1.Experimental Study of Anti-inflammation of Durian Peel Extract
Guo XIE ; Minzhi WU ; Jinle CHENG ; Ruoting ZHAN ; Weiwen CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(1):130-135
Objective To investigate the anti-inflammatory effects and mechanism of durian peel extracts (DPE). Methods The in vivo anti-inflammation effects of DPE were examined by carrageenin-induced mice paw edema test and allergic contact dermatitis test induced by 2, 4-DNFB. And the in vitro anti-inflammation effects of DPE were evaluated with methyl thiazolyl tetrazolium ( MTT) assay in RAW 264.7 cell model of inflammation induced by lipopolysaccharide (LPS). Results The results of animal experiments showed that DPE groups could markedly relieve mice paw edema induced by carrageenin ( P<0.01 or P<0.001 compared with blank group). DPE could effectively inhibit the allergic contact dermatitis induced by 2, 4-DNFB in mice, showing good dose-effect relationship. The results of in-vitro test showed that DPE at the given concentrations had no influence on RAW 264.7 cell proliferation. Tumor necrosis factor alpha ( TNF-α) , interleukin 6 ( IL-6) , interleukin 1 beta (IL-1β), nitric oxide (NO) and nuclear factor kappaB (NF-кВ ) were observably inhibited, and anti-inflammatory cytokine IL-10 was enhanced by 25 and 50 mg/L of DPE. Conclusion DPE exert potential anti-inflammation effect, and the mechanism might be related to its inhibition of NF-кВsignal pathway.
2.In-vitro Antibacterial Activity of Herba Taraxaci Extract on Escherichia coli
Xiaoyu JI ; Yuanxia PENG ; Min LIU ; Hui XU ; Ruoting ZHAN
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(1):116-120
Objective To evaluate the inhibitory activity of Herba Taraxaci extract on Escherichia coli DH5α (E. coli DH5α) and to investigate proteomic response of E. coli. Methods Medicinal powder of Herba Taraxaci was extracted with the solvents of different polarity ( n-hexane, ethyl acetate, distilled water) , and then the obtained 8 different extracts were subjected to thin layer chromatography ( TLC) analysis. Microdilution method was performed to detect the minimum inhibitory concentration ( MIC) of different extracts and the growth curves were described. The protein expression profiles of E . coli treated with the extracts were analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis ( SDS-PAGE) and two dimensional electrophoresis (2-DE) . Results Water decoction of Herba Taraxaci could obviously suppress the growth of E. coli with a MIC of 1.95 mg/mL. The different extractions exhibited no antibacterial activity except ethyl acetate phase 3 with a MIC of 0.13 mg/mL, which was equal to 19.23 mg/mL of crude drugs. The results of TLC analysis showed that chlorogenic acid was undetectable in n-hexane extract and ethyl acetate phase 1 extract, and ethyl acetate phase 2 and 3 extracts showed obviously increased spots. The results of SDS-PAGE and 2-DE showed that water decoction of Herba Taraxaci had inhibitory effect on the expression of functional protein. The results of 2-DE showed that after treatment with ethyl acetate phase 3 at the concentration of 2 × MIC for 21 hours, the amount of protein spots were 92 less than those of the blank control group, the spots of E. coli DH5α soluble protein with expression amount down-regulated doubly were 24, and those with expression amount up-regulated doubly were 19. Ethyl acetate phase 3 extract had an effect on down-regulating the protein expression of E. coli DH5α soluble protein pH3-10, and water decoction of Herba Taraxaci had inhibitory effect on E. coli DH5αprotein expression. Conclusion Herba Taraxaci has significant antibacterial activity on E. coli DH5α, and the water-soluble fraction of chlorogenic acid and caffeic acid might be the active components. The possible antibacterial mechanism may be related with the regulation of bacterial protein expression.
3.Study on Callus Induction in Explant of Aquiliaria sinensis Root
Yanfengyang JIANG ; Chao LIN ; Ruoting ZHAN ; Xinye MA
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(4):751-754
Objective To study the conditions of callus induction with the roots of Aquilaria sinensis as explants. Methods Two sources of roots of Aquiliaria sinensis were selected as the explants. The effects of sterilization methods and the combination of different concentrations of phytohormones on callus induction were evaluated. Results When Aquiliaria sinensis root seedling was sterilized in 0.01mg/mL HgCl2 solution for 3 minutes, the sterilized effect was the best. The optimal callus induction medium was MS+0.1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) +0.1 mg/L 6-benzylaminopurine (6-BA). Aseptic Aquiliaria sinensis root seedling cultivated in callus induction medium containing MS+1.0 mg/L naphthalene acetic acid ( NAA) +0.8 mg/L 6-BA achieved the highest callus induction rate. Conclusion Callus can be induced from two sources of Aquilaria sinensis roots. The induction rate of callus is lower when the explant root seedling is cultivated using 2,4-D alone as inducer, and is increased when used together with 6-BA.
4.Identification of Original Plants of Guanzhong (Dryopteris Species) by Chloroplast psbA-trnH Intergenic Region
Feng LIU ; Yanfengyang JIANG ; Ruoting ZHAN ; Xinye MA ; Weiwen CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(7):1482-1486
This article was aimed to study the species identification capability of psbA-trnH among Guanzhong herbs from Dryopteri. The nucleotide sequence information of psbA-trnH region was abstracted using standardized manners from 9 Dryopteri species (19 samples). The identification efficiency of psbA-trnH was analyzed based on TaxonGap among both tested materials (9 species, 19 samples) and tested materials plus GenBank data (17 species, 44 samples in total). The results showed that with the expanded species range, the discriminative efficiency of psbA-trnH de-clined from 100% (9/9) to 82.4% (14/17), while the proportion of within-specific heterogeneity larger than between-specific separability increased from 11.1% (1/9) to 52.9% (9/17). It was concluded that psbA-trnH can be recom-mended as the valuable barcode for homonym distinguishment among Guanzhong herbs, with attention of adequate sampling within species.
5.Transcriptomic Analysis and Systematic Mining of Genes Involved in Biosynthetic Pathway of Triterpenoid Saponins in Ilex Asprella
Xiasheng ZHENG ; Xiuxiu LUO ; Hui XU ; Ruoting ZHAN ; Weiwen CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(7):1505-1512
This study was aimed to screen candidate genes involved in the triterpenoid saponins biosynthetic pathway of the Ilex asprella root. The Illumina platform was applied to perform transcriptomic sequencing of I. asprella root, followed by a series of bioinformatics analysis. The results showed that a total of 272 candidate unigenes were anno-tated to be involved in the biosynthetic pathway of terpenoid in the transcriptome of I. asprella root, including 72 u-nigenes for the upstream pathway and 26 unigenes for cyclization, oxidation and glycosylation in the downstream pathway. Phylogenetic analysis was carried out to further analyze the evolution relationship of some candidate uni-genes and their homologous genes. Two genes IaA S1 and IaA S2 were proved to be mixed amyrin synthases in yeast expression system. Moreover, IaA S1 was identified to one of the rare ASs with α-amyrin as the major product. It was concluded that a series of candidate genes, which might be involved in the biosynthetic pathway of triterpenoid saponins, were screened out from the transcriptome of I. asprella root. Further investigation of these candidate genes will provide insight into their actual functions in the triterpenoid saponins biosynthetic pathway in I. asprella.
6.The empirical research on cultivation mechanism of manufacture-learning-research cooperation for postgraduates of science of Chinese Pharmacology of Guangzhou University of Chinese Medicine
Huafeng PAN ; Sichen WU ; Xin GE ; Jinling REN ; Yichong FENG ; Ruoting ZHAN
Chinese Journal of Medical Education Research 2011;10(10):1176-1178
The paper summarized the successful experience such as preliminarily establishing cultivation mechanism of manufacture-learning-research cooperation for postgraduates of science of Chinese pharmacology of Guangzhou University of Chinese Medicine,ensuring innovation ability training for postgraduates,getting reasonable configuration for social education resources,and put forward existing problems in traditional Chinese medicine system,such as the loose management and faulty evalution system.Suggestion is to futher perfect relevant rules and regulations,clear and definite responsibility of school and enterprise,reserch regularly to impove ideological education of postgraduates,etc.In the way,we can achieve the aim of promoting high-level talents of traditional Chinese medicine industry.
7."Opinions on a New Cross-discipline ""Biological Chemistry of Chinese Herbal Germplasm Resources"""
Weiwen CHEN ; Hui XU ; Ruoting ZHAN ; Jinfen YANG ; Rui HE ; Jiawei LIU ; Ping YAN ; Xinye MA
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(4):704-709
Biological chemistry of Chinese herbal germplasm resources (BCCHGR) is a new cross-discipline formed from rapid development of modern science and technology and its application in the area of Chinese herbal resources. BCCHGR was defined as probing and understanding biological processes like heredity, gene transcription, expression and metabolism of Chinese herbal germplasm, at the interface of biochemistry, molecular biology and chemistry, elu-cidating the nature of Chinese herbal germplasm using as TCM medicine as well as the forming mechanism thereof. In this paper, the scientific background, definition, significance and contents of BCCHGR were discussed to depict a preliminary picture of BCCHGR and arouse popular consideration and discussions.
8.Molecular Identification of Amomum villosum Lour.Based on Sequence Analysis of 26S rDNA D1-D3 Region and matK Gene
Qionglin HUANG ; Jinfen YANG ; Zhonggang DUAN ; Rui HE ; Ruoting ZHAN ; Honghua XU ; Hui XU ; Weiwen CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 1999;0(02):-
Objective To establish a molecular identification method for three cultivars of Amomum villosum Lour.(AVL),thus to provide scientific evidence for the identification,selection and breeding of AVL.Methods The fragments of 26S rDNA D1-D3 region and matK gene of three cultivars of AVL and Amomum longiligulare T.L.Wu were amplified by polymerase chain reaction(PCR) and with corresponding primers,and then their sequences were analyzed,and phylogenetic tree was constructed based on the sequences.Results We obtained 739 bp in 26S rDNA D1-D3 sequence.Differences in 4 basic sites of 739 bp were shown between AVL and Amomum longiligulare T.L.Wu.The two cultivars of AVL,Changguo and Yuanguo,had the same sequence,but there was a difference in one basic site of Changguo and Yuanguo from Chunxuan.The phylogenetic tree based on 26S rDNA D1-D3 sequence revealed the difference between Chunxuan and the other two cultivars of AVL.We also obtained 824 bp in matK gene sequence.The three cultivars of AVL showed the consistent sequence,but there was a difference in one basic site of three cultivars of AVL from Amomum longiligulare T.L.Wu.Conclusion We can identify the three cultivars of AVL through the sequence differences at the molecular level,and Chunxuan has a closer genetic relationship with Amomum longiligulare T.L.Wu.
9.Studies on Quality Standard of Radix Toddaliae Asiaticae
Lingling WEN ; Runsheng ZHENG ; Yaping XU ; Zehui QIN ; Hui XU ; Ruoting ZHAN ; Weiwen CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(1):136-140,146
Objective To establish the quality standard of Radix Toddaliae Asiaticae. Methods Thin layer chromatography ( TLC) and high performance liquid chromatography ( HPLC) were used to identify and determine chloride nitidine and toddalolactone in Radix Toddaliae Asiaticae. The moisture and total ash contents were detected according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition) . Results Toddalolactone and chloride nitidine were detectable by TLC, the spots were clear and the dissociation was good. The established HPLC method was simple and accurate. The linear ranges of toddalolactone and chloride nitidine in Radix Toddaliae Asiaticae were 2.84~42.6 μg/mL and 25.6~385 μg/mL, and their recovery rates were 99.2 % ( RSD=1.12%) and 100 % ( RSD=0.71%) , respectively. The content of moisture was in the range of 75.8~98.9 mg/g and that of total ash was in the range of 12.4~33.6 mg/g. Conclusion The developed method is specific and accurate, and can provide useful reference for establishing quality standard of Radix Toddaliae Asiaticae.
10.Quality Evaluation of Lignum Dalbergiae Odoriferae Medicinal Materials
Yalian WANG ; Ping YAN ; Yi ZHOU ; Tong ZHAO ; Xinye MA ; Ruoting ZHAN
Journal of Guangzhou University of Traditional Chinese Medicine 2015;32(6):1087-1090,1146
Objective To evaluate the quality of medicinal materials of Lignum Dalbergiae Odoriferae in Chinese herbal medicine market. Methods Eighteen batches of commercial medicinal materials of Lignum Dalbergiae Odoriferae were identified and analyzed by macroscopical identification, thin layer chromatography (TLC), and volatile oil assay according to the Chinese Pharmacopoeia published in 2010. Visible spectrometry was used to determine the content of total flavonoids from the qualified samples. And the gas chromatography was applied to evaluate the content of nerolidol from volatile oils of the qualified samples. Results Only 33.3%of the samples met the standard of Chinese Pharmacopoeia. The total flavonoid content of the qualified samples was in the range of 21.6-29.0 mg/g. The content of nerolidol from volatile oils of the qualified samples was in the range of 294-574 mg/g. Conclusion At present, the quality of medicinal materials of Lignum Dalbergiae Odoriferae in Chinese herbal medicine market and in clinic varies greatly, and adulterants and inferior are common. The contents of chemical components in different batches of samples are significant different.