Stem Cell Reviews and Reports最新文献

{"title":"From Design to Single-Cell Cloning: A Complete RNP-Based CRISPR-Cas9 Protocol for Precision Gene Correction in Human iPSCs.","authors":"Giacomo Roman, Knut H Lauritzen, Barbora Smolkova, Runar Almaas, Per Morten Sandset, Benedicte Stavik, Gareth J Sullivan, Maria E Chollet","doi":"10.1007/s12015-026-11221-6","DOIUrl":"10.1007/s12015-026-11221-6","url":null,"abstract":"<p><p>Precise genome editing of induced pluripotent stem cells (iPSCs) using clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9 (CRISPR-Cas9) has opened unprecedented avenues for advancements in regenerative medicine and disease modelling. However, the establishment of isogenic single-cell-derived iPSC populations, particularly upon CRISPR-Cas9 gene editing modifications, is one of the major challenges still associated with these advanced methodologies relying on low-rate editing events and requiring defined clonogenicity. In response, we have developed a systematic, comprehensive and efficient workflow combining generation, genotyping and expansion of high-quality monoclonal iPSC lines following CRISPR-Cas9 genome editing. In particular, the protocol incorporates optimized single-cell cloning procedures for two commercially available dispensing platforms, one based on microfluidic imaging and the other on impedance technology, together with rapid droplet digital PCR (ddPCR)-based screening of non-homologous end joining (NHEJ) and homology-directed repair (HDR) outcomes. By combining gentle single-cell handling with advanced genotyping methodologies, the protocol enables efficient early assessment of editing outcomes before commitment to labour-intensive clonal derivation, thereby accelerating project timelines, minimising cell stress and loss, preserving genetic fidelity and supporting scalability. Coupled with precisely defined culture conditions tailored for post-seeding recovery, these approaches aim to improve iPSC viability and clonal outgrowth, achieving at least 60% in 96-well plate format within 10 days. Importantly, the protocol goes beyond step-by-step experimental instructions by providing comprehensive design strategies, decision-making criteria, and practical advice for avoiding and addressing the common pitfalls and unintended consequences of these advanced methods. Collectively, this integrated end-to-end approach provides a robust and high-throughput framework for the reliable production of monoclonal genome-edited iPSCs, thereby advancing translational research and the development of iPSC-based regenerative therapies and disease models.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148865997","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":"Mechanistic Study of m5C Modification in Stem Cell Differentiation and Disease Regulation.","authors":"Xiangrong Zhang, Jianqiao Li, Zhifeng Zhang, Xueqian Chen, Jiankai Yang, Wei Liu","doi":"10.1007/s12015-026-11222-5","DOIUrl":"https://doi.org/10.1007/s12015-026-11222-5","url":null,"abstract":"<p><p>m5C (5-methylcytosine) modification is a significant epigenetic modification in RNA that has garnered substantial attention in recent years in stem cell biology and disease research. The dynamic regulation of m5C involves methyltransferases, demethylases, and specific binding proteins, which collectively influence RNA stability, translation efficiency, and cellular function. Especially in stem cell differentiation, m5C modification not only affects stem cell self-renewal and differentiation fate but is also closely associated with the onset and progression of various diseases. This review explores the mechanisms of m5C modification in stem cell differentiation and related diseases, highlighting its emerging roles in gene expression regulation and disease pathogenesis. CLINICAL TRIAL NUMBER: Not applicable.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148851270","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":"\"Don't Force the Stem Cell for Hair Regeneration: Rather Electrically and Functionally Prime the Fibroblast.\"","authors":"Mamata Mishra, Rinky Kapoor, Khushbu Kumari, Debraj Shome","doi":"10.1007/s12015-026-11225-2","DOIUrl":"https://doi.org/10.1007/s12015-026-11225-2","url":null,"abstract":"","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148857603","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":"Maternal and Neonatal Factors Impacting the Volume and Cellular Composition of Umbilical Cord Blood.","authors":"Yu Li, Jia Liu, Ming Li, Chunyu He, Bin Guo, Ye Yang, Rongzhen Jiang","doi":"10.1007/s12015-026-11205-6","DOIUrl":"https://doi.org/10.1007/s12015-026-11205-6","url":null,"abstract":"<p><strong>Background: </strong>Umbilical cord blood (UCB) is a well-established source of hematopoietic stem cells (HSCs) for transplantation, with over 30 years of clinical experience confirming its safety and efficacy. However, the limited cell count contained for a single UCB unit remains a major constraint on its application in adult recipients, and donor eligibility criteria vary greatly among different cord blood banks.</p><p><strong>Objective: </strong>This study aimed to identify key maternal, neonatal, and perinatal factors that influence the volume and quality of collected UCB to help overcome this limitation.</p><p><strong>Methods: </strong>We conducted a retrospective analysis of 229 UCB units voluntarily donated by pregnant women at our hospital. Correlations between UCB parameters and a range of clinical characteristics were evaluated.</p><p><strong>Results: </strong>Maternal body mass index (BMI), umbilical cord length, and UCB oxygen partial pressure (pO₂) were independently associated with UCB volume and cellular parameters. Furthermore, umbilical cord length and maternal platelet-to-lymphocyte ratio (PLR) showed associations with TNC count in univariate analysis, although no independent factors were identified after multivariable adjustment. The CD34⁺/TNC ratio was independently associated with umbilical cord length, gestational age, maternal total cholesterol level, placental weight, and UCB pH value. Finally, umbilical cord length was independently associated with the total CD34⁺ cell count.</p><p><strong>Conclusion: </strong>These findings provide preliminary evidence that selected maternal, neonatal, and perinatal characteristics are associated with UCB volume and cellular parameters. Further multicenter studies incorporating functional stem-cell assessments and transplantation outcomes are required before changes to donor eligibility or UCB unit-selection criteria can be considered.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148814076","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":"Adipose-Derived Stem Cells in Ophthalmic Regenerative Medicine: From Cell Replacement to Precision Engineering.","authors":"Bei Li, Le Ling, Ruixi Liu, Xin Zhang, Yifan Kang, Jinlin Guo","doi":"10.1007/s12015-026-11220-7","DOIUrl":"https://doi.org/10.1007/s12015-026-11220-7","url":null,"abstract":"<p><p>Adipose‑derived stem cells (ASCs) have emerged as a promising cell source in ophthalmic regenerative medicine, attributed to their accessibility, differentiation potential, and paracrine functions. Despite compelling preclinical evidence, clinical outcomes remain unsatisfactory. Departing from the conventional disease‑oriented categorization, this review adopts the cognitive evolution of ASC therapeutic mechanisms as its organizational thread, synthesizing evidence from 158 studies across six ophthalmic subspecialties. Four sequential paradigms are identified: (1) multilineage differentiation and cell replacement; (2) paracrine signaling and microenvironmental modulation; (3) extracellular vesicles and cell‑free therapy; and (4) engineering modifications and precision delivery. Cross‑disciplinary synthesis reveals a fundamental contradiction: the inherent capacity of living ASCs to sense and respond to local microenvironments directly conflicts with the pharmacological requirements for standardization and predictability. Five major translational bottlenecks are distilled, namely, the absence of functional potency standards, the rodent‑to‑human translational gap, insufficient therapeutic window studies, low‑level clinical evidence outside oculoplastics, and regulatory ambiguity. A corresponding roadmap is proposed, centered on functional potency evaluation, engineered cell‑free products, and an ophthalmic‑specific long‑term safety framework. This review provides a unified theoretical framework for contextualizing the prospects of ASC‑based therapies and offers strategic guidance to facilitate their clinical translation.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798017","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":"Inner Ear Organoids: Recent Progress and Challenges.","authors":"Yingxinyu Li, Cairong He, Jiayan Liu, Haiqin Zhang, Hongxia Zhang, Zheyu He, Mengyang He, Meiling Chen, Feiyan Zhou, Jie Ding","doi":"10.1007/s12015-026-11219-0","DOIUrl":"https://doi.org/10.1007/s12015-026-11219-0","url":null,"abstract":"<p><p>Hearing loss is a prevalent sensory disorder primarily caused by the irreversible loss of cochlear hair cells and the limited regenerative capacity of the mammalian inner ear. The structural complexity and inaccessibility of inner ear tissues have long hindered mechanistic studies and therapeutic development. In recent years, inner ear organoids have emerged as a robust in vitro platform that recapitulates key aspects of inner ear development, cellular composition, and functional organization. Derived from pluripotent stem cells or tissue-specific progenitors, inner ear organoids exhibit intrinsic self-organizing capabilities under the spatiotemporal regulation of developmental signaling pathways, including TGF-β, BMP, FGF, and Wnt. These systems enable the generation of hair cell-like cells, supporting cells, and neuronal components, thereby providing a powerful tool for investigating developmental processes and disease mechanisms. Notably, patient-specific induced pluripotent stem cell-derived organoids facilitate modeling of hereditary hearing disorders and enable personalized therapeutic screening. Beyond basic research, inner ear organoids hold significant promise for drug discovery, ototoxicity assessment, and gene therapy validation. However, current models still face several limitations, including incomplete cellular heterogeneity, insufficient functional maturation, lack of vascularization and immune components, and variability across culture systems. Emerging bioengineering approaches, such as organ-on-a-chip platforms, synthetic biomaterials, and 3D bioprinting, are expected to enhance the physiological relevance and translational potential of these models. In this review, we summarize recent advances in the generation of inner ear organoids, discuss their applications in basic and translational research, and highlight current challenges and future directions toward clinical translation.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148797964","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":"Dysfunction of the Neurovascular Unit in Diabetic Retinopathy: Advances from Molecular Mechanisms to Targeted Interventions.","authors":"Chunning Zhao, Leilei Qin, Jing Zhang, Xiu Wang","doi":"10.1007/s12015-026-11213-6","DOIUrl":"https://doi.org/10.1007/s12015-026-11213-6","url":null,"abstract":"<p><p>Diabetic retinopathy (DR) is a leading cause of vision loss and is now increasingly understood as a neurovascular disease rather than a purely microvascular complication of diabetes. The retinal neurovascular unit (NVU), composed of neurons, Müller cells, microglia, astrocytes, endothelial cells, pericytes, and extracellular matrix components, integrates neural activity, vascular perfusion, barrier integrity, and inflammatory control. In diabetes, NVU injury develops through a hierarchical and self-amplifying process. Upstream metabolic and systemic triggers, including hyperglycemia, advanced glycation end products (AGEs), homocysteine and lipid imbalance, and NADPH oxidase-derived reactive oxygen species, initiate cellular stress. These triggers converge on mitochondrial dysfunction, impaired autophagy and mitophagy, endoplasmic reticulum stress, and mitochondrial DNA release, which subsequently activate innate immune pathways. Neuroinflammatory amplification driven by microglial activation, Müller cell gliosis, NLRP3 inflammasome signaling, pyroptosis, and cytokine feedback further links neuronal and vascular injury. The final common pathway is integrated NVU breakdown, characterized by retinal neurodegeneration, pericyte loss, endothelial dysfunction, blood-retinal barrier disruption, vascular leakage, and VEGF-driven neovascularization. Regulatory RNA networks, including lncRNAs, miRNAs, circRNAs, piRNAs, and tRNA-derived fragments, act as cross-cutting epigenetic regulators of these processes. This review integrates current evidence within an NVU-centered systems biology framework and aligns therapeutic strategies with the disease hierarchy, distinguishing established clinical treatments from preclinical and emerging interventions.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148797961","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":"A distinct Frequency of Endometrial Stem/Progenitor Cells (EnSCs) in the Ectopic and Eutopic Endometrium of Women with Endometriosis.","authors":"Mousumi Bal, Vivek Salunke, Shinjini Pande, Geetanjali Sachdeva, Dhanjit K Das","doi":"10.1007/s12015-026-11216-3","DOIUrl":"https://doi.org/10.1007/s12015-026-11216-3","url":null,"abstract":"<p><p>Endometriosis is a chronic estrogen-dependent disorder characterised by the growth of endometrial-like tissues outside the uterus. Recent data suggest a potential role for endometrial stem/progenitor cells in the development of endometriosis. However, evidence on the frequency of various stem cell populations in ectopic lesions and their paired eutopic counterparts remains limited. Hence, this study assessed the frequencies of endometrial stem cells and their characteristics in eutopic and ectopic endometria of women with endometriosis, and also probed whether these attributes differ in women without endometriosis. We analysed the frequencies of endometrial stem cells and their functional properties, isolated from ectopic lesions and paired eutopic endometrium, in women with endometriosis (n = 72) and women without endometriosis (n = 24). Comparative analysis revealed that the eutopic endometrium had a significantly increased frequency of SUSD2+ eMSCs in women with endometriosis than in those without the disease. Immunophenotypic characterisation of SUSD2+ eMSCs using classical MSC markers (CD73, CD90, CD105, and HLA-ABC) revealed that they retained more than 80% positivity in the eutopic endometrium, whereas variable expression of these markers was observed among ectopic lesions. On functional characterisation, the wound healing assay revealed that eMSCs from deep infiltrating endometriosis have increased migratory capacity compared to control and paired eutopic endometrium. Moreover, a preponderance of eEPCs (N-cadherin+ SSEA-1+ and N-cadherin- SSEA-1+) was observed in the eutopic endometrium of endometriosis. In contrast, a significantly increased frequency of Side Population was observed in ectopic lesions compared to eutopic endometrium. Besides Side Population, ovarian endometrioma exhibited a significantly increased frequency of pericyte eMSCs co-expressing CD146 and CD140b. These findings clearly demonstrated a distinct frequency of endometrial stem/progenitor cells in endometriosis. Collectively, this study underscores the predominance of endometrial stem cells and highlights altered immunophenotypic characteristics and enhanced migration capabilities of ectopic eMSCs, possibly driven by unique cellular microenvironments.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148797970","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":"Dental Pulp Regeneration: A Comprehensive Review of Stem Cells, Biomaterials, and Bioactive Cues.","authors":"Yeshi Pei, Cheng Dong","doi":"10.1007/s12015-026-11214-5","DOIUrl":"https://doi.org/10.1007/s12015-026-11214-5","url":null,"abstract":"<p><p>Dental pulp regeneration (DPR) is an important research direction in regenerative endodontics that seeks to restore pulp vitality, sensation, immune defense, and the pulp-dentin architecture rather than merely retain a non-vital tooth. This review provides a mechanistically coupled and clinically focused synthesis of three interacting elements: stem/progenitor cells, biomaterials, and bioactive signals. Dental and non-dental cell sources are compared with emphasis on their angiogenic, neurogenic, odontogenic, and immunomodulatory characteristics and on the donor- and culture-dependent variability that affects product potency. Rather than presenting scaffolds only by material class, the review offers a structured critical correlation of stiffness, viscoelasticity, porosity, degradation, and bioactivity with cell fate, extracellular matrix organization, vascularization, and the risk of ectopic mineralization. Regulated growth-factor presentation, peptide-based signals, gene-activated constructs, extracellular vesicles, immune-instructive materials, and mechanobiological approaches are evaluated according to the highest experimental model in which they have been demonstrated. A stratified research framework is proposed to examine how age, inflammatory phenotype, systemic disease, apical anatomy, and regional heterogeneity of the pulp microenvironment may influence regenerative responses. The supporting evidence remains predominantly in vitro or preclinical. Cell-free regenerative endodontic procedures are clinically accepted for selected immature necrotic teeth with open apices, but available human histology most often demonstrates repair rather than recreation of a native pulp-dentin complex. Human cell transplantation studies remain small and early-phase, and advanced gene-, vesicle-, biofabrication-, and computational strategies have not yet demonstrated clinical efficacy. The framework is therefore presented as an evidence-graded research roadmap rather than a description of established personalized therapy.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798005","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":"Extracellular Vesicles Derived from Resveratrol-Treated BMSCs Promote the Repair of Bone Defects in Osteoporotic Rats.","authors":"Yan Lv, Yongjun Du, Zhi Peng, Chengjie Ma, Zhenggang Li, Rui Xu, Jianhong Hou, Sheng Lu","doi":"10.1007/s12015-026-11215-4","DOIUrl":"https://doi.org/10.1007/s12015-026-11215-4","url":null,"abstract":"<p><p>Osteoporotic fracture is the most severe complication of osteoporosis, and impaired bone regeneration and delayed healing after fracture remain major clinical challenges. Extracellular vesicles (EVs) derived from bone marrow mesenchymal stem cells (BMSCs) may play a critical role in promoting bone regeneration after osteoporotic fracture. Enhancing their osteogenic potential and elucidating the underlying mechanisms may contribute to accelerated fracture healing. In this study, resveratrol-stimulated BMSC-derived EVs (Res-EVs) were isolated using differential centrifugation combined with ultracentrifugation. We found that compared with EVs secreted by BMSCs under normal culture conditions (Nor-EVs), Res-EVs significantly enhanced BMSCs osteogenic differentiation capacity. Analysis of miRNA sequencing data identified miR-19a-3p as significantly downregulated and PC-3p-83,608 as significantly upregulated. The reliability of the sequencing data was validated by qPCR results. By transfecting BMSCs with inhibitor and mimic of miR-19a-3p, we found that miR-19a-3p was a negative regulator of osteogenic differentiation. Bioinformatics prediction and dual-luciferase reporter assays confirmed that miR-19a-3p directly targets Raf1. Western blot results indicated that miR-19a-3p regulates osteogenic differentiation of BMSCs by targeting Raf1. In vivo experiments further demonstrated that Res-EVs treatment enhanced repair at osteoporotic fracture sites and increased the expression of RUNX2 and RAF1. Collectively, these findings indicate that Res-EVs promote bone regeneration at osteoporotic bone defect sites in rats, with the miR-19a-3p/Raf1 axis representing a potential underlying mechanism.</p>","PeriodicalId":21955,"journal":{"name":"Stem Cell Reviews and Reports","volume":" ","pages":""},"PeriodicalIF":4.9,"publicationDate":"2026-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148762206","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}