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 evaluated the improving effect of training method of point-line-plane on development quotient in premature infants with brain injury.Methods 174 cases of premature children with brain injury were randomly divided into study group (89 cases) and control group (85 cases).Control group given Bobath and Vojta traditional rehabilitation training,and the study group was given training method of point-line-plane.Patients in both groups were evaluated synthetically based on The Evaluation Chart on Neu-robehavioral Development of Children Aged 0-6 Years,developed by Capital Institute of Pediatrics,prior to the treatment,and after the first,the second,and the third course of the training programs,respectively.Re-sults The score of developmental quotient ( DQ) between study group and control group was no significantly different(P>0.05).Compared with before treatment, the scores of DQ was significantly increased after the first, the second and the third course of training treatment in study group and control group((82.71±12.28) vs (81.17±9.63), t=0.859, P>0.05;(89.65±10.02) vs (87.02±7.39), t=1.747, P>0.05). Compared with the control group, the scores of DQ in study group was no significant difference after the first and the second course of treatment,but there was significantiy different after the third course treatment((95.26±8.87) vs (91.31±7.63), t=2.945, P=0.004).Conclusion The training method of point-line-plane can improve the clinical efficacy of premature infants with brain injury at the early phase, which is worth widely applying in clinical practice.

Objective To explore the role of 1, 4, 5-trisphosphate inositol ( IP3) and bax gene expression in apoptosis of HepG2 cells induced by Quercetin.Methods HepG2 cells were cultured for 72h as control. HepG2 cells were treated with different concentrations including 20,40.60,80?mol/L Quercetin for 72h, and treated with 60?mol/ L Quercetin for 12h, 24h, 48h and 72h. IP3, bax mRNA, bax protein and apoptosis rate were assayed by IP3-[3H] Birtrak assay,RT-PCR, Western blotting and flow cytometry, respectively.Results In HepG2 cells incubated with each of the concentrations of Quercetin for 72 h,IP3continent was lower than that of control(P

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 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 .

Hepatocellular carcinoma is one of the largest causes of cancer-related deaths worldwide for which there are very limited effective treatment options.The ubiquitin-proteasome pathway(UPP) has rapidly become acknowledged as both critical mechanism for cellular biological function and a latent target of regulation of cancer-related disease.This review focus on the role of ubiquitin-proteasome pathway in hepatocellular carcinoma and its correlation factors(HBV、P27、NF-?B,et al),in order to find novel therapeutic interventions against the genesis and development of hepatocellular carcinoma.

Objective To elucidate the constituents from the root of Euphorbia pekinensis and to look for new products with antitumor activity.Methods The chromatography on silica gel was used and the structures were identified by IR,NMR,and MS spectral analyses and their physicochemical properties were comparied.Results Eight compounds were isolated and identified as octadecanol(Ⅰ),octadecanyl-3-methoxy-4-hydroxybenzeneacrylate(Ⅱ),?-sitosterol(Ⅲ),triacontanoic acid(Ⅳ),2,2′-dimethoxy-3,3′-dihydroxy-5,5′-oxo-6,6′-biphenylformic anhydride(Ⅴ),euphol(Ⅵ),tirucallol(Ⅶ),and euphpekinensin(Ⅷ).Conclusion The compounds Ⅰ,Ⅳ,Ⅵ,and Ⅶ are isolated from the roots of E.pekinensis for the first time.