With the expanding scale of animal cell culture and increasing demand of biopharmaceutical,the development of serum-free medium based on cell lines and products has become a major task in the field of cell engineer.The statistical methods and tools are popular in the processes of design medium and can be used to evaluate the multiple factors and their interaction scientifically and effectively.The novel microarray and proteomic analysis can improve the performance of identifying the role of medium.The aim is to provide some idea for serum-free medium study through systematically summarizing the newly and commonly used serum-free medium study and design approaches and their characteristics.

ObjectiveTo investigate the analgesic effect of TL-Ⅰ prescription.MethodsHealth ICR mouses were randomly divided into 6 groups: negative control group, positive control group, Tongtian group, large doses group,middle doses group and small doses group. The pain threshold of mouses were detected with hot plate and acetic acid writhing.ResultsTL-Ⅰ prescription can raise the threshold of pain induced by hot plate and reduce the numbers of writhing induced by acetic acid in mice, which was more significant in large doses group. ConclusionTL-Ⅰ prescription can be an effective analgesic.

ObjectiveTo investigate the sedative effect of TL-Ⅰ prescription.Methods72 healthy ICR mice were randomly divided into 6 groups: normal group, model group, positive control group, large doses Chinese materia medica group, middle doses Chinese materia medica group and small doses Chinese materia medica group. The active behavior of migraine-model-mouse was detected with tail suspension test and the analytical system of animal behavior-spontaneous movement.ResultsTL-Ⅰ prescription could prolong the quiescent time of migraine-model-mouse and shorten the active journey and time in the autonomic active box of migraine-model-mouse.ConclusionTL-Ⅰ prescription has sedative effect.

OBJECTIVETo investigate the effects of the static magnetic field (SMF) generated by dental magnetic attachments on osteoblastic proliferations, cell cycle distribution, and apoptosis ratio.

METHODSBy simulating those of the closed-field, the closure process and the open-field Magnedisc 800 magnetic attachments respectively, the in vitro cultured rat osteoblasts were exposed continuously to 12.5, 125, 250 mT SMF. The effects of the SMF on the proliferation of the cells were examined. MTT colorimetry test was performed to detect the effect of the SMF on the vitalities of cells. Flow cytometry was utilized to analyze the cell cycles and cell apoptosis rates.

RESULTSThe SMF exposure didn't change the vital osteoblasts number, the cell cycle distribution and proliferation activities of osteoblasts. The cell apoptosis situation were not observed statistical differences.

CONCLUSIONNo matter the closed-field, the closure process and the open-field magnetic attachments respectively, continuous simulating SMF-stimulation of magnetic attachments couldn't change osteoblasts proliferation activity, cell cycle distribution, and apoptosis ratio.

Animals ; Apoptosis ; Cell Cycle ; Cell Proliferation ; Electromagnetic Fields ; Magnetic Fields ; Magnetic Phenomena ; Magnetics ; Osteoblasts ; Rats

Lack of proper study models has brought difficulties in the study of the mechanism of viral infection, life cycle and pathogenic mechanism of hepatitis C virus (HCV) and also become the major obstacles in development of efficient vaccine and new drugs for hepatitis C. In recent years, the establishment of robust HCV cell culture infection system and HCV transgenic animal provide powerful tools for the analysis of host virus interactions, which facilitate the discovery of antiviral drugs and vaccines for this important human pathogen.
Animals ; Animals, Genetically Modified ; virology ; Disease Models, Animal ; Genotype ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Mice ; Mice, Transgenic ; virology

OBJECTIVETo prepare zirconia toughened nano-composite alumina ceramic powder for dental application. Physical and chemical property of the prepared material were tested, and the effect of development technology on composite powder was also studied in this study.

METHODSNano-composite alumina powder was prepared by surface-induced precipitation method. The effect of pH value and dispersing agent content on volume of alumina suspension sediment was recorded. The effect of ultrasonic time on agglomeration was measured also. X ray diffraction (XRD) was used to analyze powder phase before and after the stabilizer was added. Scanning electronic microscope (SEM) was applied for characterizing the specimen.

RESULTSThe dispersion was better at pH=9 and wt (dispersing agent) = 0.2% approximately 0.3%. Selecting proper ultrasonic time can decrease the agglomeration of powders and lower the average particle size. XRD analysis indicated that the phase composition of the prepared nano-composite ceramic powder was shown as alpha-Al2O3, t-ZrO2 and a small amount of m-ZrO2 after the addition of stabilizer. Through SEM observation, nanometer-sized ZrO2 particles (80 approximately 100 nm) were uniformly located on the surface of submicrometer alumina grains.

CONCLUSIONSBy choosing appropriate preparation method, weakly agglomerated powders with fine particle size can be obtained. The zirconia part of nano-composite powder was transmitted to partially stabled zirconia after the use of stabilizer.

Aluminum Oxide ; chemistry ; Dental Porcelain ; chemistry ; Hydrogen-Ion Concentration ; Powders ; Zirconium ; chemistry

OBJECTIVETo investigate the effects of hepatitis C virus (HCV) on transcription regulation genes of host cells by gene chip assays in cultured cells with intact HCV genome.

METHODSHuh-7 hepatoma cells were cultured and infected with in vitro constructed HCV. The total RNAs, proteins and cell culture supernatants of HCV infected cells and control cells were isolated. Proteins and cell culture supernatants were used to detect the HCV replication and protein expression in cell culture system. The HCV protein expression was detected with Western blotting. Released HCV from infected cells was analyzed by real-time fluorescence quantitative PCR. Total RNA was qualified using 10 g/L agarose gel electrophoresis. cRNA was synthesized, fluorescence labeled and purified, then hybridized with Agilent oligo microarray (20173 probes). Differential expression of genes related to transcription in cell culture system was analyzed.

RESULTHCV was positive in cell culture supernatants and HCV protein expression was also positive according to Western blotting results. Eleven up-regulated and 11 down-regulated genes related to transcription were found after Agilent gene chip screening.

CONCLUSIONIntact hepatitis C virus cell culture system provides an useful tool for study on the affects of HCV infection on transcription regulation genes in host cells.

Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genome, Viral ; Hepacivirus ; genetics ; growth & development ; Hepatocytes ; virology ; Humans ; Liver Neoplasms ; genetics ; pathology ; virology ; Transcription, Genetic

OBJECTIVETo investigate effects of the static magnetic field (SMF) generated by dental magnetic attachment on osteoblastic morphology and surface ultrastructure.

METHODSThe in vitro cultured rat osteoblasts were exposed continuously to 12.5 mT, 125 mT, and 250 mT static magnetic fields for 1, 3, 5, and 7 days. After exposed in SMF, osteoblasts were observed under a phase contrast microscope, and then HE stained and observed under a light microscope. In addition, the cells were observed under a scanning electron microscope (SEM).

RESULTSBy continuous exposure, the different intensities of SMF exposure did not change the vital osteoblast growth pattern or distribution. The SEM photos showed that there were certain changes in cellular microstructures for osteoblasts after exposed to 12.5 mT for 5 to 7 days, as well as 125 mT and 250 mT for 3 to 7 days. The more exposure time increased, the more microvesicles on the surfaces of cells were observed.

CONCLUSIONSContinuous SMF-stimulation could not affect the shape, distribution, and growth pattern of osteoblasts. The SMF of magnetic attachments could lead to certain changes in surface ultrastructures of osteoblasts in this study.

Animals ; Cells, Cultured ; Denture Precision Attachment ; Electromagnetic Fields ; Osteoblasts ; radiation effects ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the expression of keratin 8 (K8) in carbon tetrachloride (CCl(4))-induced liver injury of mice.

METHODSForty ICR mice were divided into four groups. CCl(4) 300 microl/kg body weight in olive oil was injected intraperitoneally for 0, 2, 4, 6 weeks in group A, B, C and D, respectively. Mice were sacrificed 3 d after the last injection and then the vital organs were collected and weighed. RT-PCR and immunohistochemistry methods were used to analyze the expression of keratin 8 in the liver.

RESULTSThe ratios of liver and body weight were increased significantly after administration of CCl(4), which were 5.60 %, 6.87%, 7.83 % and 7.76% at 0, 2, 4 and 6 weeks after injection, respectively. The expression of K8 was increased at the 2 w, 4 w and 6 w after CL(4) administration.

CONCLUSIONThe expression of K8 is positively correlated with the liver injury induced by CCl(4). The accumulation of K8 may be involved in the mechanism of liver injury.

Animals ; Carbon Tetrachloride ; Chemical and Drug Induced Liver Injury ; metabolism ; Female ; Keratin-8 ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; RNA, Messenger ; genetics ; metabolism

OBJECTIVETo investigate the effect of different kinds of mechanical stress on the mRNA expression of integrin beta1 subunit in cultured human periodontal ligament fibroblasts (hPDLF).

METHODSTo scalp and remove the periodontal ligament attached to the mid-third part of the fresh root of young premolars extracted for the cause of orthodontics. Cultured hPDLF by the method of digesting by I-type collagenase combining with tissue adhering. Then hPDLF was isolated and purified by cells passage. The sixth passage's cells were selected to be loaded. A new cyclic strain loading apparatus. Forcel four point bending device was used for mechanically loading. Cells were loaded by three levels (1000, 2000, 4000 microstrain) of tensional and compressive forces and collected at different times (0, 0.5, 1, 4, 8, 12 h) course after strain loading. The quantity of integrin beta1 mRNA in every group was analyzed by means of quantitative real-time PCR with the special primers of up- and down-regulated genes.

RESULTSDynamic mechanical forces down-regulated the expression of integrin beta1 subunit mRNA in hPDLF and the difference in groups by different magnitude, different kinds, and different time of mechanical forces loading were statistically significant. The stronger stimulated forces, the more down-regulated expression. Compression down-regulated the expression of integrin beta1 subunit mRNA more than tension did.

CONCLUSIONDynamic mechanical forces could regulate the expression of integrin beta1 subunit mRNA. The difference among all the groups by different magnitudes, different kinds, and different time of mechanical forces loading were statistically significant.

Cells, Cultured ; Fibroblasts ; Humans ; Integrin beta1 ; Periodontal Ligament ; RNA, Messenger ; Stress, Mechanical