[Effect of PFKFB3 on inflammatory activation of polymorphonuclear myeloid-derived suppressor cell in acute myocardial infarction].

OBJECTIVE: To investigate the correlation between 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3 (PFKFB3) and the inflammatory activation of polymorphonuclear myeloid-derived suppressor cell (PMN-MDSC) in acute myocardial infarction (AMI), and to evaluate the effect of intervention targeting PFKFB3 on the inflammatory activation of PMN-MDSC during AMI. METHODS: (1) Clinical trial section: a observational study was conducted. The patients with acute coronary syndrome (ACS) admitted to Zhenjiang Fourth People's Hospital were enrolled, and they were divided into AMI group and non-AMI group according to clinical diagnosis. The peripheral venous blood of the two groups was collected to detect the proportion of PMN-MDSC, and the expression of PFKFB3 gene in mononuclear cells was detected by real-time quantitative polymerase chain reaction (RT-qPCR). (2) Basic experiment section: a total of 30 male C57 mice (aged 6-8 weeks) were divided into normal control group (n = 5), Sham group (n = 5), AMI model group (n = 10) and PFKFB3 inhibitor PKF-15 intervention group (n = 10) according to random number table method. The AMI model of mice was reproduced by left anterior descending coronary artery (LADCA) ligation, and the mice in the Sham group did not attach the artery after thoracotomy. The PKF-15 intervention group was intraperitoneally injected with PKF-15 (20 µg/g) at the same time of LADCA ligation. Normal control mice did not receive any treatment. Peripheral venous blood and myocardial tissue of mice were collected 24 hours after modeling. Both the circulating PMN-MDSC ratio and the infiltration of PMN-MDSC in myocardial tissue were detected. After staining with hematoxylin-eosin (HE), the degree of inflammatory damage in mouse myocardial tissue was observed under light microscopy. PMN-MDSC were isolated from mice with flow cytometry, and the gene expressions of PFKFB3 and inflammatory factors were measured by RT-qPCR. RESULTS: (1) Clinical trial section: the circulating PMN-MDSC ratio of patients in the AMI group (n = 25) was significantly higher than that in the non-AMI group [n = 20; (8.53±0.96)% vs. (1.13±0.39)%, P < 0.01], and PFKFB3 gene expression in the peripheral blood mononuclear cells was also increased (2-ΔΔCt: 1.18±0.19 vs. 0.96±0.16, P < 0.01). Pearson correlation analysis showed that circulating PMN-MDSC ratio was positively correlated with PFKFB3 gene expression in mononuclear cells in AMI patients (r = 0.608, P = 0.001). (2) Basic experimental section: the circulating PMN-MDSC ratio and the infiltration of PMN-MDSC in myocardial tissue of AMI mice were significantly higher than those in the normal control group and Sham group. PFK-15 intervention could reduce the ratio of PMN-MDSC in the peripheral blood and myocardial tissue of AMI mice [(26.33±5.27)% vs. (75.12±5.02)% in peripheral blood, (20.87±2.97)% vs. (35.28±4.36)% in myocardial tissue, both P < 0.01]. Under light microscopy, the myocardial cells in the AMI modal group were disordered and a large number of inflammatory cells infiltrated. PFK-15 intervention could maintain a normal arrangement of cardiomyocytes and reduce the infiltration of inflammatory cells. The gene expression levels of PFKFB3 in the peripheral blood and myocardial tissue as well as the inflammatory factors in the myocardial tissue of AMI mice were significantly higher than those in the normal control group and Sham group. PKF-15 intervention could effectively reduce the gene expression levels of PFKFB3 in the peripheral blood and myocardial tissue as well as the inflammatory factors in the myocardial tissue of AMI mice [PFKFB3 mRNA (2-ΔΔCt): 1.01±0.09 vs. 1.40±0.12 in peripheral blood, 0.95±0.09 vs. 1.47±0.10 in myocardial tissue; myocardial tissue tumor necrosis factor-α (TNF-α) mRNA (2-ΔΔCt) was 14.55±3.99 vs. 29.66±3.90, interleukin-1ß (IL-1ß) mRNA (2-ΔΔCt) was 8.72±1.35 vs. 18.53±2.43, IL-6 mRNA (2-ΔΔCt) was 11.87±2.97 vs. 19.82±4.32, all P < 0.01]. CONCLUSIONS: The activation of PFKFB3 is closely related to the inflammatory activation of PMN-MDSC during AMI. Inhibition of PFKFB3 activity can inhibit the inflammatory activation of PMN-MDSC and reduce myocardial inflammatory injury.

Blocking the A2B adenosine receptor alleviates myocardial damage by inhibiting spleen-derived MDSC mobilisation after acute myocardial infarction.

BACKGROUND: Myeloid-derived suppressor cell (MDSC) mobilisation is an important immune event in acute myocardial infarction (AMI). The A2B adenosine receptor (A2BAR) plays key role in regulating MDSC function, but its specific involvement in MDSC mobilisation in AMI remains unclear. METHODS: In AMI patients, the circulating MDSC ratio and A2BAR mRNA expression were measured. A mouse AMI model was established by left anterior descending coronary artery (LADCA) ligation. MDSCs were analysed by FACS and immunofluorescence staining (of heart tissue). A2BAR mRNA expression was assessed by qRT-PCR. Myocardial injury was detected by HE staining. Myocardial cell apoptosis was analysed by immunohistochemistry. Cardiac systolic function was evaluated by transthoracic echocardiography. RESULTS: In AMI patients, the circulating MDSC ratio was increased and positively correlated with A2BAR mRNA expression (r = 0.86, p < 0.01). In AMI model mice, the percentage of MDSCs was increased in the circulation and infarcted heart and decreased in the spleen. MRS-1754-mediated A2BAR inhibition decreased the MDSC ratio in the circulation and infarcted heart and prevented the decrease in MDSC number in the spleens of mice with AMI. A2BAR blockade inhibited myocardial cell apoptosis, alleviated myocardial inflammatory injury, and improved myocardial systolic function in the AMI mouse model. Similar results were found in mice after splenectomy. Additionally, spleen-derived MDSC injection increased the MDSC ratio in the infarcted heart, increased myocardial cell apoptosis, aggravated myocardial injury, and decreased cardiac systolic function in mice with AMI. CONCLUSION: Blocking A2BAR alleviates myocardial damage and improves myocardial systolic function through inhibition of spleen-derived MDSC mobilisation after AMI. Key MessagesSpleen-derived MDSC mobilisation aggravates myocardial inflammatory injury within 24 h of AMI.A2BAR promotes spleen-derived MDSC mobilisation within 24 h of AMI.Blocking A2BAR improves myocardial systolic function through inhibition of spleen-derived MDSC mobilisation.

Long noncoding RNA GSEC promotes neutrophil inflammatory activation by supporting PFKFB3-involved glycolytic metabolism in sepsis.

Metabolic reprogramming is a hallmark of neutrophil activation in sepsis. LncRNAs play important roles in manipulating cell metabolism; however, their specific involvement in neutrophil activation in sepsis remains unclear. Here we found that 11 lncRNAs and 105 mRNAs were differentially expressed in three transcriptome datasets (GSE13904, GSE28750, and GSE64457) of gene expression in blood leukocytes and neutrophils of septic patients and healthy volunteers. After Gene Ontology biological process analysis and lncRNA-mRNA pathway network construction, we noticed that GSEC lncRNA and PFKFB3 were co-expressed and associated with enhanced glycolytic metabolism. Our clinical observations confirmed the expression patterns of GSEC lncRNA and PFKFB3 genes in neutrophils in septic patients. Performing in vitro experiments, we found that the expression of GSEC lncRNA and PFKFB3 was increased when neutrophils were treated with inflammatory stimuli. Knockdown and overexpression experiments showed that GSEC lncRNA was essential for mediating PFKFB3 mRNA expression and stability in neutrophil-like dHL-60 cells. In addition, we found that GSEC lncRNA-induced PFKFB3 expression was essential for mediating dHL-60 cell inflammatory cytokine expression. Performing mechanistic experiments, we found that glycolytic metabolism with PFKFB3 involvement supported inflammatory cytokine expression. In summary, our study uncovers a mechanism by which GSEC lncRNA promotes neutrophil inflammatory activation in sepsis by supporting glycolytic metabolism with PFKFB3.

[Value of neutrophil to lymphocytes and platelets ratio for predicting 28-day mortality in sepsis patients].

OBJECTIVE: To evaluate the value of neutrophil to lymphocyte and platelet ratio (N/LPR) for predicting 28-day mortality in sepsis patients. METHODS: A retrospective analysis was conducted. The clinical data of 154 sepsis patients admitted to intensive care unit (ICU) of the Affiliated Hospital of Jiangsu University from June 2017 to June 2020 were enrolled. The time of first diagnosis of sepsis in ICU was taken as the research starting point, and the death or 28 days as the end point. The 28-day outcomes of patients were recorded. The counts of peripheral blood neutrophil (NEU), lymphocyte (LYM) and platelet (PLT) were collected from all the enrolled patients within 3 days after diagnosis of sepsis. The ratios of N/LPR and NEU/LYM (NLR) were calculated respectively. The differences of N/LPR and NLR between survival group and death group were compared. Receiver operating characteristic (ROC) curve analysis was used to analyze the value of N/LPR and NLR on predicting the 28-day mortality of sepsis patients. According to the best cut-off value of ROC curve analysis, the 28-day mortality of patients with sepsis was analyzed by subgroup analysis, and the 28-day cumulative survival of patients with sepsis was analyzed by Kaplan-Meier survival curve. RESULTS: Of the 154 sepsis patients, the patients with age < 18 years, pregnancy, blood disease, taking aspirin or other antiplatelet drugs within 1 week, taking leucocyte drugs within 1 week, length of ICU stay < 3 days and incomplete data were excluded. Finally, 50 patients were enrolled. Among them, 30 patients survived on the 28th day and 20 died. Compared with the survival group, the levels of N/LPR and NLR in the death group were significantly increased (N/LPR: 23.85±11.99 vs. 12.41±5.25, NLR: 17.83±8.69 vs. 10.75±3.63), with statistical differences (both P < 0.01). ROC curve analysis indicated that the area under ROC curve (AUC) of N/LPR for predicting 28-day death of sepsis patients was 0.827, it was higher than that of NLR (AUC = 0.762). Base on N/LPR ≥ 15.48 as a predictor of cut-off value of death in 28 days of sepsis patients, the sensitivity was 75.0% and the specificity was 80.0%, respectively. Base on NLR ≥ 10.65 as a predictor of cut-off value of death in 28 days of sepsis patients, the sensitivity was 75.0% and specificity was 56.7%, respectively. Subgroup analysis showed that the 28-day mortality in the patients with N/LPR ≥ 15.48 (n = 21) was significantly higher than those with N/LPR < 15.48 (n = 29; 71.4% vs. 17.2%, χ2 = 14.901, P < 0.01); and the 28-day mortality in the patients with NLR ≥ 10.65 (n = 28) was also significantly higher than those with NLR < 10.65 (n = 22; 53.6% vs. 22.7%, χ2 = 4.884, P < 0.05). The results were consistent with Kaplan-Meier survival curve analysis. CONCLUSIONS: Peripheral blood N/LPR has a good predictive value for 28-day mortality of sepsis patients, and which is better than NLR.

Peguero-Lo Presti criteria for the diagnosis of left ventricular hypertrophy: A systematic review and meta-analysis.

BACKGROUND: The Peguero-Lo Presti criteria are novel electrocardiographic (ECG) diagnostic criteria for the detection of left ventricular hypertrophy (LVH) and represent the sum of the amplitude of the deepest S wave in any lead with the S wave in lead V4 (SD+SV4). The diagnostic efficacy of the Peguero-Lo Presti criteria in LVH is still debatable. We aimed to test the sensitivity and specificity of the Peguero-Lo Presti criteria and compared them with those of the Cornell voltage index to assess their overall performance in LVH diagnosis. METHODS: Electronic databases (e.g., Medline, Web of Knowledge, Embase, and the Cochrane Library) were searched from their inception until May 18, 2020. Trials written in English that investigated the Peguero-Lo Presti criteria for detecting LVH were included. Data were independently extracted and analyzed by two investigators. RESULTS: A total of 51 records were screened, and 6 trials comprising 13,564 patients were finally included. A bivariate analysis showed that the sensitivity of the Peguero-Lo Presti criteria (0.52, 95% confidence interval (CI) 0.46-0.58) was higher than that of the Cornell voltage index (0.29, 95% CI 0.23-0.36) and Sokolow-Lyon criteria (0.24, 95% CI 0.21-0.27); the diagnostic accuracy of the Peguero-Lo Presti criteria (0.69, 95% CI 0.65-0.73) was also higher than that of the Cornell voltage index (0.67, 95% CI 0.62-0.71) and Sokolow-Lyon criteria (0.28, 95% CI 0.25-0.32); and the specificity of the Peguero-Lo Presti criteria (0.85, 95% CI 0.79-0.90) was similar to that of the Cornell voltage index (0.92, 95% CI 0.89-0.95) and Sokolow-Lyon criteria (0.94, 95%CI 0.88-0.97). Two trials (including 12,748 patients) were discharged because they included partly healthy subjects and accounted for substantial heterogeneity. Pooled analysis of the remaining 4 trials (including 816 patients) showed that the sensitivity of the Peguero-Lo Presti criteria (0.56, 95% CI 0.51-0.61) was also higher than that of the Cornell voltage index (0.36, 95% CI 0.31-0.42) and Sokolow-Lyon criteria (0.24, 95% CI 0.18-0.31); the diagnostic accuracy of the Peguero-Lo Presti criteria (0.84, 95% CI 0.80-0.87) was also higher than that of the Cornell voltage index (0.54, 95% CI 0.50-0.58) and Sokolow-Lyon criteria (0.38, 95% CI 0.34-0.42); and the specificity of the Peguero-Lo Presti criteria (0.90, 95% CI 0.87-0.92) was similar to that of the Cornell voltage index (0.93, 95% CI 0.88-0.96) and Sokolow-Lyon criteria (0.97, 95% CI 0.90-0.99). Both the likelihood ratio and posttest probability of the Peguero-Lo Presti criteria and Cornell voltage index were moderate. CONCLUSION: Based on this systematic review and meta-analysis, the Peguero-Lo Presti criteria-based ECG diagnostic method for LVH has high sensitivity, specificity and diagnostic accuracy and should be applied in clinical practice settings.

Effect of ulinastatin combined with thymosin alpha1 on sepsis: A systematic review and meta-analysis of Chinese and Indian patients.

PURPOSE: To assess the effects of urinary trypsin inhibitor (UTI) ulinastatin combined with thymosin alpha1 (Tα1) on sepsis. MATERIALS AND METHODS: The meta-analysis included 8 randomized controlled trials (N=1112 patients) on UTI-based therapy for sepsis published before July 10, 2016. Two investigators independently extracted data and assessed the quality of each study. The short-term mortality rate, duration of mechanical ventilator and vasopressor use, length of intensive care unit stay, Acute Physiology and Chronic Health Evaluation (APACHE) II score, and differences in inflammatory cytokines (interleukin [IL]-6, IL-10, and tumor necrosis factor α) were assessed using statistical software. RESULTS: Treatment of UTI combined with Tα1 (UTI+Tα1) decreased the short-term mortality rate in septic patients by 36%, 35%, and 31% for 28, 60, 90 days, respectively. UTI+Tα1 decreased the duration of mechanical ventilation and vasopressor use, APACHE II score, and levels of IL-6 and tumor necrosis factor α. Treatment of UTI+Tα1 did not reduce the duration of vasopressor use and length of intensive care unit stay, or increase IL-10 levels. Because of the high heterogeneity of the included trials, the results should be carefully assessed. CONCLUSIONS: Treatment of UTI+Tα1 can suppress the production of proinflammatory cytokines, decrease the APACHE II score, shorten the duration of mechanical ventilation and vasopressor use, and improve the 28-day survival rate.

Effect of ulinastatin combined with thymosin alpha1 on sepsis: A systematic review and meta-analysis of Chinese and Indian patients.

PURPOSE: To assess the effects of urinary trypsin inhibitor (UTI) ulinastatin combined with thymosin alpha1 (Tα1) on sepsis. MATERIALS AND METHODS: The meta-analysis included 8 randomized controlled trials (N=1112 patients) on UTI-based therapy for sepsis published before July 10, 2016. Two investigators independently extracted data and assessed the quality of each study. The short-term mortality rate, duration of mechanical ventilator and vasopressor use, length of intensive care unit stay, Acute Physiology and Chronic Health Evaluation (APACHE) II score, and differences in inflammatory cytokines (interleukin [IL]-6, IL-10, and tumor necrosis factor α) were assessed using statistical software. RESULTS: Treatment of UTI combined with Tα1 (UTI+Tα1) decreased the short-term mortality rate in septic patients by 36%, 35%, and 31% for 28, 60, 90 days, respectively. UTI+Tα1 decreased the duration of mechanical ventilation, APACHE II score, and levels of IL-6 and tumor necrosis factor α. Treatment of UTI+Tα1 did not reduce the duration of vasopressor use and length of intensive care unit stay, or increase IL-10 levels. Because of the high heterogeneity of the included trials, the results should be carefully assessed. CONCLUSIONS: Treatment of UTI+Tα1 can suppress the production of proinflammatory cytokines, decrease the APACHE II score, shorten the duration of mechanical ventilation, and improve the 28-day survival rate.

A large-scale measurement of electromagnetic fields near GSM base stations in Guangxi, China for risk communication.

Radiofrequency (RF) electromagnetic field (EMF) exposure from wireless telecommunication base station antennae can lead to debates, conflicts or litigations among the adjacent residents if inappropriately managed. This paper presents a measurement campaign for the GSM band EMF exposure in the vicinity of 827 base station sites (totally 6207 measurement points) in Guangxi, China. Measurement specifications are designed for risk communication with the residents who previously complained of over-exposure. The EMF power densities with the global positioning system coordinate at each measured point were recorded. Compliance with the International Commission on Non-Ionizing Radiation Protection guidelines and Chinese environmental EMF safety standards was studied. The results show that the GSM band EMF level near the base stations is very low. The measurement results and the EMF risk communication procedures positively influence public perception of the RF EMF exposure from the base stations and promote the exchange of EMF exposure-related knowledge.