To screen and characterize human phage antibody to hepatitis C virus E2 antigen. The recombinant phages were panned by HCV E2 antigen which was coated in a microwell plate. After three rounds of biopanning, 56 clones specific to HCV E2 antigen were obtained. The specificity of scFv was determined by ELISA method and cross reaction with BSA and competitive inhibition assay. HCV E2 phage antibody had a specific combination character with recombinant hepatitis C virus E2 antigen. The DNA sequence data showed that the scFv coding gene was 771bp in size

To construct expressive vector for human ScFv against core protein of hepatitis C virus (HCV core ScFv),and to express soluble HCV core ScFv in E.coli JM109. Using phage display technique, the recombinant phages were panned by recombinant core antigen which was coated in a microtiter plate, and after five rounds of biopanning, 86 clones were identified specific to core antigen. 750 bp fragment could be released from the plasmid of positive phage clones, and the sequence analysis indicated that we have obrained the ScFv DNA fragment. Then DNA fragment was inserted into the expressive vector pCANTAB5E, and E. coli host JM109 was transformed and induced by IPTG. The specificity of ScFv in the culture medium was evaluated by enzyme linked immunosorbent assay (ELISA).The molecular weight of expressed HCV core ScFv protein is 28 000 dalton as determined with the aid of SDS polyacrymide gel electrophoresis (PAGE). The expressed HCV core ScFv protein will be useful in the immunohistochemical study of liver tissue from patients with hepatitis C and gene therapy against HCV infection.

Angina Pectoris ; drug therapy ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Phytotherapy ; Technology, Pharmaceutical

Objective:To study the safety and effect of enteral nutrition in preoperative bowel preparation and postoperative early enteral nutritional support for patients with colorectal cancer.Methods:42 patients with colorectal cancer were randomized into 2 groups.21 patients in the experimental group(enteral nutrition group) were applied with Nutrison Fibre as a kind of liquid diet in preoperative bowel preparation and postoperative early(in 12~24 hours) enteral nutritional support.21 patients in the control group(traditional therapy group) made the bowel preparation by the traditional method of semiliquid-liquid-fasting with infusion and managed with ordinary process after operation.The times of intestinal lavage the evening before operation,the satisfaction of colon cleaning,recovery time of passing flatus,occurrence of ill reaction and complication were observed.Parameters including body weight,hemoglobin(Hb),lymphocyte count(LY),plasma total protein(TP),serum albumin(ALB),prealbumin(PA) were measured and compared before bowel preparation(3th day before operation),before operation and on the 8th day after operation.Results:There were no statistical difference in the two groups about the satisfaction of colon cleaning.In the experimental group,the time of intestinal lavage was fewer(P(0.05).) But in the control group,Hb,LY,TP,ALB and PA were all significantly lower(P

ObjectiveTo observe the effects of acupuncture combined with massage on shoulder subluxation and motor function recovery of the upper extremities in patients with hemiplegia after stroke.Methods60 hemiplegic patients were randomly divided into two groups for treatment with acupuncture cooperates with massage or usual rehabilition, respectively. The recovery of the patient's shoulder subluxation and movement function of upper extremities were evaluated 2 months after treatment. ResultsThe shoulder subluxation and movement function of the upper extremities were improved after treatment with both therapies (P<0.01), and acupuncture cooperates with massage showed better effect (P<0.01). ConclusionAcupuncture cooperates with massage can facilitate the recovery of shoulder subluxation and motor function of the upper extremities after hemiplegia.

Objective To improve the level of diagnosis and treatment of the sharp injury in the back. Methods 47 cases which were treated from Jan 1991 to May 2000 were reviewed. ResultsAmong 37 cases who underwent the exploration, 5 cases died. Among 10 cases who underwent conservative treatment, 2 cases died. ConclusionThe condition of sharp injury in the back is very complicated,it is easy to be misdiagnosed, the mortality is high. Saving should be equalled with the diagnosis and treatment. The application of wound exploration, abdominal puncture, ultrasound examination and X-ray checking is valuable to the diagnosis. The patients with operation indications should be operated at once,while the others should be observed for some time to prevent the delayed clinical manifestation.

Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E　coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.

Objective To investigate the effect of HSP70/CD80 DNA vaccine on the airway inflammation and hyperresponsiveness of asthmatic mice. Methods Forty female healthy BALB/c mice were randomly divided into 4 groups; saline group, asthma group, pcDNA3. 1 plasmid control group, and prevention group with HSP70/CD80 DNA vaccine, with 10 mice in each group. The mice were immunized by intramuscular( i. m. ) injection with HSP70/CD80 DNA vaccine before sensitization and challenge with ovalbumin. Then, the murine model of allergic asthma was made with injection of ovalbumin intraperitoneal ( i. p. ) , and inhalation of ovalbumin. Before mice were sanctified, their airway hyperresponsiveness( AHR) was measured. After mice were sanctified, bronchoalveolar lavage fluid( BALF) was obtained and cytokine IL-13 and IFN-γ were measured. And the lung histology and histochemistry were examined. Results Compared with mice in asthma and pcDNA3. 1 group, mice in vaccine group showed significantly reduced airway inflammation (P＜0. 05) and AHR (P＜0. 05). IFN-γ content in BALF were increased in mice from vaccine group compared with the asthma group and the pcDNA3. 1 group ( P ＜0. 05) , and IL-13 content in BALF were decreased. Conclusion HSF70/CD80 DNA vaccine can reduce airway inflammation and hyperresponsiveness in asthmatic mouse and this chimerical plasmid could be a candidate vaccine to prevent asthma.

Objective:To investigate the expression of HIF-1?and its relationship with angiogenesis in osteosarcoma.Methods: Osteosarcoma MG-63 cells were cultured in vitro under hypoxia and mimic hypoxia conditions.Thirty paraffin-embedded osteosarcoma tissues and 20 fresh frozen osteosarcoma specimens were collected.The mRNA and protein expression of HIF-1?and VEGF were detected by RT-PCR,Western blotting,ELISA,and immunohistochemistry methods.The mean vessel density(MVD)were also calculated.Results:The mRNA level of HIF-1?had no change under hypoxia and minic hypoxia conditions,whereas the protein expression was increased dramaticaly.The mRNA and protein expression of VEGF was significantly increased under hypoxia and minic hypoxia conditions.The positive rate of HIF-1?mRNA(90%)and VEGF(100%)in 20 fresh frozen tissues were higher than those of the para-tumor tissues(P