赤红球菌组氨酸激酶的融合表达和功能分析

对赤红球菌的组氨酸激酶基因进行密码子优化，将优化后的组氨酸激酶基因（rhks）构建重组表达质粒pGEX-4T-2-rhks。将此质粒导入到大肠杆菌BL21（DE3）中进行异源表达。在25 ℃和1 mmol/L异丙基-β-D-硫代吡喃半乳糖苷诱导条件下，组氨酸激酶融合蛋白（GST-RHK）获得成功表达，并具有催化活性。经谷胱甘肽琼脂糖亲和层析纯化，获得电泳纯的GST-RHK，其中纯化倍数为3.1，得率为19.5%。该蛋白大小约为72.75 kDa，Km、Vmax和Kcat值分别为20.92 μmol/L、0.17 μmol/（L·min）和1.4 min－1。野生型赤红球菌、组氨酸激酶基因增强株sdrhkE和组氨酸激酶基因敲减株sdrhkD在分别含有苯酚、甲苯、氯苯、异辛烷4 种有机溶剂的培养基中培养，菌株sdrhkD的生长情况都优于野生型赤红球菌，菌株sdrhkE的生长情况都低于野生型赤红球菌。本研究为进一步揭示赤红球菌SD3中组氨酸激酶涉及的信号转导途径与赤红球菌有机溶剂耐受性的关联机制提供一定参考依据。     

Salivary Proteome Changes in Response to Acute Psychological Stress Due to an Oral Exam Simulation in University Students: Effect of an Olfactory Stimulus

The autonomic nervous system (ANS) plays a crucial role both in acute and chronic psychological stress eliciting changes in many local and systemic physiological and biochemical processes. Salivary secretion is also regulated by ANS. In this study, we explored salivary proteome changes produced in thirty-eight University students by a test stress, which simulated an oral exam. Students underwent a relaxation phase followed by the stress test during which an electrocardiogram was recorded. To evaluate the effect of an olfactory stimulus, half of the students were exposed to a pleasant odor diffused in the room throughout the whole session. Saliva samples were collected after the relaxation phase (T0) and the stress test (T1). State anxiety was also evaluated at T0 and T1. Salivary proteins were separated by two-dimensional electrophoresis, and patterns at different times were compared. Spots differentially expressed were trypsin digested and identified by mass spectrometry. Western blot analysis was used to validate proteomic results. Anxiety scores and heart rate changes indicated that the fake exam induced anxiety. Significant changes of α-amylase, polymeric immunoglobulin receptor (PIGR), and immunoglobulin α chain (IGHA) secretion were observed after the stress test was performed in the two conditions. Moreover, the presence of pleasant odor reduced the acute social stress affecting salivary proteome changes. Therefore, saliva proteomic analysis was a useful approach to evaluate the rapid responses associated to an acute stress test also highlighting known biomarkers.     

Pressure control of an electro-hydraulic actuated clutch via novel hysteresis model

This paper mainly focuses on the development of pressure tracking control logic of electro-hydraulic actuators for vehicle application. This is done to improve and ensure the performance of a precise lower-level controller for evolving modern shift control logic. The required performance is obtained by hysteresis model-based feed-forward control and additional feedback control. The hysteresis and the time delay, which adversely affect pressure control, are well known nonlinear behaviors in electro-hydraulic actuators. In order to cope with the hysteresis, a novel hysteresis model is proposed based on a physical phenomenon. A mathematical model based on a characteristic curve obtained in preliminary experiments is presented using only one tuning parameter, and this model can be inverted easily to construct a feed-forward controller. In addition, a feedback controller is designed considering the stability margin of a time delay system. The feedback control inputs ensure compensation of the feed-forward errors caused by model error and uncertainty. The proposed controller is designed to lower computational cost considering applicability for production vehicles. As a result, the developed pressure controller is applied to a transmission control unit of a production vehicle and verified experimentally for various driving scenarios.     

一种5 重real-time PCR筛查转基因水稻方法的建立

以转基因水稻中最常用的CaMV35S启动子、NOS终止子、Cry1Ab/Ac基因、HPT基因及SPS水稻内标基因为研究对象，利用5 种不同的荧光信号（FAM、HEX、Taxas Red、Cy5、Cy5.5）进行多重实时聚合酶链式反应（real-time polymerase chain reaction，real-time PCR）检测方法的研究。通过引物组合筛选、反应体系优化、特异性测试、灵敏度测试、适用性测试等一系列实验，建立了5 重real-time PCR方法，灵敏度可达0.032%。此方法具有灵敏度高、结果准确、通量大等优点，可实现水稻中转基因成分的快速、高效检测。     

Semirigid inclusions in elastic bodies: Mechanical interplay and optimal control

The paper concerns an analysis of an equilibrium problem for 2D elastic body with two semirigid inclusions. It is assumed that inclusions have a joint point, and we investigate a junction problem for these inclusions. The existence of solutions is proved, and different equivalent formulations of the problem are proposed. We investigate a convergence to infinity of a rigidity parameter of the semirigid inclusion. It is proved that in the limit, we obtain an equilibrium problem for the elastic body with a rigid inclusion and a semirigid one. A parameter identification problem is investigated. In particular, the existence of a solution to a suitable optimal control problem is proved.     

On one boundary problem for nonlinear stationary controlled system

The object of study is nonlinear stationary controlled system of ordinary differential equations with constant disturbance in the right part. The problem of constructing the synthesising control function providing the transfer of this system from the initial state to the origin is considered. The sufficiently simple for numerical implementation algorithm of solution of the above-mentioned problem is obtained. It is shown that for local null controllability of the considered system, it is sufficient that the conditions of the Kalman's type were satisfied. In addition, the estimates restricting the choice of initial conditions and external disturbances under which the transfer is guaranteed are obtained. The main idea of the method of construction of the desired control function consists in reducing the original problem to stabilisation of a special kind linear non-stationary system and solving the Cauchy problem for an auxiliary system of ordinary differential equations closed by stabilising control. The simplicity of the realisation of this algorithm is determined by the construction of the auxiliary system and its stabilisation that could be obtained by analytical methods. The effectiveness of the method is illustrated by solving the problem of crane control and its numerical simulation.     

结合优化支持向量机与K-means++的工控系统入侵检测方法

针对工业控制系统传统单一检测算法模型对不同攻击类型检测率和检测速度不佳的问题，提出一种优化支持向量机和K-means++算法结合的入侵检测模型。首先利用主成分分析法（PCA）对原始数据集进行预处理，消除其相关性；其次在粒子群优化（PSO）算法的基础上加入自适应变异过程避免在训练的过程中陷入局部最优解；然后利用自适应变异粒子群优化（AMPSO）算法优化支持向量机的核函数和惩罚参数；最后利用密度中心法改进K-means算法与优化后的支持向量机组合成入侵检测模型，从而实现工业控制系统的异常检测。实验结果表明，所提方法在检测速度和对各类攻击的检测率上得到明显提升。     

Structure and phase-composition of Ti-doped gas atomized Raney-type Ni catalyst precursor alloys

Raney-type Ni precursor alloys containing 75 at.% Al and doped with 0, 0.75, 1.5 and 3.0 at.% Ti have been produced by a gas atomization process. The resulting powders have been classified by size fraction with subsequent investigation by powder XRD, SEM and EDX analysis. The undoped powders contain, as expected, the phases Ni₂Al₃, NiAl₃ and an Al-eutectic. The Ti-doped powders contain an additional phase with the TiAl₃ DO₂₂ crystal structure. However, quantitative analysis of the XRD results indicate a far greater fraction of the TiAl₃ phase is present than could be accounted for by a simple mass balance on Ti. This appears to be a (Ti_xNi_1−x)Al₃ phase in which higher cooling rates favour small x (low Ti-site occupancy by Ti atoms). SEM and EDX analysis reveal that virtually all the available Ti is contained within the TiAl₃ phase, with negligible Ti dissolved in either the Ni₂Al₃ or NiAl₃ phases.     

An energy-saving nonlinear position control strategy for electro-hydraulic servo systems

The electro-hydraulic servo system (EHSS) demonstrates numerous advantages in size and performance compared to other actuation methods. Oftentimes, its utilization in industrial and machinery settings is limited by its inferior efficiency. In this paper, a nonlinear backstepping control algorithm with an energy-saving approach is proposed for position control in the EHSS. To achieve improved efficiency, two control valves including a proportional directional valve (PDV) and a proportional relief valve (PRV) are used to achieve the control objectives. To design the control algorithm, the state space model equations of the system are transformed to their normal form and the control law through the PDV is designed using a backstepping approach for position tracking. Then, another nonlinear set of laws is derived to achieve energy-saving through the PRV input. This control design method, based on the normal form representation, imposes internal dynamics on the closed-loop system. The stability of the internal dynamics is analyzed in special cases of operation. Experimental results verify that both tracking and energy-saving objectives are satisfied for the closed-loop system.