Objective To determine the free thiols in the chimeric protein PfCP-2. 9 of Plasmodium falciparum expressed by Pichia pastoris. Methods Two experiments of reverse phase HPLC and Ellman' s reaction were applied to the PfCP-2.9 for the determination of its free thiols. For RP-HPLC analysis, three kinds of samples were tested: PfCP-2. 9, dithiothreitol-reduced PfCP-2.9 and indoacetic acid-alkylated PfCP-2.9. Results Both experiments showed that there were no any free thiols present in the PfCP-2. 9. Conclution The disulfide bonds between cysteine residues of PfCP-2. 9 were formed completely.

Objective:To express MSP1-31 gene of Plasmodium falciparum in Salmonella typhi CVD908 vaccine strain using a tetracycline-controlled P LtetO promoter. Methods:The MSP1-31 gene was cloned into the plasmid of pZE11 and transformed into the CVD908/tetR strain by electroporation. Expression of MSP1-31 in CVD908/tetR strain was detected using the method of Western blot. Results: The recombinant plasmid of pZE11/MSP1-31 was constructed, there was effective expression of MSP1-31 protein in CVD908/tetR strain in presence of tetracycline, and no expression of gene in absence of tetracycline. Conclusion: The recombinant Salmonella typhi strain in which the expression of Plasmodium falciparum MSP1-31 fragment induced by tetracycline is established successfully.

Objective To study the changed status of mortality from urinary and male genital diseases in Shanghai Hongkou district in recent years. Methods Mortality from diseases in Shanghai Hongkou district population from 1991 to 1998 was studied and analyzed. Results There were 1130 persons died of urinary and male genital diseases,749 being male and 381 female.It accounted for 2.08% of the total death,being 2.58% in the male and 1.51% in the female.The 5 major urinary and male genital diseases were in turn nephritis and nephropathy,bladder cancer,benign prostatic diseases,prostate cancer and renal carcinoma.Mortality from bladder cancer,renal cancer,prostate cancer and other prostatic diseases has been rising as compared to those in the 80s especially protatic cancer and other prostatic diseases whereas mortality from nephritis and nephropathy,pyelonephritis and renal failure was decreasing. Conclusions Mortality from prostate cancer and other prostatic diseases in population is remarkably rising.Attention is called to the study,prophylaxis and treatment of prostatic diseases.

Objective To evaluate the efficacy of oral etopside (VP16) and oral cyclophosphamide (CPM) combination in the treatment of hormone-refractory prostate cancer (HRPC). Methods Between June 2000 and July 2003,9 patients with HRPC were treated with oral etopside (50 mg/d) and oral cyclophosphamide (100 mg/d) for 21 days with every 28 days as a cycle.Inclusion criteria were previous complete androgen blockade,anti-androgen (flutamide) withdrawal evaluation,and clinical or biochemical disease progression.The therapy was continued until there was evidence of disease progression or the patients could not tolerate the adverse effects of the medications. Results All the 9 patients had a mean follow-up of 7.5 months.PSA levels decreased by at least 50%,from pre-treatment of (90.5?43.6)ng/ml to post-treatment of (24.8?22.2)ng/ml,in 4 patients. The mean duration of response was 6.8 months (range,2-15 months).An objective response was obtained in 2 patients (1 of CR and 1 of PR).Toxic and adverse effects were minimal. Conclusions The combination of oral VP16 and CPM may be an efficacious and well-tolerated regimen in patients with HRPC.

50% for more than 1 month as effective,and the efficacy for soft tissue metastases were classified as complete,partial remission,stabilization and progression.Results All patients were followed up for 6-24 months(mean,12 months) with the evaluation of efficacy and toxicity.PSA levels decreased by at least 50% in 6 of 12 cases(50%);it decreased from(63.9?47.3)ng/ml before treatment to(14.4?8.8)ng/ml after treatment,with a mean duration of response being 7.5 months(range,5-12 months).Partial remission of soft tissue metastases was obtained in 2 cases;the metastatic lesions were reduced from 4.0 cm?5.0 cm,(3.0cm?)(3.5) cm to 2.0 cm?2.0 cm,1.0 cm?1.5 cm,respectively,by the treatment,with response duration being 3 and 8 months,respectively.Toxicities were minimal with leukopenia at grade Ⅰ in 1 case,anemia at grade Ⅰ in 1,baldness at grade Ⅰ in 1,nausea at grade Ⅰ in 2 and impaired liver function at grade Ⅱ in 1.Conclusions The combination of oral estramustine phosphate and oral etoposide may be an effective and well-tolerated regimen in patients with HRPC.

BACKGROUND:Alpha-calcium sulfate hemiydrate/biphasic bioceramics (α-CSH/BCP) as a bone repair material has been widely used in the clinic, but there are stil some deficiencies. OBJECTIVE:To observe the effect of α-CSH/BCP on the repair of bone defects of the radius in rabbits. METHODS:The unilateral radius of 18 adult New Zealand white rabbits was selected to establish bone defect models, and then these rabbit models were randomized into three groups that were treated with α-CSH,α-CSH/BCP, BCP materials, respectively. At 4, 8, 12 weeks after implantation, bone samples were taken for gross and X-ray observations. RESULTS AND CONCLUSION: (1) Gross observation: At 12 weeks of implantation, the bone defect was completely repaired in the α-CSH/BCP group, and the defected radius recovered basicaly; in the α-CSH group, bone defects were visible; and in the BCP group, there was stil some material for filing and coverage. (2) X-ray observation: In the α-CSH group, the degradation of implanted materials was faster than new bone formation, and bone defects were visible; in the α-CSH/BCP group, bone defects were healed, cortical bone remodeling was good, and the bone marrow cavity was re-canalized; in the BCP group, the implanted materials were stil visible, and bone defects were not repaired. These findings indicate that the α-CSH/BCP bone repair material can significantly promote the repair of bone defects of rabbit radius, with a better repair effect than α-CSH or BCP alone.

Objective:To evaluate the clinical efficacy and safety ofHJ-RY-1 therapeutic pad ofsoft stone in treating osteoarthritis.Methods:Patients with osteoarthritis with syndrome ofqi stagnancy and blood stasis were enrolled according to the diagnostic criteria and the evaluation standard ofcurative effect in Instruction Principle ofClinical Research ofNew Chinese Medicines, and were randomly divided into experimental group(treating by HJ-RY-1 therapeutic pad ofsoft stone) and the control group(treating by the paster offar infrared bioactive ceramics).Put them respectively on the afflicted part for more than 12 hours a day, and the treatment course ofboth groups lasted 14 days.The main turnover was determined by the improvement of6 clinical parameter indices and the incidence rate ofwhole curative effect and side effect after a treatment of7 days and 14 days.Results:72 patients were randomly equally divided into two groups, and received predetermined interveneing therapy.There were similar baseline characteristics and comparabilities between them.Totally 68 patients finished 2 weeks follow-up observation and 4 cases were missed(33/36 in experimental group;35/36 in the control group).1 weeks later, there were no significant difference in clinical situation and overall curative effect between two groups.But after two weeks interveneing therapy, there were significant difference on activity pain(t-value was 3.3104), the joint movement range(t-value was 3.1596), self-sensation(t-value was 2.5521) and overall curative effect(t-value was 3.5694) between them(P

The importance of cytotoxic T-lymphocyte(CTL)against malaria parasite in pre-erythrocytic stage has been presented in relevant researches. In order to investigate whether one CTL epitope(YLNKIQNSL)involved in a chimeric pre-erythrocytic stage vaccine candidate of Plasmodium falciparum which was expressed and purified in the laboratory can stimulate in vivo CTL response,HLA-A*0201 transgenic mice were immunized with this vaccine candidate. Enzyme-linked immunosorbent spot(ELISPOT)assay was performed on the splenocytes from the immunized transgenic mice. Positive result indicated that this CTL epitope can be in vivo processed and correctly presented.

Objective To prepare and characterize monoclonal antibody against a malaria vaccine candidate, PfCP-2.9 chimeric protein of Plasmodium falciparum. Methods BALB/c mice were immunized with PfCP-2.9,and the spleen cells were used for fusion with SP2/0 cells. The monoclonal antibodies were analyzed by ELISA,Western blotting as well as growth inhibition assay. Result A monoclonal antibody was obtained. It interacted with the PfCP-2.9 recombinant protein by ELISA and Western blotting. The interaction of the monoclonal antibody with the protein was reduction-sensitive,indicating that the antibody recognized a conformational epitope. Moreover,the antibody also recognized the cultured parasites of P.falciparum by indirect immunofluorescent antibody test(IFA). When tested by growth inhibition assay,the antibody significantly inhibited parasite growth in vitro of 56% inhibition rate at the antibody concentration of 0.3 mg/ml. Conclusion A monoclonal antibody against PfCP-2.9 malaria vaccine candidate has been obtained,which recognizes a conformational epitope of the protein and natural protein.

Objective To construct a chimeric protein of Plasmodium falciparum pre erythrocyte stage (named as PfCP 4). Methods Thrombospondin related anonymous protein (TRAP) and circumsporozoite protein (CSP) of Plasmodium falciparum have been considered important candidates for pre erythrocytic malaria vaccine. The sequences of ectodomain of TRAP (aa: 26-330) and (NANP) 19 repeat region and entire carboxy terminus of CSP were fused to generate the PfCP 4 via a hinge consisting of Gly Pro Gly. The 1577 bp sequence of PfCP 4 was synthesized by asymmetric PCR based method and the synthetic gene was inserted into pQE. The resulting plasmid was transformed into E. coli SG13009 for inducible expression with IPTG. The expression product was detected by Western blotting. Results The result of Western blotting showed that the entire PfCP 4 recombinant protein was produced under IPTG induction whereas no product was detected in the cell without induction. The molecule weight of the protein was 57 kDa which was identical to the expected size, and the product was recognized by polyclonal antibodies against CSP protein. Conclusion A chimeric protein of Plasmodium falciparum pre erythrocyte stage (named as PfCP 4) was constructed successfully.