pressure difference) in LSA were observed in 61 patients undergoing ISF-TEVAR involving LSA fenestration. The Spearman correlation analysis showed extension length of a branch stent >1.5 cm elevated the risk of hemodynamic changes. Conclusion: Overall, we conclude that branch stent extension length >1.5 cm induced LSA hemodynamic changes. Appropriate shortening of the stent extension length can improve the curative effect of ISF-TEVAR, especially when faced with a type II/III aortic arch and stent angles of <30 degrees.

Endovascular Excimer Laser-Assisted Balloon Angioplasty for Infrapopliteal Arteries in Thromboangiitis Obliterans: A Treatment for Acute-Phase TAO.

Background: Thromboangiitis obliterans (TAO, Buerger's disease) is an inflammatory and obstructive vasculopathy, which leads to limb ischemic rest pain and ulcerations in the acute stage. Objectives: This study aimed to assess the feasibility of excimer laser-assisted balloon angioplasty (BA) for patients with acute infrapopliteal TAO. Method: This was a single-center retrospective cohort study. In this study, 220 patients with 210 target limbs between January 2012 and September 2021 were involved. Among them, 52 target limbs have received endovascular excimer laser-assisted balloon angioplasty from January 2017. The ankle brachial index (ABI), rest pain score, ulcer, Rutherford classification, and TASC II classification were assessed. The follow-up time was 6 months. Results: The technical success rate of laser + BA and BA groups was 71.15 and 65.82% (p = 0.5021), respectively. After intervention, the ABI of two groups were 0.85 ± 0.20 and 0.77 ± 0.20 (p = 0.0419), and the rest pain score was 1.00 ± 1.43 and 1.71 ± 2.25 (p = 0.0449). During the 6 months follow-up, the ABI of two groups was 0.76 ± 0.17 and 0.75 ± 0.23 (p = 0.8539), the rest pain score was 1.43 ± 1.82 and 2.24 ± 2.06 (p = 0.0783), and the ulcer rate was 23.68 and 40.98% (p = 0.0867), respectively. The proportion of patients who were assessed as TASC II C/D or Rutherford 4-6 in laser +BA group was significantly lower than that in BA group, indicating that the former had better efficacy. The rate of critical limb ischemia and restenosis in the laser +BA group was lower than that in the BA group (47.36 vs. 67.22%; 21.05 vs. 34.43%) during follow-up. In the laser + BA group, the reintervention rate was lower than that in the BA group (2.70 vs. 8.20%, p = 0.0425). No serious adverse events (AEs) occurred. Conclusion: Excimer laser debulking-assisted angioplasty is a feasible, effective, and safe method to treat acute infrapopliteal TAO.

Comprehensive insights on pivotal prognostic signature involved in clear cell renal cell carcinoma microenvironment using the ESTIMATE algorithm.

Emerging evidence has highlighted that the immune and stromal cells formed the majority of tumor microenvironment (TME) which are served as important roles in tumor progression. In our study, we aimed to screen vital prognostic signature associated with TME in clear cell renal cell carcinoma (ccRCC). We obtained total 611 samples from TCGA database consisting of transcriptome profiles and clinical data. ESTIMATE algorithm was applied to estimate the infiltrating fractions of immune/stromal cells. We found that the immune scores revealed more prognostic significance in overall survival and positive associations with risk clinical factors than stromal scores. We carried out differential expression analysis between Immunescore and stromalscore groups to obtain the 72 intersect genes. Protein to protein interaction (PPI) network and functional analysis was performed to indicate potential altered pathways. Additionally, we further conducted multivariate Cox analysis to identify 12 hub genes associated highly with TME of ccRCC using a stepwise regression procedure. Accordingly, risk score was constructed from the multivariate Cox results and Receiver Operating Characteristic (ROC) curve was used to assess the predictive value (AUC = 0.781). The ccRCC patients with high risk scores suffered poor survival outcomes than that with low risk scores. In the validation cohort from GSE53757, TNFSF13B, CASP5, and GJB6 correlated positively with tumor stages, while FREM1 negatively correlated with tumor stages. Importantly, we further observed that TNFSF13B, CASP5 and XCR1 showed the remarkable correlations with tumor-infiltrating immune cells. Taken together, our research identified specific signatures that related to the infiltration of stromal and immune cells in TME of ccRCC using the transciptome profiles, which reached a comprehensive understanding of tumor microenvironment in ccRCC.

Identification of five long noncoding RNAs signature and risk score for prognosis of bladder urothelial carcinoma.

Nowadays, an increasing number of studies illustrated that bladder urothelial cancer (BLCA) may act as the most common subtype of urological malignancies with a high rate of recurrence and metastasis. In this study, we attempted to establish a prognostic model and identify the possible pathway crosstalk. Long noncoding RNAs (lncRNAs) and mRNA expression and corresponding clinical information of patients with BLCA were downloaded from The Cancer Genome Atlas (TCGA). The differentially expressed genes analysis, univariate Cox analysis, the least absolute shrinkage, and selection operator Cox (LASSO Cox) regression model were then applied to identify five crucial lncRNAs (AC092725.1, AC104071.1, AL023584.1, AL132642.1, and AL137804.1). The multivariate cox analysis was utilized to calculate the regression coefficients (ßi ). The risk-score model was subsequently constructed as follows: (0.13541AC092725.1) + (0.20968AC104071.1) + (0.1525AL023584.1) - (0.14768AL132642.1) + (0.14387AL137804.1). Nomogram and assessment of overall survival (OS) prediction were verificated by the receiver operating characteristic curve in the testing group. As to 3-, 5-year OS prediction, the area under curve (AUC) for the nomogram of training data set was 0.83 and 0.86. Besides, the AUC (0.883 and 0.879) presented excellent predictive power in the testing group. In addition, the calibration plots validated the predictive performance of the nomogram. Weighted correlation network analysis (WGCNA) coupled with functional enrichment analysis contributed to explore the potential pathways, including PI3K-Akt, HIF-1, and Jak-STAT signaling pathways. Construction of the risk-score model and data analysis were both derived from multiple packages on the basis of the R platform chiefly.