Triptolide Inhibited Cytotoxicity of Differentiated PC12 Cells Induced by Amyloid-Beta25–35 via the Autophagy Pathway

Evidence shows that an abnormal deposition of amyloid beta-peptide_25–35 (Aβ_25–35) was the primary cause of the pathogenesis of Alzheimer’s disease (AD). And the elimination of Aβ_25–35 is considered an important target for the treatment of AD. Triptolide (TP), isolated from Tripterygium wilfordii Hook.f. (TWHF), has been shown to possess a broad spectrum of biological profiles, including neurotrophic and neuroprotective effects. In our study investigating the effect and potential mechanism of triptolide on cytotoxicity of differentiated rat pheochromocytoma cell line (the PC12 cell line is often used as a neuronal developmental model) induced by Amyloid-Beta_25–35 (Aβ_25–35), we used 3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MTT) assay, flow cytometry, Western blot, and acridine orange staining to detect whether triptolide could inhibit Aβ_25–35–induced cell apoptosis. We focused on the potential role of the autophagy pathway in Aβ_25–35-treated differentiated PC12 cells. Our experiments show that cell viability is significantly decreased, and the apoptosis increased in Aβ_25–35-treated differentiated PC12 cells. Meanwhile, Aβ_25–35 treatment increased the expression of microtubule-associated protein light chain 3 II (LC3 II), which indicates an activation of autophagy. However, triptolide could protect differentiated PC12 cells against Aβ_25–35-induced cytotoxicity and attenuate Aβ_25–35-induced differentiated PC12 cell apoptosis. Triptolide could also suppress the level of autophagy. In order to assess the effect of autophagy on the protective effects of triptolide in differentiated PC12 cells treated with Aβ_25–35, we used 3-Methyladenine (3-MA, an autophagy inhibitor) and rapamycin (an autophagy activator). MTT assay showed that 3-MA elevated cell viability compared with the Aβ_25–35-treated group and rapamycin inhibits the protection of triptolide. These results suggest that triptolide will repair the neurological damage in AD caused by deposition of Aβ_25–35 via the autophagy pathway, all of which may provide an exciting view of the potential application of triptolide or TWHF as a future research for AD.     

垂直绿化生态效益研究综述

全球气候变化和城市化背景下城市生态环境问题越发凸显，面对日趋紧张的土地资源现状，垂直绿化建设成为重要的城市绿地补充手段，对改善城市环境具有重要意义。垂直绿化具有多种生态效益，且其效益发挥受到多因素共同影响。如何统筹协调生态效益和影响因素之间的复杂关系，以使其综合生态效益最大化，是当前垂直绿化建设亟待解决的科学问题之一。基于文献综述法归纳了不同生态效益的机制及其关键影响因素，并对多效益和多影响因素进行了综合分析。结果表明，垂直绿化具有缓解城市热岛、建筑节能减排、固碳、净化空气、降噪、截留雨水和支持生物多样性等七种主要生态效益，并受到植被特征、设施结构特征、气象条件、空间格局等多方面因素的共同影响。基于对效益和影响因素的综合分析，进一步提出了两点规划思路：首先，关注环境需求和效益供给的耦合关系能够更加科学合理地指导区域垂直绿化建设布局。其次，关注生态效益间的关系，以区域关键生态环境问题为导向进行效益权衡并结合考虑其影响因素的重要性程度能为规划设计提供有效建议。     

YAP regulates periodontal ligament cell differentiation into myofibroblast interacted with RhoA/ROCK pathway

During orthodontic tooth movement (OTM), periodontal ligament cells (PDLCs) receive the mechanical stimuli and transform it into myofibroblasts (Mfbs). Indeed, previous studies have demonstrated that mechanical stimuli can promote the expression of Mfb marker α-smooth muscle actin (α-SMA) in PDLCs. Transforming growth factor β1 (TGF-β1), as the target gene of yes-associated protein (YAP), has been proven to be involved in this process. Here, we sought to assess the role of YAP in Mfbs differentiation from PDLCs. The time-course expression of YAP and α-SMA was manifested in OTM model in vivo as well as under tensional stimuli in vitro. Inhibition of RhoA/Rho-associated kinase (ROCK) pathway using Y27632 significantly reduced tension-induced Mfb differentiation and YAP expression. Moreover, overexpression of YAP with lentiviral transfection in PDLCs rescued the repression effect of Mfb differentiation induced by Y27632. These data together suggest a crucial role of YAP in regulating tension-induced Mfb differentiation from PDLC interacted with RhoA/ROCK pathway.     

H.pylori可塑区tfs3a分泌系统virB2基因功能

目的对virB2基因编码蛋白进行分析,为virB2基因及其编码蛋白功能提供实验依据。方法利用多种生物学软件以及网站对VirB2蛋白的结构和功能进行分析预测,VirB2蛋白序列通过基因推导获得并由生物公司合成,然后通过免疫动物实验制备鼠抗VirB2蛋白多克隆抗体,同时设计进行VirB2蛋白细胞毒试验(MTT法)。结果virB2基因编码蛋白属于疏水性蛋白,为鞭毛样结构,有较强的细胞毒作用。结论对VirB2蛋白的结构和功能进行了分析预测,证明VirB2蛋白在H.pylori相关的致病性特别是引起胃黏膜炎症方面起到一定的作用,能够为研究H.pylori致病机制提供帮助。     

转基因大豆MON89788芯片式数字PCR定量方法的建立

针对抗虫耐除草剂大豆转基因品系MON89788,从转基因植物基因组DNA的提取、核酸模板的质量和浓度控制、引物探针的筛选、PCR反应过程的建立等方面建立了一套完整的转基因大豆芯片式dPCR定量检测方法。本实验也对该方法的重复性和定量检测限进行考察。10组5%转基因品系大豆MON89788样品定量重复性RSD在1.17%-9.97%之间,均满足国际上转基因定量结果RSD小于25%的要求。用该方法对转基因含量为5%、1%、0.1%的大豆MON89788进行定量检测,其定量结果为5.20%、0.94%和0.11%,RSD分别为6.2%、3.6%和15.2%。该检测方法的定量限达到0.1%,能满足欧盟对转基因定量标识0.9%的要求。将本实验建立的方法用于转基因大豆的定量检测,能为规范我国转基因监管工作的实施提供强有力的技术支撑。     

Inhibition of N-linked glycosylation affects organic cation transport across the brush border membrane of opossum kidney (OK) cells.

Many organic cations are transported across the apical membrane of the proximal tubule by specific saturable mechanisms. The goal of this study was to determine if the transporter for tetraethylammonium (TEA) in the brush border membrane of an established opossum kidney (OK) cell line is glycosylated and to elucidate the function of this glycosylation. The uptake of TEA was determined in OK cell monolayers treated with tunicamycin (TM), a compound that prevents synthesis of the core oligosaccharide precursor molecules. TM exposure significantly decreased the incorporation of [3H]mannose in OK cell proteins and significantly reduced TEA uptake in a time and a concentration dependent manner. No effect of TM exposure on cellular protein synthesis, DNA content, cell viability, or on [3H]proline uptake was observed. The transport of TEA in control cells was characterized by a Km of 26.9 +/- 16.4 microM and a Vmax of 378 +/- 39 pmol/mg of protein/min. TM treatment (1 microgram/ml for 21 h) significantly increased the Km by over 4-fold to 111.5 +/- 18.4 microM while not affecting the Vmax. The apparent KI values of other organic cations known to interact with this transport system were also significantly increased by TM exposure. Estimated KI values of N1-methylnicotinamide, cimetidine, and mepiperphenidol increased by 6-fold, 4-fold, and 2-fold, respectively, after exposure of OK cells to TM. An increased KI for protons was also observed. Additional inhibitors of the N-linked glycosylation pathway, castanospermine, deoxynojirimycin, and deoxymannojirimycin significantly decreased TEA transport, whereas swainsonine had no effect. Our results suggest that the organic cation transporter is glycosylated. The N-linked oligosaccharide side chain appears to be of the hybrid type, and it either directly or indirectly affects the binding site of the transporter for both organic cations and protons. This is the first report describing the importance of glycosylation in the function of the organic cation transporter in the apical membrane of OK cells.     

尼泊尔生态系统质量评估与时空格局变化——基于参照条件的评估方法

良好的生态系统质量是维持人类社会供给需求和可持续发展目标实现的重要保障。针对尼泊尔自然地理环境复杂多样，区域间气候差异明显的特点，结合基于参照条件的评估方法可以得到生态系统质量的相对水平值，其结果能够反映出不同的变化信息。植被是区域生态系统质量变化的重要指示器，利用尼泊尔五大地理区以及四种主要植被生态系统类型划分出20个生态评估区，从表征植被生态系统的水平结构、生产功能和垂直结构3个方面计算生态参数相对密度指标（RVI），结合主成分分析法构建植被生态系统质量指数（VEQI），并以其国家自然保护区为参照，构建基于参照条件的生态系统质量评估模型，计算了尼泊尔2016和2020年基于参照条件的植被生态系统质量指数（VEQI''）并分析其生态系统质量的时空格局变化。结果表明：（1）2016至2020年，尼泊尔生态系统质量现实值VEQI的平均值增加了3.49%，总体上，在参照生态系统质量（VEQI_ref）提高（约1.41%）的背景下，生态系统质量相对水平值VEQI''增加了1.42%；（2）对于尼泊尔地区，评估区89%分位数的VEQI与其对应的国家自然保护区的参照值具有很强的相关性，总体差异较小，可以代替作为参照值；（3）从空间格局变化趋势来看，尼泊尔生态系统质量变好、基本稳定和变差的面积分别占植被生态系统总面积的74.16%、14.25%和11.59%。与数量不足、较难收集利用的野外观测台站数据相比，国家自然保护区更接近理想参照生态系统的假设，通过有限的自然保护区确定生态评估区的参照值，实现生态系统质量的快速评估，其结果具有更好的时空可比性，可以为区域生态质量变化评估及量化分析等方面提供参考。