ETHNOPHARMACOLOGICAL RELEVANCE:Mulberry (Morus alba L.) has a longstanding history of use in Traditional Chinese Medicine (TCM). It is recorded in the classic Compendium of Materia Medica (Ben Cao Gang Mu) that mulberry possesses the efficacy to "strengthen muscles and bones". This traditional indication suggests a potential role in skeletal health, yet its scientific and pharmacological basis has not been fully elucidated. The present study provides the first experimental evidence that a standardized polyphenol extract from mulberry (ABRU) bidirectionally regulates bone metabolism, simultaneously promoting bone formation and inhibiting bone loss. This work not only validates a centuries-old ethnopharmacological claim with modern molecular and functional evidence but also identifies mulberry polyphenols as a promising candidate for developing natural product-based therapies for bone metabolic disorders like osteoporosis. AIM OF THE STUDY:This study aims to investigate the dual regulatory effects and mechanisms of mulberry polyphenols (ABRU) on bone metabolism. METHODS:MC3T3 cells and Bone marrow-derived macrophages (BMDMs) were used to evaluate the effects of various mulberry extracts on osteoblastic mineralization and osteoclastogenesis. Active extracts were analyzed for their chemical composition using liquid chromatography-mass spectrometry (LC-MS). Network pharmacology, molecular docking, ROC analysis, and thermal shift assays were employed to identify the molecular targets of ABRU. The dual regulation of bone metabolism by ABRU was further assessed in vivo using subcutaneous bone formation assays and a D-galactose-induced mouse model of bone loss. RESULTS:In vitro experiments demonstrated that mulberry polyphenol (ABRU) significantly enhanced osteoblast proliferation and differentiation. After 24 h, osteoblast proliferation doubled, after 14 days, mineralization (measured by Alizarin Red staining) and alkaline phosphatase activity both increased two-fold. ABRU also inhibited osteoclast function, achieving 50 % suppression within 6 days, indicating a dual regulatory effect on bone metabolism-promoting bone formation while inhibiting resorption. Further investigation using Pparg knockdown revealed that silencing Pparg reduced SOST expression and upregulated osteogenic markers ALP, OCN, RUNX2. Crucially, Pparg suppression activated the Wnt signaling pathway, evidenced by increased LRP5/TCF expression and enhanced nuclear accumulation of β-catenin.In vivo, oral administration of ABRU dose-dependently increased subcutaneous bone formation in nude mice by 2-4 fold. In the D-galactose-induced aging bone loss model, ABRU supplementation improved bone mineral density by ∼20 %, increased trabecular bone area by 24.5 %, and reduced osteoclast number by 58.4 %. Mechanistically, the core polyphenolic components of ABRU inhibit PPARG expression, leading to downregulation of sclerostin (SOST) activity. This molecular pathway supports the dual regulation of bone metabolism by ABRU. CONCLUSION:This study is the first, through both in vitro and in vivo experiments, to demonstrate that mulberry polyphenols (ABRU) promote bone formation and alleviate age-related bone loss. It also reveals the dual regulatory effects of ABRU on osteoblast differentiation and osteoclast function, mediated via the PPARG/SOST signaling axis.

Fracture healing is a complex process driven by endogenous regenerative mechanisms, with early biological responses playing a pivotal role in determining healing outcomes. During this critical phase, the body establishes a dynamic equilibrium across multiple systems, akin to the precise calibration of a biological clock. The inflammatory response is tightly regulated through the interplay of pro- and anti-inflammatory signals, ensuring efficient immune cell recruitment for necrotic tissue clearance while preventing excessive inflammation that could compromise surrounding tissues. Simultaneously, the coagulation cascade maintains a delicate balance between clot formation and anticoagulation, facilitating hemostasis and repair initiation while mitigating thrombotic risks. Energy metabolism is similarly fine-tuned, with coordinated anabolic and catabolic activity providing the necessary substrates and energy for regeneration. These interconnected processes collectively drive the phenotypic transformation of cells from diverse lineages, ultimately shaping the trajectory of fracture healing. In this review article, we propose an integrated ‘biological orchestration’ framework. Rather than viewing these systems in isolation, we discuss the intricate crosstalk among inflammatory homeostasis, coagulation balance, and metabolic adaptation. Additionally, we provide a multi-dimensional exploration of the fracture healing process, encompassing the microenvironment, intra-osseous dynamics, and the regulatory influence of surrounding tissues. By elucidating the temporal orchestration of these systems, this review offers theoretical insights that may inform the development of precise therapeutic strategies for bone regeneration.

OBJECTIVE:Brachytherapy (BT) is favored over external beam radiation therapy (EBRT) for certain tumors with modest ionizing damage. However, the biological effect of BT in glioblastoma remains uncertain. Thus, this study aimed to compare BT and EBRT in glioblastoma treatment. METHODS:BT cell culture templates ensured equal in vitro doses between BT and EBRT. The effects of EBRT and BT on GL261 glioblastoma cells were detected by CCK8 assay, colony formation assay, and flow cytometry. In addition, cellular reactive oxygen species (ROS) levels and mitochondrial membrane potential were measured. Western blotting was used to verify the mechanisms underlying regulated cell death, including apoptosis and ferroptosis. A GL261 xenograft model was also constructed for in vivo validation. RESULTS:BT significantly reduced GL261 viability and proliferation while promoting apoptosis. It also significantly increased ROS levels and altered MMP. Moreover, BT upregulated the level of γ-H2AX (DNA damage), GPX4, SLC7A11 (antioxidant defense), and PTGS2 (ferroptosis marker), indicating BT-induced ferroptosis and an adaptive cellular antioxidant response. In xenografts, BT significantly inhibited tumor growth, decreased CD31 expression levels indicating impaired angiogenesis, and increased HIF-1α levels reflecting exacerbated tumor hypoxia. CONCLUSION:BT effectively induces oxidative stress, DNA damage, and ferroptosis in glioblastoma. It also triggers a robust antioxidant defense response while influencing angiogenesis and the hypoxic tumor microenvironment.

Proteolysis-targeting chimeras (PROTACs) represent a transformative therapeutic modality that leverages the endogenous ubiquitin-proteasome system (UPS) to achieve targeted protein degradation. These heterobifunctional molecules facilitate the recruitment of E3 ubiquitin ligases to a protein of interest (POI), promoting its ubiquitination and subsequent proteasomal degradation. In contrast to conventional inhibitory approaches, PROTACs operate catalytically, enabling the degradation of a wide spectrum of targets—including those harboring drug-resistant mutations—at significantly lower doses. These attributes have positioned PROTACs as promising agents, particularly in oncology, where their efficiency and broad applicability have been increasingly demonstrated. Nonetheless, clinical translation of PROTACs faces challenges such as poor bioavailability, insufficient tumor-specific accumulation, and off-target effects. The integration of nanomedicine-based delivery platforms offers a viable path to overcome these limitations by enhancing drug stability, improving tissue selectivity, and reducing systemic toxicity. This review outlines the rational design principles underlying nano-PROTACs, highlights recent advances in nanoformulations aimed at optimizing their delivery and efficacy, discusses emerging combination regimens and innovative design strategies, and critically assesses the translational challenges and future directions of nano-PROTACs. Overall, nano-PROTAC technology has pioneered a brand-new approach in the field of disease treatment, providing unprecedented opportunities for precision medicine and personalized disease treatment.

ABSTRACT The tumor microenvironment (TME) represents a highly specialized niche that not only supports malignant progression but also serves as a rich source of therapeutic targets. Characterized by distinctive pathophysiological features, including aberrant enzymatic activity, reductive stress, oxygen deprivation, and oxidative stress, the TME creates a unique biochemical landscape that can be exploited for targeted therapeutic interventions. In recent years, proteolysis‐targeting chimeras (PROTACs) have emerged as a revolutionary therapeutic paradigm in precision oncology, offering unprecedented capabilities for targeted protein degradation through hijacking the ubiquitin‐proteasome system. However, the clinical translation of conventional PROTACs faces significant challenges, particularly their inability to distinguish between malignant and healthy tissues, leading to potential off‐target effects and dose‐limiting toxicities. To address these limitations, a new generation of smart PROTAC prodrugs has been developed, designed to remain pharmacologically inert until selectively activated by TME‐specific stimuli. These innovative prodrug strategies employ various triggering mechanisms, including: (1) enzyme‐cleavable masking groups responsive to tumor‐associated proteases/esterases; (2) GSH‐sensitive disulfide linkages that exploit the reductive intracellular environment; (3) hypoxia‐activated prodrug moieties targeting oxygen‐deprived tumor regions; (4) ROS‐labile functional groups that respond to oxidative stress; and (5) pH‐responsive groups that react to the acidic TME. This comprehensive review systematically examines recent advances in TME‐responsive PROTAC prodrug design, with particular emphasis on structure‐activity relationships, activation kinetics, and therapeutic efficacy. Furthermore, we discuss emerging combination strategies that synergize TME‐responsive PROTACs with other treatment modalities, as well as current challenges and future directions in this rapidly evolving field.

A single-material system that seamlessly combines bioaerosol sampling, subsequent pathogen detection, and on-demand inactivation represents a pivotal but unmet goal for intelligent environmental health monitoring. Herein, a Janus air filter membrane (AIE-LGFM) was developed with fully biobased lignin-gelatin for this specific purpose. This membrane is constructed through a green phase-separation process, forming a water-dissolvable and tunable porous network with a high interception efficiency (99.4%) and low pressure drop (40 mbar). Significantly, a gemini-type aggregation-induced emission (AIE) surfactant (G-MeOTTVP) was synthesized, featuring a symmetric cationic structure that confers superior surface activity and bacterial targeting ability, alongside strong near-infrared fluorescence emission (755 nm) and a high reactive oxygen species yield. This AIE gemini surfactant is asymmetrically coated onto the membrane, creating a Janus architecture that spatially separates its functions. The functionalized front side serves as a strong and light-activated bactericidal surface, while the pristine back side enables high-fidelity microbial capture for visual detection. The AIE-LGFM demonstrates outstanding field performance, accurately monitoring airborne microbes while enabling on-site photodynamic elimination. This work thus pioneers an eco-friendly and versatile membrane platform that unifies the all-in-one "sampling-detection-elimination" workflow, opening new avenues for intelligent management of airborne biological threats.

IntroductionNanozymes have emerged as promising substitutes for natural enzymes in chemiluminescent immunoassays, offering distinct catalytic advantages and superior stability. Despite their potential, many conventional nanozymes are constrained by a strong dependence on pH, which limits their effectiveness in certain assay environments. This highlights the need for nanozymes that maintain robust catalytic activity under alkaline conditions and are compatible with luminol-based detection systems.MethodsIn this study, we synthesized platinum nanoparticle-modified Prussian blue cubes (PB@Pt) and evaluated their enzyme-mimicking activity. The catalytic performance of PB@Pt was assessed under both weakly acidic and alkaline conditions. Its ability to enhance the luminol-H2O2 chemiluminescence (CL) system was investigated, and the CL signals were compared to those generated by natural horseradish peroxidase (HRP). Based on these properties, a novel CL immunoassay utilizing PB@Pt was developed for the sensitive detection of vascular endothelial growth factor (VEGF).Results and DiscussionThe synthesized PB@Pt exhibited high catalase (CAT)-like activity across a broad pH range, including alkaline media. Remarkably, in alkaline conditions, PB@Pt catalyzed the luminol-H2O2 reaction, producing CL signals significantly stronger than those achieved with natural HRP. Leveraging this enhanced performance, the established PB@Pt-based CL immunoassay enabled a wide linear detection range, ultra-low detection limits, high specificity, and excellent stability for VEGF quantification. This work introduces a novel strategy for designing CAT-mimicking nanozyme probes, thereby broadening their utility in CL immunoassays and advancing the clinical translation of nanozyme-based diagnostics for applications such as biomarker screening and point-of-care testing (POCT).

IntroductionVibrio cholerae relies on two-component signal transduction systems to adapt to the host intestinal microenvironment. This study investigated the role and regulatory mechanism of the BaeS/BaeR two-component system in intestinal colonization and antimicrobial peptide resistance.MethodsA baeS/baeR deletion mutant and complemented strain were constructed in the O1 El Tor clinical isolate EL2382. Intestinal colonization, Caco-2 cell adhesion, histopathological analysis, transcriptomic profiling, qRT-PCR, western blotting, electrophoretic mobility shift assays, chromatin immunoprecipitation-qPCR, and antimicrobial peptide resistance assays were performed.ResultsExpression of baeS and baeR was markedly induced during intestinal colonization and adhesion to Caco-2 cells. The ΔbaeS/baeR mutant exhibited significantly impaired bacterial adhesion and in vivo colonization, accompanied by reduced histopathological scores, and these phenotypes were restored by genetic complementation. Transcriptomic analysis identified salX (VC2553), which encodes an ABC transporter involved in antimicrobial peptide homeostasis, as a key downstream target of BaeS/BaeR. BaeR bound to the salX promoter and activated its transcription, and deletion-mapping electrophoretic mobility shift assays localized a putative BaeR-responsive sequence to 5′-TTCTTTTT-3′ within the −10/−35 spacer region. Similar to the ΔbaeS/baeR mutant, the salX mutant exhibited reduced resistance to human defensin 5 and impaired intestinal colonization. L-arginine exposure was associated with dose-dependent activation of the BaeS/BaeR pathway and BaeS/BaeR-dependent induction of salX, although a direct physical interaction between L-arginine and BaeS was not established.DiscussionThese findings identify an L-arginine-associated BaeS/BaeR–SalX regulatory pathway that promotes antimicrobial peptide resistance and intestinal colonization by V. cholerae, providing new insight into host cue-associated regulation of bacterial colonization fitness.

Proteolysis-targeting chimeras (PROTACs) have emerged as a transformative therapeutic strategy for targeted protein degradation, offering substantial potential for treating various diseases by specifically eliminating pathogenic proteins. However, the clinical translation of PROTAC technology faces significant challenges due to limited target selectivity, which may lead to off-target effects and potential toxicity in healthy tissues. Recent advances have focused on developing conditionally activatable PROTAC systems to improve their specificity and safety profile. This review comprehensively examines two primary classes of engineered PROTACs: (i) physically regulated systems, including photo-activatable, radiation, and ultrasound-activatable platforms, and (ii) chemically controlled approaches employing bioorthogonal conjugation strategies. We systematically analyze their molecular mechanisms, current clinical applications in oncology, and existing limitations regarding therapeutic efficacy and safety. The development of these precisely controlled PROTAC platforms promises to significantly enhance the clinical utility of targeted protein degradation technology in modern drug discovery.

BackgroundDoxorubicin (Dox)-induced cardiotoxicity (DIC) is initiated by acute stress that triggers early senescence-like phenotypes and severe metabolic maladaptation, yet effective therapeutic interventions remain elusive. Tricholoma matsutake polypeptide (TMP), a bioactive component from a medicinal fungus, exhibits antioxidant properties, but its potential to modulate cardiac energy metabolism remains unexplored.MethodsIn vivo, Sprague-Dawley rats were administered Dox (20 mg/kg, i.p.) with or without TMP (200/400 mg/kg/day) for 7 days. Cardiac function was assessed via high-resolution echocardiography, and metabolic signatures were decoded using LC-MS/MS-based untargeted metabolomics. In vitro, H9c2 cardiomyocytes were co-treated with Dox (5 μM) and TMP (30 μg/mL). Senescence phenotypes (SA-β-gal), mitochondrial dynamics (JC-1/Fura-2AM), and energy metabolites (ATP/NAD+) were quantified. Mechanistic axes were interrogated via Western blotting.ResultsTMP significantly attenuated Dox-induced cardiac dysfunction, interstitial fibrosis, and inflammation. Metabolomics revealed that TMP mitigated the glycolysis-dominant metabolic reprogramming, restored phospholipid homeostasis, and preserved the nucleotide pool. In cardiomyocytes, TMP antagonized premature senescence and suppressed ROS generation without compromising Dox’s antitumor efficacy in MCF-7 cells. Mechanistically, TMP alleviated the Dox-induced blockade of autophagic flux (restoring Beclin1/p-Parkin) and reactivated the AMPK/Sirt1/PGC-1α energy-sensing pathway, thereby preserving the ATP/NAD + reserve.ConclusionTMP functions as a metabolic modulator that protects against DIC by reprogramming energy metabolism and restoring mitochondrial quality control. These findings highlight TMP as a promising adjuvant for preserving cardiovascular health in cancer survivors.

Proteolysis-targeting chimeras (PROTACs) have revolutionized drug discovery by enabling targeted protein degradation via the ubiquitin-proteasome system (UPS), overcoming limitations of traditional occupancy-based inhibitors. This innovative approach has expanded the therapeutic landscape, particularly for previously “undruggable” targets. Recent advances have led to the emergence of dual/multi-target PROTACs, which simultaneously degrade multiple disease-associated proteins, offering a novel strategy to tackle complex pathologies driven by interconnected molecular networks. Such multi-targeting is especially relevant in multifactorial diseases, including cancer and neurodegenerative disorders, where single-target inhibition often proves insufficient. This review comprehensively examines the latest advancements in dual-target/multi-target PROTACs, including small-molecule PROTACs, biomacromolecule PROTACs, and nano-PROTACs, with a particular focus on the design principles, mechanism insights, and therapeutic potential in various diseases. Furthermore, we critically evaluate existing challenges-such as selectivity, pharmacokinetic optimization, and resistance mechanisms-and discuss future perspectives to maximize their clinical impact in precision medicine.

Osteoarthritis (OA) is a prevalent joint disorder characterized by progressive cartilage degradation, impaired mesenchymal stem cell (MSC) function, and chronic inflammation, ultimately leading to irreversible structural damage and functional impairment. Despite its high global burden, no regulatory agency has yet approved a disease-modifying therapy for OA, and effective interventions to halt or delay its progression remain a major challenge. Recent research highlights the pivotal role of the immune system in OA pathogenesis, with immunomodulatory biomaterials emerging as a promising strategy to simultaneously regulate inflammatory responses and promote tissue regeneration. These biomaterials, by leveraging their biocompatibility and immunoregulatory properties, offer a transformative alternative to conventional OA therapies, which predominantly focus on symptom management rather than targeting the underlying disease mechanisms. In this review, we comprehensively examine various immunomodulatory biomaterial strategies designed to mitigate OA progression. We first elucidate the immune landscape of OA, detailing the interplay between inflammation and disease pathophysiology. Next, we explore the latest advancements in immunomodulatory biomaterials, including nanoparticles (NPs), hydrogels, and scaffolds, highlighting their potential to reshape OA treatment. Finally, we discuss existing challenges and propose future directions for optimizing biomaterial-based immunotherapies to enhance OA management.

Post-operative surgical wound monitoring remains a significant clinical challenge in preventing bacterial infection. Current methods rely on indirect observations or costly investigations, often detecting infections only after complications arise. Here the medical sutures coated with Janus-type nanomotors (Pt-MOFs) with infected microenvironment-responsive properties for monitoring and treating surgical site infections are prepared. The Pt-MOFs nanomotors exhibit efficient self-propulsion with enhanced penetration and diffusion in biofilms by catalyzing hydrogen peroxide to produce oxygen bubbles. Copper ions serve dual roles as structural nodes and Fenton-like catalysts, generating antibacterial hydroxyl radicals while forming non-emissive self-aggregates. Here in vitro is shown that Pt-MOFs nanomotors present excellent bacterial imaging and enhanced antibacterial activity against both Gram-positive and Gram-negative bacteria. As a proof of concept, Pt-MOFs nanomotors coated surgical sutures successfully monitor the process of Staphylococcus aureus-infected wounds on mouse model. Furthermore, in vivo studies testify that Pt-MOFs nanomotors play an important role in treating infected surgical wounds through mitigating inflammatory infiltrates, facilitating collagen deposition and accelerating reepithelialization. This combined monitoring and treatment approach offers a promising strategy for surgical wound healing.

Rational capture of radioactive iodine is of great significance for nuclear energy development and environmental health. However, current nano-adsorbents suffer from limited adsorption capacity and frequent clogging induced by their powder form. In this work, we constructed a highly elastic nanofiber aerogel (MoS2@PPH-NFA), which assembled multiple available binding sites (amino-, carboxyl- and sulfide- groups) for synergistically and efficiently capturing molecular and ionic iodine. The aerogel possessed a hierarchically porous structure and exhibited outstanding mechanical integrity even after 20 compression cycles. Thanks to the synergistic effect between multiple adsorption sites, the aerogels could adsorb iodine vapor with a capacity of 4.02 g/g. Both experimental results and theoretical calculations verified that all the amino-, carboxyl- and sulfide- groups in aerogels demonstrated high binding affinity towards iodine species, wherein the amino ones played a predominant role with adsorption energy of-2.33 eV. MoS2@PPH-NFA also achieved ionic-iodine capture in aqueous with a maximum adsorption amount of 1.03 g/g, in which chemisorption dominated the adsorption process. In addition, the high flexibility and structural stability endowed MoS2@PPH-NFA with potential applicability for molecular/ionic iodine adsorption in mobile phases with more than 50 % removal rate. Overall, this study not only demonstrated the excellent performance of MoS2@PPH-NFA working as a promising iodine adsorbent but also provided a vivid prototype of developing novel fiber aerogels for environmental remediation.

Highly efficient detection and sterilization techniques for bioaerosol prevention and control are urgently needed. Herein, we present an AIE-active Janus air filter membrane (AIE-HAFM) that features water-dissolvable micro-nano porous network architecture and aggregation-induced emission (AIE) activity constructed by the asymmetrical surface modification with an amphiphilic AIE photosensitizer (MeOTTVP). The all-round AIE-HAFM can not only provide low pressure drop and high interception efficiency for bioaerosol sampling but also perfectly inherit the AIE functions of MeOTTVP, which allows for intensive near-infrared (NIR) emission and efficient production of reactive oxygen species. The airborne pathogens can be effectively captured, collected, transferred, and released by AIE-HAFM for subsequent quantitative detection with colony counting and ATP bioluminescence, as well as stained by the incorporated MeOTTVP for NIR fluorescence imaging-guided visual detection. Meanwhile, AIE-HAFM enables on-demand and surface-dependent photodynamic effects for reliable bacterial eradication under white light irradiation due to the surface-concentrated MeOTTVP, consequently achieving the smart prevention and control of bioaerosols both in the simulated and real-world bioaerosol environment. The versatility of AIE-HAFM in handling diverse airborne pathogens may bring about a transformative solution to address the bioaerosol contamination problems.

Mulberry (Morus nigra L.) is traditionally recognized for its bone-strengthening properties, yet its active constituents and underlying mechanisms remain incompletely understood. This study investigates the effects of maclurin, a key flavonoid from mulberry, on extracellular matrix (ECM) deposition and bone formation. In vitro, maclurin (5-20 μM) treatment enhanced ECM protein expression, including fibronectin (FN), laminin subunit gamma-2, and collagen alpha-1(I) chain (Col I) in MC3T3-E1 osteoblasts in a dose-dependent manner. Transcriptomic analysis suggested the involvement of PI3K/Akt signalling pathway, which was subsequently validated by increased Akt phosphorylation that was reversed by siAkt and PI3K inhibitor LY294002. In a subcutaneous ectopic bone formation model, oral maclurin administration (0.5-2.0 mg/kg) promoted Col I and FN expressions, improved collagen fibre organization, and increased calcium deposition in newly formed bone tissues. These results provide experimental evidence for osteogenic activity of maclurin and enhance our understanding of the bone-health benefits associated with mulberry.

Programmed death-ligand 1 (PD-L1) is a key immune checkpoint protein that enables tumor immune evasion by engaging PD-1 on T cells, thereby inhibiting their cytotoxic function. Although immune checkpoint inhibitors (ICIs) targeting the PD-1/PD-L1 axis have marked a breakthrough in cancer therapy, their effectiveness is often hampered by acquired resistance and immune-related adverse effects. Proteolysis-targeting chimeras (PROTACs) present a transformative approach by catalytically degrading PD-L1 instead of merely blocking its interaction with PD-1, offering a promising strategy to circumvent resistance and amplify therapeutic efficacy. This review systematically examines recent advancements in PD-L1-targeting PROTACs, including small-molecule PROTACs, biomacromolecule PROTACs, and nano-PROTACs, covering molecular design principles, mechanistic underpinnings, and preclinical validation. We also discussed how PD-L1 degradation can improve immunotherapy by regulating the tumor microenvironment (TME) and enhancing T-cell-mediated cytotoxicity. By synthesizing these insights, this review provides a roadmap for developing next-generation precision immunotherapies leveraging targeted protein degradation.

HER2 is a well-established oncogenic driver in breast, gastric, and other solid tumors. While HER2-targeted therapies such as trastuzumab and pertuzumab have improved clinical outcomes, resistance, particularly to trastuzumab, remains a major therapeutic challenge. Here, we engineered two IgG-VHH biparatopic antibodies (bpAbs), A9B5-Bs-5 and A9B5-Bs-7, incorporating an ECD I-binding nanobody A9B5 with the IgG scaffolds. These bpAbs target non-overlapping epitopes on the HER2 extracellular domain, promoting rapid receptor internalization and demonstrating superior antitumor activity compared to the trastuzumab and pertuzumab combination in trastuzumab-resistant tumor cells. Structural modeling suggests that both bpAbs engage HER2 in a trans-binding mode, leading to receptor clustering and interference with ligand-driven HER2 heterodimerization. These findings demonstrate that epitope-guided biparatopic antibody design can enhance HER2 downregulation and restore sensitivity to HER2-targeted therapy in vitro, providing a strategy for the development of next-generation receptor-targeted biologics.

Cell-culture room provides an interference-free clean environment for sterilized experimental operation and pharmaceutical production with high standards. However, the recurrent bioaerosol contamination poses a serious threat to its normal operation. Trapping by the low concentration of airborne microbes in such clean rooms, the sampling and characterization of bioaerosol contamination are appealing yet significantly challenging tasks. Herein, inspired by the integrative advantages of green dissolvable filter (HAFM), the bioaerosol particles in cell-culture rooms were captured and systematically investigated by cultivated and non-cultivated methods. Benefiting from the higher interception efficiency and superior compatibility than the commonly used sampling strategies, the HAFM-captured bioaerosol samples can optionally perform colony cultivation, counting, microscopic observation, and high-resolution imaging. Importantly, the real composition and abundance of the microbial community in cell-culture rooms can be presented by combining filtered sampling with high-throughput sequencing. The shared bacterial genera of Pseudomonas (44.34 %), Aquabacterium (14.25 %), Dechloromonas (3.17 %), Tepidiphilus (3.02 %), and Fluviicola (2.72 %), and fungal genera of Aspergillus (17.39 %), Cladosporium (12.62 %), Apiotrichum (8.91 %), Epicoccum (6.82 %), and Alternaria (5.02 %), are responsible for the bioaerosol contamination in cell-culture rooms. This work provides a promising avenue to overcome incompetence in accurate bioaerosol detection and identification in clean rooms.