Complex reorganization and predominant non-homologous repair following chromosomal breakage in karyotypically balanced germline rearrangements and transgenic integration

We defined the genetic landscape of balanced chromosomal rearrangements at nucleotide resolution by sequencing 141 breakpoints from cytogenetically interpreted translocations and inversions. We confirm that the recently described phenomenon of 'chromothripsis' (massive chromosomal shattering and reorganization) is not unique to cancer cells but also occurs in the germline, where it can resolve to a relatively balanced state with frequent inversions. We detected a high incidence of complex rearrangements (19.2%) and substantially less reliance on microhomology (31%) than previously observed in benign copy-number variants (CNVs). We compared these results to experimentally generated DNA breakage-repair by sequencing seven transgenic animals, revealing extensive rearrangement of the transgene and host genome with similar complexity to human germline alterations. Inversion was the most common rearrangement, suggesting that a combined mechanism involving template switching and non-homologous repair mediates the formation of balanced complex rearrangements that are viable, stably replicated and transmitted unaltered to subsequent generations.     

Heritable and inducible genetic interference by double-stranded RNA encoded by transgenes

Double-stranded RNA interference (RNAi) is an effective method for disrupting expression of specific genes in Caenorhabditis elegans and other organisms. Applications of this reverse-genetics tool, however, are somewhat restricted in nematodes because introduced dsRNA is not stably inherited. Another difficulty is that RNAi disruption of late-acting genes has been generally less consistent than that of embryonically expressed genes, perhaps because the concentration of dsRNA becomes lower as cellular division proceeds or as developmental time advances. In particular, some neuronally expressed genes appear refractory to dsRNA-mediated interference. We sought to extend the applicability of RNAi by in vivo expression of heritable inverted-repeat (IR) genes. We assayed the efficacy of in vivo-driven RNAi in three situations for which heritable, inducible RNAi would be advantageous: (i) production of large numbers of animals deficient for gene activities required for viability or reproduction; (ii) generation of large populations of phenocopy mutants for biochemical analysis; and (iii) effective gene inactivation in the nervous system. We report that heritable IR genes confer potent and specific gene inactivation for each of these applications. We suggest that a similar strategy might be used to test for dsRNA interference effects in higher organisms in which it is feasible to construct transgenic animals, but impossible to directly or transiently introduce high concentrations of dsRNA.     

Purification and cloning of amyloid precursor protein beta-secretase from human brain

Proteolytic processing of the amyloid precursor protein (APP) generates amyloid beta (Abeta) peptide, which is thought to be causal for the pathology and subsequent cognitive decline in Alzheimer's disease. Cleavage by beta-secretase at the amino terminus of the Abeta peptide sequence, between residues 671 and 672 of APP, leads to the generation and extracellular release of beta-cleaved soluble APP, and a corresponding cell-associated carboxy-terminal fragment. Cleavage of the C-terminal fragment by gamma-secretase(s) leads to the formation of Abeta. The pathogenic mutation K670M671-->N670L671 at the beta-secretase cleavage site in APP, which was discovered in a Swedish family with familial Alzheimer's disease, leads to increased beta-secretase cleavage of the mutant substrate. Here we describe a membrane-bound enzyme activity that cleaves full-length APP at the beta-secretase cleavage site, and find it to be the predominant beta-cleavage activity in human brain. We have purified this enzyme activity to homogeneity from human brain using a new substrate analogue inhibitor of the enzyme activity, and show that the purified enzyme has all the properties predicted for beta-secretase. Cloning and expression of the enzyme reveals that human brain beta-secretase is a new membrane-bound aspartic proteinase.     

水合锰(Ⅱ)结构的量子化学和ABEEM/MM研究

应用新一代可极化分子力场——原子-键电负性均衡浮动电荷分子力场ABEEM/MM,结合精密量子化学方法, 构建了精确的Mn²⁺-H₂O 相互作用的势能函数, 确定了相关参数.将该势能函数用于计算[Mn(H₂O)_n]²⁺(n=1~12)的结构和结合能, 得到了与量子化学一致的结果. 进一步对Mn²⁺水溶液进行ABEEM/MM 动力学模拟, 得到的Mn²⁺–O 径向分布函数的第一和第二最高峰分别处于0.218 和0.435 nm 处, 积分得到第一和第二水合层的配位水分子数分别为7.03 和17.74; 对于O–Mn²⁺–O 角度分布函数, 其第一和第二最高峰分别位于80°和140°附近, 这些结果与实验和其他理论方法的结果有很好的一致性. Mn²⁺的极化作用使得第一水合层中水分子的键长明显增长, 键角明显减小; 而Mn²⁺对第二水合层及外层水分子的结构影响较小. 分析体系的电荷分布表明, 与ABEEM-7P 纯水相比, Mn²⁺水溶液中参与形成氢键的氢原子和孤对电子的电荷变化较大, 且Mn²⁺和其邻近的水分子间存在明显的电荷转移.     

吸收对对称性一维三元光子晶体能带的影响

利用复折射率及传输矩阵理论,研究了光子晶体的吸收对对称性一维三元光子晶体能带及透射峰的影响,研究表明:在反射波中,禁带的反射率随消光系数的增加而迅速降低,当k增加到0.005时,禁带边缘模糊,不存在明显的禁带;在透射波中,随着消光系数的增大,禁带边缘逐渐模糊,当k增加为0.003时,透射率降为0.35;光子晶体的消光系数对禁带内透射峰的透射率有着明显的影响,当k为0.001时,透射率下降到0.15,随着消光系数的增加,透射峰的半峰全宽随之增加,但对透射峰的中心波长没有影响.     

基于抗差估计的变形监测基准网坐标系统转换

为监测地下煤炭开采对某矿工业广场内主、副井筒的影响,布设了由10个点构成的GPS基准网,从而为由54个监测点构成的井筒监测网提供在矿区坐标系统下的变形分析基准;在求解坐标转换参数时,为克服转换基准点中存在的位移对求参的影响,提出了采用抗差估计理论求参的模型及数据处理流程,并建立转换后GPS网的质量评价模型。根据对GPS基准网的处理结果,采用抗差估计求参,有利于保留GPS技术高精度的特点。     

轧制计划的优化模型及其应用

对于钢铁企业热轧生产控制计算机系统中的实际轧制计划初选问题,即板坯库倒垛问题,提出了一个新的非线性０－１整数规划数学模型,通过实际应用表明,该数学模型与原有的启发式经验模型相比,效果显著。     

甘氨酸在电沉积铁铬合金中的作用

研究不同浓度甘氨酸对电沉积铁铬合金过程的影响,分析了甘氨酸在电沉积过程中的作用.加入甘氨酸后,可得到优良的镀层,而不含甘氨酸时,镀层中夹杂铬的化合物,造成镀层质量低劣.甘氨酸浓度为0.4～0.8 mol·L-1时,镀层中铬含量较稳定.甘氨酸的加入使得阴极极化增加,电位负移.在此镀液中,甘氨酸与Fe(Ⅱ)络合,使其析出电位负移,达到铁铬共沉积的目的.     

亚伯拉罕·棣莫弗的概率思想与正态概率曲线

目的系统地探讨和分析正态概率曲线的发现过程及棣莫弗(Abraham De Moivre,1667- 1754)概率思想的创新点。方法历史分析和文献考证。结果棣莫弗在前人的基础上,经过10 余年的努力最终导出了正态概率曲线的数学表达式。结论正态分布是概率论中最重要的分布, 其发现揭示了棣莫弗概率思想的发展过程及其对概率论发展所做的奠基性工作。     

一种碳酸盐岩流体替换新方法

针对碳酸盐岩的基质矿物类型非单一且孔隙结构复杂多变的特点,提出一种适用于碳酸盐岩的流体替换方法。以常规Gassmann方程为基础,采用流体因子分析的基质矿物体积模量反演方法和碳酸盐岩岩石物理模型的孔隙结构反演方法,得到岩石基质矿物的体积模量,并考察复杂孔隙结构的影响,实现碳酸盐岩的流体替换。岩石物理测试分析数据和实际测井资料的计算结果均证明了新方法的正确性和有效性。