A dual-reference study design for understanding and improving AAV genome size analysis.

Recombinant adeno-associated virus (rAAV) is the leading platform of gene delivery for its long-lasting gene transformation and low immunogenicity. Characterization of the integrity and purity of the rAAV genome is critical to ensure clinical potency and safety. However, current rAAV genome characterization methods that can provide size assessment are either time-consuming or not easily accessible to general labs. Additionally, there is a lack of right reference standard for analyzing long single-stranded DNA (ssDNA) fragments. Here, we have developed an ssDNA assay on a microfluidic capillary electrophoresis platform using ssDNA reference standard. This assay provides size calling for ssDNA fragment, a detection sensitivity at ∼89 pg/µL (3 × 1010  GC/mL AAV) for 5.1 kb ssDNA fragment, and a turnaround time at ∼100 s per sample with a high throughput sample analyzing capability. Moreover, we have observed that the annealing of AAV ssDNA subsequent to its release from the capsid might introduce an additional double-stranded DNA (dsDNA) peak. This phenomenon is dependent on the sample processing workflow. To avoid the risk of mischaracterization, we recommend the use of dual-reference standards in combination with other orthogonal methods to have a comprehensive understanding of the rAAV genome size and integrity.

Comprehensive Overview of Controlled Fabrication of Multifunctional Fluorescent Carbon Quantum Dots and Exploring Applications.

In recent years, carbon dots (CDs) have garnered increasing attention due to their simple preparation methods, versatile performances, and wide-ranging applications. CDs can manifest various optical, physical, and chemical properties including quantum yield (QY), emission wavelength (Em), solid-state fluorescence (SSF), room-temperature phosphorescence (RTP), material-specific responsivity, pH sensitivity, anti-oxidation and oxidation, and biocompatibility. These properties can be effectively regulated through precise control of the CD preparation process, rendering them suitable for diverse applications. However, the lack of consideration given to the precise control of each feature of CDs during the preparation process poses a challenge in obtaining the requisite features for various applications. This paper is to analyze existing research and present novel concepts and ideas for creating CDs with different distinct features and applications. The synthesis methods of CDs are discussed in the first section, followed by a comprehensive overview of the important properties of CDs and the modification strategy. Subsequently, the application of CDs and their requisite properties are reviewed. Finally, the paper outlines the current challenges in controlling CDs properties and their applications, discusses potential solutions, and offers suggestions for future research.

Bioinspired Tumor Calcification-Guided Early Diagnosis and Eradication of Hepatocellular Carcinoma.

Tumor calcification is found to be associated with the benign prognostic, and which shows considerable promise as a somewhat predictive index of the tumor response clinically. However, calcification is still a missing area in clinical cancer treatment. A specific strategy is proposed for inducing tumor calcification through the synergy of calcium peroxide (CaO2)-based microspheres and transcatheter arterial embolization for the treatment of hepatocellular carcinoma (HCC). The persistent calcium stress in situ specifically leads to powerful tumor calcioptosis, resulting in diffuse calcification and a high-density shadow on computed tomography that enables clear localization of the in vivo tumor site and partial delineation of tumor margins in an orthotopic HCC rabbit model. This osmotic calcification can facilitate tumor clinical diagnosis, which is of great significance in differentiating tumor response during early follow-up periods. Proteome and phosphoproteome analysis identify that calreticulin (CALR) is a crucial target protein involved in tumor calcioptosis. Further fluorescence molecular imaging analysis also indicates that CALR can be used as a prodromal marker of calcification to predict tumor response at an earlier stage in different preclinical rodent models. These findings suggest that upregulated CALR in association with tumor calcification, which may be broadly useful for quick visualization of tumor response.

Internal Light Sources-Mediated Photodynamic Therapy Nanoplatforms: Hope for the Resolution of the Traditional Penetration Problem.

Photodynamic therapy (PDT) is an alternative cancer treatment technique with a noninvasive nature, high selectivity, and minimal adverse effects. The indispensable light source used in PDT is a critical factor in determining the energy conversion of photosensitizers (PSs). Traditional light sources are primarily concentrated in the visible light region, severely limiting their penetration depth and making them prone to scattering and absorption when applied to biological tissues. For that reason, its efficacy in treating deep-seated lesions is often inadequate. Self-exciting PDT, also known as auto-PDT (APDT), is an attractive option for circumventing the limited penetration depth of traditional PDT and has acquired significant attention. APDT employs depth-independent internal light sources to excite PSs through resonance or radiative energy transfer. APDT has considerable potential for treating deep-tissue malignancies. To facilitate many researchers' comprehension of the latest research progress in this field and inspire the emergence of more novel research results. This review introduces internal light generation mechanisms and characteristics and provides an overview of current research progress based on the recently reported APDT nanoplatforms. The current challenges and possible solutions of APDT nanoplatforms are also presented and provide insights for future research in the final section of this article.

A silver metal-organic cage with antibacterial activity for wound healing.

Bacterial infection is one of the most threatening diseases in humans and can result in tissue necrosis, inflammation, and so on. Although a large number of antibacterial materials have been developed, there are still some disadvantages in this field, including decreasing antibacterial activity in the aqueous solution or a short duration of time. Herein, a metal-organic cage named Ag-TBI-TPE with excellent antibacterial activity was prepared and applied in wound healing. Owing to the photosensitive production of the toxic ROS species and the positive charge of the surface, the Ag-TBI-TPE cage exhibits high antibacterial activity, especially under UV irradiation. It could accelerate the healing process of the infected wounds in vivo with satisfactory biocompatibility and bio-safety. The results indicated that after treatment with the Ag-TBI-TPE cage, with and without UV irradiation, the healing rates of wounds infected by E. coli and S. aureus were 89.59% and 93.05%, and 83.48% and 90.84%, respectively, which were much higher than those shown by the positive control group at 51.38% and 67.74%, respectively. This study not only sheds light on a design idea for a new antibacterial material but also further expands the potential application field of metal-organic cages.

Molecular Engineering of NIR-II/IIb Emitting AIEgen for Multimodal Imaging-Guided Photo-Immunotherapy.

In view of the great challenges related to the complexity and heterogeneity of tumors, efficient combination therapy is an ideal strategy for eliminating primary tumors and inhibiting distant tumors. A novel aggregation-induced emission (AIE) phototherapeutic agent called T-TBBTD is developed, which features a donor-acceptor-donor (D-A-D) structure, enhanced twisted molecule conformation, and prolonged second near-infrared window (NIR-II) emission. The multimodal imaging function of the molecule has significance for its treatment time window and excellent photothermal/photodynamic performance for multimode therapy. The precise molecular structure and versatility provide prospects for molecular therapy for anti-tumor applications. Fluorescence imaging in the NIR-II window offers advantages with enhanced spatial resolution, temporal resolution, and penetration depth. The prepared AIE@R837 NPs also have controllable performance for antitumor photo-immunotherapy. Following local photo-irradiation, AIE@R837 NPs generate abundant heat, and 1 O2 directly kills tumor cells, induces immunogenic cell death (ICD) as a photo-therapeutic effect, and releases R837, which enhances the synergistic effect of antigen presentation and contributes to the long-lasting protective antitumor immunity. A bilateral 4T1 tumor model revealed that this photo-immunotherapy can eliminate primary tumors. More importantly, it has a significant inhibitory effect on distant tumor growth. Therefore, this method can provide a new strategy for tumor therapy.

Integrated Nanorod-Mediated PD-L1 Downregulation in Combination with Oxidative-Stress Immunogene Therapy against Cancer.

It is an engaging program for tumor treatment that rationalizes the specific microenvironments, activation of suppressed immune system (immune resistance/escape reversion), and synergistic target therapy. Herein, a biomimetic nanoplatform that combines oxidative stress with genetic immunotherapy to strengthen the therapeutic efficacy is developed. Ru-TePt nanorods, small interfering RNA (PD-L1 siRNA), and biomimetic cellular membrane vesicles with the targeting ability to design a multifunctional Ru-TePt@siRNA-MVs system are rationally integrated. Notably, the Fenton-like activity significantly enhances Ru-TePt nanorods sonosensitization, thus provoking stronger oxidative stress to kill cells directly. Meanwhile, immunogenic cell death is triggered to secrete numerous cytokines and activate T cells. The effective catalase characteristics of Ru-TePt enable the in situ oxygen-producing pump to improve tumor oxygen level and coordinately strengthen the therapeutic effect of SDT followed. More importantly, anti-PD-L1-siRNA mediated immune checkpoint silence of the PD-L1 gene creates an environment conducive to activating cytotoxic T lymphocytes, synergistic with boosted reactive oxygen species-triggered antitumor immune response. The experimental results in vitro and in vivo reveal that the Ru-TePt@siRNA-MVs nanosystems can effectively activate the oxidative stress-triggered immune response and inhibit PD-1/PD-L1 axis-mediated immune resistance. Consequently, this orchestrated treatment paradigm provides valuable insights for developing potential oxidative stress and genetic immunotherapy.

Application of microfluidic chip electrophoresis for high-throughput nucleic acid fluorescence fragment analysis assays.

Nucleic acid fragment analysis via separation and detection are routine operations in molecular biology. However, analysis of small single-stranded nucleic acid fragments (<100nt) is challenging and mainly limited to labor-intensive polyacrylamide gel electrophoresis or high-cost capillary electrophoresis methods. Here we report an alternative method, a microfluidic chip electrophoresis system that provides a size resolution of 5nt and a detection time of one minute per sample of fluorescence-labeled DNA/RNA fragments. The feasibility of this system was evaluated by quantifying CRISPR-Cas9 cleavage efficiency and the detection resolution was evaluated by analyzing ssDNA/RNA adenylation and phosphorylation. We employed this system to study the RNA capping efficiency and double-stranded DNA unwinding efficiency in isothermal amplification as two examples for assay design and evaluation. The microfluidic chip electrophoresis system provides a rapid, sensitive, and high-throughput fluorescence fragment analysis (FFA), and can be applied for enzyme characterization, reaction optimization, and product quality control in various molecular biology processes.

Investigating the influences of concave depths on stormwater runoff and pollution retention of urban grasslands.

In this study, scale-based runoff plots of concave grasslands were designed and simulated rainfall experiments were conducted to investigate their retention effectiveness for runoff volume and pollutant loads, and to analyze the influences of concave depths on runoff and pollution retention of grasslands. Results showed that mean time to runoff of concave grasslands was 88.5 minutes, which was 5.3 times than that of flat grassland. Average peak flow rate of concave grasslands was reduced by 36.2% compared with flat grassland. Concaved grasslands averagely retained 58.2% of stormwater runoff. Deeper concave depths significantly increased runoff detention and retention performance of grasslands. Total suspended solids (TSS) load reduction rates of concave grasslands were ranged from 50.8% to 97.3%. Total nitrogen (TN) load reduction rate was 49.8% for concave depth of 10 cm. Total phosphorus (TP) load reduction rates were 45.0% and 93.9% for grasslands with 5 cm and 10 cm concave depths, respectively. Pollution load reduction rates of TSS, TN and TP enhanced along with the increase in concave depths. The estimated minimum area ratios of upslope impervious surface to grasslands of 5 cm and 10 cm concave depths were approximately 1:1 under 20 mm rainfall events, and 38:1 under 5 mm rainfalls, respectively.

Role of the Global Fitness Regulator Genes on the Osmotic Tolerance Ability and Salinity Hazard Alleviation of Trichoderma asperellum GDFS 1009 for Sustainable Agriculture.

Velvet family proteins are global regulators of fungal growth and development. Here, we reported the role of Vel1 and Lae1 from T. asperellum in osmotic tolerance. Deletion of the Vel1 and Lae1 genes led to the retardation of vegetative mycelial growth under saline conditions. The strain carrying the overexpression locus of the Vel1 and Lae1 genes was highly resistant to oxidative stress by upregulating the enzymes and genes involved in antioxidant activity. Major physiological changes in the cell wall and vacuoles occurred under high saline conditions. The Vel1 and Lae1 overexpression strains increased cell wall thickness and the number of vacuoles, which seems to lead to an increase of the osmolyte content of glycerol and proline. The absorption of Na+ content in the vacuole of the Vel1 and Lae1 overexpression strains was increased, while the absorption of Na+ was impaired in the Vel1 and Lae1 knock out strains, in which the Na+ was localized in the cell wall membrane. This result supported the significant correlation of the expression of genes with the ionic transportation in T. asperellum. Maize root colonization by the Vel1 and Lae1 gene overexpression strain was increased, which would mitigate the stress caused by the absorption of Na+ in the maize roots and increased the plant growth. Our results highlighted the importance of Vel1 and Lae1 proteins to the salinity stress tolerance of T. asperellum and the mitigation of Na+ stress to plants for sustainable agriculture.

Broadening the Horizons of RNA Delivery Strategies in Cancer Therapy.

RNA-based therapy is a promising and innovative strategy for cancer treatment. However, poor stability, immunogenicity, low cellular uptake rate, and difficulty in endosomal escape are considered the major obstacles in the cancer therapy process, severely limiting the development of clinical translation and application. For efficient and safe transport of RNA into cancer cells, it usually needs to be packaged in appropriate carriers so that it can be taken up by the target cells and then be released to the specific location to perform its function. In this review, we will focus on up-to-date insights of the RNA-based delivery carrier and comprehensively describe its application in cancer therapy. We briefly discuss delivery obstacles in RNA-mediated cancer therapy and summarize the advantages and disadvantages of different carriers (cationic polymers, inorganic nanoparticles, lipids, etc.). In addition, we further summarize and discuss the current RNA therapeutic strategies approved for clinical use. A comprehensive overview of various carriers and emerging delivery strategies for RNA delivery, as well as the current status of clinical applications and practice of RNA medicines are classified and integrated to inspire fresh ideas and breakthroughs.

Basin management inspiration from impacts of alternating dry and wet conditions on water production and carbon uptake in Murray-Darling Basin.

The impacts of alternating dry and wet conditions on water production and carbon uptake at different scales remain unclear, which limits the integrated management of water and carbon. We quantified the response of runoff efficiency (RE) and plant water-use efficiency (PWUE) to a typical shift from dry to wet episode of 2003-2014 in Australia's Murray-Darling basin using good and specific data products for local application, including Australian Water Availability Project, Penman-Monteith-Leuning Evapotranspiration V2 product, MODIS MCD12Q1 V6 Land Cover Type and MODIS MOD17A3 V055 GPP product. The results show that there are significant power function relationships between RE and precipitation for basin and all ecosystems, while the PWUE had a negative quadratic correlation with precipitation and satisfied the significance levels of 0.05 for basin and the ecosystems except the grassland and cropland. The shrubs can achieve the best water production and carbon uptake under dry conditions, while the evergreen broadleaf trees and evergreen needleleaf trees can obtain the best water production and carbon uptake in wet conditions, respectively. These findings help integrated basin management for balancing water resource production and climate change mitigation.

Amplifying Dendritic Cell Activation by Bioinspired Nanometal Organic Frameworks for Synergistic Sonoimmunotherapy.

Despite recent advancements of sonodynamic therapy (SDT) in cancer immunotherapy, challenges have yet to be surmounted to further boost its immunotherapeutic efficacy due to the low-level tumor antigens presentation of dendritic cells (DCs). Cell membrane camouflaged-nanoparticles can integrate the neoantigens of the cancer cell membrane with the multifunctionalities of synthetic nanocores. Herein, sono-responsive nanoparticles coated with DC-targeted antibody chimeric cancer cell membrane are investigated for multimodal therapy. The nanometal organic frameworks (MOFs) that respond to ultrasound are loaded successfully inside the vesicles displaying an anti-DEC205 antibody. The anti-DEC205 chimeric vesicles can directly target and activate DCs, promote tumor antigens cross-presentation, and then produce a cascade amplified T-cell immune response. Upon deep tissue-penetrating sonication, AMR-MOF@AuPt generates large amounts of reactive oxygen species that directly kill cancer cells, further initiating an anti-cancer T cell immune response. Such synergistic sono-immunotherapies effectually inhibit tumor growth and induce strong systemic and long-term immune memory against cancer recurrence and distant metastasis. The authors findings provide DCs and tumor cells of a dual active-targeting cell membrane-coated sono-immunotherapeutic nanoplatform for cancer therapy.

Connecting Calcium-Based Nanomaterials and Cancer: From Diagnosis to Therapy.

As the indispensable second cellular messenger, calcium signaling is involved in the regulation of almost all physiological processes by activating specific target proteins. The importance of calcium ions (Ca2+) makes its "Janus nature" strictly regulated by its concentration. Abnormal regulation of calcium signals may cause some diseases; however, artificial regulation of calcium homeostasis in local lesions may also play a therapeutic role. "Calcium overload," for example, is characterized by excessive enrichment of intracellular Ca2+, which irreversibly switches calcium signaling from "positive regulation" to "reverse destruction," leading to cell death. However, this undesirable death could be defined as "calcicoptosis" to offer a novel approach for cancer treatment. Indeed, Ca2+ is involved in various cancer diagnostic and therapeutic events, including calcium overload-induced calcium homeostasis disorder, calcium channels dysregulation, mitochondrial dysfunction, calcium-associated immunoregulation, cell/vascular/tumor calcification, and calcification-mediated CT imaging. In parallel, the development of multifunctional calcium-based nanomaterials (e.g., calcium phosphate, calcium carbonate, calcium peroxide, and hydroxyapatite) is becoming abundantly available. This review will highlight the latest insights of the calcium-based nanomaterials, explain their application, and provide novel perspective. Identifying and characterizing new patterns of calcium-dependent signaling and exploiting the disease element linkage offer additional translational opportunities for cancer theranostics.

The Interpretation of the Role of a Polyketide Synthase ClPKS18 in the Pathogenicity of Curvularia lunata.

Plant pathogenic fungus Curvularia lunata (Wakker) Boedijn causes leaf spot diseases in several plants such as Oryza sativa, Sorghum bicolor (L.) Moench, and Capsicum frutescens. It has been spread worldwide, specifically in maize-growing regions. The polyketide synthase (PKS) plays a significant role in secondary metabolite production and its effect on virulence. The Clpks18 of C. lunata strongly correlated with its pathogenicity. The role of Clpks18 gene on the pathogenic activity of C. lunata remains unclear. Hence, in this study, we analyzed the importance of Clpks18 gene on the hyphae and conidial melanization and on the sporulation and hyphal growth. The deletion of Clpks18 gene reduced the production of methyl 5-(hydroxymethyl)furan-2-carboxylate toxin. The virulence of ΔClpks18 mutant was significantly reduced compared with the wild type. The metabolomics data revealed that (R)-(-)-mellein was a vital factor in the virulence of C. lunata. The (R)-(-)-mellein and the toxin produced by C. lunata were detected in the maize leaves during its infestation. In addition, the metabolomic analysis showed that the Clpks18 gene influences glycerolipid, non-ribosomal peptide biosynthesis, and its metabolism. This study demonstrates that the Clpks18 gene is important for the pathogenicity of C. lunata by influencing the complex metabolic network.

Recent Advancements in Poor Graft Function Following Hematopoietic Stem Cell Transplantation.

Poor graft function (PGF) is a life-threatening complication that occurs after transplantation and has a poor prognosis. With the rapid development of haploidentical hematopoietic stem cell transplantation, the pathogenesis of PGF has become an important issue. Studies of the pathogenesis of PGF have resulted in some success in CD34+-selected stem cell boosting. Mesenchymal stem cells, N-acetyl-l-cysteine, and eltrombopag have also been investigated as therapeutic strategies for PGF. However, predicting and preventing PGF remains challenging. Here, we propose that the seed, soil, and insect theories of aplastic anemia also apply to PGF; CD34+ cells are compared to seeds; the bone marrow microenvironment to soil; and virus infection, iron overload, and donor-specific anti-human leukocyte antigen antibodies to insects. From this perspective, we summarize the available information on the common risk factors of PGF, focusing on its potential mechanism. In addition, the safety and efficacy of new strategies for treating PGF are discussed to provide a foundation for preventing and treating this complex clinical problem.

Endoscopic modified total laminoplasty for symptomatic lumbar spinal stenosis.

Context/objective: At present, there is no consensus on the most effective surgical method for treating symptomatic lumbar spinal stenosis (LSS). Total laminectomy, which is frequently used at this time, destroys the posterior midline structure, causing many postoperative complications. We have designed a new surgical approach instead of total laminectomy. In this paper, we aimed to describe the surgical method of endoscopic modified total laminectomy for lumbar spinal stenosis as well as to explore its early efficacy.Participants: Patients with symptomatic LSS who underwent endoscopic modified total laminoplasty between August 2016 and August 2017 were eligible for our study.Outcome measures: Before surgery and one year after surgery, we measured lower limb pain and back pain by visual analog scale (VAS), disability via Oswestry Disability Index (ODI), and severity of back pain according to the Japanese Orthopedic Association Score for Back Pain (JOA), while any complications were also assessed.Results: Endoscopic modified total laminoplasty was performed on 22 LSS patients, including eight males and 14 females(mean age = 59.3 ± 9.6 years). We found statistically significant differences before and one year after surgery for VAS lower limb pain and back pain, ODI and JOA scores(P < 0.001). Complications included intraoperative dural tears(n = 1),and weak fusion between the lamina and the vertebral body (n = 1).Conclusion: Endoscopic modified total laminectomy is a promising surgical approach which reduces patient suffering and improves patient quality of life.

Curvularia lunata and Curvularia Leaf Spot of Maize in China.

Curvularia leaf spot (CLS), primarily caused by Curvularia lunata (Wakker) Boedijn (C. lunata), is widely distributed in maize production regions in China. It occurs in all the developmental stages of maize and causes economic losses. The epidemic and yield loss estimation models have been constructed for the disease. C. lunata has obvious virulence differentiation and produces multiple virulence factors. CLS is managed by application of chemical and biological agents and by quantitative resistance conferred by 5 to 6 quantitative trait loci (QTL). This review summarizes research on the understanding of CLS biological characteristics, virulence factors of C. lunata, host resistance genetics, and disease management strategies in China.

Nanotransferrin-Based Programmable Catalysis Mediates Three-Pronged Induction of Oxidative Stress to Enhance Cancer Immunotherapy.

Current oxidative stress amplifying strategies for immunogenic cell death (ICD) promotion are mainly restricted to immune tolerance induced by adaptive cellular antioxidation, limited tumor-selectivity, and tumoral immunosuppression. Herein, a facile and efficient scenario of genetically engineering transferrin-expressing cell membrane nanovesicle encapsulated IR820-dihydroartemisinin nanomedicine (Tf@IR820-DHA) was developed to boost a-PD-L1-mediated immune checkpoint blocking (ICB) via synergetic triple stimuli-activated oxidative stress-associated ICD. We demonstrate that the engineered transferrin of Tf@IR820-DHA has excellent tumor targeting and Fe(III)-loading properties and thus delivered Fe(III) and IR820-DHA nanoparticles (NPs) to the lesion location effectively. We found that the self-carrying Fe(III)-mediated programmable catalysis of DHA and glutathione (GSH) depletion generated plenty of reactive oxygen species (ROS). Moreover, DHA also acted as an immunomodulator to decrease the number of T regulatory cells, thereby remodeling the tumor immune microenvironment and achieving double T cell activation. Furthermore, the IR820 molecule served as a competent sonosensitizer to produce ROS under ultrasound activation and guide precise immunotherapy via fluorescent/photoacoustic (FL/PA) imaging. Through its three-pronged delivery of stimuli-activated oxidative stress (DHA-induced chemodynamic therapy, catalysis-conferred GSH depletion, and IR820-mediated sonodynamic therapy), Tf@IR820-DHA caused high levels of targeted ICD. This significantly increased the proportions of IFN-γ-secreting T cells (CD4+ T and CD8+ T) and enhanced a-PD-L1-mediated ICB against primary and distant tumors, which represents a promising approach for cancer nanoimmunotherapy.

Biomimetic nanoparticles blocking autophagy for enhanced chemotherapy and metastasis inhibition via reversing focal adhesion disassembly.

BACKGROUND: Autophagy is a conserved catabolic process, which plays an important role in regulating tumor cell motility and degrading protein aggregates. Chemotherapy-induced autophagy may lead to tumor distant metastasis and even chemo-insensitivity in the therapy of hepatocellular carcinoma (HCC). Therefore, a vast majority of HCC cases do not produce a significant response to monotherapy with autophagy inhibitors. RESULTS: In this work, we developed a biomimetic nanoformulation (TH-NP) co-encapsulating Oxaliplatin (OXA)/hydroxychloroquine (HCQ, an autophagy inhibitor) to execute targeted autophagy inhibition, reduce tumor cell migration and invasion in vitro and attenuate metastasis in vivo. The tumor cell-specific ligand TRAIL was bioengineered to be stably expressed on HUVECs and the resultant membrane vesicles were wrapped on OXA/HCQ-loaded PLGA nanocores. Especially, TH-NPs could significantly improve OXA and HCQ effective concentration by approximately 21 and 13 times in tumor tissues compared to the free mixture of HCQ/OXA. Moreover, the tumor-targeting TH-NPs released HCQ alkalized the acidic lysosomes and inhibited the fusion of autophagosomes and lysosomes, leading to effective blockade of autophagic flux. In short, the system largely improved chemotherapeutic performance of OXA on subcutaneous and orthotopic HCC mice models. Importantly, TH-NPs also exhibited the most effective inhibition of tumor metastasis in orthotopic HCCLM3 models, and in the HepG2, Huh-7 or HCCLM3 metastatic mice models. Finally, we illustrated the enhanced metastasis inhibition was attributed to the blockade or reverse of the autophagy-mediated degradation of focal adhesions (FAs) including E-cadherin and paxillin. CONCLUSIONS: TH-NPs can perform an enhanced chemotherapy and antimetastatic effect, and may represent a promising strategy for HCC therapy in clinics.