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Gene-knockout by iSTOP enables rapid reproductive disease modeling and phenotyping in germ cells of the founder generation

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Science China(life Sciences) 2024年第5期67卷 1035-1050页

作者： Yaling Wang Jingwen Chen Xueying Huang Bangguo Wu Peng Dai Feng Zhang jinsong li lingbo Wang State Key Laboratory of Genetic Engineering Shanghai Key Laboratory of Metabolic Remodeling and HealthInstitute of Metabolism and Integrative BiologyFudan UniversityShanghai 200438China Institute of Reproduction and Development Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China NHC Key Lab of Reproduction Regulation(Shanghai Institute for Biomedical and Pharmaceutical Technologies) School of PharmacyFudan UniversityShanghai 200433China Shanghai Key Laboratoryof Maternal and Fetal Medicine Clinical and Translational Research Center of Shanghai First Maternity and Infant HospitalFrontier Science Center for Stem Cell ResearchSchoolof Life Sciences and TechnologyTongji UniversityShanghai 200092China State Key Laboratory of Cell Biology CAS Center for Excellence in Molecular Cell ScienceShanghai Institute of Biochemistry and Cell BiologyChinese Academy of SciencesShanghai 200031China

Cytosine base editing achieves C·G-to-T·A substitutions and can convert four codons(CAA/CAG/CGA/TGG)into STOP-codons(induction of STOP-codons,iSTOP)to knock out genes with reduced *** enables direct phenotyping in founders’somatic cells,but it remains unknown whether this works in founders’germ cells so as to rapidly reveal novel genes for ***,we initially establish that iSTOP in mouse zygotes enables functional characterization of known genes in founders’germ cells:Cfap43-iSTOP male founders manifest expected sperm features resembling human“multiple morphological abnormalities of the flagella”syndrome(i.e.,MMAF-like features),while oocytes of Zp3-iSTOP female founders have no zona *** further illustrate iSTOP’s utility for dissecting the functions of unknown genes with Ccdc183,observing MMAF-like features and male infertility in Ccdc183-iSTOP founders,phenotypes concordant with those of Ccdc183-KO *** ultimately establish that CCDC183 is essential for sperm morphogenesis through regulating the assembly of outer dynein arms and participating in the intra-flagellar *** study demonstrates iSTOP as an efficient tool for direct reproductive disease modeling and phenotyping in germ cells of the founder generation,and rapidly reveals the essentiality of Ccdc183 in fertility,thus providing a time-saving approach for validating genetic defects(like nonsense mutations)for human infertility.

关键词： disease modeling infertility multiple morphological abnormalities of the flagella(MMAF) induction of STOP-codons(iSTOP CRISPR-STOP) sperm motility

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Coiled-coil domain-containing 38 is required for acrosome biogenesis and fibrous sheath assembly in mice

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Journal of Genetics and Genomics 2024年第4期51卷 407-418页

作者： Yaling Wang Xueying Huang Guoying Sun Jingwen Chen Bangguo Wu Jiahui Luo Shuyan Tang Peng Dai Feng Zhang jinsong li lingbo Wang Shanghai Key Laboratory of Metabolic Remodeling and Health Institute of Metabolism and Integrative BiologyFudan UniversityShanghai 200438China Institute of Reproduction and Development Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China Shanghai Key Laboratory of Maternal and Fetal Medicine Clinical and Translational Research Center of Shanghai First Maternity and Infant HospitalFrontier Science Center for Stem Cell ResearchSchool of Life Sciences and TechnologyTongji UniversityShanghai 200092China State Key Laboratory of Cell Biology CAS Center for Excellence in Molecular Cell ScienceShanghai Institute of Biochemistry and Cell BiologyChinese Academy of SciencesShanghai 200031China

During spermiogenesis,haploid spermatids undergo dramatic morphological changes to form slender sperm flagella and cap-like acrosomes,which are required for successful *** deformities in flagella cause a male infertility syndrome,multiple morphological abnormalities of the flagella(MMAF),while acrosomal hypoplasia in some cases leads to sub-optimal embryonic developmental ***,evidence regarding the occurrence of acrosomal hypoplasia in MMAF is ***,we report the generation of base-edited mice knocked out for coiled-coil domain-containing 38(Ccdc38)via inducing a nonsense mutation and find that the males are *** Ccdc38-KO sperm display acrosomal hypoplasia and typical MMAF *** find that the acrosomal membrane is loosely anchored to the nucleus and fibrous sheaths are disorganized in Ccdc38-KO *** analyses reveal that Ccdc38 knockout causes a decreased level of TEKT3,a protein associated with acrosome biogenesis,in testes and an aberrant distribution of TEKT3 in *** finally show that intracytoplasmic sperm injection overcomes Ccdc38-related *** study thus reveals a previously unknown role for CCDC38 in acrosome biogenesis and provides additional evidence for the occurrence of acrosomal hypoplasia in MMAF.

关键词： Acrosomal hypoplasia Multiple morphological abnormalities of the flagella(MMAF) Infertility Sperm motility Asthenoteratozoospermia Disease modeling

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A CRISPR/RfxC as13d-mediated strategy for efficient RNA knockdown in mouse embryonic development

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Science China life Sciences 2024年

作者： lin Zhang Shi-Meng Cao Hao Wu Meng Yan jinsong li ling-ling Chen State Key Laboratory of Cell Biology Shanghai Key Laboratory of Molecular Andrology Shanghai Institute of Biochemistry and Cell Biology Center for Excellence in Molecular Cell Science Chinese Academy of Sciences University of Chinese Academy of Sciences Key Laboratory of RNA Innovation Science and Engineering CAS Center for Excellence in Molecular Cell Science Shanghai Institute of Biochemistry and Cell Biology University of Chinese Academy of Sciences Chinese Academy of Sciences Key Laboratory of Systems Health Science of Zhejiang Province School of Life Science Hangzhou Institute for Advanced Study University of Chinese Academy of Sciences School of life Science and Technology ShanghaiTech University New Cornerstone Science Laboratory Shenzhen

The growing variety of RNA classes, such as mRNAs, lncRNAs, and circRNAs, plays pivotal roles in both developmental processes and various pathophysiological conditions. Nonetheless, our comprehension of RNA functions in live organisms remains limited due to the absence of durable and effective strategies for directly influencing RNA levels. In this study, we combined the CRISPR-RfxCas13d system with spermlike stem cell-mediated semi-cloning techniques, which enabled the suppressed expression of different RNA species. This approach was employed to interfere with the expression of three types of RNA molecules: Sfmbt2 mRNA, Fendrr lncRNA, and circMan1a2(2,3,4,5,6). The results confirmed the critical roles of these RNAs in embryonic development, as their loss led to observable phenotypes, including embryonic lethality, delayed embryonic development, and embryo resorption. In summary, our methodology offers a potent toolkit for silencing specific RNA targets in living organisms without introducing genetic alterations.

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Biallelic variants in RBM42 cause a multisystem disorder with neurological,facial,cardiac,and musculoskeletal involvement

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Protein & Cell 2024年第1期15卷 52-68页

作者： Yiyao Chen Bingxin Yang Xiaoyu Merlin Zhang Songchang Chen Minhui Wang liya Hu Nina Pan Shuyuan li Weihui Shi Zhenhua Yang li Wang Yajing Tan Jian Wang Yanlin Wang Qinghe Xing Zhonghua Ma jinsong li He-Feng Huang Jinglan Zhang Chenming Xu Institutes of Biomedical Sciences Fudan UniversityShanghai 200032China bstetrics and Gynecology Hospital Institute of Reproduction and DevelopmentFudan UniversityShanghai 200011China Intermational Peace Matermity and Child Health Hospital School of MedicineShanghai Jiao Tong UniversityShanghai 200030China Shanghai Key Laboratory of Embryo Original Diseases Shanghai 200030China State Key Laboratory of Cell Biology Shanghai Key Laboratory of Molecular AndrologyShanghai Institute of Biochemistry and Cell BiologyCAS Center for Excellence in Molecular Cell ScienceUniversity of Chinese Academy of SciencesChinese Academy of SciencesShanghai 200031China State Key Laboratory of Rice Biology the Key Laboratory of Molecular Biology of Crop Pathogens and InsectsInstitute of BiotechnologyZhejiang UniversityHangzhou 310058China Verna and Marrs McLean Department of Biochemistry and Molecular Biology Baylor College of MedicineHoustonTX 77030USA Children's hospital of Fudan University Shanghai 201102China School of life Science and Technology Shanghai Tech UniversityShanghai 201210China School of life Science Hangzhou Institute for Advanced StudyUniversity of Chinese Academy of SciencesHangzhou 310024ZhejiangChina Research Units of Embryo Original Diseases Chinese Academy of Medical Sciences(No.2019RU056)Shanghai 200011China

Here,we report a previously unrecognized syndromic neurodevelopmental disorder associated with biallelic loss-of-function variants in the RBM42 *** patient is a 2-year-old female with severe central nervous system(CNs)abnormalities,hypotonia,hearing loss,congenital heart defects,and dysmorphic facial *** whole-exome sequencing(WEs)reveals that the patient has two compound heterozygous variants,c.304C>T(p.R102*)and c.1312G>A(p.A438T),in the RBM42 gene which encodes an integral component of splicing complex in the RNA-binding motif protein *** p.A438T variant is in the RRM domain which impairs RBM42 pro-tein stability in ***,p.A438T disrupts the interaction of RBM42 with hnRNP K,which is the causa-tive gene for Au-Kline syndrome with overlapping disease characteristics seen in the index *** human R102*or A438T mutant protein failed to fully rescue the growth defects of RBM42 ortholog knockout△FgRbp1 in Fusarium while it was rescued by the wild-type(WT)human RBM42.A mouse model carying Rbm42 compound heterozygous variants,c.280C>T(p.Q94*)and c.1306_1308delinsACA(p.A436T),demonstrated gross fetal develop-mental defects and most of the double mutant animals died by ***-seq data confirmed that Rbm42 was involved in neurological and myocardial functions with an essential role in alternative splicing(As).Overall,we present clinical,genetic,and functional data to demonstrate that defects in RBM42 constitute the underlying etiology of a new neurodevelopmental disease which links the dysregulation of global AS to abnormal embryonic development.

关键词： RBM42 gene RNA-binding protein neurodevelopmental disorder Au-Kline syndrome alternative splicing

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Prognositic value of anoikis and tumor immune microenvironment-related gene in the treatment of osteosarcoma

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中南大学学报（医学版） 2024年第5期49卷 758-774页

作者： WANG Dong DENG Qing PENG Yi TONG Zhaochen li Zixin HUANG liping ZENG Jin li jinsong MIAO Jinglei CHEN Shijie Department of Spine Surgery Third Xiangya HospitalCentral South UniversityChangsha 410013 Shanghai Key Laboratory of Regulatory Biology Institute of Biomedical SciencesSchool of Life SciencesEast China Normal UniversityShanghai 200241China

Objective:Osteosarcoma is a highly aggressive primary malignant bone tumor commonly seen in children and adolescents,with a poor ***-dependent cell death(anoikis)has been proven to be indispensable in tumor metastasis,regulating the migration and adhesion of tumor cells at the primary ***,as a type of programmed cell death,anoikis is rarely studied in osteosarcoma,especially in the tumor immune *** study aims to clarify prognostic value of anoikis and tumor immune microenvironment-related gene in the treatment of ***:Anoikis-related genes(ANRGs)were obtained from *** information and ANRGs expression profiles of osteosarcoma patients were sourced from the therapeutically applicable research to generate effective therapies and Gene Expression Omnibus(GEO)*** highly associated with tumor immune microenvironment were identified by the estimate package and the weighted gene coexpression network analysis(WGCNA)*** learning algorithms were performed to construct long-term survival predictive strategy,each sample was divided into high-risk and low-risk subgroups,which was further verified in the GEO ***,based on single-cell RNA-seq from the GEO database,analysis was done on the function of signature genes in the osteosarcoma tumor ***:A total of 51 hub ANRGs closely associated with the tumor microenvironment were identified,from which 3 genes(MERTK,BNIP3,S100A8)were selected to construct the prognostic *** differences in immune cell activation and immune-related signaling pathways were observed between the high-risk and low-risk groups based on tumor microenvironment analysis(all Pling ***:The prognostic model based on ANRGs and tumor microenvironment

关键词： anoikis tumor immune microenvironment bioinformatics prognosis osteosarcoma

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Space advanced technology demonstration satellite

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Science China(Technological Sciences) 2024年第1期67卷 240-258页

作者： ZHANG XiaoFeng CHEN Wen ZHU XiaoCheng MENG Na HE JunWang BI XingZi ZHANG YongHe SHI Qi li Fei liU Rui FENG ZhengGong liU liu li jinsong WU HaiChen XU DongXiao li TaiJie HUANG JiangJiang liU Shuo li TianTong YU XianSheng GAO Yang ZHOU Heng BAN HanYu ZHANG Yanli ZHANG YueTing YANG YingQuan HE Tao DUAN Xuliang CHEN Xin WANG YaMin SUN AnTai ZHANG KuoXiang SUN Ying WANG YaoBin FAN ChengCheng XIONG Shaolin li XinQiao WEN XiangYang liNG ZhiXing SUN XiaoJin ZHANG Chen BAI XianYong WANG ZhanShan DENG YuanYong TIAN Hui YANG JianFeng XUE HongBo SANG Peng liU JinGuo ZHENG HuiLong ZHU Xiang HE JianWu li Hui XU LuXiang XU ShuYan CHEN WenWu liU ZhenDong WANG Zhaoli MAO XiangLong GAO Rong li ZongXuan DING GuoPeng WANG XinYu DOU RunJiang WENG LuBin LUO Hao WANG YaPing liANG XianFeng FANG ZiRuo Innovation Academy for Microsatellites Chinese Academy of SciencesShanghai 201203China Institute of High Energy Physics Chinese Academy of SciencesBeijing 100049China National Astronomical Observatory of China Beijing 100101China Shanghai Institute of Technical Physics Chinese Academy of SciencesShanghai 200083China Institute of Precision Optical Engineering School of Physics Science and EngineeringTongji UniversityShanghai 200092China School of Earth and Space Sciences Peking UniversityBeijing 100871China Xi’an Institute of Optics and Precision Mechanics Chinese Academy of SciencesXi’an 710119China National Space Science Center Chinese Academy of SciencesBeijing 100190China Shenyang Institute of Automation Chinese Academy of SciencesShenyang 110016China Institute of Engineering Thermophysics Chinese Academy of SciencesBeijing 100190China Institute of Mechanics Chinese Academy of SciencesBeijing 100190China Shanghai Institute of Organic Chemistry Chinese Academy of SciencesShanghai 200032China Hangzhou Institute for Advanced Study University of Chinese Academy of SciencesHangzhou 310024China Nanyang Technological University Singapore 569830Singapore Dalian Institute of Chemical Physics Chinese Academy of SciencesDalian 116023China Technical Institute of Physics and Chemistry Chinese Academy of SciencesBeijing 100049China Changchun Institute of Optics Fine Mechanics and PhysicsChinese Academy of SciencesChangchun 130033China Institute of Semiconductors Chinese Academy of SciencesBeijing 100083China Institute of Automation Chinese Academy of SciencesBeijing 100190China School of Aeronautics and Astronautics Zhejiang UniversityHangzhou 310058China

The Space Advanced Technology demonstration satellite(SATech-01),a mission for low-cost space science and new technology experiments,organized by Chinese Academy of Sciences(CAS),was successfully launched into a Sun-synchronous orbit at an altitude of~500 km on July 27,2022,from the Jiuquan Satellite Launch *** as an experimental platform for space science exploration and the demonstration of advanced common technologies in orbit,SATech-01 is equipped with 16 experimental payloads,including the solar upper transition region imager(SUTRI),the lobster eye imager for astronomy(LEIA),the high energy burst searcher(HEBS),and a High Precision Magnetic Field Measurement System based on a CPT Magnetometer(CPT).It also incorporates an imager with freeform optics,an integrated thermal imaging sensor,and a multi-functional integrated imager,*** paper provides an overview of SATech-01,including a technical description of the satellite and its scientific payloads,along with their on-orbit performance.

关键词： SATech-01 spacecraft design scientific instruments on-orbit performance

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乾嘉时期游幕诗人史善长论

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苏州科技大学学报（社会科学版） 2024年第1期41卷 64-71页

作者：李金松河南大学文学院河南开封475001

江浙士人史善长是乾嘉时期著名的游幕诗人,曾在西北幕游数年。其间,他用诗歌书写边塞的气候、风物、历史遗迹,以及自己所经历的西北回民的暴动。后来,他又入毕沅幕府,耳闻目睹发生于贵州、湖南的苗民起义与湖北的白莲教起义,并用诗歌进... 详细信息

江浙士人史善长是乾嘉时期著名的游幕诗人,曾在西北幕游数年。其间,他用诗歌书写边塞的气候、风物、历史遗迹,以及自己所经历的西北回民的暴动。后来,他又入毕沅幕府,耳闻目睹发生于贵州、湖南的苗民起义与湖北的白莲教起义,并用诗歌进行书写。史善长的这些书写,忠实记录了重要的历史事件,提供了关于这一时期重要的历史资料,不但具有文学价值,而且具有极高的史学价值,使其诗歌创作具有诗史意义。而他的诗歌风格随着幕游地的变迁也相应地发生改变,由幕游之前的“豪宕感激”变为幕游之后的“慷慨激昂”。史善长的诗歌书写在清代游幕诗人中具有典型意义。

关键词：史善长幕游边塞游幕诗人

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变尺度特征融合与交叉训练的医学报告生成方法

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计算机辅助设计与图形学学报 2024年第5期36卷 795-804页

作者：韩琪张淑军谭立玮李劲松青岛科技大学信息科学技术学院青岛266100

在对医学影像自动生成文本报告的过程中,针对病灶尺寸小、形状不规则、训练数据量少等因素易导致影像报告出现误诊、漏诊的问题,提出变尺度特征融合与交叉训练的医学报告生成方法.首先将条件全局池化后的粗粒度特征与随机丢弃后的细粒... 详细信息

在对医学影像自动生成文本报告的过程中,针对病灶尺寸小、形状不规则、训练数据量少等因素易导致影像报告出现误诊、漏诊的问题,提出变尺度特征融合与交叉训练的医学报告生成方法.首先将条件全局池化后的粗粒度特征与随机丢弃后的细粒度特征相融合,增强模型对不同尺度病灶的感知能力;然后通过整体数据和局部细节双路交叉训练的策略间接丰富数据集,增强模型的鲁棒性,并在双路中分别使用通道分离思想进一步挖掘影像的通道信息;最后通过多头注意力编解码网络,得到准确的医学报告.在IU-X-Ray和MIMIC-CXR数据集上与其他多种方法进行实验的结果表明,METEOR与BLEU-2分数分别提升5.70%和3.13%,所提方法可以有效地提升生成报告的可读性与准确性.

关键词：影像报告生成特征融合交叉训练条件池化

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精细化管理对预防接种不良反应的作用探讨

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智慧健康 2024年第16期10卷 134-136页

作者：李劲松佛山市顺德区伦教社区卫生服务中心广东佛山528300

目的探讨在预防接种过程中精细化管理对不良反应的影响。方法选择本中心2022年1月—2023年5月服务的108例接种儿童为研究对象,随机将其分为两组,每组54例。对照组行常规管理,研究组行精细化管理。探讨两种不同护理方法在预防接种过程中... 详细信息

目的探讨在预防接种过程中精细化管理对不良反应的影响。方法选择本中心2022年1月—2023年5月服务的108例接种儿童为研究对象,随机将其分为两组,每组54例。对照组行常规管理,研究组行精细化管理。探讨两种不同护理方法在预防接种过程中的接种情况、不良反应率以及满意度。结果研究组的及时接种率比对照组更高,纠纷发生率比对照组更低,组间差异显著,具有统计学意义(P<0.05);研究组的不良反应发生率低于对照组,组间差异显著,具有统计学意义(P<0.05);研究组的儿童家长对护理服务满意度高于对照组,组间差异显著,具有统计学意义(P<0.05)。结论在预防接种过程中,在儿童预防接种过程中开展精细化管理,可以有效地提高及时接种率、护理满意度,降低纠纷发生率、不良反应发生率,是一项值得临床关注的工作。

关键词：精细化管理疫苗接种不良反应

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人诺如病毒感染引起的急性胃肠炎流行特征及其影响因素的研究进展

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病毒学报 2024年第1期40卷 140-150页

作者：谢迪何莲花刘晖李金松成都康华生物制品股份有限公司成都610100 传染病溯源预警与智能决策全国重点实验室国家卫生健康委员会医学病毒与病毒病重点实验室中国疾病预防控制中心病毒病预防控制所北京102206 甘肃中医药大学公共卫生学院兰州730000

人诺如病毒(Human norovirus, HuNoV)属于杯状病毒科诺如病毒属,为无包膜单股正链RNA病毒,易发生重组,变异快,传染性强,致病剂量低;人群普遍易感,是全球急性胃肠炎(Acute gastroenteritis, AGE)的主要病因之一。HuNoV感染后,≤5岁儿童和... 详细信息

人诺如病毒(Human norovirus, HuNoV)属于杯状病毒科诺如病毒属,为无包膜单股正链RNA病毒,易发生重组,变异快,传染性强,致病剂量低;人群普遍易感,是全球急性胃肠炎(Acute gastroenteritis, AGE)的主要病因之一。HuNoV感染后,≤5岁儿童和>65岁老年人群中少数人会因大量失水导致电解质紊乱等重症,甚至死亡。HuNoV还易在封闭和/或半封闭人群拥挤的场所引起AGE暴发;新的变异株或/和型别出现,会导致暴发事件大幅升高,甚至出现大流行。因此,HuNoV导致了沉重的疾病负担,是全球性的公共卫生问题。本文综述了全球HuNoV AGE的流行病学研究进展,还从宿主、病毒和环境等多个方面综述了当前影响HuNoV AGE流行特征的主要影响因素,为HuNoV AGE的防控提供支撑。

关键词：人诺如病毒急性胃肠炎流行特征影响因素

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