轮状病毒感染患者外周血单个核细胞的转录组学分析

轮状病毒(Rotavirus,RV)是引起急性肠胃炎的主要病原体,分析RV感染患者的人外周血单个核细胞(Peripheral blood mononuclear cell,PBMC)中差异表达基因(Differentially expressed genes,DEGs)有利于探讨人PBMC在清除RV中的作用。为此,本研究采集2019年2月-2019年6月长春儿童医院中RV感染患者和健康儿童血液,分离PBMC,通过转录组测序(RNA sequencing,RNA-seq)技术比较RV感染患者与健康儿童之间的RNA表达图谱,借助基因本体论(Gene Ontology,GO)数据库功能富集分析、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)、Reactome通路富集分析DEGs,使用实时荧光定量PCR(Real-time quantitative PCR,qPCR)技术进行验证。结果显示,与健康对照组相比,RV感染轻症患者PBMC中有1619个DEGs;重症患者PBMC中有2816个DEGs,主要与干扰素(Interferon,IFN)反应、中性粒细胞、溶酶体、核小体、染色质等相关。qPCR验证轻症患者干扰素刺激基因(IFN-stimulated genes,ISGs)15表达上调,白介素(Interleukin,IL)1β表达下调;重症患者IL15、ISG15表达上调,IL1β表达下调,与转录组结果相一致。本研究提示,RV感染可能激活人I型和II型IFNs反应抵御病毒感染,但也会抑制溶酶体相关基因,对细胞自噬过程产生影响。     

DNA条形码在黄精属药用植物鉴定与遗传多样性分析中的应用

黄精属(Polygonatum)的许多物种具有重要的药用价值,但目前缺乏能够准确、高效地鉴定黄精属药用植物的DNA条形码。本研究通过对ITS、trnK-matK、rbcL、psbA-trnH和psbK-psbI序列进行扩增和测序,并从ITS序列中提取ITS2序列,共获得黄精属7个药用物种23份样品的138条序列。进一步比较6个DNA条形码对黄精属药用植物的鉴定效率,并验证所筛选条形码的可靠性。结果显示：trnK-matK的种内和种间变异重合少且有较明显的条形码间隙,其他5个序列的种内和种间变异重合多且无条形码间隙;BLAST结果表明trnK-matK的鉴定效率最高(85.7%),系统发育树显示trnK-matK的鉴定能力最强,能将全部12个多花黄精样品聚在一支,并能区分黄精、滇黄精、玉竹、点花黄精和湖北黄精;AMOVA分析结果揭示trnK-matK的群体遗传分化指数(F_st)最高,适用于区分黄精属物种间差异。因此,trnK-matK最适用于黄精属药用植物的分子鉴定。     

森林植物多样性、树种重要值与土壤理化性质对球囊霉素相关土壤蛋白的影响

球囊霉素相关土壤蛋白(glomalin-related soil protein, GRSP)在土壤物理结构调节和土壤碳库稳定性中发挥着重要作用,但植物多样性和优势种如何影响GRSP还缺乏系统性研究。本研究依托东北林业大学哈尔滨实验林场的72块样地, 对1 m深土壤剖面分5层采样, 测定土壤易提取球囊霉素(easily extractable GRSP, EEG)、总提取球囊霉素(total GRSP, TG)及土壤理化性质, 并同时计算植物多样性指数及优势种重要值(importance value, IV), 进一步通过相关分析和冗余排序分析判断影响GRSP的主要因素与贡献。结果表明: (1)在整个土壤剖面上均表现为TG和EEG与土壤有机碳(SOC)正相关, 在部分土层深度与全氮(total nitrogen, TN)和含水量(moisture content, MC)正相关, 而与电导率(electrical conductivity, EC)和pH值负相关。(2)部分土层TG和EEG与黑皮油松(Pinus tabuliformis var. mukdensis)、樟子松(P. sylvestris var. mongolica)、胡桃楸(Juglans mandshurica)、黄檗(Phellodendron amurense)、榆树(Ulmus pumila)优势种重要值显著相关, 表现为黑皮油松重要值越高, 而黄檗、榆树重要值越小, 越有利于EEG的积累, 并且伴随EEG-C/SOC (EEG中C占SOC比例)增加、EEG/TG增大; 群落中胡桃楸、黄檗、榆树更有利于TG积累, 黑皮油松、落叶松(Larix gmelinii)、樟子松不利于TG的积累。(3)植物Simpson指数、Shannon-Wiener指数、物种丰富度与EEG、TG、EEG/TG无显著相关性, 而与EEG-C/SOC、EEG-N/TN (EEG中N占TN的比例)、TG-C/SOC (TG中C占SOC比例)、TG-N/TN (TG中N占TN的比例)显著负相关; 土壤EEG/TG和EEG-N/TN与植物均匀度指数显著正相关, 在1 m土壤不同土层趋势类似。(4)方差分解分析表明: 生物因子对GRSP变化的解释率是20.2%, 土壤理化因子解释率为7.8%, 而生物因子中植物优势种重要值的解释率最大(16.4%), 而植物物种多样性指数解释率仅为0.4%。冗余排序发现常绿针叶树种(黑皮油松和樟子松)越多且阔叶树种越少时, GRSP含量和GRSP对土壤碳氮的贡献越高(P < 0.01), 其机制可能与树种菌根类型有关: 外生菌根树种重要值与TG显著负相关, 丛枝菌根树种重要值与TG显著正相关。本研究解析了植物物种多样性对GRSP含量的重要影响, 并强调未来土壤管理和评估可以通过调整优势物种而不是树种多样性来促进GRSP积累。     

小兴安岭凉水国家级自然保护区植物beta多样性及其影响因素

自然保护区如何设置才能够最大程度保护生物多样性, 是保护生物学的研究热点; 阐明beta多样性特征、组分格局及其影响因素是保护生物学的重要基础。本研究选取小兴安岭凉水国家级自然保护区不同功能区(核心区、缓冲区、实验区)及毗邻地区(保护区外)共80块样方作为研究对象, 调查每块样方的保护位置(经纬度、海拔、坡位、坡度、坡向)和群落结构(郁闭度、林龄、乔木树高、胸径、灌木树高、地径), 并采集0-20 cm土壤样品, 测定土壤理化性质(有机碳、全氮、pH值、电导率、含水量、容重)。将样方间的beta多样性分解为物种周转和物种多度差异两种组分, 通过Mantel分析、冗余分析和方差分解分析解析非生物因子(地理地形、保护强度、土壤因子)和生物因子(群落结构)对beta多样性及其组分的影响。结果表明: (1)乔、灌、草3层中, 物种周转组分对于beta多样性的贡献均占主导地位(65%-73%), 物种多度差异贡献较小。(2) Mantel检验结果表明, 乔、灌、草3层beta多样性及其组分与地理地形指标显著相关的因子最多; 土壤因子只对乔木层和灌木层beta多样性及组分有影响, 对草本层影响不大。其中坡位、坡度、乔木树高和保护强度均与保护区乔、灌、草3层beta多样性显著正相关(P < 0.05)。(3)植物整体beta多样性受地理地形影响最大, 但存在乔、灌、草差异。乔木层beta多样性受生物因子影响最大; 灌木层的土壤因子解释力分别为地理地形和生物因子的2倍; 而草本层主要受地理地形的影响, 其解释力分别是土壤和生物因子的26倍和3倍。乔木胸径对植物beta多样性差异具有最大的解释作用。本研究结果表明, 未来保护区设置需要根据保护植物的类型, 选择适当的林分结构、土壤和地理地形等, 以增强保护区植物多样性保护的效果。     

锶污染下燕麦对土壤酶活性和微生物群落功能多样性的影响

为了评价燕麦对金属锶(Sr)污染土壤的修复效果,通过土培盆栽试验,在土壤中添加Sr,共设置6个处理:25 mg/kg Sr(CK)、100 mg/kg Sr(L)、250 mg/kg Sr(H)、25 mg/kg Sr+燕麦(CK+燕麦)、100 mg/kg Sr+燕麦(L+燕麦)和250 mg/kg Sr+燕麦(H+燕麦)。从土壤酶(转化酶、磷酸酶、蛋白酶、脲酶和脱氢酶)活性和土壤微生物群落功能多样性来评价燕麦在锶污染土壤植物修复中的作用。结果表明:燕麦显著提高了脲酶和脱氢酶活性。燕麦显著提高了锶污染土壤微生物对31种碳源的平均利用率(AWCD),并提高了锶污染土壤条件下微生物群落的Shannon多样性指数(H)、Shannon均匀度指数(E)和碳源利用丰富度指数(S),降低了Shannon优势度指数(D)。主成分分析表明种植燕麦显著增加了锶污染土壤中微生物群落功能多样性。与未种植处理(L和H)相比,L+燕麦处理中羧酸类碳源和聚合物类碳源的利用率分别增加了19%和31%,H+燕麦处理中羧酸类碳源和聚合物类碳源的利用率分别增加了24%和41%。因此,在锶污染土壤中,燕麦显著提高了土壤酶活性,改善了微生物功能多样性。     

Altering Neurospora crassa MOB2A exposes its functions in development and affects its interaction with the NDR kinase COT1

The Neurospora crassa Mps One Binder (MOB) proteins MOB2A and MOB2B physically interact with the Nuclear Dbf2 Related (NDR) kinase COT1 and have been shown to have overlapping functions in various aspects of asexual development. Here, we identified two N. crassa MOB2A residues, Tyr117 and Tyr119, which are potentially phosphorylated. Using phosphomimetic mob‐2a mutants we have been able to establish that apart from their previously described roles, MOB2A/B are involved in additional developmental processes. Enhanced conidial germination, accompanied by conidial agglutination, in the phosphomimetic mutants indicated that MOB2A is a negative regulator of germination. Thick‐section imaging of perithecia revealed slow maturation and a lack of asci alignment in the mutant strains demonstrating a role for MOB2A in sexual development. We demonstrate that even though MOB2A and MOB2B have some overlapping functions, MOB2B cannot compensate for the roles MOB2A has in conidiation and germination. Altering Tyr residues 117 and 119 impaired the physical interactions between MOB2A and COT1, most likely contributing to some of the observed effects. As cot‐1 and the phosphomimetic mutants share an extragenic suppressor (gul‐1), we concluded that at least some of the effects imposed by altering Tyr117 and Tyr119 are mediated by the NDR kinase.     

Influence of Polymers on the Physical and Chemical Stability of Spray-dried Amorphous Solid Dispersion: Dipyridamole Degradation Induced by Enteric Polymers

Amorphous solid dispersions (ASDs) are inherently unstable because of high internal energy. Evaluating physical and chemical stability during the process and storage is essential. Numerous researches have demonstrated how polymers influence the drug precipitation and physical stability of ASDs, while the influence of polymers on the chemical stability of ASDs is often overlooked. Therefore, this study aimed to investigate the effect of polymers on the physical and chemical stability of spray-dried ASDs using dipyridamole (DP) as a model drug. Proper polymers were selected by assessing their abilities to inhibit drug recrystallization in supersaturated solutions. HPMC E5, Soluplus®, HPMCP-55, and HPMCAS-LP were shown to be effective stabilizers. The optimized formulations were further stored at a high temperature (60 °C) and high humidity (40 °C, 75% RH) for 2 months, and their physical and chemical stability was evaluated using polarizing optical microscopy, FTIR, HPLC, and mass spectrometry (MS). In general, crystallization was observed in all samples, which indicated the physical instability under stressed storage conditions. Also, it was noted that the polymers in ASDs rather than physical mixtures, induced a dramatic drug degradation after being exposed to a high temperature (HPMCP-55 >?80% and HPMCAS-LP >?50%) and high humidity (HPMCP-55 >?40% and HPMCAS-LP >?10%). The MS analysis further confirmed the degradation products, which might be generated from the reaction between dipyridamole and phthalic anhydride decomposed from HPMCP-55 and HPMCAS-LP. Overall, the exposure of ASDs to stressed conditions resulted in recrystallization and even the chemical degradation induced by polymers.