Nanoscience and Nanotechnology Letters最新文献

{"title":"The Growth Inhibition of Targeted Pluronic/Formononetin Nanocomposite on Oral Squamous Cell Carcinoma In Vitro","authors":"Ming Liu, Chen Lin, Xiaoqing Huang, Yuxiang Lin","doi":"10.1166/NNL.2020.3195","DOIUrl":"https://doi.org/10.1166/NNL.2020.3195","url":null,"abstract":"Natural flavonoid formononetin (FN) has anticancer effects, but the hydrophobic structure, characteristics of the short half-life in vivo, limiting its clinical wide-ranging application. In this study, FN loaded Pluronic (PF)@folic acid (FA) micelles (FN-PF@FA), were prepared\u0000 to improve the solubility, bioavailability and targeting. FA coupling PF was prepared by carbodiimide crosslinker chemical method, FN-PF@FA micelles were prepared by modified film hydration method, and compared the antitumor activity of FN loaded micelles with free FN In Vitro. The\u0000 spherical smooth surface of FN-PF@FA micelles had smaller particle size (112.3±5.3 nm), high encapsulation efficiency (86.14±2.68%), high negative zeta potential (-25.8±0.57 mV), low critical concentration CMC (0.03 mg/mL), and better sustained release profile. In addition,\u0000 FN-PF@FA micelles have a positive targeting effect on oral squamous cell carcinoma cells (SCC3). In 48 hours, the growth inhibition of 50% (GI50) was 28.6±1.2 μg/mL for FN and 17.4±0.78 μg/mL for FN-PF, the dose dropped by nearly 38.46%. In addition, the GI50\u0000 value of FN-PF@FA was 9.5±0.3 μg/mL, 66.43% lower than FN and 44.83% lower than FN-PF. Furthermore, the laser scanning confocal microscopy revealed that the conjugation of FA significantly improves the active targeting ability of micelles. FN-PF@FA micelles have the potential\u0000 to target the release of anticancer drugs with higher bioavailability, further provides a new avenue for the application of traditional Chinese medicine extract in oral malignant tumor.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1022-1029"},"PeriodicalIF":0.0,"publicationDate":"2020-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45305992","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Automatic Segmentation Method of Breast Tumor Ultrasonic Images Based on Attention-Enhancing Unet","authors":"Yu Yan, X. Cai, Ge Fang, Wei Zhu, Jian Liu, Funan Xiao, Manxue Zhao, Wang Zuming, Yiyun Wu","doi":"10.1166/NNL.2020.3201","DOIUrl":"https://doi.org/10.1166/NNL.2020.3201","url":null,"abstract":"In order to improve the accuracy of the segmentation of the breast ultrasound image lesion, Attention-Unet was improved, and an Attention-enhancing Unet (AE-Unet) model is proposed. First, the network loss function was improved. Based on the output value of the traditional network end,\u0000 output weights of all attention gate were integrated. Compared with the standard lesion template, it was used to obtain accurate network loss values; Secondly, the network training method was improved, and the strategy of combining thickness and fineness was adopted. The overall loss function\u0000 was used to train the overall network to make the network basically stable; then the partial loss function was used to alternately train the backbone network and the attention gate module in turn. Fine-tuning was used to further improve the accuracy of network parameters. The combination of\u0000 the two greatly improves the accuracy of segmentation of the breast ultrasound lesion area. The experimental results on the breast ultrasound data actually collected in the hospital show that the proposed AE-Unet model has an M-IOU of 81.24%, precision of 85.88%, F1 of 80.58%, Acc of\u0000 93.85% and specificity of 97.48%, PPV is up to 85.88%, which has achieved better segmentation results than existing advanced algorithms.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"996-1005"},"PeriodicalIF":0.0,"publicationDate":"2020-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41711299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fenofibrate Improves the Progression of Diabetic Retinopathy by Regulating miR-21-IL6R Axis","authors":"Yan Zhang, Bihua Xie","doi":"10.1166/NNL.2020.3207","DOIUrl":"https://doi.org/10.1166/NNL.2020.3207","url":null,"abstract":"Diabetic retinopathy (DR) is a kind of irreversible visual impairment common in adults. Several studies have confirmed the ability of fenofibrate (FNB) to mitigate DR pathology. miR-21 has also been shown to improve the hemodynamics of DR model rats. Presently, it is not clear whether\u0000 FNB can play a therapeutic role in DR via regulating miR-21, and we will supplement this gap through this research. First, in this research, we constructed an in vitro model of DR by inducing ARPE-19 cells with high glucose (HG). After successful induction, the proliferation ability\u0000 of ARPE-19 cells was inhibited, the apoptosis ability was enhanced, and the oxidative stress index (reactive oxygen species, ROS) level was increased. Subsequently, we discovered that the cell vitality was significantly improved, the apoptosis ability was inhibited, and oxidative stress injury\u0000 was improved under FNB intervention in the in vitro model of DR. In mechanism, FNB can improve the viability of DR model in vitro and alleviate the oxidative stress injury induced by HG through down-regulating miR-21 or up-regulating IL-6R. Furthermore, miR-21 has a targeted\u0000 regulatory relationship with IL6R, which can negatively regulate the level of IL6R. Based on the above, we conclude that FNB can improve the viability and alleviate oxidative stress injury of DR model in vitro via regulating the miR-21-IL6R axis.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"961-969"},"PeriodicalIF":0.0,"publicationDate":"2020-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48056214","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correlation Between Intestinal Microflora and Gastrointestinal Dysfunction in Elderly Patients with Sepsis","authors":"Hailing Yang, Zhang Xiaolin, Chen Weijun, Libo Shang, Bin Chen, Pang Yongcheng, Zhang Guilan, Wan Qing, Chang Zhang, Jianqin Zhou, Rui Lv, Shi Xingqian, Mingyan Li","doi":"10.1166/NNL.2020.3206","DOIUrl":"https://doi.org/10.1166/NNL.2020.3206","url":null,"abstract":"Multiple organ dysfunction syndrome (MODS) is a key factor that leads to death in elderly patients with sepsis. Therefore, early prevention and treatment of gastrointestinal dysfunction (GIDF) in elderly patients with sepsis is an important measure to prevent MODS occurrence. This research\u0000 explores the correlation between intestinal microflora and GIDF in elderly patients with sepsis and provides ideas for the prevention and treatment of GIDF in elderly patients with sepsis. In this study, 152 patients with sepsis (122 patients with sepsis and GIDF) treated in the Third Affiliated\u0000 Hospital of Yunnan University of Chinese Medicine from January to September 2019 were selected as the sepsis group and 100 elderly who had normal physical examination results were selected as the control group. The common intestinal microflora of the two groups was compared. Patients with\u0000 sepsis and GIDF were treated as the GIDF group and the other patients with sepsis were treated as the non-GIDF group. The common intestinal microflora, gastrointestinal indicators, serum inflammatory factors, and immune function indices were compared between the two groups. Correlation analysis\u0000 of the observed indices with statistical significance was carried out. The results showed 152 patients with sepsis and 122 patients with sepsis and GIDF; thus, the incidence of sepsis with GIDF was 80.26%. The total average score of sepsis with GIDF was 3.61±0.09. There was no statistically\u0000 significant difference in GIDF scores of patients ages 65–75 and > 75 years old. The number of Bifidobacterium and Lactobacillus in elderly patients with sepsis was lower and the number of Escherichia coli was higher than in the control group. In elderly patients\u0000 with sepsis, the number of Bifidobacterium and Lactobacillus in the GIDF group was lower and the number of E. coli was higher than in the non-GIDF group. White blood cell (WBC) count, procalcitonin (PCT), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α),\u0000 gastrin (GAS), and diamine oxidase (DAO) in GIDF patients were higher and motilin (MOT), CIT (CIT), CD4+, and CD8+ were lower than in the non-GIDF group. WBC count, PCT, CRP, TNF-α, GAS, and DAO were negatively correlated with the number of Bifidobacterium\u0000 and Lactobacillus but positively correlated with E. coli. MOT, CIT, CD4+, and CD8+ were positively correlated with the number of Bifidobacterium and Lactobacillus but negatively correlated with E. coli. There was a negative correlation\u0000 between Bifidobacterium and Lactobacillus and GIDF score and a positive correlation between E. coli and GIDF score. Therefore, the change in the intestinal microflora in elderly patients with sepsis is related to GIDF.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1030-1037"},"PeriodicalIF":0.0,"publicationDate":"2020-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45050885","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Long-Chain Non-Coding RNAs Metastasis-Associated Lung Adenocarcinoma Transcript-1 Is Involved in Growth of Nasopharyngeal Carcinoma Cells by Targeting miR-205-5p","authors":"Yuetang Mi, Haiyan Li, Jingchuan He","doi":"10.1166/NNL.2020.3204","DOIUrl":"https://doi.org/10.1166/NNL.2020.3204","url":null,"abstract":"The incidence of nasopharyngeal carcinoma (NC) has been rising. The prognosis of NC remains unsatisfactory; therefore, it is necessary to find new ways to improve it. LncRNA MALAT1 (MALAT1) is a well-studied gene, but its status in NC remains unclear. More experimental analyses are\u0000 needed to uncover the role of MALAT1 in NC for a reliable and accurate theoretical basis for NC’s diagnosis and treatment. First, NC patients admitted to our hospital from February 2018 to March 2020 and healthy controls who underwent physical examinations during the same period were\u0000 enrolled in this study. MALAT1 and miR-205-5p in the patients’ peripheral blood and tissues were detected. The expression of MALAT1 was high, and the expression of miR-205-5p was low in the NC patients. Both genes were effective in predicting the occurrence of NC, and their expression\u0000 was negatively correlated. According to in vitro experiments, inhibiting MALAT1 and increasing miR-205-5p could reduce CNE-2Z cells’ proliferation and increase their apoptotic rate. However, increasing MALAT1 and inhibiting miR-205-5p had opposite effects. Through the online website\u0000 ENCORI, complementary sites were found in MALAT1 and miR-205-5p. According to dual-luciferase reporter gene assay, MALAT1-WT’s luciferase activity was inhibited by the co-transfection of miR-205-5p-min, while MALAT1-MUT’s luciferase activity was enhanced by the co-transfection\u0000 of miR-205-5p-inh. Lastly, through rescue experiments, we found no difference in the biological behavior between the cells in the MALAT1 si-RNA+miR-205-5p-inh group and the MALAT1+NC-RNA group. Therefore, MALAT1 promotes NC cell proliferation, inhibits apoptosis, and participates in NC’s\u0000 development via specifically regulating miR-205-5p.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"970-978"},"PeriodicalIF":0.0,"publicationDate":"2020-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45317344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis of Gold Nanoparticles in Cyclodextrin-Tetrachloroaurate System","authors":"Y. S. Pestovsky, T. Srichana","doi":"10.1166/NNL.2020.3176","DOIUrl":"https://doi.org/10.1166/NNL.2020.3176","url":null,"abstract":"Tetrachloroaurate reduction with α-, β-, and γ-cyclodextrin at pH 10.56 was studied in this study by dual-angle dynamic light scattering and spectrophotometry. The nanoparticles were also characterized by scanning electron microscopy. In contrast\u0000 with our previous study, the nanoparticles were purified by centrifugation prior to characterization. The reaction is considered to be a promising one-step method for preparation of gold nanoparticles with immobilized cyclodextrins without the need for seeding. Unlike in our previous study,\u0000 the reaction mixture was boiled under reflux conditions instead of an open vial. This change sped up the reaction, and extensive aggregation was avoided. For the first time, this study demonstrated that all three cyclodextrins were able to reduce tetrachloroaurate at room temperature as well,\u0000 but long incubation periods were necessary. This is the first publication discussing the mechanism of tetrachloroaurate reduction by cyclodextrins. The first stage of the reaction involved tetrachloroaurate hydrolysis despite presence of hydrochloric acid in the commercial preparation of tetrachloroauric\u0000 acid (HAuCl4). Tetrachloroaurate was therefore the precursor but not the actual oxidant. The previous hypothesis on autocatalytic decarboxylation of cyclodextrins was therefore proven wrong. Particle growth was accompanied by nucleation, leading to coexistence of nanoparticles and\u0000 nanoclusters. The resulting nanoparticles contained a small fraction of aggregates, probably because of sodium chloride generated from hydrochloric acid. However, besides playing this detrimental role, chloride ions facilitated nucleation by stabilizing gold intermediates. The nanoparticles\u0000 solutions could be stored at least for three months, which contrasted with our previous work, where the nanoparticles were stable for only three days. For the first time, freeze-drying and reconstitution of the resulting nanoparticles were investigated, and they were shown to be free from\u0000 aggregation. The synthesized gold nanoparticles are recommended for use as drug carriers.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"849-862"},"PeriodicalIF":0.0,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41573273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Study on the Immunomodulatory Effect of Quercetin Nanoparticles Loaded with Chitosan on a Mouse Model of Ovalbumin-Induced Food Allergy","authors":"Hua Feng, Xiujuan Xiong, Qunying Xu, Zhongwei Zhang, J. Feng, Yongning Wu","doi":"10.1166/NNL.2020.3197","DOIUrl":"https://doi.org/10.1166/NNL.2020.3197","url":null,"abstract":"As a common immune disease, food allergy has become a difficult problem for public health organizations. Food allergies are often accompanied by skin reactions, allergic rhinitis, gastrointestinal diseases, and other complications which can induce anaphylactic shock and even death.\u0000 However, mechanisms underlying food allergies are not fully understood. Therefore, exploring the mechanism of induction of food allergy is a critical first step toward discovering a future treatment. Here we constructed a quercetin–chitosan nanoplasmid (QT–CS NPs), which can regulate\u0000 the balance of Th1/Th2 cells, and used it to treat ovalbumin (OVA)-induced allergies in mice. The results suggest that compared to natural quercetin, the cytotoxicity of QT–CS NPs was weakened. QT–CS NPs also increased the expression of the Th1 cytokine INF-γ, reduced\u0000 the secretion of Th2 cytokine IL-4, regulated the balance drift of Th1/Th2 cells in allergic mice, and demonstrated an excellent immunomodulatory effect. Therefore, QT-CS NPs can provide new treatment ideas for the prevention and treatment of food allergy, and this study can lay an experimental\u0000 foundation for the clinical application of QT-CS NPs.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"915-920"},"PeriodicalIF":0.0,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44837181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Abnormal Platelet miRNA Profile Associated with Endometriosis","authors":"Miaomiao Du, Jiabao Wu, Yali Zhu, D. Mao, Zhihong Liu, Guifang Gong, Qinsheng Lu, Yan-qing Huang","doi":"10.1166/NNL.2020.3184","DOIUrl":"https://doi.org/10.1166/NNL.2020.3184","url":null,"abstract":"Endometriosis (EM) is a condition characterized by the growth of functional endometrium in areas other than the uterus. In this study, microRNAs (miRNAs) were extracted from peripheral blood platelets from patients diagnosed with EM and from healthy controls for high-throughput sequencing;\u0000 differential expression of miR-542-3p and miR-618 was identified via database comparisons. Dual pathway enrichment analysis revealed that both miRNAs may target the cyclin protein, G1/S-specific cyclin-D2 (CCND2). CCND2 was verified as a target gene of both miR-542-3p and miR-618 via experiments\u0000 carried out in vivo. Taken together, these results reveal that CCND2 may serve as a functional biomarker for the diagnosis of EM.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"863-873"},"PeriodicalIF":0.0,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43310978","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Screening and Postoperative Evaluation of Breast Cancer Based on Serum Proteins Analysis Using Label-Free Surface-Enhanced Raman Spectroscopy Technology","authors":"Xiaokun Liu, Xueliang Lin, Zongwei Huang, Youzhi Zhu, Min Peng, Xianggang Jia, Shuyun Weng, Y. Weng, S. Feng","doi":"10.1166/NNL.2020.3189","DOIUrl":"https://doi.org/10.1166/NNL.2020.3189","url":null,"abstract":"Breast cancer is a malignant tumor of breast epithelial tissue. Early diagnosis and postoperative evaluation of breast cancer are critical to improve the survival rate. The current main screening methods are mammography and computerized tomography (CT), however, these methods suffer\u0000 from false positives, over-diagnosis and radiation risk. Herein, unlabeled surface-enhanced Raman spectroscopy (SERS) technology combined with silver nanoparticles that was used to measure and analyze peripheral serum protein samples from patients of breast cancer for preoperation, postoperation\u0000 and normal subjects. Results showed that there were significant differences in the serum protein SERS spectra among three groups due to changes in certain biochemical compositions related to breast cancer transformation. Moreover, diagnostic sensitivity, based on principal component analysis\u0000 combined with linear discriminant analysis (PCA-LDA) for pre-surgery versus post-surgery, post-surgery versus normal and pre-surgery versus normal were 96.7%, 53.3%, and 100%, respectively, and the diagnostic specificities were 96.7%, 46.7%, and 96.7%, respectively. Therefore, serum protein\u0000 SERS combined with PCA-LDA analysis holds promising potential as a novel strategy for early screening and postoperative evaluation of breast cancer.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"901-908"},"PeriodicalIF":0.0,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42200995","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Coarse-to-Fine Registration on 3D Multi-Phase Abdominal CT Images","authors":"Shao-di Yang, Fan Zhang, Zhen Yang, Xiaoyu Yang, Shuzhou Li","doi":"10.1166/NNL.2020.3194","DOIUrl":"https://doi.org/10.1166/NNL.2020.3194","url":null,"abstract":"Registration is a technical support for the integration of nanomaterial imaging-aided diagnosis and treatment. In this paper, a coarse-to-fine three-dimensional (3D) multi-phase abdominal CT images registration method is proposed. Firstly, a linear model is used to coarsely register\u0000 the paired multiphase images. Secondly, an intensity-based registration framework is proposed, which contains the data and spatial regularization terms and performs fine registration on the paired images obtained in the coarse registration step. The results illustrate that the proposed method\u0000 is superior to some existing methods with the average MSE, PSNR, and SSIM values of 0.0082, 21.2695, and 0.8956, respectively. Therefore, the proposed method provides an efficient and robust framework for 3D multi-phase abdominal CT images registration.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"909-914"},"PeriodicalIF":0.0,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43495770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}