As an important tissue of the body(accounting for approximately 40％of body weight),skel-etal muscle is composed of various cell types such as muscle fibers,muscle stem cells,and endothelial cells.It participates in physiological processes including movement,energy metabolism,and internal en-vironment homeostasis through temporal and spatial specific regulation.Its development is divided into two critical stages:embryonic and postnatal periods.Abnormal development can lead to diseases such as muscular dystrophy and directly affect the yield and quality of livestock meat.In recent years,the combi-nation of single-cell transcriptomics(scRNA-seq)and spatial omics(single-cell spatial omics technolo-gy)has provided a high-resolution research tool for analyzing the spatiotemporal dynamic regulatory net-work and intercellular interactions in skeletal muscle development.This article reviews the molecular mechanisms of skeletal muscle development and its application value in animal husbandry breeding,and systematically combs the research progress,analysis processes,data resources,and future directions of single-cell omics,spatial omics,and single-cell spatial omics technology in skeletal muscle development.Among them,single-cell omics can reveal the heterogeneity of skeletal muscle cells,myofiber differentia-tion trajectories in different livestock and poultry(such as cattle,pigs,and Tibetan chickens)through methods like pseudotime analysis and RNA velocity analysis.Furthermore,single-cell omics can identify key transcription factors(e.g.,MYF5,MYOD1)and cell communication pathways(e.g.,FGF7-FG-FR2),and simultaneously clarify the molecular differences in myoblast differentiation timing and cell composition ratio among different breeds.Relying on technologies such as Visium and Seq-Scope,spatial omics realizes the spatial localization of gene expression in pathological models of mice,Atlantic salmon,and broiler chickens.Spatial omics also clarifies the spatial distribution laws of neuromuscular junction region-specific genes and inflammation-fibrosis cascade reactions,and makes up for the defect of losing spatial context in single-cell technology.Although there are limited direct application cases of single-cell spatial omics technology,it has already analyzed the abnormal fate of myoblasts in facioscapulohumeral muscular dystrophy through MERFISH technology.In terms of technology selection,it is necessary to consider research objectives,molecular modalities,and resolution requirements.At the same time,data analysis needs to address challenges such as data sparsity through methods like DCA denoising and RCTD cell type mapping.In addition,this article summarizes 16 muscle development-related databases inclu-ding HCA and PanglaoDB.This review further discusses the potential applications of these three types of technologies in the directional regulation of myoblast fate,precise intervention in the growth cycle,im-provement of microenvironment interactions,and the development of multi-omics genetic breeding mod-els.This paper is providing a more comprehensive and detailed theoretical reference and technical sup-port for basic research on skeletal muscle development and practical applications in the animal husbandry industry.

Folate receptor α(FRα),encoded by the FOLR1 gene,is overexpressed in various solid tumors but minimally expressed in normal cells,making it a prime target for anti-tumor therapy.Antibod-y-drug conjugates(ADCs)can accurately deliver cytotoxic agents to tumor cells,enhancing treatment specificity and efficacy.Recent years have seen remarkable progress in the development of FRα-targeted ADCs,with multiple drugs entering clinical trials and demonstrating promising anti-tumor activity and safety.This review covers the latest clinical advances in FRα-targeted ADCs.It provides a detailed in-troduction to the structure and function of FRα,its expression in solid tumors,and the clinical progress of FRα-targeted ADCs.It particularly focuses on the clinical trials of drugs like Mirvetuximab Sor-avtansine,Luveltamab Tazevibulin,and Farletuzumab Ecteribulin.These drugs have shown significant anti-tumor effects in different cancer types,and Mirvetuximab Soravtansine has received FDA accelerated approval.Despite their potential,FRα ADCs face challenges in clinical applications,such as toxicity management,biomarker definition,and clinical translation.Future research should optimize FRα ADC design,including antibody selection,linker stability,and drug load efficiency,and explore their combi-nation with other therapies to enhance efficacy and broaden the therapeutic window.Additionally,in-depth studies on the role of FRα in the tumor microenvironment and its interaction with other signaling pathways will strengthen the theoretical basis for FRα ADC drug development.In summary,the develop-ment of FRα ADC drugs is rapidly advancing and is expected to offer new treatment options for FRα-posi-tive cancer patients.

This study aims to investigate the effects of aerobic exercise on neuroinflammation in AD mice and explore the mechanisms of neuroinflammation regulated by the blood-brain barrier,lipopolysaccharide(LPS)displacement,and glial cell activation.Twenty 3-month-old male APP/PS1 double transgenic mice were used,which were randomly divided into a sedentary group(SE-AD)and an aerobic exercise group(Run-AD),and 10 3-month-old male C57BL/6 mice were used as the control group(WT).The Run-AD group underwent 12 weeks of aerobic training.The results of the water maze showed that aerobic exercise improved the learning and memory capacity of AD mice(P＜0.05).The results of H&E stai-ning and Nissl staining showed that aerobic exercise reduced necrotic cells and inflammatory cell infiltra-tion in the cerebral cortex,as well as nuclear condensation in the CA1 and GD regions of the hippocam-pus(P＜0.05,P＜0.01),and increased the area of Nissl bodies in the cerebral cortex and hippocam-pal CA3 and DG regions.Western blotting and ELISA results showed that aerobic exercise increased the expression of Occludin,ZO-1 and Claudin-5 proteins in the brain(P＜0.01),and decreased the levels of LPS in the brain(P＜0.01).The qRT-PCR results exhibited that aerobic exercise decreased the ex-pression of TLR4,MyD88,NF-κB,IL-1β,and TNF-α mRNA(P＜0.05,P＜0.01).The results of immunofluorescence staining revealed that aerobic exercise reduced the fluorescence area of brain IL-1βand TNF-α proteins(P＜0.05,P＜0.01),as well as the fluorescence area of Iba-1,GFAP,and TLR4 proteins in the cerebral cortex and hippocampus(P＜0.05,P＜0.01).There was a high degree of overlap between Iba-1 and TLR4 fluorescence in the cerebral cortex,and GFAP was localized around Iba-1.In summary,aerobic exercise attenuates neuroinflammation in AD mice by protecting the blood-brain barrier,reducing the displacement of LPS,and subsequently weakening the immune activation of microglia to regulate the TLR4/MyD88/NF-κB signaling pathway to alleviate neuroinflammation.

This review aims to summarize the progress of multi-omics technologies in the study of antibi-otic resistance in Cronobacter,with the goal of gaining a deep understanding of its resistance mechanisms and providing a scientific basis for the development of new treatment methods and prevention strategies.By integrating genomics,transcriptomics,proteomics,and metabolomics,the study analyzes gene varia-tions,expression patterns,protein function changes,and metabolic pathway adjustments in Cronobacter.This includes the use of whole-genome sequencing to reveal gene variations related to antibiotic resist-ance,RNA-seq technology to monitor changes in gene expression patterns,proteomics to study protein expression and function,and metabolomics to analyze dynamic changes in metabolites.The research has found that factors such as biofilm formation and outer membrane proteins significantly affect the antibiotic resistance of Cronobacter.In addition,new potential influencing factors have been identified,including the expression changes of multidrug efflux pump genes,which may play a key role in enhancing antibiotic efflux and reducing intracellular antibiotic concentrations.Multi-omics technologies provide a comprehen-sive and in-depth perspective for the study of antibiotic resistance in Cronobacter,revealing multiple fac-tors and potential mechanisms that affect resistance.Although some new influencing factors have been i-dentified,their specific molecular mechanisms still require further investigation.The application pros-pects of multi-omics technologies are broad,and they are expected to provide important support for the development of new treatment methods and prevention strategies.

Single-cell and spatial omics technologies are spearheading a profound paradigm shift in the life sciences,moving beyond'population averages'to'single-cell resolution'and reintegrating'cellu-lar constitution'with'tissue spatial architecture',thereby dramatically advancing our understanding of biological complexity.This review provides a brief comprehensive overview of recent advancements in the field.Technologically,the evolution has progressed from single-cell transcriptomics to integrated approa-ches capturing multiple molecular layers simultaneously,while the emergence of transcriptome-badsed spatial omics has successfully preserved the spatial positioning of cells or microscosystem within native tis-sues,and further enabling spatial epigenomics and spatial-multiomics.Computationally,artificial intelli-gence and machine learning have become central engines,powering tools for data integration,spatial de-convolution,cellular communication and other novel foundation models,which not only tackle the chal-lenges of massive datasets but also serve as instruments for novel biological discovery.These technological leaps have fostered significant theoretical innovations.In clinical translation,these technologies,particu-larly in precision oncology,demonstrate transformative potential by dissecting tumor heterogeneity,map-ping the spatial architecture of the tumor immune microenvironment,and enabling disease modeling through single-cell-guided deconvolution of bulk data,offering new avenues for diagnosis,prognosis,and personalized therapy.Despite ongoing challenges in technological throughput,computational scalability,and clinical integration,the continued convergence of single-cell and spatial omics with AI promises to propel basic research towards a more mechanistic and predictive era,ultimately reshaping the future of precision medicine.

Single-cell assay for transposase-accessible chromatin sequencing(scATAC-seq)is a power-ful technique for studying cellular heterogeneity and gene regulatory networks,widely applied in epigenet-ic research.However,the complexity of data analysis workflows and high programming requirements have limited its broader adoption among non-programmer researchers.To address this issue,we developed Sig-nac.UIO,a modular and visual scATAC-seq analysis platform based on the R Shiny framework,integra-ting mainstream tools such as Signac and Seurat.The platform includes ten key modules covering quality control,cell filtering,dimensionality reduction,clustering,differential analysis,cell annotation,path-way enrichment,motif analysis,and transcription factor footprinting.Through a graphical user interface,users can perform full analyses and obtain interactive visualization results.The platform's stability and u-tility have been validated using a public PBMC dataset and it is currently deployed online(https://xula-bgdpu.org.cn/Signac.UIO),providing an efficient and user-friendly tool for single-cell epigenomics re-search.

Microglia activation-mediated neuroinflammatory responses serve as a critical pathological ba-sis for the development and progression of various brain diseases.The role of circular RNAs(circRNAs)in the regulation of neuroinflammation is increasingly being recognized.This study aimed to investigate the effect and molecular mechanisms of targeted inhibition of circular RNA Homeodomain Interacting Pro-tein Kinase 3(HIPK3)(circHIPK3)on lipopolysaccharide(LPS)-induced microglial polarization in BV2 cells.The results showed that LPS stimulation significantly induced polarization of BV2 cells towards the pro-inflammatory M1 phenotype and upregulated circHIPK3 expression(P＜0.01).Engineered extra-cellular vesicles(EVs)with rabies viral glycoprotein(RVG)loaded with circHIPK3 siRNA(RVG-EVs-sicHIPK3)were successfully constructed.Transmission electron microscopy(TEM)revealed their typi-cal EV morphology.nanoparticle tracking analysis(NTA)indicated a peak particle size of 70 nmn.And Western blotting analysis confirmed the expression of characteristic membrane marker proteins.Treatment with RVG-EVs-sicHIPK3 significantly suppressed the LPS-induced elevation of inflammatory cytokines(TNF-α,IL-6,IL-1β)in the supernatant and reduced the expression of M1 phenotypic marker proteins(CD16 and CD86)(P＜0.01).Concurrently,RVG-EVs-sicHIPK3 increased the number of mitophago-somes within cells,upregulated the ratio of the autophagy-related proteins LC3-Ⅱ/LC3-I(P＜0.01),and downregulated the expression of the autophagy-related protein p62 and mitochondrial-specific proteins(TOMM20 and TIMM23)(P＜0.01).The mitophagy inhibitor Mdivi-1 significantly reversed the RVG-EVs-sicHIPK3-mediated downregulation of inflammatory cytokine levels,M1 marker proteins,and mito-chondrial protein expression(P＜0.01).This study demonstrates that inhibiting circHIPK3 reduces LPS-induced microglial polarization towards the M1 phenotype.The protective mechanism is closely associated with enhanced mitophagic flux and the promotion of damaged mitochondrial clearance.

Ischemic stroke(IS)research has faced bottlenecks due to the limitations of conventional technologies in resolving cellular heterogeneity and spatiotemporal dynamics.The development of single-cell and spatial omics technologies provides revolutionary tools to break through these constraints.Single-cell omics technologies,by performing high-throughput sequencing on thousands of cells,reveal the high heterogeneity and dynamic state transitions of neurons,glial cells,immune cells,and others post-IS.For instance,microglia contain pro-inflammatory and anti-inflammatory functional subsets,while astrocytes exhibit distinct activation state spectra.Pseudotime analysis further reconstructs the fate trajectories of cells during the damage and repair processes.Spatial omics technologies,conversely,reconstruct spatial maps of gene expression through in situ capture,elucidating molecular gradients between the ischemic core,penumbra,and healthy brain regions,and enabling the analysis of critical cell-cell interaction net-works.Integrating the deep phenotyping capability of single-cell sequencing with the in situ localization information from spatial omics constitutes the current core strategy.This multimodal analysis allows for precise anchoring of cell subtypes to their spatial microenvironments,revealing their distribution patterns and functions,and constructing a more accurate atlas of cell-cell communication.This significantly ad-vances the refined dissection of IS mechanisms.This strategy has already accelerated the discovery of po-tential biomarkers and spatiotemporally specific therapeutic targets.Although challenges remain in sample preparation,data integration,and technical noise,future interdisciplinary collaboration,multi-omics in-tegration,and in-depth mining with artificial intelligence promise to comprehensively transform our under-standing of IS.Ultimately,it holds the potential to promote advances in its early diagnosis,precise sub-typing,and the development of individualized treatment strategies.

Mesenchymal stem cells(MSCs)hold great promise in regenerative medicine due to their multi-lineage differentiation potential and immunomodulatory properties.However,their functional heter-ogeneity and strong dependency on the microenvironment remain major challenges for clinical application.In recent years,the combination of single-cell transcriptome sequencing(scRNA-seq)and spatial tran-scriptomics sequencing(ST-seq)has provided revolutionary tools for systematically deciphering the heter-ogeneity,functional diversity,and microenvironmental interactions of MSCs.Using scRNA-seq,re-searchers have successfully resolved the functional heterogeneity of MSCs and identified key functional subpopulations,such as pro-angiogenic,immunoregulatory,and matrix-remodeling subsets.Meanwhile,ST-seq has revealed the distinct spatial distribution of MSCs within tissues and their dynamic interaction networks with the microenvironment.The integration of these two technologies has not only enabled the construction of a three-dimensional"identity-location-function"atlas of MSCs,but also uncovered spatio-temporal dynamic regulatory mechanisms of specific subpopulations during tissue repair.Looking forward,the combination of ultra-high-resolution ST-seq platforms such as Xenium,multi-omics integration,and artificial intelligence-driven analysis will shift MSCs research from descriptive studies toward precise inter-vention,offering new strategies for functional subpopulation screening and microenvironment reprogram-ming therapy.This review systematically summarizes the latest advances in scRNA-seq and ST-seq tech-nologies in MSC research,discusses their applications in elucidating cellular heterogeneity,spatial micro-environment,and clinical translation,and provides a theoretical basis and technical guidance for preci-sion treatment in regenerative medicine.

Cell death is classified into programmed cell death(PCD)and non-programmed cell death(NCD).Necroptosis is a form of PCD that does not rely on caspases and is regulated by four signaling pathways:receptor-interacting protein kinase 1/3(RIPK1-RIPK3),TIR-domain-containing adapter-in-ducing interferon-β(TRIF)-RIPK3,Z-DNA binding protein 1(ZBP1)-RIPK3,and type Ⅰ/Ⅱ interferon receptors(IFNRs).These pathways interact to regulate the activity of core molecules such as RIPK1,RIPK3,and mixed lineage kinase domain-like protein(MLKL),thereby determining the occurrence of necroptosis.The dysregulation of these pathways can lead to the development of various diseases,inclu-ding cancer.Necroptosis not only inhibits tumor occurrence and progression by promoting tumor cell death,but also creates a tumor microenvironment(TME)conducive to tumor cell growth through its pro-inflammatory properties,thereby promoting tumor growth and metastasis.Therefore,the dual role of nec-roptosis in cancer makes it an important research direction in tumor treatment.This article reviews the key signaling pathways of necroptosis,explores its interactions with other cell death pathways such as cell survival,apoptosis,and pyroptosis.Meanwhile,it analyzes the dual regulatory mechanisms of necropto-sis in cancer progression and discusses the issue of overcoming tumor treatment resistance by modulating necroptosis.It further explores its potential therapeutic targets and application prospects,aiming to pro-vide new intervention strategies and theoretical basis for cancer treatment.