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Zhang D.,Sichuan University | Cao M.,Key Laboratory of Sichuan Academy of Medical science | Nie Y.,Sichuan University | Jiang N.,Key Laboratory of Sichuan Academy of Medical science | And 6 more authors.
Advanced Science Letters | Year: 2012

Biot2-L is a novel mouse cancer-testis (CT) antigen gene that was first identified with cross-reactive serological analysis of recombinant cDNA expression libraries technique in our laboratory. To express and purify the CT antigen Bio2-L for further investigation as tumor vaccine, the full-length of Biot2-L gene was cloned and the recombinant expression vector pET28a(+)-Biot2-L was constructed. The full-length gene was confirmed by restriction enzyme digestion and sequence analysis. After optimization, E. coli BL21 (DE3) bearing the recombinant plasmid was induced with 0.8 mM IPTG for 4 h at 37 °C, and the recombinant BIOT2-L was produced at the level of about 28% of total bacterial proteins. Target protein was purified by Ni 2+ affinity chromatography and characterized by western blot with anti-His antibody. The purified recombinant protein was further confirmed by ESI-Q-TOF-MS. After protein purification, Biot2-L was used to immunize adult rabbits and its antibodies were generated. The titer was 6.4×10 5. This study provided a valuable foundation for further researches on the novel gene, Biot2-L. © 2012 American Scientific Publishers. All rights reserved.

Liu S.,Leshan Teachers College | Cao M.,Key Laboratory of Sichuan Academy of Medical science | Li D.,Academy of Sichuan Grass Science | Zhang J.,Sichuan University | And 7 more authors.
Natural Product Research | Year: 2011

Sinojackia sarcocarpa L.Q. Luo has high aesthetic value, which is clinically used to treat diseases such as the variations of arthritis, thromboangiitis obliterans, angina pectoris as well as many other diseases. Pentacyclic triterpenoids are the main pharmacological effective compounds. High-speed counter-current chromatography method was performed on a Midi-DE centrifuge at 25°C. The solvent system was n-hexane- ethylacetate-methanol- water (10 : 5 : 3 : 1, v/v). Peak fractions were collected according to the elution profile for subsequent high-performance liquid chromatography analysis. The structure identification was performed by ultraviolet, infrared, mass spectrometry, 1H-NMR and 13C-NMR. Compound 2α,3α,19β,23β-tetrahydroxyurs-12-en-28-oic acid and 2α,3α,23β-trihydroxyurs-12-en-28-oic acid were purified from the plant of Styracaceaen genius for the first time, whose purities were 96.57% and 97.33%, respectively. Compared with the same dose of oral 5-fluorouracil with 57.6% inhibition rate, the S 180 tumour inhibition rates of 20mg kg -1 d -1 two compounds were 59.5% and 48.9%, respectively. © 2011 Taylor & Francis.

Ding X.,Sichuan University | Tang J.,Sichuan University | Cao M.,Key Laboratory of Sichuan Academy of Medical science | Guo C.-X.,Sichuan University | And 8 more authors.
International Journal of Biological Macromolecules | Year: 2010

In this study, structural features of Tricholoma matsutake polysaccharide (TMP-A) were investigated by a combination of infrared (IR) spectra, gas chromatography-mass spectrometry (GC-MS), nuclear magnetic resonance (NMR) spectroscopy. The results indicated that TMP-A had a backbone of 1,4-β-d-glucopyranose residue which branches at O-6 based on the experimental results. The branches were mainly composed of an (1→3)-α-d-galactopyranose residue, and terminated with α-d-xylopyranose residue. The antioxidant activity of TMP-A was evaluated with several biochemical methods, including DPPH- radical scavenging, hydrogen peroxide scavenging, superoxide anion radical scavenging. The results indicated that TMP-A showed strong antioxidant. In the in vitro antioxidant assay by MTT method, TMP-A could attenuate PC12 cell damage significantly caused by hydrogen peroxide. © 2010 Elsevier B.V.

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