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.     

A numerical study of dynamic behaviors of a unitized regenerative fuel cell during gas purging

The gas purging states affect electricity output and energy storage capacity of unitized regenerative fuel cells. In this study, a model of unitized regenerative fuel cell is established. Cell voltages and operating temperatures influences on the dynamic distribution of thermal fluid during purging process and the discharge of residual liquid water in electrolytic cell mode are investigated. The motivation of the present study is better understanding the gas purging characteristics and its effect on reaction behaviors of unitized regenerative fuel cells. Simulation results reveal a significant influence of purging gas temperature on the water flooding and a great effect of operating voltage on the water diffusion. The operating temperature of electrolytic cell model almost has little effect on purging results at different cell temperature and the same purging gas temperature. When the purging gas temperature is changed, higher temperatures of cell and purging gas facilitate liquid water discharging out from the cell regions. In cell water flooding situation, when having large liquid content, the purging gas has little effects on the water expelling process.     

A charge sharing–based switching scheme for SAR ADCs

This paper presents an energy-efficient switching scheme for successive approximation register (SAR) analogue-to-digital converter (ADC). The proposed scheme employs charge recycling method to keep the capacitor arrays free of transitional energy between bit generations except reset phase. In comparison with the conventional switching scheme, the proposed one achieves 100% transitional energy saving without considering reset phase. In addition, configuration of a 10-bit SAR ADC shows that the proposed switching scheme reduces the capacitor area by 25% compared with the conventional switching scheme.     

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。     

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。     

Effects of welding parameters and welding sequence on residual stress and distortion in Al6061-T6 aluminum alloy for T-shaped welded joint

The distribution of temperature and then the distribution of residual stress and distortion in the stiffened aluminum alloy Al6061-T6 plates under the metal inert gas (MIG) welding process were investigated by three dimensional thermo-mechanical coupled finite element model using Ansys software. The properties of materials were considered temperature-dependent and the filler metal was added to the workpiece by the element birth and death technique. In three modes of current, two different speeds and two various sequences, the distribution of residual stress and distortion were calculated and analyzed. The results showed that increase in welding speed decreased the vertical deflection in the plate, transverse shrinkage and angular distortion of plate and the lateral deflection of stiffener, but increased the maximum longitudinal tensile stress in the plate and stiffener. Furthermore, increase in current increased the residual stress and deformation in the plate and stiffener, and the change in the welding sequence changed the distribution of the distortion in the plate and the stiffener without significant change in the distribution of the longitudinal residual stress.