4种植物源性成分多重real-time PCR检测方法的建立及其在食用淀粉中的应用

建立一种可同时快速检测红薯、木薯、马铃薯、玉米源性成分的多重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以红薯g3pdh基因、木薯g3pdh基因、马铃薯UGPase基因、玉米zSSIIb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,开展方法学验证,并对不同掺入比例模拟样品和实际淀粉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。与15种非目标源性均无交叉反应;对目标DNA的检测灵敏度可达到3×10^(-3) ng/μL,且具有良好的线性关系和扩增效率;对淀粉样品的检出限可达0.1%,对50份实际样品进行检测,结果与参比方法一致,说明建立的多重real-time PCR法可用于食用淀粉种类掺假鉴别检测。

Real-time Rescue Target Detection Based on UAV Imagery for Flood Emergency Response

Timely acquisition of rescue target information is critical for emergency response after a flood disaster.Unmanned Aerial Vehicles(UAVs)equipped with remote sensing capabilities offer distinct advantages,including high-resolution imagery and exceptional mobility,making them well suited for monitoring flood extent and identifying rescue targets during floods.However,there are some challenges in interpreting rescue information in real time from flood images captured by UAVs,such as the complexity of the scenarios of UAV images,the lack of flood rescue target detection datasets and the limited real-time processing capabilities of the airborne on-board platform.Thus,we propose a real-time rescue target detection method for UAVs that is capable of efficiently delineating flood extent and identifying rescue targets(i.e.,pedestrians and vehicles trapped by floods).The proposed method achieves real-time rescue information extraction for UAV platforms by lightweight processing and fusion of flood extent extraction model and target detection model.The flood inundation range is extracted by the proposed method in real time and detects targets such as people and vehicles to be rescued based on this layer.Our experimental results demonstrate that the Intersection over Union(IoU)for flood water extraction reaches an impressive 80%,and the IoU for real-time flood water extraction stands at a commendable 76.4%.The information on flood stricken targets extracted by this method in real time can be used for flood emergency rescue.

基于病菌孢子捕捉和real-time PCR技术的田间空气中小麦白粉病菌孢子动态监测及病情估计模型研究

利用Burkard定容式孢子捕捉器结合real-time PCR定量技术,分别对种植高抗、中感和高感白粉病小麦品种的田间空气中白粉病菌分生孢子浓度进行监测,结果表明,real-time PCR定量与传统的显微观察计数两种方法测得的孢子浓度呈显著正相关(P≤0.01),且两种病菌孢子计数方法在同一抗性品种上监测到的孢子浓度动态相近。此外,两种方法测得的孢子浓度与各气象因子的相关性分析结果一致,空气中的白粉病菌孢子浓度主要与空气相对湿度显著正相关。在此基础上,利用两种方法测定的田间空气中白粉病菌孢子浓度分别建立了基于累积孢子浓度的田间病情估计模型。分析发现,基于两种孢子浓度测定方法建立的病情估计模型间无显著性差异,表明real-time PCR定量技术测定的孢子浓度在构建白粉病病情估计模型上具有一定可行性。该结果为real-time PCR定量技术与病菌孢子捕捉技术相结合用于小麦白粉病的监测和预测提供理论依据。

马乳酒样乳杆菌马乳酒样亚种real-time PCR检测方法的建立与应用

本研究建立了一种特异性实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)检测方法,根据模式菌株马乳酒样乳杆菌马乳酒样亚种ZW3的16S rDNA序列和全基因组序列设计筛选特异性引物,采用SYBR Green I荧光染料建立real-time PCR方法,并对方法的特异性、灵敏度、重复性和混合体系等进行检测。结果表明,本研究所建立的方法特异性强、灵敏度高、重复性好,建立real-time PCR的标准曲线,其决定系数R2为0.965,具有良好的线性关系,且在马乳酒样乳杆菌马乳酒样亚种及混合体系中可以特异性检出。综上,本研究建立的real-time PCR法可以快速、准确地检测马乳酒样乳杆菌马乳酒样亚种,为马乳酒样乳杆菌的特异性定性定量检测提供了一种新的方法。

Prescribed Performance Tracking Control of Time-Delay Nonlinear Systems With Output Constraints

The problem of prescribed performance tracking control for unknown time-delay nonlinear systems subject to output constraints is dealt with in this paper. In contrast with related works, only the most fundamental requirements, i.e., boundedness and the local Lipschitz condition, are assumed for the allowable time delays. Moreover, we focus on the case where the reference is unknown beforehand, which renders the standard prescribed performance control designs under output constraints infeasible. To conquer these challenges, a novel robust prescribed performance control approach is put forward in this paper.Herein, a reverse tuning function is skillfully constructed and automatically generates a performance envelop for the tracking error. In addition, a unified performance analysis framework based on proof by contradiction and the barrier function is established to reveal the inherent robustness of the control system against the time delays. It turns out that the system output tracks the reference with a preassigned settling time and good accuracy,without constraint violations. A comparative simulation on a two-stage chemical reactor is carried out to illustrate the above theoretical findings.

基于Real-time PCR法检测乳粉中牛源性成分定量研究

基于Real-timePCR建立了乳粉中牛源性成分相对定量检测方法,并对牛的特异性引物与探针进行了特异性、灵敏度和稳定性测试。通过模拟不同浓度牛乳粉与马乳粉混合样本,根据其△Ct值的函数关系进行线性拟合进而绘制标准曲线,建立乳粉中牛源性成分的相对定量检测。结果显示,该方法的最低检测限为0.00001 mg/mL,回收率为91.11%~119.2%,组间变异系数≤0.58%、组内变异系数≤1.44%。说明该方法在特异性与稳定性上适用于乳粉中牛源性成分及含量的掺假检测。

一种基于real-time PCR技术的TTV检测方法的建立及应用

目的:本研究旨在开发一种具有更高灵敏度和特异性的TTV检测技术,为揭示TTV在多种疾病过程中的作用提供重要的技术支持。方法:为了更精确、灵敏的检测TTV,本研究分析了目前公布的所有亚型的TTV基因序列,在此基础上建立了一种基于UTR区域的real-time PCR检测方法,并与文献报道应用较为广泛的PCR检测方法进行了对比。结果:本研究建立的方法在1×10^(7)~1×10^(1) copies/μL标准品浓度范围内具有良好的线性关系,相关系数为1.000,斜率为-3.446,检测下限为1×10^(1) copies/μL。重复性试验结果显示,组内变异系数为7.22%,表明本方法重复性、稳定性较强。针对30份临床样本,使用本研究建立的real-time PCR检测方法及目前被多个研究所使用的4套引物进行对比。结果表明,本研究所建立的方法灵敏度显著高于文献中报道的4种方法(P<0.01);Sanger测序结果表明,本方法检测出的30份阳性样本均为TTV,检测特异性为100%。结论:本研究采用基于TaqMan探针的real-time PCR检测方法,检测灵敏性高、覆盖基因型范围广,尤其对于TTV病毒载量较低的情况下能够进行定量检测,对于TTV病毒的致病性及作为免疫标志物的应用提供重要的技术支持。

A Comparative Study between Landmark Based and Real Time Ultrasound Guided Sub Arachnoid Block

Background: Sub arachnoid block (SAB) performed by traditional landmark palpation technique can be inaccurate. This problem is exacerbated by altered patient anatomy due to obesity and age-related changes. A pre-procedural ultrasound scan of the lumbar spine has been shown to be of benefit in guiding lumbar epidural insertion in obstetric patients. Information on the use of real-time ultrasound (RUS) guided SAB, to date, been limited. This study compared RUS guided SAB to traditional landmark guided technique in patients undergoing spinal anesthesia for different surgical procedures. Methods: This was a prospective, single center, comparative observational study conducted in the department of anesthesiology at our center. 560 patients who underwent spinal anesthesia either by landmark based technique or real-time ultrasound-guided methods. The primary outcome was the first attempt success rate of dural puncture when employing the two methods. Results: Baseline characteristics were similar in the two study groups. The first attempt success rate of dural puncture in landmark guided group was 64.3% compared to 72.6% in the ultrasound guided group. This difference was not statistically significant. The procedure performance time was significantly shorter with landmark palpation compared to use of real-time ultrasound guided method. Conclusion: Use of RUS-guided technique does not significantly improve the first attempt success rate of SAB dural puncture during spinal anesthesia compared to the traditional landmark-guided technique.

A real-time performance improvement method for composite time scale

The composite time scale(CTS)provides a stable,accurate,and reliable time scale for modern society.The improvement of CTS’s real-time performance will improve its stability,which strengths related applications’performance.Aiming at this goal,a method achieved by determining the optimal calculation interval and accelerating adjustment stage is proposed in this paper.The determinants of the CTS’s calculation interval(characteristics of the clock ensemble,the measurement noise,the time and frequency synchronization system’s noise and the auxiliary output generator noise floor)are studied and the optimal calculation interval is obtained.We also investigate the effect of ensemble algorithm’s initial parameters on the CTS’s adjustment stage.A strategy to get the reasonable initial parameters of ensemble algorithm is designed.The results show that the adjustment stage can be finished rapidly or even can be shorten to zero with reasonable initial parameters.On this basis,we experimentally generate a distributed CTS with a calculation interval of 500 s and its stability outperforms those of the member clocks when the averaging time is longer than1700 s.The experimental result proves that the CTS’s real-time performance is significantly improved.

Bifurcation and Stability Analysis of Time-Delayed Wheelset System under White Noise Excitation

Considering the impact of time delay in the lateral stiffness of the primary suspension and stochastic disturbances of equivalent conicity on the wheelset system, a stochastic time-delayed wheelset system is established. The wheelset system is transformed into a onedimensional Ito stochastic differential equation using central manifold and stochastic averaging methods. The analysis of the system's stochastic stability is conducted through the maximum Lyapunov exponent and singular boundary theory. The combination of the stationary probability density method and numerical simulation is employed to discuss the types and conditions of stochastic P-bifurcation in the wheelset system. The results indicate that changes in speed and time delay induce stochastic P-bifurcations in the wheelset system, while changes in noise intensity do not lead to stochastic P-bifurcations. Both time delay and equivalent conicity affect the critical speed of the wheelset system, and the critical speed gradually increases with the decrease of time delay and equivalent conicity.

A step to the decentralized real-time timekeeping network

The composite time scale(CTS) provides an accurate and stable time-frequency reference for modern science and technology. Conventional CTS always features a centralized network topology, which means that the CTS is accompanied by a local master clock. This largely restricts the stability and reliability of the CTS. We simulate the restriction and analyze the influence of the master clock on the CTS. It proves that the CTS's long-term stability is also positively related to that of the master clock, until the region dominated by the frequency drift of the H-maser(averaging time longer than ~10~5s).Aiming at this restriction, a real-time clock network is utilized. Based on the network, a real-time CTS referenced by a stable remote master clock is achieved. The experiment comparing two real-time CTSs referenced by a local and a remote master clock respectively reveals that under open-loop steering, the stability of the CTS is improved by referencing to a remote and more stable master clock instead of a local and less stable master clock. In this way, with the help of the proposed scheme, the CTS can be referenced to the most stable master clock within the network in real time, no matter whether it is local or remote, making democratic polycentric timekeeping possible.

The time and frequency system of the Tianma 65 m radio telescope

A time and frequency system is a critical component of Very Long Baseline Interferometry(VLBI)stations,providing stable and reliable standards that directly impact data processing quality.At the Tianma 65 m radio telescope(TMRT),this system has been meticulously designed to ensure long-term reliability and high performance.It incorporates high-performance hydrogen atomic clocks,high-precision time standards,automatic signal switching,and robust system software.This comprehensive approach has enabled the system to achieve long-term reliable operation,successfully supporting both major national engineering tasks and daily scientific observations.The effectiveness of the system is evidenced by its consistent delivery of the precision and stability required for radio astronomy.This article provides an in-depth exploration of the design and operation of the time and frequency system at the Tianma 65 m telescope,examining various aspects of its architecture,implementation,and performance.By sharing these insights,we aim to contribute knowledge that could benefit similar systems at other VLBI stations,greatly advancing radio astronomy infrastructure.

Optimal Multi-Timescale Scheduling of Integrated Energy Systems with Hybrid Energy Storage System Based on Lyapunov Optimization

The economic operation of integrated energy system(IES)faces new challenges such as multi-timescale characteristics of heterogeneous energy sources,and cooperative operation of hybrid energy storage system(HESS).To this end,this paper investigates the multi-timescale rolling opti-mization problem for IES integrated with HESS.Firstly,the architecture of IES with HESS is established,a comparative analysis is conducted to evaluate the advantages of the HESS over a single energy storage system(SESS)in stabilizing power fluctuations.Secondly,the dayahead and real-time scheduling cost functions of IES are established,the day-ahead scheduling mainly depends on operation costs of the components in IES,the real-time optimal scheduling adopts the Lya-punov optimization method to schedule the battery and hydrogen energy storage in each time slot,so as to minimize the real-time average scheduling operation cost,and the problem of day-ahead and real-time scheduling error,which caused by the uncertainty of the energy storage is solved by online optimization.Finally,the proposed model is verified to reduce the scheduling operation cost and the dispatching error by performing an arithmetic example analysis of the IES in Shanghai,which provides a reference for the safe and stable operation of the IES.

Dynamic Event-Triggered Consensus Control for Input Constrained Multi-Agent Systems With a Designable Minimum Inter-Event Time

This paper investigates the consensus control of multi-agent systems(MASs) with constrained input using the dynamic event-triggered mechanism(ETM).Consider the MASs with small-scale networks where a centralized dynamic ETM with global information of the MASs is first designed.Then,a distributed dynamic ETM which only uses local information is developed for the MASs with large-scale networks.It is shown that the semi-global consensus of the MASs can be achieved by the designed bounded control protocol where the Zeno phenomenon is eliminated by a designable minimum inter-event time.In addition,it is easier to find a trade-off between the convergence rate and the minimum inter-event time by an adjustable parameter.Furthermore,the results are extended to regional consensus of the MASs with the bounded control protocol.Numerical simulations show the effectiveness of the proposed approach.

A Novel Method for Linear Systems of Fractional Ordinary Differential Equations with Applications to Time-Fractional PDEs

This paper presents an efficient numerical technique for solving multi-term linear systems of fractional ordinary differential equations(FODEs)which have been widely used in modeling various phenomena in engineering and science.An approximate solution of the system is sought in the formof the finite series over the Müntz polynomials.By using the collocation procedure in the time interval,one gets the linear algebraic system for the coefficient of the expansion which can be easily solved numerically by a standard procedure.This technique also serves as the basis for solving the time-fractional partial differential equations(PDEs).The modified radial basis functions are used for spatial approximation of the solution.The collocation in the solution domain transforms the equation into a system of fractional ordinary differential equations similar to the one mentioned above.Several examples have verified the performance of the proposed novel technique with high accuracy and efficiency.

Research on the Damping Mechanism of Time-Delay Coupled Negative Stiffness Dynamic Absorber in Nonlinear Vibration Damping System

A study was conducted on the effect of time delay and structural parameters on the vibration reduction of a time delayed coupled negative stiffness dynamic absorber in nonlinear vibration reduction systems. Taking dynamic absorbers with different structural and control parameters as examples, the effects of third-order nonlinear coefficients, time-delay control parameters, and negative stiffness coefficients on reducing the replication of the main system were discussed. The nonlinear dynamic absorber has a very good vibration reduction effect at the resonance point of the main system and a nearby area, and when 1 increases to a certain level, the stable region of the system continues to increase. The amplitude curve of the main system of a nonlinear dynamic absorber will generate Hop bifurcation and saddle node bifurcation in the region far from the resonance point, resulting in almost periodic motion and jumping phenomena in the system. For nonlinear dynamic absorbers with determined structural parameters, time-delay feedback control can be adopted to control the amplitude of the main system. For different negative stiffness coefficients, there exists a minimum damping point for the amplitude of the main system under the determined system structural parameters and time-delay feedback control parameters.

Effects of data smoothing and recurrent neural network(RNN)algorithms for real-time forecasting of tunnel boring machine(TBM)performance

Tunnel boring machines(TBMs)have been widely utilised in tunnel construction due to their high efficiency and reliability.Accurately predicting TBM performance can improve project time management,cost control,and risk management.This study aims to use deep learning to develop real-time models for predicting the penetration rate(PR).The models are built using data from the Changsha metro project,and their performances are evaluated using unseen data from the Zhengzhou Metro project.In one-step forecast,the predicted penetration rate follows the trend of the measured penetration rate in both training and testing.The autoregressive integrated moving average(ARIMA)model is compared with the recurrent neural network(RNN)model.The results show that univariate models,which only consider historical penetration rate itself,perform better than multivariate models that take into account multiple geological and operational parameters(GEO and OP).Next,an RNN variant combining time series of penetration rate with the last-step geological and operational parameters is developed,and it performs better than other models.A sensitivity analysis shows that the penetration rate is the most important parameter,while other parameters have a smaller impact on time series forecasting.It is also found that smoothed data are easier to predict with high accuracy.Nevertheless,over-simplified data can lose real characteristics in time series.In conclusion,the RNN variant can accurately predict the next-step penetration rate,and data smoothing is crucial in time series forecasting.This study provides practical guidance for TBM performance forecasting in practical engineering.

The Influence of the Implementation of Trade Facilitation Systems on the Time Required for Creating Delivery Orders: A Case Study of Tema Port

This study evaluated the impact of Ghana’s Integrated Customs Management System (ICUMS), implemented within the National Single Window initiative, on the efficiency of issuing Delivery Orders (DO) at Tema Port. Filling a gap in the existing literature, the research employed a quantitative approach to assess a specific time-related aspect of the cargo clearance process. Employing an Independent t-test on a dataset spanning 2026 Delivery Orders (924 pre-ICUMS and 1102 post-ICUMS) from July 2020 to July 2023, the study investigated ICUMS’s effectiveness in reducing DO issuance time. Results indicate a noteworthy decrease in average DO issuance time, from 11 days pre-implementation to approximately 9 days post-implementation, a reduction validated by statistical analysis through the independent t-test. In light of these findings, the study recommends ongoing refinement of the implementation, reinforcement of trade facilitation measures, and the adoption of best practices from successful global ports. Continuous stakeholder training and regular assessments of ICUMS performance are also endorsed. The study’s implications support the theoretical framework for Single Window systems and carry significant policy implications, emphasizing the need for collaborative efforts to streamline trade facilitation processes driven by Information Technology. Practically, the results serve as a management tool for stakeholders, highlighting areas for targeted interventions to reduce DO issuance times. Methodologically, this research contributes by applying robust statistical analysis to a specific component within the Time Release Study framework, offering a nuanced understanding of trade facilitation systems’ effectiveness in improving cargo clearance processes.

Propagation Dynamics of Forced Pulsating Waves for a Time Periodic Lotka-Volterra Cooperative System with Nonlocal Effects in Shifting Habitats

In this paper, we will concern the existence, asymptotic behaviors and stability of forced pulsating waves for a Lotka-Volterra cooperative system with nonlocal effects under shifting habitats. By using the alternatively-coupling upper-lower solution method, we establish the existence of forced pulsating waves, as long as the shifting speed falls in a finite interval where the endpoints are obtained from KPP-Fisher speeds. The asymptotic behaviors of the forced pulsating waves are derived. Finally, with proper initial, the stability of the forced pulsating waves is studied by the squeezing technique based on the comparison principle.

Prevalence and Antibiotic Resistance of Urinary Tract Pathogens, with Molecular Identification of Klebsiella pneumoniae, Klebsiella oxytoca, and Acinetobacter spp., Using Multiplex Real-Time PCR

Urinary tract infections (UTIs) caused by uropathogens are a significant public health problem, and their treatment primarily relies on antibiotic therapy. However, the increasing global development of antibiotic resistance necessitates updating diagnostic techniques to ensure higher sensitivity and specificity, especially with advancements in science and medicine. This study aimed to evaluate the prevalence of UTIs and antibiotic resistance profiles through urine culture, as well as to identify Klebsiella pneumoniae, Klebsiella oxytoca, and Acinetobacter spp. in urine samples using a molecular approach with multiplex real-time PCR. From May 3 to July 25, 2023, at the Pietro Annigoni Biomolecular Research Center (CERBA) and Saint Camille Hospital of Ouagadougou (HOSCO), 209 urine samples collected from patients with suspected UTIs were analyzed using both urine culture and multiplex real-time PCR. Among the 209 patients, 52.15% were male and 47.85% female, with an average age of 46.87 ± 21.33 years. Urine cultures revealed an overall UTI prevalence of 23.44%, with a prevalence of 8.13% in men versus 15.31% in women (P = 0.023). The bacterial prevalence rates were as follows: Escherichia coli (12.92%), Klebsiella spp. (7.18%), Enterobacter cloacae (1.44%), Staphylococcus aureus (0.96%), and other bacteria. Klebsiella spp. demonstrated 100% resistance to Amoxicillin and Amoxicillin/Clavulanic Acid, while Escherichia coli showed 96.2% and 65.4% resistance to Amoxicillin and Amoxicillin/Clavulanic Acid, respectively. PCR analysis of the target bacteria revealed mono-infection prevalence rates of Klebsiella pneumoniae (10.39%), Klebsiella oxytoca (7.79%), and Acinetobacter spp. (7.79%), along with a co-infection prevalence rate of Klebsiella pneumoniae/Acinetobacter spp. (1.30%). This study demonstrated that PCR, with its high sensitivity and specificity, could effectively distinguish Klebsiella pneumoniae from Klebsiella oxytoca and detect Acinetobacter spp. in less than 24 hours—something urine culture alone could not achieve. The relative ease of automating urine PCR testing, combined with its diagnostic accuracy and rapid turnaround time, makes it a valuable addition to modern medical practice for the laboratory diagnosis of UTIs.