基于网络药理学与分子对接技术的清瘟护肺颗粒防治新型冠状病毒肺炎(COVID-19)的潜在药效物质研究

本文通过网络药理学和分子对接技术探讨清瘟护肺颗粒防治新型冠状病毒肺炎(COVID-19)的潜在药效物质。首先,通过TCMSP数据库,BATMAN-TCM数据库及TCMIP数据库检索清瘟护肺颗粒中十六味药的化学成分及作用靶点,利用GeneCards和OMIM数据库检索COVID-19的相关疾病靶点。然后,通过venny2.1.0获取清瘟护肺颗粒防治COVID-19的潜在靶点,利用R语言对潜在靶点进行GO功能和KEGG通路富集分析,并结合文献对富集所得通路进行分析。最后,利用Cytoscape3.7.1软件构建网络图,采用AutoDock4.2.1软件评价清瘟护肺颗粒中潜在药效成分和新型冠状病毒SARS-CoV-23CL水解酶、血管紧张素转化酶II(ACE2)和RNA依赖的RNA聚合酶(RdRp)的结合作用。网络药理学得到清瘟护肺颗粒防治COVID-19的473个活性成分和123个靶点,KEGG结果及文献分析预测到清瘟护肺颗粒可通过调控MAPK、小细胞肺癌、肺结核、PI3K-AKT等多条信号通路而发挥作用,分子对接结果显示清瘟护肺颗粒中潜在药效成分和SARS-CoV-23CL水解酶、ACE2及RdRp具有良好的亲和性。本研究较为全面揭示了清瘟护肺颗粒治疗COVID-19“多成分、多靶点、多通路”的特点,为深入探讨清瘟护肺颗粒治疗COVID-19的作用机制提供参考依据。     

Main antimicrobial components of Tinospora capillipes, and their mode of action against Staphylococcus aureus

In this investigation, the antibacterial modes of action of Radix Tinosporae, its major single components, and nine antibiotics with different targets or modes-of-action on Staphylococcus aureus were studied. Metabolic profiles of cultures treated with different medicines were acquired by HPLC/ESI-MS. After HPLC-MS data pretreatment, those profiles acquired were reduced into several MS vectors. Then statistical processing by principal components analysis was carried out upon those vectors, two conclusions could be drawn: (1) the antibacterial mode of action of Radix Tinosporae is similar to that of rifampicin and norfloxacin, which act on nucleic acid; (2) its active components playing main antimicrobial roles on Staphylococcus aureus might be alkaloids, such as palmatine and jatrorrhizine.     

灰胸竹鸡的生物学特性及经济价值

本文从形态特征、生活习性、繁殖和血液生理生化特点等方面介绍了灰胸竹鸡生物学特性和经济价值,对灰胸竹鸡的人工驯养与繁殖,以及保护性开发具有重要意义。     

花椰菜ACC氧化酶基因的克隆及其RNAi对内源基因表达的抑制作用

根据亲缘关系较近的几种作物ACC氧化酶氨基酸序列,设计一对简并引物,从花椰菜(Brassica oleracea var．botrytis)基因组中获得长1202bp的候选片段。序列分析表明该基因含有3个外显子(Exon)、2个内含子(Intron),编码区序列长756bp,编码252个氨基酸。同源性分析发现其与青花菜(Brassica oleracea var．Italica)上已发表mRNA序列同源率为99％(GenBank序列号：X81629),推断该基因为花椰菜ACC氧化酶基因,命名为BoACO(GenBank登录号：AY676466)。在此基础上,用BP克隆的方法构建BoACO的RNA干涉(RNAi)载体pHBACO,对花椰菜进行遗传转化,获得卡那霉素抗性转化植株5棵,分子检测证实外源片段成功导入其中3棵花椰菜基因组中。Northern杂交分析显示：转基因植株内源ACO基因转录的mRNA被降解。ACC氧化酶活性分析进一步表明,外源基因的导人大大地降低了ACC氧化酶活性。     

灰胸竹鸡消化系统形态解剖初探

观察和测定了13只灰胸竹鸡消化系统的解剖参数,结果表明:雌性和雄性体重分别为227.39 g和216.79 g,体长分别为237.22 mm和230.00 mm;腺胃粘膜表面具有40～50枚圆形乳头,肌胃较发达;雌性和雄性的食道分别长约93.96 mm和99.75 mm,肠道总长分别为683.83 mm和672.95 mm,肝脏分别重4.87 g和5.20 g,胰脏分别重0.38 g和0.45 g.此研究为进行竹鸡的人工驯养和生物学研究提供解剖学资料.     

相对定量和绝对定量-以CsSAMDC基因表达分析为例

以无芒隐子草干旱诱导的cDNA文库中获得的SAMDC序列为基础,应用实时荧光定量RT-PCR技术同时结合相对定量和绝对定量方法,以分析无芒隐子草SAMDC基因在不同组织、不同胁迫梯度下的表达为案例,详细介绍并比较相对定量和绝对定量在基因表达方面的应用.分别采集无芒隐子草幼苗自然干旱0,2,4,6,8,10d以及恢复浇水1,4d的叶片和根系材料,相对定量采用2-ΔΔCT.法;以相应基因的质粒扩增产物为标准品制作标准曲线,对定量PCR的引物浓度进行了优化,而后筛选最稳定的内参基因,以内参基因获得的标准化因子对绝对定量数据进行标准化.相对定量2-ΔΔCT法结果显示CsSAMDC基因在干旱胁迫8d后,叶片中的表达量为胁迫前的0.62±0.13倍,根中的表达量为胁迫前的5.93±0.71倍.绝对定量表明,CsSAMDC基因只在胁迫第8d和第10d的根组织中检测到表达量显著增大,胁迫第4～6d和恢复浇水后1～4d,CsSAMDC基因在叶和根组织中的表达较对照降低.相对定量和绝对定量在DsSAMDC基因表达研究中显示相对一致的趋势.     

Effects of salt and drought stress on alkaloid production in endophyte-infected drunken horse grass (Achnatherum inebrians)

An evaluation was made of the influence of salt stress (0, 100, 200 or 300 mmol/l) and drought stress (60% Water-Holding Capacity, 40% WHC or 20% WHC) on the levels of ergot alkaloids (ergonovine and ergine) formed in drunken horse grass (Achnatherum inebrians) grown under greenhouse conditions. Samples were collected and randomized, both at the beginning of the treatment and after treatment, and the alkaloids were extracted and analyzed by reverse-phase HPLC. Higher alkaloid levels were recorded for plants under salt or drought stress, with levels of ergonovine being higher than those of ergine. Concentrations of both alkaloids increased over the plant growing period.     

Validate antibacterial mode and find main bioactive components of traditional Chinese medicine Aquilegia oxysepala

Traditional Chinese medicines have been used for thousands of years and are still being used as one of the regular treatments for many diseases. However, their mechanisms were still unknown. In this investigation, a possible procedure combining metabonomics and principal component analysis to investigate antibacterial modes of action and find main antimicrobial component in traditional Chinese medicine, Aquilegia oxysepala, is developed. Metabolic profiles of Staphylococcus aureus treated with nine antibiotics of known modes of action and with A. oxysepala were acquired by HPLC/DAD/ESI-MS. After statistical processing by principal components analysis on metabolic profiles, two conclusions could be drawn: (1) the target of A. oxysepala may be similar to that of lincolmensin, erythromycin, chloromycetin, streptomycin, and acheomycin, whose targets are protein; (2) its bioactive component playing main antimicrobial roles on S. aureus may be maguoflorine.     

保水功能菌枯草芽胞杆菌LX-1固态发酵的工艺优化

γ-聚谷氨酸(γ-PGA)对土壤保水和提高植物抗逆性具有明显作用,为了利用农产品加工下脚料发酵生产含有γ-PGA的保水功能肥料,对福建省微生物研究所环境微生物研究室分离到的产γ-PGA的枯草芽胞杆菌(Bacillus subtilis)LX-1的固态发酵培养基配比进行优化。实验以发酵产物的荧光素双醋酸酯酶(FDA)酶活和增比黏度为指标,在通过单因素实验选定固态发酵料的碳源、氮源、无机盐溶液的最佳配比以及无机盐较佳添加量的基础上,通过拟水平均匀设计U₇* (7⁴试验,得出菌株LX-1固态发酵的最佳培养基配方：V豆粕V麦麸V无机盐溶液=644,其中无机盐溶液的最佳配方为K₂HPO₄·3H₂O 5.24 g/L、MgSO₄·7H₂O 0.78 g/L、NaCl 11.67 g/L。为利用农产品加工下脚料发酵生产含γ-PGA保水肥料提供参考。     

Invasiveness of mouse embryos to human ovarian cancer cells HO8910PM and the role of MMP-9

ABSTRACT: Background Our previous work found that mouse embryos could invade malignant cancer cells. In the process of implantation, embryo trophoblast cells express matrix metalloproteinases and the invasive ability of trophoblast cells is proportional to matrix metalloproteinase-9 protein expression. So the purpose of this study is to observe the effects of mouse embryos on human ovarian cancer cells in the co-culture environment in vitro and explore the possible mechanism of matrix metalloproteinase-9. Methods Several groups of human ovarian cancer cells HO8910PM were co-cultured with mouse embryos for different time duration, after which the effects of mouse embryos on morphology and growth behavior of HO8910PM were observed under the light microscope real-time or by H.E staining. Apoptosis was detected under laser confocal microscope by Annexin V-EGFP/PI staining in situ. Invasion ability of tumor cells was studied by transwell experiments. After matrix metalloproteinase 9 (MMP -9) activity was inhibited by MMP-9 Inhibitor I, the interaction between mouse embryos and human ovarian cancer cells HO8910PM was observed. Results Mouse embryos were able to invade co-cultured human ovarian cancer cell layer which extended in the bottom of the culture dish, and gradually pushed away tumor cells to form their own growth space. The number of apoptosis tumor cells surrounding the embryo increased under laser confocal microscope. After co-cultured with mouse embryos, tumor cells invasive ability was lowered compared with the control group. After MMP-9 activity was inhibited, the interaction between mouse embryos and HO8910PM cells had no significant difference compared with the normal MMP-9 activity group. Conclusion Mouse embryos were able to invade human ovarian cancer cells in vitro and form their own growth space, promote apoptosis of human ovarian cancer cells and lower their invasive ability. The mouse embryo was still able to invade human ovarian cancer cells after MMP-9 activity was inhibited.