Targeted editing of intronic-splicing silencer enhancement of SMN2 Exon 7 inclusion by CRISPR/Case 9

Spinal muscular atrophy (SMA) is an autosomal recessive hereditary neuromuscular disease. Exon 7 and 8 of survival of motor neuron 1 (SMN1) gene or only exon 7 homology deletion leads to the failure to produce a full-length SMN gene. The copy number of SMN2 gene with high homology of SMN1 affects the degree of disease and was the target gene for targeting therapy, in which splicing silencer in intron 7 was the key to suppress the inclusion of exon 7. In this study, we projected to use CRISPR/Case 9 for the targeted editing of intronic-splicing silencer (ISS) sequence to promote the inclusion of SMN2 exon 7 and increase the production of SMN2 full-length (FL) gene expression. It happens that there was a protospacer adjacent motif (PAM) at one end of the ISS sequence according to the design of sgRNA. The recombinant vector of sgRNA HSMN2 CRISPR/Case 9 was constructed and transfected into HEK293 cells. Sequencing results showed that the ISS sequence could be edited accurately and targeting in the predicted direction, in which deleting small fragments, inserting small amounts and mutation. Quantitative analysis of RT-PCR products by restriction enzyme of DdeI digestion showed that the FL of SMN2 increased by 8% (P < 0.05). In the primary cultured chondrocytes of SMA mice, in which sgRNA HSMN2 CRISPR/Case9 recombinant vector transfection could increase the SMN2 FL gene by 23% (P < 0.05) and significantly improve SMN protein levels (P < 0.05). CRISPR/Case 9 is an effective tool for gene editing and therapy of hereditary diseases, but it is rarely reported in the treatment of SMA diseases. This study shows that CRISPR/Case 9 was first used for the precision target of ISS sequence editing, which can effectively promote the production of SMN2 FL gene expressions, in which there was an important clinical reference value.     

Exploration of GH94 Sequence Space for Enzyme Discovery Reveals a Novel Glucosylgalactose Phosphorylase Specificity

The substantial increase in DNA sequencing efforts has led to a rapid expansion of available sequences in glycoside hydrolase families. The ever-increasing sequence space presents considerable opportunities for the search for enzymes with novel functionalities. In this work, the sequence-function space of glycoside hydrolase family 94 (GH94) was explored in detail, using a combined approach of phylogenetic analysis and sequence similarity networks. The identification and experimental screening of unknown clusters led to the discovery of an enzyme from the soil bacterium Paenibacillus polymyxa that acts as a 4-O-β-d -glucosyl-d -galactose phosphorylase (GGalP), a specificity that has not been reported to date. Detailed characterization of GGalP revealed that its kinetic parameters were consistent with those of other known phosphorylases. Furthermore, the enzyme could be used for production of the rare disaccharides 4-O-β-d -glucosyl-d -galactose and 4-O-β-d -glucosyl-l -arabinose. Our current work highlights the power of rational sequence space exploration in the search for novel enzyme specificities, as well as the potential of phosphorylases for rare disaccharide synthesis.     

Image sequence-based risk behavior detection of power operation inspection personnel

A novel image sequence-based risk behavior detection method to achieve high-precision risk behavior detection for power maintenance personnel is proposed in this paper. In this method, the original image sequence data is first separated from the foreground and background. Then, the free anchor frame detection method is used in the foreground image to detect the personnel and correct their direction. Finally, human posture nodes are extracted from each frame of the image sequence, which are then used to identify the abnormal behavior of the human. Simulation experiment results demonstrate that the proposed algorithm has significant advantages in terms of the accuracy of human posture node detection and risk behavior identification.     

Facial expression recognition of intercepted video sequences based on feature point movement trend and feature block texture variation

Facial Expression Recognition (FER) is an important subject of human–computer interaction and has long been a research area of great interest. Accurate Facial Expression Sequence Interception (FESI) and discriminative expression feature extraction are two enormous challenges for the video-based FER. This paper proposes a framework of FER for the intercepted video sequences by using feature point movement trend and feature block texture variation. Firstly, the feature points are marked by Active Appearance Model (AAM) and the most representative 24 of them are selected. Secondly, facial expression sequence is intercepted from the face video by determining two key frames whose emotional intensities are minimum and maximum, respectively. Thirdly, the trend curve which represents the Euclidean distance variations between any two selected feature points is fitted, and the slopes of specific points on the trend curve are calculated. Finally, combining Slope Set which is composed by the calculated slopes with the proposed Feature Block Texture Difference (FBTD) which refers to the texture variation of facial patch, the final expressional feature are formed and inputted to One-dimensional Convolution Neural Network (1DCNN) for FER. Five experiments are conducted in this research, and three average FER rates 95.2%, 96.5%, and 97% for Beihang University (BHU) facial expression database, MMI facial expression database, and the combination of two databases, respectively, have shown the significant advantages of the proposed method over the existing ones.     

吐哈盆地葡北地区“两宽一高”地震勘探技术及应用效果

吐哈盆地台北凹陷圈闭落实的准确度和储层分布预测的精度一直是制约该区油气发现的关键因素。通过可控震源“两宽一高”地震采集、OVT域时间偏移、地质导向深度偏移初始模型建立及各向异性深度偏移迭代处理，在地震资料的信噪比、频带宽度和分辨率等方面明显提高的基础上，以高精度层序地层学为指导，基于高分辨率地震资料与钻井、测井资料的综合分析，识别出侏罗系6个三级层序，在三级层序格架内将研究区七克台组底部和三间房组顶部砂体识别为两个连续上升半旋回四级层序；通过构造几何属性分析及精细构造分析，揭示了本区“四带、两凹”的构造格局及形成机制；以宽频带多种反演技术为手段，对含油气层段的七克台组和三间房组砂体分布进行了预测；在综合构造分析、储层分析和油藏成藏规律分析的基础上，提出本区油气沿构造带脊线聚集，断裂系统是油气运移的优势路径，七克台组、三间房组储层在葡北构造带发育，向玉果构造带和胜北次凹减薄尖灭，葡北构造带与玉果构造带结合部位的，具有鼻状构造背景的岩性圈闭是获得油气突破的有利地区。     

华莱士瓜采后丝状真菌的分离及rDNA ITS区序列分析

从采后贮藏期间自然发病的华莱士瓜上分离到12株丝状真菌,将分离菌经纯化后回接到健康无损的华莱士瓜上,发病特征与分离发病果实一致,确定其为华莱士瓜的病原菌。结合形态学观察和rDNA内转录间隔区(internal transcribed spacer,ITS)序列分析,结果表明:分离到1株黑附球菌(Epicoccum nigrum)、1株草酸青霉菌(Penicillium oxalicum)、2株链格孢属(Alternaria Nees ex Wallr.)真菌、3株曲霉属(Aspergillus Micheli ex Fr.)真菌和5株镰刀属(Fusarium LK. ex Fr.)真菌。其中,黑附球菌(E. nigrum)为甜瓜中首次发现的病原菌,而草酸青霉菌(P. oxalicum)、细极链格孢菌(Alternaria tenuissima)、泡盛曲霉菌(Aspergillus awamori)、木贼镰刀菌(Fusarium equiseti)、轮状镰刀菌(F. verticillioides)和燕麦镰刀菌(F. avenaceum)分别为甜瓜上分离到的青霉属、链格孢属、曲霉属和镰刀属的新菌株。     

浅埋煤层群采场周期来压顶板结构及支架载荷

现场勘查发现,神府矿区浅埋煤层群间隔岩层厚度大多在15～45 m时,间隔岩层易存在单一关键层结构。文章通过现场实测得出了浅埋煤层群开采周期来压的基本特征:来压步距减小、强度增大、煤壁片帮严重,动载现象、台阶下沉现象和大、小周期来压现象明显。物理相似模拟实验揭示了间隔岩层关键层与上煤层已扰动关键层同步与非同步破断的大、小周期来压机理,发现了间隔岩层关键层与已扰动关键层共同形成的"梁-拱-壳"结构形态,给出了上煤层已扰动关键层结构形态的判别方法,得出了浅埋煤层群开采顶板周期破断可形成"台阶-台阶"、"砌体-台阶"、"砌体-砌体"的基本结构。综合建立了浅埋煤层群开采间隔岩层周期破断结构与力学模型,给出了支架受动静载荷作用的工作阻力计算方法,并得到了现场验证。     

基于修正等速运动规律的自由曲面多轴加工刀轴序列优化方法

针对自由曲面多轴加工中起始位置处刀具相对工件速度、加速度不为零导致的一定程度刚性冲击现象,提出了一种基于修正等速运动规律的刀轴序列优化方法,通过控制工件坐标系中决定刀具姿态的两个约束角,使之变化遵循修正等速运动规律,获得了切削行上平稳变化的刀轴序列,避免了由于起始位置处刀具相对工件速度、加速度不为零导致刀具不能精准到达后续加工位置的现象。算例分析表明,该方法能产生平稳且光滑的刀轴序列,可一定程度改善机床运动性能,提高加工表面质量。     

银额盆地拐子湖凹陷北部烃源岩特征及控制因素

针对拐子湖凹陷勘探程度较低、有效烃源岩分布及控制因素不明确的问题,运用有机地球化学、沉积层序及构造演化分析方法,对温1井烃源岩特征及主控因素展开研究。研究结果表明:研究区烃源岩有机质丰度整体较低,受沉积层序、构造综合影响,高位体系域有利于有机质富集;有机质类型为腐泥腐殖型和腐殖型;有机质热演化程度差异较大,富泥质岩类的发育可促进早期的热演化进程,构造的后期抬升使局部生烃时间大幅缩短,故有机质成熟度从未成熟至高成熟均有分布;烃源岩展布受断裂及沉积演化控制明显。该研究可为拐子湖凹陷北部下一步勘探提供参考。