ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of KCa3.1 channel inhibitor TRAM-34 on the proliferation and invasion of leukemia cell line HL-60.</p><p><b>METHODS</b>HL-60 cells at logarithmic growth phase exposed to TRAM-34 at the final concentration of 25, 50, 75 and 100 nmol/L were used as experimental group. The HL-60 cells of control group was cultured in 10% fetal bovine serum-RPMI 1640. The proliferation inhibition rate of TRAM-34 on HL-60 cells was detected by adding MTT solution after 24, 48 and 72 h culture. The cell apoptotic rate and cell cycle distribution of HL-60 cells treated with TRAM-34 were evaluated by flow cytometry with Annexin V-FITC/propidium iodide(PI) double staining or PI single staining. The number of transmembrane cells was detected by Transwell at 24 and 48 h after treatment with TRAM-34. The effect of TRAM-34 on CDK6, P53 and MMP-2 mRNA level was detected by real-time quantitative PCR.</p><p><b>RESULTS</b>Compared with the control group (0 nmol/L), the inhibition rate, apoptosis rate, G/Gphase cell proportion and P53 mRNA level all increased, but the percentages of cells in S phase, cell number penetrating the membrane and mRNA levels of CDK6 and MMP-2 in the TRAM-34-treated group decreased (P<0.05) except for 24 h proliferation rate of TRAM-34 at low concentration (25 nmol/L). The effect of TRAM-34 on the above indices was enhanced with the increase of concentration and prolongation of time, and the differences were statistically significant (P<0.05).</p><p><b>CONCLUSION</b>TRAM-34 can inhibit the proliferation and invasion of HL-60 cells, and can induce cell apoptosis and G/Garrest. The time and concentration of TRAM-34 have effect on the malignant behavior of HL-60 cells.</p>
ABSTRACT
<p><b>OBJECTIVES</b>To study the effect of cryopreservative period on the cryosurvival of human spermatozoa and find out the optimal recovery time of cryopreservation.</p><p><b>METHODS</b>Eighty-eight semen samples were collected from normal donors and divided randomly into 5 groups according to the period of cryopreservative storage (1 d, 7 d, 30 d, 180 d, 300 d) in liquid nitrogen after being frozen by the programized, three-step freezing method. Fresh and frozen-thawed semen were examined by the routine analysis of semen and then the sperm recovery rate were calculated.</p><p><b>RESULTS</b>There were no significant differences in sperm recovery rate between group I and the others (P > 0.05). The period of cryopreservative storage in liquid nitrogen had no correlation with the cryosurvival of human spermatozoa (r = 0.05, P > 0.05).</p><p><b>CONCLUSIONS</b>It was indicated that freezing-thawing after 24 h would be helpful to the screening of semen donors in batches for donor insemination of human sperm bank.</p>