Schiff bases of tryptamine as potent inhibitors of nucleoside triphosphate diphosphohydrolases (NTPDases): Structure-activity relationship

Overexpression of NTPDases leads to a number of pathological situations such as thrombosis, and cancer. Thus, effective inhibitors are required to combat these pathological situations. Different classes of NTPDase inhibitors are reported so far including nucleotides and their derivatives, sulfonated dyes such as reactive blue 2, suramin and its derivatives, and polyoxomatalates (POMs). Suramin is a well-known and potent NTPDase inhibitor, nonetheless, a range of side effects are also associated with it. Reactive blue 2 also had non-specific side effects that become apparent at high concentrations. In addition, most of the NTPDase inhibitors are high molecular weight compounds, always required tedious chemical steps to synthesize. Hence, there is still need to explore novel, low molecular weight, easy to synthesize, and potent NTPDase inhibitors.Keeping in mind the known NTPDase inhibitors with imine functionality and nitrogen heterocycles, Schiff bases of tryptamine, 1–26, were synthesized and characterized by spectroscopic techniques such as EI-MS, HREI-MS, ¹H-, and ¹³C NMR. All the synthetic compounds were evaluated for the inhibitory avidity against activities of three major isoforms of NTPDases: NTPDase-1, NTPDase-3, and NTPDase-8. Cumulatively, eighteen compounds were found to show potent inhibition (Ki = 0.0200–0.350 μM) of NTPDase-1, twelve (Ki = 0.071–1.060 μM) of NTPDase-3, and fifteen compounds inhibited (Ki = 0.0700–4.03 μM) NTPDase-8 activity. As a comparison, the Kis of the standard inhibitor suramin were 1.260 ± 0.007, 6.39 ± 0.89 and 1.180 ± 0.002 μM, respectively. Kinetic studies were performed on lead compounds (6, 5, and 21) with human (h-) NTPDase-1, -3, and -8, and Lineweaver-Burk plot analysis showed that they were all competitive inhibitors. In silico study was conducted on compound 6 that showed the highest level of inhibition of NTPDase-1 to understand the binding mode in the active site of the enzyme.     

安徽贝母属植物叶片的比较解剖学研究

在光学显微镜下,对安徽省贝母属(Fritillaria L.)8种1变种,其中包括安徽贝母(F. anhuiensis)和宁国贝母(F. ningguoensis)各两个居群的植物叶片进行了解剖结构观察,结果发现：叶肉细胞均含有淀粉粒,近上表皮的叶肉细胞多为方形或圆柱状,较大,排列较紧密,近下表皮的叶肉细胞形状不规则,多有分枝,细胞间隙大;叶均无表皮毛等附属物,除铜陵黄花贝母(F. monantha var. tonglingensis)的下表皮细胞类似平行四边形外,其它种叶表皮细胞表面观为长方形或长条形,细胞长轴与叶脉平行,垂周壁为平直、浅波状或深波状;气孔器为Allium型,仅分布于下表皮,椭圆形,长轴与叶脉平行,单个随机分布,部分种保卫细胞两极有“T”形加厚。叶表皮细胞的形状,垂周壁式样,气孔器的类型等特征较为稳定,可以为贝母属内种的划分及系统演化关系的探讨提供实验证据。     

Genetic diversity of the endangered species Kirengeshoma palmata (Saxifragaceae) in China

In order to investigate the levels of genetic diversity of the endangered species Kirengeshoma palmata (Saxifragaceae), four extant populations were sampled and analyzed using inter-simple sequence repeats (ISSR) markers. We expected a low genetic diversity level, but our results revealed a high level of intraspecific genetic diversity, probably resulting from this species being in a refuge during the last glaciation (at population level: P = 63.25%, A_e = 1.47, H_E = 0.26 and H_O = 0.37; at species level: P = 79.00%, A = 1.5538, H_T = 0.2586 and H_sp = 0.3104). A low level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (16.69%) and AMOVA (19.36%). Populations shared high levels of genetic identity. Insect pollination and seed dispersal by wind may have facilitated extensive gene flow and are likely responsible for this present structure of genetic variation.     

黄山濒危植物小花木兰生态位与年龄结构研究

基于对黄山风景区濒危植物小花木兰（Magnolia seiboldii）分布群落的样地调查,采用定量分析的方法,研究了小花木兰群落中主要种群的生态位宽度、生态位重叠和小花木兰种群的年龄结构．结果表明：（1）分别用Levins和Shannon weiner两指数测得的主要优势种群的生态位宽度结果基本一致;重要值大的种群一般具有较宽的生态位。生态位重叠与生态位宽度有关,具较宽生态位的种群一般与其他种群具有较高的生态位重叠,另外,具有相似生境要求和相似生物学特征的种群间也具有较大的生态位重叠,如都是耐萌种的小花木兰与四照花（Dendrobenthamia japonica）的生态位重叠值较高。种群间生态位重叠程度较高,物种间关系复杂,对资源共享趋势明显。（2）小花木兰为星散间断分布,在群落中处于伴生从属地位,生存易受群落变化的影响。对生境要求相似的种群在同一生境下当资源充分时能形成良好的共生关系,支持林思祖等人的生态位重叠与竞争不成相关关系的学说。（3）通过对小花木兰年龄结构的统计发现,小花木兰年龄结构不完整,属于衰退型,幼苗储备严重不足,成为该种群更新的一大瓶颈。（4）它的高海拔分布显示了低温是它生存的一个重要保证;水分也是影响小花木兰种群动态的重要因子。散花坞生境中小花木兰生长良好,呈增长趋势,而清凉台生境小花木兰则表现为年龄老化,呈衰退趋势。散花坞和清凉台可视为重点保护生境,其间生境差异导致的种群动态差异有待于进一步探讨。     

基于HNSOTER的海南岛土壤有机碳储量及空间分布特征分析

基于海南岛1∶200 000土壤 地体数字化数据库（HNSOTER）,在GIS系统的支持下,对海南岛土壤有机碳储量及分布特征进行了探讨.结果表明,1）标准剖深下（0～100 cm）,海南岛土壤有机碳储量为2.78×10⁸ t.2）0～20 cm剖深土壤有机碳密度变幅在0.3～18.8 kg·m^-2之间,其中1.0～5.0 kg·m^-2密度区占总分布面积的81.2%,按面积加权均值为3.3 kg·m^-2.0～100 cm剖深的土壤有机碳密度变幅在1.0～32.1 kg·m^-2 之间,其中2.0～14.0 kg·m^-2密度区面积比重占89.7%,按面积加权均值为8.4 kg·m^-2.不同地形、岩性及土壤类型中,土壤有机碳密度分布有较大变异.3）从中部山地向外至沿海平原,土壤有机碳密度总体上呈递减趋势,但不同剖深下其分布格局仍有一定差异.0～20 cm剖深下土壤有机碳密度高值区（丰富度指数大于1）主要分布于山地以及北部玄武岩台地,0～100 cm剖深下,有机碳密度高值区（丰富度指数大于1）趋向集中于中东部山地、台地区,且其分布重心较前者明显的向东偏移.     

咸鸭蛋清胃蛋白酶水解产物的抗菌性及抗氧化性

【目的】提高对咸鸭蛋清的综合利用,制备具有生物活性的蛋白水解产物。【方法】采用超滤对咸鸭蛋清进行脱盐及分级,利用胃蛋白酶对不同分子量范围的咸鸭蛋清蛋白组分进行酶解,并对不同组分的水解产物进行体外抑菌试验,测定其最低抑菌浓度(MIC)、最低杀菌浓度(MBC)、细菌细胞膜完整性及抗氧化活性。【结果】超滤后得到3种不同分子量的组分,组分A (Mw>50 kDa)、组分B (50 kDa>Mw>20 kDa)和组分C (20 kDa>Mw>5 kDa)。其中,组分A和组分B的水解产物对大肠杆菌的生长有抑制作用。2种组分的MIC分别为1 024 μg/mL和2 048 μg/mL,MBC均为2 048 μg/mL。大肠杆菌进入稳定期OD₆₀₀值约为1.15,添加2种水解产物浓度达1×MIC时,OD₆₀₀值降至0.79和0.86,浓度达2×MIC时,OD₆₀₀值降至0.50和0.68。2种水解产物可破坏大肠杆菌细胞膜,达到较好的抑菌效果。而金黄色葡萄球菌对3种水解产物均不敏感,抑菌效果较差。另外,3种水解产物均具有抗氧化活性,它们对DPPH自由基清除能力分别达0.5倍Trolox (水溶性维生素E)、0.67倍Trolox、0.38倍Trolox。【结论】超滤可同时实现咸蛋清脱盐及富集不同目标蛋白的目的。目标蛋白酶解组分具备抗菌活性和抗氧化活性,同时可作为一种营养添加剂,提高了咸鸭蛋清的附加价值和利用率。     

The Effect of α7nAChR Signaling on T Cells and Macrophages and Their Clinical Implication in the Treatment of Rheumatic Diseases

Neurochemical Research - Rheumatoid arthritis (RA) is one of the most common autoimmune disease and until now, the etiology and pathogenesis of RA is not fully understood, although dysregulation of...     

利用CRISPR/Cas9系统构建稳定敲除TrxR1基因的HCT-116细胞株北大核心CSCD

为更好地研究靶向硫氧还蛋白还原酶1的小分子化合物的细胞内靶点选择性,利用CRISPR/Cas9系统构建稳定敲除TrxR1基因(编码硫氧还蛋白还原酶1)的HCT-116细胞株。首先根据TrxR1基因序列和CRISPR/Cas9靶点设计原则,设计并选择合适的敲除位点,再根据敲除位点序列设计敲除TrxR1基因的sgRNA干扰序列,以pCasCMV-Puro-U6空质粒载体为骨架构建能表达该sgRNA干扰序列的重组质粒。质粒共转染至HCT-116细胞后,利用嘌呤霉素筛选TrxR1敲除的HCT-116细胞,通过DNA测序、免疫蛋白印迹、TRFS-green荧光探针和细胞内TrxR1酶活力检测等方法鉴定和验证HCT-116细胞的TrxR1基因敲除效果。进一步通过CCK-8实验初步研究靶向TrxR1小分子化合物对细胞内TrxR1酶活力和细胞增殖力抑制的相关性。结果显示,表达sgRNA干扰序列的重组质粒可以敲除HCT-116细胞中TrxR1基因,筛选获得的稳定敲除细胞HCT116-TrxR1-KO中无TrxR1蛋白表达,而靶向TrxR1小分子抑制剂对该细胞无TrxR1酶活力和细胞增殖力抑制效果。本研究利用CRISPR/Cas9系统成功构建了HCT-116的TrxR1基因敲除的稳定细胞株,为进一步研究TrxR1在相关疾病的发生机制和治疗中的作用奠定了基础。     

Protein engineering of Bacillus acidopullulyticus pullulanase for enhanced thermostability using in silico data driven rational design methods

Thermostability has been considered as a requirement in the starch processing industry to maintain high catalytic activity of pullulanase under high temperatures. Four data driven rational design methods (B-FITTER, proline theory, PoPMuSiC-2.1, and sequence consensus approach) were adopted to identify the key residue potential links with thermostability, and 39 residues of Bacillus acidopullulyticus pullulanase were chosen as mutagenesis targets. Single mutagenesis followed by combined mutagenesis resulted in the best mutant E518I-S662R-Q706P, which exhibited an 11-fold half-life improvement at 60 °C and a 9.5 °C increase in T_m. The optimum temperature of the mutant increased from 60 to 65 °C. Fluorescence spectroscopy results demonstrated that the tertiary structure of the mutant enzyme was more compact than that of the wild-type (WT) enzyme. Structural change analysis revealed that the increase in thermostability was most probably caused by a combination of lower stability free-energy and higher hydrophobicity of E518I, more hydrogen bonds of S662R, and higher rigidity of Q706P compared with the WT. The findings demonstrated the effectiveness of combined data-driven rational design approaches in engineering an industrial enzyme to improve thermostability.     

两湖泊轮虫群落结构的时空动态对水体氮磷含量及“下行效应”的响应

为探讨氮和磷、"上行效应"和"下行效应"在轮虫群落结构时空变动调控中的相对重要性,于2011年7月至2012年6月每月两次采集了芜湖市九莲塘和汀棠湖中的轮虫样品,利用相关分析法和典范对应分析法(CCA)分析了轮虫群落结构的时空变动与水体理化因子及其潜在捕食者密度间的关系。卡尔森营养状态指数表明,两湖均处在富营养化初期。除了总氮和硝态氮的年均浓度、晶囊轮虫(Asplanchna)和剑水蚤(Cyclopoidea)无节幼体的年均密度存在显著差异外(P0.05),两湖泊的其他水环境因子间并未表现出明显的差异(P0.05)。实验室鉴定发现九莲塘中共有轮虫48种,隶属于15科21属;汀棠湖中共有轮虫55种,隶属于15科24属。两湖泊中,疣毛轮虫(Synchaeta)、臂尾轮虫(Brachionus)和异尾轮虫(Trichocerca)均是主要轮虫类群,而密度优势种分别有7种和4种。相关分析结果显示,P含量与九莲塘轮虫群落的均匀度和物种多样性指数间均呈现显著的负相关(P0.05),而N含量却与汀棠湖轮虫群落的物种多样性指数呈现显著的正相关(P0.05);两湖泊中的轮虫物种多样性指数均与水体中的TN∶TP(质量比)间呈现显著的正相关(P0.05)。CCA分析结果显示,九莲塘中桡足类和其无节幼体密度显著高于其他采样批次时的轮虫群落聚成Ⅰ类,汀棠湖中晶囊轮虫密度显著高于其他批次时的轮虫群落聚成Ⅱ类,而其他的轮虫潜在捕食者密度很低或未出现时的轮虫群落聚成Ⅲ类。研究结果表明,湖泊中的TN∶TP(质量比)可能是反映N和P含量对轮虫群落物种多样性影响程度的较为合适的指标。当水体中的轮虫潜在捕食者密度较高时,两湖泊轮虫群落结构的变动均主要取决于"下行效应";而当轮虫潜在捕食者密度很低时,"上行效应"可能是调控两湖泊轮虫群落结构变动的主要因素。