Objective To demonstrate that miR-483-5p and its targeted gene ERK1 involved in the regulation of proliferation and apoptosis of human granulosa cells.Methods ① The ERK1 mRNA and protein level expression were detected by PCR and Western blot after miR-483-5p overexpression in vitro normal human granular cells.De-tected the relative luciferase density after cotransfecting miR-483-5p mimics and its control respectively with wild or mutant ERK1 mRNA 3`UTR cloned luciferase report vector in granular cells.② Granular cells proliferation and ap-optosis were detected by MTT and TUNEL after transfecting miR-483-5p mimics or ERK1 siRNAs alone or simulta-neously.Results ① It showed that both ERK1 mRNA and protein in granular cells were markedly downregulated after the transfection of miR-483-5p mimics.A significant relative luciferase activity decrease were detected in the granular cells co-transfected with ERK1 wild and miR-483-5p mimics comparison with the control mimics,but not in the cells co-transfected with the ERK1 mutant and miR-483-5p mimics.② When miR-483-5p mimics and ERK1 siRNAs were alone or co-transfected into granular cells,the proliferation was inhibited while apoptosis trend was monitored to be promoted in all cases.No obvious statistic difference was shown between each other.Conclusion MiR-483-5p is involved in the regulation of the proliferation and apoptosis of human granulosa cells by directly binding to the target gene ERK1.

Objective The aim of this research was to provide a basis for the development of preventive and curative measures against the toxoplasma infection by investigating the current infection of toxoplasma and its epidemic characteristics among the population of the Cangzhou city.Methods Antibodies IgG and IgM against toxoplasma in the sera were determined by ELISA during Jan.-May,2007.Results Six hundred and thirty-servn subjects were screened.The positive rate was 6.3% for total antibodies,5.2% for IgG and 1.1% for IgM.The infection rate was higher in the population aged 19-24 years and 49-60 years.The positive rate was higher in farmers living in countryside,those who were poor educated and who had bad habit of cooking food without separating knives and chopping boards to handle raw food and ready-to-eat food and washing their hands after cutting raw meat(P

Objective To establish a rapid test method for the determination of five sweeteners and two preservatives in the milk beverage by HPLC.Methods Trough a set of pretreatment such as dilution,protein precipitation,high-speed centrifugation,the samples were tested.With NUCLEODUR C18 RP-column separation,phosphate buffer(pH=5.0) and acetonitrile were used as the mobile phase,gradient elution analysis was conducted,the detective wavelength was 205nm.Results The separation of one sample was achieved within 15 min,five sweeteners and two preservatives could be well separated,the linear range was 5.00-100.0 mg/L,r≥0.9993,RSD was 1.5%-4.2%(n=6),the average recovery rate was 85.2%-108%.Conclusion This method is simple and quick with better reproducibility and selectivity,it is an effective way to determine sweeteners and preservatives in milk beverage.

Practical teaching of medical ethics is the key point in judging the effectiveness of the course.After 27 years of teaching efforts and exploration,the teaching of medical ethics in our college has gradually formed the basic form of practical teaching,such as case teaching,group discussion,scientific research activities,case assessment,the combination of medical ethics teachers and clinical teachers in the teaching process.But more attention and efforts are still needed in the following issues,including building a scientific evaluation system for the course of medical ethics,focusing on the continuity of practical and systematic teaching,dedicated practice of medical ethics course,and handling new arising problems in modern medical and healthcare practice.

Cellular senescence is one of the important steps against tumor. This study was to observe the characteristics of boningmycin induced senescence of human tumor cells. MIT method and clone formation assay were used to detect the growth-inhibitory effect. Cellular senescence was detected with senescence-associated beta-galactosidase staining. Cell cycle distribution and accumulation of intracellular reactive oxygen species (ROS) were analyzed with flow cytometry. Protein expression was detected by Western blotting. The results showed that the growth-inhibitory effect of boningmycin was obviously stronger on human oral epithelial carcinoma KB cells than that on non-small cell lung cancer A549 cells. Comparison to the similar action of doxorubicin, that boningmycin induced the features of cellular senescence in both cell lines, its due to the arrest at G2/M phase and an increase of ROS level. The molecular senescence marker P21 increased significantly after boningmycin treatment at a dosage of 0.1 micromol x L(-1), whereas a higher concentration of it induced apoptosis. The results indicated that cellular senescence induced by boningmycin was one of its mechanisms in tumor suppression.

It is an essential educational goal to strength experimental teaching and improving the practical skill of medical postgraduate students in medical colleges and universities.We made anexploration and practice on how to improve the practical skill and innovative ability of postgraduate students in medical immunology experimental teaching.The article points out that optimizing the design of experimental course,innovating the teaching methods,standard appraised of experiment and improving the quality of teacher are the efficient ways to enhance the practical skill of postgraduate students.

BACKGROUND:Damage to nasal ciliated epithelial cels can lead to a severe injury in nasal biological function. Compared with other adult stem cels, human umbilical cord blood stem cels have better differentiation potential.
OBJECTIVE: To explore the feasibility of human umbilical cord blood stem cels differentiating into nasal ciliated epithelial cels through in vitro culture and induction techniques.
METHODS:Normal and healthy umbilical cord blood samples were colected to isolate human umbilical cord blood stem cels, folowed by identification and subculturein vitro. Umbilical cord blood stem cels at passage 3 were infected with recombinant adeno-associated virus carrying enhanced green fluorescent protein and cultured using air liquid interface culture method. Thereafter, PCR assay was employed for detecting MUCS expression in cultured stem cels at 1 and 2 weeks after induction, and immunofluorescent staining for FOXJ1 was performed at 3 weeks.
RESULTS AND CONCLUSION:After subculture, passage 3 umbilical cord blood stem cels that could express stem cel surface markers were visible in a uniform shape and had good refraction. After 3 hours of gene transfection, green fluorescence issued from the passage 3 cels were visible, and the cel positive rate was up to 96.2% until 48 hours, indicating good transfection efficiency. RT-PCR findings showed that MUC8 mRNA had no expression in the umbilical cord blood stem cels, but expressed strongly in the nasal ciliated epithelial cels, whose expression was weak at 1 week of culture and increased at 2 weeks. Additionaly, the positive expression of FOXJ1 red fluorescence was observed under the transfection of green fluorescent protein. These results suggest that human umbilical cord blood stem cels could differentiate into nasal epithelial cels under suitable conditions.

Systemic inflammatory response syndrome (SIRS) is a pathophysiological process of critical children.It's pathogenesis involved many sides such as immune dysfunction,gene expression regulation and intestinal barrier.By studying the regulation mechanism of SIRS,early diagnosis of SIRS can be made.According to the inflammatory status and each sides of pathogenesis,effective control results can be taken in time to stop the further developmentof SIRS and improve outcomes of critical children.

Objectives To investigate the effect of matrine (MAT) on adriamycin (ADR) induced podocyte injury in vitro and explore the function of the mammalian target of rapamycin (mTOR) protein signaling pathway during the intervention. Methods ADR (1μmol/L) was used to induce the model of podocyte injury and then the podocytes were intervened by 10, 20 and 40μg/ml MAT for 24 hours. The viability and apoptosis rate of podocytes, mRNA and protein expressions of desmin and mTOR were detected. The effect of different concentrations of MAT on ADR-induced podocytes was analyzed. Results Com-pared with the control group, declined viability of podocytes (P<0.001), increased percentage of apoptotic podocytes (P<0.001), and increased expression of damage marker desmin (P=0.002) were observed in ADR group. In ADR group, after intervention with MAT of 10-40μg/ml, increased viability of podocytes (P<0.05), decreased percentage of apoptotic podocytes (P<0.05), and decreased expression of damage marker Desmin were found (P<0.05). The protein expression of mTOR and phosphorylation state of mTOR (p-mTOR) decreased in ADR induced-podocytes (P<0.05), and after intervention with MAT of 10-40μg/ml, the expression of mTOR and p-mTOR increased (P<0.05). The expression of mTOR downstream target protein s6k1 and 4EBP1 mRNA decreased in ADR group (P=0.071), while increased after intervention with MAT of 10-40μg/ml (P<0.05). Conclusions 10-40μg/ml MAT have protective effects on ADR-induced podocytes in vitro. The protection mechanism may be related to mTOR signaling pathway.

Objective To establish the HeLa cell line that can stably express EYFP fluorescent protein as the model for anion channel blocker (halide ion) screening ,which lays the foundation for high throughput screening of anion channel blocker (halide ion) .Methods Through gene recombination technology ,a new lentivirus vector which can express mutant protein YFP (EYFP‐H148Q/I152L) and puromycin resistance ,was built .The mixture of lentivirus vector and packaging plasmid was transfected into 293T cells to produce lentivirus particles . After infection of HeLa cells by the lentivirus particles ,puromycin was used to screen the cells as YFP‐positive HeLa cell line .Then cell amplification was carried out after purification and efficiency of EYFP‐H148Q/I152L was further detected by Real‐time quantitative PCR (RT‐PCR) and Western blot .We then verified the activity of EYFP‐HeLa transfected cell line as a screening model of anion channel blocker .Results Gene sequencing verified that EYFP‐H148Q/I152L was successfully inserted into lentivirus vectors .RT‐PCR and Western blot results showed that the target gene was overexpressed in HeLa cells . The specific yellow fluorescence of EYFP of HeLa cells could be observed under fluorescence microscope with the efficiency of nearly 100% . I- (low permeability ) solution stimulated the opening of anion (halogen) channels ,and the yellow fluorescence was quenched by I - flow into cells . Conclusion The EYFP‐HeLa cell line can stably express EYFP yellow fluorescent protein and is sensitive to the internal flow of I - .Therefore ,it can be used as an ideal screening model of anion channel blocker (halide ion) .