Objective To construct and identify the RNAi eukaryotic vector of Skp2 gene and to observe its interfering effect on the growth of SPC-A-1 lung cancer cells.Methods The specific shRNA sequence was designed and synthesized according to the Skp2 cDNA sequence in GenBank.The sequence was cloned into plasmid pGenesil-1.Then recombinant vector was transfected into SPC-A-1 lung cancer cells by Lipofectamine 2000.The expressions of Skp2 mRNA were analyzed by RT-PCR and the levels of Skp2 protein were detected by Western blot.The cell growth suppression was analyzed by MTT assay.Distribution of cell cycle was assessed by flow cytometry.Results The sequence of template and specific siRNA was correct by sequence analysis.Obvious decrease was observed in the levels of Skp2 mRNA and Skp2 protein after Skp2 shRNA transfection(P

Objective To investigate the protective effect of genistein on endotoxin-induced acute lung injury in rats,and explore the underlying mechanisms.Methods Thirty-two male Sprague-Dawley rats were randomly divided into four experimental groups: saline control(S), genistein alone(G),lipopolysaccaride(LPS) alone(L),and genistein pretreatment(G+L).Each treatment group consisted of eight animals.Animals were observed for 6h after LPS challenge,and the wet/dry(W/D) weight ratio of the lung and the protein contents of the bronchoalveolar lavage fluid(BALF) were used as indexes of lung injury.Neutrophil recruitment and activation were evaluated by BALF cellularity and myeloperoxidase(MPO) activity.RT-PCR analysis was performed with lung tissue to assess the gene expression of ICAM-1.The histopathologic changes were also observed using the H&E stains of lung tissue.Results Lung injury parameters,including the wet/dry weight ratio and protein content in BALF,were significantly higher in the L group than in the S group(P

Adolescent ; Calculi ; Humans ; Male ; Nose Diseases ; Turbinates

Objective To study the effects of nitrobenzene on macrophage function and lymphocyte proliferation in mice. Methods ICR mice were divided into groups and treated with nitrobenzene by gavage,once a day,at doses of 2,20 and 200 mg/kg respectively,for 21 consecutive days.The mice were killed after 21 days of treatment and then the effects of nitrobenzene on the organs index,the maerophage function and the lymphocyte proliferation were determined.Results The maerophage function and the lymphocyte proliferation decreased as the increase of the dose of nitrobenzene.Conclusion The results of the present paper show that nitrobenzene may inhibit the immunity function of mice.

Child ; Female ; Humans ; Lymphoma, Extranodal NK-T-Cell

BACKGROUND: With mechanical ventilation widely used in intensive care unit, the ventilator associated pneumonia (VAP) has become a common and serious complication in critically ill patients. Compared with adults, the incidence of VAP and the mortality are higher in children in pediatric intensive care unit (PICU) because of immune deficiency, severe basic diseases, and increased use of artificial airway or mechanical ventilation. Hence it is of significance to study the epidemiology and changes of antibacterial susceptibility in order to reduce the incidence and mortality of VAP in children. METHODS: From January 2008 to June 2010, 2758 children were treated in PICU of Wuhan Children's Hospital. Among them, 171 received mechanical ventilation over 48 hours in PICU, and 46 developed VAP. The distribution and drug-resistance pattern of the pathogenic bacteria isolated from lower respiratory tract aspirations were analyzed. RESULTS: A total of 119 pathogenic microbial strains were isolated. Gram-negative bacilli (G-) were the most (65.55％), followed by fungi (21.01％) and gram-positive cocci (G+, 13.45％). Among them, the most common pathogens were Acinetobacter baummannii, Escherichia coli, Klebsiella pneumoniae, candida albicans and coagulase-negative staphylococci. Antibiotic susceptibility tests indicated that the multiple drug-resistances of G- and G+ to antibiotics were serious. Most of G- was sensitive to ciprofloxacin, amikacin, imipenem, meropenem, cefoperazone-sulbactam and piperacillin-tazobactam. The susceptibility of G+ to vancomycin, teicoplanin and linezolid were 100％. Fungi were almost sensitive to all the antifungal agents. The primary pathogens of VAP were G-, and their multiple drug-resistances were serious. CONCLUSION: In clinical practice we should choose the most sensitive drug for VAP according to pathogenic test.

BACKGROUND: With beta-lactam drugs and immunosuppressants widely used, the infection caused byAcinetobacter baumannii (Ab) has become more and more serious with multidrug resistant Acinetobacter baumannii (MDRAb) emerging and worsening rapidly. Compared with other patients, the incidence and multidrug resistance of MDRAb are higher in children in pediatric intensive care unit (PICU) because of immune deficiency, severe basic diseases, prolonged hospitalization and invasive operations. Hence it is significant to study the epidemiology and changes of antibacterial susceptibility in order to reduce the incidence of MDRAb in children. METHODS: A total 115 patients with MDRAb pneumonia and 45 patients with negative MDRAb (NMDRAb) pneumonia who had been treated from January 2009 to August 2011 were studied retrospectively at the PICU of Wuhan Children's Hospital. Clinical data were analyzed with univariate and multivariate Logistic regression. RESULTS: In 176 clinical strains ofAcinetobacter baumannii isolated, there were 128 strains of MDRAb, accounting for 72.73％. Drug susceptibility tests showed that the resistance rates of β-lactam antibiotics were more than 70％ except for cefoperazone sulbactam. The rates to carbapenems were higher than 90％. They were significantly higher than those of NMDRAb. Amikacin, levofloxacin, ciprofloxacin and minocycline had the lowest drug-resistance rates (<20％). Multivariate Logistic regression revealed that ICU stay, the time of mechanical ventilation, anemia, hypoproteinemia and the use of carbapenems were independent risk factors for MDRAb pneumonia. CONCLUSIONS: MDRAb is an important opportunistic pathogen to pneumonia in PICU, and its drug-resistance is severe. It increases significantly the mortality of patients. It is important to take the effective prevention measures for controlling it.

BACKGROUND: Venous thromboembolism (VTE), including both deep vein thrombosis (DVT) and pulmonary embolism (PE), is a common, lethal disorder that affects hospitalized and non-hospitalized patients. This study aimed to review the progress in the research into VTE. DATA SOURCES: We reviewed the studies about VTE and verified different genetic polymoriphisms of VTE. RESULTS: The pathogenesis of VTE involves hereditary and acquired factors. Many studies indicated that the disorder of coagulation and fibirnolytic system is of utmost importance to this disease. Genetic polymoriphism-related VTE demonstrated significant differences among geographies and ethnicities. CONCLUSION: VTE has many risk factors, but genetic factors play an important role.

Objective To investigate the curative effects of the traditional surgical method of posterior pedicle Schanz screw fixa-tion with partially stripping injured paravertebral muscle in treating thoracolumbar fracture .Methods 52 cases with thoracolumbar fracture underwent posterior pedicle Schanz screw fixation to partially strip injured paravertebral muscle from January ,2010 to Jan-uary ,2012 .The patients were divided into partially stripping injured paravertebral muscle group (32 cases) and traditionally strip-ping injured paravertebral muscle group(20 cases) .The blood loss ,postoperative drainage ,kyphosis postoperative 3 d VAS scores , vertebral height correction rate ,Cobb angle correction ,Oswestry dysfunction indexes were copmared .Results Comparing with tra-ditionally stripping injured paravertebral muscle group ,the partially stripping injured paravertebral muscle group has better effect in transperative blood loss ,postoperative drainage and postoperative 3 d VAS scores ,the differences were statistically significant .But there was no statistical significance between the two groups in vertebral height correction rate ,kyphosis Cobb angle correction ,Os-westry dysfunction indexes .Conclusion Partially strip injured paravertebral muscle has less painful than traditional surgery does , being safe and effective with small invasion and easy to recover .

Objective:To establish the RPB5-mediating protein (RMP)-silenced stable cell lines and study the inhibitory effects of small interfering RNA (siRNA) targeting RMP gene on the proliferation and migration of human hepatoma SMMC-7721 cells. Methods:Three RMPi siRNAs were designed and synthesized in vitro and transfected into SMMC-7721 cells. The inhibitory effect of siRNA on RMP gene expression was measured by RT-PCR to select the best siRNA. The expression vector pGPU6-Neo-RMP-484 was transfected into SMMC-7721 cells by the lipofectamine and the cells stably expressing the siRNA were selected by G418. RT-PCR was used to detect the interference efficacy against RMP gene. Cell proliferation and adhesion were measured by MTT assay. Wound healing test was used to observe the migration ability of cells. Results:The SMMC-7721 cell lines with down-regulated RMP expression were established by using RNA interference technology. Compared with the negative control cells, expression of RMP mRNA was down-regulated by(83.67±2.56)% .The proliferation of stable-transfected cells was inhibited by(74.33±0.58)% . The adhesion capability of stable-transfected cells was enhanced but the migration capacity was decreased compared with the negative control cells. Conclusion:The pGPU6-Neo-RMP-484 cell lines with stable transfection of RMP siRNA recombinant vector are successfully screened,which can be used as a cellular model for studying the molecular mechanism of RMP. Down-regulation of RMP gene expression can effectively inhibit the proliferation, enhance the adhesion, and decrease the migration of SMMC-7721 cells.