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 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

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

Objective To explore the bioeffect of different parameters on 4 cell lines by ultrasoundmediated microbubble destruction.Methods The orthogonal experimental design was used to investigate the effect of three factors on the bioeffects of four cell lines under three levels.Three factors included microbubble concentration,sound intensity,irradiation time.Human breast tumor (MCF-7) cells,ovarian tumor (A2780) cells,liver tumor (Bel7402) cells and thyroid tumor (ARO) cells were exposed to ultrasound in the presence of SonoVue.The cell survival rate was determined by MTT methods and the cell luminosity factor was detected by flow cytometry.Results The optimum parameters for Bel7402 and ARO cell were the same (A2B3C2),and they were different from those from MCF-7 (A3B1C1) and A2780 (A1B3C3) cell.The cell survival rates for 4 cell lines were above 75％,and the cell luminosity factors were different among 4 cell lines.Conclusions The optimum parameters by ultrasound-mediated microbubble destruction for different cell lines are different,and under the optimum parameters the bioeffects of different cell lines are different.

Background Retinal development continues during the early postnatal period in mammals.Correct arrangement of layers and precise location of various cells in the retina are vital for forming normal visual function during critical period plasticity.Spectral-domain optical coherence tomography(SD-OCT)provides highquality in vivo retinal imaging and the possibility to measure retinal thickness longitudinally. Objective The present study was to investigate the changes of retinal thickness during critical period plasticity in rats. Methods In vivo consecutive scanning of retinal image was performed in 10 SPF Sprague-Dawley rats at postnatal day 14(P14),P18,P21,P24 and P42 with SD-OCT,and retinal histopathological examination was used to detect retinal morphologic changes at the same postnatal ages in 20 matched rats.The whole retinal thickness,the thickness from inner limiting membrane(ILM)to inner plexiform layer(IPL),the thickness of inner nuclear layer(INL)and the thickness from outer nuclear layer(ONL)to retinal pigment epithelium(RPE)were measured using Cirrus HD-OCT system and HMIAS-2000 Imaging System in retinal sections.The measurement parameters by Cirrus HD-OCT and those by hematoxylin-eosin staining were compared.The use of animals followed the Statement of National Institute of Health (USA). Results In vivo high-resolution images of rat retinas with SD-OCT compared well with histology,which enabled quantitative comparison of the SD-OCT and histological data during critical period plasticity in rats.From P14 to P42,the retinal thickness gradually decreased with the increase of rat ages(F=15.425,P=0.000),and so were the thickness from ILM to IPL,the thickness of INL and the thickness from ONL to RPE(F=3.973,P=0.007;F=17.529,P=0.000;F=7.038,P=0.000).The retinal thickness,thickness of INL.thickness from ONL to RPE measured by Cirrus HD-OCT were significantly correlated with those measured by retinal sections among P14,P18,P21,P24 and P42 rats(r=0.794,P=0.000;r=0.784,P=0.000;r=0.681,P=0.000). Conclusion SD-OCT is a demonstratably valuable technology to study the structure of retinas in rats.The retinal thickness is shown to reduce in thickness throughout the development of the retina during critical period plasticity due to the decrease in thickness of INL and the distance from the ONL to RPE,as illustrated by OCT scanning.

BackgroundMonocyte chemotactic protein-1 (MCP-1)plays an important role in the tumor,inflammation,diabetic retinopathy and other neovascular disease,but the expression and the role of MCP-1 in the oxygen induced retinopathy(OIR) model have rarely been reported. Objective This study was to investigate the expression of MCP-1 in the retina development of newborn mouse and in mouse models with OIR.Methods C57BL/6J newborn mice were divided into two groups and 60 mice in each group.Mice in OIR group were exposed to 75％ oxygen for 5 days and then to room air.All mice in normal control group exposed to room air only.Ten mice in each group were randomly chosen and sacrificed at postnatal 5,7,12,14,17,21 days.The expression of MCP-1 in mouse retina was detected with the method of immunohistoehemistry and reverse transcription polymerase chain reaction(RT-PCR).Results MCP-1 positive cells were seen in normal mouse retina.Up-regulation of MCP-1 positive cells was detected both in 12 days in normal control group and in 14 days in OIR group.MCP-1 mRNA was detected in mouse retina at 5 days,and a transient up-regulation of MCP-1 mRNA was observed in 12 days in normal control group.MCP-1 mRNA in OIR group significantly increased in 14 days in comparison with the normal control group( P =0.028,P =0.001 ). Conclusions Expression of MCP-1 is detectable in whole retinal development procession of mice.A transient up-regulation of MCP-1 expression is detected in the critical period of retinal vascular development in mice models with OIR,which is closely related to the retinal vascular development and progression of retinal new vessels.

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.