Importance A notable proportion of patients with ankylosing spondylitis (ie, radiographic axial spondyloarthritis [r-axSpA]) experience either inadequate responses or intolerance to available therapies, highlighting the need for additional treatment options that offer sustained disease control. Objective To evaluate the efficacy and safety of vunakizumab, a novel anti–interleukin (IL) 17A monoclonal antibody, in r-axSpA. Design, Setting, and Participants This randomized, double-blind, placebo-controlled, phase 2 to 3 clinical trial was conducted in 38 hospitals in China. A total of 548 adult patients (aged ≥18 years) with active r-axSpA were enrolled between June 9, 2021, and March 30, 2023. Data were analyzed from November 3, 2023, to January 14, 2024. Interventions In phase 2, patients were randomized 2:2:1 to receive a subcutaneous dose of vunakizumab, 120 mg, vunakizumab, 240 mg, or placebo; vunakizumab, 120 mg was determined to be the recommended dose after interim analysis. In phase 3, patients were randomized 2:1 to receive vunakizumab, 120 mg or placebo at weeks 0, 2, 4, 8, and 12; from week 16, all patients received vunakizumab, 120 mg every 4 weeks through week 32. Main Outcomes and Measures The primary end point was the proportion of patients with improvement of 20% or greater in Assessment of Spondyloarthritis International Society response criteria (ASAS20) at week 16. Secondary end points were ASAS20 at week 32, improvement of 40% or greater in ASAS response criteria (ASAS40) at weeks 16 and 32, improvement of 20% or greater in 5 of 6 ASAS domains (ASAS5/6) at weeks 16 and 32, and changes from baseline at weeks 16 and 32 in Bath Ankylosing Spondylitis Disease Activity Index score, Bath Ankylosing Spondylitis Functional Index score, Bath Ankylosing Spondylitis Metrology Index score, 36-Item Short Form Survey score, and Ankylosing Spondylitis Quality of Life Questionnaire score. Results During the entire study, 548 patients were randomized (mean [SD] age, 33.0 [8.9] years; 440 [80.3%] male), including 294 receiving vunakizumab, 120 mg and 146 receiving placebo. At week 16, the ASAS20 response rate was significantly higher with vunakizumab, 120 mg vs placebo (193 [65.6%] vs 62 [42.5%], respectively; difference, 23.2% [95% CI, 11.8%-34.0%]; P < .001); the ASAS40 response rate also favored vunakizumab, 120 mg vs placebo (136 [46.3%] vs 35 [24.0%], respectively; difference, 22.3% [95% CI, 13.3%-31.3%]; P < .001). Responses with vunakizumab were sustained through 32 weeks. During the placebo-controlled period, incidence of adverse events (246 [83.7%] vs 119 [81.5%]) were comparable in the vunakizumab, 120 mg and placebo groups. Conclusions and Relevance In this randomized clinical trial of patients with r-axSpA, vunakizumab, 120 mg significantly improved signs and symptoms of r-axSpA at week 16 compared with placebo, with sustained efficacy through 32 weeks, and demonstrated a tolerable safety profile. These findings support vunakizumab, 120 mg as a new treatment option for active r-axSpA. Trial Registration ClinicalTrials.gov Identifier: NCT04840485

Introduction Rheumatoid Arthritis (RA) is a chronic inflammatory disease often linked to Interstitial Lung Disease (ILD). Slit guidance Ligand-3 (Slit3), a member of the axon guidance molecule superfamily, mediates a series of biological activities, including the regulation of the inflammatory response and fibrosis. This study explored Slit3’s clinical significance as a serum biomarker for RA-ILD. Methods Between November 2022 and April 2024, patients diagnosed with RA were included in the Foshan First People's Hospital. The clinical data of patients were collected from hospital records, and the serum level of Slit3 was measured using an enzyme-linked immunosorbent assay. Logistic regression was employed to determine odds ratios for Slit3 and other indicators associated with the development of RA-ILD. Results In total, 232 patients were included. Patients with RA in the high Slit3 subgroup were older, were more likely to be positive for rheumatoid factor and anti-cyclic citrullinated peptide antibody, and had higher levels of erythrocyte sedimentation rate and C-reactive protein. They also showed higher Disease Activity Scores (DAS28-CRP, DAS28-ESR, SDAI) as well as elevated pro-inflammatory cytokine interleukin-6. Notably, logistic regression analysis indicated that the elevated serum level of Slit3 was an independent predictor of RA-ILD. Conclusions Elevated serum level of Slit3 was associated with higher disease activity and served as an independent risk factor of RA-ILD. Slit3 may hold promise as a useful serum biomarker and therapeutic target for RA-ILD.

Importance:A notable proportion of patients with ankylosing spondylitis (ie, radiographic axial spondyloarthritis [r-axSpA]) experience either inadequate responses or intolerance to available therapies, highlighting the need for additional treatment options that offer sustained disease control. Objective:To evaluate the efficacy and safety of vunakizumab, a novel anti-interleukin (IL) 17A monoclonal antibody, in r-axSpA. Design, Setting, and Participants:This randomized, double-blind, placebo-controlled, phase 2 to 3 clinical trial was conducted in 38 hospitals in China. A total of 548 adult patients (aged ≥18 years) with active r-axSpA were enrolled between June 9, 2021, and March 30, 2023. Data were analyzed from November 3, 2023, to January 14, 2024. Interventions:In phase 2, patients were randomized 2:2:1 to receive a subcutaneous dose of vunakizumab, 120 mg, vunakizumab, 240 mg, or placebo; vunakizumab, 120 mg was determined to be the recommended dose after interim analysis. In phase 3, patients were randomized 2:1 to receive vunakizumab, 120 mg or placebo at weeks 0, 2, 4, 8, and 12; from week 16, all patients received vunakizumab, 120 mg every 4 weeks through week 32. Main Outcomes and Measures:The primary end point was the proportion of patients with improvement of 20% or greater in Assessment of Spondyloarthritis International Society response criteria (ASAS20) at week 16. Secondary end points were ASAS20 at week 32, improvement of 40% or greater in ASAS response criteria (ASAS40) at weeks 16 and 32, improvement of 20% or greater in 5 of 6 ASAS domains (ASAS5/6) at weeks 16 and 32, and changes from baseline at weeks 16 and 32 in Bath Ankylosing Spondylitis Disease Activity Index score, Bath Ankylosing Spondylitis Functional Index score, Bath Ankylosing Spondylitis Metrology Index score, 36-Item Short Form Survey score, and Ankylosing Spondylitis Quality of Life Questionnaire score. Results:During the entire study, 548 patients were randomized (mean [SD] age, 33.0 [8.9] years; 440 [80.3%] male), including 294 receiving vunakizumab, 120 mg and 146 receiving placebo. At week 16, the ASAS20 response rate was significantly higher with vunakizumab, 120 mg vs placebo (193 [65.6%] vs 62 [42.5%], respectively; difference, 23.2% [95% CI, 11.8%-34.0%]; P < .001); the ASAS40 response rate also favored vunakizumab, 120 mg vs placebo (136 [46.3%] vs 35 [24.0%], respectively; difference, 22.3% [95% CI, 13.3%-31.3%]; P < .001). Responses with vunakizumab were sustained through 32 weeks. During the placebo-controlled period, incidence of adverse events (246 [83.7%] vs 119 [81.5%]) were comparable in the vunakizumab, 120 mg and placebo groups. Conclusions and Relevance:In this randomized clinical trial of patients with r-axSpA, vunakizumab, 120 mg significantly improved signs and symptoms of r-axSpA at week 16 compared with placebo, with sustained efficacy through 32 weeks, and demonstrated a tolerable safety profile. These findings support vunakizumab, 120 mg as a new treatment option for active r-axSpA. Trial Registration:ClinicalTrials.gov Identifier: NCT04840485.

Implant-associated infections and foreign body responses remain major challenges in orthopedic and biomedical implant applications. In this study, we report a novel strategy to enhance the antibacterial and biocompatibility properties of magnesium (Mg) alloy implants by applying a biodegradable polycaprolactone (PCL) coating embedded with the host-defense peptide, caerin 1.9 (F3). Three Mg-based specimens, including pure Mg, cold-extruded AZ31, and fully annealed AZ31 (3A), were evaluated following PCL-F3 surface modification. The PCL-F3 coatings demonstrated sustained antibacterial efficacy both in vitro and in vivo, effectively inhibiting methicillin-resistant Staphylococcus aureus (MRSA) for up to 168 h. Among the groups, the 3A-PCL-F3 condition exhibited the most notable performance, with substantially enhanced corrosion resistance, reduced inflammatory responses, and no detectable toxicity to vital organs. In vivo proteomic and metabolomic analyses further revealed that the 3A-PCL-F3 implants promoted the expression of osteogenic markers and activated pathways related to bone mineralization and hemostasis, while avoiding prolonged inflammatory activation at 3 months post-implantation. Notably, histological and cytokine ELISA data confirmed favorable tissue responses, including suppressed IL-1β and IL-10 levels and signs of early immune activation that subsided over time. These findings indicate that PCL-F3-coated Mg alloys, particularly the 3A variant, represent a promising solution for biodegradable implants with dual antibacterial and regenerative functionality. This work lays the foundation for developing degradable Mg alloy biomaterials with enhanced biocompatibility and multifunction for clinical use.

Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease accompanied by high incidence rate of interstitial lung disease (ILD). Slit guidance ligand 3 (Slit3) which is a member of axon guidance molecule superfamily, mediates a series of biological activities. In this study, we explored the potential clinical significance of Slit3, especially as a serum biomarker for evaluating RA-ILD. Patients diagnosed with RA were recruited between July 2022 to April 2024 from the Department of Rheumatology of Foshan First People's Hospital. The clinical data were collected from hospital records, serum level of Slit3 was detected by enzyme-linked immunosorbent assay (ELISA). Among 232 recruited patients, RA patients in high Slit3 subgroup were older, showed higher proportion of positive rheumatoid factor (RF) status and anti-cyclic citrullinated peptide antibody (APCA) status, as well as higher levels of ESR, CRP, Disease Activity Score in 28 joints with four variables, including C-reactive protein (DAS28-CRP) or erythrocyte sedimentation rate (DAS28-ESR), Simplified Disease Activity Index (SDAI) and pro-inflammatory cytokine interleukin-6 (IL-6). Notably, Logistic regression analysis showed that elevated serum Slit3 was an independent relevant factor of RA-ILD (OR 1.005 [95% CI 1.002-1.008], P=0.002). Our findings evealed that serum Slit3 was associated with disease activity in RA and may hold promise as a useful serum biomarker and therapeutic target for RA-ILD. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement Yes ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The Ethics Committee of Foshan First People's Hospital approved this study, and all subjects signed informed consent forms before clinical data collection. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All relevant data are within the manuscript and its Supporting Information files.

Rheumatoid arthritis (RA) is a chronic, inflammatory, systemic autoimmune disease characterized by cartilage, bone damage, synovial inflammation, hyperplasia, autoantibody production, and systemic features. To obtain an overall profile of the immune environment in RA patients and its association with clinical features, we performed single-cell transcriptome and T-cell receptor sequencing of mononuclear cells from peripheral blood (PBMC) and synovial fluid (SF) from RA patients, integrated with two large cohorts with bulk RNA sequencing for further validation and investigation. Dendritic cells (DCs) exhibited relatively high functional heterogeneity and tissue specificity in relation to both antigen presentation and proinflammatory functions. Peripheral helper T cells (TPHs) are likely to originate from synovial tissue, undergo activation and exhaustion, and are subsequently released into the peripheral blood. Notably, among all immune cell types, TPHs were found to have the most intense associations with disease activity. In addition, CD8 effector T cells could be clustered into two groups with different cytokine expressions and play distinct roles in RA development. By integrating single-cell data with bulk sequencing from two large cohorts, we identified interactions among TPHs, CD8 cells, CD16 monocytes, and DCs that strongly contribute to the proinflammatory local environment in RA joints. Of note, the swollen 28-joint counts exhibited a more pronounced association with this immune environment compared to other disease activity indexes. The immune environment alternated significantly from PBMCs to SF, which indicated that a series of immune cells was involved in proinflammatory responses in the local joints of RA patients. By integrating single-cell data with two large cohorts, we have uncovered associations between specific immune cell populations and clinical features. This integration provides a rapid and precise methodology for assessing local immune activation, offering valuable insights into the pathophysiological mechanisms at play in RA.

Magnesium (Mg) alloys show outstanding promise for development of degradable implants for hard tissue engineering. However, rapid corrosion and associated reductions in mechanical properties has limited their clinical application. Furthermore, bacterial infections remain an ongoing challenge for implants. Previously, we established that the magnesium alloy, AZ31(Mg-3%Al-1%Zn-0.4%Mn) in a fully annealed form, exhibits improved biocompatibility and corrosion resistance over both pure Mg and cold-extruded AZ31. Multi-omics analyses of tissues of Sprague-Dawley (SD) rats revealed that annealed AZ31 does not significantly activate inflammation and immune responses, while it enhanced signalling in tissue cell proliferation associated pathways. Furthermore, we employed coatings incorporating the host defence peptide (CHDP), caerin 1.9 (abbreviated as F3) into a biocompatible polymer, polycaprolactone (PCL), to develop functional 3-dimensional surface coating to improve biocompatibility and antibacterial performance of the Mg alloy materials. In this study, we have assessed the responses from MC3T3-E1 cells cultured with the Mg alloys to further understand cellular responses. The annealed AZ31 alloy stimulated proliferation of mice osteoblast precursor cells and caused upregulation in expression of Brpf1 protein and other signalling pathways related to bone mineralization and haemostasis, which promote bone tissue formation. The coated and annealed AZ31 alloy (F3-PCL-3A) demonstrated exceptional biocompatibility, causing no adverse effects on hepatic or renal function, and displaying no observable changes in vital organs three months after implantation in SD rats. F3-PCL-3A displayed long-lasting and stable antibacterial properties both in vitro and in vivo . Proteomics and metabolomics analyses of tissues in direct-contact with implants revealed that F3-PCL-3A did not activate inflammation or immune-associated signalling pathways in SD rats 3 months post-implantation. Meanwhile, it activated inflammatory responses, especially phagocytosis pathways up to 72 hours post implantation, indicating enhanced antibacterial capability during the acute stage after implantation. In summary, F3-PCL-3A shows outstanding promise for degradable implants with active antibacterial capabilities for internal fixation and fracture repair.

BackgroundTraumatic brain injury (TBI) is a major cause of neurological disability, and current treatments have limited effectiveness. Recent studies have emphasized the potential of exosomes derived from umbilical cord mesenchymal stem cells (UC-MSCs-Exo) in TBI treatment, but the molecular mechanisms underlying their therapeutic effects are not fully understood.Methods and resultsIn this study, UC-MSCs-Exo was isolated using ultracentrifugation and intraventricularly injected to TBI rat model. The neurofunctional motor function of the rats was evaluated using the modified neurological severity score (mNSS), and the activation of microglia was assessed through immunofluorescence detection of IBA1 expression levels. Additionally, we established an in vitro neuroinflammatory model using BV2 microglia to investigate the effects of UC-MSCs-Exo and miRNA-21. Our findings indicate that UC-MSCs-Exo promote neurological recovery in TBI rats and inhibit excessive microglia activation. Furthermore, UC-MSCs-Exo highly expresses miRNA-21 and inhibited the proliferation, migration, and release of inflammatory mediators of BV2 microglia by transporting miRNA-21.ConclusionsThe present study suggests that the promotion of neurological recovery in TBI rats by UC-MSCs-Exo may be attributed to the inhibition of excessive microglia activation through miRNA-21.

The transcription factor Olig2 is highly expressed throughout oligodendroglial development and is needed for the differentiation of oligodendrocyte progenitor cells (OPCs) into oligodendrocytes and remyelination. Although Olig2 overexpression in OPCs is a possible therapeutic target for enhancing myelin repair in is-chemic stroke, achieving Olig2 overexpression in vivo remains a formidable technological challenge. To address this challenge, we employed lipid nanoparticle (LNP)-mediated delivery of Olig2 synthetically modified messenger RNA (mRNA) as a viable method for in vivo Olih2 protein overexpression. Specifi-cally, we developed CD140a-targeted LNPs loaded with Olig2 mRNA (C-Olig2) to achieve targeted Olig2 protein expression within PDGFR alpha+ OPCs, with the goal of promoting remyelination for ischemic stroke therapy. We show that C-Olig2 promotes the differentiation of PDGFR alpha+ OPCs derived from mouse neural stem cells into mature oligodendrocytes in vitro, suggesting that mRNA-mediated Olig2 overexpression is a rational approach to promote oligodendrocyte differentiation and remyelination. Furthermore, when C-Olig2 was administered to a murine model of ischemic stroke, it led to improvements in blood-brain barrier (BBB) integrity, enhanced remyelination, and rescued learning and cognitive deficits. Our compre-hensive analysis, which included bulk RNA sequencing (RNA-seq) and single-nucleus RNA-seq (snRNA-seq), revealed upregulated biological processes related to learning and memory in the brains of mice treated with C-Olig2 compared to those receiving empty LNPs (Mock). Collectively, our findings highlight the therapeutic potential of multifunctional nanomedicine targeting mRNA expression for ischemic stroke and suggest that this approach holds promise for addressing various brain diseases. Statement of significance While Olig2 overexpression in OPCs represents a promising therapeutic avenue for enhancing remyelina-tion in ischemic stroke, in vivo strategies for achieving Olig2 expression pose considerable technological challenges. The delivery of mRNA via lipid nanoparticles is considered aa viable approach for in vivo pro-tein expression. In this study, we engineered CD140a-targeted LNPs loaded with Olig2 mRNA (C-Olig2) with the aim of achieving specific Olig2 overexpression in mouse OPCs. Our findings demonstrate that C-Olig2 promotes the differentiation of OPCs into oligodendrocytes in vitro, providing evidence that mRNA-mediated Olig2 overexpression is a rational strategy to foster remyelination. Furthermore, the intravenous administration of C-Olig2 into a murine model of ischemic stroke not only improved blood-brain barrier integrity but also enhanced remyelination and mitigated learning and cognitive deficits. These results un-derscore the promising therapeutic potential of multifunctional nanomedicine targeting mRNA expression in the context of ischemic stroke. (c) 2023 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

We read with interest the study by Adam et al reporting a textbooklevel pipeline for biomarker identification in metabolomic datasets. 2 This study pointed to a longstanding issue in metabolomic studies. The lack of robust strategies for biomarker discovery would leave readers with the impression that ‘the biomarker is outofnowhere but works’, especially in those scarcely studied biomarkers. Herein, we provided a strategy that infers the potential of understudied biomarkers for further investigation. A previous study has shown that gut bacterial dysbiosis can cause preeclampsia (PE) symptoms through the gut–placenta axis. However, the role and potential therapeutic effect of gut metabolites as key mediators between bacteria and host remain elusive in PE. Thus, we assessed the faecal metabolome in an agematched case–control study to complete the theory of the gut–placenta axis and identify metabolites with therapeutic potential for PE. For metabolites depleted in PE, the variable importance in projection score indicated that ononetin contributed the most to group separation (figure 1A). However, little is known about ononetin, except for it being a plantbased deoxybenzoin and a potent TRPM3 antagonist, let alone its links with PE. Thus, we designed a twoarms strategy to predict the potential association between ononetin and PE (figure 1B, methods detailed in online supplemental material). The first arm was based on prior knowledge that ononetin and its catabolites inhibit TRPM3 and activate TRPA1, respectively. We obtained their interaction genes from the STRING database (Search Tool for the Retrieval of Interaction Gene/ Proteins, version 11.5) and identified 33 genes, which were enriched in PErelated pathways, including negative regulation of systemic arterial blood pressure and inflammatory response (figure 1C, online supplemental tables S1–S3). The second arm assumed that metabolites correlated with ononetin abundance or shared structural similarity might be involved in similar biological functions. 7 We identified ononetinrelated metabolites in two branches, those that correlated with abundance and those that share structural similarity. In the first analysis, 43 ononetincorrelated metabolites were identified by weighted correlation network analysis (WGCNA), (online supplemental table S4). Then, these metabolites were searched in STITCH (Search Tool for Interactions of Chemicals, version 5.0) and 22 genes were detected, which are involved in vasoconstriction, vasodilation and regulation of systemic arterial blood pressure (figure 1D, online supplemental tables S5 and S6). In the latter branch, we used PubChem to search for structurally similar metabolites with ononetin and selected the top 10 relevant metabolites that interacted with 29 genes that were enriched in vascular endothelial growth factors (VEGF) signalling and regulation of angiogenesis (essential in placental formation and growth) (figure 1E, online supplemental tables S7–S9). Based on the bioinformatic evidence, ononetin was administrated orally into NGnitroLarginine methyl ester (LName) PE mouse model to validate its therapeutic effect. We divided the mice Letter

Chemotherapy-induced premature ovarian insufficiency (CIPOI) triggers gonadotoxicity in women undergoing cancer treatment, leading to loss of ovarian reserves and subfertility, with no effective therapies available. In our study, fecal microbiota transplantation in a cisplatin-induced POI mouse model reveals that a dysbiotic gut microbiome negatively impacts ovarian health in CIPOI. Multi-omics analyses show a significant decrease in Limosilactobacillus reuteri and its catabolite, β-resorcylic acid , in the CIPOI group in comparison to healthy controls. Supplementation with L. reuteri or β-RA mitigates cisplatin-induced hormonal disruptions, morphological damages, and reductions in follicular reserve. Most importantly, β-RA pre-treatment effectively preserves oocyte function, embryonic development, and fetus health, thereby protecting against chemotherapy-induced subfertility. Our results provide evidence that β-RA suppresses the nuclear accumulation of sex-determining region Y-box 7, which in turn reduces Bcl-2-associated X activation and inhibits granulosa cell apoptosis. These findings highlight the therapeutic potential of targeting the gut-ovary axis for fertility preservation in CIPOI.

Developing cancer screening methods, particularly by Surface-enhanced Raman scattering (SERS) of serum samples, is crucial for early cancer diagnosis, given SERS's high biocompatibility and comprehensive spectra fingerprints. However, development of SERS as a cancer diagnostic tool is impeded by the lack of a high-quality serum database and robust, adaptable diagnostic models. In our study, we optimized SERS active Ag-Au nanorods and serum pretreatment to obtain high-quality spectra from 215 women's serum samples and then normalized the spectra to create a reliable serum database. Quadratic discriminant analysis (QDA) of this database could classify cancer patients from healthy individuals with an accuracy of 91.2%. By the kernel density estimation (KDE) algorithm on QDA scores, we calculated the probability distribution function (PDF) for two groups with which the diagnostic accuracy was significantly enhanced, exemplified by a 139/143 accuracy rate at 90% probability decision threshold. Notably, both PDF and QDA models can be refined iteratively with an increase in database increment. Our findings underscore the potential of label-free serum SERS in noninvasive cancer screening, particularly for distinguishing cervical cancer patients from healthy individuals.

BACKGROUND:Tumor heterogeneity plays essential roles in developing cancer therapies, including therapies for breast cancer (BC). In addition, it is also very important to understand the relationships between tumor microenvironments and the systematic immune environment.METHODS:Here, we performed single-cell, VDJ sequencing and spatial transcriptome analyses on tumor and adjacent normal tissue as well as axillar lymph nodes (LNs) and peripheral blood mononuclear cells (PBMCs) from 8 BC patients.RESULTS:We found that myeloid cells exhibited environment-dependent plasticity, where a group of macrophages with both M1 and M2 signatures possessed high tumor specificity spatially and was associated with worse patient survival. Cytotoxic T cells in tumor sites evolved in a separate path from those in the circulatory system. T cell receptor (TCR) repertoires in metastatic LNs showed significant higher consistency with TCRs in tumor than those in nonmetastatic LNs and PBMCs, suggesting the existence of common neo-antigens across metastatic LNs and primary tumor cites. In addition, the immune environment in metastatic LNs had transformed into a tumor-like status, where pro-inflammatory macrophages and exhausted T cells were upregulated, accompanied by a decrease in B cells and neutrophils. Finally, cell interactions showed that cancer-associated fibroblasts (CAFs) contributed most to shaping the immune-suppressive microenvironment, while CD8+ cells were the most signal-responsive cells.CONCLUSIONS:This study revealed the cell structures of both micro- and macroenvironments, revealed how different cells diverged in related contexts as well as their prognostic capacities, and displayed a landscape of cell interactions with spatial information.

Background: Rheumatoid arthritis (RA) is a systemic autoimmune disease causing joint dysfunction. As diseasemodifying anti-rheumatic drugs (DMARDs) have poor efficacy in 20% to 25% of RA patients, additional novel RA medications are urgently needed. Schisandrin (SCH) has multiple therapeutic effects. However, whether SCH is effective against RA remains unknown. Purpose: To investigate how SCH affects the abnormal behaviours of RA fibroblast-like synoviocytes (FLSs) and further elucidate the underlying mechanism of SCH in RA FLSs and collagen-induced arthritis (CIA) mice. Methods: Cell Counting Kit-8 (CCK8) assays were used to characterize cell viability. EdU assays were performed to assess cell proliferation. Annexin V-APC/PI assays were used to determine apoptosis. Transwell chamber assays were used to measure cell migration and invasion in vitro. RT-qPCR was used to assess proinflammatory cytokine and MMP mRNA expression. Western blotting was used to detect protein expression. RNA sequencing was performed to explore the potential downstream targets of SCH. CIA model mice were used to assess the treatment efficacy of SCH in vivo. Results: Treatments with SCH (50, 100, and 200 mu M) inhibited RA FLSs proliferation, migration, invasion, and TNF-a-induced IL-6, IL-8, and CCL2 expression in a dose-dependent manner but did not affect RA FLSs viability or apoptosis. RNA sequencing and Reactome enrichment analysis indicated that SREBF1 might be the downstream target in SCH treatment. Furthermore, knockdown of SREBF1 exerted effects similar to those of SCH in inhibiting RA FLSs proliferation, migration, invasion, and TNF-a-induced expression of IL-6, IL-8, and CCL2. Both SCH treatment and SREBF1 knockdown decreased activation of the PI3K/AKT and NF-kappa B signalling pathways. Moreover, SCH ameliorated joint inflammation and cartilage and bone destruction in CIA model mice. Conclusion: SCH controls the pathogenic behaviours of RA FLSs by targeting SREBF1-mediated activation of the PI3K/AKT and NF-kappa B signalling pathways. Our data suggest that SCH inhibits FLS-mediated synovial inflammation and joint damage and that SCH might have therapeutic potential for RA.

目的 探讨骨膜蛋白(POSTN)与类风湿关节炎(RA)患者疾病活动度的相关性.方法 回顾性分析佛山市第一人民医院风湿免疫科2019年1月-2022年1月收治的114例类风湿关节炎患者,依据DAS28评分将患者分为疾病活动组72例和疾病稳定组42例,同期纳入在本院体检中心体检健康者60名作为健康对照组.比较三组患者的一般临床资料,外周血和关节滑膜组织中POSTN蛋白水平,分析POSTN蛋白与类风湿关节炎患者疾病活动度的相关性,并进一步采用logistic回归模型分析疾病活动期的可能危险因素.结果 类风湿关节炎患者的血清POSTN 蛋白与类风湿因子(r=0.232,P=0.013)、C 反应蛋白(r=0.321,P＜0.05)、病程(r=0.338,P＜0.05)、DAS28评分(r=0.673,P＜0.05)呈显著相关性.类风湿关节炎患者的外周血POSTN蛋白水平显著高于健康对照组,疾病活动组患者外周血的POSTN蛋白水平(7.6±1.4)ng/mL高于疾病稳定组(5.6±1.6)ng/mL和健康对照组(3.2±1.6)ng/mL,差异有统计学意义(F=134.203,P＜0.05),且疾病活动组滑膜组织中POSTN蛋白表达高于疾病稳定组患者[(1.0±0.1)ng/mL vs.(0.6±0.1)ng/mL],差异有统计学意义(P＜0.05).类风湿关节炎患者的外周血POSTN蛋白水平(RR=2.024,95％CI:1.311～3.126)是判断RA疾病活动度的独立危险因素.结论 POSTN蛋白和RA患者的疾病活动度呈显著相关性,可作为独立危险因素预测RA患者的疾病活动度.

ABSTRACT Caerin 1 is a family of host defense peptides with antimicrobial properties originally isolated from the Australian tree frog. Both caerin 1.1 and caerin 1.9 have been demonstrated to inhibit infections caused by multiple antibiotic-resistant bacteria, both in vitro and in vivo. In this current study, we conducted a comparison of the minimum inhibitory concentrations of caerin 1.1 and caerin 1.9 (referred to as caerin 1.1/1.9) with commonly used antibiotics against various bacteria, including Staphylococcus aureus, Copper-Green Pseudomonas aeruginosa, Acinetobacter baumannii, and Streptococcus haemolyticus. Our findings reveal that caerin 1.1/1.9 not only prevent the formation of biofilms by A. baumannii but also exhibit a therapeutic effect on established biofilms. Furthermore, our study demonstrates that caerin 1.1/1.9 significantly inhibit the growth of methicillin-resistant Staphylococcus aureus (MRSA) strains in a murine skin infection model. A quantitative proteomic analysis conducted as part of our study indicates that caerin 1.1/1.9 primarily activate oxidative phosphorylation pathways, along with several pathways associated with tissue repair and growth. These findings were observed in comparison to untreated tissues that were infected with MRSA in mice. In summary, our research outcomes suggest that caerin 1.1/1.9 exhibit the potential to serve as promising drug candidates for treating complicated antibiotic-resistant bacterial infections in humans. IMPORTANCE Caerin 1.1 and caerin 1.9, natural antimicrobial peptides derived from tree frogs, have demonstrated the ability to inhibit the growth of antibiotic-resistant bacteria, comparable to certain widely used antibiotics. Additionally, these peptides exhibit the capacity to prevent or treat biofilms formed by bacteria in conjunction with bodily components. The mechanisms underlying their antibacterial effects were investigated through a mouse model of bacterial skin infection, utilizing proteomic analysis as a technological approach.

免疫组库是指特定时间段内机体内T细胞和B细胞多样性的总和,其多样性特征反映了机体免疫系统对抗各种抗原的免疫应答能力.风湿免疫性疾病以自身抗原导致免疫功能紊乱为特征.通过高通量测序研究免疫组库的多样性是当今一大热点,其在风湿性疾病中的研究亦日益剧增,免疫组库特征不仅为风湿病的发病机制提供了新的见解,还为其临床诊治提供了有用的信息.本文总结了免疫组库多样性在各类风湿病中的研究现状,并强调了免疫组库在风湿免疫性疾病中的应用价值.

Background Cervical cancer (CC) is the 3 rd most common cancer in women and the 4 th leading cause of deaths in gynaecological malignancies, yet the exact progression of CC is inconclusive, mainly due to the high complexity of the changing tumour microenvironment (TME) at different stages of tumorigenesis. Importantly, a detailed comparative single-nucleus transcriptomic analysis of tumour microenvironment (TME) of CC patients at different stages is lacking. Methods In this study, a total of 42,928 and 29,200 nuclei isolated from the tumour tissues of stage-I and II CC patients and subjected to single-nucleus RNA sequencing (snRNA-seq) analysis. The cell heterogeneity and functions were comparatively investigated using bioinformatic tools. In addition, label-free quantitative mass spectrometry based proteomic analysis was carried out. The proteome profiles of stage-I and II CC patients were compared, and an integrative analysis with the snRNA-seq was performed. Results Compared with the stage-I CC (CCI) patients, the immune response relevant signalling pathways were largely suppressed in various immune cells of the stage-II CC (CCII) patients, yet the signalling associated with cell and tissue development was enriched, as well as metabolism for energy production suggested by the upregulation of genes associated with mitochondria. This was consistent with the quantitative proteomic analysis that showed the dominance of proteins promoting cell growth and intercellular matrix development in the TME of CCII group. The interferon-α and γ responses appeared the most activated pathways in many cell populations of the CCI patients. Several collagens, such as COL12A1 , COL5A1 , COL4A1 and COL4A2 , were found significantly upregulated in the CCII group, suggesting their roles in diagnosing CC progression. A novel transcript AC244205.1 was detected as the most upregulated gene in CCII patients, and its possible mechanistic role in CC may be investigated further. Conclusions Our study provides important resources for decoding the progression of CC and set the foundation for developing novel approaches for diagnosing CC and tackling the immunosuppressive TME.

To study clinical effects of joint lesion and liver-kidney function in acute gouty arthritis patients with etoricoxib. Methods:A total of 120 consecutives as acute gouty arthritis from June 2016 to June 2017 were enrolled and divided randomly into control and observation groups, each of 60 cases. The patients in control group received diclofenac sodium 75mg/d and they in observation group adopted etoricoxib 60mg/d, they all got intro-articular corticosteroid injection(compoundbetamethasone3. 5mg+2% lidocaine0. 5ml+NS. 0. 5ml) if necessary, the course was 5d;then to compare differences of joint lesion scores including pain, tenderness, swelling, and skin blush before and after treatment 1d,3d,5d;the levels of serum uric acid, ESR, C reactive protein (CRP) and white blood cell , liver function(ALT), kidney function(serum creatinine)and complications. Results: The joint lesion scores in the two groups were gradually lower,and they in observation group were significantly less(P <0. 05). The levels of uric acid, ESR, CRP and white blood cell after 5d in observation group were significantly lower ( P <0 . 05 ) . The liver-kidney function and complications in the two groups were no statistical differences ( P >0 . 05 ) . Conclusion:It is safe and effective of Etoricoxib for ccute gouty arthritis patients,which can greatly improve joint le-sion without increasing liver-kidney function damages.

Mg alloys have mechanical properties compatible with human bones. However, their rapid degradation and associated foreign body reactions in vivo significantly limit their application for human implants. In this study, three differently processed Mg alloys, pure Mg (PM), cold extruded Mg alloy AZ31 (CE AZ31), and fully annealed AZ31 Mg alloy (FA AZ31) were comparatively investigated for their potential as implants using a rat model. All three implanted Mg alloys do not show any impact on hepato- and renal function, nor any signs of observable changes to vital organs. Proteomics analysis of tissues directly contacting the implants 2.5 months post implantation revealed that FA AZ31 activates very few inflammation and immune associated signaling pathways; while the CE AZ31 and PM produce more significant inflammatory responses as confirmed by cytokine array analyses. Further, FA AZ31 activated pathways for cell organization and development that may improve the recovery of injured tissues. Structurally, EBSD analysis reveals that the FA AZ31 alloy has a higher ratio of first-order pyramidal orientated (10–11) {10–1–2} grain texture with a value of 0.25, while PM and CE AZ31 alloys have lower ratios of first-order pyramidal orientated texture with the values of 0.16 and 0.17, respectively. This is associated with recovery and recrystallisation during annealing which promotes grain texture which exhibits enhanced degradation behaviours and induces a more limited immune response in vivo. In conclusion, the FA AZ31 demonstrated better biocompatibility and corrosion resistance and is a promising candidate for metal-based degradable implants which warrants further investigation.