Objective To observe the immunohistochemical changes of ICAM-1,MMP-2 and MMP-9 in the lungs of immunocompromised rats with Pseudomonas aeruginosa pneumonia and their relationships with lung inflammation.Methods After the establishment of pseudomonas aeruginosa pneumonia infected immunocompromised rat mode,the pathological changes of lungs were observed, lung wet/dry ratios and total protein concentration in bronchial alveolar lavage fluid were tested,and imunnohistochemical study of ICAM-1,MMP-2 and MMP 9 in lung tissue were performed.Results 1.The staining intensity of ICAM-1 in alveolar epithelial cells turned stronger in rats with pulmonary infection than those without of both groups(P＜0.05);2.The staining intensity of MMP-2 in lung tissue was stronger in rats with pulmonary infection than those without infection in both groups,and reached peak at 6～9 h after inoculation.Immunohistochemical changes of MMP-9 exhibited a similar pattern,4.Immunohistochemical changes of ICAM-1,MMP-2 and MMP-9 showed some correlation with numbers of polymorphonuclears in lung tissue(P＜0.05);5.A correlation between the stai- ning intensity of MMP-9 in bronchial epithelial eells and total protein concentrations were observed(r_s =0.484,P＜0.05),similar association were found between the staining intensity of MMP-2 in alveolar epithelial cells,endothelium of arterioles and venules and tissues beneath endothelium and to- tal protein eoncentrations in bronchial alveolar lavage fluid(r_s were 0.457,0.492 and 0.429,respec- tively,P＜0.05).Conclusion In immunocompromised rats,the staining intensity of ICAM-1, MMP-2 and MMP-9 in lung tissue of those with pseudomonas aeruginosa pneumonia were stronger than those without infection,and the changes were demonstrated some correlation with the levels of polymorphonuclears infiltration or severity of lung injury.

Objective To study inflammatory reaction induced by N-protein of severe acute respiratory syndrome(SARS)-coronavirus(CoV)in human alveolar typeⅡepithelial cell(A549). Methods Effects on growth of A549 cell by N-protein of SARS-CoV:activity of A549 cells was determined by thiazylyl blue colorimetry assay at 24,48,72 and 96 h,respectively.Effects on cyto- kine production by A549 cells exposed to N-protein of SARS-CoV:interleukin(IL)-6,IL-10 and transforming growth factor-?1(TGF-?1)concentration in culture supernatant were determined by enzyme-linked immunosorbent assay(ELISA).Effects on mRNA expression of cytokine of A549 cells and matrix metalloproteinases-9(MMP-9)exposed to N-protein of SARS-CoV:total RNA of A549 cells was extracted using Rneasy mini kit;RT-PCR was employed to measure the mRNA expression of IL-6,IL-10,TGF-?1 and MMP-9 semiquantitatively.Results Different concentrations of N-protein could all inhibit the growth of A549 cells(after 48 h)and the inhibition by 20?g/mL pro- tein was the strongest.Compared with the control group(0.737?0.024,0.968?0.007),the A val- ues of experimental groups at 72 h and 96 h(0.672?0.027,0.799?0.092)decreased obviously (P

A human granulocyte-macrophage colony stimulating factor (GM-CSF)/interleukin-3(IL-3) fusion gene with a short linker between the GM-CSF and IL-3 gene has been successfully constructed and expressed in E. coli under the control of T7 promoter. The recombinant fusion protein was expressed as inclusion bodies after the IPTG induction. The yield of the GM-CSF/IL-3 fusion protein was over 30 % of the total cellular proteins. Western-blotting results showed that the fusion protein could specifically combined with GM-CSF antibody and IL-3 antibody. The biological activity was detected by the GM-CSF and IL-3 dependent cell line TF-1. After solubilizing with 8mol/L urea and renaturing with dialysis against Tris. HCl solution,the refolded fusion protein showed obvious activities to maintain the growth of TF-1 cell.

Hyperglycemia, advanced glycation end products (AGEs), hyperinsulinemia and dyslipidemia may play roles in the development of diabetes-associated atherosclerosis and post-angioplasty restenosis. Clinically, their effects seem to be synergic. However, few studies have focused on the synergistic action of these factors. In the present study, we investigated whether glycated serum albumin (GSA) has a synergistic effect with insulin on the proliferation of vascular smooth muscle cells (VSMCs). VSMCs were isolated from rat thoracic aortas and cultured in fetal bovine serum (FBS)-free medium for 24 h, then exposed to GSA, insulin or GSA + insulin for 48 h with or without pretreatment of mitogen-activated protein kinase (MAPK) inhibitors or the antioxidant N-acetylcysteine (NAC). Cell growth rate was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay or cell counting. The changes of phosphorylated-p38 MAPK and phosphorylated-C-Jun N-terminal kinase 1/2 (JNK1/2) were measured by Western blot analysis. The results showed that only p38 MAPK, but not JNK was activated by GSA and insulin co-incubation. VSMC proliferation was increased by insulin (10-1000 nmol/L) or GSA (10, 100 microg/mL). Co-incubation of insulin (100 nmol/L) and GSA (100 mug/mL) caused a more potent increase in VSMC proliferation than insulin or GSA incubation alone. p38 MAPK inhibitor, SB203580, as well as NAC, could inhibit the VSMC proliferation induced by co-incubation of GSA and insulin. The results show that insulin enhances GSA-induced VSMC proliferation, which may be mediated through a reactive oxygen species (ROS)-p38 MAPK pathway. The synergism of AGEs and insulin may play a detrimental role in the pathogenesis of diabetic atherosclerosis and post-angioplasty restenosis.
Animals ; Aorta, Thoracic ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drug Synergism ; Insulin ; pharmacology ; physiology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Serum Albumin ; pharmacology ; physiology ; p38 Mitogen-Activated Protein Kinases ; metabolism

To explore a simple and effective method to determinate the volume of CD34(+) cells in the peripheral blood of donors received drug mobilization for stem cell transplantation by using flow cytometry, the mobilized peripheral blood from donors and 100 micro l fresh whole blood were labeled with monoclonal antibodies Anti-CD34-PE and Anti-CD45-FITC, after lying the red blood cells, and assessed with flow cytometer FL2 (log) vs SSC (log) and FL1 (log) vs SSC (log) were mainly used for analysis windows. The results showed that a level of CD34(+) cells in whole nucleated cells as low as 0.05% - 0.1% can be detected effectively using this method when 10(5) nucleated cells were counted. At day 5 or day 6, the level of CD34(+) cells in most samples of patients reached a peak volume, some of samples and the levels were more than one percent in. It was concluded that CD34(+) cells can be effectively determined by using this method. According to the relative rate of CD34(+) cells, the time to harvest the stem cells in blood can be determined.
Antigens, CD34 ; blood ; Blood Donors ; Flow Cytometry ; methods ; Hematopoietic Stem Cells ; cytology ; Humans

BACKGROUNDThe number of immunosuppressed patients has increased in the past decades. Among them Pseudomonas aeruginosa (P. aeruginosa) is one of the leading bacteria for pneumonia that are associated with poor prognosis. However, the pathogenesis of P. aeruginosa pneumonia in immunosuppressed patients is not understood completely. Previous reports showed keratinocyte growth factor (KGF) is associated with lung injury in immunocompetent hosts. In this study, we investigated the different reactions of lung injury, lung pathology and KGF expressions in P. aeruginosa pneumonia between immunosuppressed and immunocompetent rats.

METHODSImmunosuppression of male rats was induced by injecting immunosuppressive subcutaneously. Pneumonia was established by instilling P. aeruginous tracheally. The immunocompetent rats were the control group. Survival rate, lung histopathology, pulmonary permeability and oedema, KGF mRNA and protein expressions in lungs of both groups were investigated.

RESULTSThe survival rate of immunosuppressed group was lower than that of immunocompetent group (33.3% vs 83.3%). After exposure to bacteria, pulmonary permeability and wet/dry ratio in immunosuppressed group were higher than those in immunocompetent group. Pulmonary congestion and haemorrhage were more intensive in immunosuppressed group compared to immunocompetent group. Apoptosis and necrosis were also observed in infected lungs of immunosuppressed rats. Although we detected KGF expressions in lungs of both groups after infection, the expressions of KGF protein and mRNA gene in immunosuppressed group were much lower than in immunocompetent group.

CONCLUSIONSCompared with immunocompetent group, there was more intensive lung injury in immunosuppressed group. Severe lung injury may contribute to the poor prognosis of pneumonia. KGF expressions of pneumonia in immunosuppressed rats were less than those in immunocompetent ones.

Animals ; Capillary Permeability ; Fibroblast Growth Factor 7 ; analysis ; genetics ; Immune Tolerance ; Leukocyte Count ; Lung ; metabolism ; pathology ; Male ; Pneumonia, Bacterial ; metabolism ; Pseudomonas Infections ; metabolism ; mortality ; pathology ; Pulmonary Edema ; etiology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Survival Rate ; Up-Regulation

Objective To compare the efficacy and safety between cryoablation (Cryo) and radiofrequency (RF) ablation for treating patients with atrioventricular nodal reentrant tachycardia (AVNRT). Methods Patients with AVNRT (n=304) were divided into Cryo group (n=67) and RF group (n=237). The procedure success rate, complete slow pathway block rate, atrioventricular block rate and relapse rate were compared between two groups. Results There was no statistically difference between 2 groups in the success rate (Cryo group 98.5% vs RF group 97.0%, P=0.820), complete slow pathway block rate (Cryo group 98.5% vs RF group 91.6%, P=0.088), atrioventricular block rate (Cryo group 0 vs RF group 2.5%, P=0.413), relapse rate (Cryo group 0 vs RF group 1.7%, P=0.643). But Cryo group had more advantage than RF group. Conclusion Efficacy and safety were comparable between cryoablation and radiofrequency ablation for treating patients with AVNRT.

Objective To explore the feasibility and methodology of radiofrequency catheter ablation (RFCA) guided by 3D navigation system (Ensite-NavX) for right atrioventricular accessory pathway.Method Thirty-three cases of right accessory pathway atrioventricular reentrant tachycardia including 16 cases in right free wall,3 in right middle septum,14 in right posterior septum; 23 cases of dominant accessory pathway and 10 cases of concealed were treated by RFCA guided by NavX navigation.NavX navigation modeling method or spatial localization method was exploited to locate target positioning.Result All patients were successfully ablated without serious complications.Among them,25 cases were operated without exposure to X-ray,7 patients were exposed for several seconds to verify catheter position,1 case in right free wall was ablated under X-ray combined with Swartz sheath ablation.Conclusion Nonfluoroscopy or less fluoroscopy RFCA for right atrioventricular accessory pathway with Ensite-NavX is safe and feasible,modeling or spatial orientation method are helpful to locate the ablation target positioning.

BACKGROUNDAzithromycin can reduce neutrophil accumulation in neutrophilic pulmonary diseases. However, the precise mechanism behind this action remains unknown. Our experiment assessed whether azithromycin inhibits neutrophil accumulation in the airways by affecting interleukin-17 (IL-17) downstream signals.

METHODSMice were pretreated with azithromycin before murine IL-17A (mIL-17) stimulation. After the mIL-17 stimulation, the levels of six neutrophil-mobilizing cytokines were determined by enzyme-linked immunosorbent assay (ELISA) tests in bronchoalveolar lavage (BAL) fluid; IL-6, CXC chemokine ligand-1 (CXCL-1), CXCL-5, macrophage inflammatory protein-2 (MIP-2), granulocyte colony-stimulating factor (G-CSF), and granulocyte macrophage colony-stimulating factor (GM-CSF). The number of neutrophils in BAL fluid were evaluated by cytospin preparations.

RESULTS(1) Azithromycin pretreatment significantly inhibited both the release of three neutrophil-mobilizing cytokines (MIP-2, CXCL-5 and GM-CSF) and the accumulation of neutrophils in airways caused by mIL-17 stimulation. (2) The levels of three neutrophil-mobilizing cytokines (IL-6, MIP-2 and GM-CSF) were positively correlated with the numbers of neutrophil in BAL fluid.

CONCLUSIONSAzithromycin can inhibit neutrophil accumulation in the airways by affecting IL-17 downstream signals. This finding suggests that macrolide antibiotic application might be useful in prevention of neutrophilic pulmonary diseases characterized by high levels of IL-17.

Animals ; Azithromycin ; pharmacology ; Bronchoalveolar Lavage Fluid ; chemistry ; Chemokine CXCL2 ; metabolism ; Chemokines, CXC ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Granulocyte Colony-Stimulating Factor ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; metabolism ; Interleukin-17 ; pharmacology ; Interleukin-6 ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Neutrophils ; drug effects ; metabolism

BACKGROUNDThe down-regulation of constitutive nitric oxide synthase (cNOS) and up-regulation of inducible nitric oxide synthase (iNOS) are associated with the allergen-provocated airway hyperresponsiveness (AHR). This study aimed to determine whether their alteration also plays an important role in the AHR induced by lipopolysaccharide (LPS).

METHODSHartley male guinea pigs, weighing between 250 g and 350 g, were injected with LPS at a dose of 1 mg/kg every 24 hours for three days. A non-selective NOS inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME), or a selective inducible NOS inhibitor, aminoguanidine (AG), were used thirty minutes before each injection of LPS. Airway reactions, nitric oxide (NO) production and inflammatory changes were detected 24 hours after the last dose of LPS.

RESULTSAG significantly decreased the NO production in the bronchoalveolar lavage fluid (BALF) and sharply reduced the intensity of bronchoconstriction to histamine challenge. L-NAME also significantly decreased the NO production in the BALF, but had no effect on airway reactions or, perhaps, a tendency to enhance the intensity of AHR.

CONCLUSIONSThe data suggest that inducible NOS contributes to the AHR induced by repetitive intraperitoneal LPS, and constitutive NOS was also involved.

Airway Resistance ; drug effects ; Animals ; Bronchial Hyperreactivity ; chemically induced ; Enzyme Inhibitors ; pharmacology ; Guanidines ; pharmacology ; Guinea Pigs ; Lipopolysaccharides ; toxicity ; Male ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; antagonists & inhibitors ; physiology