Objective To verify whether the extracellular domain of flt 1 has anti angionesis activity in vivo. Methods A recombinant plasmid pcDNA 3.1/flt 1 n3 was transfected into the human bladder carcinoma EJ cell by lipofectamin, which was constructed by inserting the N terminal first three IgG like domain of VEGF receptor flt 1 (flt 1 n3 ) into eukaryotic expression vector pcDNA 3.1. Results The rflt 1 was functionally expressed in stably transfected EJ and the product secreted in the medium could be specifically bind to rhVEGF165.The result showed that the bladder cancer transfected with rflt 1 had a lower microvessle density than the control. Conclusions It is proved that the expressed rflt 1 n3 can inhibit tumor growth and angiogenesis in nude mice model.

With the change of medical model and the transfer of health to communities and rural towns,the expanding enrollment of colleges and universities makes collegiate internship progrms more and more difficult Clinical College of Guilin Medical University explores the new methodology of teaching practice,constructs and practises the new molde of internal madichine teaching,Combines the coummunity internship programs with hospital internship programs.and was improved the quality of practice teaching

Objective To evaluate the value of multi-slice CT(MSCT)classification in the assessment of the hilar cholangiocarcinoma resectahility.Methods Thirty patients with surgically and histopathologically proved hilar cholangiocarcinomas who underwent preoperative MSCT and were diagnosed correctly were included in present study.Transverse images and reconstructed MPR images were reviewed for Bismuth-Corlette classification and morphological classification of hilar cholangiocarcinoma.Thcn MSCT classification was compared with findings of surgery and histopathology.Curative resectabilty of different types according to Bismuth-Corlette classification and morphological classification were analyzed with chi-square test.Results In 30 cases,the numbers of Type Ⅰ,Ⅱ,Ⅲa,Ⅲb and Ⅳ according to BismuthCorlette classification were 1,3,4,5 and 17.Seventeen patients underwent curative resections,among which 1,2,1,4 and 9 belonged to Type Ⅰ,Ⅱ,Ⅲa,Ⅲb and Ⅳ respectively.However,there was no significant difference in curative resectability among different types of Bismuth-Corlette classification(X2=0.9875.P＞0.05).In present study,the accuracy of MSCT in Bismuth-Corlette classification reached 86.7%(26/30).The numbers of periducatal infiltrating,mass forming and intraductal growing type were 13,13 and 4,while 6,8 and 3 cases of each type underwent curative resections.There was no significant difference in curative resectability among different types of morphological classification(X2=1.2583,P＞0.05).The accuracy of MSCT in morphological classification was 100%(30/30)in this study group.Conclusion MSCT can make accurate diagnosis of Bismuth-Corlette classification and morphological classification.which is helpful in preoperative respectability assessment of hilar cholangiocarcinoma.

Objective To investigate the application of OSCE evaluation system on the medical skills examination of clinical medical students and the significance of this system on the training of their medical skills.Methods 20 teachers examed 150 students by the OSCE evaluation system with 4 test stations,by comparing the score of the students of different test stations by one-way ANOVA and evaluating the system by questionnaire survey with Likert 5 on the degree of satisfaction and Likert 3 on effects and intrinsic influencing factors of the system.Results The score of the first and forth test stations was lower than that in the other stations(P＜0.05).8/5.48％ students and 1/5％ teachers were not satisfied with the system.The OSCE evaluation system could exam the psychological diathesis,ability of communication,cooperation,and clinical thinking,practical skill of the students and its effects are moderate (the score was more than 2.0).Evaluation on the intrinsic influencing factors:Students considered the questions were more difficulty in the 2nd,3rd,1st,4th test stations order.4/20％ teachers considered the questions of the second test station was easy.8/40％ teachers considered the duration of the second test station was too long.More than 70％ students and teachers considered the other indexes were rational.Conclusion The OSCE evaluation system can play an effective role in directing the teaching and learning.It can also help to culture the comprehensive capacity of the students.We should gradually improve the design of the system by considering the intrinsic influencing factors.

Objective To investigate the ability of electrocardiogram-gated multislice CT(MSCT)in the diagnosis of myocardial bfidging.Methods Fifty-one patients(82 coronary arteries)with suspected coronary artery disease underwent multi-detector row CT,conventional coronary angiography and intravascular ultrasonography as well.The sensitivity,specificity and accuracy of MSCT for the detection of myocardial bridging were determined.The interobserver agreement was calculated by using Cohen's Kappa test.Results A total of 26 tunneled arteries exclusively located near the middle segment of left anterior descending coronary artery were found by coronary angiography and intravascular uhrasonography.Compared to the invasive methods,MSCT correctly detected 23 of 26 myocardial bridges with a sensitivity of 88%(23/26),specificity of 96%(52/54)and accuracy of 94%(75/80).The Kappa value for overall interobserver variation Was 0.62.Two myocardial bridges diagnosed by MSCT were missed with the invasive method.With the results of invasive and non-invasive methods combined as the standard of reference,the overall sensitivity.specificity,and accuracy of MSCT in detecting myocardial bridging were 89%(25/28),91%(21/23),and 90%(46/51),respectively.Conclusion As a non-invasive imaging modality,MSCT is feasible and reliable in the detection of myocardial bridging.

Objective To construct a prokaryotic expression plasmid for CPSIT_p7 gene from Chlamydophila psittaci ( Cps) 6BC strain and to evaluate immunogenicity of the recombinant protein His-CPSIT_p7 and detect its dynamic expression at mRNA and protein levels in HeLa cells during persistent Cps infection.Methods The fusion protein His-CPSIT_p7 was expressed in E.coli BL21 and purified by Ni-NTA affinity chromatography .BALB/c mice were immunized with the recombinant protein to prepare polyclonal antibody for evaluation of the immunogenicity of His-CPSIT_p7 by ELISA.Penicillin sodium was used to establish a model of Cps persistence infection .RT-PCR and Western blot assay were performed to de-tect the expression of CPSIT_p7 at mRNA and protein levels during Cps persistent infection .Results The fusion protein His-CPSIT_p7 was successfully expressed with the use of constructed recombinant expression plasmid pET30 a-CPSIT_p7 and purified .ELISA result showed that the specific antibody titer against CPSIT_p7 reached 1 ∶1 000 000 on the 40th days after immunization .The expression of CPSIT_p7 at mRNA and protein levels were increased in a time-dependent manner in Cps-infected HeLa cells .The peak of mRNA level was reached at the time point of 36 hours after infection , followed by a time-dependent decrease during Cps acute infection .However , the expression of CPSIT_p7 at mRNA and protein levels were not decreased until 60 hours after infection during Cps persistent infection .Conclusion His-CPSIT_p7 protein was suc-cessfully expressed in the prokaryotic expression system and purified , showing an advantage of good immuno-genicity.Highly expressed CPSIT_p7 at mRNA and protein levels were detected during Cps persistent infection.

Objective To explore the effect of heme oxygenase-1 (HO-1) on expressions of hypoxia inducing factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF)and regeneration of hepatic vascular plexus after orthotopic liver transplantation ischemia-reperfusion injury in rats.Methods Theexperimental study was conducted.According to the random number table,240 SD rats were divided into the 3 groups,80 rats in each group.Empty virus group:rats were transfected with the empty virus.Induced group:rats were transfected with HO-1 overexpression adenovirus.Inhibited group:rats were transfected with HO-1 RNAi adenovirus.Rats were made pairs (1 ∶ 1) and established rat liver transplantation model according to two cuffs method.Rats with less weight and with heavier weight were respectively chosen as donor rats and recipient rats,and then recieved tail intravenous injection of adenovirus at 24 hours before operation.(1) Detection of transfection efficiency of adenovirus before operation:HO-1 expression of liver tissue of rats in each group was detected by Western blot at 12 and 24 hours after injection of adenovirus.(2) Liver function test of recipient rats after liver transplantation:liver functions of recipient rats [alanine transaminase (ALT),aspartate transaminase (AST),alkaline phosphatase (ALP),gamma-glutamyl transferase (GGT)] were detected at l,3,7 and 14 days postoperatively.(3) Pathological histology of liver tissue and injury scores of recipient rats in the 3 groups after liver transplantation:paraffin sections of recipient rats in the 3 groups at postoperative 1 and 14 days were stained by HE staining and observed by light microscope,and were evaluated by Suzuki damage score standard.(4) Relative expressions of HIF-1α,VEGF and HO-1 in liver tissue of recipient rats were detected by Western blot.(5) Von Willebrand factor (vWF) in liver tissue of recipient rats at 14 days postoperatively was detected by immunofluorescence staining and small vessels were counted.Measurement data with normal distribution were represented as x ±s.Comparison between groups was analyzed by the independent-sample t test,comparison among groups was done using one-way ANOVA,and pairwise comparison was analyzed by the LSD test.Results (1) Detection of transfection efficiency of adenovirus before operation:the relative expression of HO-1 of liver tissue of rats at 12 and 24 hours preoperatively after injection of adenovirus was 1.08±0.16 and 1.08±0.26 in the empty virus group,1.18±0.21 and 1.39±0.19 in the induced group,0.87±0.26 and 0.57±0.12 in the inhibited group,respectively,with statistically significant differences in different time points (F =4.232,36.513,P＜ 0.05).(2) Liver function test of recipient rats after liver transplantation:level of ALT at 3 days postoperatively in the empty virus group,induced group and inhibited group was (504±67)U/L,(438±47)U/L and (490±39)U/L,with a statistically significant difference (F=3.517,P＜0.05).Levels of ALT,AST and ALP at 7 days posto-peratively were (443±49) U/L,(430± 34) U/L,(455± 38) U/L in the empty virus group and (382± 49) U/L,(372±50) U/L,(394±25) U/L in the induced group and (493±44) U/L,(455±62) U/L,(470±72) U/L in the inhibited group,respectively,with statistically significant differences (F =10.950,5.667,5.398,P＜0.05).Levels of ALT,AST,ALP and GGT at 14 days postoperatively were (394±46)U/L,(361 ±68)U/L,(417 ±17)U/L,(4.5±1.1)U/L in the empty virus group and (283±47) U/L,(288±60) U/L,(332±46) U/L,(2.5±0.5) U/L in the induced group and (446± 43) U/L,(422± 51) U/L,(423± 63) U/L,(4.3 ± 1.3) U/L in the inhibited group,respectively,with statistically significant differences (F=26.906,9.924,8.013,9.279,P＜ 0.05).(3) Pathological histology of liver tissue and injury scores of recipient rats in the 3 groups after liver transplantation:liver cell swelling,loose cytoplasm and a varying quantity of inflammatory cell infiltration in the portal regions in the liver tissue of 3 groups were observed at 1 day postoperatively.A few inflammatory cell infiltrations in the portal regions,basically normal liver cell arrangement and a slightly swelling of liver cell were found in the empty virus group at 14 days postoperatively.Reduced liver cell swelling and basically normal structure of liver lobule were observed in the induced group.There were small patchy or focal necrosis of liver cell,masses of inflammatory cell infiltration in the portal regions and damage of bile duct in the inhibited group.Suzuki score at 1 day postoperatively in the empty virus group,induced group and inhibited group were respectively 6.7± 1.7,6.1 ± 1.2 and 7.6± 1.3,with no statistically significant difference (F=2.257,P＞0.05).Suzuki score at 14day postoperatively in the empty virus group,induced group and inhibited group were respectively 4.0±0.8,2.9± 0.8 and 5.1± 1.4,with a statistically significant difference (F=9.776,P＜0.05).(4) Western blot results:the relative expressions of HIF-1α and VEGF (43 KD) in liver tissue of recipient rats at 1 day postoperatively were 0.21±0.10,0.30±0.12 in the empty virus group and 0.23±0.09,0.34±0.14 in the induced group and 0.17± 0.06,0.29±0.11 in the inhibited group,respectively,with no statistically significant difference (F =0.902,0.410,P＞0.05).The relative expressions of VEGF (24 KD) and HO-1 in liver tissue of recipient rats at 1 day postoperatively were 1.21 ±0.25,0.55±0.12 in the empty virus group and 2.13±0.40,0.72±0.12 in the induced group and 0.91±0.22,0.26±0.07 in the inhibited group,respectively,with statistically significant differences (F=35.158,39.082,P ＜ 0.05).The relative expressions of HIF-1α,VEGF (43 KD),VEGF (24 KD) and HO-1 in liver tissue of recipient rats at 7 days postoperatively were 0.49±0.22,0.46±0.13,0.98± 0.37,0.98±0.37 in the empty virus group and 0.83±0.26,0.63±0.19,1.60±0.33,1.49±0.46 in the induced group and 0.24±0.09,0.30±0.12,0.64±0.18,0.75±0.26 in the inhibited group,respectively,with statistically significant differences (F=16.853,10.021,20.756,8.156,P＜0.05).(5) Immunofluorescence staining results:number of small vessels at 14 days postoperatively in the empty virus group,induced group and inhibited group was respectively 7.9±2.0,10.6± 1.9 and 7.6 ± 1.9,with a statistically significant difference (F=5.921,P＜0.05).Conclusion HO-1 could promote expressions of HIF-1α and VEGF in liver tissue after liver transplantation ischemia-reperfusion injury and regeneration of intrahepatic vascular plexus,and it also alleviate bile duct ischemia-reperfusion injury after liver transplantation.

AIM: To elucidate the pattern of 5-fluorocytosine(5-FC) induced apoptosis and its role in gene therapy for human pancreatic cancer. METHODS: The human pancreatic cancer SW1990 cells(CEA-producing) were infected with recombinant adenoviruses(Adex1CEA-prCD or Adex1CEA-prZ).Cytosine deaminase(CD) expression was examind by western blot. Apoptosis induced by 5-FC in human pancreatic cancer SW1990 cells genetically modified to express cytosine deaminase was investigated by applying electron microscopy, DNA electrophoresis and flow cytometry analysis techniques. RESULTS: The SW1990 cells infected with Adex1 CEA-prCD were treated with 5-FC at 100 ?mol?L -1 for 48 h, cell apoptosis occurred. Typical apoptosis morphological feature appeared and DNA ladder could be demonstrated on DNA electrophoresis. Apoptosis peak was also showed by flow cytometry. Apoptotic cells accounted for 34.6% of the cell population. Cells in G 1, S and G 2/M phase of cell cycle were 64%, 11% and 7%, respectively. CONCLUSION: The apoptosis induced by 5-FC may be a primary mechanism in CD gene therapy for pancreatic cancer.

0.05).Myocardial perfusion defect scores were decreased significantly from 14.8?3.0 to 10.5?1.8(P

OBJECTIVETo verify whether the extracellular domain of kinase domain region (KDR) has anti-angiogenesis activity in vivo.

METHODScDNA was cloned into adeno-associated virus (AAV) vector pSNAV and transfected to baby hamster kidney (BHK) cells. Recombinant AAV was obtained from the cell culture supernatant after adding helper virus. Recombinant AAV-infected human bladder cancer EJ cell line (EJ cells) were injected subcutaneously into Balb-c nude mice. Tumor specimens were removed from the mice, paraffin-embedded and sliced, then stained by immunohistochemistry. Microvessel density (MVD) was determined under a microscope.

RESULTSThe tumor volume developed by EJ cells transfected with the extracellular domain of KDR was significantly smaller (1.70 +/- 0.18 cm(3)) compared with that in the control (5.62 +/- 0.67 cm(3)) (P < 0.05), although tumor developed to be detectable on almost the same time (14.7 +/- 2.4 days vs 14.1 +/- 3.2 days). Further, MVD in the experimental group was lower than that in the control (41.3 +/- 4.8 vs 6.2 +/- 2.1, P < 0.05).

CONCLUSIONThe extracellular domain of KDR could be expressed in nude mouse bladder cancer tissue and inhibit tumor angiogenesis.

Animals ; Cloning, Molecular ; Cricetinae ; Dependovirus ; genetics ; Endothelial Growth Factors ; metabolism ; Female ; Genetic Therapy ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lymphokines ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; prevention & control ; Urinary Bladder Neoplasms ; blood supply ; therapy ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; Vascular Endothelial Growth Factors