Objective To explore the role of ADSCs and PRP in soft tissue defect repairing .Methods Harvest adipose tissue from inguinalis fat pad of SD rats ,isolation ,culture ,and identification the ADSCs through three differentiation method .Take 30 male SD rats about 6-7 weeks old randomly .Randomly selected 12 rats been take blood from heart .Preparation the PRP with modi-fied appel method .To count platelet of whole blood and PRP under microscope .Take the remaining rats .Divided the rat into 3 groups(n=6 in each group) randomly ,of which group A treatment with ADSCs and PRP ;Group B treatment with ADSCs ;Group C treatment whit PRP .Selected one side of skin defect wound for test randomly and the relative side skin defect is for control ,Handle all of the control wound to group D ;Observe the grow th of granulation tissue on the wound surface ;observe the inflammatory sur-rounding and the degree about epithelial .statistical analysis and record the wound size ,and calculation the shrinkage rate of wound in different periods .Record the time of completely healing time .Histologic observation of the wound healing tissue .Results Plate-let counting showed platelet of PRP is 5 .21 times than whole blood .The wound completely healed time :group A (18 .25 ± 1 .44 ) days ,group B(19 .13 ± 1 .28)days ,group C(19 .72 ± 0 .87)days ,group D(22 .31 ± 1 .65) days ,The time of each treatment group and control group was significantly obvious(P< 0 .05) .At 3 ,7 ,11 and 15 days after experimental treatment ,compared with the control group the experiment group of wound contraction rate was significantly obvious (P<0 .05) .Conclusion Application of AD-SCs with PRP can enhance the quality and shorten the wound healing time than used them alone .

Objective To investigate the species and drug resistance of pathogens causing bloodstream infections in hospitalized patients,and provide scientific evidence for antimicrobial use and control of healthcare-associated blood-stream infection.Methods From January 1 to December 31,2012,16 428 blood specimens were performed blood culture,pathogens were isolated and performed antimicrobial susceptibility testing.Results Of 16 428 blood speci-mens from 5 546 patients,384 (6.92%)were positive for blood culture,398 pathogenic isolates were detected,of which gram-positive bacteria,gram-negative bacteria,and fungi accounted for 23.62% (n=94),68.34% (n=272),and 8.04% (n=32)respectively,positive rate of blood culture were highest in 61-80 age group(8.26%), the top five departments of positive rate of blood culture were departments of burn,traditional Chinese medicine, cardiac intensive care unit,transplantation and traumatology;gram-positive cocci were highly susceptible to vanco-mycin,teicoplanin and linezolid,one Enterococcus faecium strain was found to be resistant to vancomycin;Among gram-negative bacilli,Enterobacteriaceae were highly susceptible to amikacin and carbapenems;drug resistance rates of Acinetobacterbaumannii and Pseudomonasaeruginosa to carbapenems was 70.97% and 35.90% respective-ly.Conclusion Gram-negative bacteria are the major pathogens causing bloodstream infection,positive rate of blood culture of elderly people is high.It is necessary to conduct regular surveillance on distribution and drug resistance of pathogens.

Objective Methicillin-resistant Staphylococcus aureus (MRSA) is a growing public health concern that has been associated with pediatric fatalities. This study investigated the genotypes of staphylococcal cassette chromosomal mec (SCCmec) and Panton-Valentine Leukocidin (PVL) in MRSA strains isolated from Shanghai Children's Hospital by PCR. Methods A total of 30 strains of MRSA were isolated from various clinical specimens from October 2005 to June 2006. The antimicrobial susceptibility was measured by agar diffusion method. SCCmec typing was conducted using a novel multiplex PCR assay allowing for concomitant detection of methicillin resistance (mecA gene) to facilitate detection and classification of all currently described SCCmec typesⅠ, Ⅱ, Ⅲ, Ⅳa, b, c, d andⅤ. PVL gene was also determined by PCR. Results mecA gene was positive in all the strains. SCCmecⅡ was identified in 6(20.0%) isolates, SCCmecⅢ in 15(50.0%) isolates, SCCmecⅤ in 2 and SCCmecⅣa in 1 isolate. Six MRSA strains were non-typeable. The isolates with SCCmecⅡ or SCCmecⅢ were resistant to multiple antibiotics. The strains harboring SCCmecⅣa or SCCmecⅤwere susceptible to all antibiotics except β-lactams. Eleven (36.7%) isolates were PVL positive. The genotypes and subgenotypes of staphylococcal chromosomal cassette mec of eleven PVL-positive MRSA were SCCmecⅡ(1 isolates), SCCmecⅢ (5 isolates), SCCmecⅣa (1 isolate), SCCmecⅤ (2 iso-Lates) non-typeable (2 isolates). Conclusions SCCmecⅡ and SCCmecⅢ are the major genotypes of MRSA in our hospital. These isolates are multi-resistant to antibiotics. The prevalence of PVL gene is higher in SCCmecⅡ- or SCCmeⅢ-positive MRSA. The isolates with SCCmecⅡ or SCCmecⅢ were resistant to multiple antibiotics.

Objective·To analyze changes in the type distribution and drug resistance of pathogenic bacteria isolated from burn wards and to provide evidence for rational use of antibiotics, reduction of drug-resistant isolates, and hospital infection control. Methods·Isolates from burn patients were collected from January 2011 to December 2014. Statistical analysis of infection sources, type distribution, and changes in resistance rates of main pathogens during the four year period was performed. Results·A total of 2399 isolates were collected, including 1286 (53.61%) gram-negative bacilli (G-b), 1088 (45.35%) gram-positive cocci (G+c), and 25 (1.04%) fungi. The most common G-b pathogens were Pseudomonas aeruginosa (447, 34.76%) and Acinetobacter baumannii (369, 28.69%). The most common G+c pathogen and fungus were Staphylococcus aureus (489, 44.94%) and Candida albicans (8, 33.33%), respectively. In the last two years, the detection rates of S.aureus and A.baumannii were significantly lower and the detection rate of P.aeruginosa was significantly higher than those in the first two years (P<0.05). P.aeruginosa and A.baumannii showed high resistance (>80%) to the third and fourth generation cephalosporins, carbapenems, aminoglycosides and quinolones, but the changes were not statistically significant (P>0.05). S.aureus was only highly resistant to penicillin (97.58%) and was 100% susceptible to vancomycin. Its resistance rates toward cefazolin, ampicillin/sulbactam, gentamicin, levofloxacin, and rifampin decreased significantly (P<0.05). The detection rate of methicillin-resistant S. aureus (MRSA) dropped from 72.28% to 63.00%. Conclusion·Many types of drug resistant bacteria were detected in burn wards. The drug resistance problem was serious. Improving management and rational use of antibiotics can reduce the occurrence of drug-resistant bacteria and increase the efficacy of clinical anti-infective treatment and nosocomial infection control.

OBJECTIVE To investigate the distribution and antibiotic susceptibility of pathogens causing urinary infection,for the guide of rational use of antimicrobial agents in clinic. METHODS The bacteria isolated from the middle segment urine sample were identified by ATB system,and K-B method was used to study the antimicrobial resistance.The data were analyzed by WHONET 5.3. RESULTS Escherichia coli was one of the most common bacteria in the urinary tract infection(57.6%),and then were Enterococcus(14.4%).The results of antibiotic susceptibility test in vitro showed the susceptibility rate of Enterobacteriaceae and Acinetobacter baumannii to imipenem,was 100.0%,but the resistance rate of Pseudomonas aeruginosa was 13.6%.Gram-positive cocci were sensitive to glycopeptide antibiotics and linezolid. CONCLUSIONS Clinician should pay attention to the kinds of pathogenic bacteria causing urinary tract infection and their susceptibility to clinically common used antibiotics for reasonable use of drugs.

OBJECTIVE To investigate the ESBLs produced by clinical strains of Enterobacter cloacae.METHODS Production of ESBLs was identified with modification of the double-disk test(MDDT).Eight kinds of primers(CTXM,CTXM-1,CTX-M-2,CTX-M-9,SHV,TEM,VEB,and PER)were used for the PCR amplification.Gene clone and DNA sequencing were performed then.RESULTS The result of MDDT was positive,amplicons were gained by PCR amplification with CTXM-1.DNA sequencing of amplicons of this strain revealed ESBLs of CTX-M-3.CONCLUSIONS ESBLs are the important reason for E.cloacae resisting to the third-generation cephalosporins.

Objective To evaluate the capability of three tests used alone or in combination for identification of Staphylococcus aureus.Methods Identification of Staphylococcus aureus by the detection of spa gene with PCR and the Vitek-2 system were selected as the reference methods.Comparison of three phenotypic tests including DNase,mannitol fermentation and tube coagulase test was carried out to analyze the sensitivity,specificity,positive/negative predictive value,positive/negative likelihood ratio and Youden index.The consistency,cost and related indexes of the assays were analyzed between the combined phenotypic tests and the reference methods.Results In the present study,324 isolates of Staphylococci,including 293 Staphylococcus aureus and 31 non-Staphylococcus aureus,were collected.Single biochemical test could not identify Staphylococcus aureus efficiently.Comparison between the reference methods and the combined three biochemical tests by Kappa statistic analysis indicated that an overall Kappa value was 0.9441,and the algorithm of combined test was less costly.The sensitivity and specificity of this algorithm were 100％ and 90.3％,respectively.Conclusion The cost-effective algorithm of combined DNase,mannitol fermentation and tube coagulase test could efficiently distinguish Staphylococcus aureus from other Staphylococcus species.

Objective To investigate the relationship between antibiotic use and antimicrobial resistance in Acinetobacter bau-mannii for rational use of antibiotics.Methods Antibiotic use density (AUD)of common antibiotics in hospitalized patients were collected in a tertiary hospital between 2006 and 2010.Clinical isolates of A.baumannii from those patients were collect-ed.The resistance to common antimicrobial agents were tested by disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI,2012)guidelines.Relationship between antibiotic use and antimicrobial resistance in A.baumannii was analysed by SPSS 16.0.Results The resistant rates of A.baumannii isolated from inpatients were high.Consumption of cephalosporins and quinolones were large.There was a positive correlation between the resistant rate of A.baumannii to imi-penem and AUD of carbapenems (r=0.975,P <0.05).The resistant rate of A.baumannii to meropenem showed significantly positive relation to AUD of carbapenems (r= 0.975,P <0.05).Resistant rates of aminoglycosides,quinolones,cephalospo-rins and beta-lactamase inhibitors was not correlated to AUD of those antibiotics.Conclusions We should pay more attention to the high prevalence of resistant A.baumannnii strains.Application of imipenem and meropenem should be strictly controlled.Amikacin and beta-lactamase inhibitors are better choice for empirical antibiotic therapy in the treatment of infections caused by A. baumannii.

Objective To evaluate the practicability of CHROMagar orientation medium combined with simple biochemical tests for identification of common oxidase-negtive gram-negative bacilli.Methods The CHROMagar orientation medium was used together with biochemical tests including indole test , ornithine decarboxylase test and lysine decarboxylase test for identification of common oxidase -negtive gram-negative bacilli.The sensitivity, specificity, likelihood ratio, Youden index and Kappa value of the diagnostic assays were evaluated .McNemar test was performed to evaluate facticity, accuracy and cost of the method in com-parison with the Vitek-2 system as reference method .Results The identification of oxidase-negtive gram-negative bacilli from 318 bacterial strains showed that the sensitivities and specificities of CHROMagar orien-tation mediumm in combination with simple biochemical tests to Serratia marcescens, Stenotrophomonas mal-tophilia and Acinetobacter baumannii reached 100%, and for Escherichia coli, Enterobacter aerogenes and Klebsiella pneumoiae were above 90%.The specificities for identification of Enterobacter cloacae, Klebsiella oxytoca, Citrobacter freundii and Proteus mirabilis were all above 90%, but the sensitivities were around 75%-90%.Kappa values of the assays were above 0.85, howerer, which was only 0.5947 for Citrobacter freundii.McNemar test showed that all P values were above 0.05, and cost of the assays was reduced by 90%.Conclusion CHROMagar orientation medium in combination with simple biochemical tests is a cost-effective assay for identification of common oxidase-negtive gram-negative bacilli .

Objective To investigate the production of carbapenemase and 16S rRNA methylase in five isolates of pan-drugs resistant E.cloacae recovered in Ruijin hospital.Methods MICs of the five isolates to 10 antibiotics were determined by E test.Six kinds of 16S rRNA methylase genes and a series of β- lactamase genes were amplified by PCR.Shotgun cloning was performed to detect carbapenem resistance determinant.The conjugal transfer of carbapenemase gene and 16S rRNA methylase gene was performed in broth culture with E.coli J53 as the recipient.Pulsed-field gel electrophoresis (PFGE) was carried out to analyse the genotyping.IEF was performed to detect β-1actamases.Southern blot was performed to determine the location of carbapenem resistance determinant.Results The MICs of 10 antibiotics were ＞32 mg/L.Four β-1actamases with pIs of 5.4 ( TEM-1 ),6.7 ( KPC-2 ),8.2 ( SHV-12 ),8.4 (CTX-M-14) were determined.The insertion sequence in the recombinant plasmid was blaKPC-2 flanked by a transposon.blaKPC-2 was located on a large non-conjugative plasmid whereas armA was located on an other conjugative plasmid.PFGE patterns of 5 isolates were identical.Conclusion KPC-2 was responsible for carbapenem resistance in pandrugs resistant Enterobacter cloacae.There was no relationship between blaKPC-2 and armA.Although pandrug resistant Enterobacteriaceae remain rare,the emergence of this group of organism merits monitoring.