Prevalence and risk factors for colonisation and infection with carbapenem-resistant Enterobacterales in intensive care units: A prospective multicentre study.

OBJECTIVES: This study aimed to investigate the prevalence and risk factors for carbapenem-resistant Enterobacterales colonisation/infection at admission and acquisition among patients admitted to the intensive care unit. RESEARCH METHODOLOGY/DESIGN: A prospective and multicentre study. SETTING: This study was conducted in 24 intensive care units in Anhui, China. MAIN OUTCOME MEASURES: Demographic and clinical data were collected, and rectal carbapenem-resistant Enterobacterales colonisation was detected by active screening. Multivariate logistic regression models were used to analyse factors associated with colonisation/infection with carbapenem-resistant Enterobacterales at admission and acquisition during the intensive care unit stay. RESULTS: There were 1133 intensive care unit patients included in this study. In total, 5.9% of patients with carbapenem-resistant Enterobacterales colonisation/infection at admission, and of which 56.7% were colonisations. Besides, 8.5% of patients acquired carbapenem-resistant Enterobacterales colonisation/infection during the intensive care stay, and of which 67.6% were colonisations. At admission, transfer from another hospital, admission to an intensive care unit within one year, colonisation/infection/epidemiological link with carbapenem-resistant Enterobacterales within one year, and exposure to any antibiotics within three months were risk factors for colonisation/infection with carbapenem-resistant Enterobacterales. During the intensive care stay, renal disease, an epidemiological link with carbapenem-resistant Enterobacterales, exposure to carbapenems and beta-lactams/beta-lactamase inhibitors, and intensive care stay of three weeks or longer were associated with acquisition. CONCLUSION: The prevalence of colonisation/infection with carbapenem-resistant Enterobacterales in intensive care units is of great concern and should be monitored systematically. Particularly for the 8.5% prevalence of carbapenem-resistant Enterobacterales acquisition during the intensive care stay needs enhanced infection prevention and control measures in these setting. Surveillance of colonisation/infection with carbapenem-resistant Enterobacterales at admission and during the patient's stay represents an early identification tool to prevent further transmission of carbapenem-resistant Enterobacterales. IMPLICATIONS FOR CLINICAL PRACTICE: Carbapenem-resistant Enterobacterales colonization screening at admission and during the patient's stay is an important tool to control carbapenem-resistant Enterobacterales spread in intensive care units.

Identification of Suitable Candidate Reference Genes for Gene Expression Analysis by RT-qPCR in Peripheral Blood Mononuclear Cells of CHB Patients.

BACKGROUND: Quantitative polymerase chain reaction (qPCR) analysis is a precise and effective method for the study of mRNA expression throughout the field of peripheral blood mononuclear cell (PBMC) research. However, the use of suitable reference genes for data normalization is critical to obtain meaningful and reproducible results. The present study aimed to identify the greatest reference genes for further research in PBMC of Chronic Hepatitis B (CHB) patients. METHODS: We assessed the expression stability of four commonly used reference genes (beta actin, beta-tubulin, 18S rRNA, GAPDH) in PBMC of CHB patients. Then we employed geNorm, BestKeeper, and Normfinder to evaluate the expression stability of these reference genes. RESULTS: All four genes displayed no significant differences between patient and control groups except beta actin and thus beta actin should not be used as a normalizing gene in a discussed experimental setup. GAPDH and beta-tubulin composed the best pair of reference genes for normalization purposes in future studies of gene expression in PBMC of CHB patients according to three algorithms. CONCLUSIONS: GAPDH and beta-tubulin were the best combination of two reference genes in this study for RT-qPCR analysis.

The Study of Immune Response in PBMC of CHB Patients treated with IFN-α and 3-TC in vitro.

BACKGROUND: The primary aim of this study is to measure the JAK-STAT signaling in HBV infected peripheral blood mononuclear cells (PBMCs) stimulated by IFN-α and 3-TC and explore the influence of HBV to the JAKSTAT signaling pathways. METHODS: PBMCs were separated from healthy volunteers and patients who had not received any treatment with chronic hepatitis B. PBMCs were divided into the control group, IFN-α stimulation group, Lamivudine stimulation group, and combined treatment group. The expression of molecules of JAK-STAT signal transduction pathway (STAT1, STAT2, IRF9) and the antiviral protein (MxA) were detected by RT-qPCR and Western blot method. RESULTS: The majority of IFN-α inducible genes were expressed. The molecules of JAK-STAT signal transduction pathway (STAT1, STAT2, IRF9) and the antiviral protein (MxA) were highly expressed in IFN-α stimulation group and the combined treatment group. Compared to healthy controls, the expression levels of molecules (STAT1, IRF9) and the antiviral protein (MxA) are significantly lower in the control group, IFN-α stimulation group, and the combined treatment group of the CHB patients. CONCLUSIONS: IFN-α could activate JAK-STAT signaling transduction pathway in PBMCs of HBV-infected patients and HBV might process the activity to antagonize the antiviral activity in HBV infected PBMCs.