Journal of biomedical nanotechnology最新文献_第10页

Cardiomyocyte Exosomal Nanovesicles Mediating p53 to Regulate Autophagy and Protect Myocardial Infarction 心肌细胞外泌体纳米囊泡介导p53调节自噬保护心肌梗死

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3646

M. Fan, Fei Xie, Jianfeng Li, Junquan Li, Ting Liang

{"title":"Cardiomyocyte Exosomal Nanovesicles Mediating p53 to Regulate Autophagy and Protect Myocardial Infarction","authors":"M. Fan, Fei Xie, Jianfeng Li, Junquan Li, Ting Liang","doi":"10.1166/jbn.2023.3646","DOIUrl":"https://doi.org/10.1166/jbn.2023.3646","url":null,"abstract":"This study explores the protective mechanism of cardiomyocyte exosome nanovesicles on myocardial infarction. Enzyme activity was measured and apoptosis of rat cardiomyocytes was assessed by TdT-mediated dUTP Nick-End Labeling (TUNEL) along with analysis of myocardial function by color\u0000 Doppler echocardiogram. In addition, triphenyl tetrazolium chloride (TTC) staining evaluated the myocardial ischemia. BMSCs were co-cultured with primary cardiomyocytes followed by measuring autophagy and exosome morphology of myocardial tissue by transmission electron microscope and protein\u0000 expression by Western blot. Exosomes secreted by Bone marrow mesenchymal stem cells (BMSCs) were successfully isolated. LDH, CK, CK-MB activity, cardiomyocyte apoptosis rate, LVESD, LVEDD, LVEDP, myocardial infarction area, cell area occupied by autophagic vesicles and p53 expression in model\u0000 group were higher than control group (P <0.05) and they were lower in exosome group than model group. In addition, LVEF, LVFS, and LVSP were lower in model group and higher in exosome group (P <0.05). Model group showed significantly promoted autophagy activation and autophagosome\u0000 formation, which were inhibited by cardiomyocyte exosomes. Moreover, exosome-like vesicles were found with a diameter of 60–90 nm. Furthermore, PKH-26 staining showed that cardiomyocytes could uptake exosomes well. Myocardial injury significantly promotes autophagy activation. Moreover,\u0000 the apoptosis rate of cardiomyocytes and the area of myocardial ischemia were reduced and the cardiac function of rats with myocardial infarction was improved.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45138822","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Orthokeratology and Visual Motor Training for Juvenile Myopia 角膜塑形术与青少年近视的视觉运动训练

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3651

Yao Chen, Jingjing Zhang, Hui Huang, Bo Su

{"title":"Orthokeratology and Visual Motor Training for Juvenile Myopia","authors":"Yao Chen, Jingjing Zhang, Hui Huang, Bo Su","doi":"10.1166/jbn.2023.3651","DOIUrl":"https://doi.org/10.1166/jbn.2023.3651","url":null,"abstract":"This study aimed to explore the effect of orthokeratology (Ortho-K) combined with visual motor training in juvenile myopia. We included 156 myopia patients, which were randomly divided into the combined (visual motor training+Ortho-K) and Ortho-K (only) groups. Corneal morphological\u0000 changes (horizontal keratometry [HK], vertical keratometry [VK], central corneal thickness [CCT], and corneal endothelial cell density), regulatory function (positive and negative relative accommodation [PRA, NRA], regulatory amplitude, and regulatory sensitivity), ocular axis, visual acuity\u0000 (naked visual acuity, Diopter), tear film stability (BUT test and SIT test), intraocular pressure, and safety before and after treatment were compared between groups. Moreover, we performed a Spearman correlation analysis on the effects of initial age and diopter on axial growth. After treatment,\u0000 HK, VK of cornea, and CCT levels significantly decreased in both groups while no significant changes were found in endothelial cell density. In the combined group, there was a higher improvement in PRA, accommodation amplitude, and accommodation sensitivity, which could delay ocular axis growth\u0000 and improve naked eye vision and diopters. Meanwhile, a significant negative correlation was found between patients’ initial age and axial growth. Moreover, BUT and SIT recovery in the combined group were relatively better after wearing Ortho-K. In conclusion, Ortho-K combined with visual\u0000 motor training could better correct the vision of juvenile myopia patients and delay ocular axis growth. In addition, the earlier the intervention, the better the effect.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42654746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

NOX4 Upregulation in Lung Cancer: A Potential Therapeutic Target Associated with Immune Infiltration NOX4在肺癌中的上调:与免疫浸润相关的潜在治疗靶点

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3644

Hong Li, Sifang Feng, Tian Yang, Jiejuan Ning, Yuegang Wu, Tian-jun Chen

{"title":"NOX4 Upregulation in Lung Cancer: A Potential Therapeutic Target Associated with Immune Infiltration","authors":"Hong Li, Sifang Feng, Tian Yang, Jiejuan Ning, Yuegang Wu, Tian-jun Chen","doi":"10.1166/jbn.2023.3644","DOIUrl":"https://doi.org/10.1166/jbn.2023.3644","url":null,"abstract":"Lung cancer is a common and highly lethal tumor worldwide. Research indicates that NOX4 plays a crucial role in apoptosis resistance and sustained proliferation of cancer cells in various types of tumors. In this study, NOX4 expression in lung cancer tissues was analyzed using the Wilcoxon\u0000 rank sum test, while the Kruskal-Wallis Test was employed to explore the relationship between NOX4 expression and clinical characteristics in lung cancer patients. Prognostic evaluation was conducted using Kaplan-Meier plotter analysis, Cox regression, and receiver operating characteristic\u0000 (ROC) curve construction. Gene set enrichment analysis (GSEA) was performed to investigate the correlation between NOX4 and immune infiltration. Results showed significantly higher NOX4 expression in lung cancer patients compared to normal tissues. High NOX4 expression was associated with\u0000 shorter overall survival (OS) in lung cancer patients, as confirmed by Cox analysis. Furthermore, other clinicopathological factors predicted poor prognosis in lung adenocarcinoma (LUAD), and NOX4 demonstrated diagnostic value according to ROC analysis. Additionally, NOX4 overexpression correlated\u0000 with macrophages and Th1 cells based on SsGSEA analysis. In summary, NOX4 serves as an independent prognostic biomarker and is associated with immune infiltration in lung cancer.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41746015","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Insulin-Like Growth Factor 2 mRNA Binding Protein 3 Suppresses Ferroptosis in Non-Small Cell Lung Cancer via Stabilizing m6A Modification of Fanconi Anemia Group D2 Protein 胰岛素样生长因子2 mRNA结合蛋白3通过稳定范科尼贫血D2组蛋白的m6A修饰抑制非小细胞肺癌癌症中的铁蛋白脱失

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3643

Haotian Zhang, Junjie Tang, Xiaowei Gong, Chenjun Huang

{"title":"Insulin-Like Growth Factor 2 mRNA Binding Protein 3 Suppresses Ferroptosis in Non-Small Cell Lung Cancer via Stabilizing m6A Modification of Fanconi Anemia Group D2 Protein","authors":"Haotian Zhang, Junjie Tang, Xiaowei Gong, Chenjun Huang","doi":"10.1166/jbn.2023.3643","DOIUrl":"https://doi.org/10.1166/jbn.2023.3643","url":null,"abstract":"This study investigated the role of insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) in non-small cell lung cancer (NSCLC) and its association with N6-methyladenosine (m6A) modification. The study analyzes the expression levels and stability of IGF2BP3, as well as its impact\u0000 on NSCLC cell functions. The findings indicate that IGF2BP3 is upregulated in NSCLC patients and cell lines. Knocking down IGF2BP3 reduces cell proliferation and promotes ferroptosis in A549 and H1299 cells. Additionally, the study reveals that IGF2BP3 regulates the m6A modification of the\u0000 fanconi anemia group D2 protein (FANCD2) and influences its mRNA stability. Overexpressing FANCD2 counteracts the effects of IGF2BP3 silencing and increases the aggressiveness of NSCLC. Furthermore, treatment with celastrol induces ferroptosis in NSCLC cells and inhibits tumor growth in\u0000 vivo. In conclusion, these findings suggest that IGF2BP3 acts as an oncogene in NSCLC. Its interaction with FANCD2 through m6A modification suppresses ferroptosis in NSCLC cells. Thus, the IGF2BP3/FANCD2 signaling pathway may serve as a potential therapeutic target for NSCLC.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49522088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A Novel Silica Hybrid Nanoparticle with Zwitterion-Functionalized Polymer Chains for Highly Efficient N-Glycan Enrichment 一种具有两性离子功能化聚合物链的新型二氧化硅杂化纳米颗粒用于n-聚糖的高效富集

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3553

Yiting Pan, H. Bai, Guocheng Zhang, Wei Liu, Yanqi Wu, Hui Chen, Ying Tian, Zhenqi Yang, Duan Feng, Jiaqi Liu, Dan Wu

{"title":"A Novel Silica Hybrid Nanoparticle with Zwitterion-Functionalized Polymer Chains for Highly Efficient N-Glycan Enrichment","authors":"Yiting Pan, H. Bai, Guocheng Zhang, Wei Liu, Yanqi Wu, Hui Chen, Ying Tian, Zhenqi Yang, Duan Feng, Jiaqi Liu, Dan Wu","doi":"10.1166/jbn.2023.3553","DOIUrl":"https://doi.org/10.1166/jbn.2023.3553","url":null,"abstract":"N-Glycosylation is one of the most common and important protein posttranslational modifications. Structural aberrations of the N-glycans branching from glycoproteins are closely related to various disease occurrences and progressions. Therefore, global systematic identification of disease-related\u0000 N-glycans not only largely facilitates the understanding of their cellular functions but also promotes the development of new diagnostic and therapeutic biomarkers. However, N-glycans are low in abundance and hydrophilic, thereby hindering high-throughput, and large-scale N-glycan analysis\u0000 and N-glycoproteome research. Here, a zwitterion-functionalized polymer brush-grafted silica hybrid hydrophilic material (poly-SBMA-S) was fabricated by in situ growth of polymeric [2-(methacryloyloxy)ethyl]dimethyl(3-sulfopropyl)ammonium hydroxide (SBMA) chains on porous silica particles\u0000 through a surface-initiated atom transfer radical polymerization technique, and it was used as a new stationary phase for highly selective N-glycan enrichment. Human plasma demonstrated 64 N-glycans due to the densely packed polyzwitterion chains and the significantly increased coverage of\u0000 hydrophilic binding sites, demonstrating the high potential of the new hydrophilic matrix in the profiling and analysis of N-glycans and other hydrophilic targets.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48647579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Crucial Biological Roles of Long Noncoding RNA-Mediated Pyroptosis in Urological Tumors 长链非编码rna介导的焦亡在泌尿系统肿瘤中的重要生物学作用

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3637

Jiaogui Xie, Ying Jiang, Yichao Wang, Dalin He

引用次数: 0

A Convergent Fabrication of Silk Fibroin Coated Magnesium Oxide Conjugated Phosphate Functionalized Chitosan Hydrogel for Bone Regeneration 丝素包被氧化镁结合磷酸功能化壳聚糖骨再生水凝胶的聚合制备

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3652

Xi Li, Yong-tao Yi, P. Guo, Ru-dan Zhou, Hua Li

引用次数: 0

Green Synthesis of Magnetic Nanoparticles from Oldenlandia diffusa: Characterization and Therapeutic Activities 绿色合成白花玉兰磁性纳米颗粒:表征及治疗活性

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3660

Yingying Kong, Periyannan Velu, A. Vijayalakshmi, Chao Yuan

{"title":"Green Synthesis of Magnetic Nanoparticles from Oldenlandia diffusa: Characterization and Therapeutic Activities","authors":"Yingying Kong, Periyannan Velu, A. Vijayalakshmi, Chao Yuan","doi":"10.1166/jbn.2023.3660","DOIUrl":"https://doi.org/10.1166/jbn.2023.3660","url":null,"abstract":"This study investigated green synthesis of magnetic nanoparticles (MNP) using Oldenlandia diffusa, its cauterization, and therapeutic propertieson liver cancer cells via an in-vitro approach. Anti-cancer, anti-oxidant, and anti-diabetic properties of MNP were evaluated.\u0000 Synthesis of MNP was confirmed via UV-visible spectroscopy. Oldenlandia diffusa MNP (20–40 nm) showed absorption peak at 327 nm in UV-visible, XRD, EDX and FTIR spectral analysis. MNP were monoclinic, and rhombohedral with two stretching peaks at 646 cm−1, and\u0000 1098 cm−1 confirming Fe–O–Fe mode and Fe = O due to the iron element. IC50 values for MNP (IC50 74 μg/ml & 58 μg/ml) using DPPH and ABTS methods were high as compared to aqueous leaf extract of Oldenlandia diffusa\u0000 confirming anti-oxidant properties. MNP exhibited α-amylase inhibitory activity with maximum inhibition of 82.17±0.82 μg/ml for α-glucosidase inhibitory action of 84.82±0.89 μg/ml. MNP showed dose-dependant in-vitro tumor suppression\u0000 activity against HepG2 cells (16.08% at 150 μg/ml and 28.31% at 150 μg/ml). MNP also demonstrated anti-leishmanial activity at 400 g/ml (74.26±0.82 g/ml for promastigotes and 69.40±0.69 μg/ml for amastigotes). Oldenlandia diffusa actively\u0000 bonded with magnetic Fe-nanoparticles and suppressed the growth of HepG2 cells. It demonstrated anti-cancer, antioxidant, anti-leishmanaland anti-diabetic activities. Thus, Oldenlandia diffusa is a potential anti-cancer therapeutic drug for liver cancer.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47387858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Homeobox D9 Promotes Malignant Progression of Non-Small Cell Lung Cancer Through Negatively Regulating Kallikrein Related Peptidase 9 Homeobox D9通过负调控激肽释放酶相关肽酶9促进非小细胞肺癌的恶性进展

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3664

Juntao Hao, Zengqiang Shen, Junjun Ma

{"title":"Homeobox D9 Promotes Malignant Progression of Non-Small Cell Lung Cancer Through Negatively Regulating Kallikrein Related Peptidase 9","authors":"Juntao Hao, Zengqiang Shen, Junjun Ma","doi":"10.1166/jbn.2023.3664","DOIUrl":"https://doi.org/10.1166/jbn.2023.3664","url":null,"abstract":"We investigated the association between HOXD9 expression and prognosis in non-small cell lung cancer (NSCLC) and explored its underlying mechanisms in tumor progression. HOXD9 expression was examined in 42 pairs of NSCLC tissue samples using qRT-PCR, and its effects were evaluated in\u0000 NSCLC cell lines through lentiviral vector transfection, assessing cell growth and migration using CCK-8 and Transwell assays. Bioinformatics analysis identified KLK9 as a target gene of HOXD9, and their interaction was investigated in vitro and in vivo. HOXD9 mRNA expression\u0000 was significantly higher in NSCLC tissues compared to adjacent tissues, positively correlating with metastasis incidence and poor prognosis. HOXD9 knockdown reduced NSCLC cell migration and proliferation, while overexpression had the opposite effect. Luciferase assays confirmed HOXD9′s\u0000 binding to KLK9, leading to negative regulation of KLK9 expression. Knockdown of KLK9 reversed the inhibitory effect of HOXD9 on NSCLC cell migration and proliferation, while co-overexpression of KLK9 and HOXD9 had the opposite effect. Our findings demonstrate the close association of high\u0000 HOXD9 expression in NSCLC tissues with metastasis incidence and poor prognosis. HOXD9 likely exerts its biological function in NSCLC by negatively regulating KLK9. These results provide valuable insights into the molecular mechanisms driving NSCLC progression, highlighting HOXD9 as a potential\u0000 prognostic biomarker and therapeutic target.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47593813","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Ultrasound-Promoted Nanoreagent for Homologous Targeted Synergistic Chemodynamic/Sonodynamic Tumor Therapy 超声促进纳米试剂用于同源靶向协同化疗/声动力肿瘤治疗

IF 2.9 4区医学

Journal of biomedical nanotechnology Pub Date : 2023-08-01 DOI: 10.1166/jbn.2023.3655

Huiying Zuo, Hui Fang Xiao, Deqiang Wang, Ranran Wang

{"title":"Ultrasound-Promoted Nanoreagent for Homologous Targeted Synergistic Chemodynamic/Sonodynamic Tumor Therapy","authors":"Huiying Zuo, Hui Fang Xiao, Deqiang Wang, Ranran Wang","doi":"10.1166/jbn.2023.3655","DOIUrl":"https://doi.org/10.1166/jbn.2023.3655","url":null,"abstract":"The establishment of a tumor theranostic platform with high efficacy is attracting considerable interest in cancer treatment. Nevertheless, considerable obstacles need to be overcome in developing such a platform. In this study, a versatile theranostic system (CM-FMNPs-Cur) was produced\u0000 utilizing MnO2 nanoflowers, which provided targeted and synergistic therapy for cancer through the incorporation of sonodynamic and chemodynamic therapies. By integrating Fe3O4, MnO2, and curcumin, CM-FMNPs-Cur manifests a substantial amount of reactive\u0000 oxygen species toward deep-seated cancers. As a sonosensitizer, curcumin was used for chemodynamic therapy (CDT)_and sonodynamic therapy (SDT). In the acidic environment of malignant cells, CM-FMNPs-Cur can decompose into Mn2+ and Fe2+, triggering a Fenton-like reaction\u0000 and generating ·OH radicals and oxygen. Ameliorating tumor hypoxia also enhanced the effectiveness of SDT in neoplasms. The homologous targeting effect of CM-FMNPs-Cur on cancer cell membranes led to its accumulation in the tumor region. Here, the mutual promotion of CDT and SDT resulted\u0000 in an amplified therapeutic outcome. All these results revealed the vast potential of CM-FMNPs-Cur for secure and highly effective collaborative tumor treatment, indicating future clinical applications.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48859292","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0