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

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 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.

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.

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 feasibility of comprehensive assessment of the whole aorta and coronary arteries (CA) simultaneously with high-pitch 128-slice dual-source CT ECG-gated FLASH protocol.Methods A total of 48 consecutive patients with suspected aortic diseases underwent CTA examination of the whole aorta and CA using a ECG-gated FLASH CT protocol (pitch =3.2) without heart rate (HR) control.Aorta,aortic valves and CA were shown with different post-processing modalities.The image quality of the aorta,aortic valves and CA was evaluated and compared according to HR (low HR group ≤ 65 bpm,high HR group ＞ 65 bpm).The scan time,effective dose and contrast medium volume were recorded.Inter-observer differences were calculated by Kappa test.Differences between groups were analyzed by Mann-Whitney test with count data and variance test with measurement data.Results All examinations were completed successfully.The image quality was acceptable in the aorta,aortic valve (100％,48/48) and CA (94.0％,551/586).Percentage of diagnostic quality images was slightly higher in low HR group (94.7％,232/245,93.5％,319/341,Z =-2.504,P ＜ 0.05).Inter-reader reproducibility was 95.8％ (46/48) in the aorta and aortic valve,96.6％ (566/586) in CA,yielded good agreement (Kappa =0.81 and 0.89).The mean attenuation of aorta and CA were higher than 300 HU,especially in low HR group (357.0-446.0) HU.Except proximal segment of right CA,the CNR of whole aorta and coronary arteries were significantly higher in the low HR group (24.5-29.0) than that in the high HR group (20.0-23.1,P ＜0.05).SNR of the whole aorta was higher in the low HR group (13.7-17.9) than the high HR group (11.5-13.9,P ＜ 0.05).The mean scan time was (1.56 ± 0.08) s,effective dose was (4.12 ± 1.23) mSv (2.77-6.77 mSv),and contrast medium volume was (72.8 ± 2.1) ml.Conclusions CT angiography of whole aorta and coronary arteries could be performed simultaueously within 2 seconds with CT ECG-gated FLASH protocol.The images were of diagnostic quality for aortic and CA disease with low dose of radiation and contrast media.However,high HR could decrease the image quality of CA.

Objective To test plasma levels of uPA,uPAR,D-dimer,IL-6 and TNF-α,and observe the relationships between uPA,uPAR and D-dimer,IL-6,TNF-αin patients with SIRS.Methods A prospective,clinical case-control study was adopted.Cases were collected from our hospital in January 2008 to January 2010,and all were ＞ 55 years of age.Venous blood samples were collected via routine venipuncture.Eighty-five patients were divided into two groups according to diagnostic criteria of SIRS:SIRS group collected from Intensive Care Unit ( n =50) and non SIRS group collected from medical ward ( n =35).The control group comprised 30 unrelated healthy blood donors who visited the General Health Checkup Division at our hospital.Patients with (1) pregnant women; (2) cancer; (3) died after admitted into ICU in 7 days; (4) after cardiopulmonary resuscitation ; ( 5 ) with previous blood system diseases; (6) patients with SIRS before admitted into ICU were excluded from the study.uPA,uPAR,D-D,IL-6 and TNF-α in blood were detected by commercial enzyme-linked immunosorbent assay (ELISA) kits.The data was analyzed using SPSS version17.0.Data accorded with normal distribution of measurement was expressed as mean ± standard,and analyzed by independent-samples t test; non-normal distribution of measurement data,expressed by median,was analyzed with Mann-Whitney test.Relationships between plasma levels of uPA,uPAR and D-dimer,IL-6 TNF-α were analyzed using Spearman rank correlation test.To compare with blood level of uPA,uPAR,IL-6 and TNF-α in SIRS patients in the application of diagnostic value of MODS,we constructed the receiver operating characteristic curve ( ROC curve) for blood levels of uPA,uPAR,IL-6 and TNF-α in 24 h.Results The plasma levels of uPA,uPAR,D-dimer,IL-6 and TNF-αin patients of SIRS were obviously higher compared with non-SIRS and normal controls ( all P ＜ 0.01 ).Correlation analysis showed that there was positive correlation between uPAR level and IL-6 level (r =0.395,P =0.004) ; there was positive correlation between uPAR level and TNF-αlevel ( r =0.606,P ＜0.01 ).There was no correlation between uPAR levd and D-dimer level ( r =- 0.069,P =0.632).There was no correlation between uPA level and D-dimer,IL-6 or TNF-α ( all p ＞ 0.05).There ROC curve were made based on the abilities of uPAR,D-dimer,IL-6 and TNF-αlevels in 24 hours to diagnose MODS,and the ROC areas under the curves were 0.76,0.58,0.86,0.83 respectively.Conclusions These results demonstrate that uPA and uPAR play a major contributory role in patients with SIRS in the process of coagulation disorders,but the mechanism in SIRS is not the same.uPAR may play a central rolein the development of SIRS to MODS.

Objective To investigate whether IL-17 could stimulate the vascular endothelial growth factor (VEGF) production on HaCaT cells alone. We also investigated whether shikonin could inhibited the proinflamation effects of interleukin-17(IL-17) acting on HaCaT cells. MethodsWe examined the expression of VEGF by double antibody sandwich enzyme-linked immunosorbent assay ( ELISA ) and realtime polymerase chain reaction(RT-PCR) in HaCaT cells and the cell supernatant. The viability of HaCaT cells in the drug group was detected by the Cell Counting Kit-8 (CCK-8). ResultsThe expression of VEGF in different time IL-17-stimulated groups on HaCaT cells and the cell supernatant were higher than the control group( P＜0.001 ). The expression of VEGF in different drug treatment groups on HaCaT cells and the cell supematant were lower than the stimulated group by IL-17 ( P＜0. 001 ). The cell viability of different drug treatment groups have no significant difference( P＞0.05 ). ConclusionWe show that IL-17 specifically and time-dependently augmented and induced VEGF expression on HaCaT cells and the cell supernatantThen shikonin markedly inhibited the increase tengency of IL-17 effection on HaCaT cells and the cell supematant level.

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

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