Specific scAbs could be obtained through biopanning from phage antibody libraries by use of antigens as target molecules. scAbs specific to thrombin were separated from mouse antibody library by the panning strategy of alternating liquid-solid phase in this paper. Thrombin was biotinylated by photobiotin at first, then avidin-coated magnetic beads were utilized to isolate specific scAbs. The eluted phages were amplified and subject to the second round panning in microtiter plate to remove the unspecific reombinant phages. 4 specific scAbs were separated from 23 phage clones after four rounds of alternative panning.

Objective To evaluate the effect of hydrogen on the postoperative cognitive function in aged mice.Methods Sixty aged male Kunming mice,aged 12-14 months,weighing 35-45 g,were assigned into 3 groups (n=20 each) using a random number table:sham operation group (group Sham),partial hepatectomy group (group PH) and partial hepatectomy plus hydrogen-enriched saline group (group PH-H).On preoperative day 3,during operation and on postoperative day 3,hydrogen-enriched saline 10 ml/kg was intraperitoneally injected once a day in group PH-H,and the equal volume of normal saline was intraperitoneally injected once a day in S and PH groups.The spatial probe test was performed on postoperative day 7 to evaluate the cognitive function.Ten mice were selected on postoperative days 3 and 7 (after the end of the spatial probe test),and blood samples were collected for determination of serum tumor necrosis factor-alpha (TNF-α),interleukin-lbeta (IL-1β) and high-mobility group box 1 protein (HMGB1) concentrations by enzyme-linked immunosorbent assay.The animals were sacrificed after blood sampling,brains were removed and hippocampi were isolated for detection of the expression of TNF-α,IL-1β and HMGB1 by Western blot.Results Compared with group S,the percentage of time of staying at the target quadrant and the number of crossing the platform within 1 min were significantly decreased,the serum concentrations of TNF-α,IL-1β and HMGB1 were increased,and the expression of hippocampal TNF-α,IL-1β and HMGB1 was up-regulated in group PH (P＜0.05),and no significant change was found in the variables mentioned above in group PH-H (P＞0.05).Compared with group PH,the percentage of time of staying at the target quadrant and the number of crossing the platform within 1 min were significantly increased,the serum concentrations of TNF-α,IL-1β and HMGB1 were decreased,and the expression of hippocampal TNF-α,IL-1β and HMGB1 was down-regulated in group PH-H (P＜0.05).Conclusion Hydrogen can improve the postoperative cognitive function in aged inice.

Objective To investigate the impact of desmoglein 3 (Dsg3) on the proliferation of peripheral T lymphocytes from patients with pemphigus vulgaris (PV).Methods Peripheral blood mononuclear cells (PBMCs) were obtained from 12 patients with PV and 22 normal human controls,cultured with or without the presence of Dsg3 or phytohemagglutinin for 3 days.Flow cytometry was performed to detect the changes of T-lymphocyte subsets in PBMCs stimulated with Dsg3 and proliferation of T-lymphocytes.Results In patients with PV,the percentage of Th2 and Th1 cells was 12.17%±5.32% and 4.08%±1.50%,respectively in Dsg3stimulated PBMCs,9.84%±5.41% and 3.91%±1.38%,respectively in non-stimulated PBMCs.Increased percentage of Th2 cells was observed in Dsg3-stimulated and non-stimulated PBMCs from patients with PV compared with those from normal human controls (both P<0.05).After stimulation with Dsg3,there was a significant proliferation of T cells from patients,and the proliferation rate of CD4+T cells was 4.65%±3.28%,which was significantly higher than that from normal controls(P<0.05).Conclusion Dsg3 can induce the specific proliferation of CD4+T cells,especially Th2 type CD+4 T cells,from patients with PV.

Coronary Artery Disease ; pathology ; Humans ; Medicine, Chinese Traditional

Objective:To investigate the relationship between pemphigus vulgaris(PV) and HLA-DR,DQ haplotypes in Han nations of northeast China.Methods:Polymerase chain reaction-sequence specific primers(PCR-SSP) method was used to detect the HLA-DRB1 and DQB1 alleles of 27 PV patients of Han nation of northeast China, analysed haplotyes and compared with 99 healthy controls.Results:Compared with control group, the frequencies of the haplotypes of HLA-DRB1*140x-DQB1*0503,DRB1*140x-DQB1*0201,DRB1*120x-DQB1*0503 and DRB1*140x-DQB1*0302 increased significantly in PV group. After statistical test, the difference between the two groups was significant.Conclusion:The special haplotypes may contribute to genetic susceptibility to PV in northeast Chinese.

0.05).But GR number[sites/cell]and the expression of GR mRNA in PBMCs from PM/DM was significantly lower than those in healthy controls(P

Objective:Osteoprotegerin (OPG) is a key molecule negatively regulating osteoclast differentiation and activation; and the conserved mycobacterial heat shock 70 (HSP70) peptide p111-125 has also been found to inhibit inflammation reactions in chronic arthritis. BaculoDirectTM baculovirus expression system was selected to express recombinant OPG-HSP70 in insect cells.Methods:The human functional fragment (p22-194) of OPG and functional fragment (p111-125) of mycobacterial HSP70 gene were cloned into the transfer vector pENTRTM/SD/D-TOPO. The recombinant plasmid was performed an LR reaction with the BaculoDirectTM Linear DNA to generate recombinant baculovirus DNA. The cultured Sf9 insect cells were directly transferred with the recombinant baculovirus DNA,and the pure recombinant baculovirus was obtained. Then recombinant baculovirus was infected Sf9 insect cells again to express the OPG-HSP70 gene.Results:The target protein was detected at the time of 48h post infection,reached at highest yield at the time of 72h post-infection. A 28kDa protein immunostaining band was detected by Western blotting from lysate of those cells.And the purified protein was obtained by using Ni-NTA system. Functional stuies on the fusion protein showed it significantly reduce osteoclast cell number[(3.10?0.640) cells under each microscope field in treatment group by comparing to (10.70?0.817)cells in the control group] in the osteoclast inhibition test,and reduce the inflammation reaction in a delayed type hypersensitivity (DTH) mice model (P

Objective To compare the sensitivity of fluorescent quantitation polymerase chain reaction (fluorescent quantitation method) and gene‐chips typing method(gene‐chips method) in the detection of human papillomavirus(HPV) ,and to analyse differ‐ences and clinical significance .Methods A total of 246 women were selected as subjects ,among them ,111 cases of cervical exfolia‐ted cells and 135 cases of cervical tissues were collected and detected .15 kinds of high‐risk HPV genetypes were detected in all sub‐jects by using fluorescent quantitation method and gene‐chips method respectively ,and the detection results were compared . Results The sensitivity of the fluorescent quantitation method in detecting HPV was 55 .28% and that of the gene‐chips method was 55 .69% ,there was no statistically significant difference in sensitivity between the two methods (P>0 .05) .The two methods had relative high conformance(κ=0 .745) .The positive rate of HPV infection was increased with the progression of cervical dis‐ease .Conclusion The fluorescent quantitation method and the gene‐chips method have a relative high conformance ,and both with high sensitivity in detecting HPV .The severity degree of cervical cytological and histological changes may be positively correlated with HPV infection .

BACKGROUND:There are various therapies for children limb fractures involving the epiphysis or the metaphysis. According to the different methods, studies on the growth of the epiphyseal plate are a lot, most of which focus on the effects of Kirschner wires with different diameters or holow screw internal fixation on the development of epiphyseal plate. However, there are rare studies on the influence of cross-epiphyseal plate internal fixation on the growth of epiphyseal plate as wel as the influence level. OBJECTIVE:To prepare a peri-epiphyseal fracture model in young rabbits and to observe the effects of cross-epiphyseal plate implantation and removal on the growth of epiphyseal plate. METHODS: Traverse fracture models were made 5 mm above the right femoral distal epiphyseal plate of 60 young rabbits, and then fixed with suitable “L” steel plate and four screws across the epiphyseal plate and peri-epiphyseal fracture line. The left side served as control. Eight rabbits were kiled and observed at 2, 4, 8, 12 weeks after modeling, respectively, to take out the femoral specimens for measurement of femoral length, thickness of the epiphyseal plate, and number of mastocytes per unit column. Histopathology observation was done and changes in mastocytes and thickness of the epiphyseal plate were detected. Another seven rabbits were selected to remove the metal plate, continued to feed for 2 weeks and finaly executed to observe the above-mentioned indexes. RESULTS AND CONCLUSION: (1) There were significant differences in the above indexes between the plate and control groups at 4, 8, 12 weeks after modeling (P < 0.05 orP< 0.001) but not at 2 weeks after modeling (P> 0.05). These findings indicate that within 2 weeks after cross-epiphyseal plate internal fixation, proper pressue has no remarkable influence on the growth of epiphyseal plate; but after persistent internal fixation (> 4 weeks), the growth of epiphyseal plate can be partialy or completed retarded. (2) At 2 and 4 weeks after modeling, the plate was removed, and 2 weeks later, the femoral length, thickness of the epiphyseal plate and mastocyte counting per unit column were improved to different extents, and there were no differences between the plate and control group (P > 0.05). At 8 and 12 weeks after modeling, the plate was removed, and 2 weeks later, the femoral length and thickness of the epiphyseal plate were shortened, and the number of mastocytes per unit column was decreased obviously, which significantly differed from the control group (P < 0.001). These findings indicate that the chondrocytes in the proliferative and hypertrophy layers lose the differentiation and proliferation abilities, and the femoral length and epiphyseal plate thickness are difficult to recover.

Objective To evaluate the efficacy and safety of linezolid in the treatment of severe lung infections caused by methicillin-resistant Staphylococcus aureus (MRSA).Methods Fifteen patients admitted to ICU due to severe lung infection caused by MRSA received linezolid treatment. WBC, lactic acid (LAC), IL-1β, IL-6, and TNF-α levels were measured before and after treatment. Results Clinical efficacy rate was 73.3%. The level of WBC, LAC and inflammatory cytokines decreased significantly after linezolid treatment (P<0.01).Conclusions Linezolid shows good efficacy in the treatment of severe lung infections caused by MRSA.