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Asialoglycoprotein receptor 1 promotes SARS-CoV-2 infection of human normal hepatocytes

Signal Transduction and Targeted Therapy 2024年第3期9卷 1199-1208页

作者： Xinyi Yang Xu Zheng Yuqi Zhu Xiaying Zhao Jun Liu Jiangna Xun Songhua Yuan Jun Chen Hanyu Pan Jinlong Yang Jing Wang Zhimin liang Xiaoting Shen Yue liang Qinru Lin huitong liang Min Li Fei Peng Daru Lu Jianqing Xu Hongzhou Lu Shibo Jiang Ping Zhao Huanzhang Zhu State Key Laboratory of Genetic Engineering and Engineering Research Center of Gene Technology Ministry of EducationInstitute of GeneticsSchool of Life SciencesYiwu Research InstituteFudan UniversityShanghaiChina Department of Microbiology Faculty of Naval MedicineNaval Medical UniversityShanghai 200433China Scientific Research Center Shanghai Public Health Clinical CenterFudan UniversityShanghaiChina Department of Infectious Diseases and Immunology Shanghai Public Health Clinical CenterFudan UniversityShanghaiChina Changzheng Hospital Naval Medical UniversityShanghaiChina Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) Shanghai Institute of Infectious Disease and BiosecuritySchool of Basic Medical SciencesFudan UniversityShanghaiChina Department of Infectious Diseases and Nursing Research Institution National Clinical Research Center for Infectious DiseasesThe Third People’s Hospital of ShenzhenShenzhenGuangdongChina

Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)causes multi-organ damage,which includes hepatic dysfunction,as observed in over 50%of COVID-19 *** I converting enzyme(peptidyl-dipeptidase A)2(ACE2)is the primary receptor for SARS-CoV-2 entry into host cells,and studies have shown the presence of intracellular virus particles in human hepatocytes that express ACE2,but at extremely low ***,we asked if hepatocytes might express receptors other than ACE2 capable of promoting the entry of SARS-CoV-2 into *** address this question,we performed a genome-wide CRISPR-Cas9 activation library screening and found that Asialoglycoprotein receptor 1(ASGR1)promoted SARS-CoV-2 pseudovirus infection of HeLa *** Huh-7 cells,simultaneous knockout of ACE2 and ASGR1 prevented SARS-CoV-2 pseudovirus *** the immortalized THLE-2 hepatocyte cell line and primary hepatic parenchymal cells,both of which barely expressed ACE2,SARSCoV-2 pseudovirus could successfully establish an ***,after treatment with ASGR1 antibody or siRNA targeting ASGR1,the infection rate significantly dropped,suggesting that SARS-CoV-2 pseudovirus infects hepatic parenchymal cells mainly through an ASGR1-dependent *** confirmed that ASGR1 could interact with Spike protein,which depends on receptor binding domain(RBD)and N-terminal domain(NTD).Finally,we also used Immunohistochemistry and electron microscopy to verify that SARS-CoV-2 could infect primary hepatic parenchymal *** inhibiting ASGR1 in primary hepatic parenchymal cells by siRNA,the infection efficiency of the live virus decreased ***,these findings indicate that ASGR1 is a candidate receptor for SARS-CoV-2 that promotes infection of hepatic parenchymal cells.

关键词： ACE2 infection glycoprotein

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一种高活性γδT细胞的体外培养方案

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复旦学报（自然科学版） 2024年第3期63卷 380-388页

作者：林沁汝梁卉彤杨辛毅杨金龙朱豫琪梁玥李敏成逸鹏朱焕章复旦大学生命科学学院上海200433

γδT细胞的体外大规模扩增一般需较长周期,获取高纯度细胞时往往难以保持细胞活性。本文建立了一种无饲养细胞、无外源血清的优化的人γδT细胞体外培养方案。基于唑来膦酸(ZOL)和CD3激活剂从人外周血单个核细胞(PBMC)中扩增γδT细胞... 详细信息

γδT细胞的体外大规模扩增一般需较长周期,获取高纯度细胞时往往难以保持细胞活性。本文建立了一种无饲养细胞、无外源血清的优化的人γδT细胞体外培养方案。基于唑来膦酸(ZOL)和CD3激活剂从人外周血单个核细胞(PBMC)中扩增γδT细胞,在培养第6天得到纯度90%以上的γδT细胞。以CD62L和CD45RA为标记检测扩增产物的记忆分化表型,70%以上为naive T细胞和T记忆干细胞;检测CD69和CD25检测结果表明细胞处于早中期活化状态,PD-1、LAG-3和TIM-3检测结果表明细胞衰竭程度较低。将扩增产物与Jurkat细胞系共孵育,分别通过脱颗粒、IFN-γ分泌和肿瘤细胞杀伤率证实了扩增产物具备较高的Jurkat细胞毒性。使用慢病毒感染扩增产物,证实扩增产物可接受高效的基因转染。本研究提供了可用于过继性细胞移植的高活性γδT细胞,为肿瘤免疫疗法的深入研究及临床应用创造了新的可能。

关键词： γδT细胞唑来膦酸肿瘤免疫

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小分子药物提高慢病毒对人γδT细胞感染效率的研究

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复旦学报（自然科学版） 2024年第4期63卷 492-501页

作者：梁卉彤林沁汝杨辛毅杨金龙朱豫琪梁玥李敏成逸鹏朱焕章复旦大学生命科学学院上海200438

γδT细胞识别靶细胞表面不依赖于主要组织相容性复合体(MHC)分子呈递的抗原,高效杀伤靶细胞且不引起移植物抗宿主病(GvHD),作为细胞平台在同种异体CAR细胞疗法开发中具有良好的应用前景。慢病毒(Lentivirus)载体是CAR-T细胞制备中常用... 详细信息

γδT细胞识别靶细胞表面不依赖于主要组织相容性复合体(MHC)分子呈递的抗原,高效杀伤靶细胞且不引起移植物抗宿主病(GvHD),作为细胞平台在同种异体CAR细胞疗法开发中具有良好的应用前景。慢病毒(Lentivirus)载体是CAR-T细胞制备中常用的基因递送手段,为实现γδT细胞中基于慢病毒载体的高效基因递送,本研究探索能够提高慢病毒滴度以及γδT细胞慢病毒感染效率的小分子药物。结果表明:组蛋白去乙酰化酶抑制剂M344和微管抑制剂Nocodazole可有效提高慢病毒滴度,其中当浓度为0.625μmol/L时,M344可提高慢病毒滴度至2倍以上。TBK1/IKK epsilon抑制剂BX-795可提高γδT细胞中慢病毒感染效率,当感染复数(MOI)为5时,BX-795可将感染效率从3.5%提升至10.7%,且未产生细胞毒性作用。本研究将为γδT细胞中基于慢病毒载体的基因递送优化提供一种新的策略,进一步推动γδT细胞在同种异体CAR细胞研究中的应用。

关键词： γδT细胞慢病毒载体 M344 诺考达唑 BX-795

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Allogeneic gene-edited HIV-specific CAR-T cells secreting PD-1 blocking scFv enhance specific cytotoxic activity against HIV Envt cells in vivo

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Virologica Sinica 2023年第2期38卷 285-295页

作者： Hanyu Pan Xinyi Yang Jing Wang huitong liang Zhengtao Jiang Lin Zhao Yanan Wang Zhiming liang Xiaoting Shen Qinru Lin Yue liang Jinglong Yang Panpan Lu Yuqi Zhu Min Li Pengfei Wang Jianqing Xu Hongzhou Lu Huanzhang Zhu State Key Laboratory of Genetic Engineering And Key Laboratory of Medical Molecular Virology of Ministry of Education/HealthSchool of Life SciencesFudan UniversityShanghai200438China Shanghai Institute of Infectious Disease and Biosecurity School of Life SciencesFudan UniversityShanghai200438China Department of Infectious Disease Key Laboratory of Medical Molecular Virology of Ministry of Education/HealthSchool of Basic Medical Sciences and Shanghai Public Health Clinical CenterFudan UniversityShanghai201508China Scientific Research Center Shanghai Public Health Clinical CenterFudan UniversityShanghai201508China Department of Infectious Diseases and Immunology Shanghai Public Health Clinical CenterFudan UniversityShanghai201508China Department of Infectious Diseases and Nursing Research Institution National Clinical Research Center for Infectious DiseasesThe Third People's Hospital of ShenzhenShenzhen518112China

HIV-specific chimeric antigen receptor(CAR)T-cells have been developed to target HIV-1 infected CD4t T-cells that express HIV Env ***,T cell exhaustion and the patient-specific autologous paradigm of CAR-T cell hurdled clinical ***,we created HIV-specific CAR-T cells using human peripheral blood mononuclear cells and a 3BNC117-E27(3BE)CAR construct that enabled the expression of programmed cell death protein(PD-1)-blocking scFv E27 and the single-chain variable fragment of the HIV-1-specific broadly neutralizing antibody 3BNC117 to target native HIV *** with T cells expressing 3BNC117-CAR alone,3BE CAR-T cells showed greater cytotoxic activity against HIV Envt cells with stronger proliferation capability,higher killing efficiency,and enhanced cytokine secretion in the presence of HIV Env-expressing ***,we manufactured TCR-deficient 3BE CAR-T cells through gene editing and demonstrated that these CAR-T cells could effectively kill HIV Env^(+) cells in vivo without the occurrence of severe graft-versus-host disease(GvHD)in NSG *** data suggest that we have provided a feasible approach to the generation of“off-theshelf”anti-HIV CAR-T cells in combination with PD-1 checkpoint blockade immunotherapy,which can be a powerful therapeutic candidate for the functional cure of HIV.

关键词： Anti-HIV CAR-T PD-1 blocking TCR disruption Cellular immunotherapy

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Neutralization of five SARS-CoV-2 variants of concern by convalescent and BBIBP-CorV vaccinee serum

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Virologica Sinica 2022年第6期37卷 831-841页

作者： Yuqi Zhu Xinyi Yang Jingna Xun Jun Liu Qing Wen Yixiao Lin Xiaoting Shen Jun Chen Songhua Yuan Xiaying Zhao Jing Wang Hanyu Pan Jinlong Yang Zhiming liang Yue liang Qinru Lin huitong liang Min Li Jianping Liu Yinzhong Shen Xiaoyan Zhang Pengfei Wang Daru Lu Chunhua Yin Jianqing Xu Shibo Jiang Hongzhou Lu Huanzhang Zhu State Key Laboratory of Genetic Engineering and Engineering Research Center of Gene Technology Ministry of EducationInstitute of GeneticsSchool of Life SciencesFudan UniversityShanghai200438China Scientific Research Center Shanghai Public Health Clinical CenterFudan UniversityShanghai201508China Fubio(Suzhou)Biomedical Technology Co. LtdSuzou215300China Department of Infectious Diseases and Immunology Shanghai Public Health Clinical CenterFudan UniversityShanghai201508China Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) School of Basic Medical SciencesFudan UniversityShanghai200032China Department of Infectious Diseases and Nursing Research Institution National Clinical Research Center for Infectious DiseasesThe Third People's Hospital of ShenzhenShenzhen518112China

The prevalence of SARS-CoV-2 variants of concern(VOCs) is still escalating throughout the world. However, the level of neutralization of the inactivated viral vaccine recipients’ sera and convalescent sera against all VOCs,including B.1.1.7(Alpha), B.1.351(Beta), P.1(Gamma), B.1.617.2(Delta), and B.1.1.529(Omicron) remains to be lack of comparative analysis. Therefore, we constructed pseudoviruses of five VOCs using a lentiviral-based system and analyzed their viral infectivity and neutralization resistance to convalescent and BBIBP-CorV vaccinee serum at different times. Our results show that, compared with the wild-type strain(WT), five VOC pseudoviruses showed higher infection, of which B.1.617.2 and B.1.1.529 variant pseudoviruses exhibited higher infection rates than wild-type or other VOC strains, respectively. Sera from 10 vaccinated individuals at the 1, 3and 5-month post second dose or from 10 convalescent at 14 and 200 days after discharge retained neutralizing activity against all strains but exhibited decreased neutralization activity significantly against the five VOC variant pseudoviruses over time compared to WT. Notably, 100%(30/30) of the vaccinee serum samples showed more than a 2.5-fold reduction in neutralizing activity against B.1.1.529, and 90%(18/20) of the convalescent serum samples showed more than 2.5-fold reduction in neutralization against B.1.1.529. These findings demonstrate the reduced protection against the VOCs in vaccinated and convalescent individuals over time, indicating that it is necessary to have a booster shot and develop new vaccines capable of eliciting broad neutralization antibodies.

关键词： SARS-CoV-2 Variants of concern(VOCs) Convalescent BBIBP-CorV vaccine Neutralization

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基于多尺度注意力特征融合的双目深度估计算法

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激光与光电子学进展 2022年第18期59卷 259-267页

作者：杨蕙同雷亮林永春广东工业大学物理与光电工程学院广东广州510006

针对目前端到端的立体匹配算法在具有挑战性的复杂场景中出现的误匹配现象,提出一种多尺度注意力特征融合立体匹配算法(MGNet)。设计了一个轻量级的组相关注意力模块,该模块采用组相关融合单元来有效地结合空间注意机制与通道注意力机制... 详细信息

针对目前端到端的立体匹配算法在具有挑战性的复杂场景中出现的误匹配现象,提出一种多尺度注意力特征融合立体匹配算法(MGNet)。设计了一个轻量级的组相关注意力模块,该模块采用组相关融合单元来有效地结合空间注意机制与通道注意力机制,同时捕获丰富的全局上下文信息和远距离通道依赖关系。设计了多尺度卷积全局注意力模块,它能够在多个尺度下处理局部和全局信息,在全局特征处理阶段引入非局部操作,可以同时捕获多尺度上下文与全局上下文,提供丰富的语义信息。在代价聚合阶段引入通道注意力,抑制具有歧义的匹配信息,提取有区别性的特征。使用三大数据集评估了所提算法的有效性,由实验结果可知,所提算法在薄结构、反射区域、弱纹理、重复纹理等复杂场景均表现优异。

关键词：深度估计立体匹配深度学习注意力机制

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Hexagonal boron nitride nanosheet for effective ambient N2 fixation to NH3

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Nano Research 2019年第4期12卷 919-924页

作者： Ya Zhang huitong Du Yongjun Ma Lei Ji Haoran Guo Ziqi Tian Hongyu Chen Hong Huang Guanwei Cui Abdullah M. Asiri Fengli Qu liang Chen Xuping Sun Institute of Fundamental and Frontier Sciences University of Electronic Science and Technology of China Chengdu 610054 China College of Chemistry and Chemical Engineering Qufu Normal University Qufu 273165 China Analytical and Test Center Southwest University of Science and Technology Mianyang 621010 China Ningbo Institute of Materials Technology and Engineering Chinese Academy of Sciences Ningbo 315201 China College of Chemistry Chemical Engineering and Materials Science Shandong Normal University Jinan 250014 China Chemistry Department Faculty of Science & Center of Excellence for Advanced Materials Research King Abdulaziz University P.O. Box 80203 Jeddah 21589 Saudi Arabia

Industrial production of NH3 from N2 and H2 significantly relies on Haber-Bosch process,which suffers from high energy consume and CO2 *** a sustainable and environmentally-benign alternative process,electrochemical artificial N2 fixation at ambient conditions,however,is highly required efficient *** this study,we demonstrate that hexagonal boron nitride nanosheet (h-BNNS) is able to electrochemically catalyze N2 to *** acidic solution,h-BNNS catalyst attains a high NH3 formation rate of 22.4 μg·h-1·*** a high Faradic efficiency of 4.7% at-0.75 V *** hydrogen electrode,with excellent stability and *** functional theory calculations reveal that unsaturated boron at the edge site can activate inert N2 molecule and significantly reduce the energy barrier for NH3 fonmation.

关键词： boron nitride nanosheet N2 reduction reaction NH3 electrosynthesis ambient conditions density functional theory

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线上供应链金融平台客户信息安全风险自动评估系统设计

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自动化技术与应用 2024年第6期43卷 116-119页

作者：朱辉袁亮张洁孙琴国网数字科技控股有限公司国网雄安金融科技集团有限公司北京100053 国网汇通金财(北京)信息科技有限公司北京100053

风险因素级别估算过程中,安全风险自动评估权值的绝对误差增加,评估结果可信度降低,由此,提出一种线上供应链金融平台客户信息安全风险自动评估系统。通过分析结果向系统管理员提供周密可靠的安全性分析报告,同时结合评估数据库进行系... 详细信息

风险因素级别估算过程中,安全风险自动评估权值的绝对误差增加,评估结果可信度降低,由此,提出一种线上供应链金融平台客户信息安全风险自动评估系统。通过分析结果向系统管理员提供周密可靠的安全性分析报告,同时结合评估数据库进行系统硬件部分设计。将人工神经网络应用到客户信息安全风险评估中,估算风险因素的级别,建立自动评估模型,模型在经过训练后能够有效实现线上供应链金融平台的客户信息安全风险自动评估。仿真实验结果表明,所设计系统能够有效减少绝对误差,提升评估结果的可信度。

关键词：线上供应链金融平台客户信息安全风险自动评估系统设计

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基于改进残差网络的多视图焊点缺陷检测

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焊接学报 2022年第3期43卷 56-62,I0006页

作者：邓智超颜润明杨蕙同陈浩林赖锦祥雷亮广东工业大学广州510006

基于卷积神经网络的单视图检测不能有效识别三维形状的缺陷目标,导致在实际应用中,往往是通过只检测某一最具代表性的视图或者依次检测每个面来实现低精度的检测要求,这带来了较大的时间成本和使用限制.针对这一问题,提出了改进的残差网... 详细信息

基于卷积神经网络的单视图检测不能有效识别三维形状的缺陷目标,导致在实际应用中,往往是通过只检测某一最具代表性的视图或者依次检测每个面来实现低精度的检测要求,这带来了较大的时间成本和使用限制.针对这一问题,提出了改进的残差网络(Res Net),并将其应用于三维形状的焊点缺陷检测.该模型首先会一次性获取焊点的所有视图图像,再通过特征聚合和自适应学习模块,最终获得检测结果.多视图焊点数据集来自高频电感元件,在所提出方法的识别精度达到了99.48%.结果表明,改进的残差网络在同等网络层数的情况下有效提升了图像识别精度;对比单视图检测,多视图检测结构仅以较少的时间代价获得了较大的精度提升,能有效完成实际工业生产中的三维形状缺陷目标的检测任务.

关键词：缺陷检测多视图特征融合焊点卷积神经网络

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Plant functional trait diversity and structural diversity co-underpin ecosystem multifunctionality in subtropical forests

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Forest Ecosystems 2023年第2期10卷 153-161页

作者： Shuai Ouyang Mengmeng Gou Pifeng Lei Yue Liu liang Chen Xiangwen Deng Zhonghui Zhao Yelin Zeng Yanting Hu Changhui Peng Wenhua Xiang Faculty of Life Science and Technology Central South University of Forestry and TechnologyChangsha410004China huitong National Station for Scientific Observation and Research of Chinese Fir Plantation Ecosystems in Hunan Province Huitong438107China Ecology and Nature Conservation Institute Chinese Academy of ForestryBeijing100101China Institute of Environment Sciences Department of Biological SciencesUniversity of Quebec at MontrealMontrealQCH3C 3P8Canada

Tree species diversity is assumed to be an important component in managing forest ecosystems because of effects on multiple functions or ecosystem ***,the importance of tree diversity in determining multifunctionality in structurally complex subtropical forests relative to other regulators(e.g.,soil microbial diversity,stand structure,and environmental conditions)remains *** this study,effects of aboveground(species richness and functional and structural diversity)and belowground(bacterial and fungal diversity)biodiversity,functional composition(community-weighted means of species traits),stand structure(diameter at breast height and stand density),and soil factors(pH and bulk density)on multifunctionality(including biomass production,carbon stock,and nutrient cycling)were examined along a tree diversity gradient in subtropical *** community-weighted mean of tree maximum height was the best predictor of ecosystem *** diversity explained a higher proportion of the variation in multifunctionality than that of species richness and fungal *** structure-played an important role in modulating the effects of tree diversity on *** work highlights that species composition and maximizing forest structural complexity are effective strategies to increase forest multifunctionality while also conserving biodiversity in the management of multifunctional forests under global environmental changes.

关键词： Abiotic and biotic factors Biodiversity Functional composition Functional traits Soil microbial diversity Stand structure

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