Purpose The research was conducted to evaluate the stability of N-terminal site-specific PEGylated uricase. Methods The enzyme activity is used as an index to evaluate the thermal stability (at 4-80 ℃) , the pH stability, the ability of avoiding the trypsin digestion and half-life in vivo of PEG-uricase, then it is compared with uricase. Results PEGylated uricase is thermally more stable than uricase between 4 ℃ and 60 ℃, but at 70 ℃, the enzyme activity of both PEG-uricase and uricase decreases sharply. In the test of pH stability, the curve of uricase shows an obvious decrease of enzyme activity at 5 .2-6.0 and 9.2-10.0. The behavior of PEG-uricase indicates that the destabilizing was prevented effectively by PEGylation. The test of anti-trypsin digestion suggests that PEG-uricase retains 70% of its enzyme activity,but uricase only 20% ,200 minutes after being reserved in trypsin solution. Stability in vivo indicates that the half-life of PEG-uricase is 1 530 min and uricase, only 45 min. Conclusion PEGylated uricase has improved thermal stability, the pH stability, ability of protecting from trypsin digestion and stability in vivo.

Objective To evaluate the effects of mechanical stretch on pentraxin-3(PTX-3)mRNA and protein expression in human alveolar epithelial cells (A549 cells).Methods The human lung epithelial adenocarcinoma cells A549(A549 cells)were purchased from cell biology laboratory,Tongji Medical College,Huazhong University of Science and Technology.The cultured A549 cells were inoculated on collagen Ⅰ BioFlex plates and divided into 5 groups(n=3 wells each):group Ⅰ normal control;groupⅡsham mechanical stretch;group Ⅲ mechanical stretch;groupⅣsiRNA and groupⅤ siRNA+mechanical stretch.In group Ⅲ the cells underwent square cyclic mechanical stretch for 4 h using the Flexercell Systcm.In group Ⅳ the cells were transfected with chemosynthetic PTX-3 specific siRNA by RNAi technique.In group Ⅴ at 24 h after being transfected with PTX-3 siRNA the cells underwent mechanical stretch for 4 h.In groupⅡ mechanical stretch of the cells were prevented by Flexstep.The expression of PTX-3 mRNA in the cells was detected by real-time PCR and the expression of PTX-3 protein in the culture media was determined by Western blotting.Apoptosis of the cells was measured bv flow cytometry(Beeten-Dickinson,USA).Results PTX-3 mRNA and protein expression was signlficantly up-regulated by mechanical stretch in groupⅢand decreased by transfection with siRNA in group Ⅳand Ⅴ as compared with group Ⅰ andⅡ(P＜0.05 or 0.01).The apoptosis ratio was significantly higher in group Ⅲ and Ⅴ than in groupⅡ and was significantly lower in group Ⅴ thanin group Ⅲ(P＜0.01).Conclusion Mechanical stretch can up-regulate PTX-3 mRNA expression in A549 cells.

Objective To develop an effective and broad immune protective vaccination strategy by using DNA and recombinant vaccinia-based H5N1 vaccines.Methods BALB/c mice were immunized with various prime-boost regimens by using different DNA ( pIRES-based or pVRC-based) and recombinant vaccinia (Tiantan strain, rTTV)-based H5N1 vaccines expressing multivalent antigens (HA, NA, M1 and M2).The differences of immunity induced by two DNA vaccines were compared between intradermal electro -poration (IDE) and intramuscular electroporation (IME) deliveries.Immune responses were analyzed by hemagglutination inhibition( HAI) assay, neuraminidase ( NA)-specific antibody measured by ELISA , mi-croneutralization assay and IFN-γELISPOT assay .Results High levels of humoral immunity and T cell re -sponses were induced in mice primed with DNA-based vaccine than those primed with rTTVb-ased vaccine . DNA priming by IDE resulted in higher levels of neutralizing antibody in mice than those by IME delivery . Higher levels of HAI and anti-NA antibodies as well as NA-specific T cell responses were induced by pVRC-based DNA prime than those by pIRES-based DNA prime .HA-specific T cell responses were also enhanced in mice primed with pVRC-based DNA than those primed with pIRES-based DNA by IME .Conclusion The prime-boost strategies by using DNA-based vaccine in combination with rTTV-based H5N1 vaccine could induce humoral and T cell responses in mice against multi-antigens .Immunities induced by vaccines in com-bination might be modulated by various prime regimes .The study provided references for the further develop-ment of novel H5N1 vaccine and the optimization of immunization programs of combined multi-antigen vac-cine candidates .

Objective To observed the immigration and proliferation of Schwann cells in acellular xe- no-nerve graft in rats.Methods The sciatic nerves on the right side of the rats were exposc.d and 0.8cm long segments of the nerves were removed from the mid-thigh level and replaced by 1.0cm long rabbit nerves made acellular through chemical extraction.After 4 months,the immigration and proliferation of Schwann cells in the graft were revealed by HE staining,S-100 immunohistochemieal staining and transmission electromicro- scope.Results In the rats repaired by acellular nerves,regenerated axons upgrow into the graft,and a- round regenerated axons there were abundant cells aligned,the cytoplasm of which were S-100 immunoreac- tive.Electromicroscope observing showed that regenerated axons were surrounded by myelin formed by the mi- grated cells reoccupied the acellular segments.Conclusion The host Schwann cells can immigrate into rab- bit nerve grafts made acellular through chemical extraction and form myelin enwrapping regenerated axons in rats.

AIMTo probe into the operation mechanism of stress, through the studies on the effects of bile secretion in rats at the condition of water immersion restraint.

METHODSThe animals were divided into six groups (n=8): Group A: restraint alone under room temperature + saline; Group B: water immersion restraint + saline; Group C: restraint alone under room temperature + Atropine; Group D: water immersion restraint + Atropine; Group E: restraint alone under room temperature + Phentolamine; F group: water immersion restraint + Phentolamine.

RESULTSCompared with group A, the capacity of bile secretion in group B decreased significantly (P < 0.05), changes of bile increased remarkably (P < 0.01), but there were no significant decreases on the capacity of bile secretion in group C (P > 0.05) compared with A, Group C only decreased appreciably. Compared with group A, the capacity of bile secretion in group E decreased appreciably (P < 0.05). Compared with group B, the capacity of bile secretion in group D decreased significantly (P < 0.05), pH of bile had no significant changes in group D. Compared with group B, the capacity of bile secretion in group F decreased significantly (P < 0.05), pH of bile had no significant changes in group F. Compared with group D, the capacity of bile secretion and pH of bile in group F had no significant changes.

CONCLUSIONWater immersion restraint stress inhibited evidently on the capacity of bile secretion, and the capacity of bile secretion in water immersion groups decreased significantly, moreover pH of bile increased greatly. At the condition of restraint alone under room temperature, vagus and sympathetic nerve had no significant effects on the bile secretion, but they played important roles in decreases of bile secretion evidently induced by water immersion restraint stress in rats (P < 0.05).

Animals ; Bile ; secretion ; Immersion ; Liver ; secretion ; Male ; Rats ; Rats, Wistar ; Restraint, Physical ; Stress, Physiological

OBJECTIVE:To evaluate the characteristics and regularity of adverse drug reactions (ADR) in children's hospital. METHODS:The clinical drug use and ADR of 3 653 children during 2006～2008 in our hospital were analyzed statistically by designing a child medication registration form. RESULTS:The incidence of 498 ADR cases was about 13.63%; ADR were mostly occurred in 1～3 year-old children(36.14%); 71.29% were caused by intravenously; 66.87% were induced by antibiotics; ADR were more common in spring and winter. CONCLUSION:The drugs have a dual nature. Moreover,children belong to a special group. The measures such as strictly mastering clinical indications,reducing irrational drug use,sufficient therapy and avoiding or reducing the occurrence of ADR can guarantee children grow healthily.

Anastomosis, Surgical ; Colon, Sigmoid/surgery* ; Colorectal Neoplasms/surgery* ; Humans ; Robotics

Adolescent ; Adult ; Aged ; Biopsy ; Delayed Graft Function ; diagnosis ; pathology ; Female ; Graft Rejection ; diagnosis ; pathology ; Humans ; Kidney ; pathology ; physiopathology ; Kidney Function Tests ; Kidney Transplantation ; pathology ; Male ; Middle Aged ; Young Adult

· ATM:To analyze bacterial spectrum and drug sensitivity in conjunctival sac of cataract patients of Kazak.
· METHODS:A total of 538 cases of conjunctival sac secretion in cataract patients of Kazak were collected.The samples were cultured and their sensibilities to antibiotics were tested.
· RESULTS: The bacterial culture was positive in 214 cases.The positive rate was 39.8%. The variety of pathogenic bacteria were mainly made up of gram positive cocci ( 88.3%), and most of them were Staphylococcus epidermidis ( 66.4%), followed by Micrococcus(9.8%).Sex had no effect on conjunctival bacteria rate in the cataract patients of Kazak, while age, place of residence had an effect on camier rate. The camier rate of conjunctival bacteria was significantly higher in people over 60 years old than that in people with age between 40 to 59 years old.And the people from city had a significant lower bacteria positive rate than those from countryside and pastoral. Most of grams were sensitive to Vancomycin, Teicoplanin, Rifampicin, Duly cloth mildew mutual and Amikacin, the tolerance was less than 20%, and they usually had higher tolerance to Penicillin, Erythromycin, Tetracycline and Chloramphenicol (>70%) .
·CONCLUSlON:Gram positivecocci is the most common bacteria in conjunctival sac in cataract patients of Kazak. Staphylococcus epidermidis was most common, followed by Micrococcus.The germ-carrying rate of conjunctival SAC in Kazakh population is associated with the patient’s age and area of residence.The clinical use of antibacterial drugs should be strictly grasp the indications, to reduce the incidence of bacterial resistance.

Objective To develop an effective and broad immune protective H5N1 vaccine.Methods We first developed two recombinant vaccinia ( Tiantan strain) virus ( rTTV ) based H5N1 vaccines, which consisted of bicistron expressing the hemagglutinin(HA) and matrix protein 2(M2), or bicistron expressing the neuraminidase(NA) and matrix protein 1 (M1). The expression of H5N1 protein in rTTVs was confirmed. We immunized the BALB/c mice twice with two kind of dose ( 104 PFU, 107 PFU)using different combination. Subsequently, we assessed the humoral and cellular immune response in vaccinated mice. Results Our data showed that rTTV-based H5N1 vaccine induced rapidly robust HA- and NAspecific antibody level and IFN-γ secreting form cell(SFC) with either single dose of 107 PFU or twice dose of 104 PFU or 107 PFU. We also detected significant neutralizing antibody and matrix-specific immune response. In addition, we found that immunization with two kind of rTTV-based H5N1 vaccines induced much high level of M2-specific antibody than that with single of rTTV-based H5N1 vaccine. Conclusion rTTVbased H5N1 vaccines in this study elicited board array of immunity and our study offers a promising alternative H5N1 vaccine candidates with favorable potential to prevent various H5N1 pandemic.