LncRNA CASC2 Alleviates Sepsis-induced Acute Lung Injury by Regulating the miR-152-3p/PDK4 Axis.

BACKGROUND: Acute lung injury (ALI) is an early complication of sepsis and it is also considered as an important cause of high mortality in sepsis patients. This research aimed to explore the potential role and mechanism of long non-coding RNA (lncRNA) cancer susceptibility candidate 2 (CASC2) in sepsis-induced ALI. METHODS: The levels of CASC2, microRNA-152-3p (miR-152-3p) and pyruvate dehydrogenase kinase 4 (PDK4) in sepsis patients and LPS-treated HPAEpiC were detected by quantitative real-time PCR and western blot. Cell viability and apoptosis were assessed by Counting Kit-8 (CCK-8) assay and flow cytometry. The concentrations of inflammatory factors were tested by Enzyme-linked immunosorbent assay. Oxidative stress was evaluated by the levels of reactive oxygen species and superoxide dismutase using corresponding commercial kits. The targeting relationship between miR-152-3p and CASC2 or PDK4 was verified by dual-luciferase reporter, RNA immunoprecipitation (RIP) and RNA pull-down assays. RESULTS: CASC2 and PDK4 were down-regulated, while miR-152-3p was up-regulated in sepsis patients and LPS-stimulated HPAEpiC. Overexpression of CASC2 relieved the LPS-resulted cell viability inhibition, apoptosis promotion, inflammatory and oxidative damages in HPAEpiC. In addition, miR-152-3p was a miRNA target of CASC2 and CASC2 alleviated cell injury in LPS-disposed HPAEpiC by sponging miR-152-3p. Moreover, miR-152-3p directly targeted PDK4 and CASC2 increased the PDK4 expression by depending on the sponge effect on miR-152-3p. Meanwhile, inhibition of miR-152-3p attenuated LPS-triggered HPAEpiC injury by upregulating the level of PDK4. CONCLUSION: These results suggested that CASC2 ameliorated the LPS-induced injury in HPAEpiC via regulating miR-152-3p/PDK4 pathway.

Actively implementing an evidence-based feeding guideline for critically ill patients (NEED): a multicenter, cluster-randomized, controlled trial.

BACKGROUND: Previous cluster-randomized controlled trials evaluating the impact of implementing evidence-based guidelines for nutrition therapy in critical illness do not consistently demonstrate patient benefits. A large-scale, sufficiently powered study is therefore warranted to ascertain the effects of guideline implementation on patient-centered outcomes. METHODS: We conducted a multicenter, cluster-randomized, parallel-controlled trial in intensive care units (ICUs) across China. We developed an evidence-based feeding guideline. ICUs randomly allocated to the guideline group formed a local "intervention team", which actively implemented the guideline using standardized educational materials, a graphical feeding protocol, and live online education outreach meetings conducted by members of the study management committee. ICUs assigned to the control group remained unaware of the guideline content. All ICUs enrolled patients who were expected to stay in the ICU longer than seven days. The primary outcome was all-cause mortality within 28 days of enrollment. RESULTS: Forty-eight ICUs were randomized to the guideline group and 49 to the control group. From March 2018 to July 2019, the guideline ICUs enrolled 1399 patients, and the control ICUs enrolled 1373 patients. Implementation of the guideline resulted in significantly earlier EN initiation (1.20 vs. 1.55 mean days to initiation of EN; difference - 0.40 [95% CI - 0.71 to - 0.09]; P = 0.01) and delayed PN initiation (1.29 vs. 0.80 mean days to start of PN; difference 1.06 [95% CI 0.44 to 1.67]; P = 0.001). There was no significant difference in 28-day mortality (14.2% vs. 15.2%; difference - 1.6% [95% CI - 4.3% to 1.2%]; P = 0.42) between groups. CONCLUSIONS: In this large-scale, multicenter trial, active implementation of an evidence-based feeding guideline reduced the time to commencement of EN and overall PN use but did not translate to a reduction in mortality from critical illness. TRIAL REGISTRATION: ISRCTN, ISRCTN12233792 . Registered November 20th, 2017.

Circular RNA circVMA21 ameliorates lipopolysaccharide (LPS)-induced acute kidney injury by targeting the miR-199a-5p/NRP1 axis in sepsis.

BACKGROUND: Sepsis is a serious and elusive syndrome caused by infection, with high mortality worldwide. Circular RNAs vacuolar ATPase assembly factor (circVMA21) has been reported to be related to the inflammatory damages in sepsis. This study is designed to explore the role and mechanism of circVMA21 in the lipopolysaccharide (LPS)-induced cell injury in sepsis. METHODS: Cell viability and apoptosis were detected by CCK-8, and flow cytometry assays. CircVMA21, microRNA-199a-5p (miR-199a-5p), and Neuropilin-1 (NRP1) level were determined by RT-qPCR. Protein levels of Bcl-2, Bax, cleaved-caspase 3, and NRP1 were examined by Western blot assay. IL-1ß, IL-6, and TNF-α were detected using ELISA. Superoxide Dismutase (SOD) and glutathione (GSH) were measured by the special kits. The binding relationship between miR-199a-5p and circVMA21 or NRP1 was predicted by Starbase 3.0 and then verified by a dual-luciferase reporter and RIP assays. RESULTS: CircVMA21 and NRP1 were decreased, and miR-199a-5p was increased in LPS-induced THP-1 cells. Moreover, circVMA21 overexpression could repress LPS-mediated cell viability, apoptosis, inflammation, and oxidative stress in THP-1 cells. The mechanical analysis suggested that circVMA21 regulated NRP1 expression through sponging miR-199a-5p. CONCLUSION: CircVMA21 upregulation could attenuate LPS-triggered THP-1 cell injury through modulating the miR-199a-5p/NRP1 axis, hinting an underlying therapeutic strategy for sepsis patients.

Circ_RASGEF1B Promotes LPS-Induced Apoptosis and Inflammatory Response by Targeting MicroRNA-146a-5p/Pdk1 Axis in Septic Acute Kidney Injury Cell Model.

BACKGROUND: We intended to investigate the function of circular RNA RasGEF domain family member 1B (circ_RASGEF1B) in lipopolysaccharide (LPS)-induced septic acute kidney injury (AKI) cell model and its associated mechanism. METHODS: TCMK-1 cells were exposed to 10 µg/mL LPS for 24 h to establish a septic AKI cell model. Mice were intraperitoneally injected with 10 mg/kg LPS to establish a septic AKI mice model. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot assay were used to measure RNA and protein expression, respectively. Cell viability and apoptosis were assessed by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry. Cell inflammatory response was analyzed using enzyme-linked immunosorbent assay. Dual-luciferase reporter assay was conducted to confirm the predicted target relationship between microRNA-146a-5p (miR-146a-5p) and circ_RASGEF1B or pyruvate dehydrogenase kinase 1 (Pdk1). RESULTS: The circ_RASGEF1B level was upregulated in LPS-induced TCMK-1 cells and septic AKI mice models. LPS exposure reduced cell viability and promoted cell apoptosis and inflammatory response partly by upregulating circ_RASGEF1B. Circ_RASGEF1B bound to miR-146a-5p and miR-146a-5p interference partly overturned circ_RASGEF1B silencing-mediated effects in LPS-induced TCMK-1 cells. Pdk1 was a target of miR-146a-5p, and Pdk1 accumulation partly counteracted miR-146a-5p-induced influences in TCMK-1 cells upon LPS stimulation. CONCLUSION: Circ_RASGEF1B promoted LPS-induced apoptosis and inflammatory response in renal tubular epithelial cells partly by upregulating Pdk1 via acting as miR-146a-5p sponge.

The impact of transthoracic echocardiography on the short-term prognosis of elderly patients in the intensive care unit: a retrospective analysis based on the MIMIC-III database.

BACKGROUND: Elderly patients in the intensive care unit (ICU) often suffer from cardiac function impairment. Real-time monitoring of cardiac function and structural changes has important clinical significance. Transthoracic echocardiography (TTE) is a convenient, intuitive, and non-invasive real-time examination of the heart, and it has been widely used for intensive care patients. This study aims to analyze the impact of TTE on the prognosis of elderly patients in ICU. METHODS: Data from elderly patients in the ICU was obtained from the MIMIC-III 1.4 database, and they were divided into a TTE examination group and a non-TTE examination group. The baseline data of the two groups were compared, and multiple regression analysis, propensity score (PS), compatibility analysis, and other methods were used to analyze the influence of TTE on the prognosis of elderly patients in ICU. RESULTS: A total of 8,952 elderly cases were included, comprising 3,280 cases (36.6%) in the TTE group and 5,672 cases (63.4%) in the non-TTE group. The SAPS score (20.34±5.34 vs. 18.74±5.2, t=13.889, P<0.001) and SOFA score (5.10±3.38 vs. 3.82±2.81, t=19.250, P<0.001) of patients in the TTE group were higher than those of non-TTE group. The rate of patients in the TTE group receiving mechanical ventilation (52.10% vs. 34.80%) and vasoactive drugs (29.30% vs. 15.00%) was significantly higher than that in the non-TTE group. In the PS score compatibility cohort, the 28-day mortality rate of patients in the TTE group was 23.4%, and the 28-day mortality rate of patients in the non-TTE group was 28.7%. The adjusted odd ratio (OR) value was 0.76 (95% CI: 0.65-0.87, P<0.001). Analysis of secondary endpoints showed that patients in the TTE group did not use mechanical ventilation and hypertension drugs for a longer period of time than those in the non-TTE group, and the TTE group patients had significantly more fluid input in the first three days after admission to the ICU than in the non-TTE group. CONCLUSIONS: TTE examination can reduce the 28-day mortality risk of elderly critically ill patients.

Analysis of prediction and early warning indexes of patients with COVID-19.

BACKGROUND: The aim was to compare the laboratory data of patients with suspected and confirmed new coronavirus pneumonia (COVID-19) and look for diagnostic predictive and early warning indicators, which will help to better manage the disease. METHODS: A total of 36 confirmed COVID-19 patients were divided into the general (n = 17) and critical group (n = 19). The suspected group enrolled 23 suspected COVID-19 patients with the negative nucleic acid test result. We collected all patients' clinical characteristics and some laboratory indicators at the time of admission and conducted Logistic regression analysis after comparing the differences between groups. RESULTS: There were no significant differences in age, gender, disease duration, fever history, and comorbidities between the suspected and general group (P > 0.05); however, fibrinogen was statistically different (P < 0.05). Compared with the general group, the oxygenation index and lymphocytes were significantly reduced and the Neutrophil-to-lymphocyte Ratio (NLR) and total bilirubin were increased in the critical group (P < 0.05). The fibrinogen OR value was 2.52 (95% CI 1.18-5.36, P = 0.017) and the NLR OR value was 2.91 (95% CI 1.36-6.21, P = 0.006). CONCLUSIONS: Fibrinogen is a valuable diagnostic predictor for patients with suspected COVID-19. For confirmed COVID-19 patients, the NLR is a valuable early warning indicator.

[Early diagnostic value of neutrophil gelatinase-associated lipocalin and interleukin-18 in patients with sepsis-induced acute kidney injury].

OBJECTIVE: To evaluate the early diagnosis value of blood neutrophil gelatinase-associated lipocalin (NGAL) and interleukin-18 (IL-18) in patients with sepsis-induced acute kidney injury (AKI). METHODS: A prospective case controlled study was conducted. Fifty-six patients with sepsis but without renal disease admitted to intensive care unit (ICU) of Shanxi People's Hospital from April 2014 to April 2015 were enrolled. Blood lactic acid and acute physiology and chronic health evaluation II (APACHE II) score at ICU admission were recorded. The urine output, blood urea nitrogen (BUN), serum creatinine (SCr), NGAL, IL-18 and tumor necrosis factor-α (TNF-α) were continuously monitored for 48 hours. The patients were divided into AKI group and non-AKI group according to the diagnostic criteria of Kidney Disease: Improving Global Outcomes (KDIGO). The changes trend in above parameters between the two groups at different time points were compared. The early diagnostic value of NGAL, IL-18, and SCr for sepsis-induced AKI was evaluated by receiver operating characteristic curve (ROC). RESULTS: Compared with non-AKI group, with the time prolongation after ICU admission, the traditional parameters reflecting AKI urine output in sepsis-induced AKI group was decreased gradually, BUN and SCr were gradually increased, and a statistically significant difference was found at 12 hours between the two groups [BUN (mmol/L): 11.8±3.5 vs. 8.2±3.5, SCr (µmol/L): 88.6±11.3 vs. 74.0±11.0, both P < 0.01]; but the new indicators NGAL and IL-18 reflecting AKI had a statistically significant difference at 6 hours [NGAL (µg/L): 426.7±90.7 vs. 382.3±67.9, IL-18 (ng/L): 75.7±9.3 vs. 70.9±7.3, both P < 0.05]. It was shown that in AKI group, NGAL and IL-18 compared with BUN and SCr increased at least 6 hours ahead of schedule. The area under ROC curve (AUC) of 6-hour NGAL and IL-18 in patients with sepsis-induced AKI were 0.821 [95% confidence interval (95%CI) = 0.713-0.931] and 0.719 (95%CI = 0.584-0.853) respectively, superior to SCr (AUC = 0.677, 95%CI = 0.528-0.825). The cutoff value of NGAL was 363.58 µg/L, and the sensitivity and specificity were 88.0% and 86.7% respectively. CONCLUSIONS: NGAL and IL-18 in the early prediction of sepsis patients with AKI are better than SCr, and NGAL was most sensitive. Therefore, NGAL can be used as an early biomarker for the diagnosis of AKI in patients with sepsis.