Reproductive aging in mice leads to estropause, characterized by estrous cycle irregularity and eventual cessation, yet its underlying mechanism remains unclear. Here, we present a comprehensive single-cell atlas of mouse ovaries across precisely defined reproductive stages—from young (regular cycling) through the estropausal transition (regular vs. irregular cycling) to post-estropause (acyclic)—and of ovary-specific senescent cells defined by high senescence-associated β-galactosidase activity. We mapped transcriptomic dynamics of ovarian aging and characterized the molecular features of ovarian senescent cells. Our analyses revealed that during the estropausal transition, irregularly cycling ovaries exhibited accelerated aging and cellular senescence features compared with regularly cycling counterparts, including increased transcriptional noise, altered conserved aging pathways such as oxidative phosphorylation and proteostasis, hormone dysregulation in granulosa cells, and elevated expression of the senescence marker Cdkn1a and senescence-associated secretory phenotype factors. This atlas delineates the cellular and molecular hallmarks of mouse ovarian aging and ovary-specific senescent cells, providing a resource for understanding the mechanisms underlying the estropausal transition.

Leaf spots caused by Curvularia lunata infection pose a significant threat to global maize production. Although resistance gene breeding faces challenges due to pathogen evolution, the plant microbiome has emerged as a key modulator of disease resistance. However, the mechanisms via which plant genes regulate phyllosphere metabolites to recruit beneficial microbes remain poorly understood. Here, we combined gene mapping, metabolomics, microbiome analyses, cytological analysis, and in vitro and in vivo experiments to investigate the disease resistance mechanism of ZmHPATR1. We first identified that the loss-of-function mutation in ZmHPATR1 significantly increased the levels of fumaric acid, folic acid, and tetrahydrofolic acid in the leaves, leading to the enrichment of the genus Sphingomonas. We further demonstrated that the extracellular polysaccharide, welan gum, biosynthesized by Sphingomonas, effectively inhibited C. lunata growth and disrupted its cell structure. These results enable us to comprehensively understand the complicated mechanisms of plant resistance to disease through a four-level regulatory network that links plant genes, metabolites, microbes, and pathogens. Our findings provide new strategies for targeted microbiome-based disease-resistant breeding and the development of novel biopesticides for maize.

Ovarian cancer is one of the deadliest gynecological malignancies, mainly because of its silent development and the lack of effective conventional methods for diagnosis. Because of their extraordinary stability in bio fluids and expression signatures that specifically correlate with tumors, circulating microRNAs (miRNAs) have received much attention as non-invasive diagnostic biomarkers. This meta-analysis evaluates the diagnostic value of circulating miRNAs for early ovarian cancer. A systematic literature review was performed in line with PRISMA-DTA guidelines in PubMed, EMBASE, and Web of Science until March 8, 2025. A total of 24 studies were eligible. Pooled diagnostic metrics (sensitivity and specificity, likelihood ratios, and diagnostic odds ratio (DOR)) were calculated based upon the bivariate random effect model. Summary receiver operating characteristic (sROC) curves were generated and subgroup and meta-regression analyses performed to investigate heterogeneity. Sensitivity analyses were performed to assess the strength of the pooled estimates. The combined sensitivity, specificity and DOR was 0.749 (95

Bacterial therapy represents a promising approach for cancer treatment, with the probiotic Escherichia coli Nissle 1917 (EcN) being a particularly attractive candidate due to its inherent tumor-targeting capability and established safety profile. However, the underlying mechanisms of its tumor-colonization and anti-tumor activity remain unclear. Lactate, a key immunosuppressive metabolite generated abundantly in tumors via the Warburg effect, critically shapes the tumor microenvironment (TME) and promotes immune evasion. Given EcN's innate capacity to metabolize lactate, we hypothesize that lactate utilization is pivotal for its therapeutic activity and could be enhanced to potentiate anti-tumor immunity. To test this, we first constructed a lactate-utilization-deficient EcN strain (EcN-ΔLA) via CRISPR-Cas9, which exhibited severely impaired tumor colonization and abrogated anti-tumor effects, establishing lactate metabolism as a non-redundant mechanism for EcN's anti-tumor function. We then engineered a stable, plasmid-free strain with enhanced lactate-scavenging capacity (BELAC) by genomically integrating the lactate transporter gene lldP and dehydrogenase lldD to achieve hyper-scavenging of tumor-derived lactate. BELAC demonstrated superior lactate scavenging and robust tumor suppression across multiple syngeneic mouse models accompanied by increased infiltration of CD8+ T cells and significant remodeling of the immunosuppressive TME. Notably, in the cold tumor 4T1 breast cancer model, BELAC colonized tumors and reduced lactate levels but failed to elicit significant anti-tumor activity as a monotherapy, underscoring the limitations of lactate depletion alone in some low-immunogenic tumors. Critically, BELAC preserved EcN's tumor-targeting specificity and safety while maintaining high genomic stability. Collectively, this work establishes lactate metabolism as a fundamental requirement for EcN's anti-tumor activity and presents BELAC as a clinically translatable microbial therapeutic that exploits lactate depletion to reprogram the TME and enhances anti-tumor immunity in immunoresponsive contexts.

The precise enhancement of nutritional quality in silage maize is a core strategy for increasing livestock production efficiency. Through evolutionary analysis of multiple plant species, we identified two functionally synergistic upstream open reading frames (uORF1 and uORF2) within the 5' untranslated region of the gene encoding GDP-L-galactose phosphorylase (GGP). By leveraging a natural translation-enhancing haplotype of uORF1 (Hap2) and performing CRISPR/Cas9-mediated targeted mutagenesis of the highly conserved uORF2, we successfully engineered an elite dual-uORFs variant. This variant significantly increased vitamin C (Vc) content and concurrently improved key silage quality traits, including crude protein and phosphorus levels, without inducing growth penalties. Transcriptomic profiling further elucidated the molecular mechanisms by which the dual-uORFs variation coordinately regulates Vc biosynthesis and the improvement of silage quality. Our findings deepen the understanding of the conventional paradigm of single-uORF regulation and provide a novel strategy and superior germplasm resources for the precision breeding of high-Vc, high-quality silage maize.

Excessive fertilization poses a major threat to sustainable agriculture, resulting in resource waste and environmental degradation. The ecological composite fertilizer (ECF) combined with fertilizer reduction represents a promising strategy to improve rhizosphere microbial diversity in wheat systems. A field experiment, containing six treatments, namely traditional compound fertilizer (TF, applied at the conventional rate) with a 10% reduction (TF90), TF90 plus ECF application (TF90+ECF), TF with a 15% reduction (TF85), TF85 plus ECF application (TF85+ECF), TF with a 20% reduction (TF80), and TF80 plus ECF application (TF80+ECF), was conducted to explore the influences of fertilizer reduction combined with ECF application on wheat yield and rhizosphere soil microbial diversity. Results showed that the TF85+ECF treatment achieved the highest wheat yield at 8,717.33 kg ha−1, which was significantly greater than all other treatments and represented a 30.63% increase over the TF85 treatment. The TF85+ECF group significantly enhanced the activities of the carbon and nitrogen cycling enzymes β-1, 4-glucosidase glucosidase (BG) and urease (UE), and increased the abundances of the functional genes cbbLR and amoA. In the +ECF treatment groups (TF90+ECF, TF85+ECF, and TF80+ECF), linear discriminant analysis effect size (LEfSe) and specialization-occupancy (SPEC-OCCU) analyses identified keystone microbial taxa, including positively correlated taxa with biocontrol and metabolic versatility (e.g., Trichoderma, Solicoccozyma) and negatively correlated potential pathogens (e.g., Alternaria). Co-occurrence network analysis revealed that the TF85+ECF group streamlined bacterial network architecture while enhanced fungal network complexity and connectivity. Mantel tests and correlation analyses indicated that soil organic carbon, BG activity, and cbbLR gene abundance were significantly linked to microbial community structure, and keystone taxa were strongly correlated with soil nutrient cycling functions. Our findings provide a microbiome-based strategy and a novel perspective for sustainable wheat production and targeted microbial management in agriculture.

Purpose: This study aimed to develop, characterize, and clinically evaluate a novel 6% nitroglycerin (NTG) Aloe vera gel formulation as a transdermal alternative to conventional sublingual NTG therapy in patients with stable angina pectoris. Patients and Methods: A randomized, parallel-group, double-blind clinical trial was conducted involving 150 patients with stable angina, allocated to receive either 6% NTG-Aloe vera gel or standard sublingual NTG tablets (0.6mg). The gel was assessed for physicochemical parameters, FTIR compatibility, stability under ICH guidelines, and in vitro drug release kinetics. Dermatological safety was evaluated via skin irritation testing. Clinical efficacy was determined by Numeric Pain Rating Scale (NPRS), chest tightness scores, Seattle Angina Questionnaire (SAQ), and vital parameters, including blood pressure and oxygen saturation. Results: The NTG Aloe vera gel demonstrated desirable physicochemical properties, sustained zero-order drug release, and excellent stability with no significant degradation. Clinical data revealed that the gel provided gradual, sustained pain and chest tightness relief, superior SAQ scores (p< 0.001), and improved tolerability compared to sublingual NTG. Importantly, the gel induced a controlled hypotensive response without abrupt cardiovascular changes, indicating enhanced safety. No dermatological adverse effects were reported. Conclusion: This pilot study showed 6% NTG-Aloe vera gel offers a stable, effective, and patient-friendly transdermal delivery system for angina management, meriting further large-scale and long-term evaluations. Registration: Clinical Trial Registry-India (CTRI/2024/11/076947).

Wastewater-based epidemiology (WBE) has strong potential for community-level environmental exposure surveillance, but current applications remain limited in chemical scope and rarely integrate biomarker discovery with targeted quantification. Here, we developed an integrated wastewater exposomics framework combining untargeted Phase II metabolite profiling with targeted measurement of established volatile organic compound (VOC) biomarkers to characterize community-level chemical exposure. Across sixteen sewersheds in Louisville Metro, Jefferson County, Kentucky, including eleven neighborhood catchments and five centralized treatment facilities-we detected 194 Phase II metabolites derived from 145 parent xenobiotics, including mercapturic acids, glucuronides, and sulfates associated with aldehydes, aromatic hydrocarbons, epoxides, and related environmental chemicals. Several metabolites were consistently detected across sites, whereas many exhibited marked spatial heterogeneity, with greater variability at the neighborhood scale. Targeted liquid chromatography-mass spectrometry analysis of VOC biomarkers similarly revealed greater variability in neighborhood catchments than in treatment-center influent. Together, these findings demonstrate that integrating untargeted and targeted approaches extends WBE beyond limited target panels, enabling broader biomarker coverage and improved spatial resolution of community-level chemical exposures. Further refinement of normalization strategies, metabolite libraries, and data integration will enhance its utility for population-level exposure assessment.

The ovary is the first organ to age in the human body, affecting both fertility and overall health. However, the biological mechanisms underlying human ovarian aging remain poorly understood. Here we present a comprehensive single-nuclei multi-omics atlas of four young (ages 23-29 years) and four reproductively aged (ages 49-54 years) human ovaries. Our analyses reveal coordinated changes in transcriptomes and chromatin accessibilities across cell types in the ovary during aging, notably mTOR signaling being a prominent ovary-specific aging pathway. Cell-type-specific regulatory networks reveal enhanced activity of the transcription factor CEBPD across cell types in the aged ovary. Integration of our multi-omics data with genetic variants associated with age at natural menopause demonstrates a global impact of functional variants on gene regulatory networks across ovarian cell types. We nominate functional non-coding regulatory variants, their target genes and ovarian cell types and regulatory mechanisms. This atlas provides a valuable resource for understanding the cellular, molecular and genetic basis of human ovarian aging. The molecular and cellular mechanisms underlying ovarian aging are incompletely understood. Here the authors provide single-nuclei RNA and ATAC-seq of human ovarian tissue from four young and four reproductively aged donors, revealing coordinated transcriptomic and epigenomic changes across cell types and highlighting a role for mTOR signaling in reproductive aging.

In single-cell transcriptomic analysis, accurate cell type annotation forms the essential basis for all downstream analysis and data interpretation. While time consuming manual annotation requires extensive prior knowledge, available automated cell annotation tools lack a unified, curated human cell reference to ensure successful identification of all cell types present in any given dataset. To fill in the gap we create “CellMap” a comprehensive single cell gene expression reference database of known cell types across most human tissues by compiling, curating and integrating single cell datasets from multiple sources including Human Protein Atlas, Tabula Sapiens, ArrayExpress and GEO. CellMap contains 151 different celltypes/states including 94 epithelial, 31 brain, 4 from soft tissues, 8 stromal and 15 major immune celltypes from 42 human tissues. Projecting pseudo-bulk profiles of the 151 celltypes into 2-dimensional UMAP revealed major clusters of cells from the nervous system, respiratory system, gastrointestinal tract, proximal digestive system, female tissues and endocrine tissues. Immune cells and stromal cells are generally clustered together regardless of their tissue origin. We also curated about 200 immune cell phenotypes from the ImmGen database for fine-tuning annotation of immune cell subpopulations. Importantly by combining CellMap and the SingleR algorithm, we successfully identified the cell lineage of the tumor and minor cell types originated from the biopsy site of a breast cancer metastatic sample; the latter were left unannotated with reference derived from breast tissue or misclassified using Human Primary Cell Atlas. In addition, CellMap allowed us to identify highly specific tissue markers and cell type markers. Based on 1241 identified marker genes, we trained a random forest model to predict cancer types using 80% of the TCGA tumor samples of each cancer type (n = 7160) and tested on a 20% hold-out testing set (n = 1774), achieving a median of 95.4% accuracy across cancer types. Cancer types with low accuracy include esophageal carcinoma (mostly mis-classified as head and neck squamous cell carcinoma) and rectum adenocarcinoma (mostly mis-classified as colon adenocarcinoma). The misclassification could be explained by the high similarity between these cancer types in terms of cell-of-origin. Finally a webportal for exploring celltypes in CellMap and an open-source R-package for automated cell annotation of single cell data and cell lineage prediction for bulk RNA-seq data were developed. This work provides valuable resources for conducting single-cell analysis and cancer cell-of-origin analysis. Additionally, it helps identify closely related cell types across tissue types, which is particularly useful since diseases originating from related cell types may share similar therapeutic targets. Yuping Zhang, Gabriel Cruz, Hanbyul Cho, Chandan Kumar-Sinha, Rahul Mannan, Jin Chen, Xuhong Cao, Saravana M. Dhanasekaran, Arul M. Chinnaiyan. CellMap: a comprehensive human single cell gene expression reference for automated cell annotation and cancer cell-of-origin analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1063.

Although immune checkpoint inhibitors specifically targeting the PD-1/PD-L1 axis have exhibited remarkable clinical success, they are not uniformly effective across all patient cohorts. Immunotoxins, a novel class of cancer therapeutics, offering a promising alternative. PD-L1, which is also present in certain normal tissues, limits its suitability as an ideal target for immunotoxins. The probiotic strain of E. coli Nissle 1917 (EcN) could target and colonize to solid tumors, which positions it as a promising candidate for tumor tissue-specific delivery of anti-tumor proteins. In this study, we constructed a PD-L1-targeted immunotoxin, designated as αPD-L1-PE38, by fusing an anti-PD-L1 nanobody and a clinically validated PE38 toxin. This immunotoxin exhibited potent cytotoxic activity against tumor cells while showed slightly cytotoxic activity against normal cells. To effectively deliver the αPD-L1-PE38 to tumor tissues, we engineered the EcN strain to release the immunotoxin induced by L-arabinose. Upon induction, the immunotoxin was efficiently secreted, and exhibited robust anti-tumor activity mainly by inducing cell apoptosis both in vitro and in vivo. Furthermore, we enhanced the immunotoxin's affinity for PD-L1 by optimizing the linker between the nanobody and PE38 toxin. The engineered EcN expressing the optimized immunotoxin, achieved superior anti-tumor activity. Collectively, our study suggests that the delivery of immunotoxins through live bacteria to improve safety and efficacy is a promising option in cancer therapeutics.

Introduction:Jatropha mollissima (Pohl.) Baill is a traditional medicinal plant reputed for its hepatoprotective and nephroprotective properties. However, its potential cardioprotective and anti-inflammatory effects, both in vitro and in vivo, remain underexplored. Aim of the study:This study conducted a series of in vitro, in vivo, and ex vivo experiments to determine the cardioprotective properties and anti-inflammatory effect of the aqueous-methanolic leaf extract of J. mollissima. Doxorubicin-induced cardiotoxicity, thrombolytic, anticoagulant, antioxidant, vasorelaxant, anti-inflammatory, and calcium channel-blocking activities were determined. Materials and Methods:The study involves a phytochemical evaluation, along with HPLC analysis. The antioxidant activities of the J. mollissima extract were determined using in vitro assays, including DPPH, SOD, NO, and H2O2. In vitro and in vivo anticoagulants, antithrombolytic agents, vasorelaxants, and biochemical assays were performed to determine Jm's protective effect. Cardiac inflammatory markers (TNF-α, IL-1β, IL-6, and IL-10) were evaluated via real-time PCR. Doxorubicin was used as the positive control. Results:In an in-vitro anticoagulant experiment, J. mollissima displayed a substantial increase in activated partial thromboplastin, prothrombin, and clotting time in a dose-dependent manner (20%, 10%, and 5% dilutions) compared with heparin (250 IU/mg) and distilled water. While in-vivo anticoagulant experiment showed a substantial increment in clotting time, prothrombin time, bleeding time, and activated partial thromboplastin time in a dose-dependent manner (25 mg/kg, 50 mg/kg, and 100 mg/kg) in rats after 1-week of treatment in comparison with heparin (50 IU/mg) and distilled water. For the thrombolytic (in vivo and in vitro) experiments, dose-dependent (20%, 10%, and 5% dilutions) significant (p < 0.05) clot lysis was observed compared to streptokinase (30,000 IU) and distilled water. For antioxidant activity, doxorubicin (intraperitoneally at 10 mg/kg at 0 days) was given, blood samples were extracted (at 21st day) to determine cardiac damage by measuring DPPH, SOD, NO, CK-MB, LDH, Troponin I, serum sodium, and serum potassium in which aqueous-methanolic extract in a dose-dependent manner (600 and 400 mg/kg dilutions) displayed significant (p < 0.005-0.000) decrease in serum level. The cardiac weight-to-body weight ratio showed significant resistance to necrosis caused by the doxorubicin-induced toxic group. HPLC analysis revealed the presence of gallic acid, mandelic acid, quercetin, pyrogallol, and rutin. Gene expression analysis revealed that Jm reduced proinflammatory cytokines (TNF-α, IL-1β, and IL-6) and upregulated the anti-inflammatory cytokine IL-10, with effects comparable to those of doxorubicin. Conclusion:Thus, the anticoagulant, antioxidant, cardioprotective, anti-inflammatory, and thrombolytic properties of J. mollissima are attributed to the presence of various phytochemical constituents, which may act on multiple factors. Its beneficial actions are attributed to the modulation of oxidative stress and neuroinflammatory pathways, suggesting its therapeutic potential in managing cardiotoxicity and other complications.

Ras Association Domain Family 1 (RASSF1) proteins are key modulators of tumor suppression, apoptosis, and cell cycle regulation. Among its isoforms, RASSF1A is the most extensively studied due to its crucial role in maintaining cellular homeostasis and preventing cancer progression. It primarily functions as a scaffold protein, orchestrating critical signaling pathways such as Ras, Hippo, and p53 to regulate cell proliferation and apoptosis. Epigenetic silencing of RASSF1A, predominantly through promoter hypermethylation, is a hallmark of numerous human cancers, including those of the lung, breast, and liver. Structural studies have identified conserved domains within RASSF1A, such as the Ras-association (RA) and SARAH domains, which mediate interactions with Ras-GTP and MST kinases to modulate tumor suppressor pathways. Additionally, post-translational modifications (PTMs), such as phosphorylation, influence RASSF1A stability and activity. Dysregulation of RASSF1A impairs several cellular processes, leading to unchecked cell proliferation and tumorigenesis. Understanding the molecular mechanisms regulating RASSF1A-both epigenetically and post-translationally-provides critical insights into potential therapeutic strategies aimed at restoring its tumor-suppressive functions. This review highlights current knowledge of RASSF1A's role in tumorigenesis and explores emerging approaches to target its dysregulation in cancer therapy.

BACKGROUND:Non-small cell lung cancer (NSCLC), the predominant histological subtype of lung cancer, continues to pose a significant global health challenge due to its elevated incidence and mortality. Traditional Chinese medicine (TCM) has garnered growing recognition for its therapeutic potential in oncology. Croton tiglium L., a classical TCM herb, contains bioactive alkaloids with established anti-tumor properties. Although crotonoside-containing formulations exhibit anticancer efficacy in vivo, the molecular mechanisms underlying its effects on NSCLC are not fully understood. OBJECTIVE:This study aims to delineate the molecular mechanism through which crotonoside exerts therapeutic effects against NSCLC, with particular emphasis on EGFR signaling pathway modulation and associated downstream cascades. METHODS:This study used a multidisciplinary technical system to investigate the mechanism of crotonoside on NSCLC. First, the cytotoxicity of crotonoside on tumor cells was determined through in vitro cell experiments, and its impact on the apoptosis and migration of NSCLC cells was evaluated. Then, a subcutaneous xenograft mouse model was established to verify the anti-tumor effect of crotonoside in vivo. Network pharmacology and transcriptome analysis were applied to explore potential molecular targets. Differential gene expression and pathway enrichment were validated through qRT-PCR, Western Blotting, and immunohistochemistry. RESULTS:Crotonoside significantly inhibited the proliferation, migration, and angiogenesis of NSCLC cells. Mechanistically, it suppressed EGFR activation and attenuated downstream PI3K/Akt and MAPK/ERK signaling pathways, resulting in decreased expression of oncogenic mediators. CONCLUSION:Crotonoside exerts potent anti-NSCLC activity associated with modulation of the EGFR signaling pathway. These findings suggest that crotonoside may serve as a promising lead compound for further structural optimization and development of novel EGFR-targeted or multi-targeted therapeutics against NSCLC.

Background:Ferroptosis, a form of cell death discovered in recent years, is expected to provide new targets for the diagnosis and treatment of hepatocellular carcinoma (HCC) through further research. Methods:Based on data from The Cancer Genome Atlas (TCGA), we screened HCC-associated genes from 259 candidate genes in the FerrDb database. The screened genes were subjected to differential expression analysis, survival analysis, correlation analysis with clinical data, and univariate and multivariate Cox regression analysis. The results were validated with the Gene Expression Profiling Interactive Analysis 2 (GEPIA2) database and the Human Protein Atlas (HPA) database, and signaling pathways were analyzed with the Gene Set Enrichment Analysis (GSEA) enrichment analysis. Human normal hepatocytes and different liver cancer cell lines were used to verify the expression levels of genes, using quantitative reverse transcription PCR (RT-qPCR). Results:Eight ferroptosis-related genes were finally selected, including ACSL3, ASNS, CHMP5, MYB, PCK2, PGD, SLC38A1, and YY1AP1. The expression of eight genes except PCK2 was significantly correlated with a lower survival rate of HCC, and the expression of PCK2 showed a correlation with a higher survival rate of HCC. The expression of all eight genes was also correlated with clinical traits. GSEA enrichment analysis obtained many pathways such as apoptosis, endocytosis, pathways in cancer, Wnt signaling pathway, primary bile acid biosynthesis, and fatty acid metabolism pathway. Conclusion:The ACSL3, ASNS, CHMP5, MYB, PCK2, PGD, SLC38A1, and YY1AP1 genes may become markers and new targets for early diagnosis and prognostic assessment of HCC.

Since recombinant biotherapeutics were accepted in the therapeutic drug market, one of the significant challenges has been the insufficient circulatory half-life of many protein and peptide-based drugs. To overcome this pharmacokinetic limitation, serum albumin has been widely utilized as a molecular carrier due to its intrinsic FcRn-mediated recycling mechanism. Besides human drugs, there is also a significant need for extended half-lives of pet or veterinary drug therapeutics, making the development of albumin-binding domains with cross-species reactivity necessary. VNARs represent the antigen-binding domains of Immunoglobulin new antigen receptors (IgNARs) found in cartilaginous fish (e.g. sharks), which exhibit unique structural properties and hold significant potential for clinical diagnosis and treatment. In this study, we constructed a phage display library by immunizing the white-spotted bamboo shark with human serum albumin (HSA). Through multiple rounds of panning using both human and mouse serum albumins, VNAR-4A12 was successfully screened. Affinity tests verified that VNAR-4A12 exhibited cross-reactivity to multiple serum albumin homologues. Additionally, the structural basis for this cross-reactivity was confirmed, and it was demonstrated that 4A12-sfGFP exhibited prolonged exposure compared to sfGFP in vivo. This multispecies targeting strategy addresses critical limitations in both human and veterinary pharmaceutical development, offering new opportunities for long-term therapy.

Cellular senescence is a complex biological process that plays a pathophysiological role in aging and age-related diseases. The biological understanding of senescence at the cellular and tissue levels remains incomplete due to the lack of specific biomarkers as well as the relative rarity of senescent cells, their phenotypic heterogeneity and dynamic features. This Review provides a comprehensive overview of multiomic approaches for the characterization and biological understanding of cellular senescence. The technical capability and challenges of each approach are discussed, and practical guidelines are provided for selecting tools for identifying, characterizing and spatially mapping senescent cells. The importance of computational analyses in multiomics research, including senescent cell identification, signature detection and interactions of senescent cells with microenvironments, is highlighted. Moreover, tissue-specific case studies and experimental design considerations for individual organs are presented. Finally, future directions and the potential impact of multiomic approaches on the biological understanding of cellular senescence are discussed.

>The addition of biochar (BC) or Arbuscular mycorrhizal fung(AMF) alone has been reported to promote plant growth,while their synergistic effects on Allium schoenoprasum root morphology and rhizosphere fungal community in barren soil is still unclear In this study,we investigated the effects of BC and AMF (Funne liformis mosseae) on plant growth and root morphology in barren soil and revealed the structure of soil fungal communitie Therefore,a greenhouse pot trial consisting of five treatment was enforced.The results showed that the combination of bio char and AMF significantly improved plant biomass,nutrien uptake,mycorrhizal colonization rates and soil properties and significantly impacted rhizosphere fungal community composi tion and structure.Biochar significantly increased the funga community stability and enhanced their positive correlation with plants.Our findings indicated that the combination of AMF and biochar play synergic role for plant growth and rhizosphere fungal community in barren soil.

The symbiotic relationship between arbuscular mycorrhizal fungi (AMF) and plants is well known for its benefits in enhancing plant growth and stress resistance. Research on whether key components of the AMF colonization process, such as MyC factors, can be directly utilized to activate plant symbiotic pathways and key functional gene expression is still lacking. In this paper, we found that, using a hydroponics system with Lotus japonicus, MyC factor analogue chitin oligomer 5 (CO5) had a more pronounced growth-promoting effect compared to symbiosis with AMF at the optimal concentration. Additionally, CO5 significantly enhanced the resistance of Lotus japonicus to various environmental stresses. The addition of CO5 activated symbiosis, nutrient absorption, and stress-related signaling pathways, like AMF symbiosis, and CO5 also activated a higher and more extensive gene expression profile compared to AMF colonization. Overall, the study demonstrated that the addition of MyC factor analogue CO5, by activating relevant pathways, had a superior effect on promoting plant growth and enhancing stress resistance compared to colonization by AMF. These findings suggest that utilizing MyC factor analogues like CO5 could be a promising alternative to traditional AMF colonization methods in enhancing plant growth and stress tolerance in agriculture.

Female reproductive aging affects fertility and overall health. Functional decline in mouse ovary during reproductive aging is accompanied by estrous cycle prolongation and cessation. However, the molecular mechanism underlying reproductive aging concomitant with such cycle changes remains unclear. Using single-cell transcriptomics, we characterized aging signatures in mouse ovaries across the reproductive lifespan ranging from reproductively young (regular cycle) through peri-estropause (regular vs. irregular cycles) to post-estropause age (acyclic). Reproductive aging significantly remodeled cell compositions and increased transcriptional heterogeneity, with more pronounced changes post-estropause, exhibiting coordinated alterations across cell types in the ovary. Genes undergoing monotonic changes during reproductive aging across cell types were consistently enriched in the conserved pathways of aging, including oxidative phosphorylation, stress responses, and proteostasis. Additionally, cell type-specific changes were identified including dysregulation of hormone synthesis in granulosa cells, alterations in collagen and hyaluronan metabolism in stromal and early theca cells, and functional decline of a unique phagocytosis-associated macrophage. Aging also led to a significant decrease in cell-cell communications, particularly between stromal and granulosa cells, and an increase in extracellular vesicle secretion. Furthermore, we found increased expression of the senescence marker Cdkn1a and senescence-associated secretory phenotype (SASP) factors during ovarian aging, especially in granulosa cells. Notably, most of these aging-associated changes were more pronounced in irregular cycling ovaries compared to the regular cycling counterparts at the same age during the peri-estropause stage, suggesting that aging-related molecular changes in the ovary drive the estropausal transition in mice. ### Competing Interest Statement The authors have declared no competing interest.