O-GalNAc glycosylation, a prevalent and important post-translational modification of proteins, participates in a variety of physiological and pathological processes including development, immune recognition, and tumor progression. Therefore, systematic analysis of O-GalNAc glycoproteins is crucial for elucidating its regulatory mechanisms and discovering novel disease biomarkers. However, O‑GalNAc glycoproteomic research remains highly challenging due to the inherently low abundance of glycoproteins, the high heterogeneity of glycans, and the frequently dense clustering of glycosites. In recent years, significant progress has been made in the development of glycopeptide enrichment techniques such as lectin affinity chromatography, chemoenzymatic method, and metabolic labeling, along with advances in mass spectrometry fragmentation strategies and dedicated bioinformatics tools for O-glycoproteomic data analysis. These developments have collectively enabled researchers to study protein O-GalNAc glycosylation in a more systematic and in-depth manner. In this review, we summarize recent advances in mass spectrometry-based O-GalNAc glycoproteomic technologies, evaluate the latest analytical software tools, and discuss future perspectives in the field.

We performed a mixed-cohort study to establish the scalp lipid–microbe functional linkages in dandruff using untargeted quantitative lipidomic and shot-gun metagenomic technology. Our findings suggest that suppressing scalp microbes facilitates lipid removal and alleviates dandruff more effectively.

Abdominal aortic aneurysm (AAA) is a fatal cardiovascular disease with hidden onset and high risk of rupture death, which is a major public health concern worldwide. Notably, aortic adventitia is the earliest lesions and the last barrier to rupture, and adventitial fibroblasts (AFs) are the key cellular components of aortic adventitia. Current treatments mainly focus on surgical resection with prosthetic graft replacement and endovascular stent grafting, yet failed to reconstruct the adventitia and abdominal aorta's natural structure effectively. In this study, we proposed an extravascular synergistic cocktail (ESC) therapeutic strategy to treat AAA by incorporating mechanical supporting and vascular remodeling inhibition based on a self-healing bioelastomer with antifibrotic drug of lanifibranor. The single-cell RNA-sequencing revealed potential myogenic transformation of AFs in AAA with the release of chemokines like monocyte chemoattractant protein-1 (MCP-1). The underlying mechanism of lanifibranor in maintaining the phenotype of AFs via promoting lipid metabolism was disclosed. The bioelastomer with multiple reversible dynamic bonds exhibits remarkable arterial-like mechanical properties and ultra-fast self-healing ability under blood condition, and enables rapid in situ AAA wrapping treatment. In both the rat and dog AAA models, it has been fully demonstrated that the ESC therapy can provide effective mechanical support to prevent vascular dilation while continuously releasing lanifibranor to regulate myogenic transformation of AFs to inhibit the development of AAA. Additionally, we investigated the feasibility of minimally invasive laparoscopic implantation in bama pigs, emphasizing its potential for clinical translation and application.

Interventional therapy and surgery play important roles in the treatment of various diseases, but they cause varying degrees of vascular injury. Currently, the side effects are often overlooked. Here, we observed abnormal nuclear morphology (nuclear dysmorphism) and vascular aging in injured human and rodent arteries. Platelet-derived microvesicles (PMVs) adhere to injured blood vessels, leading to nuclear dysmorphism and cell senescence in vascular smooth muscle cells (VSMCs). This occurs because PMV adherence reduces intracellular Zn2+ levels, which impairs Zn2+-dependent processing of prelamin A by the enzyme ZMPSTE24. Consequently, prelamin A accumulates in VSMCs, contributing to the observed nuclear dysmorphism and cell senescence. RNA sequencing and loss-of-function assays revealed that Zinc transporter solute carrier family 39 member 4 (SLC39A4, also called ZIP4) deficiency accounts for the decreased Zinc concentration. Consistently, Zmpste24(+/-) and Zmpste24(-/-) mice displayed significant cumulative prelamin A, deteriorated nuclear dysmorphism and vascular aging. Whole genome bisulfite sequencing (WGBS) and bioinformatic analysis illustrated that demethylation of genes within Lamina-associated domains (LADs) participates in nuclear dysmorphism and cell senescence. Of note, Zinc supplementation, especially using platelet membrane-coated Zn-MOF nanoparticles, robustly alleviated nuclear dysmorphism and vascular aging. Our data established a novel and significant role of pMVs/ZIP4/zinc/prelamin A axis in promoting nuclear dysmorphism and vascular aging after injury.

Peripheral nerve injury (PNI) affects over 20 million individuals globally each year, with incomplete nerve regeneration presenting a significant socioeconomic burden, particularly in diabetic patients. M2 macrophages play a crucial role in nerve regeneration by promoting tissue remodeling and repair. However, the post-injury microenvironment impedes the stabilization of the M2 phenotype, limiting their therapeutic potential. In vitro experiments demonstrate that macrophages modified with Netrin-1 can be polarized toward the M2 phenotype. Exosomes secreted by these Netrin-1-modified macrophages (N-Exos) significantly enhance endothelial cell proliferation, migration, tube formation, and survival under hyperglycemic conditions. The underlying mechanism is likely mediated by the activation of the PI3K/AKT/mTOR pathway through Unc5b. To further enhance the therapeutic effects of N-Exos, we incorporated these exosomes into a berberine-modified chitosan-based hydrogel (NE@CSB), thereby developing an innovative synergistic therapeutic approach. The NE@CSB hydrogel demonstrates favorable injectability, thermoresponsive sol-gel transition, strong tissue adhesion, and sustained release of exosomes. In the sciatic nerve injury model of diabetic mice, blood flow perfusion and histological analysis of injured nerves demonstrated that NE@CSB hydrogel could effectively enhanced angiogenesis and sciatic nerve regeneration. Overall, NE@CSB presents a promising and safe therapeutic strategy for the clinical treatment of diabetic PNI.

Background: The optimal design concept and the safety of drug-coated balloons (DCBs) have not been completely determined yet. Also, the optimal approach for DCB-based revascularization of peripheral artery diseases still remains undefined. This study was designed to explore the in vivo pharmacokinetic and histopathological effects of DCBs using Ranger and 4 Chinese DCBs after implantation and administration in New Zealand rabbits. Materials and Methods: Fifty New Zealand rabbits were divided into 5 groups with 10 rabbits in each group according to the DCB used: Ranger (Boston Scientific), Orchid (Acotec), Reewarm (Endovastec), Ultrafree (Zylox), and Yaohang (Polyrey). After being guided to the lower segment of the abdominal aorta, the DCB was inflated for 3 minutes. Plasma, inflated infrarenal aorta, vastus lateralis muscle, anterior tibial muscle, and right toes were harvested for histological and paclitaxel concentration analyses 4 hours or 28 days after the angioplasty. Results: At 4 hours after aortic angioplasty, the overall paclitaxel concentrations in aortic wall were not statistically different (p = 0.050), but the paclitaxel concentrations in vastus lateralis muscle (p = 0.002), anterior tibial muscle (p = 0.006) and toe (p < 0.001) were not totally same according to the results of Kruskal-Wallis test. In toe, concentrations of paclitaxel were significantly lower for the Ranger (120.8 ng/g) DCB than for the Orchid (1880 ng/g; p = 0.008), Reewarm (347 ng/g; p = 0.016), and Ultrafree (261 ng/g; p = 0.016) DCBs. Concentrations of paclitaxel in the toe were not statistically different between Ranger and Yaohang DCBs (p = 0.421). Neointimal area (p < 0.001), neointimal thickness (p < 0.001), and percentage of luminal stenosis (p < 0.001) were less for Ranger DCB than for other DCBs 28 days after aortic angioplasty. The differences in paclitaxel concentrations in tissues 28 days after aortic angioplasty were not statistically significant. Conclusion: Different design concepts will make a difference in the in vivo pharmacokinetic and histopathological effects of DCBs. The Ranger DCB can achieve similar drug delivery efficiency as other higher-dose DCBs and fewer neointimal hyperplasia. Although clinical implications remain to be further investigated, the present results may provide implications for the design and use of DCBs. Clinical Impact The current preclinical study suggested that different design concepts would make a difference in the pharmacokinetic and histopathological effects of drug-coated balloons (DCBs). Though Ranger DCB had the lowest paclitaxel loading in this study, it still achieved similar drug delivery efficiency with other higher-dose DCBs. Also, neointimal hyperplasia was less for the Ranger DCB than for the other DCBs 28 days after aortic angioplasty. Although clinical implications remain to be further investigated, the present results may provide implications for the design and use of DCBs.

Abdominal aortic aneurysm (AAA) is a chronic inflammation-driven disease characterized by aortic wall destruction and expansion, leading to high morbidity and mortality. However, previous drug treatments for its common risk factors have not achieved favorable results, and the early prevention and treatment is still the main clinical dilemma. Anti-inflammation therapy is a promising therapeutical method targeting its pathogenesis mechanism, but it has not been explored in depth. Herein, interleukin-1 receptor antagonist-loaded manganese-doped mesoporous silica nanoparticles (IL-1Ra@MMSN) were designed and synthesized to target macrophage-mediated chronic aortic inflammation for AAA treatment. IL-1Ra@MMSN showed high IL-1Ra-loading efficiency, great stability and pH-responsive drug-releasing property. IL-1Ra@MMSN specially phagocytosed by macrophages can protect against oxidative stress injury and promoted the M2 polarization via transforming growth factor-β (TGF-β) signaling in vitro. Furthermore, IL-1Ra@MMSN exhibited good lesion targeting ability, hemocompatibility and biocompatibility in angiotensin II-induced murine AAA model. In vivo experiments also confirmed the excellent treatment efficacy in reducing AAA formation and progression via protecting aortic wall integrity and promoting anti-inflammatory microenvironment. Taken together, the current study demonstrated that IL-1Ra@MMSN is a promising nanoplatform for early intervention of AAA, which provides a novel treatment strategy based on anti-inflammatory immune regulation.

There remains clinical uncertainty concerning the relationship between systemic inflammation and subsequent abdominal aortic aneurysm (AAA) risk. To investigate the association between chronic systematic inflammation markers (C-reactive protein (CRP), peripheral immune cell counts, and their derived ratios) and risk of AAA incidence, and identify potential effect modifiers. We included 271,068 individuals from the UK Biobank, who were free of aortic aneurysm and other conditions impacting their inflammatory states at baseline. Cox proportional-hazards model was used to analyze associations between inflammatory biomarkers and AAA, adjusting for AAA polygenetic risk score (PRS) and major risk factors. Restricted cubic splines were plotted to visualize non-linear relationship. Subgroup analyses by age, sex, hypertension, smoking and PRS were performed to identify any interaction. Over a median follow-up of 13·9 years, 629 incident AAAs were recorded. For each 1-SD increase in baseline CRP, lymphocyte, monocyte, and neutrophil counts, the risk of AAA increased by 46

Hydrogel nanofibers provide a regeneration-permissive environment conducive to the regrowth of numerous nerve fibers, thereby enhancing regenerative capacity in cases of peripheral nerve injury and spinal cord injury. However, developing hydrogel nanofiber-based nerve guidance conduits (NGCs) with tailored drug release profiles to synergistically promote angiogenesis and axonal regeneration remains a significant challenge. In this study, novel polydopamine (PDA)-modified gelatin methacryloyl (GelMA) hydrogel nanofibers are developed as an efficient drug delivery platform for sustained release of Secreted Frizzled-Related Protein-2 (SFRP2). This platform aims to promote neurite outgrowth, facilitate nerve function recovery, and enhance angiogenesis through Wnt signaling pathways. Results indicate that PDA coating significantly improves the hydrophilicity and mechanical properties of GelMA hydrogel nanofibers, which were fabricated using a combination of electrospinning and photo-crosslinking technology. This modification enables SFRP2 loading for sustained release through π-π stacking interactions and hydrogen bonding. In vitro experiments demonstrate that SFRP2-loaded hydrogel nanofibers effectively enhance the adhesion, proliferation, viability, and migration of Mouse Schwann Cells (MSCs), while also promoting tube formation and ameliorating the inflammatory microenvironment of Human Umbilical Vein Endothelial Cells (HUVECs). Furthermore, the SFRP2-loaded hydrogel nanofibers are confirmed to exert their functions for angiogenesis and peripheral nerve regeneration via the calcium-dependent calcineurin/NFATc3 signaling pathway. Finally, the hydrogel nanofiber-based NGCs are applied in a mouse model of peripheral nerve injury, and results demonstrate that the SFRP2 ~ PDA@GelMA conduit significantly enhances angiogenesis, promotes peripheral nerve repair, and facilitates target muscle restoration and functional recovery, thus presents a promising therapeutic strategy for patients with peripheral nerve injuries. We developed novel polydopamine (PDA)-modified gelatin methacryloyl (GelMA) hydrogel nanofibers for sustained Secreted Frizzled-Related Protein 2 (SFRP2) release to promote peripheral nerve regeneration and angiogenesis synchronously. In vitro experiments demonstrated that these nanofibers significantly enhanced Schwann cell migration and endothelial cell tube formation. In a mouse model of peripheral nerve injury, the SFRP2-loaded nanofibers effectively promoted angiogenesis, nerve repair, and target muscle restoration via the calcineurin/NFATc3 signaling pathway. These findings suggest a promising therapeutic strategy for peripheral nerve injuries.

Atherosclerosis is the main pathogenic factor of various cardiovascular diseases. During the pathogenesis of atherosclerosis, macrophages play a major role, mainly by secreting pro-inflammatory cytokines and taking in lipids to form foam cells. Thiamine pyrophosphate (TPP) is an antagonist of the P2Y6 receptor, which is overexpressed on macrophages during atherosclerosis and facilitates the lipid phagocytosis of macrophages. However, the excessive accumulation of TPP may interfere with some vital metabolic processes like the tricarboxylic acid cycle, oxidative phosphorylation and the pentose phosphate pathway. Herein, we designed and constructed a nanoparticle ZIF-8@TPP for the treatment of atherosclerosis. The as-established ZIF-8@TPP nanoplatform exhibited specific cytotoxicity towards macrophages in vitro. Meanwhile, histological analysis confirmed the excellent therapeutic efficacy of ZIF-8@TPP in vivo. Mechanistic studies indicated that ZIF-8@TPP potentially lowered lipid phagocytosis and lipid metabolism of macrophages via the PI3K/AKT/MSR1 pathway. This study also demonstrated that the anti-atherosclerotic effect of TPP was enhanced after combination with a prototypical metal-organic framework (MOF), ZIF-8. This synergistic controlled-release drug delivery system may provide a novel idea for anti-atherosclerosis therapy by combining reagents that can inhibit lipid phagocytosis of macrophages with MOFs.

Background: Despite appropriate treatment, up to 50% of patients with proximal deep vein thrombosis will develop postthrombotic syndrome (PTS). Once PTS occurs, there is no specific treatment, and some patients constantly experience intolerable symptoms. Hence, prevention of PTS is important. Objectives: To characterize vein wall remodeling after thrombus and investigate the effects of antiproliferative agent on postthrombotic vein wall remodeling in murine and human subjects. Methods: Features of postthrombotic vein wall remodeling in murine and human subjects were characterized using imaging and histologic examinations. Paclitaxelloaded hydrogels were used to assess the effects of antiproliferative agent on the remodeling in murine model. Based on the abovementioned results, a pilot study was conducted to assess the effects of paclitaxel-coated balloon dilation in patients with severe PTS experiencing intolerable symptoms. The control cohort was obtained by 1:1 propensity score matching from a prospective database. Results: Structural and functional alterations in postthrombotic vein wall were verified by imaging and histologic examinations, and predominant active alpha-smooth muscle actin-positive cells and fibroblast-specific protein 1-positive cells proliferation was observed. In the murine model, the application of paclitaxel-loaded hydrogels inhibited the remodeling. In the pilot clinical study, patients receiving drug-coated balloon demonstrated benefits in Villalta scores and venous clinical severity scores compared with those not receiving drug-coated balloon, and no severe adverse events were reported except for thrombosis recurrence. Conclusion: Cell proliferation plays an important role in postthrombotic vein wall remodeling. Inhibition of cell proliferation inhibits the remodeling in murine model and may reduce signs and symptoms in patients with severe PTS.

Objectives Aortic dissection (AD) is a life-threatening condition that requires intensive care and management. This paper explores the role of fluid management in the clinical care of AD patients, which has been unclear despite the substantial existing research that has been conducted on the treatment of AD.Design A retrospective case-control study using data for AD patients from public databases.Setting Two public intensive care unit (ICU) databases with hospital courses from the USA, Medical Information Mart for Intensive Care (MIMIC)-IV critical care dataset and the eICU Collaborative Research Database, with data from 2008 to 2019.Participants A total of 751 adult AD patients with detailed fluid management records from two databases were included.Interventions The mean 24-hour intake and output were calculated by dividing the total amount of intake and output by the number of days in the ICU, respectively. The mean 24-hour fluid balance was generated by subtracting the output from the intake.Outcome measures The relationship between the mean 24-hour fluid management and all-cause in-hospital death was assessed through univariate and multivariable regression analyses.Results A positive correlation was found between mean 24-hour fluid intake and in-hospital mortality among AD patients (OR 1.029, 95% CI (1.018, 1.041), p<0.001), whereas a negative correlation was revealed between mean 24-hour fluid output and in-hospital mortality (OR 0.941, 95% CI (0.914, 0.968), p<0.001). A similar result was found for mean 24-hour fluid balance (OR 1.030, 95% CI (1.019, 1.042), p<0.001), and the cut-off was selected to be 5.12 dL (AUC=0.778, OR 3.066, 95% CI (1.634, 5.753), p<0.001).Conclusions This study stresses the importance of fluid balance in the clinical care of AD patients and provides new insights for optimising fluid management and monitoring strategies beyond the conventional focus on blood pressure and heart rate management.

Objective Aortic dissection (AD) is a severe emergency with high morbidity and mortality, necessitating strict monitoring and management. This retrospective study aimed to identify prognostic factors and establish predictive models for in-hospital mortality among AD patients in the intensive care unit (ICU). Methods We retrieved ICU admission records of AD patients from the Medical Information Mart for Intensive Care (MIMIC)-IV critical care data set and the eICU Collaborative Research Database. Functional data analysis was further applied to estimate continuous vital sign processes, and variables associated with in-hospital mortality were identified through univariate analyses. Subsequently, we employed multivariable logistic regression and machine learning techniques, including simple decision tree, random forest (RF), and eXtreme Gradient Boosting (XGBoost) to develop prognostic models for in-hospital mortality. Results Given 643 ICU admissions from MIMIC-IV and 501 admissions from eICU, 29 and 28 prognostic factors were identified from each database through univariate analyses, respectively. For prognostic model construction, 507 MIMIC-IV admissions were divided into 406 (80%) for training and 101 (20%) for internal validation, and 87 eICU admissions were included as an external validation group. Of the four models tested, the RF consistently exhibited the best performance among different variable subsets, boasting area under the receiver operating characteristic curves of 0.870 and 0.850. The models highlighted the mean 24-h fluid intake as the most potent prognostic factor. Conclusions The current prognostic models effectively forecasted in-hospital mortality among AD patients, and they pinpointed noteworthy prognostic factors, including initial blood pressure upon ICU admission and mean 24-h fluid intake.

Tumor-associated MUC1 is coated with a high density of O-GalNAc glycans, which are initiated by a family of polypeptide N-acetyl-alpha-galactosaminyltransferases (GalNAc-Ts). However, the O-glycosylation process of MUC1 by each GalNAc-T isoform remains unclear. Here, we successfully obtained 14 human GalNAc-Ts with high catalytic activity based on a bacterial expression system. Employing MUC1-derived peptides as substrates, we systematically investigated the catalytic properties and site specificity of these GalNAc-Ts by chromatography and mass spectrometry, and found that they could be classified into two clusters. These two GalNAc-T clusters initially catalyze the threonine residue within GSTA or GVTS motifs, respectively, resulting in high O-glycosylation occupancy of both motifs. Moreover, molecular dynamics simulations and site-directed mutagenesis confirmed that the initial O-glycosite preference of GalNAc-Ts on MUC1 is controlled by two critical residues within the peptide-binding pocket. Swapping of the corresponding residues between two GalNAc-T clusters could exchange their initial O-glycosite preference. Quantum mechanics calculations further revealed the detailed catalytic mechanisms of GalNAc-Ts. Our work contributes to understanding the catalytic synthesis of multisite O-glycosylation of MUC1 by GalNAc-Ts, facilitating the development of O-glycosite-specific MUC1 vaccines.

>Small cell lung cancer(SCLC) is a recalcitrant cancer featured with high metastasis capability. We previously found that TAZ acts as a crucial molecular switch in orchestrating SCLC phenotypic transition and metastasis(Jin et al., 2022). However, the downstream mechanisms by which TAZ regulates SCLC malignant progression remain elusive.Global down-regulation of microRNAs is commonly observed in human cancers(Parayath et al., 2022). Hippo pathway components YAP/TAZ have been reported to regulate miRNA biogenesis through the microprocessor and DICER complexes(Chaulk et al., 2014; Mori et al., 2014). To identify the downstream miRNAs of TAZ involved in regulating SCLC malignant progression, we utilized small RNA sequencing and compared the down-regulated miRNAs in SCLC metastasizing cells(SMC) vs. non-SMC and non-SMC-shT az vs.non-SMC-shL uc. Thirteen miRNAs were commonly downregulated in both the SMC and non-SMC-shT az groups(Fig. 1A;Table S1).

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.

Messenger RNA vaccines lack specificity for dendritic cells (DCs)-the most effective cells at antigen presentation. Here we report the design and performance of a DC-targeting virus-like particle pseudotyped with an engineered Sindbis-virus glycoprotein that recognizes a surface protein on DCs, and packaging mRNA encoding for the Spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or for the glycoproteins B and D of herpes simplex virus 1. Injection of the DC-targeting SARS-CoV-2 mRNA vaccine in the footpad of mice led to substantially higher and durable antigen-specific immunoglobulin-G titres and cellular immune responses than untargeted virus-like particles and lipid-nanoparticle formulations. The vaccines also protected the mice from infection with SARS-CoV-2 or with herpes simplex virus 1. Virus-like particles with preferential uptake by DCs may facilitate the development of potent prophylactic and therapeutic vaccines.