AIM:To explore the effect of dasatinib on the viability, migration, cell cycle and apoptosis of human bone marrow mesenchymal stem cells (hBMSCs), as well as the underlying signal pathway to evaluate the influence of dasatinib on hematopoietic microenvironment clinically.METHODS:The cell viability was measured by CCK-8 assay.The migration ability was detected by wound healing assay.The cell cycle and apoptosis were analyzed by flow cytometry.Acridine orange/ethidium bromide staining was also used to detected apoptosis.The secretion of transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were measured by ELISA.The protein levels of cleaved caspase-3, protein kinase B (Akt) and phosphorylated Akt were determined by Western blot.RESULTS:Compared with control group, dasatinib at 1~10 nmol/L suppressed the viability and migration ability of hBMSCs, and dasatinib at concentration of 7 nmol/L was adopted in the following assays.Dasatinib promoted apoptosis, and blocked the cell cycle in G1 phase.In addition, the secretion of TGF-β1 and TNF-α was increased markedly.The protein levels of cleaved caspase-3 was increased, but the protein levels of Akt and phosphorylated Akt were decreased.CONCLUSION:Dasatinib inhibits the viability and migration ability of hBMSCs in a dose-dependent manner, promotes the secretion TGF-β1 and TNF-α, and induces cell cycle arrest and apoptosis.Dasatinib might regulate the biological behaviors of hBMSCs observed above by modulating the expression and phosphorylation of Akt.

AIM:To explore the protective effects of salidroside on endothelial progenitor cells (EPCs) dam-aged by radiation and its mechanisms.METHODS:EPCs from normal peripheral blood were cultured in fibronectin-coated flasks with endothelial progenitor medium.The effects of salidroside on the viability, migration, adhesion and apoptosis of radiation-damaged EPCs were detected.The viability, apoptosis and migration of the cells were assayed by CCK-8 assay, flow cytometry and Transwell chamber experiment, respectively.The cell adhesion assay was performed by re-plating the cells on fibronectin-coated dishes, and then the adherent cells were counted.The expression of Akt protein in the cells was assessed by Western blotting.RESULTS:Salidroside improved the viability, and migratory and adhesive capacities of the EPCs, and decreased the apoptosis after radiation.Salidroside also increased the protein level of phosphorylated Akt.How-ever, the effects of salidroside on radiation-damaged EPCs were inhibited by phosphatidylinositol 3-kinase inhibitor LY294002.CONCLUSION: Salidroside protects EPCs from radiation damages and its mechanism is associated with en-hancing phosphatidylinositol 3-kinase/Akt signaling pathway.

AIM:Toinvestigatetheeffectofnicotinicacidamide(NAA)ontheinfusiondamageofhuman umbilical cord mesenchymal stem cells ( hUC-MSCs) under the condition of instant blood-mediated inflammatory reaction ( IBMIR) .METHODS:Normal peripheral blood without anticoagulant at volume of 2.7 mL was mixed with 0.3 mL phys-iological saline (as blank group), CFSE labeled hUC-MSCs (1 ×106 cells in 0.3 mL as MSC group) and CFSE labeled hUC-MSCs (1 ×106 cells in 0.3 mL) preprocessed with NAA at concentration of 10 mmol/L for 24 h ( as MSC+NAA group) , respectively.The mixture was immediately injected into the improved Chandler Loop model, placed in 37℃water bath, and then started the peristaltic pump at the speed of 20 mL/min for 1 h.The number of CFSE labeled hUC-MSCs, platelets, white blood cells were counted and the concentration of complement C3a was measured before and after cycling, respectively.RESULTS: After 1 h circulation, the platelet dissipation rate were ( 29.96 ±10.88 )% in blank group, (77.76 ±19.29)% in MSC group all and (50.13 ±18.10)% in MSC +NAA group; and the leukocyte counts were (37.82 ±13.81)%in blank group, (64.57 ±17.08)% in MSC group and (41.52 ±17.26)% in MSC+NAA group. Compared with blank group, the differences of the dissipation rates in MSC group and MSC+NAA group all had statistical significance.The hUC-MSCs relative survival rate in MSC+NAA group was higher than that in MSC group.C3a concentra-tions in blank group, MSC group and MSC+NAA group were (206.27 ±58.10), (230.47 ±39.61) and (208.37 ± 40.66) μg/L, respectively.CONCLUSION:Co-circulating the mixture of hUC-MSCs with normal peripheral blood with-out anticoagulant in the improved Chandler Loop for 1 h depletes a large number of hUC-MSCs and blood components, and increases C3a, suggesting that this model can induce IBMIR.NAA has a protective effect on the hUC-MSCs in the infusion damage by inhibiting IBMIR, reducing the wastage of the blood components and enhancing the survival rate of the hUC-MSCs.

[ABSTRACT]AIM:ToinvestigatetheeffectofmiR-155-specificsiRNAaloneorincombinationwithcytosinear-abinoside (Ara-C) on the growth and apoptosis of Burkitt lymphoma Raji cells .METHODS: miR-155-specific siRNA and/or Ara-C were used to treat the cells .Quantitative real-time polymerase chain reaction was used to detect the expres-sion of miR-155.The growth of the cells was analyzed by CKK-8 assay.The cell apoptosis was determined by flow cytome-try.RESULTS:The miR-155 expression level of the cells transfected with miR-155 siRNA was significantly lower than that in the 2 control groups .Ara-C or miR-155 siRNA alone inhibited the growth of Raji cells in a dose-depend manner . miR-155 siRNA combined with Ara-C produced more inhibition of cell proliferation (P<0.05).After treatment for 48 h, the apoptotic rate of Raji cells in miR-155 siRNA+Ara-C group [(38.4 ±1.4)%] was higher than that in Ara-C group [(16.5 ±0.3)%] and miR-155 siRNA group [(14.6 ±0.3)%], with statistically significant difference (P<0.05). The expression of caspase-3 in Ara-C+miR-155 siRNA group was increased significantly as compared with Ara-C group and miR-155 siRNA group.CONCLUSION:miR-155-specific siRNA enhances the chemosensitivity of Raji cells to Ara-C by inducing apoptosis through the caspase-3 pathway .

Objective To investigate the potential and underlying molecular mechanism of salidroside in ameliorating radiation-induced bone marrow adipogenesis and stimulating hematopoiesis.Methods The female BALB/c mice aged 6-7 weeks were randomly divided into normal control group,radiation group and salidroside group.The radiation group and salidroside group were irradiated with 6.0 Gy of 60Co γ-rays.The salidroside group was intraperitoneally injected with 30 mg· kg-1 · d-1 salidroside at 12 h and then every day until 8th d after radiation.The normal control group and radiation group were treated with equal volume of saline as control of salidroside.At 14 d after radiation,the mice weight,peripheral blood count,femur bone marrow histology,and the proportion of adipocyte area were measured,and the expressions of PPAR-γ and FABP4 were detected by q-PCR.Results After irradiation,the numbers of white blood cells,hemoglobin and platelet in peripheral blood were reduced obviously,and the percentage of adipocyte area was increased significantly.Compared with mice in the radiation group,salidroside inhibited adipogenesis and reduced the proportion of adipocyte area (t =13.31,P ＜ 0.05) by reducing the expressions of PPAR-γ and FABP4 (t =8.64,13.19,P ＜ 0.05).The number of white blood cells was partly recovered at 7 d after irradiation (t =5.80,P ＜ 0.05).Both white blood cells and hemoglobinin in peripheral blood of the salidroside group were higher than those in the radiation group at 14 d after irradiation.Conclusions Salidroside could inhibit radiation-induced bone marrow adipogenesis and regulate bone marrow microenvironment,thereby promotes hematopoietic recovery in mice after radiation injury.

AIM: To investigate whether salidroside has influence on the activities of endothelial progenitor cells (EPCs) and its mechanism.METHODS:Mononuclear cells from normal human peripheral blood were cultured in fi-bronectin coated flasks in endothelial progenitor medium .After 7 d, EPCs were characterized as adherent cells with acLDL-DiI uptaking and lectin binding by direct fluorescent staining .The proliferation and migration of EPCs were analyzed by MTT assay and Transwell chamber assay , respectively.The EPCs adhesion assay was performed by re-plating the cells on fibronectin-coated dishes , and then adherent cells were counted .NO and Akt protein were also detected .RESULTS:Sali-droside promoted EPCs proliferative , migratory and adhesive capacities in a concentration dependent manner .Salidroside also increased NO secretion , and the level of phosphorylated Akt protein .However , the effects of salidroside on EPCs were inhibited by phosphoinositide 3-kinase inhibitor LY294002.CONCLUSION:Salidroside regulates the activity of EPCs by phosphoinositide 3-kinase/Akt signaling pathway .

Objective To investigate the protective effect of 1,25-(OH) 2D3 on radiation-induced bone marrow microenvironment injury and to explore the related molecular mechanism.Methods Sixty 7-week old male BALB/c mice were randomly divided into control group without any treatment; radiation group exposed to 6.0 Gy 60Co γ-rays with DMSO,and 1,25-(OH)2 D3 + radiation group treated with 1,25-(OH)2D32.5 μg/kg dissolved in DMSO each day and 6 Gy of γ-rays.The body weight and peripheral white blood cells,femur bone marrow histology,and the proportion of adipocyte area were measured.The expression of peroxisome proliferator-activated receptor-gamma (PPARγ) was detected immunohistochemistrically at 8 d after irradiation.Results After irradiation,the number of white blood cells and the body weight decreased obviously,and the percentage of adipocyte area was increased significantly.Compared with radiation group,1,25-(OH)2D3 reduced the decrease rate of body weight (t =-2.23,-2.34,P ＜ 0.05),partly recovered the number of white blood cells at 4 or 8 d after irradiation(t =-4.99,-4.46,P ＜ 0.05),and reduced the proportion of adipocyte area (t =-3.75,-2.10,P ＜ 0.05).With immunohistochemistrical assay,it was found that 1,25-(OH) 2D3 inhibited adipogenesis by reducing the expression of PPARγ.Conclusions 1,25-(OH) 2 D3 decreases radiationinduced adipogenesis and hence protects the bone marrow microenvironment from radiation damage.

To determine the role of placenta cells allogeneic graft in mammalian longevity, the 15-month-old female BALB/c mice (n = 50) were divided into Control group (A), Short-term transplanted group (B) and Long-term transplanted many times group (C). Their placentae (at 18 days of gestation) were taken out and ground with 50-eye cell grit, and the cells were intraperitoneally injected into the mice of B group and C group three times at intervals of 10 days; then the cells were transplanted into the mice of C group many times till the time of death. The mice were evaluated by use of ultrasound-cardiogram; autopsy; score of cardia, spleen, skin, lung, kidney; histopathology; serum total superoxide dismutase activity, serum maleic dialdehyde content, and serum glutathione peroxidase activity. The long-term surviving stem cells were found to be located in many organ tissues of B and C groups' mice with in situ Y chromosomal hybridization dyeing. Median life span of B group mice was 1.7 times that of A group's after transplantation, but there was no statistically significant difference between B group and C group. Three months after transplantation, in B group, the pathological developments of significant skin, cardia, lung, and kidney were delayed; the retrogradation of heart function was attenuated; the data on heart mass index (mass of heart/mass of body), left ventricular mass and serum Maleic Dialdehyde content, and on spleen mass index (mass of spleen/mass of body), left ventricular diastolic volume, serum Total Superoxide Dismutase activity and serum Glutathione Peroxidase activity, were all in a direction favourable to B group (P < 0.05). These results were in line with the hypothesis, i. e. longevity can be enhanced to some extent by transplantation of placenta cells.
Aging ; physiology ; Animals ; Cell Transplantation ; methods ; Female ; Mice ; Mice, Inbred BALB C ; Placenta ; cytology ; Pregnancy ; Random Allocation

Lipid second messengers,particularly phosphoinositides,play a pivotal role in several cell signaling networks.Specific inositol lipids such as PtdIns(3,4)P_(2) and PtdIns(3,4,5)P_(3) which are generated by phosphoinositide 3-kinases(PI3Ks) are specific activators to the serine/threonine protein kinase Akt that have been implicated in a plethora of cell functions.Studies show that Akt stimulates cell proliferation and suppresses apoptosis,and is implicated in cancer progression.Many evidences have highlighted the presence of an autonomous nuclear inositol lipid metabolism,and suggest that lipid molecules are important components of signaling pathways in the nucleus.

OBJECTIVETo study the features of vascular smooth muscle cell (VSMC) proliferation induced by endothelin-1 (ET-1).

METHODSVSMCs of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats were cultured and treated with ET-1. Basic fibroblast growth factor (bFGF) gene expression was measured using both Northern blot and an enzyme-linked immunoassay.

RESULTSET-1 resulted in an increase in bFGF transcripts at 8 - 24 h; bFGF levels were significantly higher in VSMCs treated with ET-1 than in those not treated. However, VSMCs growth responses in SHR and WKY were different. Smooth muscle cells of SHR were hyper-responsive to ET-1. Maximal bFGF mRNA levels were elevated 3.5-fold at 4 h of stimulation in WKY and 8-fold at 8h in SHR4. Moreover, the proliferation of VSMCs induced by ET-1 was inhibited by antisense phosphorothioate oligodeoxynucleotides (10 micromol/L AS-bFGF) but not sense bFGF oligomers at the same concentrations, being reduced by 80% in SHR and 40% in WKY vs control, respectively. Furthermore, the effect of AS-bFGF oligomers on SHR SMC proliferation is significantly greater than on WKY SMC proliferation.

CONCLUSIONET-1 may be required for exaggerated vascular growth responses in SHR and bFGF may be involved.

Animals ; Cell Division ; drug effects ; Cells, Cultured ; DNA, Antisense ; pharmacology ; Dose-Response Relationship, Drug ; Endothelin-1 ; pharmacology ; Fibroblast Growth Factor 2 ; genetics ; Gene Expression Regulation ; drug effects ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Oligonucleotides ; pharmacology ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Time Factors