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

OBJECTIVETo investigate the effect of the basic fibroblast growth factor (b-FGF) to regenerate an autologous tissue-engineered cartilage in vitro.

METHODSThe Cells were harvested from the elastic auricular cartilage of swine,and were plated at the concentration of 1 x 10(4) cells/cm2 , studied in vitro at two different media enviroments: Group I contained Ham's F-12 with supplements and b-FGF, Group II contained Ham's F-12 only with supplements. The passage 2 cells (after 12.75 +/- 1.26 days) were harvested and mixed with 30% pluronic F-127/Ham's F-12 at the concentration of 50 x 10(6) cells/ml. It was injected subcutaneously at 0.5 ml per implant. The implants were harvested 8 weeks after the vivo culture and examined with the histological stains.

RESULTSThe chondrocytes displayed morphologically similar to the fibroblasts in the media containing basic-FGF. The number of cell doublings (after 12.75 +/- 1.26 days) in vitro culture was as the following: Group I, 70; Group II, 5.4. Eight 8 weeks after the vivo autologous implantation, the average weight (g) and volume (cm3) in each group was as the following: Group I, 0.371 g/0.370 cm3 Group II, 0.179 g/0.173 cm3 (P < 0.01). With the b-FGF in vitro culture, the cells were expanded by 70 times after 2 weeks. Histologically, all of the engineered cartilage in the two groups were similar to the native elastic cartilage.

CONCLUSIONThese results indicate that the basic-FGF could be used positively to enhance the quality and quantity of the seeding cells for the generation of the well-engineered cartilage.

Animals ; Cartilage ; cytology ; drug effects ; physiology ; Cell Division ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; Female ; Fibroblast Growth Factors ; pharmacology ; physiology ; Male ; Regeneration ; drug effects ; Swine ; Tissue Engineering ; methods ; Transplantation, Autologous

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

OBJECTIVETo investigate the expression of apoptotic protein inhibitors, survivin and XIAP, in patients with myelodysplastic syndromes (MDS) and in the cell line MUTZ-1, as well as to explore the possible mechanisms of homoharringtonine (HHT) in the treatment of MDS.

METHODSBone marrow samples from 47 patients with de novo MDS at diagnosis were examined and bone marrow samples from 15 normal donors were used as control. A MDS-RAEB cell line MUTZ-1 was used as in vitro model. Detection of apoptotic cells and cell cycle analysis were performed with flow cytometry (FACS). The expression of apoptotic protein inhibitor survivin and XIAP in the MDS cells were detected by RT-PCR technique. MUTZ-1 were treated with antisense oligodeoxynucleotide (AS-ODNs) of survivin and or HHT, the effects were evaluated by cell viability and cell apoptosis.

RESULTSSurvivin mRNA positive rate in MDS were significantly higher than that in normal controls (38.3% and 0, respectively, P < 0.01), and the positive rate in high risk group (RAEB, RAEBT and CMML) was significantly higher than that in RA/RAS group (53.6% and 16.7%, respectively, P < 0.05). XIAP was expressed in all untreated MDS and healthy controls. XIAP mRNA expression in high risk group was significantly higher than that in RA/RAS subtypes and healthy controls (1.55 +/- 0.34, 0.74 +/- 0.24, and 1.01 +/- 0.28, respectively, P < 0.01). However, XIAP mRNA expression was significantly lower in RA/RAS subtypes than in healthy control (0.74 +/- 0.24 and 1.01 +/- 0.28, P < 0.054). Apoptosis peak detected by FACS analysis and positive Annexin V FITC staining on cell membrane indicated that HHT could induce MUTZ-1 cell undergoing apoptosis in dose- and time-dependent manners. Treatment of MUTZ-1 cells with HHT revealed that HHT could significantly down-regulate survivinexpression but had no significant effect on XIAP expression in the cells. AS-ODNs of survivin could inhibit MUTZ-1 cells growth, induce them to apoptosis and sensitize them to HHT.

CONCLUSIONThe expression levels of survivin; Institute of Hematology, Oncology and Tumor Immunology, Robert Roessle Clinic, Humboldt University, Berlin, Germany (Wolf Dieter Ludwig, Christian Wuchter) and XIAP vary in different subtypes of MDS patients, suggesting that the proteins may play an important role in the pathogenesis of the disease. Down-regulation of survivin in MUTZ-1 cells may be one of the mechanisms that HHT induces apoptosis of MDS cells.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Harringtonines ; therapeutic use ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; physiology ; Myelodysplastic Syndromes ; drug therapy ; pathology ; Neoplasm Proteins ; Oligonucleotides, Antisense ; pharmacology ; Proteins ; genetics ; physiology ; RNA, Messenger ; analysis ; X-Linked Inhibitor of Apoptosis Protein

OBJECTIVETo study the effects of c9,t11-conjugated linoleic acid on the killing ability of macrophage to B16-MB cells in C57 mice and explore its possible mechanism.

METHODSThe five levels of CLA was designed as 0, 25, 50, 75, 100 micro mol/L. After macrophage was treated with CLA for 24 h, the killing ability of macrophage on B16-MB cells was evaluated by MTT, The expression of C57 mice macrophage cytokine IL-6, TNF-alpha and iNOS mRNA was detected by RT-PCR. The expression of Erk protein was examined by Western Blot assay.

RESULTSThe inhibitory effect of macrophage on tumor cell depend on the treatment of the increased c9,t11-CLA level, at the same time, the expression of IL-6, TNF-alpha and iNOS mRNA increased, the expression of Erk decreased with the elevating dose of CLA.

CONCLUSIONSc9,t11-CLA could increase the killing ability of macrophage in mice to B16-MB cells, and it was associated with induction of IL-6, TNF-alpha and iNOS mRNA expression. We speculate that antitumor ability of CLA may be associated with taking part in body immune regulation action, and the effects of CLA on the killing ability of murine macrophage to B16-MB cells was not associated with the MAPKErk pathway.

Animals ; Blotting, Western ; Cell Division ; Cell Line, Tumor ; Coculture Techniques ; Dose-Response Relationship, Drug ; Interleukin-6 ; genetics ; Linoleic Acids, Conjugated ; pharmacology ; Macrophages ; drug effects ; physiology ; Melanoma, Experimental ; pathology ; Mice ; Mice, Inbred C57BL ; Mitogen-Activated Protein Kinases ; metabolism ; Nitric Oxide Synthase ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo observe the effect of Erzhi Tiangui recipe (ETR) on quality of oocyte in the process of external fertilization and embryo-transplantation.

METHODSEighty mice were randomly divided into 4 groups, Group A treated with ETR plus human menopausal gonadotropin (HMG), Group B with ETR, Group C with HMG and Group D with normal saline. Ovulation test and cleavage test were conducted to observe the effect of treatment on quality of oocytes.

RESULTSThe difference on ovulation number between Group A and C was insignificant, but the difference in comparison between the two groups was significant in aspects of oocyte morphological scoring, fertilization rate and cleavage rate (P<0.05).

CONCLUSIONETR could play its effect synergistically with Western medicine, and raise the quality of oocytes.

Animals ; Cell Division ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Embryo Transfer ; Female ; Fertilization ; drug effects ; Fertilization in Vitro ; drug effects ; Menotropins ; pharmacology ; Mice ; Oocytes ; drug effects ; physiology ; Ovulation Induction ; Random Allocation

In order to explore the underlying mechanism of high effects and low toxicity of trichostatin A (TSA), the effect of TSA on growth inhibition, histone acetylation level and apoptosis in HL-60 cells and normal human peripheral blood mononuclear cells (NPBMNC) were examined using MTT method, immunocytochemistry technology, and Annexin-V-FITC/PI double staining flow cytometry. The results showed that TSA inhibited growth of HL-60 in time- and dose-dependent manners, and the IC(50) of 36 hours was 100 ng/ml. The apoptosis induction effect of TSA in HL-60 cells was also time- and dose-dependent. Besides, the dose of TSA showing significant apoptotic cytotoxicity in HL-60 cells did not demonstrate apparent apoptosis induction in NPBMNC within definite dose and time range. The histone acetylation level in HL-60 cells and NPBMNC both showed remarkable increase (P < 0.05) after incubated with 100 ng/ml TSA for 4 hours without statistical difference between them is detected (P > 0.05). It is concluded that TSA shows effects of definite and significant growth inhibition and apoptosis induction on HL-60 cells in time- and dose-dependent manners. TSA is able to selectively induce apoptosis in HL-60 cells with low toxicity in NPBMNC at the same time. The mechanism of this selectivity can not be ascribed to the differential regulation of histone acetylation level between HL-60 cells and NPBMNC.
Acetylation ; Apoptosis ; drug effects ; Cell Division ; drug effects ; DNA-Binding Proteins ; HL-60 Cells ; drug effects ; Histone Deacetylases ; physiology ; Histones ; metabolism ; Humans ; Hydroxamic Acids ; pharmacology ; RNA, Messenger ; analysis ; Telomerase ; genetics