Dysregulated tyrosine phosphorylation drives cancer proliferation directly. Though monoclonal antibodies (mAbs) and tyrosine kinase inhibitors (TKIs) regulate phosphorylation and suppress tumor growth, they are restricted for broader application because of side effects, including anaphylaxis and acquired resistance. Herein we propose a molecularly imprinted polymer@cationic liposome (MIP@Lip) that occupies a phosphorylated tyrosine epitope directly for cancer therapy. Human epidermal growth factor receptor 2 (HER2) was used as a model target in this work. After incubation with HER2+ cancer cells, MIP@Lip was internalized into lysosomes and subsequently escaped into the cytoplasm via the assistance of the cationic lipid. Subsequently, MIP@Lip directly occupied the phosphorylated tyrosine epitope of HER2, inhibiting its phosphorylation and downstream ERK signaling, suppressing over 80% of HER2+ cancer cell proliferation in vitro and tumor growth in vivo. Collectively, as phosphorylated tyrosine promotes proliferation signals directly, MIP@Lip is a "direct-occupancy-driven" strategy, and it offers a promising alternative to conventional targeted therapy.

Cordycepin (COR) is an important bioactive nucleoside with antitumor and immunomodulatory activities, but its conventional production via Cordyceps militaris fermentation is limited by long cultivation periods and low titers. Therefore, microbial heterologous synthesis represents an attractive alternative. In this study, the COR biosynthetic pathway was introduced into Saccharomyces cerevisiae CEN.PK530-1C, resulting in a COR titer of 870.7 mg/L. To further enhance COR biosynthesis, different fusion constructs were evaluated to relocalize the key enzymes into membraneless organelles in CEN.PK530-1C, which markedly increased COR production to 1209.5 mg/L. Metabolic engineering strategies were then applied to prevent COR degradation through deletion of the adenosine deaminase gene (YJL070C) and to enhance ATP and NADPH supply via overexpression of pgk1 and pos5, resulting in strain SCC7 with a COR titer of 1515.5 mg/L. Furthermore, metabolic effector molecules were screened and used to improve COR production to 1676.5 mg/L. In fed-batch fermentation, SCC7 achieved 6972.8 mg/L COR in a 6 L bioreactor, representing the highest COR titer reported in S. cerevisiae to date. In conclusion, this study establishes an efficient yeast platform for COR production and provides a useful reference for biosynthesis of other bioactive nucleosides.

Reg3A, a member of the regenerating islet-derived protein family, possesses both antibacterial and trophic effects, offering substantial benefits in the treatment of bacteria-induced wound infections. To overcome the challenges of protein drug inactivation and degradation in the hostile microenvironment of severely infected wounds, a multifunctional hydrogel dressing composed of polyvinyl alcohol (PVA), the bifunctional crosslinker TSPBA (TPA), and recombinant Reg3A protein (rReg3A) has been developed. The TPA-PVA/rReg3A hydrogel responds to infection-induced reactive oxygen species (ROS) through dynamic boronate ester bonds. Meanwhile, it releases the active rReg3A protein in a sustained manner, thereby delivering prolonged robust antibacterial and wound-healing properties. It was demonstrated that TPA-PVA/rReg3A hydrogel exhibited significant bactericidal activity against both Staphylococcus aureus (S. aureus) and methicillin-resistant Staphylococcus aureus (MRSA). In the MRSA-induced severe wound infection murine model, TPA-PVA/rReg3A hydrogel effectively promoted wound healing by clearing pathogenic bacteria, enhancing re-epithelialization, and stimulating hair follicle formation. As expected, the therapeutic effect of the TPA-PVA/rReg3A hydrogel was obviously better than that of the active rReg3A alone. In conclusion, our study provides a promising therapeutic strategy for managing severe wound infections by enhancing the bioactivity of antimicrobial proteins.

Molecular analyses of individual cells with high resolution, specificity, and sensitivity can not only reveal cellular heterogeneity but also provide a better understanding of diseases and accelerate drug discoveries. Single-cell endoscopy is an advanced live-cell technique that relies on a smart endoscope that allows minimally invasive probing of the interiors of individual cells. Compared with other single-cell analysis techniques, single-cell endoscopy has shown great promise in applications such as flexible single-cell manipulation, ultrasensitive sensing, and precise intracellular delivery. In this review, we aim to map out the landscape of recent advances in single-cell endoscopy techniques by focusing on both fundamental considerations and significant progress over the past decade. Specifically, we summarize the predominant live-cell endoscopes, including their fabrication and characterization. Furthermore, a series of valuable intracellular molecular sensing events, such as nucleic acids, proteins, ions, etc., are introduced with a main emphasis on how single-cell endoscopy can solve these issues and what merits single-cell endoscopy can provide. Finally, we briefly outline the remaining challenges and directions for the future development of single-cell endoscopy techniques.

Biofilms enhance microbial tolerance to harsh environments while preserving cellular activity over prolonged periods. Although biofilm-based continuous fermentation is widely applied for production of small-molecule chemicals, its application to macromolecular protein production has been rarely reported. Here, FimH, an adhesin from E. coli fimbriae that is employed for mannose-specific adherence and promotes biofilm formation, was displayed on the surface of S. cerevisiae using the anchoring proteins Sag1C, Sed1, Cwp2 and Ccw12, resulting in an 80-150 % enhancement in biofilm formation. Among them, Sed1 was the most effective, followed by Sag1C. A biofilm-based fermentation system for continuous secretion of human lysozyme (hLYZ) was established via cell-surface display of FimH in S. cerevisiae, achieving stable operation for over 350 h. The engineered strain BY4742-Sag1C-hlyz achieved an average extracellular hLYZ activity of 113.1 U/mL, significantly higher than the control BY4742-hlyz (41.6 U/mL), owing to enhanced cell adhesion that increased the total cell number in biofilm-based fermentation. Its productivity reached 2.36 U/mL/h, representing a 77.4 % increase compared with free-cell fermentation of BY4742-hlyz (1.33 U/mL/h). In conclusion, this study first achieved heterologous expression of a bacterial biofilm-forming gene in S. cerevisiae, enhancing biofilm formation and providing a reference for future expression of bacterial biofilm-related genes in yeast. Furthermore, biofilm-based fermentation enabled continuous secretion of hLYZ, highlighting a promising strategy for continuous production of recombinant proteins.

Interstitial skin fluid (ISF) holds great potential in bioassays as its components are similar to blood, offering a noninvasive alternative for disease diagnosis, prevention, and treatment. Microneedles (MNs), a novel biomedical tool, enable minimally invasive and painless extraction of ISF. This perspective systematically examines the latest advancements in MN-based ISF extraction and sensing. It elaborates on the differences between blood and ISF in molecular concentration and lag time, and compares MN-based sensing with conventional detection methods. Multiple biosensing principles, including electrochemical, fluorescence, colorimetry, and SERS, are detailed, and their metrics are evaluated in terms of LOD, specificity, response time, stability, and signal-to-noise ratio. The limitations of MNs, such as extraction efficiency, biological contamination, microdetection, and signal drift, are discussed, along with potential solutions. The application of MNs in wearable health monitoring, covering real-time monitoring, wireless data transmission, sensor miniaturization, closed-loop drug monitoring and delivery systems and commercialized MN detection products, is explored. Although challenges remain in sensor stability and device-clinical integration, the development of MN-and ISF-based detection devices is promising, and with further research, they are expected to revolutionize personal medical health monitoring.

Oxygen permeability is the primary requirement for mixed ionic electronic conductor (MIEC) membranes. Halogen doping enhances the permeability of MIEC membranes. Various halogens (F, Cl, and I) affect the performance of perovskite oxides differently, owing to variations in electronegativity, ionic radius, and other properties. In this work, an F and Cl co-doping strategy was proposed to improve perovskite oxygen permeation. A series of Ba0.5Sr0.5Co0.8Fe0.2O3-delta ClxF0.1-x materials were synthesized by the solid phase reaction method, and the impact of binary halogen doping on the crystal structure and oxygen permeability performance was systematically studied. Oxygen permeability tests suggested that the Ba0.5Sr0.5Co0.8Fe0.2O3-delta Cl0.06F0.04 disk membrane had good performance of 2.95 +/- 0.046 mL min- 1 & sdot;cm-2, confirming the benefits of the binary halogen- doped strategy. This improved performance results from the F and Cl co-doping, which enhances the oxygen vacancy concentration compared to Ba0.5Sr0.5Co0.8Fe0.2O3-delta materials, lowers the average metal-oxygen bond energy, and increases electrical conductivity. These results suggest that the binary halogen-doping strategy offers significant progress in the development of high-performance MIEC materials and potentially provides a new basis for the application of MIEC membranes.

Tailoring the magnetic properties and spectroscopic characteristics of transition metal (TM) clusters by doping non-metallic atoms is essential for the development of novel superatoms. Here, we employ density functional theory (DFT) to investigate the geometry, stability, electronic structure, and magnetic properties of the Co4Pn (n = 1-10) clusters. Our findings reveal that, except for Co4P, the clusters adopt a structure where phosphorus (P) atoms are added to the triangular faces of a Co4 core. These clusters exhibit high magnetic moments, primarily arising from the d-orbitals of Co atoms. Notably, Co4P4 adopts a highly symmetric Td point group structure, characterized by a core-shell configuration composed of Co4 and P4 tetrahedra. This structure, which represents the smallest cluster-based architecture, is confirmed through distance, charge, and orbital analyses, and is thermodynamically stable. Molecular orbital analysis uncovers a well-defined superatomic orbital arrangement of 1S2|1P6|2S2|1D10|2P6|1F14|3S2|2D8 with two electrons in parallel spin arrangements in the D orbitals, providing insights into the magnetic behavior of these superatoms. Furthermore, the infrared and Raman spectra of Co4P4 are further analyzed to establish a theoretical foundation for interpreting its electronic and geometrical properties. This study offers new perspectives on designing magnetic superatoms and tailoring their spectral response functions for targeted applications.

Cytoplasmic glutathione S-transferase (GST) is a key enzyme in cellular detoxification, catalysing the nucleophilic attack of glutathione (GSH) with toxic electrophilic substrates to produce less harmful compounds, thus aiding cellular detoxification. Studies have shown that GST is closely associated with the development of resistance to chemotherapeutic drugs, pesticides, herbicides and antibiotics, and the development of drug resistance in organisms poses new challenges in areas such as environmental protection and tumour therapy. In order to clarify the mechanism of GST in the development of drug resistance and detect the content of GST more accurately, this paper summarized the mechanism of GST on the development of drug resistance in different organisms, the types and research progress of organic small molecule fluorescence probes for GST imaging detection are introduced.

Moisture-electric generators (MEGs), which convert moisture potential into electrical energy, provide a promising alternative for energy generation due to the widespread and abundant distribution of air humidity in the surrounding environment. However, the low and intermittent electric output of MEGs presents challenges for their direct use in practical applications. In this study, an efficient hydrogel-based MEG integrated with hydrophilicity and ion concentration gradients based on a rational combination of polyvinyl alcohol, cellulose nanofibers, phytic acid, lithium bromide, and multi-walled carbon nanotubes was developed for the first time. The assembled MEG demonstrates outstanding power generation performance, with a single unit providing an output voltage of 1.0 V and a power density of 26.5 mu W cm- 2 (53.0 mu W cm- 3), enabled by the synergistic effect of its double-gradient structure. Notably, an increased current (4.0 mA) and voltage (45.3 V) were achieved using parallel and series integration of MEG banks, respectively. Scalable MEGs can directly power commercial electronic devices without the need for rectifiers or capacitors. This study provides new insights into MEGs design, contributing to the future development of sustainable power sources.

The advancement of nanomedicine requires a thorough understanding of the intrinsic bioactivity and molecular interactions of nanomaterials for safe and effective clinical applications, which remains lacking for most currently developed nanomaterials. Here, we uncover the unique intrinsic bioactivity and regulatory mechanisms of carbon-based fullerol nanomaterials through high-throughput molecular analysis and explore their therapeutic potential for tissue regeneration using tissue engineering approaches. Fullerol exhibits intrinsic pro-differentiation and antioxidant properties that enhance the osteogenesis and chondrogenesis of MSCs. Mechanistically, proteomic analysis combined with small-molecule inhibition studies reveals that fullerol is internalized by MSCs via clathrin-mediated endocytosis and activates NRF2 signaling, thereby exerting antioxidant effects that restore impaired MSC viability and differentiation under oxidative stress. Leveraging these unique bioactivities, we develop a fullerol-functionalized hydrogel with feasible physicochemical properties and triple biological functions in antioxidant, pro-osteogenic, and pro-chondrogenic effects and confirm its great regenerative capacity for both cartilage and subchondral bone by promoting structural restoration and improving functional recovery in a rat osteochondral defect model. Our findings offer new insights into the intricate interactions between stem cells and nanomaterials at the cellular and molecular levels and broaden the potential biomedical applications of fullerol for future cartilage and bone regeneration therapies.

Chronic diabetic wounds are characterized by a persistent inflammatory response, severe oxidative stress, and excessive proteolysis, creating an inhibitory microenvironment that impedes tissue regeneration. Recent findings indicate that regenerating family protein 3α (Reg3α) can promote keratinocyte proliferation and epidermal neogenesis, while also exhibiting antimicrobial properties. However, the low bioavailability significantly limits the clinical use of Reg3α in the treatment of chronic diabetic wounds. This study presents a glucose and ROS dual-responsive hydrogel loaded with Reg3α, which is synthesized by phenylboronic acid-modified hyaluronic acid (HAP) and polyvinyl alcohol (PVA). The Reg3α-loaded hydrogel (HAP-PVA/Reg3α), which exhibits favorable viscoelastic properties to adapt to wound application, promotes cell proliferation and demonstrates antibacterial and anti-inflammatory activities without inducing cytotoxicity or hemolysis in vitro. In diabetic mice, HAP-PVA/Reg3α effectively accelerates Staphylococcus aureus (S. aureus)-infected wound healing by alleviating bacterial infection, reducing inflammation, and facilitating collagen deposition. The result of RNA-seq suggests a negative regulation of M0 macrophages in the HAP-PVA/Reg3α group, which is presumably associated with their transformation into anti-inflammatory M2 macrophages. Meanwhile, serum pro-inflammatory IL-6 level is significantly decreased in Reg3α and HAP-PVA/Reg3α groups. In conclusion, HAP-PVA/Reg3α as a multifunctional hydrogel has significant potential for the treatment of chronic infected diabetic wounds.

The increasing presence of pharmaceutical active compounds (PhACs) in the environment has become a significant ecological and health concern, necessitating the development of efficient and cost-effective removal strategies. This study presents a novel composite, MIL-53(Al)@TA-Fe(III) MPNs, integrating metal-phenolic networks (MPNs) with metal-organic frameworks (MOFs) for efficient tetracycline removal from aqueous solutions. The composite was synthesized through an environmentally benign process, involving surface modification of MIL-53(Al) with tannic acid (TA) and Fe(III). Comprehensive characterization revealed that the optimal composite, achieved at a MIL-53(Al) to TA mass ratio of 1:1.2 and a TA to Fe(III) molar ratio of 1:5, exhibited a large specific surface area and pore volume, contributing to its high adsorption capacity of 532 mg/g for tetracycline. Adsorption isotherm, kinetics, and thermodynamic analyses indicated that the process was spontaneous, endothermic, and chemically driven. Mechanistic studies highlighted the roles of electrostatic and pi-pi interactions between the composite and tetracycline molecules. The composite also demonstrated excellent reusability, retaining over 92 % of its initial adsorption capacity after five cycles. This work not only provides a green and efficient strategy for tetracycline remediation but also offers valuable insights for the development of advanced adsorbents using MPNs-modified MOFs, holding significant potential for broader environmental applications in the future.

Background: Over the past 40 years since the discovery of regenerating family proteins (Reg proteins), numerous studies have highlighted their biological functions in promoting cell proliferation and resisting cell apoptosis, particularly in the regeneration and repair of pancreatic islets and exocrine glands. Successively, short peptides derived from Reg3δ and Reg3α have been employed in clinical trials, showing favorable therapeutic effects in patients with type I and type II diabetes. However, continued reports have been limited, presumably attributed to the potential side effects. Methods: This review summarizes extensive research on Reg proteins over the past decade, combined with our own related studies, proposing that Reg proteins exhibit dimorphic effects. Results: The activity of Reg proteins is not as simplistic as previously perceived but shows auto-immunogenicity depending on different pathophysiological microenvironments. The immunogenicity of Reg proteins could recruit immune cells leading to an anti-tumor effect. Such functional diversity is correlated with their structural characteristics: the N-terminal region contributes to autoantigenicity, while the C-type lectin fragment near the C-terminal determines the trophic action. It should be noted that B-cell masking antigens might also reside within the C-type lectin domain. Conclusions: Reg proteins have dual functional roles under various physiological and pathological conditions. These theoretical foundations facilitate the subsequent development of diagnostic reagents and therapeutic drugs targeting Reg proteins.

Precisely monitoring the alternations of biomolecules in traditional Chinese medicine-induced cells via a novel Raman fingerprint based analytical strategy.

Silicon inverted pyramids have been shown to exhibit superior SERS properties compared to ortho-pyramids, yet low-cost, simple preparation processes are lacking at present. This study demonstrates a simple method, silver-assisted chemical etching combined with PVP, to construct silicon inverted pyramids with a uniform size distribution. Two types of Si substrates for surface-enhanced Raman spectroscopy (SERS) were prepared via silver nanoparticles deposited on the silicon inverted pyramids by electroless deposition and radiofrequency sputtering, respectively. The experiments were conducted using rhodamine 6G (R6G), methylene blue (MB) and amoxicillin (AMX) molecules to test the SERS properties of the Si substrates with inverted pyramids. The results indicate that the SERS substrates show high sensitivity to detect the above molecules. In particular, the sensitivity and reproducibility of the SERS substrates with a denser silver nanoparticle distribution, prepared by radiofrequency sputtering, are significantly higher than those of the electroless deposited substrates to detect R6G molecules. This study sheds light on a potential low-cost and stable method for preparing silicon inverted pyramids, which is expected to replace the costly commercial Klarite SERS substrates.

Docetaxel (DOC) is commonly used in cancer treatment, especially for breast cancer. However, there are severe side effects in clinical application. In order to deliver docetaxel more effectively, a novel, active targeting acid-responsive polymer called cRGD-PAE-PEG-DSPE was developed. The polymer structure incorporated poly-(ethylene glycol) (PEG) as the hydrophilic segment, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) as the hydrophobic segment, and poly-(beta-amino ester) (PAE) as the acid-responsive group, which was grafted onto the PEG. Furthermore, c-(RGDyC) was grafted onto PAE to confer active targeting capability. Through self-assembly, docetaxel was encapsulated in RAED@DOC. Through in vitro experiments, it was confirmed that RAED@DOC had good serum stability and acid responsiveness, as well as enhanced uptake by MDA-MB-231 cells. Additionally, the antitumor efficiency in vivo and histopathological analysis showed that RAED@DOC exhibited higher antitumor activity and lower systemic toxicity in comparison to free docetaxel. These results suggested that RAED@DOC had considerable potential clinical use.

Atopic dermatitis (AD) is a chronic and recurrent inflammation disease associated with immune dysfunction. The high level of reactive oxygen species (ROS) causes high oxidative stress and further results in the deterioration of AD. At the same time, the ROS produced by bacterial infection can further aggravate AD. Here, the prepared PVA-based hydrogel (Gel) has a high ROS scavenging ability, and the antibacterial agent Zn-MOF(ZIF-8) loaded into the hydrogel shows a lasting and effective antibacterial activity. Thus, a Zn-MOF hydrogel (Gel@ZIF-8) is prepared to regulate ROS-mediated inflammatory microenvironment. In vitro experiments show that Gel@ZIF-8 has good antibacterial effect and cell biocompatibility. In the AD-induced mouse model, Gel@ZIF-8 can significantly enhance the therapeutic effect, such as reduce the thickness of epidermis, the number of mast cells and IgE antibodies. The results indicate that the ROS-scavenging hydrogel could treat the AD by regulating the inflammatory microenvironment, providing a promising treatment for managing AD.

Polycarbonate (PC) with high transmittance, stable mechanical performance and environmental resistance is crucial for practical applications. In this work, we report a method for the preparation of a robust antireflective (AR) coating by a simple dip-coating process of a mixed ethanol suspension consisting of tetraethoxysilane (TEOS) base-catalyzed silica nanoparticles (SNs) and acid-catalyzed silica sol (ACSS). ACSS greatly improved the adhesion and durability of the coating, and the AR coating exhibited high transmittance and mechanical stability. Water and hexamethyldisilazane (HMDS) vapor treatment were further employed to improve the hydrophobicity of the AR coating. The as-prepared coating exhibited excellent antireflective properties, with an average transmittance of 96.06% in the wavelength range of 400 to 1000 nm, which is 7.55% higher than the bare PC substrate. After sand and water droplet impact tests, the AR coating still maintained enhanced transmittance and hydrophobicity. Our method shows a potential application for the preparation of hydrophobic AR coatings on a PC substrate.

Dysfunction of intracellular proteins is frequently associated with various diseases, such as cancer. The exogenous proteins in cells are usually assembled with specific configurations due to physiological confinement/crowding to exhibit novel features in the protein structure, folding or conformational stability, distinguished with their behaviors in buffer solutions. Here, we synthesized exogenous proteins under confined/crowded conditions, to explore protein activity within cells. The findings suggested that the confinement and crowding effects on protein activity are heterogeneous; they showed an inhibitory effect on HRP by decreasing Km from ∼9.5- and ∼21.7-fold and Vmax from ∼6.8- and ∼20.2-fold lower than that of dilute solutions. Interestingly, the effects on Cyt C seem to be more complicated, and crowding exerts a positive effect by increasing Km ∼ 3.6-fold and Vmax ∼ 1.5-fold higher than that of dilute solutions; however, confinement exhibits a negative effect by decreasing Km ∼2.0 and Vmax ∼8.3 times. Additionally, in contrast to traditional nanoparticle-based confinement models, we synthesized a biodegradable nanoparticle to mimic the confined space, and the biggest advantage of this novel model is that the particles can be degraded and thus it can provide more intuitive observations of the properties of the target proteins under confinement and after release. Furthermore, we also evaluated protein activity in different cellular environments, indicating that the exogenous protein activity was closely related to the crowdedness of cellular environments, and the inhibition of protein activity in MDA-MB-231 cancer cells was more obvious than in HEK293 normal cells. Finally, SAXS analysis revealed the correlation between the protein conformation and the different environments. Our work will provide a unique method for precisely assessing whether the target cellular environments are native matrix in which specific exogenous protein drugs are delivered to function or whether they display a therapeutic role, which is of great significance for screening and development of new drugs.