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敲除LTA4H基因的PK-15细胞系构建及其对口蹄疫病毒复制的影响

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微生物学报 2024年第3期64卷 733-744页

作者：王家丽蒲秀瑛杨帆邵文华黄梦瑶曹伟军郑海学张伟兰州理工大学生命科学与工程学院甘肃兰州730050 中国农业科学院兰州兽医研究所/兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室甘肃兰州730000 甘肃省病原生物学基础学科研究中心甘肃兰州730046

【目的】利用CRISPR/Cas9基因编辑技术构建白三烯A4水解酶(leukotriene A4 hydrolase,LTA4H)基因缺失的猪肾细胞系(porcine kidney-15,PK-15),探究LTA4H对口蹄疫病毒(foot-and-mouth disease virus,FMDV)复制的影响,为开展LTA4H功能研究及调控病毒复制机制研究提供理论依据。【方法】设计2条针对猪LTA4H基因的引导RNA(small guide RNA,sgRNA),分别构建至载体pX459-puro-MCS中;将CRISPR重组质粒转染PK-15细胞,用嘌呤霉素(puromycin)抗生素筛选,并通过有限稀释法筛选单克隆细胞,之后通过Western blotting和测序检测LTA4H基因的敲除,获得LTA4H基因功能缺失细胞系。使用Western blotting、RT-qPCR及病毒滴度测定等方法检测敲除LTA4H基因后对FMDV复制及相关蛋白的表达情况。【结果】获得的单克隆敲除细胞系与野生型细胞相比,能够显著抑制FMDV复制。【结论】本研究成功构建了LTA4H基因敲除的PK-15细胞系,证明LTA4H对FMDV的复制具有促进作用,研究结果为后续LTA4H功能研究提供理论依据。

关键词： CRISPR/Cas9 白三烯A4水解酶细胞系口蹄疫病毒

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Tollip敲除猪肾细胞系的构建

畜牧兽医学报 2024年第4期55卷 1810-1818页

作者：王家丽杨帆邵文华黄梦瑶曹伟军蒲秀瑛张伟郑海学中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室国家口蹄疫参考实验室兰州730000 兰州理工大学生命科学与工程学院兰州730050 甘肃省病原生物学基础学科研究中心兰州730046

旨在探究Toll作用蛋白(toll-interacting protein, Tollip)对口蹄疫病毒(foot-and-mouth disease virus, FMDV)增殖的影响,利用CRISPR/Cas9基因编辑技术构建Tollip基因缺失的猪肾细胞系(porcine kidney cell line, PK-15),并通过PCR测序和Western blot确认敲除效果,然后利用CCK-8试剂盒测定Tollip缺失后细胞活力变化,确认敲除细胞与野生细胞无显著差异,用FMDV感染敲除细胞后,采用Western blot、间接免疫荧光、实时荧光定量和病毒滴度测定等方法测定病毒在敲除细胞系中复制情况。结果显示:感染FMDV后,敲除细胞与野生细胞相比,显著促进了病毒的复制。综上,在PK-15细胞上,成功构建Tollip缺失细胞系,且试验表明Tollip的缺失有助于FMDV的复制。研究结果为后续Tollip功能研究及抗病毒机制研究提供理论依据。

关键词： CRISPR/Cas9 Tollip基因细胞系口蹄疫病毒

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口蹄疫病毒非结构蛋白3A的研究进展

中国兽医科学 2024年第4期54卷 441-445页

作者：王家丽杨帆邵文华黄梦瑶曹伟军张伟郑海学中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室甘肃兰州730046 兰州理工大学生命科学与工程学院甘肃兰州730050 甘肃省病原生物学基础学科研究中心甘肃兰州730046

口蹄疫(FMD)是由口蹄疫病毒(FMDV)引起的一种最具影响、最难控制的动物疫病之一,严重危害着偶蹄动物的健康。FMDV属于单股正链RNA病毒,由结构蛋白和非结构蛋白组成。本文系统总结了口蹄疫病毒非结构蛋白3A的结构和功能特点,有助于进一... 详细信息

口蹄疫(FMD)是由口蹄疫病毒(FMDV)引起的一种最具影响、最难控制的动物疫病之一,严重危害着偶蹄动物的健康。FMDV属于单股正链RNA病毒,由结构蛋白和非结构蛋白组成。本文系统总结了口蹄疫病毒非结构蛋白3A的结构和功能特点,有助于进一步研究FMDV的感染机制并为FMD防控提供参考。

关键词：口蹄疫病毒非结构蛋白 3A 结构功能

过表达JMJD6基因的293T细胞系的构建及对水疱性口炎病毒增殖效率的评价

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中国兽医科学 2024年第5期54卷 569-576页

作者：黄梦瑶杨帆杨洋邵文华王家丽吕岳芹赵晓义陈治彤曹伟军张伟郑海学中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室甘肃兰州730000 甘肃省病原生物学基础学科研究中心甘肃兰州730046

为提高水疱性口炎病毒(vesicular stomatitis virus,VSV)在细胞中的增殖效率,本研究利用慢病毒包装系统构建Jumonji C结构域蛋白6(Jumonji domain-containing protein 6,JMJD6)过表达的HEK-293T/JMJD6细胞系,分析VSV在HEK-293T/JMJD6细胞系与HEK-293T细胞中的复制差异。首先以JMJD6基因为目标,构建重组质粒pLV-puro-3×Flag-JMJD6,并与辅助质粒pMD2.G和psPAX2共转染至HEK-293T细胞进行慢病毒包装。将包装好的慢病毒感染HEK-293T细胞并通过嘌呤霉素筛选获得过表达JMJD6基因的阳性细胞。利用Western-blot、间接免疫荧光试验、流式细胞术和蚀斑试验检测VSV在HEK-293T/JMJD6细胞系中的复制能力。结果显示,在HEK-293T/JMJD6细胞系中,VSV的复制能力显著增强。实时荧光定量检测VSV感染的HEK-293T/JMJD6细胞系和野生型HEK-293T细胞中干扰素下游基因的表达,结果显示HEK-293T/JMJD6细胞系抑制VSV诱导的Ⅰ型干扰素下游基因的产生。总之,本研究构建的HEK-293T/JMJD6细胞系能显著促进VSV的增殖并抑制Ⅰ型干扰素信号通路,为VSV疫苗候选细胞株的筛选奠定了基础。

关键词： JMJD6 水疱性口炎病毒慢病毒包装过表达细胞系干扰素

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A Bluetooth Localization Algorithm Based on Map Path Calibration and Time Series Filtering

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A Bluetooth Localization Algorithm Based on Map Path Calibra...

A Robust Turn Detection Algorithm based on Periodic Signal Identification

第九届中国卫星导航学术年会

作者： shaomeng Chen Haiyong LUO Qu Wang wenhua shao Fang Zhao School of Software Engineering Beijing University of Posts and Telecommunications Research Centre of Pervasive Computing Institute of Computing Technology Chinese Academy of Sciences School of Information and Communication Engineering Beijing University of Posts and Telecommunications

With the gradual progress of the urbanization process of modern society and the continuous improvement of residents’ living standard, people’s demand for IPS(Indoor Positioning System) is more urgent than that of the past, higher requirements for positioning accuracy, positioning robustness and positioning energy consumption are put forward. Low-power Bluetooth technology has the characteristics of low-cost, easy deployment and low power consumption, supporting all mainstream smartphone operating systems, and attracting extensive attention from academic and industry field. In this paper, aiming at the situation that current Bluetooth low energy positioning accuracy has large fluctuation under the circumstance of multipath propagation, external disturbance and other factors, and has poor user experience, a Bluetooth localization algorithm based on map path calibration and time series filtering is proposed(MCTF). This algorithm comprehensively utilizes a post-processing method based on the average sliding window, time-series based spatial restraint post-processing method and path information based location calibration method, effectively eliminating the large fluctuation of positioning results caused by multipath propagation and other external wireless signal interference in complex indoor environment. By using map information, based on the fact that position cannot be transient, utilizing feedback filtering, positioning results can get path correction and directioncorrection respectively, solving the problem that the positioning coordinates jump between adjacent narrow aisles and jump back and forth on the same path during the positioning process. Extensive experiments performed in shopping mall, building office, exhibition center and other typical indoor environments demonstrate that the average positioning error of the proposed algorithm is within 1.5 m, which has good robustness and ensures the smoothness, accuracy and stability of the positioning result.

关键词： indoor positioning Bluetooth location map path information time series filtering BLE path correction direction correction

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A Robust Turn Detection Algorithm based on Periodic Signal I...

Topologically Correct Intersection Curves of Two Trimmed Quadrics with Tolerance Control

第九届中国卫星导航学术年会

作者： Yu Chen Haiyong Luo Fang Zhao wenhua shao Qu Wang School of Software Engineering Beijing University of Posts and Telecommunications Research Centre of Pervasive Computing Institute of Computing Technology Chinese Academy of Sciences School of Information and Communication Engineering Beijing University of Posts and Telecommunications

Turn detection can be widely used in location navigation, user behavior identification, and scene awareness. Accurate real-time identification of pedestrian turns contributes to improving the accuracy of positioning navigation accuracy and scene awareness. Considering the problem of the insufficient accuracy and robustness of the existing turn detection algorithm, this paper proposes a turn detection algorithm based on periodic signal recognition, which effectively solves the misjudgment caused by the periodic swing of user walking. The algorithm collects the acceleration sensor and the gyro sensor data in real time and calculates the angular velocity of the rotation in the vertical direction by multiplying the vertical gravitational acceleration unit vector by the gyro sensor data. In this paper, fast Fourier transform(FFT) is used to identify and eliminate periodic interfering signals generated by user walking, only keeping the non-periodic signal generated by user walking(such as turning). By integrating the vertical angular velocity data in the sliding window to obtain the angle change value of the user walking within a certain period of time, and the threshold value is compared to realize the accurate identification of the turning behavior. In order to adapt to the user’s turn size and the turn speed, the algorithm also proposes a corner detection method based on multi-sliding window. The experiment results show that the proposed algorithm has higher accuracy and lower power consumption than other algorithms which based on GPS and electronic compass. The algorithm can realize the accurate identification of the turning behavior of the mobile phone in various positions of the user, and has good robustness. The accuracy of turning recognition can reach 93% and the average power consumption is about 60 mW.

关键词： turn detection scene awareness signal recognition multi-sliding window high accuracy low power consumption

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Journal of Systems Science & Complexity 2024年第5期37卷 2207-2239页

作者： shao Wenbing CHEN Falai Department of Mathematics University of Science and Technology of ChinaHefei 230026China

Surface/surface intersection is a fundamental problem in Compute Aided Design and Geometric Modeling since it is essential to solid modeling,numerically controlled machining,feature recognition,computer animation,*** practical applications,quadric surfaces,which are the most basic type of surfaces,are typically bounded surfaces trimmed by a sequence of *** this paper,a robust algorithm is proposed for computing the intersection curve segments of two trimmed quadrics based on the parametric representation of the intersection curves of two *** proposed algorithm guarantees correct topology and ensures that the approximation errors of the end points of the intersection curve segments are less than a given *** error control is based on an effective solution to a set of polynomial inequality system using the root isolation *** examples are presented to validate the robustness and effectiveness of the proposed algorithm.

关键词： Polynomial inequality system root isolation surface/surface intersection tolerance control trimmed quadrics

Pseudoeffective thresholds and cohomology of twisted symmetric tensor fields on irreducible Hermitian symmetric spaces

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Science China Mathematics 2024年第11期67卷 2433-2452页

作者： Feng shao Institute of Mathematics Academy of Mathematics and Systems ScienceChinese Academy of Sciences School of Mathematical Sciences University of Chinese Academy of Sciences

Let X be an irreducible Hermitian symmetric space of compact type(IHSS for short).In this paper,we give the irreducible decomposition of *** a by-product,we give a cohomological characterization of the rank of ***,we introduce pseudoeffective thresholds to measure the bigness of tangent bundles of smooth complex projective varieties precisely and calculate them for irreducible Hermitian symmetric spaces of compact type.

关键词： tangent bundle pseudoeffective threshold irreducible Hermitian symmetric space of compact type (IHSS) multiplicity free action

Homogeneous void nucleation in the presence of supersaturated vacancies and interstitials

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Journal of Materiomics 2024年第5期10卷 1147-1159页

作者： Lin shao Accelerator Laboratory Department of Nuclear EngineeringTexas A&M UniversityCollege StationTX77843USA

Homogeneous void nucleation in metals containing arbitrary vacancies and interstitials has been reexamined,with corrections made to the original work by Katz and *** void size distributions derived previously missed an exponential modification function with void size as the *** a result,void nucleation under a given vacancy supersaturation ratio is underestimated by orders of *** second improvement arises from the accuracy in calculating the vacancy arrival rate to a *** present work proposes establishing a direct relationship between the vacancy arrival rate and the available self-diffusion *** these corrections and improvements,void nucleation in pure Fe is calculated as an example,and an analytic fitting formula is *** required vacancy supersaturation ratio and interstitial-to-vacancy flux ratio for void nucleation calculations can be easily obtained from an analytical solution of rate theory calculations,in which dislocation density and displacements per atom(dpa)rate are adjustable ***,the nucleation rate calculation can be incorporated into rate theory calculations considering evolving dislocation densities,which leads to time-dependent void nucleation.

关键词： Void nucleation Voids Irradiation Swelling Defects