[Expression of musashi-2 gene in leukemia stem cells from acute myeloid leukemia patients].

This study was aimed to detect the expression of Musashi-2 (Msi2) in acute myeloid leukemia (AML) and investigate the relationship between Msi2 and other clinical parameters, especially CD34. A total RNA was extracted from bone marrow of newly diagnosed AML patietns. The Msi2 mRNA expression in newly diagnosed AML patients was detected with real-time fluorescence quantitative RT-PCR. The expression level of CD34 in above-menthioned patients was detected by flow cytometry (FCM). The relationship between the expression of Msi2 mRNA and clinical outcome in AML patients was analysed. The results showed that (1)the expression of Msi2 mRNA in newly diagnosed AML patients was much higher than that in healthy volunteers (P < 0.05) , especially in M1, M4 and M5 patients; (2)the expression level of Msi2 did not correlate with age, sex, white blood cell count of peripheral blood, AML1/ETO and PML/RARa fusion gene (P > 0.05); (3) Msi2 expression level in patients with CD34(+) cells was significantly higher than that in patients with CD34(-) cells (P < 0.05). It is concluded that the Msi2 mRNA expresses in leukamia stem cells, the high expression of Msi2 mRNA has been found in newly diagnosed AML patients, especially in M1, M4 and M5 patients, the high expression also has been observed in patients with CD34(+).

[Protective effects of inhibition of tissue nitric oxide in the initial stage against tardive injury of testicular spermatogenic function: experiment with rats].

OBJECTIVE: To study the protective effects of inhibition of tissue nitric oxide in the initial stage against the tardive injury of contralateral testicular spermatogenic function after unilateral testicular torsion. METHODS: 56 prepubertal Sprague-Dawley rats were randomly divided into 4 equal groups: placebo group undergoing rotation of left testis 720 degrees clockwise for 4 h, fixing thereof to the scrotum, and then intravenous injection of normal saline; cyclosporine group, undergoing intraperitoneal injection of cyclosporine once daily for 1 month after the testicular rotation and fixation; NG-methyl L-arginine (L-NMMA) group, undergoing intravenous injection of L-NMMA 30 mg/kg 30 min before the testicular rotation; and sham-operation group, undergoing isolation and suture of testis only. The right (untwisted) testes were removed from 7 rats from each group 1 week and 2 months after surgery. The malondialdehyde (MDA) level and nitrous oxide (NO)/nitrogen oxide synthase (NOS) content were evaluated. Histological examination was conducted. The mean seminiferous tubule diameter (MSTD) was assessed. The level of MHC peptide-tetramer complex was determined by flow cytometry. RESULTS: The levels of MDA, NO, NOS, and MHC peptide-tetramer complex of the L-NMMA, placebo, and cyclosporine groups one week after surgery were all significantly higher than those of the sham operation group (all P < 0.05). The levels of MDA, NO, NOS, and MHC peptide-tetramer complex of the L-NMMA group were all significantly lower then those of the placebo group (all P < 0.05). The pathological damage of the contralateral testis in early and late stages in the L-NMMA group was lighter than in the placebo group. CONCLUSION: By inhibiting tissue NO production in focal organizations during the early period, L-NMMA reduces the damages inthe testis contralateral to the testis undergoing unilateral torsion.

[Effects of matrine on the apoptosis and expression of adhesion molecule in multiple myeloma RMPI8226 cells].

To investigate the effects of matrine on apoptosis and expression of adhesion molecules in human multiple myeloma cell line RPMI8226 cells, RPMI8226 cells were incubated with indicated concentrations of matrine. The growth of RPMI8226 cells was observed by CCK-8 colorimetric assay and apoptosis was detected by flow cytometry using Annexin V-FITC/PI staining. The cell cycles were analyzed by PI staining. Flow cytometry using Annexin V-FITC/PI staining was used to detect the expression of cell adhesion molecules, including CD44, CD44v6, CD54 and CD106. The results showed that RPMI8226 cell viability in presence of matrine decreased markedly in a dose- and time-dependent manners. The apoptosis could be induced by matrine and its level increased following the augmentation of the drug concentration. After treated by matrine for 48 hours, a concentration-dependent increase of cells in G(0)/G(1) phase and a decrease in S phase could be detected, but no obvious change of cell count was found in G(2)/M phase. Treatment of RPMI8226 cells with matrine for 48 hours resulted in decrease of expression levels of CD44 and CD54, while expressions of CD44v6 and CD106 had no significant change. It is concluded that matrine induces in vitro apoptosis, suppresses proliferation in multiple myeloma cells and depresses expression of some adhesion molecules.

Inhibition of mitochondria responsible for the anti-apoptotic effects of melatonin during ischemia-reperfusion.

OBJECTIVE: To investigate a possible mechanism responsible for anti-apoptotic effects of melatonin and provide theoretical evidences for clinical therapy. METHODS: Ischemia-reperfusion mediated neuronal cell injury model was constructed in cerebellar granule neurons (CGNs) by deprivation of glucose, serum and oxygen in media. After ischemia, melatonin was added to the test groups to reach differential concentration during reperfusion. DNA fragmentation, mitochondrial transmembrane potential, mitochondrial cytochrome c release and caspase-3 activity were observed after subjecting cerebellar granule neurons to oxygen-glucose deprivation (OGD). RESULTS: The results showed that OGD induced typical cell apoptosis change, DNA ladder and apoptosis-related alterations in mitochondrial functions including depression of mitochondrial transmembrane potential (its maximal protection ratio was 73.26%) and release of cytochrome c (its maximal inhibition ratio was 42.52%) and the subsequent activation of caspase-3 (its maximal protection ratio was 59.32%) in cytoplasm. Melatonin reduced DNA damage and inhibited release of mitochondrial cytochrome c and activation of caspase-3. Melatonin can strongly prevent the OGD-induced loss of the mitochondria membrane potential. CONCLUSION: Our findings suggested that the direct inhibition of mitochondrial pathway might essentially contribute to its anti-apoptotic effects in neuronal ischemia-reperfusion.