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 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 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

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.