检索结果-南通市图书馆

Predicting differentiation potential of human pluripotent stem cells:Possibilities and challenges

World Journal of Stem Cells 2019年第7期11卷 375-382页

作者： Li-Ping Liu yun-wen zheng Institute of Regenerative Medicine Affiliated Hospital of Jiangsu UniversityJiangsu UniversityZhenjiang 212001Jiangsu ProvinceChina University of Tsukuba Faculty of Medicine TsukubaIbaraki 305-8575Japan Yokohama City University School of Medicine Yokohama 236-0004Japan

The capability of human pluripotent stem cell(hPSC)lines to propagate indefinitely and differentiate into derivatives of three embryonic germ layers makes these cells be powerful tools for basic scientific research and promising agents for translational medicine.However,variations in differentiation tendency and efficiency as well as pluripotency maintenance necessitate the selection of hPSC lines for the intended applications to save time and cost.To screen the qualified cell lines and exclude problematic cell lines,their pluripotency must be confirmed initially by traditional methods such as teratoma formation or by highthroughput gene expression profiling assay.Additionally,their differentiation potential,particularly the lineage-specific differentiation propensities of hPSC lines,should be predicted in an early stage.As a complement to the teratoma assay,RNA sequencing data provide a quantitative estimate of the differentiation ability of hPSCs in vivo.Moreover,multiple scorecards have been developed based on selected gene sets for predicting the differentiation potential into three germ layers or the desired cell type many days before terminal differentiation.For clinical application of hPSCs,the malignant potential of the cells must also be evaluated.A combination of histologic examination of teratoma with quantitation of gene expression data derived from teratoma tissue provides safety-related predictive information by detecting immature teratomas,malignancy marker expression,and other parameters.Although various prediction methods are available,distinct limitations remain such as the discordance of results between different assays and requirement of a long time and high labor and cost,restricting their wide applications in routine studies.Therefore,simpler and more rapid detection assays with high specificity and sensitivity that can be used to monitor the status of hPSCs at any time and fewer targeted markers that are more specific for a given desired cell type are urgently needed.

关键词： Human pluripotent stem cells Induced pluripotent stem cells Embryonic stem cells Differentiation potential Prediction Pluripotency Malignant potential Embryoid bodies Lineage-specific differentiation Teratoma

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Hepatitis B virus infection modeling using multi-cellular organoids derived from human induced pluripotent stem cells

引用

World Journal of Gastroenterology 2021年第29期27卷 4784-4801页

作者： Di Cao Jian-yun Ge yun Wang Tatsuya Oda yun-wen zheng Institute of Regenerative Medicine and Affiliated Hospital of Jiangsu University Jiangsu UniversityZhenjiang 212001Jiangsu ProvinceChina Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery Faculty of MedicineUniversity of TsukubaTsukuba 305-8575IbarakiJapan Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine and School of Biotechnology and Heath SciencesWuyi UniversityJiangmen 529020Guangdong ProvinceChina School of Medicine Yokohama City UniversityYokohama 234-0006KanagawaJapan

Chronic infection with hepatitis B virus(HBV)remains a global health concern despite the availability of vaccines.To date,the development of effective treatments has been severely hampered by the lack of reliable,reproducible,and scalable in vitro modeling systems that precisely recapitulate the virus life cycle and represent virus-host interactions.With the progressive understanding of liver organogenesis mechanisms,the development of human induced pluripotent stem cell(iPSC)-derived hepatic sources and stromal cellular compositions provides novel strategies for personalized modeling and treatment of liver disease.Further,advancements in three-dimensional culture of self-organized liver-like organoids considerably promote in vitro modeling of intact human liver tissue,in terms of both hepatic function and other physiological characteristics.Combined with our experiences in the investigation of HBV infections using liver organoids,we have summarized the advances in modeling reported thus far and discussed the limitations and ongoing challenges in the application of liver organoids,particularly those with multi-cellular components derived from human iPSCs.This review provides general guidelines for establishing clinical-grade iPSC-derived multi-cellular organoids in modeling personalized hepatitis virus infection and other liver diseases,as well as drug testing and transplantation therapy.

关键词： Hepatitis B virus Induced pluripotent stem cells Liver organoid Multicellular organoid Modeling Transplantable

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Impelling force and current challenges by chemicals in somatic cell reprogramming and expansion beyond hepatocytes

引用

World Journal of Stem Cells 2019年第9期11卷 650-665页

作者： Jian-yun Ge yun-wen zheng Li-Ping Liu Hiroko Isoda Tatsuya Oda Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery Faculty of MedicineUniversity of TsukubaTsukubaIbaraki 305-8575Japan Institute of Regenerative Medicine and Affiliated Hospital Jiangsu UniversityZhenjiang 212001Jiangsu ProvinceChina Department of Regenerative Medicine School of MedicineYokohama City UniversityYokohama 236-0004Japan Faculty of Life and Environmental Sciences University of TsukubaTsukubaIbaraki 305-8572Japan

In the field of regenerative medicine,generating numerous transplantable functional cells in the laboratory setting on a large scale is a major challenge.However,the in vitro maintenance and expansion of terminally differentiated cells are challenging because of the lack of specific environmental and intercellular signal stimulations,markedly hindering their therapeutic application.Remarkably,the generation of stem/progenitor cells or functional cells with effective proliferative potential is markedly in demand for disease modeling,cell-based transplantation,and drug discovery.Despite the potent genetic manipulation of transcription factors,integration-free chemically defined approaches for the conversion of somatic cell fate have garnered considerable attention in recent years.This review aims to summarize the progress thus far and discuss the advantages,limitations,and challenges of the impact of full chemicals on the stepwise reprogramming of pluripotency,direct lineage conversion,and direct lineage expansion on somatic cells.Owing to the current chemical-mediated induction,reprogrammed pluripotent stem cells with reproducibility difficulties,and direct lineage converted cells with marked functional deficiency,it is imperative to generate the desired cell types directly by chemically inducing their potent proliferation ability through a lineagecommitted progenitor state,while upholding the maturation and engraftment capacity posttransplantation in vivo.Together with the comprehensive understanding of the mechanism of chemical drives,as well as the elucidation of specificity and commonalities,the precise manipulation of the expansion for diverse functional cell types could broaden the available cell sources and enhance the cellular function for clinical application in future.

关键词： Chemical induction Pluripotent reprogramming Direct lineage conversion Direct lineage expansion Hepatocyte expansion Cell fate specificity Transcriptional memory In vivo induction

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Inducing human induced pluripotent stem cell differentiation through embryoid bodies:A practical and stable approach

引用

World Journal of Stem Cells 2020年第1期12卷 25-34页

作者： Ning-Ning Guo Li-Ping Liu yun-wen zheng Yu-Mei Li Institute of Regenerative Medicine Affiliated Hospital of Jiangsu UniversityJiangsu UniversityZhenjiang 212001Jiangsu ProvinceChina Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery University of Tsukuba Faculty of MedicineTsukubaIbaraki 305-8575Japan Yokohama City University School of Medicine YokohamaKanagawa 234-0006Japan Division of Regenerative Medicine Center for Stem Cell Biology and Regenerative MedicineThe Institute of Medical Sciencethe University of TokyoTokyo 108-8639Japan

Human induced pluripotent stem cells(hiPSCs)are invaluable resources for producing high-quality differentiated cells in unlimited quantities for both basic research and clinical use.They are particularly useful for studying human disease mechanisms in vitro by making it possible to circumvent the ethical issues of human embryonic stem cell research.However,significant limitations exist when using conventional flat culturing methods especially concerning cell expansion,differentiation efficiency,stability maintenance and multicellular 3D structure establishment,differentiation prediction.Embryoid bodies(EBs),the multicellular aggregates spontaneously generated from iPSCs in the suspension system,might help to address these issues.Due to the unique microenvironment and cell communication in EB structure that a 2D culture system cannot achieve,EBs have been widely applied in hiPSC-derived differentiation and show significant advantages especially in scaling up culturing,differentiation efficiency enhancement,ex vivo simulation,and organoid establishment.EBs can potentially also be used in early prediction of iPSC differentiation capability.To improve the stability and feasibility of EB-mediated differentiation and generate high quality EBs,critical factors including iPSC pluripotency maintenance,generation of uniform morphology using micro-pattern 3D culture systems,proper cellular density inoculation,and EB size control are discussed on the basis of both published data and our own laboratory experiences.Collectively,the production of a large quantity of homogeneous EBs with high quality is important for the stability and feasibility of many PSCs related studies.

关键词： Induced pluripotent stem cells Suspension culture Embryoid body Early prediction Committed differentiation Heterogeneity Three-dimensional culture Scaling-up Quality control

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Practical choice for robust and efficient differentiation of human pluripotent stem cells

引用

World Journal of Stem Cells 2020年第8期12卷 752-760页

作者： Mei Fang Li-Ping Liu Hang Zhou Yu-Mei Li yun-wen zheng Institute of Regenerative Medicine Affiliated Hospital of Jiangsu UniversityJiangsu UniversityZhenjiang 212001Jiangsu ProvinceChina School of Biotechnology and Heath Sciences Wuyi UniversityJiangmen 529020Guangdong ProvinceChina Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery University of Tsukuba Faculty of MedicineTsukubaIbaraki 305-8575Japan City University School of Medicine YokohamaKanagawa 234-0006Japan Division of Regenerative Medicine Center for Stem Cell Biology and Regenerative MedicineThe Institute of Medical Sciencethe University of TokyoTokyo 108-8639Japan

Human pluripotent stem cells(hPSCs)have the distinct advantage of being able to differentiate into cells of all three germ layers.Target cells or tissues derived from hPSCs have many uses such as drug screening,disease modeling,and transplantation therapy.There are currently a wide variety of differentiation methods available.However,most of the existing differentiation methods are unreliable,with uneven differentiation efficiency and poor reproducibility.At the same time,it is difficult to choose the optimal method when faced with so many differentiation schemes,and it is time-consuming and costly to explore a new differentiation approach.Thus,it is critical to design a robust and efficient method of differentiation.In this review article,we summarize a comprehensive approach in which hPSCs are differentiated into target cells or organoids including brain,liver,blood,melanocytes,and mesenchymal cells.This was accomplished by employing an embryoid body-based three-dimensional(3D)suspension culture system with multiple cells co-cultured.The method has high stable differentiation efficiency compared to the conventional 2D culture and can meet the requirements of clinical application.Additionally,ex vivo co-culture models might be able to constitute organoids that are highly similar or mimic human organs for potential organ transplantation in the future.

关键词： Human pluripotent stem cells Three dimensional Embryoid body Differentiation Efficient Three germ layers

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Novel alternative transplantation therapy for orthotopic liver transplantation in liver failure:A systematic review

引用

World Journal of Transplantation 2020年第3期10卷 64-78页

作者： Tomoaki Furuta Kinji Furuya yun-wen zheng Tatsuya Oda Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery Faculty of MedicineUniversity of TsukubaTsukuba-shi 305-8575IbarakiJapan Institute of Regenerative Medicine and Affiliated Hospital of Jiangsu University Zhenjiang 212001Jiangsu ProvinceChina Department of Regenerative Medicine School of MedicineYokohama City UniversityYokohama 236-0004Japan Center for Stem Cell Biology and Regenerative Medicine The Institute of Medical ScienceThe University of TokyoTokyo 108-8639Japan

BACKGROUND Orthotopic liver transplantation(OLT)is the only treatment for end-stage liver failure;however,graft shortage impedes its applicability.Therefore,studies investigating alternative therapies are plenty.Nevertheless,no study has comprehensively analyzed these therapies from different perspectives.AIM To summarize the current status of alternative transplantation therapies for OLT and to support future research.METHODS A systematic literature search was performed using PubMed,Cochrane Library and EMBASE for articles published between January 2010 and 2018,using the following MeSH terms:[(liver transplantation)AND cell]OR[(liver transplantation)AND differentiation]OR[(liver transplantation)AND organoid]OR[(liver transplantation)AND xenotransplantation].Various types of studies describing therapies to replace OLT were retrieved for full-text evaluation.Among them,we selected articles including in vivo transplantation.RESULTS A total of 89 studies were selected.There are three principle forms of treatment for liver failure:Xeno-organ transplantation,scaffold-based transplantation,and cell transplantation.Xeno-organ transplantation was covered in 14 articles,scaffold-based transplantation was discussed in 22 articles,and cell transplantation was discussed in 53 articles.Various types of alternative therapies were discussed:Organ liver,25 articles;adult hepatocytes,31 articles;fetal hepatocytes,three articles;mesenchymal stem cells(MSCs),25 articles;embryonic stem cells,one article;and induced pluripotent stem cells,three articles and other sources.Clinical applications were discussed in 12 studies:Cell transplantation using hepatocytes in four studies,five studies using umbilical cord-derived MSCs,three studies using bone marrow-derived MSCs,and two studies using hematopoietic stem cells.CONCLUSION The clinical applications are present only for cell transplantation.Scaffold-based transplantation is a comprehensive treatment combining organ and cell transplantations,which warrants future research to find relev

关键词： Cell transplantation Liver transplantation Organ transplantation Xenotransplantation Tissue engineering Scaffold

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Current trends in three-dimensional visualization and real-time navigation as well as robot-assisted technologies in hepatobiliary surgery

引用

World Journal of Gastrointestinal Surgery 2021年第9期13卷 904-922页

作者： yun Wang Di Cao Si-Lin Chen Yu-Mei Li yun-wen zheng Nobuhiro Ohkohchi Institute of Regenerative Medicine and Affiliated Hospital of Jiangsu UniversityJiangsu UniversityZhenjiang 212001Jiangsu ProvinceChina Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery Faculty of MedicineUniversity of TsukubaTsukuba 305-8575IbarakiJapan Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine and School of Biotechnology and Heath SciencesWuyi UniversityJiangmen 529020Guangdong ProvinceChina School of Medicine Yokohama City UniversityYokohama 234-0006KanagawaJapan

With the continuous development of digital medicine,minimally invasive precision and safety have become the primary development trends in hepatobiliary surgery.Due to the specificity and complexity of hepatobiliary surgery,traditional preoperative imaging techniques such as computed tomography and magnetic resonance imaging cannot meet the need for identification of fine anatomical regions.Imaging-based three-dimensional(3D)reconstruction,virtual simulation of surgery and 3D printing optimize the surgical plan through preoperative assessment,improving the controllability and safety of intraoperative operations,and in difficult-to-reach areas of the posterior and superior liver,assistive robots reproduce the surgeon’s natural movements with stable cameras,reducing natural vibrations.Electromagnetic navigation in abdominal surgery solves the problem of conventional surgery still relying on direct visual observation or preoperative image assessment.We summarize and compare these recent trends in digital medical solutions for the future development and refinement of digital medicine in hepatobiliary surgery.

关键词： Hepatobiliary surgery Three-dimensional visualization Three-dimensional printing Electromagnetic tracking Real-time navigation Robot-assisted surgery

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Enhanced hepatic differentiation in the subpopulation of human amniotic stem cells under 3D multicellular microenvironment

引用

World Journal of Stem Cells 2019年第9期11卷 705-721页

作者： Kinji Furuya yun-wen zheng Daisuke Sako Kenichi Iwasaki Dong-Xu zheng Jian-yun Ge Li-Ping Liu Tomoaki Furuta Kazunori Akimoto Hiroya Yagi Hiromi Hamada Hiroko Isoda Tatsuya Oda Nobuhiro Ohkohchi Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery Faculty of MedicineUniversity of TsukubaTsukubaIbaraki 305-8575Japan Institute of Regenerative Medicine and Affiliated Hospital Jiangsu UniversityZhenjiang 212001Jiangsu ProvinceChina Department of Regenerative Medicine School of MedicineYokohama City UniversityYokohama 236-0004Japan Department of Medicinal and Life Sciences Faculty of Pharmaceutical SciencesTokyo University of ScienceNoda 278-8510Japan Department of Obstetrics and Gynecology Faculty of MedicineUniversity of TsukubaTsukuba 305-8575Japan Faculty of Life and Environmental Sciences University of TsukubaTsukuba 305-8572Japan

BACKGROUND To solve the problem of liver transplantation donor insufficiency,an alternative cell transplantation therapy was investigated.We focused on amniotic epithelial cells(AECs)as a cell source because,unlike induced pluripotent stem cells,they are cost-effective and non-tumorigenic.The utilization of AECs in regenerative medicine,however,is in its infancy.A general profile for AECs has not been comprehensively analyzed.Moreover,no hepatic differentiation protocol for AECs has yet been established.To this end,we independently compiled human AEC libraries,purified amniotic stem cells(ASCs),and co-cultured them with mesenchymal stem cells(MSCs)and human umbilical vein endothelial cell(HUVECs)in a 3D system which induces functional hepatic organoids.AIM To characterize AECs and generate functional hepatic organoids from ASCs and other somatic stem cells METHODS AECs,MSCs,and HUVECs were isolated from the placentae and umbilical cords of cesarean section patients.Amnion and primary AEC stemness characteristics and heterogeneity were analyzed by immunocytochemistry,Alkaline phosphatase(AP)staining,and flow cytometry.An adherent AEC subpopulation was selected and evaluated for ASC purification quality by a colony formation assay.AEC transcriptomes were compared with those for other hepatocytes cell sources by bioinformatics.The 2D and 3D culture were compared by relative gene expression using several differentiation protocols.ASCs,MSCs,and HUVECs were combined in a 3D co-culture system to generate hepatic organoids whose structure was compared with a 3D AEC sphere and whose function was elucidated by immunofluorescence imaging,periodic acid Schiff,and an indocyanine green(ICG)test.RESULTS AECs have certain stemness markers such as EPCAM,SSEA4,and E-cadherin.One AEC subpopulation was also either positive for AP staining or expressed the TRA-1-60 and TRA-1-81 stemness markers.Moreover,it could form colonies and its frequency was enhanced ten-fold in the adherent subpopulation after selective primary passage.Bioinfo

关键词： 3D micropattern Amniotic epithelial cells Amniotic stem cells Hepatic differentiation Heterogeneity Human placental tissue Human umbilical vein endothelial cells Mesenchymal stem cells Multicellular microenvironment Organoid

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Quantitative evaluation of long-term liver repopulation and the reconstitution of bile ductules after hepatocellular transplantation

引用

World Journal of Gastroenterology 2005年第39期11卷 6176-6181页

作者： yun-wen zheng Nobuhiro Ohkohchi Hideki Taniguchi Department of Regenerative Medicine Graduate School of Medical Science Yokohama City University Yokohama 236-0004 Japan Department of Surgery Institute of Clinical Medicine University of Tsukuba Tsukuba 305-8575 Japan Research Unit for Organ Regeneration Center for Developmental Biology RIKEN Kobe 650-0047 Japan

AIM: The treatment of liver disease is severely limited by a shortage of donor livers. In trying to address this growing problem, hepatocellular transplantation (HTx) has received much attention as an alternative to whole organ transplant.However, the expansion of transplanted cells is at low level,and the reconstitution of functional liver tissue is limited by this cellular property. We set up an animal model to better understand cell dose effect and the kinetics of liver repopulation following HTx.METHODS: Dipeptidyl peptidase Ⅳ (DPPIV)-deficient rats treated with retrorsine and subjected to partial hepatectomy were infused with DPPIV-positive hepatocytes. Rats were injected with varying numbers of donor hepatocytes down to 100 cells low, and liver repopulation was examined at different time points up to 20 mo long. Repopulation was assessed by computer-aided quantitative detection.RESULTS: Transplanted hepatocytes underwent multiple rounds of proliferation and stably repopulated the injured livers after 20 mo and at all cell doses. Transplanted cells divided 14 times within the 3-mo time period following infusion, and the liver repopulation reached a plateau between 3 and 20 mo. Approximately 90% replacement occurred. Donor-derived cells also reconstituted the bile ductules of the recipients.CONCLUSION: The ability of transplanted hepatocytes to fully reconstitute injured livers strongly supports further investigation into the clinical potential of HTx. Additionally,the observation that transplanted hepatocytes also form components of the biliary system suggests that these cells may have bi-potential property of the stem cells.

关键词：定量检测胆管肝细胞移植检测方法病理机制

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Cellular reprogramming and hepatocellular carcinoma development

引用

World Journal of Gastroenterology 2013年第47期19卷 8850-8860页

作者： yun-wen zheng yun-Zhong Nie Hideki Taniguchi Department of Regenerative Medicine Graduate School of MedicineYokohama City UniversityKanazawa-kuYokohamaKanagawa 236-0004Japan Jiangsu University Hospital Advanced Medical Research Center Yokohama City UniversityYokohamaKanagawa 236-0004Japan

Hepatocellular carcinoma(HCC)is one of the most common cancers,and is also the leading cause of death worldwide.Studies have shown that cellular reprogramming contributes to chemotherapy and/or radiotherapy resistance and the recurrence of cancers.In this article,we summarize and discuss the latest findings in the area of cellular reprogramming in HCC.The aberrant expression of transcription factors OCT4,KLF4,SOX2,c-MYC,NANOG,and LIN28 have been also observed,and the expression of these transcription factors is associated with unfavorable clinical outcomes in HCC.Studies indicate that cellular reprogramming may play a critical role in the occurrence and recurrence of HCC.Recent reports have shown that DNA methylation,miRNAs,tumor microenvironment,and signaling pathways can induce the expression of stemness transcription factors,which leads to cellular reprogramming in HCC.Furthermore,studies indicate that therapies based on cellular reprogramming could revolutionize HCC treatment.Finally,a novel therapeutic concept is discussed:reprogramming control therapy.A potential reprogramming control therapy method could be developed based on the reprogramming demonstrated in HCC studies and applied at two opposing levels:differentiation and reprogramming.Our increasing understanding and control of cellular programming should facilitate the exploitation of this novel therapeutic concept and its application in clinical HCC treatment,which may represent a promising strategy in the future that is not restricted to liver cancer.

关键词： Reprogramming Hepatocellular carcinomas Cancer stem cells Transcription factor Therapeutics

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：