OBJECTIVETo evaluate in vitro effect of abnormal savda munziq (ASMq) on the proliferation and apoptosis of human hypertrophic scar fibroblasts (HSFs).

METHODSHSFs were divided into six groups to receive different treatments as group A (blank control group), group B-E (ASMq in different concentration), and group F(5-Fu). Each group contains six specimens. The HSFs were cultured in vitro. After culture for 48 hours, the CCK8 test and flow cytometry methods were used to detect the proliferation, cell cycle and apoptosis.

RESULTSThe proliferation of HSFs in the B, C, D and E groups was inhibited at G2/M period, while it was inhibited at G0/S period in group F (P < 0.05). The inhibition effect of ASMq (0.1-1.0 mg/ml) on the fibroblasts enhanced in a concentration-dependent manner. Flow cytometry analysis with annexin V-FITC and PI staining confirmed the apoptotic. When HSFs were exposed to ASMq at 1.0 mg/ml (group E) for 48 h, the percentage of apoptotic cells increased to (43.7 +/- 2.58)%, which was significantly higher than that of blank control group (2.2 +/- 0.59)%. The induced apoptosis effect was also increased in a concentration-dependent manner.

CONCLUSIONASMq has a inhibitory effect on the proliferation and an enhancement effect on the apoptosis of fibroblast. ASMq could be used as an effective drug for treatment of hypertrophic scar.

Apoptosis ; Cell Cycle ; drug effects ; physiology ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; pathology ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; In Vitro Techniques ; Medicine, East Asian Traditional

OBJECTIVETo compare the mechanisms of G(2)/M cycle arrest induced by topo IIalpha and IIbeta inhibitors in H460 cells.

METHODSThe inhibitory effects of XK469, adriamycin and etoposide on H460 cell growth were analyzed by MTT assay. The changes in cell cycle and expressions of cdc2, phos-cdc2 and 14-3-3sigma proteins induced by these 3 topo II inhibitors were detected by flow cytometry and Western blotting, respectively.

RESULTSBoth of the two types of topo II inhibitor resulted in dose-dependent G(2)/M phase arrest and growth inhibition of H460 cells, but XK469 failed to induce 14-3-3sigma protein expression as adriamycin and etoposide did.

CONCLUSIONTopo IIalpha and topo IIbeta inhibitors induce growth inhibition of H460 cells possibly through two different mechanisms, namely the 14-3-3sigma-dependent pathway and the 14-3-3sigma-independent pathway, but further functional inhibition test of 14-3-3sigma is needed to confirm this hypothesis.

Antigens, Neoplasm ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Cycle ; drug effects ; physiology ; Cell Division ; drug effects ; Cell Line, Tumor ; DNA Topoisomerases, Type II ; DNA-Binding Proteins ; antagonists & inhibitors ; G2 Phase ; Humans ; Lung Neoplasms ; pathology ; Quinoxalines ; pharmacology ; Topoisomerase II Inhibitors

OBJECTIVETo evaluate the effect of enamel matrix protein (EMP) on the attachment and proliferation of periodontal ligament cells (PDLC) on diseased cementum surfaces in vitro.

METHODSCementum chips were obtained from diseased roots exposed to periodontal pocket. Thirteen diseased root cementum chips were conditioned with EMP. Meanwhile, 13 diseased and 13 healthy cementum chips were treated with physiological saline as control. The growth and morphology of PDLC on the root surface were observed after 24 hours incubation by scanning electron microscope (SEM). PDLC attachment and proliferation were quantified using MTT assay at 16 or 72 hours.

RESULTSThe cells on EMP treated roots under SEM were growing robust like the cells on healthy roots. By contrast, the diseased cementum surface without conditioned with EMP was only partly covered with spindle-shaped cells, with filopodia appearing short and thin. MTT assay indicated that the number of adhered and proliferated cells on diseased cementum chips treated with EMP was significantly greater than that on diseased chips treated with saline (adhesion: 0.45 +/- 0.03 vs. 0.37 +/- 0.05, P < 0.05; proliferation: 0.71 +/- 0.02 vs. 0.55 +/- 0.08, P < 0.01), but less than that on healthy chips (adhesion: 0.45 +/- 0.03 vs. 0.67 +/- 0.08, P < 0.05; proliferation: 0.71 +/- 0.02 vs. 1.05 +/- 0.09, P < 0.05).

CONCLUSIONSIt was suggested that EMP could promote the growth of PDLC on the diseased root cementum surface.

Cell Adhesion ; drug effects ; Cell Division ; drug effects ; Cells, Cultured ; Dental Cementum ; physiology ; Dental Enamel Proteins ; pharmacology ; Humans ; Microscopy, Electron, Scanning ; Periodontal Ligament ; cytology ; Periodontitis ; pathology

Neuronal PC12 cells induced by nerve growth factor (NGF) have been considered to be postmitotic and lack the ability to divide. However, in this study, we not only detected DNA synthesis but also observed cell division in some morphologically differentiated neuronal PC12 cells bearing long neurites. More interestingly, in addition to the division of perikaryon, the neurites located on the division site of the cell membrane also divided into two parts and were allocated to the two daughter cells. These results demonstrate that the morphologically differentiated neuronal PC12 cells still retain the ability to divide. This is the first report that neuronal PC12 cells as well as their neurites can divide.
Animals ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; DNA Replication ; drug effects ; physiology ; Nerve Growth Factor ; pharmacology ; Neurites ; drug effects ; Neurons ; cytology ; PC12 Cells ; Rats

OBJECTIVELiver development needs a number of growth factors and components of the extracellular matrix. The study is to explore how growth factors and extracellular matrix regulate proliferation and differentiation of fetal liver progenitor cell.

METHODSWe demonstrate isolation of hepatic progenitor/stem cells from ED 14.5 SD rat liver, which contains a large number of hepatoblasts. Proliferation assay-3H thymidine incorporation was used to detect the effect of growth factors on proliferation of hepatic progenitor cell. Growth factor and extracellular matrix were added and stem cell clone formation was counted. Mark of bile duct and hepatocyte were detected with double-marker immunocytochemistry.

RESULTSProgenitor liver cells displayed clonogenic capacity, expressed markers of hepatocytes and bile duct cells and G-6-P. HGF, EGF can accelerate DNA synthesis and stem cell clone formation of hepatic progenitor cell. Extracellular matrix collagen I, collagen IV or laminin were essential for formation of stem cell clone. Single cell culture needed HGF, EGF, extracellular matrix and supernatant of mix cell (which contained fetal parenchymal cells, mesenchymal cells and hematopoietic cells) culture.

CONCLUSIONGrowth factors especially HGF and EGF play crucial role in proliferation and differentiation of liver progenitor cell. Some factors secreted from mesenchymal cell and hematopoietic cells may be involved.

Animals ; Cell Differentiation ; drug effects ; Cell Division ; physiology ; Cells, Cultured ; Epidermal Growth Factor ; pharmacology ; Extracellular Matrix ; physiology ; Female ; Fetus ; Hepatocyte Growth Factor ; pharmacology ; Liver ; cytology ; drug effects ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; drug effects

Antineoplastic Agents ; pharmacology ; Bile ; physiology ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; Celecoxib ; Cell Division ; drug effects ; Cell Line, Tumor ; Cholangiocarcinoma ; metabolism ; pathology ; Cyclooxygenase 2 ; Dinoprostone ; metabolism ; Humans ; Isoenzymes ; biosynthesis ; genetics ; Membrane Proteins ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; Pyrazoles ; RNA, Messenger ; genetics ; Sphincterotomy, Transduodenal ; adverse effects ; Sulfonamides ; pharmacology ; Up-Regulation

OBJECTIVETo investigate the neointima formation and the expression of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-alpha (TNF-alpha) in cuff-induced vascular injury in mouse model, and to examine the effect of angiotensin II type 1 receptor (AT1) blocker, olmesartan, on MCP-1 and TNF-alpha expression and consequently vascular remodeling.

METHODSVascular injury was induced by polyethylene cuff-placement around the mouse femoral artery. Some mice were treated with AT1 receptor blocker, olmesartan, at the dose of 3 mg.kg(-1).day(-1) with an osmotic minipump. Neointima formation and the proliferation of vascular smooth muscle cells (VSMCs) were measured by morphometric analysis and bromodeoxyuridine (BrdU) incorporation. MCP-1 and TNF-alpha expression was detected by Western blot and immunohistochemical staining.

RESULTSWe observed neointima formation 14 days after cuff placement as well as VSMCs proliferation in the media and neointima. Cuff placement also induced MCP-1 and TNF-alpha expression in the media and neointima that the VSMCs specifically existed. Treatment of mice with olmesartan at a dose of 3 mg.kg(-1).day(-1), which did not influence systolic blood pressure, significantly decreased neointima formation and the proliferation of VSMCs. Olmesartan also inhibited MCP-1 and TNF-alpha expression in the injured arteries.

CONCLUSIONSOur results demonstrate that blockade of AT1 receptor inhibits MCP-1 and TNF-alpha expression and thereby improves vascular remodeling.

Analysis of Variance ; Animals ; Blotting, Western ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Chemokine CCL2 ; analysis ; Disease Models, Animal ; Imidazoles ; pharmacology ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred C57BL ; Monocytes ; cytology ; drug effects ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Neovascularization, Physiologic ; drug effects ; physiology ; Olmesartan Medoxomil ; Probability ; Sensitivity and Specificity ; Tetrazoles ; pharmacology ; Tumor Necrosis Factor-alpha ; analysis ; drug effects ; Tunica Intima ; drug effects ; pathology ; Vascular Diseases ; physiopathology

OBJECTIVETo investigate the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid (HA) on the proliferation of rabbit chondrocytes in vitro.

METHODSChondrocytes from the knee joints of New Zealand white rabbits were cultured. bFGF or HA or both were added into the culture medium respectively, and the proliferation of the chondrocytes was measured with MTT 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyl-tetra-zolium bromide. (MTT, Sigma, M2128).

RESULTSBasic fibroblast growth factor (10 ng/ml) with low concentration of fetal bovine serum in the culture medium promoted the proliferation of chondrocytes significantly, and this effect reached its maximum when concentration of bFGF reached 50 ng/ml. HA itself had no effect on the proliferation of chondrocytes. However, when bFGF was used in combination with HA, especially when the concentration of bFGF was 50-500 ng/ml and that of HA was 10-50 ng/ml, the effect on the proliferation of chondrocytes was much more than when bFGF or HA was used alone.

CONCLUSIONSbFGF can promote the proliferation of chondrocytes. HA, which has no effect on the proliferation of the cells, can maintain a normal growth of chondrocytes. When bFGF is used in combination with HA, more proliferation is obtained.

Analysis of Variance ; Animals ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Chondrocytes ; drug effects ; physiology ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Hyaluronic Acid ; pharmacology ; In Vitro Techniques ; Knee Joint ; cytology ; Male ; Probability ; Rabbits ; Sensitivity and Specificity

OBJECTIVETo investigate the regulatory effect of IFN-gamma on recognition of target cells by human natural killer (NK) cells.

METHODSThe cytotoxic activity of human NK cell lines (NK92, NKL) was detected by MTT method. Expression of NK cell receptors (NKG2D, NKG2A/B, KIR2DL1 and KIR2DS1) and MICA on target cells (the ligand of NKG2D) was measured by RT-PCR.

RESULTSBoth NK92 and NKL cells exerted higher cytotoxicity to tumor cells with MICA expression, while tumors without MICA expression could resist NK cell lysis. IFN-gamma (> 1000 U/ml) inhibited NK lysis of tumor cells with MICA expression through down-regulating the expression of NKG2D, but up-regulating the expression of NKG2A/B and KIR2DL1.

CONCLUSIONIFN-gamma has a negative effect on activation and cytotoxicity of human NK cells by altering the balance between the expression of activating and inhibitory receptors on NK cells in favor of inhibition. This may serve to limit NK cell over-activation in vivo.

Cell Division ; drug effects ; Cytotoxicity, Immunologic ; drug effects ; Histocompatibility Antigens Class I ; analysis ; physiology ; Humans ; Interferon-gamma ; pharmacology ; Killer Cells, Natural ; immunology ; NK Cell Lectin-Like Receptor Subfamily C ; NK Cell Lectin-Like Receptor Subfamily K ; Receptors, Immunologic ; metabolism ; Receptors, KIR2DL1 ; Receptors, Natural Killer Cell ; Recombinant Proteins ; Tumor Cells, Cultured

OBJECTIVETo determine the modulating effect of CD40 ligand (CD40L, CD154) on CD40-transfected human lung carcinomas and to assess the potential of CD40 as a therapeutic target.

METHODSTumor cells of a CD40-negative lung cancer cell lines (GLC-82) were transfected with a vector expressing CD40 cDNA. The transfected cell line, GLC-82/CD40, was shown to express high levels of CD40. GLC-82/CD40 cells after being exposed to 0.1 micro g/ml CD40L were examined for their surface expression, cell cycle, apoptosis and cell growth by flow cytometry and MTT assay.

RESULTSThe expression of MHC-I, ICAM-1 and Fas in GLC-82/CD40 cells was significantly increased, whereas that of EGFR was decreased. Cell proliferation was significantly inhibited by CD40L with an inhibition rate of 30% on day 5, but no change in cell cycle. All of the changes disappeared after 48 h incubation with CD40L. More significant changes were observed in Calu-3 cell lines which expressed high levels of CD40, but the CD40-negative GLC-82 cells were unresponsive to CD40L. None of the 3 cell lines showed significant changes in apoptosis upon CD40L treatment.

CONCLUSIONCD40, if over-expressed in tumor cells, could be considered as a potential therapeutic target.

Apoptosis ; drug effects ; Blotting, Western ; CD40 Antigens ; physiology ; CD40 Ligand ; pharmacology ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Transfection