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.

Selective salpingography and transvaginal catheter dilatation using a self- made coaxial catheter set were performed in 45 women with infertility and fallopian tube ob- struction.The successful catheterization rate was 95.5% for fallopian tubes and the recanal- ization rate proximal allogian tubes was 80%,Normal intrauterine pregnancy was achieved in total 15 women(33.3%)with in two years follow-up 50% and 17.4% pregnant rates were obtained in secondary infertility and primary infertility,respectively.The authors be- lieve that transvaginal catheter dilization is a useful method for female infertility with fallopi- an tube obstruction,especially for the secondary infertility and proximal fallopian tube ob- struction.The self-made coaxial catheter set holds the advantages of effectiveness,safety, and inexpensiveness.

Objective To compare the biomechanical performance of three different fixation instruments for tibial fractures.Methods Fourteen fresh tibial specimens were made into models of oblique fracture.Seven models were fixed with unilateral axial dynamic fixation(UADF),and seven with limited contact dynamic compression plate(LC-DCP).After biomechanical tests had been done for the UADF group,an extra screw was used at the oblique fracture site to reinforce the fixation with extra limited internal fixation.Each model was tested for its hiomechanical performance in resisting compression,bending and rotation.Results The performance of UADF was significantly poorer than that of LC-DCP and UADF with limited internal fixation in anti-compression,an- ti-bending and anti-rotation(P＜0.05).There was no significant difference in the rigidity between LC-DCP and UADF with limited internal fixation(P＞0.05).Conclusion UADF with extra limited internal fixation is a recommendable method for tihial fractures because it cart provide the same effective fixation as LC-DCP does.

Objective To establish a method for the preparation of long-circulating liposomes (LCL) containing etoposide and to observe its stability in rats plasma.Methods The etoposide-containing liposome was prepared by ethanol injection method. Polyethylene glycol-distearoyl phosphatidylethanolamine (PEG2000-DSPE) was used to modify the membrane of the liposome. Reversed-phase high pressure liquid chromatography (RP-HPLC) was used to detect the concentration of the liposome,and dynamic release method was used to study its stability in mice plasma.Results The mean size of the LCL containing etoposide was (180?26) nm,and the mean concentration of etoposide was (4.78?0.22) mg/mL,with the entrapment efficiency being (88.71?8.2)%. The leakage ratio of the conventional liposome containing etoposide and LCL containing etoposide in mice plasma were (80.14?1.59)% and (46.72?2.61)%,respectively.Conclusion LCL containing etoposide with high entrapment efficiency and low leakage rate can be obtained by using ethanol injection method. Additionally,modification by PEG2000-DSPE could raise the stability of the liposome.

Objective To study the damaging effect of tumor necrosis factor-alpha (TNF-?) and Interleukin-1?(IL-1?) on brain of rats in acute pancreatitis(AP). Mothods AP pancreatitis model in rats was induced by retrograde injection of 5% sodium taurocholate into the pancreatobiliary duct(1ml/kg body weight), then pretreated with normal saline (NS) or TNF-??IL-1?, TNF-? plus IL-1? respectively. Animals were killed at 5 h after drugs administration. The changes of brain water contents, leukocyte accumulation and adhesion were measured, and pathological studies of pancreas and brain were also performed. Results In groups with inflammatory cytokines pretreated, brain water contents, leukocyte accumulation and adhesion increased more significantly than those in control group and in NS pretreated group(P0.05). Conclusions Inflammatory cytokines such as TNF-? and IL-1? play important roles in the formation and acceleration of the brain damage in acute pancreatitis.

Objective To investigate the expression of vascular endothelial growth factor/vcascular endothelial growth factor receptor(VEGF/VEGFR) in bone marrow of children with acute leukemia(AL),and explore their relationship of the clinical features,and observe changes before and after chemotheropy.Methods The bone marrows of 53 children with AL were assayed to study the expression of VEGF/VEGFR(KDR,Flt-1)before and after chemtheropy with S-P immhistochemical staining.Their relations to the clinical features were evaluated.Results The expressions of VEGF,Flt-1,KDR were significantly higher in newly diagnosed children with AL than those of control group.They were significantly higher in children with acute myeloid leukemia(AML) than in children with ALL.The expressions of VEGF,Flt-1,KDR in remission chidren after chemotheropy were significantly lower than before chemotheropy.There was a positive correlation of the percentage of bone marrow blasts with VEGF expression in children with AL.There was also a positive correlation of the percentage of leukemic cell in blood rountine with VEGF expression.For the untreated group of children,no correlation was found between expressions of VEGF,Flt-1,KDR and age,sex,extramedullary infiltration.Conclusions Expression levels of VEGF,Flt-1,KDR in bone marrow of children with AL increases.VEGF/VEGFR may play an important role in process of chidhood AL.

ObjectiveTo investigate the effect of measures on controlling of schistosomiasis in Jurong city Jiangsu province and provide a scientific basis for doing a better job in prevention and control of schistosomiasis.MethodsMonitoring data from 2005 to 2010 was analyzed,including the effect of spraying drugs,changing environment,coupled with health education measures in eradicating oncomelania.Oncomelania areas,positive detection rate of schistosomiasis,and the awareness rates of prevention knowledge of schistosome were observed longitudinally.Correlation of coverage rate of snail controlling and resident positive rate of schistosomiasis was studied.ResultsThere were no infected snails from 2005 to 2010.The densities of living snails and areas with snails decreased year by year.Residents' positive detection rate of schistosomiasis decreased from 0.34％(36/10 551 ) to 0.00％(0/12 372) from 2005 to 2010.The rates of students,and citizens' knowledge of schistosome on prevention of the disease increased from 88.28％(128/145) and 86.44％(204/236) to 96.58％(141/146) and 92.83％(246/265) from 2005 to 2010,respectively.There was a negative correlation between oncomelania areas and drug coverage areas(r =- 0.910,P ＜ 0.05 ); there was no correlation between oncomelania areas and coverage rate of snail control by environmental modification (r =- 0.404,P ＞ 0.05 ); there was no correlation between positive rate of schistosomiasis in resident and coverage rate of snail control with molluscisides,and coverage rate of snail control by environmental modification (r =0.107,- 0.496,all P ＞ 0.05).ConclusionsSnails areas in Jurong city is decreased year by year.Residents' positive detection rate of schistosomiasis infection is maintained at a low level.The effects of oncomelania-controlling,health education,and chemotherapy are remarkable.

OBJECTIVETo observe the clinical efficacy of Shugan Yiyang Capsules in the treatment of asthenospermia and its action mechanisms.

METHODSWe randomly assigned 135 asthenospermia patients to groups A (n = 47), B (n = 45), and C (n = 43) to be treated with Shugan Yiyang Capsules, oral levocarnitine, or combination of the two. We observed sperm quality and the level of α-glucosidase in the seminal plasma before and after medication.

RESULTSThe total effectiveness rate was 70.21% in group A (markedly effective in 16 cases and effective in 17), 68.89% in group B (markedly effective in 15 cases and effective in 16), and 83.72% in group C (markedly effective in 16 cases and effective in 20), significantly higher in C than in A and B (P < 0.05). Both sperm quality and the level of α-glucosidase in the seminal plasma were improved in the three groups of patients, most obviously in group C.

CONCLUSIONShugan Yiyang Capsules can be used for the treatment of asthenospermia, and its effect can be enhanced in combination with oral levocarnitine.

Asthenozoospermia ; drug therapy ; enzymology ; Biomedical Research ; Capsules ; Carnitine ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Semen ; enzymology ; Spermatozoa ; alpha-Glucosidases ; analysis

Objective To investigate if Rac1 GTPase activation plays an important role in hyperpermeability and tyresine phosphorylation of tight junction induced by vascular endothelial growth factor (VEGF) in glomertdar endothelial cells (GEnCs). Methods Primary cultured rat endothelial cells were used as experimental model. The effect of VEGF at different concentrations (5 or 50 μg/L) on endothelial permeability was investigated by transendothelial electrical resistance (TEER). The permeability of GEnCs transfected with wild type Rac1 (wtRacl) or dominant negative Racl (N17Rac1) was also detected. Immune precipitation and immune blotting were used to detect the tyrosine phosphor-occludin in GEnCs. Results VEGF at high concentration (50 μg/L) induced hyperpermeability in GEnCs (P<0.05). At the same time, GTP-binding and membrane-bound Racl GTPase significantly increased(P<0.01)in GEnCs. Tyrosine phosphor-occludin was also increased (P<0.05) under VEGF stimulation. However, transfection of GEnCs with N17Rac1 dramatically attenuated the effect of VEGF on tyrosine phospho-occludin and endothelial cell permeability. Conclusions Increased VEGF can induce hyperpermeability in glomerular endothelial cells, which is related to occludin tyrosine phosphorylation through Racl activation. It provides a framework for understanding the role of VEGF-induced Racl-phospho-occludin pathway in the integrity of barrier function in the diabetic milieu.