Limonoids from seeds of A. Juss. and their cytotoxic activity

Four new limonoid-type nortriterpenoids, 1-detigloyl-1--methacryloylsalannin (), 28-deoxo-2,3-dihydronimbolide (), 12-acetoxy-3--acetyl-7--tigloylvilasinin () and 12-acetoxy-3--acetyl-7--methacryloylvilasinin (), along with five known ones, were isolated from seeds of A. Juss. Their structures were elucidated by various spectroscopic methods, including UV, IR, MS, NMR, X-ray crystallography, quantum chemical calculation, as well as by comparison of their spectroscopic data with those reported. In the cytotoxic assay, showed inhibitory activity against human breast cancer MDA-MB-231 cell line with IC value of 7.68±1.74 μmol/L, and inhibited growth of human cervical cancer Hela cell line, melanoma A375 cell line and promyelocytic leukemia HL-60 cell line, with IC 12.00±2.08, 17.44±2.11, and 13.95±5.74 μmol/L, respectively.

Study progress on antitumor activities of artemisinin and its derivatives / 中国生化药物杂志

The antitumor activities of artemisinin and its derivatives has become a hot topic in recent years.Artemisinin and its derivatives have been reported to exert antitumor effects mainly by blocking the cell cycle, promoting tumor cell apoptosis, inhibiting angiogenesis and so on.Also, their influence on signaling pathways has attracted considerable interest of many scholars.Currently, in order to discover some compounds with better therapeutic potentials, many studies dedicated to the synthesis and development of new artemisinin derivatives.In addition, the effects of artemisinin and its derivatives on multi-drug resistantace in tumor cells also become a trend of the future research.In this paper, we review the research advances in the antitumor activities of artemisinin and its derivatives during the past five years to provide a reference for future study .

Connexin: a new perspective of tumor therapy / 中国生化药物杂志

Connexin (Cx) is the main component of gap junction channels, which is essential for intercellular communication between adjacent cells.Cx plays an important role in regulation of the growth, differentiation and apoptosis of cells and is closely related to tumor formation and development.The expression of Cx appears to be different in different stages of tumor development, and the regulatory mechanism of Cx is also involved in many aspects.In this paper, recent advances in the studies on Cx are reviewed, including the relationship between Cx and development of tumor, the basic mechanism of Cx, as well as its applications in antitumor-drug screening and tumor therapy.

Impact of 2, 3-dihydrocyclopentacoumarins to growth of transgenic hairy roots of Polygonum multiflorum and accumulation of stilbene glucosides / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the influence of 2,3-dihydrocyclopenta coumarins to the growth of transgenic hairy root of Polygonum multiflorum and the accumulation of active ingredient stilbene glucoside in the culture.</p><p><b>METHOD</b>In different culture time, 2,3-dihydrocyclopenta coumarin compounds were administrated to the hairy roots. The yield of stilbene glucoside and the growth of the hairy roots were determined after some period of co-culture. Through the investigation of the different concentrations of the 2,3-dihydrocyclopentacoumarin, and sub-culture times, the optimal experimental conditions were established.</p><p><b>RESULT</b>The optimal administration time of the 2,3-dihydrocyclopentacoumarin was on the fourth day, the optimal concentration of 2,3-dihydrocyclopentacoumarin was 0.025 g x L(-1). Under this condition, the growth of the hairy root greatly increased. Comparing with control group, the accumulation of stilbene glucoside was increased about four times.</p><p><b>CONCLUSION</b>2, 3-dihydrocyclopentacoumarins could promote the hairy hair root growth and stimulate the stilbene glucoside accumulation.</p>

Biotransformation of daphnetin by suspension transgenic hairy roots of Polygonum multiflorum / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the biotransformation of daphnetin by suspension transgenic hairy root of Polygonum multiflorum and provide a biotechnological method for large-scale production of the daphnetin-8-O-beta-D-glucoside using this new culture system.</p><p><b>METHOD</b>Daphnetin was added into the media of suspension to culture 36 h. The biotransformation product was detected with TLC and HPLC, and isolated by various chromatographic methods. The influence of co-cultured time on conversion ratio, content of degradation product and the reason for the degradation of product II were investigated using HPLC.</p><p><b>RESULT</b>One biotransformation product, daphnetin-8-O-beta-D-glucoside, was obtained, the optimal co-cultured time in suspension hairy root of P. multiflorum was 36 h with the highest biotransformation molar ratio of 32.11%, the sucrose medium (sucrose-only) can increase the biotransformation molar ratio to 72.44%. The result demonstrated that the degradation products of the product II was induced by the MS medium.</p><p><b>CONCLUSION</b>The potential application of suspension transgenic hairy root of P. multflorum in the sucrose-only medium on generating daphnetin-8-Obeta-D- glucoside could be prospective.</p>

Studies on anti-EV71 virus activity of traditional Chinese medicine and its clinical application in treatment of HFMD / 中国中药杂志

Hand-foot-and-mouth disease is caused by intestinal virus infection. The viruses coxsackie A16 (CA16) and enterovirus 71 (EV71) are the main pathogens. Between them, the virus EV71 is more dangerous and easier to cause serious complications, which leads to death or disability. Currently, there are no effective antiviral drugs to treat EV71 infection. Therefore, developing an effective drug against EV71 virus activity is significant. It has a huge potency of screening the anti-EV71 components and developing the new drugs from the abundant traditional Chinese medicines (TCMs). Meanwhile, since hand-foot-and-mouth disease spread in Shanghai in 1981, a growing number of reports on TCMs treatment in clinic have been published. In addition, most of treatments with various ways are effective, which play a positive role on improving clinic treatment and controlling diseases. Moreover, special clinic advantages and features of TCMs were obviously shown.

Preliminary study on promoting effects of endophytic fungi to growth of Rehmannia glutinosa / 中国中药杂志

In previous studies, four endophytic fungi were isolated from different swollen roots of Rehmannia glutinosa. It's thought that Ceratobasidium sp. , one of the discovered endophytic fingi, was a major promoter for the growth of the roots. In this study, symbiotic experiments were performed to measure the effects of different endophytic fingi cultivated with R. glutinosa. The results showed that the R. glutinosa had significant increases in the size of roots and amount of chlorophyll cultivated with Ceratobasidium sp. And it was tested that indoleacetic acid secreted by Ceratobasidium sp. maybe the effective factor for the promotion of the growth.

Isolation and identification of endophytic fungi from different swollen root of Rehmannia glutinosa / 中国中药杂志

The swollen root of Rehmannia glutinosa is used as one kind of important Chinese traditional medicine. The root of R. glutinosa usually swelled in rotational cropping but not in continuous cropping. The rhizosphere microorganisms of R. glutinosa under different farming condition were thought related to that. In this study, the endophytic fungi in the root of R. glutinosa growing in various soil conditions were isolated for the study of the relationship between the microorganisms and the root enlargement of their host plants. The dominant endophytes, Verticillium spp., Fusarium oxysporum, F. redolens and Ceratobasidium spp. were identified by morphological observation and 18S rDNA and ITS sequence analysis. The preliminary investigation showed that the excessive growth of Verticillium and Fusarium genus fungi is unfavorable for the R. glutinosa root swelling, but Ceratobasidium fungi has no effects on the root enlargement.

Antioxidative and cytotoxic properties of diarylheptanoids isolated from Zingiber officinale / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the antioxidant and cytotoxic properties of five diarylheptanoids (1-5) isolated from the rhizomes of Zingiber officinale.</p><p><b>METHOD</b>Various models such as scavenging superoxide anions and 1,1-diphenyl-2- picrylhydrazyl (DPPH) radicals, inhibiting lipid peroxidation, as well as protecting of rat pheochromocytoma (PC12) cells induced by hydrogen peroxide (H2O2) were employed to assay the antioxidative effects of the diarylheptanoids. The cytotoxicities of compounds 1-5 were measured with MTT assays.</p><p><b>RESULT</b>The test compounds (1-5) showed promising DPPH inhibitory activities, and compound 5 exhibited the strongest DPPH scavenging activity with an IC50 value of (22.6+/-2.4) micromol x L(-1). Compounds 1, 3 and 4 showed potential anti-peroxidative effects with inhibitory rates of (66.3+/-15.4)%, (68.7+/-15.8)% and (72.2+/-10.6)%, respectively, at 100 microg x mL(-1). It could be observed that compounds 1, 3 and 4 demonstrated significant neuroprotective activities in a dose-dependent manner. Moreover, compound 3 exhibited certain cytotoxicities against human chronic myelogenous leukemia cells (K562) and its adriamycin-resistant cells (K562/ADR) with IC50 values of (34.9+/-0.6), (50.6+/-23.5) micromol x L(-1), respectively.</p><p><b>CONCLUSION</b>In vitro results demonstrated that five diarylheptanoids (1-5) isolated from the roots of Z. officinale were capable of scavenging radicals, inhibiting lipid peroxidation and protecting PC12 cells against the insult by H2O2. Additionally, compound 3 could inhibit the growth of K562 and K562/ADR cells.</p>

Biosynthesis and biotransformation of biological active constituents from natural medicines / 中草药

Bioactive constituent is the substantial basis of treatment for various diseases by Chinese materia medica (CMM) and also one of the most important substances for the modernization of CMM. Biosynthesis and biotransformation are considered to be one of the mild systems in bio-catalysis with optimal efficiency and selectivity, which play more potential role in the field, such as traditional Chinese medicine. To review and discuss some applications of biosynthesis and biotransformation in the research of CMM and natural medicines and their relationship of modernization of CMM as well. Moreover, a prospect is given exploringly.

Biotransformation of dihydroartemisinic acid by suspension culture cell of Artemisia annua / 中草药

Objective To investigate the biotransformation of dihydroartemisinc acid(Ⅰ) by cultured cells of Artemisia annua.Methods Dihydroartemisinic acid was added to the suspension cells of A.annua which had been pre-cultured for 14 d and co-cultured for another 2 d.The biotransfromed products were detected with TLC and HPLC,and isolated by various chromatographic methods.Results The chemical structures of biotransformed products were elucidated on the basis of spectroscopic data.3-?-Hydroxy-dihydroartemisinic acid(Ⅱa) and 3-?-hydroxy-dihydroartemisinic acid(Ⅱb) were obtained after 2 d administration of dihydroartemisinic acid to the suspension cells of A.annua.Ⅱa and Ⅱb yields could reach a maxium mole ratio of 2.6% and 15.7% after 1 d incubation,respectivly.Conclusion It is the first time for the biotransformation of dihydroartemisinic acid to epimeric 3-hydroxyartemisinic acid by using suspension culture cell of A.annua.The results indicate that cells of A.annua have the ability to hydroxylate dihydroartemisinic acid region selectively.

Biotransformation of furannoligularenone by cell suspension cultures of Nicotiana tabacum / 中草药

Objective To investigate the biotransformation of furannoligularenone by cell suspension cultures of Nicotiana tabacum. Methods Furannoligularenone was added to the medium of the suspension cells of N. tabacum after precultured for 10 d, then they were co-cultured for another 4 d. The biotransformed products were detected with HPLC and isolated by various chromatographic methods. The chemical structures of biotransformed products were elucidated on the basis of their physicochemical pro-perties and spectroscopic data. Otherwise, the influence of co-cultured time on conversion ratio was investigated either. Results The substrate, furannoligularenone (Ⅰ), was successfully biotransformed by N. tabacum cultured cells. Two products, 3-oxo-eremophila-1, 7(11)-dien-12, 8-olide (Ⅱ) and 3-oxo-8-hydroxyeremophila-1, 7(11)-dien-12, 8-olide (Ⅲ) were obtained. After co-cultured for 5 d, the mole conversion ratio of the substrate reached the highest (53.1%). Conclusion It's the first report on biotransformation of furannoligularenone by N. tabacum cultured cells.

Isolation and purification of polysaccharides from transgenic crown gall cultures of Panax quinquefolium / 中草药

Objective To investigate the polysaccharides constituents from the transgenic crown gall cultures of Panax quinquefolium. Methods The crude polysaccharides PPQ50 and PPQ70 were extracted by water-ethanol method from the crown gall of P. quinquefolium. Four polysaccharides, named PPQ50-1, PPQ50-2, PPQ70-1, and PQ70-2, were isolated and purified by DEAE Sepharose and Sephacry S-100 column chromatography. Results The homogeneity of four polysaccharides was examined by gel chromatography and polarimetry. Their molecular weights were determined by gel filtration chromatography. They were 4.7?104, 2.9?105, 7.5?104, and 1.3?105, respectively. The sugar composition was analyzed by HPLC. The polysaccharides are mainly composed of neutral sugar particularly of D-glucose. The structure elucidation of PPQ50-2 was also supported by 1H-NMR and 13C-NMR spectroscopy. Conclusion All of the four polysaccharides are first isolated and purified from the transgenic crown gall cultures of P. quinquefolium.The result shows the PPQ50-2 and PPQ70-2 are both glucans.

Primary study on HPLC fingerprint of cultured Cordyceps militaris / 中草药

Objective To establish the HPLC fingerprint of cultured Cordyceps militaris. MethodsHPLC Method was used for the determination of C. militaris on Agilent ZOBAX SB-Aq C18 column (250 mm?4.0 mm, 5 ?m), and measuring with methanol 5%—60% in 30 min as elution detective wavelength at 260 nm, injection sample of 1.0 ?L and flow rate of 1.0 mL/min. The systematic optimized design of the chromatographic conditions, such as supersonic extraction, mobile phase, and detective wavelength as well was carried out. Results Fingerprint consisted of 11 common peaks and the result of methodology determination fitted to the related standards. The evaluation of cultured C. militaris by systematically comparing chromatograms with a professional analytical software recommended by National Institute for the Control of Pharmaceutical and Biological Products has been established. Conclusion The method is accurate, simple, and useful for the quality control of cultured C. militaris.

Biosynthesis of arbutin from crown gall of transgenic Panax quinquefolium / 中草药

Objective To investigate the biosynthesis of arbutin by suspension culture of crown gall of transgenic Panax quinquefolium. Methods Hydroquinone in methanol(60 mg/mL) was added to the medium of the crown gall of P.quinquefolium after precultured for 20 d,then they were co-cultured for another 60 h.The product was isolated and purified by column chromatography and its structure was identified by physicochemical properties and spectroscopic data.The dynamic curve of biotransformation was investigated by quantative analysis of arbutin with HPLC.Results The product could be obtained from both of the culture and medium,which was isolated and elucidated as 4-hydroxyphenyl-?-D-glucopyranoside(arbutin).After co-cultured for 60 h,the mole conversion ratio of hydroquinone is 88.4%,the product contents in culture and medium are 63.69 mg/flask and 1.86 mg/flask,respectively,and the excretion ratio of arbutin reaches the highest(2.92%).Conclusion It's the first time around the world that the crown gall of transgenic P.quinquefolium is used as a biotransformation system and arbutin which shows varied pharmacological activites have been got successfully.

Somatic embryogenesis and plantlet regeneration of Aconitum coreanum / 中草药

Object To develop new resources of medicinal plan t of Aconitum coreanum (L?vl.) Rapaics by the method of plant cel l biotechnology. Methods The segments of stems, leaves, embr yonal axises, radicles, and cotyledon of A. coreanum were used a s the explants. The MS medium supplied with different auxin and cytokinin was e xamined. Results The calli were induced from explants on the MS medium supplied with 2, 4-D 1 mg/L and 6-BA 0.5 mg/L in the period of 30 d. And the calli were differentiated into green conus or globe embryo after se veral subcultures period on medium MS supplied with 2, 4-D 0.2 mg/L and 6-BA 0.5 mg/L. The adventitious buds differentiated could be developed from the so matic embryo callus into roots on medium MS supplied with IBA 0.5 mg/L, and the n the intact plantlets were obtained. Conclusion The plantle ts of A. coreanum can be obtained by the techniques of somatic embryogenesis and it is indicated that this may be a new way in developing new r esource and protecting of A. coreanum.

Cell immobilization culture of Ginkgo biloba and determination of their affecting factors / 中草药

Object To investigate the application of cell immobilization culture to producing ginkgolides in Ginkgo biloba L. Methods To immobilize the cells by polyaminoresin foam as support materials, and determine the effects of different factors. Results The results showed that 71% of immobilization ratio and 22 mg/cm 3 of cell density were obtained using P29 of high density, small bore diameter with pieces of 0.5 cm ? 0.5 cm ? 0.5 cm, 0.72 g of support matrices per bottle in 65 mL of MS medium solution supplemented with 2,4-D 8.0 mg/L+KT 0.04 mg/L+NAA 0.4 mg/L and the incoulum quantity was 200 g/L. Conclusion It shows that present method has some advantages, such as simpleness, low cost, high immobilization ratio.