Background Spinal cord injury (SCI) induces a damaging oxidative microenvironment exacerbating secondary injury. The traditional Chinese medicine (TCM) drug-pair Dangshen and Huangqi, known for antioxidant and neuroprotective effects, yields key components luteolin (Lut) and astragaloside IV (AST), both promising in oxidative stress-related neurological disorders but unexplored in combination for SCI.

Methods We investigated the synergistic antioxidant effects of Lut-AST combination therapy using an in vitro oxidative stress model in PC12 cells, and subsequently assessed its neuroprotective efficacy through behavioral assessments and histopathological analyses in a rat model of severe SCI. Finally, we utilized network pharmacology and molecular docking to predict and explore the potential of the Lut-AST drug pair for treating SCI through multi-target therapy.

Results Our study demonstrated that the Lut-AST drug pair synergistically attenuated oxidative stress-induced cytotoxicity. Lut-AST treatment effectively promoted nerve repair and functional recovery in SCI rats. A significant recovery of motor functions was observed accompanied by reduced accumulation of reactive oxygen species. Neuroinflammation and glial scars were largely alleviated, while the distribution of 5-hydroxytryptamine and neurofilament-positive nerve fibers was evidently increased.

Conclusion These findings confirm Lut-AST’s therapeutic efficacy in mitigating post-SCI oxidative stress and unveil novel insights into traditional Chinese medicine’s inherent multi-component synergistic interactions, suggesting potentiated outcomes through integrated antioxidant mechanisms and multi-target regulation. This study provides a paradigm for optimizing TCM-derived neuroprotective strategies by leveraging component synergy, informing novel combinatorial therapies for SCI management.

Background Trastuzumab deruxtecan (T-DXd) has revolutionized the therapeutic landscape for HER2-positive gastric cancer (GC). However, tumor heterogeneity poses a significant challenge in overcoming T-DXd resistance. This study aimed to delineate the mechanisms underlying primary and acquired resistance to T-DXd in GC.

Methods We performed single-cell RNA sequencing on GC tumor tissues from the DESTINY-Gastric06 study, including treatment-naive baseline samples and those with primary or acquired resistance to T-DXd. Dimensionality reduction and unsupervised clustering were applied to identify distinct cell clusters within the tumor tissues. High-dimensional weighted gene co-expression network analysis was employed to identify key gene modules associated with T-DXd resistance. Cell-cell communication was analyzed using CellChat. Key findings were experimentally validated through multiplex immunofluorescence, immunohistochemistry, and functional assays in cellular models.

Results Weighted gene co-expression network analysis identified the red and purple modules as being strongly correlated with primary and acquired T-DXd resistance, respectively. Notably, MUC3A was upregulated in patients with primary resistance and its overexpression was identified as a potential predictor of shorter progression-free survival in response to T-DXd therapy. Moreover, cystatin C, a gene implicated in linker cleavage, was upregulated during the development of acquired resistance. Tumor microenvironment profiling revealed that T-DXd initially promoted immune-cell infiltration and enhanced antigen presentation. However, with the development of resistance, the tumor microenvironment shifted to an immunosuppressive state, characterized by reactivation of transforming growth factor-beta signaling and upregulation of programmed cell death protein-1 (PD-1).

Conclusion These findings provide novel insights into mechanisms underlying T-DXd resistance and highlight potential therapeutic targets for overcoming T-DXd resistance in GC.

Background Smooth muscle cells (SMCs) exhibit remarkable plasticity, undergoing extensive phenotypic switching to generate a highly heterogeneous population within atherosclerotic plaques. While recent studies have highlighted the contribution of SMC-derived macrophage-like cells to plaque inflammation, the specific molecular drivers governing the transition to these pathogenic states remain poorly understood.

Methods Here, we re-analyzed single-cell RNA sequencing data from lineage-traced mice to dissect SMC heterogeneity during atherogenesis. Trajectory analysis revealed that SMCs transdifferentiate into a distinct pro-inflammatory macrophage-like subpopulation (macrophage 4) via an intermediate “stem-endothelial-monocyte" cell state. Integrated gene regulatory network inference and in silico perturbation modeling identified interferon regulatory factor 7 (IRF7) as a master transcriptional regulator orchestrating this specific pathogenic transition.

Results Clinically, IRF7 expression was significantly upregulated in unstable and advanced human atherosclerotic plaques, correlating strongly with inflammatory macrophage burden. In vivo, ApoE − / − mice challenged with a high-fat diet exhibited robust upregulation of IRF7 in aortic plaques, which co-localized with macrophage markers. Crucially, SMC-specific knock-down of Irf7 using an AAV-SM22 α -shIRF7 vector significantly attenuated atherosclerotic plaque progression, reduced necrotic core formation, and enhanced fibrous cap stability. Mechanistically, Irf7 silencing preserved the contractile SMC phenotype and inhibited the accumulation of pro-inflammatory SMC-derived macrophage-like cells within the lesion.

Conclusions These findings identify IRF7 as a critical checkpoint in maladaptive SMC phenotype switching. We demonstrate that IRF7 drives the transdifferentiation of SMCs into a pro-inflammatory macrophage-like state, thereby fueling plaque instability. Consequently, therapeutic strategies capable of inhibiting IRF7-mediated SMC plasticity may prove effective in stabilizing vulnerable atherosclerotic plaques.

Protein function is inherently spatial: the same molecule can elicit distinct biological outcomes depending on its localization, interacting partners, and surrounding microenvironment. Spatial proteomics enables systematic in situ characterization of protein localization, abundance, and interactions across subcellular to tissue scales, surpassing the resolution and contextual information accessible to conventional bulk proteomics. Recent technological advances including DNA-barcoded multiplexing methods, cyclic fluorescence platforms, and mass spectrometry imaging have substantially increased multiplexing capacity, sensitivity, and spatial accuracy. These capabilities directly support clinically relevant applications, such as tumor immune microenvironment analysis, mapping of protein aggregation in neurodegeneration, growth factor dynamics during tissue repair, patient stratification, pharmacodynamic mapping, and target-engagement assessment. Computational innovations, including graph neural networks, self-supervised embeddings, and workflow management tools (e.g. Snakemake, Nextflow), further enhance cell segmentation, noise reduction, and multi-modal data integration, enabling extraction of robust, spatially resolved proteomic information from complex tissues. Future research will aim to standardize protocols, enable real-time clinical analysis, and develop 3D spatial proteome maps to advance spatial proteomics toward precision diagnostics and targeted therapies.

Objective Previous antibiotic therapy is acknowledged to potentially reduce the efficacy of single-agent immune checkpoint inhibitors. Nevertheless, the impact of antibiotics on the results for patients undergoing chemoimmunotherapy remains unclear. This research investigated the influence of antibiotic treatment on the effectiveness of chemoimmunotherapy in advanced non-small cell lung cancer (NSCLC).

Methods We recorded the characteristics of patients with advanced NSCLC and assessed potential associations between the use of antibiotics and the efficacy of chemoimmunotherapy. A mouse model using Lewis lung carcinoma (LLC) cell lines was developed to assess the effects of antibiotics on the gut microbiome and metabolites. Fecal samples were analyzed using 16S rRNA gene sequencing and ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS) methods. Mouse fecal and serum samples and 16 human stool samples were used to validate the identified differentially metabolites. Deoxycholic acid (DCA) was further applied to a LLC mouse model.

Results This study included 387 NSCLC patients, among whom 86 patients had used antibiotics within the 30 days before the first cycle of chemoimmunotherapy (ATB group), and 301 patients had not used antibiotics (non-ATB group). Notable discrepancies were observed in overall survival and progression-free survival between the two groups, with overall survival recorded at 18.4 months versus 32.0 months, and progression-free survival at 7.6 months versus 13.0 months, in the ATB and non-ATB groups respectively. At the phylum level, the relative abundances of Proteobacteria, Cyanobacteria, and Deinococcus were increased in the ATB mice, while Firmicutes, Bacteroidetes, and Verrucomicrobia were decreased. We detected significant differences in DCA levels in the fecal and serum samples from mice as well as in the fecal sample from humans between the ATB and non-ATB groups. The respective proportions of CD4⁺ and CD8⁺ cells were greater in the non-ATB group than in the ATB group, whereas the proportion of Ki67-positive cells was greater in the ATB group. DCA was applied to LLC mice, and DCA along with chemoimmunotherapy effectively inhibited tumor growth in a LLC mouse model. The expression of programmed cell death ligand 1 increased in the DCA group.

Conclusions Antibiotic exposure is associated with decreased efficacy of chemoimmunotherapy in patients with NSCLC via dysregulation of the gut microbiome and DCA metabolism.

Objectives Gastric-type adenocarcinoma (GAS), an aggressive subtype of non-human papillomavirus (HPV)-associated (NH-PVA) cervical adenocarcinomas (ADC), remains a treatment-refractory disease with poor prognosis. This study aims to explore the oncogenic mechanism and efficacious therapeutic target of GAS.

Methods We included 19 NHPVA and 153 HPVA ADC patients from our center to investigate clinicopathological features. We collected 3 GAS and 2 usual-type endocervical adenocarcinomas (UEA) for single-cell RNA sequencing and T-cell receptor se-quencing. We conducted immunohistochemical staining of 25 GAS and 25 UEA samples and multicolor immunohistochemical staining of 2 GAS samples for validation. We explored the efficacy of anti-clusterin (OGX-011) and/or cisplatin (DDP) for GAS based on GAS-derived tumoroids.

Results Based on clinical data, we clinicopathologically verified the malignancy of GAS. Through single-cell RNA sequencing, we delineated key cell subtypes including GAS epithelial cells, “GAS-enriched fibroblasts”, “GAS-associated γδ T cells”, and CD8 + exhausted T cells enduring heat stress and contributing to GAS aggressive phenotype. Regarding validation, we verified clusterin (CLU)-associated heat stress, highlighted the potential role of CLU-associated stress in promoting immune escape, and established a four-gene signature ( CLU, PDGFB, TIGIT, C3 ) indicating poor prognosis of GAS induced by CLU-associated stress and immune escape. Based on GAS-derived tumoroids retaining the histological features, CLU-associated stress, and genetic profile of parental tumor, we validated the anti-tumor and sensitizing DDP efficacy of targeting CLU.

Conclusion CLU-associated heat stress of key cell subtypes contributed to the malignant GAS microenvironment. Additionally, we pioneeringly constructed GAS-derived tumoroids and suggested that combining CLU-targeted treatment and DDP could improve the therapeutic efficacy for GAS.

Background Minor glomerular abnormalities (MGAs) are histopathologically heterogeneous renal lesions with subtle structural changes and latent clinical manifestations, yet their molecular mechanisms remain poorly characterized and underexplored.

Methods In this study, we employed pressure cycling technology-assisted sample preparation combined with data-independent acquisition mass spectrometry to systematically compare the proteomic profiles of distant non-neoplastic tissues ( n = 24) and MGA tissues ( n = 27).

Results A total of 9 529 protein groups were quantified with a false discovery rate < 1%, and 1 338 differentially expressed protein groups were identified (fold-change > 2 or < 0.5, P < 0.05), including 190 downregulated and 1 148 upregulated protein groups in MGA tissues. Gene ontology analysis revealed that the downregulated proteins were enriched in cell adhesion, ion binding, and molecular transport, whereas the upregulated proteins were enriched in transcriptional regulation, DNA replication/repair, and nucleic acid binding. Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated inhibition of metabolic pathways and the peroxisome proliferator-activated receptor signaling pathway, as well as the activation of basal transcription factors and nucleotide excision repair in MGAs. Further screening revealed 13 core upregulated nuclear proteins (e.g. YY1, TAF9, RFC1, and POLR1D) with a > 90% detection rate in MGA tissues; these proteins are functionally associated with renal inflammation, cell proliferation, and the DNA damage response.

Conclusion Our study establishes a high-resolution proteomic landscape of MGAs, provides novel insights into their molecular pathogenesis, and identifies potential tissue biomarkers and therapeutic targets. The pressure cycling technology-assisted data-independent acquisition workflow also offers a robust technical framework for proteomic analysis of microscale renal biopsy samples.

Chimeric antigen receptor (CAR)-engineered immune cells have revolutionized cancer immunotherapy, expanding from the established success of CAR-T cells to a diverse array of cellular platforms. While seven Food and Drug Administration-approved CAR-T cell products demonstrate unprecedented efficacy in hematologic malignancies, significant limitations persist, including severe inflammatory toxicities, resistance in solid tumors, and manufacturing barriers. These challenges have catalyzed extensive research to extend CAR engineering into alternative effector cell types, such as unconventional T cell subsets, natural killer (NK) cells, macrophages, neutrophils, and dendritic cells, as well as non-immune platforms. Each cell type exhibits distinct antitumor mechanisms, persistence profiles, safety characteristics, and manufacturing requirements, positioning them to address complementary therapeutic needs. This review provides a comprehensive overview of diverse CAR-engineered cellular platforms, encompassing their biological properties, advantages, sourcing strategies, and manufacturing processes, alongside current clinical progress and optimization approaches. Beyond oncology, these platforms have demonstrated significant potential in treating autoimmune diseases, infections, cardiac fibrosis, and senescence-associated disorders. By leveraging distinct immune and non-immune cell types to mediate cytotoxicity or suppress pathogenic cells, CAR technology provides versatile therapeutic avenues across varied disease contexts. Through synthesis of recent advances in CAR platform diversity, this review identifies opportunities for targeted optimization and explores future directions for broadening CAR-based therapeutic applications.