Although the pharmacology and clinical application of water extracts of Ganoderma lucidum have been extensively documented, little is known regarding its alcohol extract. In the present study, the anti-tumor effect of an alcohol extract of Ganoderma lucidumwas investigated using MCF-7 cells. We found that the alcohol extract of Ganoderma lucidum inhibited cell proliferation in a dose-and time-dependent manner, which might be mediated through up-regulation of p21/Waf1 and down-regulation of cyclin D1. Furthermore, this compound can directly induce apoptosis in MCF-7 cells, which might be mediated through up-regulation of a pro-apoptotic Bax protein and not by the immune system. Our findings suggest that there are multiple mechanisms underlying the anti-tumor effects of Ganoderma lucidum.

Collagen extracted from pigskin was coronacharged negatively by a specifically designed device. Different charging voltages, temperatures and times were applied to prepare collagen bioelectret. The decline of the surface potential of the bioelectret under different treatment was then determined. The data showed that the surface potential was markedly varied with the charging conditions. The optimal values of three parameters for charging collagen coatings were defined as follows: voltage, 8 KV; temperature, 40◦C; time, 25 min. Treatment of the bioelectret with distillated water, or saline solution (0.9%) or culture medium induced a sharp decrease of the surface potential. In addition, we investigated the effects of the charged collagen on cell growth and intracellular calcium level of three types of cultured mammalian cell lines, including Chinese hamster ovary CHO cells, human cervix uteri tumor HeLa cells and human promyelocytic HL-60 cells. Cell growth and the intracellular calcium level were determined by MTT reagent-based assay and a fluorescent probe Fura-2, respectively. The results showed that negatively charged collagen stimulated the growth of CHO or HL-60 cell line but inhibited the growth of HeLa cell line. Furthermore, after attaching to the charged collagen, the intracellular calcium level of CHO cells increased, while that of HeLa cells decreased.

Influence of fullerene (C60) derivatives on M-MuLV reverse transcriptase activity is investigated. Two water-soluble fullerene derivatives, fullerol (C60(OH)23-24) and trimalonic acid C60 (TMA C60, C63(COOH)6) are used in the experiments and their effects on in vitro reverse transcription-polymerase chain reaction (RT-PCR) of β-actin mRNA in HeLa cells are determined. PCR products are detected by agarose gel electrophoresis. It is found that the amounts of PCR products decrease with addition of either of two fullerene derivatives to RT reaction mixture. The inhibition of fullerene derivatives is dose-dependent, and IC50 values of fullerol and TMA C60 are 0.089 and 0.039 mmol/L, respectively. The amount of PCR products obtained by direct addition of fullerol or TMA C60 to PCR are greater than those obtained by addition of the equivalent amount of fullerol or TMA C60 to RT, indicating an inhibitory effect of fullerol or TMA C60 on RT. The compensative experiment of M-MuLV reverse transcriptase shows that increasing enzyme amounts can antagonize the activity of fullerol or TMA C60. These results imply that fullerenes can inhibit M-MuLV reverse transcriptase activity, with the inhibition of TMA C60 slightly stronger than fullerol, and that their potential in treatment of diseases induced by RNA viruses such as leukemia virus needs further investigation.

Enzyme inhibition by fullerene derivatives has attracted much attention. In this communication, effects of two water-solube fullerene derivatives, fullerol and trimalonic acid C60 (TMA C60) on polymerase chain reaction (PCR) were investigated by using PCR of β-actin cDNA derived from HeLa cells as an experimental model. Both fullerol and TMA C60 were found to inhibit PCR in a dose-dependent manner. PCR was ultimately inhibited while the concentrations of each compound were not less than 0.01mM. In contrast, mannitol exerted no effects on PCR while its concentration increased up to 2mM. Compensation experiments with Thermus aquaticus (Taq) DNA polymerase revealed that both fullerol andTMAC60 inhibited the enzymatic activity of Taq DNA polymerase, and the inhibitory potency of TMA C60 was slightly greater than that of fullerol. Our data provides some novel aspects on the enzyme inhibiting activities of fullerene derivatives.