ABSTRACT
Objective:To explore whether the lipopolysaccharide (LPS)-induced modification of O-linked N-acetylglucosamine (O-GlcNAc) is involved in the inflammatory signaling pathway of endothelial cells.Methods:Human umbilical vein endothelial cells (HUVEC) were cultured in vitro, and cells in logarithmic growth phase were used for experiments. Cells were divided into blank control group, LPS group (2 000 mg/L LPS), O-GlcNAc transferase (OGT) overexpression (OGT-OE)+LPS group (plasmid transfection OGT+2 000 mg/L LPS), protein kinase C (PKC) inhibitor+LPS group (10 μmol/L Go 6983+2 000 mg/L LPS), RhoA inhibitor+LPS group (40 μmol/L Rhoin hydrochloride+2 000 mg/L LPS), phosphatidylinositol-3-kinase (PI3K) inhibitor+LPS group (1 μmol/L SL-2052+2 000 mg/L LPS), serine/threonine kinase (Akt) inhibitor+LPS group (10 μmol/L PP2+2 000 mg/L LPS) and small interfering RNA (siRNA) treated Akt (si-AKT)+LPS group (si-Akt+2 000 mg/L LPS). After 24 hours of LPS treatment, real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect the transcription levels of inflammatory cytokines [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1)]. The protein expression or phosphorylation of OGT, O-GlcNAc, Akt, extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinase (p38MAPK), nuclear factor-κB p65 (NF-κB p65), and signal transducer and activator of transcription 3 (STAT3) were determined by Western blotting. Results:Compared with the blank control group, the expression of OGT and the modification of O-GlcNAc in the LPS group were decreased, while the expressions of phosphorylated ERK, p38MAPK, and STAT3 were increased, and the transcript levels of inflammatory cytokines were also significantly increased [IL-6 mRNA (2 -ΔΔCt): 4.71±0.60 vs. 1.03±0.29, TNF-α mRNA (2 -ΔΔCt): 1.89±0.11 vs. 1.04±0.35, ICAM-1 mRNA (2 -ΔΔCt): 2.06±0.18 vs. 1.02±0.21, VCAM-1 mRNA (2 -ΔΔCt): 2.94±0.57 vs. 1.01±0.17, all P < 0.05], indicating that LPS could decrease O-GlcNAc modification, activate inflammatory signaling pathways and increase inflammatory cytokines expression. Compared with the LPS group, the expressions of phosphorylated ERK, p38MAPK, NF-κB p65, and STAT3 in the endothelial cells of the OGT-OE+LPS group were decreased, and the expression of inflammatory factors were significantly decreased [IL-6 mRNA (2 -ΔΔCt): 0.12±0.01 vs. 0.90±0.17, TNF-α mRNA (2 -ΔΔCt): 0.31±0.01 vs. 0.91±0.14, ICAM-1 mRNA (2 -ΔΔCt): 0.64±0.02 vs. 1.13±0.16, VCAM-1 mRNA (2 -ΔΔCt): 0.11±0.01 vs. 0.93±0.11, all P < 0.05], indicating that the increase of OGT level could inhibit the partial activation of the endothelial inflammatory signal pathway under the LPS stimulation. Compared with the blank control group, the phosphorylation level of Akt in the LPS group was increased. Compared with the LPS group, both OGT expression and O-GlcNAc modification were down-regulated after pretreatment of PKC inhibitor, RhoA inhibitor, PI3K inhibitor, or Akt inhibitor. Compared with the LPS group, the transcript levels of IL-6, TNF-α and ICAM-1 in the PP2+LPS group were significantly decreased [IL-6 mRNA (2 -ΔΔCt): 1.46±0.16 vs. 3.55±0.87, TNF-α mRNA (2 -ΔΔCt): 0.98±0.14 vs. 1.76±0.10, ICAM-1 mRNA (2 -ΔΔCt): 1.39±0.24 vs. 2.04±0.13, all P < 0.05], but there was no significant change in VCAM-1. Compared with the LPS group, the expression of OGT and O-GlcNAc modification in the si-Akt+LPS group were decreased, while the transcript levels of inflammatory cytokines were also significantly decreased [IL-6 mRNA (2 -ΔΔCt): 0.75±0.03 vs. 0.99±0.09, TNF-α mRNA (2 -ΔΔCt): 0.69±0.01 vs. 1.10±0.08, ICAM-1 mRNA (2 -ΔΔCt): 0.76±0.01 vs. 0.99±0.02, VCAM-1 mRNA (2 -ΔΔCt): 0.93±0.08 vs. 1.20±0.21, all P < 0.05], indicating that Akt participated in the action process of LPS on OGT and affected the inflammatory factor expression. Conclusions:The decreased level of O-GlcNAc modification in endothelial cells stimulated with LPS promotes partial activation of inflammatory signaling pathways, mainly involving ERK, p38MAPK, and STAT3, and affects the expression of inflammatory factors. AKT may be involved in the effect of LPS on the inhibition of O-GlcNAc modification.
ABSTRACT
Objective:To compare the characteristics of patients undergoing blood purification treatment in PICU of a children′s tertiary hospital during 8 years, so as to analyze the changes in the development of blood purification technology in children in East China.Methods:Patients who received blood purification treatment in PICU of Children′s Hospital of Fudan University from 2014 to 2021 were included and divided into two study periods: 2014-2017 and 2018-2021.The clinical characteristics and treatment parameters of patients were collected and analyzed.Results:A total of 1 029 patients were included in the study, of which 103 were combined with extracorporeal membrane oxygenation.The 28-day survival rate of 926 patients treated with pure blood purification was 55.7%.Among them, patients with younger age, lower body weight, using mechanical ventilation, using vasoactive drugs before blood purification, and patients with multiple organ dysfunction syndrome had a higher distribution in the death group than those in survival group( P<0.05). During 8 years, a total of 3 688 cases of blood purification were performed.The main mode was continuous veno-venous hemodiafiltration (CVVHDF) (68.6%), followed by therapeutic plasma exchange (TPE) (23.8%) and hemoperfusion (HP) (4.8%); the main indication was acute kidney injury (AKI) (29.3%), followed by severe inflammatory disease (26.2%) and acute liver failure (16.2%). Compared with 2014-2017, the number of blood purification treatments in 2018-2021 increased by 47.4%, and the survival rate of patients increased significantly (48.7% vs. 58.1%, P<0.05). The distribution of blood purification patterns and indications also changed( P<0.05). The proportions of TPE (20.5% vs. 26.0%) and HP (3.1% vs. 6.0%) increased, while the proportion of CVVHDF (71.9% vs. 66.4%) decreased significantly.The proportions of AKI (29.8% vs. 38.9%) and refractory immune diseases (8.4% vs. 15.2%) were significantly higher, while severe inflammatory diseases (29.2% vs. 24.2%) and acute liver failure (19.6% vs. 13.8%) had declined. Conclusion:From 2014 to 2021, the number of blood purifications performed in our center increased significantly.Although the distribution of indications and patterns have also changed significantly, the overall survival rate is significantly improved.However, standardized practice still needs to be strengthened.
ABSTRACT
Adenovirus pneumonia is a common type of pneumonia in immunocompetent and immunocompromised patients, and its prognosis is poor.Antiviral therapy includs ganciclovir, valganciclovir, cidofovir, brincidofovir, ribavirin and so on.Among them, cidofovir and brincidofovir have obvious antiviral activity against adenovirus in vitro and in vivo, but further RCT results are still needed.Therefore, antiviral therapy of adenovirus pneumonia still needs further study.
ABSTRACT
Adenovirus pneumonia is a common type of pneumonia in immunocompetent and immu-nocompromised patients,and its prognosis is poor. Antiviral therapy includs ganciclovir,valganciclovir,cido-fovir,brincidofovir,ribavirin and so on. Among them,cidofovir and brincidofovir have obvious antiviral activ-ity against adenovirus in vitro and in vivo,but further RCT results are still needed. Therefore,antiviral therapy of adenovirus pneumonia still needs further study.
ABSTRACT
The mortality of sepsis remains high,and it is still a major cause of death in critically ill patients.But its pathophysiological mechanism remains a mystery,not yet fully clear.Endothelium play an important role in the development of sepsis.Maintaining the integrity of its surface glycocalyx to a certain extent reduces sepsis damage;and its destruction mediates a series of pathophysiological responses,including increasing vascular permeability by cell pathway or intracellular pathways,increasing the adhesion of white blood cells to endothelial cejls and a variety of cytokines to promote inflammatory response,activating of thrombin to coagulation dysfunction,promoting synthesis of nitric oxide to change vascular tension.The damage of glycocalyx as a first step in endothelial cell injury can be used as early diagnosis and prognosis of sepsis,which provides new ideas for the diagnosis and treatment of sepsis.