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OBJECTIVES@#The prevalence of carbapenem-resistant Enterobacterales (CRE) presents a significant challenge in clinical anti-infective treatment. This study aims to investigate drug resistance and the molecular epidemiological characteristics of CRE in our area. Additionally, we seek to evaluate practicality of utilizing carbapenemase inhibitor enhancement test in clinical laboratory.@*METHODS@#Non-repeated CREs isolated from clinical specimens at Xiangya Hospital, Central South University, were collected. Minimum inhibitory concentration (MIC) combined with Kirby-Bauer (KB) assay was used to detect the drug susceptibility of the strains, and 13 carbapenemase-producing genes were detected by PCR. The phenotype of 126 strains of carbapenemase-producing Enterobacterales identified by PCR was detected by the carbapenemase inhibitor enhancement test to understand the agreement between the method and the gold standard PCR results.@*RESULTS@#Among 704 CRE strains examined, we observed significant drug resistance in 501 strains dentified as carbapenemase-producing Enterobacterales (CPE). Klebsiella pneumoniae was the predominant CPE strain, followed by Enterobacter cloacae and Escherichia coli. A total of 9 carbapenemase types were detected, including Klebsiella pneumoniae carbapenemase (KPC), New Delhi metallo-β-lactamase (NDM), Verona integron- encoded metallo-β-lactamases (VIM), imipenemase (IMP), oxacillinase-48 (OXA-48), and rare imipenem-hydrolyzing β-lactamase (IMI), adelaide imipenemase (AIM), Bicêtre carbapenemase (BIC), and guiana extended-spectrum β-lactamase (GES). The detection rate of KPC serine carbapenemase was 61.7% (309/501). The carbapenemase inhibitor enhancement test exhibited a 100% consistency rate for the strains producing Class A serine carbapenemase and/or Class B metallo-β-lactamases.@*CONCLUSIONS@#CRE strains in Changsha, Hunan, China, are wide distribution and exhibit carbapenemase production. The main mechanism of carbapenem resistance in these bacterias is predominatly attributed to the production of KPC serine carbapenemase. The presence of GES and IMI genes carried by Enterobacterales has been detected for the first time in this region. The carbapenemase inhibitor enhancement test has been proven to be an accurate method for detecting CRE producing Class A serine carbapenemase and/or Class B metallo-β-lactamases. This method offers simpicity of operation and ease of results interpretation, making it weel-suited meeting the clinical microbiology laboratory's reguirements for the detection of serine carbapenemase and metallo-β-lactamases.
Тема - темы
Humans , Carbapenems/pharmacology , Molecular Epidemiology , Bacterial Proteins/analysis , beta-Lactamases/analysis , Klebsiella pneumoniae/genetics , Escherichia coli , Microbial Sensitivity Tests , Serine , Anti-Bacterial Agents/pharmacologyРеферат
Objective To investigate the infection status of infectious diseases for 2 521 patients before blood transfusion in a Hospital in Changsha.Methods A total of 2 521 patients who would be transfused were selected,and six kind of serum hepatitis B virus indicators,hepatitis C virus antibody (antiHCV),human immunodeficiency virus antibody (anti-HIV/1 + 2),and treponema pallidum antibody (antiTP) of nine common infectious disease targets were detected with enzyme-linked immunosorbent assay (ELISA).Results Among 2 521 patients,HBsAg-positive cases were 8.33%,anti-HCV positive were 0.59%,anti-HIV positive [confirmed by the Provincial Center for Disease Control (CDC)] was 6 cases,and TP-positive were 2.301%.A total of 289 patients were tested positively,with a total positive rate of 11.46%.Conclusions Detection before transfusion may reduce infection risk and decrease the risk of occupational exposure,strengthen medical staff self-protection,and reduce medical malpractice caused by blood transfusion.
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Objective To determine diazepam, nitrazepam, oxazepam, estazolam, and al-prazolam simultaneously in human plasma by reversed phase high-performance liquid chromatography (RP-HPLC). Methods Ten microliter carbamazepine (50 mg/L)as the internal standard was added into 1 mL sample, which contained the 5 mixed sedative hypnotics as standard substance and human plasma as ground substance. They were extracted with acetoacetate from plasma samples, and then were dissolved by 100 μL mobile phase. The blood drug levels were analyzed by high perform-ance liquid chromatograph with 20 μL sample injection on a chromatographic column C 18 (4.6 mm×250 mm)at 30℃. The mobile phase consisted of methanol and water (65:35) , and the flow rate was 1.0 mL/min. The ultraviolet detection wavelength was 230 nm. Results The linearity range of the 5 drugs was 5~1 200 μg/L(r≥0.9966, P<0.05). The recovery rate was 95.5%~105.6%. The extraction recovery rate was more than 75%. The relative standard deviation (RSD) of intra-day and inter-day was less than 10% (n=5). Conclusion RP-HPLC method is convert-ient, accurate and sensitive for simultaneous determination of the concentration of diazepam, nitraze-pam, oxazepam, estazolam, and alprazolam in human plasma.
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Objective:With proteomic approach,to find the differentially expressed proteins spots in the peripheral blood mononuclear cells(PBMCs) of healthy people and intracerebral hemorrhage patients with liver yang forming wind syndrome(LYFWS) treated by Zhenganxifeng decoction(ZGXFD) to investigate the possible proteins of LYFWS and the cure mechanism of ZGXFD.Methods: 10 intracerebral hemorrhage patients with LYFWS were treated by ZGXFD for 7-10 days.10 healthy people were selected taken as healthy controls.10ml venous blood was drawed from every subjects and PBMCs were isolated from blood by using lymphocytes separation medium.The total proteins was extracted from PBMCs.The total proteins from either intracerebral hemorrhage patients or healthy controls were prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis.After Coomassie brilliant blue G250 staining,gel-image analysis was performed by PDQuest.The differentially expressed proteins spots were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry(MALD I-TOF-MS).Results: Gel-image analysis revealed that there were 22 proteins spots expressed differentially.20 proteins spots were expressed differentially in the intracerebral hemorrhage patients with LYFWS as compared with healthy control,of which 8 proteins were down-regulated,7 proteins were up-regulated and 5 proteins were lost.After treated by ZGXFD,4 proteins were down-regulated,5 proteins were up-regulated and 2 proteins were lost,9 proteins weren’t expressed di erentially as compared with healthy control.8 of 20 di erentially expressed proteins spots in the intracerebral hemorrhage patients with LYFWS identi ed by MALDI-TOF-MS as compared with healthy control.These proteins are related to cell metabolism,signal transduction and so forth,of which 6 proteins were down-regulated,1 proteins were up-regulated and 1 proteins were lost.After treated by ZGXFD,2 proteins were down-regulated,1 proteins were lost,5 proteins weren’t expressed di erentially as compared with healthy control.Conclusion: 2-DE pro les of intracerebral hemorrhage patients with LYFWS in PBMCs had been established.The proteins of intracerebral hemorrhage patients with LYFWS had been identi ed,which were related to cell metabolism,signal transduction and so forth.This research presented that ZGXFD could adjust multiple di erentially expressed proteins of intracerebral hemorrhage patients with LYFWS.It was also indicated that the cure mechanism of ZGXFD was to regulate multitarget proteins.
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OBJECTIVE To invesigate the prevalence of SHV-producing Enterobacteriaceae isolates in Hunan Province and identify the subtype of SHV encoding gene and the epidemiological aspect of SHV-producing isolates. METHODS Isolates were collected and identified as well as subjected to ESBLs detection.PCR and DNA sequencing were used to determine the genotype of SHV enzymes.The homology of SHV-producing strains were detected by RAPD. RESULTS Twenty-six of 171 Enterobacteriaceae isolates were confirmed to produce blaSHV genes,and 5 subtypes of SHV-type ?-lactamases were determined,including 9 strains of SHV-28,7 strains of SHV-12,7 strains of SHV-1,2 strains of SHV-11 and 1 strain of SHV-5.There were 6 RAPD types in 19 isolates of SHV-producing Klebsiella pneumoniae,5 RAPD types in 5 isolates of SHV-producing Enterobacter cloacae. CONCLUSIONS SHV-12 is the predominant genotype of SHV ESBLs producing Enterobacteriaceae in Hunan Province,and clone spread has played a certain role in SHV-type ?-lactamase producing K.pneumoniae.SHV ESBLs are not found in Escherichia coli.