International journal of stem cells最新文献_第8页

Mimicking the Human Articular Joint with In Vitro Model of Neurons-Synoviocytes Co-Culture. 神经元-滑膜细胞体外共培养模拟人关节模型。

IF 2.3 4区医学

International journal of stem cells Pub Date : 2024-02-28 Epub Date: 2023-11-24 DOI: 10.15283/ijsc23043

Jakub Chwastek, Marta Kędziora, Małgorzata Borczyk, Michał Korostyński, Katarzyna Starowicz

{"title":"Mimicking the Human Articular Joint with In Vitro Model of Neurons-Synoviocytes Co-Culture.","authors":"Jakub Chwastek, Marta Kędziora, Małgorzata Borczyk, Michał Korostyński, Katarzyna Starowicz","doi":"10.15283/ijsc23043","DOIUrl":"10.15283/ijsc23043","url":null,"abstract":"The development of in vitro models is essential in modern science due to the need for experiments using human material and the reduction in the number of laboratory animals. The complexity of the interactions that occur in living organisms requires improvements in the monolayer cultures. In the work presented here, neuroepithelial stem (NES) cells were differentiated into peripheral-like neurons (PLN) and the phenotype of the cells was confirmed at the genetic and protein levels. Then RNA-seq method was used to investigate how stimulation with pro-inflammatory factors such as LPS and IFNγ affects the expression of genes involved in the immune response in human fibroblast-like synoviocytes (HFLS). HFLS were then cultured on semi-permeable membrane inserts, and after 24 hours of pro-inflammatory stimulation, the levels of cytokines secretion into the medium were checked. Inserts with stimulated HFLS were introduced into the PLN culture, and by measuring secreted ATP, an increase in cell activity was found in the system. The method used mimics the condition that occurs in the joint during inflammation, as observed in the development of diseases such as rheumatoid arthritis (RA) or osteoarthritis (OA). In addition, the system used can be easily modified to simulate the interaction of peripheral neurons with other cell types.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":" ","pages":"91-98"},"PeriodicalIF":2.3,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10899880/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138299072","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Host-Microbe Interactions Regulate Intestinal Stem Cells and Tissue Turnover in Drosophila. 宿主-微生物相互作用调控果蝇的肠干细胞和组织转换

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-12-21 DOI: 10.15283/ijsc23172

Ji-Hoon Lee

{"title":"Host-Microbe Interactions Regulate Intestinal Stem Cells and Tissue Turnover in Drosophila.","authors":"Ji-Hoon Lee","doi":"10.15283/ijsc23172","DOIUrl":"https://doi.org/10.15283/ijsc23172","url":null,"abstract":"With the activity of intestinal stem cells and continuous turnover, the gut epithelium is one of the most dynamic tissues in animals. Due to its simple yet conserved tissue structure and enteric cell composition as well as advanced genetic and histologic techniques, Drosophila serves as a valuable model system for investigating the regulation of intestinal stem cells. The Drosophila gut epithelium is in constant contact with indigenous microbiota and encounters externally introduced \"non-self\" substances, including foodborne pathogens. Therefore, in addition to its role in digestion and nutrient absorption, another essential function of the gut epithelium is to control the expansion of microbes while maintaining its structural integrity, necessitating a tissue turnover process involving intestinal stem cell activity. As a result, the microbiome and pathogens serve as important factors in regulating intestinal tissue turnover. In this manuscript, I discuss crucial discoveries revealing the interaction between gut microbes and the host's innate immune system, closely associated with the regulation of intestinal stem cell proliferation and differentiation, ultimately contributing to epithelial homeostasis.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"30 1","pages":""},"PeriodicalIF":2.3,"publicationDate":"2023-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138825034","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}

引用次数: 0

Differentiation and Characterization of Cystic Fibrosis Transmembrane Conductance Regulator Knockout Human Pluripotent Stem Cells into Salivary Gland Epithelial Progenitors. 囊性纤维化跨膜传导调节因子敲除人多能干细胞向唾液腺上皮祖细胞的分化和表征。

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-11-30 Epub Date: 2023-09-06 DOI: 10.15283/ijsc23036

Shuang Yan, Yifei Zhang, Siqi Zhang, Shicheng Wei

{"title":"Differentiation and Characterization of Cystic Fibrosis Transmembrane Conductance Regulator Knockout Human Pluripotent Stem Cells into Salivary Gland Epithelial Progenitors.","authors":"Shuang Yan, Yifei Zhang, Siqi Zhang, Shicheng Wei","doi":"10.15283/ijsc23036","DOIUrl":"10.15283/ijsc23036","url":null,"abstract":"The differentiation of pluripotent stem cells has been used to study disease mechanisms and development. We previously described a method for differentiating human pluripotent stem cells (hPSCs) into salivary gland epithelial progenitors (SGEPs). Here, cystic fibrosis transmembrane conductance regulator (CFTR) knockout hPSCs were differentiated into SGEPs derived from CFTR knockout hESCs (CF-SGEPs) using the same protocol to investigate whether the hPSC-derived SGEPs can model the characteristics of CF. CF-a disease that affects salivary gland (SG) function-is caused by mutations of the CFTR gene. Firstly, we successfully generated CFTR knockout hPSCs with reduced CFTR protein expression using the CRISPR-Cas9 system. After 16 days of differentiation, the protein expression of CFTR decreased in SGEPs derived from CFTR knockout hESCs (CF-SGEPs). RNA-Seq revealed that multiple genes modulating SG development and function were down-regulated, and positive regulators of inflammation were up-regulated in CF-SGEPs, correlating with the salivary phenotype of CF patients. These results demonstrated that CFTR suppression disrupted the differentiation of hPSC-derived SGEPs, which modeled the SG development of CF patients. In summary, this study not only proved that the hPSC-derived SGEPs could serve as manipulable and readily accessible cell models for the study of SG developmental diseases but also opened up new avenues for the study of the CF mechanism.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":" ","pages":"394-405"},"PeriodicalIF":2.3,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686799/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10218343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Corrigendum to "Cyclic Phytosphingosine-1-Phosphate Primed Mesenchymal Stem Cells Ameliorate LPS-Induced Acute Lung Injury in Mice". “环鞘氨醇-1-磷酸引发的间充质干细胞改善lps诱导的小鼠急性肺损伤”的勘误表。

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-11-30 DOI: 10.15283/23001C

Youngheon Park, Jimin Jang, Jooyeon Lee, Hyosin Baek, Jaehyun Park, Sang-Ryul Cha, Se Bi Lee, Sunghun Na, Jae-Woo Kwon, Young Jun Park, Myeong Jun Choi, Kye-Seong Kim, Seok-Ho Hong, Se-Ran Yang

引用次数: 0

RNF43 and ZNRF3 in Wnt Signaling - A Master Regulator at the Membrane. RNF43和ZNRF3在Wnt信号传导中的作用-膜上的主要调节因子。

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-11-30 Epub Date: 2023-08-30 DOI: 10.15283/ijsc23070

Fiona Farnhammer, Gabriele Colozza, Jihoon Kim

引用次数: 1

Adipose Tissue-Derived Mesenchymal Stromal Cells from Ex-Morbidly Obese Individuals Instruct Macrophages towards a M2-Like Profile In Vitro. 来自前病态肥胖个体的脂肪组织来源的间充质间质细胞在体外指导巨噬细胞向m2样结构发展

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-11-30 Epub Date: 2023-08-30 DOI: 10.15283/ijsc22172

Daiana V Lopes Alves, Cesar Claudio-da-Silva, Marcelo C A Souza, Rosa T Pinho, Wellington Seguins da Silva, Periela S Sousa-Vasconcelos, Radovan Borojevic, Carmen M Nogueira, Hélio Dos S Dutra, Christina M Takiya, Danielle C Bonfim, Maria Isabel D Rossi

{"title":"Adipose Tissue-Derived Mesenchymal Stromal Cells from Ex-Morbidly Obese Individuals Instruct Macrophages towards a M2-Like Profile In Vitro.","authors":"Daiana V Lopes Alves, Cesar Claudio-da-Silva, Marcelo C A Souza, Rosa T Pinho, Wellington Seguins da Silva, Periela S Sousa-Vasconcelos, Radovan Borojevic, Carmen M Nogueira, Hélio Dos S Dutra, Christina M Takiya, Danielle C Bonfim, Maria Isabel D Rossi","doi":"10.15283/ijsc22172","DOIUrl":"10.15283/ijsc22172","url":null,"abstract":"Obesity, which continues to increase worldwide, was shown to irreversibly impair the differentiation potential and angiogenic properties of adipose tissue mesenchymal stromal cells (ADSCs). Because these cells are intended for regenerative medicine, especially for the treatment of inflammatory conditions, and the effects of obesity on the immunomodulatory properties of ADSCs are not yet clear, here we investigated how ADSCs isolated from former obese subjects (Ex-Ob) would influence macrophage differentiation and polarization, since these cells are the main instructors of inflammatory responses. Analysis of the subcutaneous adipose tissue (SAT) of overweight (OW) and Ex-Ob subjects showed the maintenance of approximately twice as many macrophages in Ex-Ob SAT, contained within the CD68＋/FXIII-A- inflammatory pool. Despite it, in vitro, coculture experiments revealed that Ex-Ob ADSCs instructed monocyte differentiation into a M2-like profile, and under inflammatory conditions induced by LPS treatment, inhibited HLA-DR upregulation by resting M0 macrophages, originated a similar percentage of TNF-α＋ cells, and inhibited IL-10 secretion, similar to OW-ADSCs and BMSCs, which were used for comparison, as these are the main alternative cell types available for therapeutic purposes. Our results showed that Ex-Ob ADSCs mirrored OW-ADSCs in macrophage education, favoring the M2 immunophenotype and a mixed (M1/M2) secretory response. These results have translational potential, since they provide evidence that ADSCs from both Ex-Ob and OW subjects can be used in regenerative medicine in eligible therapies. Further in vivo studies will be fundamental to validate these observations.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":" ","pages":"425-437"},"PeriodicalIF":2.3,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686802/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10103748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In Vivo Stem Cell Imaging Principles and Applications. 体内干细胞成像原理和应用。

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-11-30 Epub Date: 2023-08-30 DOI: 10.15283/ijsc23045

Seongje Hong, Dong-Sung Lee, Geun-Woo Bae, Juhyeong Jeon, Hak Kyun Kim, Siyeon Rhee, Kyung Oh Jung

{"title":"In Vivo Stem Cell Imaging Principles and Applications.","authors":"Seongje Hong, Dong-Sung Lee, Geun-Woo Bae, Juhyeong Jeon, Hak Kyun Kim, Siyeon Rhee, Kyung Oh Jung","doi":"10.15283/ijsc23045","DOIUrl":"10.15283/ijsc23045","url":null,"abstract":"Stem cells are the foundational cells for every organ and tissue in our body. Cell-based therapeutics using stem cells in regenerative medicine have received attracting attention as a possible treatment for various diseases caused by congenital defects. Stem cells such as induced pluripotent stem cells (iPSCs) as well as embryonic stem cells (ESCs), mesenchymal stem cells (MSCs), and neuroprogenitors stem cells (NSCs) have recently been studied in various ways as a cell-based therapeutic agent. When various stem cells are transplanted into a living body, they can differentiate and perform complex functions. For stem cell transplantation, it is essential to determine the suitability of the stem cell-based treatment by evaluating the origin of stem, the route of administration, in vivo bio-distribution, transplanted cell survival, function, and mobility. Currently, these various stem cells are being imaged in vivo through various molecular imaging methods. Various imaging modalities such as optical imaging, magnetic resonance imaging (MRI), ultrasound (US), positron emission tomography (PET), and single-photon emission computed tomography (SPECT) have been introduced for the application of various stem cell imaging. In this review, we discuss the principles and recent advances of in vivo molecular imaging for application of stem cell research.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":" ","pages":"363-375"},"PeriodicalIF":2.3,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686800/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10103744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Transcriptional Signature of Valproic Acid-Induced Neural Tube Defects in Human Spinal Cord Organoids. 丙戊酸诱导的人脊髓类器官神经管缺损的转录特征。

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-11-30 Epub Date: 2023-08-30 DOI: 10.15283/ijsc23012

Ju-Hyun Lee, Mohammed R Shaker, Si-Hyung Park, Woong Sun

{"title":"Transcriptional Signature of Valproic Acid-Induced Neural Tube Defects in Human Spinal Cord Organoids.","authors":"Ju-Hyun Lee, Mohammed R Shaker, Si-Hyung Park, Woong Sun","doi":"10.15283/ijsc23012","DOIUrl":"10.15283/ijsc23012","url":null,"abstract":"In vertebrates, the entire central nervous system is derived from the neural tube, which is formed through a conserved early developmental morphogenetic process called neurulation. Although the perturbations in neurulation caused by genetic or environmental factors lead to neural tube defects (NTDs), the most common congenital malformation and the precise molecular pathological cascades mediating NTDs are not well understood. Recently, we have developed human spinal cord organoids (hSCOs) that recapitulate some aspects of human neurulation and observed that valproic acid (VPA) could cause neurulation defects in an organoid model. In this study, we identified and verified the significant changes in cell-cell junctional genes/proteins in VPA-treated organoids using transcriptomic and immunostaining analysis. Furthermore, VPA-treated mouse embryos exhibited impaired gene expression and NTD phenotypes, similar to those observed in the hSCO model. Collectively, our data demonstrate that hSCOs provide a valuable biological resource for dissecting the molecular pathways underlying the currently unknown human neurulation process using destructive biological analysis tools.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":" ","pages":"385-393"},"PeriodicalIF":2.3,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686804/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10112448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Assessment of Risks and Benefits of Using Antibiotics Resistance Genes in Mesenchymal Stem Cell-Based Ex-Vivo Therapy. 在间充质干细胞离体治疗中使用抗生素耐药基因的风险和益处评估。

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-11-30 Epub Date: 2023-06-30 DOI: 10.15283/ijsc23053

Narayan Bashyal, Young Jun Lee, Jin-Hwa Jung, Min Gyeong Kim, Kwang-Wook Lee, Woo Sup Hwang, Sung-Soo Kim, Da-Young Chang, Haeyoung Suh-Kim

{"title":"Assessment of Risks and Benefits of Using Antibiotics Resistance Genes in Mesenchymal Stem Cell-Based Ex-Vivo Therapy.","authors":"Narayan Bashyal, Young Jun Lee, Jin-Hwa Jung, Min Gyeong Kim, Kwang-Wook Lee, Woo Sup Hwang, Sung-Soo Kim, Da-Young Chang, Haeyoung Suh-Kim","doi":"10.15283/ijsc23053","DOIUrl":"10.15283/ijsc23053","url":null,"abstract":"Recently, ex-vivo gene therapy has emerged as a promising approach to enhance the therapeutic potential of mesenchymal stem cells (MSCs) by introducing functional genes in vitro. Here, we explored the need of using selection markers to increase the gene delivery efficiency and evaluated the potential risks associated with their use in the manufacturing process. We used MSCs/CD that carry the cytosine deaminase gene (CD) as a therapeutic gene and a puromycin resistance gene (PuroR) as a selection marker. We evaluated the correlation between the therapeutic efficacy and the purity of therapeutic MSCs/CD by examining their anti-cancer effect on co-cultured U87/GFP cells. To simulate in vivo horizontal transfer of the PuroR gene in vivo, we generated a puromycin-resistant E. coli (E. coli/PuroR) by introducing the PuroR gene and assessed its responsiveness to various antibiotics. We found that the anti-cancer effect of MSCs/CD was directly proportional to their purity, suggesting the crucial role of the PuroR gene in eliminating impure unmodified MSCs and enhancing the purity of MSCs/CD during the manufacturing process. Additionally, we found that clinically available antibiotics were effective in inhibiting the growth of hypothetical microorganism, E. coli/PuroR. In summary, our study highlights the potential benefits of using the PuroR gene as a selection marker to enhance the purity and efficacy of therapeutic cells in MSC-based gene therapy. Furthermore, our study suggests that the potential risk of horizontal transfer of antibiotics resistance genes in vivo can be effectively managed by clinically available antibiotics.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":" ","pages":"438-447"},"PeriodicalIF":2.3,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686797/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9690162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Preclinical Study on Biodistribution of Mesenchymal Stem Cells after Local Transplantation into the Brain. 间充质干细胞局部脑移植后生物分布的临床前研究。

IF 2.3 4区医学

International journal of stem cells Pub Date : 2023-11-30 Epub Date: 2023-08-30 DOI: 10.15283/ijsc23062

Narayan Bashyal, Min Gyeong Kim, Jin-Hwa Jung, Rakshya Acharya, Young Jun Lee, Woo Sup Hwang, Jung-Mi Choi, Da-Young Chang, Sung-Soo Kim, Haeyoung Suh-Kim

{"title":"Preclinical Study on Biodistribution of Mesenchymal Stem Cells after Local Transplantation into the Brain.","authors":"Narayan Bashyal, Min Gyeong Kim, Jin-Hwa Jung, Rakshya Acharya, Young Jun Lee, Woo Sup Hwang, Jung-Mi Choi, Da-Young Chang, Sung-Soo Kim, Haeyoung Suh-Kim","doi":"10.15283/ijsc23062","DOIUrl":"10.15283/ijsc23062","url":null,"abstract":"Therapeutic efficacy of mesenchymal stem cells (MSCs) is determined by biodistribution and engraftment in vivo. Compared to intravenous infusion, biodistribution of locally transplanted MSCs are partially understood. Here, we performed a pharmacokinetics (PK) study of MSCs after local transplantation. We grafted human MSCs into the brains of immune-compromised nude mice. Then we extracted genomic DNA from brains, lungs, and livers after transplantation over a month. Using quantitative polymerase chain reaction with human Alu-specific primers, we analyzed biodistribution of the transplanted cells. To evaluate the role of residual immune response in the brain, MSCs expressing a cytosine deaminase (MSCs/CD) were used to ablate resident immune cells at the injection site. The majority of the Alu signals mostly remained at the injection site and decreased over a week, finally becoming undetectable after one month. Negligible signals were transiently detected in the lung and liver during the first week. Suppression of Iba1-positive microglia in the vicinity of the injection site using MSCs/CD prolonged the presence of the Alu signals. After local transplantation in xenograft animal models, human MSCs remain predominantly near the injection site for limited time without disseminating to other organs. Transplantation of human MSCs can locally elicit an immune response in immune compromised animals, and suppressing resident immune cells can prolong the presence of transplanted cells. Our study provides valuable insights into the in vivo fate of locally transplanted stem cells and a local delivery is effective to achieve desired dosages for neurological diseases.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":" ","pages":"415-424"},"PeriodicalIF":2.3,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686801/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10109941","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0