To investigate the effect of granulocyte colony-stimulating factor receptor (G-CSFR) on the expression of syndecan-4 (SDC4) in endothelial cells during sepsis-associated acute lung injury (ALI) and its regulatory mechanisms.
Methods
A total of 30 C57bl/6 mice were randomly divided into three groups using a random number table: a negative control group (sham group, intraperitoneal injection of an isotonic NaCl solution of equal concentration), a sepsis-associated ALI group [lipopolysaccharide (LPS) group, intraperitoneal injection of 20 mg/kg LPS], and a LPS combined with short hairpin RNA targeting G-CSFR (shG-CSFR) intervention group [LPS + shG-CSFR group, intratracheal injection of shG-CSFR followed by intraperitoneal injection of LPS], with 10 mice in each group. Hematoxylin-eosin staining, immunofluorescence staining and real-time quantitative PCR were used to detect the expression levels of inflammatory factors, the degree of inflammatory damage, and the fluorescence intensities of G-CSFR and SDC4 in lung tissue. For in vitro experiments, pulmonary microvascular endothelial cells were subjected to LPS stimulation, lentivirus overexpression, and PP2 [a non-receptor tyrosine kinase (SRC) inhibitor] treatment. RNA sequencing (RNA-seq), real-time quantitative PCR, and western-blotting were used to identify differentially expressed genes following G-CSFR overexpression in pulmonary microvascular endothelial cells and to investigate the effects of different G-CSFR expression levels on downstream SDC4 expression, as well as the activation status of the phosphorylation SRC (pSRC)/nuclear factor-kappaB (NF-κB) pathway.
Results
Compared with the sham group, the LPS group exhibited significantly higher expression levels of tumor necrosis factor-alpha, interleukin-6 (IL-6), and IL-1β in lung tissue, as well as higher inflammatory damage scores; meanwhile, these changes were significantly attenuated by shG-CSFR-mediated gene interference (all P < 0.05). RNA-seq showed that G-CSFR overexpression further increased SDC4 expression [(66 ± 12) in the LPS group vs. (152 ± 19) in the LPS + shG-CSFR group, t = 43.575, q < 0.001], and the Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway involving SDC4 was enriched in the extracellular matrix receptor interaction. LPS stimulation significantly increased SDC4 expression in pulmonary microvascular endothelial cells, and G-CSFR overexpression further enhanced SDC4 messenger RNA and protein in the LPS + shG-CSFR group as compared with the LPS group, accompanied by increased levels of pSRC and phosphorylation pNF-κB/NF-κB (all P < 0.05). Moreover, treatment with PP2 significantly suppressed the expression of pNF-κB/NF-κB and SDC4.
Conclusion
In the context of sepsis-associated ALI, activation and overexpression of G-CSFR in endothelial cells lead to activation of the intracellular pSRC/NF-κB pathway, resulting in overexpression of the inflammatory coordinating factor SDC4, which aggravates inflammation.
To investigate the relationship between peripheral blood microRNA-125a-5p (miR-125a-5p) and miR-483-3p expression and cardiac function, ischemia-reperfusion injury (IRI), and prognosis in patients with acute myocardial infarction (AMI).
Methods
A prospective cohort study was conducted; 136 patients with AMI (AMI group) and 136 healthy volunteers (control group) admitted to Yangling Demonstration Zone Hospital from September 2020 to September 2023 were selected. Patients with AMI were divided into a group Ⅰ-Ⅱ (n = 85) and a group Ⅲ-Ⅳ (n = 51) by Killip class, and also divided into an IRI group (n = 63) and a non-IRI group (n = 73) by IRI occurrence. Patients with AMI were followed up for one year after discharge, with two patients lost. The remaining patients were divided into a good prognosis group (n = 89) and a poor prognosis group (n = 45) according to the prognosis. Peripheral blood miR-125a-5p and miR-483-3p expression levels in all subjects were detected within 24 hours after enrollment, and the differences between groups were compared. Multivariate logistic regression analysis was applied to discover the factors influencing the prognosis of patients with AMI. Based on the analysis results, a combined prediction model was constructed, and a receiver operating characteristic (ROC) curve was used to analyze the predictive value of each indicator individually and jointly in predicting the prognosis of patients with AMI, followed by internal validation.
Results
Peripheral blood miR-125a-5p expression was significantly higher and miR-483-3p expression was significantly lower in the AMI group than in the control group (both P < 0.05), in the group Ⅲ-Ⅳ than in the group Ⅰ-Ⅱ (both P < 0.05), and in the IRI group than in the non-IRI group (both P < 0.05). Killip class Ⅲ-Ⅳ, IRI, and high miR-125a-5p expression were independent risk factors for poor prognosis in patients with AMI, while high miR-483-3p expression was a protective factor (all P < 0.05). The area under the curve (AUC) of peripheral blood miR-125a-5p, miR-483-3p, Killip class Ⅲ-Ⅳ, and IRI for predicting poor prognosis in patients with AMI was 0.785, 0.804, 0.670, and 0.664, and the AUC of the combined prediction model was 0.897. The Delong test demonstrated that the AUC of the combined prediction model was higher than that of peripheral blood miR-125a-5p, miR-483-3p, Killip class Ⅲ-Ⅳ, and IRI alone (Z = 2.637, 2.497, 2.917, 3.017; P = 0.010, 0.014, 0.004, 0.003). The concordance index was 0.684, indicating that the model's predicted results were highly consistent with the actual situation.
Conclusion
High miR-125a-5p and low miR-483-3p expression in peripheral blood are associated with impaired cardiac function, IRI, and poor prognosis in patients with AMI; combining miR-125a-5p, miR-483-3p, Killip class, and IRI can effectively predicting the risk of poor prognosis in these patients.
To investigate the relationship between serum sodium variability within different time windows (0-3 d, 0-7 d, and 0-28 d) after admission to the ICU and the risk of death in patients with cardiac arrest (CA).
Methods
Based on the Medical Information Mart for Intensive Care-Ⅳ of the United States, 6 196 CA patients with complete outcome data were included. Serum sodium variability was calculated respectively in the three time windows of 0-3 days, 0-7 days, and 0-28 days, and grouped according to quartiles (Q1-Q4). The highest quartile (Q4) was regarded as the high variability group, and Q1 to Q3 were combined as the low variability group. Demographic characteristics, admission type, laboratory indicators on the first day of ICU admission, and coagulation sub-item scores and total laboratory scores of sequential organ failure assessment (SOFA) were set as covariates, and serum sodium variability was used as the main exposure variable. The propensity score inverse probability of treatment weighting (IPTW) was used to balance the baseline confounding factors among groups, and logistic regression was performed to analyze the influence of serum sodium variability on the mortality of patients with CA.
Results
Among the 6 196 CA patients, the number of effective analysis cases that met the requirement of having at least two daily mean values of serum sodium after washing for calculable variability was 5 564, 5 590, and 5 624 within the time windows of 0-3 days, 0-7 days, and 0-28 days, respectively. After IPTW correction, in the 0-28 days window, the risk of death in the high variability group was significantly higher than that in the low variability group [odds ratio (OR) = 1.950, 95% confidence interval (CI) (1.700, 2.238), P < 0.001]; the results of 0-3 days [OR = 1.411, 95%CI (1.117, 1.782), P = 0.004] and 0-7 days [OR = 1.579, 95%CI (1.335, 1.868), P < 0.001] time windows showed consistent trends.
Conclusion
The high serum sodium variability within different time windows after ICU admission is associated with an increased mortality risk in CA patients, suggesting that the fluctuation range of serum sodium can be used as a reference indicator for prognostic risk stratification in these patients.
To evaluate the non-linear association and threshold effect between serum anion gap (AG) levels on the first day of ICU admission and the risk of delirium during the ICU stay in acute pancreatitis (AP) patients.
Methods
This retrospective study utilized the medical information mart for intensive care-Ⅳ (MIMIC-Ⅳ) database, including adult AP patients admitted to the ICU for the first time between 2008 and 2019. The primary exposure variable was the AG value first measured within 24 hours of ICU admission and the primary outcome was the occurrence of delirium during the ICU stay. Multivariate logistic regression models, adjusting for demographic characteristics, disease severity, and other key clinical variables, were used to assess the independent association between AG and delirium. Generalized additive models (GAM) and two-piecewise linear regression models were employed to explore its non-linear relationship and potential threshold.
Results
A total of 890 AP patients were included, of whom 237 (26.6%) experienced delirium during their ICU stay. After multivariate logistic regression models adjustment, the overall AG was not significantly associated with the delirium risk [odds ratio (OR) = 1.028, 95% confidence interval (CI) (0.993, 1.064), P = 0.117]. However, GAM and two-piecewise linear regression analysis revealed a significant non-linear relationship between AG and delirium, with a threshold of 14.0 mmol/L. Specifically, when AG was < 14.0 mmol/L, an increase of 1.0 mmol/L in AG was associated with a 15.5% increased risk of delirium [OR = 1.155, 95%CI (1.018, 1.310), P = 0.025]; when AG was ≥ 14.0 mmol/L, the association was not statistically significant [OR = 1.005, 95%CI (0.963, 1.048), P = 0.827].
Conclusions
Serum AG levels on the first day of ICU admission in AP patients are non-linearly associated with the risk of delirium during ICU stay, with a positive correlation only present when AG is below approximately 14.0 mmol/L. AG, as a routine, convenient, and cost-effective metabolic indicator, has potential value in early delirium risk stratification for AP patients, offering a reference for clinical warning and targeted interventions.
Chinese Society of Physical Medicine and Rehabilitation, Visceral Rehabilitation Group, Rehabilitation Physicians Branch of Chinese Medical Doctor Association, Health Care Working Committee of Chinese Association of Rehabilitation Medicine