Oxaliplatin (OXA) is a third-generation platinum-based chemotherapeutic drug for cancer therapy. However, OXA frequently induces peripheral neuropathy, especially cold allodynia, while the underlying mechanisms remain elusive. Herein, we notice that liposomal OXA (Lip-OXA) alleviates cold allodynia in mice compared with free OXA, despite comparable platinum accumulation levels in neural tissues in these two groups, suggesting indirect pathways stimulate peripheral nerves during the OXA therapy. Further investigation indicates that platinum accumulation in dermal extracellular matrix-associated fibroblasts activates the nuclear factor κB (NF-κB) pathway, followed by activation of the chemokine (C-C motif) ligand 2 (CCL2)/C-C motif chemokine receptor 2 (CCR2)/transforming receptor potential cation channel subfamily M member 8 (TRPM8) axis to induce cold allodynia. Nanoformulated OXA treatment reduces cold allodynia due to lower platinum accumulation in the dermis (than that of free OXA). Notably, Lip-OXA shows a superior antitumor efficacy over free OXA in an in situ colorectal cancer model. These findings expand the mechanisms underlying OXA-induced cold allodynia and suggest that nanoformulated OXA can effectively alleviate chemotherapy-induced peripheral neuropathy, while enhancing therapeutic efficacy.

In recent years, with the increasing incidence of other non-EV-A71 enteroviruses, the research on other enteroviruses has become more and more important. Coxsackievirus A14 (CVA14), A member of the enterovirus group A, causes hand-foot-mouth disease (HFMD), aseptic meningitis and acute abdominal pain. In this study, we established the first ICR mouse model of CVA14 infection. The CVA14 HE27 strain was selected to infect neonatal mice through a preliminary experiment, and the infection model was established according to three different conditions of infection dose, route and age. It was found that 2-day-old ICR neonatal mice showed clinical symptoms such as hind limb paralysis and death after intramuscular injection of 105.75 TCID50 CVA14 HE27 strain. Through daily monitoring, we found that the viral load of skeletal muscle was the highest, and there were certain pathological changes in various tissues and organs, among which the changes of brain and skeletal muscle were the most obvious. The expression of pro-inflammatory cytokines IL-4, IL-10, IFN-γ and TNF-α were abnormally high. This neonatal mouse model of CVA14 infection will be useful for the development of prophylactic and therapeutic multivalent vaccines, as well as for the screening of antiviral drugs against enterovirus.

Esophageal cancer is a malignant tumor with high morbidity and mortality. Cisplatin is a first-line chemotherapeutic drug for the treatment of esophageal cancer. However, nearly half of the patients eventually develop acquired resistance, making cisplatin resistance a critical clinical challenge in treating esophageal cancer. Cancer-associated fibroblasts (CAFs) play a critical role in cisplatin resistance of esophageal cancer. We found that primary CAFs isolated from cisplatin-resistant esophageal cancer patients exhibited significantly elevated S100A4 protein levels and enhanced resistance to cisplatin. Thus, the high expression of S100A4 in CAFs is suggested to be closely linked to cisplatin resistance in clinical esophageal cancer. Herein, we designed a CREKA-Lipid@Fe-siS100A4 (C-L@F-siA4) system that can efficiently target and eliminate the S100A4 gene within CAFs. In vitro and in vivo experiments demonstrated the ability of C-L@F-siA4 to reverse the resistance and restore the effectiveness of cisplatin in treatment of esophageal cancer. This study provides a target for treating esophageal cancer with acquired cisplatin resistance and offers a strategy as an adjuvant method for chemotherapy of esophageal cancer.

Coxsackievirus B1 (CV-B1) is primarily associated with meningitis but can also cause localized outbreaks of hand, foot, and mouth disease (HFMD). This study analyzed the genetic diversity of the VP1 gene in 39 strains of the CVB1 virus isolated from HFMD children across 15 provinces in China between 2010 and 2024, as well as 179 strains from 17 countries. Based on the average nucleotide difference of VP1 gene, we classified CVB1 virus into six genotypes A to F, Notably, genotype F is newly classified. Since 2010, genotype F guadually replaced genotype E as the dominant genotype in China and has further subdivided into three subtypes: F1, F2, and F3, with F3 being the most prevalent subtype in China currently. We specifically study the mild and severe cases within the F3 subtype. Temperature-sensitivity experiments revealed no differences between mild and severe cases of the F3 subtype, and they all belong to temperature-sensitive strains. Interestingly, we found that mild cases of the F3 subtype did not involve recombination, whereas all severe cases of the F3 subtype showed recombination with Coxsackievirus B4 (CVB4). CVB4 has consistently been the primary pathogen responsible for severe neonatal illnesses, suggesting that recombination between the F3 subtype and CVB4 may be associated with the development of severe HFMD. These findings provide fundamental scientific data for further investigation into the epidemiology and genetic characteristics of variants of Coxsackievirus B1 in China.

The immune system is essential for maintaining physiological balance, defending against pathogens, and eliminating abnormal cells. Immune dysregulation—manifesting as either suppression or hyperactivation—plays a central role in the pathogenesis of diseases such as cancer, rheumatoid arthritis, and chronic inflammatory disorders. As a novel class of catalytic nanomaterials, nanozymes possess enzyme-mimicking activities and exhibit high stability, biocompatibility, and tunable functionalities, enabling them to serve as versatile tools in immunomodulation. This review highlights the dual immunoregulatory capabilities of nanozymes and their context-dependent therapeutic applications, including promoting antitumor immunity, restoring immune homeostasis in autoimmune diseases, and attenuating pathological inflammation. We systematically discuss the mechanisms by which nanozymes modulate immune responses, reshape immune microenvironments, and influence key signaling pathways. These insights aim to facilitate the rational design of next-generation nanozyme-based immunotherapeutics.

Cholangiocarcinoma (CCA), the second most prevalent primary hepatic malignancy, demonstrates resistance to antiangiogenic therapy due possibly to the dynamic interaction between cancer-associated fibroblast (CAF)-mediated extracellular matrix (ECM) remodeling and angiogenesis. This study shows that anti-VEGFR2 therapy activates CAF, inducing excessive ECM deposition and forming a physical barrier that diminishes the effectiveness of antiangiogenic therapy in CCA. Based on the finding, we rationally engineered vascular endothelial cell-derived nanovesicles that inherit the angiogenic factor receptor that competitively bind and neutralize pro-angiogenic ligands to diminish their bioeffects. These nanovesicles also retain high levels of integrin αvβ3 and specifically carry peptide B7-33 (an inducer of fibroblasts quiescence) modified by the cRGD peptide, thereby developing dual-functional nanovesicles (B7-33-SNPs). The study revealed that B7-33-SNPs synergistically disrupted the CAF-angiogenesis crosstalk, effectively reducing microvessel density and fibrotic deposition in subcutaneous xenograft CCA models. This combinatorial strategy achieved a 67.7 ± 17.6

Portal vein tumor thrombus (PVTT) is a common and severe indicator in advanced hepatocellular carcinoma (HCC), characterized by a poor prognosis and limited response to existing therapies. Cancer-associated fibroblasts (CAFs) play an important role in promoting HCC metastasis and contribute to resistance against sorafenib (SOR) resistance, which is a standard treatment for advanced HCC. The data from single-cell RNA sequencing highlights the critical role of C-X-C motif chemokine ligand 12 (CXCL12) in the activation of CAFs. To address these challenges, we develop a PVTT-targeted nanocarrier designed to co-deliver small interfering RNA (siRNA) and a multikinase inhibitor, aiming to enhance therapeutic outcomes for PVTT. This novel lipid-coated polylactide-co-glycolide nanoparticle system effectively downregulate CXCL12 expression in CAFs, leading to their inactivation and subsequent reshaping of the tumor microenvironment. The resulting modulation of the tumor microenvironment significantly suppress tumor cell migration, invasion, and resistance to SOR, thereby demonstrating potent anti-tumor effects in orthotopic mouse models of PVTT. Furthermore, RNA sequencing reveals key regulatory pathways and genes associated with the inhibition of SOR resistance and PVTT formation mediated by these nanoparticles. These findings suggest that modulating the tumor microenvironment, combined with targeted anti-tumor therapies, offers a promising strategy for treating HCC patients with PVTT.

Nucleotide (nt) deletions in the VP1 region of poliovirus are extremely rare. However, as early as 2012, we detected a natural type 2 poliovirus strain in sewage, originating from the Sabin 2, which exhibited such deletions. Whole-genome analysis revealed that the virus genome is 7,436 nt in length, with three nucleotide deletions in positions 16-18 of the VP1 region (2,497-2,499 nt), resulting in a deletion of the amino acid at position six of the VP1 capsid protein. Notaly, the missing amino acids are not located at known attenuation or neutralization sites. In addition, the important attenuated sites at positions 481 and 2,909 remained unchanged. Only one substitution was observed at the known neutralizing antigen site: VP3-61 (Arg to Lys) on NAg3a. Recombinant analysis showed that the virus is a type 2/3 recombinant virus, with the crossover site located between nucleotides 6,981 and 7,439, spanning the 3Dpol region and the 3' untranslated region (UTR). The protein structure simulation showed that VP1 capsid protein of the PV-2 deletion variant was very similar to Sabin 2. In the VP1 region, the PV-2 deletion variant has only a triple nucleotide deletion, with no other mutations, indicates that the virus was in an early stage of evolution before being isolated from sewage. Furthermore, this variant is only a type 2/3 recombinant, not recombined with other non-polio enteroviruses, suggesting a short transmission and circulation time of the virus in the population. Therefore, we speculate that the nucleotide deletion in VP1 region of poliovirus may occur in the early evolutionary stage. Although the virus has not yet spread widely among humans, these findings highlight the importance of continuous environmental monitoring of poliovirus.

Coxsackievirus A10 (CVA10) has been reported frequently in many infectious diseases and cases associated with hand, foot, and mouth disease (HFMD) emerging increasingly in recent years. Based on the National HFMD System Surveillance, 180 CVA10 strains were isolated from the mainland of China between 2008 and 2023. These strains were analyzed alongside 103 representative full VP1 sequences obtained from GenBank, with a focus on global-scale phylogenetic analysis and spatiotemporal dynamics of CVA10. Eight genotypes (A-H) were defined, of which the genotype C was the dominant gene subtype in Chinese mainland. Bayesian analysis indicated that the most renascent common ancestor (tMRCA) of CVA10 originated in 1932 (95% HPD:1867-1958), with a high evolutionary rate of 3.32 × 10-3 substitutions/site/year (95% HPD: 2.62 × 10-3 to 3.40 × 10-3). By analyzing the spatial propagation paths, the global CVA10 exhibited distinct regional characteristics. Though the origin of CVA10 could be in the USA, regional dissemination was mainly located around the Asia-Europe region. The spatiotemporal dynamics of CVA10 exhibited frequent viral traffic among localities, and virus from East and South China have played a central role in spreading around the mainland of China. Our phylogenetic description and phylogeographic analyses indicate the importance of large spatial- and temporal-scale studies in understanding epidemiological dynamics of CVA10, particularly the diffusion routes will be of great importance to global control efforts.

ABSTRACT Poliovirus is characterized by three antigenically distinct serotypes that do not elicit cross‐neutralizing antibodies. In the final stages of poliovirus eradication, the gold‐standard conventional neutralization test (cNT) for detecting serum neutralizing antibodies (NAbs) is highly restricted due to biosafety concerns. To address this, we developed a high‐throughput, tri‐color pseudovirus‐based neutralization assay (PBNA) for the simultaneous quantification of NAbs against all three poliovirus serotypes. We generated pseudoviruses by co‐transfecting cells with P1 plasmids, a replication plasmid, and a T7 RNA polymerase plasmid. By optimizing P1 expression, sensitive cell selection (HEK 293T), and plasmid transfection ratios (3:3:1 for P1, replicon, and T7 plasmids), we produced high‐titer pseudoviruses (>29‐fold increase in titers). Based on high‐titer pseudovirus encoding distinct fluorophores (E2, eGFP, and RFP), the PBNA was established, which was optimized for a 12 h incubation period, 4 × 10⁴ cells per well, and 1500 TCID50/mL of pseudovirus. It demonstrated high sensitivity, strong serotype specificity, and excellent reproducibility. Furthermore, the PBNA and cNT exhibited excellent congruency (r > 0.88, all serotypes). The tri‐color PBNA provides a safe, rapid, and alternative to the cNT, making it an invaluable tool for large‐scale serosurveillance, novel vaccine evaluation, and fundamental virological investigations in the post‐eradication era.

BackgroundNeoadjuvant therapy (NAT) has transformed cancer treatment by improving surgical outcomes and survival rates, yet resistance mechanisms across multiple cancer types remain unclear. This study aimed to decipher tumor ecosystem dynamics during NAT using cross-cancer single-cell sequencing data, focusing on identifying key mediators of immunosuppression and treatment resistance.MethodsSingle-cell RNA-sequencing (scRNA-seq) datasets from five solid tumors (esophageal squamous cell carcinoma, esophagogastric junction carcinoma, colorectal cancer, cervical cancer, and triple-negative breast cancer) were integrated. The data from these five cancer types underwent a rigorous process to standardize cell types across all datasets. Cell-cell communication analysis, Meta-Programs (MPs) via non-negative matrix factorization, and functional enrichment were performed. Immunohistochemistry (IHC) and Western blot validated S100A4 expression and PD-L1 induction in vitro.ResultsWe constructed a single-cell map across cancer types and systematically characterized dynamic changes in tumor cells and diverse microenvironmental cell populations following neoadjuvant therapy, along with thier gene expression and pathway alterations. Our findings highlight that crosstalk between cancer-associated fibroblasts (CAFs) and tumor cells represents a critical determinant of neoadjuvant therapy resistance. Fibroblasts underwent significant state transitions post-treatment, marked by hypoxia-associated gene upregulation (e.g., S100A4) and immunosuppressive pathways. Meta-Programs (MPs) analysis identified a hypoxia-driven fibroblast state (MP5) containing S100A4 that correlated with treatment resistance. In vitro experiments, S100A4 co-localized with α-SMA + fibroblasts and directly induced PD-L1 expression in tumor cells, linking CAFs secreted S100A4 to immunosuppressive PD-L1 upregulation.ConclusionThis cross-cancer single-cell atlas reveals S100A4, secreted by CAFs, as a conserved mediator of PD-L1 upregulation in tumor cells, driving immunosuppression and resistance to nICT. The atlas and mechanistic findings provide a rationale for targeting S100A4 to enhance treatment efficacy, pending validation in larger cohorts and mechanistic studies. This resource also supports the development of personalized, cross-cancer neoadjuvant strategies.

Enterovirus D68 (EV-D68), a serotype of the enterovirus species D, has garnered significant attention due to outbreaks reported in 2014, 2016, and 2018. In this study, 36 Chinese EV-D68 strains were isolated, sequenced, and combined with all EV-D68 VP1 sequences from GenBank to form a data set of 1679 sequences. This data set served as the basis for phylogenetic, evolutionary dynamics, phylogeographic, and key amino acid site mutation analyses of EV-D68. Based on the VP1 region, EV-D68 is classified into four genotypes (A-D), and seven subgenotypes (B1-B3, D1-D4), with B3 and D3 being the predominant subgenotypes. Bayesian skyline plots indicated that genotypes B and D experienced multiple population expansions, aligning with reported EV-D68 outbreaks. Phylogeographic analyses of the B3 subgenotypes revealed sequences from Europe and North America clustering into a single evolutionary branch, suggesting significant transmission between these regions. Additionally, mutation analysis identified VP1-98 as a high-frequency mutation site, differing significantly between the previously prevalent A and C genotypes and the currently prevalent B and D genotypes. However, the functional implications of this mutation require further investigation. This study provides a solid theoretical basis for epidemiological research, disease surveillance, and prevention efforts related to EV-D68.

Coxsackievirus A10 (CVA10) is an enterovirus that has caused global outbreaks of hand, foot, and mouth disease (HFMD), accompanied by neurological and systemic complications. Animal models that can simulate natural infections are necessary for studying viral pathogenesis. In this study, our objective was to establish a mouse-adapted strain (CVA10-P8) through serial passaging, which was capable of orally infecting 14-day-old ICR mice, leading to hind-limb paralysis and death. Viral titers in various tissues indicated tropism in muscle tissue, with significant increases observed in the brain, lung, and intestinal tissues as the infection progressed. Pathological examination revealed tissue damage in the muscles, brain, lungs, and intestines accompanied by neutrophil infiltration of the brain. Furthermore, flow cytometry and transcriptome analysis revealed metabolic abnormalities, immune system activation, and the promotion of systemic inflammatory responses in the brain, muscle, and lung tissues. In summary, we successfully developed a CVA10 mouse-adapted strain and a corresponding mouse model, providing valuable tools for studying CVA10 pathogenesis and evaluating the efficacy of antiviral interventions.IMPORTANCECVA10 has emerged as a predominant pathogen in the etiology of HFMD, with the potential to elicit neurological manifestations and systemic complications. In this study, we successfully established a novel murine model of CVA10 infection by serially propagating a clinical isolate of CVA10, which enabled oral infection in 14-day-old ICR mice. This model facilitated the investigation of the pathogenesis of CVA10-induced disease. Utilizing this infection model, we employed flow cytometry and transcriptome analysis to elucidate the central nervous system (CNS) inflammatory responses elicited by CVA10 in mice, which closely mimic the natural route of infection. Our findings provide novel insights into the pathophysiological mechanisms underlying CVA10-induced neuroinflammation and pave the way for further research into targeted therapeutic interventions for HFMD associated with CVA10.

Regenerating functional bone tissue in critical-sized defects remains a formidable issue. Bone-tissue engineering (BTE) scaffolds are emerging as potential alternatives to bone transplantation for the repair of bone defects. However, developing BTE scaffolds with unique bone-healing properties and natural bone porous structure is challenging. Herein, we presented a biomimetic scaffold with hierarchical porosity via a solvent casting/particulate leaching method. The scaffold comprises osteoinductive whitlockite (WH) nanoparticles evenly dispersed in a poly(lactic-co-glycolic acid) (PLGA) matrix. Highly interconnected pores with hierarchical variations are present in the scaffold, enabling superior solution diffusion and compressive strength. Notably, the WH/PLGA scaffold effectively promoted osteoblast differentiation in vitro and induced bone formation in rat tibia defects, surpassing the performance of both the hydroxyapatite (HAP)/PLGA scaffold and the PLGA scaffold. This study provides a low-cost, facile, and scalable strategy for fabricating BTE scaffolds with favorable mechanical properties, biocompatibility, and bone repair capability.

Osteoarthritis (OA) is the most prevalent degenerative joint disease marked by cartilage degeneration, synovial inflammation and pain, which seriously affects life quality of patients. However, due to unclear pathological mechanisms, there is still lack of specific targets at the molecular level for OA treatment. Since OA-related cartilage displays pathological features of fibrosis, such as increased secretion of collagen I (COL I) but decreased secretion of collagen II (COL II), and cartilage fibrosis is usually defined as a final-stage of OA, we hypothesized that fibrosis related factors could promote OA progression. By combining public databases with pathological analysis of clinical OA immunohistochemistry samples, we found that a key pro-fibrotic factor, S100A4, also named fibroblast specific protein-1 (FSP-1), was overexpressed by OA chondrocytes and positively related with OA progression. To investigate if S100A4 can be a therapeutic target of OA, we designed cartilage-targeting lipid nanoparticles (CT-LNP) loading S100A4 siRNA (CT-LNP-siA4) to silence the S100A4 gene in OA chondrocytes. In both mouse and rat OA models, CT-LNP-siA4 could significantly downregulate the expression of S100A4 and OA phenotype-related molecules, such as COL I, MMP-13, and IL-6, inhibiting OA progression and chronic pain. This study validated S100A4 as a potential molecular target and proved that the corresponding LNP drug formulation was effective for the OA treatment at various animal models.

Engineering the functional peptide self-assembly has been proven effective for drug delivery, creating three-dimensional cell culture media and developing new strategies for disease therapy. However, there are few reports on using peptide assemblies as nanotechnological tools to explore the processes and mechanisms of biology. In this work, to investigate tumor lysosomal membrane permeabilization (LMP)-induced effect, which is considered as a promising but not well-defined strategy for treatment of cancers, we established a tumor-specific LMP model by rational construction of a pH-responsive morphology transformable self-assembly of amphiphilic oligopeptide (AOP), containing -[Arg-Gly-Asp]- (RGD) sequence. In brief, the selected AOP, Benz-(Ala)6-Arg-Gly-Asp-NH2 (Benz-A6-RGD), could self-assemble to liposome-like nanostructures (peptosomes) at neutral pH (7.4), and the RGD motifs on the surface of peptosomes could recognize integrins on tumor cells and enhance the following endocytosis; then the lysosomal pH (4.0-5.0) protonized RGD motifs and induced the peptosomes to transform to nanofibers. This transformation produced mechanical forces to directly disturb the membrane of lysosomes so as to initiate LMP. To further enhance the antitumor effect, the LMP-induced cell death was combined with the inhibition of the hot shock protein70 (Hsp70)-mediated self-repair mechanism of tumor cells. A significant synergetic antitumor effect was observed for this combination strategy. In summary, the current study introduces a specific model of tumor cell LMP, which can be used for evaluating the LMP-induced effects on tumor cells, and proves the potential of functional peptide self-assembly for exploring biological processes.

Neuropathic pain (NP) is a debilitating condition driven by chronic neuroinflammation, where abnormal communication between microglia and astrocytes amplifies pain signaling. Current therapies offer limited benefit and primarily address symptoms rather than underlying mechanisms. Here, we show that transferrin- and phosphatidylserine-modified liposomes carrying a TDP-43 aggregation inhibitor (TF/PS/TDP-43-IN-1) effectively cross the blood-brain barrier, target glial cells, and modulate their activation states. In vitro, the formulation improved cell viability and promoted anti-inflammatory phenotypes. In vivo, studies conducted in male C57BL/6 J mice demonstrated significant alleviation of pain behaviors, reduced inflammatory cytokine expression, and suppression of the cGAS-STING pathway. These findings indicate that targeting TDP-43 aggregation with a nanocarrier system can reprogram glial interactions to relieve neuropathic pain. This strategy highlights a promising approach for developing targeted, disease-modifying therapies that act on key drivers of neuroinflammation.

Lipid nanoparticles (LNPs) are the most clinically relevant vehicles for mRNA vaccines. Despite the great successes, the toxicity caused by the high dose of lipid components still represents a great challenge. The suboptimal loading capacity of mRNA in LNPs not only compromises the vaccine's efficacy but also heightens the risk of non-specific immune responses, accelerates clearance caused by anti-PEG IgG/IgM. These problems underscore the urgent need for improving mRNA loading capacity in LNPs to provide dose-sparing effects. Herein, we develop a metal ion mediated mRNA enrichment strategy to efficiently form a high-density mRNA core, and manganese ion (Mn2+) exhibits a unique capability to match the need. The prepared Mn-mRNA nanoparticle is subsequently coated with lipids to form the resulting nanosystem, L@Mn-mRNA, which achieved nearly twice the mRNA loading capacity compared to conventional mRNA vaccine formulations (LNP-mRNA). Remarkably, L@Mn-mRNA also demonstrates a 2-fold increase in cellular uptake efficiency compared to LNP-mRNA, attributed to the enhanced stiffness provided by the Mn-mRNA core. By combining improved mRNA loading with superior cellular uptake, L@Mn-mRNA achieves significantly enhanced antigen-specific immune responses and therapeutic efficacy as vaccines. We elucidate the mechanism behind Mn-mRNA construction and optimize the L@Mn-mRNA formulations, and this method is suitable for types of lipids and mRNAs. Moreover, L@Mn-mRNA also reduces the risk of anti-PEG IgG/IgM generation. Thus, this strategy holds significant potential as a platform for the next generation of lipid-based mRNA vaccines.

Catalytic therapy with enzymes and nanozymes is a promising approach to cancer treatment. However, it often proves inadequate when used as a standalone treatment. This work presents the catalase (CAT)-like activity of Ti3C2Tx MXene, a kind of two-dimensional (2D) material with photothermal capability. Ti3C2Tx nanosheets were then used to load glucose oxidase (GOx) and the anticancer drug doxorubicin hydrochloride (DOX), and modified with polyethylene glycol (PEG) to obtain an enzyme cascade system, named Ti3C2Tx-GOx/DOX-PEG, for anticancer purposes. Ti3C2Tx-GO(x)/DOX-PEG exhibited a photothermal conversion efficiency of similar to 28.7% at 808 nm, enabling its use for near-infrared light-based photothermal therapy (PTT). In addition, Ti3C2Tx could decompose H2O2 to O-2 in cancer cells, thereby enhancing starvation therapy by sensitizing GOx. Furthermore, alleviation of hypoxia could sensitize the chemotherapy of DOX. The decomposition of glucose by GOx could also generate H2O2 for the production of O-2 by Ti(3)C(2)Tx. Ultimately, the cycling of the enzyme cascade reaction could alleviate tumor hypoxia and enable enhanced chemo/starvation/photothermal combination therapy. The CAT-like behavior of Ti3C2Tx as well as various enzymatic reactions catalyzed by Ti3C2Tx-GOx/DOX-PEG were systematically investigated by intra- and extracellular experiments. Results from both in vitro and in vivo studies demonstrated the effective anticancer capability of Ti3C2Tx-GOx/DOX-PEG. This enzyme cascade system has the potential to enhance material utilization and reduce side effects, thus opening up new opportunities for designing efficient and safe anticancer systems.

Hand, foot, and mouth disease (HFMD), a common childhood infection caused by enterovirus, poses a serious public health concern in China. We collected and analyzed epidemiological data on 62,133 HFMD cases in Shenyang City, Liaoning Province, from 2013 to 2023. The average annual incidence was 76.12 per 100,000 person-years; 99.45% of cases were mild, while 0.55% were severe. Only one patient died. HFMD infections peaked annually in July. Children in kindergartens and scattered children accounted for 44.6% and 42.2% of cases, respectively. Real-time RT-PCR detection of enteroviruses in 5534 patient samples revealed 3780 positives, of which 25.1% were CVA16-positive. Positives were randomly sampled, yielding 240 VP1 sequences of CVA16. Phylogenetic tree results showed that all VP1 sequences belonged to the B1 sub-genogroup. However, the sub-genogroup prevalence varied over time: from 2013 to 2014 and 2019 to 2021, the predominant sub-genogroup was B1a, while it was B1b from 2015 to 2018. Further phylogenetic analyses showed substantial divergence between B1a branches in CVA16, suggesting possible turnover of the B1a sub-genogroup in CVA16 due to evolution. This study provides epidemiological data on HFMD in Shenyang, and provides a phylogenetic analysis of CVA16, offering a theoretical basis for preventing and controlling HFMD in Shenyang City.