Methotrexate (MTX), a stoichiometric inhibitor of dihydrofolate reductase, is a chemotherapeutic agent for treating a variety of neoplasms. Impairment of drug import into cells and increase in drug export from cells may render cells resistant to MTX. MTX, when locally administered in a soluble form, is rapidly absorbed through capillaries into the circulatory system, which may also account for therapeutic failure in patients. To retain MTX within tumor cells for longer duration and alter its pharmacokinetic behavior, we proposed a new formulation of MTX bound to the gold nanoparticle (AuNP) that serves as drug carriers. In this study, we developed the MTX-AuNP conjugate and examined its cytotoxic effect in vitro and antitumor effect in vivo. Spectroscopic examinations revealed that MTX can be directly bound onto AuNP via the carboxyl group (-COOH) to form the MTX-AuNP complex and kinetically released from the nanoparticles. The accumulation of MTX is faster and higher in tumor cells treated with MTX-AuNP than that treated with free MTX. Notably, MTX-AuNP shows higher cytotoxic effects on several tumor cell lines compared with an equal dose of free MTX. This can be attributed to the "concentrated effect" of MTX-AuNP. Administration of MTX-AuNP suppresses tumor growth in a mouse ascites model of Lewis lung carcinoma (LL2), whereas an equal dose of free MTX had no antitumor effect. In conclusion, these results suggest that by combining nanomaterials with anticancer drugs MTX-AuNP may be more effective than free MTX for cancer treatment.

Objective. Angiogenesis plays a part in the pathogenesis of rheumatoid arthritis (RA), and nanogold inhibits the activity of an angiogenic factor, vascular endothelial growth factor (VEGF). We therefore investigated whether intraarticular delivery of nanogold ameliorates collagen-induced arthritis (CIA) in rats.Methods. Binding of 13-nm nanogold to VEGF in human RA synovial fluid (SF) and its effects on RA SF-induced endothelial cell proliferation and migration were assessed. Nanogold was administered intraarticularly to rats with CIA before the onset of arthritis. Progression of CIA was monitored by measures of clinical, radiologic, and histologic changes. In addition, the microvessel density and extent of infiltrating macrophages as well as levels of tumor necrosis factor ␣ (TNF␣) and interleukin-1␤ (IL-1␤) in the ankle joints were determined.Results. Nanogold bound to VEGF in RA SF, resulting in inhibition of RA SF-induced endothelial cell proliferation and migration. Significant reductions in ankle circumference, articular index scores, and radiographic scores were observed in the nanogoldtreated rats with CIA compared with their control counterparts. In addition, the histologic score (of synovial hyperplasia, cartilage erosion, and leukocyte infiltration), microvessel density, macrophage infiltration, and levels of TNF␣ and IL-1␤ were also significantly reduced in the ankle joints of nanogold-treated rats.Conclusion. Our results are the first to demonstrate that intraarticular administration of nanogold ameliorates the clinical course of CIA in rats. Nanogold exerted antiangiogenic activities and subsequently reduced macrophage infiltration and inflammation, which resulted in attenuation of arthritis. These results demonstrate proof of principle for the use of nanogold as a novel therapeutic agent for the treatment of

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We have previously reported that polysaccharides extracted from fruiting bodies or cultured mycelia of Antrodia camphorata exhibit an anti-hepatitis B virus effect. In this study, we intended to elucidate the anti-inflammatory potency of six mycelial extracts, namely PDBext, CK-ext, CM-ext, CO-ext, CC-ext, and CKO-ext, isolated from mycelia of A. camphorata cultured with six different media including potato dextrose broth (PDB) and five water-soluble fractions from the wood of different Cinnamomum species, i.e. C. kanehirae (CK), C. micranthum (CM), C. osmophloeum (CO), C. camphora (CC), and C. kotoense (CKO), against reactive oxygen species (ROS) production induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) or phorbol 12-myristate 13-acetate (PMA) in peripheral human neutrophils (PMN) or mononuclear cells (MNC). ROS produced by PMN or MNC act as inflammatory mediators and also signal immune responses. Pretreatment with these mycelial extracts (1^50 Wg ml 31 ) concentration-dependently diminished fMLP-or PMAinduced ROS production in PMN or MNC, as measured by lucigenin-amplified chemiluminescence, with 50% inhibition concentrations (IC 50 ) ranging from 2 to 20 Wg ml 31 . Among these extracts evaluated, CM-ext, CO-ext, or CKO-ext exhibited higher potency than the others. Using high performance liquid chromatography, we identified two lanostane-type compounds, i.e. dehydrosulfurenic acid and 15K-acetyl-dehydrosulfurenic acid, which could be involved in the anti-inflammatory actions of these extracts. The anti-inflammatory actions of these extracts were not due to cytotoxic effects. In summary, these data suggest that extracts from cultured mycelia of A. camphorata display anti-inflammatory effects by inhibiting ROS production in human leukocytes at a pharmacologically applicable concentration. The biological activities of these extracts were further promoted when the culture medium was replaced with water-soluble fractions isolated from the wood of CM, CO or CKO.