利用反义RNA技术进行木质素生物合成调控的研究

木质素是一类酚类次生代谢产物，在植物体内行使重要的生理功能，但它却是形成造纸污染的主要来源。利用基因工程手段，在分子水平调节木质素的生物合成，降低木质素的含量或改变组分以培育适合造纸的植物原料树种具有较大的应用价值和环保效益。本研究利用反义RNA技术，主要围绕木质素合成三种相关酶咖啡酸-O-甲基转移酶（COMT）、咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)、4-香豆酸：辅酶A连接酶（4CL）的基因对植物木质素生物合成途径调节的研究，取得如下进展： 1.农杆菌介导法将COMT和CCoAOMT基因的单价和双价的反义表达载体导入烟草，比较了两个甲基化酶的功能。PCR-Southern和Northern点杂交结果表明反义基因已整合到烟草基因组DNA上，并在转录水平表达。两种反义基因对木质素生物合成调节的效果显示，CCoAOMT能更有效地调节木质素生物总量的合成，COMT仅特异调节S木质素的合成。表达反义CCoAOMT基因的转基因毛白杨，内源CCoAOMT基因的表达在转录和蛋白水平均受到抑制，最终引起转基因植株木质素含量普遍降低，最多降低达26.20%，筛选出木质素含量下降10%以上的转基因毛白杨株系8个，为源头治理造纸废水污染奠定了基础。 2． 对克隆的4CL基因进行了表达特性分析， RT-PCR分析表明，分离的毛白杨4CL基因主要在木质部丰富表达，叶中表达量较少，树皮中不表达。在毛白杨的一个生长季，该基因表达显示明显的双锋特征，该表达模式与木材早材和晚材的发育时期相吻合，表明分离的毛白杨4CL基因与木质素的生物合成密切相关。农杆菌介导法将反义4CL基因导入烟草和毛白杨，利用分子生物学检测手段对转化植株进行筛选，获得批量转基因植株。Klason木质素含量测定分析表明，抑制内源4CL基因表达，能有效降低转基因植物中的木质素含量，且不影响植株正常生长和发育以及碳水化合物的合成。转基因毛白杨的茎杆上一些区域呈红棕色，颜色的深度与转基因毛白杨木质素含量的下降幅度呈一定的正相关性，颜色变化可作为转基因植株筛选的一个辅助指标。现已获得木质素含量下降10%以上的转基因株系3个，最多下降达41.73%，可供中试与制浆实验，为培育低木质素环保型毛白杨提供理论与实践依据。 3.为了优化现有的表达框架，使目的基因更有效地调节木质素的生物合成，应用PCR技术从毛白杨基因组中分离得到C4H(肉桂酸4—羟基化酶)基因启动子片段（GenBank注册号：AY351673）。GUS荧光活性分析和组织化学染色显示，该启动子在一些木质化的组织和器官中特异表达，随着组织成熟度和木质化程度的增加，表达活性逐渐增强，并且该启动子受伤诱导。反义CCoAOMT基因在C4H启动子的调控下，会引起转基因烟草木质素均有不同程度的减少，但不影响碳向碳水化合物的转换合成，对植物的生长发育也无明显负效应。这些结果证明了从毛白杨中分离的C4H 启动子可以应用于造纸原料树种材性改良的遗传工程操作。 4．首次从水稻中华10号（Oryza sativa L. ssp. japonica）分离了CCoAOMT基因家族的三个成员，对其基因结构及表达特性的分析表明，该基因家族的三个成员与水稻的木质化进程关系密切，研究结果有助于了解单子叶植物中的甲基化途径发生机制，为高产水稻抗倒伏和茎杆饲料作物的遗传改良奠定了基础。

Expression of CD175 (Tn), CD175s (sialosyl-Tn) and CD176 (Thomsen-Friedenreich antigen) on malignant human hematopoietic cells

The expression of the histo-blood group carbohydrate structures T-nouvelle (Tn, CD175), sialylated To (CD175s) and the Thomsen-Friedenreich disaccharide (TF, CD176) on human leukemia cell lines was analyzed by their reactivity with specific monoclonal ant

栅扫描重离子束治癌束流配送系统对运动靶区的适形放疗(Ⅱ)可行性实验

在栅扫描束流配送系统下,进行了重离子束对运动靶体进行适形照射的可行性实验研究．利用实时修正束流扫描参数的方法,使得束流追踪靶体在横向上的运动；在纵向上利用一个机械驱动的束流降能装置(称深度扫描器)迅速调节束流能量,使得重离子束高剂量的Bragg峰区落在运动靶体需治疗的断层之上．实验结果表明：栅扫描器主动补偿靶体横向运动及深度扫描器补偿靶体纵向运动是可行的．

栅扫描重离子束治癌束流配送系统对运动靶区的适形放疗冰(Ⅰ)模拟计算

从理论上研究了在主动型栅扫描束流配送系统下,靶体运动对靶区剂量均匀性以及重离子治疗适形程度的影响.模拟计算了在实验测量得到的脉冲束流照射下,栅扫描系统引导束流对运动靶体实施照射后,靶体运动模式与靶区各等能量断层剂量分布均匀性的关系.由这些模拟计算结果,得到了一些针对运动靶体提高靶区剂量分布均匀性的策略.模拟计算提供了一种评估运动靶体上剂量分布的有效手段,其结果及内涵为今后该方向上的可行性实验研究奠定了坚实的基础.

Conformal irradiation to moving targets in heavy ion radiotherapy with raster-scanning beam delivery system: (II) feasibility experiments

Within the framework of the pilot heavy-ion therapy facility at GSI equipped with an active beam delivery system of advanced raster scanning technique, a feasibility study on actively conformal heavy-ion irradiation to moving tumors has been experimentally conducted. Laterally, real-time corrections to the beam scanning parameters by the raster scanner, leading to an active beam tracing, compensate for the lateral motion of a target volume. Longitudinally, a mechanically driven wedge energy degrader (called depth scanner) is applied to adjust the beam energy so as to locate the high-dose Bragg peak of heavy ion beam to the slice under treatment for the moving target volume. It has been experimentally shown that compensations for lateral target motion by the raster scanner and longitudinal target shift by the depth scanner are feasible.

Simulations to design an online motion compensation system for scanned particle beams

Respiration-induced target motion is a major problem in intensity-modulated radiation therapy. Beam segments are delivered serially to form the total dose distribution. In the presence of motion, the spatial relation between dose deposition from different segments will be lost. Usually, this results in over-and underdosage. Besides such interplay effects between target motion and dynamic beam delivery as known from photon therapy, changes in internal density have an impact on delivered dose for intensity-modulated charged particle therapy. In this study, we have analysed interplay effects between raster scanned carbon ion beams and target motion. Furthermore, the potential of an online motion strategy was assessed in several simulations. An extended version of the clinical treatment planning software was used to calculate dose distributions to moving targets with and without motion compensation. For motion compensation, each individual ion pencil beam tracked the planned target position in the lateral aswell as longitudinal direction. Target translations and rotations, including changes in internal density, were simulated. Target motion simulating breathing resulted in severe degradation of delivered dose distributions. For example, for motion amplitudes of +/- 15 mm, only 47% of the target volume received 80% of the planned dose. Unpredictability of resulting dose distributions was demonstrated by varying motion parameters. On the other hand, motion compensation allowed for dose distributions for moving targets comparable to those for static targets. Even limited compensation precision (standard deviation similar to 2 mm), introduced to simulate possible limitations of real-time target tracking, resulted in less than 3% loss in dose homogeneity.

Stratigraphic Model Predictions of Geoacoustic Properties

Geoacoustic properties of the seabed have a controlling role in the propagation and reverberation of sound in shallow-water environments. Several techniques are available to quantify the important properties but are usually unable to adequately sample the region of interest. In this paper, we explore the potential for obtaining geotechnical properties from a process-based stratigraphic model. Grain-size predictions from the stratigraphic model are combined with two acoustic models to estimate sound speed with distance across the New Jersey continental shelf and with depth below the seabed. Model predictions are compared to two independent sets of data: 1) Surficial sound speeds obtained through direct measurement using in situ compressional wave probes, and 2) sound speed as a function of depth obtained through inversion of seabed reflection measurements. In water depths less than 100 m, the model predictions produce a trend of decreasing grain-size and sound speed with increasing water depth as similarly observed in the measured surficial data. In water depths between 100 and 130 m, the model predictions exhibit an increase in sound speed that was not observed in the measured surficial data. A closer comparison indicates that the grain-sizes predicted for the surficial sediments are generally too small producing sound speeds that are too slow. The predicted sound speeds also tend to be too slow for sediments 0.5-20 m below the seabed in water depths greater than 100 m. However, in water depths less than 100 m, the sound speeds between 0.5-20-m subbottom depth are generally too fast. There are several reasons for the discrepancies including the stratigraphic model was limited to two dimensions, the model was unable to simulate biologic processes responsible for the high sound-speed shell material common in the model area, and incomplete geological records necessary to accurately predict grain-size