当归赤芍药对中阿魏酸和没食子酸在大鼠血浆和尿液中的代谢物研究

目的:研究当归赤芍药对中阿魏酸和没食子酸在大鼠血浆和尿液中的代谢物。方法:大鼠灌胃当归赤芍药对后,采用UPLC-Q-TOF-MS联用技术,对其血浆及尿液中阿魏酸和没食子酸的代谢物进行分析。结果:根据质谱信息,推测阿魏酸在大鼠体内的代谢物主要以甲基化、硫酸结合、葡萄糖醛酸结合等形式存在;没食子酸则主要以还原、甲基化硫酸结合、甲基化葡萄糖醛酸结合等形式存在。结论:当归赤芍药对中阿魏酸及没食子酸在大鼠体内存在多种Ⅰ,Ⅱ相代谢产物,为阐明该药对在体内发挥疗效的药效物质基础和作用机制提供依据。     

产黄青霉Penicillium chrysogenum S-3-25人工海水培养基发酵次级代谢产物研究

目的 阐明南极深海来源真菌产黄青霉Penicillium chrysogenum S-3-25人工海水培养基的发酵次级代谢产物及其活性。方法 利用各种色谱技术分离纯化次级代谢产物,根据理化和波谱数据（核磁共振、质谱技术和Marfey分析）鉴定化合物结构,采用3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide（MTT）法评价细胞毒活性。结果 从真菌S-3-25人工海水培养基发酵产物中分离鉴定了8个单体化合物：2-[[(2S)-2-amino-1-oxopropyl] amino] benzoic acid (1),methyl 2-[[(2S)-2-amino-1-oxopropyl] amino] benzoate (2),2-[[(2S)-2-hydroxy-1-oxopropyl] amino] benzamide (3),3-(p-hydroxyphenyl)-N-methylpropionamide (4),4-hydroxycinnamamide (5),cyclo-(L-Pro-L-Tyr) (6),脑苷脂A (7) 及脑苷脂B (8)。细胞毒活性测试结果表明化合物1~8在50 μM作用浓度下对三种受试细胞（人乳腺癌MCF-7,人肺癌A549以及小鼠小胶质BV2细胞）的抑制率均低于50%。结论 从极地深海来源真菌产黄青霉S-3-25人工海水培养基发酵产物中分离得到8个单体化合物,其中化合物1和2为新天然产物,未见有文献数据的报道。化合物1~8均未呈现较强的细胞毒活性。     

酸枣仁总皂苷改善对氯苯丙氨酸诱导失眠大鼠模型学习记忆损伤机制的研究

目的 基于代谢组学技术考察酸枣仁总皂苷（total saponins of Ziziphi Spinosae Semen,SZSS）对失眠大鼠内源性代谢物的影响,并探讨酸枣仁总皂苷改善学习记忆的药效及其作用机制。方法 采用对氯苯丙氨酸（DL-4-chlorophenylalanine,PCPA）诱导大鼠失眠模型,给予地西泮和酸枣仁总皂苷干预后,进行旷场实验及Morris水迷宫实验;采用苏木素-伊红（HE）染色观察海马组织病理变化;采用ELISA测定海马组织5-羟色氨（5-hydroxytryptamine,5-HT）、γ-氨基丁酸（γ-aminobutyric acid,GABA）及NO （nitric oxide,NO）水平。利用UPLC-Orbitrap-MS/MS技术对大鼠海马组织代谢轮廓进行分析,结合多元统计分析筛选差异性代谢物,并基于MetaboAnalyst 5.0数据库进行代谢通路及代谢网络分析,多角度探讨酸枣仁总皂苷改善学习记忆的代谢特征及作用途径。结果 与模型组比较,酸枣仁总皂苷组大鼠学习能力显著提高（P<0.05、0.01）,海马组织病理结构明显改善,海马组织中5-HT、GABA及NO水平显著升高（P<0.05、0.01）;代谢组学共鉴定出18种与学习记忆损伤相关的潜在生物标志物,酸枣仁总皂苷主要通过苯丙氨酸、酪氨酸和色氨酸等7条主要氨基酸代谢通路发挥改善记忆损伤的作用。结论 酸枣仁总皂苷能够通过回调海马组织内源性差异代谢物,并调控相关氨基酸代谢通路,从而改善失眠引起学习记忆减退。     

生物梅里埃肠道细菌药敏试验试剂盒ATB G(-)5临床应用局限性分析

目的：探讨生物梅里埃肠道细菌药敏试验试剂盒ATB G（-）5在临床应用中存在的缺陷,并提出补救（解决）方案。方法以《美国临床和实验室标准化协会（CLSI）M100-S23-表2A肠杆菌科细菌抑菌圈直径和最低抑菌浓度解释标准》（简称CLSI标准）提供的肠杆菌科细菌对应抗菌药物拐点作为标准,对生物梅里埃药敏试验试剂盒ATB G（-）5抗菌药物覆盖情况与各抗菌药物拐点质量浓度进行对比分析。结果试剂盒ATB G（-）5不包括CLSI标准推荐的氯霉素类、磷霉素及硝基呋喃类抗菌药物;试剂盒ATB G（-）5提供的19种抗菌药物拐点均与CLSI标准推荐的抗菌药物拐点不一致或不包含,从而可能导致临床药敏试验报告出现假敏感或假耐药情况。结论以现行CLSI标准作为参考标准,单独使用试剂盒ATB G（-）5进行肠杆菌科细菌药敏试验将产生较大误差,甚至错误,必须用其他药敏试验进行修正方可发出报告。     

Comparative Evaluation of Specific Phytochemical Indicatives in Cirivilvādi Ka?āya Prepared Freshly and at Commercial Scale

Kaṣāya or decoction is an Ayurvedic dosage form, prescribed based on the stage of the disease according to the principles of Ayurveda. This dosage form is traditionally prepared fresh and consumed on the same day but for the sake of convenience; the process of preparation has been modified so that it can be stored with longer shelf life, easy availability and produced in large quantities. There is a need to understand the implications of this modification in terms of chemical changes. This work attempted to check the phytochemical profile of both freshly prepared decoction and commercially available decoction with reference to some analytical parameters like pH, total soluble solids, phenols, alkaloids, potassium and to assess the changes in the thin layer chromatography profiling of the decoction. The results showed that phenols and potassium are found to be two fold higher in freshly prepared decoction, compared to commercially available decoction diluted to dosage in practice (1:4 ratio). However, the total alkaloid content was found to be approximately ten fold higher in commercially available decoction. It was observed that the thin layer chromatography profile of decoctions was extracted into petroleum ether and chloroform was similar and consistent with different batches though the bands in commercially available decoction were slightly more intense compared to freshly prepared decoction. The total soluble solids in commercially available decoction were four times higher than freshly prepared decoction. The study reveals that there are differences in the phytochemical profiles of the freshly prepared decoction and commercially available decoction of the same formulation. However, the significance of these differences can be determined only by further clinical studies. On the other hand, the study lends support to the practice of diluting the commercially available decoction to make it equivalent to freshly prepared decoction.     

Persistent inhibition of Candida albicans biofilm and hyphae growth on titanium by graphene nanocoating

ObjectivesCandida albicanscolonizes biomaterial surfaces and are highly resistant to therapeutics. Graphene nanocoating on titanium compromises initial biofilm formation. However, its sustained antibiofilm potential is unknown. The objective of this study was to investigate the potential of graphene nanocoating to decrease long-term fungal biofilm development and hyphae growth on titanium.MethodsGraphene nanocoating was deposited twice (TiGD) or five times (TiGV) on grade 4 titanium with vacuum assisted technique and characterized with Raman spectroscopy and atomic force microscope. The biofilm formation and hyphae growth of C. albicans was monitored for seven days by CFU, XTT, confocal, mean cell density and scanning electronic microscopy (SEM). Uncoated titanium was the Control. All tests had three independent biological samples and were performed in independent triplicates. Data was analyzed with one- or two-way ANOVA and Tukey's HSD (α = 0.05).ResultsBoth TiGD and TiGV presented less biofilms at all times points compared with Control. The confocal and SEM images revealed few adhered cells on graphene coated samples, absence of hyphae and no features of a mature biofilm architecture. The increase in number of layers of graphene nanocoating did not improve its antibiofilm potential.SignificanceThe graphene nanocoating exerted a long-term persistent inhibitory effect on the biofilm formation on titanium. The fewer cells that were able to attach on graphene coated titanium were scattered and unable to form a mature biofilm with hyphae elements. The findings open opportunities to prevent microbial attachment and proliferation on implantable materials without the use of antibiotics.     

Production of certified reference materials for the sports doping control of the REV‐ERB agonist SR9009

Two human metabolites of the REV‐ERB agonist SR9009, identified by researchers with an interest in sports doping control, have been synthesized and assessed for purity. The synthesis employed was a modification of published procedures for the parent SR9009, careful attention to the purification of intermediates and the final product ensuring materials of the highest purity were available for certification. For each candidate material impurities of related structure were identified and quantified as a relative mass fraction using high performance liquid chromatography–ultraviolet (HPLC–UV) detection and proton nuclear magnetic resonance (¹H NMR) spectroscopy. The quantification of water, occluded solvent, and inorganic residue was assessed using Karl Fischer, ¹H NMR, and thermogravimetric analysis, thereby completing the assessment of all impurities typically characterized by the mass balance approach. Summation and subtraction from 1000 mg/g afforded the mass fraction of the main component, the associated uncertainty ensuring certified reference material status can be applied to the resulting pure substance calibration standards. The availability of these standards to the sports doping control community will facilitate delivery of metrological traceability to the SI unit for mass (kg) to routine testing results and aid method development for the detection and quantification of SR9009 abuse.     

晚期妊娠孕妇B群链球菌定植状况及妊娠结局分析

目的探讨妊娠晚期孕妇B群链球菌（GBS）定植情况、抗菌药物敏感性及妊娠结局。方法纳入2016年1月至2018年12月在东部战区总医院和南京医科大学第一附属医院妇产科定期产检的孕妇，于孕35~37周采用标准方法采集阴道及直肠拭子进行GBS培养，并对阳性标本分离的菌株进行药物敏感性试验。按培养结果分为GBS阳性组和GBS阴性组，阳性组按照产程中是否使用了抗菌药物治疗分为用药组与未用药组，比较不同组别妊娠结局的差异。结果共13 000名孕妇入组，GBS总体定植率为3.65%（475/13 000）。GBS在阴道中定植率为2.33%（303/13 000），在直肠中定植率为1.75%（227/13 000）。通过对直肠标本的采集检测，GBS阳性检出率增加了56.77%（172/303）。GBS每月的定植率有明显波动，3月份和10月份最高（均 P < 0.05）。475份GBS阳性标本对头孢曲松、万古霉素和利奈唑胺的敏感率为100%，对氨苄西林和青霉素的敏感率分别为97.26%和93.47%，而对左氧氟沙星、克林霉素、红霉素、四环素的耐药率较高，分别为30.11%、48.00%、52.21%和88.63%。GBS阳性组胎膜早破、产后出血、产褥期感染、新生儿肺炎、败血症发生率较GBS阴性组明显增高（均 P < 0.01）。产程中预防性使用抗菌药物的GBS阳性孕妇产褥期感染、新生儿感染及新生儿重症监护室入住率明显低于未使用抗菌药物的孕妇（ P < 0.05或 P < 0.01）。 结论妊娠晚期孕妇GBS定植率较低，无明显的季节性，通过补充直肠检测能提高GBS检出率。头孢曲松、氨苄西林和青霉素是目前预防和治疗GBS相关疾病的首选药物。GBS感染会明显增加母婴并发症的发生可能，产程中抗菌药物治疗可以改善母婴的结局。