[Limonoids from seeds of Azadirachta indica and their cytotoxic activity].

Eight limonoids were isolated from 95% ethanol extracts of neem(Azadirachta indica) seeds by various chromatographic methods. By comparison of their spectroscopic data with those reported in the literatures, these limonoids were determined as salannin(1), 1-detigloyl-1-isobutylsalannin(2), salannol-3-acetate(3), salannol(4), spirosendan(5), 1-detigloyloxy-3-deacetylsalannin-1-en-3-one(6), nimbin(7) and 6-deacetylnimbin(8). Compounds 2 and 5 were firstly isolated from this genus and 5 represented the only example of its type. And 6 is a new natural product. 6 showed inhibitory activity against HeLa and HL-60 cells, with IC50 of(21.61±4.37) and(27.33±5.74) µmol·L⁻¹, respectively. Both 7 and 8 mildly inhibited the growth of HeLa cells, with IC50 of (33.15±5.24) and (38.56±6.41) µmol·L⁻¹, respectively.

Hydroxyl octadecenoic acids biosynthesized by crown galls of Panax quinquefolium induced by artermisinic acid.

OBJECTIVE: To investigate the effect of artermisinic acid on the secondary metabolites production of Panax quinquefolium crown galls. METHODS: Artemisinic acid was added into the suspended cells of Panax quinquefolium crown galls and co-culture for two days. Products were isolated with chromatographic method. RESULTS: Three hydroxyl octadecenoic acids [9,12,13-trihydroxy-10-octadecenoic acid (1), 11,12,13-trihydroxy-9-octadecenoic acid (2) and 11-hydroxy-12,13-epoxy-9-octadecenoic acid (3)] were isolated from crown galls of Panax quinquefolium. CONCLUSION: Artermisinic acid as one of the new type of phytohormones that might induce the production of 13-lipoxygenases in crown galls of Panax quinquefolium.

Structure characterization and antioxidant activity of a novel polysaccharide isolated from pulp tissues of Litchi chinensis.

A novel polysaccharide (LCP50S-2) with antioxidant activity was isolated from Litchi chinensis Sonn. The structure of LCP50S-2 was elucidated on the basis of physicochemical and instrumental analyses, and its average molecular weight was determined by gel permeation chromatography to be 2.19 × 10(2) kDa. The backbone of LCP50S-2 was composed of (1→3)-linked ß-L-rhamnopyranosyl residues, (1→4)-linked α-D-xylopyranosyl residues, (1→4)-linked ß-D-glucopyranosyl residues, and (1→4)-linked α-D-glucopyranosyl residues which branched at O-6. The two branches consisted of α-L-arabinopyranosyl residues and (1→6)-linked ß-D-galactopyranosyl residues terminated with α-L-arabinopyranosyl residues, respectively. In the in vitro antioxidant assay, LCP50S-2 was found to possess DPPH radical-scavenging activity and hydroxyl radical-scavenging activity with IC(50) values of 220 and 266 µg/mL, respectively.

Two new compounds from transgenic Panax quinquefolium.

A new ceramide and a new poly-hydroxyl octadecenoic acid were isolated from transgenic crown galls of Panax quinquefolium. Their structures were elucidated as (2S, 3S, 4R, 20E)-2-[(2'R)-2'-hydroxyl-palmitoyl-amino]-20-hexacosene-1, 3, 4-triol (1) and 12, 13, 15-trihydroxy-9-octadecenoic acid (2) respectively on the basis of spectroscopic and chemical methods.

[Optimization of culture conditions for crown gall of Panax quinquefolium and the content determination of their polysaccharides].

OBJECTIVE: To optimize the culture conditions and determine the content of polysaccharides in crown gall of Panax quinquefolium. METHODS: The orthogonal design was used for the optimization of culture condition (such as media, inoculum amount, pH and the content of inorganic elements in media) and their effects on the polysaccharides content. RESULTS: (1) The crown gall tissue could grow and produce polysaccharides in MS (Murashige & Skoog) solid medium without any hormones; (2) The time-dependent regularity of the growth amount and polysaccharides content of the crown gall tissue in MS solid medium was approached: polysaccharides content was the highest in 21 d and biomass achieved to the greatest in 24 d; (3) The polysaccharides content and growth amount were the highest when pH 5.6 of media; (4) The inoculation of 4 - 7 g (FW/flask) was comparatively advantage to the growth of the crown gall, but no effect on polysaccharides content. CONCLUSION: This method can be used to accumulate the high contents of polysaccharides in crown gall cultures of P. quinquefolium.

[Comparative studies on the amount of ginsenosides between the crown galls and Chinese medicinal materials of Panax quinquefolium].

OBJECTIVE: To provide theoretical basis for the production of ginsenosides by using modern biotechnologies, the comparative studies of the amount of ginsenosides between the crown galls and Chinese medicinal materials of Panax quinquefolium were carried out. METHODS: Total ginsenosides were determined by spectrophotometry, and the contents of ginsenoside Rb1, Rb3, Rc, Re were determined by HPLC. RESULTS: (1) The total ginsenosides in the crown galls of Panax quinquefolium (aftert 27 days' cultivation) was almost as much as that of Chinese medicinal material of Panax quinquefolium. (2) The contents of ginsenoside Rb1, Re in the crown galls were about half of that in Chinese medicinal materials of Panax quinquefolium, and ginsenoside Rc was about 80%. But the amount of ginsenoside Rb3 in the crown galls of Panax quinquefolium was much more than that in Chinese medicinal materials of Panax quinquefolium as almost 15 times higher. CONCLUSION: The crown galls of Panax quinquefolium may be a new potential resource for large-scale production of ginsenosides.

Biotransformation of thymol by hairy roots of transgenic Polygonum multiflorum.

OBJECTIVE: The exogenous substrate, thymol, was firstly biotransformed by using suspension hairy roots of transgenic Polygonum multflorum, and its biotransformed situation was also investigated. METHODS: After five days co-cultivated period, the transformed product was isolated by Thin Layer Chromatograph and Column Chromatograph, with the structure elucidated by physic-chemical methods and spectra data. Meanwhile, the time course of biotransformation (T-C) for thymol was also measured by HPLC to illuminate its bio-transformed situation. RESULTS: The glycosylated product, namely DMP, was isolated and purified, which structure was determined as 5-methyl-2-(1-methylethyl) phenyl- beta-D-glucopyranoside. And the distribution of DMP in the medium or culture was varied in different co-cultivated periods, and for five days co-cultivated period, it mainly existed in the medium. CONCLUSION: The hairy roots of Polygonum multiflorum were able to convert the aromatic exogenous substrate, thymol, into its glycoside. Furthermore, the time course indicated the relationship between DMP and co-cultivated period.

[Biotransformation of furannoligularenone by hairy root cultures of Polygonum multiflorum].

OBJECTIVE: To investigate the biotransformation of furannoligularenone by hairy roots of Polygonum multiflorum. METHODS: Furannoligularenone was added to the medium of hairy roots of P. multiflorum after precultured for 9 days, then they were co-cultured. The products were isolated and identified on the basis of their physical chemical properties and spectroscopic data. The T-C curve of biotransformation was investigated with HPLC. RESULTS: The hairy roots of P. multiflorum transformed the substrate to two products, 3-oxo-eremophila-1,7(11)-dien-12,8-olide(II) and 3-oxo-8-hydroxy-eremophila-1,7(11)-dien-12,8-olide(III). After co-cultured for 3 days, the mole conversion ratio of the substrate reached the highest (27.2%). CONCLUSION: It's possible to biotransform furannoligularenone by hairy roots of P. multiflorum.

[Study on HPEC fingerprint of cultured Cordyceps militaris].

OBJECTIVE: To investigate the High Performance Capillary Electrophoresis (HPCE) fingerprint of cultured Cordyceps militaris (L.) Link. METHODS: Separation was performed on a 50 cm x 75 microm uncoated capillary with 0.5 mmol/L borate solution (pH 9. 18) as HPEC buffer. The run voltage was 20 Kv, temperature 25 degrees C and the DAD detection was set at 254nm. RESULTS: Fingerprint consisted of 11 common peaks. The validation of methods was satisfied with the requirements for SFDA's technical regulations. CONCLUSION: The method was accurate and simple and suitable to the quality control of cultured Cordyceps militaris (L.) Link.

[Studies on biotransformation of arbutin by 4-hydroxy phenol in hairy root of Polygonum multiflorum].

OBJECTIVE: To study the biotransformation of arbutin by 4-hydroxy phenol in hairy root of Polygonum multiflorum. METHOD: 4-hydroxy phenol was used as substrate, the standard curve was made by HPLC, and the influences of the co-culture time, the concentration of substrate added and the volume of culture flasks on biotransformation of arbutin were measured by the index of the production yield and transform rate of arbutin. RESULT: Arbutin could be detected from both of the cultures and medium. The correlation curve of arbutin: Y = 440740X - 1.473 (r = 0.9997). The production yield (2.22 g x L(-1)) and conversion ratio (81.45%) of arbutin reached the maximum amount as co-culture time at 72 h, substrate added in medium for 1100 mg x L(-1). Furthermore a large-scale culture of 3 L was also successful in our experiment. CONCLUSION: It was firstly to biosynthesis arbutin in hairy root of P. multiflorum. The production yield and trasfer rate of arbutin were increased largely. And large-scale production (3 L culture flask) of arbutin was achieved in the experiment and it would be valuable for the industrial production of arbutin by biotechnological method in the future.

[Study on supension culture and quantitative determination of total ginsenosides from the crown gall of Panax quinquefolium].

The influences of different cultural node on the crown gall culture of Panax quinquefolium and its ginsenosides contents were studied. The faster growth rate was obtained in suspension culture. And the dry weight (DW) of crown gall and the accumulation of total ginsenoside content reached 0. 776 g DW/flask and 19.4 mg/flask, which were 111% and 64% higher than solid culture, respectively. The results also indicated that the utilization (by percentage) of sucrose was 91.8%. The investigation provided useful information for following researches on fermentation culture and large-scale production of ginsenosides.

[Research progress in Laggera medicinal plants].

This paper reviewed the worldwide research progresses of the genus Laggera both on phytochemical and pharmacological work in the past few decades. The main secondary metabolites of this genus are proved to be sesquitepenoids, flavonoids and phenolic acids. Phamacological investigations revealed that the certain extracts of some Laggera species possess significant bioactivities on anti-inflammation, anti-tumor and anti-viral infection. This review afforded the comprehensive description of the active components as to provide useful references to elucidate their historical clinical application on upper respiratory infection, influenza, parotitis, and recurrent herpes viral infection.

The growth characteristics and ginsenosides isolation of suspension-cultured crown gall of Panax quinquefolium.

The growth characteristics and ginsenosides isolation of the suspension-cultured crown gall of Panax quinquefolium were studied. The result showed that the maximum biomass of cultures was 18.6 g/L (dry weight) and the content of ginsenosides reached its maximum level of 620.4 mg/L on the 27th day. The utilization rates of sugar, phosphorus, nitrogen in NH4+ and nitrogen in NO3- were 91.8%, 100%, 81% and 97%, respectively. Four compounds were isolated from the suspension-cultured crown gall and their structures were elucidated as pseudoginsenoside F11 (I), ginsenoside Rd (II), ginsenoside Rb1 (llI) and ginsenoside Rb3 (IV).

[Study on the culture of crown gall from Panax quinquefolium and the prouction of its secondary metabolites--Ginsenosides Re and Rg1].

It was clearly demonstrated that T-DNA of Agrobacterium tumefeciens Ti plasmid was integrated into the cells of crown gall in our experiment. This paper reported the influences of some kinds of physical-chemistry factors on the growth of crown gall of Panax quinquefolium and the production of its main active compounds--ginsenoside Re and ginsensoside Rg1. The results showed that White medium was the best one for ginsensoside Rg1 accumulation (0.095%) among the six media, but ginsensoside Re accumulation (0.194%) was the highest on the MS medium; The highest contents of ginsensoside Re (0.147%) and ginsensoside Rg1 (0.061%) were on the culture 36d and 32d after innoculum respectively; The optimum pH was 5.6 for ginsensoside Rg1 synthesis(0.054%), and 5.8 for ginsensoside Re synthesis(0. 184% ); The contents of ginsensoside Re and ginsensoside Rg1 was the highest in the inoculum of 4 g and 2 g/flask (FW) respectively. The result also indicated that the concentration of inositol in 0.05 g/L could obviously promote ginsensoside Re synthesis (0.182%), and in 0.30 g/L for ginsensoside Rg1 (0.055%).