1.Establishment of adventitious root culture system and scale-up fermentation of Tripterygium wilfordii.
Yan LI ; Lei CUI ; Yu-Qi YANG ; Lei ZHAO ; Jia-Min LEI ; Xing ZHANG
China Journal of Chinese Materia Medica 2015;40(1):53-58
Using MS as basic medium, supplemented with 1.0 mg · L(-1) IBA, the adventitious roots of Tripterygium wilfordii were induced, and the good adventitious root culture system was established by leaves or callus induced by leaves as explants. The adventitious roots were also induced with 2.0-4.0 mg · L(-1) NAA and the good adventitious root culture system established by using suspension cells from callus as materials to induce adventitious root. The content of triptolide of three adventitious roots culture system were exceeded in the natural root bark. The content of triptolide of AR3 adventitious roots was the highest about 5.3 times as that in the natural root bark. By using 5 L stirred fermentor during pilot enlarge cultivation, compared with 250 mL flask cultivation, the adventitious roots increment and secondary metabolites content per liter medium showed no significant difference. The accomplishment of this analysis laid a foundation by tissue culture production of the secondary metabolites of T. wilfordii.
Culture Media
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chemistry
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metabolism
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Fermentation
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Plant Growth Regulators
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analysis
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metabolism
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Plant Roots
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growth & development
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metabolism
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Tissue Culture Techniques
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instrumentation
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methods
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Tripterygium
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growth & development
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metabolism
2.Effects of amino acid on growth and secondary metabolites contents of adventitious roots of Tripterygium wilfordii.
Peng-Fei WEN ; Jia-Min LEI ; Qun LI ; Lei CUI ; Yan LI ; Xing ZHANG
China Journal of Chinese Materia Medica 2014;39(12):2267-2274
The adventitious root of Tripterygium wilfordii was used as experiment material to study effects of various concentration of aspartic acid, isoleucine, cysteine and arginine in MS medium on the growth and triptolide, wilforgine, wilforine contents of the adventitious roots. The results showed that compared with the control, supplemented with 0.25 mmol x L(-1) aspartic acid at 3rd week, the growth of the adventitious roots only accounted for 80%, but the content of triptolide of the adventitious roots and the medium was 1.36, 1.30 times, the content of wilforgine was 1.16, 1.37 times, the content of wilforine was 1.22, 1.63 times, respectively. At 3rd week 0.05 mmol x L(-1) isoleucine, the growth of adventitious roots was 97.3%, wilforgine of adventitious roots and medium 1.02, 1.27 times, wilforine 1.36 times and 1.15 times. At 1st week 0.25 mmol x L(-1) cysteine, the growth of the adventitious roots comprised 77.5% of the control, while content of triptolide of adventitious roots reached 1.87 times. At 2nd week 1.00 mmol x L(-1) cysteine, the growth of adventitious roots was 44.6% of the control, the content of wilforine in medium was 2.97 times. At 3rd week 0.50 mmol x L(-1) arginine, the growth of adventitious roots was 124.2%, the content of wilforgine and wilforine was 1.3, 1.4 times, respectively.
Amino Acids
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analysis
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metabolism
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Diterpenes
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analysis
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Epoxy Compounds
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analysis
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Lactones
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analysis
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metabolism
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Phenanthrenes
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analysis
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Plant Roots
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chemistry
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growth & development
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metabolism
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Pyridines
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analysis
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metabolism
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Secondary Metabolism
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Tripterygium
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chemistry
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growth & development
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metabolism
3.Safety Evaluation for Interventional Radiotherapy with X-ray Angiography
Yani BAI ; Shu QIAN ; Zifu HE ; Min JIA ; Xiao FAN ; Lei JIN
Chinese Medical Equipment Journal 2003;0(10):-
Objective To evaluate the safety of interventional radiotherapy with X-ray angiography. Methods To compare X-ray radiation levels with and without radioprotect with X-ray protective monitor by simulating operators' body's parts. Results The difference is significant. Conclusion Interventional radiotherapy with angiography with effective protect is safe.
4.Expression of mCD_(14) on Alveolar Macrophage of Neonates with Respiratory Distress Syndrome Treated by Me-chanicalventilation
li-min, KANG ; lei, CAO ; cui-qing, LIU ; xi-qun, JIA ; qiu-jin, LIANG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To investigate the mechanism of mCD14 expression on AM in the pathogenes of neonatal respiratory distress syndromes( NRDS). Methods The expression of mCD14 on AM was analyzed with flow cytometry. Enzyme - linked immunosorbent assay was performed for detecting the concentration of IL- 1? and IL-8.Results The percentage of mCD14 positive AM in experimental group [(54.772 ?17 .341)%] was higher than that in control group [(14.023? 10. 713)% ](t= -7.739 P
5.Expression and characterization of a novel halohydrin dehalogenase from Tistrella mobilis KA081020-065.
Lei WANG ; Jing YUAN ; Peiyuan YAO ; Lihua CHENG ; Meixian XIE ; Rongrong JIA ; Huijin FENG ; Min WANG ; Qiaqing WU ; Dunming ZHU
Chinese Journal of Biotechnology 2015;31(5):659-669
Halohydrin dehalogenase is of great significance for biodegradation of the chlorinated pollutants, and also serves as an important biocatalyst in the synthesis of chiral pharmaceutical intermediates. A putative halohydrin dehalogenase (HheTM) gene from Tistrella mobilis KA081020-065 was cloned and over-expressed in Escherichia coli BL21 (DE3). The recombinant enzyme was purified by Ni-NTA column and characterized. Gel filtration and SDS-PAGE analysis showed that the native form of HheTM was a tetramer. It exhibited the highest activity at 50 degrees C. The nature and pH of the buffer had a great effect on its activity. The enzyme maintained high stability under the alkaline conditions and below 30 degrees C. HheTM catalyzed the transformation of ethyl(S)-4-chloro-3-hydroxybutyrate in the presence of cyanide, to give ethyl (R)-4-cyano-3-hydroxybutyrate, a key intermediate for the synthesis of atorvastatin.
3-Hydroxybutyric Acid
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chemistry
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Bacterial Proteins
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genetics
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metabolism
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Cloning, Molecular
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Escherichia coli
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Hydrolases
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genetics
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metabolism
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Hydroxybutyrates
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chemistry
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Recombinant Proteins
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genetics
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metabolism
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Rhodospirillaceae
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enzymology
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genetics
7.In vitro effect of total flavones of Fructus Chorspondiatis on expression of collagen type I and type III mRNA and protein of cultured rat cardiac fibroblasts.
Jun-Ping BAO ; Ming JIN ; Yu-Min YANG ; Xiao-Hui GAO ; Liang SHU ; Hui-Hui XING ; Lei JIA
Acta Pharmaceutica Sinica 2014;49(1):136-141
This study aims to investigate the effect of total flavones of Fructus Chorspondiatis (TFFC) on the mRNA and protein expression of collagen type I and III of rat cardiac fibroblasts (CFs) induced by angiotensin II (Ang II), and explore its anti-myocardial fibrosis molecular mechanism. Neonatal rat CFs were prepared from Sprague-Dawley rats (1-3 d after birth). The expression of collagen type I and III mRNA and protein were measured by RT-PCR and Western blotting, respectively. The study showed that stimulation of neonatal rat CFs with 100 nmol.L-1 of Ang II for 72 h resulted in a significant increase of the expression of collagen type I and III mRNA and protein. The changes on the expression level were blocked by TFFC. The results demonstrated that TFFC can inhibit myocardial fibrosis induced by Ang II in rats, which is probably associated with the collagen type I and III mRNA and protein levels up-regulated by Ang II, and TFFC was shown to decrease the expression levels of collagen type I and III mRNA and protein.
Anacardiaceae
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chemistry
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Angiotensin II
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pharmacology
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Animals
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Animals, Newborn
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Cells, Cultured
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Collagen Type I
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genetics
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metabolism
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Collagen Type III
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genetics
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metabolism
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Dose-Response Relationship, Drug
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Drugs, Chinese Herbal
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administration & dosage
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isolation & purification
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pharmacology
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Fibroblasts
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cytology
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metabolism
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Flavones
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administration & dosage
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isolation & purification
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pharmacology
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Fruit
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chemistry
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Myocardium
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cytology
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metabolism
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Plants, Medicinal
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chemistry
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RNA, Messenger
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metabolism
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Rats
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Rats, Sprague-Dawley
8.The mechanism of toxicity enhancement with the combination of Flos Genkwa and Radix et Rhizoma Glycyrrhizae based on the solubilization of glycyrrhizic acid
Yu-qin YANG ; Fei-fei LI ; Shan CHEN ; Zhi-jia WANG ; Peng-long WANG ; Hai-min LEI
Acta Pharmaceutica Sinica 2021;56(9):2561-2566
In order to study the contraindications of the compatibility of Flos Genkwa-Radix et Rhizoma Glycyrrhizae, in this study, the solubilizing and poisoning essence were explored. In this experiment, chromatographic assay, field emission scanning electron microscopy, MTT cytotoxicity evaluation, and other methods were used to study the main chemical components, morphology and toxicity of the ethyl acetate part of Flos Genkwa and its co-decoction with glycyrrhizic acid, in order to clarify Flos Genkwa-Radix et Rhizoma Glycyrrhizae incompatibility provides a new idea for the research on incompatibility of Flos Genkwa-Radix et Rhizoma Glycyrrhizae. The results showed that after co-decoction of the ethyl acetate part of Flos Genkwa with glycyrrhizic acid, high performance liquid chromatography (HPLC) detected the dissolution of the toxic component yuanhuacine of 54.8%, while yuanhuacine chromatographic peak was not detected in the Flos Genkwa ethyl acetate part of the single decoction. The increase of co-decoction dissolution rate was observed by scanning electron microscopy, and it was found that glycyrrhizic acid uniformly dispersed the fat-soluble components of Flos Genkwa into nano-scale particles, which improved the solubility and stability in the solution. Furthermore, the results of cytotoxicity evaluation showed that the survival rate of cells decreased after co-decoction, 4',6-diamidino-2-phenylindole (DAPI) staining also gave the same results. In summary, the co-decoction of the ethyl acetate part of Flos Genkwa with glycyrrhizic acid promotes the dissolution of the toxic component yuanhuacine, and makes the part form uniformly distributed nanoparticles, which is conducive to the absorption of the ingredient and increases the toxicity.
9.Based on weak bond chemistry, the interaction mechanism between glycyrrhiza protein and berberine in water decocting process of Rhizoma Coptidis and Liquorice was investigated
Wen LI ; Zhi-jia WANG ; Xiao-jing LIU ; Na-na HAN ; Tong LI ; Hai-min LEI ; Peng-long WANG
Acta Pharmaceutica Sinica 2021;56(8):2119-2126
The purpose of this study was to explore the interaction mechanism between glycyrrhiza protein and berberine in the decocting process of
10.Effects of enrichment with toys on growing development and reproductive performance of mice
Lei LIANG ; Longxiang XU ; Jiakuan LIU ; Li CHEN ; Chunmei JIA ; Jinwei YOU ; Min DONG ; Wenjuan HU ; Shifeng YUN
Chinese Journal of Comparative Medicine 2014;(9):48-52
Objective To study the effects of toys for laboratory animals on reproductive performance and growing development of experimental mice.Methods ICR mice were fed with toys, the informations of reproductive performance and growing development were recorded.Results All the data of reproductive performance of the test group were higher than the control group except the number of newborn mice, and showed significant difference ( P <0.05 ) .The data of growing development of the test group were higher than the control group too, but showed no significant difference ( P >0.05).Conclusion Toys for laboratory animals have good effects on reproductive performance and growing development of mice, and suggested to be used into the process of experimental mice raising.