To train self-learning ability,clinical reasoning and comprehensive analytical ability of clinical medical postgraduates,we introduced the classic cases from the New England Journal of Medicine (NEJM) and made the coherent learning method of Learn classic cases-report clinical cases-write case reports.Students were required to learn the classic cases of NEJM independently,to report clinical cases based on classic cases' level and to write a high level case report.We encouraged students to learn independently and to learn to use it.The practice has proved that the coherent learning method is a gradual and systematic learning method,which can train clinical medical postgraduates' all-round abilities and help them to grow up to qualified doctors.

Objective To study the expression of HOXB9 gene in Hela cells,Mocoy cells, SP2/0 cells and U251 cells. Methods The expression of HOXB9 gene was detected by semi-quantitative RT-PCR method. Results Hela cell and U251 cell expressed HOXB9 gene, which SP2/0 cell and Mocoy cell didn't express it. Conclusion The expression of HOXB9 gene was different in different cells.

Objective:To investigate the effect of glucocorticoid on treating class IV lupus nephritis(LN) by comparing the content of serum transforming growth factor beta(TGF ? 1).Methods:The study consisted of 33 objects,15 health controls as well as 18 cases of IV LN.The level of serum TGF ? 1 was analyzed by ELISA.Results:The level of serum TGF ? 1 was greatly reduced after the patients were treated with glucorcorticoid.Conclusion:Timely and sufficient glucocorticoid may prevent IV LN from developing into glomerulosclerosis.

Objective:Observing the mechanism of apoptosis in the carcinoma of urinary bladder BIU 87 cell strain induced by staphylococcus enterotoxin A (SEA).Methods:The multiplicative inhibitory rate of the BIU 87 cell strain and cell cycle affected by SEA was tested with MTT colorimetry and low cytometry.Results:SEA restrain obviously the BIU 87 cell strain and have dosage dependence and obstruct the cell multiplication processes from G 1 period to S period.A crowd of cells was seen G 1 period.Conclusion:SEA having effect of cytotoxicity can make BIU 87 cell strain apoptosis and prevent the metastsis of the carcinoma of urinary bladder by pouring it into bladder.

Objective To describe the status quo of Aged care willingness among nursing undergraduates,and to explore its influencing factors. Methods A total of 382 nursing undergraduates were enrolled by convenience sampling method. They were investigated with a self-designed questionnaire about the demographic data, the Kogan′s Attitudes toward Old People Scale, the Palmore′s Facts on Aging Quiz Scale and College Students′ Gratitude Trait Questionnaire. Results The overall aged care willingness score for nursing undergraduates was 17.75 ± 3.62. Appreciation, old training, happiness, retribution and living with the older people entered the multiple regression equation. Conclusions The Aged care willingness level of 382 nursing undergraduates is at medium level. Appreciation, old training, happiness, retribution and living with the older people are important influencing factors.

Objective:To produce and identify antiCK20 monoclonal antibody.Methods:Lymphocytes from the spleen of mice being immunonized by CK20 antigen were fused with the myeloma cell line(SP2/0) using PEG4000.Hybridodma cells were established by selective growth of the fusion cells in the HT medium,and the presence of antiCK20 antibody was screened by inderect ELISA,and the clonality was achieved by limiting dilution.We have incubated cloning cells into mouse abdominal cavity to produce ascitic fluid contained monoclonal antibody.Chromatography with SPA-Sepharose CL-4B affinity column were emploied to isolate the monoclonal antibody from ascitic fluid.Finally,the antibody were tested the activities and sentivities,isoforms and titer through Western blot,two directions immuning diffusion of agar and ELISA.Results:Only one hybridoma cell line,secreating McAb against CK20,had been established.The modal number of chromosome is 101(99-103).The results of identifications showed that the antibodies kept high activitis and sensitivitis in detecting sample.The titer of ascitic fluid and the McAb purified are 1∶10~6 equally.The immunoglobulin of the McAb is classified as IgG1.Conclusion:AntiCK20 monoclonal antibody have been produced succesfully with high sensitive and active and was named L20031030.

Objective To investigate the release and intracellular localization of high mobility group box chromosomal protein 1(HMGBl)in the peripheral blood monocytes of rheumatoid arthritis(RA) patients and the inhibitive effect of thaiidomide.Methods 19 RA patients and 20 healthy controls were included in the study.Monocytes were separated from peripheral blood with Ficoll density gradient centrifugation.Monocytes were treated with 100 ng/ml tumor necrosis factor α(TNFa)or 100 ng/ml TNFα plus 40 μg/ml thalidomide and grown in an incubator at 37℃ with 5%CO,for 24 hours.The cuIture supernatants of the monocytes were collected.HMGB1 level in the culture medium was detected with Western blot.In addition,the intraceUular localization of HMGB1 in the fflonocytes was investigated with immunocytochemical analysis. Results Without stimulation. the release of HMGBl protein was significantly increased in the culture supernatants of peripheral blood monocytes from RA patients as compared with that from healthy controls(P<0.05).TNFα(100 ng/ml)did not further increase the release of HMGBl in the monocytes from the patients with RA.Thalidomide(40 μg/ml)could inhibit the release of HMGB1 in the monocytes from RA patients stimulated with TNFα(P<0.05).In the monocytes from RA patients,HMGBl was mainly localized in the nucleus.Treatment with TNFOL(100 ng/ml)for 24 hour resulted in a cytoplasmic translocation of HMGB1,which was inhibited significantly by thalidomide. Conclusion TNFα induces the release and cytoplasmic translocation of HMGBI in the peTipheral blood monocytes of RA patients and thalidomide inhibits the release and translocation of HMGB1.

Humans ; Receptor, ErbB-2 ; Staining and Labeling

BACKGROUND:Notch singling pathway is very important for cellproliferation and differentiation, but its role is stil unknown during chondrogenesis of human umbilical cord mesenchymal stem cells. OBJECTIVE:To investigate the effect of N-[N-(3,5-difluorophenacetyl-L-alanyl)]-(S)-phenylglycinet-butyl ester (DAPT) on inducing human umbilical cord mesenchymal stem celldifferentiation into chondrocytes. METHODS:Human umbilical cord mesenchymal stem cells were isolated from human umbilical cord, then were induced to differentiate into chondrocytes. There were four experimental groups:non-induced group, high-glucose Dulbecco’s modified Eagle’s medium containing 5%fetal bovine serum and 1%double antibody;induced group, induced medium containing 6.25 mg/L insulin, 6.25 mg/L transferrin, 10μg/L transforming growth factor beta 1, 0.1μmol/L dexamethasone, 50 mg/L vitamin C, 5%fetal bovine serum and 1%double antibody;dimethyl sulfoxide group, induced medium containing 0.1%dimethyl sulfoxide;DAPT group, induced medium containing 5μmol/L DAPT. RESULTS AND CONCLUSION:After chondrogenic induction, the morphology of human umbilical cord mesenchymal stem cells became polygon and positive for toluidine blue and immunofluorescence staining;the expression of Jag-1, PS-1, Notch-1 and Hes-1 decreased significantly (P<0.01). After the addition of DAPT, compared with the induced group, the relative gene expression of Jag-1, PS-1 and Hes-1 decreased markedly (P<0.01), the relative gene expression of Notch-1 decreased obviously as wel (P<0.05), and the contents of proteoglycan and col agen type II proteins decreased significantly (P<0.01). At the same time, the relative gene expression of proteoglycan decreased obviously (P<0.05), and the relative gene expression of col agen type II decreased in part. Notch signaling pathway exists in human umbilical cord mesenchymal stem cells, once chondrogenesis begins, the signaling strength wil decline rapidly. DAPT may prevent human umbilical cord mesenchymal stem cells from differentiating into chondrocytes by Jag-1-Notch-1-Hes-1 pathway.

This study is to report the study of protective effects of myricitrin against oxidative stress-induced apoptosis of vascular endothelial cells and the investigation of the possible mechanisms of action of myricitrin. The model of H2O2-induced apoptosis of vascular endothelial cells was used to determine the protective effects of myricitrin. The levels of LDH, MDA and the activities of SOD, NO were measured using the respective kits. The H2O2-induced apoptosis of vascular endothelial cells was detected using MTT reduction, TUNEL assay, JC-1 and ROS staining. The activation of Caspase-3 was also measured by fluorometry. The expression of apoptosis-related proteins was determined with Western blotting assay. Myricitrin had significant protective effects against H2O2-induced apoptosis of vascular endothelial cells in a time- and dose-dependent manner. The results show that myricitrin could attenuate H2O2-induced decrease in the activities of SOD (P < 0.01). Myricitrin could decrease the levels of LDH, MDA and increase cell viability and the mitochondrial membrane potential (P < 0.01). Myricitrin had protective effects in a dose-dependent manner between 32 micromol x L(-1) to 64 micromol x L(-1). Myricitrin pretreatment could attenuate H2O2-induced increase of p-ERK. Moreover, myricitrin pretreatment could up-regulate the expression of the anti-apoptotic protein Bcl-2, down-regulate the expression of the pro-apoptotic protein Bax, and decrease the expression of Caspase-3, 9. In conclusion, myricitrin had significant protective effects against H2O2-induced apoptosis of vascular endothelial cells. Myricitrin can enhance the activities of anti-oxidative enzymes and decrease the production of free radicals. The possible mechanisms of action of myricitrin are due to myricitrin-mediated inhibition of phosphorylation of the apoptosis signaling pathways-related kinase ERK, up-regulation of the expression of the anti-apoptotic protein, and down-regulation of the expression of the pro-apoptotic protein.