Ginseng's prolonged development renders it susceptible to environmental stresses. Late embryogenesis abundant (LEA) proteins are essential for plant resistance to abiotic stress. Our previous study demonstrated that PgLEA2-50, a member of the LEA protein family, plays a significant role in stress resistance. In this study, we employed IP-MS, bioinformatics, and molecular interaction assays to investigate the mechanisms underlying its stress resistance. PgLEA2-50 formed complex networks with multiple interacting proteins, which were enriched in stress-related processes such as gibberellin (GA) signal transduction, saponin biosynthesis, and the oxidative stress response. Transcriptome analysis revealed that its interacting targets exhibited significant responses to abiotic stress at the transcriptional level. An investigation of the DELLA protein PgRGA4 showed that it was down-regulated following GA induction, with its transcriptional activity inhibited under stress conditions. PgRGA4 was found to be localized in both the nucleus and cytoplasm, and co-immunoprecipitation (CO-IP) confirmed its interaction with PgLEA2-50, suggesting that PgLEA2-50 indirectly regulates GA-mediated stress resistance. This study provides a ginseng-specific case for the role of LEA proteins in stress resistance and identifies a novel gene target for molecular breeding in medicinal plants.

Approximately 20% of China’s land area is desertified or highly desertifiable, where loose sandy soil and low nutrient availability restrict plant growth. Microbial inoculants, as an emerging ecological restoration technology, play a key role in plant growth and soil nutrient activation in sandy regions. However, a systematic understanding of functional differences among microorganisms isolated from different stressed environments remains insufficient. Nine functional microbial strains from three stressed habitats, including sandy land, coastal saline-alkali soil, and heavy metal mining areas, were selected to conduct a three-month pot experiment, investigating their effects on soil nutrient activation, plant growth and microbial communities. Results showed that all inoculants increase plant biomass (by 4.15~25.59%), with KS-33, KS-36, SD-13 and SD-3 significantly promoting biomass in different plant parts (p < 0.05), and with YJ-15 remarkably enhancing root growth (root length increased by 70.83%, p < 0.01). Inoculation reduced bacterial Chao1 by 27.18~53.97%, but increased fungal Chao1 by 12.77~28.38% (except SD-30). Bacterial generalist species proportion increased from 61.12% to 83.78~93.99% after inoculation, higher than the variation degree of the fungal community. Mantel analysis revealed a reverse trend between soil nutrients, water content and plant growth. This may be associated with the increased consumption by plants and microorganisms. In summary, microbial inoculants enhance nutrient cycling processes and plant growth by reshaping soil microbial communities. Performance of microbial inoculants is more likely governed by their inherent ecological functions rather than being entirely determined by their original environments. Despite varying mechanisms, these inoculants can effectively enhance sandy soil microbial communities, providing a theoretical basis for regional ecological restoration.

Introduction Natural polysaccharides exhibit promising pharmacological potential in functional foods. However, their structural heterogeneity and limited bioactivity hinder further applications.Methods In this study, a polysaccharide from blackened jujube pomace was selenylated via the conventional HNO3-Na2SeO3 route, with systematic optimization of reaction parameters to improve controllability, yielding selenium-enriched polysaccharides (BJPP-Se).Results and Discussion Multimodal characterization confirmed successful selenylation, with a reduction in molecular weight from 88.26 kDa to 74.32 kDa and an increase in crystallinity. Density functional theory calculations identified two distinct Se (IV) coordination modes, with one involving Se (IV) as a bridging atom linking two monosaccharide units and the other restricting Se (IV) coordination to a single monosaccharide unit. BJPP-Se has much stronger immunomodulatory performance than native polysaccharides in cyclophosphamide suppressed immunosuppressed mice. Selenium-induced structural reorganization is a very good example of how to design bioactive polysaccharides. This work supports the potential for valorizing polysaccharides from black jujube pomace; however, large-scale techno-economic feasibility and cost-benefit assessment are beyond the scope of this study and warrant future work with clearly defined boundaries and engineering-scale datasets.

Natural killer (NK) cell-based therapies are under assessment for the treatment of various cancers due to their intrinsic ability to distinguish between malignant and healthy cells in an allogeneic context, enabling off-the-shelf manufacturing possibilities. However, cryopreservation reduces both the recovery and function of NK cells, thereby limiting their therapeutic feasibility. In this study, we evaluated three cryoprotectants (CryoStor 10; ZKCELL FM-01; FBS + DMSO) for the cryopreservation of NK cells. Post-thaw viability, ATP levels, and cytotoxicity were assessed and found to have persistent differences between cryopreserved and fresh cells. Transmission electron microscopy, flow cytometry, and Western blot analysis revealed a complex mode of cell death in cryopreserved cells, which could be partially mitigated by adding some death inhibitors. We further investigated the effects of centrifugation on thawed cells, identifying lysosomal stability as a key determinant of cell death. Pretreatment with low-dose LLOMe prior to cryopreservation induced stress granule formation, stabilizing lysosomes and improving cell recovery rates without compromising effector functional capacity. These findings offer new insights for optimizing NK cell cryopreservation and facilitating their clinical application.

Natural killer cells exhibit significant potential within current immunotherapeutic modalities. Hypothermic preservation is a critical step for the clinical transportation and temporary storage of NK cell products. However, during this process, both cell viability and function significantly decrease. The core damage mechanisms remain unclear, and effective, specialized preservation protocols are currently lacking. This study aimed to elucidate the primary modes of cell death and maintain NK cell potency during hypothermic preservation. This study first evaluated the effects of different intravenous fluids and temperatures on NK cell preservation efficiency. Subsequently, various cell death inhibitors, molecular biomarker assays, and electron microscopy techniques were utilized to systematically elucidate the modes of cell death. Based on these findings, we optimized the best-performing fluids by replacing sodium lactate with glucose to enhance energy supply, adding the antioxidant α-tocopherol to mitigate oxidative stress, and elevating the potassium ion concentration (to 22.5 mM) to suppress excessive nutrient uptake and associated oxidative damage. These optimizations were incorporated into a new formulation designated as the GAK solution. The preservation efficacy was comprehensively assessed through in vitro cell viability and cytotoxicity assays, as well as an in vivo leukemia xenograft model. Storage at 4 °C was much better than at 25 °C. Among the tested intravenous fluids, lactated Ringer’s demonstrated the highest efficacy in preserving NK cell viability and function. NK cells undergo complex death pathways during hypothermic preservation, specifically ferroptosis, pyroptosis, and necroptosis. Our optimized GAK effectively mitigates energy depletion and oxidative stress, significantly maintaining the viability and potent cytotoxic capacity of NK cells against various tumor cells after hypothermic storage. In vivo experiments further demonstrated that NK cells preserved in the GAK solution maintained significant antitumor activity, which contributed to prolonged survival in tumor-bearing mice. This study revealed multiple death mechanisms of NK cells during hypothermia and successfully developed a well-defined and highly efficient GAK solution. The GAK solution presents a new approach to the hypothermic preservation of NK cells, facilitating the efficient and convenient transport of NK cell products, which has significant practical implications for the accelerated clinical adoption of cellular immunotherapy.

Microbial contamination in food necessitates effective antimicrobial packaging. While cellulose-based packaging materials suffer from limited antimicrobial efficacy, lack of active functionality, and susceptibility to inducing microbial resistance. To address these challenges, this study synthesized a cationic porphyrin-based covalent organic framework (Por-ICOF) as a multimodal photosensitizer. Por-ICOF was uniformly dispersed via non-covalent interaction within hydroxypropyl methylcellulose (HPMC), creating an HPMC/Por-ICOF composite film. This integration enhanced mechanical strength (increased by 26%), hydrophobicity (WCA 71°), and gas barrier properties (OP reduced by 42%, WVP reduced by 36%). Under visible light, the HPMC/Por ICOF film superior absorption generated reactive oxygen species (ROS) and photothermal effects, inactivating 99.2% of Escherichia coli and 99.95% of Staphylococcus aureus within 20 min. The composite film exhibited excellent biocompatibility and effectively extended the shelf life of strawberries. This cationic modification strategy for cellulose-based films offers a novel avenue for the design of high-performance antimicrobial food packaging materials.

Emerging contaminants pose novel food safety challenges, necessitating highly selective and sensitive detection methods. Molecularly imprinted polymers (MIPs) have been employed extensively in food safety assays due to their predetermined architecture, specific recognition capabilities, practicality, and stability. This review systematically evaluates the structural and performance advantages of MIP-based sensors in targeting foodborne pollutants, emphasizing their role in enhancing analytical selectivity and sensitivity. The article further highlights recent breakthroughs in MIP applications across four critical EC categories: persistent organic pollutants, endocrine disruptors, pharmaceutical and personal-care products, and antibiotic residues. Finally, the current challenges impeding rapid development are identified and future prospects are outlined. Furthermore, the multifaceted technical, economic, and regulatory obstacles to transitioning MIP-based technologies from laboratory prototypes to practical monitoring tools are explored. This systematic review advances the establishment of innovative MIPs-driven emerging contaminants monitoring systems, thereby strengthening food safety protocols and public health safeguards.

BACKGROUND:Radiation-induced colitis (RIC) is a common complication following radiotherapy for pelvic and abdominal malignancies. Dysregulated polarisation of macrophages towards the pro-inflammatory classically activated M1 phenotype is a key driver of RIC progression. PURPOSE:This study sought to examine the protective influence of compound Kushen injection (CKI) against RIC and clarify the mechanisms underlying these protective effects. METHODS AND RESULTS:Mice were subjected to whole-abdominal irradiation (14 Gy) to model the clinical characteristics of RIC. The mice in the treatment groups received intraperitoneal injections of different doses of CKI for 3 days before exposure to ionising radiation (IR) and were euthanised 7 days after IR. CKI significantly increased the survival rate, body weight, and colon length of IR mice, and it ameliorated the IR-induced tissue damage, intestinal barrier disruption, elevated inflammatory factor levels, macrophage infiltration, and M1 polarisation. Mechanistically, CKI markedly suppressed Piezo1 activation and expression in macrophages as well as the Piezo1-mediated activation of the nuclear factor kappa B (NF-κB) and the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome. Notably, macrophage-specific Piezo1 ablation significantly attenuated the therapeutic efficacy of CKI through impaired mechanosensing, while concomitantly diminishing the magnitude of M1 polarisation and blunting NF-κB/NLRP3 signalling transduction. CONCLUSION:In conclusion, our findings demonstrated that CKI can improve RIC by modulating the Piezo1-mediated NF-κB/NLRP3 pathway and macrophage polarisation, thereby identifying a potential therapeutic target for RIC management and pharmacological intervention.

Lentinus edodes, commonly known as shiitake mushrooms, is widely cultivated for its nutritional and medicinal properties. It has been traditionally used for enhancing immune function, reducing inflammation, and providing antioxidant protection. Among its bioactive components, polysaccharides, particularly β-glucans, have attracted considerable attention for their anti-inflammatory, immunomodulatory, and anticancer effects. However, less is known about α-glucans, which differ in structure and biological activity from β-glucans. This study aims to isolate and characterize an α-glucan from L. edodes (LEP1), investigating its effects on lipopolysaccharide (LPS)-induced systemic inflammation and its potential to modulate gut microbiota in mice. Crude polysaccharides were extracted from L. edodes and purified to obtain LEP1. LEP1 was characterized using various analytical techniques, including scanning electron microscopy (SEM), molecular weight determination, and nuclear magnetic resonance (NMR) analysis. The effects of LEP1 were evaluated in a mouse model of LPS-induced systemic inflammation. Mice were treated with low- and high-doses of LEP1, and various parameters such as body weight, organ indices, and histopathological changes were assessed. The levels of inflammatory cytokines and oxidative stress markers were analyzed, and gut microbiota composition was studied using 16S ribosomal RNA (rRNA) sequencing. LEP1 significantly alleviated LPS-induced weight loss, reduced organ atrophy, and improved colon length. Treatment with LEP1 reduced pro-inflammatory cytokines [interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α] and restored antioxidant enzyme activities [superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)] in liver and colon tissues. Histopathological analysis showed that LEP1 alleviated liver and colon damage caused by LPS, including inflammation and epithelial disruption. LEP1 inhibited the activation of nuclear factor kappa-light-chain enhancer of activated B cells (NF-κB) and NLR family pyrin domain containing 3 (NLRP3) inflammasome pathways while upregulating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) antioxidant pathway. In addition, LEP1 reshaped the gut microbiota by increasing beneficial bacteria, such as Lactobacillus and Allobaculum, while decreasing harmful taxa such as Escherichia-Shigella. LEP1, an α-glucan derived from L. edodes, exhibits significant anti-inflammatory and antioxidant effects in LPS-induced mice, partly through the modulation of key signaling pathways, such as NF-κB/NLRP3 and Nrf2/HO-1. In addition, LEP1 positively affects gut microbiota composition, contributing to its systemic anti-inflammatory effects. These findings indicate that LEP1 shows potential as a functional food ingredient that may mitigate inflammation and oxidative stress, potentially through modulation of immune signaling and microbiota composition. Further studies are required to fully understand the clinical implications of these effects.

Organophosphorus pesticides (OPPs) in foods pose a serious threat to human health, motivating the development of novel analytical methods for their rapid detection and quantification. A magnetic covalent organic framework (M-COF) adsorbent for the magnetic solid-phase extraction (MSPE) of OPPs from foods was reported. M-COF was synthesized by the Schiff base condensation reaction of 1,3,5-tris(4-aminophenyl)benzene and 4,4-biphenyldicarboxaldehyde on the surface of amino-functionalized magnetic nanoparticles. Density functional theory (DFT) calculations showed that adsorption of OPPs onto the surface of M-COF involved hydrophobic effects, van der Waals interactions, π-π interactions, halogen-N bonding, and hydrogen bonding. Combined with gas chromatography-mass spectrometry (GC-MS) technology, the MSPE method features low limits of detection for OPPs (0.002–0.015 μg/L), good reproducibility (1.45%–6.14%), wide linear detection range (0.01–1 μg/L, R ≥ 0.9935), and satisfactory recoveries (87.3%–110.4%). The method was successfully applied for the trace analysis of OPPs in spiked fruit juices.

BACKGROUND:Ulcerative colitis (UC) involves intestinal barrier dysfunction, immune dysregulation, and microbiota imbalance. Bletilla striata polysaccharide (BSP) shows anti-inflammatory potential, but its UC mechanisms remain unclear. PURPOSE:To purify Bletilla striata polysaccharide (BSP-1), characterize its structure, and elucidate its multi-target UC therapeutic mechanisms. METHODS:This study purified a novel structure of neutral glucomannan with a small amount of acetylation (BSP-1). Together with tight junction protein analysis, oxidative stress markers, 16S rRNA sequencing, cytokine quantification, and molecular ecological network examination, the team used a UC mouse model created using dextran sodium sulfate (DSS). RESULTS:BSP-1 alleviated colitis by inhibiting TLR4/MyD88/NF-κB signaling (elevating IL-10; lowering IL-6, TNF-α, and IL-1β) and restoring integrity of the intestinal barrier via upregulating Occludin, ZO-1, MUC2, and Claudin-1. It reduced oxidative stress (lower MDA; higher SOD) and modulated gut microbiota (enriched Lactobacillus, suppressed Escherichia-Shigella). Molecular ecological network analysis showed that BSP-1 restored microbial community stability by enhancing bacterial competition. CONCLUSION:BSP-1 exerts integrated anti-inflammatory, barrier-repairing, and microbiota-regulating effects, highlighting its potential as a multi-target UC therapy and intestinal microecological regulator.

Natural Killer (NK) cells have shown promising prospects in 'off-the-shelf' cell therapy, particularly the NK-92 cell line, which can serve as a foundation for the next generation of universal chimeric antigen receptor (CAR)-engineered NK products. A key strategy for generating universal cellular products is the elimination of the beta-2-microglobulin (B2M) gene, which encodes a component of MHC class I molecules (MHC-I) that plays a role in the presentation of foreign antigens and in the 'licensing' or 'education' of NK cells. To functionally study the impacts of MHC-I deficiency on NK-92, we generated a B2M knockout (KO) NK-92MI (B-92) cell line and compared the multidimensional properties of B2M KO and wild-type NK-92MI cells in terms of biological phenotypes, effector functions, and transcriptomic signatures. We observed a decrease in activating receptors, cytokine production, and cytotoxicity in B-92 cells. Further analysis of signalling events revealed that the upregulated expression and phosphorylation of SHP-1 in B-92 cells inhibited the phosphorylation levels of STAT3 and ERK, thereby affecting their killing function. By knocking out SHP-1 (PTPN6), we partially restored the cytotoxic function of B-92 cells. Notably, we also found that CAR modification can overcome the hyporesponsiveness of B-92 cells. These findings will facilitate further exploration in the development of NK cell-based products.

BACKGROUND:Panax ginseng is a perennial plant valued for its medicinal and nutritional properties. Its fruit contains a variety of bioactive compounds such as ginsenosides, flavonoids, phenolic acids, and anthocyanins. However, the regulatory mechanisms underlying the accumulation of these compounds during fruit development remain largely unexplored. RESULTS:We performed integrated metabolomic and transcriptomic analyses across four developmental stages of ginseng fruit. Metabolite profiling revealed stage-specific accumulation patterns of ginsenosides and phenolics with biphasic trends, and increasing levels of flavonoids and anthocyanins during maturation. We constructed a metabolic and gene expression atlas covering primary metabolism (carbon, amino acids, nitrogen), secondary metabolism (flavonoids, terpenoids), and hormone signaling pathways (abscisic acid, gibberellin, brassinosteroids). Key structural genes and transcription factors, including MYB, bHLH, and ERF families, were found to coordinate stage-specific metabolic shifts. Weighted gene co-expression network analysis revealed metabolite-linked gene modules that delineate regulatory relationships. CONCLUSIONS:This study provides a comprehensive molecular framework of fruit development in P. ginseng, highlighting coordinated transcriptional regulation and metabolic reprogramming. These insights contribute to our understanding of developmental regulation in medicinal plants and lay the groundwork for metabolic engineering strategies aimed at enhancing nutritional quality and bioactive compound production.

This study aimed to develop and validate a nomogram, which can effectively predict the risk of contralateral asymptomatic femoral head collapse in patients with bilateral osteonecrosis of the femoral head (ONFH), undergoing unilateral total hip arthroplasty (THA). We retrospectively analyzed the clinical data of patients who underwent unilateral THA for bilateral non-traumatic ONFH in our center from 2015 to 2018. A total of 103 patients participated in at least 5 years of follow-up. The patients were randomly divided into a training set (70%) and a validation set (30%). Univariate and multivariate Cox analyses were used to determine the independent risk factors for contralateral femoral head collapse. Based on these factors, a predictive nomogram model for 3, 4, and 5 years after THA was developed, and the model was evaluated using receiver operating characteristic (ROC) curve analysis, area under the curve (AUC), decision curve analysis (DCA), and calibration curves. Among the103 patients, 64 patients (62.1%) experienced contralateral femoral head collapse after surgery. Independent risk factors included Japanese investigation committee (JIC) types C1 and C2, lower limb length difference, CE angle, and Harris hip score (HHS) one month after the primary THA. The AUC, calibration curves, and DCA for the predictive model at 3, 4, and 5 years demonstrated good performance of the nomogram. The predictive nomogram model shows good accuracy and clinical utility. Using this tool, clinicians can accurately judge the collapse of the contralateral asymptomatic femoral head after unilateral THA in patients with bilateral ONFH, and they can formulate individualized treatment plans.

As emerging pollutants (EPs) threaten food safety and human health, they demand highly selective and sensitive detection. Because of their excellent permissibility, specific recognition, practicality, and stability, molecularly imprinted polymers (MIPs) are widely used in analyzing EPs in food. We highlight MIP applications across key platforms in sample pretreatment, sensors, chromatographic separation, membrane separation, and simulated enzymes, offering practical references for constructing a method for analyzing food contaminants possessing low detection limits and high recovery rates. This review uniquely integrates MIP design strategies with advanced detection technologies, critically analyzing polymerization methods and imprinting parameters to enable efficient MIP synthesis. It elaborates on recent progress in utilizing MIPs as selective identification units for analyzing EPs in food, categorized according to different detection technologies. Finally, the review concludes with challenges and future trends to advance MIP-based early warning systems for food safety.

ETHNOPHARMACOLOGICAL RELEVANCE:Kaji-ichigoside F1 (KF1), the main active component of the Guizhou ethnic medicinal material Rosa roxburghii Tratt, is widely used in China due to its anti-inflammatory properties. However, the protective effects of KF1 against drug-induced liver injury and its potential mechanisms are not yet understood. AIM OF THE STUDY:We aimed to investigate the effects of KF1 on acute liver injury (ALI) and explore its underlying mechanisms, particularly its role in modulating the gut microbiota to inhibit ALI development. MATERIALS AND METHODS:KF1 was prepared via 80 % ethanol extraction, silica gel column chromatography, and Sephadex LH-20 column chromatography. Mouse models of ALI were established using acetaminophen (APAP) treatment, with or without KF1 (5 and 10 mg/kg). 16S rRNA gene sequencing, metabolomics, and transcriptomics approaches were employed to explore the inhibitory effect of KF1 on ALI. Additionally, the role of the gut microbiota was investigated through antibiotic treatment and fecal microbiota transplantation experiments. RESULTS:Treatment with KF1 significantly altered the gut microbiota composition, notably increasing the abundance of the probiotic Akkermansia muciniphila (A. muciniphila). Furthermore, A. muciniphila enhanced the levels of beneficial metabolites, including inosine. Notably, inosine significantly suppressed inflammatory factors and improved APAP-induced ALI. Transcriptomic analysis revealed that inosine inhibited key signaling pathways, including MAPK, PI3K-AKT, JAK-STAT3, IL-17, TNF, and cytokine-cytokine receptor interactions. Importantly, the preventive effect of KF1 is dependent on microbial mechanisms. CONCLUSION:KF1 protects against ALI by modulating the gut microbiota and associated metabolites, thereby promoting a more favorable state and inhibiting pro-inflammatory pathways.

Type 2 diabetes mellitus (T2DM) is a global health challenge with limited efficacy of current treatments, necessitating alternative therapies. Plant-derived pectin, composed of galacturonic acid and structural domains such as homogalacturonan, has shown promise as an anti-diabetic agent. Pectin exerts its therapeutic effects through multiple mechanisms, including enhancing β-cell function, regulating glucose metabolism, improving insulin sensitivity, inhibiting digestive enzymes, and restoring gut microbiota balance. Its bioactivity is influenced by physicochemical properties like molecular weight, degree of methylation, and structural complexity. This review explores the anti-diabetic potential of pectin, its structure-activity relationships, and mechanisms of action, providing insights for its development as a novel therapeutic agent in T2DM management.

The treatment of gout remains a huge challenge, and currently, the mainstream Western medicine treatment methods still have some drawbacks. This study aims to elucidate the potential mechanism of modified Simiao decoction (MSMD) in the treatment of gout through network pharmacology analysis, Mendelian randomization (MR), and molecular docking. We hope to contribute to the research on the targets of action of traditional Chinese medicine and the exploration of the mechanism of gout. We employed network pharmacology to screen the potential targets of MSMD in relation to gout. MR was utilized to further augment the potential targets and elucidate the causal associations between them and gout. Eventually, the identified targets were combined with active ingredients for the purpose of molecular docking. Through network pharmacology, we identified 44 potential targets. MR analysis identified 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) as a potential causal risk factor for gout (odds ratio [95% confidence interval] = 1.192 [1.033-1.375], P = .016). Molecular docking predicted that TP53, IL6, TNF, peroxisome proliferator-activated receptor γ, IL1B, and HMGCR exhibited favorable binding effects with the active ingredients. TP53, IL6, TNF, peroxisome proliferator-activated receptor γ, and IL1B are central inflammatory targets of MSMD. Notably, HMGCR was identified as a causal risk factor for gout and a potential target of MSMD. We hypothesize that MSMD may exert therapeutic effects by inhibiting HMGCR activity, counteracting its risk effect. This hypothesis, along with the anti-inflammatory roles of other targets, warrants further experimental validation.

Non-small cell lung cancer (NSCLC) with PIK3CA mutations demonstrates significant challenges in treatment due to enhanced bone metastasis and immune checkpoint resistance. This study investigates the efficacy of tumor-targeting peptide 1-modified cancer stem cell-derived extracellular vesicles (TMTP1-TSRP-EVs) in reshaping the tumor microenvironment and reversing immune checkpoint resistance in NSCLC. By integrating TMTP1-TSRP into EVs, we aim to specifically deliver therapeutic agents to NSCLC cells, focusing on inhibiting the PI3K/Akt/mTOR pathway, a crucial driver of oncogenic activity and immune evasion in PIK3CA-mutated cells. Our comprehensive in vitro and in vivo analyses show that TMTP1-TSRP-EVs significantly inhibit tumor growth, reduce PD-L1 expression, and enhance CD8+ T cell infiltration, effectively reversing the immune-suppressive microenvironment. Moreover, the in vivo models confirm that our approach not only suppresses bone metastases but also overcomes primary resistance to immune checkpoint inhibitors by modulating the expression of key immunological markers. These findings suggest that targeted delivery of TMTP1-TSRP-EVs could provide a novel therapeutic strategy for treating PIK3CA-mutant NSCLC, offering significant improvements over traditional therapies by directly targeting the molecular pathogenesis of tumor resistance and metastasis.