构建基于CRISPR/Cas9技术敲除ALOX5的质粒

旨在利用CRISPR/Cas9技术构建敲除花生四烯5-脂氧合酶基因(Arachidonate 5-lipoxygenase gene,ALOX5)的重组质粒。设计合成3对靶向敲除ALOX5第六外显子的sgRNA,将其分别插入到CRISPR/Cas9质粒骨架pX458载体中,转化感受态大肠杆菌DH5α后挑取克隆,通过测序评估重组质粒是否构建成功。将构建好的重组质粒转染293T细胞,在荧光显微镜下观察转染效果,挑取转染成功的细胞,用试剂盒提取转染细胞基因组DNA,PCR扩增含敲除位点的DNA片段,用测序技术获得核苷酸序列,用DNAStar软件分析转染细胞中ALOX5基因敲除情况。测序结果表明2对双链sgRNA寡核苷酸已插入质粒,且序列正确,靶向ALOX5基因的重组质粒pX458-sgRNAs-ALOX5构建成功。其在293T细胞中的转染效率约为50%,用一代测序法未检测到sgRNAs的切割效果。初步表明利用CRISPR/Cas9技术成功构建靶向ALOX5基因的重组质粒pX458-sgRNAs-ALOX5。     

中国三种常见蒿属植物潜在地理分布及其主导气候因子

裂叶蒿(Artemisia tanacetifolia)、大籽蒿(Artemisia sieversiana)和艾(Artemisia argyi)是我国常见的蒿属(Artemisia)植物,其分布区域遍布全国。本文利用MaxEnt模型预测3种蒿属植物在当前气候条件以及未来两种气候情景下的潜在分布区。采用受试者工作特征曲线(receiver operating characteristic curve,ROC)检验模型精度。训练数据和测试数据的AUC值均在0.8以上,表明预测结果可靠性良好。在当前的气候条件下,裂叶蒿最适分布区主要为黄土高原、内蒙古高原和东北平原;大籽蒿的最适分布区为西藏南部谷地、横断山地区、黄土高原、内蒙古高原和东北平原;艾的最适分布区有两个,一个位于台湾岛南部,另一个为大巴山、巫山、云贵高原北部、黄土高原和东北平原南部区域。2070年RCP2.6和RCP8.5情景下,裂叶蒿及大籽蒿的高适宜区面积减小,艾的最适分布区面积增加。Jackknife检验结果表明,年均降水量是预测裂叶蒿分布最有效的气候因子,5月降水是预测大籽蒿分布的最显著的气候因子,8月水汽压对艾的影响最大。本研究结果为蒿属植物资源的合理利用提供了科学依据。     

Identification of Human Vital Functions Directly Relevant to the Respiratory System Based on the Cardiac and Acoustic Parameters and Random Forest

Regarding sleep research, polysomnography (PSG) also called a sleep study, is a gold standard. It incorporates brain waves, the oxygen level in the blood, heart rate and breathing, and leg movement recordings. PSG is a complicated and expensive laboratory-based procedure, usually done in hospitals or special sleep center. In this study, an alternative technique for Sleep-Related Breathing Disorders (SRBD) based on selected cardiac and acoustic parameters and the Random Forest (RF) has been studied. A system dedicated to the detection of simultaneously acquired ECG and acoustic signals, which are collected during sleep at home environment is proposed. Results obtained indicate that classification and regression tree models such as RF are appropriate for the evaluation of sleep disorders like SRBD. The best identification of sleep irregularities at level 89.00 percent for the raw database was obtained. Thus, statistical predictive models allow identification of breathing events with high levels of sensitivity and specificity, providing an inexpensive and accurate diagnosis.     

The early life history of two sympatric New Zealand octopuses: eggs and paralarvae of Octopus huttoni and Pinnoctopus cordiformis

This study combined morphological and morphometric information on egg clutches, egg capsules and paralarvae of two sympatric coastal octopuses from New Zealand waters, Octopus huttoni and Pinnoctopus cordiformis, to provide species-specific traits to identify their early life stages obtained from field surveys. Eggs of O. huttoni (2.5 mm length; 1 mm width) were entwined with one another forming strings that ranged from 11 to 25.8 mm in length. Eggs of P. cordiformis (6.4 mm length; 1.5 mm width) were significantly bigger than those of O. huttoni and were grouped in small clusters of about seven eggs. Paralarvae O. huttoni and P. cordiformis differed in hatching size (1.4 mm versus 3.1 mm mantle length), number of suckers per arm (four versus eight), number of lamellae per outer demibranch (five versus ten) and arrangements of chromatophores in the body surface (29 to 59 versus 91 to 179), respectively. The morphological traits described in hatchlings from the laboratory allowed comparisons with field-collected paralarvae, suggesting that such characters were reliable species-specific patterns to enable a consistent differentiation between the early life stages of these two sympatric species, even in the absence of the brooding female.     

长爪沙鼠肝细胞体外分离培养体系的建立

目的建立长爪沙鼠原代肝细胞分离培养体系。方法以雄性长爪沙鼠为供体,采用组织消化法和Seglen两步灌流法分离肝细胞,以台盼蓝染色检测细胞得率和活率,过碘酸-希夫氏反应(PAS)鉴定肝细胞,倒置显微镜观察肝细胞形态变化,并使用含有多种细胞因子的培养基维持培养。结果组织消化法和Seglen两步灌流法平均每只长爪沙鼠可分别获得肝细胞(1.33±0.34)×107个、(3.97±1.15)×107个,细胞活率分别为(29.4±6.05)%、(80.3±4.56)%,这两种方法在细胞得率及活率方面存在显著差异。肝细胞内因有大量的糖原颗粒,经PAS染色后被染成红色。结果表明肝细胞在贴壁后72 h内,肝细胞形态发生显著变化。结论采用胶原酶经肝门静脉灌流分离肝细胞是一种高效获得肝细胞的方法。各种细胞因子有利于维持肝细胞在体外的生长分化,长爪沙鼠原代肝细胞分离培养体系的建立将为肝脏相关疾病研究和防治药物的开发提供技术支持。     

Production of endothelial progenitor cells obtained from human Wharton's jelly using different culture conditions

Endothelial progenitor cells (EPC) participate in revascularization and angiogenesis. EPC can be cultured in vitro from mononuclear cells of peripheral blood, umbilical cord blood or bone marrow; they also can be transdifferentiated from mesenchymal stem cells (MSC). We isolated EPCs from Wharton's jelly (WJ) using two methods. The first method was by obtaining MSC from WJ and characterizing them by flow cytometry and their adipogenic and osteogenic differentiation, then applying endothelial growth differentiating media. The second method was by direct culture of cells derived from WJ into endothelial differentiating media. EPCs were characterized by morphology, Dil-LDL uptake/UEA-1 immunostaining and testing the expression of endothelial markers by flow cytometry and RT-PCR. We found that MSC derived from WJ differentiated into endothelial-like cells using simple culture conditions with endothelium induction agents in the medium.     

Transcriptional profiling identifies critical steps of cell cycle reprogramming necessary for Plasmodiophora brassicae‐driven gall formation in Arabidopsis

Plasmodiophora brassicae is a soil‐borne biotroph whose life cycle involves reprogramming host developmental processes leading to the formation of galls on its underground parts. Formation of such structures involves modification of the host cell cycle leading initially to hyperplasia, increasing the number of cells to be invaded, followed by overgrowth of cells colonised by the pathogen. Here we show that P. brassicae infection stimulates formation of the E2Fa/RBR1 complex and upregulation of MYB3R1, MYB3R4 and A‐ and B‐type cyclin expression. These factors were previously described as important regulators of the G2?M cell cycle checkpoint. As a consequence of this manipulation, a large population of host hypocotyl cells are delayed in cell cycle exit and maintained in the proliferative state. We also report that, during further maturation of galls, enlargement of host cells invaded by the pathogen involves endoreduplication leading to increased ploidy levels. This study characterises two aspects of the cell cycle reprogramming efforts of P. brassicae: systemic, related to the disturbance of host hypocotyl developmental programs by preventing cell cycle exit; and local, related to the stimulation of cell enlargement via increased endocycle activity.     

Effectiveness of 2-Hydroxybenzoic Acid in Physiologic Response of Basket Willow (Salix viminalis L.) on Stress Induced by Cadmium

Russian Journal of Plant Physiology - The aim of the study was an evaluation of the effect of exogenous application of 2-hydroxybenzoic acid on the performance of photosynthetic apparatus of two...     

Designed and naturalised sward response to management: I. Patterns of herbage production

Pastures in the Appalachian region of the United States comprise a mix of grasses, legumes and forbs that tend to differ in productivity within and among years. A high degree of spatial variability in hill‐land pasture creates microsite conditions that influence botanical composition of pasture. The variation in sward composition presents logistical challenges to livestock producers who rely on a dependable supply of herbage mass and nutritive value to meet production goals. Our objective was to determine if forage communities sown for specific functions, for example, superior dry matter productivity, resource patch exploitation or targeted seasonal production, adapted to changing growing conditions within and among years. Productivity of communities differed among years reflecting the cumulative influences of time, ontogenetic and environmental variations. Maximum productivity was influenced by the specific forage community and less so by simple clipping and fertiliser management. Naturalised swards clipped to emulate hay management tended to have sustained herbage productivity but lower nutritive value when compared to sown communities. Rankings of dry matter productivity of communities were similar for each year where bioactive composition, high productivity and warm season tended to produce the most, and stoloniferous‐rhizomatous and naturalised pasture the least. Regardless of initial sward composition, effective number of species as an index of diversity increased when frequently clipped swards were not fertilised, and when infrequently clipped swards were fertilised. Dry matter production patterns were not influenced by the effective number of species in any forage community suggesting that key species sustained productivity with volunteer species making lesser contribution to total productivity. The species composition of forage plant communities appears to be more important than clipping or fertiliser management practices as a means to sustain forage productivity.