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李大力(华东师範大学生命科学学院研究员)

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李大力(华东师範大学生命科学学院研究员)

李大力(华东师範大学生命科学学院研究员)

2004年10月至2007年5月在美国德州农工大学健康科学中心的生物科学与技术研究所从事研究助理的工作，作为联合培的博士生于2007年6月回国获得湖南师範大学遗传学博士学位。毕业后受聘于华东师範大学生命科学学院生命医学研究所从事教学与科研工作。主要负责基因敲除与转基因小鼠动物中心的建设和G蛋白偶联受体相关的科学研究。已发表科研论文14篇，其中有10篇发表在《PNAS》、《ONCONGENE》和《ENDOCRINOLOGY》等SCI期刊上，参编专着一部，另有3篇第一作者论文和2篇合作研究论文已投稿。主持国家自然科学基金青年基金项目一项（2009－2011），作为主要成员参与上海市科委研发公共服务平台“细胞信号网路研究技术平台”建设（2007-2010）。

基本介绍

中文名：李大力外文名：Dali Li国籍：中国职业：华东师範大学生命科学学院研究员毕业院校：湖南师範大学

个人简历

李大力，华东师範大学生命科学学院研究员。2016年10月10日，来自中国科学院、北京大学、浙江大学、上海交通大学、华东师範大学、哈尔滨工业大学、温州医科大学等科研院所的13位课题组负责人实名公开表示，无法重複韩春雨2016年5月2日发表在《自然-生物技术》上有关NgAgo的实验。发表公开声明的13位学者分别是：北京大学生命科学学院教授魏文胜，北京大学生命科学学院研究员孙育杰，北京大学分子医学研究所教授熊敬维，中科院动物研究所研究员王皓毅、李伟，中科院生物物理研究所研究员王晓群，中科院生物化学与细胞生物学研究所研究员李劲松，中科院上海生科院神经科学研究所研究员杨辉，浙江大学生命科学研究院教授王立铭，上海交通大学教授吴强，华东师範大学生命科学学院研究员李大力，哈尔滨工业大学教授黄志伟，温州医科大学教授谷峰。

教育经历

2001.9-2007.6 博士研究生 湖南师範大学生命科学学院2004.9-2007.6 联合培养 美国德州农工大学健康科学中心1997.9-2001.7 本科生 湖南师範大学生命科学学院

个人荣誉

教育部“长江学者奖励计画”青年学者上海市普陀区“拔尖人才”

学术兼职

　　Scientific Reports编委Nature Biotechnology, Nature Protocols, Cell Research等杂誌审稿人

研究方向

1、动物模型构建新技术开发基因修饰动物模型是我们的主要研究对象和研究手段，课题组重点发展、最佳化基因修饰动物模型构建技术。2013年初，我们建立了基于TALEN的基因敲除小鼠平台（Nucleic Acids Research， 2013），是世界上最早利用该技术构建基因敲除小鼠的实验室之一。在此基础上，利用TALEN技术构建了基因敲除大鼠的技术体系，开展以基因修饰小鼠和大鼠等两种模式动物的基础研究（SCI CHINA LIFE SCI，2016）。 2013年8月，课题组报导了利用CRISPR/Cas9技术构建基因修饰大鼠和小鼠模型的方法，在世界上首次实现同时构建多基因敲除大鼠（Nature Biotechnology， 2013），并应邀撰写了基因修饰大鼠构建的详细操作步骤（Nature Protocols，2014；Methods Enzymol.2014）。通过该方法将基因修饰大鼠和小鼠的构建时间缩短到5周左右，极大的提高了效率。利用重组Cas9蛋白替代mRNA进行胚胎注射，降低脱靶活性而提升了同源重组效率，实现大片段DNA的删除和定点基因插入，丰富了基因编辑技术在发育生物学工具小鼠和大鼠构建的套用多样性（Scientific Reports，2015）。作为合作者，我们构建的动物模型得到广泛套用，相关论文发表在Nature Medicine （2016），Molecular Cell（2015）和Nature Communications（2015）等杂誌。到目前为止，Cas9的脱靶问题和重组效率低一直是制约该技术进一步发展和套用的主要原因，本课题组也将重点研究如何提高同源重组效率，促进技术发展，採用非天然胺基酸定点整合的方法在哺乳动物中率先实现遗传密码扩充，证明哺乳动物可以耐受遗传密码的人为改变（Cell Research，2017）。2、基因编辑技术在基因治疗中的套用遗传疾病的治疗往往需要长期用药，很难达到根治的目的，极大的影响患者的生活质量，给病人和家庭带来沉重负担。基因治疗有望在细胞中重新表达受损基因而获得长期疗效，但在实际操作中受到非常多的限制。基因编辑技术出现后，能够对基因组DNA进行精确操作，极大降低了随机整合的风险，有望修复基因实现长期表达，为基因治疗的相关研究注入了新的活力。课题组利用CRISPR/Cas9基因编辑技术模拟在病人中新发现的凝血因子9突变，构建点突变小鼠，建立了新的B型血友病动物模型。利用定点修复成年B型血友病小鼠模型中F9基因突变策略，开展缓解凝血功能障碍的研究。通过尾静脉注射将Cas9/sgRNA及对应的同源重组修复模板导入成年突变小鼠体内，成功地在0.56-2.84%的肝细胞中完全修复F9基因突变，显着缩短活化部分凝血活酶时间（aPTT），极大提高了断尾失血实验中血友病小鼠的存活率。在国际上率先证明了通过Cas9基因编辑技术原位修复F9基因突变治疗B型血友病的可行性，也为基因编辑技术套用于单基因遗传病的治疗提供了有力的实验支持（EMBO Molecular Medicine，2016）。课题组将在此基础上，利用AAV病毒体系导入Cas9体系，尝试通过基因编辑治疗其他遗传疾病的可行性；通过增强基因导入效率、提高重组修复等方法获得更高效的基因修复效率，达到更好的治疗效果；建立造血干细胞等可移植细胞基因编辑技术体系，开展离体基因编辑与细胞治疗的套用研究。3.成体干细胞自我更新和分化机制研究肠上皮组织是动物体中更新最快的组织之一，是理想的研究成体干细胞分化以及组织损伤修复的模型。同时，由于多种原因导致的肠上皮的损伤也与肠炎有着密切的联繫。肠干细胞如何受到外界刺激引起内在信号转导通路的变化最终表现为其自我更新或者终末分化，这是一个非常複杂而又尚待解决的重要科学问题。通过研究Lgr4/Gpr48基因敲除小鼠小肠上皮细胞分化与修复的缺陷，证明了Lgr4介导的R-spondin信号通过直接调节Wnt/beta-Catenin信号通路在肠干细胞稳态维持和再生中的重要功能（JBC， 2013， paper of the week）。以此研究为契机迅速建立了肠干细胞离体3D培养体系（organoid culture）和检测方法，并利用该体外培养模型，筛选可能调节肠干细胞增殖、分化的胞外信号分子。通过基因编辑技术，建立负责基因修饰小鼠模型，研究相关信号通路在肠干细胞自我更新和分化的分子机制和生理病理表型。

主持课题

    青春发育关键基因KiSS1表达调控研究（国家自然科学基金No.30800627 2009-2011 ）核仁蛋白PAK1IP1 调节核糖体生物合成的功能及机理研究 （国家自然科学基金 No.31171318 2012-2015）七次跨膜受体Lgr4 及其配体家族在卵巢功能维持和雌性不育中的功能研究 (国家自然科学基金No.31371455 2014-2017 )构建PAH基因新突变小鼠模型开展苯丙酮尿症基因治疗研究 (国家自然科学基金 No. 81670470 2017-2020)重症免疫缺陷型大鼠品系的构建以及基因编辑技术的最佳化研究 (上海市创新课题 No.14140900300, 2014-2017）

发表论文

5篇代表论文
    Li D*, Qiu Z, Shao Y, Chen Y, Guan Y, Liu M, Li Y, Gao N, Wang L, Lu X, Zhao Y*, Liu M*. Heritable gene targeting in the mouse and rat using a CRISPR-Cas system.Nature Biotechnology. 2013 ;31(8):681-3Guan Y, Ma L, Li Q, Sun Z, Ma Li, Wu L, Wang L, Zeng L, Shao Y, Chen Y, Ma N, Lu W, Hu K, Han H, Yu Y, Huang Y, Liu M*,Li D*. CRISPR/Cas9 mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse.EMBO Mol Med. 2016 May 2;8(5):477-88News&Views,Nguyen TH, Anegon I. Successful correction of hemophilia by CRISPR/Cas9 genome editing in vivo: delivery vector and immune responses are the key to success.EMBO Mol Med. 2016 May 2;8(5):439-41.) （杂誌同期点评）Chen Y, Ma J, Lu W, Tian M, Thauvin M, Yuan C, Volovitch M, Wang Q, Holst J, Liu M, Vriz S, Ye S*, Wang L*,Li D*. Heritable expansion of the genetic code in mouse and zebrafish.Cell Res. 2016 Dec 9.Editor’s Choice, Valda Vinson, Expanding the genetic code in vertebrates,Science2017:Vol. 355, Issue 6320, pp. 36 (Science杂誌编辑选为合成生物学方面当月亮点工作)Shao Y, Guan Y, Wang L, Qiu Z, Liu M, Chen Y, Wu L, Li Y, Ma X, Liu M*,Li D*. Genome editing in the rat using CRISPR-Cas and injection of RNAs into one-cell embryos.Nature Protocols. 2014 (10):2493-512Qiu Z, Liu M, Chen Z, Shao Y, Pan H, Wei G, Yu C, Zhang L, Li X, Wang P, Fan HY, Du B, Liu B, Liu M*,Li D*. High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases.Nucleic Acids Res.2013 Jun;41(11):e120
论文列表：2017：
    Zhang L, Li L, Gao G, Wei G, Zheng Y, Wang C, Gao N, Zhao Y, Deng J, Chen H, Sun J,Li D*, Zhang X*, Liu M*. Elevation of GPRC5A expression in colorectal cancer promotes tumor progression through VNN-1 induced oxidative stress.Int J Cancer.2017 Mar 17. doi: 10.1002/ijc.30698.Lu J, Shao Y, Qin X, Liu D, Chen A,Li D*, Liu M*, Wang X*. CRISPR knockout rat cytochrome P450 3A1/2 model for advancing drug metabolism and pharmacokinetics research.Sci Rep.2017 Feb 20;7:42922.Chen Y, Ma J, Lu W, Tian M, Thauvin M, Yuan C, Volovitch M, Wang Q, Holst J, Liu M, Vriz S, Ye S*, Wang L*,Li D*. Heritable expansion of the genetic code in mouse and zebrafish.Cell Res. 2017 Feb;27(2):294-297.Chen R, Zhang Q, Duan X, York P, Chen GD, Yin P, Zhu H, Xu M, Chen P, Wu Q, Li D, Samarut J, Xu G, Zhang P, Cao X, Li J, Wong J. The 5-hydroxymethylcytosine (5hmC) reader Uhrf2 is required for normal levels of 5hmC in mouse adult brain and spatial learning and memory.J Biol Chem.2017 Jan 23.
2016：
    Wei Y, Chen Y, Qiu Y, Zhao H, Liu G, Zhang Y, Meng Q, Wu G, Chen Y, Cai X, Wang H, Ying H, Zhou B, Liu M,Li D, Ding Q. Prevention of Muscle Wasting by CRISPR/Cas9-mediated Disruption of Myostatin In Vivo.Mol Ther.2016 Nov;24(11):1889-1891.You P, Hu H, Chen Y, Zhao Y, Yang Y, Wang T, Xing R, Shao Y, Zhang W,Li D*, Chen H*, Liu M*. Effects of Melanocortin 3 and 4 Receptor Deficiency on Energy Homeostasis in Rats.Sci Rep.2016 Oct 7;6:34938.Chen Y, Liu X, Zhang Y, Wang H, Ying H, Liu M,Li D, Lui KO, Ding Q. A Self-restricted CRISPR System to Reduce Off-target Effects.Mol Ther. 2016 Sep;24(9):1508-10.Zhang L, Shao Y, Li L, Tian F, Cen J, Chen X, Hu D, Zhou Y, Xie W, Zheng Y, Ji Y, Liu M,Li D*, Hui L*. Efficient liver repopulation of transplanted hepatocyte prevents cirrhosis in a rat model of hereditary tyrosinemia type I.Sci Rep. 2016 Aug 11;6:31460Chen Y, Lu W, Gao N, Long Y, Shao Y, Liu MZ, Chen H, Ye S, Ma X, Liu M*,Li D*.Generation of obese rat model by transcription activator-like effector nucleases targeting the leptin receptor gene.Sci China Life Sci.2016 May 25.Luo J, Yang Z, Ma Y, Yue Z, Lin H, Qu G, Huang J, Dai W, Li C, Zheng C, Xu L, Chen H, Wang J,Li D, Siwko S, Penninger JM, Ning G, Xiao J, Liu M. LGR4 is a receptor for RANKL and negatively regulates osteoclast differentiation and bone resorption.Nat Med. 2016 May;22(5):539-46Zhang Y, Zeng Z, Zhao J, Li D, Liu M, Wang X. Measurement of Rhodamine 123 in Three-Dimensional Organoids: A Novel Model for P-Glycoprotein Inhibitor Screening.Basic Clin Pharmacol Toxicol. 2016 Apr 6.Guan Y, Ma L, Li Q, Sun Z, Ma Li, Wu L, Wang L, Zeng L, Shao Y, Chen Y, Ma N, Lu W, Hu K, Han H, Yu Y, Huang Y, Liu M*,Li D*.CRISPR/Cas9 mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse.EMBO MOL MED. 2016 May 2;8(5):477-88.Wang X*, Tang Y, Lu J, Shao Y, Qin X, Li Y, Wang L,Li D*,Liu M*. Characterization of novel cytochrome P450 2E1 knockout rat model generated by CRISPR/Cas9.Biochem Pharmacol. 2016 Apr 1;105:80-90.Liu XM, Zhang YP, Ji SY, Li BT, Tian X, Li D, Tong C, Fan HY. Mitoguardin-1 and -2 promote maturation and the developmental potential of mouse oocytes by maintaining mitochondrial dynamics and functions.Oncotarget. 2016 Jan 12;7(2):1155-67.
2015：
    Wang L, Shao Y, Guan Y, Li L, Wu L, Chen F, Liu M, Chen H, Ma Y, Ma X, Liu M*,Li D*. Large genomic fragment deletion and functional gene cassette knock-in via Cas9 protein mediated genome editing in one-cell rodent embryos.Sci Rep. 2015 Dec 1;5:17517Bai M, Li Q, Shao Y, Huang Y,Li D*, Ma Y*. Generation of site-specific mutant mice using the CRISPR/Cas9 system.Yi Chuan. 2015 Oct;37(10):1029-35.Guan Y, Zhang L, Li X, Zhang X, Liu S, Gao N, Li L, Gao G, Wei G, Chen Z, Zheng Y, Ma X, Siwko S, Chen JL, Liu M*,Li D*. Repression of Mammalian Target of Rapamycin Complex 1 Inhibits Intestinal Regeneration in Acute Inflammatory Bowel Disease Models.J Immunol. 2015 May 29. pii: 1303356Liu XM, Zhang YP, Ji SY, Li BT, Tian X,Li D, Tong C, Fan HY. Mitoguardin-1 and -2 promote maturation and the developmental potential of mouse oocytes by maintaining mitochondrial dynamics and functions.Oncotarget. 2015 doi: 10.18632/oncotarget.6713Deng L, Jiang C, Chen L, Jin J, Wei J, Zhao L, Chen M, Pan W, Xu Y, Chu H, Wang X, Ge X,Li D, Liao L, Liu M, Li L, Wang P. The Ubiquitination of RagA GTPase by RNF152 Negatively Regulates mTORC1 Activation.Mol Cell. 2015 Jun 4;58(5):804-18Fan G, Sun L, Shan P, Zhang X, Huan J, Zhang X,Li D, Wang T, Wei T, Zhang X, Gu X, Yao L, Xuan Y, Hou Z, Cui Y, Cao L, Li X, Zhang S, and Wang C. Loss of KLF14 triggers centrosome amplification and tumorigenesis.Nat Commun2015 Oct 6;6:8450Fang P, Xu W,Li D, Zhao X, Dai J, Wang Z, Yan X, Qin M, Zhang Y, Xu C, Wang L, Qiao Z. A novel acrosomal protein, IQCF1, involved in sperm capacitation and the acrosome reaction.Andrology. 2015 Mar;3(2):332-44.
2014：
    Guan Y, Shao Y,Li D*，Liu M*. Generation of Site-Specific Mutations in the Rat Genome Via CRISPR/Cas9.Methods Enzymol.2014 ;546:297-317Shao Y, Guan Y, Wang L, Qiu Z, Liu M, Chen Y, Wu L, Li Y, Ma X, Liu M*,Li D*. Genome editing in the rat using CRISPR-Cas and injection of RNAs into one-cell embryos.Nature Protocols. 2014 (10):2493-512 （IF，9.67）Cui H, Wang Y, Huang H, Yu W, Bai M, Zhang L, Bryan BA, Wang Y, Luo J,Li D*, Ma Y*, Liu M*. GPR126 Regulates Developmental and Pathological Angiogenesis through Modulation of VEGFR2 Signaling.J Biol Chem.2014 Sep 12. pii: jbc.M114.571000 （IF，4.57）Guan X, Duan Y, Zeng Q, Pan H, Qian Y,Li D*, Cao X*, Liu M*. Lgr4 Deficiency Induces Ataxia-like Phenotype in Mice and Impairs Long-term Depression at Cerebellar Parallel fiber-Purkinje Cell Synapses.J Biol Chem.2014 Sep 19;289(38):26492-504. （IF，4.57）Pan H, Cui H, Liu S, Qian Y, Wu H, Li L, Guan Y, Guan X, Zhang L, Fan HY, Ma Y, Li R, Liu M*,Li D*. Lgr4 gene regulates Corpus Luteum Maturation through Modulation of the WNT mediated EGFR-ERK Signaling Pathway.Endocrinology. 2014 May 30:en20132183. （IF，4.50）Yi T, Weng J, Siwko S, Luo J,Li D, Liu M. LGR4/GPR48 inactivation leads to aniridia-genitourinary anomalies-mental retardation syndrome defects.J Biol Chem.2014 Mar 28;289(13):8767-80
2013：
    Li D*, Qiu Z, Shao Y, Chen Y, Guan Y, Liu M, Li Y, Gao N, Wang L, Lu X, Zhao Y*, Liu M*. Heritable gene targeting in the mouse and rat using a CRISPR-Cas system.Nature Biotechnol.2013 ;31(8):681-3. （IF，41.5）Qiu Z, Liu M, Chen Z, Shao Y, Pan H, Wei G, Yu C, Zhang L, Li X, Wang P, Fan HY, Du B, Liu B, Liu M,Li D*. High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases.Nucleic Acids Res.2013 Jun;41(11):e120. （IF，9.11）Liu S, Qian Y, Li L, Wei G, Guan Y, Pan H, Guan X, Zhang L, Lu X, Zhao Y, Liu M*,Li D*.Lgr4 deficiency increases susceptibility and severity of dextran sodium sulphate induced inflammatory bowel disease in mice.J Biol Chem. 2013 288: 8794-8803. [paper of the week] （IF，4.57）Qian Y, Liu S, Guan Y, Pan H, Guan X, Qiu Z, Li L, Gao N, Zhao Y, Li X, Lu Y, Liu M*,Li D.* Lgr4 mediated Wnt/β-catenin signaling in peritubular myoid cells is essential for spermatogenesis.Development.2013;140(8):1751-61. （IF，6.46）Luo W, Rodriguez M, Valdez JM, Zhu X, Tan K,Li D, Siwko S, Xin L, Liu M. Lgr4 is a Key Regulator of Prostate Development and Prostate Stem Cell Differentiation.Stem Cells.2013;31(11):2492-505. （IF，6.52）Wang Y, Dong J,Li D, Lai L, Siwko S, Li Y, Liu M. Lgr4 regulates mammary gland development and stem cell activity through the pluripotency transcription factor Sox2.Stem Cells. 2013;31(9):1921-31. （IF，6.52）Du B, Luo W, Li R, Tan B, Han H, Lu X,Li D, Qian M, Zhang D, Zhao Y, Liu M. Lgr4/Gpr48 negatively regulates TLR2/4-associated pattern recognition and innate immunity by targeting CD14 expression.J Biol Chem.2013; 288(21):15131-41. （IF，4.57）
2013年以前：
    Yu W, Qiu Z, Gao N, Wang L, Cui H, Qian Y, Jiang L, Luo J, Yi Z, Lu H,Li D*, and Liu M* PAK1IP1, a ribosomal stress-induced nucleolar protein, regulates cell proliferation via the p53-MDM2 loop,Nucleic Acids Res2010 39(6):2234-48. （IF，9.11）Yang Z, Li C, Wang X, Zhai C, Yi Z, Wang L, Liu B, Du B, Wu H, Guo X, Liu M,Li D*, Luo J*. Dauricine induces apoptosis, inhibits proliferation and invasion through inhibiting NF-kappaB signaling pathway in colon cancer cells.J Cell Physiol. 2010 Oct;225(1):266-75. （IF，3.84）Li, D., Yu, W. and Liu, M. (2009) Regulation of KiSS1 gene expression.Peptides, 30, 130-138. （IF，2.62）Li, D*., Niu, Z., Yu, W., Qian, Y., Wang, Q., Li, Q., Yi, Z., Luo, J., Wu, X., Wang, Y. Schwartz RJ, Liu M. (2009) SMYD1, the myogenic activator, is a direct target of serum response factor and myogenin.Nucleic Acids Res, 37, 7059-7071 . （IF，9.11）Li, D., Mitchell, D., Luo, J., Yi, Z., Cho, S.G., Guo, J., Li, X., Ning, G., Wu, X. and Liu, M. (2007) Estrogen regulates KiSS1 gene expression through estrogen receptor alpha and SP protein complexes.Endocrinology,148, 4821-4828. （IF，4.50）Cho SG,Li D, Tan K, Siwko SK, Liu M. KiSS1 and its G-protein-coupled receptor GPR54 in cancer development and metastasis.Cancer Metastasis Rev. 2012；31(3-4):585-91. （IF，7.24）Dai F, Chen Y, Song Y, Huang L, Zhai D, Dong Y, Lai L, Zhang T,Li D, Pang X, Liu M, Yi Z. A natural small molecule harmine inhibits angiogenesis and suppresses tumour growth through activation of p53 in endothelial cells.PLoS One. 2012;7(12):e52162 （IF，3.23）Cho SG, Wang Y, Rodriguez M, Tan K, Zhang W, Luo J,Li D, Liu M Haploinsufficiency in the prometastasis Kiss1 receptor Gpr54 delays breast tumor initiation, progression, and lung metastasis.Cancer Res. 2011 15;71(20):6535-46. （IF，9.33）Dong Y, Lu B, Zhang X, Zhang J, Lai L,Li D, Wu Y, Song Y, Luo J, Pang X, Yi Z, Liu M. Cucurbitacin E, a Tetracyclic Triterpenes Compound from Chinese Medicine, Inhibits Tumor Angiogenesis through VEGFR2 Mediated Jak2/STAT3 Signaling Pathway.Carcinogenesis.2010 Aug 23Yi, T., Tan, K., Cho, S.G., Wang, Y., Luo, J., Zhang, W.,Li, D. and Liu, M. Regulation of embryonic kidney branching morphogenesis and glomerular development by KISS1 receptor (GPR54) through NFAT2 and SP1 mediated bmp7 expression.J Biol Chem. 285(23):17811-20Li, X.Y., Lu, Y., Sun, H.Y., Wang, J.Q., Yang, J., Zhang, H.J., Fan, N.G., Xu, J., Jiang, J.J., Liu, R.Y.,Li,D.et al.(2010) G protein-coupled receptor 48 upregulates estrogen receptor alpha expression via cAMP/PKA signaling in the male reproductive tract.Development, 137, 151-157.Li, C., Yang, Z., Zhai, C., Qiu, W.,Li, D., Yi, Z., Wang, L., Tang, J., Qian, M., Luo, J.et al.Maslinic acid potentiates the anti-tumor activity of tumor necrosis factor alpha by inhibiting NF-kappaB signaling pathway.Mol Cancer, 9, 73Yi, Z.F., Cho, S.G., Zhao, H., Wu, Y.Y., Luo, J.,Li, D., Yi, T., Xu, X., Wu, Z. and Liu, M. 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