HIV-1 C亚型密码子优化gp120负载人DC疫苗的构建及体外功能

目的构建HIV-1C亚型gp120负载人树突状细胞(dentriti ccell,DC)疫苗,并对其体外功能进行初步检测。方法利用Amaxa细胞核转染技术将pcDNA3.1-gp120质粒转染至人成熟DC,以Western blot检测gp120的表达。通过流式细胞仪检测DC表面共刺激分子的变化、混合淋巴细胞反应、CD8+T细胞表面活化分子CD25的表达及其分泌IFN-γ的变化。结果通过Western blot检测,gp120在DC中得到了正确表达。经流式细胞仪检测,DC表面分子CD80表达率由刺激前的33.34%上升至43.20%,CD86表达率由刺激前的60.08%上升至90.34%;负载gp120DC刺激淋巴细胞增殖率为86.72%;CD8+T细胞表面分子CD25表达率由刺激前的5.27%上升至74.21%,IFN-γ的表达率达37%。结论负载了HIV-1gp120的人树突状细胞能够显著刺激淋巴细胞的增殖、增强CD8+T细胞表面活化分子CD25表达以及促进CD8+T细胞分泌IFN-γ,为下一步DC治疗性疫苗的体内研究奠定基础。

HIV-1 C亚型gp120重组腺病毒载体的构建及gp120负载人树突状细胞疫苗的初步研究

获得性免疫缺陷综合征(AIDS)是一种由人类免疫缺陷病毒(HIV)引起的，以全身免疫系统受到严重损害为特征的传染性疾病。从目前HIV-1的流行趋势来看，HIV-1 C亚型已经成为全球最主要的流行株之一，因此，针对HIV-1 C亚型的疫苗设计颇为重要。gp120作为HIV-1的包膜糖蛋白，能够诱导广泛的中和抗体反应，中和进入机体的病毒粒子，阻止病毒早期感染，所以本实验选取HIV-1 C亚型密码子优化的gp120作为免疫原进行研究。目前的疫苗研究中，腺病毒载体是较理想的病毒载体之一，具有安全性好、外源基因容纳量大、感染效率高、操作简便等优点。我们以复制缺陷型腺病毒为载体，构建了表达HIV-1 C亚型密码子优化的gp120的重组腺病毒vAd-gp120，经Western Blot方法检测到了gp120蛋白的表达。树突状细胞（DC）是已知最强的抗原呈递细胞(APC)，也是目前发现的唯一能够刺激初始型T细胞增殖的细胞。经抗原致敏的DC可通过MHC-Ⅰ、MHC-Ⅱ途径递呈抗原，并激活T细胞，从而激发体内的体液免疫和特异性细胞免疫反应。我们利用Amaxa系统将HIV-1 C亚型gp120基因转入人外周血单核细胞来源的DC，构建了以DC为载体的治疗性疫苗，并对其功能进行初步研究，发现负载gp120的DC能够显著刺激淋巴细胞的增殖、增强CD8T细胞表面活化分子CD25的表达以及促进CD8T细胞分泌++IFN-γ，为下一步DC治疗性疫苗的体内研究奠定了基础。

同源四倍体水稻雌配子体发育特性及变异胚囊研究

通过秋水仙素诱导获得同源四倍体水稻10个株系，包括6个恢复系、3个保持系和1个不育系，这些株系具有加倍的染色体组。田间观察表明10个株系具有特殊的农艺性状：茎杆变粗壮、植株颜色加深、叶片变厚、叶宽适度增加、分蘖数减少、有效分蘖的比率下降等。根尖有丝分裂鉴定表明，同源四倍体水稻10个株系具有正常的有丝分裂，观察细胞的染色体数目皆为2n=48。花粉母细胞减数分裂鉴定表明10个株系具有比较理想的减数分裂行为，后期I染色体滞后、末期I微核生成和末期II异常小孢子比率较低，能较好的完成减数分裂过程，其中后期I染色体滞后比率约为10%-20%，末期I微核生成比率约为1%-6%，末期II异常小孢子比率约为1%-8%。这提示，染色体联合和分离不规则导致三价体、单价体 和落后染色体等产生，并进一步导致在后期和末期不均横分离产生异常小孢子，这可能是同源四倍体株系结实率不高的原因之一。 同源四倍体水稻正常胚囊为蓼型，变异胚囊具有多种类型，其比率显著高于二倍体对照，变化范围为39.62%－69.85%。按变异胚囊的结构特点和形成方式，分为四种类型：退化型，结构变异型，无融合生殖型和反足细胞增殖型。退化型胚囊的平均比率为29.17%，包括小胚囊（15.04%）和完全退化胚囊（14.13%），前者仍有较小胚囊腔而后者胚囊腔缺失。结构变异胚囊包括结构缺失、结构重复和位置异常，反映了蓼型胚囊八核七细胞结构的变异，其在各株系的平均比率为18.96%。无融合生殖胚囊发生比率极低，平均比率为1.77%，类型包括反足胚和卵细胞胚。反足细胞增殖胚囊是反足细胞团频繁增殖形成，伴随上述三种变异发生使异常胚囊的多样性进一步增加，其在各株系的平均比率为10.62%。相关分析表明，同源四倍体水稻结实可能主要来自三部分：正常胚囊、正常型小胚囊和反足细胞增殖型胚囊。这三种胚囊具有相对完整的蓼型结构，可能具有较好的育性，其对结实率的贡献程度估计值分别为72.44%、15.12%、12.44%。此外，完全退化型胚囊和位置异常型胚囊对结实率分别表现出显著（-0.66）和极显著（-0.92）的负相关，这表明二者可能是结实性的抑制因素。 Ten autotetraploid strains, which include six restoring lines, three maintaining lines and a sterile line, are derived from artificial induction by colchicine treatments. Variations of agronomical traits are observed which include large organs, sturdy plants, long panicle length and deep leaf color and so on. It has been confirmed that autotetraploid strains exhibit normal chromosome behaviors in mitosis and the chromosome numbers are all 48. Moreover, abnormal chromosome behaviors are investigated in meiosis including univalent, trivalent, quatrivalent, chromosome lagging and microkernel and so on. It evaluates that the percentage of chromosome lagging in anaphase I is about 10%-20%, the percentage of microkernel in telophase I is about 1%-6% and the percentage of abnormal microspore in telophase II is about 1%-8%. In all, abnormal behaviors of chromosomes could induce univalent, trivalent and et al. and subsequently induce infertile microspore. That may be one of the causes of low seed sets in autotetraploid strains. Embryo sacs of autotetraploid strains are formed according to the Polygonum type. However, these strains exhibit variations of abnormal embryo sacs at high frequencies (39.62% - 69.85%). The variations are frequently involved in the spikelets of the main panicles and the first tillers, leading to obvious decreases of the percentages of normal embryo sacs among the strains. Four types of abnormal embryo sacs are classified basing on their different structures and origins: degenerated embryo sac (DES), structure variation (SV), apomixis (Apo) and antipodal cell proliferation (ACP). Embryo sacs of DES (29.17%) exhibit small embryo sacs (15.04%) or no embryo sac (14.13%), the former showing embryo sacs with decreased size and the latter showing no sac. Embryo sacs of AS (18.96%) include three subtypes: structure deletion, structure duplication and location variation, which suggests abnormalities of the eight nuclei, seven celled pattern of the Polygonum type. Embryo sacs of Apo (only 1.77%) include two origins of apomictic embryos: antipodal embryo and egg embryo. Embryo sacs of ACP are observed frequently (10.62%) in autotetraploid strains which subsequently increase the variations of abnormal embryo sacs. It evaluates by the Pearson’s correlation analysis that seed set is probably contributed by three origins of embryo sacs: normal embryo sacs, small embryo sacs (normal pattern) and embryo sacs of ACP. These three origins exhibit comparatively good structure of the Polygonum type and could account for seed set at a percentage of 72.44%, 15.12%, 12.44%, respectively. Moreover, the subtype of no embryo sac (NES) negatively related to seed set at the P>0.01 level (-0.92) and the subtype of location variation (LV) negatively related to seed set at the P>0.05 level (-0.66). Which suggest the two subtypes may have strong stress on seed set.

Studies on enzymological properties of antibody elicited by meso-tetra(alpha, alpha, alpha, alpha-O-phenylacetyl benzene)porphyrin

meso-Tetra (alpha, alpha, alpha, alpha-O-phenylacetyl benzene) porphyrin was used as a complete antigen to elicit monoclonal antibody 1F2 through the immunization and cell fusion techniques. McAb 1F2 obtained was demonstrated very pure by HPLC and MALDI/TOFMS. The retention time of McAb 1F2 was 2. 63 min. The subtype of McAb 1F2 was IgG2a. The relative molecular weight was 156 678. 8. When the McAb 1F2-porphyrin was formed, the maximal absorption of the porphyrin soret region had a redshift from 408 to 416 nm and hyperchromical effect, showing that the antigen-antibody combination was rigid and intense, and the abzyme constancy was high. But compared with HRP, the activity of the abzyme was only 4. 687 5 U/mg and 1. 899 % of that of HRP. Its K-m was 20. 29 mmol/L, k(cat) 396. 82 min(-1), k(cat)/K-m. 1. 955 7 X 10(4) L . mol(-1) . min(-1).

Influence of intermolecular entanglements on the glass transition and structural relaxation behaviors of macromolecules. 2. Polystyrene and phenolphthalein poly(ether sulfone)

The effect of entanglements on the glass transition and structural relaxation behaviors has been studied for polystyrene (PS) and phenolphthalein poly(ether sulfone) (PES-C) samples by fast evaporation of the solution of concentrations varying from above the overlapping concentration to far below it, and compared to the results we have studied previously in PC. It has been found that for all the polymers we have studied, in the concentrated solution region, the T-g of the samples obtained from solution are independent of the change of concentration and are very close to that of normal bulk samples, whereas in the dilute solution region the T-g of the samples decrease with the logarithm of decreasing concentration. The critical concentrations that divide the two distinct regions for the three polymers are 0.9% g/mL for PC, 0.1% g/mL for PS, and 1% g/mL for PES-C. The decrease of T-g of the samples is interpreted by the decrease of intermolecular entanglements as the isolation of polymer chains, and the entanglement of polymer chains restrained the mobility of the segments. The structural relaxation behavior of the polymers is also found to be different from that of normal bulk samples. The enthalpies of single-chain samples are lower than that of the bulk ones, which correspond to the lower glass transition temperature; the peaks are lower and broader, and the relaxed enthalpy is much lower as compared to that of bulk samples. In the three polymers we have studied, the influence of change of entanglements on both the decrease in glass transition temperature and relaxed enthalpy is the most significant for PS and the least for PES-C. It is indicated that the interactions in the flexible polymers are weak; thus, the restraint of the entanglements on the mobility of the segments plays a more important role in the flexible polymers, and the change of entanglement in the flexible polymers has a more significant influence on the physical properties.

Elliptic Object Detection Based on Shape Preserving and Active Contour

本文通过形状约束方程(组)与一般主动轮廓模型结合,将目标形状与主动轮廓模型融合到统一能量泛函模型中,提出了一种形状保持主动轮廓模型即曲线在演化过程中保持为某一类特定形状。模型通过参数化水平集函数的零水平集控制演化曲线形状,不仅达到了分割即目标的目的,而且能够给出特定目标的定量描述。根据形状保持主动轮廓模型,建立了一个用于椭圆状目标检测的统一能量泛函模型,导出了相应的Euler-Lagrange常微分方程并用水平集方法实现了椭圆状目标检测。此模型可以应用于眼底乳头分割,虹膜检测及相机标定。实验结果表明,此模型不仅能够准确的检测出给定图像中的椭圆状目标,而且有很强的抗噪、抗变形及遮挡性能。

基于形状保持主动轮廓模型长直条的检测

通过形状约束方程(组)与一般主动轮廓模型结合,将目标形状与主动轮廓模型融合到统一能量泛函模型中,提出一种形状保持主动轮廓模型。模型通过参数化水平集函数的零水平集表示某一类特定形状,不仅达到了分割即目标的目的,而且能够给出特定目标的定量描述。根据形状保持主动轮廓模型,建立一个用于长直条状目标检测的统一能量泛函模型,导出相应的Euler-Lagrange常微分方程并用水平集方法实现了长直条状区域的检测。此形状保持模型的一种特殊情况可以用于直线状地平(海天)线提取。实验结果表明,该模型不仅能够准确地检测出给定图像中的长直条状区域而且有很强的抗噪、抗变形及遮挡性能。

柔索驱动三自由度球面并联机构运动学与静力学研究

柔索驱动并联机器人采用柔索代替连杆作为机器人的驱动元件 ,它结合了并联结构和柔索驱动的优点 .文章提出了一种新型带有约束机构的并联柔索驱动机器人 ,采用四根柔索驱动 .由于约束机构的引入 ,机器人可实现在空间的三维转动 .介绍柔索驱动并联机器人的机构构型 ,给出了位姿逆解 ,建立了静力平衡方程和运动学方程 ,讨论了柔索拉力的确定方法 .研究结果证明在加入了约束机构后 ,柔索机器人可以实现更多的运动形式 ,这就为更广泛的应用柔索驱动成为可能

深海机器人推进电机系统的混沌控制

深海机器人推进电机系统中出现的混沌现象,直接影响深海机器人稳定性、可靠性和安全性.采用自适应控制技术对其混沌行为加以控制,对该方法的可行性和有效性进行了证明.设计和构造了易于工程实施的混沌控制器,用于深海机器人推进电机系统混沌控制.仿真实验表明,推进电机系统在自适应控制器的作用下可迅速脱离混沌状态,并进入持续稳定状态,控制效果明显.可以为深海机器人推进电机系统中可能出现的混沌运行行为提供控制策略和抑制预案,有利于混沌控制嵌入软件的开发,确保深海机器人稳定、可靠和安全地运行,具有一定的实用价值.

成批生产计划调度的集成建模与优化

针对多品种批量生产类型,建立了调度约束的生产计划与调度集成优化模型。模型的目标函数是使总调整费用、库存费用及生产费用之和最小,约束函数包括库存平衡约束和生产能力约束,同时考虑了调度约束,即工序顺序约束和工件在单机上的加工能力约束,保证了计划可行性。该模型为两层混合整数规划模型,对其求解综合运用了遗传算法和启发式规则,提出了混合启发式求解算法。最后,针对某机床厂多品种批量生产类型车间进行了实例应用,对车间零件月份作业计划进行分解,得到各工段单元零件周作业计划,确定了零件各周生产批量与投产顺序。

黄骅坳陷港西开发区河流相储层构型与剩余油分布模式研究

In the production tail of oilfield, water-cut is very high in thick channel sand oil reservoir, but recovery efficiency is relative low, and recoverable remaining oil reserves is more abundant, so these reserves is potential target of additional development. The remaining oil generally distributed with accumulation in certain areas, controlled by the reservoir architecture that mainly is the lateral accretion shale beddings in the point bar, so the study of reservoir architecture and the remaining oil distribution patterns controlled by architecture are very significant. In this paper, taking the Minghuazhen formation of Gangxi oilfield as a case, using the method of hierarchy analysis, pattern fitting and multidimensional interaction, the architecture of the meandering river reservoir is precisely anatomized, and the remaining oil distribution patterns controlled by the different hierarchy architecture are summarized, which will help to guide the additional development of oil fields. Not only is the study significant to the remaining oil forecasting, but also it is important for the theory development of reservoir geology. With the knowledge of sequence correlation and fluvial correlation model, taking many factors into account, such as combination of well and seismic data, hierarchical controlling, sedimentary facies restraint, performance verification and 3-D closure, an accurate sequence frame of the study area was established. On the basis of high-resolution stratigraphic correlation, single layer and oil sand body are correlated within this frame, and four architecture hierarchies, composite channel, single channels, point bars and lateral accretion sandbody are identified, The result indicates that Minghuazhen Formation of Gangxi oilfield are dominated by meandering river deposition, including two types of channel sandbodies, narrow band and wide band channel sandbody, and each of them has different characteristics of facies variation laterally. Based on the identification of composite channel, according to the spatial combination patterns and identified signs of single channel, combined with channel sandbody distribution and tracer material data, single channel sandbodies are identified. According to empirical formula, point-bar scales of the study area are predicted, and three identification signs are summarized, that is, positive rhythm in depositional sequence, the maximum thick sand and near close to the abandoned channel, and point bars are identified. On the basis of point bar recognition, quantitative architecture models inner point bar are ascertained, taking the lateral accretion sand body and lateral accretion shale beddings in single well as foundation, and quantitative architecture models inner point bar as guidance, and result of tracer material data as controlling, the the lateral accretion sand body and lateral accretion shale beddings are forecasted interwell, so inner architecture of point bar is anatomied. 3-D structural model, 3-D facies model and 3-D petrophysical properties models are set up, spatial distribution characteristics of sedimentary facies and petrophysical properties is reappeared. On the basis of reservoir architecture analysis and performance production data, remaining oil distribution patterns controlled by different hierarchy architecture units, stacked channel, single channel and inner architecture of point bar, are summarized, which will help to guide the additional development of oil fields.

抑郁行为及不同抑郁亚型的多巴胺机制研究

The conceptualization of “depression” as a heterogenous disease has been widely accepted by most researchers. However, controlled experiments are rather sparse. To date, most studies demonstrated that animals with helplessness, a widely recognized behavioral index of “depression” also show varied comobidity expressions of other emotional behaviors, such as hightened or lightened anxiety level compared with controls. This means that distinct subtypes of “depression” may exist, in which different neural mechanisms may play roles. The present study aims to explore the possibility of behaviorally categorizing two depressive subtypes, referred as anxious helplessness and non-anxious helplessness, respectively. Then, by using RT-PCR, the dopamine D1, D2, D3 receptors mRNA expressions in medial prefrontal cortex (mPFC) and nucleus accubems (NAc) were quantified. The main findings are described as belows: 1. Uncontrollable shock could readily induce helpless behavior in shocked animals as a whole but with salient individual differences. Prior inescaoable shock induces subsuquent helplessness in approximately 40% shocked animals, while the other animals showed no sign of helpless expression, and were classified as non-helplessness. 2. Among helpless animals, the “subtype” of anxious helpless and non-anxious helpless could be identified according to the anxiety level evaluated by elevated plus maze. 3. D3 receptors mRNA expressions in the mPFC and NAc were increased in stressed animals after uncontrollable shock treatment. At the meanwhile, significant lower expressions of D2 receptors in the mPFC and NAc, and much lower expressions of D1 receptors in the mPFC were found in rats that did not become helpless after stress. In contrast, no significant difference between helpless and control animals was found in D1/D2 receptors mRNA expressions. 4. Based on above mentioned results, the up-regulation of D3 receptors in the mPFC and NAc may reflect a generalized effect of exposure to uncontrollalbe shock. While the down-regulation of D1\D2 receptors in the mPFC and decreased expression of D2 receptors in the NAc may be associated with adaptive or protective mechnisms which protecting animals from helplessness after uncontrollable shock treatment. 5. Futhermore, a significant negative relationship was found between anxiety level and D1 receptors expressions in the mPFC in helpless animals. Compared to the non-anxious helpless and control rats, the D1 receptors mRNA of anxious helpless rats were down-regulation in the mPFC. The present study indicated that the D1 dopamine receptor gene is associated with co-morbid depression and anxiety.