1.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
2.A new flavone C-glycoside from leaves of Lophatherum gracile.
Hui-Nan ZHAO ; Mei CHEN ; Chun-Lin FAN ; Ying WANG ; Yao-Lan LI ; Wen-Cai YE
China Journal of Chinese Materia Medica 2014;39(2):247-249
Several kinds of column chromatography method were used to investigate the chemical constituents of the leaves of Lophatherum gracile. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. A new flavone C-glycoside was isolated and its structure was identified as 3'-methoxyl-luteolin 6-C-beta-D-galactopyranosiduronic acid (1 --> 2) -alpha-L-arabinopyranoside (1).
Antiviral Agents
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chemistry
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isolation & purification
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Flavones
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chemistry
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Glycosides
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chemistry
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isolation & purification
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Hydrolysis
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Plant Leaves
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chemistry
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Poaceae
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chemistry
3.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
4.Research achievements on structures and activities of polysaccharides from Panax ginseng.
Shan-shan LI ; Yin-ping JIN ; Chun-lin YAO ; Ying-ping WANG
China Journal of Chinese Materia Medica 2014;39(24):4709-4715
Panax ginseng C. A. Meyer (P. ginseng) has been used as traditional medicine in Asian countries for more than 2,000 years. P. ginseng contains many active components such as ginsenosides, peptides, essential oil and polysaccharides, among which, P. ginseng polysaccharides were reported to have immunomodulating, anti-cancer, anti-adhesive and antioxidant activities. For better understanding of the structures and biological activities of all the ginseng polysaccharides, here the recent research achievements were reviewed. This review would be helpful for the relevant researchers to get useful information.
Antineoplastic Agents
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analysis
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isolation & purification
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pharmacology
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Antioxidants
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analysis
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isolation & purification
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pharmacology
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Humans
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Immunologic Factors
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analysis
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isolation & purification
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pharmacology
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Medicine, Traditional
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Panax
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chemistry
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Plant Roots
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chemistry
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Plants, Medicinal
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Polysaccharides
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analysis
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isolation & purification
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pharmacology
5.Preparation and pharmacokinetic evaluation of long-acting injectable oily suspensions for ophiopogonis radix polysaccharide MDG-1.
Xiao-Li SHI ; Chun-Xia YAO ; Xiao LIN ; Lan SHEN ; Yi FENG
China Journal of Chinese Materia Medica 2014;39(13):2489-2494
OBJECTIVETo evaluate in vivo pharmacokinetics of Ophiopogonis Radix polysaccharide MDG-1 oily suspension injection prepared with different prescriptions in rats, and explore the feasibility of the long-acting drug delivery of MDG-1 Injection by using the oily suspension drug release system.
METHODMDG-1 microparticles were prepared by the anti-solvent precipitation method. Their size and size distribution were characterized. Castor oil with a high viscosity or aluminum stearate were added into soybean oil with a low viscosity, in order to prepare oily media with different viscosities, detect their rheological properties and screen out superior prescriptions for in vivo evaluation.
RESULTThe average size of microparticles was 21.81 microm, and the span between them was 2.63. The in vivo evaluation was conducted for prescriptions of mixed oil (soybean oil/castor oil, 2: 3) and soybean oils gelled by 2% and 4% aluminium stearate. Among them, the prescription of soybean gelled by 4% aluminium stearate could significantly reduce C(max) and prolong the apparent t1/2, with the MDG-1 release time of several days.
CONCLUSIONIt is feasible to achieve the long-acting MDG-1 drug delivery by using oily media with a high viscosity.
Animals ; Chemistry, Pharmaceutical ; methods ; Drug Delivery Systems ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Male ; Ophiopogon ; chemistry ; Plant Oils ; chemistry ; Polysaccharides ; administration & dosage ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Viscosity
6.Integrated DNA barcoding database for identifying Chinese animal medicine.
Lin-Chun SHI ; Hui YAO ; Li-Fang XIE ; Ying-Jie ZHU ; Jing-Yuan SONG ; Hui ZHANG ; Shi-Lin CHEN
China Journal of Chinese Materia Medica 2014;39(12):2155-2159
In order to construct an integrated DNA barcoding database for identifying Chinese animal medicine, the authors and their cooperators have completed a lot of researches for identifying Chinese animal medicines using DNA barcoding technology. Sequences from GenBank have been analyzed simultaneously. Three different methods, BLAST, barcoding gap and Tree building, have been used to confirm the reliabilities of barcode records in the database. The integrated DNA barcoding database for identifying Chinese animal medicine has been constructed using three different parts: specimen, sequence and literature information. This database contained about 800 animal medicines and the adulterants and closely related species. Unknown specimens can be identified by pasting their sequence record into the window on the ID page of species identification system for traditional Chinese medicine (www. tcmbarcode. cn). The integrated DNA barcoding database for identifying Chinese animal medicine is significantly important for animal species identification, rare and endangered species conservation and sustainable utilization of animal resources.
Animals
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DNA Barcoding, Taxonomic
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methods
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Databases, Nucleic Acid
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Eukaryota
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classification
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genetics
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Medicine, Chinese Traditional
7.Efficacy and safety of diffirent hormonal therapy regiments in advanced prostate cancer patients
Hai HUANG ; Kewei XU ; Jian HUANG ; Wen DONG ; Chun JIANG ; Tianxin LIN ; Zhenghui GUO ; Yousheng YAO ; Wenlian XIE ; Jinli HAN
Chinese Journal of Urology 2010;31(1):45-48
Objective To compare the efficacy,toxicitis and side-effects of Casodex and Flutamide in the hormonal therapy of advanced prostate cancer patients.Methods One hundred and thirty-six advanced prostate cancer patients were treated with with hormonal therapy.The patients were divided into 3 groups,of which 52 patients (group A) used LHRHa and Casodex as intermittent hormonal therapy;60 patients(group B) used LHRHa and Flutamide as intermittent hormonal therapy;24 patients(group C) were treated with surgical castration only.The difference of clinical symptoms,serum PSA,disease progression risk,survival rate,toxicitis and side-effects of 3 groups were compared.Results The relief rates of group A and B were 80.8% (42/52)and 81.7% (49/60) respectively,higher than 70.8% (17/24) of group C.The mean serum PSA of group A and B decreased from 133.3 ng/ml(17.9-982.8 ng/ml) to 15.8 ng/ml(0.02-28.9 ng/ml),142.6 ng/ml (20.2-1001.0 ng/ml)to 16.1 ng/ml(0.07-53.8 ng/ml),respectively,both better than that of group C,which decreased from 142.3 ng/ml (27.1-988.0 ng/ml) to 27.6 ng/ml(6.0-62.1 ng/ml).The mean chemical recurrence rates of group A and B were 34.7% (18/52) and 36.7% (22/60),respectively,lower than 58.3% (14/24) of group C.The mean chemical recurrence time of group A and B was 22(5-52)months and 22(6-65)months,respectively,longer than 11(5-54)months of group C.The mortality rates of group A and B were 26.9% (14/52) and 31.7% (19/60),respectively,lower than 66.7%(16/24) of group C.88.5% (46/52)of group A were treated continuously,while group B had 66.7% (40/60).The side-effects rate of group A was lower than group B.Conclusions Both Cadosex and Flutamide are effective for prostate cancer,and decrease the disease progression risk.Casodex is more effective and safer as for the treatment of prostate cancer compared to Flutamide.
8.Nephrectomy and removal of inferior vena cava tumor thrombus under profound hypothermia and arrested circulation
Yousheng YAO ; Jian HUANG ; Hai HUANG ; Yanqi YANG ; Shuling PENG ; Kewei XU ; Zhenghui GUO ; Tianxin LIN ; Chun JIANG ; Jinli HAN
Chinese Journal of Urology 2008;29(5):300-302
Objective To study the feasibility and safety of performing nephrectomy together with the removal of complicated inferior vena cava tumor thrombus under profound hypothermia and arrested circulation. Methods After made the median thoraco-abdominal incision, the exploration of the abdominal organs was done. The right kidney, inferior vena cava and renal pedicle were well exposed then. After the whole body heparinization, cannulas were put into ascending aorta, superior vena cava, aortic root and right superior pulmonary vein. The body temperature was reduced to 20℃ with cardiopulmonary bypass unit and the extracorporeal circulation was stopped then. Cut open the inferior vena cava at vena renalis dextra ingress and the F16 urinary catheter was inserted into atrum dextra through inferior vena cava and inflated. The tumor thrombus was pulled out and the right kidney was removed. The inferior vena cava incision was sutured to close and the extracorporeal circulation was resumed and patient was re-warmed.Results The operation time was 330 min and the extracorporeal circulation time was 90 min, while the profound hypothermia with circulatory arrest time was 20 min. The estimated blood loss during operation was 400 ml and 6 unit red cells and 600 ml blood plasm were transfused. The patient was awaked 2.5 h after the operation, food intake resumed 4 days after operation and the patient was discharged on day 10 post-operatively. After 6 months'follow-up, there were no local recurrence and metastasis occurred. Conclusion The technique of profound hypothermia and circulation arrest could improve the safety and efficacy in the treatment of renal cell carcinoma with suprahepatic (level Ⅲ) caval tumor thrombus.
9.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
10.Crypotanshione reduces the expression of metadherin in DU145 prostate cancer cells.
Yi YAO ; Hui-zhang LI ; Ben-jiang QIAN ; Chang-ming LIU ; Jia-bin ZHANG ; Miao-chun LIN
National Journal of Andrology 2015;21(9):782-787
OBJECTIVETo investigate the effects of crypotanshinone (CPT) on the proliferation and apoptosis of DU145 prostate cancer cells as well as on the metadherin expression and the downstream PI3K/AKT signaling pathway in the DU145 cells.
METHODSWe treated DU145 prostate cancer cells with different concentrations of CPT for 24, 48, and 72 hours followed by evaluation of the proliferation and apoptosis of the cells by MTT assay and TUNEL, respectively. We determined the expressions of metadherin protein and mRNA in the DU145 cells by Western blot and RT-PCR respectively at different time points after CPT treatment. We also detected the expressions of the proteins metadherin, AKT, p-AKT, and Bcl-2 in the CPT-treated DU145 cells at 48 hours.
RESULTSCPT significantly inhibited the proliferation of the DU145 cells in a dose- and time-dependent manner (P < 0.05). After treatment with 10 µmol/L CPT for 24, 48, and 72 hours, the apoptosis rates of the DU145 cells were (29.42 ± 4.51), (55.07 ± 5.67) and (70.84 ± 4.66)%, respectively, significantly higher than (3.1 ± 2.48)% in the control group (P < 0.05). The expression of metadherin was remarkably downregulated at the transcription and translation levels (P < 0.05) and the expressions of the AKT signaling pathway and the Bcl-2 protein were markedly inhibited in the DU145 cells after treated with 10 µmol/L CPT for 48 hours (P < 0.05).
CONCLUSIONCPT can inhibit the proliferation and induce the apoptosis of DU145 prostate cancer cells, which may be associated with its suppression of the downstream PI3K/AKT signaling pathway by reducing the expression of metadherin in the DU145 cells.
Apoptosis ; drug effects ; Cell Adhesion Molecules ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Abietane ; pharmacology ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Humans ; In Situ Nick-End Labeling ; Male ; Neoplasm Proteins ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; drug effects ; Time Factors