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In vivo evaluation of guide-free Cas9-induced safety risks in a pig model

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Signal Transduction and Targeted Therapy 2024年第8期9卷 3493-3506页

作者： Weikai Ge Shixue Gou Xiaozhu Zhao Qin Jin Zhenpeng Zhuang Yu Zhao Yanhui Liang Zhen ouyang Xiaoyi Liul Fangbing Chen Hui Shi Haizhao Yan Han wu Liangxue Lai Kepin Wang China-New Zealand Joint Laboratory on Biomedicine and Health CAS Key Laboratory of Regenerative BiologyGuangdong Provincial Key Laboratory of Stem Cell and Regenerative MedicineJoint School of Life SciencesGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou Medical UniversityGuangzhou 510530China Sanya institute of Swine resource Hainan Provincial Research Center of Laboratory AnimalsSanya 572000China Guangdong Provincial Key Laboratory of Large Animal models for Biomedicine Wuyi UniversityJiangmen 529020China Guangzhou National Laboratory Guangzhou 510005China SUniversity of Chinese Academy of Sciences 100049 BeijingChina Research Unit of Generation of Large Animal Disease Models Chinese Academy of Medical Sciences(2019RU015)Guangzhou 510530China

The CRISPR/Cas9 system has shown great potential for treating human genetic diseases through gene ***,there are concerns about the safety of this system,specifically related to the use of guide-free *** studies have shown that guidefree Cas9 can induce genomic instability in ***,the in vivo safety risks associated with guide-free Cas9 have not been evaluated,which is necessary for the development of gene therapy in clinical *** this study,we used doxycycline-inducible Cas9-expressing pigs to evaluate the safety risks of guide-free Cas9 in *** findings demonstrated that expression of guide-free Cas9 could induce genomic damages and transcriptome changes in *** severity of the genomic damages and transcriptome changes were correlate with the expression levels of Cas9 ***,prolonged expression of Cas9 in pigs led to abnormal phenotypes,including a significant decrease in body weight,which may be attributable to genomic damage-induced nutritional absorption and metabolic ***,we observed an increase in whole-genome and tumor driver gene mutations in pigs with long-term Cas9 expression,raising the risk of tumor *** in vivo evaluation of guide-free Cas9 in pigs highlights the necessity of considering and monitoring the detrimental effects of Cas9 alone as genome editing via the CRISPR/Cas9 system is implemented in clinical gene *** research emphasizes the importance of further study and implementation of safety measures to ensure the successful and safe application of the CRiSPR/Cas9 system in clinical practice.

关键词： CRISPR/Cas9 vivo alone

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伊曲康唑纳米乳的制备及含量测定

西北农林科技大学学报（自然科学版） 2014年第8期42卷 34-39,44页

作者：高瑞欧阳五庆西北农林科技大学动物医学院陕西杨凌712100

【目的】研制伊曲康唑纳米乳并评价其质量,考察纳米乳中伊曲康唑含量。【方法】以伊曲康唑为主要活性成分,肉桂醛和乙酸乙酯为混合油相,EL-40为表面活性剂,异丙醇为助表面活性剂,通过绘制伪三元相图确定伊曲康唑纳米乳的最优配方。利用... 详细信息

【目的】研制伊曲康唑纳米乳并评价其质量,考察纳米乳中伊曲康唑含量。【方法】以伊曲康唑为主要活性成分,肉桂醛和乙酸乙酯为混合油相,EL-40为表面活性剂,异丙醇为助表面活性剂,通过绘制伪三元相图确定伊曲康唑纳米乳的最优配方。利用透射电子显微镜(TEM)、激光粒度分析仪等评价伊曲康唑纳米乳的质量,利用紫外分光光度计考察纳米乳中伊曲康唑含量。【结果】经筛选,伊曲康唑纳米乳配方中各物质的质量分数为:伊曲康唑1.24%,肉桂醛4.58%,乙酸乙酯4.58%,EL-40 24.44%,异丙醇12.17%,蒸馏水52.99%。在TEM下,伊曲康唑纳米乳乳滴为圆球形,分布均一,平均粒径为14.48nm,多分散系数(PDI)为0.014。制备出的纳米乳澄清透明、稳定性良好。测定伊曲康唑纳米乳中伊曲康唑含量的最佳波长为261nm,检测方法的专属性良好。【结论】伊曲康唑纳米乳稳定性好且制备工艺简单,用于测定伊曲康唑纳米乳中伊曲康唑含量的紫外分光光度法专属性好、精密度高。

关键词：伊曲康唑纳米乳质量评价

Continuous age-and sex-specific reference ranges of liver enzymes in Chinese children and application in pediatric non-alcoholic fatty liver disease

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World Journal of Pediatrics 2024年第9期20卷 949-956页

作者： Zhao-Yuan wu Si-Wei Chi Liu-Jian ouyang Xiao-Qin Xu Jing-Nan Chen Bing-Han Jin Rahim Ullah Xue-Lian Zhou Ke Huang Guan-Ping Dong Zhe-Ming Li Ying Shen Jie Shao Yan Ni Jun-Fen Fu Qiang Shu Wei wu Department of Endocrinology Children’s HospitalZhejiang University School of MedicineNational Clinical Research Center for Child Health3333 Binsheng RoadHangzhou310051China Department of Data and Information Children’s HospitalZhejiang University School of MedicineHangzhouChina Department of Child Health Care Children’s HospitalZhejiang University School of MedicineNational Clinical Research Center for Child HealthHangzhouChina Department of Thoracic&Cardiovascular Surgery Children’s HospitalZhejiang University School of MedicineNational Clinical Research Center for Child HealthNational Children’s Regional Medical Center3333 Binsheng RoadHangzhou310051China

Background Alanine aminotransferase(ALT)is widely used to screen patients with hepatic ***,the current reference ranges(<50 U/L)were developed by laboratories and have not been validated in populations with a large number of healthy *** This study collected venous blood and anthropometric data from a total of 13,287 healthy children aged 3 months to 18 years who underwent routine physical examinations in the Department of Pediatric *** applied the least mean square algorithm to establish age-and sex-related reference percentiles of serum levels of *** validation,we recruited 4276 children and adolescents with obesity/overweight who underwent evaluation and metabolic tests in the *** receiver operating characteristic curves,we determined age-and sex-specific upper limit percentiles of liver enzymes for fatty liver *** This study revealed a significant correlation between serum transaminase levels and age and sex(P<0.01).These transaminase levels exhibited age-and sex-specific *** individuals in the non-alcoholic fatty liver disease(NAFLD)cohort,elevated ALT levels displayed a positive association with clinical markers of disease severity,including homeostatic model assessment of insulin resistance,waist-hip ratio,and serum uric acid levels(P<0.01).According to the receiver operating characteristic curves,ALT levels at the 92.58th percentile for boys and the 92.07th percentile for girls yielded the highest accuracy and *** This study provides age-and sex-specific reference ranges for ALT,aspartate aminotransferase,andγ-glutamyltransferase in Chinese children and adolescents,making it the largest population study to ***,the study establishes a precise upper limit for ALT levels,facilitating their use in NAFLD screening.

关键词： Alanine aminotransferase Childhood Fatty liver diseases

基于配电网节点边际容量成本的分布式电源规划

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电力系统自动化 2009年第18期33卷 28-32页

作者：欧阳武程浩忠张秀彬上海交通大学电子信息与电气工程学院上海市200240

分布式电源(DG)接入配电网可能导致现有供电设备得不到充分利用,产生沉没成本。引入配电网节点边际容量成本(LMCC)以反映各节点容量的紧张程度,同时给出DG单位容量成本的计算方法。在此基础上,以DG单位容量成本小于接入点的边际容量成... 详细信息

分布式电源(DG)接入配电网可能导致现有供电设备得不到充分利用,产生沉没成本。引入配电网节点边际容量成本(LMCC)以反映各节点容量的紧张程度,同时给出DG单位容量成本的计算方法。在此基础上,以DG单位容量成本小于接入点的边际容量成本作为DG接入的判据,提出一种规划DG位置和容量的新方法。算例结果验证了方法的可行性。

关键词：配电网分布式电源电源规划节点边际容量成本供电容量

盐酸黄连素纳米微乳的制备及其体外抗菌活性研究

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西北农林科技大学学报（自然科学版） 2007年第1期35卷 54-57,62页

作者：孙红武欧阳五庆西北农林科技大学动物科技学院陕西杨凌712100

用高效液相色谱仪测定了盐酸黄连素在4种油(IPM、大豆油、花生油、色拉油)和3种表面活性剂(EL 40、大豆卵磷脂和Tw een-80)中的溶解度;根据纳米微乳的评价标准,利用序贯设计思想,采用滴定法绘制了伪三元相图,确定了纳米微乳的最佳组成,... 详细信息

用高效液相色谱仪测定了盐酸黄连素在4种油(IPM、大豆油、花生油、色拉油)和3种表面活性剂(EL 40、大豆卵磷脂和Tw een-80)中的溶解度;根据纳米微乳的评价标准,利用序贯设计思想,采用滴定法绘制了伪三元相图,确定了纳米微乳的最佳组成,研制了盐酸黄连素纳米微乳,并对其基本理化特性(形态、粒度、颜色、澄清度、稳定性和自乳化速度等)及体外抗菌活性进行了研究。结果显示,盐酸黄连素纳米微乳三相最佳组成是EL 40-甘油-IPM;制得的纳米微乳为澄清透明的球状液滴,其平均粒径为56.8 nm;高速离心、高温、强光下无分层,也无絮凝或药物析出;与传统片剂、胶囊和水溶液对大肠杆菌、沙门氏菌、金黄色葡萄球菌和无乳链球菌的抗菌活性相比,纳米微乳的抗菌活性更高。以上研究表明制备的盐酸黄连素纳米微乳是一种质量稳定、高效的抗菌剂。

关键词：黄连素纳米微乳稳定性抗菌活性

轮缘推进器水润滑橡胶弹支可倾瓦推力轴承均载特性

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交通运输工程学报 2021年第2期21卷 138-149页

作者：宁昶雄严新平欧阳武武汉理工大学能源与动力工程学院湖北武汉430063 武汉理工大学国家水运安全工程技术研究中心湖北武汉430063 武汉理工大学船舶动力工程技术交通行业重点实验室湖北武汉430063

为了探讨船舶轮缘推进器(RDT)橡胶垫支撑水润滑推力轴承的均载特性,提出了推力轴承均载特性参数测试方法;在多功能立式水润滑试验台上,以用于RDT的内径124 mm、外径196 mm水润滑橡胶垫支撑推力轴承为试验对象,在盘面上选取轴承平均半径... 详细信息

为了探讨船舶轮缘推进器(RDT)橡胶垫支撑水润滑推力轴承的均载特性,提出了推力轴承均载特性参数测试方法;在多功能立式水润滑试验台上,以用于RDT的内径124 mm、外径196 mm水润滑橡胶垫支撑推力轴承为试验对象,在盘面上选取轴承平均半径的截面,对称布置1个微型压力传感器和1个微型温度传感器,随着轴一起旋转,采用无线遥测技术分别获取全瓦水膜压力分布和推力盘温度;通过预设瓦块高度差和推力盘静态倾斜量模拟偏载的情况,研究了载荷和转速变化对试验轴承水膜压力分布、摩擦因数和推力盘温度的影响规律。研究结果表明:弹支的均载效果会随着工况的变化而变化,当转速不变时,载荷增大会增加各瓦橡胶垫的变形,从而增强均载效果;而推力盘倾斜程度会随着转速增加而增强,从而加剧了瓦块载荷的不均性;开展RDT橡胶弹支可倾瓦结构均载设计时,除了考虑推力盘和瓦块不平的制造和安装因素,还需考虑轴承的转速和载荷;从轴承各瓦压力分布随工况变化的关系看,在转速为100 r·min-1、载荷为0.35 MPa时,轴承接触承载比例升高,因此,水膜压力测试为判别轴承润滑状态提供了一条新途径。

关键词：轮缘推进器水润滑弹支可倾瓦推力轴承无线遥测压力分布润滑状态均载特性

High-yield and rapid isolation of extracellular vesicles by flocculation via orbital acoustic trapping:FLOAT

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Microsystems & Nanoengineering 2024年第1期10卷 313-323页

作者： Joseph Rufo Peiran Zhang Zeyu Wang Yuyang Gu Kaichun Yang Joseph Rich Chuyi Chen Ruoyu Zhong Ke Jin Ye He Jianping Xia Ke Li Jiarong wu Yingshi ouyang Yoel Sadovsky Luke P.Lee Tony Jun Huang Department of Mechanical Engineering and Materials Science Duke UniversityDurhamNCUSA Department of Biomedical Engineering Duke UniversityDurhamNCUSA Department of Obstetrics Gynecologyand Reproductive SciencesMagee-Womens Research InstituteUniversity of PittsburghPittsburghPAUSA Department of Microbiology and Molecular Genetics School of MedicineUniversity of PittsburghPittsburghPAUSA Renal Division and Division of Engineering in Medicine Department of MedicineBrigham and Women’s HospitalHarvard Medical SchoolBostonMAUSA Department of Bioengineering Department of Electrical Engineering and Computer ScienceUniversity of CaliforniaBerkeleyBerkeleyCAUSA Department of Biophysics Institute of Quantum BiophysicsSungkyunkwan UniversitySuwonKorea

Extracellular vesicles(EVs)have been identified as promising biomarkers for the noninvasive diagnosis of various ***,challenges in separating EVs from soluble proteins have resulted in variable EV recovery rates and low ***,we report a high-yield(>90%)and rapid(<10 min)EV isolation method called FLocculation via Orbital Acoustic Trapping(FLOAT).The FLOAT approach utilizes an acoustofluidic droplet centrifuge to rotate and controllably heat liquid *** adding a thermoresponsive polymer flocculant,nanoparticles as small as 20 nm can be rapidly and selectively concentrated at the center of the *** demonstrate the ability of FLOAT to separate urinary EVs from the highly abundant Tamm-Horsfall protein,addressing a significant obstacle in the development of EV-based liquid *** to its high-yield nature,FLOAT reduces biofluid starting volume requirements by a factor of 100(from 20 mL to 200µL),demonstrating its promising potential in point-of-care diagnostics.

关键词： trapping utilize separating

船舶新型水润滑磁液双浮尾轴承承载能力仿真研究

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船舶力学 2022年第9期26卷 1326-1334页

作者：李哲欧阳武汪盛通何涛王斌武汉理工大学交通与物流工程学院武汉430063 船舶动力工程技术交通行业重点实验室武汉430063 国家水运安全工程技术研究中心可靠性工程研究所武汉430063 武汉第二船舶设计研究所武汉430205 武汉理工大学船海与能源动力工程学院武汉430063

为提高船舶水润滑尾轴承的承载能力,提出新型水润滑磁液双浮尾轴承方案。设计水润滑磁液双浮尾轴承结构,建立双浮轴承磁-流-固多场耦合动力学模型。提出六种磁路布置方案,采用有限元法进行计算。对比各方案磁力,优选磁斥力最大方案。在... 详细信息

为提高船舶水润滑尾轴承的承载能力,提出新型水润滑磁液双浮尾轴承方案。设计水润滑磁液双浮尾轴承结构,建立双浮轴承磁-流-固多场耦合动力学模型。提出六种磁路布置方案,采用有限元法进行计算。对比各方案磁力,优选磁斥力最大方案。在此基础上,采用有限差分法计算液膜力,与磁斥力叠加得到轴承总承载力,并分析其随轴承间隙的变化规律。研究结果表明:与传统的轴向、径向单一充磁对比,Halbach充磁的磁路方案磁力更大;新型的非完整式磁布置方式比传统的对称布置方案在竖直间隙为0.05 mm时提高磁力17倍,在竖直间隙为0.3 mm时提高磁力40倍;当轴颈与轴承孔间隙大于2.025 mm时,磁力占主导,可用于低速时脱开摩擦副,减少接触摩擦磨损;当轴颈与轴承孔间隙小于2.0216 mm时,水膜力逐渐增大,磁力起到减载作用;在工作间隙下,水膜力刚度系数比磁力刚度系数大2~3个数量级。磁液双浮轴承为提高纯磁轴承的稳定性提供了新的解决思路。

关键词：船舶水润滑尾轴承磁液双浮承载能力有限元法 Halbach阵列

动静压轴承支承电主轴服役精度保持用磁力减载研究

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振动与冲击 2017年第12期36卷 25-30页

作者：陈润霖欧阳武王建磊袁小阳西安交通大学现代设计及转子轴承系统教育部重点实验室西安710049 武汉理工大学能源与动力工程学院武汉430063 西安理工大学机械与精密仪器工程学院西安710048

为了保持动静压轴承支承电主轴的服役精度,提出采用电磁力消减静动态加工力的方案并研制了一套磁力减载系统。以磨床为例,建立电主轴系统的动力学模型,获取了载荷与轴端位移的关系;估算了磨削加工时产生的静态和动态加工力,提出了服役... 详细信息

为了保持动静压轴承支承电主轴的服役精度,提出采用电磁力消减静动态加工力的方案并研制了一套磁力减载系统。以磨床为例,建立电主轴系统的动力学模型,获取了载荷与轴端位移的关系;估算了磨削加工时产生的静态和动态加工力,提出了服役精度保持的静/动载荷需求;设计并建造了磁力减载系统,包括结构和测控两个模块;其中,结构模块特色在于磁力减载组件配备有万向支撑方案,测控模块支持开环和闭环两种控制策略;设计专项试验验证了减载系统的性能。研究结果表明:不转时减载组件可实现约35 N静载和28 N幅值谐波动载功能,初步证明采用电磁技术消减机床加工力是可行的。针对引入减载装置的电主轴系统,采用人工调节电流的开环控制策略更有效。

关键词：动静压轴承电主轴磁力减载精度保持服役精度