Zhi Ji, Xu Liu, Xia Wang, Yuan Ren, Ying Liu, Shuangyu Han, Jingkang Zhao, Xingchun Gou, Yuan He
{"title":"Changes in Mitochondrial Morphology and Mitochondrial Fission/Fusion Gene Expression in Retinal Pigment Epithelial Cells During Oxidative Stress","authors":"Zhi Ji, Xu Liu, Xia Wang, Yuan Ren, Ying Liu, Shuangyu Han, Jingkang Zhao, Xingchun Gou, Yuan He","doi":"10.1166/NNL.2020.3229","DOIUrl":"https://doi.org/10.1166/NNL.2020.3229","url":null,"abstract":"Age-related macular degeneration (AMD) represents a serious impairment for the elderly. Because the pathogenesis of AMD has not been completely defined, the available therapeutic treatments are not ideal. Retinal pigment epithelial (RPE) cells are essential for photoreceptor cell maintenance\u0000 and survival; however, the mechanisms underlying RPE cell damage and AMD remains to be elucidated. It is known that abnormal mitochondrial gene expression causes mitochondrial dysfunction, induces cell damage, and results in disease. In this study, ARPE-19 cells were treated with different\u0000 concentrations of H2O2. It was found that excessive H2O2 concentration resulted in significant contraction of ARPE-19 cells and increased cell death, and destruction of mitochondrial structure as well as membrane and crest. RT-PCR results showed\u0000 that decreased expression of the Fis1 gene was evident in H2O2-treated cells. There were no significant differences observed among the different H2O2 concentration groups. The expression of the fission genes, MTP18 and Dnmp1, and the fusion genes,\u0000 Mnf1 and Mnf2, was not significantly different. Real-time PCR results revealed that the expression of the Fis1 gene decreased concomitantly with different concentrations of H2O2, whereas the expression of the Mfn2 gene increased by treatment with 200 μMH2O2.\u0000 There were no significant differences in the expression of the other genes. These results indicate that abnormal expression of the mitochondrial Fis1 fission gene, and the Mfn2 fusion gene caused mitochondrial dysfunction in ARPE-19 cells. This indicates that the imbalance of mitochondrial\u0000 dynamics may contribute to cell death in an oxidative stress environment.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1192-1199"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42703711","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":"Analysis of Nano-Silver Application in Nasosinusitis and Its Effect on Eotaxin","authors":"Zhiguo Ouyang, Xiaohong Wu","doi":"10.1166/NNL.2020.3228","DOIUrl":"https://doi.org/10.1166/NNL.2020.3228","url":null,"abstract":"Nasosinusitis (NA) is a severe inflammatory disease. In recent years, NA’s morbidity and its threat to patients is becoming increasingly serious. Clinically, it is considered that NA recurrence and refractoriness is due to eosinophil (EO) proliferation. Nanotechnology improvement\u0000 and application is a breakthrough in diagnosing and treating clinical diseases. Among which, nano-silver (NNS) technology is consistent with effective NA treatment, but there is no reliable research demonstrating this. In this study, inflammatory factors, EO chemokines, and epidermal growth\u0000 factors (EGF) in NNS-treated NA mice were analyzed to determine whether NNS had a potential therapeutic effect on NA. The results showed the clinical NA symptom score in mice decreased (P <0.05). Inflammatory factors in nasal lavage fluid (NLF), as well as NF-κB and\u0000 eotaxin in the maxillary sinus mucosa were inhibited (P <0.05). Finally, EGF and epidermal growth factor receptor increased (P <0.05). These data demonstrate that NNS is an effective NA treatment. NNS may be able to reduce the cell inflammation occurrence and development\u0000 by inhibiting EO chemokine expression, and promoting certain repair programs to the damaged nasal mucosa. There, NNS may become a feasible scheme for future NA treatment.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1151-1158"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42039311","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}
Junhua Yu, Jianping Fan, Yujun Zhao, Hang-Qing Lu, J. Qian
{"title":"Gold Nanoparticle-Delivered miR-26a Restricts Growth and Invasion of Gastric Cancer Cells","authors":"Junhua Yu, Jianping Fan, Yujun Zhao, Hang-Qing Lu, J. Qian","doi":"10.1166/NNL.2020.3222","DOIUrl":"https://doi.org/10.1166/NNL.2020.3222","url":null,"abstract":"This paper aims to estimate the effect of miR-26a delivered by gold nanoparticles (AuNPs) on gastric cancer (GC) cell growth and invasion. In vitro cultured GC cell strains SGC-7901 and MGC- 803 were processed for transfection of miR-26a-mimic, miR-NC, AuNP-ctrl, and AuNP-miR-26a\u0000 groups. MTT, Transwell, and flow cytometry were employed to evaluate the groups’ impact on cell vitality, invasion, and apoptosis. Western blotting (WB) was used to quantify the levels of apoptosisrelated proteins. In addition, solid tumors in mice were monitored for any changes that\u0000 occurred under treatment by each group. We observed that overexpressing miR-26a restricted the vitality and invasion, and promoted apoptosis of GC cells; this effect became more significant with the injection of AuNP-delivered miR-26a, which evidently suppressed the growth of tumors in\u0000 vivo. We conclude that, miR-26a serves as a tumor suppressor gene (TSG) in GC, and its effectiveness may be enhanced through delivery by AuNPs.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1226-1232"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44303864","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":"Nano Magnetic Beads Combined with Polymerase Chain Reaction Detection of 2019-nCoV and Analysis of the Epidemiological and Clinical Characteristics of 2019-nCoV Patients","authors":"Lingyan Fan, Linsong Liu, Nanhong Zheng, Dezhou Li, Hong-bin Tu, Hong-fei Xuan, Donghui Wang, Wei Wang, He-ming Chen, Honghua Ye, Yao-ren Hu, Shun Zhang, Gang Yuan","doi":"10.1166/NNL.2020.3231","DOIUrl":"https://doi.org/10.1166/NNL.2020.3231","url":null,"abstract":"The 2019 novel coronavirus (2019-nCoV) is extremely infectious and the human population globally is generally susceptible to it Its spread poses a serious threat to public health Analyzing the epidemiological and clinical characteristics of patients with coronavirus disease 2019 (COVID-19) can more fully demonstrate its characteristics and evolution, and provide effective scientific strategies for preventing and controlling the epidemic In this study, nucleic acid detection was performed on 228 suspected COVID-19 cases in Ningbo City from January 2020 to August 2020 The RNA of the virus was extracted by the nano magnetic bead method and was detected by real-time fluorescent quantitative PCR Patients whose test results were positive were considered as research subjects Clinical data of the patients were collected through a medical record system and the epidemiology and characteristics of COVID-19 were analyzed Among the 228 suspected cases, 108 patients had a positive nucleic acid test result The 108 confirmed patients were selected as research subjects, including 9 patients with mild symptoms, 85 patients with common symptoms, and 14 patients with severe symptoms The age range of the patients was 17-82 years Overall, 35 patients (32 40%) had complications The median incubation period for patients was 7 5 days Seventy patients (64 81%) had a contact history with a diseased individual In terms of the clinical symptoms of the patients, the following symptoms were identified: fever in 92, cough in 62, fatigue in 34, sputum expectoration in 32, dry cough in 22, sore throat in 19, diarrhea in 8, headache in 8, shortness of breath in 7, nasal congestion in 5, and muscle ache in 5 cases The results of chest CT imaging showed that 96 patients (88 89%) exhibited bilateral or unilateral pneumonia lesions of varying degrees, manifested as bilateral or unilateral lung patches, cloudy floccules, patches with an increased density shadow The CT images of 12 patients (11 11%) showed no obvious abnormalities, while 24 cases (22 22%) had absolute white blood cell count of 10x10(9)/L The absolute neutrophil count in 10 patients (9 26%) increased (>6 3x10(9/)L) The absolute lymphocyte count in 49 patients (45 37%) decreased (10 mg/L);14 (12 96%) had elevated D-dimer (>256 mu g/L);14 (12 96%) had high levels of alanine aminotransferase (>40 U/L);9 (8 33%) had increased aspartate aminotransferase (>40 U/L);while 32 (29 6%) had increased lactate dehydrogenase (>250 U/L) COVID-19 in Ningbo is mainly transmitted in clusters, and elderly patients are more likely to develop severe symptoms There is no significant difference in clinical characteristics between patients with severe conditions and those with mild and common ones","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1207-1214"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45655174","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}
Qitong Liu, Li Wang, Huajun Jiang, Tong Chen, Weiguo Zhang, Tao Jiang
{"title":"Application of Magnetic Nano-MR Molecular Imaging Technology in the Relationship Between Meniscus Injury and Knee Joint Movement","authors":"Qitong Liu, Li Wang, Huajun Jiang, Tong Chen, Weiguo Zhang, Tao Jiang","doi":"10.1166/NNL.2020.3230","DOIUrl":"https://doi.org/10.1166/NNL.2020.3230","url":null,"abstract":"Knee osteoarthritis (KOA) can lead to joint malformations and dysfunction. The meniscus, as the fibrous cartilage of the medial and lateral articular surfaces of the tibia, functions in load transfer, shock absorption, and knee joint stabilization. Injury of the meniscus reduces its\u0000 protective effect on the knee joint and increases the risk of KOA. Early detection of meniscus injury and accurate assessment of its severity could help to clinically formulate corresponding treatment plans, delay disease progression, and reduce the risk of KOA. Magnetic resonance imaging\u0000 (MRI) can clearly characterize the tissue structure of the meniscus in all directions, which is convenient for observing the damage to it. However, the sensitivity of traditional MRI to cartilage tissue is still limited. Superparamagnetic iron oxide nanoparticles (SPION), as a nano-biomaterial\u0000 with superparamagnetic properties, can enhance MR signal intensity and imaging sensitivity. The use of SPION as a contrast agent for MRI diagnosis of meniscus injury may help to improve the diagnostic accuracy. In this study, the results of arthroscopy were used as a gold standard to compare\u0000 the diagnostic value of traditional MRI and MRI+SPION in meniscus injury. We found that SPION could significantly improve the sensitivity, specificity, and accuracy of MRI in diagnosing meniscus injury. According to the classification of the severity of meniscus injury by MRI+SPION, six degrees\u0000 of freedom of knee joints were compared in each group, and major differences were found in each degree of freedom of knee joints in the patients with different severity grades. The more severe the injury, the smaller the range of degrees of freedom of the knee. The above results show that\u0000 the knee joint activity of patients with meniscus injury can be evaluated according to magnetic nano-magnetic resonance molecular imaging technology, and the corresponding clinical treatment plan can be made according to the diagnosis and evaluation results.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1200-1206"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49298147","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}
Ye Shen, Lianming Zhou, Meiyu Xu, Zhanhai Tan, K. Yao, Guanghui Zhu
{"title":"Killing Action of Cuprous Oxide Nanoparticles on Breast Cancer Cells","authors":"Ye Shen, Lianming Zhou, Meiyu Xu, Zhanhai Tan, K. Yao, Guanghui Zhu","doi":"10.1166/NNL.2020.3223","DOIUrl":"https://doi.org/10.1166/NNL.2020.3223","url":null,"abstract":"Breast cancer (BC), a common malignancy, is intractable at present. Cuprous oxide nanoparticles (Conps) are new nanoparticles crucial in inhibiting tumor progression according to early studies, but their role in BC remains unclear. Field emission-scanning electron microscopy (FESEM)\u0000 and field emission–transmission electron microscopy (FETEM) were used for the morphological observation of Conps, and energy-dispersive X-ray spectroscopy was used for their elemental composition analysis. Multipoint nitrogen adsorption was used to determine the Brunauer–Emmet–Teller\u0000 of Conps, and their mean hydrodynamic size and zeta potential in water and intact cell medium were determined by dynamic light scattering (DLS). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Cell Counting Kit-8, Transwell assay, and flow cytometry were used to determine\u0000 the effects of Conps on the biological function of BC cells. Western blotting was used to determine the changes in apoptosis-related proteins. Using FESEM and FETEM based on DLS and zeta potential determination, Conps was successfully constructed. Conps significantly lowered the viability\u0000 of BC cells in a dose-dependent manner, significantly inhibited cell proliferation, invasion, and migration activities, and induced cell apoptosis. Western blotting demonstrated that Conps up-regulated Bax and caspase-3 proteins and down-regulated Bcl-2 protein. The results suggest that Conps\u0000 can suppress the proliferation, invasion, and migration of BC cells and accelerate their apoptosis. This is a new potential strategy for the clinical treatment of BC.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1233-1238"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42974456","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}
Bingshuang Xue, Yi Xue, Jiaojiao Zhou, Qichao Yang
{"title":"The Effect of Curcumin Nano-Suspension on Myocardial Fibrosis in Diabetic Rats","authors":"Bingshuang Xue, Yi Xue, Jiaojiao Zhou, Qichao Yang","doi":"10.1166/NNL.2020.3225","DOIUrl":"https://doi.org/10.1166/NNL.2020.3225","url":null,"abstract":"The occurrence of complications of diabetic patients not only increases the difficulty and burden of treatment but also significantly affects the health and safety of patients. Traditional therapeutic drugs are prone to drug resistance, which affects the therapeutic effect. In recent\u0000 years, the application of plant-derived natural compounds in the treatment of diseases has become a hot spot in the research of diabetes drugs. Curcumin has anti-tumor, anti-inflammation, anti-oxidation and antimicrobial effects, but the mechanism of its effect on cardiomyocytes in diabetic\u0000 patients is not yet clear. In this study, curcumin was prepared into nano-preparations and its mechanism of action in the process of myocardial fibrosis in diabetic rats was further explored. We found that injection of curcumin nano-suspension can increase the LVIDd and LVFS of rats, while\u0000 reducing the serum CKMB, LDH, AST and cTnI levels. Further exploration found that curcumin can reduce serum TNF- α and IL-1 β levels in diabetic rats, while increasing the SOD and GSH-Px activities of myocardial tissue, and reducing MDA content. These suggests that curcumin can\u0000 reduce inflammation and oxidative stress in diabetic rats. Therefore, this study believes that curcumin nano-suspension can effectively inhibit diabetic cardiomyocyte fibrosis, oxidative stress, and inflammation and protect the rat myocardium.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1215-1220"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46226651","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":"Using Artificial Neural Networks for Fractal Interpolation Approach in 3D","authors":"Rashad Al-Jawfi","doi":"10.1166/NNL.2020.3226","DOIUrl":"https://doi.org/10.1166/NNL.2020.3226","url":null,"abstract":"In this paper, we introduce a new method for fractal interpolation, herein called Neural Network Algorithm (NNA), which is based on Iterated Functions Systems (IFS); proposed to self-affine signals interpolation with error of expected interpolation. Experiments on theoretical data show\u0000 that the proposed interpolation schemes can obtain the expected point value and work with great precision in rebuilding the specified data profile, which leads to a significant advantage over other interpolation methods.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1221-1225"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45851123","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}
Xiaoyu Sun, Huanxin Wang, Xuan Gu, Zheng He, Lidan Xu, M. Shi
{"title":"Effects of Needle Flake Particles on Splitting Experiment of Asphalt Mixture: An Experimental and Numerical Investigation","authors":"Xiaoyu Sun, Huanxin Wang, Xuan Gu, Zheng He, Lidan Xu, M. Shi","doi":"10.1166/NNL.2020.3235","DOIUrl":"https://doi.org/10.1166/NNL.2020.3235","url":null,"abstract":"The purpose of this work is to study the influence of the content and shape of needle flake particles on the meso-scale splitting test performance of asphalt mixture, a three-dimensional laser scanning method is used to build the needle and flake particle model, and then the particle\u0000 model is put into the numerical simulation software EDEM to establish the splitting experimental model. The influence of loading speed and model parameters on the experimental results is revealed by assigning macro and micro parameters to different particles. With calibrating the parameters\u0000 by Marshall experiment, the simulation experiment results after parameter adjustment are in good agreement with the experimental results in laboratory. The results show that the splitting experimental model can be established intuitively by means of using 3D discrete element modeling technology,\u0000 and the stress of each particle size in the loading process can be observed directly. The loading speed, the content of needle flake particles, the size of needle flake particles and the distance from the loading area all have a great influence on the experimental results.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1166-1170"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44865761","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}
Xingshang Xu, Yuan Liu, Zhu Chen, Hui Chen, Yan Deng, Song Li, N. He
{"title":"Cardiac Troponin I Microfluidic Chip Driven by Adaptive Pressure","authors":"Xingshang Xu, Yuan Liu, Zhu Chen, Hui Chen, Yan Deng, Song Li, N. He","doi":"10.1166/NNL.2020.3244","DOIUrl":"https://doi.org/10.1166/NNL.2020.3244","url":null,"abstract":"To develop and design an adaptive microfluidic chip for accurate determination of cardiac troponin I (cTnI) in whole blood sample and explore the operating parameters of the chip in detecting cTnI, in order to provide a novel strategy for the detection of cTnI, cTnI microfluidic chip\u0000 was prepared by injection moulding, and the improved polystyrene polymer was used as the chip substrate to construct a three-layer composite structure, namely the upper, middle, and lower layers. The antihuman troponin I antibody I/II was grafted onto the chip surface to construct the detection\u0000 reaction zone using UV-induced production of surface-active free radicals. The stability of the chip preparation process, the running performance of the chip, and the analytical performance of the whole blood samples were investigated. It was shown that I adaptive pressure-driven microfluidic\u0000 chip has the advantages of easy bonding, integration, and a simple and stable production process. In the actual detection and analysis, the chip has high selectivity for cTnI in whole blood, lower detection limit (0.054 ng/mL), and small difference between batches (RSD% 2.50%). Therefore,\u0000 the chip is assumed to provide novel strategy for the assessment of myocardial infarction by detecting cTnI.","PeriodicalId":18871,"journal":{"name":"Nanoscience and Nanotechnology Letters","volume":"12 1","pages":"1239-1247"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48600544","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}