EBNA3C与Gemin3两种蛋白形成复合物及相互作用的结构域

探讨EB病毒核抗原3C(EBNA3C)与Gemin3的相互结合及其作用区域。用Flag-EBAN3C与GFPGemin3共转染HEK 293细胞,采用免疫共沉淀、谷胱苷肽-S转移酶共沉淀及免疫荧光蛋白共存实验确定EBNA3C与Gemin3两种蛋白在体内、体外相互结合及相互作用的结构域。EBNA3C与Gemin3两种蛋白在体内外相结合,二者通过各自的C端相互结合。EBNA3C与Gemin3两种蛋白形成稳定复合物。     

Molecular insights into the binding selectivity of a synthetic ligand DAAG to Fc fragment of IgG

Affinity chromatography with synthetic ligands has been focused as the potential alternative to protein A‐based chromatography for antibody capture because of its comparable selectivity and efficiency. Better understanding on the molecular interactions between synthetic ligand and antibody is crucial for improving and designing novel ligands. In this work, the molecular interaction mechanism between Fc fragment of IgG and a synthetic ligand (DAAG) was studied with molecular docking and dynamics simulation. The docking results on the consensus binding site (CBS) indicated that DAAG could bind to the CBS with the favorable orientation like a tripod for the top‐ranked binding complexes. The ligand‐Fc fragment complexes were then tested by molecular dynamics simulation at neutral condition (pH 7.0) for 10 ns. The results indicated that the binding of DAAG on the CBS of Fc fragment was achieved by the multimodal interactions, combining the hydrophobic interaction, electrostatic interaction, hydrogen bond, and so on. It was also found that multiple secondary interactions endowed DAAG with an excellent selectivity to Fc fragment. In addition, molecular dynamics simulation conducted at acidic condition (pH 3.0) showed that the departure of DAAG ligand from the surface of Fc fragment was the result of reduced interaction energies. The binding modes between DAAG and CBS not only shed light on the molecular mechanisms of DAAG for antibody purification but also provide useful information for the improvement of ligand design. Copyright © 2014 John Wiley & Sons, Ltd.     

Precise chronology of differentiation of developing human primary dentition

While correlation of developmental stage with embryonic age of the human primary dentition has been well documented, the available information regarding the differentiation timing of the primary teeth was largely based on the observation of initial mineralization and varies significantly. In this study, we aimed to document precise differentiation timing of the developing human primary dentition. We systematically examined the expression of odontogenic differentiation markers along with the formation of mineralized tissue in each developing maxillary and mandibular teeth from human embryos with well-defined embryonic age. We show that, despite that all primary teeth initiate development at the same time, odontogenic differentiation begins in the maxillary incisors at the 15th week and in the mandibular incisors at the 16th week of gestation, followed by the canine, the first primary premolar, and the second primary premolar at a week interval sequentially. Despite that the mandibular primary incisors erupt earlier than the maxillary incisors, this distal to proximal sequential differentiation of the human primary dentition coincides in general with the sequence of tooth eruption. Our results provide an accurate chronology of odontogenic differentiation of the developing human primary dentition, which could be used as reference for future studies of human tooth development.     

The miR156‐SPL9‐DFR pathway coordinates the relationship between development and abiotic stress tolerance in plants

Young organisms have relatively strong resistance to diseases and adverse conditions. When confronted with adversity, the process of development is delayed in plants. This phenomenon is thought to result from the rebalancing of energy, which helps plants to coordinate the relationship between development and stress tolerance; however, the molecular mechanism underlying this phenomenon remains mysterious. In this study, we found that miR156 integrates environmental signals to ensure timely flowering, thus enabling the completion of breeding. Under stress conditions, miR156 is induced to maintain the plant in the juvenile state for a relatively long period of time, whereas under favorable conditions, miR156 is suppressed to accelerate the developmental transition. Blocking the miR156 signaling pathway in Arabidopsis thaliana with 35S::MIM156 (via target mimicry) increased the sensitivity of the plant to stress treatment, whereas overexpression of miR156 increased stress tolerance. In fact, this mechanism is also conserved in Oryza sativa (rice). We also identified downstream genes of miR156, i.e. SQUAMOSA PROMOTER BINDING PROTEIN‐LIKE 9 (SPL9) and DIHYDROFLAVONOL‐4‐REDUCTASE (DFR), which take part in this process by influencing the metabolism of anthocyanin. Our results uncover a molecular mechanism for plant adaptation to the environment through the miR156‐SPLs‐DFR pathway, which coordinates development and abiotic stress tolerance.     

Isolation and passage of muscle-derived stem cells from the rat penile corpora cavernosa and induction of differentiation into smooth muscle cells

This study treated the isolation and passage of muscle-derived stem cells (MDSCs) from rat penile corpora cavernosa, detection of stem cell marker expression, observation of their self-renewal and continuous proliferation, and demonstration of their potential to differentiate into smooth muscle cells in co-culture. Muscle-derived stem cells from the rat penile corpora cavernosa were isolated and purified. The expression of stem cell markers Sca-1 and desmin was detected in PP6 cells, thus confirming that the main components of PP6 cells are MDSCs. The expression of Sca-1 and desmin occurred both in PP6 cells and cells at passages 3, 6, and 8, and there was no significant decrease in the expression level with increasing passage number. The growth curves indicated that the cell doubling time was approximately 48 h. The cells entered the stationary phase after approximately 7 days of culture. The proliferative activity of the cells at passage 8 remained unchanged. After 2 days of co-culture with smooth muscle cells, the DAPI-labeled MDSCs tended to exhibit smooth muscle cell morphology and expression of α-SMA was detected. MDSCs exist in the rat penile corpora cavernosa and possess the potential to differentiate into smooth muscle cells. This discovery serves as the basis in view of the potential use of endogenous stem cells for the treatment of erectile dysfunction (ED).     

不同互花米草治理措施对植物与大型底栖动物的影响简

治理外来入侵植物互花米草(Spartina alterniflora)对保护河口湿地鸟类栖息地及生物多样性具有重要意义。研究在崇明东滩比较了"淹水刈割"、"反复刈割"及"化学除草"三种措施对大面积互花米草的治理效果及其对大型底栖动物与土著植物芦苇(Phragmites australis)的影响。结果表明,反复刈割措施对互花米草生长具有一定的控制作用,对底栖动物群落的影响较小;使用化学除草剂清除互花米草的效果不明显,对底栖动物群落的影响亦不明显;淹水刈割措施能长期有效地清除互花米草,但长期淹水对底栖动物群落的影响较大,同时亦对芦苇生长造成一定负面影响。因此,淹水刈割可能是在河口生态系统治理大面积互花米草最有效的方法,但是在后续管理中需要采取一定的措施来减小对底栖动物及土著植物的影响。     

氮磷钾配比施肥对广金钱草产量及质量的影响

测定了不同氮磷钾配施条件下广金钱草的株高、地径、种子产量、药材干重及夏佛塔苷含量。结果表明:施肥组生长状况及产量与不施肥组具有极显著差异,其中单施钾肥30g·m-2的广金钱草株高及药材干重最高,分别为174.04cm、94.50g;单施氮肥40g·m-2地径最粗,达到16.79mm;施氮、磷肥各80g·m-2,钾肥30g·m-2种子产量最高,增产率达到179.30%;施氮肥40g·m-2,磷肥80g·m-2,钾肥80g·m-2夏佛塔苷含量最高,达到0.175%。由此可知氮磷钾的合理配施能够促进广金钱草的生长并提高产量,同时影响夏佛塔苷的积累。以株高、地径、种子产量、药材干重、夏佛塔苷含量为评价标准。根据氮磷钾肥料多元二次曲线回归方程数学模型计算结果,得出氮最佳施肥量:42.4～64.5g·m-2,磷:49.5～59.0g·m-2,钾:30.0～41.0g·m-2。氮磷钾最佳施肥组合为N∶P∶K=5.4∶5.5∶3.7。