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Multiplex CRISPR/Cas9-mediated knockout of soybean LNK2 advances flowering time

2016年全国植物生物学大会

作者： baohui liu Key Laboratory of Soybean Molecular Design Breeding Northeast Institute of Geography and AgroecologyChinese Academy of Sciences

Photoperiodism is a rhythmic change of sensitivity to light,which helps plants to adjust flowering time accor to seasonal changes in day length and to adapt to growing conditions at various latitudes. To reveal the molecular basis of photoperiod is min soybean(Glycine max),a facultative short-day plant,we analyzed the transcriptional profiles of the maturity gene E1 family and two FLOWERING LOCUS T(FT) orthologs(FT2a and FT5a). E1,a repressor for FT2 a and FT5 a,and its two homologs,E1-like-a(E1La) and E1 Lb,exhibited two peaks of expression in long days. Using two different approaches(experiments with transition between light and dark phases and night-break experiments),we revealed that the E1 family genes were expressed only during light periods and that their induction after dawn in long days required a period of light before dusk the previous day. In the cultivar Toyomusume,which lacks the E1 gene,virus-induced silencing of E1 La and E1 Lb up-regulated the expression of FT2 a and FT5 a and led to early flowering. Therefore,E1,E1 La,and E1 Lb function similarly in flowering. Regulation of E1 and E1 L expression by light was under the control of E3 and E4,which encode phytochrome A proteins. Our data suggest that phytochrome A-mediated transcriptional induction of E1 and its homologs by light plays a critical role in photoperiodic induction of flowering in soybean.

关键词： photoperiodism soybean flowering E1 gene

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The Crop Journal 2021年第4期9卷 767-776页

作者： Zhaobo Li Qun Cheng Zhuoran Gan Zhihong Hou Yuhang Zhang Yongli Li Haiyang Li Haiyang Nan Cen Yang Linnan Chen Sijia Lu Wenqian Shi Liyu Chen Yanping Wang Chao Fang Liping Kong Tong Su Shichen Li Kun Kou Lingshuang Wang Fanjiang Kong baohui liu Lidong Dong The Innovative Academy of Seed Design Key Laboratory of Soybean Molecular Design BreedingNortheast Institute of Geography and AgroecologyChinese Academy of SciencesHarbin 130102HeilongjiangChina University of Chinese Academy of Sciences Beijing 100081China Innovation Center of Molecular Genetics and Evolution School of Life SciencesGuangzhou UniversityGuangzhou 510006GuangdongChina Agronomy College of Heilongjiang Bayi Agricultural University Daqing 163319HeilongjiangChina National Center for Soybean Improvement National Key Laboratory of Crop Genetics and Germplasm EnhancementJiangsu Collaborative Innovation Center for Modern Crop ProductionNanjing Agricultural UniversityNanjing 210095JiangsuChina Mudanjiang Branch Heilongjiang Academy of Agricultural SciencesMudanjiang 157000HeilongjiangChina

Flowering time is an important agronomic trait for soybean yield and adaptation. However, the genetic basis of soybean adaptation to diverse latitudes is still not clear. Four NIGHT LIGHT-INDUCIBLE AND CLOCK-REGULATED 2(LNK2) homeologs of Arabidopsis thaliana LNK2 were identified in soybean. Three single-guide RNAs were designed for editing the four LNK2 genes. A transgene-free homozygous quadruple mutant of the LNK2 genes was developed using the CRISPR(clustered regularly interspaced short palindromic repeats)/Cas9(CRISPR-associated protein 9). Under long-day(LD) conditions, the quadruple mutant flowered significantly earlier than the wild-type(WT). Quantitative real-time PCR(q RT-PCR)revealed that transcript levels of LNK2 were significantly lower in the quadruple mutant than in the WT under LD conditions. LNK2 promoted the expression of the legume-specific E1 gene and repressed the expression of FT2 a. Genetic markers were developed to identify LNK2 mutants for soybean breeding.These results indicate that CRISPR/Cas9-mediated targeted mutagenesis of four LNK2 genes shortens flowering time in soybean. Our findings identify novel components in flowering-time control in soybean and may be beneficial for further soybean breeding in high-latitude environments.

关键词： Soybean LNK2 CRISPR/Cas9 Genome editing Flowering time

The incidence rate and histological characteristics of intimal hyperplasia in elastase-induced experimental abdominal aortic aneurysms in mice

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Animal Models and Experimental Medicine 2024年第3期7卷 388-395页

作者： Meng Li Panpan Wei Kexin Li Haole liu Naqash Alam Haiwen Hou Jie Deng baohui Xu Enqi liu Sihai Zhao Yankui Li Laboratory Animal Center Xi'an Jiaotong UniversityXi'anChina Department of Vascular Surgery The Second Hospital of Tianjin Medical UniversityTianjinChina Department of Cardiology The Second Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina Department of Vascular Surgery Stanford University School of MedicineStanfordCaliforniaUSA

Intimal hyperplasia(IH)is a negative vascular remodeling after arterial injury.IH occasionally occurs in elastase-induced abdominal aortic aneurysm(AAA)mouse models.This study aims to clarify the incidence and histological characteristics of IH in aneurysmal mice.A retrospective study was conducted by including 42 male elastaseinduced mouse AAA models.The IH incidence,aortic diameters with or without IH,and hyperplasia lesional features of mice were analyzed.Among 42 elastase-induced AAA mouse models,10 mice developed mild IH(24%)and severe IH was found in only 2 mice(5%).The outer diameters of the AAA segments in mice with and without IH did not show significant difference.Both mild and severe IH lesions show strong smooth muscle cell positive staining,but endothelial cells were occasionally observed in severe IH lesions.There was obvious macrophage infiltration in the IH lesions of the AAA mouse models,especially in mice with severe IH.However,only a lower numbers of T cells and B cells were found in the IH lesion.Local cell-secreted matrix metalloproteinases(MMP)2 was highly expressed in all IH lesions,but MMP9 was only overexpressed in severe lesions.In conclusion,this study is the first to demonstrate the occurrence of aneurysmal IH and its histological characteristics in an elastaseinduced mouse AAA model.This will help researchers better understand this model,and optimize it for use in AAA-related research.

关键词： abdominal aortic aneurysms animal model histology intimal hyperplasia

Molecular mechanism underlying photoperiod regulated flowering in soybean

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Molecular mechanism underlying photoperiod regulated floweri...

Oncogenic β-catenin-driven liver cancer is susceptible to methotrexate-mediated disruption of nucleotide synthesis

第十四届全国植物基因组学大会

作者： Fanjiang Kong Abe Jun baohui liu Key Laboratory of Soybean Molecular Design Breeding Northeast Institute of Geography and AgroecologyChinese Academy of Sciences Research Faculty of Agriculture Hokkaido University

Flowering is one of the most important processes involved in crop adaptation and productivity.A number of major genes and quantitative trait loci(QTLs) for flowering have been reported in soybean(Glycine max).These ge... 详细信息

关键词： soybean QTL flowering maturity yield adaptation

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Chinese Medical Journal 2024年第2期137卷 181-189页

作者： Fangming liu Yuting Wu baohui Zhang Shuhui Yang Kezhuo Shang Jie Li Pengju Zhang Weiwei Deng Linlin Chen Liang Zheng Xiaochen Gai Hongbing Zhang Department of Physiology State Key Laboratory of Common Mechanism Research for Major DiseasesHaihe Laboratory of Cell EcosystemInstitute of Basic Medical Sciences and School of Basic MedicineChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100005China Department of Physiology Institute of Basic Medical Sciences and School of Basic MedicineGraduate School of Peking Union Medical CollegeChinese Academy of Medical SciencesBeijing100005China Department of Physiology School of Life ScienceChina Medical UniversityShenyangLiaoning 110122China School of Life Sciences Westlake UniversityHangzhouZhejiang 310024China Key Laboratory of Pediatric Hematology and Oncology Ministry of HealthInstitute of Pediatric Translational MedicineShanghai Children’s Medical CenterShanghai Jiao Tong School of MedicineShanghai 200127China

Background:Liver cancer is largely resistant to chemotherapy.This study aimed to identify the effective chemotherapeutics forβ-catenin-activated liver cancer which is caused by gain-of-function mutation of catenin beta 1(CTNNB1),the most frequently altered proto-oncogene in hepatic neoplasms.Methods:Constitutiveβ-catenin-activated mouse embryonic fibroblasts(MEFs)were established by deleting exon 3(β-catenin^(Δ(ex3)/+)),the most common mutation site in CTNNB1 gene.A screening of 12 widely used chemotherapy drugs was conducted for the ones that selectively inhibitedβ-catenin^(Δ(ex3)/+)but not for wild-type MEFs.Untargeted metabolomics was carried out to examine the alterations of metabolites in nucleotide synthesis.The efficacy and selectivity of methotrexate(MTX)onβ-catenin-activated human liver cancer cells were determined in vitro.Immuno-deficient nude mice subcutaneously inoculated withβ-catenin wild-type or mutant liver cancer cells and hepatitis B virus(HBV);β-catenin^(lox(ex3)/+)mice were used,respectively,to evaluate the efficacy of MTX in the treatment ofβ-catenin mutant liver cancer.Results:MTX was identified and validated as a preferential agent against the proliferation and tumor formation ofβ-catenin-activated cells.Boosted nucleotide synthesis was the major metabolic aberration inβ-catenin-active cells,and this alteration was also the target of MTX.Moreover,MTX abrogated hepatocarcinogenesis of HBV;β-catenin^(lox(ex3)/+)mice,which stimulated concurrent Ctnnb1-activated mutation and HBV infection in liver cancer.Conclusion:MTX is a promising chemotherapeutic agent forβ-catenin hyperactive liver cancer.Since repurposing MTX has the advantages of lower risk,shorter timelines,and less investment in drug discovery and development,a clinical trial is warranted to test its efficacy in the treatment ofβ-catenin mutant liver cancer.

关键词： Liver cancer β-catenin Methotrexate Nucleotide synthesis Chemotherapy

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基于边光滑有限元法的骨重建力电效应

医用生物力学 2022年第4期37卷 631-637页

作者：朱婷婷刘宝会王刚刘易河北工业大学机械工程学院天津300401 国家技术创新方法与实施工具工程技术研究中心天津300401 天津市北辰区中医医院急诊科天津300499

目的针对骨重建压电效应仿真分析时传统有限元法(finite element method,FEM)数值解精度差的问题,提出了一种基于边的光滑有限元法(edged-based smoothed FEM,ES-FEM)模型。方法基于三角形背景网格构建光滑域,依托梯度光滑技术获得光滑... 详细信息

目的针对骨重建压电效应仿真分析时传统有限元法(finite element method,FEM)数值解精度差的问题,提出了一种基于边的光滑有限元法(edged-based smoothed FEM,ES-FEM)模型。方法基于三角形背景网格构建光滑域,依托梯度光滑技术获得光滑应变梯度与光滑电场梯度,在光滑伽辽金弱形式的框架下构造系统离散方程。结果采用上述模型能反映压电效应下骨重建过程中骨密度变化和电势分布,相比于FEM,ES-FEM能够在一定程度上提高骨重建仿真结果的精度。结论提出的边光滑有限元模型能够更准确地模拟出骨重建过程,该方法对骨重建压电效应问题的准确预测为骨类疾病临床研究提供有效的理论依据。

关键词：骨重建边光滑有限元法梯度光滑技术压电效应数值算法

Novel RNAIII-Inhibiting Peptide Derivatives Increase Survival in Methicillin-Resistant Staphylococcus aureus Sepsis by Inhibiting NLRP3 Inflammasome Activation

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Novel RNAIII-Inhibiting Peptide Derivatives Increase Surviva...

雄安高铁站站房及雨棚金属屋面设计与关键技术应用

2013年中国药学大会暨第十三届中国药师周

作者： Zheng Hou Ying Zhou Zhi Li Qian Yu Mingkai Li Xiaoyan Xue Fei Da Di Qu Xin Shi baohui liu Xiaoxing Luo Department of Pharmacology School of Pharmacy Fourth Military Medical University

Background. Previous researches have shown RNAIII-inhibiting peptide (RIP) could effectively inhibit the quorum sensing (QS) system in staphylococci. However, it remains ill-understood why RIP could markedly increase the survival rate in vivo while not influencing the number of bacteria in vitro. Methods. Both inflammasome formation and the release of mature IL-1β were assessed after treatment with RIP derivatives in a murine sepsis model. Secretion of hemolysins, survival rates, bacterial titers and histological changes were determined. Results. The secretion of hemolysins and the mRNA expression of RNAIII, NLRP3 and ASC were markedly suppressed by RIPs in vivo. Caspase-1 and IL-1β protein levels were reduced. RIP1183 produced the greatest improvement in mice survival and inhibition of pathological damage. The systemic application of RIP1183 was even superior to that of vancomycin in MRSA-infected mice. No antibacterial activities were displayed by any RIP in vitro. Conclusions. We firstly demonstrated interfering with cell-cell communication by using a quorum-sensing inhibitor-RIP1183 can markedly inhibit the activation of the inflammasome signal pathway and refrain from excessive inflammatory response. Anti-virulence strategies targeting bacterial QS system may be a novel development direction of antimicrobial agents, which avoided the excessive bacterial survival pressure and the induction of drug resistance.

关键词： Methicillin-resistant Staphylococcus aureus （MRSA） RNAIII-inhibiting peptide （RIP） NLRP3 inflammasome Quorum sensing （QS）

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中国建筑防水 2022年第4期 26-30页

作者：刘宝辉森特士兴集团股份有限公司北京100176

雄安高铁站金属屋面设计包含了各分系统的设计,各分系统之间相互关联且有效连接,组成整体功能性的屋面系统。本文介绍了该金属屋面工程玻璃采光顶、直立锁边+分布式光伏、阳光板+分布式光伏、檐口装饰板等4大系统构造设计及特点,以及为... 详细信息

雄安高铁站金属屋面设计包含了各分系统的设计,各分系统之间相互关联且有效连接,组成整体功能性的屋面系统。本文介绍了该金属屋面工程玻璃采光顶、直立锁边+分布式光伏、阳光板+分布式光伏、檐口装饰板等4大系统构造设计及特点,以及为满足设计要求和解决工程难点而应用的BIM协同技术、金属屋面加设分布式光伏、高飞翼阳光板成套技术等多项关键技术,为同类工程提供了借鉴。

关键词：高铁站房金属屋面系统构造设计 BIM协同技术分布式光伏高飞翼阳光板

D-二聚体对成年烧伤患者下肢深静脉血栓形成的预测价值

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中华烧伤与创面修复杂志 2022年第4期38卷 335-340页

作者：张伟刘宝辉夏成德丘宁宁娄季鹤狄海萍薛继东李刚郑州市第一人民医院介入科郑州450004 郑州市第一人民医院烧伤科郑州450004

目的探讨D-二聚体对成年烧伤患者并发下肢深静脉血栓形成(DVT)的预测价值。方法采用回顾性病例系列研究方法。收集2015年1月1日—2019年12月31日郑州市第一人民医院烧伤科收治的符合入选标准的3861例成年烧伤患者的临床资料。按住院期... 详细信息

目的探讨D-二聚体对成年烧伤患者并发下肢深静脉血栓形成(DVT)的预测价值。方法采用回顾性病例系列研究方法。收集2015年1月1日—2019年12月31日郑州市第一人民医院烧伤科收治的符合入选标准的3861例成年烧伤患者的临床资料。按住院期间是否并发下肢DVT,将患者分为DVT组(77例)和非DVT组(3784例),收集并比较2组患者的性别、年龄、烧伤总面积、D-二聚体水平,入院时有无下肢烧伤、吸入性损伤,住院期间是否并发脓毒症/感染性休克及有无股静脉留置中心静脉导管(CVC)、外科手术史、输注浓缩红细胞。对数据行独立样本t检验、Mann-Whitney U检验、χ2检验。选取组间比较差异具有统计学意义的指标进行多因素logistic回归分析,筛选3861例成年烧伤患者并发下肢DVT的独立危险因素。绘制独立危险因素预测3861例成年烧伤患者并发下肢DVT的受试者操作特征(ROC)曲线,计算曲线下面积(AUC)、最佳阈值及最佳阈值下的敏感度、特异度。采用Delong检验对AUC的质量进行比较,采用χ2检验对最佳阈值下的敏感度和特异度进行比较。结果2组患者性别及住院期间脓毒症/感染性休克发生情况、外科手术史比较,差异均无统计学意义(P>0.05);2组患者年龄、烧伤总面积、D-二聚体水平,入院时下肢烧伤情况、吸入性损伤情况,住院期间股静脉留置CVC和输注浓缩红细胞情况比较,差异均有统计学意义(t=-8.17,Z值分别为-5.04、-10.83,χ2值分别为21.83、5.37、7.75、4.52,P0.05)。结论D-二聚体水平是成年烧伤患者并发下肢DVT的独立危险因素,其AUC质量及最佳阈值下的敏感度、特异度优于年龄和烧伤总面积,对成年烧伤患者并发下肢DVT具有较好预测价值。

关键词：烧伤下肢静脉血栓形成预测 D-二聚体

早期剖宫产切口部妊娠患者侧支供血的影响因素分析

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中华医学杂志 2022年第2期102卷 130-135页

作者：张伟李刚华成飞王海波张同庆陈清亮梁昊刘宝辉郑州市第一人民医院介入科郑州450004 郑州市中心医院外周介入科郑州450007 郑州市第三人民医院介入科郑州450000 河南省直第三人民医院放射科郑州450006

目的分析早期剖宫产切口部妊娠(CSP)患者侧支供血的发生情况及影响因素。方法本研究为多中心的病例对照研究,以郑州市第一人民医院、郑州市中心医院、郑州市第三人民医院、河南省直第三人民医院2017年1月1日至2020年6月30日219例CSP住... 详细信息

目的分析早期剖宫产切口部妊娠(CSP)患者侧支供血的发生情况及影响因素。方法本研究为多中心的病例对照研究,以郑州市第一人民医院、郑州市中心医院、郑州市第三人民医院、河南省直第三人民医院2017年1月1日至2020年6月30日219例CSP住院患者为研究对象,获取其临床资料,数字减影血管造影影像分析采用双盲法,将研究对象分为有侧支供血组和无侧支供血组,计算早期CSP侧支供血的发生率,采用多因素logistic回归模型分析早期CSP侧支供血发生的影响因素,同时比较两组患者子宫动脉栓塞术(UAE)后的临床结局情况。结果219例早期CSP患者年龄为(32.4±5.0)岁,孕期为(51.0±10.6)d,早期CSP侧支供血发生率为12.3%(27例),发生于左侧者16例,右侧者6例,双侧者5例;共发现43条侧支供血血管,平均每患者1.59条,侧支供血血管来源以膀胱动脉最多见,占74.4%(32/43),其次为阴部内动脉,占18.6%(8/43)。多因素分析结果显示,孕周≥8周、妊娠囊最大径≥50 mm、妊娠囊呈富血供型是早期CSP侧支供血发生的影响因素,OR值(95%CI)分别为3.68(1.06~12.76)、7.00(1.52~32.19)、9.96(3.59~27.58),均P0.05)。结论早期CSP具有一定的侧支供血发生率,有可能对UAE的疗效及患者安全产生不利影响;孕周、妊娠囊最大径及其血管化程度在提示早期CSP侧支供血上具有一定的价值,有助于UAE过程中对妊娠囊的完全栓塞。

关键词：妊娠,异位血管造影术,数字减影剖宫产切口部妊娠血液供应