2.Molecular identification of Manis pentadactyla using DNA barcoding.
Jing JIA ; Hong-Yin ZHANG ; Jun CHEN ; Dong LIU ; Hui YAO ; Qi-Ni QIAN ; Hui ZHANG
China Journal of Chinese Materia Medica 2014;39(12):2212-2215
The COI gene as DNA barcode was used to identify the Manis pentadactyla and its adulterants in order to provide a scientific basis for the molecular identification of M. pentadactyla. Genomic DNA was extracted from experimental samples using the DNA extraction kit. The COI genes were amplified using polymerase chain reaction (PCR) and sequenced bi-directionally. Obtained sequences were assembled using the CodonCode Aligner. The neighbor-joining (NJ) tree was constructed by MEGA 6.0. The results indicated that COI sequences were successfully amplified and NJ trees results indicated that M. pentadactyla and its adulterants can be easily identification. Therefore, the COI gene is an efficient barcode for identification of M. pentadactyla and its adulterants,which will provide a new technique for the market supervision.
Animals
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Cattle
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DNA Barcoding, Taxonomic
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methods
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Drug Contamination
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prevention & control
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Electron Transport Complex IV
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genetics
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Mammals
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classification
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genetics
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Medicine, Chinese Traditional
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Molecular Sequence Data
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Phylogeny
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Quality Control
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Sheep
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Swine
3.Molecular Identification of Serpentis Periostracum and Its Adulterants Based on COI Sequence
Linchun SHI ; Jun CHEN ; Dong LIU ; Hongyin ZHANG ; Jing JIA ; Hui ZHANG ; Hui YAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(2):284-287
Objective: This study aimed to distinguish Serpentis Periostracum from its adulterants, which will provide the basis for its safe application. Methods: Here, COI sequences of 68 samples from 13 species were PCR amplified and sequenced. Furthermore, the DNA Barcoding Gap and phylogenetic cluster analysis were carried out. Results:The results exhibited that the COI sequences of all the three origin animals of Serpentis Periostracum have DNA Barcoding Gap. For phylogenetic cluster analysis, all the three origin animals showed monophyletic and every species can be discriminated clearly. Conclusion: COI is an effective DNA barcode for the identification of Serpen-tis Periostracum.
4.Comparative study between MRI and echocardiography in noncompaction of ventricular myocardium
Zi-Yan SUN ; Li-Ming XIA ; Cheng-Yuan WANG ; Jing-Jing YAO ; Wei-Hui SHENTU ;
Chinese Journal of Radiology 2001;0(08):-
Objective To investigate the MRI and echocardiography manifestations of noncompaction of ventricular myocardium(NVM)and assess the role of MRI in the diagnosis of NVM by comparing it with echocardiography.Methods Fourteen cases of NVM diagnosed by echocardiography were examined with MRI,including scanning of black-blood sequences,double inversion recovery fast spin echo (DIRFSE)and triple inversion recovery fast spin echo(TIRFSE),and white blood sequence:fast imaging employ steady state acquisition(FIESTA).Scanning plane includes short axis view,four-chamber view and long axis view.Results Both MRI and echocardiography displayed involvement of left ventricles in thirteen cases and involvement of double ventricles in one case.Apexes of heart and the intermedius are commonly affected.MRI showed 54 segments and echocardiography showed 53 segments affected,and there is no significant difference between the capability of MRI and echocardiography(P=1.000).The affected myocardium consisted of two layers:subendoeardial noncompacted myocardium and epicardial compacted myocardium,and the ratio measurement of N/C by MRI was 3.37?0.89 and it was 3.19?0.82 by echocardiography.Noncompacted myocardium was characterized by prominent and excessive myocardial trabeculations and deep intratrabecular recesses,in which the blood flow was communicated with the ventricle.One case was complicated with ventricular aneurysm,and coronary arteriography was performed with unremarkable findings.One case underwent heart transplantation because of progressive heart failure, Gross findings demonstrated prominent muscular trabeculations with deep intratrabecular recesses,which coincided well with MRI findings.Conclusion The MRI manifestation of NVM is characteristic,and MRI with multiple series and planes is helpful in the diagnose of NVM.Compared with echoeardiography,MRI could display the pathological cardiac muscle more clearly,because of its high soft-tissue resolution and spatial resolution.
5.Identification of Scolopendra subspinipes mutilans and its adulterants using DNA barcode.
Hong-Yin ZHANG ; Jun CHEN ; Jing JIA ; Dong LIU ; Lin-Chun SHI ; Hui ZHANG ; Jing-Yuan SONG ; Hui YAO
China Journal of Chinese Materia Medica 2014;39(12):2208-2211
In this study, the COI barcode was used to identify the Scolopendra medicinal materials and its adulterants in order to provide a new method for the identification of Scolopendra. Genomic DNA was extracted from the experimental samples. The COI sequences were amplified and sequenced bi-directionally. Sequence alignment and NJ tree construction was carried out by MEGA6.0 software. The results showed that the COI sequences can be obtained from all experimental samples. The average inter-specific K2P distance of Scolopendra was 0.222 and the minimum inter-specific distance was 0.190. All the Scolopendra subspinipes mutilans medicinal samples clustered into a clade in the NJ tree and can be distinguished from its adulterants. In a conclusion, COI can be used to correctly identify Scolopendra medicinal materials, and it will be a potential DNA barcode for identifying other animal medicinal materials.
Animals
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Arthropod Proteins
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genetics
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DNA Barcoding, Taxonomic
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methods
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Drug Contamination
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prevention & control
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Electron Transport Complex IV
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genetics
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Medicine, Chinese Traditional
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Molecular Sequence Data
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Phylogeny
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Quality Control
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Scorpions
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classification
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enzymology
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genetics
6.Identification of medicinal plant Dendrobium based on the chloroplast psbK-psbI intergenic spacer.
Hui YAO ; Pei YANG ; Hong ZHOU ; Shuang-jiao MA ; Jing-yuan SONG ; Shi-lin CHEN
Acta Pharmaceutica Sinica 2015;50(6):783-787
In this paper, the chloroplast psbK-psbI intergenic spacers of 18 species of Dendrobium and their adulterants were amplified and sequenced, and then the sequence characteristics were analyzed. The sequence lengths of chloroplast psbK-psbI regions of Dendrobium ranged from 474 to 513 bp and the GC contents were 25.4%-27.6%. The variable sites were 71 while the informative sites were 46. The inter-specific genetic distances calculated by Kimura 2-parameter (K2P) of Dendrobium were 0.006 1-0.058 1, with an average of 0.028 4. The K2P genetic distances between Dendrobium species and Bulbophyllum odoratissimum were 0.093 2-0.120 4. The NJ tree showed that the Dendrobium species can be easily differentiated from each other and 6 samples of the inspected Dendrobium species were identified successfully through sequencing the psbK-psbI intergenic spacer. Therefore, the chloroplast psbK-psbI intergenic spacer can be used as a candidate marker to identify Dendrobium species and its adulterants.
Chloroplasts
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DNA, Chloroplast
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genetics
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DNA, Plant
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genetics
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DNA, Ribosomal Spacer
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genetics
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Dendrobium
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classification
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genetics
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Plants, Medicinal
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classification
;
genetics
7.DNA barcoding identification between arisaematis rhizoma and its adulterants based on ITS2 sequences.
Lin-Chun SHI ; Jun CHEN ; Li XIANG ; Jing-Yuan SONG ; Hui YAO
China Journal of Chinese Materia Medica 2014;39(12):2176-2179
Fifty-eight samples belonging to 7 species of Arisaematis Rhizoma and its adulterants were collected. The ITS2 locus was employed as a DNA barcode and amplified, sequenced and assembled for all of the collected samples. Then, ITS2 sequences have been annotated using HMM-based method. The intra- and inter-specific variations were calculated and NJ tree was constructed using MEGA 6.0 software. The results showed that inter-specific K2P distances were significantly larger than intra-specific distances for all of the three origin species of Arisaematis Rhizoma. Furthermore, three origin species, Arisaema amurense, A. erubescens and A. heterophyllum, can be respectively formed to be a single branch with high bootstrap values. It is concluded that ITS2 can be used to correctly identify Arisaematis Rhizoma from its adulterants and the application of ITS2 in the identification of traditional Chinese medicine has an important prospective.
Arisaema
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classification
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genetics
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DNA Barcoding, Taxonomic
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methods
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DNA, Plant
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genetics
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DNA, Ribosomal Spacer
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genetics
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Drug Contamination
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prevention & control
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Drugs, Chinese Herbal
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chemistry
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classification
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Molecular Sequence Data
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Phylogeny
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Quality Control
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Rhizome
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classification
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genetics
8.Molecular identification of aucklandiae radix, vladimiriae radix, inulae radix, aristolochiae radix and kadsurae radix using ITS2 barcode.
Xiao-Chong MA ; Hui YAO ; Lan WU ; Li XIANG ; Xiao-Chen CHEN ; Jing-Yuan SONG
China Journal of Chinese Materia Medica 2014;39(12):2169-2175
In order to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix using ITS2 barcodes, genomic DNA from sixty samples was extracted and the ITS2 (internal transcribed spacer) regions were amplified and sequenced. The genetic distances were computed using MEGA 5.0 in accordance with the kimura 2-parameter (K2P) model and the neighbor-joining (NJ) phylogenetic tree was constructed. The results indicated that for Aucklandiae Radix (Aucklandia lappa), Vladimiriae Radix (Vladimiria souliei and V. souliei var. cinerea), Inulae Radix (Inula helenium), Aristolochiae Radix (Aristolochia debilis) and Kadsurae Radix (Kadsura longipedunculata), the intra-specific variation was smaller than inter-specific one. There are 162 variable sites among 272 bp after alignment of all ITS2 sequence haplotypes. For each species, the intra-specific genetic distances were also smaller than inter-specific one. Furthermore, the NJ tree strongly supported that Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix can be differentiated. At the same time, V. souliei (Dolomiaea souliei) and V. souliei var. cinerea( D. souliei var. cinerea) belonging to Vladimiriae Radix were clearly identified. In conclusion, ITS2 barcode could be used to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix. Our study may provide a scientific foundation for clinical safe use of the traditional Chinese medicines.
Aristolochia
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classification
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genetics
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Base Sequence
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DNA Barcoding, Taxonomic
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methods
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DNA, Plant
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genetics
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DNA, Ribosomal Spacer
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genetics
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Drugs, Chinese Herbal
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chemistry
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classification
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Molecular Sequence Data
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Phylogeny
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Plants, Medicinal
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classification
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genetics
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Quality Control
9.Making plan for national significant science and technology project of TCM: example of research in teaching and inheritance of experience of famous and veteran doctors of TCM
Hui LI ; Xingdong HE ; Weiliang WENG ; Naili YAO ; Jing MA ; Mengsheng CHEN
Chinese Journal of Medical Science Research Management 2008;21(4):224-226,234
In our study,we adopted a schedule plan management of the Project Management Body of Knowledge(PMBOK)in the management practice of the experimental research on "The Research in Teaching and Inheritance of Experience of Famous and Veteran Doctors of TCM " which was supported by "tenth five years" plan of national science and technology as a key program.We tried to establish a project planning based on the special features of TCM and the characters of the project.This planning played a significant support role in the top project design and ensuring the smooth implementation of the research.
10.Molecular identification of Cynomorii herba using ITS2 DNA barcoding.
Dian-Yun HOU ; Jing-Yuan SONG ; Lin-Chun SHI ; Pei YANG ; Shi-Lin CHEN ; Hui YAO
China Journal of Chinese Materia Medica 2013;38(23):4028-4032
OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.
METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.
RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.
CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.
Cynomorium ; classification ; genetics ; DNA Barcoding, Taxonomic ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Polymerase Chain Reaction