Objective To assess the clinical significance of three different noninvasive airway inflammatory indices in induced sputum and exhaled breath condensate ( EBC ) from persistent asthmatic patients.Methods Moderate and severe asthmatic patients were prescribed inhaled corticosteroids combined with long-acting β2 agonists for a month.The symptom scores and percentage of predicted value of forced expiratory volume in one second ( FEV1 ) ( FEV1% pred) were measured while the concentrations of H2O2,NO3-/NO2-,and cysteinyl-leukotriene E4(LTE4) in induced sputum and EBC were detected before and after therapy.Results A total of twenty-five subjects with moderate and severe asthma were enrolled.By combined therapy for one month the asthma aymptoms relieved and FEV1% pred improved significantly (P<0.01).The concentrations of H2O2,NO3-/NO2- arid LTE4in induced sputum and EBC declined significantly(P<0.01) although the concentrations were still higher than those at normal baseline.More marked reduction of H2O2 and NO3-/NO2- compared to LTE4was observed.It was revealed that the concentrations of H2O2 and NO3-/NO2- but not of LTE4in EBC were negatively correlated with FEV1% pred (P <0.01) and positively with symptom scores.Such correlations were also found in H2O2 in induced sputum with FEV1 % pred and symptom scores as well as NO3-/NO2- in induced sputum with FEV1 % pred.The improvement of FEV1 % pred after treatment was positively correlated with the reduction of H2O2 and NO3-/NO2- both in induced sputum and EBC.Correlation analysis also demonstrated three inflammatory indices were equivalent in induced sputum and EBC( correlation coefficient of H2O2,NO3-/NO2- and LTE4,0.759,0.826 and 0.653,respectively.P < 0.01 ).Conclusions (1) Combined therapy with inhaled corticosteroid plus long-acting β2 agonist significantly improves the clinical symptoms and lung function of patients with moderate and severe asthma companied with marked suppression of airway inflammation.(2) Both of EBC and induced sputum sampling are valuable noninvasive procedures for detecting asthma airway inflammation,however,EBC technique is superior in safety and reproducibility.(3) H2O2 and NO3-/NO2- seem to be more sensitive indices in diagnosis and monitoring asthma compared to LTE4.

Objective To study the DNA damage in hippocampal cells of rats induced by 1 800 MHz electromagnetic irradiation. Methods Wistar rats were randomly divided into four groups, 2 exposed groups and 2 controls. The experimental subjects were exposed to the 1 800 MHz (continuous wave, CW) electromagnetic fields for 21days (12 h/d), the power densities were 0.5 mW/cm2 and 1.0 mW/cm2, meanwhile, sham-control exposures were carried out, the DNA damage was examined by Comet assay. Results The tailed rate and the tail area of the hippocampal cells in 1.0 mW/cm2 group was higher or larger than those in the control and 0.5 mW/cm2 groups(P

Objective: To investigate the role of miR-455 in cardiac hypertrophy with its potential cellular and molecular mechanism in mice. Methods: The mice model of cardiac hypertrophy was established by transverse aorta constriction (TAC), and 18 male kunming TAC mice were randomly divided into 3 groups:①TAC + miR-455 group,②TAC + GFP (green lfuorescence protein) group and③Sham group (sham operation + GFP).n=6 in each group and all animals were treated for 2 weeks. The hemodynamic and echocardiographic indexes were examined, histo-pathological changes of myocardial tissue were observed by HE and Masson staining. The hypertrophic and ifbrosis gene expressions were measured by RT-PCR, the apoptosis protein level was detected by Western blot analysis. The expressions of miR-455 targeting gene and protein were also determined. Results: Upon 2 weeks modeling, compared with Sham group, TAC+GFP group had increased ratio of heart weight/ body weight (9.78 ± 0.20) mg/g vs (8.25 ± 0.22) mg/g,P<0.01, increased left ventricular diastolic (LVD) wall thicknesses (1.782 ± 0.058) mm vs (1.457 ± 0.050) mm,P<0.05, decreased LVD diameter (3.027 ± 0.052) mm vs (3.142±0.050) mm,P<0.05, increased LVEF (84.167 ± 4.167) % vs (77.000 ± 3.347) %,P<0.05; increased gene expressions of cardiac hypertrophy and ifbrosis, allP<0.05, decreased anti-apoptosis protein and increased promoting apoptosis protein, allP<0.05. Compared with TAC+GFP group, TAC+miR-455 group presented increased ratio of heart weight/body weight (12.04 ± 0.11) mg/g vs (9.78 ± 0.20) mg/g,P<0.01, increased LVD wall thicknesses (1.908 ± 0.062) mm vs (1.782 ± 0.058) mm,P<0.01, decreased LVD diameter (2.893 ± 0.069) mm vs (3.027 ± 0.052) mm,P<0.01, while LVEF was similar between 2 groups,P>0.05; increased gene expression of cardiac hypertrophy,P<0.05, while gene expression of cardiac ifbrosis was similar between 2 groups,P>0.05; the anti-apoptosis protein and promoting apoptosis protein expressions were similar between 2 groups. Compared with Sham group, TAC+GFP group had increased expressions of calreticulin (CALR) and glucose-regulated protein 78 (GRP78), allP<0.01. Compared with TAC+GFP group, TAC+miR-455 group had decreased mRNA and protein expressions of CALR, bothP<0.01, while GRP78 protein expression was similar between 2 groups,P>0.05. Conclusion: miR-455 may promote cardiac hypertrophy induced by short-term overload pressure via targeting CALR in experimental mice.

Objective To investigate the effect of early application of aldosterone receptor blockade-spironolactone on cardiac remodeling in patients with acute myocardial infarction.Methods A total of 616 patients were enrolled in this prospective study,who were admitted to the CCU Division of the First Affiliated Hospital of Harbin Medical University from January 2008 to January 2010.Inclusive criteria were as follows:First-onset STEMI according to the diagnostic criteria in ST-segment elevation acutemyocardial infarction diagnosis and treatment guidelines and admitted within 24 hours from onset.Exclusive criteria were as follows:Non-STEMI,only right ventricular infarction and old myocardial infarction; cardiac function Killip Class Ⅳ or hypotensive state; renal dysfunction(serum creatinine ＞ 221 μmol / L); serum potassium ＞5.0 mmol / L; longer than 24 hours from onset and older than 75 years of age.All the patients were randomly divided into a control group of standard therapy and a treatment group of standard therapy combined with spironolactone 20 mg per day.Totally 528 patients were observed completely,including 266 patients in the control group and 262 patients in the treatment group.There was no statistical difference of clinical characteristics between the two groups,including age,gender,past medical history,admission situation,and treatment(P ＞ 0.05).The effect of spironolactone on cardiac remodeling、left ventricular function、renal function and blood levels of potassium were evaluated by detecting echocardiography,serum potassium and serum creatinine at one month and one year follow-up.Results Compared with the control group,the echocardiography indicators such as LVESD,LVEDD,LVEF,LAD-ML and LAD-SI were significantly improved in treatment group at one year(P ＜ 0.05).In treatment group,LVESD,LVEDD,LVPWT,LVEF,LAD-ML and LAD-SI were significantly improved at one year than one month(P ＜ 0.05,and P =0.007 to LVEF),and LVEF was significantly improved at one year than one month in control group(P =0.0277).There were no statistical differences of serum potassium and serum creatinine between the two groups.Conclusions On the basis of conventional treatment,the early combination of low dose spironolactone(20 mg/d)could inhibit cardiac remodeling at late stage and prevent heart failure.

The serum samples and corresponding cervical swabs were collected fi'om 50 women with genital warts from Tianjin city,China.The neutralizing antibodies against HPV-16,-18,-58,-45,-6 and-11 in serum samples were tested by using pseudovirus-based neutralization assays and HPV DNAs in cervical swabs were also tested by using a typing kit that can detect 21 types of HPV.The results revealed that 36%(18/50)of sera were positive for type-specific neutralizing antibodies with a titer range of 160-2560,of which 22%(11/50),12%(6/50),10%(5/50),4%(2/50),4%(2/50)and 2%(1/50)were against HPVs-6,-16,-18,-58,.45 and-11,respectively.Additionally,60%(30/50)of samples were HPV DNA-positive,in which the most common types detected were HPV-68(18%),HPV-16(14%),HPV-58(12%),HPV-33(8%)and HPV-6,HPV-11,HPV-18 and HPV-52(6% each).The concordance between HPV DNA and corresponding neutralizing antibodies was 56%(28/50)with a significant difference(P＜0.05).The full-length sequences of five HPV types(HPV-42,-52,-53,-58 and-68)were determined and exhibited 98%-100% identities with their reported genomes.The present data may have utility for investigating the natural history of HPV infection and promote the development of HPV vaccines.

Objective To study the influence of site-directed deglycosylation of the HIV-1 envelope (Env) on its immunogenicity and assembly of functional pseudovirus. Methods Site-directed deglycosylation were performed using cycling mutagenesis and selection of mutants with DpnⅠ. Single-cycle infection assay was employed to analyze the effect of the mutations on the ability of functional pseudovirus assembly. The influence of deglycosylations on the immunodeficiency of Env was evaluated using pseudovirusbased neutralization assay and ELISPOT assay. Results Mutant N197Q induced higher neutralization activities for both pseudoviruses, but lower Env-specific T-cell response. And N197Q rendered the Env to lose the ability of functional pseudovirus assembly. Mutant G2 induced higher neutralization activities for pseudovirus 74-2 but lower for pseudovirus Wt, and had almost no influence on Env-specific T-cell response and functional pseudovirus forming. Conclusion The site-directed deglycosylation of the HIV-1 Env affects the pseudovirus forming and its immunogenicity.

Objective To explore the feasibility and efficacy of endoscopic-assisted thyroidectomy (EAT) in the treatment of benign thyroid nodules.Methods Sixty-five patients with one-side benign thyroid nodules were treated with EAT (EAT group,n =33) or conventional open thyroidectomy(control group,n =32) from Jan.To Dec.2013.The operation time,blood loss amount,volume of drainage,visual analog scores (VAS) for pain severity,occurrence of postoperative complications,evaluation of cosmetic,and hospital expense were recorded.Results The operation were performed successfully in patients of two groups.Blood loss volume was significantly less in EAT group (26.7 ± 23.1) ml than in control group ((45.2-± 28.9) ml,t =2.85,P ＜0.05).The volume of drainage in EAT group was (31.4 ± 8.1) ml,less than that in control group((83.6 ± 17.3) ml,t =15.66,P ＜ 0.05).The VAS at the day of 1 st,3rd after operation were (1.5-± 0.4) and (1.0 ±0.2) in EAT group was significantly lower than those in control group ((2.2 ±0.5) and (1.5 ±0.3) ;t =6.01,7.29;P ＜0.05).Satisfaction rate of cosmetic in EAT group was higher than control group (97.0％ vs 68.8％,x2 =9.02,P ＜ 0.05).Conclusion EAT is proved with safe and feasible in the treatment of benign thyroid nodules,which is with the advantage of minimally invasive and cosmetic results.

ObjectiveTo explore the role of β-catenin in the proinvasive consequences of hypoxia in hepatocellular carcinoma (HCC).MethodsWe established in vitro and in vivo hypoxic models using the highly metastatic MHCC97 and the stable red fluorescent protein-expressing MHCC97-R cells.The role of β-catenin in hypoxia-mediated aggressiveness was investigated by β-catenin knockdown.ResultsHypoxia caused a pronounced arrest of proliferation in MHCC97 cells,suppressed tumor growth in MHCC97-R xenografts,but promoted in vitro invasiveness and in vivo metastasis.β-Catenin-silencing by short hairpin significantly inhibited the enhanced invasiveness of MHCC97 cells due to hypoxia,reduced the increase in distant metastasis by hepatic arterial ligation,but failed to further restrain cell proliferation.Conclusionβ-Catenin in HCC cells plays an essential role in the hypoxia-induced metastatic potential.A reduction of βcatenin expression inhibited the proinvasive consequences of hypoxia in HCC.

Objective To detect the protection induced by HPV-58 L2 11-200 AA in animal, and analyze the relationship between antibody or neutralizing antibody titers and the protection generated by the immunizmg agent. Methods The peptide of HPV-58 L2 11-200 AA was expressed in E. coli and the mice were immunized with the peptide after purification and adsorption with aluminum adjuvant. The protection provided by different immunizing doses was detected in the mouse model against the challenge of the pseud-ovirions of human papiilomavirus types 58. The total antibodies and neutralizing antibody titers of serum were tested with ELISA and neutralization assay against HPV-58 pseudovirus, respectively. The total antibodies or neutralizing antibody titers that can protect the mouse from infection were analyzed. Results The mice can be protected from the challenge with HPV pseudovirus when the immunizing dose was 8 μg. The neutralizing antibody can not be detected in the immune serum by neutralization assay against pseudovirus. The total anti-body level has a corresponding relationship with the protection showed in mouse model. The results of total antibodies detected by ELISA showed that when the titer of total antibodies was ≥25 000, luminescent signal can not be detected and the mice can be protected from pseudovirus infection. Conclusion HPV-58 L2 11-200 AA peptide can protect mice from pseudovirus infection. L2 peptide has a promising perspective to be a candidate vaccine and the level of total antibodies in the immune serum can be used as a surrogate for the evaluation of protection against HPV infection.

Objective To establish the mouse model for human papillomavirus types 16, 45 and 58 by the corresponding pseudovirions. Methods The 293FT cells were co-transfected with eodon-modified HPV eapsids genes together with a reporter plasmid containing the luciferase gene. The cells were collected and lysed, then the pseudovirus was collected and the titration was performed. The mouse was subcutaneous-ly injected with Depo-Provera. After 4 d and intravaginally injected with nonoxynol-9, and 6 h later pseud-oviruses were inoculated in intravaginal. After 7 d, the mouse was instilled luciferin substrate intravaginally, and the expression level of the lueiferase gene was detected by the in vivo Imaging System (IVIS). Results Three types (HPV16, HPV45 and HPV58) of pseudoviruses had been produced and the titer was 3.7×108 TU/ml, 1.5×108 TU/ml and 1.2×108 TU/ml, respectively. The luminescent regions could be detected in the mice which were infected with the pseudovirions, the luminescent signal intensity for types 16,45 and 58 was 1.779×106p/s, 5.738×105×p/s and 1.829×106p/s, respectively. Conclusion The mouse models for HPV16, 45 and 58 have been successfully established based on pseudovirions, which will be very useful for the research of HPV infection intervention, the evaluation of HPV vaccines and the screening of the prophylactic agents.