Objective To investigate the carbapenem-resistant mechanism of one strain of Klebsiella pneumoniae( KPN) and one strain of Pseudomonas aeruginosa( PAE) .Methods The identity of the isolates was confirmed by using MALDI-TOF mass spectrometry, 16S rDNA and special gene amplification .The minimum inhibitory concentrations (MICs) of the antimicrobial agents were determined by VITEK 2 Compact System .CarbaNP Antimicrobial susceptibility testing , plasmid extraction, electroporation experiment , PCR amplification, and cloning and sequencing were carried out to analyze the en-coding gene of β-lactamases.Results The types of β-lactamases of the KPN were blaKPC-2 and blaSHV, confirmed by se-quencing of the PCR products , and that of the PAE were blaKPC-2 .Only blaKPC-2 was displayed in both transformants .All of the results of CarbaNP were type A .Conclusion Both strains of KPN and PAE resisting to carbapenem produce a plasmid-mediated carbapenemase blaKPC-2 , which belongs to Bush group 2f, class A β-lactamase.The extended-spectrum β-lacta-mases gene encoding blaSHV of the KPN might be located in the chromosome or not in the plasmid carrying with blaKPC-2 .

Objective To investigate the use of glucocorticoids in patients with severe community-acquired pneumonia (SCAP) in the intensive care unit (ICU) of Hospitals in Zhejiang Province and to provide a reference for guiding clinical use of SCAP patients. Methods To draw up a questionnaire with reference to the Chinese and international guidelines, and to investigate the knowledge of community-acquired pneumonia (CAP) related guidelines and the use of glucocorticoids in patients with SCAP by doctors in hospitals above secondary level in Zhejiang Province by Email. Then the valid questionnaire was analyzed. Results In June 2016, 340 questionnaires were distributed, and all were returned after 2 months, with 333 of valid; 333 doctors from 45 ICUs in Zhejiang Province participated in the survey. ① The knowledge of CAP-related guidelines in ICU doctors: 79.58% (265/333) of the doctors had read the CAP guidelines, and those who work over 10 years had a higher reading rate than those with 1-5 years and 6-10 years [93.07% (94/101) vs. 74.00% (111/150), 73.17% (60/82), both P < 0.05]. Post-graduates and above had higher reading rates than undergraduates [85.35% (134/157) vs. 74.43% (131/176), P < 0.05]. Senior doctors had higher reading rates than the junior and intermediate doctors [93.07% (94/101) vs. 71.43% (80/112), 75.83% (91/120), both P < 0.05]. The rate of understanding the clinical application of glucocorticoids was 13.81% (46/333). The doctors who work over 10 years and the seniorshad a relatively high awareness rate, 23.76% (24/101) and 20.79% (21/101) respectively. However, there was no significant difference in the awareness rate between doctors with different degrees and different levels of hospitals. ② For the use of glucocorticoids in different causes of pneumonia, 44.74% (149/333) of doctors routinely used glucocorticoids in severe viral pneumonia. The proportion of glucocorticoids used in severe bacterial pneumonia, severe fungal pneumonia, severe pneumocystis pneumonia, chronic obstructive pulmonary disease (COPD) and severe pneumonia were 22.82% (76/333), 9.31% (31/333), 22.52% (75/333) and 18.32% (61/333), respectively. ③ The way of glucocorticoid usage: 79.58% (265/333) of doctors chose methylprednisolone, 4.20% (14/333) chose hydrocortisone, 1.20% (4/333) chose dexamethasone, and 15.02% (50/333) had not use glucocorticoids. The proportion of physicians who chose to use glucocorticoids within 24 hours of admission and 1-7 days after admission were 52.65% (149/283) and 47.35% (134/283), respectively. Glucocorticoids were used more in doctors with lower academic qualifications and hospitals within 24 hours. The undergraduate degree was 61.39% (97/158), and the second-grade class hospital was 67.50% (27/40). Among the doctors who chose methylprednisolone, 60.75% (161/265) prescribe the dose ≤80 mg/d;79.15% (224/283) chose the course of ≤7 days. The number of years of work, education, professional title and hospital grade had no significant effect on the choice of methylprednisolone and the course of treatment. Conclusions ICU doctors of 45 hospitals in Zhejiang Province have a high degree of heterogeneity in the understanding of the use and guidelines of glucocorticoids in SCAP. It is necessary to strengthen the ICU doctor's study of clinical guidelines at home and abroad and to develop a glucocorticoid use plan according to the specific conditions of patients, so that SCAP patients can benefit more.