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.     

HPLC法测定伐地昔布的有关物质

建立高效液相色谱法检测伐地昔布的有关物质.采用α-酸性糖蛋白键合硅胶的手性色谱柱,以乙腈-醋酸铵缓冲液(0.01 mol/L,pH4.0)作为流动相进行梯度洗脱,检测波长215 nm.伐地昔布与5个已知杂质均可达到基线分离,在各自浓度范围内线性关系良好,平均回收率95.13％～99.77％,RSD1.10％～1.77％,检测限0.017 9～0.060 1 μg/mL,定量限0.059 7～0.200 0 μg/mL.方法专属性强,准确度好,灵敏度高,可为伐地昔布的质量控制及后续分离提纯和帕瑞昔布的质量控制提供指导.     

S120在薄带铸轧主轧机控制系统上的应用

薄带铸轧生产线设备紧凑,生产工艺更加低耗高效,是钢铁工业流行发展的趋势.SINAMICS S120控制系统应用在薄带铸轧主轧机传动系统上,可以根据需求灵活增添功能配置.通过介绍该生产线工艺流程,突出S120传动控制系统的优势.此外,针对S120控制系统设计和应用,尤其是负荷平衡控制、SINAMICS Link通讯、安全转矩关断等特殊应用的实现进行了详细的阐述.     

空间曲线的特征识别与高质量非均匀采样

为避免传统均匀采样方法因忽视曲线重要特征而生成不理想的采样结果,获得给定数量且由特征点和辅助点组成的采样点序列,提出基于特征识别的高质量空间曲线非均匀采样方法.首先使用抛物线插值法得到曲线上所有曲率极大值点和挠率极大值点的近似位置,经筛选后产生特征点,以更好地抓住空间曲线的轮廓特征.然后定义基于弧长、曲率和挠率加权组合的特征函数,并以此自适应地选取曲线上的辅助点.与3种主流采样方法比较的实验结果表明,该方法能够获得更高质量的采样结果且具有更好的实用性,从而进一步改善空间曲线的B样条拟合效果.     

Effects of multiple freeze–thaw cycles on meat quality,nutrients, water distribution and microstructure in bovine rumen smooth muscle

This study investigated the effect of 5 freeze–thaw cycles (freezing at −18°C for 12 h and then thawing at 4°C for approximately 12 h) on the meat quality, proximate composition, water distribution and microstructure of bovine rumen smooth muscle (BSM). As the number of freeze–thaw cycles increased, BSM pH, shear force, water content and protein content decreased by 3.06%, 35.50%, 14.49% and 21.11%, respectively, whereas BSM thawing loss, cooking loss, pressing loss, total aerobic count (TAC), ash content and fat content increased by 108.12%, 47.75%, 78.33%, 90.99%, 105% and 35.20%, respectively. The freeze–thaw cycles resulted in greater protein and lipid oxidation, as evidenced by a 36.46% reduction in the sulfhydryl content and a 209.06% and 338.46% increase in the carbonyl and malondialdehyde contents, respectively. Ice crystal formation disrupted the structural integrity of the muscle tissue. Low-field nuclear magnetic resonance results showed that the freeze–thaw cycles prolonged the relaxation times (T_2b, T₂₁ and T₂₂), indicating that immobile water shifted to free water, and consequently, free water mobility increased. After 3 freeze–thaw cycles, the decline in shear force slowed, the increase in thawing loss became accelerated, and the TAC approached the domain value (6 log colony-forming units/g). Therefore, the number of freeze–thaw cycles of smooth muscle during transport, storage and distribution should be controlled to 3 or fewer. The current results provide a theoretical basis and data support for the further utilisation and culinary processing of smooth muscle.     

CDBN‑IKELM的轴承变工况故障诊断方法

针对现有方法在轴承变工况方面存在的诊断精度低、人工提取特征不充分等问题，提出了基于卷积深度置信网络（convolutional deep belief network，简称CDBN）与改进核极限学习机 （improved Kernel?based extreme learning machine，简称IKELM）的滚动轴承故障智能识别方法。首先，由卷积深度置信网络对原始信号内的故障特征进行深层自适应提取；其次，利用等距特征映射对提取的多维特征进行降维，去除冗余特征信息；然后，采用改进的核极限学习机对特征进行分类，使用粒子群（particle swarm optimization， 简称PSO）对模型重要参数进行优化，实现滚动轴承变工况下的故障识别；最后，将所提方法应用于不同工况下多种轴承故障的诊断。实验结果表明，该方法能够智能有效地识别变工况的轴承故障，诊断结果优于已有的智能故障诊断方法。     

Changes in polyphenol fractions and bacterial composition after in vitrofermentation of apple peel polyphenol by gut microbiota

The changes in polyphenol fractions after in vitro fermentation of apple peel polyphenol (APP) by gut microbiota as well as the effects of APP on the growth, pH value, short-chain fatty acids (SCFAs) production and intestinal flora composition of gut microbiota fermentation were firstly explored in this study. The relative abundance of Lactobacillus in the APP group was 49.55% with an increment of 49.40% compared with the blank group, while that of Bifidobacterium was 13.32%. Moreover, the flora produced 37.093 ± 0.478 mM of SCFAs including acetic acid, propionic acid, butyric acid and valeric acid during the process of fermentation, thus reduced the environmental pH value. In addition, polyphenol fractions in APP were altered by gut microbiota fermentation to some extent, for example, glycosides were hydrolysed to aglycones. These findings suggested APP as a potential prebiotic agent to alleviate the disorder of intestinal flora.