Epoxy resins (EP) as structural polymeric materials (e.g. adhesives, coatings) have been broadly utilized in the fields of civil construction, electrical insulation, aerospace, etc. However, EP suffers from the high flammability, low fire safety, and degraded mechanical properties after flame-retardant modification. Herein, epoxy-based materials integrated with excellent flame-retardant efficacy, high fire safety simultaneously with early-warning functionality enabled by thermochromism, and strong mechanical properties have been proposed. A fluorescent flame-retardant hybrid (APP@ZnQ2) was rationally designed through the assembly of quinolone-zinc complex (ZnQ2) on the ammonium polyphosphate (APP) surface, and then fed into epoxy-curing system for the construction of multifunctional epoxy-based composites. The resulting composites exhibited a significant thermochromic behavior, enabled by a distinct change in their characteristic chartreuse fluorescence when temperatures exceeded 160 degrees C, thus realizing the early-warning functionality. The epoxy composite containing 6 wt % APP@ZnQ2 was imparted with an LOI value up to 29.6% and the highest V-0 rating. Compared to pure EP, its heat/smoke parameters including the PHRR defined as Peak Heat Release Rate, THR (Total Heat Release), and TSP (Total Smoke Production) values were remarkably decreased by 72%, 67%, and 72%, respectively. Beyond that, because of the improved compatibility of APP@ZnQ2, the composite possessed comparable mechanical strength, especially in impact strength, which surpassed that of pure EP. Generally, this work develops a straightforward yet effective strategy to fabricate epoxy composites with both high fire safety and early-warning functionality.

BackgroundChimeric antigen receptor (CAR) T-cell therapy has shown limited efficacy against solid tumors due to antigen heterogeneity and scarcity of tumor-specific targets.MethodsTo address those challenges, we report a covalent CAR-T strategy that achieves programmable tumor recognition via oncolytic adenovirus-mediated (OAD) delivery of artificial antigens. An engineered OAD was designed to induce tumor-selective expression of a membrane-anchored SpyTag-containing artificial antigen on infected tumor cells. In parallel, we generated SpyCatcher CAR-T cells by replacing the conventional single-chain variable fragment (scFv) with SpyCatcher, which forms a spontaneous covalent bond with SpyTag and redirects CAR-T-cell activity toward virus-labeled tumor cells.ResultsIn vitro, optimized SpyCatcher CAR-T cells mediated selective cytotoxicity against SpyTag-positive tumor cells, achieving >85% specific lysis at an effector-to-target ratio of 1:1, while sparing antigen-negative cells. The OAD efficiently induced tumor-selective expression of membrane-anchored SpyTag-fused antigens across multiple cell lines. Combined treatment with OAD and SpyCatcher CAR-T cells resulted in substantially greater antitumor activity than either monotherapy alone. In vivo, the combinatorial strategy significantly inhibited tumor growth and increased intratumor CD3+, CD8+ T-cell infiltration in both immunodeficient and immunocompetent mouse models. Importantly, patient-derived prostate cancer organoids were effectively transduced by OAD and supported robust SpyCatcher CAR-T cell infiltration and cytotoxicity, demonstrating the translational potential of this approach.ConclusionsThis study establishes a modular platform for solid tumor immunotherapy therapy by integrating covalent SpyCatcher CAR-T cells with SpyTag-delivering OAD. By decoupling tumor recognition from endogenous antigen expression, this approach provides a generalizable strategy to overcome antigen heterogeneity and scarcity of tumor-specific targets in solid tumors.

We developed an HSC-targeted nanoplatform using a mitochondrial H 2 O 2 /viscosity probe and melatonin. It enables real-time imaging of hepatic ischemia-reperfusion injury and ROS scavenging to mitigate damage, offering a cell-specific theranostic tool.

Chimeric antigen receptor (CAR) T-cell therapy is a new type of highly precise and targeted immunotherapy for urological diseases. It has demonstrated significant therapeutic potential in chronic and autoimmune kidney diseases such as renal fibrosis and membranous nephropathy. However, owing to its high cost, low efficiency, and serious side effects, such as cytokine storm syndrome, a new generation of drugs has emerged to solve these problems. In vivo CAR T-cell therapy is a treatment method that directly modifies specific cells within the patient’s body through various delivery systems. Nonviral vectors (nanoparticles and exosomes) and viral vectors (adeno-associated viruses and lentiviruses) can be engineered to achieve better therapeutic effects. By taking advantage of different delivery systems and minimizing their drawbacks, in vivo CAR T-cell therapy can improve the stability and targeting ability, reduce immunogenicity, and minimize side effects. This review summarizes the mechanisms of action and clinical applications of various delivery systems used in in vivo CAR T-cell therapy, highlighting their potential in the treatment of urological diseases. Through a deeper understanding of the construction and optimization of well-designed platforms, the development of optimal delivery systems has valuable implications for the establishment of new pharmaceuticals for in vivo CAR T-cell therapy in urological diseases.

Background: Chimeric antigen receptor (CAR) T-cell therapy is clinically limited by lentiviral vector dependence, ex vivo activation/expansion requirements, complex processes, high costs, and impaired antitumor functions. For advanced cancer, the long manufacturing cycle of traditional CAR-T cells further compromises the treatment timeliness, creating an urgent need for optimized preparation strategies. Materials and methods: A gene delivery system was constructed using microtubule-associated sequence and nuclear localization signal peptide–modified poly(β-amino ester) nanoparticles, combined with microfluidic technology. Nonactivated primary human T cells were transfected without ex vivo activation. The transfection efficiency, gene expression persistence, and in vitro cytotoxicity against the B7-H3-positive bladder cancer cell line T24 were evaluated. Results: The system achieved gene delivery to nonactivated T cells within 2 to 6 hours, with a transfection efficiency that was 7-fold higher than that of unmodified nanoparticles. Target genes were stably expressed for over 5 days. The resulting CAR-T cells exhibited potent, specific cytotoxicity against T24 cells, avoiding viral vector risks and reducing manufacturing costs. Conclusions: This strategy innovates CAR-T-cell preparation by targeting nonactivated T cells and overcoming traditional bottlenecks. This provides a simpler, faster, cost-effective, and safe approach for adoptive cellular immunotherapy with significant translational value for bladder cancer and other solid tumors.

Gene therapy offers significant potential for precision medicine, yet its clinical translation is often hindered by the lack of delivery systems that simultaneously achieve high efficiency, stability, and safety. Poly(β-amino esters) (PBAEs), as promising non-viral vectors, face challenges such as insufficient serum stability and limited transfection efficacy, particularly in hard-to-transfect primary immune cells. To address these limitations, this study developed a microfluidic co-assembly strategy to incorporate polyethylene glycol (PEG) or polyethylene glycol-anchored lipid (PEG-lipid) into PBAE nanoparticles. Compared to the PBAE-only controls, the optimized PEG/PEG-lipid-incorporated nanoparticles exhibited three key advantages: (1) maintained superior transfection efficiency across diverse cell types, even under stringent conditions with reduced DNA doses or lower PBAE/DNA ratios; (2) significantly enhanced serum stability; and (3) successfully overcame the transfection barrier in non-activated primary T cells, with further efficiency gains achieved through conjugation with targeting peptides and antibodies. The integrated microfluidic assembly technology and material functionalization strategy presented in this study provide robust technical support for the development of efficient and low-toxicity non-viral gene vectors. This integrated strategy offers a novel approach for advancing the clinical translation of immune cell gene therapies.

Liver fibrosis is a pathological repair response to chronic liver injury and may progress to cirrhosis, liver failure, or hepatocellular carcinoma if untreated. Currently, no approved therapies specifically target advanced liver fibrosis, thus exploring the molecular mechanisms underlying liver fibrosis has become crucial. Previous studies have highlighted significant controversy regarding the role of ferroptosis in liver fibrosis. Given that reactive oxygen species (ROS) serve as key mediators to both processes, ROS may serve as a molecular nexus connecting ferroptosis and liver fibrosis. To comprehensively elucidate the molecular network involving ROS and ferroptosis in liver fibrosis, we designed and synthesized the first multi‐functional “cocktail” fluorescence probe, FP‐ROS, enabling highly sensitive and selective simultaneous imaging of O 2 •− , H 2 O 2 , and ONOO − . FP‐ROS was successfully employed to assess ferroptosis levels in the livers of fibrosis mice following drug intervention. Combining transcriptomic and proteomic analyses, we elucidated the signaling pathway NOX→ONOO − →GCLM(C46)→GSH→ferroptosis→hepatic stellate cells (HSCs) activation. This study demonstrates that ferroptosis plays a critical role in HSCs activation and further elucidates the molecular interplay between ROS and ferroptosis in fibrosis progression. These findings provide novel insights into the diagnosis and therapeutic strategies for liver fibrosis.

Tumor organoids mimicking the tumor microenvironment (TME) are key tools for tumor immunity research and personalized cancer therapy development. We integrated microgravity culture with microfluidic chip technology (Micro-GRA& FLU) to establish a platform for evaluating chimeric antigen receptor (CAR)-γδ T cell efficacy under physiological-like conditions. Patient-derived glioblastoma (GBM) cells were microgravity-cultured into glioblastoma organoids (GBOs). Pathological analysis validated GBO similarity to matched GBM in immune cell phenotypes. Microfluidic chips assessed CAR-γδ T cell cytotoxicity against GBOs. The low-cost, easy-to-operate microgravity system generated viable, uniform GBOs that retained GBM TME features. CAR-γδ T cells showed strong cytotoxicity against GBOs in microfluidic chips; individualized combination therapy enhanced their antitumor activity vs. monotherapy. This study establishes a scalable, physiologically relevant Micro-GRA& FLU platform for evaluating CAR-γδ T cell therapies in GBM organoids. To evaluate the efficacy of CAR-γδ T cells for GBM, this study developed the Micro-GRA&FLU platform. This low-cost, scalable platform provides a physiologically relevant model for assessing personalized immunotherapy for GBM.

D-2-Hydroxyglutarate (D-2-HG) is a functional endogenous metabolite in various domains of life. Its abnormal accumulation promotes human tumorigenesis. Convenient D-2-HG testing for diagnosis and prognosis of D-2-HG-related diseases remains technically challenging, and there is no analytical method to directly detect D-2-HG in living cells. Here, we identify a D-2-HG-specific transcriptional activator, HgcR, and develop a d-2-HG sensor (DHOR) using HgcR as a sensing moiety. Then, we build a portable device adaptive with DHOR for rapid and low-cost point-of-care D-2-HG testing in serum, urine, and glioma tissue samples. DHOR also allow spatiotemporal resolution of D-2-HG in living bacteria and human cells. We use DHOR to identify D-2-HG transporters from Escherichia coli and human solute carrier 22 family. Overall, DHOR provides a powerful and versatile tool for in vitro and live-cell detection of D-2-HG, offering the opportunity to deepen our understanding about physiological and pathogenetic roles of D-2-HG.

Urological tumors represent a significant global health challenge, with conventional therapies often proving insufficient to control disease progression. Recent breakthroughs in cellular immunotherapy, particularly in chimeric antigen receptor (CAR)-T cell, CAR-natural killer cell, and CAR-macrophage therapies, have demonstrated remarkable potential for treating these malignancies. Ongoing research is actively refining CAR-based strategies to enhance their precision in targeting tumor-associated antigens. This review comprehensively summarizes the applications of CAR cell therapy in the following 3 major urological tumors: renal cell carcinoma, bladder cancer, and prostate cancer. Furthermore, we analyzed the current advantages and limitations of these approaches and propose potential strategies for optimization focused on CAR-T cells. This review will provide future directions in this field and contribute to the development of more effective treatments for patients with urological cancer.

Various challenges, including tumor heterogeneity and inadequate T cell infiltration, impede the progress of chimeric antigen receptor T cell (CAR-T) therapy for glioblastoma (GBM). To address these obstacles, a multiple step strategy is designed. Initially, literature review and bioinformatics analysis to screen a set of antigens that are heterogeneously expressed in GBM, which are designated as the target-bank, are leveraged. Then, according to the multiplex immunohistochemistry results of each patient's tumor sample, a personalized panel of antigens based on the principle that most cancer cells in tumor tissues can be covered from the target-bank is selected. To target these antigens, Vδ1 T cells are chosen as CAR vehicles because of its high tissue infiltration and off-the-shelf properties, and an optimized protocol for engineering CAR-Vδ1 T cells with high purity and cytotoxicity, low exhaustion, and cytokine release is developed. Next, the specific panel of cocktail CAR-Vδ1 T cells in the GBM organoids that are directly derived from the same patient's tumor is tested. The term "prof" cocktail therapy is coined to describe the approach using precise and rational combination of tumor antigens, organoid-based evaluation, and fitness of Vδ1 T cells. It may accelerate development of effective CAR-T drugs for heterogeneous solid tumors.

Hepatic ischemia-reperfusion injury (HIRI) is the main cause of postoperative liver dysfunction and liver failure. Traditional separation of HIRI diagnosis and therapy confers several disadvantages, including the inability to visualize the therapeutic and asynchronous action. However, developing a versatile material with integrated diagnosis and treatment for HIRI remains a great challenge. Given that hypochlorous acid (HOCl) plays a crucial oxidative role in HIRI, we developed a single-component multifunctional fluorescent theranostic platform (MB-Gly) with a "three-in-one" molecular design incorporating a near-infrared fluorophore methylene blue, glycine and a HOCl-response unit, which could not only provide real-time visualization of HIRI but also boost targeted drug delivery. Using MB-Gly, we were able to achieve real-time and dynamic monitoring of HOCl during HIRI in hepatocytes and mouse livers and reduce the liver damage in hepatocytes and mice. RNA sequencing illustrated the therapeutic role of MB-Gly associated with changes in gene expression related to apoptosis, oxidative stress, metabolism and inflammation. To the best of our knowledge, this is the first multifunctional fluorescent theranostic system for HIRI reported to date. Our smart "three-in-one" approach shines light on the etiology and pathogenesis of HIRI, providing profound insights into the development of potential therapeutic targets.

Abstract Background Glomerular endothelial cell (GENC) injury would be a characteristic of early stage diabetic nephropathy (DN) and the investigation of potential therapeutic targets for preventing GENC injury has clinical importance. Methods DN was induced in C57BL/6J mice by intraperitoneal injection of streptozotocin. GENC was transfected with plasmid containing siRNA-β-arrestin-2, shRNA-ATF6, pCDNA-β-arrestin-2 or pCDNA-ATF6. Additionally, we administrated adeno-associated virus (AAV) containing shRNA-β-arrestin-2 via tail vein injection in DN mice. Results The upregulation of β-arrestin-2 was observed in DN patients as well as in GENC from DN mice. Knockdown of β-arrestin-2 reduced endoplasmic reticulum stress (ER stress) and apoptosis in high glucose treated GENC which were reversed by overexpression of activating transcription factor 6 (ATF6). Moreover, overexpression of β-arrestin-2 led to the activation of ER stress and the apoptosis of GENC which could be mitigated by silencing of ATF6. Furthermore, knockdown of β-arrestin-2 by the administration of AAV-shRNA-β-arrestin-2 had alleviated renal injury in DN mice. Conclusions This study offer novel perspectives on the crucial involvement of β-arrestin-2 in GENC injury. Knockdown of β-arrestin-2 prevents GENC apoptosis by inhibiting ATF6-mediated ER stress in vivo and vitro. Consequently, β-arrestin-2 may represent a promising therapeutic target for the clinical management of patients with DN.

Current cytotoxic T lymphocyte (CTL) activating immunotherapy requires a major histocompatibility complex I (MHC-I)-mediated presentation of tumor-associated antigens, which malfunctions in around half of patients with triple-negative breast cancer (TNBC). Here, we create a LCL161-loaded macrophage membrane decorated nanoparticle (LMN) for immunotherapy of MHC-I-deficient TNBC. SIRPα on the macrophage membrane helps LMNs recognize CD47-expressing cancer cells for targeted delivery of LCL161, which induces the release of high mobility group protein 1 and proinflammatory cytokines from cancer cells. The released cytokines and high mobility group protein 1 activate antitumor immunity by increasing the intratumoral density of the phagocytic macrophage subtype by 15 times and elevating the intratumoral concentration of CTL lymphotoxin by 4.6 folds. LMNs also block CD47-mediated phagocytosis suppression. LMNs inhibit the growth of MHC-I-deficient TNBC tumors, as well as those resistant to combined therapy of anti-PDL1 antibody and albumin-bound paclitaxel, and prolong the survival of animals, during which process CTLs also play important roles. This macrophage membrane-decorated nanoparticle presents a generalizable platform for increasing macrophage-mediated antitumor immunity for effective immunotherapy of MHC-I-deficient cancers.

Early diagnosis and precise treatment of hepatocellular carcinoma (HCC) are crucial for human health. Therefore, addressing the potential markers of HCC, glutathione (GSH) and viscosity, we constructed a fluorescent probe (PG-V) activated cascadically by GSH/viscosity. PG-V possessed excellent photophysical properties and biocompatibility, and could specifically illuminate tumor tissue, achieving fluorescence imaging of HCC, and imaging-guided tumor resection.

Isocitrate dehydrogenase mutation-induced D-2-hydroxyglutarate (D-2-HG) accumulation may promote the tumorigenesis of numerous cancers. D-2-HG produced by several promiscuous enzymatic reactions under normal conditions is also toxic to organisms. D-2-HG was thus previously viewed as an abnormal metabolite with no physiological function and D-2-HG conversion to 2-ketoglutarate by D-2-HG dehydrogenase seems to be a process of metabolite repair. However, several recent studies have revealed that D-2-HG production is not a meaningless process without a physiological purpose. D-2-HG production actually participates in many core metabolic processes, and disorders of these processes may be the reason for the D-2-HG accumulation in some tumors lacking the isocitrate dehydrogenase mutation. This review summarizes the diverse and widely distributed metabolic processes involving D-2-HG, highlights the physiological functions of D-2-HG generation from various precursors, and discusses the possible applications of enzymes related to D-2-HG metabolism in synthetic biology, diagnosis, and treatment of D-2-HG accumulation-induced diseases.

L-2-Hydroxyglutarate (L-2-HG) is a functionally compartmentalized metabolite involved in various physiological processes. However, its subcellular distribution and mitochondrial transport remain unclear owing to technical limitations. In the present study, an ultrasensitive L-2-HG biosensor, sfLHGFR(H), composed of circularly permuted yellow fluorescent protein and L-2-HG-specific transcriptional regulator, is developed. The ability of sfLHGFR(H) to be used for analyzing L-2-HG metabolism is first determined in human embryonic kidney cells (HEK293FT) and macrophages. Then, the subcellular distribution of L-2-HG in HEK293FT cells and the lower abundance of mitochondrial L-2-HG are identified by the sfLHGFR(H)-supported spatiotemporal L-2-HG monitoring. Finally, the role of the l-glutamate transporter SLC1A1 in mitochondrial L-2-HG uptake is elucidated using sfLHGFR(H). Based on the design of sfLHGFR(H), another highly sensitive biosensor with a low limit of detection, sfLHGFR(L), is developed for the point-of-care diagnosis of L-2-HG-related diseases. The accumulation of L-2-HG in the urine of patients with kidney cancer is determined using the sfLHGFR(L) biosensor.

D-2-Hydroxyglutarate (D-2-HG) is an oncometabolite aberrantly accumulated in patients with D-2-hydrox-yglutaric aciduria or various isocitrate dehydrogenases mutation-associated cancers. The detection of D-2-HG is of great significance for the diagnostic and therapeutic aspects of these diseases. Herein, a Fo center dot rster resonance energy transfer (FRET)-based D-2-HG biosensor was developed. The D-2-HG-specific allosteric transcription factor DhdR from Achromobacter denitrificans NBRC 15125 was used as its biorecognition element and inserted between two fluorescent proteins Clover and mRuby2. The response magnitude of the biosensor was improved by trun-cation of the terminal amino acids of DhdR and addition of artificial linkers. The optimized variant DHGFR1.0 exhibited excellent sensitivity and specificity with a maximum emission ratio change (Delta Rmax) of 30.77%, a half -maximal effective concentration (EC50) of 2.78 mu M, and a dynamic detection range of 0.17-36.22 mu M. It was able to detect the concentration of D-2-HG in body fluids and cell-related samples with high accuracy and precision in vitro. The ligand-binding site of DhdR was predicted and several biosensors with low affinity for D-2-HG were also generated by site-directed mutagenesis of DhdR.

Bio-based hydrophobic coating modified cotton fabrics with durable flame retardancy are of high interest in the application of oil-water separation for not only avoiding the use of hazardous substances but also improving the fire safety during use. Herein, phytic acid@Polyurushiol‑titanium complex coated cotton fabric was developed using the facile dip-coating method involving the sequential immersion in the solution of poly(ethyleneimine), phytic acid, titanium oxide, and urushiol. The underlying coating accommodated abundance of phytic acid, which imparted excellent flame retardancy to cotton fabric, and the top coating composed of the polyurushiol‑titanium complex endowed cotton fabric with high hydrophobicity that the water contact angle (WCA) was up to 149.8°. The hydrophobicity also guaranteed effective protection of the underlying phytic acid against chemical solvents and abrasion. Besides, the hydrophobic coating allowed cotton fabric for good self-cleaning and effective oil-water separation. Therefore, the preparation of phytic acid@polyurushiol‑titanium complex coated cotton fabric offers a promising approach to construct durable biomass-coated cellulose-based fabric with multifunctionality.

The oncometabolite D-2-hydroxyglutarate (D-2-HG) has emerged as a valuable biomarker in tumors with isocitrate dehydrogenase (IDH) mutations. Efficient detection methods are required and rapid intraoperative determination of D-2-HG remains a huge challenge. Herein, D-2-HG dehydrogenase from Achromobacter xylosoxidans (AX-D2HGDH) was found to have high substrate specificity. AX-D2HGDH dehydrogenizes D-2-HG and reduces flavin adenine dinucleotide (FAD) bound to the enzyme. Interestingly, the dye resazurin can be taken as another substrate to restore FAD. AX-D2HGDH thus catalyzes a bisubstrate and biproduct reaction: the dehydrogenation of D-2-HG to 2-ketoglutarate and simultaneous reduction of non-fluorescent resazurin to highly fluorescent resorufin. According to steady-state analysis, a ping-pong bi-bi mechanism has been concluded. The Km values for resazurin and D-2-HG were determined as 0.56 mu M and 10.93 mu M, respectively, suggesting high affinity to both substrates. On the basis, taking AX-D2HGDH and resazurin as recognition and fluorescence transducing element, a D-2-HG biosensor (HGAXR) has been constructed. HGAXR exhibits high sensitivity, accuracy and specificity for D-2-HG in different biological samples. With the aid of HGAXR and the matched low-cost palm-size detecting device, D-2-HG levels in frozen sections of resected brain tumor tissues can be measured in a direct, simple and accurate manner with a fast detection (1-3 min). As the technique of frozen section is familiar to surgeons and pathologists, HGAXR and the portable device can be easily integrated into the current workflow, having potential to provide rapid intraoperative pathology for IDH mutation status and guide decision-making during surgery.