RESUMO
Under low temperature conditions, the hapten carboxyl-norketamine was synthesized by reacting norketamine and succinaldehyde acid.Identification result using electrospray ionization mass showed the hap ten was successfully synthesized.The artificial antigen confirmed by infrared spectroscopy was developed by conjugating hapten to carrier proteins with carbodiimide(EDC) method.Matrix-assisted laser desorption ioni zation time of flight mass spectrometry(MALDI-TOF-MS) showed that the ratio of hapten to BSA was 11:1.The antibody with high titer(5.12 × 10~4) was produced after immuning to rabbits.
RESUMO
Objective To investigate the correlation between aberrant methylation status of ras association domain family 1A (RASSF1A) , transcriptional levels of DNA methyltransferases (DNMTs) and hepatitis B virus ( HBV) infection in patients with hepatocellular carcinoma ( HCC). Methods HCC samples in 61 cases were collected. Aberrant methylation status of RASSF1A was detected using methylation specific PCR (MSP). Transcriptional levels of DNMT1, DNMT3 A and DNMT3B was measured using semi-quantitative reverse transcriptase-PCR(RT-PCR). Results RASSF1A gene with abnormal methylation in initiation zone was found in 45 cases with HCC among 61 patients (73.8%). Further analysis revealed RASSF1A methylated in 32 of 47 HBV infected cases and in 13 of 14 uninfected cases. However, there was no significant association between methylation status of RASSF1A and HBV infection (x2 =2.260, P = 0. 133). Compared with normal control, DNMTs was up-regulated in all HCC samples, HCC cell lines and HBV infected group. Analysis within each group indicated that DNMT3A and DNMT3B levels of HCC increased in MSP positive cases ( t = 3. 494, P