Objective: To investigate chemical constituents and new antimicrobial agents among essential oils from the rhizomes of Curcuma aeruginosa(C. aeruginosa) Roxb., Curcuma glans K. Larsen & J. Mood and Curcuma cf. xan...Objective: To investigate chemical constituents and new antimicrobial agents among essential oils from the rhizomes of Curcuma aeruginosa(C. aeruginosa) Roxb., Curcuma glans K. Larsen & J. Mood and Curcuma cf. xanthorrhiza Roxb.Methods: The essential oils were obtained by hydro-distillation and analyzed by gas chromatography/mass spectroscopy. Agar-well diffusion assay was used to study the antimicrobial activity and also broth-micro dilution techniques were examined for minimum inhibitory concentration(MIC) against four bacterial strains and yeast.Results: The gas chromatography/mass spectroscopy analysis showed monoterpenes predominantly(88.53%) in the rhizome oil of Curcuma cf. xanthorrhiza. Sesquiterpenes(50.10%) was the most abundant component in the essential oil of C. glans, while monoterpenes(45.55%) and sesquiterpenes(45.81%) were found in C. aeruginosa with a significant amount. The major components of C. aeruginosa were characterized as camphor(29.39%) and germacrone(21.21%). Germacrone(15.76%), b-pinene(9.97%)and camphor(9.96%) were found as major compounds in the rhizome oils of C. glans while a-terpinolene(24.86%) and p-cymen-7-ol(12.17%) were found as major compositions in Curcuma cf. xanthorrhiza. The essential oils were tested against four bacterial strains and yeast. As a result, the rhizome oil of C. aeruginosa exhibited potent activity against Staphylococcus aureus [inhibition zone(21.94 ± 0.24) mm, MIC 125 μg/mL],Bacillus cereus [inhibition zone(20.83 ± 0.36) mm, MIC 125 μg/mL], and Candida albicans [inhibition zone(11.60 ± 0.30) mm, MIC 250 μg/mL].Conclusions: The essential oils from three Curcuma species possessed greater activity against the gram-positive bacteria(Staphylococcus aureus and Bacillus cereus) than gram-negative bacteria(Escherichia coli and Pseudomonas aeruginosa). The results suggest that the essential oils from the fresh rhizome of Curcuma spp. might be a potential source of natural antimicrobial substances.展开更多
Diatoms are the only ecological phytoplankton that require silicic acid for growth.They are also the dominant contributor of ocean's total primary productivity.Generation and circulation with silica walls,which th...Diatoms are the only ecological phytoplankton that require silicic acid for growth.They are also the dominant contributor of ocean's total primary productivity.Generation and circulation with silica walls,which the siliceous organisms form,is an important component of the marine biological pump.It is crucial to the study of the operational mechanisms of biological pump with different sea areas.Moreover,it is the key link to the study of global silicon cycle.This paper introduces the basic mechanism of the formation of diatom silica walls and a new way of researching silicic acid metabolism,namely the 2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)-methoxy)phenyl)oxazole(PDMPO) dyeing method.Under a fluorescence microscope after excitation with bright green fluorescence,it can combine with silicic acid to form a complex into the Si deposition within diatom cells.The advantage of this method is that it can monitor the metabolism of silicate after adding PDMPO.For experimentation and sample collection in each of the specified time points,samples were determinated through the unutilized silicic acid,silica dissoluble intracellular and Si deposition within diatom cells,not only using hot alkaline digestions method but also PDMPO dyeing method.Results showed a good linear relationship between PDMPO fluorescent value and biogenic silica concentration.It was also indicated that PDMPO had no deleterious impact on Skeletonema cf.costatum growth for 34 h and was useful for tracking newly-deposited biogenic silica in diatoms' frustules.展开更多
Several species of the fungal genus Trichoderma establish biological interactions with various micro- and macro-organisms. Some of these interactions are relevant in ecological terms and in biotechnological applicatio...Several species of the fungal genus Trichoderma establish biological interactions with various micro- and macro-organisms. Some of these interactions are relevant in ecological terms and in biotechnological applications, such as biocontrol, where Trichoderma could be considered as an invasive species that colonizes a recipient community. The success of this invasion depends on multiple factors, which can be assayed using experimental communities as study models. Therefore, the aim of this work is to develop a species-specific sequence-characterized amplified region （SCAR） marker to monitor the colonization and growth of T. cf. harzianum when it invades experi- mental communities. For this study, 16 randomly amplified polymorphic DNA （RAPD） primers of 10-mer were used to generate polymorphic patterns, one of which generated a band present only in strains of T. cf. harzianum. This band was cloned, sequenced, and five primers of 20-23 mer were designed. Primer pairs 2F2/2R2 and 2F2/2R3 successfully and specifically amplified fragments of 278 and 448 bp from the T. cf. harzianum BpT10a strain DNA, respectively. Both primer pairs were also tested against the DNA from 14 strains of T. cf. harzianum and several strains of different fungal genera as specificity controls. Only the DNA from the strains of T. cf. hat-zianum was successfully amplified. Moreover, primer pair 2F2/2R2 was assessed by quantitative real-time polymerase chain reaction （PCR） using fungal DNA mixtures and DNA extracted from fungal experimental communities as templates. T. cf. harzianum was detectable even when as few as 100 copies of the SCAR marker were available or even when its population represented only 0.1% of the whole community.展开更多
目的:基于网络药理学和分子对接技术,探讨经验方归元膏的活性成分防治慢性疲劳综合征(CFS)的潜在机制。方法:从TCMSP数据库、BATMAN数据库筛选归元膏的化学成分和靶点,通过GeneCards、OMIM、DisGenet数据库检索收集CFS靶点,将归元膏作...目的:基于网络药理学和分子对接技术,探讨经验方归元膏的活性成分防治慢性疲劳综合征(CFS)的潜在机制。方法:从TCMSP数据库、BATMAN数据库筛选归元膏的化学成分和靶点,通过GeneCards、OMIM、DisGenet数据库检索收集CFS靶点,将归元膏作用靶点和疾病靶点取交集,使用Cytoscape Version 3.9.1制做药物和疾病靶点韦恩图,运用STRING数据库构建蛋白质与蛋白质相互作用的PPI网络作用图,并通过Cytoscape3.8.2软件中的插件CytoNCA筛选关键靶点,并将交集靶点导入David数据库,利用微生信程序进行基因本体(GO)和京都和基因组百科全书(KEGG)富集分析,并通过Cytoscape3.8.2构建归元膏药物–有效成分–靶点和归元膏药物–成分–靶点–通路网络,最后用AutoDock vinna软件进行分子对接验证。结果:检索出归元膏活性成分有111个,其中quercetin,isoflavanone,poriferasta-7,22E-dien-3beta-ol,hancinol,orchinol等为归元膏防治CFS的关键成分,CFS相关靶点507个,其中IL-6 (白细胞介素-6),TNF (肿瘤坏死因子),STAT3 (信号转导及转录激活因子3),JUN,BCL2 (B细胞淋巴瘤/白血病-2),HIF1A (缺氧诱导因子1α亚单位),AKT1 (AKT丝氨酸/苏氨酸激酶1),CASP3 (caspase-3),MMP9 (基质金属蛋白酶9),NFKB1 (核因子kappa B亚基1)为防治CFS的关键靶点,GO富集分析示生物学过程方面可能通过对凋亡过程的负调控、细胞增殖的正调控、蛋白质磷酸化的正调控、内皮细胞增殖的正调控等,分子功能主要包括酶结合、受体结合、蛋白酶结合等;细胞功能主要包含线粒体、蛋白质复合物、细胞内膜结合的细胞器等。KEGG富集分析一共得到186条信号通路,主要与PI3K-Akt通路、卡波西肉瘤相关疱疹病毒感染、丝裂原活化蛋白激酶信号通路等有关。分子对接结果表明归元膏中的核心作用靶点和主要活性成分具备稳定性的结合活性。结论:归元膏将quercetin、poriferasta-7,22E-dien-3beta-ol、orchinol、isoflavanone等关键活性成分与IL6、TNF、STAT3、JUN、BCL2、HIF1A、AKT1和CASP3等靶点结合,通过对PI3K/Akt、MAPK、NF-κB等多条信号通路的调控,调节了CFS患者的免疫反应、细胞凋亡、代谢紊乱等多个生物学过程。通过激活抗凋亡通路、抑制促炎因子的产生和调节能量代谢,验证了该方多靶点多线路的调控,发挥了多靶点、多通路的协同作用,显著改善了CFS的症状。Objective: Based on network pharmacology and molecular docking technology, this study explores the potential mechanisms of active ingredients in the empirical formula Guiyuan Ointment for the prevention and treatment of chronic fatigue syndrome (CFS). Methods: Chemical components and targets of Guiyuan Ointment were screened from the TCMSP database and BATMAN database. CFS targets were collected through GeneCards, OMIM, and DisGenet databases. The intersection of Guiyuan Ointment’s action targets and disease targets was taken. Cytoscape Version 3.9.1 was used to create a Venn diagram of drug and disease targets. The STRING database was employed to construct a protein-protein interaction (PPI) network diagram, and key targets were screened using the CytoNCA plugin in Cytoscape 3.8.2 software. The intersection targets were imported into the David database, and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using the microbioinformatics program. Cytoscape 3.8.2 was used to construct the Guiyuan Ointment drug-effective component-target and drug-component-target-pathway networks. Finally, molecular docking validation was conducted using AutoDock Vina software. Results: A total of 111 active components of Guiyuan Ointment were retrieved, among which quercetin, isoflavanone, poriferasta-7,22E-dien-3beta-ol, hancinol, and orchinol are key components for the prevention and treatment of CFS by Guiyuan Ointment. There are 507 CFS-related targets, with IL-6 (interleukin-6), TNF (tumor necrosis factor), STAT3 (signal transducer and activator of transcription 3), JUN, BCL2 (B-cell lymphoma/leukemia-2), HIF1A (hypoxia-inducible factor 1α subunit), AKT1 (AKT serine/threonine kinase 1), CASP3 (caspase-3), MMP9 (matrix metallopeptidase 9), and NFKB1 (nuclear factor kappa B subunit 1) being key targets for the prevention and treatment of CFS. GO enrichment analysis showed that in terms of biological processes, it may involve negative regulation of apoptosis, positive regulation of cell proliferation, positive regulation of protein phosphorylation, and positive regulation of endothelial cell proliferation, etc. The main molecular functions include enzyme binding, receptor binding, and protease binding, etc. The main cellular functions involve mitochondria, protein complexes, and cell organelles bound to the intracellular membrane, etc. KEGG enrichment analysis yielded a total of 186 signaling pathways, mainly related to the PI3K-Akt pathway, Kaposi’s sarcoma-associated herpesvirus infection, and mitogen-activated protein kinase signaling pathway, etc. The molecular docking results indicated that the core action targets and main active components in Guiyuan Ointment have stable binding activity. Conclusion: Guiyuan Ointment combines key active components such as quercetin, poriferasta-7,22E-dien-3beta-ol, orchinol, and isoflavanone with targets like IL6, TNF, STAT3, JUN, BCL2, HIF1A, AKT1, and CASP3. By regulating multiple signaling pathways such as PI3K/Akt, MAPK, and NF-κB, it modulates various biological processes in CFS patients, including immune response, cell apoptosis, and metabolic disorders. Through activating anti-apoptotic pathways, inhibiting the production of pro-inflammatory factors, and regulating energy metabolism, it verifies the multi-target and multi-pathway regulation of this formula, exerting synergistic effects on multiple targets and pathways, and significantly improving the symptoms of CFS.展开更多
基金supported by the grants from the Biodiversity-Based Economy Development Office Public Organization -National Research Council of Thailand(Grant number.R000012298)Department of Pharmaceutical Sciences,the Faculty of Pharmacy and the Graduate School,Chiang Mai University,Chiang Mai,Thailand
文摘Objective: To investigate chemical constituents and new antimicrobial agents among essential oils from the rhizomes of Curcuma aeruginosa(C. aeruginosa) Roxb., Curcuma glans K. Larsen & J. Mood and Curcuma cf. xanthorrhiza Roxb.Methods: The essential oils were obtained by hydro-distillation and analyzed by gas chromatography/mass spectroscopy. Agar-well diffusion assay was used to study the antimicrobial activity and also broth-micro dilution techniques were examined for minimum inhibitory concentration(MIC) against four bacterial strains and yeast.Results: The gas chromatography/mass spectroscopy analysis showed monoterpenes predominantly(88.53%) in the rhizome oil of Curcuma cf. xanthorrhiza. Sesquiterpenes(50.10%) was the most abundant component in the essential oil of C. glans, while monoterpenes(45.55%) and sesquiterpenes(45.81%) were found in C. aeruginosa with a significant amount. The major components of C. aeruginosa were characterized as camphor(29.39%) and germacrone(21.21%). Germacrone(15.76%), b-pinene(9.97%)and camphor(9.96%) were found as major compounds in the rhizome oils of C. glans while a-terpinolene(24.86%) and p-cymen-7-ol(12.17%) were found as major compositions in Curcuma cf. xanthorrhiza. The essential oils were tested against four bacterial strains and yeast. As a result, the rhizome oil of C. aeruginosa exhibited potent activity against Staphylococcus aureus [inhibition zone(21.94 ± 0.24) mm, MIC 125 μg/mL],Bacillus cereus [inhibition zone(20.83 ± 0.36) mm, MIC 125 μg/mL], and Candida albicans [inhibition zone(11.60 ± 0.30) mm, MIC 250 μg/mL].Conclusions: The essential oils from three Curcuma species possessed greater activity against the gram-positive bacteria(Staphylococcus aureus and Bacillus cereus) than gram-negative bacteria(Escherichia coli and Pseudomonas aeruginosa). The results suggest that the essential oils from the fresh rhizome of Curcuma spp. might be a potential source of natural antimicrobial substances.
基金The National Natural Science Foundation of China primarily supported this studyDr.J.Sun received grants from the National Natural Science Foundation of China(Nos.41276124 and 41676112)+2 种基金Program for Changjiang Scholars and the Science Fund for University Creative Research Groups in Tianjin(No.TD12-5003)Dr.Y.Feng received grants from the National Natural Science Foundation of China(No.41306118)Dr.G.Zhang received grants from the Foundation of Tianjin Key Laboratory of Marine Resources and Chemistry(Tianjin University of Science&Technology),P.R.China(No.201504)
文摘Diatoms are the only ecological phytoplankton that require silicic acid for growth.They are also the dominant contributor of ocean's total primary productivity.Generation and circulation with silica walls,which the siliceous organisms form,is an important component of the marine biological pump.It is crucial to the study of the operational mechanisms of biological pump with different sea areas.Moreover,it is the key link to the study of global silicon cycle.This paper introduces the basic mechanism of the formation of diatom silica walls and a new way of researching silicic acid metabolism,namely the 2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)-methoxy)phenyl)oxazole(PDMPO) dyeing method.Under a fluorescence microscope after excitation with bright green fluorescence,it can combine with silicic acid to form a complex into the Si deposition within diatom cells.The advantage of this method is that it can monitor the metabolism of silicate after adding PDMPO.For experimentation and sample collection in each of the specified time points,samples were determinated through the unutilized silicic acid,silica dissoluble intracellular and Si deposition within diatom cells,not only using hot alkaline digestions method but also PDMPO dyeing method.Results showed a good linear relationship between PDMPO fluorescent value and biogenic silica concentration.It was also indicated that PDMPO had no deleterious impact on Skeletonema cf.costatum growth for 34 h and was useful for tracking newly-deposited biogenic silica in diatoms' frustules.
基金Project supported by the Chilean National Fund for Scientific and Technological Development(FONDECYT)(No.1120508)
文摘Several species of the fungal genus Trichoderma establish biological interactions with various micro- and macro-organisms. Some of these interactions are relevant in ecological terms and in biotechnological applications, such as biocontrol, where Trichoderma could be considered as an invasive species that colonizes a recipient community. The success of this invasion depends on multiple factors, which can be assayed using experimental communities as study models. Therefore, the aim of this work is to develop a species-specific sequence-characterized amplified region （SCAR） marker to monitor the colonization and growth of T. cf. harzianum when it invades experi- mental communities. For this study, 16 randomly amplified polymorphic DNA （RAPD） primers of 10-mer were used to generate polymorphic patterns, one of which generated a band present only in strains of T. cf. harzianum. This band was cloned, sequenced, and five primers of 20-23 mer were designed. Primer pairs 2F2/2R2 and 2F2/2R3 successfully and specifically amplified fragments of 278 and 448 bp from the T. cf. harzianum BpT10a strain DNA, respectively. Both primer pairs were also tested against the DNA from 14 strains of T. cf. harzianum and several strains of different fungal genera as specificity controls. Only the DNA from the strains of T. cf. hat-zianum was successfully amplified. Moreover, primer pair 2F2/2R2 was assessed by quantitative real-time polymerase chain reaction （PCR） using fungal DNA mixtures and DNA extracted from fungal experimental communities as templates. T. cf. harzianum was detectable even when as few as 100 copies of the SCAR marker were available or even when its population represented only 0.1% of the whole community.
文摘目的:基于网络药理学和分子对接技术,探讨经验方归元膏的活性成分防治慢性疲劳综合征(CFS)的潜在机制。方法:从TCMSP数据库、BATMAN数据库筛选归元膏的化学成分和靶点,通过GeneCards、OMIM、DisGenet数据库检索收集CFS靶点,将归元膏作用靶点和疾病靶点取交集,使用Cytoscape Version 3.9.1制做药物和疾病靶点韦恩图,运用STRING数据库构建蛋白质与蛋白质相互作用的PPI网络作用图,并通过Cytoscape3.8.2软件中的插件CytoNCA筛选关键靶点,并将交集靶点导入David数据库,利用微生信程序进行基因本体(GO)和京都和基因组百科全书(KEGG)富集分析,并通过Cytoscape3.8.2构建归元膏药物–有效成分–靶点和归元膏药物–成分–靶点–通路网络,最后用AutoDock vinna软件进行分子对接验证。结果:检索出归元膏活性成分有111个,其中quercetin,isoflavanone,poriferasta-7,22E-dien-3beta-ol,hancinol,orchinol等为归元膏防治CFS的关键成分,CFS相关靶点507个,其中IL-6 (白细胞介素-6),TNF (肿瘤坏死因子),STAT3 (信号转导及转录激活因子3),JUN,BCL2 (B细胞淋巴瘤/白血病-2),HIF1A (缺氧诱导因子1α亚单位),AKT1 (AKT丝氨酸/苏氨酸激酶1),CASP3 (caspase-3),MMP9 (基质金属蛋白酶9),NFKB1 (核因子kappa B亚基1)为防治CFS的关键靶点,GO富集分析示生物学过程方面可能通过对凋亡过程的负调控、细胞增殖的正调控、蛋白质磷酸化的正调控、内皮细胞增殖的正调控等,分子功能主要包括酶结合、受体结合、蛋白酶结合等;细胞功能主要包含线粒体、蛋白质复合物、细胞内膜结合的细胞器等。KEGG富集分析一共得到186条信号通路,主要与PI3K-Akt通路、卡波西肉瘤相关疱疹病毒感染、丝裂原活化蛋白激酶信号通路等有关。分子对接结果表明归元膏中的核心作用靶点和主要活性成分具备稳定性的结合活性。结论:归元膏将quercetin、poriferasta-7,22E-dien-3beta-ol、orchinol、isoflavanone等关键活性成分与IL6、TNF、STAT3、JUN、BCL2、HIF1A、AKT1和CASP3等靶点结合,通过对PI3K/Akt、MAPK、NF-κB等多条信号通路的调控,调节了CFS患者的免疫反应、细胞凋亡、代谢紊乱等多个生物学过程。通过激活抗凋亡通路、抑制促炎因子的产生和调节能量代谢,验证了该方多靶点多线路的调控,发挥了多靶点、多通路的协同作用,显著改善了CFS的症状。Objective: Based on network pharmacology and molecular docking technology, this study explores the potential mechanisms of active ingredients in the empirical formula Guiyuan Ointment for the prevention and treatment of chronic fatigue syndrome (CFS). Methods: Chemical components and targets of Guiyuan Ointment were screened from the TCMSP database and BATMAN database. CFS targets were collected through GeneCards, OMIM, and DisGenet databases. The intersection of Guiyuan Ointment’s action targets and disease targets was taken. Cytoscape Version 3.9.1 was used to create a Venn diagram of drug and disease targets. The STRING database was employed to construct a protein-protein interaction (PPI) network diagram, and key targets were screened using the CytoNCA plugin in Cytoscape 3.8.2 software. The intersection targets were imported into the David database, and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using the microbioinformatics program. Cytoscape 3.8.2 was used to construct the Guiyuan Ointment drug-effective component-target and drug-component-target-pathway networks. Finally, molecular docking validation was conducted using AutoDock Vina software. Results: A total of 111 active components of Guiyuan Ointment were retrieved, among which quercetin, isoflavanone, poriferasta-7,22E-dien-3beta-ol, hancinol, and orchinol are key components for the prevention and treatment of CFS by Guiyuan Ointment. There are 507 CFS-related targets, with IL-6 (interleukin-6), TNF (tumor necrosis factor), STAT3 (signal transducer and activator of transcription 3), JUN, BCL2 (B-cell lymphoma/leukemia-2), HIF1A (hypoxia-inducible factor 1α subunit), AKT1 (AKT serine/threonine kinase 1), CASP3 (caspase-3), MMP9 (matrix metallopeptidase 9), and NFKB1 (nuclear factor kappa B subunit 1) being key targets for the prevention and treatment of CFS. GO enrichment analysis showed that in terms of biological processes, it may involve negative regulation of apoptosis, positive regulation of cell proliferation, positive regulation of protein phosphorylation, and positive regulation of endothelial cell proliferation, etc. The main molecular functions include enzyme binding, receptor binding, and protease binding, etc. The main cellular functions involve mitochondria, protein complexes, and cell organelles bound to the intracellular membrane, etc. KEGG enrichment analysis yielded a total of 186 signaling pathways, mainly related to the PI3K-Akt pathway, Kaposi’s sarcoma-associated herpesvirus infection, and mitogen-activated protein kinase signaling pathway, etc. The molecular docking results indicated that the core action targets and main active components in Guiyuan Ointment have stable binding activity. Conclusion: Guiyuan Ointment combines key active components such as quercetin, poriferasta-7,22E-dien-3beta-ol, orchinol, and isoflavanone with targets like IL6, TNF, STAT3, JUN, BCL2, HIF1A, AKT1, and CASP3. By regulating multiple signaling pathways such as PI3K/Akt, MAPK, and NF-κB, it modulates various biological processes in CFS patients, including immune response, cell apoptosis, and metabolic disorders. Through activating anti-apoptotic pathways, inhibiting the production of pro-inflammatory factors, and regulating energy metabolism, it verifies the multi-target and multi-pathway regulation of this formula, exerting synergistic effects on multiple targets and pathways, and significantly improving the symptoms of CFS.
