Objective To amplify the 16S RNA fragments of 7 clinically isolated strains of Brucella spp. by PCR-RFLP technique, so as to provide experimental basis for the studies on diagnostics, genetics and epidemiology of Brucella spp. Methods According to the gene sequence of ATCC 25840 standard strain in GenBank, special primers for the 16S RNA conservative area in the Brucella spp. were designed. DNA extraction and PCR amplification of the 16S RNA fragments were performed with the 7 isolated strains. PCR products were then sequenced and RFLP analysis was conducted with appropriate restricted enzymes to study the homology and the mutation sites in those strains. Meanwhile, the clinical data of infected patients were retrospectively analyzed to evaluate the relationship between the clinical features and genotypes of Brucella infection. Results The amplified target fragments were about 1500bp in length and consistent with what was expected. The sequencing and homology analysis showed a 98.88% homology and 11 mutation sites among the 7 isolated strains. Four genotypes were identified by RFLP. Retrospective analysis of the clinical data indicated that no obvious relationship existed between the genotypes and the clinical features. Conclusions Amplifying 16S RNA fragments by PCR technique is a feasible method to make an early diagnosis of Brucella infection. The 7 clinically isolated strains are different in genotypes and 16S RNA fragment is a highly conservative fragment in bacterial genome with some mutations. The research provides evidence for the genetics and epidemiology of brucellosis.

Objective To monitor the constituents and resistant tendency of bacterial pathogens isolated from diarrheal patients in our hospital form 1994 to 2005 to offer the basis for guiding epidemiologic study,vaccination research and clinical treatment. Methods Enteric pathogenic bacteria were cultured and identified to species,group and serotype with biochemical and serologic methods and the susceptibility of bacteria to antimicrobial agents were tested. Results Enteric pathogenic bacteria were isolated predominantly in male patients and mainly in children and youngsters. It reached a peak from July to September every year. Shigella spp.(75.11%) was the most frequendy isolated pathogens and followed by Vibrio spp.(12.7%),Salmonella spp.(6.28%),Aeromonas spp.(4.43%) and Escherichia coli(1.25%).During the period from 1994 to 2005,diarrheal pathogens had a trend of decrease especially Shigella spp.and Salmonella spp.. Of the 6329 isolates of Shigella spp., 75.62% was S. flexneri and S.soanei,S.dysenteriae and S. boydii constituted 23.98%,0.22% and 0.01% respectively.The sensitivity of different species,group or serotype to different antimicrobial agents was not the same.S.flexneri and Aeromonas spp. were highly resistant to most of antibiotics. However, S.sonnei and Vibrio spp.had good susceptibility to antibiotics tested except trimethoprim/sulfamethoxazole and ampicillin. Conclusion There are many species and serotypes of enteric pathogenic bacteria causing infective diarrhea and the distribution changes gradually in Beijing. The resistance rate of enteric pathogenic bacteria to antibiotics is not the same in different species and serotypes.so strict surveillance iS always needed.

Objective To observe the changes of renal tubular injury and the extent of interstitial fibrosis in the C57BL/6 mouse models of chronic kidney disease(CKD),and provide experimental animal evidence for study of the pro-gression of acute kidney injury(AKI)to chronic kidney disease as well as its mechanisms. Methods Twenty-four 8-week-old male C57BL/6 mice were randomly and equally divided into control group, low-dose, medium-dose, and high-dose cisplatin groups,6 mice in each group. Mice in the cisplatin groups were administrated with 5,7 or 10 mg/kg cispla-tin by intraperitoneal injection once a week for 4 weeks. Plasma creatinine and 24-hour urinary protein were detected to as-sess the renal function. The mice were sacrificed, and plasma and kidney samples were collected for subsequent tests. Pathological changes were observed using periodic acid-Schiff(PAS)staining. To evaluate renal tubules injury, the ex-pression of kidney injury molecule 1(KIM-1)was examined by immunohistochemistry and the level of urinary N-Acetyl-β-D-glucosaminidase was detected with a commercial kit. The infiltration of CD3-positive T cells and F4/80-positive macro-phages was observed by immunohistochemistry(IHC)and immunofluorescence. The expression of collagen I and α-smooth muscle actin(α-SMA)were tested by immunohistochemistry to assess the renal fibrosis, while total kidney collagen was detected by Picrosirius red staining. Results In contrast to the normal control group,the kidney injury became more seri-ous in the cisplatin-treated mice as cisplatin concentration increased. Particularly,significant kidney damage was observed in the high-dose cisplatin group. Compared with the control group,the plasma creatinine and 24-hour urinary protein were significantly increased in the high-dose cisplatin group(P<0.05 and P<0.001)indicating impaired renal function. Mor-phologically,numerous clear vacuoles and necrosis were present in renal tubule epithelial cells in the high-dose cisplatin group. The expression of KIM-1 was markedly up-regulated and the level of urinary NAG was elevated. Infiltration of CD3-positive T cells and F4/80-positive macrophages was enhanced in the mice of high-dose cisplatin group. Data from immuno-histochemistry and picrosirius red staining showed that mice of the high-dose cisplatin group developed renal fibrosis evi-denced by markedly up-regulated expression of collagen I and α-SMA. Conclusions Repeated administration of 10 mg /kg cisplatin for 4 weeks can induce chronic renal insufficiency in mice,which may serve as a novel model for the research on underlying mechanisms of progression from acute kidney injury to chronic kidney disease.

Objective To investigate the method and effect of organization and management on self -protection among nursing staff of Chinese Military Medical Aid Team to Sierra Leone , so as to provide a reference for self-protection management among the nursing staff who will be dispatched to Africa for fighting against Ebola virus .Methods The organization and management of self-protection included enhancing the awareness of self-protection , standardizing protection procedures , perfecting rules and regulations , and strengthening physical and mental health care .Results A total of 274 patients were admitted to the holding center , of which 145 were confirmed as Ebola virus disease , and there was no Ebola virus infected nursing staff in our team .The organization and management of self-protection among nursing staff served as an important guarantee for prevailing over Ebola virus .Conclusions It is crucial to provide a scientific and effective organization and management of self-protection to prevent Ebola virus infection for nursing staffs .

Objective To explore the application effect of scenario simulation training to prevent and control Ebola virus disease spread among cleaners in Sierra Leone .Methods A total of forty local cleaners in Sierra Leone were given scenario simulation training program designed to resolve the problems of Ebola infection prevention and control , and they were required to be appraised by corresponding criteria after drilling and training.Results After training, all cases were significantly improved in anxiety , tense, fear, agrypnia and depression.There were statistical difference before and after training (χ2 =16.2,39.7,34.3,15.9,5.2, respectively;P<0.05).Seven of forty cleaners had satisfaction with environment layout modification before training compared with thirty nine of forty having satisfaction after training (χ2 =52.4,P<0.01).100% of participants presented the satisfaction of training contents and effects .The highest score of examination was 98 and lowest was 68 that 39 of them reached passed level .All cleaners showed excellent working status and there was no infections happened .Conclusions It is an effective way to take step-by-step and whole-procedure scenario simulation training to increase the knowledge and skills of infection prevention and control .