OBJECTIVE For the reasonable use of antibacterial drugs, drug resisitance diversity of Pseudomonas aeruginosa was studied in our hospital. METHODS Antibacterial activity to all P. aeruginosa collected in four years from 2004 to 2007 were determined by K-B test, and data were analyzed by WHONET software. RESULTS The resistance rate to ampicllin, cefazolin and cefoteton were 100.0%, but low to ceftrazidime, cefoperazone and cefoperazone/sulbactam(3.2% in 2007). The resistance rate to levofloxacin increased year by year, while that to ciprofloxacin descreased from 15.6% to 9.6%.The resistance rate to gentamicin and amikacin descreased from 20.0% to 16.4%,and from 11.1% to 4.8%, respectively. The resistance rate to imipenem and meropenem accounted for about 10.0%, which was as same as ceftazidine. CONCLUSIONS The resistance of P. aeruginosa in our hospital is stable. Clinician should choose right antibacterial drugs on the basis of the test for antibacterial sensitivity and the pharmacological characteristis to improve curative effect and decrease the resistance of bacteria.

Abstract] Objective To establish an assay for the detection of porcine parvovirus ( PPV) and to verify its application for monitoring cells used for production.Methods A pair of primers and one probe were designed according to the conserved sequence encoding non-structural protein 1 (NS1).Based on the designed primers, a real-time fluorescent quantitative PCR assay for the detection of PPV was developed. Several parameters including the linearity, precision, minimum detection limit and anti-interference of the established assay were evaluated.A stock of PPV strains was prepared by infecting swine testicle ( ST) cells with PPV strains.An assay for the detection of PPV infection was developed by using ST cells as sensitive cells.A combined ST cell infection-PCR test was developed by combining the ST cell infection assay with the real-time fluorescent quantitative PCR assay.The sensitivity of ST cell infection-PCR test was analyzed.The cell samples used for production of biological products were detected by using the established assay.Results The real-time fluorescent quantitative PCR assay was specific for the detection of PPV without cross-reaction to other species of parvovirus virus, SV40 virus and other porcine viruses.The linear range of the assay was 1×109-1×104 copies/μl with a R2 value more than 0.98.The sensitivity of the real-time quantitative PCR assay was 1×104 copies/μl.Both of the intra-and inter-coefficient of variation (CV) were less than 5%in Ct values.The intra-and inter-CV in copies of detection were 5%-15% and 30%-40% respectively.The minimum detection limit of the real-time fluorescent quantitative PCR assay was 1CCID50/ml.The PPV strains were detected in cell samples with no interference.The sensitivity of ST cells infection-PCR test was 0.01CCID50/ml.All of the 22 cell samples were negative for PPV by using the real-time fluorescent quanti-tative PCR assay.Conclusion The real-time fluorescent quantitative PCR and the ST cell infection-PCR test for the detection of PPV in cells were established successfully.The application of the two assays was conducive to further enhance the safety of using cells for production and therapy.

Objective To evaluate the clinical significance of the changes of microalbunminuria(MAU) levels in mechanical ventilated patients with severe pneumonia. Methods According to the ratio between the microalbunminuria and the urine creatinine (MAU/CR) (ACR), setting 25mg/mmol as the threshold, 78 mechanical ventilated patients with severe pneumonia were divided into two groups :ACR increasing group and ACR Non-increasing group,then the clinical significance of changes of MAU levels in 72 hours on prognosis of these patients was observed. Results MAU increased in 64 cases(82. 1%) ,of which 46 cases in ACR increasing group and 18 cases in ACR non-increasing group. There showed statistically significant differences on APACHE Ⅱ score, CPIS score,PCT、the success of getting out of mechanical ventilation and the mortality between two groups, (t = 3.50、2. 19 、x~2 = 3. 95、6. 70、5.38 ,P = 0.01,0.03,0.04,0.01,0.02, all P ＜ 0.05). Conclusion Changes of MAU levels have the clinical significance on prognosis of the mechanical ventilated patients with severe pneumonia.

Objective To evaluate five detection methods for the laboratory diagnosis of Clostridium difficile infection in the hospitals of USA, and explore a sensitive, specific, accuracy and rapid regimen for the early diagnosis of Clostridium difficile infection. Methods A total of 174 stool specimens submitted to the clinical microbiology laboratory for Clostridium difficile testing were separately tested by five methods including toxigenic culture (TGC), Premier Toxin A&B EIA( A/B-EIA), C. Diff Quick Chek Complete( DEIA), BD G eneOhm Cdiff assay(BD-PCR) and Laboratory-developed PCR(LD-PCR). The gold standard of TGC was used as a reference criterion, and the sensitivity, specificity, positive predictive value ( PPV )and negative predictive value (NPV) of A/B-EIA, D-EIA, BD-PCR and LD-PCR assays were determined. Results Among the 174 specimens studied, 24 were defined as true positives for Clostridium difficile infection by TGC assay, giving a positive rate of 13.8% (24/174). In comparison to the standard,the sensitivity, specificity, PPV and NPV were 62.5%, 99.3%, 93.8% and 94.3% for A/B-EIA;66.7%, 98.7%, 88.9% and 94.9% for D-EIA; 83.3%, 98.7%, 90.9% and 97.4% for BD-PCR;79.2%, 93.3%, 65.5% and 96.6% for LD-PCR. Among all tested specimens, 34 were positive by atleast one of five methods, and of which 15 were concordant by all five methods. The D-EIA results were divided into three groups depending on results of GDH and (or) toxins A/B: 18 were positive for both GDH and toxins A/B, 23 were positive for only GDH, and 133 were negative for both GDH and toxins A/B. Of 18 positive specimens by D-EIA assay, all were concordant with results of BD-PCR assay and 16 were agreement with results of TGC assay. Twenty-two of 24 positive specimens by TGC assay were included in 41 specimens that were positive for GDH. Among eight false negative specimens by D-EIA assay, four were differentiated as positive results by BD-PCR. According to the present study, the sensitivity, specificity,PPV and NPV of a two-step detection algorithm in combination with D-EIA and BD-PCR assays were 83.3%, 98.7%, 90.9% and 97.4%, respectively. Conclusions From the point of technological evaluation, BD-PCR is preferable. A two-step detection algorithm combining D-EIA with BD-PCR is proposed for the laboratory diagnosis of Clostridium difficile infection. This algorithm has demonstrated an excellent sensitivity and specificity, as well as decreased test turnaround time and test cost.

Objective:To provide the basic data for the further revision of mycoplasma test method described in Chinese Pharma-copoeia and some references for the operation standardization of drug mycoplasma test. Methods:Two incubation conditions,namely aerobic conditions and microaerophilic conditions,were compared with respect to the growth status of mycoplasma in liquid and solid media. Results:The growth of mycoplasma was obvious difference between the two incubation conditions,and the microaerophilic con-ditions were better than the aerobic conditions. Conclusion:The microaerophilic conditions can be used in the incubation of mycoplas-ma test,which should be defined and standardized in the future Chinese Pharmacopoeia.

Objective:To investigate the growth-promoting properties and applicability of the mycoplasma test culture media pre-scribed in Chinese pharmacopoeia, and provide reference for the standardization of the drug mycoplasma test method. Methods: Re-spectively using four quantitative detection methods including sensitivity, degree of color change, colony counts and colony diameter, and mycoplasma media widely used in the world as the reference media, the growth-promoting properties of 4 batches of mycoplasma broth and 4 batches of arginine mycoplasma broth from four domestic manufacturers were investigated. Results:The results of sensitivity assay and absorbance detection showed that all the media inoculated with below 100 CFU test microorganisms exhibited visible color change. Furthermore, the results of color change degree and colony diameter showed that there were significant differences among the media products from different manufacturers(P<0. 01). Conclusion:Mycoplasma broth and arginine mycoplasma broth both can sup-port the growth of below 100 CFU test microorganisms. Due to the difference in the growth-promoting properties among the media prod-ucts from different manufacturers, the drug mycoplasma test workers should use more sensitive methods to examine the applicability of the media.

Objective To study the in vivo expression and biodistribution of Ad5-Fluc (Adenovirus carrying firefly luciferase genes) in mice.Methods The recombinant Ad5-Fluc virus was constructed and infected to BALB/c or nude mice through three different routes.The protein expression level,tissue distribution and the characteristics of infection were analyzed by in vivo bioluminescence imaging technology.Results Compared to other two routes,the BALB/c mice infected through muscular route had the longest expression cycle (over 60 days) and the highest expression level,while the virus was transferred into the liver and spleen after infection.The nude mice had a significantly extended expression cycle than BALB/c mice.Moreover,the characteristic of liver tropism was eliminated after Ad5 F35 infection in mice,while maintained similar expression efficiency.Conclusion Due to the highest expression efficiency,the muscular route would be the optimal route for Ad5 vector based vaccination.In addition,Ad5F35 virus could become an ideal alternative vaccine vector for eliminating the liver tropism.

Objective To investigate the diagnostic potential of magnifying narrow-band imaging endoscopy (NBI-ME) for different intrapapillary capillary loop (IPCL) for the diagnosis of esophageal lesion.Methods Patients with abnormal esophageal mucosa found by white light gastroscopy in digestive endoscopy center,Chinese PLA General Hospital during the period of November 2009 to November 2010 were enrolled in this study.IPCL was observed and divided into different types by NBI-ME.Histopathology of biopsy or endoscopic submucosal dissection (ESD) specimens was evaluated and used as the gold standard to evaluate the diagnostic value of NBI-ME for IPCL.Results A total of 146 lesions from 145 subjects with esophageal mucosa abnormal were collected. Among them, 88 were pathology-proven inflammation,5 were pathology-proven esophageal cancers,20 were pathology-proven low intraepithelial neoplasia (LIN) and 33 were pathology-proven high intraepithelial neoplasia (HIN) detected with NBI-ME.By a per-lesion analysis,the accuracy of inflammation and cancer were 100％ (88/88) and 7/7.For the sensitivity,specificity,accuracy,positive predictive value,negative predictive value,positive likelihood ratio,negative likelihood ratio of LIN and HIN were 7/10,69.8％ ( 30/43 ),69.8％ ( 37/53 ),35.0％ (7/20),90.9％ (30/33),12.5％ (70/559),2.3％ (30/1290) and 87.1％ (27/31),72.7％ ( 16/22),81.1％ ( 43/53 ),81.8％ ( 27/33 ),80.0％ ( 16/20 ),634.1％ ( 837/132 ) and 35.2％ ( 124/352 ),respectively.Conclusions NBI-ME can classify the different esophageal IPCL.Higher diagnostic accuracy of IPCL indicates the feasibility of NBI-ME for the efficacious diagnosis of esophageal inflammation and cancer.There is the higher diagnostic accuracy of HIN than LIN.

Type 2 diabetes macrovascular disease is the main cause of death in type 2 diabetes mellitus. In recent years, the modern medical research and treatment of type 2 diabetes macrovascular disease has made some progress, but the international clinical trials suggest that the current treatment can not effectively reduce the incidence of this disease. Many clinical practices show that the effect of traditional Chinese medicine on this disease is exact, so that the clinical workers on the treatment of type 2 diabetes mellitus macrovascular disease of the status quo, now from the etiology and pathogenesis,clinical research, experimental research on the literature published in recent years, to provide reference for clinical treatment.

Objective To investigate the effect of triptolide (TP) on proliferation and apoptosis of cyst-lining epithelial cells with autosomal dominant polycystic kidney disease ( ADPKD). Methods Primary cultured cyst-lining epithelial cells were treated with TP at different concentrations for 12 h,24 h,48 h and 72 h, respectively.The proliferation activity of the cells was evaluated by Brdu assay. The cell cycle distribution was determined by flow cytometry. The apoptotic and apoptotic ratio were determined by FITC-AnnexinV binding/ PI. The morphological changes of cyst-lining epithelial cells were observed under transmission electron microscope. Results TP significantly inhibited the proliferation of cyst-lining epithelial cells and induced apoptosis in a dose- (10-40 ng?mL-1 )and time-dependent(12-48 h) manner. Typical ultrastructural changes of apoptotic cells were observed under electron microscope. Conclusion TP significantly inhibited the proliferation of cyst-lining epithelial cells and induced the apoptosis of cyst-lining epithelial cells, thus inhibited cyst forming and delayed cyst developing. The mechanism may involve several targets and pathways.