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1.
Microbiology ; (12)2008.
Article in Chinese | WPRIM | ID: wpr-686074

ABSTRACT

The method for analysis and determination the cleavage of soybean sterol, in which the soybean sterol was degraded and the products androst-1,4-diene-,17-dione (ADD) and androst-4-ene-3,17-dion (AD) were developed by Liquid Chromatography-mass Spectrometry. The HPLC conditions adopted were: a All- tima ODS-2 column (250 mm?4.6 mm, 5 ?m), a mobile phase consisted of menthanol-water (70:30), a flow rate of 1.0 mL/min, a room column temperature. and the detective wavelength was 244 nm.The ZMD Micromass electrospray ionization (ESI)-mass spectrometer was employed. In such conditions the corre- sponding HPLC chromatogram and MS spectrum were obtained. The method has a linear ranger of 0.01 mg/mL ~ 0.09 mg/mL, R2 =0.9999, the recoveries of ADD and AD were 102.6% and 105.90%, the RSD of ADD and AD were 3.02%, 3.5% and 3.08%, 3.24%. This method showed high sensitivity, accuracyand easy to perform. It is suitable to analysis the process cleavage of soybean sterol as well as quality control of product.

2.
Chinese Journal of Traumatology ; (6): 195-199, 2007.
Article in English | WPRIM | ID: wpr-236781

ABSTRACT

<p><b>OBJECTIVE</b>To clone and identify one novel regeneration related gene H(3) (CA854305) from the differential expression genes library we had set up before.</p><p><b>METHODS</b>Use the method of Northern blot to detect the different expressions of the novel gene under different situations, employ the technique of in silico cloning to scan the span of the novel gene, and analyze their sequences. Also we used reverse transcription PCR to validate the largest open reading frame.</p><p><b>RESULTS</b>Northern blotting results of H(3) (CA854305) showed that the transplanted group had more efficient and extensive expression than untreated and uninjured groups 5 days after spinal cord injury, while the untreated group had more extensive expression than uninjured group. It implied that H(3) might have some relationship with nerve regeneration after spinal cord injury. From the results of in silico cloning we got a longest contig of 1635 bp and an largest open reading frame of 542 bp from 49 to 591 bp correspondent with the Cozak rules. Reverse transcription PCR validated the largest open reading frame sequence primarily.</p><p><b>CONCLUSIONS</b>We got the sequence of novel gene H(3) which might be one of the regenerationjrelated genes. Key words:Gene library; Genes; Nerve regeneration; Spinal cord injuries.</p>


Subject(s)
Animals , Rats , Base Sequence , Blotting, Northern , DNA, Complementary , Genetics , Gene Library , Molecular Sequence Data , Nerve Regeneration , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Spinal Cord Injuries , Genetics
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