Alpha-Lipoic Acid Suppresses Extracellular Histone-Induced Release of the Infammatory Mediator Tumor Necrosis Factor-α by Macrophages.

BACKGROUND/AIMS: This study investigated signaling pathways via which extracellular histones induce the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) release from the macrophage cell line RAW 264.7 and the anti-inflammatory efficacy of the antioxidant alpha-lipoic acid (ALA). METHODS: ELISA and western blotting analyses were conducted to detect the release of TNF-α from histone-stimulated RAW 264.7 macrophages and the associated phospho-activation of MAPKs (ERK and p38) and NF-κB p65. The effects of ALA on the release of TNF-α and phospho-activation of the MAPKs and NF-κB p65 were studied. P < 0.05 was considered statistically significant. RESULTS: Extracellular histones dose-dependently induced TNF-α release from RAW 264.7 cells and increased the phosphorylation of p38, ERK, and NF-κB p65. TNF-α release was markedly suppressed by p38, ERK, and NF-kB inhibitors. ALA reduced histone-induced TNF-α release, ERK/p38 MAPK activation, and NF-kB activation without affecting macrophage viability. CONCLUSION: Histones induce TNF-α release from macrophages by activating the MAPK and NF-kB signaling pathways, while ALA suppresses this response by inhibiting ERK, p38 and NF-kB. These findings identify potentially critical inflammatory signaling pathways in sepsis and molecular targets for sepsis treatment.

NEMO-Binding Domain Peptide Attenuates Lipopolysaccharide-Induced Acute Lung Injury by Inhibiting the NF-κB Signaling Pathway.

The aim of the present study is to investigate the protective effects and relevant mechanisms exerted by NEMO-binding domain peptide (NBD) against lipopolysaccharide- (LPS-) induced acute lung injury (ALI) in mice. The ALI model was induced by intratracheally administered atomized LPS (5 mg/kg) to BABL/c mice. Half an hour before LPS administration, we treated the mice with increasing concentrations of intratracheally administered NBD or saline aerosol. Two hours after LPS administration, each group of mice was sacrificed. We observed that NBD pretreatment significantly attenuated LPS-induced lung histopathological injury in a dose-dependent manner. Western blotting established that NBD pretreatment obviously attenuated LPS-induced IκB-α and NF-κBp65 activation and NOX1, NOX2, and NOX4 overexpression. Furthermore, NBD pretreatment increased SOD and T-AOC activity and decreased MDA levels in lung tissue. In addition, NBD also inhibited TNF-α and IL-1ß secretion in BALF after LPS challenge. In conclusion, NBD protects against LPS-induced ALI in mice.

MiR-137 inhibits proliferation and angiogenesis of human glioblastoma cells by targeting EZH2.

It is suggested that microRNAs play important roles in the development of various cancers. Here, we showed that miR-137 is downregulated in glioblastoma (GBM) cell lines and that low levels of miR-137 are associated with a poor prognostic phenotype of GBM patients. Ectopic expression of miR-137 significantly inhibited GBM cell proliferation and angiogenesis. In addition, ectopic expression of miR-137 inhibited tumor growth and angiogenesis in a SCID mouse xenograft model. EZH2 was identified as a direct target of miR-137 by using luciferase reporter and Western blot assays, and EZH2 overexpression can rescue the inhibitory effect of miR-137 on cell proliferation and angiogenesis. Furthermore, tumor samples from GBM patients showed an inverse relationship between miR-137 and EZH2 levels. Our results suggest that miR-137 may serve as a biomarker in GBM, and the modulation of its activity may represent a novel therapeutic strategy for the treatment of GBM patients.

miR-122-3p targets UBE2I to regulate the immunosuppression of liver cancer and the intervention of Liujunzi formula.

ETHNOPHARMACOLOGICAL RELEVANCE: Liujunzi formula has been used to treat liver cancer in China for many years, but its underlying mechanism remains unclear. We previously found that decreased expression of miR-122-3p was associated with liver cancer. In this study, we aimed to explore the target of miR-122-3p and the effect of the Liujunzi formula on miR-122-3p and its downstream events in liver cancer. MATERIAL AND METHODS: Bioinformatics pinpointed potential targets of miR-122-3p. The actual target was confirmed by miRNA mimic/inhibitor transfections and a dual-luciferase reporter assay. RNA-seq looked at downstream genes impacted by this target. Flow cytometry checked for changes in T cell apoptosis levels after exposing them to liver cancer cells. Gene expression was measured by RT-qPCR, western blotting, and immunofluorescence staining. RESULTS: Cell experiments found the Liujunzi extract (LJZ) upregulated miR-122-3p and in a dose-dependent manner. Bioinformatics analysis found UBE2I was a potential target of miR-122-3p, which was validated through experiments using miRNA mimics/inhibitors and a dual-luciferase reporter assay. RNA-seq data implicated the NF-κB pathway as being downstream of the miR-122-3p/UBE2I axis, further confirmed by forcing overexpression of UBE2I. Bioinformatic evidence suggested a link between UBE2I and T cell infiltration in liver cancer. Given that the NF-κB pathway drives PD-L1 expression, which can inhibit T cell infiltration, we investigated whether PD-L1 is a downstream effector of miR-122-3p/UBE2I. This was corroborated through mining public databases, UBE2I overexpression studies, and tumor-T cell co-culture assays. In addition, we also confirmed that LJZ downregulates UBE2I and NF-κB/PD-L1 pathways through miR-122-3p. LJZ also suppressed SUMOylation in liver cancer cells and protected PD-1+ T cells from apoptosis induced by co-culture with tumor cells. Strikingly, a miR-122-3p inhibitor abrogated LJZ's effects on UBE2I and PD-L1, and UBE2I overexpression rescued the LJZ-mediated effects on NF-κB and PD-L1. CONCLUSIONS: miR-122-3p targets UBE2I, thereby suppressing the NF-κB signaling cascade and downregulating PD-L1 expression, which potentiates anti-tumor immune responses. LJZ bolsters anti-tumor immunity by modulating the miR-122-3p/UBE2I/NF-κB/PD-L1 axis in liver cancer cells.

Single-Channel sEMG-Based Estimation of Knee Joint Angle Using a Decomposition Algorithm With a State-Space Model.

Accurate human motion estimation is crucial for effective and safe human-robot interaction when using robotic devices for rehabilitation or performance enhancement. Although surface electromyography (sEMG) signals have been widely used to estimate human movements, conventional sEMG-based methods, which need sEMG signals measured from multiple relevant muscles, are usually subject to some limitations, including interference between sEMG sensors and wearable robots/environment, complicated calibration, as well as discomfort during long-term routine use. Few methods have been proposed to deal with these limitations by using single-channel sEMG (i.e., reducing the sEMG sensors as much as possible). The main challenge for developing single-channel sEMG-based estimation methods is that high estimation accuracy is difficult to be guaranteed. To address this problem, we proposed an sEMG-driven state-space model combined with an sEMG decomposition algorithm to improve the accuracy of knee joint movement estimation based on single-channel sEMG signals measured from gastrocnemius. The effectiveness of the method was evaluated via both single- and multi-speed walking experiments with seven and four healthy subjects, respectively. The results showed that the normal root-mean-squared error of the estimated knee joint angle using the method could be limited to 15%. Moreover, this method is robust with respect to variations in walking speeds. The estimation performance of this method was basically comparable to that of state-of-the-art studies using multi-channel sEMG.

Quality of life and mortality in older adults with sepsis after one-year follow up: A prospective cohort study demonstrating the significant impact of frailty.

BACKGROUND: Survivors of sepsis often develop chronic critical illness after the inflammatory stage, resulting in death or hospital readmission. The long-term prognosis of older patients with sepsis and the associated factors, particularly frailty, are not well studied. OBJECTIVES: To investigate the effect of frailty on the quality of life (QoL) and mortality in older patients after one year of sepsis diagnosis. METHODS: This prospective study included patients admitted to a specialized geriatric intensive care unit between May 2018 and April 2019. Patients were grouped according to the Clinical Frailty Scale as severely frail, mildly to moderately frail, and non-frail/vulnerable. The primary outcome was QoL one year after sepsis diagnosis, measured using the European QoL 5-Dimension (EQ-5D) and 12-item Short Form. The secondary outcome was one-year survival. RESULTS: Of the 211 participants, 75 (35.5%) completed the QoL surveys. Of them, 37 (49.3%) did not return to their baseline QoL one year after sepsis diagnosis. The rate of reported mobility problems (a dimension of the EQ-5D) increased by 100% during the year. Additionally, survivors in the severely frail group exhibited poorer QoL at one year than those in the mildly to moderately frail and non-frail/vulnerable groups. The one-year mortality in the severely frail group was 75.9%, with an adjusted hazard ratio of 1.70 (95% confidence interval, 1.02-2.82, p = 0.041). CONCLUSIONS: Frailty significantly impacts the one-year prognosis in older patients with sepsis. This research highlights the need for frailty management and physical rehabilitation in frail older patients at risk of poor prognosis, with implications for improving transitional and post-acute care services.

C-reactive protein-to-albumin ratio as a biomarker in patients with sepsis: a novel LASSO-COX based prognostic nomogram.

To develop a C-reactive protein-to-albumin ratio (CAR)-based nomogram for predicting the risk of in-hospital death in sepsis patients. Sepsis patients were selected from the MIMIC-IV database. Independent predictors were determined by multiple Cox analysis and then integrated to predict survival. The performance of the model was evaluated using the concordance index (C-index), receiver operating characteristic curve (ROC) analysis, and calibration curve. The risk stratifications analysis and subgroup analysis of the model in overall survival (OS) were assessed by Kaplan-Meier (K-M) curves. A total of 6414 sepsis patients were included. C-index of the CAR-based model was 0.917 [standard error (SE): 0.112] for the training set and 0.935 (SE: 0.010) for the validation set. The ROC curve analysis showed that the area under the curve (AUC) of the nomogram was 0.881 in the training set and 0.801 in the validation set. And the calibration curve showed that the nomogram performs well in both the training and validation sets. K-M curves indicated that patients with high CAR had significantly higher in-hospital mortality than those with low CAR. The CAR-based model has considerably high accuracy for predicting the OS of sepsis patients.

Analysis of Pharmacological Activities and Mechanisms of Essential Oil in Leaves of C. grandis 'Tomentosa' by GC-MS/MS and Network Pharmacology.

BACKGROUND: Citrus grandis 'Tomentosa,' a fruit epicarp of C. grandis 'Tomentosa' or C. grandis (L.) Osbeck is widely used in health food and medicine. Based on our survey results, there are also rich essential oils with bioactivities in leaves, but the chemical compounds in this part and relevant pharmacological activities have never been studied systematically. Therefore, this study was to preliminarily decipher the pharmacological activities and mechanisms of the essential oil in leaves of C. grandis 'Tomentosa' by an integrated network pharmacology approach. METHODS: Essential oil compositions from leaves ofC. grandis 'Tomentosa' were identified using GC-MS/MS. And then, the targets of these oil compositions were predicted and screened from TCMSP, SwissTargetPrediction, STITCH and SEA databases. STRING database was used to construct the protein-protein interaction networks, and the eligible protein targets were input into WebGestalt 2019 to carry out GO enrichment and KEGG pathway enrichment analysis. Based on the potential targets, disease enrichment information was obtained by TTD databases. Cytoscape software was used to construct the component-target-disease network diagrams. RESULTS: Finally, 61 essential oil chemical components were identified by GC-MS/MS, which correspond to 679 potential targets. Biological function analysis showed 12, 19, and 12 GO entries related to biological processes, cell components and molecular functions, respectively. 43 KEGG pathways were identified, of which the most significant categories were terpenoid backbone biosynthesis, TNF signaling pathway and leishmaniasis. The component-target-disease network diagram revealed that the essential oil compositions in leaves of C. grandis 'Tomentosa' could treat tumors, immune diseases, neurodegenerative diseases and respiratory diseases, which were highly related to CHRM1, PTGS2, CASP3, MAP2K1 and CDC25B. CONCLUSION: This study may provide new insight into C. grandis 'Tomentosa' or C. grandis (L.) Osbeck and may provide useful information for future utilization and development.

A reciprocal inhibitory module for Pi and iron signaling.

Phosphorous (P) and iron (Fe), two essential nutrients for plant growth and development, are highly abundant elements in the earth's crust but often display low availability to plants. Due to the ability to form insoluble complexes, the antagonistic interaction between P and Fe nutrition in plants has been noticed for decades. However, the underlying molecular mechanism modulating the signaling and homeostasis between them remains obscure. Here, we show that the possible iron sensors HRZs, the iron deficiency-induced E3 ligases, could interact with the central regulator of phosphate (Pi) signaling, PHR2, and prompt its ubiquitination at lysine residues K319 and K328, leading to its degradation in rice. Consistent with this, the hrzs mutants displayed a high Pi accumulation phenotype. Furthermore, we found that iron deficiency could attenuate Pi starvation signaling by inducing the expression of HRZs, which in turn trigger PHR2 protein degradation. Interestingly, on the other hand, rice PHRs could negatively regulate the expression of HRZs to modulate iron deficiency responses. Therefore, PHR2 and HRZs form a reciprocal inhibitory module to coordinate Pi and iron signaling and homeostasis in rice. Taken together, our results uncover a molecular link between Pi and iron master regulators, which fine-tunes plant adaptation to Pi and iron availability in rice.

Study on the mechanism of Cortex Lycii on lung cancer based on network pharmacology combined with experimental validation.

ETHNOPHARMACOLOGICAL RELEVANCE: Xie Bai San is a Chinese medicine prescription that has been used to treat lung cancer in China for a long time. It has been proven to alleviate the symptoms and extend the survival time of lung cancer patients. Xie Bai San comprises Cortex Lycii, Cortex Mori, and Radix Glycyrrhizae Preparata. The effects and mechanisms of Cortex Mori and Glycyrrhizae on lung cancer have been reported, whereas the underlying mechanism of Cortex Lycii remains unknown. MATERIAL AND METHODS: Network pharmacology was used to explore the unknown mechanisms underlying the effect of Cortex Lycii on lung cancer. Molecular docking was used to predict the binding of a compound to the protein. The fingerprint of Cortex Lycii was obtained by HPLC. Cell counting Kit-8 assay was used to determine the appropriate concentration of Cortex Lycii extract for human lung adenocarcinoma cells, A549 and H1299. Wound healing assay and Matrigel invasion assay were used to detect the influence of Cortex Lycii extract on the migration and invasion ability of A549 and H1299. The protein expression level was detected by western blot and immunohistochemical staining. RESULTS: Using network pharmacology, 38 components of Cortex Lycii and 79 possible lung cancer-related target genes of Cortex Lycii were obtained. The targets were assigned to 35 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and the PI3K-AKT signaling pathway contained the most targets and had the second-lowest P-value. The molecular docking showed the components of Cortex Lycii bound to HSP90AB1. Among them, 6 components bound to HSP90AB1 in which HSP90AB1 binds to and phosphorylates AKT. The functional experiments showed that Cortex Lycii suppressed the migration and invasion of human lung cancer cells in a dose-dependent manner. Cortex Lycii up-regulated E-Cadherin and down-regulated N-Cadherin, Vimentin, and MMP2. Furthermore, Cortex Lycii made no change in the total AKT and mTOR protein levels, but caused the down-regulation of p-AKT and p-mTOR in human lung cancer cells, which was reversed by Terazosin, an agonist of HSP90. Moreover, acacetin and apigenin, two components of Cortex Lycii, reduced the protein level of p-AKT and p-mTOR, and the reduction was also inhibited by Terazosin. CONCLUSION: Cortex Lycii suppressed epithelial-mesenchymal transition (EMT) in lung cancer cells through the PI3K-AKT-mTOR signaling pathway, possibly by targeting HSP90AB1 and inhibiting HSP90AB1-AKT binding.

[Predictive value of neutrophil to lymphocyte ratio in the progression of sepsis to chronic critical illness in elderly patients].

OBJECTIVE: To explore the predictive value of neutrophil to lymphocyte ratio (NLR) in the progression of sepsis to chronic critical illness (CCI) in elderly patients. METHODS: Patients with sepsis who were hospitalized more than 24 hours and older than 60 years old admitted to the department of medical intensive care unit (MICU) of General Hospital of Southern Theatre Command from August 2019 to April 2021 were enrolled. The neutrophil count (NEU), lymphocyte count (LYM) and NLR of peripheral blood cells were recorded on the 1st, 4th and 7th day after admission. Patients were divided into the CCI group and the non-CCI group according to whether they progressed to CCI, and differences between the two groups were compared. The CCI was defined as a MICU length of stay (LOS) ≥ 14 days and persistent organ dysfunction [sequential organ failure assessment (SOFA) score ≥ 2]. Logistic regression analysis was performed to evaluate the risk factors for predicting CCI. The receiver operator characteristic curve (ROC curve) was plotted for evaluating the predictive value of NLR in the progression of sepsis to CCI in elderly patients. RESULTS: (1) Among 103 sepsis patients enrolled, 16 (15.5%) died within 2 weeks of admission to the MICU, 46 (44.7%) developed CCI, and 41 (39.8%) were non-CCI. (2) Compared between the two groups, the NEU of CCI group on day 7 was significantly higher than that of non-CCI group [×109/L: 9.80 (6.72, 16.80) vs. 6.66 (5.14, 9.29), P < 0.01], LYM was significantly lower than that of non-CCI group [×109/L: 0.77 (0.46, 1.20) vs. 1.00 (0.86, 1.48), P < 0.05], and NLR on day 4 and day 7 were significantly higher than those of non-CCI group [12.85 (6.56, 17.56) vs. 8.26 (5.34, 13.17), 13.76 (6.97, 23.66) vs. 6.14 (4.04, 8.84), both P < 0.05]. Compared with different time points in the same group, NEU and NLR decreased gradually and LYM increased gradually in non-CCI group (χ2 values were 10.216, 28.343, 7.189, respectively, all P < 0.05), which tended to be normal. There were no significant differences in NEU, LYM and NLR of CCI group at each time point (χ2 values were 0.798, 4.478, 5.783, respectively, all P > 0.05). (3) Multivariate Logistic regression analysis showed that NLR on day 7 was an independent risk factor for sepsis progression to CCI [odds ratio (OR) = 1.155, P = 0.005]. (4) ROC curve analysis showed that the area under the curve (AUC) of NLR predicting the sepsis progression to CCI on day 7 was 0.775, and the 95% confidence interval (95%CI) was 0.670-0.860, P < 0.01; when the cut-off value was 9.25, the sensitivity was 69.57% and the specificity was 80.56%. CONCLUSIONS: Dynamic monitoring of NLR is helpful to determine the progress of sepsis in elderly patients, NLR on day 7 has a certain predictive value for the occurrence of CCI.

[Nutrition support in the chronic critically ill patients].

Over the last decade, chronic critically ill (CCI) has emerged as an epidemic in intensive care unit (ICU) survivors worldwide. Advances in ICU technology and implementation of care bundles has significantly decreased early deaths of critically ill patients, and have allowed them to survive previously lethal multiple organ failure (MOF). However, more and more survivors leave persistent low grade organ dysfunctions, depend on continues organ support, need to stay in ICU, and become CCI patients. These patients experience a persistent immune dysregulation with persistent inflammation, immunosuppression, and catabolic syndrome. Therefore, malnutrition is an important feature of patients with CCI, and nutritional support is a crucial part of their treatment. The main strategies of nutritional support are as follows: providing sufficient calories and proteins with appropriate anabolic agents to promote anabolic metabolism, using immunomodulators to improve immune suppression and inflammatory responses, and supplementing micronutrients to enhance metabolic support. In this review, the nutritional assessment, calorie assessment, protein assessment and other nutrient supplementation (such as ß blocker, testosterone and oxandrolone, immunonutrition, vitamins) of CCI patients were reviewed, so as to provide reference for the treatment of CCI.

[A 180-day mortality predictive score based on frailty syndrome in elderly patients with sepsis: a Logistic regression analysis model].

OBJECTIVE: To establish a 180-day mortality predictive score based on frailty syndrome in elderly sepsis patients [elderly sepsis score (ESS)]. METHODS: A prospective study for sepsis patients aged 60 years and above who were admitted to a medical intensive care unit of the General Hospital of Southern Theatre Command from January 1st, 2018 to December 31st, 2018 was conducted. Univariate analysis was performed on 19 independent variables including gender, age, body mass index (BMI), tumor, charlson comorbidity index (CCI), activity of daily living (ADL), instrumental activity of daily living (IADL), mini-mental state examination (MMSE), geriatric depression scale (GDS), clinical frail scale (CFS), sequential organ failure assessment (SOFA), Glasgow coma scale (GCS), acute physiology and chronic health evaluation (APACHE II, APACHE IV), modified NUTRIC score (MNS), multiple drug resistance (MDR), mechanical ventilation (MV), continuous renal replacement therapy (CRRT) and palliative care. Continuous independent variables were converted into classified variables. Multivariate binary regression analysis of risk factors was conducted to screen independent risk factors which affecting 180-day mortality in elderly sepsis patients. Then a 180-daymortality predictive score was established, and the discrimination of the mortality of patients using CFS, SOFA, GCS, APACHE II, APACHE IV, MNS scores were compared. RESULTS: A total of 257 patients were enrolled, with a 180-day mortality of 60.7%. Univariate analysis showed that age, tumor, CCI, ADL, IADL, MMSE, CFS, SOFA, GCS, APACHE II, APACHE IV, MNS, MDR, MV, CRRT, palliative care were risk factors of 180-day mortality in elderly sepsis patients [age: odds ratio (OR) = 1.027, 95% confidence interval (95%CI) was 1.005-1.050, P = 0.018; tumor: OR =2.001, 95%CI was 1.022-3.920, P = 0.043; CCI: OR = 1.193, 95%CI was 1.064-1.339, P = 0.003; ADL: OR = 0.851, 95%CI was 0.772-0.940, P = 0.001; IADL: OR = 0.894, 95%CI was 0.826-0.967, P = 0.005; MMSE: OR = 0.962, 95%CI was 0.937-0.988, P = 0.004; CFS: OR = 1.303, 95%CI was 1.089-1.558, P = 0.004; SOFA: OR = 1.112, 95%CI was 1.038-1.191, P = 0.003; GCS: OR = 0.918, 95%CI was 0.863-0.977, P = 0.007; APACHE II: OR = 1.098, 95%CI was 1.053-1.145, P < 0.001; APACHE IV: OR = 1.032, 95%CI was 1.020-1.044, P < 0.001; MNS: OR = 1.315, 95%CI was 1.159-1.493, P < 0.001; MDR: OR = 2.029, 95%CI was 1.197-3.437, P = 0.009; MV: OR = 6.408, 95%CI was 3.480-11.798, P < 0.001, CRRT: OR = 2.744, 95%CI was 1.529-4.923, P = 0.001, palliative care: OR = 5.760, 95%CI was 2.177-15.245, P < 0.001]. By binary regression analysis, CFS stratification (OR = 1.934, 95%CI was 1.267-2.953, P = 0.002), MV (OR = 4.531, 95%CI was 2.376-8.644, P < 0.001), CRRT (OR = 2.471, 95%CI was 1.285-4.752, P = 0.007), palliative care (OR = 6.169, 95%CI was 2.173-17.515, P = 0.001) were independent risk factors of 180-day mortality in elderly patients with sepsis. The model of "ESS = 0.660×CFS stratification+1.511×MV+0.905×CRRT+1.820×palliative care" was established. Receiver operating characteristic curve (ROC curve) analysis showed that the area under the ROC curve (AUC) for predicting 180-day mortality by ESS was 0.785 (95%CI was 0.730-0.834, P < 0.001). When the best cut-off value was 2.2 points, its sensitivity was 78.9%, specificity was 70.3%, the positive predictive value was 80.4%, and the negative predictive value was 68.3%. Simplified ESS was defined as "0.5×CFS stratification+1.5×MV+1×CRRT+2×palliative care". ROC curve analysis showed that AUC for predicting 180-day mortality by simplified ESS was 0.784 (95%CI was 0.729-0.833, P < 0.001). When the best cut-off value was 2.0 points, sensitivity was 76.9%, specificity was 70.3%, the positive predictive value was 80.0%, and the negative predictive value was 66.4%. Compared with CFS, SOFA, GCS, APACHE II, APACHE IV and MNS, ESS had a significant difference in discriminating 180-day mortality in elderly patients with sepsis (AUC was 0.785 vs. 0.607, 0.607, 0.600, 0.664, 0.702, 0.657, 95%CI: 0.730-0.734 vs. 0.537-0.678, 0.537-0.677, 0.529-0.671, 0.598-0.730, 0.638-0.766, 0.590-0.725, all P < 0.05). CONCLUSIONS: CFS, MV, CRRT, and palliative care are independent risk factors of 180-day mortality in elderly patients with sepsis. We established ESS based on these risk factors. The ESS model has good discrimination and can be used as a reference and assessment tool for prediction and treatment guidance in elderly patients with sepsis.

Swertianolin ameliorates immune dysfunction in sepsis via blocking the immunosuppressive function of myeloid- derived suppressor cells.

In this study, we studied the long-term proliferation trajectory of myeloid-derived suppressor cells (MDSCs) in murine sepsis model and investigated whether swertianolin could modulate the immunosuppressive function of MDSCs. A murine sepsis model was established by cecal ligation and perforation (CLP), according to the Minimum Quality Threshold in Pre-Clinical Sepsis Studies (MQTiPSS) guidelines. The bone marrow and spleen of the mice were collected at 24 h, 72 h, 7 and 15 d after sepsis induction. The proportions of monocytic-MDSCs (M-MDSCs; CD11b+LY6G-LY6Chi) and granulocytic-MDSCs (G-MDSC, CD11b+ Ly6G+ Ly6Clow) were analyzed by flow cytometry. Then, we have investigated whether swertianolin could modulate the immunosuppressive function of MDSCs in in vitro experiments. G-MDSCs and M-MDSCs increased acutely after sepsis with high levels sustained over a long period of time. G-MDSCs were the main subtype identified in the murine model of sepsis with polymicrobial peritonitis. Furthermore, it was found that swertianolin reduced significantly interleukin-10 (IL-10), nitric oxide (NO), reactive oxygen species (ROS), and arginase production in MDSCs, while reducing MDSC proliferation and promoting MDSC differentiation into dendritic cells. Swertianolin also improved T-cell activity by blocking the immunosuppressive effect of MDSCs. Both subsets of MDSCs significantly increased in the bone marrow and spleen of the mice with sepsis, with G-MDSCs being the main subtype identified. Swertianolin effectively regulated the functions of MDSCs and reduced immune suppression.

[Role of assessment and monitoring of human leucocyte antigen-DR on CD14+ monocyte in postoperative infection in patients after orthotopic liver transplantation].

OBJECTIVE: To investigate the changes in expression level of human leucocyte antigen-DR (HLA-DR) on CD14(+) monocyte (CD14(+)/HLA-DR) in the patients after orthotopic liver transplantation, and its role in monitoring postoperative infection. METHODS: Sixty-three patients with liver transplantation were divided into three groups, non-infection group with 47 cases, infection group with 10 cases and septic shock group with 6 cases [according to the definition of septic shock of American College of Chest Physicians/Society for Critical Care Medicine (ACCP/SCCM)]. CD14(+)/HLA-DR expression ratio was assessed with flow cytometer, and its clinical implication was evaluated by receiver operating characteristic (ROC) curve assay. RESULTS: CD14(+)/HLA-DR expression ratio in infection group [(29.6+/-7.2)%] and septic shock group [(16.3+/-10.5)%] were significantly lower than that in non-infection group [(62.3+/-18.3)%, both P<0.01], but no significant difference of CD14(+)/HLA-DR expression ratio was found between infection group and septic shock group (P=0.128). Total area under ROC curve of CD14(+)/HLA-DR expression ratio for the infection was 0.965, its sensitivity and specificity at 36.35% cut off were 100.0% and 93.6%, respectively. Total area under ROC curve of CD14(+)/HLA-DR expression ratio to predict septic shock was 0.968, its sensitivity and specificity at 31.97% cut off were 100.0% and 87.7%, respectively. Comparing the change of CD14(+)/HLA-DR expression, it was lower in the infection group and septic shock group (P<0.05 and P<0.01), and the expression rate was lowest during period of serious infection in the two groups [infection group: (29.6+/-7.2)%, septic shock group: (16.3+/-0.5)%, all P<0.01]. CONCLUSION: For the patients with possible infection after liver transplantation, sequential assessment of CD14(+)/HLA-DR expression ratio would be a good marker for the judgment of patient's conditions and outcome. CD14(+)/HLA-DR expression ratio below 36.35% could be used as the prewarning value for the diagnosis of postoperative infection, and 31.97% could be used as the critical value for the diagnosis of septic shock.

Downregulation of lncRNA NEAT1 Ameliorates LPS-Induced Inflammatory Responses by Promoting Macrophage M2 Polarization via miR-125a-5p/TRAF6/TAK1 Axis.

The lncRNA nuclear enriched abundant transcript 1 (NEAT1) promotes sepsis-inflammatory responses and acute kidney injury (AKI), but little known about the underlying mechanisms. This study aims to investigate the roles of NEAT1 in regulating macrophage polarization and its potential for alleviating inflammatory responses during sepsis pathogenesis. Mouse RAW264.7 macrophages were treated with lipopolysaccharide (LPS) as a cellular inflammatory model. NEAT1 shRNA, miR-125a-5p mimics, and TRAF6-overexpressing vector were used to transfect RAW264.7 cells. NEAT1, miR-125a-5p, and mRNA levels of functional genes were detected by quantitative RT-PCR. Protein abundances were analyzed by western blotting. Macrophage polarization was evaluated by flow cytometry. The bindings of miR-125a-5p with NEAT1 or TRAF6 gene were validated by dual luciferase reporter assay. LPS treatment promoted NEAT1 and suppressed miR-125a-5p expression in mouse macrophage cells. NEAT1 silencing by shRNAs promoted macrophage M2 polarization under LPS treatment, which upregulated miR-125a-5p expression, repressed TRAF6 expression and TAK1 protein phosphorylation in macrophages. These cellular and molecular changes induced by NEAT1 shRNAs were abrogated by miR-125a-5p inhibitors. Moreover, miR-125a-5p mimics suppressed TRAF6 expression and TAK1 protein phosphorylation in LPS-treated macrophages, thus causing macrophage M2 polarization under LPS treatment. TRAF6 overexpression abrogated the miR-125a-5p mimics-induced macrophage M2 polarization. miR-125a-5p could directly bind to NEAT1 or TRAF6 gene in macrophages. lncRNA NEAT1 knockdown ameliorates LPS-induced inflammation by promoting macrophage M2 polarization via miR-125a-5p/TRAF6/TAK1 axis.

[Effect of C5a on expression of thrombomodulin in endothelial cells in vitro].

OBJECTIVE: To investigate the effect of C5a on the expression of thrombomodulin (TM) in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs cultured in vitro were stimulated with C5a for 8, 12, 16, 20 hours in a concentration of 200 microg/L, and also with different concentrations of 100, 200, 300 microg/L for 12 hours respectively. The levels of both mRNA and protein expression of TM was detected by real-time polymerase chain reaction (PCR) and Western blotting respectively, and the dose and time dependent effects of C5a on the expression of TM were evaluated. RESULTS: C5a down-regulated the TM expression at both protein and mRNA level. The down-regulation was time-dependent [(93.11+/- 1.57) x10(-2) , (71.05+/-3.39)x10(-2), (65.48+/-4.28)x10(-2), (62.69+/-4.03)x10(-2)at protein level and (301.71+/-80.40)x10(-6), (38.29+/-20.24)x10(-6), (8.82+/-2.66)x10(-6), (7.05+/-0.80)x10(-6) at mRNA level, all P<0.05] and dose-dependent [(113.25+/-3.97)x10(-2), (80.18+/-2.56)x10(-2), (73.22+/- 4.36)x10(-2) at protein level and (401.77+/-20.46)x10(-6), (31.12+/-3.51)x10(-6), (18.19+/-1.46)x10(-6) at mRNA level , all P<0.05]. When concentration of C5a at 300 microg/L was used to stimulate HUVECs for longer than 12 hours, the lowering of TM at protein level was slowed down obviously. And concentration of C5a at 200 microg/L was used to stimulate HUVECs for 12 hours, the lowering of TM at mRNA level was slowed down obviously. CONCLUSION: C5a can depress the gene expression of TM, and then affect the protein's translation. By this means, C5a can lead to hypercoagulability and inflammatory injuries in sepsis.

Two RING-Finger Ubiquitin E3 Ligases Regulate the Degradation of SPX4, An Internal Phosphate Sensor, for Phosphate Homeostasis and Signaling in Rice.

SPX-domain-containing proteins (SPXs) play an important role in inorganic phosphate (Pi) sensing, signaling, and transport in eukaryotes. In plants, SPXs are known to integrate cellular Pi status and negatively regulate the activity of Pi central regulators, the PHOSPATE STARVATION RESPONSE proteins (PHRs). The stability of SPXs, such as SPX4, is reduced under Pi-deficient conditions. However, the mechanisms by which SPXs are degraded remain unclear. In this study, using a yeast-two-hybrid screen we identified two RING-finger ubiquitin E3 ligases regulating SPX4 degradation, designated SDEL1 and SDEL2, which were post-transcriptionally induced by Pi starvation. We found that both SDELs were located in the nucleus and cytoplasm, had ubiquitin E3 ligase activity, and directly ubiquitinated the K213 and K299 lysine residues in SPX4 to regulate its stability. Furthermore, we found that PHR2, a Pi central regulator in rice, could compete with SDELs by interacting with SPX4 under Pi-sufficient conditions, which protected SPX4 from ubiquitination and degradation. Consistent with the biochemical function of SDEL1 and SDEL2, overexpression of SDEL1 or SDEL2 resulted in Pi overaccumulation and induced Pi-starvation signaling even under Pi-sufficient conditions. Conversely, their loss-of-function mutants displayed decreased Pi accumulation and reduced Pi-starvation signaling. Collectively, our study revealed that SDEL1 and SDEL2 facilitate the degradation of SPX4 to modulate PHR2 activity and regulate Pi homeostasis and Pi signaling in response to external Pi availability in rice.

[Effect of mechanical ventilation with different tidal volume on small intestine in dogs suffering from acute lung injury].

OBJECTIVE: To compare the effect of different tidal volume (VT) on intestinal tissue in oleic acid-induced acute lung injury (ALI) dogs undergoing mechanical ventilation (MV). METHODS: ALI was induced with oleic acid in dogs. While all of them were undergoing MV, they were randomized into two groups: low VT group (n=6), with VT 6 ml/kg, positive end-expiratory pressure (PEEP) 10 cm H2O (1 cm H2O= 0.098 kPa), and large VT group (n=6), with VT 20 ml/kg, PEEP 10 cm H2O. MV with different VT was maintained for 6 hours. After 6 hours, dogs were sacrificed by exsanguination. Pathological changes in small bowel tissues were observed with hematoxylin and eosin (HE) staining, and cellular apoptosis detected with terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling (TUNEL) in situ apoptosis determination. RESULTS: After 6-hour MV, the degree of flatulence was severe in the large VT group, but no flatulence was observed in the low VT group. Bowel injury score was lower in the low VT group than in large VT group [(3.17+/-0.75) scores vs. (2.00+/-0.89) scores]. There was significant difference between two groups (P<0.01). Apoptosis-positive cells were rare in the small bowel tissues in both groups. CONCLUSION: MV with large VT can induce bowel dysfunction, in contrast low VT MV does not produce small bowel dysfunction.

[Effect of immune function on prognosis of patients with sepsis].

OBJECTIVE: To investigate the influence of immune function and its changes on the prognosis of patients with sepsis. METHODS: 393 patients who met the diagnostic criteria of Sepsis-3 admitted to General Hospital of Southern War Zone of PLA from April 2003 to April 2017 were enrolled. Clinical data were collected and analyzed retrospectively. According to the initial immune status, patients with more than 4 days course of disease were divided into the initial immune suppression group (219 cases) and the initial immune function normal group (174 cases). According to the changes of immune function, patients with more than 7 days course of disease were divided into persistent inhibition group (113 cases), persistent normal group (96 cases), first normal inhibition group (22 cases) and first inhibited normal group (59 cases). In addition, the patients were divided into the elderly group (≥ 65 years old) and the young group (< 65 years old). Acute physiology and chronic health evaluation II (APACHE II), sequential organ failure assessment (SOFA), procalcitonin (PCT), C-reactive protein (CRP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), serum creatinine (SCr) within 24 hours after diagnosis of sepsis, whether respiratory failure and circulatory failure occur, hormone, immunomodulatory drugs and high-volume hemofiltration treatment within 28 days, the absolute value of lymphocyte counts for 4 consecutive days after diagnosis and 4 consecutive days before the end point event (death or survival within 28 days or more than 28 days) were collected and compared between each group. RESULTS: Among 393 sepsis patients, 174 cases had normal initial immune function, of whom 85 cases were older than 65 years old; 219 cases had depression of initial immune function, of whom 118 cases were older than 65 years old. Compared with the initial immune function normal group, the levels of PCT, CRP, ALT, AST and SCr in the initial immunosuppressive group were significantly increased [PCT (µg/L): 9.32 (2.13, 34.01) vs. 4.28 (1.02, 19.02), CRP (mg/L): 89.00 (26.00, 142.00) vs. 65.25 (19.88, 119.04), ALT (mmol/L): 39.0 (39.0, 99.0) vs. 27.0 (16.2, 73.0), AST (mmol/L): 55.0 (31.0, 148.0) vs. 39.0 (23.0, 100.8), SCr (µmol/L): 132.00 (74.75, 245.00) vs. 100.25 (61.00, 182.54)], the mean absolute value of lymphocyte counts for 4 consecutive days was significantly decreased [0.615 (0.380, 0.810) vs. 1.442 (1.217, 1.742)], SOFA and APACHE II were significantly increased (SOFA: 9.25±4.19 vs. 6.87±4.66, APACHE II: 22.27±8.96 vs. 18.25±9.47), the incidence of circulatory failure (66.2% vs. 50.0%), the incidence of respiratory failure (87.7% vs. 69.0%) and 28-day mortality (65.3% vs. 33.9%) were significantly increased, with statistically significant differences (all P < 0.05). When combined with immunosuppression, there was no significant difference in 28-day mortality between the elderly group and the young group (26.3% vs. 15.8%, P > 0.05); when the immune function was normal, the 28-day mortality of the elderly group was significantly higher than that of the young group (48.2% vs. 20.2%, P < 0.01). The 28-day mortality of the persistent inhibition group and the first normal inhibition group were significantly higher than those of the persistent normal group and the first inhibition normal group [83.2% (94/113), 81.8% (18/22) vs. 26.0% (25/96), 40.7% (24/59), all P < 0.05]. The incidence of immunosuppression in elderly patients [33.3% (14/42) vs. 10.5% (8/76)] and the incidence of persistent immunosuppression [77.0% (67/87) vs. 54.1% (46/85)] were higher than those in young patients (all P < 0.01). CONCLUSIONS: Immune function is closely related to the prognosis of sepsis patients. Elderly patients with sepsis are more likely to have immunosuppression or persistent immunosuppression than young patients, and the prognosis is worse.