Objective:To investigate the expression of SHIP1 in the patients with acute myeloid leukemia and its effect on the apoptosis of human leukemia cells.Methods:The expression of SHIP1 in the bone marrow of patients with acute myeloid leukemia was detected by Westem blot.U937 cells was transfected with SHIP1 expression vector (pEGFP-SHIP1 group) and empty vector control (pEGFP group) respectively,U937 cells without transfection were used as the control group.Flow cytometry was used to detect the apoptosis of the cells,the expression of SHIP1,Bcl-2,Bax,Akt,p-Akt were detected by western blot.Results:The expression of SHIP1 in the bone marrow of patients with acute myeloid leukemia was significantly lower than that of the normal human bone marrow SHIP 1 (P＜0.01).The SHIP1 and Bax expressions as well as the apoptotic rate ofpEGFP-SHIP1 group were significantly higher than those of the control group(P＜0.01),while the Bcl-2 and p-Akt expressions were significantly lower than those in the control group(P＜0.01).Conclusions:SH-P1 expression was down regulated in the bone marrow of patients with acute myeloid leukemia.SHIP1 could promote the apoptosis of human leukemia cells via Akt signaling pathway.

COP9 Signalosome Complex ; Carcinoma, Hepatocellular ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; chemistry ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Gene Expression Regulation, Neoplastic ; Humans ; Intracellular Signaling Peptides and Proteins ; chemistry ; genetics ; metabolism ; Peptide Hydrolases ; chemistry ; genetics ; metabolism ; Phosphorylation ; Phosphoserine ; metabolism

OBJECTIVETo observe p21-activated kinase 6 (PAK6) expression and its possible role after spinal cord injury (SCI) in adult rat.

METHODSSprague-Dawley rats were subjected to spinal cord injury. To explore the pathological and physiological significance of PAK6, the expression patterns and distribution of PAK6 were observed by Western blot, immunohistochemistry and immunofluorescence.

RESULTSWestern blot analysis showed PAK6 protein level was significantly up-regulated on day 2 and day 4, then reduced and had no up-regulation till day 14. Immunohistochemistry analysis showed that the expression of PAK6 was significantly increased on day 4 compared with the control group. Besides, double immunofluorescence staining showed PAK6 was primarily expressed in the neurons and astrocytes in the control group. While after injury, the expression of PAK6 was increased significantly in the astrocytes and neurons, and the astrocytes were largely proliferated. We also examined the expression of proliferating cell nuclear antigen (PCNA) and found its change was correlated with the expression of PAK6. Importantly, double immunofluorescence staining revealed that cell proliferation evaluated by PCNA appeared in many PAK6-expressing cells on day 4 after injury.

CONCLUSIONThe up-regulation of PAK6 in the injured spinal cord may be associated with glial proliferation.

Animals ; Astrocytes ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; metabolism ; Up-Regulation ; p21-Activated Kinases

Mycena subpiligera, a new taxon in sect. Fragilipedes that can strongly enhance the germination efficiency of Gastrodia elata seeds, was discovered in subtropical areas of China. As revealed by a morphological comparison with related Mycena species as well as maximum likelihood (ML) and Bayesian phylogenetic analyses based on sequences of the internal transcribed spacer (ITS) and the large subunit (LSU) regions of nuclear ribosomal RNA, the new taxon can be distinguished from phenotypically similar and phylogenetically related species. Optimal cultural conditions for M. subpiligera basidiomata are reported, and the germination rate of the new species is compared with that of M. citrinomarginata.

OBJECTIVETo investigate the expression and relationship of p27(kip1) and its related molecules Jab1 and CRM1 during proliferation of lymphoma cells U937.

METHODSU937 cells were treated with serum starvation and release, and the effects of these treatments on the cell growth was tested with cell number counting. The expression and localization of p27(kip1), Jab1 and CRM1 in U937 cells were detected by Western blot, double immunolabelling and laser scanning confocal microscopy.

RESULTSThe growth of U937 cells was blocked by serum starvation. The total protein of p27(kip1) was increased while Ser10-phosphorylated p27(kip1) -related molecules Jab1 and CRM1 were decreased. Meanwhile, the location of p27(kip1) was changed from cytoplasm into nuclei. After serum release, the location of p27(kip1) expression reappeared in the cytoplasm again.

CONCLUSIONDuring the proliferation process of lymphoma U937 cells, Jab1 and CRM1 may influence the location and expression of p27kip1, and may participate in regulation of growth of NHL cells.

COP9 Signalosome Complex ; Cell Culture Techniques ; Cell Nucleus ; metabolism ; Cell Proliferation ; Culture Media, Serum-Free ; pharmacology ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cytoplasm ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Karyopherins ; metabolism ; Peptide Hydrolases ; metabolism ; Receptors, Cytoplasmic and Nuclear ; metabolism ; U937 Cells

OBJECTIVETo investigate effect of tumor necrosis factor-alpha (TNF-alpha) on the Src-suppressed C kinase substrate (SSeCKS) in C6 glioma cells.

METHODSCultured C6 glioma cells were randomly divided into two groups. In time-dependent group, cells were cultured with TNF-alpha (2 ng/mL) for 0 h, 1 h, 3 h, 6 h, 12 or 24 h, respectively; in dose-dependent group, cells were cultured with TNF-alpha (0 ng/mL, 0.02 ng/mL, 0.2 ng/mL, or 2 ng/mL) for 6 h. The expression of SSeCKS was detected by Realtime PCR and Western blot analysis, and immunocytochemistry was used to investigate SSeCKS's subcellular localization.

RESULTSTNF-alpha induced rapid phosphorylations of protein kinase C (PKC) substrates in C6 glioma cells, and upregulated SSeCKS expression in a time and concentration dependent manner. Immunocytochemistry suggested that SSeCKS was localized in the cyroplasm and the leading end of podosomal extensions in control groups, while TNF-alpha induced translocation of SSeCKS perinuclear. This effect could be partly reversed by PKC inhibitor Ro-31-8220.

CONCLUSIONTNF-alpha activates PKC and upregulates SSeCKS expression in C6 glioma cells. These effects are associated with PKC activity, suggesting that SSeCKS plays a role in response to glia activation in PKC mediated pathway.

A Kinase Anchor Proteins ; Animals ; Astrocytes ; metabolism ; Brain Neoplasms ; metabolism ; Cell Cycle Proteins ; metabolism ; Dose-Response Relationship, Drug ; Enzyme Activation ; Gene Expression Regulation ; Glioma ; metabolism ; Immunohistochemistry ; Protein Kinase C ; metabolism ; Protein Transport ; physiology ; Random Allocation ; Rats ; Signal Transduction ; physiology ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; administration & dosage ; physiology

OBJECTIVETo investigate the expression and relationship of p27(kip1) and its nuclear export factor Jab1 during proliferation process of lymphoma cell.

METHODSJurkat and Raji cells were treated with serum starvation and then serum release. The protein and mRNA expression of p27(kip1), Jab1 in the cells were detected by Western blot and RT-PCR respectively. LMB were used for stimulating Jurkat cells during their proliferation process, and then the expression changes of p27(kip1) and Jab1 were detected. An eukaryotic expression plasmid(pcDNA3. 1-myc) containing Jab1 was constructed. Jurkat cell were transfected in vitro with or without pcDNA3. 1-myc-Jab1. Double immunolabelling was used to identify the localization of p27(kip1). Immunoprecipitation was used to detect the combination of p27(kip1) and Jab1.

RESULTSThe growth of Jurkat and Raji cells were blocked by serum starvation. The total protein amount of p27(kip1) increased while that of Jab1 decreased. The reverse changes were happened after serum release, but the mRNA expression of p27(kip1) has no significant change. LMB could inhibit the cell proliferation caused by serum release. The expression of p27(kip1) was up-regulated and Jab1 down-regulated when Jurkat cells were treated with LMB. After pcDNA3. 1-myc-Jab1 infected Jurkat cells for 48 h, the distribution of p27(kip1) was translocated from nucleus into cytoplasma. p27(kip1) and Jab1 could form compound in Jurkat and Raji cells detected by Immunoprecipitation.

CONCLUSIONJab1 may influence the location and expression of p27(kip1) through integrating with p27(kip1), and then participates in regulating the growth of NHL cell through interfering with the function of p27(kip1).

COP9 Signalosome Complex ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Jurkat Cells ; Peptide Hydrolases ; metabolism ; Plasmids ; RNA, Messenger ; metabolism ; Transfection

OBJECTIVETo investigate the inductive factors of effect on Akebia trifoliata and establish culture method for A. trifoliata callus.

METHODTo study the possible effective factors of culture condition by comparing with different explantation, nutrient medium, pH, temperature, illumination, growth substance of plant and its ratio.

RESULTThe inductivity of leaves was the highest about 87.5%, followed with the stem section and leafstalk; The inductivity of nutrient medium such as MS, B5 callus was higher than the ones like H, SH and the White callus amended one; It was found that low-grade Phvalue benefits the growth of callus. The experiment result showed that different pH showed little difference in quality. The best condition of culture was 25 degrees C in temperature.

CONCLUSIONThe best culture condition for callus was the leaves as explantation. The A. trifoliata callus culture's best inductive condition was MS +2, 4-D 4.0 mg x L(-1) + NAA 1.0 mg x L(-1) + KT 1.0 mg x L(-1) (pH 5.8), cultural temperature was 25 degrees C, cultivation was dark.

Culture Media ; chemistry ; pharmacology ; Hydrogen-Ion Concentration ; Magnoliopsida ; drug effects ; growth & development ; Plant Leaves ; drug effects ; growth & development ; Plant Stems ; drug effects ; growth & development ; Plants, Medicinal ; drug effects ; growth & development ; Temperature ; Tissue Culture Techniques ; methods

OBJECTIVETo study the effect of all-trans retinoic acid (ATRA) on U937 cell growth and its mechanism.

METHODSCell cycle was detected by flow cytometry (FCM), expressions of cell cycle associated protein and the p27 related protein were detected by Western blot. The binding of P27 and Skp2 was detected by immunoprecipitation.

RESULTSFCM displayed that ATRA could inhibit the proliferation of U937 cells. At 72 h on 1 micromol/L ATRA treatment, 72% of the cells were arrested at G0/G1 phase. Western blot displayed that ATRA could decrease the expression of cyclin A, up-regulate the expression of p21 and p27, and down-regulate the expression of p27 related proteins Skp2. p27 could bind with Skp2 in U937 cells as detected by immunoprecipitation.

CONCLUSIONATRA may arrest the proliferation of U937 cells through the reduction of Skp2 expression, and finally the induction of the accumulation of p27.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; S-Phase Kinase-Associated Proteins ; metabolism ; Tretinoin ; pharmacology ; U937 Cells

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Staging ; Phosphorylation ; S-Phase Kinase-Associated Proteins ; metabolism