[Chemical constituents from fruiting bodies of Tremella sanguinea].

The chemical constituents from the fruiting bodies of Tremella sanguinea were separated and purified by column chromatography on silica gel,ODS,Sephadex LH-20,and RP-HPLC. The structures of the isolated compounds were identified on the basis of physicochemical properties and spectroscopic data analysis,as well as comparisons with the data reported in the literature. Sixteen compounds were isolated from the 90% ethanol extract of the fruiting bodies of T. sanguinea,which were identified as( 22 E)-5α,8α-epidioxy-24-methyl-cholesta-6,9( 11),22-trien-3ß-ol( 1),( 22 E)-5α,8α-epidioxyergosta-6,22-dien-3ß-ol( 2),cerevisterol( 3),ergosta-7-ene-3ß,5α,6ß-triol( 4),( 22 E)-6ß-methoxyergosta-7,22-diene-3ß,5α-diol( 5),ergosta-7-en-3ß-ol( 6),4-hydroxy-methylincisterol( 7),2-pyrrolidone( 8),nicotinamide( 9),1-( 3-indolyl)-3-dihydroxypropan-1-one( 10),yangambin( 11),linoleic acid( 12),( 9 Z,12 Z,15 Z)-2,3-dihydroxypropyl octadeca-trienoate( 13),( 9 Z,12 Z)-2,3-dihydroxypropyl-octadeca-dienoate( 14),crypticin B( 15)and 3-phenyllactic acid( 16). All compounds were isolated from T. sanguinea for the first time. Except for compounds 6,9 and 12,the remained compounds were isolated from the genus Tremella for the first time.

[Identification of four Armillaria strains and their effects on quality and yield of Gastrodia elata f. glauca].

In order to improve the quality and yield of Gastrodia elata f. glauca,determine the suitable Armillaria strains for the accompanying experiment in Xiaocaoba,Yiliang,four Armillaria strains were selected. They were used for G. elata cultivation,and the gene sequence,r DNA-ITS,ß-tubulin and EF1-α of four Armillaria strains,were compared and analyzed. The yield was mesured in November which was based on previous laboratory research. The tubers were washed and steamed,then dried and powdered. The content of gastrodin and p-hydroxybenzyl alcohol was determined by UPLC,the polysaccharide was determined by phenol-concentrated sulfuric acid method. The results showed that the strains M1,M2,M3 and M4 were Armillaria gallica group but there were differences in the yield and active ingredient content when they were cultivated with the same G. elata. The yield of G. elata( Jian Ma) was the lowest when cultivated with Armillaria strain M3,but it was not the same when used M1,0. 981 kg·m-2,the highest yield in the four stains.The content of gastrodin was 0. 581%,the total content of gastrodin and p-hydroxybenzyl alcohol was 0. 595%,when accompanied with M1 strains. It was higher than other strains. The content of G. elata polysaccharide was 2. 132%,which was similar to the content of M3 strain,higher than that of M2 and M4 strain. Selecting phylogenesis of Armillaria strians,the content of active ingredient,and the yield as indicators,it was concluded concerned that the M1 strain was the best of four strains. The results will provide a theoretical basis and guidance for higher yield and quality in cultivation of G. elata in Yiliang.

[Antitumor activity and structure-activity relationship of seven lanostane-type triterpenes from Fomitopsis pinicola and F. officinalis].

Seven lanostane-type triterpenes including fomitopsin C(1),3-keto-dehydrosulfurenic acid(2),dehydroeburiconic acid(3),3-acetyloxylanosta-8, 24-dien-21-oic acid(4),pinicolic acid A(5),trametenolic acid B(6),and eburicoic acid(7),were isolated from the fruitbodies of Fomitopsis pinicola and F. officinalis. In vitro assay, all compounds were evaluated against MCF-7, HeLa, HepG2 and A549 cells lines using the MTT assay and the structure-activity relationship of antitumor activity was discussed. The results showed that the seven compounds were more sensitive to MCF-7 cells.The IC50 value for MCF-7 was 2<5<4<1<3<6<7. H22 tumor mouse model was used to assay compounds 2, 3, 4 and 5 in vivo. Compounds 2 and 4 had obvious effect and the necrosis area and measurement were positively correlated. The results showed that compounds 2, 4 and 5 had significant antitumor activities at a dose of 20 mg•L⁻¹ with 65.31%, 56.71%, 58.72% suppression, respectively, approaching to CTX group with 69.19% suppression in subcutaneous H22-implanted mice.The results showed that these compounds had significant against the expression of VEGF, cytokines IL-4 and IFN-γ tumor, additionally, the structure-activity relationship of lanostane-type triterpenes indicated that the acetoxyl or carbonyl at C-3 and hydroxy at C-15 can enhance the antitumor activity.

[Antitumor Effect of Solid State Fermentation Powder of Inonotus hispidus on H22 Bearing Mice].

Objective: To study the inhibitory effect and the mechanism of solid fermentation powder（ SFP） of Inonotus hispidus on H22 bearing mice. Methods: The H22 transplanted models of mice were established by H22 ascites tumor. 90 mice were divided into nine groups as control,model,cytoxan（ CTX,20 mg / kg） positive control,high-,mid-,low-dose（ 1 000,750,and 500 mg / kg） of corn SFP of Inonotus hispidus,and high-,mid-,low-dose（ 1 000,750,and 500 mg / kg） of sorghum SFP of Inonotus hispidus. On the day after being inoculated with H22 tumor cells, mice were given SFP of Inonotus hispidus once daily for consecutive 11 d. And then the increaseing of the tumor inhibitory rate,body weight,thymus,spleen,and liver indexes were calculated; and the histopathologic changes of tumor of H22 bearing mice were observed. The activities of MDA,SOD and CAT, the contents of GSH-Px in liver homogenates and blood serum,and the number of IL-2,VEGF,and IFN-γ in blood serum were detected. Results: The treatment with SFP of Inonotus hispidus significantly increased the body weight,thymus index,spleen,and liver indexes. The histopathologic study showed that the necrosis in H22 cells was induced,and the hepatic SOD,CAT,GSH-Px activities were increased,the MDA in liver homogenates and blood serum were decreased, the serum levels of IL-2 and IFN-γ were elevated, and the VEGF level was reduced. Conclusion: SFP of Inonotus hispidus have an antitumor effect on H22 bearing mice,and the possible mechanism may be due to its antioxidant activity,improving immunofunction,and antiangiogenetic action.

[Advance in studies on chemical constituents and pharmacological activity of lichens in Usnea genus].

OBJECTIVE: To summarize the studies on chemical constituents and pharmacological activities of lichens of Usnea genus. METHOD: A systematic literature survey was conducted to classifiy and summarize chemical constituents of lichens of Usnea genus, and sum up current studies on main pharmacological activities of lichens of the genus. RESULT: Lichens of Usnea genus contained multiple chemical constituents, primarily including mono-substituted phenyl rings, depsides, anthraquinones, dibenzofurans, steroids, terpenes, fatty acids and polysaccharides, with such biological activities as antitumor, antibacterial, anti-inflammation, anti-oxidation and antithrombosis. CONCLUSION: This essay provides reference for further studies and development of lichens of Usnea genus.

[Chemical constituents from Usnea longgisima, a traditional mongolian medicine].

OBJECTIVE: To study the chemical constituents of the whole lichen of Usnea longissima. METHOD: The compounds were separated by silica gel, Sephadex LH-20 chromatography and high performance liquid chromatography (HPLC). The structures of the compounds isolated were identified by physico-chemical properties and spectral analysis. RESULT: Ten compounds were isolated and their structures were identified as (4aR,9bS)-2,6-diactyl-3,4a,7,9-tetrahydroxy-8,9b-dimethyl-1-oxo-1,4,4a, 9b-tetrahydrodibenzo [b,d]furan (1), (+)-usnic acid (2), orcinol (3), 18R-hydroxydihydroalloprotolichensterinic acid (4), 5, 8-epidioxy-5alpha, 8alpha-ergosta-6, 22E-dien-3beta-ol (5), ethyl everninate (6), arabitol(7), apigenin 7-O-beta-D-glucuronide (8), 3-hydroxy-5-methoxy-2-methylbenzoic acid(9), friedelin(10). CONCLUSION: Compound 1 was a new compound. Compound 8 was isolated from genu Usnea for the first time and compounds 3, 4 and 7 were isolated from U. longissima for the first time.

[Chemical constituents from Lyophyllum decastes].

The chemical constituents from the fruiting bodies of Lyophyllum decastes (Fr.) Singer were studied in this paper. Thirteen compounds were isolated and purified by column chromatographies on silica gel and Sephadex LH-20. Their structures were identified by MS and NMR data analysis as adenosine (1), 2R, 3S, 4S, 8E)-2-[(2'R)-2-hydroxyheneicosanoylamino]-8-octadecene-1, 3, 4-triol (2), (2R, 3S, 4S, 8E)-2-[(2'R)-2-hydroxypentacosanoylamino]-8-octadecene-1, 3, 4-triol (3), nicotinic acid (4), (4E, 8E) -2-N-2-hydroxytetracosanoyl-1-O-beta-D-glycopyranosyl-9-methyl-4, 8-sphingadienine (5), D-mannitol (6), ergosteryl-3-O-beta-D-glucopyranoside (7), tuberoside (8), (2R, 3S, 4S, 8E)-2-[(2'R)-2-hydroxybehenoylamino]-8-octadecene-1, 3, 4-triol (9),(2R, 3S, 4S, 8E)-2-[(2'R) -2-hydroxytricosanoylamino] -8-octadecene-1, 3, 4-triol (10), (22E, 24R)-ergosta-7, 22-dien-3beta, 5alpha, 6beta-triol (11), (22E, 24R)-ergosta-5, 7, 22-trien-3beta-ol (12), and 5alpha, 8alpha-epidiory-(22E, 24R)-ergosta-6, 22-dien-3beta-ol (13), respectively. All the above compounds are first obtained from the mushroom and compounds 2-10 are reported to be obtained from the Lyophyllum for the first time.

Antioxidant hispidin derivatives from medicinal mushroom Inonotus hispidus.

Two natural antioxidants, named inonotusin A (1) and B (2), were isolated from the methanolic extract of the fruit bodies of Inonotus hispidus, together with (E)-4-(3,4-dihydroxyphenyl)but-3-en-2-one (3), hispidin (4) and 3,4-dihydroxybenzaldehyde (5). Their structures were identified by means of extensive NMR and MS data analysis. Compounds 1, 2 and 4 exhibited significant scavenging activity against the 2,20-azinobis(3-ethylbenzhiazoline-6-sulfonate) (ABTS) radical cation. Compound 1 also showed moderate cytotoxicity against a human breast carcinoma cell line (MCF-7) with IC(50) values of 19.6 µM.

Antitumor components from Naematoloma fasciculare.

The bioassay-guided fractionation of MeOH extract from Naematoloma fasciculare afforded a petroleum ether fraction (NFPF) and four known compounds, which showed good antitumor activities to inhibit MCF-7 cell line proliferation in vitro and tumor growth in H(22) implanted mice in vivo. In addition, a number of unsaturated aliphatic acids were identified in NFPF by GC analysis. These results showed that NFPF inhibits tumor growth through the activity of unsaturated aliphatic acids together with two active compounds, ergosterol peroxide (1: 62.17 mg/g in NFPF) and ergosterol (2: 3.13 mg/g in NFPF), and indicate the potential utility of NFPF as an antitumor drug.

Structural elucidation of a polysaccharide from Flammulina velutipes and its immunomodulation activities on mouse B lymphocytes.

A novel polysaccharide FVPB2 was purified from fruiting bodies of Flammulina velutipes. Its structure was elucidated by monosaccharide composition and methylation analyses, UV-Visible and FTIR spectroscopy as well as NMR. FVPB2 was a homogeneous heteropolysaccharide (molecular weight ~ 1.50 × 104 Da) containing D-galactose, D-mannose, L-fucose, and D-glucose at molar ratio of 1.9:1.2:1:2.5. In vitro immunomodulatory studies showed FVPB2 induced proliferation of mouse spleen lymphocytes in a dose-dependent manner. The levels of IgM and IgG, secreted by B cells, increased after FVPB2 treatment. So FVPB2 has potential to be a new important immunomodulatory nutraceutical.

Structural Characterization and Immunological Activities of a Novel Water-Soluble Polysaccharide from the Fruiting Bodies of Culinary-Medicinal Winter Mushroom, Flammulina velutipes (Agaricomycetes).

A water-soluble polysaccharide, designated FVPA2, was isolated from the fruiting bodies of Flammulina velutipes using DEAE Sepharose Fast Flow and gel-permeation chromatography. Its structure was elucidated by monosaccharide composition and methylation analysis, ultraviolet, Fourier transform infrared spectrometry, and nuclear magnetic resonance spectroscopy. Results showed that FVPA2 was a homogeneous heteropolysaccharide containing galactose, fucose, and mannose in a molar ratio of 5:1:1. High-performance liquid chromatography indicated its molecular weight as 3.4 × 104 Da. FVPA2 also has a repeating unit. In vitro immunomodulatory studies showed that Raw264.7 cells were stimulated to secret nitric oxide upon administration of 200-500 µg/mL FVPA2. FVPA2 also stimulated the proliferation of mouse spleen lymphocytes and B lymphocytes.

Memory enhancing and neuroprotective effects of selected ginsenosides.

The effects of ginsenosides Rg3(R), Rg3(S) and Rg5/Rk1 (a mixture of Rg5 and Rk1, 1:1, w/w), which are components isolated from processed Panax ginseng C.A. Meyer (Araliaceae), on memory dysfunction were examined in mice using a passive avoidance test. The ginsenosides Rg3(R), Rg3(S) or Rg5/Rk1, when orally administered for 4 days, significantly ameliorated the memory impairment induced by the single oral administration of ethanol. The memory impairment induced by the intraperitoneal injection of scopolamine was also significantly recovered by ginsenosides Rg3(S) and Rg5/Rk1. Among the three ginsenosides tested in this study, Rg5/Rk1 enhanced the memory function of mice most effectively in both the ethanoland scopolamine-induced amnesia models. Moreover, the latency period of the Rg5/Rk1-treated mice was 1.2 times longer than that of the control (no amnesia) group in both models, implying that Rg5/Rk1 may also exert beneficial effects in the normal brain. We also evaluated the effects of these ginsenosides on the excitotoxic and oxidative stress-induced neuronal cell damage in primary cultured rat cortical cells. The excitotoxicity induced by glutamate or N-methyl-D-aspartate (NMDA) was dramatically inhibited by the three ginsenosides. Rg3(S) and Rg5/Rk1 exhibited a more potent inhibition of excitotoxicity than did Rg3(R). In contrast, these ginsenosides were all ineffective against the H2O2- or xanthine/xanthine oxidase-induced oxidative neuronal damage. Taken together, these results indicate that ginsenosides Rg3(S) and Rg5/Rk1 significantly reversed the memory dysfunction induced by ethanol or scopolamine, and their neuroprotective actions against excitotoxicity may be attributed to their memory enhancing effects.

Improvement of memory by dieckol and phlorofucofuroeckol in ethanol-treated mice: possible involvement of the inhibition of acetylcholinesterase.

Phlorotannins, the polyphonic compounds found in brown Eisenia and Ecklonia algae, have several pharmacologically beneficial effects such as anti-inflammation. In addition, our recent data show that these compounds may improve the cognitive functions of aged humans suggesting the potential ability to enhance memory in several neurodegenerative disorders. To examine the experimental hypothesis that two effective components of Ecklonia cava, dieckol and phlorofucofuroeckol (PFF), have memory-enhancing abilities, both were administered orally to mice before a passive avoidance test. The repeated administration of either dieckol or PFF dose-dependently reduced the inhibition of latency by the administration of ethanol. To investigate the mode of memory-enhancing actions, the levels of major central neurotransmitters in three different regions (striatum, hippocampus, and frontal cortex) of the mouse brain were measured. The levels of some of the neurotransmitters were significantly changed by ethanol. Both dieckol and PFF altered the levels of some neurotransmitters modified by the ethanol treatment. It is noteworthy that both dieckol and PFF increased the level of acetylcholine, and they exerted anticholinesterase activities. Overall, the memory-enhancing abilities of dieckol and PFF may result from, at least in part, the increment of the brain level of acetylcholine by inhibiting acetylcholinesterase.

A new antioxidant pyrano[4,3-c][2]benzopyran-1,6-dione derivative from the medicinal mushroom Fomitiporia ellipsoidea.

A new pyrano[4,3-c][2]benzopyran-1,6-dione derivative antioxidant, named phelligridin K (1), was isolated from the methanol extract of the fruiting bodies of Fomitiporia ellipsoidea, together with the known compounds, (E)-4-(3,4-dihydroxyphenyl)but-3-en-2-one (2), hispidin (3), inoscavin C (4), and inonoblin B (5). Their structures were elucidated by means of extensive NMR and MS data analysis. Compounds 1, 4 and 5 exhibited significant scavenging activity against the ABTS radical cation.

Isolation and identification of a new benzofuranone derivative from Usnea longissima.

A new benzofuranone derivative, ethyl 2-(3,3-bis(7-acetyl-4,6-dihydroxy-3,5-dimethylbenzofuran-2-yl)acryloyl) (1), together with seven known compounds (2-8), was isolated from Usnea longissima. Their structures were elucidated by using spectroscopic methods, including 1D and 2D NMR experiments, UV, IR, EI-MS and HR-ESI-MS analysis.