利用CRISPR/Ca9技术靶向编辑芥蓝BoaZDS

以芥蓝(Brassicaoleraceavar.alboglabra)为材料,以ζ–胡萝卜素脱氢酶(ζ-Carotene desaturase,ZDS)基因为目标基因,建立其CRISPR/Cas9基因组编辑体系。在BoaZDS的编码区近5′端选择靶位点,构建了CRISPR/Cas9表达载体,通过农杆菌介导的遗传转化方法获得了19个芥蓝转基因阳性植株,Sanger测序分析发现其中13株成功突变,CRISPR/Cas9载体在芥蓝上的突变效率为68.42%,且所有突变植株均表现出明显的白化表型。     

Effect of Kechuanning on airway remodeling and protein level of p-ERK1/2 in lung tissues of asthmatic rats induced by virus

AIM:To investigate the effect of Kechuanning on airway remodeling and the protein level of p-ERK1/2 in lung tissues of asthmatic rats induced by virus. METHODS:The asthmatic rat model induced by respiratory syncytial virus was established. The experimental rats were divided into normal group, asthma model group, low dose (0.33 mL/kg), middle dose (3.0 mL/kg) and high dose (10 mL/kg) of Kechuanning groups, and PD98059 (3 mg/kg) group. The airway responsiveness of the rats was measured by animal ventilator. The pathological changes of the lung tissues were observed by HE staining. PAS staining and Masson staining were used to observe goblet epithelial cells metaplasia and airway collagen deposition. The expression of matrix metalloproteinases-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in the lung tissues of the rats was detected by immunohistochemical staining. The protein levels of ERK1/2 and p-ERK1/2 were determined by Western blot. RESULTS:Compared with model group, the airway responsiveness of the rats in middle dose and high dose of Kechuanning groups was significantly decreased (P<0.01), the injury of lung tissues was significantly decreased, the goblet epithelial cells metaplasia and airway collagen deposition were significantly reduced (P<0.01), and the expression of MMP-9 and TIMP-1 in the lung tissues was also significantly decreased (P<0.01). In addition, the protein level of p-ERK1/2 in high dose of Kechuanning group was significantly decreased compared with model group (P<0.01). CONCLUSION:Kechuanning may treat asthma by regulating the expression of p-ERK1/2 in the lung tissues and improving the airway remodeling symptoms of asthmatic rats induced by virus.     

Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens

We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control.     

基于nanoUPLC-MS/MS的螺旋藻游离肽组成分析

为了研究螺旋藻中的多肽成分,以钝顶螺旋藻为原料,采用超滤法提取螺旋藻中的游离肽,利用nanoUPLC-MS/MS和PEAKS Studio分析谱图信息,结合NCBI数据库进行比对和从头测序分析,获得游离肽的结构组成和百分含量信息。结果表明,利用数据库比对法和从头测序法,分别得到可信的游离肽4 485个和20 597个,匹配到的蛋白质有1 036种。数据库比对结果中的游离肽主要为七肽到二十一肽,少量为二十一肽以上;从头测序结果中游离肽主要为二肽到十六肽,少量为十六肽以上。数据库比对结果中百分含量最高的游离肽是十肽,为15.95%;从头测序结果中百分含量最高的是五肽,达到24.09%。本研究结果为螺旋藻蛋白资源的进一步开发和利用提供了依据。     

超高效液相色谱-串联质谱法测定牛奶和奶粉中五种苯并咪唑类药物

为建立牛奶及奶粉中5种苯并咪唑类药物(阿苯达唑、芬苯达唑、奥芬达唑、噻苯达唑、苯硫氨酯)简便、准确的分析检测方法,以固相萃取为净化手段,采用Acquity UPLC^® BEH C₁₈(100 mm×2.1 mm,1.7 μm)色谱柱进行分离,电喷雾离子源(ESI⁺),多反应监测(MRM)模式检测。结果表明,5种苯并咪唑类药物在1~500 μg·L^-1范围内呈线性相关,相关系数(r)均大于0.996,加标平均回收率介于91.7%~97.8%之间,相对标准偏差(RSD)在1.4%~6.5%之间,基质效应均小于15%。5种药物的检出限均在0.1~3 μg·kg^-1范围内,定量限均在0.5~10 μg·kg^-1范围内。该方法操作简单、快速且灵敏度高,重现性好,能够用于牛奶及奶粉5种苯并咪唑类药物的同时测定。     

有机质浸染砂水泥土的力学特性及本构关系

为研究海南省海湾地区分布的有机质浸染砂水泥土的力学特性,该文首先对有机质浸染砂水泥土和标准砂水泥土进行无侧限抗压强度对比试验(采用配合比均为熟石灰掺入比7.5%、水泥掺入比20%、水灰比0.45),定量分析养护龄期对其无侧限抗压强度及试样破坏形式的影响。然后对有机质浸染砂水泥土进行单轴抗压试验,获得了水泥土材料的应力-应变全过程曲线、刚度变化规律以及改进邓肯-张本构模型。结果表明:1)有机质浸染砂水泥土试样的破坏类型为塑性剪切破坏和脆性剪切破坏;2)有机质浸染砂水泥土抗压强度随着养护龄期的增加基本呈指数形式增长,但在养护龄期14 d后,增长速度逐渐降低并趋于稳定;3)随着养护龄期增长,水泥土刚度增加。在加载初期,水泥土切线模量随着轴向应变增加而增大,呈现刚度硬化现象;4)基于单轴抗压试验得到应力-应变全过程曲线,可分为2个阶段:塑性阶段、软化阶段;5)通过对应力-应变全曲线的描述,得到了修正的邓肯-张模型,确定模型参数后,与实测数据对比发现,该修正模型可以模拟有机质浸染砂水泥土的应力-应变关系。     

CeO2添加比例对Fe基催化剂催化纤维素气化制氢的影响

为研究CeO2添加对生物质催化气化制氢特性的影响，该研究采用分级气化系统分析了不同CeO2/Fe2O3比例（Ce∶Fe摩尔比为0∶1、3∶7、5∶5、7∶3、1∶0）双金属催化剂对纤维素水蒸气催化重整制氢气体产物产量、组成以及催化剂的结构演变特性的影响。结果表明，CeO2/Fe2O3催化剂在制氢反应中的催化性能明显优于纯CeO2或Fe2O3催化剂，当Ce∶Fe摩尔比为3∶7时，在800℃下氢气的最大产率为21.63 mmol/g（以纤维素计，下同）；当温度大于等于800℃时，催化剂氧化还原反应后可生成CeFeO3，且CeFeO3的存在对纤维素水蒸气气化过程有促进作用。CeO2的引入提高了催化剂的氧化性能和稳定性，提高了使用寿命。该研究对生物质气化机制的深入理解具有一定的指导意义。     

东北黑土区坡耕地斜坡垄作与顺坡垄作土壤侵蚀的对比分析

斜坡垄作是东北黑土区最普遍的垄作方式之一，但当前关于斜坡垄作对坡耕地土壤侵蚀的影响鲜见报道。为此，本研究基于室内模拟试验，设计2个降雨强度（50和100 mm/h）以及2种垄作方式（斜坡垄作和顺坡垄作），分析东北黑土区坡耕地斜坡垄作与顺坡垄作坡面土壤侵蚀的差异。结果表明：（1）在50和100 mm/h降雨强度下，斜坡垄作断垄前坡面侵蚀速率分别是顺坡垄作的0.46%和0.35%；但在45 min的降雨过程中，由于斜坡垄作发生断垄现象，造成50和100 mm/h两种降雨强度下斜坡垄作坡面侵蚀速率分别是顺坡垄作的1.24和1.03倍。（2）斜坡垄作径流强度和侵蚀速率随降雨历时的变化均从断垄开始发生突变。在50和100 mm/h降雨强度下，随着降雨历时的变化斜坡垄作断垄前的径流强度和侵蚀速率值均低于顺坡垄作,其平均径流强度分别为顺坡垄作的8.42%和3.75%、平均侵蚀速率分别为顺坡垄作的0.46%和0.35%，但斜坡垄作断垄后坡面径流和侵蚀速率明显增大，其平均径流强度分别为顺坡垄作的1.33和1.47倍、平均侵蚀速率分别是顺坡垄作的2.03和1.62倍。（3）在50和100 mm/h降雨强度下，斜坡垄作断垄前坡面径流量和侵蚀量存在显著的线性关系（P<0.01），而断垄后两者的相关关系则不显著（P>0.05）；而顺坡垄作在两种降雨强度下坡面径流量和侵蚀量存在显著的线性关系。（4）在两种降雨强度下斜坡垄作坡面90%以上的径流泥沙均来自断垄后。因此，提高垄丘稳定性和防止断垄现象发生，是减少斜坡垄作坡面土壤侵蚀的关键所在。     

CUDC-907 induces DNA damage and autophagy in glioma cells

AIM:To investigate the effect of CUDC-907, a dual histone deacetylase (HDAC) and phosphatidylinositol 3-kinase (PI3K) inhibitor, on the DNA damage, cell cycle distribution and autophagy in human glioma U251 cells. METHODS:U251 cells were treated with CUDC-907 of different concentrations, and the cell viability was detected by MTT assay. The quantitative γ-H2AX foci were determined by laser scanning confocal microscopy. The cell cycle distribution of U251 cells was examined by flow cytometry. The protein expression was determined by Western blot analysis. RESULTS:CUDC-907 inhibited the cell viability and the phosphorylation of Akt and p70 ribosomal protein S6 kinase (p70s6K) in the U251 cells (P<0.05). In CUDC-907-treated cells, the number of γ-H2AX foci and protein expression of γ-H2AX were increased significantly (P<0.05). CUDC-907 also induced cell arrest in the G₂/M phase by up-regulating the expression of p21, and inhibiting the protein level of cyclin B1 and the phosphorylation of cell division cycle protein 2 (Cdc2). In addition, CUDC-907 triggered cell autophagy, and inhibition of autophagy increased CUDC-907-induced DNA damage of U251 cells. CONCLUSION:CUDC-907 significantly inhibits PI3K/Akt signaling pathway, induces DNA damage and arrests cell cycle in G₂/M phase. Blockage of autophagy promotes CUDC-907-induced DNA damage of U251 cells.     

基于UPLC-QTOF/MS的小麦发芽代谢组学分析方法

为探究不同提取方法对发芽小麦代谢物提取效果的影响,本研究建立了基于超高效液相色谱-四级杆飞行时间质谱(UPLC-QTOF/MS)的发芽小麦非靶向代谢组学样品前处理方法和分析方法,以发芽2 d周麦26籽粒为材料,设计提取溶剂、提取方式、提取时间3个因素在3个水平上L9(34)正交试验,并通过主成分分析和聚类分析确定提取效果最佳的组合。以80%乙腈(0.1%甲酸)震摇提取30 min可检测出1609个代谢峰,说明提取溶剂对代谢物提取效果起主要作用。共鉴定出92种小麦代谢物。