Objective To study the biological characteristics of dendritic cells (DCs) derived from peripheral blood mononuclear cells (PBMCs) of patients diagnosed with syphilis. Methods PBMCs were isolated from 16 patients clinically and serologically diagnosed with syphilis, and from 16 healthy human controls, then cultured with GM-CSF and IL-4. On day 10, the monocyte-derived dendritic cells (MoDCs)of the patients and controls were collected and subjected to the detection of surface molecules by flow cytometry; TpN17 was used to stimulate MoDCs from the controls, the expression of phosphorylated ERK was detected by Westem blotting 20 minutes following the stimulation. Results The positivity rate of CD80 was significantly increased in the patients with syphilis than that in the controls (51.90% vs 33.67,P ＜ 0.05), while no significant difference was observed in the expressions of CD83, CD86 or HLA-DR be tween the two groups (16.53% vs 15.99%, 66.13% vs 59.32%, 91.29% vs 90.51%, all P 0.05). The ex pressions of CD80 and CD83 on the surface of MoDCs were enhanced in a dose-dependent manner after ex-posure to TpN17. The expression of cytoplasmic phosphorylated ERK was observed in MoDC stimulated by TpN17, but not in those without the treatment. Conclusions Antigenic stimulation with Treponema pal-lidum may be a reason for phenotypic abnormality of MoDCs derived from patients with syphilis. TpN17 may stimulate the maturation of DCs through the ERK signal transduction pathway.

Aim To investigate the effects of monocrotaline pyrrole on production of nitric oxide and expression of eNOS protein of cultured pulmonary artery endothelial cells and on contractility of cultured pulmonary artery smooth muscle cells.Methods DAF-2 fluorescence technique was used to determine NO level,Western blot analysis was performed to determine the level of eNOS protein,and collagen gel contraction system was adopted to analyze muscle contractility.Results NO production induced by ACh and expression of eNOS protein were obviously inhibited by monocrotaline pyrrole compared with those of control group and gel contraction area in MCTP-treated cells induced by Thapsigargin obviously decreased.Conclusions monocrotaline pyrrole could inhibit the level of the ACh-induced production of NO and expression of eNOS protein,and enhance the contractility of pulmonary artery smooth muscle cells,which may be one of the possible mechanisms of MCTP-induced pulmonary artery hypertension.

Clinical immunology should display the advanced and applied characteristics of immunology,and show the speciality of clinical practice.This course is a bridge between fundamental immunology and clinical teaching so that the medicos should get better foundation for the future learning.

Objective To measure the organ weights , blood physiological and biochemical parameters , and immune function of Specific Pathogen Free ( SPF) Rag2 knockout ( KO) mice.Methods Rag2 knockout mice were selected at five and ten weeks , and the organ weights , blood physiological and biochemical parameters were observed .The percentages of CD3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were checked by FCM in Rag2 KO mice at six week of age in terms of its T , B lymphocyte function and NK cell activity .Results Among the same sex group of NOD/SCID mice, the weights of brain, lung, spleen, liver, heart, kidney, and the levels of TBIL, WBC in 10 weeks of age are higher than 5 weeks.At the same age, the weights of heart, kidney, liver, spleen, and the levels of AST, ALP, A/G, GLU, PLT, PCT, WBC, and LYM%in male mice are higher than females.The Rag2 KO mice lacks T cells(0.36 ±0.15)%、CD4+T cells(0.21 ±0.06)%、CD8+T cells(0.23 ±0.07)%、CD19+B cells(0.28 ±0.04)%、B220+B cells(2.03 ±0.42)%).The percentage of NK cells is(24.13 ±3.62)%, and the percentage of granulocytes is (57.20 ±3.85)%.Conclusion The study suggests that the organ weights , blood physiological and biochemical parameters are affected by age and gender in Rag2 KO mice, which main biological characteristics are similar with C 57BL/6J mice.The Rag2 KO mice show the deficiency of T , B cells function .

BACKGROUND:Changes of both stem cell markers and endothelial cell phenotype help understand characteristics of endothelial progenitor cells during adherent differentiation.However,there is still no specific cell marker to distinguish from mature endothelial cells.OBJECTIVE:To study the changes of stem cell markers and endothelial cell phenotype during the differentiation of rat peripheral blood rnononuclear cells into endothelial cells.DESIGN.TIME AND SETTING:Cell observation study was performed in the Laboratory of Cardiac Surgery,Qingdao Municipal Hospital between June 2004 and December 2008. MATERIALS:Peripheral blood was drawn from male SD rats to obtain mononuclear cells by Ficell density gradient centrifugation. METHODS:Mononuclear cells were in vitro cultured in fibronectin culture medium and induced by vascular endothelial growth factors(VEGF)and basic fibroblast growth factors(bFGF)in order to stimulate a differentiation into endothelial cells. MAIN OUTCOME MEASURES:Adherent cells in the culture system were identified for CD31,CD34,Rk-1 and vWF with immunochemistry within 1-7 days.RESULTS:The expressions of CD31.CD34,FIk-1.vWF on adherent cells were different in different time durations.The expressions of CD31 and CD34 started on the 2nd day of culture.reached the peak on the 4th day,gradually decreased and even disappeared on the 6thday.While.FIk-1 expressed on the 3rd day of culture,gradually increased,and reached at the peak on the 7th day.vWF expressed gradually until 100%on the 7th day. CONCLUSION:The differentiation of peripheral blood stern cells into endothelial progenitor cells is characterized by the appearance of endothelial cell phenotypes and the disappearance of stern cell markers.both in the manner of gradual progression.

Objective To investigate the synthesis of heat shock protein 70 (HSP70) induced by norepinephrine preconditioning on donor heart and its effects on myocardial cell apoptosis and apoptosis related proteins. Methods 18 Wistar rats were random divided into 2 groups, with 9 in each group. The rats in the control group were intraperitoneally injected with 0.5 ml saline. After 24 hours, hearts were isolated and stored with histidine-tryptophan-ketoglutarate (HTK) solution at 4 ℃ for 3 hours to establish Langendorff isolated heart models, and then isolated hearts were perfused by Langendorff model with Krebs-Hense leit (K-H) solution for 2 hours. The rats in the experimental group received intraperitoneally 3. 1 μmol/kg (0. 53 mg/kg) noradrenaline bitartrate that was dissolved in saline and hearts were isolated and stored after 24 hours. Followed process was the same as that in the control group. Myocardial HSP70, Bcl-2, Bax content, apoptosis index were measured, cell structures were observed under light and electron microscope.Results HSP70 in the experimental group were higher [(17.78 ± 1.82)%] than those in control group [(5.22 ± 1.05)%], and biochemical indicators in texperimental group[(41.88 ± 5.09)%, (22.61 ±3. 49 ) %] were better than those in control group [(31.36 ± 3. 27 ) %, ( 40. 52 ± 4. 1 7) %]. There were alleviated ultrastructure injures in experimental group compared with those in control group. Conclusions This study demonstrated that norepinephrine preconditioning could induce high expression of HSP70 and it could play a very important role during ischemia-reperfusion. It could protect the structure and function of myocytes in isolated rat hearts and inhibited myocardial apoptosis.

AIM: To solve the two difficulties of bone resorption and inflammation in rheumatoid arthritis, clone the recombinant human osteoprotegerin (OPG) and mycobacteria heat shock protein 70 (HSP70) functional gene,and study the expression and activity of OPG-HSP70 fusion protein in E.coli. METHODS: Experiments were performed at the Laboratory of Department of Immunology, Capital Medical University from May 2006 to September 2007. Complementary DNA encoding full length OPG protein was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from human osteosarcoma cell line MG63 and cloned into pGEMT-Easy vectors. Then, using the recombinant plasmid as the template,the DNA encoding the fusion protein OPG-HSP 70 was amplified by PCR,and was inserted into prokaryotic expression vector pET-28a. Construct pET-28a-OPG-HSP 70 was used to transform into competent E.coli. BL21(DE3) which were induced by isopropyl B-D-thiogalactopyranoside (IPTG),and the fusion protein from above E.coli was collected. Sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western-blotting were performed to identify OPG-HSP 70 fusion protein. The experiments of osteoclast inhibition and restraining inflammation were used to detect the bioactivity of fusion protein. RESULTS: ①The complementary DNA encoding full length OPG protein was obtained. OPG-HSP 70 fusion gene obtained in this experiment was successfully inserted into pET-28a vector. OPG-HSP70 fusion protein was expressed when transformed into E.coli.BL21(DE3). ②SDS-PAGE indicated that the fusion protein was large expressed at the molecular weight of Mr22 000, but there were no band in the total lysate of bacteria harboring pET-28a-OPG-HSP70 without IPTG induction group. ③Western-blotting indicated that the OPG-HSP70 fusion protein could specifically react with anti-human OPG monoclonal antibodies. ④The osteoclast inhibition test demonstrated that the fusion protein could reduce the number of osteoclast, and had the ability to inhibit bone absorptions in vitro. ⑤The experiment of restraining inflammation showed that the fusion protein could significantly reduce the inflammation of delayed type hypersensitivity (DTH) mice, which explained that HSP70 in fusion protein had the inflammation inhibitory bioactivity. CONCLUSION: OPG-HSP70 fusion protein is expressed in E.coli.BL21(DE3),and function study in vitro illustrates the bioactivity of fusion protein.

BACKGROUND: How to successfully obtain compact endothelium layers on smooth muscle cells is the most crucial part for the tissue-engineered vessels. OBJECTIVE: To explore the effects of different cell implantation concentrations on the construction of the complete biological tissue-engineered blood vessels.METHODS: Different concentrations of porcine vascular smooth muscle cells (5×105, 5×107 cells/L) were implanted on the porcine acellular vascular matrix to culture for 3 days. Then different concentrations of endothelial progenitor cells (5×105, 5×107 cells/L) were implanted on the smooth muscle cell-vascular matrix composite to construct lamellar complete biological tissue-engineered blood vessels.RESULTS AND CONCLUSION: The growth curves of high concentrations of smooth muscle cells on the acellular vascular matrix were similar to that of low concentrations. Moreover, the growth curves of cells implanted in the culture plates were similar to that implanted on the acellular matrix. However, cells in the low concentration groups have relatively low proliferation activity and low coverage rate. The cell coverage rate decreased as follows: high concentrations of endothelial progenitor cells+acellular matrix containing high concentrations of smooth muscle cells > high concentrations of endothelial progenitor cells+acellular matrix containing low concentrations of smooth muscle cells > low concentrations of endothelial progenitor cells+acellular matrix containing high concentrations of smooth muscle cells > low concentrations of endothelial progenitor cells+acellular matrix containing low concentrations of smooth muscle cells. Moreover, high concentrations of endothelial progenitor cells form relatively compact layers on the acellular matrix and show cobble-like growth. These findings indicate that an increase in the cell implantation concentrations is beneficial to the rapid formation of compact cell layers on the material surface.

Objective To study the curative effect of continuous veno-venous hemodiafiltration(CVVH)in patients with acute renal failure(ARF)after aortic dissection surgery. Methods Fifteen patients with renal dysfunction following aortic dissection surgery underwent CVVH from Feb. 2002 to Dec. 2009 in this study.The clinical data of these patients were collected,such as heart rate(HR),central vein pressure(CVP),mean artery blood pressure(MAP),PaO2,renal function,perioperative manifestations and outcomes. Results Eleven patients survived but 4 died during the course of treatment. There were significant decreases of BUN,Creatinin after CVVH (P ＜ 0. 05)treatment,and the urine volume returned to nomal after CVVH in 6 -40 days. Conclusions CVVH is an effective,convenient and safe treatment for patients with severely ARF following aortic dissection surgery.

0.8). Conclusions This DNA chip combined with multiplex PCR is a rapid diagnostic assay with high specificity and sensitivity for the detection of Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma Urealyticum and their drug-resistance, and may be applied in the diagnosis of urogenital infections.