Interaction between Thymidylate Synthase and Its Cognate mRNA in Zebrafish Embryos

Thymidylate synthase (TS), which catalyzes the de novo synthesis of dUMP, is an important target for cancer therapy. In this report, the effects of 5-fluorouracil (5-FU) and ZD1694 on the regulation of TS gene expression were evaluated in zebrafish embryos. Our results revealed that the expression of TS was increased by about six-fold when embryos were treated with 1.0 mu M 5-FU and there was a greater than 10-fold increase in the TS protein level after treatment with 0.4 mu M ZD1694. Northern blot analysis confirmed that expression of TS mRNA was identical in treated or untreated embryos. Gel shift and immunoprecipitation assays revealed that zebrafish TS was specifically bound with its cognate mRNA in vitro and in vivo. We identified a 20 nt RNA sequence, TS:N20, localized to the 5'-UTR of TS mRNA, which corresponded to nt 13-32; TS:N20 bound to the TS protein with an affinity similar to that of the full-length TS mRNA. The MFold program predicted that TS:N20 formed a stable stem-loop structure similar to that of the cis-acting element found in human TS mRNA. Variant RNAs with either a deletion or mutation in the core motif of TS:N20 were unable to bind to the TS protein. In vitro translation experiments, using the rabbit lysate system, confirmed that zebrafish TS mRNA translation was significantly repressed when an excess amount of TS protein was included in the system. Additionally, a TS stability experiment confirmed that treatment of zebrafish embryos with 5-FU could increase the TS stability significantly, and the half life of TS protein was about 2.7 times longer than in untreated embryos. Our study revealed a structural requirement for the interaction of TS RNA with TS protein. These findings also demonstrated that the increase in TS protein induced by 5-FU occurs at the post-transcriptional level and that increased stability and translation efficiency both contributed to the increase in TS protein levels induced by TS inhibitors.

估计从UUV航行参数的混合坐标系下的EKF算法

在主从式UUV 协作系统中，由于定位和导航的需要，要求尽快估计出从UUV 的航行参数，但通常所用的递推最小二乘（RLS）算法，其初始方位测量对滤波结果影响大且存在收敛速度慢、计算精度低的缺点，难以满足应用需求，而推广卡尔曼滤波（EKF）算法能较好地克服上述问题。在直角坐标系下（CEKF），方位信息与距离信息相互耦合导致初始振荡剧烈，改为混合坐标系（MEKF）后问题得到了极大的改善。最后，通过仿真及现场试验验证了此改进方法的有效性。

基于局域网的多水下机器人仿真系统设计与实现

多水下机器人仿真系统是一个能够对多水下机器人系统的体系结构、协调控制、路径规划、学习算法等进行演示验证的分布式实时数字仿真系统,是开展多水下机器人技术研究的基础和有效手段.讨论了应用基于局域网的分布式仿真技术来解决多水下机器人系统仿真的问题,并详细说明了仿真系统的硬件组成和软件总体设计.

基于DSRC的多功能不停车收费系统

DSRC标准是ISO/TC204制定的智能交通系统(ITS)中车—路信息通讯的协议。采用双片式ETC电子标签结合双界面IC卡的储值卡方案形成的多功能不停车收费系统,充分兼容了当前国内普遍应用的IC卡半自动收费方式,集中了IC卡收费系统和ETC收费系统的优点,方便、快捷、通行能力强,并解决了收费口的交通拥挤现象。

地震力对高架桥梁上轨道车辆的动态分析与安全性研究

The stability and derailment behavior analysis of railway vehicle system has been discussed by many papers in the past. In stability, give first place to consider hunting behavior of vehicle, therefore most of papers was only consider lateral and yaw motion, but vertical motion is the important factor in derailment behavior, and it will be quite effect in stability. We will probe the running stability and derailment behavior of railway vehicle moving on the viaduct in this paper. In this paper, we use Nadal’s formula to get the derailment quotient. In this paper, the railway vehicle is considered to be three subsystems, carbody, bogie and wheelset. There are secondary suspension systems between carbody and bogies, and primary suspension systems connecting bogies and wheelsets. A vehicle with vertical, lateral, roll, and yaw directions motion is considered to derive the mathematical equations. A vehicle with three-dimensional model has 16 degrees of freedom is used to develop the equations of train motion. In this study, results show that the track shift force and derailment factor increase with an increase of ground acceleration. But for the track shift force and derailment factor, the effects of track irregularities and train speed are very small. Key words: earthquake, railway vehicle, viaduct, derailment factor.

A homogeneous DNA hybridization system by using a new luminescence terbium chelate

Homogeneous DNA hybridization assay based on the luminescence resonance energy transfer (LRET) from a new luminescence terbium chelate, N,N,N-1,N-1-[2,6-bis(3'-aminomethyl-1'-pyrazolyl)-4-phenylpyridine]tetrakis(acetic acid) (BPTA)-Tb3+ (lambda(ex) = 325 nm and lambda(em) = 545 nm) to an organic dye, Cy3 (A,. = 548 nm and A,. = 565 nm), has been developed. In the system, two DNA probes whose sequences are complementary to the two different consecutive sequences of a target DNA are used; one of the probes is labeled with the Tb3+ chelate at the T-end, and the other is with Cy3 at the 5'-end. Labeling of the Tb3+ chelate is accomplished via the linkage of a biotin-labeled DNA probe with the Tb3+ chelate-labeled streptavidin. Strong sensitized emission of Cy3 was observed upon excitation of the Tb3+ chelate at 325 run, when the two probe DNAs were hybridized with the target DNA. The sensitivity of the assay was very high compared with those of the previous homogeneous-format assays using the conventional organic dyes; the detection limit of the present assay is about 30 pM of the target DNA strand.