鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

超声提取-离子色谱法测定芹菜中氯量

研究了超声提取-离子色谱法测定芹菜中氯含量的方法。芹菜经超声提取20 min后,以浓度4.5 mmol/L Na₂CO₃和0.8 mmol/L NaHCO₃为淋洗液,经IonPacAG23分离测定。本方法具有快速、灵敏、准确度高,适合于芹菜中氯量的测定。     

施氮水平对7S亚基缺失大豆根系形态和结瘤固氮的影响

为有效推广功能型大豆7S亚基缺失品种,以7S亚基缺失大豆品系东富2号为研究对象,设置4种施氮水平(纯N),N0(0 mg·kg~(-1))、N1(25 mg·kg~(-1))、N2(50 mg·kg~(-1))、N3(75 mg·kg~(-1)),采用桶栽法研究大豆根系形态和结瘤固氮对不同施氮水平的响应。结果表明:N1(25 mg·kg~(-1))水平下根系干重加大,根冠比增大,根瘤固氮潜力高,单株产量较高。N2(50 mg·kg~(-1))水平下根长、根表面积、根体积在生育后期增长较快,根系干重较大,根冠比低,固氮酶活性最高,单株籽粒产量最高。N3(75 mg·kg~(-1))水平下植株干重较大,无效生长较多,根瘤数少,固氮潜力和根冠比低,单株产量不高。综合籽粒产量和根系特性指标,功能型大豆7S亚基缺失品系东富2号的适宜施肥量为25~50 mg·kg~(-1)。     

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.     

Zerumbone attenuates MPP+-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.     

Changes in wettability of wood surface using electron beam irradiation

ABSTRACT

This study deals with the influence of electron beam irradiation (EBI) on the wettability of Norway spruce surface samples. To evaluate this possible effect in detail, the changes in chemistry and wood structure were analysed using the methods of IR spectroscopy and Scanning Electron Microscopy (SEM). The surface wettability was determined by the contact angle measurement method. The results of infrared spectra show decreased relative numbers of available hydrophilic groups (e.g. hydroxyl and carbonyl groups) in the wood surfaces due to the electron beam irradiation. This behaviour is consistent with the values from the surface wettability measurements. Structural differences of the wood surfaces after electron beam irradiation were not detectable. Based on these findings pre-treatment of wood surfaces for industrial usages can be a potential field of application of this technology.     

干旱胁迫下不同品种杨树矿质离子含量变化

【目的】了解杨树在干旱胁迫下矿质离子分布及其动态变化。【方法】以吴屯杨、新疆杨和荷兰杨为实验材料,在中度和重度干旱条件下测定了根、茎、叶和质外体的K^+、Ca^2+、Na^+和Mg^2+等4种离子含量的变化。【结果】干旱胁迫使K^+、Ca^2+和Na^+离子含量上升,Mg^2+含量下降,其中K^+和Ca^2+含量自胁迫开始明显升高,Na^+含量则在中度胁迫之后明显增加。离子在各器官分布情况为:K^+主要分布于杨树的叶片,Ca^2+、Na^+和Mg^2+主要分布于杨树根部,且离子分布存在品种间差异,荷兰杨和新疆杨中K^+分布为:叶>根>茎,而吴屯杨体内K^+分布则为:叶>茎>根;3种杨树中Ca^2+分布均为:根>叶>茎;吴屯杨和荷兰杨中Na^+分布为:根>茎>叶,新疆杨则为:根>叶>茎;在Mg^2+的分布上吴屯杨和新疆杨较为类似即:根>叶>茎,而荷兰杨为叶>根>茎。【结论】干旱胁迫下杨树首选吸收的离子是K^+和Ca^2+,K^+主要分布在杨树的叶片中,其次是根部,Ca^2+主要分布在杨树的根部。重度干旱胁迫下,杨树开始吸收Na^+,储存在杨树的根部。不同品种杨树之间离子的分布和吸收都存在品种差异。质外体作为离子的运输通道,在各干旱条件下运输4种矿质离子至杨树的其他部位,Mg^2+存在滞留在质外体中的情况。本研究可为植物在干旱胁迫下矿质离子吸收和分布的研究提供理论依据。     

极端暴雨条件下黄土丘陵沟壑区土壤蓄水能力和入渗规律

黄土高原退耕还林还草工程实施后,下垫面环境条件的变化可能对流域水文过程、水文通量、水量平衡以及生态系统产生十分重要的影响。研究极端暴雨条件下剖面土壤蓄水能力和入渗规律,对于阐明流域产汇流过程和影响机制具有重要的科学价值。采用土壤墒情仪对陕北"7·26"特大暴雨事件下黄土丘陵沟壑区草地剖面土壤水分进行了实时动态监测,分析了极端暴雨条件下剖面土壤水分的动态变化和蓄水过程,利用Horton入渗模型模拟了剖面土壤水分湿润锋的运动过程,揭示了极端暴雨条件下剖面土壤水分的入渗规律。结果表明:(1)极端暴雨条件下,黄土丘陵沟壑区坡面草地不同深度层次土壤水分与降雨过程的响应不同,具有层次性和明显的滞后效应,其中,0～140 cm是影响该地区土壤水文过程的关键层次;(2)土壤水分再分配结束时,湿润锋最深深度达140 cm,土壤蓄水量达225.99 mm,较降雨前95.37 mm增加了1.37倍;(3)极端暴雨过程中湿润锋的运动随时间呈对数递减关系,其稳渗速率随容重增加而减小,呈指数函数递减;(4)极端降雨过程中该地区坡面草地的产流机制仍以超渗产流为主,对于揭示流域的产汇流机制和完善水文预报模型具有重要的科学意义。     

预应力薄腹梁质量事故的分析及鉴定

阐述了某工程由于施工原因，导致预应力混凝土工字形屋面梁强度不足的预制构件的检验鉴定过程和方法。采用回弹法推定混凝土实际强度；通过静载试验测试构件的实际挠度、抗裂和承载能力。分析了导致混凝土强度不足的原因。     

T7 RNA多聚酶基因的克隆、序列测定及其在E.coli中的表达

从BL21(DE3)E．coli菌株中以PCR的方法扩增得到了与T7RNA多聚酶(T7RNApolymerase，T7pol)基因大小一致的DNA片断。将PCR产物纯化后直接克隆到pGEM—T载体中，经酶切鉴定和DNA序列分析表明克隆得到了正确的T7pol基因。将T7pol基因亚克隆入pET-28b( )中，构建得到原核表达质粒pET28T7。该质粒的BL21(DE3)pLysS转化菌在IPTG的诱导下可表达约98800的蛋白，这与T7pol的相对分子质量一致。将该质粒转化DH5α、JMl09、HBl01、BL21(DE3)和BL21(DE3)pLysS等5种不同的宿主菌，仅有转化T7pol酶活性受到抑制的宿主菌BL21(DE3)pLysS才能得到转化子，而其余4种T7T7pol酶活性不受抑制的E．coli宿主菌不能得到转化子。pET28T7原核表达质粒这种仅能在T7pol酶活性受到抑制的宿主菌中才能存活的现象说明本试验所克隆的T7pol基因能正确表达出具有RNA转录酶活性的蛋白。