Further studies on the hepatoprotective effect of Antrodia camphorata in submerged culture on ethanol-induced acute liver injury in rats.

To further understand the hepatoprotective activity of Antrodia camphorata in living systems and the possible mechanisms of this protection, the effects of fractions from A. camphorata in submerged culture on the liver and its antioxidative system in acute ethanol intoxicated rats were investigated. The results showed that the ethanolic extract (Fr-I) of A. camphorata was the most effective in the prevention of ethanol-induced acute liver injury and free radical generation in rats. The ethanolic extract administrated prior to ethanol significantly prevented the increase in serum levels of hepatic enzyme markers such as aspartate aminotransferase and alanine aminotransferase. It also normalised the increase of hepatic malondialdehyde concentration and the decrease of glutathione levels in the liver. Moreover, Fr-I improved the ethanol-induced decrease of hepatic glutathione peroxidase and reductase activities. On the basis of these results, the ethanolic extract of A. camphorata may exert its hepatoprotective activity by up-regulating GSH-dependent enzymes and inhibiting free radical formation in the liver.

Paclitaxel production using co-culture of Taxus suspension cells and paclitaxel-producing endophytic fungi in a co-bioreactor.

The co-culture of the suspension cells of Taxus chinensis var. mairei and its endophytic fungi, Fusarium mairei, in a 20-L co-bioreactor was successfully established for paclitaxel production. The co-bioreactor consists of two-unit tanks (10 L each) with a repairable separate membrane in the center, culturing Taxus suspension cells in one tank and growing fungi in another. By optimizing the co-culture conditions, there was a desirable yield of paclitaxel in Taxus cell cultures. The Taxus cell cultures by co-culture produced 25.63 mg/L of paclitaxel within 15 days; it was equivalent to a productivity of 1.71 mg/L per day and 38-fold higher than that by uncoupled culture (0.68 mg/L within 15 days). The optimum conditions for co-culture in the co-bioreactor were: B5 medium, inoculating fungi when Taxus cells had grown for 5 days in the co-bioreactor, hydrophilic separate membrane in the center of the co-bioreactor, and air flow rate of 1:0.85 v/v/m in fungus cultures.

Phoxalone, a novel macrolide from Sorangium cellulosum: structure identification and its anti-tumor bioactivity in vitro.

A novel macrolide, isolated from the myxobacteria Sorangium cellulosum WXNXJ-C, was identified as 1,7,12,13-tetrahydroxy-14-methoxy-8,10-dimethyl-6-phenyl-5,15-dioxa-bicyclo[9.3.1]pentadecan-4-one, "Phoxalone". It had minimum IC(50) values of 0.24, 6.9, 10.3, 0.98 and 4 microg/ml, respectively, against tumour cell lines: B16, Bel7402, H446, MCF-7, and SGC7901. In addition, it had less cytotoxicity to normal human liver L02 cell lines (286 microg/ml, 24 h). A cytotoxic bioactivity study on H446 cell line in vitro suggested that Phoxalone arrested the mitosis in the G2/M phase.

A myxobacterium strain Sorangium cellulosum AHB125 producing epothilone B and other anticancer substances.

A myxobacterium strain AHB125 belonging to genus Sorangium cellulosum was isolated from Anhui area in China and identified with morphological analysis by electron microscopy and phase contrast microscope according to Bergey's manual of systematic bacteriology (8th Ed.). Its high-antitumor bioactivity metabolites was evaluated by bioassay-directed screening technique with B16 tumor cell line etc. Research results showed that it exhibited not only strong antitumor ability bioactivities and broad-spectrum antitumor abilities to B16, Bel7402, H446, SGC7901 cell lines, but also has selectivity and pertinence to B16 and SGC7901 cell lines. The compound was confirmed as epothilone B by HPLC and LC/MS analysis, compared to the epothilone B standard sample. Bioassay indicated that there were other high-bioactive substances in the metabolites.

Study on the structure-activity relationships of phoxalone and induction of apoptosis in H446 tumor cells.

A novel macrolide with dioxa and double ring pentadecanone, named as phoxalone, isolated from the culture broth of a myxobacterium Sorangium cellulosum WXNXJ-C, was well investigated. The chemical structure of Phoxalone was elucidated by spectroscopic analysis including UV, IR, (1)H NMR, MALDI-TOF-MS, and LC/MS spectra. The results of cytotoxic bioactivities on human non-small lung cancer H446, in vitro, showed that not only did phoxalone express higher antitumor bioactivity than many antitumor drugs, but it also displayed less cytotoxicity to normal human liver L02 cell lines (IC(50), 286 microg mL(-1)). Further cytotoxic bioactivity study indicated that phoxalone could induce H446 cell line apoptosis in vitro. More investigation results of flow-cytometric analysis suggested that phoxalone arrested the mitosis of H446 cell line at G2/M phase. Hence, phoxalone and its derivatives or analogs would reveal huge research value and fascinating foreground.

Protective effects of mycelia of Antrodia camphorata and Armillariella tabescens in submerged culture against ethanol-induced hepatic toxicity in rats.

The hepatoprotective effects of the mycelia of Antrodia camphorata and Armillariella tabescens were evaluated in vivo using acute ethanol-intoxicated rats as an experimental model. Animals were orally treated with Antrodia camphorata (0.5 or 1.0 g/kg b.w.) or Armillariella tabescens (0.5 or 1.0 g/kg b.w.) for 10 days whereas controls received vehicle only. At the end of the experimental 10-day period, the animals were administered by gavage with an acute ethanol dose of 5.0 g/kg b.w. diluted in deionized water (6:4, v/v) and sacrificed at 18 h after ethanol administration. The degree of protection was measured by using biochemical parameters like serum transaminases (AST and ALT), alkaline phosphatase (ALP), bilirubin. Meanwhile, the histopathological studies were carried out to support the above parameters. Administration of Antrodia camphorata or Armillariella tabescens markedly prevented ethanol-induced elevation of levels of serum AST, ALT, ALP, and bilirubin comparable with standard drug silymarin.

Induction of apoptosis in SGC-7901 cells by polysaccharide-peptide GFPS1b from the cultured mycelia of Grifola frondosa GF9801.

The biological function of GFPPS1b, a novel polysaccharide-peptide isolated from cultured mycelia of Grifola frondosa GF9801, was well investigated. GFPS1b has anti-tumor activity and can significantly inhibit the proliferation of SGC-7901 cells, whereas slightly influences the growth of human normal liver cell line L-02. When treated with GFPS1b, SGC-7901 cells showed typical apoptotic morphological features such as the loss of villus and appearance of apoptotic bodies on the cell surface, volume reduction, and chromatin condensation, by scanning electron microscopy (SEM) and fluorescent microscopy (Hoechst 33342). The results of flow cytometry analysis and annexin V-PI assay showed that the SGC-7901 cell cycle was arrested in the G(2)/M phase, the subdiploid peak of DNA characteristic of apoptotic was also observed, and the apoptosis ratio was about 15.08%. DNA isolated from SGC-7901 cells cultured with GFPS1b showed a typical DNA 'ladders' of apoptosis in agarose gel electrophoresis. Further investigation results showed that the apoptotic machinery of SGC-7901 induced by GFPS1b was associated with drop in mitochondrial trans-membrane potential, upregulation of Bax, downregulation of Bcl-2, and activation of caspase-3. Our finding suggests that GFPS1b could suppress SGC-7901 cell growth and reduce cell survival via arresting cell cycle and inducing apoptosis of tumor cells.

[Inhibitory effect of weikangfu recipe on growth of mouse S180 tumor and its apoptotic induction].

OBJECTIVE: To study the growth inhibition of Weikangfu recipe on S180 tumor and its apoptotic induction. METHOD: S180-bearing mice were orally administrated with different dosages of Weikangfu recipe, and the growth inhibition was evaluated; apoptotic cells induced were detected by flow cytometry and DNA agarose gel electrophoresis. RESULT: Weikangfu recipe showed significant inhibition on the growth of S180 tumor in a dose-dependent manner, compared with the control group. From apoptotic analyses, Weikangfu recipe induced a dose-dependent apoptosis of S180 tumor cells and arrested the cell cycle distribution at G0-G1 phase. At the same time, the up-regulation of p53 and bax and down-regulation of bcl-2 were observed in S180 tumor cells of the treated groups. CONCLUSION: Our findings demonstrate that Weikangfu recipe can significantly inhibit the growth of S180 tumor and induce apoptosis through expression alteration of p53, bax and bcl-2.

[Studies on active substance of anticancer effect in Polygonum cuspidatum].

OBJECTIVE: To supply the scientific basis of research and development of the medicinal value of Polygonum cuspidatum. METHODS: One composition was isolated from the roots of Polygonum cuspidatum by cytotoxicity based fractionation and identified by HPLC-MS, UV scanning and IR. The inhibition and morphology of L-02, Hep G2, SHZ-888, MCF-7, MCF-7/ADM cells growth caused by this composition was determined by MTT assay and HE dyeing. RESULTS: This composition was identified as trans-and cis-resveratrol. It could specifically inhibit proliferation of many cancer cells but not human normal liver cell. We investigated the cytotoxicity of resveratrol to adriamycin-resistant MCF-7 cell in virtro. CONCLUSION: Resveratrol is a new anticancer composition which is less toxicity and higher efficiency in Polygonum cuspidatum.

Analysis of the resveratrol-binding protein using phage-displayed random peptide library.

Resveratrol, a plant polyphenol, is found in significant amounts in the skin of grapes and in some traditional herbs. It is reported to exert different biological activities, such as inhibiting lipid peroxidation, scavenging free radicals, inhibiting platelet aggregation, and anticancer activity. In order to screen the resveratrol-binding proteins, we synthesized biotinylated resveratrol, purified by liquid chromatography and immobilized it into streptavidin-coated microplate wells. 3-(4,5-Demethylthiazol-)-2,5-diphenyl tetrazolium bromide assay showed little change in the anticancer activity of biotinylated resveratrol in vitro. A random library of phage-displayed peptides was screened for binding to immobilized resveratrol to isolate resveratrol-binding proteins. Several peptides were found to bind to resveratrol specifically, which was proven by enzyme-linked immunosorbent assay. Through amino acid sequence analysis of the selected peptides and human proteins using the BLAST program, the results showed that resveratrol has an affinity for various proteins such as breast cancer-associated antigen, breast cancer resistance protein, death-associated transcription factor, and human cyclin-dependent kinase. These results demonstrate that our study provides a feasible method for the study of binding proteins of natural compounds using a phage-displayed random peptide library.

Effects of cerium on growth and physiological mechanism in plants under enhanced ultraviolet-B radiation.

Effect of cerium (Ce3+) on the growth, photosynthesis and antioxidant enzyme system in rape seedlings (Brassica juncea L.) exposed to two levels of UV-B radiation (T1: 0.15 W/m2 and T2: 0.35 W/m2) was studied by hydroponics under laboratory conditions. After 5 d of UV-B treatment, the aboveground growth indices were obviously decreased by 13.2%-44.1% (T1) and 21.4%-49.3% (T2), compared to CK, and except active absorption area of roots, the belowground indices by 14.1%-35.6% (T1) and 20.3%-42.6% (T2). For Ce+UV-B treatments, the aboveground and belowground growth indices were decreased respectively by 4.1%-23.6%, 5.2% -23.3% (Ce+T1) and 10.8%-28.4%, 7.0%-27.8% (Ce+T2), lower than those of UV-B treatments. The decrease of growth indices appeared to be the result of changes of physiological processes. Two levels of UV-B radiation induced the decrease in chlorophyll content, net photosynthesis rate, transpiration rate, stomatal conductance and water use efficiency by 11.2%-25.9% (T1) and 20.9%-56.9% (T2), whereas increase in membrane permeability and activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) by 6.9%, 22.8%, 21.5%, 9.5% (T1) and 36.6%, 122.3%, 103.5%, 208.9% (T2), respectively. The reduction of the photosynthetic parameters in Ce+UV-B treatments was lessened to 3.2%-13.8% (Ce+T1) and 4.9%-27.6% (Ce+T2), and the increase of membrane permeability and activities of antioxidant enzymes except POD in the same treatments were lessened to 2.4%, 8.4%, 6.6% (Ce+T1) and 30.1%, 116.7%, 75.4% (Ce+T2). These results indicate that the regulative effect of Ce on photosynthesis and antioxidant enzymatic function is the ecophysiological basis of alleviating the suppression of UV-B radiation on growth of seedlings. Furthermore, the protective effect of Ce on seedlings exposed to T1 level of UV-B radiation is superior to T2 level.

Responses of antioxidant enzyme and photosynthesis in rape seedling to the combined stresses of acid rain and ultraviolet-B radiation.

Effects of the simulated acid rain (AR) and ultraviolet-B (UV-B, 280-320 nm) radiation with a single or two ways simultaneously (AR + UV-B) on the antioxidant enzyme and photosynthesis of the rape seedlings were investigated by the hydroponic culture. The results of static experiment indicated that the tolerance of rape seedling to single stress (AR or UV-B) is stronger than that to dual stresses (AR + UV-B). Furthermore, the dual stresses had additive effect on catalase activity, and a synergistic effect on MDA content, net photosynthesis rate, water use efficiency as well as intercellular CO2 concentration. Meanwhile, it has an independent effect on chlorophyll content, stomatal conductance, and transpiration rate as well as membrane permeability. During 64 h restoration course, the dynamic change in the curves of physiological and biochemical indices were not identical, and none of them show a simple linear variation. According to the static and dynamic experiments, it was found that a responsive sequence of catalase activity, membrane permeability, MDA content and photosynthetic characteristics to the above-mentioned stresses was as follows: AR + UV-B > UV-B > AR.