1. Molecular Identification of Polygonum chinensis Using DNA Barcoding
Chinese Pharmaceutical Journal 2018;53(14):1165-1169
OBJECTIVE: To identify Polygonum chinensis and its adulterants by ITS2 sequences. METHODS: Total genomic DNA of P. chinensis was extracted using the plant genomic DNA kit. The internal transcribed spacer 2(ITS2) regions were amplified. The variable site of ITS2 regions were analysed through MEGA 6.0 software. The intra-versus inter-specific genetic distances of the ITS2 regions was calculated based on the kimura 2-parameter(K2P) model. Neighbor-Joining phylogenetic trees were constructed using MEGA6.0. RESULTS: The intraspecific variation of P. chinensis was small. However, the interspecific variation of P. chinensis and its adulterants was small distinct. The secondary structure of ITS2 of P. chinensis and its adulterants has significant difference. NJ trees can identify P. chinensis and its adulterants. CONCLUSION: ITS2 Regions can be used to authenticate P. chinensis and its adulterants which provide new METHODS for the identification of P. chinensis. The standard DNA barcodes of P. chinensis are establishment which would lay the foundation of identification and safety clinical drug application of P. chinensis.
2. Molecular identification of Dalbergiae Odoriferae lignum and its adulterants using the ITS2 sequence
Chinese Pharmaceutical Journal 2015;50(15):1273-1276
OBJECTIVE: To identify Dalbergiae Odoriferae Lignum and its adulterants using DNA barcoding. METHODS: ITS2 is one of the popular DNA barcoding in the identification of traditional Chinese medicine. In this paper, the ITS2 regions of Dalbergiae Odoriferae Lignum and its adulterants were amplified and sequenced bi-directional. The length and GC content of ITS2 sequence were analyzed through MEGA5.0 software. The genetic distances were computed by kimura 2-parameter (K2P) model. Dalbergiae Odoriferae Lignum and its adulterants have been identified through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) phylogenetic tree. RESULTS: The sequence lengths of ITS2 of Dalbergiae Odoriferae Lignum were 216 bp, and the GC content was 68.5%. The minimum K2P interspecific genetic distances of Dalbergiae Odoriferae Lignum and its adulterants were 0.009, which was larger than that of the intraspecific genetic distances of Dalbergiae Odoriferae Lignum. The Dalbergiae Odoriferae Lignum and its adulterants can be obviously identified using the Species identification System and NJ phylogenetic trees. CONCLUSION: ITS2 Regions as DNA barcode can identify Dalbergiae Odoriferae Lignum and its adulterants accurately.
3. Application study of ITS/ITS2 barcodes in identification of Zanthoxmli Pericarpium
Chinese Pharmaceutical Journal 2014;49(7):534-538
OBJECTIVE: To evaluate the accuracy and stability of ITS/ITS2 barcodes in identification of Zanthoxmli Pericarpium. METHODS: Total genomic DNAs from samples were extracted by using improved DNA extraction kits. ITS regions were amplified, and the ITS2 sequences were obtained by using the hidden Markov model (HMM) based on annotation method from the ITS sequences. The inter- and intra-specific variation of the Zanthoxmli Pericarpium and its adulterants were analyzed. Zanthoxmli Pericarpium was i-dentified through the Species identification System for Traditional Chinese Medicine and neighbor-joining (NJ) phylogenetic trees. RESULTS: The lengths of ITS/ITS2 sequence of Zanthoxmli Pericarpium were 618-620 bp and 224 bp. The intra-specific genetic distances were smaller than inter-specific ones in ITS/ITS2 regions of Z. schinifolium and Z. bungeanum. Zanthoxmli Pericarpium and its adulterants could be easily differentiated according to the nearest distance, the Species Identification System for Traditional Chinese Medicine and the NJ trees methods. CONCLUSION: ITS/ITS2 regions as DNA barcodes can stably and accurately distinguish Zanthoxmli Pericarpium and its adulterants.
4.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
5.Role of asymmetric dimethylarginine in acute lung injury induced by cerebral ischemia/reperfusion injury in rats.
Yun-hu WU ; Xuan ZHANG ; Dian-hua WANG
Journal of Southern Medical University 2011;31(8):1289-1294
OBJECTIVETo determine the role of asymmetric dimethylarginine (ADMA) in acute lung injury induced by cerebral ischemia/reperfusion (I/R) injury in rats.
METHODSAdult male SD rats were randomly divided into 4 groups, namely the sham-operated group (S), cerebral I/R model group, ADMA+I/R group, and dimethylarginine dimethylaminohydrolase (DDAH)+I/R group. In the latter 3 groups, acute lung injury was induced by left middle cerebral artery occlusion for 120 min. After a 24-h reperfusion, the rats were sacrificed and the activities of nitric oxide synthase (NOS) and contents of nitric oxide (NO) were measured using reductase and colorimetric assay. The mRNA and protein expressions of protein kinase C (PKC) and myosin light chain kinase (MLCK) in the lung tissues were detected with RT-PCR and Western blotting, respectively. The contents of ADMA in the bronchoalveolar lavage fluid (BALF) and blood flowing into and out of the lungs were measured by ELISA.
RESULTSCerebral I/R injury caused significantly elevated ADMA levels in the BALF and blood flowing into the lungs, and obviously lowered the NO concentration and NOS activity in the lung tissues (P<0.05). Following cerebral I/R injury, MLCK and PKC mRNA and protein expressions were significantly upregualted in the lung tissues (P<0.05). Exogenous DDAH obviously decreased the levels of ADMA in the BALF and blood flowing into the lungs, increased NO concentration and NOS activity, and down-regulated MLCK and PKC mRNA and protein expressions in lung tissues of rats with cerebral I/R injury (P<0.05).
CONCLUSIONADMA contributes to the development of acute lung injury following cerebral I/R injury in rats by upregulating MLCK and PKC expression. ADMA may serve as a novel therapeutic biomarker and a potential therapeutic target for acute lung injury induced by cerebral I/R injury.
Acute Lung Injury ; etiology ; physiopathology ; Animals ; Arginine ; analogs & derivatives ; metabolism ; pharmacology ; Brain Ischemia ; complications ; Male ; Myosin-Light-Chain Kinase ; genetics ; metabolism ; Nitric Oxide Synthase ; antagonists & inhibitors ; Protein Kinase C ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; physiopathology ; Up-Regulation ; drug effects
6.Study on effect of pH on rectum permeability of active ingredients in Reduning suppositories in vitro.
Ming YAN ; Ying-chun WEI ; Jin MENG ; Dian-hong XU ; Yun WU ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(8):1493-1496
OBJECTIVETo investigate the effect of different pH on rectum permeability of chlorogenic acid and geniposide.
METHODFour kinds of Reduning suppositories of different pH were separated and put into the rectum to study the suppositories in vitro and the content of chlorogenic acid and geniposide samples was determined by HPLC to calculate the permeation in 24 hours.
RESULTWith increase of pH within 2.5-7.4, the steady state flux of chlorogenic acid was increased, but the steady state flux of geniposidesamples was steady.
CONCLUSIONAdjusted the pH can increase the rectum permeability of active ingredients in Reduning auppositories.
Animals ; Chlorogenic Acid ; pharmacokinetics ; Drugs, Chinese Herbal ; pharmacokinetics ; Hydrogen-Ion Concentration ; Iridoids ; pharmacokinetics ; Male ; Permeability ; Rats ; Rats, Sprague-Dawley ; Rectum ; metabolism ; Suppositories ; pharmacokinetics
7.Observation on effect of danxiaoling in supplementary treatment of intrahepatic cholestasis in pregnancy.
Dian-yun SHI ; Hua CHEN ; Mei XIAO
Chinese Journal of Integrated Traditional and Western Medicine 2002;22(2):116-118
OBJECTIVETo evaluate the curative effect of Danxiaoling Pill (DXLP), a Chinese herbal preparation, in treating intrahepatic cholestasis in pregnancy (ICP).
METHODSFifty-eight cases of ICP were divided randomly into two groups and treated by DXLP and Composite Yiganling as control respectively with the other identical conventional treatment. The changes of clinical symptoms, related laboratory parameters after treatment and the condition of labor were observed.
RESULTSThe total effective rate in both groups was 100%, but the markedly effective rate in the DXLP treated group was higher than that in control group (P < 0.01). Levels of blood cholyglycine acid (CGA), alanine transaminase (ALT), aspartate transaminase (AST), total bilirubin, total cholesterol and low density lipoprotein were all decreased in both groups after treatment, but DXL showed a better efficacy in decreasing CGA, ALT and AST than that of Yiganling. Moreover, the amniotic fluid meconium contaminated rate, premature delivery occurrence in the DXLP group were lower than those in the control group, while the weight of newborn baby was higher in the former than in the latter.
CONCLUSIONDXLP could effectively lower the serum bile acid and improve liver function in treating ICP.
Adult ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Cholestasis, Intrahepatic ; blood ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glycocholic Acid ; blood ; Humans ; Phytotherapy ; Pregnancy ; Pregnancy Complications ; blood ; drug therapy
8.Identification of peucedani radix, peucedani decursivi radix and its adulterants using ITS2 sequence.
Dian-Yun HOU ; Jing-Yuan SONG ; Pei YANG ; Hong ZHOU ; Tian-Yi XIN ; Hui YAO
China Journal of Chinese Materia Medica 2014;39(21):4186-4190
In order to identify Peucedani Radix, Peucedani Decursivi Radix and their adulterants, the internal transcribed spacer 2 (ITS2) regions of Peucedani Radix, Peucedani Decursivi Radix and their adulterants were amplified and bidirectionally sequenced based on the Principles for Molecular Identification of Traditional Chinese Materia Medica Using DNA Barcoding, which has been promulgated by Chinese Pharmacopoeia Commission. Sequences were analyzed and assembled by Codon Code Aligner V3. 7.1. The relevant data were analyzed by MEGA 5. 0. Species identification analyses were performed by using the nearest distance methods and neighbor-joining (NJ) methods. The result showed that the ITS2 sequence lengths of Peucedani Radix were 229-230 bp and the average intra-specific genetic distances were 0.005. The ITS2 sequence lengths of Peucedani Decursivi Radix were 227 bp and the sequences contained no variation site. The average inter-specific K2P genetic distance of Peucedani Radix, Peucedani Decursivi Radix and their adulterants species were 0.044 and 0.065 respectively. The minimum inter-specific divergence is larger than the maximum intra-specific divergence of Peucedani Decursivi Radix. The nearest distance methods and NJ trees results indicated that Peucedani Radix, Peucedani Decursivi Radix and their adulterants species could be identification clearly. The ITS2 regions can stably and accurately distinguish Peucedani Radix, Peucedani Decursivi Radix and their adulterants.
Apiaceae
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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, Ribosomal Spacer
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Drug Contamination
9.Survey of adult carotid atherosclerosis in water-related endemic fluorosis areas in Heilongjiang province in 2008
Jia-yong, LIU ; Hui, LIU ; Wei, DONG ; Ben, GAO ; Yun-qi, LIU ; Dian-jun, SUN
Chinese Journal of Endemiology 2010;29(6):634-636
Objective To investigate the disease situation of adult carotid atherosclerosis in water-related endemic fluorosis areas in Heilongjiang province in 2008 so as to explore the relationship between water-related endemic fluorosis and incidence of carotid atherosclerosis disease. Methods A total of 266 participants over the age of 40 from four villages in Zhaozhou county with water fluoride ≥ 1.0 mg/L in a fluorosis area and 283 residents over the age of 40 from four villages in Tailai county with water fluoride < 1.0 mg/L in a nonfluorosis area were investigated. A portable-type B mode color ultrasound was used to examine the left carotid artery of all participants.The carotid atherosclerosis was diagnosed and graded through the ultrasonograms. Results The prevalence rates of carotid atherosclerosis in the 4 fluorosis villages(Xinfeng, Taipingshan, Baochan and Houzheng villages) were 47.3% (35/74), 63.5%(40/63), 73.3%(33/45) and 60.7%(51/84), respectively, and in the 4 nonfluorosis villages(Hala,Qianxing, Sanjia and Ailin villages) were 32.7% (17/52), 32.9% (24/73), 39.2% (31/79) and 30.4% (24/79),respectively. The prevalence rates of carotid atherosclerosis in every villages standardized by age were 47.3% ,63.5%,73.3% ,60.7% and 34.7% ,36.3% ,43.0% ,41.3%, respectively. Statistic method used was Wilcoxon two sample test and the differences were significant(T = 26, P < 0.05 ). Total carotid atherosclerosis positive rate standardized by age in the 4 fluorosis villages was significantly higher than that in the 4 nonfluorosis villages[57.5%(153/266) ,37.8% (107/283), x2 = 21.36, P < 0.01 ]. After standardized by age, the severity of carotid atherosclerosis was significantly different between fluorosis villages and nonfluorosis villages (x2 = 36.15, P < 0.01 ). Conclusion The prevalence rate of carotid atherosclerosis in endemic fluorosis area is higher than that in nonfluorosis area.
10.Integrin and Foot-and-mouth Disease Virus Infection
Jun-Zheng DU ; Hui-Yun CHANG ; Shan-Dian GAO ; Xue-Peng CAI ;
Microbiology 1992;0(05):-
Integrins are a family of cell surface glycoproteins that contribute to a variety of biological functions, including cell growth, migration, proliferation and morphology. In addition, integrins also play the important roles in pathological process. Several viruses have been showed to use integrins as receptors or co-receptors to infect host cells.This article mainly reviews the progress on integrins and their roles in FMDV infection.