检索结果-内蒙古大学图书馆

conditional reprogramming culture conditions facilitate growth of lower-grade glioma models

NEURO-ONCOLOGY 2021年第5期23卷 770-782页

作者： Yuan, Ming White, David Resar, Linda Bar, Eli Groves, Mari Cohen, Alan Jackson, Eric Bynum, Jennifer Rubens, Jeffrey Mumm, Jeff Chen, Liam Jiang, Liqun Raabe, Eric Rodriguez, Fausto J. Eberhart, Charles G. Johns Hopkins Univ Dept Pathol Sch Med 720 Rutland AveRoss Bldg 558 Baltimore MD 21205 USA Johns Hopkins Univ Dept Ophthalmol Sch Med Baltimore MD 21205 USA Johns Hopkins Univ Div Pediat Oncol Sch Med Baltimore MD 21205 USA Univ Maryland Dept Pathol Baltimore MD 21201 USA Johns Hopkins Univ Sch Med Dept Neurosurg Baltimore MD 21205 USA

Background. The conditional reprogramming cell culture method was developed to facilitate growth of senescence-prone normal and neoplastic epithelial cells, and involves co-culture with irradiated fibroblasts and the addition of a small molecule Rho kinase (ROCK) inhibitor. The aim of this study was to determine whether this approach would facilitate the culture of compact low-grade gliomas. Methods. We attempted to culture 4 pilocytic astrocytomas, 2 gangliogliomas, 2 myxopapillary ependymomas, 2 anaplastic gliomas, 2 difficult-to-classify low-grade neuroepithelial tumors, a desmoplastic infantile ganglioglioma, and an anaplastic pleomorphic xanthoastrocytoma using a modified conditional reprogramming cell culture approach. Results. conditional reprogramming resulted in robust increases in growth for a majority of these tumors, with fibroblast conditioned media and ROCK inhibition both required. Switching cultures to standard serum containing media, or serum-free neurosphere conditions, with or without ROCK inhibition, resulted in decreased proliferation and induction of senescence markers. Rho kinase inhibition and conditioned media both promoted Akt and Erk1/2 activation. Several cultures, including one derived from a NF1-associated pilocytic astrocytoma (JHH-NF1-PA1) and one from a BRAF p.V600E mutant anaplastic pleomorphic xanthoastrocytoma (JHH-PXA1), exhibited growth sufficient for preclinical testing in vitro. In addition, JHH-NF1-PA1 cells survived and migrated in larval zebrafish orthotopic xenografts, while JHH-PXA1 formed orthotopic xenografts in mice histopathologically similar to the tumor from which it was derived. Conclusions. These studies highlight the potential for the conditional reprogramming cell culture method to promote the growth of glial and glioneuronal tumors in vitro, in some cases enabling the establishment of long-term culture and in vivo models.

关键词： BRAFV600E conditional reprogramming low-grade glioma NF1 enescence

来源：评论

学校读者我要写书评

暂无评论

conditional reprogramming: Modeling urological cancer and translation to clinics

引用

CLINICAL AND TRANSLATIONAL MEDICINE 2020年第2期10卷 e95页

作者： Liu, Wei Ju, Lingao Cheng, Songtao Wang, Gang Qian, Kaiyu Liu, Xuefeng Xiao, Yu Wang, Xinghuan Wuhan Univ Zhongnan Hosp Dept Urol Wuhan 430071 Peoples R China Wuhan Univ Zhongnan Hosp Dept Biol Repositories Wuhan Peoples R China Human Genet Resources Preservat Ctr Hubei Prov Wuhan Peoples R China Georgetown Univ Med Ctr Dept Pathol Lombardi Comprehens Canc Ctr Washington DC 20007 USA Wuhan Univ Med Res Inst Wuhan Peoples R China

Patient-derived models, including cell models (organoids and conditionally reprogrammed cells [CRCs]) and patient-derived xenografts, are urgently needed for both basic and translational cancer research. conditional reprogramming (CR) technique refers to a co-culture system of primary human normal or tumor cells with irradiated murine fibroblasts in the presence of a Rho-associated kinase inhibitor to allow the primary cells to acquire stem cell properties and the ability to proliferate indefinitely in vitro without any exogenous gene or viral transfection. Considering its robust features, the CR technique may facilitate cancer research in many aspects. Under in vitro culturing, malignant CRCs can share certain genetic aberrations and tumor phenotypes with their parental specimens. Thus, tumor CRCs can promisingly be utilized for the study of cancer biology, the discovery of novel therapies, and the promotion of precision medicine. For normal CRCs, the characteristics of normal karyotype maintenance and lineage commitment suggest their potential in toxicity testing and regenerative medicine. In this review, we discuss the applications, limitations, and future potential of CRCs in modeling urological cancer and translation to clinics.

关键词： conditional reprogramming patient-derived model precision medicine urological cancer

来源：评论

学校读者我要写书评

暂无评论

Establishment of a diverse head and neck squamous cancer cell bank using conditional reprogramming culture methods

引用

JOURNAL OF MEDICAL VIROLOGY 2023年第2期95卷 e28388-e28388页

作者： Li, Daniel Thomas, Carlos Shrivastava, Nitisha Gersten, Adam Gadsden, Nicholas Schlecht, Nicolas Kawachi, Nicole Schiff, Bradley A. A. Smith, Richard V. V. Rosenblatt, Gregory Augustine, Stelby Gavathiotis, Evripidis Burk, Robert Prystowsky, Michael B. B. Guha, Chandan Mehta, Vikas Ow, Thomas J. J. Albert Einstein Coll Med Montefiore Med Ctr Dept Pathol Bronx NY USA Columbia Univ Dept Anesthesiol New York NY USA Roswell Pk Comprehens Canc Ctr Dept Canc Prevent & Control Buffalo NY USA Albert Einstein Coll Med Montefiore Med Ctr Dept Otorhinolaryngol Head & Neck Surg Bronx NY USA Albert Einstein Coll Med Montefiore Med Ctr Dept Surg Bronx NY USA Albert Einstein Coll Med Dept Biochem Bronx NY USA Albert Einstein Coll Med Montefiore Med Ctr Dept Med Cardiol Bronx NY USA Albert Einstein Coll Med Montefiore Med Ctr Dept Pediat Bronx NY USA Albert Einstein Coll Med Montefiore Med Ctr Dept Microbiol & Immunol Bronx NY USA Albert Einstein Coll Med Montefiore Med Ctr Dept Epidemiol & Populat Hlth Epidemiol Bronx NY 10467 USA Albert Einstein Coll Med Montefiore Med Ctr Dept Radiat Oncol Bronx NY USA Albert Einstein Coll Med Montefiore Med Ctr Dept Urol Bronx NY USA

Most laboratory models of head and neck squamous cell cancer (HNSCC) rely on established immortalized cell lines, which carry inherent bias due to selection and clonality. We established a robust panel of HNSCC tumor cultures using a "conditional reprogramming" (CR) method, which utilizes a rho kinase inhibitor (Y-27632) and co-culture with irradiated fibroblast (J2 strain) feeder cells to support indefinite tumor cell survival. Sixteen CR cultures were successfully generated from 19 consecutively enrolled ethnically and racially diverse patients with HNSCC at a tertiary care center in the Bronx, NY. Of the 16 CR cultures, 9/16 were derived from the oral cavity, 4/16 were derived from the oropharynx, and 3/16 were from laryngeal carcinomas. Short tandem repeat (STR) profiling was used to validate culture against patient tumor tissue DNA. All CR cultures expressed & UDelta;Np63 and cytokeratin 5/6, which are markers of squamous identity. Human papillomavirus (HPV) testing was assessed utilizing clinical p16 staining on primary tumors, reverse transcription polymerase chain reaction (RT-PCR) of HPV16/18-specific viral oncogenes E6 and E7 in RNA extracted from tumor samples, and HPV DNA sequencing. Three of four oropharyngeal tumors were p16 and HPV-positive and maintained HPV in culture. CR cultures were able to establish three-dimensional spheroid and murine flank and orthotopic tongue models. CR methods can be readily applied to all HNSCC tumors regardless of patient characteristics, disease site, and molecular background, providing a translational research model that properly includes patient and tumor diversity.

关键词： animals cell culture techniques conditional reprogramming DNA fingerprinting head and neck neoplasms humans precision medicine primary cell culture spheroids squamous cell carcinoma of head and neck tissue banks whole exome sequencing

来源：评论

学校读者我要写书评

暂无评论

Rapid screening for individualized chemotherapy optimization of colorectal cancer: A novel conditional reprogramming technology-based functional diagnostic assay

引用

TRANSLATIONAL ONCOLOGY 2021年第1期14卷 100935-100935页

作者： Li, Yingjie Guo, Dagang Zhang, Yihong Wang, Lin Sun, Tingting Li, Zhongwu Zhang, Xiaoyan Wang, Shuai Chen, Yiyou Wu, Aiwen Peking Univ Canc Hosp & Inst Dept Gastrointestinal Surg Key Lab Carcinogenesis & Translat Res Minist Educ 52 Fucheng Rd Beijing 100142 Peoples R China Beijing Percans Oncol Res Co Ltd Bldg 115th FloorPKUCare Ind PkLife Sci Pk Beijing 102206 Peoples R China Peking Univ Dept Pathol Key Lab Carcinogenesis & Translat Res Minist EducSch OncolBeijing Canc Hosp & Inst Beijing 100142 Peoples R China Peking Univ Dept Radiol Key Lab Carcinogenesis & Translat Res Minist EducSch OncolBeijing KeyCanc Hosp & Inst Beijing 100142 Peoples R China

Background: In vitro patient tumor models such as patient-derived organoids (PDO) and conditionally reprogrammed (CR) cell culture are important for translational research and pre-clinical drug testing. In this study we present a personalized drug sensitivity test for late stage, potentially operable colorectal cancer (CRC) using patient-derived primary tumor cells isolated with i-CR technology, an optimized CR method. We explored the clinical feasibility of using i-CR platform to guide CRC chemotherapy, and established the correlation between in vitro drug sensitivity and patient clinical response. Methods: Primary CRC tumor cells were isolated and cultured with the i-CR technology. NGS was performed and the WES and CNV results of i-CR cells were compared with that of the original patient tumor samples. In vitro drug screenings were done with guideline chemotherapy drugs for CRC. In vivo drug response was examined with paired PDX mouse models. A double-blind co-clinical cohort study was carried out and the clinical outcomes of the enrolled patients were compared with the i-CR results. Results: i-CR platform could be used to rapidly propagate primary colorectal tumor cells that represent individual patient tumors effectively by keeping the clonal heterogeneity and the genetic characteristics. Chemotherapy drug screenings with i-CR cells were comparable with that of PDX models. More importantly, i-CR results showed high accordance with the clinical outcomes of the enrolled CRC patients. Conclusion: i-CR platform was capable to test and optimize therapeutic regimens pre-clinically, study cancer cell biology, and model tumor re-emergence to identify new targeted therapeutics from an effective personalized medicine standpoint.

关键词： conditional reprogramming Colorectal cancer Chemotherapy Pre-clinical In vitro drug test

来源：评论

学校读者我要写书评

暂无评论

conditional cell reprogramming for modeling host-virus interactions and human viral diseases

引用

JOURNAL OF MEDICAL VIROLOGY 2020年第11期92卷 2440-2452页

作者： Liu, Xuefeng Mondal, Abdul M. Georgetown Univ Med Ctr Dept Pathol Ctr Cell Reprogramming Washington DC 20007 USA Georgetown Univ Med Ctr Dept Oncol Lombardi Comprehens Canc Ctr Washington DC 20007 USA

Conventional cancer and transformed cell lines are widely used in cancer biology and other fields within biology. These cells usually have abnormalities from the original tumor itself, but may also develop abnormalities due to genetic manipulation, or genetic and epigenetic changes during long-term passages. Primary cultures may maintain lineage functions as the original tissue types, yet they have a very limited life span or population doubling time because of the nature of cellular senescence. Primary cultures usually have very low yields, and the high variability from any original tissue specimens, largely limiting their applications in research. Animal models are often used for studies of virus infections, disease modeling, development of antiviral drugs, and vaccines. Human viruses often need a series of passages in vivo to adapt to the host environment because of variable receptors on the cell surface and may have intracellular restrictions from the cell types or host species. Here, we describe a long-term cell culture system, conditionally reprogrammed cells (CRCs), and its applications in modeling human viral diseases and drug discovery. Using feeder layer coculture in presence of Y-27632 (conditional reprogramming, CR), CRCs can be obtained and rapidly propagated from surgical specimens, core or needle biopsies, and other minimally invasive or noninvasive specimens, for example, nasal cavity brushing. CRCs preserve their lineage functions and provide biologically relevant and physiological conditions, which are suitable for studies of viral entry and replication, innate immune responses of host cells, and discovery of antiviral drugs. In this review, we summarize the applications of CR technology in modeling host-virus interactions and human viral diseases including severe acute respiratory syndrome coronavirus-2 and coronavirus disease-2019, and antiviral discovery.

关键词： air-liquid interface cell senescence conditional reprogramming emerging viruses functional models normal cells organoids physiological conditions

来源：评论

学校读者我要写书评

暂无评论

Long-term culture and characterization of patient-derived primary hepatocytes using conditional reprogramming

引用

Experimental biology and medicine (Maywood, N.J.) 2019年第11期244卷 857-864页

作者： Shan Su Cristina Di Poto Rabindra Roy Xuefeng Liu Wanxing Cui Alexander Kroemer Habtom W Ressom 1 Lombardi Comprehensive Cancer Center Georgetown University Medical Center Washington DC 20057 USA. 2 Department of Pathology Georgetown University Medical Center Washington DC 20057 USA. 3 MedStar Georgetown University Hospital Washington DC 20057 USA. 4 MedStar Georgetown Transplant Institute Washington DC 20057 USA.

Cultivation of primary human hepatocytes (PHHs) often faces obstacles including failure of long-term in vitro culture, weak proliferation ability, rapid loss of liver-specific function and morphology, and tendency of fibrosis. Previous research focused on immortalization methods, such as telomerase and viral, to culture immortalized primary human hepatocytes, which may lose some of the normal properties. However, non-immortalized PHHs often fail to maintain long-term viability and functionality. These highlight the urgent need for developing new culture strategy for PHHs. In the present study, we isolated PHHs from fresh human liver tissues representing different liver diseases and age groups. We used conditional reprogramming, without permanent immortalization, for long-term in vitro primary human hepatocytes cultivation and characterization. For functional characterization, we assessed CYP3A4, 1A1 and 2C9 activities and measured the mRNA expression of albumin, s100a4, krt8, krt18, cyp1a1, cyp3a4, cyp2b6, cyp2c8, cyp2c9, and cyp2d6. Additionally, we compared the DNA fingerprint of the cells against their original liver tissues using short tandem repeat (STR) analysis. We found that PHHs-derived from young patients can survive for more than three months, while the lifespan of primary human hepatocytes derived from adult patients ranges from two to three months, which is longer than most commercial primary hepatocytes. Importantly, the cells at early passages retain strong CYP3A4, 1A1 and 2C9 activities and the DNA fingerprints are identical with their original tissues. Through conditional programming, we achieved, for the first time, a high level of success rate in the long-term in vitro cultivation of primary human hepatocytes-derived patients representing diverse liver disease. Moreover, the conditional programming cell culture technology reported in this paper requires neither co-culture with additive cells, nor complex and expensive components, such as collagen

关键词： Primary human hepatocytes conditional reprogramming long-term culture

来源：评论

学校读者我要写书评

暂无评论

Generation of stable PDX derived cell lines using conditional reprogramming

引用

MOLECULAR CANCER 2017年第20171228期16卷 177-177页

作者： Borodovsky, Alexandra McQuiston, Travis J. Stetson, Daniel Ahmed, Ambar Whitston, David Zhang, Jingwen Grondine, Michael Lawson, Deborah Challberg, Sharon S. Zinda, Michael Pollok, Brian A. Dougherty, Brian A. D'Cruz, Celina M. AstraZeneca IMED Biotech Unit Biosci Oncol Boston MA 02130 USA Propagenix Inc 9605 Med Ctr Dr 325 Rockville MD 20850 USA

Efforts to develop effective cancer therapeutics have been hindered by a lack of clinically predictive preclinical models which recapitulate this complex disease. Patient derived xenograft (PDX) models have emerged as valuable tools for translational research but have several practical limitations including lack of sustained growth in vitro. In this study, we utilized conditional reprogramming (CR) cell technology- a novel cell culture system facilitating the generation of stable cultures from patient biopsies-to establish PDX-derived cell lines which maintain the characteristics of the parental PDX tumor. Human lung and ovarian PDX tumors were successfully propagated using CR technology to create stable explant cell lines (CR-PDX). These CR-PDX cell lines maintained parental driver mutations and allele frequency without clonal drift. Purified CR-PDX cell lines were amenable to high throughput chemosensitivity screening and in vitro genetic knockdown studies. Additionally, re-implanted CR-PDX cells proliferated to form tumors that retained the growth kinetics, histology, and drug responses of the parental PDX tumor. CR technology can be used to generate and expand stable cell lines from PDX tumors without compromising fundamental biological properties of the model. It offers the ability to expand PDX cells in vitro for subsequent 2D screening assays as well as for use in vivo to reduce variability, animal usage and study costs. The methods and data detailed here provide a platform to generate physiologically relevant and predictive preclinical models to enhance drug discovery efforts.

关键词： Patient derived xenograft conditional reprogramming Cell line models Drug discovery Oncology

来源：评论

学校读者我要写书评

暂无评论

Establishment of a conditionally reprogrammed primary eccrine sweat gland culture for evaluation of tissue-specific CFTR function

引用

JOURNAL OF CYSTIC FIBROSIS 2024年第6期23卷 1173-1179页

作者： Eastman, Alice C. Rosson, Gedge Kim, Noori Kang, Sewon Raraigh, Karen Goff, Loyal A. Merlo, Christian Lechtzin, Noah Cutting, Garry R. Sharma, Neeraj Johns Hopkins Sch Med Dept Human Genet Miller Res Bldg 733N Broadway Baltimore MD 21205 USA Johns Hopkins Med Dept Plast & Reconstruct Surg 601N Caroline St Baltimore MD 21287 USA Johns Hopkins Sch Med Dept Med Div Dermatol 601N Caroline St Baltimore MD 21287 USA Johns Hopkins Univ Dept Med Div Pulm & Crit Care Med Sch Med 1800 Orleans St Baltimore MD 21287 USA

Background: Sweat chloride concentration is used both for CF diagnosis and for tracking CFTR modulator efficacy over time, but the relationship between sweat chloride and lung health is heterogeneous and informed by CFTR genotype. Here, we endeavored to characterize ion transport in eccrine sweat glands (ESGs). Methods: First, ESGs were microdissected from a non-CF skin donor to analyze individual glands. We established primary cultures of ESG cells via conditional reprogramming for functional testing of ion transport by short circuit current measurement and examined cell composition by single-cell RNA-sequencing (scRNA-seq) comparing with whole dissociated ESGs. Secondly, we cultured nasal epithelial (NE) cells and ESGs from two people with CF (pwCF) to assess modulator efficacy. Finally, NEs and ESGs were grown from one person with the CFTR genotype F312del/F508del to explore genotype-phenotype heterogeneity. Results: ESG primary cells from individuals without CF demonstrated robust ENaC and CFTR function. scRNA-seq demonstrated both secretory and ductal ESG markers in cultured ESG cells. In both NEs and ESGs from pwCF homozygous for F508del, minimal baseline CFTR function was observed, and treatment with CFTR modulators significantly enhanced function. Notably, NEs from an individual bearing F312del/F508del exhibited significant baseline CFTR function, whereas ESGs from the same person displayed minimal CFTR function, consistent with observed phenotype. Conclusions: This study has established a novel primary culture technique for ESGs that allows for functional ion transport measurement to assess modulator efficacy and evaluate genotype-phenoytpe heterogeneity. To our knowledge, this is the first reported application of conditional reprogramming and scRNA-seq of microdissected ESGs.

关键词： Eccrine sweat glands conditional reprogramming scRNA-seq Genotype-phenotype Modulator heterogeneity CFTR function

来源：评论

学校读者我要写书评

暂无评论

Insights into the characteristics of primary radioresistant cervical cancer using single-cell transcriptomics

引用

HUMAN CELL 2023年第3期36卷 1135-1146页

作者： Xing, Biyuan Pu, Congli Chen, Yunshang Sheng, Yuhan Zhang, Baofang Cui, Jie Wu, Gang Zhao, Yingchao Huazhong Univ Sci & Technol Union Hosp Tongji Med Coll Canc Ctr Wuhan 430022 Peoples R China Huazhong Univ Sci & Technol Union Hosp Inst Radiat Oncol Tongji Med Coll Wuhan 430022 Peoples R China Chinese Acad Sci Inst Pasteur Shanghai Ctr Biosafety Mega Sci CAS Key Lab Mol Virol & Immunol Shanghai Peoples R China

Radioresistance is a major cause of radiotherapy failure among patients with cervical cancer (CC), the fourth most common cause of cancer mortality in women worldwide. Traditional CC cell lines lose intra-tumoral heterogeneity, posing a challenge for radioresistance research. Meanwhile, conditional reprogramming (CR) maintains intra-tumoral heterogeneity and complexity, as well as the genomic and clinical characteristics of original cells and tissues. Three radioresistant and two radiosensitive primary CC cell lines were developed under CR conditions from patient specimens, and their characteristics were verified via immunofluorescence, growth kinetics, clone forming assay, xenografting, and immunohistochemistry. The CR cell lines had homogenous characteristics with original tumor tissues and maintained radiosensitivity in vitro and in vivo, while also maintaining intra-tumoral heterogeneity according to single-cell RNA sequencing analysis. Upon further investigation, 20.83% of cells in radioresistant CR cell lines aggregated in the G2/M cell cycle phase, which is sensitive to radiation, compared to 38.1% of cells in radiosensitive CR cell lines. This study established three radioresistant and two radiosensitive CC cell lines through CR, which will benefit further research investigating radiosensitivity in CC. Our present study may provide an ideal model for research on development of radioresistance and potential therapeutic targets in CC.

关键词： conditional reprogramming Primary cell lines Single-cell transcriptomics Cervical cancer Radioresistance

来源：评论

学校读者我要写书评

暂无评论

Hormonal regulation of non-cystic fibrosis transmembrane conductance regulator ion channels in the endocervix

F&S SCIENCE

引用

F&S SCIENCE 2023年第2期4卷 163-171页

作者： Roberts, Mackenzie Yao, Shan Wei, Shuhao Jensen, Jeffrey T. Han, Leo Oregon Natl Primate Res Ctr Div Reprod & Dev Sci Portland OR USA Oregon Hlth & Sci Univ Dept Obstet & Gynecol Portland OR USA

Objective: To characterize ion channel expression and localization in the endocervix under different hormonal conditions using a nonhuman primate primary endocervical epithelial cell model. Design: Experimental. Setting: University-based, translational science laboratory. Interventions: We cultured and treated conditionally reprogrammed primary rhesus macaque endocervix cells with estradiol and progesterone and measured gene expression changes for several known ion channel and ion channel regulators of mucus secreting epithelia. Using both rhesus macaque endocervical samples and human samples, we localized channels in the endocervix using immunohistochemistry. Main Outcome Measures: The relative abundance of transcripts was evaluated using real-time polymerase chain reaction. Immunostaining results were evaluated qualitatively. Results: Compared with controls, we found that estradiol increased gene expression for ANO6, NKCC1, CLCA1, and PDE4D. Progesterone down-regulated gene expression for ANO6, SCNN1A, SCNN1B, NKCC1, and PDE4D (P <=.05). Immunohistochemistry confirmed endocervical cell membrane localization of ANO1, ANO6, KCNN4, LRR8CA, and NKCC1. Conclusions: We found several ion channels and ion channel regulators that are hormonally sensitive in the endocervix. These channels, therefore, may play a role in the cyclic fertility changes in the endocervix and could be further investigated as targets for future fertility and contraceptive studies. (Fertil Steril Sci (R) 2023;4:163-71. (c) 2023 by American Society for Reproductive Medicine.)

关键词： Ion channels mucus endocervix hormones conditional reprogramming

来源：评论

学校读者我要写书评

暂无评论

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

请选择保存的检索档案：

请选择收藏分类：

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

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

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：