Objective To prepare and identify anti‐CD47 monoclonal antibodies .Methods The gene fragment of CD47 was am‐plified by polymerase chain reaction and cloned into prokaryotic expressing vector pET‐32a(+ ) .Purified reconstructed protein was used to immunize BALB/c mice .The immunized spleen cells were isolated and fused with Sp2/0 cells .After screened ,hybridomas secreting anti‐CD47 monoclonal antibody were acquired .Biological activities of antibodies were investigated by Western blot and flow cytometry .Results The recombinant CD47 extracellular domain protein was successfully expressed in BL21 ,and certificated by sodium dodecyl sulfate polyacrylamide gel electropheresis and Western blot .Data of flow cytometry detection demonstrated that the antiserum had high affinity to CD47 protein .Conclusion Recombinant CD47 and its monoclonal antibody ,with high affinity , were successfully prepared ,which could provide reliable tools for the future study of CD47 .

AIM: To probe into the role of 1, 4, 5-trisphosphate inositol (IP_3) and survivin protein in apoptosis of HepG2 cells induced by genistein. METHODS: HepG2 cells were treated with 60 ?mol/L genistein for 12 h, 24 h, 48 h and 72 h. IP_3, survivin and apoptosis rate were assayed by IP_3-[~3H] Birtrak assay, Western blotting and flow cytometry, respectively. RESULTS: IP_3 in groups incubated for 12 h, 24 h, 48 h and 72 h with 60 ?mol/L genistein were significantly lower than that in control (P

AIM:To investigate the role of 1,4,5-trisphosphate inositol(IP3)and Fas gene expression in apoptosis of HepG2 cells induced by quercetin.METHODS:HepG2 cells were treated with quercetin at different concentrations(including 20,40,60,80 ?mol/L)for 72 h and treated with 60 ?mol/L quercetin for 6 h,12 h,24 h,48 h and 72 h.IP3,Fas mRNA,Fas protein and apoptosis rate were assayed by IP3-3H Birtrak assay,RT-PCR,Western blotting and flow cytometry,respectively.RESULTS:When HepG2 cells were incubated with different concentrations of quercetin for 72 h,the IP3 content was lower than those in control.Fas mRNA expression,Fas protein expression and the apoptosis rate were higher than those in control.When HepG2 cells were incubated with quercetin for 6 h,12 h,24 h,48 h,72 h,the IP3 contents were lower than those in control incubated with 60 ?mol/L quercetin for 12 h.Fas mRNA expression was higher than that in control incubated with 60 ?mol/L quercetin for 12 h.Fas protein expression was higher than that in control.The apoptosis rate was significantly higher than that in control incubated with 60 ?mol/L quercetin for 24 h(P

AIM: To probe into the role of 1,4,5-trisphosphate inositol(IP3) and bcl-2 gene expression in inhibiting hepatocellular carcinoma of nude mice by genistein.METHODS: Animals with hepatocellular carcinoma were treated with genistein 1 mg?kg-1?d-1(ip) for 3 weeks.The volume and weight of tumaor were measured.IP3,bcl-2 mRNA,Bcl-2 protein were assayed by IP3- Birtrak assay,RT-PCR,Western blotting,respectively.RESULTS: The tumor volume and weight of animals treated with genistein were lower than those in control(42.7mm3?27.8mm3 vs 52.3mm3?26.5mm3,42.7mg?27.8 mg vs 91.3mg?31.4 mg).IP3 content was lower than that in control [(13.4?1.4)nmol/g protein vs(35.3?6.6)nmol/g protein].bcl-2 mRNA expression was lower in group treated with genistein than that in control(RI which was the gray degree multiply area of bcl-2 / the gray degree multiply area of ?-actin 0.48?0.02 vs 0.56?0.15).Bcl-2 protein expression was lower in group treated with genistein than that in control(RI 1.69?0.52 vs 1.37?0.48).CONCLUSION: Genistein inhibits growth of transplanted hepatocellular carcinoma in nude mouse liver by reducing IP3 production and down-regulating bcl-2 gene expression.

Objective To investigate effect and mechanism of Xuebijing administration at various concentrations on ischemia reperfusion injury (IRI) of rabbit limbs.Methods Thirty New Zealand rabbits were divided into control group (n =10),Xuebijing group Ⅰ (n =10) and Xuebijing group Ⅱ(n =10) according to random number table.Rabbit models of IRI in lower extremities were established.Each group received corresponding therapy after reperfusion:rabbits in Xuebijing group Ⅰ were firstly administered 4 ml/kg Xuebijing solution and 6 ml/kg isotonic saline; rabbits in Xuebijing group Ⅱ were administered 2 ml/kg Xuebijing solution and 8 ml/kg isotonic saline; rabbits in control group were simply administered 10 ml/kg isotonic saline.Venous blood samples were collected before reperfusion and at 1 h,2 h,4 h after reperfusion to measure coagulation parameters (APTT,Fib,INR and PT) and biochemical items (ALB,LDH and CK).Results APTT in Xuebijing group Ⅰ presented obvious improvement at 1 h and 4 h after reperfusion as compared with control group (P ＜ 0.01).PT in Xuebijing groups Ⅰ and lⅡ was significantly longer after reperfusion than that before reperfusion (P ＜0.05).Fib level in Xuebijing group Ⅰ was much higher at 4 h after reperfusion than that before reperfusion (P ＜ 0.05).ALB level at 1 hour after reperfusion showed no statistical differences from that before reperfusion in Xuebijing groups Ⅰ and Ⅱ (P ＞ 0.05).LDH and CK levels in Xuebijing group Ⅰ were much lower than those in control group after reperfusion (P ＜ 0.05).Conclusions Xuebijing injection relieves limb IRI,with better effect in Xuebijing group Ⅰ than in Xuebijing group Ⅱ.Therapeutic mechanism may be associated with its involvement in adjusting clotting function and mitigating injury of muscle tissues.

Paired basic amino acid cleaving enzyme 4 (PACE4),a subtilisin-like endoprotease,which is thought to play a significant role in tumor occurrence and development.Its over or low expression may lead to enhanced proliferation and invasion of tumor cells,and even increase the malignant degree.The specific regulation according to the roles of PACF4 expression in different tumors may be helpful for tumor treatment and prognosis improvement.

Objective To compare the effect between problem-based learning and lecture-based learning for clinical teaching in the department of cardiovascular medicine. Methods Totally 110 five-year-program cardiovascular interns from June 2011 and June 2012 were selected. They were randomly divided into the PBL group(n=55) and LBL group(n=55). PBL and LBL teach-ing methods were applied in the two groups respectively. Teaching effects were evaluated by exam and questionnaire investigation. SPSS 15.0 was used to do data processing; t test was used to compare the average score of two groups;chi-square test was used to process the results of the questionnaire. P<0.05 signifies sta-tistically significant differences. Results There were statistical differences in examinational average score between PBL group and LBL group ((87.89 ±5.39) vs. (82.63 ±5.26), P<0.05). PBL group had significantly higher satisfaction rate in motivating study interests , deepening understanding of theoretical knowledge, cultivating self-learning ability, training verbal expression and developing clin-ical thinking, etc(P<0.05). Conclusions PBL teaching method demonstrates advantages in teaching of cardiovascular medicine and enhances the teaching effect. But the PBL teaching method should be improved in basic knowledge teaching, cultivation of teachers' ability and case selection.

Objective To analyze the virologic responses of peginterferonα‐2a plus ribavirin (Peg IFNα‐2a/RBV) for patients with hepatitis B virus (HBV) and hepatitis C virus (HCV) coinfection by a paired study . Methods Sixteen patients with HBV/HCV coinfection and 32 patients with HCV monoinfection were allocated . The virological response and biochemical response were compared between the two groups after Peg IFNα‐2a/RBV regimen treatment .Chi‐square analysis or Fisher exact analysis was used to compare the responses .Logistic analysis was conducted to evaluate factors associated with sustained virological response (SVR) .Results The median levels of HCV RNA were 6 .54 lg IU/mL in 16 patients with HBV/HCV co‐infection and 6 .98 lg IU/mL in 32 patients of HCV monoinfection ,which was statistically different (Z= 4 .124 , P< 0 .01) .Rapid virological response (RVR) was achieved in 14 cases ,early virologic response (EVR) in 12 cases ,end‐of‐treatment virological response (ETR) in 14 cases and SVR in 9 cases among patients with HBV/HCV co‐infections ,which were comparable to those of patients with HCV monoinfection (RVR:24 ,χ2 =1 .011;EVR:24 ,χ2 =0 ;ETR:23 ,χ2 =1 .474 ;SVR:21 ,χ2 =0 .400 ,all P>0 .05) .The relapse rate in HBV/HCV‐coinfected patients was significantly higher than that in HCV‐monoinfected patients (35 .7% [5/16] vs 8 .7% [2/32];χ2=4 .142 , P= 0 .042) .In HBV/HCV coinfection group ,7 cases were HBV DNA detectable ,7 cases were hepatitis B e antigen (HBeAg ) positive , 13 cases were abnormal alanine aminotransferase (ALT) levels and 12 cases were abnormal aspartate aminotransferase (AST ) levels . After treatment ,only 4 patients were HBV DNA detectable ,5 cases were HBeAg positive ,4 cases were abnormal ALT levels and 5 cases were abnormal AST levels (P= 0 .458 ,0 .716 ,0 .004 and 0 .032 , respectively) .The median level of hepatitis B surface antigen (HBsAg) was 780 IU/mL and decreased to 250 IU/mL after treatment ,while the difference did not reach statistical significance (Z= 1 .826 ,P=0 .068) .A total of 4 patients achieved complete virologic response for HBV ,3 achieved partial virologic response;2 achieved HBeAg seroconversion .Logistic regression analysis showed that EVR was a positive factor for achieving SVR (OR=35 .2 ,95% CI=3 .55-349 .15 , P=0 .002) .Conclusions HCV remains at low replication level in patients with HBV/HCV coinfection .Virological responses can be achieved by Peg IFNα‐2a/RBV treatment .EVR might be a positive factor associated with SVR .

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 study and compare the effects and mechanisms between the Xuebijing injection and Danshen injection on ischemic-reperfusion injury of rabbit limb .Methods 30 New Zealand rabbits were randomly divided into control group ,Xuebi-jing group and Danshen group ,with 10 in each group .The Crinnion model of ischemic-reperfusion injury of rabbit limb was used . Each group was treated correspondingly after the blood flow was restored .Xuebijing group and Danshen group were treated with 4 mL/kg Xuebijing injection 2 mL/kg Danshen injection in saline with 10 mL total volume .Control group was treated with 10 mL sa-line .Before releasing clip and 1 h ,2 h and 4 h after reperfusion ,the blood samples were collected for detecting clotting function (APTT ,FIB ,PT-INR ,PT) ,biochemistry items (ALB ,LDH ,CK) ,MDA level and SOD level .Results Compared with control group ,the APTT of Xuebijing group in 1 h and 2 h and Danshen group in 4 h after reperfusion improved significantly (P<0 .05) . The PT of Xuebijing group after reperfusion extended significantly compared with the PT before reperfusion (P<0 .05) .The FIB level of Xuebijing group and Danshen group in 4 h after reperfusion was much higher than before (P<0 .05) .The LDH and CK level of Xuebijing group after reperfusion was much lower than that of control group (P< 0 .05) .The MDA level of Xuebijing group in 2 h and Danshen group in 1 h ,2 h after reperfusion was much lower than that of control group (P<0 .05) .The SOD activ-ity of Xuebijing group in 1-4 h and Danshen group in 2 h after reperfusion was much higher than that of control group (P<0 .05) .Conclusion Xuebijing injection and Danshen injection have the relieving effect on the limb ischemic-reperfusion injury by ad-justing clotting function and decreasing free oxygen radicals .In terms of relieving the injury of muscle tissue ,the effect of Xuebijing injection might be better than Danshen .