Chun-Feng Lu, Ye Dai, Yun Tao, Qiu-Yi Yin, Yan Jiang, Ting-Wang Jiang
{"title":"Triptolide-Loaded Solid Lipid Nanogel: Preparation and In-Vitro Evaluation","authors":"Chun-Feng Lu, Ye Dai, Yun Tao, Qiu-Yi Yin, Yan Jiang, Ting-Wang Jiang","doi":"10.1166/jbn.2024.3781","DOIUrl":"https://doi.org/10.1166/jbn.2024.3781","url":null,"abstract":"In order to topically deliver triptolide (TPL), we sought to develop and characterize solid lipid nano-particles based gel (SLNs-gel) before we investigated its inhibitory activity on HaCaT cells. Preparation of TPL-loaded SLNs (TPL-SLNs) was performed with a method involving melt-emulsion\u0000 ultra-sonication and solidification at low temperature. The determined characteristics of TPL-SLNs were particle size (PS), encapsulation efficiency (EE), zeta potential (ZP), microscopic mor phology and release of TPL In-Vitro. After TPL-SLNs have been formulated into gel, we used\u0000 the Franz diffusion cell method to evaluate the skin permeation and penetration characteristics of TPL-SLNs-gel on rat’s skin. Imaging results showed that particles of TPL-SLNs were homogeneous and well-dispersed. Meanwhile, the PS and ZP of TPL-SLNs were 89.21 ± 9.68 nm and −41.3\u0000 ± 6.23 mV, respectively, with EE being 89.3%. Also, we observed a significant improvement in pattern of In-Vitro TPL release from TPL-SLNs compared to free TPL. Furthermore, the cumulative penetration of TPL-SLNs-gel was higher (5.28 times) compared to free TPL. Besides, TPL-SLNs-gel\u0000 demonstrated substantial higher cytostatic activity on HaCaT cells comparable to both free TPL and TPL-SLNs. Altogether, it is evident that a delivery system like SLNs-gel can potentially increase the transdermal bioavailability of TPL for effective inhibition of proliferous HaCaT cells","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139890642","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}
Ji Ma, Yunna Ning, Ke Lu, Hui Wang, Ping Li, Lili Feng, Jianing Zhang, Linna Xie, Qiang He
{"title":"Hyper-Methylation of CpG Island in 5′ UTR of the HLA-G Gene Reduces Its Expression in Individuals with Immune Thrombocytopenia","authors":"Ji Ma, Yunna Ning, Ke Lu, Hui Wang, Ping Li, Lili Feng, Jianing Zhang, Linna Xie, Qiang He","doi":"10.1166/jbn.2024.3767","DOIUrl":"https://doi.org/10.1166/jbn.2024.3767","url":null,"abstract":"This study investigated the impact of DNA methylation in the 5′ untranslated region-CpG island (5′ UTR) of the HLA-G gene on soluble HLA-G (sHLA-G) levels in immune thrombocytopenia (ITP) patients, shedding light on sHLA-G’s regulatory mechanisms in ITP. Using a cohort\u0000 of 53 participants, including ITP patients, DNA methylation profiles in the HLA-G gene’s 5′ UTR were analyzed with Sequenom MassARRAY Methylation Analysis. sHLA-G levels were measured by enzyme-linked immunosorbent assay, and platelet antibodies were assessed using modified MAIPA.\u0000 Results showed increased DNA methylation at specific CpG sites (CpG3, CpG18, CpG19, and CpG20.21) in ITP patients. A negative correlation between DNA methylation and sHLA-G expression, particularly at CpG18, was found. Patients with Anti-GPIb/IX antibodies had higher CpG18 methylation. Age\u0000 and gender didn’t correlate significantly with methylation. This underscores 5′ UTR hypermethylation’s role in influencing circulating HLA-G levels, revealing insights into ITP development and potential therapeutic targets. By linking DNA methylation to sHLA-G expression,\u0000 this advances ITP understanding, suggesting new therapeutic strategies.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139817261","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}
Yang Xue, An Yan, Sara Amirpour Amraii, S. Goorani
{"title":"Determination of Cytotoxicity and Anti-Human Lung Cancer Properties of Silver Nanoparticles Green-Formulated by Foeniculum vulgare Extract","authors":"Yang Xue, An Yan, Sara Amirpour Amraii, S. Goorani","doi":"10.1166/jbn.2024.3775","DOIUrl":"https://doi.org/10.1166/jbn.2024.3775","url":null,"abstract":"Foeniculum vulgare is a plant with many therapeutic effects. In the current research, silver nanoparticles were synthesized by the Foeniculum vulgare extract. The properties of silver nanoparticles against lung cancer cell lines i.e., H69, COR-L47, DMS53, DMS79, NCI-H69/LX20,\u0000 SHP-77, NCI-H69/CPR, and NCI-H69/LX4 were evaluated. The green-formulated silver nanoparticles were characterized by various tests such as FE-SEM, EDX, FT-IR, and XRD. The FE-SEM findings prove spherical morphology for the AgNPs with the size of 19.34 to 47.93 nm. The IC50 of the silver nanoparticles\u0000 was 426, 547, 370, 377, 500, 384, 329, and 330 against H69, COR-L47, DMS53, DMS79, NCI-H69/LX20, SHP-77, NCI-H69/CPR, and NCI-H69/LX4, respectively. After doing the studies of clinical trial, the current nanoparticles may be used as an anti-lung cancer supplement in humans.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139821337","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}
{"title":"Study on the Mechanism and Protection of Salt-Sensitive Hypertensive Rats’ Myocardial Fibrosis by Regulating Striatin with Sacubatrovalsartan","authors":"Qingxian Tu, Qianhang Xia, Meihong Chen, Haiyan Zhou, Qianfeng Jiang, Wei Li","doi":"10.1166/jbn.2024.3766","DOIUrl":"https://doi.org/10.1166/jbn.2024.3766","url":null,"abstract":"This study aims to explore the relationship between STRN, TGF-β1, Caspase-3, PD-1 expression, and myocardial fibrosis in salt-sensitive hypertensive rats. It investigates the correlation between STRN expression and myocardial fibrosis, along with the protective effects of\u0000 Sacubitril/Valsartan (ARNI). Fifteen 18-week-old rats were divided into three groups: Control, high salt (SSH), and ARNI+SSH. Blood pressure was monitored weekly for 8 weeks. Echocardiography evaluated cardiac parameters, while H&E and Masson staining visualized myocardial morphology and\u0000 fibrosis. Immunohistochemistry measured protein expression of collagen-1, collagen-3, TGF-β1, PD-1, Caspase-3, and STRN. Western blot assessed STRN protein levels. High-salt diet increased fibrosis, collagen expression, TGF-β1, PD-1, Caspase-3, and reduced STRN expression\u0000 compared to Control (P < 0.05). ARNI treatment decreased fibrosis, collagen expression, TGF-β1, PD-1, Caspase-3 (P <0.05), and increased STRN expression compared to SSH (P <0.05). STRN expression correlated positively with myocardial fibrosis. ARNI\u0000 demonstrated potential in attenuating fibrosis by modulating STRN expression and suppressing apoptosis and inflammation in salt-sensitive hypertensive rats.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139821774","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}
Xue Li, Xiao Xiao, Lei Wang, Weichun Liang, Jun Ruan, Jianyi Ou
{"title":"A Sustained-Release SP-Sr-CaS/NBD Microsphere for Promoting Bone Repair and Inhibiting Inflammation for the Treatment of Osteomyelitis","authors":"Xue Li, Xiao Xiao, Lei Wang, Weichun Liang, Jun Ruan, Jianyi Ou","doi":"10.1166/jbn.2024.3762","DOIUrl":"https://doi.org/10.1166/jbn.2024.3762","url":null,"abstract":"In this study, we prepare sustained-release poly(lactide-co-glycolide) (PLGA)-based microspheres (SP), containing strontium (Sr), calcium sulfate (CaS), and NF-κB essential modifier-binding domain (NBD) peptide, namely SP-Sr-CaS/NBD, for the treatment of osteomyelitis.\u0000 Our results demonstrate that the SP-Sr-CaS/NBD group exhibited enhanced bone repair speed and infection clearance rate compared to other groups. Moreover, histological staining revealed more comprehensive bone structure restoration in the SP-Sr-CaS/NBD group. Furthermore, we assessed the levels\u0000 of bone growth factors and apoptosis factors in primary bone marrow mesenchymal stem cells and found that NBD effectively suppresses inflammation while Sr-CaS promotes bone healing by inhibiting cell apoptosis. Additionally, we conducted in vitro and in vivo toxicity evaluations\u0000 of the microspheres, which confirmed their potential as a competitive filling material for osteomyelitis. In conclusion, SP-Sr-CaS/NBD microspheres hold great promise as therapeutic scaffolds for clinical cases involving bone infections by reducing pain and treatment duration. This study provides\u0000 a new repair material for the treatment of osteomyelitis and promotes the development of repair material for osteomyelitis.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139826615","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}
{"title":"LncRNA AC003090.1 Mediates miR-203a-3p/FOXP1 Axis to Promote the Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells","authors":"Huafeng Zhuang, Xuedong Yao, Aifei Wang, Junjie Li, Miao Zheng, Yu Dong, Youjia Xu, Yizhong Li, Yongjun Lin","doi":"10.1166/jbn.2024.3744","DOIUrl":"https://doi.org/10.1166/jbn.2024.3744","url":null,"abstract":"We investigated the regulatory mechanism of lncRNA AC003090.1 in human bone marrow stem cells (hBMSCs). Tissues from patients with osteoporosis (OP) were collected for the detection and analysis of AC003090.1, miR-203a-3p, and FOXP1 expression by QPCR. The expression and activity of\u0000 alkaline phosphatase (ALP), an osteogenic marker, were detected using a modified Gomori calcium-cobalt assay and PNP colorimetric assay. Calcium deposition on the extracellular surface was demonstrated using alizarin red staining. Using the Oil Red O staining assay to detecte the Intracellular\u0000 lipid content. Dual luciferase reporting system verified the targeting relationship between AC003090.1 or FOXP1 and miR-203a-3p. qRT-PCR and Western blot were used to measure the expression level of β-catenin in hBMSCs after different intervention treatments. The expression levels\u0000 of AC003090.1 and FOXP1 were downregulated in Osteoporosis (OP), whereas those of miR-203a-3p were upregulated. An increase in AC003090.1 expression could enhance hBMSC osteogenic differentiation (OD) and reduce the adipogenic ability of hBMSCs. Furthermore, miR-203a-3p mimics or FOXP1 knock-down\u0000 reversed the effect of increased AC003090.1 expression on OD and adipogenic differentiation of hBMSCs. Dual luciferase reporter assays showed that AC003090.1 can sponge miR-203a-3p, which targets FOXP1 in hBMSCs. LncRNA AC003090.1 promotes OD of hBMSCs by regulating the miR-203a-3p/FOXP1 axis.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139828777","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}
{"title":"Effect of Polystyrene Targeting Nanoparticles on Lung Injury in Severe Acute Pancreatitis and NOX2/ROS/NF-κB Pathway","authors":"Changbo Liu, Liya Luo, Shuzhen Suo, Yongkang Song","doi":"10.1166/jbn.2024.3783","DOIUrl":"https://doi.org/10.1166/jbn.2024.3783","url":null,"abstract":"Relationship between polyethylene targeting nanoparticles and key components of the NOX2/ROS/NF-κB signaling pathway has not yet been fully clarified, and their regulatory role in lung injury in severe acute pancreatitis has not yet been confirmed. In this study, severe\u0000 acute pancreatitis lung injury cells were exposed to polyethylene targeting nanoparticles and MTT method was used to detect cell proliferation. Cell cycle and apoptosis rate were detected using flow cytometry and the expression of NOX2/ROS/NF-κB pathway was detected. The compound\u0000 polyethylene targeting nanoparticles inhibited proliferation of lung-damaged cells in severe acute pancreatitis dose-dependently (5, 10 and 20 μmol/L), induced G2/M phase arrest, and increased cell apoptosis. In addition, it reduced the expression of NOX2, ROS, and NF-κB,\u0000 indicating that NOX2/ROS/NF-κB pathway may be inhibited. Polystyrene targeting nanoparticles reduced the expression of IL-6, TNF-α, JAK, STAT, and IL-10. As a targeted drug delivery system, nano-drug-carrying systems help to dissolve drugs that are difficult to dissolve\u0000 in the drug solution and intervene in the corresponding tissues and cells in a targeted manner. The results of this study showed that polymer-targeted nano-drug systems could regulate the growth of lung-damaged cells in severe acute pancreatitis. Polyethylene targeting nanoparticles may be\u0000 effective in inhibiting inflammation in lung-damaged cells in severe acute pancreatitis via regulation of NOX2/ROS/NF-κB pathway.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139831547","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}
Qiang Sun, Jun Yao, Shuxun Wei, Xinxing Li, Weijun Wang
{"title":"The Value of Near-Infrared Multifunctional Nanoprobe Combined with Artificial Intelligence Microsensor Technology in Molecular Diagnosis for Gastric Cancer","authors":"Qiang Sun, Jun Yao, Shuxun Wei, Xinxing Li, Weijun Wang","doi":"10.1166/jbn.2024.3769","DOIUrl":"https://doi.org/10.1166/jbn.2024.3769","url":null,"abstract":"Since the symptoms of early gastric cancer patients are not obvious, the majority of new gastric cancer cases are progressive gastric cancer every year. In this paper, we applied nanomedicine technology to design and prepare multifunctional nanoparticles for the diagnosis and treatment\u0000 of gastric cancer. Through targeted imaging of gastric cancer, combined with phototherapy and the prepared nanoprobes are applied to the ectopic transplantation tumor model of gastric cancer. Meanwhile, a fluorescent microsensor based on graphene oxide and deoxyribonuclease is constructed\u0000 in order to realize the rapid detection of gastric cancer exosomes. The near-infrared multifunctional nanoprobe is combined with artificial intelligence microsensor technology and applied to the molecular diagnosis of gastric cancer. The results shows that the P-P-I-M+ laser irradiation group\u0000 has the highest fluorescence intensity and its average fluorescence intensity is 2.04 times higher than that of the P-P-I+ laser irradiation group. The relative cell viability of P-P-M+ laser irradiation group, P-P-I+ laser irradiation group and P-P-I-M+ laser irradiation group are 62.5%,\u0000 41.9% and 19.3%, respectively. Therefore, the method in this paper can reduce the non-specific toxicity to other organs as well as exert the effect of combining the diagnosis and treatment of gastric cancer.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139885179","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}
{"title":"The Impact of LncRNA Nuclear-Enriched Abundant Transcript 1-Mediated Regulation of ASK1 Expression via miR-20a on High Glucose-Induced Retinal Vascular Endothelial Cell Injury","authors":"Ling Zhao, Chunhua Cai, Congjian Yi, Chaobin Liu","doi":"10.1166/jbn.2024.3768","DOIUrl":"https://doi.org/10.1166/jbn.2024.3768","url":null,"abstract":"This study investigates how long non-coding RNA (LncRNA) NEAT1 influences high glucose-induced damage in human retinal vascular endothelial cells (hRECs). Different experimental groups were established, including normal, high glucose, LncRNA NEAT1 knockdown, and miR-20a inhibition.\u0000 Assessments were conducted for molecular and functional changes. In high glucose conditions, NEAT1 expression increased while miR-20a expression decreased in hRECs. Silencing NEAT1 reduced its levels and increased miR-20a expression. Consequently, reactive oxygen species (ROS), MDA, 4-HNE,\u0000 IL-1β, TNF-α, ICAM-1, ASK1, VEGF, and p-p38 MAPK/p38 MAPK ratio decreased. This led to diminished cell proliferation, migration, and tube formation in hRECs. The impact of NEAT1 silencing was partially reversed by miR-20a inhibition, suggesting NEAT1′s regulatory\u0000 role via miR-20a. NEAT1 suppressed miR-20a and ASK1 protein levels. Additionally, LncRNA NEAT1 sequestered miR-20a, contributing to ASK1 downregulation. This process also suppressed p38 MAPK activation, further inhibiting hREC functions. In summary, NEAT1 modulated high glucose-induced hREC\u0000 injury by downregulating miR-20a and subsequently impacting ASK1 and p38 MAPK pathways, thereby impairing cell functions. This study provides insights into potential therapeutic targets for diabetic retinopathy.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139887144","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}
{"title":"Antibacterial Effect of Nanostructured Cuprous Gold and Silver Oxide Composite Nanoshell on Keratitis","authors":"Chen Wang, Yang Liu, Mingchang Zhang","doi":"10.1166/jbn.2024.3764","DOIUrl":"https://doi.org/10.1166/jbn.2024.3764","url":null,"abstract":"We developed novel inorganic nanomaterials to combat drug-resistant bacterial infections in keratitis. These infections cause rapid severe corneal ulcers. Traditional antibiotics face challenges due to bacterial resistance. We investigated new therapies by designing nanomaterials. In\u0000 an animal model of diabetic keratitis, we studied the materials’ antibacterial properties and mechanisms. In vitro, nanomaterials displayed strong antibacterial effects, confirmed by quantitative analysis. In vivo, using thermal imaging, wound closure monitoring, clinical\u0000 scores, and histopathology, we demonstrated nanomaterials’ efficacy against infections. Toxicity evaluations, including weight monitoring, hemolysis, biochemical, hematological analyses, and organ histology, revealed no adverse effects on the body or organs. Confocal microscopy showed\u0000 effective bacterial eradication using nanomaterials combined with near-infrared laser treatment. Minimal impact on red blood cells was observed at therapeutic concentrations. Nanomaterials, particularly gold-silver-cuprous oxide composite nanoshells, demonstrated potent resistance against\u0000 drug-resistant infections. Photothermal treatment using nanomaterials and near-infrared laser showed promise without harming normal tissues, blood, or organs. Our findings offer a potential clinical solution for keratitis treatment.","PeriodicalId":15260,"journal":{"name":"Journal of biomedical nanotechnology","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139874015","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}