Antiproliferative effect of silencing LSD1 gene on Jurkat cell line and its mechanism / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of silencing LSD1 gene by RNA interference on the proliferation, apoptosis on human lymphocytic leukemia Jurkat cell line and its mechanism.</p><p><b>METHODS</b>The hairpin- like oligonucleotide sequences targeting LSD1 gene was transfected into Jurkat cells by lipofectamine(TM) 2000. The LSD1 mRNA and protein were detected by RQ- PCR and Western blot. Cell growth was determined by MTT. Cell apoptosis was analyzed by flow cytometry. The expression of Bcl-2, Bax, procaspase- 3, and histone H3K4me, H3K4me2, H3K4me3, Act- H3, H3K9me were detected by Western blot.</p><p><b>RESULTS</b>LSD1 mRNA was markedly suppressed by the shRNA targeting LSD1. LSD1 shRNA suppressed the proliferation and induced cells apoptosis of Jurkat cells. The cell apoptotic rate was (41.34±3.58)%, (3.45±1.54)%, (1.76±0.52)% in LSD1 shRNA, Neg-shRNA and Blank respectively, the difference among them was statistically significant (P<0.05). LSD1 shRNA down- regulated the expressions of Bcl- 2 and procaspase- 3, and up- regulated the expression of Bax. The methylation of H3K4me1, me2 and acetylation of Act- H3 improved without change of the methylation of H3K4me3.</p><p><b>CONCLUSIONS</b>Deplete of LSD1 gene maybe through modifying the methylation of histone H3K4 to promote the cell apoptosis and inhibit cell growth in Jurkat cell line.</p>

Therapeutic Effect of Ulinastatin Combined with Xingnaojing Injection for Acute Cerebral Hemorrhage / 广州中医药大学学报

Objective To observe the protective effect to Ulinastatin combined with Xingnaojing Injection (XI) for acute cerebral hemorrhage. Methods One hundred and eight cases of acute cerebral hemorrhage were randomly divided into treatment group ( N=56) and control group ( N=52) . All of the patients were given conventional western medical treatment including relieving cerebral edema, regulating blood pressure, maintaining electrolyte balance, anti-inflammation, preventing epilepsy. Additionally, the control group was given intravenous drip of Ulinstatin, and the treatment group was given intravenous drip of Ulinstatin and XI. The clearance of intracranial hematoma in the two groups was observed on treatment day 3, 7, 14, the absorption of cerebral edema was observed after treatment for one treatment course of 14 days, neurological deficit scores ( NDS) were compared on treatment day 14, 30, and the clinical efficacy was evaluated. Results ( 1) After treatment, hematoma area was reduced in the two groups ( P0.05); on treatment day 7 and 14, hematoma area in the treatment group was less than that of the control group, the difference being significant (P<0.05) . (2) On treatment day 14, the total effective rate for absorption of cerebral edema was 89.3% in the treatment group, and was 65.4% in the control group, the difference being significnat ( P<0.05) . ( 3) After treatment, NDS of the treatment group was less than that of the control group, and the total effective rate on NDS was 89.3% in the treatment, higher than 71.2% in the control group, the difference being significant ( P<0.05) . ( 4) In the treatment group, 3 cases had slight increase of aminotransferase. Eight cases of the control group had increased aminotransferase, and then the aminotransferase level recovered to normal after suspension. In the treatment group, 2 cases were dropped out for receiving emergency operation due to rehaemorrhagia during the treatment, and 3 cases were death for illness deterioration. In the control group, 7 cases gave up the treatment for illness deterioration and then were dead during the follow-up. Conclusion Ulinastatin combined with XI shows certain protective effect in treating acute cerebral hemorrhage.

THE CONSTRUCTION OF TRANSIENT GENE EXPRESSION TRANSFECTION SYSTEM IN MCF-7 CELLS / 解剖学报

Objective To establish an efficient, simple, low cytotoxicity and cheap transfaction system. Methods We have used the cationic polymer polyethylenimine(PEI) to study transient transfection in MCF-7 cells by testing different conditions, including cell concentrations, DNA concentrations, the ratio of PEI nitrogen to DNA phosphate(PEI-N∶DNA-P) and the time of cells grown in serum-free culture together with PEI-DNA complex. Results The optimized cell concentrations were 2?10 5 cells seeded per well in 24-well dishes 18-24?h before transfection. The DNA concentrations and ratio of PEI-N∶DNA-P are very important for optimal transfection and they affect each other. For 1??g DNA per well, the optimal PEI-N∶DNA-P is about 33∶1, however, as for 4??g DNA, it is 9∶1. The best time of cells grown in serum-free culture together with PEI-DNA complex is about 5-7?h.Conclusion With optimized conditions, we can establish an efficient, simple, low cytotoxicity and cheap transfection system by using PEI.