Stem Cell Reviews and Reports最新文献

{"title":"Electron Beam Irradiation Induces Senescence Alterations in Human Adipose-Derived Stem Cells.","authors":"Denis Baranovskii, Svetlana Lyamina, Anastas Kisel, Sergey Kalish, Ekaterina Kozhevnikova, Tatiana Ivanova, Elena Isaeva, Vadim Govorun, Ilya D Klabukov","doi":"10.1007/s12015-025-10867-y","DOIUrl":"10.1007/s12015-025-10867-y","url":null,"abstract":"","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"1138-1140"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143674521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Self-assembly and 3D Bioprinting of Neurospheres and Evaluation of Caffeine and Photobiomodulation Effects in an Alzheimer's Disease In Vitro Model.","authors":"Geisa Rodrigues Salles, Alessandro E C Granato, Fernanda Tibolla Viero, Cristina Pacheco-Soares, Sérgio T Ferreira, Marimelia Porcionatto, Henning Ulrich","doi":"10.1007/s12015-025-10850-7","DOIUrl":"10.1007/s12015-025-10850-7","url":null,"abstract":"<p><p>Several in vitro models of Alzheimer's disease (AD) rely on 2D cell culture, and, more recently, 3D cultures represented by free-floating neurospheres have been used as models for the disease. The advantage of 3D over 2D cell culture is that cell-extracellular matrix and cell-cell interactions can be assessed, better representing the molecular and cellular hallmarks of the disease. In the current study, we developed two complementary 3D neurosphere models using SH-SY5Y human neuroblastoma cells to investigate AD pathology and evaluate potential therapies. First, self-assembled neurospheres were exposed to hydrogen peroxide (H₂O₂) and amyloid-beta oligomers (AβOs), inducing AD-like features such as increased production of reactive oxygen species (ROS), amyloid aggregation, and apoptosis. Treatment with caffeine or photobiomodulation (PBM) using LED irradiation significantly reduced Aβ_1-42accumulation, ROS generation, and decreased apoptosis markers. Second, 3D bioprinting of SH-SY5Y cells resulted in neurospheres with enhanced cellular organization and differentiation. These findings emphasize the advantages of 3D models for studying neurodegeneration and evaluating therapeutic strategies, bridging the gap between traditional 2D cultures and complex in vitro systems.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"988-1000"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143812309","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of the Safety and Regenerative Potential of Human Mesenchymal Stem Cells and Their Extracellular Vesicles in a Transgenic Pig Model of Cartilage-Bone Injury In Vivo - Preclinical Study.","authors":"Anna Łabędź-Masłowska, Jarosław Wieczorek, Maciej Mierzwiński, Małgorzata Sekuła-Stryjewska, Sylwia Noga, Jolanta Rajca, Piotr Duda, Katarzyna Milian-Ciesielska, Elżbieta Karnas, Katarzyna Kmiotek-Caller, Agnieszka Szkaradek, Zbigniew Madeja, Krzysztof Ficek, Jacek Jura, Ewa Zuba-Surma","doi":"10.1007/s12015-025-10853-4","DOIUrl":"10.1007/s12015-025-10853-4","url":null,"abstract":"<p><p>Osteoarthritis (OA) is a degenerative joint condition leading to disability. The lack of effective treatment for OA creates a need for the development of new therapeutic approaches that may rely on stem cells including mesenchymal stem/stromal cells (MSCs) and their derivatives such as extracellular vesicles (EVs). The objective of this study was to evaluate the impact of MSCs derived from adipose tissue (AT-MSCs) and umbilical cord (UC-MSCs) and their EVs on cartilage-bone injury in vivo, to identify the specimen with the highest regenerative potential for further clinical applications in patients with OA. Humanized pigs underwent cartilage-bone injuries followed by intraarticular administration of products containing AT-MSCs, UC-MSCs, AT-MSC-EVs or UC-MSC-EVs mixed with hyaluronic acid (HA) or HA alone (for comparison). After 6-m follow-up, almost-fully-healed cartilage-bone defects were observed in the AT-MSC- and UC-MSC-treated pigs, and the defects were filled primarily with hyaline cartilage. In AT-MSC-EV- and UC-MSC-EV-treated pigs, a partial cartilage-bone tissue repair was observed, and the defects were filled primarily with fibrocartilage. The control pigs demonstrated limited regeneration capacity. The microcomputed tomography parameters of the subchondral bone indicated the ongoing progression of OA in controls, whereas in the MSC- and MSC-EV-treated pigs, the parameters indicated the cessation of OA progression. Moreover, no serious side effects were observed after the administration of products containing MSCs or MSC-EVs. The results indicate the safety and regenerative activity of MSCs on injured tissues, which favors not only the healing and improvement of bone structure but also the formation of hyaline cartilage. Superior tissue repair was observed after the administration of products containing AT-MSCs. The treatment of OA with MSC-EVs needs further standardization.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"1075-1095"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12102096/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144086737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Compatibility of Hepatocyte Growth Factor, Adipose-Derived Mesenchymal Stem Cells, and Dermal Fillers for the Treatment of Vocal Fold Scar.","authors":"Nabanita Halder, Ralph Perkerson, Takahisa Kanekiyo, Amy L Rutt","doi":"10.1007/s12015-025-10862-3","DOIUrl":"10.1007/s12015-025-10862-3","url":null,"abstract":"<p><p>Vocal fold (VF) scar and sulcus cause severe vocal problems, however optimal treatment methods are not well established. We tested the viability of mesenchymal stem cell (MSC) therapy in combination with hepatocyte growth factor (HGF) in a hyaluronic acid-based (HA) hydrogel. There was a significant increase in the metabolic output of adipose- derived mesenchymal stem cells (AD-MSC) grown in the presence of HGF up to 1 ug/mL when compared to control wells. Metabolic activity of AD-MSC in the presence of 2 ug/mL HGF was similar to controls. No significant differences were detected between any of the groups when compared to control wells. The results from these experiments provide evidence for the compatibility of the tested products; recombinant human HGF, human AD-MSC, and HA-based dermal fillers. These pre-clinical results support the ongoing research of this combination therapy in treating VF scar.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"1131-1133"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143524467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Complosome Regulates Hematopoiesis at the Mitochondria Level.","authors":"Adrian Konopko, Agnieszka Łukomska, Janina Ratajczak, Magdalena Kucia, Mariusz Z Ratajczak","doi":"10.1007/s12015-025-10856-1","DOIUrl":"10.1007/s12015-025-10856-1","url":null,"abstract":"<p><p>The intracellular complement network, known as the complosome, regulates lymphocyte biology, which is well established. Recently, however, we demonstrated that the complosome is also expressed in hematopoietic stem/progenitor cells (HSPCs) in addition to lymphocytes. In our previous work, murine lineage-negative (Lin^-) bone marrow (BM) mononuclear cells (BMMNC) from mice lacking the intracellular C3 and C5 complosome proteins displayed different responses to stress. Specifically, while C3-KO cells were more sensitive to oxidative stress, C5-KO cells showed greater resistance. To explore this intriguing observation at the metabolic level, we evaluated anaerobic and aerobic glycolysis, along with mitochondrial function, in Lin^- BMMNC purified from C3-KO, C5-KO, and C5aR1-KO mice. We found that cells from complosome-deficient animals under steady-state conditions exhibited elevated lactate production and enhanced lactate dehydrogenase (LDH) release, indicating their reliance on anaerobic glycolysis. Interestingly, the uptake of a glucose fluorescent analog (2-NBDG) increased in C3-KO cells but decreased in C5-KO and C5aR1-KO cells compared to wild-type (WT) mice. Meanwhile, total ATP production in C3-KO cells, unlike that of C5 and C5aR1 mice, was reduced under steady-state conditions and did not change significantly after exposure to the mitochondrial-damaging agent hydrogen peroxide (H₂O₂). This suggests a greater dependence on anaerobic glycolysis in C3-KO cells than in C5-KO and C5aR1-KO cells. Finally, we assessed the integrity of mitochondrial membranes in the studied cells using MitoTracker green and deep red assays. Compared to WT cells, we observed that mitochondria from complosome mutant Lin-BMMNC accumulated fewer MitoTracker probes, indicating the presence of mitochondrial defects in these cells.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"1001-1012"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12102138/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143573894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advances in the Regulation of Hematopoietic Homeostasis by Programmed Cell Death Under Radiation Conditions.","authors":"Manling Shu, Jinfu Zhang, Haocong Huang, Yuxin Chen, Yubing Shi, Huihong Zeng, Lijian Shao","doi":"10.1007/s12015-025-10863-2","DOIUrl":"10.1007/s12015-025-10863-2","url":null,"abstract":"<p><p>The application of nuclear energy and the frequent occurrence of nuclear contamination have made radiation safety a major challenge to global public health. As a radiation-sensitive target organ, bone marrow is susceptible to both acute and chronic damage effects of ionizing radiation on the hematopoietic system. Researchers have demonstrated that radiation disrupts hematopoietic homeostasis through direct damage to hematopoietic stem cells, which inhibits hematopoietic regeneration indirectly through damage to hematopoietic progenitor cells and their downstream cell populations. However, the multi-target regulatory mechanism of radiation perturbation of hematopoietic homeostasis remains to be systematically elucidated. Recent studies have revealed that, in addition to the classical apoptotic pathway, non-apoptotic programmed cell death modes (e.g. pyroptosis, necroptosis, and ferroptosis) may be involved in the regulation of radiation-induced hematopoietic injury. A systematic review of the roles of the aforementioned programmed death pathways was presented in radiation-damaged hematopoietic cells, with a view to providing a scientific basis for targeted intervention in radiation-induced myelosuppression.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"935-952"},"PeriodicalIF":4.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143582340","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fibroblast-Derived Human iPSC Exhibits Superior Haematopoietic Potential over Human ESC during Haematopoietic Differentiation.","authors":"Yee-Ching Lim, Soon-Keng Cheong, Pooi-Pooi Leong","doi":"10.1007/s12015-025-10855-2","DOIUrl":"10.1007/s12015-025-10855-2","url":null,"abstract":"<p><p>Haematopoietic stem cells (HSC) and macrophages hold promise for cell-based therapy. Induced pluripotent stem cells (iPSC) offer an alternative to human embryonic stem cells (hESC) for generating haematopoietic cells in vitro, sidestepping ethical concerns. However, precise comparisons of the developmental process and productivity between iPSC and hESC during haematopoietic differentiation are limited, and producing sufficient HSC for clinical use remains challenging. We introduce a refined, simplified protocol that is xeno-, serum-, and feeder-free for differentiating fibroblast-derived human iPSC (NHDF-iPSC) and the hESC-H9 clone (H9-ESC) using the STEMdiff™ Hematopoietic kit, with differentiation extended by in-house cytokine addition. We demonstrate that NHDF-iPSC recapitulate the haematopoietic differentiation of H9-ESC, forming CD31⁺CD144⁺CD34⁺ haemogenic endothelia (HE) as intermediates, and producing CD34⁺CD43⁺CD45^+/- haematopoietic stem and progenitor cells (HSPC). This protocol facilitates the production of CD34⁺ HSPC over an extended period and enhances the yield of HSC from NHDF-iPSC-derived HE three-fold. Interestingly, our results demonstrated that NHDF-iPSC outperformed H9-ESC by exhibiting superior differentiation capabilities, resulting in a higher abundance of HE and greater haematopoietic cell output (e.g., HSPC and HSC) upon cytokine stimulation. This phenomenon is presumably due to the higher expression of RUNX1 in NHDF-iPSC-derived HE (three-fold) as observed in our study, which may lead to a more productive endothelial-to-haematopoietic transition process and potentially facilitate the efficient production of haematopoietic cells. These CD34⁺ haematopoietic cells mature into 25F9⁺CD45⁺ macrophages, which exhibit comparable functions to those derived from hESC. Together, our results underscore the potential of iPSCs as a sustainable source for deriving HSC and macrophages for cell-based therapies.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"1034-1047"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504285","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Current Applications of Human Pluripotent Stem Cells in Neuroscience Research and Cell Transplantation Therapy for Neurological Disorders.","authors":"Isha Verma, Polani B Seshagiri","doi":"10.1007/s12015-025-10851-6","DOIUrl":"10.1007/s12015-025-10851-6","url":null,"abstract":"<p><p>Many neurological diseases involving tissue damage cannot be treated with drug-based approaches, and the inaccessibility of human brain samples further hampers the study of these diseases. Human pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), provide an excellent model for studying neural development and function. PSCs can be differentiated into various neural cell types, providing a renewal source of functional human brain cells. Therefore, PSC-derived neural cells are increasingly used for multiple applications, including neurodevelopmental and neurotoxicological studies, neurological disease modeling, drug screening, and regenerative medicine. In addition, the neural cells generated from patient iPSCs can be used to study patient-specific disease signatures and progression. With the recent advances in genome editing technologies, it is possible to remove the disease-related mutations in the patient iPSCs to generate corrected iPSCs. The corrected iPSCs can differentiate into neural cells with normal physiological functions, which can be used for autologous transplantation. This review highlights the current progress in using PSCs to understand the fundamental principles of human neurodevelopment and dissect the molecular mechanisms of neurological diseases. This knowledge can be applied to develop better drugs and explore cell therapy options. We also discuss the basic requirements for developing cell transplantation therapies for neurological disorders and the current status of the ongoing clinical trials.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"964-987"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143789030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"G-CSF-Induced Emergency Granulopoiesis Modulates Neutrophil Effector Function in Mice.","authors":"Jonah K Stephan, Taylor Knerr, Collin K Wells, Zhen Gu, Sidney Johnson, Tyler K Jobe, William S Isaacs, Bradford G Hill, Marcin Wysoczynski","doi":"10.1007/s12015-025-10885-w","DOIUrl":"10.1007/s12015-025-10885-w","url":null,"abstract":"<p><p>Neutrophils function as first responders of the immune system by deploying cytotoxic armaments and orchestrating local inflammation. Their functionality is programmed during daily production in the bone marrow through granulopoiesis. During severe inflammation, increased neutrophil demand is met through activation of emergency granulopoiesis. The effect of emergency granulopoiesis on neutrophil functionality remains cryptic. In the present study, we assessed neutrophil function in mice injected with G-CSF (100 µg/kg/d for 3 days) to activate emergency granulopoiesis. We found that emergency granulopoiesis neutrophils exhibit impaired ROS production (n = 6, P = 0.003) and NETosis (n = 5, P < 0.01), but increase neutrophil elastase secretion (n = 9, P < 0.0001) and LPS-induced Tnfa, Il1b, Il1a, Il12a, and Ccl2 expression (n = 13, P < 0.01). To test the impact of emergency granulopoiesis neutrophils on the inflammatory response in vivo, we pre-treated mice with G-CSF and challenged them with zymosan to induce peritonitis. At 4 h post-zymosan injection, peritoneal neutrophils from G-CSF treated mice exhibit increased expression of Ccl2 (n = 3, P < 0.05). Subsequently, we observed enhanced peritoneal macrophage accumulation at 48 h post-zymosan administration in G-CSF-treated mice (n = 5, P < 0.05). These data indicate that emergency granulopoiesis programs neutrophils to have an enhanced immunomodulatory function that orchestrates the subsequent macrophage response in local tissue inflammation.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"1113-1126"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12167617/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143997850","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Human Umbilical Cord Mesenchymal Stem Cell-Derived Exosomes Boost Functional Performance in an Animal Model of Multiple Sclerosis Through Recruiting Oligodendrocytes and Attenuating Gliosis.","authors":"S Mohammadhadi Mirab, Ameneh Omidi, Masoud Soleimani, Mina Soufi-Zomorrod, Zahra Fekrirad","doi":"10.1007/s12015-025-10858-z","DOIUrl":"10.1007/s12015-025-10858-z","url":null,"abstract":"<p><strong>Background: </strong>Multiple sclerosis (MS) is a chronic neuroinflammatory and demyelinating condition that causes movement problems due to a probable failure to differentiate oligodendrocyte precursor cells into myelinating oligodendrocytes.</p><p><strong>Aims: </strong>This work aims to evaluate the effect of exosomes derived from human umbilical cord mesenchymal stem cells on functional recovery in an animal model of multiple sclerosis.</p><p><strong>Methods: </strong>The adult male C57BL/6J mice were randomly divided into three groups. The control mice were provided with a standard diet (CONT group). The chronically demyelinated (CPZ) group was fed a 0.2% cuprizone diet for 12 weeks. In the third group, mice received exosomes weekly for three weeks following chronic demyelination (Exo/CPZ group). The mice in the groups were tested weekly for mobility using a beam walking test (BWT), and their corpus callosum was studied histologically, immunohistochemically, and molecularly.</p><p><strong>Results: </strong>According to BWT results, hucMSC-Exos enhanced motor function in mice following cuprizone intoxication. Histological staining revealed a substantial increase in remyelination in the corpus callosum. Moreover, immunohistochemical tests revealed that hucMSC-Exos lowered the level of demyelination and improved glial response by lowering the number of microglia and astrocytes. In addition, hucMSC-Exos also upregulated the expression of genes associated with the oligodendrocyte lineage.</p><p><strong>Conclusions: </strong>hucMSC-Exos administration in the animal model of chronic MS leads to improved movement function, likely due to increased remyelination, modulation of inflammatory processes, and increased expression of genes related to the oligodendrocyte lineage. Therefore, using hucMSC-Exos seems to be a promising treatment strategy for demyelinating illnesses such as MS.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":"1048-1061"},"PeriodicalIF":4.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143573863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}