OBJECTIVE To investigate the clinical distribution and antimicrobial resistance of infections caused by Enterobacter cloacae in our hospital,for guiding the prophylaxis and treatment of these infections in clinical practice. METHODS Antimicrobial susceptibility tests were done by Kirby-Bauer disk diffusions method,the phenotypes of E.cloacae strains harboring AmpC ?-lactamases and extended-spectrum ?-lactamases(ESBLs) were detected by modified three-dimensional extract test. RESULTS A total of 162 strains of E.cloacae were isolated between Jan 2002 and Dec 2004,the percentages of these strains isolated from sputum,urine and wound secretion were 62.3%,11.7% and 9.9%,respectively.73.5% of all strains isolated from intensive care unit(ICU) in every endemic area.AmpC ?-lactamases,AmpC ?-lactamases combined with ESBLs and ESBLs producing strains were detected in 19.1%,11.7% and 14.2% of E.cloacae,respectively.The resistance of AmpC ?-lactamases or ESBLs producer was obviously higher than that of non-AmpC ?-lactamases and ESBLs producer,while the phenomenon of resistance was serious especially in AmpC ?-lactamases combined with ESBLs producer,but all strains were sensitive to imipenem. CONCLUSIONS E.cloacae causes infections of respiratory tract,urinary tract and wound principally in clinic,the most of which occurred in ICU.AmpC ?-lactamases and ESBLs are popular in E.cloacae,the restricted use of ?-lactams is a considerable measure which reduces prevalence of AmpC(?-lactamases) and ESBLs producing strains.The antimicrobial agents for treating infections caused by E.cloacae should be chosen according to the results of the antimicrobial susceptibility tests,imipenem is the first choice for severe infections.

Objective To study the effects of different inner face status of human venous blood container on platelet activation. Methods The plastic( polyethene terephthalate,PET) and glass tubes were coated with polyalkyleneoxide modified polydimethylsiloxane(L722). The contact angles of L722-coated glass and L722- coated PET tubes, glass tubes, PET tubes, silane coupling agent-coated glass tubes and polypropylene (PP) tubes were analyzed respectively. The blood were drawn into above tubes, and then incubated in a roller bottle at room temperature for 10-60 mira The marker of activated platelete, CD62p, was detected by flow cytometry(FCM). Results The inner face contact angle of the blood collection tubes with different material and surface treatment reflected platelet activation to a certain extent, but was not linear. The percent age of CD62p positive platelets in L722-modified PET tubes reduced from (37.4 ±14. 8) % to(21.9 ± 12. 4) %. The platelet activation by glass tubes was (54.5 ± 18.6 ) %, markedly more than PET tubes. While membrane formed, the platelet activation by glass tubes decreased remarkably, and the percentage of CD62p positive platelet in silane coupling agent-coated glass tubes were ( 28. 3 ± 8.2 ) %,markedly less than that of the L722-coated glass tubes. The platelet activation by PET-based material ectally modified with L722 was obviously less than L722-coated glass tubes and the percentage of CD62p positive platelet in silane coupling agent-coated glass tubes were ( 41.5 ± 15.9 ) % and ( 22. 0 ± 12. 8 ) %,respectively. The time course of platelet activation by different tubes showed that the platelet activation by L722-coated PET tubes and polypropylene tubes in 60 rain was not significantly different from the results in30 mitt Conclusions The diverse surface energy status induced by different material and surface treatment of blood collection tubes have obvious effects on the activation of platelets. The silicic oil surface treatment can effectively improve the blood compatibility of blood collection tubes. CD62p detectod by FCM is a sensitive marker for the evaluation of the activation of platelets induced by the material of blood collection tubes. It is of importance in the establishment of surface treatment model of blood container and screening out the material of clinical application.

Objective To study the mutations of Pseudomonas aeruginosa oprD gene in Imipenem-resistant clinical isolates. Methods Random amplified polymorphic DNA typing(RAPD) was carried out to analyse the homology in 10 Imipenem-resistant clinical isolates. Pseudomonas aeruginosa oprD gene in 10 Imipenem-resistant clinical isolates were amplified by polymerase chain reaction and sequenced. Results Ten Imipenem-resistant clinical isolates were divided into four different clones which can not produce metallo ?-lactamase. As compared with sequence X63152, oprD gene of Pseudomonas aeruginosa varied greatly. The aberration rates exceed 50%. There were multiple point mutations within 276～387 bp coding region of 30, 11, 9, 20, 31 strains. Due to the mutations of 308 bp G→C and 344 bp C→A, threonine and diaminocaproic acid were replaced by serine and threonine respectively. The DNA deletion of 393～412 bp in oprD gene contributes to the frame shift mutation in the following nucleotide sequence. The deletion of 264～273 bp in the coding region of oprD gene in 13 and 21 clone strains leads to frame shift mutations forming terminal codon TAA(319～321 bp).Base substitutions and multiple point mutations were obvious in the coding regions of 1 and 22 clone strains. Their aberration rates were 54.03% and 74.89% respectively. Conclusions The mutations of Pseudomonas aeruginosa oprD gene in Imipenem-resistant clinical isolates are various.

Objective To explore the effect of miniAd-ATP7B-GFP on the copper metabolism of skin fibroblasts of Wilson′s disease ( WD ) patients under high concentration copper medium.Methods Firstly, mini-adenovirus carrier containing human ATP7B gene was built and the mutations of 8 WD patients were detected.Fibroblasts from primary culture of skin of WD patients and normal human were cultivated 72 h in basic medium and medium with the copper concentration of C1(22.3μmol/L), C2(89.2μmol/L), C3 (156.1 μmol/L), C4 (245.3 μmol/L).Then the concentration of copper and protein was detected and copper/protein ratio was calculated.Secondly, miniAd-GFP ( miniAd-GFP group) and miniAd-ATP7B-GFP ( miniAd-ATP7B-GFP group ) were added into WD patients skin fibroblasts respectively, Wilson non-transfection group and normal group were set up as control, and C4 medium was used to culture the cells of four groups for 72 h and 96 h.Then the concentration of copper and protein was detected and copper/protein ratio was calculated.Results Five kinds of mutations were detected from 8 WD patients.The copper/protein ratio of WD patients and normal human in basic medium and the C1 -C3 groups had no statistically significant difference, but in C4 group (WD (1 871.6 ±209.2) ng/mg, normal group (1 267.2 ±188.3) ng/mg) the difference was statistically significant (t=6.075, P<0.01).C3((816.3 ±113.9) ng/mg) and C4 groups had statistically significant difference compared with the basic medium group ( ( 159.2 ± 38.6) ng/mg;WD:χ2 =31.493, normal group:χ2 =30.708, both P<0.01).The copper/protein ratio of 96 h group was higher than 72 h group.Compared with WD non-transfection (96 h:(2 731.2 ±188.7) ng/mg,72 h:(1 901.7 ±219.5) ng/mg) and normal groups, miniAd-ATP7B-GFP group had statistically significant difference both in 96 h ( ( 2 071.0 ±171.8 ) ng/mg ) and 72 h groups ( ( 1 495.5 ±161.4 ) ng/mg;72 h:F=20.130, 96 h: F=51.496,P<0.01).Conclusion MiniAd-ATP7B-GFP has partial improvement on copper metabolism of skin fibroblasts of WD patients under high concentration copper medium.

Objective To observe the effect of psychological intervention on nosohemia patients during hematopoietic stem cell collection technique. Methods 44 perioperative nosohemia patients were assigned to experiment group and control group. The experiment group took a psychological intervention. Results Anxiety and depression in the experiment group improved better than in the control group (P<0.05) while there was no significant difference in anxiety and depression in the control group after psychological intervention (P>0.05). Conclusion Psychological intervention could effectively reduced the negative emotion of perioperative nosohemia patients during hematopoietic stem cells collection.

OBJECTIVE To study the antimicrobial susceptibility and molecular epidemiology of imipenem-resistant Pseudomonas aeruginosa isolated from patients in burn wards in a hospital of Guangzhou. METHODS A total of 48 imipenem-resistant isolates were collected,antimicrobial susceptibility was tested by KirbyBauer disk diffusion method.Random amplified polymorphic DNA(RAPD) typing was carried out to analyze the homology. RESULTS All isolates were multiresistant,the orders of sensitivity rates of antibiotics were CIP,TOB,AMK,GEN,and FEP.Forty eight strains were classified into types A,B,C,and D based on RAPD pattern,types A and B were the most epidemic clones. CONCLUSIONS The prevalence of imipenemresistant P.aeruginosa in burn wards of the hospital is due to nosocomial infection.The prevalent strains are multiresistant.

OBJECTIVE To analyze homology,antimicrobial susceptibility and metallo-?-lactamase gene type of metallo-?lactamase producing Pseudomonas aeruginosa in burn wards.METHODS Antimicrobial susceptibility tests were performed with E-tests.Using 2-mercaptoethanol disc synergy test to screen metallo-?-lactamase(positive) strains from imipenem-resistant P.aeruginosa in burn wards.Metallo-?-lactamase gene and integrase gene were amplified and sequenced.Resistance plasmid transfer and curing experiments were implemented to study the transfer of imipenem resistance and plasmid DNA was extracted and purified with Qiagen Plasmid Mini Kit.Random amplified polymorphic DNA(RAPD) typing was carried out to analyze the homology of the isolates.(RESULTS) Six strains of P.aeruginosa were suggested to produce metallo-?-(lactamase) by 2(-mercaptoethanol) disc synergy test.Using primers described for bla(VIM),the amplifications were observed among all 6 isolates and a VIM-2 metallo-?-(lactamase) gene was identified by sequencing.All isolates which producing VIM-2 metallo-?-(lactamase) had class 1 integrase gene and derived from a same clonal origin.CONCLUSIONS(VIM-2) metallo-?-(lactamase) producing P.aeruginosa is prevalent in burn wards,all isolates which producing VIM-2 metallo-?-(lactamase) have class 1 integrase gene.

Nasopharyngeal carcinoma (NPC), a common head and neck neoplasm in southern China,is mostly treated with radiotherapy, however, it can not be completely cured by chemoradiotherapy. Recent investigation on NPC suggests that the nasopharyngeal carcinoma stem cells can be differentiated to heterogeneous cancer cells that have a strong ability for self renovation and further contribute to the development and progression of tumor itself which is closely related to drug resistance, recurrence and metastasis of NPC. Wnt/β-catenin, Notch, Hedgehog, NF-κB and mTOR signaling pathways play important roles in cancer stem cells. Study on nasopharyngeal carcinoma stem cells targeted therapies and its related pathways provides a new strategy for the clinical treatment of NPC.

Objective To construct an objective,comprehensive core competence evaluation index system for nursing postgraduates,and to provide evidence for postgraduates nursing education.Methods According to literature review,expert interviews,focus group discussions,Delphi and other research methods,we conducted two rounds of 32 experts consultation in 18 cities from 10 provinces to establish the final core competence evaluation index system.Results The two experts consultation derived core competence evaluation index system including 7 level-1 indexes,20 level-2 indexes and 53 entries.Level-1 indexes:theoretical knowledge and skills,critical thinking ability,research ability,clinical care ability,teaching skills,interpersonal communication skills,professional development skills.Conclusions This program is rigorous and reliable,with some theoretical and practical significance of its results,which provide a reference for the training program,evaluation of the quality,assessment of postgraduate nursing education.

ObjectiveTo explore the effect of sustained spinal cord compression on spinal cord injury (SCI).MethodsTwenty four dogs underwent sustained spinal cord compression for 30 minutes (group A), 90 minutes (group B) and 180 min (group C) with a device designed by the authors for weight loading spinal compression, and anthor four dogs as the control group (group D) only underwent surgical operation without SCI. Cortex somatosensory evoked potentials (CSEP) were monitored during all the procedure of making model. The changes of pathologic histology, imaging and behavior examination of all animals were observed.ResultsThe lesion degrees gradually increased from group A to group C (P<0.01), and there was similar outcome in MRI imaging. The behavior improvement in group A was more significantly than that in groups B and C (P<0.05).ConclusionSustained spinal cord compression aggravates the extent of SCI process, and the decompression must be taken as soon as possible.