1.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.
2.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.
3."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.
4.Differentiation on Original Plants of Xi-Huang-Cao through DNA Barcodes
Huiye ZHANG ; Feng LIU ; Deqin WANG ; Ruoting ZHAN ; Xinye MA ; Weiwen CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(7):1487-1490
This study was aimed to identify original plants of Xi-Huang-Cao (XHC) through DNA barcodes. The nu-cleotide sequence information of rbcL, psbA-trnH, matK and ITS2 regions were abstracted using standardized man-ners from 41 samples (including Rabdosia serra, R. lophanthoides and R. lophanthoides var. graciliflora). Sequencing efficiency of each marker was calculated. Species identification capability was tested on the basis of TaxonGap. The results showed that sequence success rates were 100% for rbcL, 90.2% for psbA-trnH, 87.8% for ITS2, and 70.7%for matK. Three markers (rbcL, psbA-trnH and ITS2) were competent for species discrimination (not for subspecies). It was concluded that rbcL can be the preferred barcode for XHC because of its convenience and efficacy.
5.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.
6.Models of adenoviral transfection and hypoxia/reoxygenation-induced injury in AMCMs of adult mouse cardiomyocytes
Xiaoru LI ; Xinye YAO ; Jia LIU ; Xiaoyu ZHANG ; Yiman ZHANG ; Baochang LAI ; Qiang MA ; Yidong WANG ; Hongyan TIAN ; Qian YIN
Acta Laboratorium Animalis Scientia Sinica 2024;32(4):435-443
Objective To construct models of viral transfection and hypoxia/reoxygenation induced cellular injury in adult mouse cardiomyocytes(AMCMs)isolated using a non-Langendorff method.Methods AMCMs were isolated,extracted,sedimented,and plated using a non-Langendorff method.The morphology and survival rate of the isolated cells were evaluated 2,24,48 and 72 h after plating,and their integrity was observed by immunofluorescence staining for α-actinin.The isolated AMCMs were infected with adenoviruses carrying an RFP-expressing vector and fluorescence images were obtained at 36 and 48 h post-infection and used to calculate transfection efficiency.The cells were cultured under hypoxic conditions for 45 min,reoxygenated for 24 h,and then stained with propidium iodide(PI)to verify establishment of the hypoxia/reoxygenation injury model.Results The survival rates of AMCMs at 2,24 and 48 h after plating were comparable,but survival was significantly reduced at 72 h.The integrity of the AMCMs was good and>80%of the cells were transfected with adenovirus at 48 h.After hypoxia/reoxygenation treatment,42%of cells were stained by PI,suggesting successful establishment of the AMCM injury model.Conclusions In this study,we developed a non-Langendorff method for the fast and easy isolation of AMCMs with high cell viability.The isolated cells can be efficiently infected with adenovirus and respond to hypoxia/reoxygenation injury.These findings provide a systematic method for isolating AMCMs and for applying gene modification and hypoxia/reoxygenation injury in these cells.