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【Objective:】 To construct the training index system of humanistic care for nursing students, and to provide reference for optimizing the educational model of humanistic care. 【Methods:】 Taking the quality care model as the theoretical framework, through the literature analysis, semi-structured interviews, and Delphi expert consultation, the nursing students’ humanistic care training index system was initially constructed, and the weights were determined by the analytic hierarchy process. 【Results:】 After two rounds of expert consultation, the expert authority coefficients were 0.89 and 0.91, the effective recovery rates of the questionnaires were 91.30% and 95.24%, respectively; the Kendall’s W values of expert correspondence were 0.179 and 0.118, respectively (P<0.05); The mean value of each index was 4.45~4.95, the coefficient of variation was 0.05~0.17, and the full score rate was 50%~95%. The humanistic care training index system for nursing students included 5 first-level indicators and 30 second-level indicators. 【Conclusion:】 Based on Delphi method and analytic hierarchy process, the training index system of humanistic care for nursing students has high enthusiasm, authority, concentration and coordination of experts, which is scientific, reliable and practical, and can provide reference for the development of caring education in colleges and universities.
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OBJECTIVE@#To establish a quantitative fluorescent detection method using DAPI for detecting inorganic polyphosphate (polyP) in enterohemorrhagic Escherichia coli (EHEC) O157:H7.@*METHODS@#The DNA of wild-type strain of EHEC O157:H7 was extracted and purified. DAPI was combined with the extracted DNA and polyP45 standards for measurement of the emission spectra at 360 nm and 415 nm fluorescence spectrophotometry. The fluorescence of DAPI-DNA and DAPI-polyP complexes was detected by fluorescence confocal microscopy to verify the feasibility of DAPI for detecting polyP. To determine the optimal pretreatment protocol for improving the cell membrane permeability, the effects of 6 pretreatments of the cells (namely snap-freezing in liquid nitrogen, freezing at -80 ℃, and freezing at -20 ℃, all followed by thawing at room temperature; heating at 60 ℃ for 10 min; treatment with Triton x-100; and placement at room temperature) were tested on the survival of EHEC O157:H7. The fluorescence values of the treated bacteria were then measured after DAPI staining. A standard calibration curve of polyP standard was established for calculation of the content of polyP in the live cells of wildtype EHEC strain and two mutant strains.@*RESULTS@#At the excitation wavelength of 360 nm, the maximum emission wavelength of DAPI-DNA was 460 nm, and the maximum emission wavelength of DAPI-polyP was 550 nm at the excitation wavelength of 415 nm. The results of confocal microscopy showed that 405 nm excitation elicited blue fluorescence from DAPIDNA complex with the emission wavelength of 425-475 nm; excitation at 488 nm elicited green fluorescence from the DAPIpolyP complex with the emission wavelength of 500-560 nm of. Snap-freezing of cells at -80 ℃ followed by thawing at room temperature was the optimal pretreatment to promote DAPI penetration into the live cells. The standard calibration curve was =1849+127.5 (R=0.991) was used for determining polyP content in the EHEC strains. The experimental results showed that wild-type strain had significantly higher polyP content than the mutant strains with deletion.@*CONCLUSIONS@#We established a convenient quantitative method for direct and reliable detection polyP content to facilitate further study of polyP and its catalytic enzymes in EHEC O157:H7.
Subject(s)
Escherichia coli O157 , Escherichia coli Proteins , PolyphosphatesABSTRACT
<p><b>OBJECTIVE</b>To construct enterohemorrhagic Escherichia coli (EHEC) O157:H7 strains with delection espF gene and its nucleotide fragment and with espF gene complementation.</p><p><b>METHODS</b>A pair of homologous arm primers was designed to amplify the gene fragment of kanamycin resistance, which was transformed into EHEC O157:H7 EDL933w strain via the PKD46 plasmid by electroporation. The replacement of the espF gene by kanamycin resistance gene through the PKD46-mediated red recombination system was confirmed by PCR and sequencing. The entire coding region of espF along with its nucleotide fragment was amplified by PCR and cloned into pBAD33 plasmid, which was transformed into a mutant strain to construct the strain with espF complementation. RT-PCR was used to verify the transcription of espF and its nucleotide fragment in the complemented mutant strain.</p><p><b>RESULTS AND CONCLUSION</b>We established EHEC O157:H7 EDL933w strains with espF gene deletion and with espF gene complementation. Both espF and its nucleotide fragment were transcribed in the complemented mutant strain. The two strains provide a basis for further study of the regulatory mechanism of espF.</p>
Subject(s)
Carrier Proteins , Genetics , DNA Primers , Escherichia coli O157 , Genetics , Escherichia coli Proteins , Genetics , Gene Deletion , Plasmids , Polymerase Chain ReactionABSTRACT
Objective: To research three-dimensional computed tomography angiography (3D-CTA) combined three-dimensional digital subtraction angiography (3D-DSA) in the diagnosis of intracranial aneurysms. Methods: 3D-CTA and 3D-DSA were performed in 27 cases with Highly suspected in patients with intracranial aneurysms, And compared with the final outcome of surgery. To evaluate 3D-CTA and 3D-DSA in the detection rate of aneurysms, three-dimensional shape and spatial relations ability to display. Research 3D-CTA combined 3D-DSA in the diagnosis of intracranial aneurysms. Results: Twenty-seven cases of suspected patients, 25 cases of surgical treatment of confirmed aneurysms, 28 aneurysms found. 3D-CTA and 3D-DSA rate of aneurysms was no significant difference. 3D-CTA of the aneurysm can be accurately detected clearly shows its three-dimensional shape and spatial relationship with the surrounding skull, 3D-DSA for aneurysm neck, the tumor and the aneurysm and parent artery relationship shows clear and accurate. Conclusion: Imaging of intracranial aneurysm diagnosis, 3D-CTA and 3D-DSA have their respective advantages, combined two methods can improve the detection rate of aneurysms. 3D-CTA combined 3D-DSA have support of design for operation.
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Objective To investigate the satisfaction degree with nursing for discharged patients by telephone interview,and understand the nursing quality improvement during hospitalization of patients.Methods In January and December 2011,24 clinical departments were selected as the research object,10 patients were selected from each department.The questionnaires of satisfaction degree with nursing were adopted to investigate the satisfaction degree of patients by telephone interview.The difference of satisfaction degree with nursing were compared between January and December 2011.Results Compared with the results of January,there were statistical differences in overall mean score of satisfaction degree and the dimensions of service attitude,knowledge information,ward management and working ability.While the means of dimensions of basic nursing care and care-taking patient were in high levels.Conclusions The method of telephone interview to investigate the satisfaction degree of discharged patients is direct,real and objective.It is convenient for the hospital to understand the nursing quality and existing problems during hospitalization of patients,and offers scientific way for continuous improvement of nursing service.