Top Read Articles

    Published in last 1 year |  In last 2 years |  In last 3 years |  All
    Please wait a minute...
    For Selected: Toggle Thumbnails
    Chemical constituents of Murraya paniculata (L.) Jack.
    WANG Ru-ru, ZHANG Shan-shan, XU Hong-tao, WANG Yong-li, CHOU Gui-xin
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (5): 787-797.   DOI: 10.16333/j.1001-6880.2023.5.007
    Abstract758)      PDF(pc) (988KB)(20)       Save
    To study the chemical constituents of Murraya paniculata (L.) Jack, two new compounds, named as murrayanin (1) and 8-demethylnobiletin (2), along with twenty-four known compounds (3-26) were isolated from the 70% EtOH extract of M. paniculata by various chromatographic techniques such as silica gel, ODS, Sephadex-LH 20, Pre-HPLC, their structures was elucidated by spectral data analysis. The known compounds were identified as ficusal (3),lariciresinol-4′-monomethy ether (4),(±)-5′-methoxy-4′-O-methyllariciresinol (5),diospyrosin (6),(-)-9′-O-E-feruloyl-lyoniresinol (7),7-O-methylphellodenol-B (8),osthenon (9),3,4′-dihydroxy-3′,5′-dimethoxyphenylacetone (10),4′-hydroxy-5,7-dimethoxyflavanone (11),cystosiphonin (12),4′-hydroxy-5,7,3′-trimethoxyflavanone (13),5,7,3′,4′,5′-pentamethoxyflavanone (14),2′,4-dihydroxy-3′,4′,6′-trimethoxychalcone (15),2′,3-dihydroxy-4,4′,6′-trimethoxychalcone (16),evofolin B (17),2′-hydroxy-3,4,5,4′,6′-pentamoxychalcone (18),2′-hydroxy-3,4,4′,6′-tetramethoxychalone (19),5,8-dihydroxy-6,7,3′,4′-tetramethoxyflavone (20),3′-hydroxy-5,6,7,8,4′,5′-hexamethoxyflavone (21),5,3′,5′-trihydroxy-7,4′-dimethoxyflavone (22),5,7,3′-trihydroxy-8,4′-dimethoxyflavone(23),8-hydroxy-5,6,7,3′,4′-pentamoxyflavone (23),3′-hydroxy-5,6,7,4′-tetramethoxyflavone (24),5,7,3′,4′,5′-pentamethoxyflavone (25) and 5-hydroxy-6,7,8,3′,4′-pentamethoxyflavone (26),respectively.Compounds 3-8,10-13,15,16,20-24 were obtained from the genus Murraya for the first time,and compound 17 were isolated from M. paniculata for the first time.
    Related Articles | Metrics | Comments0
    Optimization of extraction process and composition analysis of essential oil from Amomum tsao-ko
    LIU Ju-zhao, XIAN Meng-xue, KONG Wei-hua, TIAN Xiao-li, YUAN Qiang, CUI Qi
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (5): 766-780.   DOI: 10.16333/j.1001-6880.2023.5.005
    Abstract533)      PDF(pc) (2439KB)(9)       Save
    In order to study the optimal extraction technology of essential oil from Amomum tsao-ko by steam distillation and ultrasonic organic solvent extraction.In this paper,based on single factor experiment,the extraction process of A. tsao-ko essential oil by steam distillation and ultrasonic organic solvent extraction was optimized by response surface methodology (RSM) and back-propagation artificial neural network (BP-ANN).Gas chromatography-mass spectrometry (GC-MS) was applied to analyze the essential oil components obtained from the two extraction methods,and the models were compared.The results showed that the optimum technological parameters of A. tsao-ko essential oil extraction by steam distillation were soaking time of 53.00 min,distillation time of 72.00 min,liquid to solid ratio of 11.00 mL/g,and extraction yield of 1.59%.A total of thirty-four components were identified in A. tsao-ko essential oil obtained by steam distillation.Under the optimum conditions of ultrasonic organic solvent extraction with ultrasonic power of 150.00 W,ultrasonic time of 27.00 min and liquid to solid ratio of 6.30 mL/g,the extraction yield was 3.40% and a total of 43 components were identified.Although the extraction yield of A. tsao-ko essential oil by ultrasonic organic solvent extraction was higher than that by steam distillation,the essential oil obtained by ultrasonic organic solvent extraction was poor in quality,dark in color and heavy in impurities,which was not conducive to the subsequent application.Compared with the RSM model,the BP-ANN model had a lower error-index and a higher fitting degree in predicting extraction yield,and had higher accuracy and prediction advantages.This study will provide reliable technological parameters for the efficient extraction of A. tsao-ko essential oil and further promote the development and utilization of the medicinal and edible plant A. tsao-ko.
    Related Articles | Metrics | Comments0
    Study on chemical constituents and anti-inflammatory activity of the pomace of Siraitia grosvenorii
    WU Juan-jiang, HUANG Hua-xue, LI Wen-chu, WANG Bin, GONG Li-min, SHENG Wen-bing, JIAN Yu-qing, WANG Wei
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1124-1134.   DOI: 10.16333/j.1001-6880.2023.7.003
    Abstract465)      PDF(pc) (843KB)(16)       Save
    The chemical constituents from the pomace fraction with water extract of Siraitia grosvenorii,and the anti-inflammatory activity in vitro were studied.The chemical constituents from the pomace fraction of Siraitia grosvenorii were isolated with various chromatographic separation techniques,and their structures were determined by entirely analyzing MS and NMR spectrums,there are oleanolic acid 28-O-β-D-glucopyranoside (1),oleanolic acid 3-O-β-D-glucuronyl-6′-ethyl ester (2),(3α)-7-oxomultiflor-8-ene-3,29-diol 3,29-dibenzoate (3),3,29-O-dibenzoyloxykarounidiol (4),isomultiflorenol (5),curcasinlignan B (6),urolignoside (7),diospyrosin (8),evofolin B (9),4′-hydroxyphenyl ethyl vanillate (10),aurantiamide (11),kaempferol-7-O-α-L-rhamnoside (12),kaempferitrin (13),kaempferol-3-O-β-D-glucopyranosyl-7-O-α-L-rhamnopyranoside (14),kaempferol 3-O-α-L-rhamnoside-7-O-β-D-xylosyl(1→2)-O-α-L-rhamnoside (15),sagittatin A (16),cucurbita-5,24-diene-3β-ol (17),balsaminol E (18),soya-cerebroside Ⅰ (19),β-sitosterol (20),β-daucosterol (21),7α-hydroxycholesterol (22),2E-4-hydroxy-nonenoic acid (23),vanillain (24),p-hydroxy-benzeneethanol (25),β-hydroxypropiovanillone (26).Compounds 2 and 6-11 were first isolated from Cucurbitaceae and compounds 1,3,4,14-16,18,19 and 22-26 were first isolated from Siraitia.Compounds 1-26 were preliminarily examined for anti-inflammatory activity in vitro,compounds 4,5,7,13,19 and 21 suppressed the production of TNF-α,compounds 13 and 21 both inhibited the production of IL-1β and IL-6,compounds 7,19 and 22 inhibited the production of IL-1β,compound 5 inhibited the production of IL-6.
    Related Articles | Metrics | Comments0
    Identification of UPLC characteristics spectrum of four original plants of Rabdosiae Herba
    XIAO Xiao-ji, QIU Jia-jia, LIU Jun-min, ZHAO Shuang-shuang, GU Jing-feng, WANG De-qin, ZHAN Ruo-ting
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 1038-1048.   DOI: 10.16333/j.1001-6880.2023.6.013
    Abstract429)      PDF(pc) (1619KB)(3)       Save
    The purpose of this paper is to establish the UPLC characteristics spectrum method for the identification of four original plants of Rabdosiae Herba,Rabdosia serra (Maxim.) H.Hara,Rabdosia lophanthoides (Buch.-Ham.ex D.Don) H.Hara,Rabdosia lophanthoides var. graciliflora (Benth.) H.Hara and Rabdosia stracheyi (Benth.ex Hook.f.) Hara.The similarity evaluation of characteristic chromatograms,cluster analysis (CA),principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA) were combined,which was used to study the common components and differential components of four original plants of Rabdosiae Herba.19,24,26 and 21 characteristic peaks were determined from the characteristic chromatograms of R. serraR.  lophanthoidesR. lophanthoides var. graciliflora and R. stracheyi.In addition to R.serra,the similarity of the samples of the other three original plants was high,but there were some differences in similarity between different original plants.R. serra could be clearly distinguished from the other three original plants by CA and PCA and OPLS-DA could be used to distinguish R. lophanthoides, R. lophanthoides var. graciliflora and R. stracheyi.The UPLC characteristics spectrum established in this study combined with chemometrics comprehensively reflected the chemical composition of four original plants of Rabdosiae Herba,and the method is simple,rapid and specific,which can provide a reference for the identification,quality analysis and evaluation of original plants of Rabdosiae Herba.
    Related Articles | Metrics | Comments0
    Quality difference between Fritillariae Thunbergii Bulbus and Fritillariae Hupehensis Bulbus based on HPLC-ELSD fingerprints and multi-component quantitative analysis
    LIANG Yue-yi, LI Zhen-yu, LYU Wei-sheng, LU Xiao-ying, YANG Jie, LIU Xiao-xia, WEI Cui-jie, SUN Dong-mei
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1101-1111.   DOI: 10.16333/j.1001-6880.2023.7.001
    Abstract373)      PDF(pc) (1746KB)(4)       Save
    The HPLC-ELSD fingerprints of Fritillariae Thunbergii Bulbus (FTB) and Fritillariae Hupehensis Bulbus (FHB) were established to compare the differences between the two herbs combined with multi-component quantitative analysis.The chromatographic column was Waters ACQUITY HSS T3 (4.6 mm×250 mm,5 μm).The gradient elution was performed at a flow rate of 1.1 mL/min with acetonitrile -0.1% triethylamine solution as the mobile phase.The column temperature was 38 ℃.Evaporative light scattering detection was used.The HPLC-ELSD fingerprints of FTB and FHB were established.The differences between FTB and FHB were compared by chemometrics methods and simultaneous determination of five alkaloid components.The results showed that the fingerprints of FTB had seven common peaks and the fingerprints of FHB had eight peaks.Six peaks were identified,namely yibeissine,peimisine,peimine,peiminine,isofritillarine and hupehenine.Among the above ingredients,hupehenine was the specific component of FHB.Both HCA and PCA could distinguish FTB and FHB well.OPLS-DA found four different markers.The content determination results showed that the content of peimine in FTB was significantly higher than that in FHB,while the contents of peimisine,peiminine and isofritillarine were significantly lower than that in FHB.This method can effectively identify the quality differences between FTB and FHB,and provide reference for their quality control.
    Related Articles | Metrics | Comments0
    Research progress on antioxidative property of ergothioneine and its intervention in oxidative stress-related diseases
    GAO Qing-ying, XU Jian-xiong
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 1081-1087.   DOI: 10.16333/j.1001-6880.2023.6.017
    Abstract351)      PDF(pc) (670KB)(23)       Save
    Ergothioneine (ERG) is a kind of sulfur-containing amino acid existing in nature.It has been widely used in food,medicine,cosmetics and other fields in recent years for the reasons that it has strong ability of scavenging free radicals,anti-aging and the characters of natural,safe,non-toxic.As a natural and efficient antioxidant,ergothioneine plays an important role in the prevention and treatment of diseases related to oxidative stress.This paper reviewed the structure,properties and antioxidant properties,as well as progress in search of the role and mechanism of ergothioneine in the intervention of diseases related to oxidative stress,which provides a reference for the multiple application of ergothioneine.
    Related Articles | Metrics | Comments0
    Mechanism of Phellodendri Chinensis Cortex in the treatment of gout based on network pharmacology,molecular docking and experimental validation
    LI Min, LI Li, QUAN Yun-yun, ZENG Jin, ZHAO Jun-ning, MAO Jiu-zhou, GONG Xiao-li, YIN Zhu-jun
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1235-1246.   DOI: 10.16333/j.1001-6880.2023.7.015
    Abstract315)      PDF(pc) (2380KB)(32)       Save
    To investigate the potential pharmacodynamic material basis and mechanism of Phellodendri Chinensis Cortex (PCC) in the treatment of gout by combining network pharmacology prediction,molecular docking validation and experimental validation.The active ingredients and action targets of PCC were obtained through TCMSP databases,gout-related disease targets were obtained from GeneCards,OMIM and TTD databases.The corresponding targets of the active ingredients of PCC were intersected with the gout targets,and the protein-protein interaction (PPI) network of the intersected genes was mapped with the help of STRING platform and Cytoscape 3.9.0 software.The analysis of gene ontology (GO) function and Kyoto gene and gene targets (KEGG) pathway enrichment were performed by using String and MetaScape databases and visually presented through the platform of bioinformatics.Furthermore,molecular docking technology was performed to validate the binding pattern and affinity between the key ingredients and the crucial targets by using AutoDock Tools software.A total of 25 active ingredients and 70 potential key targets for the treatment of gout was screened in PCC. The enrichment of GO function and KEGG pathway showed that PCC might positively regulate cell migration,negatively regulate cell differentiation,inflammatory response,positively regulate cell adhesion,protein phosphorylation,DNA transcription and other biological processes.The most crucial biotargets of PCC against gout were protein kinase B1 (AKT1),tumor necrosis factor (TNF),peroxisome proliferative activated receptor gamma (PPAR γ),interleukin-6 (IL-6),prostaglandin-endoperoxide synthase 2(PTGS2),and KEGG enrichment analysis revealed that PCC possessed anti-gout activity by regulating PI3K-Akt signaling pathway,MAPK signaling pathway.The molecular docking results showed that the binding energy between the key biotargets and the five potential active components were much less than -5 kcal/mol.In vitro experiments showed that the core chemical components exhibited potent inhibitory effect on the inflammatory response induced by sodium urate crystal.This study initially revealed PCC has a variety of potential anti-gout active components,and its mechanism may be achieved by modulating multiple biotargets and multiple signal transduction pathways.
    Related Articles | Metrics | Comments0
    Quality analysis of Hirudo with different origins and its adulterants
    LI Guo-wei, QIU Yun-jing, TONG Pei-zhen, HU Qi-ping, DENG Li-ping, YANG Li, SUN Dong-mei
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 997-1006.   DOI: 10.16333/j.1001-6880.2023.6.009
    Abstract314)      PDF(pc) (1321KB)(7)       Save
    HPLC method was used to establish the characteristic chromatogram of Hirudo and the contents of 5 components were determined,which provides a basis for quality evaluation of different origins Hirudo and its adulterants.The Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version) was used to analyze the results of the characteristic chromatogram.The results showed that there were eight common peaks in 27 batches of Whitmania pigra Whitman and five peaks were identified by comparison with reference substances,respectively,which were uracil,hypoxanthine,xanthine,hirudine C carboxyl derivatives and hirudine B.The SIMCA-P 14.0 software were used for cluster analysis and partial least squares discriminant analysis(PLS-DA) that both of them can distinguish Hirudo of different origins and its adulterants into four different classes.In addition,the content determination results showed that there were some differences in the content of Hirudo with different origins and its adulterants.The average content of uracil and hypoxanthine was in the order of Whitmania pigra Whitman>Hirudo nipponica Whitman>Whitmania acranulata Whitman>Poecilobdella manillensis Lesson;the average content of xanthine was in the order of Whitmania pigra Whitman>Hirudo nipponica Whitman>Poecilobdella manillensis Lesson>Whitmania acranulata Whitman;the average content of hirudine C carboxyl derivatives was in the order of  Whitmania acranulata Whitman>Whitmania pigra Whitman>Hirudo nipponica Whitman>Poecilobdella manillensis Lesson;the average content of hirudine B was in order of Whitmania acranulata Whitman>Whitmania pigra Whitman>Hirudo nipponica Whitman>Poecilobdella manillensis Lesson.The method established in this study can be used as a quality evaluation method for Hirudo with different origins and its adulterants,and provide a reference for the quality evaluation of Whitmania pigra Whitman.

    Related Articles | Metrics | Comments0
    Mechanism of olibanum in the treatment of acute kidney injury based on network pharmacology and in vitro cell experiment verification
    BAI Juan, ZHANG Yan, HONG Shu-xia, SHI Jin-ping, XU Li-ting, WANG Fang
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (9): 1613-1623.   DOI: 10.16333/j.1001-6880.2023.9.015
    Abstract307)      PDF(pc) (2101KB)(110)       Save
    To study the mechanism of olibanum on acute kidney injury (AKI) based on network pharmacology,and establish an in vitro model of human renal tubular epithelial cells (HK-2) according to the results.To search 223 drug targets through the TCMSP database and the Swiss Target Prediction database,then using the OMIM,DisGeNet,DrugBank and GeneCards database to collect 1 245 disease targets,72 drug-disease targets were obtained through the Venn diagram tool.Protein interaction PPI network was constructed by String database;GO analysis and KEGG pathway analysis were performed using DAVID database;Cytoscape 3.7.2 software was used to construct the network diagram;Molecular docking was performed by Autodock.Involving Toll-like receptor、TNF、NF-kappa B and other signaling pathways.Based on this,the proliferation of HK-2 was detected by cell counting kit-8 (CCK-8) method,and the grouping was determined.The rate of cell apoptosis was detected by Hoechst33258 fluorescent staining,and the quantitative real time polymerase chain reaction (Q-PCR) was used to detect the relative expression of MAPK3 and PPARG mRNA.The experimental results showed that in CCK-8 experiment,compared with the model group,the cell viability in low,medium and high dose groups decreased gradually (P <0.05);Hoechst33258 experiment,ELISA experiment and Q-PCR experiment,compared with the model group,the cell apoptosis rate,the capacity of cellular inflammatory factors and the molecular weight and mRNA expression of MAPK3 and PPARG protein decreased significantly (P <0.05).In conclusion,olibanum has the characteristics of multi-target and multi-pathway,which provides a basis for the clinical application of this drug.
    Related Articles | Metrics | Comments0
    Total components analysis of Zanthoxylum bugeanum seeds from different orgins
    WU Juan-juan, ZHANG Jia-yu, SHI Shi-lei, LIAN Yun-he, CHENG Yuan-xin, WU Ya-ming, CAI Xing-fu, WU Di
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (5): 837-851.   DOI: 10.16333/j.1001-6880.2023.5.012
    Abstract299)      PDF(pc) (2753KB)(13)       Save
    Zanthoxylum bungeanum seeds are the main byproduct of Z. bungeanum.At present,the researches of Z. bungeanum seeds mainly focus on lipid,and less researches on other chemical components.The chemical composition and nutrient composition of Z. bungeanum seeds from Jinyang,Hanyuan and Wudu were analyzed using chemical method and chromatographic technique to fully exploit nature resources.Results indicated that 92 volatile compounds and 28 non-volatile compounds were identified in three samples.The volatile components were mainly alcohols,carbonyls,esters,terpenes,of which only seven compounds were same components,and the non-volatile components were mainly phenols,alkaloids,fatty acids,coumarins,lignans,of which 18 compounds were same components.Meanwhile,the nutritional composition,including protein,oil and sugar,were analyzed in this paper.Through the systematic research of this paper,the material basis and research ideas were provided to further develop the edible and commercial value of Z. bungeanum seeds.
    Related Articles | Metrics | Comments0
    Research progress on pharmacological action and mechanism of Ganoderma lucidum polysaccharides
    ZHANG Ruo-bing, YANG Yu-he, LI Chen-xue, LI Wen-lan
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (5): 879-887.   DOI: 10.16333/j.1001-6880.2023.5.016
    Abstract299)      PDF(pc) (1281KB)(18)       Save
    Ganoderma lucidum is an important dual-use fungus with a long history both at medicinal and edible.At present,about 100 species of G. lucidum are known.Polysaccharide is one of the main active ingredient of G. lucidum.It can inhibit the proliferation and differentiation of tumor cells,regulate congenital immune cells,microbial groups and the expression of inflammatory factors in the body,and terminate reactions of free radical chain.It can also increase the vitality of cells and antioxidant enzymes,regulate the metabolism of lipids and organic compounds in the liver,inhibit the apoptosis of myocardial cells,inhibit the activation of microglia and astrocyte,etc.Thus,it has pharmacological effects such as anti-tumor,immunomodulatory,anti-inflammatory, hypoglycemic,hypolipidemic, anti-fatigue,anti-oxidation,protection myocardial and liver,regulating central nervous system and so on.In this paper,the pharmacological effects and mechanisms of G. lucidum polysaccharides in recent three years are reviewed,with a view to provide reference for further development and utilization of G. lucidum polysaccharides.
    Related Articles | Metrics | Comments0
    Study on HPLC fingerprint and chemical pattern recognition of triterpenoids in Poriae Cutis and White Poria
    ZHAN Hui-hui, DING Chan, PENG Si-yuan, LIU Yuan, MENG Jun-hua, XIAO Zuo-wei, CUI Pei-wu
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (9): 1469-1479.   DOI: 10.16333/j.1001-6880.2023.9.001
    Abstract276)      PDF(pc) (1323KB)(50)       Save
    In this study,HPLC analysis combined with chemical pattern recognition was employed to establish the HPLC fingerprint related to triterpenoids in Poriae Cutis and White Poria,which can provide a reference for quality control and standard establishment of decoction pieces derived from mycomedicine Poria cocos.HPLC method was adopted and performed on an Agilent 5 TC-C18 (2) column (250 mm × 4.6 mm,5 μm).The mobile phase was consisted of acetonitrile and 0.3% phosphoric acid solution,and gradient elution procedure was employed with the flow rate setting at 1.0 mL/min.The column temperature was set at 25 °C,the detection wavelength was set at 242 and 203 nm,and the injection volume was 10 μL,respectively.To establish the HPLC fingerprint and chemical pattern recognition of Poriae Cutis and White Poria,cluster analysis (CA),principal component analysis (PCA) and similarity evaluation were adopted to process the experimental data,the similarity and difference of Poriae Cutis and White Poria samples were analyzed subsequently.Finally,the key chromatographic peaks representing triterpenoids were pointed out from the established HPLC fingerprint.Among the common chemical components showed in the chromatograms,six peaks related to poricoic acid B (peak 2),dehydrotumulosic acid (peak 3),poricoic acid A (peak 4),dehydropachymic acid (peak 9),pachymic acid (peak 10),and dehydrotrametenolic acid (peak 10) were indentified,respectively.The similarity of tested decoction pieces from the same part of P. cocos sclerotium were all above 0.90.According to CA and PCA data,all the decoction pieces analyzed in this study can be divided into two groups representing different medicinal parts of P. cocos sclerotium.Seven common chromatographic peaks including peak 3 (dehydrotumulosic acid),peak 5,peak 6,peak 8,peak 9 (dehydropachymic acid),peak 10 (pachymic acid) and peak 11 were essential for the quality evaluation of Poriae Cutis and White Poria,the other eight chromatographic peaks including peak 1,peak 2 (poricoic acid B),peak 4 (poricoic acid A),peak 7,peak 12 (dehydrotrametenolic acid) and peaks 13 to 15 could be used as the characteristic identification peaks for evaluating the quality of Poriae Cutis according to the loading scatter analysis of PCA data.The characteristic difference deduced from the 15 common chromatographic peaks of White Poria samples was smaller than that of Poriae Cutis samples.The established HPLC fingerprint of Poriae Cutis and White Poria combined with chemical pattern recognition can provide a scientific reference for the quality control and evaluation of samples or products derived from P. cocos .

    Related Articles | Metrics | Comments0
    Research progress on chemical constituents,pharmacological activities and quality control methods of Clerodendranthus spicatus
    ZHAO Yu-hui, XIANG Cheng, XU Tian-rui, HAO Qian
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1247-1264.   DOI: 10.16333/j.1001-6880.2023.7.016
    Abstract268)      PDF(pc) (2224KB)(1)       Save
    As a well-known national medicine,Clerodendranthus spicatus has a long history medicinal in our country.It is rich in flavonoids,phenylpropanoids and terpenoids and other chemical components,with heat-clearing and dampness-removing,stone-removing diuresis,anti-inflammatory,antioxidant and antibacterial and other pharmacological activities,the authenticity of kidney tea is very important to the efficacy.This paper reviews the chemical composition,pharmacological activity and quality control methods of C. spicatus,in order to provide reference for the further study of the chemical composition and pharmacological activity of kidney tea and the detection of the quality of C. spicatus.
    Related Articles | Metrics | Comments0
    Optimization of fermentation process of paraherquamide A by response surface methodology
    CHEN Xiao-na, PENG Jia-fei, WANG Pin-mei, XU Jin-zhong
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 1049-1057.   DOI: 10.16333/j.1001-6880.2023.6.014
    Abstract252)      PDF(pc) (1168KB)(21)       Save
    Paraherquamide A(PA) is an important raw material for the new anthelmintic drug Startect®,and its low yield limits the promotion and use of the drug.Response surface methodology (RSM) was used to optimize the fermentation process of wild Penicillium sp.KWF31 to produce PA.Eight factors of the medium were screened by Plackett-Burman test design.Among them,the dosages of soluble starch,glucose and yeast extract were found to be the key factors.The optimal concentration ranges of the three factors were examined by the steepest ascent path and the Box-Behnken design as 31.0 g/L soluble starch,12.8 g/L glucose and 14.5 g/L yeast extract,respectively.Under the optimal condition,the maximum titer of PA approached 243.7 mg/L,which was close to the theoretical value of 241.6 mg/L,and 3.4 times of the initial.
    Related Articles | Metrics | Comments0
    Study on the effect of asperosaponin VI on neurogenesis and cognitive function in sleep-deprived mice
    WANG Mei, LIU Yu-e, SU Da-peng, WANG Ying-xue, CHEN Hong-mei, DONG Xing-xing, HUANG Shen-rong, ZHANG Jin-qiang
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (5): 733-740.   DOI: 10.16333/j.1001-6880.2023.5.001
    Abstract249)      PDF(pc) (1218KB)(3)       Save
    The aim of this study was to investigate the effect of asperosaponin VI (ASA VI) on cognitive function in sleep-deprived mice and the related mechanisms.C57BL/6J mice were subjected to sleep deprivation by multi-platform water environment method,during which the mice were intraperitoneally injected with ASA VI.New object recognition test and Morris water maze test were used to evaluate the cognitive function of mice.The changes in the inflammatory levels,neurogenesis and signaling pathways in hippocampus were detected by q-PCR,immunohistochemistry and Western blotting.The results showed that compared with the control group,microglia in the hippocampus of the sleep-deprived mice were activated,the expression level of inflammatory cytokines was significantly increased (P < 0.05),the number of newborn neurons was significantly decreased (P < 0.05),and the cognitive function was decreased.The ASA VI intervention group significantly improved the cognitive ability of sleep-deprived mice,inhibited the expressions of pro-inflammatory cytokines IL-1β,TNF-α and IL-6 in hippocampus (P < 0.05),and significantly increased the expression levels of anti-inflammatory cytokines IL-4 and IL-10 and neurotrophic factor BDNF (P < 0.05).At the same time,the protein expression levels of p-PI3K,PI3K and Akt,as well as the number of new neurons in hippocampus were increased by the intervention of ASA VI (P < 0.05).Treatment with PI3K/Akt inhibitor LY294002 significantly reduced the intervention effect of ASA VI on sleep-deprived mice.These results suggested that ASA VI could improve the learning and memory of sleep-deprived mice,and its mechanism was related to the activation of PI3K/Akt signaling pathway.ASA VI has potential development prospects as an anti-inflammatory and neuroprotective drug.
    Related Articles | Metrics | Comments0
    Isolation and purification of Qi-Gui polysaccharides and their in vitro inhibitory activity on tumor cell proliferation
    LIU Wen-juan, MA Shan-bo, LU Wen-fei, CUI Dong-xiao, XU Ding-qiao, YUE Shi-jun, CAO Wei, TANG Yu-ping
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (10): 1651-1658.   DOI: 10.16333/j.1001-6880.2023.10.001
    Abstract249)      PDF(pc) (1607KB)(35)       Save
    Astragali Radix-Angelica Sinensis Radix (Qi-Gui herb pair) has long been used as a classic herb pair for tonifying qi and nourishing blood.Polysaccharide is one of its major active ingredients.Two of the Qi-Gui polysaccharides named AAPS-1a and AAPS-3a,significantly inhibited the proliferation of human colon cancer cells HCT116 and human lung carcinoma cells A549,with IC50 of (47.59±1.3) μg/mL,(18.33±2.5) μg/mL for HCT116 cells,and IC50 of (52.12±1.1) μg/mL,(170.21±2.3) μg/mL for A549 cells.Furthermore,AAPS-1a and AAPS-3a induced apoptosis in HCT116 and A549 cells.The apoptotic rate of the HCT116 cells were 31.4%,46.5% respectively and the apoptotic rate of the A549 cells were 19.3% and 33.0% separately.Structural analysis showed that AAPS-1a and AAPS-3a were homogeneous polysaccharides with molecular weights of 7.3×105 and 8.5×104 Da,respectively.Our study found two homogeneous Qi-Gui polysaccharides with significant anti-tumor activity in vitro,which provide experimental basis for clarifying the anti-tumor structure of polysaccharides in Qi-Gui herb pair.
    Related Articles | Metrics | Comments0
    Secondary metabolites of endophyte Paraconiothyrium sp.YLHJ01 from Artemisia selengensis
    ZENG Qing-ting, YUAN Tao
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (5): 781-786.   DOI: 10.16333/j.1001-6880.2023.5.006
    Abstract248)      PDF(pc) (601KB)(21)       Save
    Seven compounds were isolated from the extract of the endophytic fungus Paraconiothyrium sp.YLHJ01,their structures were identified as one new compound,2-(1S,3R-dihydroxybutyl)benzene-1,4-diol (1) and six known compounds, 6-hydroxy-2S-methyl-4-chromanon (2),10-norparvulenone (3),(S)-7-hydroxy-3-((S)-1-hydroxyethyl)isobenzofuran-1(3H)-one (4),murranoic acid A (5),modiolide G (6),and uridine (7) by the analysis of spectroscopic data including NMR,ECD,IR and HR-ESI-MS.The cytotoxic activity in vitro assay indicated that none of the compounds showed cytotoxic activity on A549 and HepG2 cells. Antibacterial assay revealed that compounds 1 and 4 showed weak inhibitory activities against Staphylococcus aureus with minimum inhibitory concentrations (MICs) of 400 and 100 μg/mL,respectively,while compound 2 showed weak inhibition against S. aureus and Escherichia coli with MICs of 800 and 400 μg/mL,respectively.
    Related Articles | Metrics | Comments0
    Quality analysis of Nymphaeae Flos based on HPLC fingerprint and QAMS
    ZUO Di-jun, LI Chen-yang, YANG Jun, XU fang, ZHAO Jun
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1112-1123.   DOI: 10.16333/j.1001-6880.2023.7.002
    Abstract248)      PDF(pc) (1626KB)(3)       Save
    To establishthe HPLC fingerprint of Nymphaeae Flos and Quantitative analysis of multi-components by single-marker (QAMS) for seven contents.The HPLC fingerprint was established by Phenomenex Gemini NX-C18 (250 mm×4.6 mm,5 μm).The mobile phase was acetonitrile-0.2% phosphoric acid aqueous solution.The flow rate was 1.0 mL/min.The column temperature was 30 ℃.The detection wavelength were 266 nm. The data were studied by Similarity analysis,cluster analysis and principal component analysis.Gallic acid was used as an internal standard to calculate the relative correction factors of the other six constituents.A total of eleven common peaks were identified and seven components were identified for the analysis of 17 batches of Nymphaeae Flos.The similarity of 17 batches of samples was greater than 0.9.The HPLC similarity of two commercial samples and S16 was 0.568 and 0.730,which the results show that commercial Nymphaeae Flos were different from cultivated and wild Nymphaeae Flos.Principal component analysis,cluster analysis and orthogonal partial least squares discriminant analysis (OPLS-DA) were confirmed this conclusion.With gallic acid was the reference,the relative correction factors of methyl gallate,isostrictiniin,geraniin,ellagic acid,nicotiflorin,pentagalloylglucose were 0.965 0,0.974 0,1.201 3,0.175 4,1.160 3 and 2.117 3,RSD were 0.37%,0.38%,0.32%,1.76%,0.37% and 0.08% (n = 8).At the same time,the result obtained by QAMS approximated those obtained by external standard method (ESM).The established fingerprint and QAMS content determination method of Nymphaeae Flos were feasible and provide a reference for the improvement of the quality control method of Nymphaeae Flos.
    Related Articles | Metrics | Comments0
    Chemical composition analysis of mango kernels by UPLC-Q-TOF-MS/MS
    WU Jun-ying, ZHANG Wu-gang, LANG Yi-fan, HUANG Zhi-chao, YANG Wu-liang, YANG Jie , CHEN Hai-fang
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 949-965.   DOI: 10.16333/j.1001-6880.2023.6.004
    Abstract247)      PDF(pc) (1564KB)(10)       Save
    To systematically investigate the chemical constituents of mango kernels,ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was analyzed in this study.Electrospray ion source (ESI) was adopted to collect the data in negative ion mode.A total 135 compounds were identified according to analyze the mass data from the references and related literatures,including 47 berberine-type alkaloids,47 flavonoids,30 organic acids and three coumarins,eight iridoid glycosides.Seventeen compounds were identified in this genus for the first time.Using UPLC-Q-TOF-MS/MS technology to analyze the chemical constituents of mango kernels,which will provide a significant basis for the subsequent study of material basis and clinical application.
    Related Articles | Metrics | Comments0
    Chemical constituents of Senecio scandens and their anti-tobacco mosaic virus activity
    FU Tao, LI Ya-li, ZHANG Wei, WU Jiang-mei, LIU Yang, LUO Xin-xiang, YAN Xiao-hui, HU Shi-jun
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (9): 1554-1561.   DOI: 10.16333/j.1001-6880.2023.9.009
    Abstract239)      PDF(pc) (778KB)(42)       Save
    In order to provide a theoretical basis for the development of new plant virus inhibitors,the chemical components of Senecio scandens and its activity against tobacco mosaic virus (TMV) were determined.The chemical constituents of the ethyl acetate extract from the aboveground part of S. scandens were separated by silica gel,Sephadex LH-20,MCI and other column chromatography methods,and its structures were identified by NMR and MS data.The inhibitory activity of the compound against TMV was screened by the half-leaf spot method in vivo.Thirteen compounds were isolated from the ethyl acetate extract of S. scandens. According to their physicochemical properties and spectral data,they were identified as 9,19-cyclolanost-24-en-3-one (1),7β-methoxystigmast-5-en-3β-ol (2),dotriacontane (3),ursolic acid (4),stigmasterol (5),(22E)-ergosta-6,22-diene-3β,5β,8α-triol (6),kaempferol (7),indole-3-carboxaldehyde (8),nepetin (9),quetrcetin (10),taxifolin (11),jacaranone (12),phytol (13).Except compound 7 and compound 10,the other compounds were isolated from this plant for the first time.The results showed that compound 8 and compound 13 had strong anti-TMV activity.
    Related Articles | Metrics | Comments0
    Mechanism of active ingredients from Scutellaria Radix on alcoholic liver disease based on network pharmacology and molecular docking and effect verification
    YE Jing-rong, LIU Rui, CHENG Cheng, ZHANG Feng-ying, YANG Xue
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (9): 1602-1612.   DOI: 10.16333/j.1001-6880.2023.9.014
    Abstract237)      PDF(pc) (2851KB)(60)       Save
    Network pharmacology and molecular docking technology were adopted to explore the possible mechanism of Scutellaria Radix in the treatment of alcoholic liver disease.The therapeutic effect of active ingredient of Scutellaria Radix on alcoholic liver disease was verified by cell experiment in vitro.The effective components and targets of Scutellaria Radix were retrieved from TCMSP,Swiss ADME and Swiss Target Prediction databases;The disease targets related to alcoholic liver disease were retrieved from GeneCards,OMIM,DisGeNET,TTD and PharmGKB databases;Using String database to build target interaction network;Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and gene ontology (GO) enrichment analysis were conducted for key targets through metascape database.The "active ingredient target pathway" interaction network of Scutellaria Radix for the treatment of alcoholic liver disease was constructed by using Cytoscape 3.8.0 software,and the active ingredient and key target of Scutellaria Radix were screened for molecular docking.Based on the results of network pharmacology and molecular docking,the prediction results were preliminarily verified by cell experiments in vitro.A total of 27 active components of Scutellaria Radix were obtained after ADME screening,and these 27 active components could play a therapeutic role in alcoholic liver disease through 257 gene targets,among which the key core targets are SRC,AKT1,PIK3R1,STAT3,PIK3CA,etc.The enrichment analysis of KEGG signal pathway showed that the main signal pathways of Scutellaria Radix in treating alcoholic liver disease included cancer pathway,PI3K Akt signal pathway,lipid and atherosclerosis,chemical carcinogenic reactive oxygen species,prostate cancer,etc;Molecular docking results suggest that Alpinetin may be one of the key effective ingredients of Scutellaria Radix in the treatment of alcoholic liver disease;In vitro cell experiments proved that Alpinetin could significantly improve the alcoholic injury of rat liver cells BRL3A.Through the results of network pharmacology,molecular docking technology and cell experiment,it can be analyzed and inferred that Scutellaria Radix can play a role in the prevention and treatment of alcoholic liver disease through multi-component and multi target methods.As a key active ingredient,Alpinetin can provide basis and reference for further development of drugs related to the treatment of alcoholic liver injury by using Scutellaria Radix.
    Related Articles | Metrics | Comments0
    Chemical constituents from Mongolian medicine Inulae Radix
    MAO Ting, MAO Le, ZHANG Ping, GAO Bai-qing, ZHANG Yong
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 973-980.   DOI: 10.16333/j.1001-6880.2023.6.006
    Abstract231)      PDF(pc) (524KB)(14)       Save

    To study the chemical constituents from the roots of the Mongolian medicine Inulae Radix.The compounds were separated and purified by RP-HPLC and silver nitrate silica gel chromatography,and the structures of the compounds were identified by applying NMR and MS spectroscopy.Twenty-two monomeric compounds were isolated and identified as follows:5α,6α-dihydroxy-eudesma-11(13)-dien-12,8β-olide(1),4α-hydroxy-5-methoxy-5,8α-dimethyl-3-methylenedecahydronaphtho[2,3-b]furan-2-one (2),1(10),4(5),11(13)-germa-cradiene-8,12-olide (3),alantolactone (4),isoalantolactone (5),11,13-dihydro-5(6)-dine-alantolactone (6),alloalantolactone (7),4α,5β-epoxy-1(10),11(13)-germacra-dien-12,8α-olide (8),sseptuplinolide (9),8-epi-ivangustin (10),santamarine (11),racemosalactone A (12),3-oxo-eudesma-4(11)-dien-12,8β-olide (13),telekin (14),igalane (15),macrophyllilactone E (16),4α,15-epoxy-isoalantolactone (17),11,13-dihydroxy-alantolactone (18),1(2),4(15),11(3)-eudesma-trien-12,8β-olide (19),linolenic (20),linoleic acid (21),oleic acid (22).Compound 1 was a new sesquiterpenoes.Compounds 2,3,9-14,17,19-22 were isolated from genus Inula for the first time.Compounds 4-8,15,16,18 have been reported from Inulae Radix.

    Related Articles | Metrics | Comments0
    Research progress on compositions and pharmacological activities of essential oil from Chimonanthus Lindl.
    HE De-ying, XIAO Wei-yi, LI Cheng, ZOU Zheng-rong
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (9): 1624-1636.   DOI: 10.16333/j.1001-6880.2023.9.016
    Abstract230)      PDF(pc) (1502KB)(58)       Save
    Chimonanthus Lindl.is a unique traditional medicinal plant in China,with a wide distribution and abundant resources.The essential oil is one of the main active components of the plant,including terpenoids,aromatic compounds and aliphatic compounds,which have pharmacological activities such as antibacterial,anti-inflammatory,antioxidant,antiviral and cough suppressant.It has good therapeutic effects on diseases such as vascular dementia,acute lung injury and ulcerative colitis.This paper presents a systematic classification of the chemical structures and synthesis pathways of essential oil components of Chimonanthus Lindl.and provides a brief overview of the related biological activities in order to make references for the further development and utilization of Chimonanthus Lindl.
    Related Articles | Metrics | Comments0
    Optimization of the extraction process of total phenolic acid from Trollii Flos by response surface methodlogy and study on its whitening activity
    DU Ruo-jun, SHI Zhao-hua, ZHAN Zhi-lai, HU Jun-jie, ZHENG Guo-hua
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 915-924.   DOI: 10.16333/j.1001-6880.2023.6.001
    Abstract228)      PDF(pc) (2087KB)(22)       Save
    Box-Behnken design experiments in response surface analysis were used to optimize the process conditions for the ethanol reflux extraction of total phenolic acid from Trollii Flos,and the scavenging ability of total phenolic acid from Trollii Flos on DPPH radicals and hydroxyl radicals and its inhibitory effect on tyrosinase were also studied.The results showed that the optimum extraction process of total phenolic acid from Trollii Flos was:ethanol concentration 60%,reflux time 90 min,reflux temperature 60 ℃,liquid to solid ratio 40∶1 (mL/g).The yield of total phenolic acid from Trollii Flos under these conditions was 92.245 mg/g.In addition,it had certain scavenging ability to DPPH radicals and hydroxyl radicals,and could significantly inhibit tyrosinase activity.The extract method is stable and reliable,and the total phenolic acid from Trollii Flos has certain in vitro antioxidant activity and tyrosinase inhibitory effect,which lays the foundation for further scientific and rational development and utilization of Trollii Flos.
    Related Articles | Metrics | Comments0
    Protective effects and mechanism of Moringa oleifera leaf polysaccharide on ulcerative colitis in mice
    Hosameldeen Mohamed Husien, PENG Wei-long, LIU Ming-jiang, BO Ruo-nan, LI Jin-gui
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (10): 1659-1668.   DOI: 10.16333/j.1001-6880.2023.10.002
    Abstract226)      PDF(pc) (1469KB)(20)       Save
    The aim of this study was to investigate the preventive and therapeutic effects of Moringa oleifera leaf polysaccharide (MOLP) on ulcerative colitis (UC) in mice induced by dextran sodium sulfate (DSS).50 BALB/c mice were randomly divided into normal Con group,DSS group,MOLP-L(25 mg/kg) group,MOLF-M (50 mg/kg) group,and MOLF-H(100 mg/kg) group.Mice were induced UC by drinking 4% DSS.During the experiment,the general condition and body weight of mice were recorded daily,and the changes in disease activity index (DAI) were observed;ELISA was used to measure the contents of TNF-α,IL-1β,IL-10,HMGB1 in serum and MPO in colon tissue;HE staining was used to observe the pathological morphological damage of colon tissues;qPCR was used to detect TNF-α,IL-1β,IL-10 and HMGB1 mRNA expression levels in colon tissues;Western blot was used to measure TLR4,MyD88,P65,p-P65,IκBα and p-IκBα expression in colon tissues;16S rRNA was showed the changes of intestinal microflora in mice.Compared with the Con group,mice in the DSS group had decreased general condition,weight loss,colon length was shortened,DAI and pathological section damage scores were significantly increased (P<0.001),TNF-α,IL-1β,HMGB1 content and MPO activity were significantly increased (P<0.001),TLR4,MyD88,colon tissue.The protein expression levels of p-P65 and p-IκBα were significantly upregulated (P<0.001);Compared with the DSS group,the condition of mice in the different doses of MOLP treatment group was significantly improved,and significantly increased IL-10 expression,the results were dose-dependent.MOLP also could regulate the gut microbiota diversity significantly and restore the balance of intestinal bacteria.The results indicated that MOLP might inhibit inflammatory response and regulate the diversity,composition and relative abundance of gut microbiota in mice,which could play a positive role in preventing and treating UC.
    Related Articles | Metrics | Comments0
    Effect of decocting time on chemical composition and anti-fatigue effect of Triphala decoction
    DENG Xuan, QIU Min, HUANG Hao-zhou, FAN San-hu, ZHANG Ding-kun, LUO Chuan-hong, HAN Li, LIN Jun-zhi
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1212-1221.   DOI: 10.16333/j.1001-6880.2023.7.013
    Abstract222)      PDF(pc) (2476KB)(8)       Save
    To explore the effect of decocting time on the component transformation and anti-fatigue effect of Triphala decoction.HPLC method was used to track the changes of ingredients in Triphala decoction during decoction.Twelve differential compounds were screened through partial least squares discriminant analysis (PLS-DA) and seven of these were identified as gallic acid,chebulic acid,chebulinic acid,chebulagic acid,ethyl gallate,ellagic acid and corilagin.During the decoction,the contents of chebulinic acid,chebulagic acid,ethyl gallate and ellagic acid all decreased while the gallic acid and chebulic acid increased.Corilagin′s content increased first,then reduced.The anti-fatigue effect of Triphala decoction with different extraction times was then evaluated.The longer the decoction time,the more potent the anti-fatigue effect was,according to the results of swimming time and 11 fatigue-related markers in mice.Gallic acid and chebulic acid,among the differential compounds,have the greatest impact on the anti-fatigue effect.It demonstrates how decocting can encourage the hydrolysis of the macromolecular tannins in Triphala decoction into gallic acid and chebulic acid,which can improve the anti-fatigue effect of Triphala decoction.
    Related Articles | Metrics | Comments0
    Mechanism of Erzhi Pill against Alzheimer′s disease based on serum metabolomics and network pharmacology
    LIU Guo-liang, LI Li, LEI Xia, XU Hong-dan, ZHANG Ning, YAO Yuan
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 1027-1037.   DOI: 10.16333/j.1001-6880.2023.6.012
    Abstract218)      PDF(pc) (2081KB)(4)       Save
    This present investigation uses high throughput serum metabolomics and network pharmacology techniques to explore the pharmacological mechanism of Erzhi Pill in treating Alzheimer′s disease.Sixty male SD rats were randomly divided into 6 groups according to their body weight,with 10 in each group,namely,sham operation group,model group,positive drug group,low dose group,and medium dose combined high dose group.Rats in the model group and each administration group were given intraperitoneal injection of D-galactose and bilateral intraventricular injection β amyloid protein 25-35 (Aβ25-35) to establish a rat model of Alzheimer′s disease.After the sixth week of model preparation,the medication group was given corresponding drugs for intervention every day,while the other groups were given the same volume of physiological saline (4 weeks).The spatial learning and memory abilities of rats in each group were observed using Morris water maze experiment,and the expression of tau protein in the hippocampus of rats was detected using Western blotting.Using high throughput serum metabolomics technology combined with pattern recognition methods to search for endogenous differential metabolites and related metabolic pathways;Using network pharmacology technology to explore the potential active ingredients and key targets of Erzhi Pills,further integrating metabolomics and network pharmacology technology to focus on key targets and components,and ultimately adopting molecular biology technology for targeted verification.The water maze experiment and the degree of tau protein in the hippocampus of rats proved that the model was successfully prepared.The metabolomics results showed that the metabolic profile of the model group and the sham operation group was significantly separated,and the administration group was between the sham operation group and the model group,which demonstrated that the overall metabolism of the model group rats had significant changes,and there was a trend of regression after administration of Erzhi Pill.The results of metabolomics revealed 16 biomarkers,mainly involving 10 metabolic pathways.The results of network pharmacology show that Erzhi Pill mainly contains 13 compounds including β-sitosterol,kaempferol,etc.and 11 protein components such as the toxic muscarinic acetylcholine receptor M1,a direct target associated with Alzheimer′s disease.Based on the above,it can be seen that Erzhi Pill has a good intervention effect on Alzheimer′s disease rats,which may achieve the purpose of intervention through the direct regulation of quercetin on cathepsin D.
    Related Articles | Metrics | Comments0
    Antidepressant effect of essential oil from Schizonepeta tenuifolia Briq.on lipopolysaccharide-induced depression model mice
    QIN Tian-tian, HU Jing-wen, ZENG Jiu-seng, LIU Rong, ZENG Nan
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (9): 1480-1485.   DOI: 10.16333/j.1001-6880.2023.9.002
    Abstract214)      PDF(pc) (1014KB)(41)       Save
    To investigate the antidepressant effect and possible mechanism of essential oil from Schizonepeta tenuifolia Briq.(EOST) on lipopolysaccharide (LPS)-induced depressive behaviors and NOD-like receptor pyrin domain-containing 3 (NLRP3) signaling pathway.In this study,the mice model with depression was established by intraperitoneal injection of LPS,and drug treated mice were intragastrically administered with EOST or fluoxetine.Sucrose preference test and splash test (ST) were performed at the end of the experiment.Interleukin 18 (IL-18) and tumor necrosis factor α(TNF-α) in serum were measured by enzyme-linked immunosorbent assay (ELISA),Nissl staining was used for observation of the changes of neurons in the hippocampus CA3 area of mice.The relative protein expression levels of NLRP3 inflammasome were detected by Western blot.The results showed that EOST 100,50 mg/kg significantly increased the preference of sugar water,prolong the grooming time of ST in mice,which demonstrated significant antidepressant-like behavioral effects.Additionally,it also reversed the upregulated IL-18 and TNF-α content in the serum of model mice,increased the mean IOD of Nissl bodies in the hippocampal CA3 region,down-regulated the expression levels of NLRP3,apoptosis-associated speck-like protein containing a CARD (ASC),cysteinyl aspartate specific proteinase-1 (Caspase-1),and ionized calcium binding adapter molecule 1 (Iba-1).EOST could combat the depression-like behaviors in depression model mice induced by LPS,which might be related to inhibit the activation of NLRP3 inflammasome and microglia,so as to alleviate neuroinflammation and neuronal damage.
    Related Articles | Metrics | Comments0
    Principal component analysis and comprehensive evaluation of free amino acids in mulberry leaves from different producing areas and varieties
    CHEN Shuang, YAO Huan, YUAN Lin, WU Ying-mei, LI Jin-jin, YANG Wen-wu, LIU Di-di, ZHOU Nong
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 938-948.   DOI: 10.16333/j.1001-6880.2023.6.003
    Abstract213)      PDF(pc) (992KB)(4)       Save
    To study the composition and content of free amino acids in the water extracts of three varieties from 17 different origins of mulberry leaves in Chongqing,providing theoretical basis for the development and utilization of mulberry leaves in this region.Leaves of Morus alba L.,M. cathayana Hemsl. and M. austalis Poi.from different origins were used as raw materials,the water extract was obtained by simulating mulberry leaf tea brewing method.The content of free amino acids was determined by HPLC fluorescence method and taste activity value (TAV) of 18 kinds of free amino acids was calculated.Then,the nutritional components and flavor characteristics of mulberry leaves were comprehensively evaluated by correlation analysis,principal component analysis,cluster analysis,etc.The results showed that there was no significant difference in the types of amino acids in mulberry leaves from different varieties and origins,but the content of amino acids were obvious differences.The highest content of total amino acids,essential amino acids,semi-essential amino acids and non-essential amino acids were 1 927.33 mg/100 g,322.6 mg/100 g,46.17 mg/100 g and 1 558.56 mg/100 g,respectively.Moreover,the taste characteristics of amino acids from mulberry leaves were as follows:sweet amino acids > umami amino acids > bitter amino acids.The 17 samples were divided into two categories by cluster analysis and the results showed that the samples of M. austalis Poi.(S15) and M. cathayana Hemsl.(S7) from Anlan Town in Banan District of Chongqing have high quality of amino acids.In summary,mulberry leaves are rich in amino acids and have high nutritional value,which can be used to develop amino acids functional foods.
    Related Articles | Metrics | Comments0
    Mechanism of Solanum nigrum in treatment of breast cancer based on network pharmacology,molecular docking and in vitro experimental verifcation
    WEN Xin, HU Jin-hang, CHENG Min, SONG Zhong-xing
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (10): 1782-1793.   DOI: 10.16333/j.1001-6880.2023.10.015
    Abstract211)      PDF(pc) (3022KB)(11)       Save
    To explore the mechanism of action of Solanum nigrum in the treatment of breast cancer based on network pharmacology and cellular experiments.The TCMSP database,Swiss-Target-Prediction and literature mining were used to collect seven active ingredients of S. nigrum.One hundred and ten target genes related to breast cancer were collected through the GeneCards database,and Cytoscape 3.8.0 software was used to construct the "drug-active-ingredient-target-disease" network.The potential targets were imported into the STRING 11.5 database to obtain the PPI network,and GO and KEGG enrichment analyses were performed on the potential targets using the DAVID Database.Go function enrichment analysis found 694 items.KEGG pathway enrichment analysis found 128 items (P < 0.05),involving cancer,PI3K/AKT,MAPK and other signaling pathways related to S. nigrum. Molecular docking of seven key active ingredients medioresinol,β-carotene,sitosterol,diosgenin,solanocapsine,cholesterol and quercetin from S. nigrum with potential targets AKT1,ESR1,EGFR,SRC,MAPK1,using AutoDock-vina 1.1.2 software.The molecular docking results showed that the predicted key component,diosgenin,showed good binding to the core targets,such as AKT1 and EGFR.The results showed that different concentrations of diosgenin inhibited the proliferation and promoted apoptosis of triple-negative breast cancer cells MDA-MB-231,and diosgenin regulated the expression of EGFR,AKT1 and p-AKT1 proteins,meanwhile diosgenin also down-regulated the expression of anti-apoptotic protein Bcl-2 and up-regulated the expression of pro-apoptotic protein Bax.The above results suggest that the potential mechanism of action of S. nigrum against breast cancer may be related to the regulation of MDA-MB-231 cell proliferation and apoptosis,and through two key target genes AKT1 and EGFR.
    Related Articles | Metrics | Comments0
    Evaluation on germplasm resources of genuine Amomum villosum
    TU Yu-ting, LI Ming-xiao, TANG Li-yun, SU Jing, HE Zhuo-hang, XU Jie, ZHANG Yu-yi, HE Guo-zhen
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1222-1234.   DOI: 10.16333/j.1001-6880.2023.7.014
    Abstract204)      PDF(pc) (2222KB)(9)       Save
    There are 15 towns and two streets in Yangchun city of Guangdong province which is the genuine producing area of Amomum villosum.In order to select high-yield and high-quality germplasm of A. villosum,28 A. villosum germplasms were collected from different producing areas in Yangchun City.The contents of volatile oil,bornyl acetate and chemical components of A. villosum fruit in each producing area were detected,and the germplasm resources were evaluated combining with the yield of A. villosum.The results showed that there were significant differences in yield per mu,the contents of volatile oil and bornyl acetate.A comparison of the yields of 28 A. villosum germplasm,the yields of the germplasms of XAC,XBW,XBC,XBI,XAB,XBH,XBA,XBR,XBK,XAM,XBG,XAF,XBX,XBO and XAP were greater than 40 kg per 666.67 m2,which are much higher than the average yield of A. villosum in Yangchun City,so the germplasms are defined as high-yield germplasm.The essential oil and bornyl acetate contents of the germplasms of XBR,XBC,XAP,XAN,XBF,XBM,XBD,XBH,XAL,XAU,XBE,XBV and XBG exceeded 3.50% and 2.20% respectively,which are much higher than the limit values of Pharmacopoeia of the People′s Republic of China (2020 Edition).All of 28 germplasms of A. villosum were clustered into 4 categories by analyzing the chemical components in the volatile oil of A. villosum fruits.Conclusion:there are differences in the quality of A. villosum from different places of Yangchun City.The germplasms of XBC,XBH,XBR,XBG and XAP can be the candidates of high-yielding and high-quality of A. villosum basing on their high yield exceeding 40 kg per 666.67 m2,the high essential oil content exceeding 3.50% and the high bornyl acetate content exceeding 2.20%.
    Related Articles | Metrics | Comments0
    Study on the target of julibroside J8 inhibiting the proliferation of vascular endothelial cells and its apoptosis related cell signal pathways based on molecular docking technology
    HUA Hui, SHA Xiu-xiu, JIANG Yu-chen, QIAN Xu-wu, LIU Shu
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1206-1211.   DOI: 10.16333/j.1001-6880.2023.7.012
    Abstract200)      PDF(pc) (860KB)(2)       Save
    To investigate the target of julibroside J8 (J8) inhibiting the proliferation of tumor vascular endothelial cells and its related signal pathway of apoptosis.In this paper,HPLC method was used to detect the content change of J8 in and out of endothelial cells after being treated with J8;Vina software docked J8 with VEGF,FAS,DR3,DR4,DR5,TFR-1 for analysis;Western blot method was used to detect the changes in the expression levels of VEGF,p-JNK,Bax,ENDOG,Caspase-3,Caspase-8,and Caspase-9 proteins in endothelial cells before and after drug addition.The results of molecular docking showed that the target proteins corresponding to VEGF and FAS had good binding ability to J8 and could bind to multiple sites.After HUVEC cells were treated with J8 for 24 h,the expressions of VEGF,p-JNK and other proteins were significantly decreased,and the expressions of apoptosis inducing proteins Bax and EnDOG were significantly up-regulated,and there was no significant effect on the expression levels of Caspase-3,Caspase-8 and Caspase-9.J8 may inhibit the proliferation of vascular endothelial cells by binding with VEGF on the surface of vascular endothelial cell membrane,and cause endothelial cell apoptosis by weakening the activity of VEGF/JNK pathway.
    Related Articles | Metrics | Comments0
    Study on the key components of total flavonoids from Lamiophlomis Herba in the treatment of rheumatoid arthritis based on spectrum-effect relationship and component knockout
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (11): 1833-1841.   DOI: 10.16333/j.1001-6880.2023.11.001
    Abstract199)      PDF(pc) (1153KB)(8)       Save
    This study aimed to explore the key components of total flavonoids from Lamiophlomis Herba(LH) against rheumatoid arthritis (RA),which can elucidate the active components of LH against RA.In this work,the fingerprint of 13 batches of the total flavonoids from LH was established,including eight common peaks,of which three were identified.Based on the effect of 13 batches of total flavonoids from LH on the activity of fibroblast-like synoviocytes (FLS),the spectrum-effect relationship was established for the anti-RA effect of total flavonoids from LH.The results showed that luteoloside and luteolin were key components of total flavonoids from LH against RA.Then,the component knockout technique was used to investigate the anti-RA efficacy of the luteoloside component in the total flavonoids from LH,and the effects of luteoloside and luteolin on FLS cell viability were compared.The results showed that luteoloside and luteolin could significantly inhibit FLS cell viability (P<0.01),and there was no significant difference between their inhibition rates. Because the HPLC spectrum of total flavonoids form LH showed that the content of luteoloside was higher than that of luteolin,luteoloside is a key component of total flavonoids from LH against RA.This work can provide a certain reference for the clinical application of LH in the treatment of RA.
    Related Articles | Metrics | Comments0
    Effect of pH and temperature on the property of anthocyanins from blueberry pomace in antibacterial preservation applications
    ZHAI Yue, ZHAO Hai-tao, FU Qun
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 1007-1016.   DOI: 10.16333/j.1001-6880.2023.6.010
    Abstract199)      PDF(pc) (2089KB)(13)       Save
    In this study,the colour and properties of anthocyanins from blueberry pomace were comparatively evaluated at different pH values and temperatures,with the aim of investigating the presentation of their properties in food preservation applications and promoting the multi-path and high-value utilisation of by-product pomace in the blueberry industry.The effects of pH and temperature on the colour and properties of blueberry pomace anthocyanins were investigated using content,colour,DPPH radical scavenging,hydroxyl radical scavenging,total reducing capacity,anti-lipid peroxidation capacity,inhibition circle diameter,minimum inhibitory concentration and minimum bactericidal concentration as indicators,and the inhibition mechanism was investigated using liquid conductivity,protein content and alkaline phosphatase activity as indicators.The results showed that blueberry pomace anthocyanins showed a significant colour difference from dark red to dark purple (P > 0.05) in the pH range of 2-8.Under the treatment temperature of -18-70 ℃,the content,antioxidant activity and antibacterial activity of blueberry pomace anthocyanins decreased with increasing treatment time,and the overall colour trend was blue-green.The anthocyanins of blueberry pomace changed the cell membrane permeability of the test strains and showed positive correlation with the conductivity,protein content and alkaline phosphatase activity of the bacterial solution,inhibiting the growth of the test strains.Blueberry pomace anthocyanins showed significant differences in application properties at different pH and temperature,showing strong in vitro antioxidant and antibacterial properties,which can be applied to the development of colour preservative applications.
    Related Articles | Metrics | Comments0
    Metabolomics-based analysis on the differences in antioxidant components from callus of Lycium barbarum L.
    ZENG Xiao-qian, ZHOU Xiao-ru, LIU Chun-huan, LIU Xue, YANG Cheng
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (9): 1495-1504.   DOI: 10.16333/j.1001-6880.2023.9.004
    Abstract194)      PDF(pc) (1862KB)(42)       Save
    The effects of different plant growth regulator combinations on the enrichment of antioxidant components in Lycium barbarum L. callus were investigated by non-targeted metabolomics technique.The sterile stem segments of L. barbarum was used as explant,and 0.25 mg/L naphthylacetic acid+ 0.125 mg/L 6-benzylaminopurine (NB) or 0.25 mg/L 2,4-dichlorophenoxyacetic acid+ 0.125 mg/L kinetin (DK) were used as plant growth regulators for callus culture.The antioxidant activity of different callus was detected by DPPH free radical scavenging method.The antioxidant activity of NB was 1.9 times that of DK at the same concentration.Based on the metabolomics technology of UHPLC-Q-TOF-MS,a total of 752 metabolites were identified in two groups of samples,of which 55 had significant differences in relative content,mainly amino acids and carbohydrates;KEGG analysis revealed that the differential metabolic pathway was mainly the ABC transporter pathway.The functional difference of ABC transporters may be the main reason for the significant difference in antioxidant activity of L. barbarum callus.
    Related Articles | Metrics | Comments0
    Chemical constituents from Armillaria mellea fermentation broth and its mycelia
    LUO Ying-zi, HUANG Ming-jin, SHEN Shou-mao, SU Ming-zhi, YU Dan-dan, GUO Yue-wei, QIN Yu-qiang, ZHANG Guang-wen
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1183-1190.   DOI: 10.16333/j.1001-6880.2023.7.009
    Abstract193)      PDF(pc) (509KB)(24)       Save

    Armillaria mellea is a symbiotic fungus essential for the nutritional growth stage of the medicinal plant Gastrodia elata.To investigate the chemical constitutes and bioactivities of A. mellea,the 95% ethanol extract of A. mellea was repeatedly chromatographed over silica gel,MCI gel,Sephadex LH-20 and reversed-phase (RP)-HPLC,affording twenty-six pure compounds (1-26).Their structures were identified as tetradecanoic acid (1),α-linolenic acid (2),oleic acid (3),linoleic acid (4),ethyl linoleate (5),8(R),11(S)-dihydroxy-9Z,12Z-octadecadienoic acid (6),2-linoleoylglycerol (7),1-monolinolein (8),9-octadecenoic acid-2′,3′-dihydroxy propyl ester (9),methyl linoleate (10),glycerin 1,3-dilinoleate (11),(5Z,9Z)-17-methylnonadeca-5,9-dienoate (12),(2S)-α-(9′Z,12′Z,15′Z)-octadecatrienoic acid (13),ergosta-5,7,22-triene-3β-ol (14),(22E,24S)-5α,8α-epidioxy-24-methyl-cholesta-6,9(11),22-trien-3β-ol (15),p-hydroxybenzylethyl ether (16),uracil (17),cyclo(D)-Pro-(D)-Ile (18),cyclo(D)-Pro-(D)-Leu (19),cyclo(D)-Pro-(D)-Phe (20),proline (21),cyclo(D)-Pro-(L)-Val (22),cyclo(D)-Pro-(D)-Val (23),4-(2-hydroxyethyl)-5-methyloxazole (24),cyclo-(Ala-Val) (25),cyclo-(Ala-Pro) (26),respectively,by comparision of their NMR data and optical rotation values with those reported in the literature. Among them,compounds 5-13,16,and 18-26 were obtained for the first time from A. mellea. In in vitro bioassay,compounds 14 and 15 showed moderate effects against LPS-induced inflammatory responses in RAW 264.7 cells.

    Related Articles | Metrics | Comments0
    Optimization of extraction technology of alcohol extract from Achyranthes aspera L.and its activity against human lung adenocarcinoma A549 cells in vitro
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1191-1197.   DOI: 10.16333/j.1001-6880.2023.7.010
    Abstract182)      PDF(pc) (1226KB)(2)       Save
    Achyranthes aspera L. is a widely used medicinal plant with many effects.In order to study the anti-cancer effect of its alcohol extract,the alcohol extraction process from A. aspera was optimized by Box-Behnken response surface method,and the cytotoxicity and cell scratch test of human lung cancer A549 cells were carried out in vitro.The optimum extraction conditions were as follows:extraction temperature 90 ℃,liquid-solid ratio 40∶1,extraction time 2 h.Under these conditions,the extraction rate of alcohol extract from A. aspera was 9.23%.The results of cytotoxicity test showed that the survival rate of A549 cells decreased rapidly with the increase of alcohol extract concentration.When the concentration of alcohol extract was 256 μg/mL,the inhibition rate of A549 cells was 94.65%.The cell scratch results showed that alcohol extract had a certain inhibitory effect on the migration of A549 cells,which can provide a reference for the study of the medicinal properties from A. aspera.
    Related Articles | Metrics | Comments0
    Effect of luteolin on Th17/Treg immune balance in mice with ulcerative colitis by regulating SIRT3/AMPK/mTOR signaling pathway
    YUAN Li, JI Jian-hua, LI Min-yan
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (7): 1144-1153.   DOI: 10.16333/j.1001-6880.2023.7.005
    Abstract180)      PDF(pc) (1752KB)(5)       Save
    To explore the effect of luteolin on the immune balance of T helper 17 cells (Th17)/regulatory T cells (Treg) in a mouse model of ulcerative colitis (UC),and to analyze its underlying mechanism.Sixty BALB/c male mice were randomly separated into normal group,model group,positive control group,luteolin group,luteolin+3-TYP group,with 12 mice in each group.Dextran sodium sulfate (DSS) was performed to induce the establishment of a UC model,the weight change,stool consistency and stool occult blood in mice were observed and recorded,and the disease activity index (DAI) was calculated;HE staining was used to observe the histopathological changes of the colon;flow cytometry was used to measure the ratio of Th17 and Treg cells in the spleen,and the ratio of Th17/Treg was calculated;ELISA was used to measure the contents of IL-6,IL-10,IL-17 and IL-23 in colon tissue;RT-qPCR was used to measure the mRNA expression of RORγt and Foxp3 in colon tissue;Western blot was used to measure the expression of colonic tissue SIRT3,AMPK,p-AMPK,mTOR,p-mTOR proteins.Compared with the normal group,the DAI score,histopathology score,spleen Th17/Treg ratio,colon tissue IL-6,IL-17,IL-23,RORγt mRNA levels and p-mTOR/mTOR ratio of mice increased in the model group,the IL-10,Foxp3 mRNA and SIRT3 protein levels,and p-AMPK/AMPK ratio decreased (P<0.05).After drug intervention,the DAI score,histopathology score,spleen Th17/Treg ratio,colon tissue IL-6,IL-17,IL-23,RORγt mRNA levels and p-mTOR/mTOR ratio of mice decreased,the IL-10,Foxp3 mRNA and SIRT3 protein levels,and p-AMPK/AMPK ratio increased P<0.05);3-TYP was able to weaken the effect of luteolin on the differentiation balance of Th17/Treg cells in UC mice (P<0.05).Luteolin can improve Th17/Treg imbalance in DSS-induced UC mouse model,and its mechanism may be related to the activation of SIRT3/AMPK/mTOR pathway.
    Related Articles | Metrics | Comments0
    Banana peel extract improves intestinal motility of constipation model mice by regulating intestinal flora and intestinal neurotransmitter levels
    SHEN Yu-ping, HU Qing, SUN Li-jun, LIAO Jian-meng, FANG Zhi-jia, DENG Qi, SUN Dong-fang, WANG Chen
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (5): 750-760.   DOI: 10.16333/j.1001-6880.2023.5.003
    Abstract178)      PDF(pc) (2751KB)(10)       Save
    To explore the effect of banana peel extract on intestinal motility in constipated mice.In this study,the mice model of constipation was established with the mixed antibiotic solution.The effects of banana peel extract on intestinal flora,intestinal neurotransmitter and intestinal motiveness of constipated mice were measured.The results showed that the fecal water content increased by 137.5%,the intestinal propulsion rate increased by 3.4 times,and the first black stool excretion time decreased by 73.7% (P< 0.05) in the high-concentration banana peel extract group,which basically recovered to the level of the blank group.The phyla horizontal flora was disordered and some beneficial bacteria were missing in the constipation model mice.After the intervention of low concentration banana peel extract,the relative abundance of Trabulsiella and Unclassified-f-Enterobacteriaceae increased by 6 times,the relative abundance of intestinal Bacteroides decreased by 94.4%.Furthermore,the concentrations of norepinephrine,dopamine,gamma-aminobutyric acid and acetylcholine of neurotransmitters were significantly increased by 6.1%,17.6%,63.7% and 52.6%.These results indicated that banana peel extract could regulate intestinal flora and intestinal neurotransmitter levels to improve intestinal motility in constipated mice.
    Related Articles | Metrics | Comments0
    Biological activity of tangeretin and its potential efficacy in exercise-induced asthma:a review
    LIU Meng, ZHANG Zheng, GAO Bing-hong, ZHOU Zhi-qin
    NATURAL PRODUCT RESEARCH AND DEVELOPMENT    2023, 35 (6): 1088-1099.   DOI: 10.16333/j.1001-6880.2023.6.018
    Abstract175)      PDF(pc) (1768KB)(10)       Save
    Exercise-induced asthma (EIA) is one of the most frequent chronic diseases among athletes and is gaining increasing concern.The pathological mechanisms and pharmacology-based treatments of EIA have been widely studied and some progress has been made,however,long-term using EIA drugs would induce health-related side effects and even doping violations.Thus,a safe,healthy and effective alternative of traditional treatment methods is particularly required.Tangeretin extracted from a natural citrus was recently reported to show great promise in EIA treatment.Therefore,the current article specifically reviewed the biological activities of tangeretin referring to anti-inflammatory,antioxidant,anti-cancer,regulation of uric acid and cortisol secretion,regulation of glucolipid metabolism and anti-obesity,neuroprotection.It would provide ideas and theoretical support for further studying and developing tangeretin-based food,healthcare products or drugs in EIA treatment.
    Related Articles | Metrics | Comments0