Enhanced anti-tumor activity of trichosanthin after combination with a human-derived cell-penetrating peptide, and a possible mechanism of activity.

Trichosanthin (TCS), a type I ribosome-inactivating protein (RIP-I) and renowned Chinese traditional medicine, displays a broad spectrum of biological and pharmacological properties. Particularly, its anti-tumor activity has received a great deal of attention. However, the cellular mechanism for TCS uptake varies with different tumor cell lines, leading to discrepancies in its reported ability to penetrate cells. In this study, HBD, a human derived cell-penetrating peptide (CPP), was used to improve the delivery of TCS into several types of tumor cells, including HeLa cells. Recombinant TCS (rTCS) with or without the fused HBD peptide was expressed in Escherichia coli cells and successfully purified by Ni-NTA affinity chromatography. The cellular uptake efficiency of FITC-labelled-rTCS-HBD was observed in HeLa cells and compared with the uptake efficiency of non-HBD conjugated rTCS under the same conditions using laser confocal microscopy. Moreover, the IC50 value of rTCS-HBD in the tested tumor cells was much lower than that of rTCS, indicating that HBD could efficiently deliver the rTCS into tumor cells. When compared with rTCS, rTCS-HBD induced higher rates of apoptosis in HeLa cells as analyzed by flow cytometry. Furthermore, the apoptotic events observed in HeLa cells incubated with HBD-fused rTCS included activation of Caspase-9, decrease in the Bcl-2/Bax ratio, and cleavage of PARP. These results strongly suggest the participation of mitochondria in apoptosis. This report illustrates one possible method for achieving the efficient transport of TCS into cells using a CPP as a vector, and increases the likelihood that TCS can be used in the clinic.

Adsorption removal of phosphate from aqueous solution by active red mud.

Red mud is the waste of alumina industry and has high TiO2 and Fe2O3 content which are active components for the adsorption of anion pollutants. In this study, the uptake of phosphate by red mud activated by heat treatment and acid-heat treatment was investigated. The factors influencing the adsorption were also investigated. The result showed that the red mud sample treated using acid-heat method at 80 degrees C with 0.25 mol/L HCl for 2 h achieved the highest phosphate removal. For the heat-activated red mud, the sample heated at 700 degrees C for 2 h preformed better than the other heat treatment. Phosphate removal by the activated red mud was significantly pH dependent, and pH 7 was the optimal pH for phosphate removal. The adsorption fits Langmuir isotherm model well and the maximum adsorption capacities of the acid-heat activated red mud and the heat activated samples were 202.9 mgP/g and 155.2 mgP/g, respectively.