OBJECTIVETo study radiation-enhancing effects on human gastric cancer MKN28 cell line and underlying mechanisms of β-elemene.

METHODSInhibition of MKN28 cell proliferation at different concentrations of β-elemene was assessed using the methyl thiazolyl blue colorimetric method (MTT method), with calculation of IC50 value and choice of 20% of the IC50 as the experimental drug concentration. Irradiation group and β-elemene+irradiation group were established, and the cell survival fraction (SF) was calculated from flat panel colony forming analysis, and fitted by the 'multitarget click mathematical model'. Draw the survival curve and get the radiobiological parameters D0, Dq, SF2, N and SER. Flow cytometry (FCM) was used to detect changes in the cell cycle and cell apoptosis rates was detected by Annexin-V/PI assay.

RESULTSβ-elemene exerted inhibitory effects on proliferation of gastric cancer MKN28 cells, with an IC50 of 45.6 mg/L and we chose 8 mg/L as the experimental concentration. The cell survival fraction of MKN28 cells with irradiation decreased significantly after treated with β-elemene; D0, Dq decreased, SER = 1.3. After combined treatment of β-elemene+irradiation, the results of FCM showed that cells could be arrested in the G2/M phase and the cell apoptosis increased significantly.

CONCLUSIONSβ-elemene can enhance the radiosensitivity of gastric cancer MKN28 cell line. Mechanistically, β-elemene mainly influences the cell cycle distribution of MKN28 cells by inducing G2/M phase arrest, inhibits the repair of sublethal damage and induces cell apoptosis to enhance the killing effects of radioactive rays.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Humans ; Radiation Tolerance ; drug effects ; Sesquiterpenes ; pharmacology ; Stomach Neoplasms ; pathology

The human fibroblast MRC-5 cells incubated with PHB granules (TM) added at a final concentration of 4 mg/ml showed a time-course pattern of survival. The percentages of dead cells obtained were at the rate of 3.8% after 7 days, respectively. When the MRC-5 cells grown in different material, using the test concentration of 4 mg/ml PCM, they were found to show a similar time-course increasing pattern of death as that obtained with PHB. However, the death was noted in the cells incubated for 7 days, the death rates obtained was 40.54% respectively.
Cell Line ; Cell Survival/*drug effects ; Fibroblasts/drug effects ; Hydroxybutyrates/*toxicity ; *Materials Testing ; Polymers/*toxicity

OBJECTIVETo study the maturation effect of CpG2006 and phosphodiester oligonucleotides on leukemia-derived dendritic cells.

METHODSLeukemia cells K562/A02 were induced into dendritic cells by rhGM-CSF and rhIL-4. After 7 days induction, the cell-morphology was observed, the immunophenotype of cells was detected by flow cytometry and the cell function was evaluated by allogeneic mixed lymphocyte reactions, CTL responses and secretion of IL-12 and IL-6. Then a CpG oligonucleotide CpG2006, two synthetic bacterial phosphodiester oligonucleotides A-ODN and T-ODN were added to these leukemia-derived DCs. Three days later, the DCs were re-detected by the above-mentioned methods.

RESULTSAfter induced by CpG2006, A-ODN or T-ODN, the leukemia-derived DCs with typical dendritic morphology were increased. The expressions of CD83, HLA-DR and CD86 were (65.5 +/- 8.4)%, (32.0 +/- 4.3)% and (18.6 +/- 3.2)% respectively in day 7 leukemia-derived DCs, raised to (88.9 +/- 3.6)%, (53.9 +/- 3.2)% and (39.9 +/- 7.3)% respectively after exposing CpG2006 for 3 days; increased to (97.0 +/- 5.3)%, (63.9 +/- 7.3)% and (40.2 +/- 7.4)% respectively after treated by A-ODN; and further increased to (93.26 +/- 4.65)%, (58.3 +/- 5.6)% and (36.2 +/- 6.8)% respectively after treated by T-ODN. These results was markedly different than unaffected cells did. These DCs induced by the above-mentioned three oligonucleotides could upregulate significantly the capacity for stimulating allogeneic T cells. They could also induce CTL to generate specific cytotoxic activity against K562/A02 cells. The secretion of IL-6 and IL-12 was increased remarkably.

CONCLUSIONCpG2006, as well as two phosphodiester oligonucleotides can induce leukemia-derived DCs maturation.

Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Dendritic Cells ; cytology ; drug effects ; Humans ; K562 Cells ; Oligodeoxyribonucleotides ; pharmacology ; Oligonucleotides ; pharmacology

To improve the function of endothelial progenitor cells (EPCs) is one of the goals in Chinese traditional therapy to treat various cardio-celebrovascular diseases. In the past decades, scholars in the field of traditional Chinese medicine (TCM) have found fifteen active compounds to regulate the function of EPC. These metabolites are extracted from thirteen, plant-based Chinese medicine, with majority of them as potent reductive or oxidative hydrophilic molecules containing phenyl groups. These active compounds either enhance the mobilization of EPC, or inhibit their apoptosis through different signaling pathways. In this review, the molecular structure, biophysical properties, and the plant sources of these active ingredients and their regulatory effects on the function of EPC are summarized, aiming to reveal the modern basis of Chinese medicine for promoting blood circulation and removing blood stasis at the progenitor cell level.
Animals ; Apoptosis ; drug effects ; Cell Movement ; drug effects ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Progenitor Cells ; cytology ; drug effects ; metabolism ; Humans ; Signal Transduction ; drug effects

OBJECTIVESTo analyze the changes of movement function and viability in human spermatozoa induced by reactive oxygen species(ROS), and to prove whether ROS were one of the causes of the movement dysfunction of spermatozoa.

METHODSSpermatozoa with normal physiological functions selected from semen samples by Percoll gradient centrifugation technique were regarded as normal sperm models. ROS were generated by hypoxanthine-xanthine oxidase system and then incubated with normal sperm models under aerobic environment. After model spermatozoa were incubated with ROS, movement parameters of spermatozoa were analyzed by computer-assisted semen analysis (CASA) system.

RESULTSCompared with the control group, after model spermatozoa were incubated with ROS for 30 minutes, motility, curvilinear velocity(VCL), straight line velocity (VSL) and average path velocity (VAP) of the spermatozoa were significantly decreased (P < 0.001), but amplitude of lateral head displacement (ALH) was insignificant (P > 0.05). When incubated with ROS for 60 minutes, spermatozoa almost lost movement function and all movement parameters of spermatozoa inclined to zero.

CONCLUSIONSWhen normal spermatozoa were incubated with ROS, movement functions of sperm were decreased. It was demonstrated that ROS were one of the causes of the movement dysfunction of spermatozoa.

Adult ; Cell Survival ; drug effects ; Humans ; Male ; Reactive Oxygen Species ; toxicity ; Sperm Motility ; drug effects

OBJECTIVETo investigate the protective effect of histone deacetylase inhibitor NL101 on L-homocysteine (HCA)-induced toxicity in rat neurons, and the toxic effect on normal rat neurons.

METHODSIn the presence of NL101 at various concentrations, HCA (5 mmol/L)-induced changes in cell density, necrosis, and viability were determined in the mixed cultures of rat cortical cells and the primary cultures of rat neurons. The direct effect of NL101 on primary neurons was also observed in the absence of HCA. Histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) was used as the control. After the treatments, cell viability, the density, and morphology of neurons and glial cells, and cell necrosis were determined.

RESULTSIn the mixed cultures of cortical cells, NL101 had no effect on HCA (5 mmol/L)-induced cell number reduction at 0.001-10μmol/L; however, it significantly attenuated necrosis at 1-10 μmol/L, and increased neuronal number at 1 μmol/L. NL101 had no effect on the mixed cortical cells in the absence of HCA. In the primary neurons, NL101 reduced neuronal viability and mildly increased necrosis at 1-10 μmol/L in the absence of HCA, while it significantly attenuated HCA-induced neuronal viability reduction at 0.01-10 μmol/L and reduced neuronal necrosis at 1-10 μmol/L. The effects of NL101 were apparently similar to those of SAHA.

CONCLUSIONNL101 has protective effect on HCA-induced neuronal injury but it is neurotoxic at high concentrations, which is similar to the typical histone deacetylase inhibitor SAHA.

Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Histone Deacetylase Inhibitors ; pharmacology ; Neurons ; drug effects ; Rats

OBJECTIVETo study the safety of Hydrophilic polyacrylamide gel (HPAG) through an animal experiment.

METHODSAfter HPAG was injected underneath the skin of SD rats, tissue specimens were taken for general and histological examinations. The cytotoxicity was evaluated by agar coverage and MTT method.

RESULTSIt was determined that the cytotoxicity was over level-two. The toxicity to kidney was obvious. The local histological reaction was slight and a thin fibrous membrane was formed around HPAG, which became stiff gradually. The shape and location of the injected HPAG was not stable. The HPAG could not be drawn out completely.

CONCLUSIONHPAG has obvious cytotoxicity and is not a suitable material as soft tissue implant for the bad shape and texture.

Acrylic Resins ; toxicity ; Animals ; Cell Survival ; drug effects ; Kidney ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effect of SHENG MAI ZHUSHEYE on the movement parameters and viability of human sperm in vitro.

METHODSWe collected sperm samples from 33 normal fertile men, divided each into two, and cultured them in vitro with SHENG MAI ZHUSHEYE + Hams-F10 and Hams-F10 alone, respectively. Then we measured the straight line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP) and the amplitude of lateral head displacement (ALH) of the sperm by computer-aided semen analysis at 0.5, 1, 2, 4, 8 and 12 h. And the sperm viability was detected.

RESULTSVCL was significantly higher at 8 h (P < 0.05) and very significantly higher at 12 h (P < 0.01) in the SHENG MAI ZHUSHEYE + Hams-F10 group than in the Hams-F10 group. VSL, VAP and ALH were significantly increased in the former group at 4, 8 and 12 h as compared with the latter (P < 0.05). The sperm viability was significantly decreased in the Hams-F10 group at 12 h (P < 0.05).

CONCLUSIONSHENG MAI ZHUSHEYE can improve sperm movement parameters and increase sperm viability in vitro.

Cell Survival ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; In Vitro Techniques ; Male ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects

The effect of trypsin on the separation an subculture of the keratinocytes was investigated in this work. It was found that when 0.25% trypsin was employed for 5 minutes to separate keratinocytes, the number of active keratinocytes and the cells capable of forming colony were higher than those of other experimental conditions. The maximum attached ratio of primary keratinocytes was obtained when skin tissues were treated at 0.05% concentration of trypsin. With the increase of the trypsin concentrations, the attached ratio, attachment rate constant, and colony forming efficiency were all increased. Thus, 0.25% concentration of trypsin was recommended for separating and subculturing the keratinocytes.
Animals ; Cell Count ; methods ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cell Survival ; drug effects ; Keratinocytes ; cytology ; drug effects ; Mice ; Stem Cells ; drug effects ; Trypsin ; pharmacology

OBJECTIVETo investigate the effects of different concentrations of indium chloride (InCl3) on the proliferation of human lung epithelial (Beas-2B) cells and its potential mechanism.

METHODSBeas-2B cells were exposed to different concentrations of InCl3 (0.3, 1.0, 3.0, 10.0, 30.0, 90.0, 270.0, and 810.0 µmol/L) for 24, 48, and 72 h, respectively. The effects of InCl3 on cell proliferation were determined by the CCK-8 assay. The effects of InCl3 on apoptosis were evaluated using annexin V-PI staining followed by flow cytometry. The level of intracellular reactive oxygen species (ROS) in Beas-2B cells after exposure to InCl3 was determined using 2', 7'-dichlorofluorescein diacetate labeling followed by flow cytometry.

RESULTSCompared with the control group, InCl3 at a relatively low concentration (0.3~3.0 µmol/L) significantly promoted cell proliferation (P < 0.05), while InCl3 at a relatively high concentration (30.0~80.0 µmol/L) significantly inhibited cell proliferation after 72 h (P < 0.05). InCl3 at a concentration of 0.3 µmol/L failed to induce apoptosis within 72 h; however, InCl3 at a concentration of 30.0 or 810.0 µmol/L induced substantial early apoptosis after 72 h. Compared with the control group, cells exposed to 0.3 µmol/L InCl3 showed a slight decrease in the level of intracellular ROS within 72 h, while cells exposed to 30.0 or 810.0 µmol/L InCl3 showed a significant increase in the level of intracellular ROS after 72 h (P < 0.05).

CONCLUSIONAt a low concentration, InCl3 stimulates cell proliferation by reducing intracellular ROS. However, at a high concentration, InCl3 inhibits cell viability by elevating intracellular ROS and inducing apoptosis.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Epithelial Cells ; drug effects ; Flow Cytometry ; Humans ; Indium ; toxicity ; Reactive Oxygen Species