OBJECTIVETo study the effect of emodin on the differentiation, maturation and function of human dendritic cells (DC) in vitro.

METHODSCells isolated from human peripheral blood mononuclear cells (PBMCs) were induced to dendritic cells (DC) with recombinant interleukin-4 and recombinant granulocyte-macrophage colony stimulating factor (GM-CSF). Lipopolysaccharide (LPS) and different concentrations of emodin were added respectively in the cultured cells on the 5th and the 7th to obtain mature or immature DCs. The phenotype of DCs ( HLA-DR, CD80, CD86, CD83, CD14, CD11c) and the secretion of interleukin-12 (IL-12) were analyzed by flow cytometry, and the immune-stimulating function of DCs was evaluated by co-culture of DCs and self-T-lymphocytes.

RESULTSThe expression rate of CD80 and CD83 in the emodin group were 13.4% +/- 6.6% and 9.3% +/- 2.2% respectively; which were significantly lower than those in the control group (39.3% +/- 8.6% and 30.7% +/- 5.6%), respectively (P<0.05). IL-12 secretion of DCs was lower (1700.44 +/- 1000.21 microg/L vs 4500.60 +/- 1200.6 microg/L) but IL-10 secretion was higher (350.6 +/- 150.2 microg/L vs 230.7 +/- 90.1 microg/L) in the emodin group than in the control group (P<0.05). Mixed lymphocyte culture (MLR) examination showed that emodin could significantly inhibit the stimulation of DCs on self-T-lymphocyte proliferation.

CONCLUSIONEmodin could evidently suppress the maturation and immune stimulating function of DCs during their in vitro conversion process.

Cell Differentiation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; Emodin ; pharmacology ; Humans

Animals ; Apoptosis/drug effects* ; Autophagy/drug effects* ; Carnitine/pharmacology* ; Cells, Cultured ; Endothelial Cells ; Lipopolysaccharides ; Mice

No abstract available.
Animal ; Apoptosis/*drug effects ; Human ; Tumor Cells, Cultured

OBJECTIVESTo investigate the effects of norepinephrine (NE) on the proliferation and apoptosis of hepatic stellate cells (HSCs).

METHODSCultured HSCs were used in 6 groups: (1) a control group; (2) a NE group; (3) a phentolamine plus propranolol group; (4) a phentolamine (an alpha-AR antagonist) group; (5) a CGP20712A (a beta1-AR antagonist) group; and (6) a ICI118551(a beta2-AR antagonist) group. After NE and the antagonists of adrenoceptor subtypes were administered to the cultured HSCs, MTT assay was used to evaluate the cell proliferation at 24 h, 48 h, and 72 h. Terminal deoxyribonucleotidyltransferase-mediated dUTP nick end labelling (TUNEL) assay and flow cytometry were used to detect cell apoptosis. An inverted microscope was used to observe the morphological changes of HSCs.

RESULTS(1) MTT assay indicated that NE significantly induced HSCs proliferation in a time-dependent manner, which were reduced by antagonist of alpha-AR, beta1-AR and beta2-AR. (2) At 24 h after HSCs exposure to NE, apoptosis rates decreased significantly compared with that of the control group (6.60%+/-3.05% vs 12.60%+/-4.76%). In the antagonists of adrenoceptor subtypes groups, especially of a and beta2 adrenoceptor subtypes, the apoptosis was less. (3) Apoptosis rate of the NE group was significantly lower than that of the control group (2.29%+/-0.22% vs 3.06%+/-0.57%). In the antagonists of alpha and b2 adrenoceptor groups the apoptosis was less. (4) No obvious morphological changes of HSCs were found after administration of NE.

CONCLUSIONSSympathetic neurotransmitter NE can induce proliferation and inhibit apoptosis of the cultured HSCs.

Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Hepatic Stellate Cells ; cytology ; drug effects ; Norepinephrine ; pharmacology ; Rats

OBJECTIVETo analyze the influence of retinoic acid (RA) on the undifferentiated state and EB formation abilities of human embryonic stem cells.

METHODSThe biological characteristics of H9 ESCs after RA treatment were characterized by real-time PCR, MTS proliferation assay and immunofluorescence staining. The expression of three germ layers markers, osteogenic differentiation markers and adipogenic differentiation markers in H9-differentiated embryoid bodies (EBs) with RA treatment were quantified by real time PCR.

RESULTSThe proliferation of H9 ESCs in the early logarithmic growth phase was accelerated by RA treatment. In addition, RA induced differentiation of H9 ESC coupled with morphology changes, decreased expression of undifferentiated markers Oct4, Nanog, Sox2 and OCT4 mRNA binding protein Lin28 at mRNA level, and reduced expression of Oct4 at protein level. RA induced formation of cavities in EBs. Real time PCR results showed that the expressions of ectodermal markers: NeuroD1, Noggin; mesodermal markers: Brachyury, Twist and endodermal markers: AFP, GATA-4 were significantly increased (P < 0.05), especially for AFP (P < 0.01), by RA treatment in a dose-dependent manner. In addition, the expression of adipogenic differentiation marker C/EBPalpha was increased while the osteogenic differentiation marker OPN was decreased in EBs after RA treatment for 5 days.

CONCLUSIONSHigh concentrations of RA induced the loss of stemness in H9 ESCs and excessive differentiation in EBs, and damaged the balance between osteogenic and adipogenic differentiation during early EB differentiation, which may be relevant to the congenital malformations.

Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; drug effects ; Humans ; Tretinoin ; pharmacology

OBJECTIVETo investigate the effects of ephedrine on human nasal cilia movement.

METHODSCiliary beat frequency (CBF) of cultured human nasal epithelial cells was measured by high-speed digital microscopy in HBSS and ephedrine solution of different concentrations in 10 minutes.

RESULTSCBF of cultured nasal epithelial cells exposed to HBSS showed no significant changes in 10 minutes. However, in 2.5 g/L , 5 g/L, 10 g/L and 20 g/L ephedrine solution, CBF increased significantly in 1-2 minutes and reached the apex, then it decreased gradually, at the 10th minute. CBF of the samples exposed to 2.5 g/L and 5 g/L ephedrine solution were slower than those in HBSS, but no significant changes were found. However, in 10 g/L and 20 g/L ephedrine solution, CBF decreased significantly when compared with samples in sHBSS. With the concentrations from 2.5 g/L to 20 g/L ephedrine, the increment was independent on the concentration, the inhibitory effect was dependent on the concentration.

CONCLUSIONSIn initial time, 2. 5 g/L-20 g/L ephedrine stimulated CBF, then 10 g/L-20 g/L ephedrine inhibited CBF. The stimulation of 2.5 g/L and 5 g/L ephedrine on CBF was longer than that of 10 g/L and 20 g/L ephedrine. 5 g/L ephedrine had maximum stimulatory effect without obvious inhibitory effect on cultured human nasal CBF.

Cells, Cultured ; Cilia ; drug effects ; physiology ; Ephedrine ; pharmacology ; Epithelial Cells ; drug effects ; physiology ; Humans ; Nasal Mucosa ; cytology ; drug effects ; physiology

Endothelial progenitor cells (EPCs) play an important role in diabetic vascular complications. A large number of studies have revealed that some clinical antihyperglycemics can improve the complications of diabetes by regulating the function of EPCs. Metformin can improve EPCs function in diabetic patients by regulating oxidative stress level or downstream signaling pathway of adenosine monophosphate activated protein kinase; Pioglitazone can delay the aging of EPCs by regulating telomerase activity; acarbose, sitagliptin and insulin can promote the proliferation, migration and adhesion of EPCs. In addition to lowering blood glucose, the effects of antihyperglycemics on EPCs may also be one of the mechanisms to improve the complications of diabetes. This article reviews the research progress on the regulation of EPC proliferation and function by antihyperglycemics.
Cell Movement/drug effects* ; Cells, Cultured ; Endothelial Progenitor Cells/drug effects* ; Humans ; Hypoglycemic Agents/pharmacology* ; Signal Transduction/drug effects*

OBJECTIVETo evaluate possible inactivating effect of recombined decoction in on mumps virus.

METHODSBy adopting tissue cell culturing technology, a group of viruses including the mumps virus, herpes simplex virus (type I, II), rubella virus, cytomegalovirus (CMV), herpes zoster virus, influenza virus, parainfluenza virus, adeno viruses, respiratory syneytial virus (RSV) were cultured. The cells infected with the viruses were treated with the decoction.

RESULTSThe decoction showed remarkable inhibitory and killing effects on the mumps virus while had no obvious inhibitory and killing effects on host's cells, herpes simplex virus (type I, II), rubella virus, cytomegalovirus (CMV), herpes zoster virus, influenza virus, parainfluenza virus, adeno viruses, respiratory syneytial virus (RSV).

CONCLUSIONSThe decoction had obvious inhibitory and killing effects on mumps virus during single layer cells culture.

Cells, Cultured ; Cytomegalovirus ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Mumps virus ; drug effects ; Respiratory Syncytial Viruses ; drug effects ; Respirovirus ; drug effects ; Rubella virus ; drug effects ; Simplexvirus ; drug effects

Effects of lysophosphatidic acid (LPA), an extracellular phospholipid signal, on the proliferation of rat embryonic neural stem cells (NSCs) and their differentiation into microtubule-associated protein 2 (MAP2)-positive and choline acetyltransferase (ChAT)-positive, i.e. cholinergic-committed neurons, were observed in vitro by [(3)H]-thymidine incorporation, immunocytochemistry, Western blot and other techniques. The results showed that: (1) Lower concentrations of LPA (0.01~1.0 mumol/L) dose-dependently enhanced the uptake of [(3)H]-thymidine by NSCs cultured in specific serum-free medium, indicating a significant promotive action of LPA on the proliferation of NSCs. (2) After fetal bovine serum which induces and commences the differentiation of NSCs, was used in the medium, the lower concentrations of LPA increased the percentages of both MAP2- and ChAT-immunoreactive neurons, with a peak at 0.1 mumol/L LPA in two cases. (3) The promotive effects of LPA on the differentiation of MAP2- and ChAT-positive neurons were also supported by the up-regulation of the expressions of both MAP2 and ChAT proteins detected by Western blot. (4) At the early phase of differentiation of NSCs, the cell migration and neurite extension were enhanced significantly by lower dosages of LPA under phase-contrast microscope. These results suggest that LPA within certain lower range of concentrations promotes the proliferation of NSCs and their differentiation into unspecific MAP2-positive and specific cholinergic-committed neurons, and also strengthens the migration and neurite extension of the newly-generated neuronal (and also glial as reported elsewhere) progenitors.
Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cholinergic Neurons ; cytology ; Embryonic Stem Cells ; drug effects ; Lysophospholipids ; pharmacology ; Neural Stem Cells ; drug effects ; Rats

BACKGROUNDBisphosphonates (BPs) have been reported to reduce local recurrence in giant cell tumor (GCT) of bone because of their osteoclast-suppressing effect; however, the optimal mode of delivery and the dose and duration of treatment of BPs remain to be established. To address these issues, it is first necessary to clarify the manner of action of BPs on osteoclasts. We herein evaluated the osteoclast-suppressing effect of sodium ibandronate in vitro.

METHODSMouse osteoclasts (OCLs) were generated in vitro using mouse bone marrow mononuclear cells. First, various concentrations of sodium ibandronate and equal amounts of phosphate-buffered saline were added to cell culture media. The number of multinucleated cells (over three nuclei) was recorded in each group, OCL formation was compared, and the most effective concentration of sodium ibandronate was determined. Then, high concentrations of sodium ibandronate were added to the experimental cell culture media; no ibandronate was given in the control group. Comparisons were made between the two groups in terms of OCL adhesion, migration, and bone resorption.

RESULTSOCL formation was suppressed by sodium ibandronate in vitro; the most pronounced effect was observed at the concentration of 10(-5) mol/L. OCL migration and bone resorption were significantly suppressed at this concentration, though there was no effect on OCL adhesion.

CONCLUSIONSSodium ibandronate was effective in suppressing OCLs and decreasing resorption in GCT. The strong anti-OCL effectiveness at a high concentration in vitro indicates a topical mode of application.

Animals ; Bone Resorption ; Cell Movement ; drug effects ; Cells, Cultured ; Diphosphonates ; pharmacology ; Mice ; Osteoclasts ; cytology ; drug effects