1.Analysis of chemical constituents in ethyl acetate extract of Taxilli Herba by UPLC-Q-Exactive-MS and screening of potential xanthine oxidase inhibitors.
Yuan LIANG ; Li LI ; Yi CAI ; Li-Ba XU ; Feng-Feng XIE ; Da-Long LIANG ; Fu-Lai CHEN
China Journal of Chinese Materia Medica 2022;47(4):972-979
The present study analyzed and identified the chemical constituents from ethyl acetate(EA) extract of Taxilli Herba with UPLC-Q-Exactive-MS and screened active xanthine oxidase(XO) inhibitors with HPLC. The analysis was performed on an Hypersil GOLD C_(18) reversed-phase column(2.1 mm×50 mm, 1.9 μm), with the mobile phase of water containing 1% formic acid(A) and methanol(B) under gradient elution, the flow rate of 0.3 mL·min~(-1), and the injection volume of 5 μL. ESI source was used for MS and the compounds were collected in positive and negative ion modes. Xcalibur 4.1 was used to analyze the retention time, accurate relative molecular weight, and fragmentation of the compounds. The inhibitory activity of some known compounds on XO was screened by HPLC. Thirty chemical constituents were identified, including phenolic acids and flavonoids by experimental data combined with information of standards, data reported previously, and databases, such as MzCloud and ChemSpider. The activities of 10 chemical components were screened. Gallic acid and naringenin chalcone had strong inhibitory activities on XO with IC_(50) of 57 μg·mL~(-1) and 108 μg·mL~(-1). UPLC-Q-Exactive-MS allows the accurate, rapid, and comprehensive identification of main chemical constituents from Taxilli Herba. Gallic acid and naringenin chalcone may be the active components of XO inhibitors.
Acetates
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Chromatography, High Pressure Liquid
;
Drugs, Chinese Herbal/chemistry*
;
Tandem Mass Spectrometry
;
Xanthine Oxidase
2.Chemical constituents from ethyl acetate extraction of Prunus mume.
Can-Can WANG ; Wei ZHANG ; De-Ling WU ; Feng-Qing XU ; Jin-Song LIU ; Jin-Wei QIAO
China Journal of Chinese Materia Medica 2020;45(2):347-351
Eight compounds were isolated from the ethyl acetate extraction of Prunus mume by column chromatography. On the basis of physicochemical properties and spectrum analysis, these compounds were identified as isoquercitrin-6″-O-benzoate(1), pinoresinol(2), naringin(3), ethyl-β-D-glucopyranoside(4), astragalin(5), quercetin(6), hypericin(7), and rutin(8). Among them, compound 1 was a new natural product, and compounds 2-5 were isolated from this plant for the first time. In vitro study, compounds 1, 3, 5-8 could significantly increase the cell survival ratio.
Acetates
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Phytochemicals/analysis*
;
Plant Extracts/chemistry*
;
Prunus/chemistry*
;
Solvents
3.Chemical constituents from ethyl acetate-soluble extraction of Valeriana jatamansi.
Sheng LIN ; Peng FU ; Yun-Heng SHEN ; Ji YE ; Zhong-Xiao ZHANG ; Xian-Wen YANG ; Hui-Liang LI ; Run-Hui LIU ; Xi-Ke XU ; Wei-Dong ZHANG
China Journal of Chinese Materia Medica 2018;43(1):100-108
Application of a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, macroporous adsorbent resin, and reversed-phase HPLC, led to the isolation of 173 compounds including irdidoids, monoterpenes, sesquiterpenes, triterpenes, lignans, flavonoids, and simple aromatic derivatives from the ethyl acetate-soluble fraction of the whole plants of Valeriana jatamansi(Valerianaceae), and their structures were elucidated by spectroscopic methods including 1D, 2D NMR UV, IR, and MS techniques. Among them, 77 compounds were new. In previous reports, we have described the isolation, structure elucidation, and bioactivities of 68 new and 25 known compounds. As a consequence, we herein reported the isolation and structure elucidation of the remaining 9 new and 71 known compounds, the structure revision of valeriotriate A(8a), as well as cytotoxicity of some compounds.
Acetates
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Chromatography, High Pressure Liquid
;
Flavonoids
;
analysis
;
Iridoids
;
analysis
;
Lignans
;
analysis
;
Molecular Structure
;
Monoterpenes
;
analysis
;
Phytochemicals
;
analysis
;
Plant Extracts
;
chemistry
;
Sesquiterpenes
;
analysis
;
Triterpenes
;
analysis
;
Valerian
;
chemistry
4.An oriental melon 9-lipoxygenase gene CmLOX09 response to stresses, hormones, and signal substances.
Li-Jun JU ; Chong ZHANG ; Jing-Jing LIAO ; Yue-Peng LI ; Hong-Yan QI
Journal of Zhejiang University. Science. B 2018;19(8):596-609
In plants, lipoxygenases (LOXs) play a crucial role in biotic and abiotic stresses. In our previous study, five 13-LOX genes of oriental melon were regulated by abiotic stress but it is unclear whether the 9-LOX is involved in biotic and abiotic stresses. The promoter analysis revealed that CmLOX09 (type of 9-LOX) has hormone elements, signal substances, and stress elements. We analyzed the expression of CmLOX09 and its downstream genes-CmHPL and CmAOS-in the leaves of four-leaf stage seedlings of the oriental melon cultivar "Yumeiren" under wound, hormone, and signal substances. CmLOX09, CmHPL, and CmAOS were all induced by wounding. CmLOX09 was induced by auxin (indole acetic acid, IAA) and gibberellins (GA3); however, CmHPL and CmAOS showed differential responses to IAA and GA3. CmLOX09, CmHPL, and CmAOS were all induced by hydrogen peroxide (H2O2) and methyl jasmonate (MeJA), while being inhibited by abscisic acid (ABA) and salicylic acid (SA). CmLOX09, CmHPL, and CmAOS were all induced by the powdery mildew pathogen Podosphaera xanthii. The content of 2-hexynol and 2-hexenal in leaves after MeJA treatment was significantly higher than that in the control. After infection with P. xanthii, the diseased leaves of the oriental melon were divided into four levels-levels 1, 2, 3, and 4. The content of jasmonic acid (JA) in the leaves of levels 1 and 3 was significantly higher than that in the level 0 leaves. In summary, the results suggested that CmLOX09 might play a positive role in the response to MeJA through the hydroperoxide lyase (HPL) pathway to produce C6 alcohols and aldehydes, and in the response to P. xanthii through the allene oxide synthase (AOS) pathway to form JA.
Abscisic Acid
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Acetates/chemistry*
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Aldehyde-Lyases/metabolism*
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Aldehydes/chemistry*
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Cucurbitaceae/genetics*
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Cyclopentanes/chemistry*
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Cytochrome P-450 Enzyme System/metabolism*
;
Gene Expression Profiling
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Gene Expression Regulation, Plant
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Hormones/metabolism*
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Hydrogen Peroxide/metabolism*
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Intramolecular Oxidoreductases/metabolism*
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Lipoxygenase/metabolism*
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Oxylipins/chemistry*
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Plant Leaves/genetics*
;
Plant Proteins/metabolism*
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Promoter Regions, Genetic
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Salicylic Acid/chemistry*
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Seedlings/metabolism*
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Signal Transduction
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Stress, Physiological
;
Transgenes
5.Anti-inflammatory and membrane stabilizing properties of methyl jasmonate in rats.
Solomon UMUKORO ; Akinyinka Oladipo ALABI ; Anthony Taghogho EDUVIERE ; Abayomi Mayowa AJAYI ; Oluwafemi Gabriel OLUWOLE
Chinese Journal of Natural Medicines (English Ed.) 2017;15(3):202-209
The present investigation was carried out to evaluate anti-inflammatory and membrane stabilizing properties of methyl jasmonate (MJ) in experimental rat models of acute and chronic inflammation. The effects of MJ on acute inflammation were assessed using carrageenan-induced rat's paw edema model. The granuloma air pouch model was employed to evaluate the effects of MJ on chronic inflammation produced by carrageenan in rats. The number of white blood cells (WBC) in pouch exudates was estimated using light microscopy. The levels of biomarkers of oxidative stress, such as malondialdehyde (MDA), glutathione (GSH) and activity of antioxidant enzymes in the exudates, were determined using spectrophotometry. The membrane stabilizing property of MJ was assessed based on inhibition of hemolysis of rat red blood cells (RBC) exposed to hypotonic medium. Our results indicated that MJ (25-100 mg·kg, i.p.) produced significant anti-inflammatory activity in carrageenan-induced paw edema in rats (P < 0.05). MJ reduced the volume of pouch exudates and the number of WBC in carrageenan-induced granulomatous inflammation. It also exhibited potent antioxidant and membrane stabilizing activities. In conclusion, these findings suggest the therapeutic potentials of methyl jasmonate in disease conditions associated with inflammation and its anti-inflammatory activity may be related to its antioxidant and membrane stabilizing activities.
Acetates
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administration & dosage
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Animals
;
Anti-Inflammatory Agents
;
administration & dosage
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Cell Membrane
;
chemistry
;
drug effects
;
immunology
;
Cyclopentanes
;
administration & dosage
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Disease Models, Animal
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Edema
;
drug therapy
;
immunology
;
Erythrocytes
;
chemistry
;
drug effects
;
Glutathione
;
immunology
;
Humans
;
Male
;
Malondialdehyde
;
immunology
;
Oxylipins
;
administration & dosage
;
Plant Extracts
;
administration & dosage
;
Rats
;
Rats, Wistar
6.Synergistic Anti-bacterial Effects of Phellinus baumii Ethyl Acetate Extracts and beta-Lactam Antimicrobial Agents Against Methicillin-Resistant Staphylococcus aureus.
Seung Bok HONG ; Man Hee RHEE ; Bong Sik YUN ; Young Hoon LIM ; Hyung Geun SONG ; Kyeong Seob SHIN
Annals of Laboratory Medicine 2016;36(2):111-116
BACKGROUND: The development of new drugs or alternative therapies effective against methicillin-resistant Staphylococcus aureus (MRSA) is of great importance, and various natural anti-MRSA products are good candidates for combination therapies. We evaluated the antibacterial activities of a Phellinus baumii ethyl acetate extract (PBEAE) and its synergistic effects with beta-lactams against MRSA. METHODS: The broth microdilution method was used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the PBEAE. The PBEAE synergistic effects were determined by evaluating the MICs of anti-staphylococcal antibiotic mixtures, with or without PBEAE. Anti-MRSA synergistic bactericidal effects of the PBEAE and beta-lactams were assessed by time-killing assay. An ELISA was used to determine the effect of the PBEAE on penicillin binding protein (PBP)2a production. RESULTS: The MICs and MBCs of PBEAE against MRSA were 256-512 and 1,024-2,048 microg/mL, respectively. The PBEAE significantly reduced MICs of all beta-lactams tested, including oxacillin, cefazolin, cefepime, and penicillin. However, the PBEAE had little or no effect on the activity of non-beta-lactams. Time-killing assays showed that the synergistic effects of two beta-lactams (oxacillin and cefazolin) with the PBEAE were bactericidal in nature (Deltalog10 colony forming unit/mL at 24 hr: 2.34-2.87 and 2.10-3.04, respectively). The PBEAE induced a dose-dependent decrease in PBP2a production by MRSA, suggesting that the inhibition of PBP2a production was a major synergistic mechanism between the beta-lactams and the PBEAE. CONCLUSIONS: PBEAE can enhance the efficacy of beta-lactams for combined therapy in patients infected with MRSA.
Acetates/chemistry
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Agaricales/*chemistry/metabolism
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Anti-Infective Agents/chemistry/*pharmacology
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Drug Synergism
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Enzyme-Linked Immunosorbent Assay
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Methicillin-Resistant Staphylococcus aureus/*drug effects/metabolism
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Microbial Sensitivity Tests
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Penicillin-Binding Proteins/analysis/metabolism
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Plant Extracts/chemistry/*pharmacology
;
beta-Lactams/*pharmacology
7.Effect of glyceryl triacetate on properties of PLA/PBAT blends.
Nan YANG ; Xiyuan WANG ; Yunxuan WENG ; Yujuan JIN ; Min ZHANG
Chinese Journal of Biotechnology 2016;32(6):839-847
Poly lactic acid (PLA)/Poly (butyleneadipate-co-terephthalate)(PBAT) and glyceryl triacetate (GTA) blend were prepared by torque rheometer, and the effect of GTA on thermodynamical performance, mechanical properties and microstructure of PLA/PBAT composites were studied using differential scanning calorimeter(DSC), dynamic mechanical analysis(DMA), universal testing machine, impact testing machine and scanning electron microscope(SEM). After adding GTA, Tg values of the two phases gradually became closer, blends cold crystallization temperature and melting temperature decreased. When with 3 phr GTA, the dispersed phase particle size of PLA/PBAT blend decreased. Mechanics performance test showed that the elongation at break and impact strength of the PLA/PBAT blend was greatly increased with 3 phr GTA, and the elongation at break increased 2.6 times, improved from 17.7% to 64.1%.
Acetates
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chemistry
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Calorimetry, Differential Scanning
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Crystallization
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Lactic Acid
;
Polyesters
;
chemistry
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Polymers
;
Temperature
8.HPLC-based activity profiling of anti-hepatocellular carcinoma constituents from the Tibetan medicine, Caragana tibetica.
Ping SONG ; Qiang WANG ; Jing-Nan LV ; Chan XU ; Qin-Xiong LIN ; Xin-Hua MA ; Mi HUANG ; Xin-Zhou YANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(3):450-455
During the screening of a traditional Chinese folk herb library against HepG2 and Hep3B cell lines, the EtOAc extract from the Tibetan medicine, Caragana tibetica (CT-EtOAc) exhibited potential anti-hepatocellular carcinoma (anti-HCC) activity. HPLC-based activity profiling was performed for targeted identification of anti-HCC activity from CT-EtOAc by MS-directed purification method. CT-EtOAc was separated by time-based fractionation for further anti-HCC bioassay by a semipreparative HPLC column (150 mm × 10 mm i.d., 5 μm) with a single injection of 5 mg. Bioassay-guided and ESIMS-directed large scale purification was performed with a single injection of 400 mg of CT-EtOAc by peak-based fractionation. A 1.4-mm heavy wall micro NMR tube with z-gradient was used to measure one and two dimensional NMR spectra for the minor or trace amounts of components of the extract. Two active compounds could be elucidated as naringenin chalcone (CT-1) and 3-hydroxy-8, 9-dimethoxypterocarpan (CT-2) relevant to anti-HCC effects for the EtOAc extract of C. tibetica rapidly and unambiguously by this protocol.
Acetates
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pharmacology
;
Antineoplastic Agents
;
chemistry
;
pharmacology
;
Caragana
;
chemistry
;
Carcinoma, Hepatocellular
;
drug therapy
;
Cell Line, Tumor
;
Chalcones
;
pharmacology
;
Chromatography, High Pressure Liquid
;
Hep G2 Cells
;
Humans
;
Liver Neoplasms
;
drug therapy
;
Medicine, Tibetan Traditional
;
Plant Extracts
;
chemistry
;
pharmacology
;
Plant Roots
;
chemistry
9.Expression analyses of BcUGT3 and BcUGT6, and their in vitro expression in Escherichia coli.
Yun-Wen TAO ; Jie-Sen XU ; Jing SUN ; Jian-He WEI ; Juan LIU ; Chun SUI
China Journal of Chinese Materia Medica 2014;39(2):185-191
The tissue-specific and MeJA-induced transcriptional levels of BcUGT3 and BcUGT6 in Bupleurum chinense were analyzed in the present study. The transcriptional levels of BcUGT3 in root, leaf, flower and fruit were similar and they all were higher than those in stem. The transcriptional level of BcUGT6 was the highest in leaf and the lowest in flower among in all tested tissues. With non-treated adventitious roots as control, BcUGT6's transcriptional levels were elevated to nearly 2 folds for 2 h, 8 h, 24 h, 2 d and 4 d in MeJA-treated adventitious roots of B. chinense. It showed that the transcriptional level of BcUGT6 was slightly affected by MeJA. While, BcUGT3's transcriptional levels were gradually elevated, and till 4 d after MeJA treatment, the expression level was about 7 folds than that of non-treated control. Using pET-28a (+), the expressions of two genes was investigated. Induced by IPTG, the target proteins were expressed in E. coli and then purified. All the results obtained in the present study will be helpful for follow-up bio-function analysis of BcUGT3 and BcUGT6.
Acetates
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pharmacology
;
Bupleurum
;
cytology
;
enzymology
;
genetics
;
Cell Membrane
;
metabolism
;
Cyclopentanes
;
pharmacology
;
Escherichia coli
;
genetics
;
Gene Expression
;
Gene Expression Regulation, Plant
;
drug effects
;
Hexosyltransferases
;
chemistry
;
genetics
;
isolation & purification
;
metabolism
;
Intracellular Space
;
metabolism
;
Oxylipins
;
pharmacology
;
Protein Sorting Signals
;
Protein Structure, Secondary
;
Protein Transport
;
Sequence Analysis
;
Transcription, Genetic
;
drug effects
10.Technological parameter optimization method for washing of coarse bear gall powder extracts based on NIR spectroscopy.
Wen-Long LI ; Shao-Yong LIU ; Dong-Sheng XUE ; Zhi-Wei CHENG ; Hai-Bin QU
China Journal of Chinese Materia Medica 2014;39(1):75-79
During the washing process of coarse bear gall powder extracts, it is necessary to adjust the amount of ethyl acetate according to the properties of raw materials, which aims to improving the yield and purity of the final product. In the research, using NIR spectra to reflect the comprehensive properties of coarse bear gall powder extracts, the process is optimized in a flexible way. Forty batches experiments are designed according to the weight ratio of ethyl acetate and coarse extracts of bear gall powder. The NIR spectra of the coarse extracts of bear gall powder are collected and processed using principal component analysis (PCA) method. The first 8 principal components combined with the amount of the ethyl acetate are used as the input variables, and calibration models are established to predict the yield and purity of the final product 30 batches are used as calibration set, which is used to establish the models, and other 10 batches are used as validation set, which is used for the performance appraisal of the established models. The correlation coefficients of the calibration, inner cross-validation and external validation for the purity model are 0.902, 0.896 and 0.883, respectively, and the RMSEC, RMSECV and RMSEP are 1.22%, 1.48% and 1.59%, respectively. The correlation coefficients of the calibration, inner cross-validation and external validation for the yield model are 0.921, 0.859 and 0.916, respectively, and the RMSEC, RMSECV and RMSEP are 1.39%, 1.65% and 1.53% respectively. This work demonstrated that NIR spectra combined with technology parameter could be used to predict the yield and purity of the final product. Using the established models, the most appropriate amount of the ethyl acetate can be determined according to the properties of the coarse bear gall powder extracts, and the yield and purity of the final product can be improved.
Acetates
;
chemistry
;
Animals
;
Gallbladder
;
chemistry
;
Medicine, Chinese Traditional
;
Powders
;
chemistry
;
Principal Component Analysis
;
methods
;
Spectroscopy, Near-Infrared
;
methods
;
Ursidae

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