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 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 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 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.

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.

Humans ; Receptor, ErbB-2 ; Staining and Labeling

Objective To investigate the effect of moderate pain on attentional bias towards emotional pictures among healthy subjects.Methods Thirty-two healthy college students aged from 17 to 26 (21.8±2.2;16 males and 16 females) participated in this study.A tourniquet was tied to each subject's left upper arm 1 to 2cm above the cubits horizontal grain.Pain was inflicted by inflating the tourniquet,and the pressure was maintained at 26.66kPa.While tourniquet was inflated (with pain) or not (no pain),each subject was asked to finish a pictorial dot-probe task with three kinds of pictures-emotionally positive,negative and neutral.In experiment 1,subjects performed the dot-probe tasks with the contralateral hand while the tourniquet was tied on the left upper arm without inflation.In experiment 2 the tourniquet was inflated until the subject completed the dot-probe task (for about 10min).The reaction times (RTs) and the error rates (Ers) in the recognition task were recorded,and the intensity of the subject's pain and discomfort were measured using a verbal rating scale.Results The subjects reported moderate to severe pain with the tourniquet inflated.The RT and ER data were analyzed using two-way analysis of variance (ANOVA) which showed a significant difference between the average RTs of the males (482±73ms without pain and 466±82ms with pain) and those of the females (536±90ms without pain and 519±100 ms with pain).The average ER was significantly different between the pain (2.38±1.49)％ and no pain (1.09±0.82)％ conditions in both groups.Holn-Sidak multiple comparison testing showed significant differences in both groups' average ER between the negative picture (3.81±1.73)％ and the positive picture (1.66±0.97)％,and between the negative and neutral pictures (1.68±0.8) ％ in the pain condition.Mild attentional avoidance was observed with the positive [pain condition (-5.1±4.8) ms and no pain (-4.6±4)ms] and negative pictures [pain condition (-3.43±6) ms and no pain (-0.79±4.1)ms],but no significant difference was found between the pain and no pain conditions.Conclusion The error rate in a pictorial dot-probe task is influenced by pain,especially with negative pictures.

Objective:To design and construct the replication-deficient recombinant adenovirus Ad-siCTGF which can silence the expression of connective tissue growth factor (CTGF) by RNA interference and verified its function. Methods:A specific sequence, which was verified to be able to silence CTGF gene with high efficiency, was cloned into pSES-HUS vector to produce the shuttle plasmid pSES-siCTGF. The plasmid after Pme Ⅰ linearization was cotransduced with pAdEasy into BJ5183 E.coli strains to construct recombinant vector Ad-siCTGF. After linearization treatment with Pac Ⅰ enzyme digestion Ad-siCTGF was transfected into HEK293 cells via liposome mediation. The recombinant adenovirus was packaged. The titer of the Ad-siCTGF was increased after three times of cross-infection. 4T1 cells were infected with the adenovirus. The silencing efficiency was tested by real-time fluorescence quantitative (RFQ)-PCR and Western blotting.Results:Pac Ⅰ enzyme digestion electrophoresis indentified that recombinant adenovirus was successfully constructed. The titer of the recombinant adenovirus Ad-siCTGF was 2.6×10~(10) pfu/mL after amplification and purification. The CTGF mRNA and protein expression in 4T1 cells were decreased by 36.27% and 31.56%, respectively, compared with the control groups.Conclusion:The recombinant adenovirus which can silence the expression of CTGF was successfully constructed. It laid a good foundation for further investigation of the action mechanism of CTGF in tumor cells.