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A Review of Measurement-Integrated Simulation of Complex Real Flows
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作者 Toshiyuki Hayase 《Journal of Flow Control, Measurement & Visualization》 2015年第2期51-66,共16页
In spite of the inherent difficulty, reproducing the exact structure of real flows is a critically important issue in many fields, such as weather forecasting or feedback flow control. In order to obtain information o... In spite of the inherent difficulty, reproducing the exact structure of real flows is a critically important issue in many fields, such as weather forecasting or feedback flow control. In order to obtain information on real flows, extensive studies have been carried out on methodology to integrate measurement and simulation, for example, the four-dimensional variational data assimilation method (4D-Var) or the state estimator such as the Kalman filter or the state observer. Measurement-integrated (MI) simulation is a state observer in which a computational fluid dynamics (CFD) scheme is used as a mathematical model of the physical system instead of a small dimensional linear dynamical system usually used in state observers. A large dimensional nonlinear CFD model makes it possible to accurately reproduce real flows for properly designed feedback signals. This review article surveys the theoretical formulations and applications of MI simulation. Formulations of MI simulation are presented, including governing equations of a flow field observer, those of a linearized error dynamics describing the convergence of the observer, and stabilization of the numerical scheme, which is important in implementation of MI simulation. Applications of MI simulation are presented ranging from fundamental turbulent flows in pipes and Karman vortices in a wind tunnel to clinical application in diagnosis of blood flows in a human body. 展开更多
关键词 real flow Field State OBSERVER NUMERICAL SIMULATION MEASUREMENT Measurement-Integrated SIMULATION
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Gas Flow in Unilateral Opening Pulse Tubes Based on Real Gas Equation of State 被引量:3
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作者 代玉强 胡大鹏 +1 位作者 邹久朋 赵家权 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2009年第6期914-918,共5页
The real gas effect is dominant at high pressure and low temperature, and it is modeled by complex equations of state other than perfect gas law. In the vicinity of liquid-vapor critical point, the real gas exhibits u... The real gas effect is dominant at high pressure and low temperature, and it is modeled by complex equations of state other than perfect gas law. In the vicinity of liquid-vapor critical point, the real gas exhibits unusual gas dynamic behavior. In the present work, a transient wave fields in unilateral opening pulse tube is simulated by solving the Navier-Stokes equations incorporated with the Peng-Robinson thermodynamic model. The computational fluid dynamics (CFD) results show a remarkable deviation between perfect gas model and real gas model for contact interface and shockwave. The wave diagram based on the real gas model can help to solve the problem of offset design point. 展开更多
关键词 real gas flow pulse tube expansion refrigeration unsteady flow
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Remarks on a Real Viscous Heat-conducting Flow with Shear Viscosity
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作者 ZHANGZhi-ping WANGXiu-qin DENGShu-xian 《Chinese Quarterly Journal of Mathematics》 CSCD 北大核心 2005年第1期85-89,共5页
This paper is concerned with the global existence and exponential stability of solutions with large initial date in H^1 for real viscous heat-conducting flow with shear viscosity.
关键词 real viscous flow shear viscosity global existence
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Real-time Three-Dimensional Color Doppler Flow Imaging: An Improved Technique for Quantitative Analysis of Aortic Regurgitation 被引量:3
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作者 吕清 刘夏天 +3 位作者 谢明星 王新房 王静 庄磊 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2006年第1期148-152,共5页
The recently introduced real-time three-dimensional color Doppler flow imaging (RT-3D CDFI) technique provides a quick and accurate calculation of regurgitant jet volume (RJV) and fraction. In order to evaluate RT... The recently introduced real-time three-dimensional color Doppler flow imaging (RT-3D CDFI) technique provides a quick and accurate calculation of regurgitant jet volume (RJV) and fraction. In order to evaluate RT-3D CDFI in the noninvasive assessment of aortic RJV and regurgitant jet fraction (RJF) in patients with isolated aortic regurgitation, real-time three-dimensional echocardiographic studies were performed on 23 patients with isolated aortic regurgitation to obtain LV end-diastolic volumes (LVEDV), end-systolic volumes (LVESV) and RJV, and then RJF could be calculated. The regurgitant volume (RV) and regurgitant fraction (RF) calculated by two-dimensional pulsed Doppler (2D-PD) method served as reference values. The results showed that aortic RJV measured by the RT-3D CDFI method showed a good correlation with the 2D-PD measurements (r= 0.93, Y=0.89X+ 3.9, SEE= 8.6 mL, P〈0.001 ); the mean (SD) difference between the two methods was - 1.5 (9.8) mL. % RJF estimated by the RT-3D CDFI method was also correlated well with the values obtained by the 2D-PD method (r=0.88, Y=0.71X+ 14.8, SEE= 6.4 %, P〈0. 001); the mean (SD) difference between the two methods was -1.2 (7.9) %. It was suggested that the newly developed RT-3D CDFI technique was feasible in the majority of patients. In patients with eccentric aortic regurgitation, this new modality provides additional information to that obtained from the two-dimensional examination, which overcomes the inherent limitations of two-dimensional echocardiography by depicting the full extent of the jet trajectory. In addition, the RT-3D CDFI method is quick and accurate in calculating RJV and RJF. 展开更多
关键词 real-time three-dimensional echocardiography color Doppler flow imaging aortic regurgitation
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Analysing Traffic Flow and Traffic Hotspots from Historic and Real-Time GPS Data
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作者 Christopher Bartolo Thiago Matos Pinto 《通讯和计算机(中英文版)》 2015年第6期318-325,共8页
关键词 交通流分析 数据分析 历史 实时 数据采集方法 全球定位系统 道路网络 数据收集
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面向边缘计算的新能源台区实数化潮流算法
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作者 陈谦 冯源 +2 位作者 李成 徐旸 王姝莼 《电气传动》 2025年第1期61-69,共9页
供电台区接入的新能源电源比例日益提高,不可回避地带来了调控需求,因此亟需相适应的边缘潮流算法。为了在通信条件薄弱、计算资源有限的场景下,兼顾准确性、实用性、高效性,提出一种面向400 V供电台区边缘计算的实数化潮流算法。鉴于40... 供电台区接入的新能源电源比例日益提高,不可回避地带来了调控需求,因此亟需相适应的边缘潮流算法。为了在通信条件薄弱、计算资源有限的场景下,兼顾准确性、实用性、高效性,提出一种面向400 V供电台区边缘计算的实数化潮流算法。鉴于400 V台区网络呈阻性且设备功率因数较高的特点,将交流变量从向量和简化为标量和;设计了相应的节点转化策略与迭代算法;提出了数据缺失情况下的赋值策略;进一步提出了基于灵敏度修正或再次迭代计算的在线计算策略。通过仿真算例分析,最后验证了所提算法的准确性与实用性。 展开更多
关键词 边缘计算 新能源台区 实数化 潮流算法 计算策略
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单分子实时测序在α珠蛋白基因三联体及其复合变异等位基因鉴定中的临床应用
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作者 张宇 方艳平 +3 位作者 朱碧青 梁丽仪 周万军 江凌晓 《国际检验医学杂志》 2025年第1期32-37,43,共7页
目的探究单分子实时测序(SMRT)技术在α珠蛋白基因三联体(简称α三联体)及其复合变异等位基因鉴定中的临床应用效果。方法收集α三联体阳性样本36例,其中经PCR-导流杂交技术确认28例、经高通量测序(NGS)技术确认8例。36例样本包含α三... 目的探究单分子实时测序(SMRT)技术在α珠蛋白基因三联体(简称α三联体)及其复合变异等位基因鉴定中的临床应用效果。方法收集α三联体阳性样本36例,其中经PCR-导流杂交技术确认28例、经高通量测序(NGS)技术确认8例。36例样本包含α三联体复合变异顺式或反式排列未明样本ααα^(anti4.2)复合α^(CS)α2例,ααα^(anti4.2)复合-α^(3.7)10例,HKαα/--SEA型待确证2例,均采用SMRT技术进行地中海贫血(简称地贫)基因检测。另招募一个基因型为ααα^(anti4.2)复合-α^(3.7)变异病例的家系,包含先证者(Ⅱ-1)和其父亲(Ⅰ-1)、母亲(Ⅰ-2),采用PCR-导流杂交和SMRT技术进行地贫基因检测。结果SMRT技术检测结果显示,36例样本中共检出35例α三联体,1例αααα^(anti4.2)型α珠蛋白基因四联体(简称α四联体)。2例ααα^(anti4.2)复合α^(CS)α变异样本中ααα^(anti4.2)与α^(CS)α均为反式排列,基因型为ααα^(anti4.2)/α^(CS)α,10例ααα^(anti4.2)复合-α^(3.7)变异样本中ααα^(anti4.2)与-α^(3.7)为顺式排列样本9例,基因型为HKαα/αα,ααα^(anti4.2)与-α^(3.7)为反式排列样本1例,基因型为ααα^(anti4.2)/-α^(3.7)。相较PCR-导流杂交技术,SMRT技术多检出1例大片段缺失型β珠蛋白基因变异和2例未知变异,阳性检出率提高10.71%(3/28)。家系分析表明,先证者(Ⅱ-1)ααα^(anti4.2)和-α^(3.7)变异均遗传自母亲(Ⅰ-2),其基因型为HKαα/αα,与SMRT技术检测结果一致。结论SMRT技术不仅能够准确检测α三联体、α四联体及其复合变异等位基因,而且具有高准确性、一步到位识别2种变异顺式或反式排列、地贫基因变异覆盖全等优势,具有良好的临床应用价值。 展开更多
关键词 α珠蛋白基因三联体 HKαα 单分子实时测序 PCR-导流杂交技术 高通量测序
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Mechanisms for non-ideal flow in low-power arc-heated supersonic nozzles 被引量:1
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作者 Cheng-Kang Wu Wen-Xia Pan +1 位作者 Xian Meng Hai-Xing Wang 《Acta Mechanica Sinica》 SCIE EI CAS CSCD 2015年第4期500-511,共12页
The flow in a low-powered arc gas heater com- bined with a supersonic nozzle of throat diameter less than 1 mm is quite complicated and difficult to describe in quan- titative detail. Experiments on arc-heated superso... The flow in a low-powered arc gas heater com- bined with a supersonic nozzle of throat diameter less than 1 mm is quite complicated and difficult to describe in quan- titative detail. Experiments on arc-heated supersonic jet thrusters of monatomic gases argon and helium have been carried out and their performance measured. The flow charac- teristics are analyzed with the help of numerical simulation. Results show that the viscous effect is the most important factor causing the large difference between ideal and real performance. A large outer section of the exit flow is slow- moving. This is especially pronounced in helium, where 70 % of the exit area of the nozzle might be in subsonic flow. Fric- tion forces can be much larger than the net thrust, reaching several times higher in helium, resulting in very low efficien- cies. Other factors causing the differences between ideal and real flow include: complex flow in the throat region, electric arc extending to the nozzle expansion section, heat transfer to the inlet gas and from the hot plasma, and environmen- tal pressure in the vacuum chamber. It is recognized that the ordinary concepts of supersonic nozzle flow must be greatly modified when dealing with such complicated situations. The general concepts presented in this paper could be helpful in guiding the design and operation of this equipment. 展开更多
关键词 Arc-heated supersonic jet LOW-POWER real flow Viscosity effect Low Reynolds number
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采用Netflow数据的典型异常流量检测方法 被引量:5
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作者 田杨 王宏 陈晓梅 《电子科技大学学报》 EI CAS CSCD 北大核心 2009年第S1期57-60,74,共5页
基于Netflow数据提出了根据流量特征进行异常检测的方法;分析了造成异常流量的DDoS和端口扫描的流量特征两种网络攻击行为;并根据其特征进行用户可控的实时异常流量检测,给出告警,报告异常的时空坐标。用户可以调整自己的参数设置,在计... 基于Netflow数据提出了根据流量特征进行异常检测的方法;分析了造成异常流量的DDoS和端口扫描的流量特征两种网络攻击行为;并根据其特征进行用户可控的实时异常流量检测,给出告警,报告异常的时空坐标。用户可以调整自己的参数设置,在计算时间和空间上平衡自己的参数,得到满意的结果。采用Web形式和CS架构模式进行异常监控的实时显示,用户可以实时地在任何连接到服务器的主机设置参数和查看检测结果。 展开更多
关键词 DDOS 流量特征 NETflow 端口扫描 实时检测
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Intercross real-time control strategy in alternating activated sludge process for short-cut biological nitrogen removal treating domestic wastewater 被引量:4
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作者 AKIO TAKIGAWA 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2008年第8期957-963,共7页
To develop technically feasible and economically favorable dynamic process control(DPC)strategies for an alternating activated sludge(AAS)system,a bench-scale continuous-flow alternating aerobic and anoxic reactor,per... To develop technically feasible and economically favorable dynamic process control(DPC)strategies for an alternating activated sludge(AAS)system,a bench-scale continuous-flow alternating aerobic and anoxic reactor,performing short-cut nitrogen removal from real domestic wastewater was operated under different control strategies for more than five months.A fixed-time control(FTC) study showed that bending-points on pH and oxidation-reduction potential(ORP)profiles accurately coincided with the major biologic... 展开更多
关键词 alternating activated sludge process CONTINUOUS-flow real-time control NITRIFICATION
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Real-Time Traffic Signal Timing for Urban Road Multi-Intersection
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作者 Lin Dong Wushan Chen 《Intelligent Information Management》 2010年第8期483-486,共4页
this paper develops a real-time traffic signal timing model which is to be integrated into a single intersection for urban road, thereby solving the problem of traffic congestion. We analyze the current situation of t... this paper develops a real-time traffic signal timing model which is to be integrated into a single intersection for urban road, thereby solving the problem of traffic congestion. We analyze the current situation of the traffic flow with release matrix firstly, and then put forward the basic models to minimize total delay time of vehicles at the intersection. The optimal real-time signal timing model (non-fixed cycle and non-fixed split) is built with the Webster split optimal model. At last, the simulated results, which are compared with conventional model, manifest the promising properties of proposed model. 展开更多
关键词 TRAFFIC SIGNAL Control TRAFFIC flows real-Time SIGNAL Timing Release Matrix SPLIT PASSION Distribution
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基于能量守恒定律对涡轮流量计的脉动误差上限的估算
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作者 刘可薇 胡银春 张禾 《电子测量与仪器学报》 CSCD 北大核心 2024年第2期85-91,共7页
在管道输运当中,非稳态流会使涡轮流量计的计量结果产生偏差。为了指导脉动流工况下的涡轮流量计选型和修正,需估算脉动工况对涡轮流量计产生的最大脉动误差。忽略流体压力和温度变化影响,基于涡轮流量计的动态特性和能量守恒定律,在一... 在管道输运当中,非稳态流会使涡轮流量计的计量结果产生偏差。为了指导脉动流工况下的涡轮流量计选型和修正,需估算脉动工况对涡轮流量计产生的最大脉动误差。忽略流体压力和温度变化影响,基于涡轮流量计的动态特性和能量守恒定律,在一个脉动周期内计算脉动流和定常流对叶轮的做功比值,推导得出正弦脉动流误差上限公式。基于6DOF模型和UDF建立了涡轮流量计CFD仿真模型,搭建了空气流实验平台,以工作级流量标准装置作为标准表,被校表和标准表间采用内置整流器的管段相连。对比定常流下的流量值,计算各脉动工况下的误差限值。综合各脉动工况点,上限公式与CFD仿真所得脉动误差上限的最大差值为0.281%,与空气流实验所得脉动误差上限的最大差值为0.224%。经CFD仿真和空气流实验结果对比验证,脉动误差上限公式较为准确,可以直接用于脉动频率大于10 Hz的工况下误差值的估计和修正。 展开更多
关键词 涡轮流量计 数学模型 CFD 正弦波脉动流 实流实验
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NEW MODEL OF GAS FLOW PROBLEM IN MULTI-LAYERED GAS RESERVOIR AND APPLICATION
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作者 李笑萍 《Applied Mathematics and Mechanics(English Edition)》 SCIE EI 1993年第12期1133-1141,共9页
In this paper, the new model of the real gas filtration problem has been presented multi-layered gas reservoir, when a gas well output and wellbore storage may be variable, and have obtained the exact solutions of pre... In this paper, the new model of the real gas filtration problem has been presented multi-layered gas reservoir, when a gas well output and wellbore storage may be variable, and have obtained the exact solutions of pressure distribution for each reservoir bed under three kinds of typical out-boundary conditions. As a special case, according to the new model have also obtained the qxact solutions of presssure distribution in homogeneous reservoir and is given important application in gas reservoir development. 展开更多
关键词 multi-layered gas reservoir gas flow model of real gas filtration
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A Simple Strategy for Capturing the Unstable Steady Solution of an Unsteady Flow
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作者 Weiwei Zhang Yilang Liu Jing Li 《Open Journal of Fluid Dynamics》 2015年第2期188-198,共11页
A simple strategy for capturing unstable steady solution is proposed in this paper. By enlarging the real time step, the unstable high frequency modes of the unsteady flow can be effectively suppressed because of the ... A simple strategy for capturing unstable steady solution is proposed in this paper. By enlarging the real time step, the unstable high frequency modes of the unsteady flow can be effectively suppressed because of the damping action on the real time level. The steady component will not be changed during real time marching. When the unsteady flow solution converges to a steady state, the partial derivatives of real time in unsteady governing equations will disappear automatically. The steady solution is the exacted unstable steady solution. The method is validated by the laminar flow past a circular cylinder and a transonic buffet of NACA0012 airfoil. This approach can acquire high-precision unstable steady solutions quickly and effectively without reducing the spatial discretization accuracy. This work provides the basis for unstable flow modeling and analysis. 展开更多
关键词 UNSTABLE STEADY SOLUTION real Time STEP Modeling UNSTEADY flowS
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远郊大型冬季活动客流演进及路径分流研究
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作者 秦焕美 贾美茹 +2 位作者 钱翠 曹静 陈艳艳 《重庆交通大学学报(自然科学版)》 CAS CSCD 北大核心 2024年第7期85-93,共9页
有效的客流组织是保证大型活动成功举办的关键,通过大型冬季活动的客流特征及意向调查,建立了观众路径选择模型和客流动态演进模型,分析了大型活动各设施节点的客流分布和客流排队时间分布动态变化规律。结果表明:增加取暖房、纪念品店... 有效的客流组织是保证大型活动成功举办的关键,通过大型冬季活动的客流特征及意向调查,建立了观众路径选择模型和客流动态演进模型,分析了大型活动各设施节点的客流分布和客流排队时间分布动态变化规律。结果表明:增加取暖房、纪念品店、景观点等设施将提升观众在远郊大型冬季活动中的观赛体验。大型冬季活动的观众可接受的安检/检票、等公交的排队时间及步行到场馆的时间心理阈值为20 min,且观众更愿意选择步行时间短和服务设施数量多的路径;采用路径分流并提供路径实时信息,会使得分流路径上、下游节点的客流分布更为均衡平缓,有效减少客流聚集和长时间排队等待现象。 展开更多
关键词 交通工程 远郊大型冬季活动 客流动态演进 路径分流 实时信息 客流分布
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基于YOLO模型的车流量实时采集系统研究
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作者 王金环 李宝敏 《计算机技术与发展》 2024年第9期209-214,共6页
对于一座现代化城市来说,合理的交通规划是一个城市高效运行的关键,作为交通规划的关键信息的城市车流量信息,原本需要人工进行识别、获取、验证的提取方式,随着计算机视觉技术的蓬勃发展弊端尽显,终将退出历史的舞台。为了提高城市车... 对于一座现代化城市来说,合理的交通规划是一个城市高效运行的关键,作为交通规划的关键信息的城市车流量信息,原本需要人工进行识别、获取、验证的提取方式,随着计算机视觉技术的蓬勃发展弊端尽显,终将退出历史的舞台。为了提高城市车流量信息的准确性和及时性,利用现有的计算机技术设计一种基于YOLO模型的车流量实时采集系统。该系统基于YOLO视觉检测模型,采用DeepSORT算法对检测到的目标车辆进行跟踪识别、判断车辆的运行状态、实现当前路段的车流量统计、对已记录车流量信息进行可视化展示以及数据输出等。该系统可以有效地代替传统消耗人力的死板工作,实现自动化数据收集以及道路交通情况的快速监测。该系统操作简单,交互性强,为城市的交通管理和交通规划提供准确实时的信息数据。 展开更多
关键词 目标检测 目标跟踪算法 数据处理 YOLO模型 车流量 实时采集
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基于实时交通流的上海市机动车碳污同源排放清单
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作者 刘启贞 巢渊 +5 位作者 易敏 王磊 刘登国 刘娟 朱洪 伏晴艳 《上海计量测试》 2024年第2期59-62,68,共5页
为满足交通碳污减排政策多样化、差别化和精细化发展需求,扩大交通排放测算范围、提高测算准确度和时空分辨力,提出了一种基于实时交通流的高时空分辨力机动车碳污同源排放清单构建方法,综合交通模型、IVE模型和相关算法建立了覆盖上海... 为满足交通碳污减排政策多样化、差别化和精细化发展需求,扩大交通排放测算范围、提高测算准确度和时空分辨力,提出了一种基于实时交通流的高时空分辨力机动车碳污同源排放清单构建方法,综合交通模型、IVE模型和相关算法建立了覆盖上海全市的机动车碳污同源排放清单。结果显示,二氧化碳、氮氧化物周排放规律相近,呈现出明显的周末低工作日高的特征;二氧化碳分时变化规律呈现明显的双峰,而氮氧化物受重型车影响,峰值出现在夜间;由于各车型的碳污分担率不同且活动区域不同造成了上海市机动车二氧化碳和氮氧化物排放的空间分布差异明显;碳污同源排放清单中,NOx、PM、CO、VOCs、CO_(2)的月均排放量分别为5160t、440t、1.7×10^(4)t、732t和2.46×10^(6)t;排放分担率表明,NOx、PM等污染物主要来自重货车及大货车排放;CO、VOCs和CO_(2)排放来源以小客车排放占比最高。 展开更多
关键词 交通环境保护 机动车排放清单 实时交通流 动态排放清单 大气污染治理
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企业负债率对实体企业金融化的中介效应与门槛效应——基于债务税盾角度
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作者 陈丽娜 马深隆 杨雪 《科技和产业》 2024年第19期49-58,共10页
以2010—2022年中国A股上市公司为研究样本,基于债务税盾视角,选取企业现金流为中介变量,检验企业负债率对实体企业金融化的影响。实证结果表明:企业负债率与实体企业金融化显著负相关;企业现金流在企业负债率对实体企业金融化的影响过... 以2010—2022年中国A股上市公司为研究样本,基于债务税盾视角,选取企业现金流为中介变量,检验企业负债率对实体企业金融化的影响。实证结果表明:企业负债率与实体企业金融化显著负相关;企业现金流在企业负债率对实体企业金融化的影响过程中起部分中介作用;企业负债率对实体企业金融化存在门槛效应,当企业负债率超过门槛值时,企业负债率对实体企业金融化的抑制作用明显减弱;代理问题会削弱企业负债率对实体企业金融化的抑制作用;相较于民营企业,企业负债率对实体企业金融化的负向影响在国有企业中更加明显。 展开更多
关键词 企业负债率 实体企业金融化 企业现金流 债务税盾 中介效应 门槛效应
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基于RPA的病原体快速诊断策略 被引量:3
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作者 许淑莹 王冬梅 欧阳松应 《福建师范大学学报(自然科学版)》 CAS 北大核心 2024年第1期34-44,共11页
重组酶聚合酶扩增(recombinase enzyme polymerase amplification,RPA)检测技术是2006年出现的一种等温核酸扩增检测技术。RPA所需引物设计简单,反应时间短,在37~42℃下仅需15~20 min即可达到病原体检测水平,搭配便捷装置即可实现核酸检... 重组酶聚合酶扩增(recombinase enzyme polymerase amplification,RPA)检测技术是2006年出现的一种等温核酸扩增检测技术。RPA所需引物设计简单,反应时间短,在37~42℃下仅需15~20 min即可达到病原体检测水平,搭配便捷装置即可实现核酸检测,近些年得到快速发展。重点介绍了RPA反应产物检测的相关策略,并对RPA在致病菌、病毒以及寄生虫上的快速诊断应用研究进行述评。 展开更多
关键词 等温扩增 重组酶聚合酶扩增 侧向流动 实时荧光检测 病原体检测
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基于多交叉置换扩增和纳米生物传感技术快速检测肺炎支原体方法的建立
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作者 肖飞 郑宝英 +8 位作者 徐文健 伏瑾 黄小兰 孙春荣 贾楠 张裕 许峥 周娟 王毅 《遵义医科大学学报》 2024年第5期513-521,共9页
目的建立一种简单、灵敏、快速的肺炎支原体(MP)检测方法,并对其应用性进行验证和评价。方法利用多交叉置换扩增(MCDA)技术对肺炎支原体特异基因CARDS毒素基因进行扩增,利用侧流免疫层析生物传感(LFB)技术读取扩增结果,命名该方法为MP-M... 目的建立一种简单、灵敏、快速的肺炎支原体(MP)检测方法,并对其应用性进行验证和评价。方法利用多交叉置换扩增(MCDA)技术对肺炎支原体特异基因CARDS毒素基因进行扩增,利用侧流免疫层析生物传感(LFB)技术读取扩增结果,命名该方法为MP-MCDA-LFB。分析扩增反应在60~67℃(间隔1℃)的扩增效率,筛选最适反应温度;分析分别扩增10、20、30、40 min时能够检测到的最低核酸浓度,筛选最佳反应时间。利用10倍系列稀释的肺炎支原体核酸分析MP-MCDA-LFB方法的灵敏度和检测限,利用35株非肺炎支原体菌株分析MP-MCDA-LFB方法的特异性。利用MP-MCDA-LFB方法检测80份疑似MP感染的临床样本,并与RT-PCR法检测结果进行比较,分析MP-MCDA-LFB方法的临床应用性。结果MP-MCDA-LFB能够实现对肺炎支原体CARDS毒素基因的快速检测。其最佳反应温度为63℃,最短反应时间为40 min,整个检测过程可在1 h内。MP-MCDA-LFB方法具有较高的灵敏度和特异性,其检测限低至45 ng/L,与其他临床表现相似的病原体无交叉反应,特异性为100%。MP-MCDA-LFB方法从80份临床样本中检出45份阳性样本(56.3%),检出率与RT-PCR方法一致。结论本研究建立的以CARDS毒素基因为靶标的MP-MCDA-LFB检测方法具有简单、快速、灵敏度高、特异性强的优点,在基层医疗机构和现场检测具有较好的应用潜力。 展开更多
关键词 肺炎支原体 多交叉置换扩增技术 侧流免疫层析生物传感技术 CARDS毒素基因 RT-PCR
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