Objective To investigate the distribution and resistant pattern of Enterobacter cloacae to antimicrobial agents for reasonable use of antibiotics .Methods E .cloacae strains were isolated from patients from January 2009 to December 2013 .The strains were identified by VITEK‐2 Compact System of BioMerieux and tested for antimicrobial susceptibility by Kirby‐Bauer disk diffusion method .The results were analyzed .Results A total of 397 nonduplicate E .cloacae strains were isolated during 5 years ,accounting for 5 % of the total gram‐negative isolates . The strains were mainly isolated from sputum (48 .9% ) , followed by secretions (30 .5% )and urine (7 .3% ) .The percentage of E .cloacae strains resistant to all the antibiotics tested was on decline except carbapenems ,cefoxitin and amoxicillin‐clavulanic acid .Carbapenems were relatively more active against E .cloacae strains .The E .cloacae strains showed higher resistance rate toβ‐lactams .Conclusions It is necessary to strengthen the monitoring of clinical isolates to guide rational use of antimicrobial agents .

Objective To investigate the effect of thermochemotherapy with 5 fluorocytosine(5 FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue specific cytosine deaminase (CD) in vitro.Methods Recombinant retroviral vector G1CEACDNa, in which CD gene was controlled under the CEA promoter, was introduced through liposome technique to the human colorectal carcinoma cell line SW480, and then the cells were selectively cultured in G418. The proliferated colonies were treated with the combined therapy of 5 FC and hyperthermia at a 43℃ over 30 min for 3 times. RT PCR was performed to analyze the expression of CD gene in target cells after thermochemotherapy. Results The CD gene expressed steadily in the target cells after 3 times of hyperthermic treatment. After the transfection of CD gene, SW480 CEACD cells were more sensitive to 5 FC than their parental cells (P

Objective To study the mechanism of SW480 cell line death caused by transfection of retroviral vector-G1CEACDNa.Methods Plasmid G1CEACDNa was transferred into the SW480 cell line using liposomes method. RT-PCR was performed to examin the expression of CD gene indrectly.The cell growth curve was measured by means of cell counting. When the CD+SW480 cells were exposed to 5-FC (1mmol/L), the growth inhibition rate and the "bystander effect" were detected by MTT method.The ultrastructure was observed by electron microscope.Apoptosis was verified by flow cytometer .Results The product of RT-PCR showed a band at 1.5kb on the photo of electrophoresis. The growth of CD+ SW480 cells was inhibited 24h after administrating 5-FC,and the inhibition rates at 72h,96h,120h were 30.0%,50.0% and 80.0%,respectively.Apoptosis cells in different phases and apoptotic bodies in the field of electron microscope were observed. Meanwhile ,a few cells showed necrosis.Flow cytometer verified that a few cells appearred apoptosis 48h after exposed to 5-FC (1mmol/L), the apoptosis rate and the necrosis rate at 72h,96h were 20.2%,30.7% and 19.6%,21.1% respectively.Conclusions The death mechanism of SW480 cells transfected with G1CEACDNa followed by 5-FC treatment includes both necrosis and apoptosis.Apoptosis is possibly the bystander effect.

Objective To use the liquid protein combined with MALDI-TOF-MS for screening the serum differential peptides markers in lung adenocarcinoma patients and to establish the lung adenocarcinoma diag-nosed prediction model for founding the potential markers for the diagnosis of lung adenocarcinoma.Methods 37 patients with lung adenocarcinoma and 33 healthy subjects and benign lung disease which were made up in control group were collected,in the two groups the age and the sex were matched.The two groups were ran-domly divided into training group(30 cases of lung adenocarcinoma,26 cases of control)and test group(7 ca-ses of lung adenocarcinoma,7 cases of control)according to 3:1.T he differential diagnosis of lung adenocarci-noma and control group was performed by liquid chip-time-of-flight mass spectrometry and software ClinPro-Tools 3.0 to establish a prediction model of lung adenocarcinoma.The diagnostic model was validated by using serum samples from the test group to assess the diagnostic efficacy of the model.Results Nine peptide peaks with significant differences(P<0.05)were obtained by ClinProTools 3.0 software analysis.The up-regulated peaks in lung adenocarcinoma(m/z)were 8 976.5,4 469.05,4 966.78,8 925.5,4 531.05,and the down-reg-ulated m/z were 3 304.44,8 594.76,3 266.82,3 195.52.According to the genetic algorithm(GA),the lung ad-enocarcinoma diagnosis and prediction model was established.The overall recognition ability of the model was 94.49%.The model was evaluated by the test group.The results showed that the sensitivity of the model was 100.0% and the specificity was 85.7%.Conclusion Among lung adenocarcinoma patients,serum benign lung disease and healthy,there are differences in the serum peptide.T he use of differential peptide peaks to estab-lish lung adenocarcinoma diagnostic prediction model for the early diagnosis of lung adenocarcinoma provides a new method.

Objective To observe the effects of ice-water swimming on pathological changes in model gouty rats,and investigate the relevant regulatory mechanism of the purinergic P2X7R receptor.Methods Male Sprague Dawley rats were divided into normal(NORM)and experimental groups including gouty control(GC),ice-water swimming(IWS),and Brilliant Blue G(BBG,a P2X7R inhibitor)groups.Rats in the experimental groups were modeled to simulate hyperuricemia and gouty arthritis by inhibiting uric acid metabolism combined with the Coderre method.Rats in the ice-water swimming group were treated with 5 min of endurance swimming in an ice-water mixture at a depth of about 0.5 m for 0 h and 12 h after modeling by the Coderre method,while rats in the BBG group were injected intraperitoneally with BBG solution once after modeling.Ankle swelling index was calculated using a formula.Serum uric acid levels were detected by uricase assay,and serum levels of the inflammatory factors interleukin(IL)-1β,1L-6,and tumor necrosis factor(TNF)-αwere detected by enzyme-linked immunosorbent assay.The pathological status of the ankle joints was examined by hematoxylin and eosin staining.P2X7R and NLRP3 protein expression levels in synovial tissue were detected by Western blot and immunohistochemistry,respectively.Results Serum uric acid levels and the ankle joint swelling index were significantly higher in the experimental groups compared with the normal group(P＜0.05 or P＜0.01),and the synovial tissues showed different degrees of inflammatory infiltration.The ankle swelling index was significantly higher in the ice-water swimming group compared with the gouty control group at 12 h(P＜0.05).Serum IL-1β,IL-6,and TNF-α levels(P＜0.01)and P2X7R and NLRP3 protein levels in synovial tissues were all significantly elevated(P＜0.05).Histopathology showed that the cartilage surface was broken and the synovial tissue showed severe hyperplasia and erosion,accompanied by numerous inflammatory cell aggregates.There were no significant changes in P2X7R or NLRP3 protein expression or pathology in synovial tissues in the BBG group compared with the gouty control group(P＞0.05),but serum IL-1β,IL-6,and TNF-α levels were all significantly suppressed(P＜0.01).Conclusions Cold stimulation and strenuous exercise simulated by ice-water swimming may exacerbate pathological damage in gouty arthritis via a mechanism related to high P2X7R expression in the joints.

Objective: To establish a diagnostic prediction model for esophageal squamous cell carcinoma (ESCC) and search the potential biomarkers of ESCC. Methods: Serum samples from 59 patients with ESCC and 57 healthy controls were collected, and randomly divided into the training group (44 patients and 42 healthy controls) and validation group (15 patients and 15 healthy controls). Serum proteins/peptides were extracted and purified with weak cation-exchange chromatography Magnetic Beads (WCX-MB), and detected by the matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). Then the differentially expressed proteins/peptides were screened out, and a diagnostic prediction model for ESCC was established and preliminarily validated. Results: The ClinProTools software identified 31 differential peptide peaks (P＜0.05), among which 18 peaks had significant difference (P＜0.01). Compared with healthy controls, 8 peaks were up-regulated in ESCC patients, while 10 peaks were down-regulated. Among them, the areas under the receiver operating characteristics (ROC) curve (AUC ROC ) of m/z 2 660.84 and m/z 5 336.49 peaks were 0.95 and 0.91, respectively, and their expressions were up-regulated in ESCC patients. The validation results showed that the accuracy, sensitivity and specificity of the diagnostic prediction model established by the genetic algorithm (GA) were 93.10%, 92.90% and 93.30%, respectively. Conclusion The established diagnostic prediction model may be used for the auxiliary diagnosis of ESCC. Two peptide peaks of m/z 2 660.84 and m/z 5 336.49 may be the potential biomarkers of ESCC.