Effect of Psammaplin A on in vitro Development of Bovine Aging Oocytes

To investigate the effect of histone deacetylation inhibitor Psammaplin A (PsA) on the development of bovine aging oocytes in vitro,oocytes were randomly divided into control group,aging group and 50 mmol/L PsA treated aging group (PsA group).Immunofluorescence staining and JC-1 were used to detect the blastocyst rate of bovine oocytes after parthenogenetic activation,the number of cells in blastocysts,apoptosis,reactive oxygen species (ROS),glutathione (GSH) and mitochondrial membrane potential intensity of embryos.The results showed that the blastocyst rate of the aging group was significantly lower than that of PsA and control groups (P<0.05).The blastocyst rate of PsA group was not significantly different from that of control group (P>0.05).The number of cells in the blastocysts of control group and PsA group were significantly higher than that of aging group (P<0.05).The number of cells in the blastocysts of PsA group was not significantly different from that of control group (P>0.05).The apoptosis rate in aging group was significantly higher than that of control and PsA groups (P<0.05),the apoptosis rate of PsA group was significantly higher than that of control group (P<0.05).The GSH level of MⅡ oocytes in aging group was significantly lower than that of control and PsA groups (P<0.05).There was no significant difference in GSH level between control and PsA groups (P>0.05).The ROS level of the embryos in aging group was significantly higher than that of control and PsA groups (P<0.05).The ROS level in PsA group was significantly higher than that of control group (P<0.05).The mitochondrial membrane potential of early embryos of aging group 4-8 cells was significantly lower than that of control and PsA groups (P<0.05).The mitochondrial membrane potential intensity of control group was significantly higher than that of PsA group (P<0.05).In summary,PsA could effectively delay the aging of bovine oocytes and improve the quality of oocytes.     

Evaluating biodegradability of soil organic matter by its thermal stability and chemical composition

The stability of soil organic matter (SOM) as it relates to resistance to microbial degradation has important implications for nutrient cycling, emission of greenhouse gases, and C sequestration. Hence, there is interest in developing new ways to quantify and characterise the labile and stable forms of SOM. Our objective in this study was to evaluate SOM under widely contrasting management regimes to determine whether the variation in chemical composition and resistance to pyrolysis observed for various constituent C fractions could be related to their resistance to decomposition. Samples from the same soil under permanent pasture, an arable cropping rotation, and chemical fallow were physically fractionated (sand: 2000-50 μm; silt: 50-5 μm, and clay: <5 μm). Biodegradability of the SOM in size fractions and whole soils was assessed in a laboratory mineralization study. Thermal stability was determined by analytical pyrolysis using a Rock-Eval pyrolyser, and chemical composition was characterized by X-ray absorption near-edge structure (XANES) spectroscopy at the C and N K-edges. Relative to the pasture soil, SOM in the arable and fallow soils declined by 30% and 40%, respectively. The mineralization bioassay showed that SOM in whole soil and soil fractions under fallow was less susceptible to biodegradation than that in other management practices. The SOM in the sand fraction was significantly more biodegradable than that in the silt or clay fractions. Analysis by XANES showed a proportional increase in carboxylates and a reduction in amides (protein) and aromatics in the fallow whole soil compared to the pasture and arable soils. Moreover, protein depletion was greatest in the sand fraction of the fallow soil. Sand fractions in fallow and arable soils were, however, relatively enriched in plant-derived phenols, aromatics, and carboxylates compared to the sand fraction of pasture soils. Analytical pyrolysis showed distinct differences in the thermal stability of SOM among the whole soil and their size fractions; it also showed that the loss of SOM generally involved preferential degradation of H-rich compounds. The temperature at which half of the C was pyrolyzed was strongly correlated with mineralizable C, providing good evidence for a link between the biological and thermal stability of SOM.     

类人机器人全能运动的稳定性研究

类人机器人全能运动包括不同粗糙平面上的快速行走、越障、踢球、格斗等,稳定性是上述运动的关键环节。本文从分析机器人关节角与零力矩点（ZMP）的关系入手,研究提高机器人运动稳定性的新方法。具体步骤是先粗调校正机器人姿态,然后确定各关节角度,得到保证稳定运动的关节角范围。最后依托韩国GP型机器人进行测试,结果表明了所提出方法的有效性。     

重组猪抑制素蛋白质中毒素残留检测及其理化稳定性

为探索纯化的重组猪抑制素蛋白质毒素残留情况以及重组猪抑制素蛋白质稳定性,本研究利用鲎试剂、气相色谱和SDS-PAGE方法对重组猪抑制素蛋白质中的细菌内毒素、β-巯基乙醇的残留量及其蛋白质在不同温度、极端p H值和反复冻融条件下的稳定性进行检测。结果显示,经过3次等电点沉淀洗涤后,重组猪抑制素蛋白质中细菌内毒素和β-巯基乙醇的残留量大幅减少。通过在不同温度下的处理发现,重组猪抑制素蛋白质在4℃环境下较稳定,可以长期保存;但在37℃时,稳定性快速下降,随着温度上升而降解加速。通过极端p H值和反复冻融试验发现,重组猪抑制素蛋白质对极端p H值和反复冻融具有较好的耐受性。     

Strong functional stability of soil microbial communities under semiarid Mediterranean conditions and subjected to long-term shifts in baseline precipitation

We investigated the effect of soil microclimate on the structure and functioning of soil microbial communities in a Mediterranean Holm-oak forest subjected to 10 years of partial rain exclusion manipulations, simulating average drought conditions expected in Mediterranean areas for the following decades. We applied a high throughput DNA pyrosequencing technique coupled to parallel measurements of microbial respiration (R_H) and temperature sensitivity of microbial respiration (Q₁₀). Some consistent changes in the structure of bacterial communities suggest a slow process of community shifts parallel to the trend towards oligotrophy in response to long-term droughts. However, the structure of bacterial communities was mainly determined by short-term environmental fluctuations associated with sampling date (winter, spring and summer) rather than long-term (10 years) shifts in baseline precipitation. Moreover, long-term drought did not exert any chronic effect on the functioning of soil microbial communities (R_H and Q₁₀), emphasizing the functional stability of these communities to this long-term but mild shifts in water availability. We hypothesize that the particular conditions of the Mediterranean climate with strong seasonal shifts in both temperature and soil water availability but also characterized by very extreme environmental conditions during summer, was acting as a strong force in community assembling, selecting phenotypes adapted to the semiarid conditions characterizing Mediterranean ecosystems. Relations of climate with the phylogenetic structure and overall diversity of the communities as well as the distribution of the individual responses of different lineages (genera) to climate confirmed our hypotheses, evidencing communities dominated by thermotolerant and drought-tolerant phenotypes.     

基于状态变换法的车辆悬架系统时滞反馈控制

为了提高车辆行驶平顺性和稳定性,研究悬架系统中时滞补偿控制效果,本文以赛欧轿车悬架系统为基础,建立2自由度车辆半主动悬架系统模型,设计了时滞反馈控制器,采用理论与试验相结合的方法对系统时滞反馈控制特性进行研究。首先建立含时滞的悬架系统动力学方程,采用常微分理论和多项式判别方法分析系统稳定性,并通过时域与频域仿真对结果进行验证。研究表明:采用传统二次型最优控制律对含时滞的悬架系统进行控制,当系统控制时滞较大时,系统定性特性可能会发生改变,甚至会失稳发散。为保证系统的稳定性,采用状态变换方法设计时滞反馈最优控制律,仿真表明采用该控制律不仅可以保证系统稳定性,系统的减振特性亦有改善。最后搭建了悬架时滞反馈控制平台,基于时域辨识方法测得系统固有时滞为0.065 s,通过对相同工况下仿真结果与试验结果进行对比,发现两者具有较好的一致性,误差在15%以内,满足业内使用要求,表明研究可信,结果可为主动/半主动车辆悬架控制器实际设计应用提供参考。     

马链球菌兽疫亚种烯醇化酶对小鼠肺泡巨噬细胞吞噬功能的影响

旨在研究马链球菌兽疫亚种（Streptococcus equi ssp.zooepidemicus，SEZ）烯醇化酶（enolase，Eno）对小鼠肺泡巨噬细胞（RAW264.7）吞噬能力的影响。通过构建原核表达质粒获得重组烯醇化酶（rEno），采用台盼蓝活细胞计数法，判定在不同处理浓度和时间下rEno蛋白对RAW264.7细胞的细胞毒性。将rEno蛋白与RAW264.7细胞共孵育后，用SEZ作用于细胞并检测细胞吞菌数量，判断RAW264.7细胞对SEZ的吞噬活性。进一步通过活细胞稳定同位素标记技术（SILAC）和蛋白质谱分析技术（LC-MS/MS），筛选到RAW264.7细胞中可能与SEZ Eno存在相互作用的候选蛋白。结果发现，10 μg·mL^-1 rEno蛋白处理对RAW264.7细胞有明显的细胞毒性，且10 μg·mL^-1 rEno蛋白处理RAW264.7细胞2和4 h可显著抑制其对SEZ的吞噬作用（P<0.01、P<0.05）。初步筛选到RAW264.7细胞中动力蛋白激活蛋白亚单位蛋白（dynactin subunit protein 2，Dctn）、整合素α-M蛋白（integrin alpha-M）等17种可能与Eno发生互作的蛋白。本研究获得了rEno重组表达蛋白，发现rEno可减少RAW264.7细胞对SEZ的吞噬，互作蛋白的初步筛选也为进一步揭示Eno在SEZ抗吞噬中的作用机制奠定了基础。     

Vitamin C enhanced myocardial differentiation of dedifferentiated fat cells

AIM: In order to observe the myocardial differentiation capacity of the dedifferentiated fat (DFAT) cells treated with vitamin C in vitro. METHODS: DFAT cells were dedifferentiated from the mature rat adipocytes with ceiling adherent culture. The DFAT cells of passage 3 were used in the study. Vitamin C and/or neonatal rat heart tissue lysate were added into the culture medium to induce myocardial differentiation for 3 weeks. The cell morphology was observed under microscope. The myocardial-specific markers, such as cTnT, GATA-4 and NKx2.5, were examined by the methods of immunofluorescence, PCR and Western blot. RESULTS: Mature rat adipocytes dedifferentiated into fibroblast-like DFAT cells after ceiling adherent culture. The DFAT cells spontaneously differentiated into cardiomyocyte-like cells under normal culture condition with a low incidence. After treated with neonatal rat heart cell lysate, the DFAT cells became cardiomyocyte-like cells that had bigger size, longer shape and myotubule-structure. The expression of cTnT, GATA-4 and NKx2.5 was remarkably increased at both mRNA and protein levels as compared with the normal cultured DFAT cells. The expression of cTnT, GATA-4 and NKx2.5 was further increased in DFAT cells after treating with vitamin C. No spontaneous beating cell was observed. CONCLUSION: Vitamin C enhances the differentiation of DFAT cells into cardiomyocyte-like cells.     

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells

Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.