Objective To investigate the mutations of quinolone resistance determinational region ( QRDR) in fluoroquinolon-resistant Pseudomonas aeruginosa strains isolated from patients with nosocomial pneumonia. Methods Eight-four Pseudomonas aeruginosa strains isolated from patients with nosocomial pneumonia in Xinhua Hospital during January 2006 to December 2007, from whom fluoroquinolon-resistant resisitant ( case) and fluoroquinolon-susceptible ( control ) Pseudomona aeruginosa were identified. The mutation of QRDR was tested by restriction fragment length polymorphism ( RFLP) and gene sequencing.The relationship between QRDR mutations and clinical prescription was analyzed. Results Mutation in QRDR was found in 42 isolates among the 50 fluoroquinlon-resisitant isolates( 84. 0% ) , while no mutation was found in fluoroquinlon-susceptible isolates. The mutation in GyrB Ser464 was found in 34 isolates ( 68. 0% ) . There was statistical difference in the usage of β-lactams between the GyrB-Ser464-mutated group and the non-GyrB-Ser464-mutated group( OR = 11. 3, P = 0. 003 and OR = 3. 5, P = 0. 023) , also in the time of fluoroquinolon usage before isolated ( P = 0. 038) . Conclusions The mutation of QRDR is contributing to fluoroquindor-resisitance of Pseudomona aeruginosa, most of which lies in GyrB Ser464.Abuse of β-lactams and fluoroquinolon may be the risk factors of mutation in GyrB Ser464.
Objective To investigate the effects of down-regulating of Rfng gene ( 1, 3-Nacetylglucosaminyltransferases) in lung CD4 + T cells of asthmatic rat model by small interfering RNA ( siRNA) and explore the role of Rfng in pathogenesis of asthma. Methods An asthmatic rat model was established by OVA sensitization and challenge. Total T cells were isolated from lung tissue of asthmatic rats, and CD4 + T lymphocytes were purified using magnetic beads. CD4 + T lymphocytes were transfected by siRNA targeting Rfng gene. The mRNA and protein expressions of Rfng were detected by quantitative PCR and Western blot. Quantitative PCR was performed to determine the mRNA levels of Th1 /Th2 cytokines and related genes including IL-12, IFN-γ, IL-4, IL-5, T-bet, and GATA3. ELISA was performed to determine the concentrations of IL-12, IFN-γ, IL-4, and IL-5 in supernatant. Results The mRNA and protein expression of Rfng in RNAi group decreased significantly. IL-12, IFN-γ, T-bet increased and while IL-4, IL-5, and GATA3 decreased significantly. The concentrations of IL-12 and IFN-γ in the supernatant increased significantly, while IL-4 and IL-5 decreased significantly. Conclusions Down regulation of Rfng affects T cell differentiation. It is presumed that Fringe contribute to the pathogenesis of asthma.
Objective Bone marrow mesenchymal stem cells （MSCs） have been suggested to play an important role in the treatment of a variety of pulmonary diseases. The present study was aimed at evaluating the therapeutical effect of MSCs transplantation on emphysematous rats，and explore its influence in local and systemic inflammation. Methods Emphysema rat model was established by cigarette smoking. MSCs were transfected with lentivirus vector carrying green fluorecent protein （GFP） and the transfected MSCs in lung of smoke rats were detected by imaging system for small animals. Thirty-six SD rats were randomly divided into a control group，an emphysema group，and a MSCs transplantation group. The total and differential cell counts in bronchoalveolar lavage fluid （BALF） were measured. TNF-α and IL-1β levels in BALF and serum were measured by ELISA. Malonaldehyde （MDA） level in lung tissue was detected by chromatometry. Emphysema changes were evaluated by mean linear intercept （MLI） of lung under light microscope by HE staining. Results The transfected MSC in different lung lobes were found to be alive at four weeks after intrapulmonary delivery. Compared with the emphysema group，the total cell count in BALF，TNF-α and IL-1β levels in BALF and serum，MDA level in lung tissue and MLI were significantly reduced intheMSCs transplantation group（Plt;0.01）. Conclusions Transplantation of MSCs can mediate down-regulation of TNF-α and IL-1β in BALF and serum，attenuate inflammation，oxidative stress and emphysema change of lung，suggesting that MSCs have significant therapeutic effects on emphysema.
Objective To evaluate the subjective outcomes of sleepiness behavior and mood status applying continuous positive airway pressure（CPAP） in adults of elderly and middle-aged with obstructive sleep apnea syndrome（OSAS）. Methods Nine randomized controlled trails comparing nocturnal CPAP with inactive control appliances in adults with OSAS with the use of computerized search in related medical databases（MEDLINE，EMBASE，CBMdisk，etc） were included.The quality of literature was reviewed，and all data were extracted by two reviewers independently.Meta analysis was conducted used RevMan 4.2 software.Results 9 RCT involving 665 patients of elderly and middle-aged met the inclusion criteria.Meta analysis indicated that the score of Epworth sleepiness scale（ESS） and general health questionnaire-28（GHQ-28） declined significantly after CPAP treatment on effectiveness with WMD（random） －2.94，95 %CI －4.68 to －1.20，or WMD（fixed） －2.26，95 %CI －3.79 to －0.72，Plt;0.01.Nevertheless，hospital anxiety and depression scale（HADS） was not significantly different between CPAP and control with WMD（random） －0.89，95%CI －1.98 to 0.20，Pgt;0.05.Conclusion Current clinical evidence suggested that CPAP was effective in improving day-time subjective outcomes of sleepiness behavior and general mental health status in OSAS patients of elderly and middle-aged，although evidence of improving emotion disorder of anxiety and depression was not confirmed.
Objective To analyze the current drug resistance and risk factors of hospital acquired pneumonia( HAP) due to extended spectrumβ-lactamase ( ESBLs) producing Escherichia coli and Klebsiella pneumoniae, and to estimate the prevalence trend of ESBLs producing strains. Methods FromApril 2007 to January 2008, 140 patients of Xinhua Hospital with HAP due to E. coli and K. pnermoniae were enrolled.Among them, 88 patients were with ESBLs producing strains and 52 patients were with non-ESBLs producing strains. Risk factors were analyzed by comparing between these patients. The rate of drug resistance was determined by antibiotic sensitive test. Fifty-three ESBLs producing strains were genotyped by random amplified polymorphic DNA ( RAPD) . Results The rate of drug resistance of ESBLs producing strains washigher than that of non-ESBLs producing strains. ICU stay, use of third- and forth-generation cehpalosporin were found to be the independent risk factors by multivariate analysis with logistic regression. By RAPD, 37 ESBLs producing E. coli strains were divided into 27 types and 16 ESBLs producing K. pneumoniae strains were divided into 13 types. Conclusions ICU stay, use of third-generation and forth-generation cehpalosporin remain as major risk factors in the HAP due to ESBLs producing E. coli and K. pneumoniae.RAPD is an economic, quick and credible method for epidemic analysis
Objective To investigate the expression of the histone deacetylases 1( HDAC1) and the level of whole histone acetylation and methylation in lung T cells of asthmatic rats, and investigate their role in the pathogenesis of asthma.Methods Sixteen wistar rats were randomly divided into a control group and an asthma group( n =8 in each group) . The rats was sensitized with ovalbumin( OVA) and challenged with aerosol OVA to establish asthma model. The asthmatic ratmodel was confirmed by measurement of pulmonary function, histochemical staining, HE staining, and the levels of interleukin-4 ( IL-4 ) , interferon-gamma ( IFN-γ) and immunoglobulin E( IgE) in serum and bronchoalveolar lavage fluid ( BALF) . T cells were isolated fromrat lungs and the purity was identified. The expression of the HDAC1, the level of whole histone H3 and H4 acetylation, and whole H3K9 dimethylation were analyzed by Western blot in lung T cells. Results Compared with the control group, the protein expression of HDAC1 was significantly lower( 0. 465±0. 087 vs 0. 790 ±0. 076, P lt;0. 05) in lung T cells of the asthma group. No significant differences werefound in regard to the level of whole histone H3 and H4 acetylation and whole H3K9 dimethylation betweenthe two groups. Conclusions HDAC1 in lung T cells may be involved in the pathogenesis of asthma.Histone modification by HDAC1 may be a specific eventwith gene transcription which may not be associated with asthma.
Objective To examine the expression of promoter CpG island methylation of Notch1 gene and explore the variable sites for DNA methylation in lung CD4 + T cells of asthmatic rat models.Methods An ovalbumin ( OVA) sensitized- challenged asthmatic rat model was established. Total T cells were isolated and CD4 + T lymphocytes were purified using magnetic beads. Twenty Wistar rats were randomly divided into a control group and an asthma group ( n = 10 in each group) . CD4 + T cells were isolated by immunomagnetic beads and identified by flow cytometry ( FCM) . Realtime PCR was employed to examine the mRNA expression of Notch1 gene in lung CD4 + T cells and the methylation level of Notch1 gene was examined by methylation-specific PCR. Results The mRNA expression of Notch1 in lung CD4 + T cells of the asthma group was 2. 254 ±0. 403 times as much as that of the control group. The total methylation level of asthma group was lower than that of the control group ( 0. 150 ±0. 108 vs. 0. 300 ±0. 667, P lt;0. 01) . Conclusion Promoter demethylation is one of the major mechanisms of Notch1 gene up-regulation in pathogenesis of asthma.
Objective To investigate the role of red cell distribution width ( RDW) in coronary artery diseases patients complicated with obstructive sleep apnea-hypopnea syndrome ( OSAHS) . Methods 134 coronary artery diseases patients who had at least one-vessel disease confirmed by coronary angiography were investigated by polysomnography for OSAHS. The patients were classified according to theapneahypopnea index(AHI) . The level of RDW, triglyceride, cholesterol, high density lipoprotein, low density lipoprotein, hemoglobin, white blood cells and hematocritwere measured. The receiver operating characteristic curve was drawn to predict the moderate-severe OSAHS in coronary artery diseases patients according to RDW value. Results When 134 coronary artery diseases patients were classified into a control group and an OSAHS group according to the AHI, the level of RDW in two groups was not significantly different [ ( 13.44 ±1.30) % vs. ( 13.12 ±0.92) % , P gt; 0.05] . When 134 coronary artery diseases patients were classified into a control and mild OSAHS group and a moderate-severe OSAHS group according to the AHI, the level of RDW in two groups was significantly different [ ( 13.07 ±0.94) vs. ( 14.02 ±1.41) % , P lt; 0.05] . And no difference was found in hemoglobin, triglyceride, cholesterol, high density lipoprotein, low density lipoprotein, platelet, and hematocrit between two groups. The ROC curve analysis revealed that the area under ROC curve was 0.748 ( 0.523-0.972) , and the best cut-off for moderate-severe OSAHS was 13.95% with sensitivity of 71.43% and specificity of 82.98% . Conclusion RDW may be a useful and simple tool to predict moderate-severe OSAHS in coronary artery diseases patients.
Objective To observe the immune responses of T helper cells 17 ( Th17) to respiratory syncytial virus ( RSV) infection induced lung inflammation in mice, and explore its roles on the host immune responses to RSV.Methods Female BALB/ c mice aged 3 to 5 weeks were randomly divided into a RSV group ( n=18) and a control group ( n = 12) . The mice were intranasally administrated by a 107.5 50% tissue culture infective dose ( TCID50) of RSV in 0.1 mL of culture medium. Sterile medium ( 0.1 mL/ mouse) was used as control. After infected on 1st , 4th, 8th day, the mice were sacrificed, and specimens from the lungs and lymph nodes were collected. The lung sections were stained by hematoxylin-eosin to observe the changes of lung inflammation after RSV infection. IL-17A, IL-17F and IL-23p19 mRNA expressions in the lung tissue were determined by real-time PCR. The frequencies of Th17 subsets in hilar lymph node were analyzed by flow cytometry. Results On 4th day after RSV infection, a typical lung interstitial inflammation was observed. However, this inflammation was alleviated on 8th day after RSV infection. The viral load in the lung tissue on 4th day after RSV infection were 9.208 ±0.548, which was the highest among all RSV subgroups ( P lt;0.001) . IL-23p19 and IL-17A cytokine expressions in the lung tissue were significantly increased on 4th day and 8th day after RSV infection compared with control groups ( P lt;0.01) , and the peak was on 4th day. However, IL-17F mRNA expression in the lung tissue on different day after RSV infection had no significant difference compared with the control group ( P gt;0.05) . The frequencies of Th17 subsets in hilar lymph node on 4th day and 8th day after RSV infection were ( 0.37 ±0.043) % and ( 0.853 ±0.048) % respectively, which were higher than those in control groups ( P lt;0.05) . The frequencies of Th17 on 8th day after RSV infection were significantly higher than that on 4th day after RSV infection ( P lt; 0.01) . Conclusions The expression of IL-17A in the lung tissue is increased and the level of Th17 cells in hilar lymph nodes is also elevated in the lung infected by RSV, which indicates that Th17 cells might be involved in host antiviral immune.
Objective To investigate the clinicopathologic features, diagnosis, treatment and prognosis of the primary lymphoepithelioma-like carcinoma (PLELC) of the lung. Methods Clinical and pathological data of one patient with PLELC of the lung were reviewed. PLELC of the lung was analyzed through review of literatures. Results A mass in the upper lobe of the left lung and atelectasis were detected by CT in a 36 year old nonsmoker male. He presented with cough, expectoration, fever and shortness of breath for one month. Neoplasm in left main bronchus was observed through bronchoscopy. Histological study showed cohesive clusters of comprising syncytial-appearing cells with vesicular nuclei and tumor infiltrating lymphocytes. Immunohistochemistry showed positive expression of Epstein-Barr virus (EBV) encoded small nuclear RNA. Nasal CT scan was negative for suspicious nasopharyngeal mucosal lesion or tumor. Then he was diagnosed as PLELC. The patient received docetaxel+nedaplatin regimens for 6 cycles with stable disease, but one month later after the end of chemotherapy, he was readmitted to hospital for dyspnea. CT scans showed progression of the lesion with massive pericardial effusion and pleural effusion. The patient could not tolerate chemotherapy, then received supportive treatment and died soon. Totally 18 Chinese articles and 25 foreign articles were screened out. Literature review suggested that patients with PLELC of the lung usually occurred in young nonsmokers and without gender difference. PLELC of the lung was closely associated with EBV infection. Its clinical manifestations were not specific, so that histopathological and immunohistochemical analyses were the main method of diagnosis. PLELC of the lung may respond to chemotherapy and radiotherapy. Surgery-based multimodality treatment was recommended. Patients with early or resectable disease had better prognosis, but patients with unresectable disease had poor prognosis. Conclusions PLELC of the lung may be closely associated with EBV infection. Histopathological and immunohistochemical analysis is the main method of diagnosis. Surgery-based multimodality treatment should be recommended because of its high response to chemotherapy and radiotherapy. Patients with early or resectable disease have better prognosis.