Background: Cervical cancer (CC) remains a significant global health challenge despite advancements in screening, HPV vaccination, and therapeutic strategies. Tumor heterogeneity, driven by epigenetic modifications, affects immune evasion, metastasis, and treatment response. Cancer-associated fibroblasts (CAFs) play a crucial role in CC progression and therapy resistance. Single-cell sequencing offers new insights but remains underutilized in CC research. This study integrates single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and deconvolution analysis to identify key genes and immunotherapy targets. By constructing a prognostic model and exploring the immune microenvironment, we aim to provide novel insights into CC pathogenesis and potential therapeutic strategies. Methods: We utilized scRNA-seq, spatial transcriptomics, deconvolution analysis, and pseudotime trajectory mapping to delineate fibroblast subtypes within the tumor immune microenvironment (TIME) of CC. Functional annotations, differential gene expression profiling, cell–cell communication pathways, and transcription factor networks were systematically analyzed. A prognostic model based on bulk RNA-seq data was constructed and validated through survival analysis, with correlations to immune microenvironment characteristics. Functional experiments investigated the role of SDC1, a critical mediator of fibroblast-tumor crosstalk. Additionally, Fibroblast–tumor cell co-culture systems and functional assays were employed to investigate the paracrine role of SDC1. The CAF MYH11⁺ subpopulation was isolated via fluorescence-activated cell sorting (FACS). Multiplex immunofluorescence and immunohistochemical analyses were performed on both cultured cells and human cervical cancer tissue samples to characterize the spatial distribution and dynamic remodeling of MYH11 during stromal reorganization. Results: Six distinct fibroblast subtypes were identified, including the C0 MYH11 + fibroblasts, which exhibited unique roles in stemness maintenance, metabolic activity, and immune regulation. Spatial and functional analyses revealed that the C0 subtype is central to tumor-fibroblast interactions, particularly through the MDK-SDC1 signaling axis. The prognostic model incorporating fibroblast-specific markers demonstrated robust predictive power for patient survival outcomes. Additionally, in vitro SDC1 knockdown significantly inhibited CC cell proliferation, migration, and invasion. Fibroblasts show spatially regulated heterogeneity, with activation markers enriched in the tumor zone and MYH11 highest in normal zones, indicating dynamic stromal remodeling. C0 MYH11 + CAF Promotes Tumor Cell Proliferation, Migration, and Inhibits Apoptosis via Soluble SDC1. Conclusion: Our results illustrate, in some ways, the possible immunomodulatory and tumor supporting roles of CAFs in CC TIME and highlight the possibility that the MDK-SDC1 pathway is a promising therapeutic target. This study not only promotes a partially new understanding of temporal heterogeneity in CC, but also provides a possible reference base for the development of new biomarkers and immunotherapy approaches to improve clinical outcomes.

Glioma, particularly glioblastoma, is a highly aggressive brain tumor with poor prognosis and limited treatment options. Recent research highlights the role of MANF (Mesencephalic Astrocyte Derived Neurotrophic Factor) in tumor biology, yet its specific mechanisms in glioma remain underexplored. This study aims to elucidate the role of MANF in glioma and its underlying mechanisms of action. We conducted bioinformatics analysis using TCGA data to identify MANF-related pathways, followed by cellular assays and subcutaneous tumor models for functional validation. Experiments included Western blot and qRT-PCR analysis to investigate the effects of MANF on glioma cell proliferation, migration, and stemness gene expression. MANF was found to be highly expressed in tumor tissues and associated with poor prognosis in glioma patients. Endogenous MANF regulates tumor cells by modulating the TGF-β/SMAD4/p38 pathway, promoting stemness and enhancing malignant behaviors, including migration and invasion. Exogenous MANF, however, did not significantly affect stemness gene expression but contributed to glioma cell proliferation. MANF emerges as a promising therapeutic target for glioma. This study clarifies MANF's specific mechanisms, offering insights into its potential for targeted glioma therapies.

Background: The most aggressive type of skin cancer, melanoma, has a high prevalence of metastases and a poor prognosis. Despite advancements in treatment, drug resistance and tumor microenvironment heterogeneity, especially involving neuro-immune interactions, continue to exist. The goal of this study is to uncover the cellular heterogeneity of melanoma in order to pinpoint molecular targets and tumor-promoting subtypes. Methods: Melanoma single-cell RNA sequencing data came from GEO. Twelve cell types were discovered after Harmony batch effect adjustment and Seurat quality control; melanoma cells were subtyped. Functional expression of differential genes was examined using gene ontology and gene set enrichment. Cytotrace measured subtype differentiation potency. PySCENIC revealed transcription factor regulatory networks, and CellChat predicted intercellular communication between malignant cell subtypes and other cell types. Functional experiments with A375 and MEWo cell lines—lentiviral shRNA knockdown, CCK-8 proliferation, wound healing, transwell migration, and flow cytometry apoptotic assays—confirmed the analysis. Results: Single-cell RNA sequencing was utilized to analyze melanoma cell subtypes and their interactions in the tumor microenvironment. C2 IGFBP3 +, which had the lowest CytoTRACE2 score and was enriched in late tumor stages, affected melanoma development. This subtype expressed prominent immunomodulatory pathways. The C2 subtype mediate signaling to fibroblasts/pericytes via the PROS pathway and myeloid/plasmacytoid dendritic cells via the MHC-II pathway. The C2 subtype was strongly linked with FOSL1 expression, which may affect gene transcription and illness progression. FOSL1 knockdown significantly increased apoptosis and decreased melanoma cell motility and proliferation in vitro . Conclusion: We identified immunoregulatory C2 IGFBP3 + melanoma cell subtypes in our investigation, and FOSL1 was a crucial transcription factor that aided in cell migration, proliferation, and survival. Because the C2 subtype involves the MHC-II and PROS signaling pathways, which have been shown to have roles in neuroimmunology, neuroinflammation, and pain regulation, it may serve as a hub for neuro-immune interactions in the tumor microenvironment. Precision treatments for melanoma may be advanced by focusing on the FOSL1 or C2 subtype pathways, which may assist in overcoming immunotherapy resistance.

Background: Traumatic brain injury (TBI) is increasingly recognized as a systemic inflammatory disorder, with neutrophils playing a critical role in secondary injury. However, the phenotypic heterogeneity and clinical significance of neutrophil subsets in the early TBI immune landscape remain unclear, limiting their utility in nursing prognostic assessment and individualized care planning.Methods: We performed an integrated multi-omics analysis—combining single-cell RNA sequencing (scRNA-seq), bulk transcriptomics, and proteomics—to dissect neutrophil diversity post-TBI. A distinct SLFN4⁺ neutrophil population was identified and further validated through in vitro functional assays and serum profiling in a TBI patient cohort. Clinical correlations and nursing stratification models were constructed to evaluate prognostic relevance.Results: At 24 hours post-injury, scRNA-seq revealed four neutrophil clusters in mouse brains. Among these, the SLFN4⁺ subset exhibited N1-like polarization, pro-inflammatory activation, and metabolic rewiring favoring glycolysis and oxidative phosphorylation. Regulon and pseudotime analyses highlighted its transitional regulatory potential. SLFN4⁺ neutrophils actively engaged in TNF and CCL-mediated communication with monocytes/macrophages. Functionally, silencing SLFN4 or STAT2 enhanced neutrophil proliferation and reduced inflammatory cytokine secretion. Clinically, elevated serum SLFN4 levels in TBI patients were associated with poor neurological outcomes and, when incorporated into a composite nursing risk model, significantly improved early prognostic accuracy.Conclusions: SLFN4⁺ neutrophils represent a key inflammatory effector population contributing to early immune dysregulation after TBI. Beyond mechanistic insights, SLFN4 serves as a promising serum biomarker to enhance clinical decision-making and nursing risk stratification. These findings support the integration of immunological biomarkers into precision nursing frameworks to guide early interventions and improve neurorehabilitation outcomes.