As nanoscale metal-organic frameworks (MOFs) become increasingly prevalent, elucidating their fundamental interactions with proteins and cells is vital for evaluating their biological effects and biocompatibility. Herein, we investigated the protein corona (PC) formed on three representative MOFs (MIL-88, ZIF-8, UiO-66) and their impacts on macrophage uptake and cytotoxicity. Proteomic analysis revealed that the PC compositions were highly material-dependent, governed by the MOFs' distinct physicochemical properties. MIL-88 and ZIF-8 exhibited significant enrichment of acute-phase proteins and lipoproteins, whereas UiO-66 adsorbed high proportions of complement system proteins and lipoproteins. Apolipoprotein A-I (ApoA-I) was the most abundant protein across all three MOFs, while albumin was predominant on ZIF-8 and complement C3 was highly enriched on MIL-88. Functionally, PC formation reduced macrophage uptake of ZIF-8 and UiO-66 but had a negligible effect on MIL-88. The differences attributed to variations in dysopsonin/opsonin enrichment and surface charge alteration. Notably, the PC formation also significantly mitigated the cytotoxicity of ZIF-8 by reducing its cellular internalization. Together, these findings demonstrated that the PC is a pivotal factor influencing both the cellular interactions and the safety profile of MOFs, providing crucial insights for designing safer nanoparticle-based applications.

Phospholipids are biocompatible and versatile materials commonly used in the design of nanodelivery systems. However, the relationship between the structural characteristics of phospholipids and the physiological behavior of lipid-based nanoparticles remains inadequately understood. To explore the key structural features influencing efficient and targeted delivery, we created a library of phospholipid-coated gold nanoparticles (Lip@AuNPs), comprising 12 distinct formulations. These nanoparticles varied systematically in their headgroups (PA, PS, PC, and PE) and aliphatic chain lengths (6:0, 12:0, and 18:0). We investigated their effects on protein adsorption, cellular uptake, and in vivo delivery. Our findings showed that phospholipids with zwitterionic headgroups (PC and PE) reduced complement protein adsorption, enhanced selective uptake by nonphagocytic cells, and promoted increased accumulation in the spleen. Conversely, AuNPs coated with phospholipids containing shorter aliphatic chains exhibited higher serum protein adsorption, resulting in decreased and nonselective cellular uptake, which extended the circulation time of the nanoparticles in the bloodstream. This combinatorial approach provides valuable insights into the role of the phospholipid structure in nanoparticle design and offers practical guidance for developing lipid-based delivery systems with improved targeting and therapeutic efficacy.

Monitoring environmental drug-resistance genes (DRGs) plays a pivotal role in preventing the transmission of antimicrobial resistance, thereby reducing public health risks. In this study, a one-pot recombinase polymerase amplification (RPA)/clustered regularly interspaced short palindromic repeat (CRISPR) assay was developed for monitoring DRGs in river water. To overcome compatibility challenges between RPA and CRISPR systems, four glycosaminoglycans (heparin sodium, nadroparin calcium, dalteparin sodium, and chondroitin sulfate A sodium salt) with different molecular weights or negative charge density were evaluated as Cas-enzyme activity modulators. Among them, heparin sodium with the high molecular weight and high strong negative charge density exhibited the best performance in the one-pot DRG detection assay. In the system, CRISPR-Cas12a activity was temporarily inhibited during the RPA amplification phase. When sufficient amplicons were accumulated, Cas12a was activated for signal readout, thereby achieving orderly coupling and precise control of both reactions. To further simplify and improve the reliability of environmental DRG monitoring, a pretreatment method that can eliminate nucleic acid extraction was developed and integrated with the inhibitor-controlled one-pot platform. This assay achieved high sensitivity and specificity when it was applied to river samples, matching the performance of qPCR. The developed assay is simple to operate, has high sensitivity, and is widely adaptable, providing a robust tool for rapid antimicrobial resistance surveillance and exhibiting promise for public health management applications.

Vascular cell adhesion molecule-1 (VCAM-1) palys a key role in the occurrence and development of atherosclerosis (AS), making the monitoring and modulation of VCAM-1 expression critical for its diagnosis and treatment. To detect low-abundance VCAM-1 mRNA within macrophages, we have developed a highly sensitive ratiometric surface-enhanced Raman scattering (SERS) nanoprobe for the simultaneous precise detection and effective inhibition of VCAM-1 expression. This nanoprobe featured an Au-Au core-shell structure with dual functional layers: an interstitial 4-mercaptobenzonitrile (4-MBN) internal standard and an outer VCAM-1 mRNA recognition sequence tagged with Cy5. When the target mRNA bonded, it induced the formation of a stable duplex, distancing Cy5 from the metal surface and reducing its SERS intensity. While the shielded 4-MBN maintained a stable SERS signal. Therefore, the ultrasensitive detection of VCAM-1 mRNA within macrophages can be achieved by monitoring the proportional SERS signals of Cy5 and 4-MBN. Crucially, the binding event simultaneously suppressed VCAM-1 expression, enabling integrated diagnosis and pathological mitigation of AS. Under lipopolysaccharide (LPS)-induced pathological states, the nanoprobe dynamically tracked intracellular VCAM-1 mRNA levels while inhibiting mRNA/protein expression. This dual action blocked macrophage pyroptosis and attenuated inflammatory cytokine release, demonstrating therapeutic potential against AS progression. The ratiometric SERS nanoprobe design offers a versatile platform for nucleic acid-targeted theranostics in chronic diseases.

Pathogen nucleic acid analysis has emerged as an indispensable component of contemporary healthcare systems, serving dual roles in personalized clinical management and population-level disease surveillance. Herein, we present a novel G-quadruplex-integrated CRISPR-Cas biosensing platform that performs in a signal-on mechanism for colorimetric detection of pathogen nucleic acids in one-pot. By harnessing the unique properties of split G4 structures, we develop a universal visual probe that generates a distinct green colorimetric signal upon target recognition, which effectively couples with both Cas12 and Cas13 systems. Using the monkeypox virus (MPXV) B7R gene and a conserved respiratory syncytial virus (RSV) sequence as model targets for Cas12a and Cas13, respectively, we establish a detection workflow combining recombinase polymerase amplification (RPA) with CRISPR-mediated cleavage, visualized through enzymatic mediated color conversion. The tube-in-tube cartridge architecture adopted in this work enables seamless integration of RPA and CRISPR-based detection within a single closed-tube system, effectively eliminating cross-contamination risks. We successfully validate the platform for detection of MPXV in environmental samples and RSV in clinical specimens, achieving a detection limit of 1 copy per test and perfect concordance with PCR methods (40/40 agreement). The colorimetric biosensing platform developed herein demonstrates rapid (<60 min) and facial performance, establishing a novel molecular diagnostic paradigm that achieves laboratory-comparable accuracy for real-time surveillance and point-of-care applications.

Phenethyl isothiocyanate (PEITC) exerts anti-gastric cancer effect, however, the molecular mechanism has not yet been elucidated. KLF4 is an important regulatory molecule in gastric cancer. The programmed death ligand 1 (PD-L1) interacts with programmed cell death 1 (PD-1), inhibits function of cytotoxic T-lymphocyte and helps gastric cancer cells evade immune surveillance. The present study aimed to investigate the effect of PEITC against gastric cancer and the roles of KLF4 and PD-L1 in the anti-gastric cancer effect of PEITC. The effects of PEITC on gastric cancer cell proliferation and apoptosis were detected by EdU assay, flow cytometric analysis, immunoblotting and in vivo xenograft tumor experiment. The expression of KLF4 and PD-L1 was examined in PEITC-treated gastric cancer cells using immunoblotting, immunofluorescence and flow cytometric analysis. Co-culture system was used to assess the cancer cell-killing effect of PEITC combined with anti-PD-L1 blockade in gastric cancer. The results showed that PEITC suppressed cell activities both in vitro and in vivo. PEITC upregulated the expression of KLF4, which suppressed cyclin D1 expression and activated Bax expression, and mediated the growth inhibition effect of PEITC in gastric cancer. Meanwhile, we found that KLF4 induced by PEITC transcriptionally activated PD-L1 expression and diminished anti-cancer effects of anti-PD-L1 therapy in gastric cancer cells. Findings from this research revealed that PEITC restricted gastric cancer cell growth through activating KLF4. PEITC attenuated anti-PD-L1 therapy efficacy attributing to KLF4-upregulated PD-L1. These results uncover a novel mechanism of PEITC in gastric cancer intervention.

INTRODUCTION:Targeting immune checkpoint proteins (ICPs) via small molecules open a new window for cancer immunotherapy. Herein, we summarize recent advances of small molecules with novel chemical structures targeting ICPs, discusses their anti-tumor efficacies, which are important for the development of novel small molecules for cancer immunotherapy. AREAS COVERED:In this review, the latest patents and literature were gathered through the comprehensive searches in the databases of European Patent Office (EPO), Cortellis Drug Discovery Intelligence (CDDI), PubMed and Web of Science using ICPs and compounds as key words. EXPERT OPINION:To develop novel weapons to fight against cancer, small molecules targeting ICPs including CTLA-4, LAG-3, PD-L1, Siglec-9, TIM-3, TIGIT, and VISTA have been synthesized and evaluated in succession. Chief among them are the small molecules targeting PD-L1, which have been intensively investigated in recent years. Various in vitro assays such as ALPHA, HTRF binding assay, NFAT assay have been successfully developed to screen novel IPCs inhibitors. However, the in vivo assay, for example, using double-humanized PD-1/PD-L1 (hPD-1/hPD-L1) mouse as evaluation model, are seldom reported. Novel pharmacophores with new working mechanisms such as proteolysis targeting chimeras (PROTACs) and peptides are needed to enhance the therapeutic efficacy.

Simultaneous and multiplexed exosome protein profiling via an orthogonal CRISPR-Cas platform was achieved in this work. Aptamers were recruited to translate exosome surface protein information into Cas12a/Cas13a cleavage activity. The established multiplexed platform performed robustly with biological matrixes and could profile exosome proteins in clinical serum samples. AN orthogonal CRISPR-Cas platform is established for simultaneous and multiplexed circulating exosome phenotyping.

Single nucleotide polymorphisms (SNPs) are closely associated with many biological processes, including genetic disease, tumorigenesis, and drug metabolism. Accurate and efficient SNP determination has been proved pivotal in pharmacogenomics and diagnostics. Herein, a universal and high-fidelity genotyping platform is established based on the dual toeholds regulated Cas12a sensing methodology. Different from the conventional single stranded or double stranded activation mode, the dual toeholds regulated mode overcomes protospacer adjacent motif (PAM) limitation via cascade toehold mediated strand displacement reaction, which is highly universal and ultra-specific. To enhance the sensitivity for biological samples analysis, a modified isothermal recombinant polymerase amplification (RPA) strategy is developed via utilizing deoxythymidine substituted primer and uracil-DNA glycosylase (UDG) treatment, designated as RPA-UDG. The dsDNA products containing single stranded toehold domain generated in the RPA-UDG allow further incorporation with dual toeholds regulated Cas12a platform for high-fidelity human sample genotyping. We discriminate all the single-nucleotide polymorphisms of ApoE gene at rs429358 and rs7412 loci with human buccal swab samples with 100% accuracy. Furthermore, we engineer visual readout of genotyping results by exploiting commercial lateral flow strips, which opens new possibilities for field deployable implementation.

Environmental exposure to per- and poly-fluoroalkyl substances (PFAS) has raised significant global health concerns due to potential hazards in healthy adults. However, the impact of PFAS on susceptible populations, including pregnant individuals, newborns, the older people, and those with underlying health conditions, has been overlooked. These susceptible groups often have physiological changes that make them less resilient to the same exposures. Consequently, there is an urgent need for a comprehensive understanding of the health risks posed by PFAS exposure to these populations. In this review, we delve into the potential health risks of PFAS exposure in these susceptible populations. Equally important, we also examine and discuss the molecular mechanisms that underlie this susceptibility. These mechanisms include the induction of oxidative stress, disruption of the immune system, impairment of cellular metabolism, and alterations in gut microbiota, all of which contribute to the enhanced toxicity of PFAS in susceptible populations. Finally, we address the primary research challenges and unresolved issues that require further investigation. This discussion aims to foster research for a better understanding of how PFAS affect susceptible populations and to pave the way for strategies to minimize their adverse effects.

A split G-quadruplex based PfAgo sensing platform was developed for nucleotide mutation discrimination and human genotyping.

Rheumatoid arthritis (RA) is an autoimmune disease with multifactorial etiology and intricate pathogenesis. In RA, repeated monotherapy is frequently associated with inadequate efficacy, drug resistance, and severe side effects. Therefore, a shift has occurred in clinical practice toward combination therapy. However, conventional combination therapy encounters several hindrances, including low selectivity to arthritic joints, short half-lives, and varying pharmacokinetics among coupled drugs. Emerging nanotechnology offers an incomparable opportunity for developing advanced combination therapy against RA. First, it allows for co-delivering multiple drugs with augmented physicochemical properties, targeted delivery capabilities, and controlled release profiles. Second, it enables therapeutic nanomaterials development, thereby expanding combination regimens to include multifunctional nanomedicines. Lastly, it facilitates the construction of all-in-one nanoplatforms assembled with multiple modalities, such as phototherapy, sonodynamic therapy, and imaging. Thus, nanotechnology offers a promising solution to the current bottleneck in both RA treatment and diagnosis. This review summarizes the rationale, advantages, and recent advances in nano-empowered combination therapy for RA. It also discusses safety considerations, drug-drug interactions, and the potential for clinical translation. Additionally, it provides design tips and an outlook on future developments in nano-empowered combination therapy. The objective of this review is to achieve a comprehensive understanding of the mechanisms underlying combination therapy for RA and unlock the maximum potential of nanotechnology, thereby facilitating the smooth transition of research findings from the laboratory to clinical practice.

Autophagy, vital for removing cellular waste, is triggered differently by small molecules and nanoparticles. Small molecules, like rapamycin, non-selectively activate autophagy by inhibiting the mTOR pathway, which is essential for cell regulation. This can clear damaged components but may cause cytotoxicity with prolonged use. Nanoparticles, however, induce autophagy, often causing oxidative stress, through broader cellular interactions and can lead to a targeted form known as "xenophagy." Their impact varies with their properties but can be harnessed therapeutically. In this review, the autophagy induced by nanoparticles is explored and small molecules across four dimensions: the mechanisms behind autophagy induction, the outcomes of such induction, the toxicological effects on cellular autophagy, and the therapeutic potential of employing autophagy triggered by nanoparticles or small molecules. Although small molecules and nanoparticles each induce autophagy through different pathways and lead to diverse effects, both represent invaluable tools in cell biology, nanomedicine, and drug discovery, offering unique insights and therapeutic opportunities. Small molecules engage with molecular targets to clear damaged cellular components through non-selective autophagy, a process that can occasionally result in cytotoxicity. In contrast, nanoparticles non-selectively interact with cellular molecules to initiate autophagy or xenophagy that is impacted by their physicochemical properties. image

Clinical treatment and preclinical studies have highlighted the role of immune checkpoint blockade in cancer treatment. Research has been devoted to developing immune checkpoint inhibitors in combination with other drugs to achieve better efficacy or reduce adverse effects. Phytochemicals sourced from vegetables and fruits have demonstrated antiproliferative, proapoptotic, anti-migratory, and antiangiogenic effects against several cancers. Phytochemicals also modulate the tumor microenvironment such as T cells, regulatory T cells, and cytokines. Recently, several phytochemicals have been reported to modulate immune checkpoint proteins in in vivo or in vitro models. Phytochemicals decreased programmed cell death ligand-1 expression and synergized programmed cell death receptor 1 (PD-1) monoclonal antibody to suppress tumor growth. Combined administration of phytochemicals and PD-1 monoclonal antibody enhanced the tumor growth inhibition as well as CD4(+)/CD8(+) T-cell infiltration. In this review, we discuss immune checkpoint molecules as potential therapeutic targets of cancers. We further assess the impact of phytochemicals including carotenoids, polyphenols, saponins, and organosulfur compounds on cancer PD-1/programmed cell death ligand-1 immune checkpoint molecules and document their combination effects with immune checkpoint inhibitors on various malignancies.

Tetracycline (TC) is widely used in veterinary medicine and animal feed; however, TC residues in food pose a risk to human health. Thus, the sensitive and selective detection of TC is needed to ensure food safety. Herein, we developed a CRISPR-Cas12a biosensor with competitive aptamer binding to detect TC residues. The aptasensor, formed by hybridizing activator DNA with TC-specific aptamers on streptavidin-modified magnetic beads, releases activator DNA in a TC concentration-dependent manner. This activated the Cas12a-crRNA complex, which cleaved single-strand DNA reporters to generate a detectable fluorescence signal. The TC signal was amplified through a two-step incubation reaction, with a detection limit as low as 9.45 × 10-5 μg L-1. The assay showed high selectivity and good recovery rates in various biological samples (e.g., honey, milk, fish), demonstrating the applicability of the biosensors in pollutant detection.

Understanding protein corona composition is essential for evaluating their potential applications in biomedicine. Relative protein abundance (RPA), accounting for the total proteins in the corona, is an important parameter for describing the protein corona. For the first time, we comprehensively predicted the RPA of multiple proteins on the protein corona. First, we used multiple machine learning algorithms to predict whether a protein adsorbs to a nanoparticle, which is dichotomous prediction. Then, we selected the top 3 performing machine learning algorithms in dichotomous prediction to predict the specific value of RPA, which is regression prediction. Meanwhile, we analyzed the advantages and disadvantages of different machine learning algorithms for RPA prediction through interpretable analysis. Finally, we mined important features about the RPA prediction, which provided effective suggestions for the preliminary design of protein corona. The service for the prediction of RPA is available at http://www.bioai-lab.com/PC_ML .

The advent of drug-eluting contact lenses (DECLs) has opened up new avenues for the treatment of eye diseases. DECLs is expected to partially overcome the shortcomings of eye drops due to single-dose packaging, accurate dosing, prolonged drug elution behavior, and simplified dosing procedures. Currently, a significant proportion of the DECLs design effort has been directed towards enhancing the compatibility of contact lenses with drugs. The appropriate elution time for the drug remains unclear. Additionally, it is ambiguous for which ophthalmic diseases DECLs offers the greatest therapeutic advantage. To rationally design DECLs in practice, it is necessary to understand the acceptance of DECLs by patients and practitioners and to clarify the indications for DECLs. This review will first focus on the acceptance of DECLs by different patients and practitioners and discuss the factors that influence its acceptance. Secondly, this review presents an overview of the current effectiveness of DECLs treatments in animals and in the clinical phase, with a particular focus on the suitability of DECLs for the treatment of ophthalmic diseases. Overall, patients and practitioners expressed positive attitudes towards DECLs. However, this is related to factors such as DECLs’ treatment cycle, safety, and price. In addition, DECLs has good application prospects for ocular wound healing, postoperative management, and treatment of contact lenses-related complications. Furthermore, chronic diseases such as glaucoma that necessitate long-term medication and intraocular diseases that require implants or injections represent additional potential applications for DECLs. It is hoped that this review will facilitate a deeper understanding of DECLs acceptance and indications, thereby supporting the rational design of DECLs. At the same time, this review provides a reference for the design of other drug-device combination products.

Introduction: China implemented a dynamic zero-COVID strategy to curb viral transmission in response to the coronavirus disease 2019 (COVID-19) pandemic. This strategy was designed to inhibit mutation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for COVID-19. This study explores the dynamics of viral evolution under stringent non-pharmaceutical interventions (NPIs) through real-world observations. Methodology: Our analysis encompassed all polymerase chain reaction (PCR)-confirmed COVID-19 cases (n = 780) from two consecutive outbreaks occurring in July to August 2021. Epidemiological investigations and in-depth viral genome analyses were conducted to determine the sources of infection and the inter-outbreak linkages. Results: The outbreaks likely originated via contaminated objects from an imported COVID-19 case to local airport cleaners. Most patients presented clinically mild symptoms. All viral genomes clustered into a single clade, with genomes from the subsequent Yangzhou outbreak forming a distinct branch, diverging from those of the initial Nanjing outbreak. Compared to the Wuhan-Hu-1 strain, the Delta variants isolated from the imported case and early local cases in Nanjing had 35 nucleotide mutations and three deletions. Various single nucleotide variant (SNV) combinations were observed in subsequent transmissions, with NPIs accounting for the termination of 67.2% (92/137) of SNV combinations. Stringent implementation of NPIs resulted in a greater number of terminated SNV combinations than emerging ones. Additionally, vaccination of cases was associated with SNV combination termination. Conclusions: This study underscores the significant role of NPIs in preventing the accumulation of SARS-CoV-2 mutations.