Nano TodayPub Date : 2025-04-02DOI: 10.1016/j.nantod.2025.102737
Suleixin Yang , Ruie Chen , Peng Hua , Yi Wu , Meiwan Chen
{"title":"Integrating autophagy inhibition and ROS clearance in biohybrid nanoparticles for low-temperature cancer photothermal therapy","authors":"Suleixin Yang , Ruie Chen , Peng Hua , Yi Wu , Meiwan Chen","doi":"10.1016/j.nantod.2025.102737","DOIUrl":"10.1016/j.nantod.2025.102737","url":null,"abstract":"<div><div>Photothermal therapy (PTT) demands efficient cancer ablation at relative low temperatures and minimal thermal damage to normal tissues, but suffers from both the protective autophagy-related thermal resistance in cancer cell and reactive oxygen species (ROS)-induced damage to normal cells. Here, we screened out curcumin-Fe ultrasmall nanoparticles (Cur-Fe) that manifested efficient photothermal conversion efficiency (η = 43.38 %) and ROS scavenging ability. Additionally, CRISPR/Cas9 plasmids (pCas-ATG5/ATG7) were also constructed to safely and precisely knockdown the protective autophagy for thermal resistance alleviation. The core Cur-Fe/ATG@<sup>TK</sup>PF (CFA@T), which was composed of anionic <u>C</u>ur-<u>F</u>e and pCas-<u>A</u>TGs, was encapsulated by cationic thioketal-crosslinked and fluorinated polyethyleneimine (<sup><u>T</u>K</sup>PF) via electrostatic interaction. Further, CFA@TC was formed by CFA@T coated with an acidic pH-responsive shell OH<u>C</u>-PEG-CHO via Schiff base. Attributed to its dual responsiveness to pH and ROS, CFA@TC exhibited efficient tumor targeting and uptake following intravenous injection. Upon irradiation with a 652 nm laser, CFA@TC demonstrated enhanced efficacy in eradicating cancer cells by inhibiting autophagy, while concurrently mitigating inflammatory responses through intracellular ROS scavenging <em>in vivo</em> and <em>in vitro</em>. Taken together, our study provides a proof-of-concept that CRISPR can be effective for autophagy inhibition, and its integration with ROS-induced inflammatory responses relief can further improve PTT.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"63 ","pages":"Article 102737"},"PeriodicalIF":13.2,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143747398","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nano TodayPub Date : 2025-04-01DOI: 10.1016/j.nantod.2025.102731
Xiaoxue Xie , Jing Liu , Zhengjie Liu , Huiye Wei , Minzhao Lin , Gengjia Chen , Zhibo Liu , Mengyi He , Xinshuo Huang , Shuang Huang , Yunuo Wang , Ji Wang , Huijiuan Chen , Qi Chen , Xi Xie , Xintao Shuai
{"title":"Real-time ROS monitoring-guided tumor electrodynamic therapy using a metal microneedle array system","authors":"Xiaoxue Xie , Jing Liu , Zhengjie Liu , Huiye Wei , Minzhao Lin , Gengjia Chen , Zhibo Liu , Mengyi He , Xinshuo Huang , Shuang Huang , Yunuo Wang , Ji Wang , Huijiuan Chen , Qi Chen , Xi Xie , Xintao Shuai","doi":"10.1016/j.nantod.2025.102731","DOIUrl":"10.1016/j.nantod.2025.102731","url":null,"abstract":"<div><div>Currently, various strategies are employed to utilize reactive oxygen species (ROS) amplifiers in tumor therapy; Electrodynamic therapy (EDT) presents a promising modality for ROS amplification, as it can continuously produce substantial quantities of ROS independent of endogenous sources, thereby demonstrating potential antitumor activity. Nonetheless, achieving prolonged tumor suppression with EDT remains a significant challenge. Conventional EDT approaches frequently encounter issues with inadequate overlap between the active electric field region and the drug distribution area, resulting in insufficient electrocatalytic action and uneven ROS distribution. Furthermore, individual physiological variability can lead to disparate therapeutic outcomes from identical drug dosages, and indiscriminate increases in ROS dosage may inadvertently exacerbate tumor invasion and metastasis. To address these challenges, we developed a microneedle (MN) array system that combines ROS sensing and enables precise EDT therapy. The integrated system offers multifunctional capabilities, including drug delivery, electrical stimulation, and real-time ROS sensing. Benefiting from the homogeneously distributed electric field provided by the MN array, we significantly enhanced the electrocatalytic performance of electrodynamic nanomedicines. The integrated system produces cell-toxic ROS at 2.4 times the rate of traditional methods and induces tumor cell apoptosis 2.6 times more effectively. Real-time ROS monitoring via sensing electrodes allows precise drug dosage adjustments, ensuring effective ROS amplification therapy while minimizing waste. Adding DON further boosts ROS accumulation and strengthens anti-tumor immunity. The miniaturized dual-power supply strategy, combining a constant current source with ROS signal collection, enhances clinical suitability, optimizing both therapeutic efficacy and precision in tumor treatment.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"63 ","pages":"Article 102731"},"PeriodicalIF":13.2,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143747528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nano TodayPub Date : 2025-03-31DOI: 10.1016/j.nantod.2025.102728
Jianlin Ge , Hu Chen , Yao Zhang , Longyi Zheng , Yanyu Miao , Jinyang Wang , Wenhui Wu , Yulun Chen , Xiaoyu An , Xuqi Peng , Minglei Teng , Hui Liu , Jianzhong Zhang , Chao Liu , Ping Xu , Gang Liu
{"title":"BCG emulsified in lipiodol expands tumor epitope profile to boost cancer immunotherapy","authors":"Jianlin Ge , Hu Chen , Yao Zhang , Longyi Zheng , Yanyu Miao , Jinyang Wang , Wenhui Wu , Yulun Chen , Xiaoyu An , Xuqi Peng , Minglei Teng , Hui Liu , Jianzhong Zhang , Chao Liu , Ping Xu , Gang Liu","doi":"10.1016/j.nantod.2025.102728","DOIUrl":"10.1016/j.nantod.2025.102728","url":null,"abstract":"<div><div>The limited shared antigens identified in cancers are insufficient as targets for universal immunotherapy, and the impaired major histocompatibility complex class I (MHC-I) expression, limited antigen exposure, and deficiency of immunogenicity lead to tumors evading immune surveillance. Here, we proposed a facile and effective BCG emulsion immunotherapy strategy that expands antigen exposure, increasing immune recognition, and boosting cancer immunotherapy. BCG emulsion, exhibiting efficient internalization in a non-fibronectin-dependent manner, reduced tumor growth, reprogrammed tumor microenvironment, and enhanced signaling pathways related to antigen processing and presentation. BCG emulsion can reverse the MHC-I downregulation, and peptide fragments from BCG-derived molecules are presented on the tumor cell surface by MHC-I. CD8<sup>+</sup> T cells recognize and are activated against these presented BCG epitopes identified by immunopeptideome. The BCG dominant epitope emulsion induced significant infiltration of CD8<sup>+</sup>T cells and dendritic cells in tumors. Overall, this study demonstrated for the first time the potential of BCG and BCG epitope administration in anti-cancer applications and provided new technological methods and ideas for cancer immunotherapy.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"62 ","pages":"Article 102728"},"PeriodicalIF":13.2,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143747702","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nano TodayPub Date : 2025-03-30DOI: 10.1016/j.nantod.2025.102745
Ali Akbar Ashkarran
{"title":"Decentralized nanoparticle protein corona analysis may misconduct biomarker discovery","authors":"Ali Akbar Ashkarran","doi":"10.1016/j.nantod.2025.102745","DOIUrl":"10.1016/j.nantod.2025.102745","url":null,"abstract":"<div><div>Protein/biomolecular corona (PC) is a layer of biomolecules (mainly proteins) that forms around the nanoparticles (NPs) after exposure to biological environments (e.g., blood). The dynamic nature of the PC formation allows enrichment of specific proteins particularly low abundance proteins and enables capturing disease-specific proteins on the surface of the NPs for biomarker discovery. However, identification of crucial proteins with high diagnostic values heavily depends on liquid chromatography mass spectrometry (LC-MS/MS) approach due to its sensitivity and specificity. Despite the widespread use of MS, there exist potential pitfalls in NPs’ PC analysis that may lead to misconduct in biomarker discovery studies. This opinion considers these pitfalls, providing insights into the challenges and strategies to mitigate misconduct in biomarker discovery using mass spectrometry techniques.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"62 ","pages":"Article 102745"},"PeriodicalIF":13.2,"publicationDate":"2025-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734554","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nano TodayPub Date : 2025-03-29DOI: 10.1016/j.nantod.2025.102739
Xiaoru Zhang , Shuiling Wen , Qin Liu , Wenli Cai , Keke Ning , Han Liu , Ergang Liu , Yongzhuo Huang , Feng Zeng
{"title":"Multi-functional nanozyme-integrated astragalus polysaccharide hydrogel for targeted phased therapy in diabetic wound healing","authors":"Xiaoru Zhang , Shuiling Wen , Qin Liu , Wenli Cai , Keke Ning , Han Liu , Ergang Liu , Yongzhuo Huang , Feng Zeng","doi":"10.1016/j.nantod.2025.102739","DOIUrl":"10.1016/j.nantod.2025.102739","url":null,"abstract":"<div><div>Diabetic wounds (DW) are characterized by excessive oxidative stress, chronic inflammation, hypoxia, impaired angiogenesis, weakened antioxidant defenses, and disrupted collagen remodeling, all of which delay healing and compromise tissue integrity. To address these challenges, we developed a biodegradable multifunctional hydrogel dressing (Fe/Ce@APS Gel) comprised of astragalus polysaccharide (APS), polyvinyl alcohol (PVA), and borax, functionalized with multi-enzyme mimetic nanozyme iron-modified ceria nanoparticles (Fe/CeNP-PEG). This Fe/Ce@APS Gel demonstrates potent anti-inflammatory, antioxidant, oxygenation, and pro-angiogenic properties, supporting wound healing across all stages. In the initial bleeding phase, the dressing accelerates blood clotting, promoting rapid wound stabilization. During the inflammatory phage, Fe/CeNP-PEG and APS effectively reduces excess reactive oxygen species (ROS) generates oxygen, modulates macrophage polarization, and mitigates inflammatory responses. In the proliferative phase, APS enhances cell proliferation, stimulates angiogenesis, and accelerates granulation tissue formation, supporting tissue repair. Finally, in the remodeling phase, Fe/Ce@APS Gel aids in tissue architecture reconstruction, strengthening wound integrity. Mechanistically, Fe/Ce@APS Gel facilitates DW healing by inhibiting the NLRP3/NF-κB signaling pathway, thereby reducing inflammation. The synergistic effects of APS and Fe/CeNP-PEG underscore the potential of Fe/Ce@APS Gel as a promising therapeutic dressing for DW treatment.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"62 ","pages":"Article 102739"},"PeriodicalIF":13.2,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734555","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nano TodayPub Date : 2025-03-28DOI: 10.1016/j.nantod.2025.102742
Shengke Zhao , Yunjian Yu , Youtao Xin , Hegang Lu , Xiaohui Li , Shuyu Wang , Feihe Ma , Hui Gao
{"title":"Oral delivery of Clostridium butyricum using selective antibacterial lipids for enhanced treatment of Fusobacterium nucleatum-associated intestinal diseases","authors":"Shengke Zhao , Yunjian Yu , Youtao Xin , Hegang Lu , Xiaohui Li , Shuyu Wang , Feihe Ma , Hui Gao","doi":"10.1016/j.nantod.2025.102742","DOIUrl":"10.1016/j.nantod.2025.102742","url":null,"abstract":"<div><div>The gut microbiota plays a crucial role in host immune modulation and maintaining homeostasis. An abnormal increase in certain pathogens such as <em>Fusobacterium nucleatum</em> (<em>Fn</em>) can break homeostasis and drive the progression of various intestinal diseases. Supplementing probiotics can partially counteract these effects without flora disturbance. However, broad-spectrum antibacterial treatments are not compatible with probiotic therapy in a therapeutic system due to their non-selective damage on both probiotics and the overall gut microbiota. Herein, we screen and identify lauric acid (LA)-derived lipid, S12, from a combinatorial library of 12 chemically diverse lipids for its selective antibacterial activity against <em>Fn</em> over probiotic <em>Clostridium butyricum</em> (<em>Cb</em>). This lipid is then utilized as a single-cell carrier to orally deliver <em>Cb</em> (<em>Cb</em>@S12) for enhanced treatment of <em>Fn</em>-associated intestinal diseases. The surface arming of S12 effectively protects <em>Cb</em> from simulated gastric and intestinal fluids, thus significantly prolonging its intestinal retention in mice. Oral administration of <em>Cb</em>@S12 has demonstrated impressive therapeutic outcomes against <em>Fn</em>-aggravated inflammatory bowel disease and orthotopic colorectal cancer by selectively eliminating <em>Fn</em> while preserving the probiotic activity of <em>Cb</em>. This study introduces a robust approach using selectively antibacterial lipids for probiotic encapsulation, offering an antibiotic-free “probiotic-antagonistic” combination therapeutic strategy for intestinal diseases.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"62 ","pages":"Article 102742"},"PeriodicalIF":13.2,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143724654","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nano TodayPub Date : 2025-03-28DOI: 10.1016/j.nantod.2025.102730
Yang Cao , Yan Liu , Xiaoying Shang , Yao Lin , Lushan Lin , Ni Zhang , Hang Gao , Xueyuan Chen
{"title":"Full-color circularly polarized luminescence from perovskite quantum dots embedded within Chiral ZIF-8 matrix","authors":"Yang Cao , Yan Liu , Xiaoying Shang , Yao Lin , Lushan Lin , Ni Zhang , Hang Gao , Xueyuan Chen","doi":"10.1016/j.nantod.2025.102730","DOIUrl":"10.1016/j.nantod.2025.102730","url":null,"abstract":"<div><div>The exploration of circularly polarized luminescence (CPL) materials based on perovskite quantum dots (PeQDs) has garnered significant interest across various disciplines owing to their extensive potential in optical applications. However, conventional perovskite-based CPL materials frequently encounter formidable challenges, including complex fabrication processes, limited emission bandwidths, inevitable anion exchange, and aggregation-induced quenching. To address these challenges, we proposed a unique approach to develop solid-state CPL nanohybrids with superior full-color CPL by integrating CsPbX<sub>3</sub> (X = Cl, Br, I) PeQDs into amino acid co-assembled chiral metal-organic frameworks (CMOFs). By in situ generating PeQDs within L/D-ZIF-8 CMOFs, we achieved solid-state CPL nanohybrids (L/D-ZIF-8⊃PeQDs) that exhibited enhanced CPL properties and stability. The chiral microenvironment provided by the CMOFs not only boosts CPL performance but also effectively mitigates issues such as anion exchange and aggregation-induced quenching. More intriguingly, such nanohybrids displayed tunable CPL emissions across the entire visible spectrum, achieving a maximum dissymmetry factor (<em>g</em><sub>lum</sub>) value of 1.41 × 10<sup>−3</sup> and a photoluminescence quantum yield of up to 13 %. Furthermore, we showcased their proof-of-concept application by fabricating circularly polarized red-green-blue and white light-emitting diodes with an impressive color gamut exceeding 137 % NTSC, thereby unveiling the significance of our approach in promoting CPL functionalities of perovskite-based materials.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"62 ","pages":"Article 102730"},"PeriodicalIF":13.2,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143714656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Nanozyme engineered ROS-tolerant cysteine active-site for upstream deubiquitylation therapy of inflammatory bowel disease","authors":"Haibin Wu , Shuhan Shi , Lenan Xu , Ziying Zheng , Ziyan Huang , Yi Qiu , Jixiang Zhang , Keyun Yang , Yuting Xie , Cheng Xu , Tianyang Xu , Guohuan Zeng , Lingfeng Chen , Mincong Huang , Qian Chen , Daishun Ling , Guang Liang","doi":"10.1016/j.nantod.2025.102735","DOIUrl":"10.1016/j.nantod.2025.102735","url":null,"abstract":"<div><div>Deubiquitinase (DUB)-based upstream regulation of inflammatory pathway has emerged as a promising strategy for inflammatory bowel disease (IBD) compared to clinical biological therapies that block downstream inflammatory mediators. Unfortunately, excessive reactive oxygen species (ROS) in the inflammatory microenvironment usually induces irreversible oxidation of the cysteine active-site of therapeutic DUB, resulting in the rapid fading of deubiquitylation activity and inferior anti-inflammatory efficacy. Herein, a redox active nanozyme boosted deubiquitylation strategy based on the natural-artificial dual catalytic nanodrug (NADCN) is designed for protecting the vulnerable catalytic active center of anti-inflammatory DUB. Benefiting from the nanozyme-enabled protection of DUB’s cysteine active-site, the NADCN exhibits far superior anti-inflammatory activity to that of its counterparts based on the conventional delivery system. Moreover, the redox nanozyme in NADCN confers efficient modulation of the oxidative and hypoxia inflammatory microenvironment, further unleashing the potential of DUB for treatment of IBD. Therefore, the NADCN opens a new avenue to protect the cysteine active-site of therapeutic DUB, providing an attractive opportunity for developing ROS-tolerant deubiquitylation therapy.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"62 ","pages":"Article 102735"},"PeriodicalIF":13.2,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143714657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nano TodayPub Date : 2025-03-26DOI: 10.1016/j.nantod.2025.102734
Hong Wang , Xuehe Lu , Jing Fan , Changping Yang , Hanyin Zhu , Jianbing Liu , Baoquan Ding
{"title":"A carrier-free DNA nanoplatform for efficient three-in-one tumor therapy in vivo","authors":"Hong Wang , Xuehe Lu , Jing Fan , Changping Yang , Hanyin Zhu , Jianbing Liu , Baoquan Ding","doi":"10.1016/j.nantod.2025.102734","DOIUrl":"10.1016/j.nantod.2025.102734","url":null,"abstract":"<div><div>Drug delivery systems, based on chemical modification and controlled self-assembly of nucleic acid, have played an important role in the treatment of various diseases. Herein, we report a chemically conjugated DNA nanoplatform for efficient three-in-one tumor therapy <em>in vivo</em>. In our design, five copies of hydrophobic chemo-drug (camptothecin, CPT) are conjugated together by a branched organic molecule with an additional arm to introduce a DNA aptamer for targeting (Apt-5CPT). Meanwhile, a photosensitizer (HPPH, for photodynamic therapy) is efficiently organized at the terminal of an antisense oligonucleotide (AS, for gene therapy) to obtain another amphiphilic monomer (HPPH-AS). After co-assembly, a carrier-free DNA nanostructure, with the combination of chemotherapy, photodynamic therapy, and gene therapy, can be efficiently constructed for drug delivery. Under laser irradiation, the generated reactive oxygen species (ROS) can further facilitate their lysosomal escape to achieve subsequent glutathione (GSH)-based drug release for three-in-one tumor therapy <em>in vivo</em>. This rationally developed DNA nanoplatform-based drug delivery system presents a new avenue for the development of tumor therapy.</div></div>","PeriodicalId":395,"journal":{"name":"Nano Today","volume":"62 ","pages":"Article 102734"},"PeriodicalIF":13.2,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143714654","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}