Objective To construct the replication-incompetent lentivirus expressing rat TLR4 shRNA,and to observe its inhibitory effect on TLR4 expression in alveolar macrophage and on lipopolysaccharide (LPS)-induced release of IL-1?,IL-6.Methods We designed and constructed four shRNA expressing plasmids with silencing effect,then they were co-transfected into HEK293T cells with the TLR4 eukaryotic vector constructed previously;the best shRNA was selected to be packed into lentivirus;and the titer was determined.The packed lentvirus was used to infect the alveolar macrophage(NR8383) in presence of LPS,and the LPS-induced IL1?,IL-6 expression was examined by ELISA.Results The best shRNA was successfully screened out and was correctly inserted into the lentivirus.The titer of the recombinant lentivirus was 2.0 ? 106 TU/ml.The LPS-induced expression of IL-1?,IL-6 in the alveolar macrophages was greatly reduced after virus infection (P

Th17 cells play an important role in the pathogenesis of various immune-mediated diseases,including psoriasis,rheumatoid arthritis,multiple sclerosis,inflammatory bowel disease,asthma,and so on.The discovery of Th17 cells has offered scientists a new insight into the etiology and treatment of a broad spectrum of diseases.This article presents an overview of the differentiation,cytokine expression and trafficking of Th17 cells.

Objective To construct eukaryotic expressed plasmid vector pLenti6/V5- DEST-Cx43 in human for further study on bioinformative communication between myocardial cell, embryonic development of heart and differentiation of myocardial cell. Methods Cx43 cDNA fragments were obtained from plasmid p18-T by restriction enzyme BamH1、Xho1, after the agarose gel electrophoresis was performed, the Cx43 cDNA was retrieved. pENTR11 were extracted by enzyme BamH1、Xho1, then connected with Cx43 cDNA by T4 DNA ligase. LR reaction was performed and Cx43 cDNA were cloned into Lentiviral vector pLenti6/V5-DEST. Results Agarose electrophoresis and sequent examination, identified that Cx43 cDNA was cloned into Lentiviral vector pLenti6/V5-DEST. Conclusion The present experiment demonstrate a successful cloning of human Cx43 cDNA into the Lentiviral vector pLenti6/V5-DEST.

Objective To summarize the experience of heart-lung transplantation.Methods Four patients with Eisenmenger’s syndrome underwent heart-lung homoplastic transplantation. All patients were complicated with severe pulmonary hypertension in New York Heart Association ( NYHA ) functional class IV. Cannulation for cardiopulmonary bypass consisted of a cannula in the high ascending aorta and separate vena caval cannulas. The heart-lung graft was moved into the chest, beginning with passage of the lung before the phrenic nerve pedicle. The bronchus was trimmed, leaving two cartilaginous rings proximal to the orifice of the upper lobe. The tracheal anastomosis was performed with a continuous 4-0 polypropylene suture, with the posterior portion continuously and anterior interrupted. The lungs were then ventilated (

The most important cause of late mortality after lung transplantation is obliterative bronchiolitis ( OB) . It is clear that a good animal model is indispensable to further unravel and clarify the pathogenesis of bronchiolitis obliterans syndrome ( BOS) .Many animal models have been developed to study BOS, however, so far, none of these models truly mimics the human condition.In recent years mouse models of orthotopic lung transplantation have been established, which provide potential possibilities for further studies of OB/BOS after lung transplantation.The aim of this article was to review the pros and cons of those animal models, and discuss the possible approaches to establish animal models of chronic rejec-tion after lung transplantation.

Objective To determine the prevalence of latex glove(LG)-induced type Ⅳ hypersensitivity in clinical nurses. Methods 100 clinical nurses were selected,among whom 69 nurses were set as the latex glove hypersensitivity group,and the other 31 nurses did not show hypersensitivity to latex glove (the control group).The two groupe underwent a patch testing with a modified European standard series of allergens supplied by Chemotechnique Diagnostics.The positive allergens were compared between the two groups. Results The positive rate of patch testing in patients of the latex glove hypersensitivity group was superior to that in the control group (73.9％ vs 25.8％).Cobalt chloride and potassium dichromate were the most common allergens in both groups,while only formaldehyde and para-phenylenediamine were more susceptible in the latex glove hypersensitivity group compared with that in the control group.26.1％ of the latex glove hypersensitivity group was responsive to rubber additives,but in the control group the results were negative.Conclusions The hand dermatitis of female nurses with LG allergy is mainly caused by exposure to daily necessities.The prevalence of LG-induced type Ⅳ hypenensitivity is relatively high in clinical murses.

Objective To investigate the prevalence and clinical characteristics of thyroid dysfunction induced by IFNα therapy in patients with chronic hepatitis.Methods Thyroid function of 310 patients with chronic hepatitis receiving IFNα therapy were evaluated.Results Serum free tri-iodothymnine (F13),free thyroxine(FT4)and sensitive thyroid-stimulating hormone(sTSH)of all patients were normal and TGAb was negative before IFNα therapy.After the treatment,10 patients(10/3 10,3.22%)had thyroid dysfunction,in which 7 were of hypothyroidism and 3 were of hyperthyroidism.The thyroid function of all patients recovered within 1-year follow-up.Conclusions IFNα therapy may induce thyroid dysfunction in patients with chronic hepatitis,in which prognosis can be good when appropriate treatment is given.

Objective This study evaluated the effects of combined procedure of surgical treatment and cell transplantation on dilated cardiomyopathy. Methods Eight patients (5 men and 3 women) with moderate to severe mitral regurgitation from end-stage dilated cardiomyopathy underwent surgery. Four patients were in functional class (FC) Ⅳ, six were in FC Ⅲ. There age ranged from 15 to 56 years. The preoperative ejection fraction (EF) ranged from 0.15 to 0.32 (mean 0.26 ± 0.08). Mitral valve replacement was performed in 5 patients and mitral valve repair in 3. Auto-bone marrow monenuclear cells were harvested, isolated, washed, and resuspended for direct injection after surgical procedure. Results All patients survived and were discharged from the hospital. After a mean follow-up period of 18 months ( 12 - 42 months). Echocardiography showed postoperative ejection fraction and wall movement velocity increased after 6 months. Radionuclide ventriculography showed myocardial perfusion improved significantly. Conclusion Combined procedure of surgical treatment and cell transplantation led to significant improvement in cardiac function in patients with dilated cardiomyopathy.

Objective:To establish a rapid method to detect drug-resistance genotypes of extended spectrum-β-Lactamases (ESBLs) produced by gram negative bacillus using the real -time fluorescence quantitative PCR. Methods: According to clinical common genotypes of ESBLs, SHV, TEM.CTX-M.OXA and their homology, 9 pairs of specific primers were designed including SHV, TEM, CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, OXA-1, OXA-2 and OXA-10. To extract DNA template by boiling assay, and then establish and grade up SYBR GREEN I real-time fluorescence quantitative PCR reaction system, finally definite real-time fluorescence quantitative PCR method. Its precision and range of linearity were tested. With established assay 51 multi- drug resistant ESBLs- E. coli K. pneumoniae were detected and compared with improved three dimensional extract tests. Results: Except OXA-2, 8 genotypes SHV, TEM, CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, OXA-1 and OXA-10 were amplified by quantitative PCR from 39 ESBLs+ and 51 multi-drug resistant ESBLs-E. coli K. pneumoniae and confirmed by sequence testing. The range of linearity was 3×10~3-3×10~8 copies/mL, r =-0.994 7. Repetitive experiments showed that the average coefficient of variation between -runs was 9.6%. Comparing with three dimensional extract test, there was no significant difference (χ2 = 1.125,P> 0.05). Conclusion: Testing drug-resistance genotypes of ESBLs with SYBR GREEN I real-time fluorescence quantitative PCR is a rapid,specific and sensitive method, which is capable of inspecting genotypes of ESBLs from clinical strains.

Objective To explore the electrophysiological properties and the disparity of visual cortical neurons in developing rats. Methods Whole cell patch clamp recording and intracellular labeling of brain slices were performed on rats at postnatal 14 and 28 days. The electrophysiological data were analyzed according to the cellular input resistance and resting membrane potential. Results When the input impedance of visual cortical neurons(IR) was lower, increased peak value of postsynaptic currents(PSCs), prolonged rise and decay time were found. At postnatal 14th day(before eyes opened), the intermediate cells and mature cells were 59.2% and 14.9% respectively, but at postnatal 28th day(14 days after eyes opened), the mature cells and immature cells were 62.5% and 12.5% respectively. Conclusion During the period of postnatal development, the visual cortical neurons become mature gradually, but maturation is not completely correlated with age.