Detection of EGFR mutations in pleural effusion of patients with non-small cell lung cancer / 实用医学杂志

Objective To detect the consistency of EGFR mutations in pleural effusion in terminal non-small cell lung cancer(NSCLC)patients by amplification refractory mutation system- polymeric chain reaction (ARMS-PCR).Methods Pleural effusion from 44 cases with advanced NSCLC were collected.DNA was extract-ed from a part of pleural effusion,and EGFR gene18,19,20,and 2 l exons mutations were detected by ARMS-PCR amplification. The left part of pleural effusion was used for the pathological microscopic examination of the embedding and to evaluate the feasibility of pleural effusion direct detection.Part of the case results were compared with tissue or pleural effusion at the same time. Results The mutation rate of EGFR detection in pleural effusion was 54.5%,including 19-del(50.0%)and 21-L858R(50.0%);75.0% of pleural effusion sediment was morpho-logically diagnosed with adenocarcinoma and EGFR mutation rate was 27.3% in pleural effusion with no tumor cell. The consistent rate was 72.7% in 22 cases of pleural effusion and tissue and that was 50.0% in 16 cases of pleural effusion and plasma.The comparison between pleural effusion and organizations and plasma test results was statisti-cally significant(P < 0.05).(P < 0.05).Conclusions EGFR gene mutation rate in pleural effusion is lower than that in tumor tissue specimens in terminal NSCLC patients and tumor tissue is still the best test specimens.

De novo synthesis of glutathione is a prerequisite for curcumin to inhibit hepatic stellate cell (HSC) activation.

On liver injury, quiescent hepatic stellate cells (HSC), the most relevant cell type for hepatic fibrogenesis, become active, characterized by enhanced cell growth and overproduction of extracellular matrix (ECM). Oxidative stress facilitates HSC activation and the pathogenesis of hepatic fibrosis. Glutathione (GSH) is the most important intracellular antioxidant. We previously showed that curcumin, the yellow pigment in curry from turmeric, significantly inhibited HSC activation. The aim of this study is to elucidate the underlying mechanisms. It is hypothesized that curcumin might inhibit HSC activation mainly by its antioxidant capacity. Results from this study demonstrate that curcumin dose and time dependently attenuates oxidative stress in passaged HSC demonstrated by scavenging reactive oxygen species and reducing lipid peroxidation. Curcumin elevates the level of cellular GSH and induces de novo synthesis of GSH in HSC by stimulating the activity and gene expression of glutamate-cysteine ligase (GCL), a key rate-limiting enzyme in GSH synthesis. Depletion of cellular GSH by the inhibition of GCL activity using L-buthionine sulfoximine evidently eliminates the inhibitory effects of curcumin on HSC activation. Taken together, our results demonstrate, for the first time, that the antioxidant property of curcumin mainly results from increasing the level of cellular GSH by inducing the activity and gene expression of GCL in activated HSC in vitro. De novo synthesis of GSH is a prerequisite for curcumin to inhibit HSC activation. These results provide novel insights into the mechanisms of curcumin as an antifibrogenic candidate in the prevention and treatment of hepatic fibrosis.

The antifibrogenic effect of (-)-epigallocatechin gallate results from the induction of de novo synthesis of glutathione in passaged rat hepatic stellate cells.

Hepatic stellate cells (HSC) are the major players during hepatic fibrogenesis. Overproduction of extracellular matrix (ECM) is a characteristic of activated HSC. Transforming growth factor-beta (TGF-beta) is the most potent fibrogenic cytokine while connective tissue growth factor (CTGF) mediates the production of TGF-beta-induced ECM in activated HSC. HSC activation and hepatic fibrogenesis are stimulated by oxidative stress. Glutathione (GSH) is the most important intracellular antioxidant. The aim of this study is to explore the mechanisms of (-)-epigallocatechin-3-gallate (EGCG), the major and most active component in green tea extracts, in the inhibition of ECM gene expression in activated HSC. It is hypothesized that EGCG inhibits ECM gene expression in activated HSC by interrupting TGF-beta signaling through attenuating oxidative stress. It is found that EGCG interrupts TGF-beta signaling in activated HSC by suppressing gene expression of type I and II TGF-beta receptors. EGCG inhibits CTGF gene expression, leading to the reduction in the abundance of ECM, including alphaI(I) procollagen. Exogenous CTGF dose dependently eliminates the antifibrogenic effect. EGCG attenuates oxidative stress in passaged HSC by scavenging reactive oxygen species and reducing lipid peroxidation. De novo synthesis of GSH is a prerequisite for EGCG to interrupt TGF-beta signaling and to reduce the abundance of alphaI(I) procollagen in activated HSC in vitro. Taken together, our results demonstrate that the interruption of TGF-beta signaling by EGCG results in the suppression of gene expression of CTGF and ECM in activated HSC in vitro. In addition, our results, for the first time, demonstrate that the antioxidant property of EGCG derived from de novo synthesis of intracellular GSH plays a critical role in its antifibrogenic effect. These results provide novel insights into the mechanisms of EGCG as an antifibrogenic candidate in the prevention and treatment of liver fibrosis.

Fingerprint differences between total flavonoids and coumarins of Sarcandra glabra extract / 中成药

AIM: To compare the fingerprints difference between total flavonoids and coumarins of Sarcandra glabra extract separated by macroporous adsorption resin. METHODS: To establish HPLC fingerprint chromatogram of Sarcandra glabra extract and to evaluate the constituents. RESULTS: The difference fingerprint was showed in total flavonoid part and coumarin part. CONCLUSION: Drug action part could be separated by macroporous adsorption resin and exosyndromed by Spectrogram fingerprints.