Large-scale identification of proteins in human salivary proteome by liquid chromatography/mass spectrometry and two-dimensional gel electrophoresis-mass spectrometry

Human saliva contains a large number of proteins and peptides (salivary proteome) that help maintain homeostasis in the oral cavity. Global analysis of human salivary proteome is important for understanding oral health and disease pathogenesis. In this study, large-scale identification of salivary proteins was demonstrated by using shotgun proteomics and two-dimensinal gel electrophoresis-mass spectrometry (2-DE-MS). For the shotgun approach, whole saliva proteins were prefractionated according to molecular weight. The smallest fraction, presumably containing salivary peptides, was directly separated by capillary liquid chromatography (LC). However, the large protein fractions were digested into peptides for subsequent LC separation. Separated peptides were analyzed by on-line electrospray tandem mass spectrometry (MS/MS) using a quadrupole-time of flight mass spectrometer, and the obtained spectra were automatically processed to search human protein sequence database for protein identification. Additionally, 2-DE was used to map out the proteins in whole saliva. Protein spots 105 in number were excised and in-gel digested; and the resulting peptide fragments were measured by matrix-assisted laser desorption/ionization-mass spectrometry and sequenced by LC-MS/MS for protein identification. In total, we cataloged 309 proteins from human whole saliva by using these two proteomic approaches.     

Bioleaching of chalcopyrite concentrate by a moderately thermophilic culture in a stirred tank reactor

A mixed culture of moderately thermophilic microorganisms was enriched from acid mine drainage samples collected from several chalcopyrite mines in China. Such mixed culture can be used to effectively extract copper from chalcopyrite. Furthermore, after being adapted to gradually increased concentration of chalcopyrite concentrate, the tolerance of the mixed culture to chalcopyrite concentrate was brought up to 80 g/L. The effects of several leaching parameters on copper recovery in stirred tank reactor also had been investigated. The results of the investigation show that it was possible to achieve a copper extraction rate of 75% in 44 days at a pulp density of 8%. The leaching rate of chalcopyrite concentrate tended to increase with dissolved total iron concentration. At low pH ranges, more microscopic counts of microorganisms were found in the solution. Furthermore, the analysis of leached residues indicates that the passivation of chalcopyrite concentrate was mainly due to a mass of jarosite and PbSO(4) on the mineral surface, other than the elemental sulphur layer. The bacterial community composition was analyzed by using Amplified Ribosomal DNA Restriction Analysis. Two moderately thermophilic bacteria species were identified as Leptospirillum ferriphilum and Acidithiobacillus caldus with abundance of 67% and 33% in the bio-pulp, respectively.     

Interleukin-33/ST2 signaling promotes production of interleukin-6 and interleukin-8 in systemic inflammation in cigarette smoke-induced chronic obstructive pulmonary disease mice

Interleukin-33 is a newly described member of the interleukin-1 family. Recent research suggests that IL-33 is increased in lungs and plays a critical role in chronic airway inflammation in cigarette smoke-induced chronic obstructive pulmonary disease (COPD) mice. To determine the role of IL-33 in systemic inflammation, we induced COPD mice models by passive cigarette smoking and identified the IL-33 expression in bronchial endothelial cells and peripheral blood mononuclear cells (PBMCs) of them. After isolation, PBMCs were cultured and stimulated in vitro. We measured expressions of interleukin-6 and interleukin-8 in PBMCs in different groups. The expression of IL-33 in bronchial endothelial cells and PBMCs of COPD mice were highly expressed. Stimulated by cigarette smoke extract (CSE), the expression of IL-6 and IL-8 were induced and enhanced by IL-33. PBMCs of COPD mice produced more IL-6 and IL-8 stimulated by CSE and IL-33. Expression of IL-6 and IL-8 were decreased when stimulated by IL-33 together with soluble ST2. The mRNA production of ST2 in IL-33 stimulated PBMCs was increased. Being pretreated with several kinds of MAPK inhibitors, the secretions of IL-6 and IL-8 in PBMCs did not decrease except for the p38 MAPK inhibitor. We found that IL-33 could induce and enhance the expression of IL-6 and IL-8 in PBMCs of COPD mice via p38 MAPK pathway, and it is a promoter of the IL-6 and IL-8 production in systemic inflammation in COPD mice.     

LC-MS/MS determination of helicid in human plasma and its application in pharmacokinetic studies

Helicid is a traditional Chinese medicine used to treat headache and insomnia with definite effects. To facilitate pharmacokinetic studies of helicid in man, a sensitive and specific LC-MS/MS method for the quantitative detection of helicid in human plasma was developed and validated. The method involved the addition of bergeninum as the internal standard (IS), protein precipitation, HPLC separation, and quantification by MS/MS system using negative electrospray ionization in the multiple reaction monitoring mode (MRM). The precursor→product ion transitions were monitored at m/z 282.8→120.9 for helicid and m/z 326.9→192.2 for the IS, respectively. The lower limit of quantification (LLOQ) was 0.2 μg/L. The calibration curves for helicid was linear over a concentration range of 0.2-20 μg/L. The intra- and inter-batch analyses of QC samples at 0.4, 2, 20 μg/L indicated good precision (%R.S.D. between 2.69 and 5.47%) and accuracy (between 96.15 and 105.05%). The helicid was stable in human plasma stored at room temperature for at least 24h, 4°C for at least 24h, -20°C for at least 1 month, and for routine three freeze-thaw cycles. This accurate and specific assay provides a useful method for evaluating the pharmacokinetic profile of helicid in humans.     

Five New Guaiane Sesquiterpenes from the Endophytic Fungus Xylaria sp. YM 311647 of Azadirachta indica

Five new guaiane sesquiterpenes, 1 – 5 , were isolated from the culture broth of the endophytic fungus Xylaria sp. YM 311647, isolated from Azadirachta indica A. Juss . The structures of these compounds were elucidated on the basis of spectroscopic analyses, and their inhibitory activities against five pathogenic fungi were evaluated. All guaiane sesquiterpenes showed moderate or weak antifungal activities in a broth microdilution assay.     

江西省野生维管植物名录

为了全面了解江西省野生维管植物资源的现状并为开展相应的生物多样性保护实践提供基础资料, 本文在前人的基础上, 通过进一步的文献收集、标本考证、数据分析, 并结合作者的野外调查结果, 按照维管植物最新的分类系统整理出江西省野生维管植物名录, 并根据标本信息核实了每个物种县市级的分布信息。结果显示, 江西省共有野生维管植物214科1,253属4,761种(含种下等级), 其中石松类与蕨类植物35科103属444种, 裸子植物5科21属36种, 被子植物174科1,129属4,281种。石松类与蕨类植物中, 物种数最多的前5个科为鳞毛蕨科(84种)、水龙骨科(55种)、凤尾蕨科(49种)、蹄盖蕨科(47种)和金星蕨科(47种), 最多的属是鳞毛蕨属(Dryopteris, 40种); 裸子植物中物种数最多的科和属为松科(7属12种)和松属(Pinus, 5种); 被子植物是江西省野生维管植物的最主要组成部分, 分别占科、属、种总数的81.3%、90.1%和89.9%, 其中种数最多的前5个科分别是禾本科(315种)、菊科(241种)、蔷薇科(228种)、唇形科(200种)和豆科(197种), 前6个属依次是薹草属(Carex, 71种)、悬钩子属(Rubus, 66种)、冬青属(Ilex, 56种)、蓼属(Persicaria, 40种)、珍珠菜属(Lysimachia, 37种)和杜鹃花属(Rhododendron, 37种)。江西省维管植物调查呈现出地区不均衡性, 建议增加后续调查工作的广度和深度, 尤其在武夷山脉、赣南山区、鄱阳湖湿地等生物多样性较高或较特殊的地区进行补充采集。同时, 本文呼吁加强专科专属的分类学研究, 为后续名录的修订与完善奠定基础。     

长白山垂直自然带弹尾纲物种概况

以长白山弹尾纲为研究对象, 基于1758‒2022年度发表的分类学和生态学文献资料, 编制了长白山垂直自然带弹尾纲物种名录, 分析了首次发表物种的学者贡献、出版物、发表时间和各属沿海拔梯度的分布情况。本研究共梳理弹尾纲物种92种, 隶属于3目11科46属, 长角䖴目、原䖴目和愈腹䖴目的物种分别占比61%、31%和8%, 其中长角䖴目的等节䖴科(该类群物种数占总体比例, 26%)、长角䖴科(16%)和鳞䖴科(16%)、原䖴目的棘䖴科(26%)是该地区的主要类群。统计的92种长白山弹尾纲物种由36位学者首次定名和报道, 中国学者占主要贡献(33%), 其次为波兰(11%)、瑞典(11%)、俄罗斯(8%)和美国(8%); 相关论文多发表在国际期刊上, Zootaxa是发表的主要期刊。基于海拔梯度的统计发现, 鳞䖴属(Tomocerus)和符䖴属(Folsomia)的物种沿海拔梯度的分布范围最广(海拔800‒1,700 m), 分布在海拔800 m (15个物种)、1,100 m (20个物种)和1,400 m (14个物种)的物种最多。基于以上结果, 本文讨论了长白山弹尾纲分类学现状和发展前景。     

基于光合系统参数建立马铃薯耐荫性综合评价体系

为构建便捷的马铃薯(Solanum tuberosum)耐荫性综合评价体系并发掘耐荫种质, 以35个马铃薯品种(系)为实验材料, 测定块茎膨大期遮荫下植株叶片叶绿素含量、光合能力和叶绿素荧光等光合参数及收获后块茎单株产量和淀粉含量等指标。根据耐荫系数, 利用主成分分析法、隶属函数法、聚类分析法和逐步回归分析法进行综合评价。通过主成分分析将马铃薯耐荫性相关的13个单项光合指标转换为6个综合指标, 代表了全部信息的87.51%。以此计算各种质的隶属函数值, 并以主成分的贡献率进行加权, 最终获得所用材料耐荫性的综合评价值(D值)。根据D值聚类分析结果将35个马铃薯分为4类, 其中Eshu10和Lishu6分别为耐荫性最强和最弱的品种。通过逐步回归分析建立了马铃薯耐荫性评价数学模型: D=0.060+0.106G_s+0.214qP+0.143NPQ。同时, 用该评价体系鉴定为耐荫性强的品种(系)在遮荫后其产量和/或淀粉含量等指标减幅均低于耐荫性弱的种质, 表明该评价体系可用于快速评价和预测马铃薯种质的耐荫性。     

苏云金芽孢杆菌S1478-1分离株的特性鉴定及cry1Ac同源基因克隆

本研究对从海南岛尖峰岭热带雨林自然保护区的土壤样品中分离出的Bt菌株S1478-1进行了特性鉴定,研究表明S1478-1分离株菌落形态和生长特征和Bt参照菌株HD73极其相似.16S rDNA序列分析表明,S1478-1分离株与其它B.thuringiensis、B.cereus和B.anthracis的16S rDNA序列相似性达到99%.分离株能产菱形伴胞晶体,SDS-PAGE蛋白电泳分析表明,菌株在生长后期,形成芽孢同时分泌130 kD大小的晶体蛋白.生物测定表明S1478-1分离株对小菜蛾具有很高的毒杀活性,LC50卯值高达5.159 ×108cfu/mL.初步显示S1478-1分离株可作为防治鳞翅目害虫的生物农药菌株.利用PCR-RFLP方法鉴定S1478-1分离株含有cry1Ac同源基因,以PCR粘性端克隆方法扩增全长基因,序列测定表明该基因ORF为3 537bp,编码1178个氨基酸,推定的编码蛋白分子量为133.3 kD,与其它cry1Ac基因序列最高达到99%同源,因此,该基因可作为杀虫工程菌及培育转基因抗虫作物的候选基因.     

秤锤树属与长果安息香属植物的地理分布及其濒危现状

秤锤树属(SinojackiaHu)和长果安息香属(ChangiostyraxC.T.Chen)是安息香科的少种属,这两属在我国共记录有7个种。本文通过野外调查,分析了中国这两属植物的地理分布、濒危现状及其迁地保护状况。结果表明:秤锤树属植物地理分布较广,但是每个物种的居群数量和居群大小均很小。其中秤锤树(Sinojackiaxylocarpa)和狭果秤锤树(S.rehderiana)已经在其模式标本产地灭绝;棱果秤锤树(S.henryi)在过去的近70年内没有采到过标本,该物种可能存在同物异名现象或已经灭绝;细果秤锤树(S.microcarpa)由于人为破坏严重,居群大小急剧下降;肉果秤锤树(S.sarcocarpa)和怀化秤锤树(S.oblongicarpa)呈零星分布且个体数量很少,处于极濒危状态。另外本次调查发现秤锤树属的一个新的分类群(待鉴定种)。秤锤树属的大多数种和长果安息香属植物的居群更新能力差:虽然结果率较高,但是结籽率较低;坚硬的内果皮阻碍了种子的萌发,这是其居群更新的最大障碍;另外人为破坏对其居群更新的影响也较大。作者建议应该把秤锤树属的所有物种和长果安息香属植物都纳入保护的范围并讨论了这两属植物的保护策略。