Continuous glucose monitoring profile in COVID-19 patients with and without diabetes receiving methylprednisolone.

PURPOSE: Methylprednisolone is widely used during the COVID-19 epidemic. We aimed to evaluate the glucose profile of COVID-19 patients with and without diabetes receiving methylprednisolone. METHODS: 36 patients with COVID-19 admitted to hospital were included: 17 with and 19 without diabetes. Methylprednisolone 40 mg was administered at about 9:00 a.m. Glucose levels were assessed by blinded intermittently scanned continuous glucose monitoring (isCGM) for an average of 6.8 ± 2.4 days. Excess hyperglycemia was defined as time above range (TAR) > 10.0 mmol/L (TAR>10.0) ≥ 25%, or TAR > 13.9 mmol/L (TAR>13.9) ≥ 10%. RESULTS: Glucose management indicator (GMI) was significantly higher than the admission glycated hemoglobin A1c (HbA1c) level in patients without diabetes [6.7 (6.1-7.0) % vs. 5.9 (5.9-6.1) %, P < 0.001], while no significant difference was found in patients with diabetes [9.0 (7.5-9.5) % vs. 8.9 (7.5-10.2) %, P > 0.05]. The difference between GMI and HbA1c (∆GMI-HbA1c) in patients without diabetes was significantly higher than in patients with diabetes [0.7 (0.2-1.0) % vs. -0.2 (-1.5-0.5) %, P = 0.005]. The circadian patterns of glucose were similar in the two groups. In patients without diabetes, excess hyperglycemia occurred in 31.6% (6/19) of participants, with 31.6% (6/19) having a TAR>10.0 ≥ 25%, while 21.1% (4/19) had a TAR>13.9 ≥ 10%. CONCLUSION: The impact of methylprednisolone on glycemia was more pronounced in COVID-19 patients without diabetes, compared to those with diabetes. A significant burden of methylprednisolone-induced hyperglycemia was observed in patients without diabetes.

USP9x promotes CD8 + T-cell dysfunction in association with autophagy inhibition in septic liver injury.

Sepsis is a life-threatening condition manifested by concurrent inflammation and immunosuppression. Ubiquitin-specific peptidase 9, X-linked (USP9x), is a USP domain-containing deubiquitinase which is required in T-cell development. In the present study, we investigate whether USP9x plays a role in hepatic CD8 + T-cell dysfunction in septic mice. We find that CD8 + T cells are decreased in the blood of septic patients with liver injury compared with those without liver injury, the CD4/CD8 ratio is increased, and the levels of cytolytic factors, granzyme B and perforin are downregulated. The number of hepatic CD8 + T cells and USP9x expression are both increased 24 h after cecal ligation and puncture-induced sepsis in a mouse model, a pattern similar to liver injury. The mechanism involves promotion of CD8 + T-cell dysfunction by USP9x associated with suppression of cell cytolytic activity via autophagy inhibition, which is reversed by the USP9x inhibitor WP1130. In the in vivo studies, autophagy is significantly increased in hepatic CD8 + T cells of septic mice with conditional knockout of mammalian target of rapamycin. This study shows that USP9x has the potential to be used as a therapeutic target in septic liver injury.

Characterization of lncRNA-Based ceRNA Network and Potential Prognostic Hub Genes for Sepsis.

Objective: Sepsis is one of the most common reasons for hospitalization and in-hospital mortality each year. Noncoding RNAs have been reported not only as diagnostic and prognostic indicators but also as therapeutic targets of sepsis. Herein, we used an integrative computational approach to identify miRNA-mediated ceRNA crosstalk between lncRNAs and genes in sepsis based on the "ceRNA hypothesis" and investigated prognostic roles of hub genes in sepsis. Methods: Two good-quality gene expression datasets with more than 10 patient samples, GSE89376 and GSE95233, were employed to obtain differentially expressed lncRNAs (DElncRNAs) and genes (DEGs) in sepsis. The DElncRNA-miRNA-DEG regulatory network was constructed using a combination of DElncRNA-miRNA pairs and miRNA-DEmRNA pairs. The protein-protein interaction (PPI) network was constructed by mapping DEGs into the STRING database to identify hub genes in sepsis. The clinical and prognostic significance of hub genes was validated in 89 patients with post-traumatic sepsis. Results: The integrative computational approach identified 311 DEGs and 19 DElncRNAs between septic patients and healthy volunteers. Results yielded 122 downDElncRNA-miRNA-downDEG networks based on two lncRNAs, HCP5, and HOTAIRM1, and 36 upDElncRNA-miRNA-upDEG network based on BASP1-AS1. The PPI network identified serum/glucocorticoid regulated kinase 1 (SGK1), arrestin beta 1 (ARRB1), and G protein-coupled receptor 183 (GPR183) as located at the core of the network, and three of them were downregulated in sepsis. SGK1, ARRB1, and GPR183 were all involved in lncRNA HCP5-based ceRNA network. The quantitative real-time PCR revealed that the patients with post-traumatic sepsis exhibited reduced relative mRNA levels of SGK1, ARRB1, and GPR183 compared to the patients without sepsis. The nonsurvivor group, according to the 28-day mortality, showed lower relative mRNA levels of SGK1, ARRB1, and GPR183 than the survivor group. We also demonstrated reduced mRNA levels of SGK1, ARRB1, and GPR183 were associated with sepsis-related death after trauma. Conclusion: Our integrative analysis and clinical validation suggest lncRNA HCP5-based ceRNA networks with SGK1, ARRB1, and GPR183 involved were associated with the occurrence and progression of sepsis.

Risk Factors and Outcome of Sepsis in Traumatic Patients and Pathogen Detection Using Metagenomic Next-Generation Sequencing.

Objective: Sepsis, a life-threatening clinical syndrome, is a leading cause of mortality after experiencing multiple traumas. Once diagnosed with sepsis, patients should be given an appropriate empiric antimicrobial treatment followed by the specific antibiotic therapy based on blood culture due to its rapid progression to tissue damage and organ failure. In this study, we aimed to analyze the risk factors and outcome of sepsis in traumatic patients and to investigate the performance of metagenomic next-generation sequencing (mNGS) compared with standard microbiological diagnostics in post-traumatic sepsis. Methods: The study included 528 patients with multiple traumas among which there were 142 cases with post-traumatic sepsis. Patients' demographic and clinical data were recorded. The outcome measures included mortality during the emergency intensive care unit (EICU), EICU length of stay (LOS), all-cause 28-day mortality, and total ventilator days in 28 days after admission. A total of 89 blood samples from 89 septic patients underwent standard microbiological blood cultures and 89 samples of peripheral blood (n = 21), wound secretion (n = 41), bronchoalveolar lavage fluid (BALF) (19), ascites (n = 5), and sputum (n = 3) underwent mNGS. Pathogen detection was compared between standard microbiological blood cultures and mNGS. Results: The sepsis group and non-sepsis group exhibited significant differences regarding shock on admission, blood transfusion, mechanical ventilation, body temperature, heart rate, WBC count, neutrophil count, hematocrit, urea nitrogen, creatinine, CRP, D-D dimer, PCT, scores of APACHE II, sequential organ failure assessment (SOFA), and Injury Severity Score (ISS) on admission to the EICU, and Multiple Organ Dysfunction Syndromes (MODS) (P < 0.05). Multivariate logistic regression analysis showed that scores of APACHE II, SOFA, and ISS on admission, and MODS were independent risk factors for the occurrence of sepsis in patients with multiple traumas. The 28-day mortality was higher in the sepsis group than in the non-sepsis group (45.07% vs. 19.17%, P < 0.001). The mortality during the EICU was higher in the sepsis group than in the non-sepsis group (P=0.002). The LOS in the EICU in the sepsis group was increased compared with the non-sepsis group (P=0.004). The total ventilator days in 28 days after admission in the sepsis group was increased compared with the non-sepsis group (P < 0.001). Multivariate logistic regression analysis showed that septic shock, APACHE II score on admission, SOFA score, and MODS were independent risk factors of death for patients with post-traumatic sepsis. The positive detection rate of mNGS was 91.01% (81/89), which was significantly higher than that of standard microbiological blood cultures (39.33% (35/89)). Standard microbiological blood cultures and mNGS methods demonstrated double positive results in 33 (37.08%) specimens and double-negative results in 8 (8.99%) specimens, while 46 (51.69%) samples and 2 (2.25%) samples had positive results only with mNGS or culture alone, respectively. Conclusion: Our study identifies risk factors for the incidence and death of sepsis in traumatic patients and shows that mNGS may serve as a better diagnostic tool for the identification of pathogens in post-traumatic sepsis than standard microbiological blood cultures.

Identification of Hub Genes With Differential Correlations in Sepsis.

As a multifaceted syndrome, sepsis leads to high risk of death worldwide. It is difficult to be intervened due to insufficient biomarkers and potential targets. The reason is that regulatory mechanisms during sepsis are poorly understood. In this study, expression profiles of sepsis from GSE134347 were integrated to construct gene interaction network through weighted gene co-expression network analysis (WGCNA). R package DiffCorr was utilized to evaluate differential correlations and identify significant differences between sepsis and healthy tissues. As a result, twenty-six modules were detected in the network, among which blue and darkred modules exhibited the most significant associations with sepsis. Finally, we identified some novel genes with opposite correlations including ZNF366, ZMYND11, SVIP and UBE2H. Further biological analysis revealed their promising roles in sepsis management. Hence, differential correlations-based algorithm was firstly established for the discovery of appealing regulators in sepsis.

MiR-29b-3p Inhibits the Inflammation Injury in Human Umbilical Vein Endothelial Cells by Regulating SEC23A.

This study aims to investigate the effects of miR-29b-3p on the inflammation injury of human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS) and explore the underlying mechanisms. The effects of different concentrations of LPS (0, 1, 5 and 10 µg/mL) on inflammation injury in HUVECs are detected by ELISA, CCK-8, EdU, flow cytometry and western blot analyses to determine the optimal stimulus concentration. After stimulating HUVECs with 10 µg/mL LPS, the expression levels of miR-29b-3p are detected, and the effects of miR-29b-3p on inflammation injury are detected by ELISA, CCK-8, EdU, flow cytometry and western blot analyses. Bioinformatic analysis, luciferase reporter assay and confirmatory experiments are applied to identify the target gene bound with miR-29b-3p. Rescue experiments have verified the roles of miR-29b-3p and the target gene in inflammation injury. We found that pro-inflammatory factor was increased, apoptosis was promoted, and cell proliferation was inhibited after the treatment of LPS in HUVECs. Overexpression of miR-29b-3p inhibited LPS-induced inflammatory response and apoptosis while promoting proliferation in HUVECs. Besides, bioinformatics analysis indicated that SEC23A was the target gene of miR-29b-3p and the confirmatory experiments showed that SEC23A was negatively correlated with miR-29b-3p and positively correlated with LPS concentration. Rescue experiments revealed that overexpression of SEC23A partially enhanced the inflammation injury effects in LPS-induced HUVECs with overexpression of miR-29b-3p. Hence, miR-29b-3p repressed inflammatory response, cell apoptosis and promoted cell proliferation in LPS-induced HUVECs by targeting SEC23A, providing a potential target for treating sepsis.

Data-independent acquisition-based quantitative proteomic analysis reveals differences in host immune response of peripheral blood mononuclear cells to sepsis.

This study was aimed to uncover proteins that are differentially expressed in sepsis. Data-independent acquisition (DIA) was used for analysis to identify differentially expressed proteins in peripheral blood mononuclear cells (PBMCs) of patients. A total of 24 non-septic intensive care unit (ICU) patients, 11 septic shock patients and 27 patients diagnosed with sepsis were recruited for the mass spectrometry (MS) discovery. PBMCs were isolated from routine blood samples and digested into peptides. A DIA workflow was developed using a quadrupole-Orbitrap liquid chromatography LC-MS system, and mass spectra peaks were extracted by Skyline software. Orthogonal partial least-squares discriminant analysis (OPLS-DA) and partial least-squares discriminant analysis (PLS-DA) were applied to distinguish the patient groups at the level of fragment ion and peptide. Differentially expressed proteins in the patient groups were verified by enzyme-linked immunosorbent assay (ELISA). Receiver-operating characteristic (ROC) curves were used to evaluate the protein expression. A total of 1062 fragment ions and 122 proteins were identified in the MS-DIA analysis conducted by Skyline software. Using gene ontology clustering analysis, we discovered that 51 of the 122 identified proteins were associated with biological processes, including carbon metabolism, biosynthesis of antibiotics, platelet activation, bacterial invasion of epithelial cells and complement, and coagulation cascades. Among them, five proteins (high-mobility group box1 [HMGB1], matrix metalloproteinase 8 [MMP8], neutrophil gelatinase-associated lipocalin [NGAL], lactotransferrin [LTF] and grancalcin [GCA]) were identified by ELISA as closely related to the development of sepsis. The ROC curves displayed good sensitivity and specificity.

Candida albicans infection and intestinal immunity.

Fungal infections cause high rates of morbidity and mortality in intensive care and immunocompromised patients, and can represent a life-threatening disease. As a microorganism commonly found in the intestine, Candida albicans (C. albicans) can invade the gut epithelium barrier via microfold cells and enter the bloodstream. The defensive potential of the intestinal barrier against invasive C. albicans is dependent on innate and adaptive immune responses which enable the host to eliminate pathogenic fungi. The lamina propria layer of the intestine contains numerous immune cells capable of inducing an innate cellular immune response against invasive fungi. This review focuses on the immune response triggered by a C. albicans infection in the intestine.