不同批次的重组诱饵型受体创新药物RC28-E1与RC28-E2对视网膜新生血管的药效学比较及机制

目的探讨不同批次的重组诱饵型受体创新药物RC28-E1与RC28-E2对视网膜新生血管的药效学比较及作用机制。方法选取健康清洁级4日龄C57BL/6J幼鼠60只，应用随机数字表法随机分为正常对照组、血管内皮生长因子（VEGF）＋成纤维细胞生长因子2（FGF2）组、VEGF＋FGF2＋RC28-E1组、VEGF＋FGF2＋RC28-E2组、VEGF＋FGF2＋conbercept组、VEGF＋FGF2＋FGF trap组，每组10只。取各组视网膜组织块构建培养体系，加入用饥饿培养基配置的相应因子和药物刺激，正常对照组加饥饿培养基。将各组视网膜组织块进行植物凝集素（Isolectin B4）染色并拍照，计算各组单位血管长度下视网膜血管顶细胞分叉的数量。另选96只健康清洁级7日龄C57BL/6J幼鼠，应用随机数字表法随机分为正常对照组、氧诱导视网膜病变（OIR）模型对照组、OIR＋RC28-E1组、OIR＋RC28-E2组、OIR＋conbercept组和OIR＋FGF trap组，每组16只。正常对照组在常氧下饲养10 d，其余各组在高氧环境中饲养5 d后在常氧状态下再饲养5 d。各组取17日龄鼠视网膜进行Isolectin B4染色并拍照，用计算机软件分析视网膜相对无灌注区面积和新生血管像素。Western blot法检测OIR实验各组视网膜中VEGF和FGF2的蛋白水平。最后，在VEGF和FGF2刺激下的视网膜血管内皮细胞（RF/6A）中，检测给予RC28-E1、conbercept以及FGF trap处理后，细胞的MEK-细胞外调节蛋白激酶（Erk）、蛋白激酶C（PKC）以及蛋白激酶B（Akt）信号传导通路的活性变化。结果视网膜组织块研究结果显示，VEGF＋FGF2＋RC28-E1组、VEGF＋FGF2＋RC28-E2组、VEGF＋FGF2＋conbercept组和VEGF＋FGF2＋FGF trap组中单位血管长度的顶细胞分叉数目均明显少于VEGF＋FGF2组，差异均有统计学意义（均P〈0.001），其中RC28-E1组与RC28-E2组的血管顶细胞分叉数量相似，差异无统计学意义（P＝0.15），但均明显少于VEGF＋FGF2＋conbercept组和VEGF＋FGF2＋FGF trap组，差异均有统计学意义（均P〈0.001）。OIR模型对照组无灌注区相对面积明显大于各药物干预组，差异均有统计学意义（均P〈0.05），OIR＋RC28-E1组与OIR＋RC28-E2组视网膜无灌注区相对面积的比较，差异无统计学意义（P＝0.17），但二者明显小于OIR＋conbercept组和OIR＋FGF trap组，差异均有统计学意义（均P〈0.05）。各干预组新生血管的相对像素值明显低于OIR模型对照组，差异均有统计学意义（均P〈0.001），OIR＋RC28-E1组视网膜新生血管的相对像素值明显低于VEGF＋FGF2＋conbercept组和VEGF＋FGF2＋FGF trap组，差异均有统计学意义（均P〈0.05），但与OIR＋RC28-E2组相比，差异无统计学意义（P＝0.39）。Western blot结果显示，OIR小鼠视网膜中VEGF和FGF2的蛋白表达均较正常对照组显著上调，差异均有统计学意义（均P〈0.001），而RC28-E1和RC28-E2可使其降至正常水平。VEGF和FGF2可诱导RF/6A细胞中的MEK-Erk通路活性增强，而RC28-E1可显著抑制这一通路的过度激活。结论RC28-E1和RC28-E2可在初生小鼠视网膜组织块中抑制血管生成，并减少OIR小鼠模型中视网膜的无灌注区，减轻新生血管生成。药理批次和中试批次的RC28-E在体内外模型中的效果相当，稳定性可靠，且优于临床同类药物conbercept和FGF trap。RC28-E1可能是通过抑制视网膜血管内皮细胞中MEK-Erk通路的过度激活而达到抑制病理性新生血管生成的效果的。

Construction and analysis of a subtracted cDNA library of Betula platyphylla female inflorescence

Female inflorescence of Betula platyphylla was sampled at an interval of eachtwo days to analyze the background of gene expression in floral phase. On the basis of SMARTstrategy, the driver cDNA was obtained from total RNA of the last sample and the tester cDNA wasfrom that of the others by RT-PCR which were subsequently used to construct a subtracted cDNAlibrary. The result of the ESTs (expression sequence tags) blastX showed that the genes in thesubtracted cDNA library could be mainly clustered into 5 groups related to metabolism,transportation and signal transduction, cell cycle, stress response, and regulation. Therelationship between gene expression and development was also discussed.

Automatic Identification of Axis Orbit Based on Both Wavelet Moment Invariants and Neural Network

Axis orbit is an important characteristic to be used in the condition monitoring and diagnosis system of rotating machine. The wavelet moment has the invariant to the translation, scaling and rotation. A method, which uses a neural network based on Radial Basis Function (RBF) and wavelet moment invariants to identify the orbit of shaft centerline of rotating machine is discussed in this paper. The principle and its application procedure of the method are introduced in detail. It gives simulation results of automatic identification for three typical axis orbits. It is proved that the method is effective and practicable.