Objective To investigate the effect of PBL combined with EBM applied in the teaching of fever of unknown origin. Methods PBL combined with EBM teaching was applied in fever of unknown origin course for 30 clinical medicine specialty(eight years) students of Tongji class of grade 2009(experiment group), while PBL teaching was applied in fever of unknown origin course for 30 clinical medicine specialty (eight years) students of Tongji class of grade 2008 (control group). After teaching, the theory examination for both basic knowledge and case analysis was organized for all students of both groups. At the same time the questionnaire survey was conducted to 30 students of grade 2009 to evaluate the teaching effect. The results were assessed by using SPSS 18.0 statistical software for the T-test of the experimental group and the control group.Inspection level was α=0.05. Results The theory test score of students in the experimental group was (93.5±3.2) point, signifi-cantly higher than that of the students in the control group(84.7±2.8). There was statistically signifi-cant difference between the scores of the two groups of students (P=0.00). Survey results showed 19 students ( 63 . 33%) thought that the development of PBL teaching combined with evidence-based medicine teaching had its necessity, and 16 students(53.33%) thought that the teaching method im-proved their clinical thinking ability of logical reasoning. Conclusion The concept of PBL combined with EBM has achieved significant resultsinthe teaching offever of unknown origin, and it is necessary to carry out this teaching mode in medical colleges with certain teaching strength.

The objective of this study was to screen out the effective shRNA which can inhibit the gene expression of tumour necrosis factor-alpha (TNF-alpha), to construct the recombinant plasmid and to determine its sequence so as to provide the new approach for searching gene therapy of TNF-alpha related diseases. The primary macrophages were added into 15% DMEM, then cells were adjusted as 2 x 10(7) cells/L and were inoculated in 6-well plate with 3 ml/well, and were cultured at 37 degrees C in a fully humidified atmosphere with 5% CO(2). Cells were stimulated with lipopolysaccharide (LPS) and the concentration of TNF-alpha in the supernatant at different time points was determined by enzyme-linked immunosorbent assay (ELISA). The 5 synthesized DNA sequences which can be transcripted into shRNA were transfected into cells with lipofectamine 2000, then the cells were stimulated with LPS for 24 hours. The concentration of TNF-alpha in the supernatant and the expression of TNF-alpha mRNA were determined by ELISA and reverse transcription polymerase chain reaction (RT-PCR) respectively. The most effective shRNA was inserted into plasmid, and the recombinant plasmid was identified by sequence analysis. The results showed that the concentration of TNF-alpha in the supernatant reached peak after the stimulation with LPS for 24 hours. In the RNA interference group, the shRNA 1 was the most effective one, which could inhibit the expression of TNF-alpha by 59.46% and the expression of TNF-alpha mRNA by 61.2%. The recombinant plasmid was cloned and the sequence of interest was obtained. In conclusion, the most effective shRNA targeting TNF-alpha was successfully screened out and the recombinant plasmid was constructed. The recombinant plasmid may be helpful to search new gene therapy for TNF-alpha related disease.
Animals ; Cells, Cultured ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Macrophages ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Recombination, Genetic ; Transfection ; Tumor Necrosis Factor-alpha ; genetics

The objective of this study was to investigate the function and mechanism of hyperbaric oxygen (HBO) in antagonizing acute graft-versus-host disease (aGVHD) and improving the rate of survival. The lethally irradiated C57BL/6 recipients were injected with bone marrow and lymphocyte of spleen from BALB/c donors and were treated with HBO, cyclosporine A (CsA) and methotrexate (MTX). T lymphocytes and subsets, adhesion molecules and cytokines were detected by flow cytometry, ELISA and RT-PCR respectively. The results showed that the survival rate in HBO group was much higher than that in allogenetic bone marrow transplantation (allo-BMT) group and CsA + MTX group; the numbers of CD3(+), CD4(+), CD8(+), CD4(+)CD11a(+), CD4(+)CD18(+), CD8(+)CD11a(+), CD8(+)CD18(+) lymphocytes in spleen were decreased markedly by HBO and CsA + MTX (p < 0.05); the levels of IL-2 and TNFalpha mRNA and their serum concentrations in HBO group were much lower than those in allo-BMT group but were higher than those in CsA + MTX group; the levels of IL-4 and IL-10 mRNA in HBO group were much higher than those in allo-BMT group and CsA + MTX group. It is concluded that HBO has more remarkable advantage in improving the rate of survival than CsA + MTX, its mechanism of anti-aGVHD is tightly correlated with the transform of T cell and its subsets and the expression of adhesion molecules and cytokines.
Acute Disease ; Animals ; Bone Marrow Transplantation ; adverse effects ; Cytokines ; biosynthesis ; Female ; Graft vs Host Disease ; etiology ; therapy ; Hyperbaric Oxygenation ; Lymphocyte Transfusion ; adverse effects ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes ; immunology ; Whole-Body Irradiation

The aim of this study was to explore the effect of hyperbaric oxygen (HBO) preconditioning on the rejection of skin allograft in mice and its molecular mechanism. BALB/c donor mice and C57BL/6 recipients received hyperbaric oxygen preconditioning once a day for 7 days. After skin transplantation, the recipients were treated with cyclosporine A (CsA) intraperitoneally. Immunofluorescent staining technique and flow cytometry were used to observe the influence HBO on percentage of spleen lymphocytes CD3+, CD4+, CD8+ and cell adhesion molecule LFA-1 (CD11a/CD18). The results showed that as compared with control, the numbers of CD3+, CD4+, CD8+, CD4+CD11a+, CD4+ CD18+, CD8+CD11a+, CD8+CD18+ lymphocytes of spleen decreased in HBO preconditioning groups and CsA group, and decreased markedly in HBO preconditioning combined with CsA group (p<0.05); the general state of recipient mice in HBO preconditioning combined with CsA group was better than that of recipient mice received HBO preconditioning or CsA only. It is concluded that the method of HBO preconditioning combined with traditional immunosuppressive agent CsA has remarkable advantage in inhibiting the rejection of skin graft. Its molecular protective mechanism is correlated with the expression of adhesive molecules on T cell subsets.
Animals ; Cell Adhesion ; Cell Adhesion Molecules ; pharmacology ; Cyclosporine ; pharmacology ; Female ; Graft Rejection ; prevention & control ; Graft Survival ; Hyperbaric Oxygenation ; Lymphocyte Count ; Lymphocytes ; cytology ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Skin Transplantation ; immunology ; Spleen ; cytology ; Transplantation Conditioning ; methods

Adenocarcinoma ; metabolism ; pathology ; Carcinoid Tumor ; metabolism ; pathology ; surgery ; Child ; Chromogranin A ; metabolism ; Common Bile Duct ; pathology ; surgery ; Common Bile Duct Neoplasms ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Duodenum ; pathology ; surgery ; Gallbladder ; pathology ; surgery ; Humans ; Keratins ; metabolism ; Lymphoma ; metabolism ; pathology ; Male ; Mucin-1 ; metabolism ; Neoplasm Invasiveness ; Rhabdomyosarcoma ; metabolism ; pathology ; Stomach ; pathology ; surgery ; Synaptophysin ; metabolism

Humans ; Infant ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Mucin-1 ; metabolism ; Nephrectomy ; Precancerous Conditions ; metabolism ; pathology ; surgery ; Vimentin ; metabolism ; WT1 Proteins ; metabolism ; Wilms Tumor ; metabolism ; pathology ; surgery

Child ; Child, Preschool ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Infant ; Male ; Wilms Tumor ; genetics ; metabolism ; Wnt Proteins ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

Adrenal Gland Neoplasms ; pathology ; surgery ; Adrenalectomy ; Child ; Diagnosis, Differential ; Follow-Up Studies ; Ganglioneuroblastoma ; pathology ; Hematopoiesis, Extramedullary ; Hodgkin Disease ; pathology ; Humans ; Leukemia ; pathology ; Male

Objective To investigate the diagnostic value of twinkling artifact in urinary calcium stones.Methods Calcium oxalate monohydrate,hydroxyapatite and whitlockite stone phantoms were prepared and embedded in porcine kidneys for ultrasound scanning.The length and width of twinkling artifact were measured.The intensity of twinkling artifact and the contrast-to-noise ratio (C/N ratio) of acoustic shadowing were recorded and all the data was analyzed statistically.Results All the stone phantoms generated twinkling artifact and acoustic shadowing.The difference of the intensity,length and width of twinkling artifact and the C/N ratio of acoustic shadowing had statistically significant each composition (P ＜0.01).There was no overlap between 95％ confidence interval of the intensity of twinkling artifact for any two compositions,and any two compositions of stones could be differentiated by the intensity of twinkling artifact (P ＜0.05).The C/N ratio of acoustic shadowing of calcium oxalate monohydrate and whitlockite stone phantoms were higher than those of hydroxyapatite ones (P ＜0.05).The receiver operating characteristic (ROC) curve displayed that the cutoff value of 1 127 of the intensity of twinkling artifact could be used to diagnose hydroxyapatite stones.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and area under the curve (AUC) were 74 ％,68 ％,69.8％,72.3％ and 0.743 respectively.Conclusions Twinkling artifact could be used to differentiate the calcium stones,which will be helpful for clinical treatment.

Objective To quantitatively analyze the value of color Doppler twinkling artifact and CT in distinguishing urinary stone composition of uric acid from that of cystine,and to assess the ability of color Doppler twinkling artifact and CT to diagnose urinary stone composition of uric acid.Methods Artificial stones made of uric acid and cystihe were inserted in porcine kidneys respectively.Color Doppler ultrasound and CT scanning were performed in vitro and twinkling artifact videos were recorded.The CT attenuation value,the width and length of twinkling artifact (TAW,TAL) were measured.The color pixels representing twinkling artifact intensity (TAI) were calculated.The differences between artificial cystine and artificial uric acid stones in TAW,TAL,TAI and CT attenuation value were compared.The receiver operating characteristic (ROC) curve was used to assess the ability of TAW,TAL,TAI and CT attenuation value to diagnose urinary stone composition of uric acid.Results The TAW [(4.16 ± 1.54)mm vs (4.97 ± 1.93)mm],TAL[(3.75 ± 1.05)mm vs (5.12 ± 2.51)mm] and TAI [(3 005.50 ± 812.33) vs (5 433.86±2 505.81)] of artificial cystine stones were less than those of artificial uric acid stones while the CT attenuation value of artificial cystine stones [(573.75 ± 110.10)HU vs (364.09 ± 67.28)HU] were greater (P ＜0.05).The ROC curve was displayed to diagnose urinary stone composition of uric acid,and the area under curve of CT attenuation value was bigger than that of TAI (0.936 vs 0.817,Z =2.308,P =0.021).Combination of color Doppler twinkling artifact and CT value would dramatically increase the diagnostic accuracy to 98％.Conclusions Both color Doppler ultrasound and CT were reliable examinations for distinguishing urinary stone composition of cystine from that of uric acid.Combination of color Doppler ultrasound and CT would be good references to diagnose urinary stone composition of uric acid.