Evaluation of reference genes for quantitative real-time PCR analysis of gene expression during early development processes of the tongue sole(Cynoglossus semilaevis)

Differential expression of genes is crucial to growth and development of fish. To select the appropriate genes for gene normalization during Cynoglossus semilaevis early developmental process, eight candidate reference genes （ACTB, B2M, EF1A, GADPH, RPL7, TUBA, UBCE and 18S） were tested for their adequacy by using quantitative real-time PCR. The results showed that the expression of all the examined genes exhibited tissue dependent variations in the mature C. semilaevis. EFIA was listed as the most stable reference among the 14 tissues by RefFinder. Furthermore, the recommended comprehensive ranking of the stability determined by RefFinder showed that 18S was the most stable gene during the early developmental stages （from oosphere to 90 days old） in this study. However, when divided the Ct value data of the above mentioned early developmental stages into two separate periods （embryo and post-hatching periods）, TUBA and 18S represented the most stable references of these two developmental periods, respectively. Consequently, the reference gene should be carefully and accurately chosen even for studies of the same species at various developmental processes. The relevant data may help in selecting appropriate reference genes for mRNA expression analysis, and is of great value in the studies of fish growth and development.

Selection of Reference Genes for Gene Expression Analysis in Nilaparvata lugens with Different Levels of Virulence on Rice by Quantitative Real-Time PCR

The brown planthopper Nilaparvata lugens Stal （Homoptera： Delphacidae） can cause hopperburn by feeding on rice and also can transmit the grassy stunt disease. Resistant rice varieties have been developed, but several N. lugens strains can recover their virulence to these resistant rice varieties. In the present study, reference genes with stable expression levels in N. lugens populations showed different levels of virulence to susceptible and resistant rice varieties. The expression of six candidate reference genes in N. lugens feeding on susceptible and resistant rice varieties was analyzed. These genes were evaluated for their potential use in the analysis of differential gene expression. Polymerase chain reaction data was generated from N. lugens, including two different treatments （resistant or susceptible rice） and three virulent N. lugens populations. Three software programs （BestKeeper, Normfinder and geNorm） were used to assess the candidate reference genes. Both geNorm and Normfinder identified the genes 18S, E-ACT, E-TUB and a-TUB as the most stable reference genes. BestKeeper identified ETIF1 as the optimal reference gene with the least overall variation, whereas 18S and a-TUB were the second and third most stably expressed genes, respectively. Therefore, we concluded that the genes 18S and a-TUB were the most suitable reference genes in N. lugens. These results will facilitate future transcript profiling studies on N. lugens populations that show variation in virulence levels on different rice varieties.

Reference genes for quantitative real-time PCR analysis and quantitative expression of P5CS in Agropyron mongolicum under drought stress

Reference genes, stably expressing in different tissues and cells, are commonly used as the references in expression analysis. Selecting the optimum reference gene is crucial to the success of experiments. In this study, the expression stabilities of nine common reference genes, including ACT2, 18 S r RNA, APRT, EF-1α, RNA POL II, TUBα, TUBβ, GAPDH and TLF of Agropyron mongolicum, were studied under drought condition. Among them, 18 S r RNA was found to be the most optimum reference gene under drought stress by the analyzing of ge Norm and Norm Finder software. Quantitative expression levels of P5 CS using 18 S r RNA as the reference gene, and proline contents under drought stress in A. mongolicum were further operated, and we found the expression level of P5 CS gene and proline content had a significantly positive relationship（R^2=0.7763, P〈0.05）. This study established and validated 18 S r RNA as the reference genes in A. mongolicum under drought stress, providing a powerful tool for the quantitative expression analysis of drought genes in A. mongolicum.

A Cross-Reference Method for Nonlinear Time Series Analysis in Semi-Blind Case

In this paper, we propose a cross reference method for nonlinear time series analyzing in semi blind case, that is, the dynamic equations modeling the time series are known but the corresponding parameters are not. The tasks of noise reduction and parameter estimation which were fulfilled separately before are combined iteratively. With the positive interaction between the two processing modules, the method is somewhat superior. Some prior work can be viewed as special cases of this general framework. The simulations for noise reduction and parameter estimation of contaminated chaotic time series show improved performance of our method compared with previous work.

Impacts of Reference Time Series on the Homogenization of Radiosonde Temperature

Using radiosonde temperatures of 92 selected stations in China,the uncertainties in homogenization processes caused by different reference series,including nighttime temperature,the NCEP （National Centers for Environmental Prediction） and ERA-40 （European Centre for Medium-Range Weather Forecasts） forecasting background,are examined via a two-phase regression approach.Although the results showed limited consistency in the temporal and spatial distribution of identified break points （BPs） in the context of metadata events of instrument model change and correction method,significant uncertainties still existed in BP identification,adjustment,and impact on the estimated trend.Reanalysis reference series generally led to more BP identification in homogenization.However,those differences were parts of global climatic shifts,which may have confused the BP calculations.Discontinuities also existed in the reanalysis series due to changes in the satellite input.The adjustment values deduced from the reanalysis series ranged widely and were larger than those from the nighttime series and,therefore,impacted the estimated temperature trend.

Early-season crop type mapping using 30-m reference time series

Early-season crop type mapping could provide important information for crop growth monitoring and yield prediction,but the lack of ground-surveyed training samples is the main challenge for crop type identification.Although reference time series based method(RBM)has been proposed to identify crop types without the use of ground-surveyed training samples,the methods are not suitable for study regions with small field size because the reference time series are mainly generated using data set with low spatial resolution.As the combination of Landsat data and Sentinel-2 data could increase the temporal resolution of 30-m image time series,we improved the RBM by generating reference normalized difference vegetation index(NDVI)/enhanced vegetation index(EVI)time series at 30-m resolution(30-m RBM)using both Landsat and Sentinel-2 data,then tried to estimate the potential of the reference NDVI/EVI time series for crop identification at early season.As a test case,we tried to use the 30-m RBM to identify major crop types in Hengshui,China at early season of 2018,the results showed that when the time series of the entire growing season were used for classification,overall classification accuracies of the 30-m RBM were higher than 95%,which were similar to the accuracies acquired using the ground-surveyed training samples.In addition,cotton,spring maize and summer maize distribution could be accurately generated 8,6 and 8 weeks before their harvest using the 30-m RBM;but winter wheat can only be accurately identified around the harvest time phase.Finally,NDVI outperformed EVI for crop type classification as NDVI had better separability for distinguishing crops at the green-up time phases.Comparing with the previous RBM,advantage of 30-m RBM is that the method could use the samples of the small fields to generate reference time series and process image time series with missing value for early-season crop casification;while,samples collected from multiple years should be futher used so that the reference time series could contain more crop growth conditions.

Application of time series modeling to a national reference frame realization

This paper presents an option for modern dynamic terrestrial reference system realization in Uzbekistan for user needs. An additive model is explored to predict patterns of time series and investigate means of constructing forecast time series models in the future. The main components(trend, periodical, and irregular) of the KIUB(DORIS) and KIT3, TASH, MADK, and MTAL(GNSS) international stations coordinate time series were investigated. It was shown that seasonal nonlinear trends occurred both in the height(U) component of all stations and the east(E) component of high mountainous stations such as MTAL and MADK. The seasonal periodical portion of the time series determined from the additive model has a complicated pattern for all sites and can be explained as both hydrological signals in the region and improvement of observational quality. Amplitudes of the best-fitting sinusoids in the North component ranged between 1.73 and 8.76 mm; the East component ranged between 0.82 and 11.92 mm; and the Up component ranged between 3.11 and 40.81 mm. Regression analysis of the irregular portion of the height component of the two techniques at the Kitab station using tropospheric parameters(pressure and temperature) was confirmed as only 57% of the stochastic portion of the time series.

Robustness Analysis of Discrete-time Indirect Model Reference Adaptive Control with Normalized Adaptive Laws

For a class of discrete-time systems with unmodeled dynamics and bounded disturbance, the design and analysis of robust indirect model reference adaptive control （MRAC） with normalized adaptive law are investigated. The main work includes three parts. Firstly, it is shown that the constructed parameter estimation algorithm not only possesses the same properties as those of traditional estimation algorithms, but also avoids the possibility of division by zero. Secondly, by establishing a relationship between the plant parameter estimate and the controller parameter estimate, some similar properties of the latter are also established. Thirdly, by using the relationship between the normalizing signal and all the signals of the closed-loop system, and some important mathematical tools on discrete-time systems, as in the continuous-time case, a systematic stability and robustness analysis approach to the discrete indirect robust MRAC scheme is developed rigorously.

Studies on Time Reference and Pronominal Elements

Agrammatism is a case of a more general linguistic impairment known as Broca's aphasia.For recent studies just qualitatively analyzed the similarities between time reference and pronominal elements and no experiment have been done.In this paper,the author first introduced the previous studies on time reference and pronominal elements,then designed an experiment to quantitatively test the similarities between them from the aspect of discourse linking.

Discrete-time Direct Model Reference Adaptive Control： A Systematic Approach

为直接模型引用的分离时间的系统，设计和分析的一个类，有规范的适应法律的适应控制(MRAC ) 被调查。我们责骂在输入和产量之间的 p 和 2 个关系性质上的分离时间的结论，和分离时间的交换词根 1 和 2。我们也建立分离时间的适应法律的性质，定义使正常化的信号，并且联系有在靠近环的系统的所有信号的信号。因此，分离时间的 MRAC 计划的稳定性和集中性质作为在连续时间的案例中以一种系统的方式严厉地被分析。

The New Method for CNC Trajectory Machining Motion Planning Based on Reference Time

In order to make motion planning fitting practice,many characteristic of CNC trajectory motion are discussed, such as the geometric function,the motion and the time.It is found that the relation between orbit function and motional parame- ter,so the differential equation about the trajectory motion be set-up by the goal of trajectory motion.The actual motion process is defined as reference time to link planning and practice.Present a new movement planning method based on self-defining time.At rest state,the differential simultaneous equation can be calculated according geometric characteristic analysis,it can be get that simple function consisted of coordinate and reference time variants.At motive state,dynamic parameter can be worked out accord- ing practical value of reference time,It is proved by experiment and simulation that it is a good way to control geometry and motion comprehensively,to reduce computation times and to increase the ability of environmental adaptation for path

A Systematic Analysis Approach to Discrete-time Indirect Model Reference Adaptive Control

这份报纸论述间接模型引用的设计和分析有为分离时间的系统的一个类的规范的适应法律的适应控制(MRAC ) 。主要工作包括三部分。第一，构造工厂参数评价算法不仅拥有传统的评价算法的象那些的一样的性质而且在零避免分割的可能性。第二，由发现在工厂参数之间的关系，估计和控制器参数估计并且用工厂参数的性质估计，控制器参数估计的类似的性质也被建立。第三，基于关系，在使正常化之间的性质发信号并且在靠近环的系统并且在分离时间的系统上的一些重要数学工具上的所有信号作为在连续时间的盒子中，到分离间接 MRAC 计划的系统的稳定性和集中分析途径严厉地被开发。

Selection of Reference Genes in Equine White Blood Cells for Real Time PCR Normalization Following Extracorporeal Shock Wave Therapy

Selection of proper reference genes (RGs) is an essential step needed for accurate normalization of results from genomic studies. Expression of RGs is regulated by many factors such as species, age, gender, type of tissue, the presence of disease, and the administration of therapeutic treatment. The aim of the present study was to identify optimal RGs in a set of blood samples collected at different time points (0, 24, 48, 72 h) from horses following administration of extracorporeal shock wave therapy (ESWT). The mRNA expression of twelve RGs: HPRT1, ACTB, HSP90A, SDHA, GUSB, B2M, UBC, NONO, TBP, H6PD, RPL32, GAPDH was determined using real time quantitative polymerase chain reaction (qPCR). An SAS program developed on the algorithm of geNorm, SASqPCR, was used to determine stability of the expression and the number of optimal RGs. The results showed that the range of quantification cycle (Cq) values of the evaluated genes varied between 17 and 26 cycles, and that one optimal RG, ACTB, was sufficient for normalization of gene expression. Results of stability of expression demonstrated that ACTB was the optimal choice for all the samples studied. Notably, in samples collected at 72 h post ESWT, TBP showed a significant change in the expression level, and was not suitable for use as a RG. These results substantiate the importance of validating and selecting an appropriate RG.

Accurate time synchronization of power reference station based on BD3 system

A Beidou 3(BD3)system-based power reference station can provide high-precision time synchronization for power distribution systems by sending synchronization data packets to devices in a multi-hop routing fashion.However,optimizing route selection to reduce both time synchronization error and delay is a challenging problem.In this paper,we establish a software-defined network-enabled power reference station time synchronization framework based on BD3.Then,we formulate the joint problem to minimize cumulative synchronization error and delay through multi-hop route selection optimization.A back propagation(BP)neural network-improved intelligent time synchronization route selection algorithm named BP-RS is proposed to learn the optimal route selection,which uses a BP neural network to dynamically adjust the exploration factor to achieve rapid convergence.Simulation results show the superior performance of BP-RS in synchronization delay,synchronization error,and adaptability with changing routing topologies.

Direct model reference adaptive control using K_p = LDU factorization for multivariable discrete-time plants

For a large class of discrete-time multivariable plants with arbitrary relative degrees, the design and analysis of the direct model reference adaptive control scheme are investigated under less restrictive assumptions. The algorithm is based on a new parametrization derived from the high frequency gain matrix factorization Kp=LDU under the condition that the signs of the leading principal minors of/fp are known. By reproving the discrete-time Lp and L2σ norm relationship between inputs and outputs, establishing the properties of discrete-time adaptive law, defining the normalizing signal, and relating the signal with all signals in the closed-loop system, the stability and convergence of the discrete-time multivariable model reference adaptive control scheme are analyzed rigorously in a systematic fashion as in the continuous-time case.

A Method for Detecting Adhesive Related-Factors of Streptococcus suis Serotype 2 by Real-time PCR

[Objective] This study aimed to establish a method for quantitative detection of mRNA transcriptional level of SS2 adhesive related-factors of Streptococcus suis serotype 2 （SS2） by fluorescent quantitative PCR. []Vlethod] The gene fragments en- coding SS2 adhesive related-factors MRP, FBPS and CPS2J and a housekeeping gene aroA were amplified by reverse transcription PCR from the total RNA of SS2, cloned, and sequenced. The recombinant plasmids containing the target genes were constructed, and used as templates in Real-time PCR. [Result] Dynamic curves, stan- dard curves and melting curves of the adhesive related-factors and aroA were ob- tained by the optimized Real-time PCR system. The standard curves showed a good linear relationship between template copy number and circulation number, and the correlation coefficients （FF） of the standard curves were over 0.995. Also, these as- says were highly specific a^d there was single specific melting peak for every gene. Moreover, the assays were highly sensitive and had a detection limit of 1.0×102 copies in 1 μl of initial templates. Finally, it was highly repeatable and had a coeffi- cient of variation less than 2% for intra-assay. [Conclusion] This study will provide a way to reveal the adhesion mechanism of SS2 to different host cells at molecular level.

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.

A New Perspective on Time and Gravity

This paper presents a hypothesis regarding the existence of time fused in spacetime, assuming that time possesses the properties of both a particle and a field. This duality is referred to as the field-particle of time (FPT). The analysis shows that when the FPT moves through matter, it causes time dilation. The FPT is also a significant element that appears in relativistic kinetic energy (KE = (γ - 1) · mc2). Accelerating matter to near the speed of light requires relativistic energy approaching infinity, which corresponds to the relativistic kinetic energy. Meanwhile, the potential energy (PE = mc2) from the rest mass remains constant. Then, the mass-energy equation can be rearranged in terms of PE and KE, as shown in E = (1 + (γ - 1)) · mc2. The relativistic energy of the FPT also directly affects the gravitational attraction of matter. It transfers energy to each other through spacetime. The analysis demonstrates that the gravitational force is inversely proportional to the distance squared, following Newton’s law of gravity, and it varies with the relative velocity of matter. The relationship equation between relative time and the gravitational constant indicates that a higher intensity of the gravitational field leads to a slower reference time for matter, in accordance with the general theory of relativity. A thought experiment presents a comparison of two atomic clocks placed in different locations. The first one is placed in a room temperature, around 25°C, on the surface of the Earth, and the second one is placed in high-density areas. The analysis, considering the presence of the FPT, shows that the reference time slows down in high-density areas. Therefore, the second clock must be noticeably slower than the first one, indicating the existence of the FPT passing through both atomic clocks at different speeds.

Real-Time Discrete Adaptive Control of Robot Arm Based on Digital Signal Processing

A discrete model reference adaptive controller of robot arm is obtained by integrating the reduced dynamic model of robot, model reference adaptive control （MRAC） and digital signal processing （DSP） computer system into an electromechanical system. With the DSP computer system, the control signal of each joint of the robot arm can be processed in real time and independently. The simulation and experiment results show that with the control strategy, the robot achieved a good trajectory following precision, a good decoupling performance and a high real-time adaptivity.

Approach to inter-satellite time synchronization for micro-satellite cluster

Micro-satellite cluster enables a whole new class of missions for communications, remote sensing, and scientific research for both civilian and military purposes. Synchronizing the time of the satellites in a cluster is important for both cluster sensing capabilities and its autonomous operating. However, the existing time synchronization methods are not suitable for microsatellite cluster, because it requires too many human interventions and occupies too much ground control resource. Although, data post-process may realize the equivalent time synchronization, it requires processing time and powerful computing ability on the ground, which cannot be implemented by cluster itself. In order to autonomously establish and maintain the time benchmark in a cluster, we propose a compact time difference compensation system（TDCS）, which is a kind of time control loop that dynamically adjusts the satellite reference frequency according to the time difference. Consequently, the time synchronization in the cluster can be autonomously achieved on-orbit by synchronizing the clock of other satellites to a chosen one＇s. The experimental result shows that the standard deviation of time synchronization is about 102 ps when the carrier to noise ratio（CNR） is 95 d BHz, and the standard deviation of corresponding frequency difference is approximately0.36 Hz.