Identification of qualitative characteristics of immunosuppression in sepsis based on immune-related genes and immune infiltration features.

Objective: Sepsis is linked to high morbidity and mortality rates. Consequently, early diagnosis is crucial for proper treatment, reducing hospitalization, and mortality rates. Additionally, over one-fifth of sepsis patients still face a risk of death. Hence, early diagnosis, and effective treatment play pivotal roles in enhancing the prognosis of patients with sepsis. Method: The study analyzed whole blood data obtained from patients with sepsis and control samples sourced from three datasets (GSE57065, GSE69528, and GSE28750). Commonly dysregulated immune-related genes (IRGs) among these three datasets were identified. The differential characteristics of these common IRGs in the sepsis and control samples were assessed using the REO-based algorithm. Based on these differential characteristics, samples from eight Gene Expression Omnibus (GEO) databases (GSE57065, GSE69528, GSE28750, GSE65682, GSE69063, GSE95233, GSE131761, and GSE154918), along with three ArrayExpress databases (E-MTAB-4421, E-MTAB-4451, and E-MTAB-7581), were categorized and scored. The effectiveness of these differential characteristics in distinguishing sepsis samples from control samples was evaluated using the AUC value derived from the receiver operating characteristic curve (ROC) curve. Furthermore, the expression of IRGs was validated in peripheral blood samples obtained from patients with sepsis through qRT-PCR. Results: Among the three training datasets, a total of 84 common dysregulated immune-related genes (IRGs) were identified. Utilizing a within-sample relative expression ordering (REOs)-based algorithm to analyze these common IRGs, differential characteristics were observed in three reverse stable pairs (ELANE-RORA, IL18RAP-CD247, and IL1R1-CD28). In the eight GEO datasets, the expression of ELANE, IL18RAP, and IL1R1 demonstrated significant upregulation, while RORA, CD247, and CD28 expression exhibited notable downregulation during sepsis. These three pairs of immune-related marker genes displayed accuracies of 95.89% and 97.99% in distinguishing sepsis samples among the eight GEO datasets and the three independent ArrayExpress datasets, respectively. The area under the receiver operating characteristic curve ranged from 0.81 to 1.0. Additionally, among these three immune-related marker gene pairs, mRNA expression levels of ELANE and IL1R1 were upregulated, whereas the levels of CD247 and CD28 mRNA were downregulated in blood samples from patients with sepsis compared to normal controls. Conclusion: These three immune-related marker gene pairs exhibit high predictive performance for blood samples from patients with sepsis. They hold potential as valuable auxiliary clinical blood screening tools for sepsis.

DLK1 overexpression improves sepsis-induced cardiac dysfunction and fibrosis in mice through the TGF-ß1/Smad3 signaling pathway and MMPs.

Sepsis is a serious inflammatory disease caused by bacterial infection. Cardiovascular dysfunction and remodeling are serious complications of sepsis, which can significantly affect sepsis patients' mortality. Delta-like homologue 1 (DLK1) has been reported could inhibit cardiac myofibroblast differentiation. However, the function of DLK1 in sepsis is unknown. In the present study, the DLK1 expression was first identified based on the online dataset GSE79962 analysis and cecal ligation and puncture (CLP)-induced sepsis mouse model. DLK1 expression was significantly reduced in septic heart tissues. In septic mouse heart, CLP operation decreased the fractional shortening (EF) (%) and ejection fraction (FS) (%) and caused significant edema, disordered myofilament arrangement, and degradation and necrosis in myocardial cells; CLP operation also increased collagen deposition and elevated the protein levels of fibrotic markers (α-SMA and F-actin). DLK1 overexpression in septic mice could effectively increase EF (%) and FS (%), attenuate CLP-caused ECM degradation and deposition and partially inhibit the CLP-induced TGF-ß1/Smad signaling activation. In conclusion, DLK1 expression was poorly expressed in the CLP-induced septic mouse heart. DLK1 overexpression partially alleviated sepsis-induced cardiac dysfunction and fibrosis, with the involvement of the TGF-ß1/Smad3 signaling pathway and MMPs.

KLF13 overexpression protects sepsis-induced myocardial injury and LPS-induced inflammation and apoptosis.

Sepsis remains a worldwide public health problem. This study aims to explore the role and mechanism of transcriptional factors (TFs) in sepsis-induced myocardial injury. Firstly, TF KLF13 was selected to explore its role in sepsis-induced myocardial injury. The caecal ligation and puncture (CLP) -induced sepsis mouse model was established and the septic mice were examined using standard histopathological methods. KLF13 expression was detected in the septic mouse heart and was also seen in a lipoploysaccharide (LPS) -induced cellular inflammation model. To explore this further both pro-apoptotic cleaved-caspase3/caspase3 and Bax levels and anti-apoptotic Bcl2 levels were examined, also in both models, In addition inflammatory cytokine (IL-1ß, TNF-α, IL-8 and MCP-1) production and IκB-α protein level and p65 phosphorylation were examined in both septic mice and LPS-induced cells. Thus three parameters - cardiomyocyte apoptosis, inflammatory response and NF-κB pathway activation were evaluated under similar conditions. The septic mice showed significant oedema, disordered myofilament arrangement and degradation and necrosis to varying degrees in the myocardial cells. KLF13 was downregulated in both the septic mouse heart and the LPS-induced cellular inflammation model. Furthermore, both models showed abnormally increased cardiomyocyte apoptosis (increased cleaved-caspase3/caspase and Bax protein levels and decreased Bcl2 level), elevated inflammation (increased production of inflammatory cytokines) and the activated NF-κB pathway (increased p65 phosphorylation and decreased IκB-α protein level). KLF13 overexpression notably ameliorated sepsis-induced myocardial injury in vivo and in vitro. KLF13 overexpression protected against sepsis-induced myocardial injury and LPS-induced cellular inflammation and apoptosis via inhibiting the inflammatory pathways (especially NF-κB signalling) and cardiomyocyte apoptosis.

Osteocalcin Induces Proliferation via Positive Activation of the PI3K/Akt, P38 MAPK Pathways and Promotes Differentiation Through Activation of the GPRC6A-ERK1/2 Pathway in C2C12 Myoblast Cells.

BACKGROUND/AIMS: Sarcopenia is characterized by an age-related decline in skeletal muscle plus low muscle strength and/or physical performance. Despite the clinical significance of sarcopenia, the molecular pathways underlying sarcopenia remain elusive. The recent demonstration that undercarboxylated osteocalcin (ucOC) favours muscle function related to insulin sensitivity and glucose metabolism raises the question of whether this hormone may also regulate muscle mass. The present study explored the promotive effects of ucOC in proliferation and differentiation processes of C2C12 myoblasts as well as the possible signalling pathways involved. METHODS: The effects of exogenous ucOC on C2C12 myoblasts proliferation were assessed using CCK8 and immunohistological staining assays. C2C12 cells were pretreated with PI3K/Akt or P38 MAPK inhibitors to investigate the possible involvement of the PI3K/Akt and P38 MAPK pathways in proliferation. The levels of Akt, phosphorylated-Akt (p-Akt), P38, and phosphorylated-P38 (p-P38) were measured by Western Blotting. The effects of ucOC on myoblast differentiation were quantified by morphological analysis. A silencing experiment was conducted in which the expression of GPRC6A in C2C12 myoblasts was modified. The expression of GPRC6A, myosin heavy chain (MyHC) and the related ERK1/2 signalling pathway in C2C12 myoblasts were monitored by qRT-PCR and Western Blotting. RESULTS: We showed that treatment with exogenous ucOC stimulated the priming of C2C12 myoblasts proliferation. Inhibition of Akt phosphorylation by wortmannin or inhibition of P38 MAPK phosphorylation by SB203580 decreased C2C12 cell proliferation. Wortmannin also reduced P38 MAPK phosphorylation, whereas SB203580 did not affect Akt activation. Furthermore, ucOC promoted C2C12 myoblast differentiation. Inhibition of ERK1/2 phosphorylation with U0126 decreased C2C12 cell differentiation. Finally, GPRC6A expression was substantially increased after ucOC treatment of C2C12 cells. GPRC6A silencing inhibited Akt, P38 MAPK phosphorylation in C2C12 cells, and ERK1/2 phosphorylation in C2C12 myotubes; GPRC6A silencing also decreased cell proliferation, decreased cell differentiation, and downregulated MyHC expression. CONCLUSIONS: The present data suggest that ucOC induces myoblast proliferation via sequential activation of the PI3K/Akt and p38 MAPK pathways in C2C12 myoblast cells. Moreover, ucOC enhances myogenic differentiation via a mechanism involving GPRC6A-ERK1/2 signalling.

Effectiveness of a PRECEDE-based education intervention on quality of life in elderly patients with chronic heart failure.

BACKGROUND: One of the most important challenges in public health is to improve the quality of life in patients with chronic heart failure (CHF). Depression, self-care capacity, and quality of life interact each other in these patients. It's difficult to treat with general education programs and conventional therapy. PRECEDE model is a comprehensive and exclusive theory-based education programs. Its effectiveness for reducing depression and increasing quality of life has been demonstrated in patients with coronary artery bypass grafting, type 2 diabetes, and the elderly. It has not been used in elderly patients with CHF. Thus, this study aims to investigate the effects of this model on self-care behaviors, depression, and quality of life in these patients. METHODS: Patients who met the inclusion criteria were randomly assigned to the intervention or control group. All the patients received conventional medical care. The patients in the intervention group also received 9 sessions of education intervention based on the PRECEDE model and then followed up for 3 months after the intervention. Data were collected before and 3 months after the intervention using 4 questionnaires, namely a PRECEDE-based questionnaire to evaluate predisposing, reinforcing, and enabling factors; the 9-item European Heart Failure Self-care Behavior Scale (EHFScBS-9); the 9-item Personal Health Questionnaire (PHQ-9); and the Minnesota Living with Heart Failure Questionnaire (MLHFQ). RESULTS: No significant differences were found in the mean scores for the predisposing, enabling, and reinforcing factors, and the mean total scores in EHFScBS-9, PHQ-9, and MLHFQ before the intervention between the intervention and control groups. After the intervention, the scores for the predisposing, reinforcing, and enabling factors increased significantly, and the mean total scores in EHFScBS-9, PHQ-9, and MLHFQ decreased significantly in the intervention group. In addition, these scores significantly differed from those of the control group. Furthermore, the MLHFQ score significantly correlated with the EHFScBS-9 and PHQ-9 scores. CONCLUSION: This study demonstrates a trend that PRECEDE model of health education promotion is effective in relieving depression symptoms, enhancing self-monitoring, and improving the quality of life of elderly patients with CHF. TRIAL REGISTRATION: Trial registration number: ChiCTR-IOR-17012779 ; Trial registry: Chinese Clinical Trial Registry; Date registered: 22 Sep 2017; Retrospectively registered.

MicroRNA-142-3p inhibits hypoxia/reoxygenation­induced apoptosis and fibrosis of cardiomyocytes by targeting high mobility group box 1.

Myocardial ischemia/reperfusion (I/R) injury may cause the apoptosis of cardiomyocytes as well as cardiac fibrosis, which is characterized as the transdifferentiation of fibroblasts to myofibroblasts and collagen deposition. MicroRNAs (miRNAs or miRs) have been demonstrated to be involved in myocardial I/R injury. However, the underlying molecular mechanism remains largely unclear. In the present study, mouse cardiomyocyte M6200 cells were treated with hypoxia/reoxygenation (H/R). Our data indicated that H/R treatment led to cell apoptosis, the increased expression of fibrosis­related proteins, namely collagen I, II, III, and fibronectin, as well as the downregulation of miR-142-3p in M6200 cells. Overexpression of miR-142-3p suppressed the H/R-induced apoptosis and fibrosis of M6200 cells. Bioinformatics analysis and a Dual­Luciferase reporter assay further identified high mobility group box 1 (HMGB1) as a direct target gene of miR-142-3p, and miR-142-3p negatively regulated the protein level of HMGB1 in M6200 cells. Furthermore, knockdown of HMGB1 enhanced cell proliferation whereas it inhibited the apoptosis and fibrosis of M6200 cells. In addition, TGF-ß1/Smad3 signaling was suggested to be involved in the miR-142-3p/HMGB1-mediated apoptosis and fibrosis of M6200 cells treated with H/R. Taken together, the findings of the present study demonstrate that miR-142-3p inhibits H/R-induced apoptosis and fibrosis of cardiomyocytes, partly at least, by the direct inhibition of HMGB1 expression. Therefore, these findings have increased our understanding of the pathogenesis of H/R-induced myocardial injury.

[Correlation between resting heart rate and blood glucose level in elderly patients with coronary heart disease and diabetes mellitus].

OBJECTIVE: To explore the correlation between resting heart rate (RHR) and blood glucose level in elderly patients with coronary heart disease (CHD) complicated by diabetes mellitus. METHODS: Between April and July, 2011, a total of 1336 outpatients over 60 years of age recruited from 165 hospitals were asked to complete a questionnaire and received blood glucose and RHR examination. According to baseline RHR, the patients were divided into 3 groups with HRH <70 min-1 (group I, 372 cases), between 70 and 79 min(-1) (group II, 533 cases), and ≥80 min(-1) (group III, 431cases) for analysis of the relationships of RHR with blood glucose control rate. RESULTS: HbA1c levels in the total, male and female patients differed significantly among the 3 groups (F=15.436, 15.436, and 24.270, respectively, P<0.05), and increased in the order from group I to group III. Blood glucose control rate in the total, male and female patients also differed significantly among the 3 groups (χ(2)=13.471, 6.752, and 6.522, respectively, P<0.05), and was significantly lower in group III than in group I (P<0.05). RHR was found to positively correlate with FPG, 2 hPG and HbA1c by Pearson correlation analysis (r=0.058, 0.085, and 0.058, respectively; P<0.05) and multiple linear regression analysis (ß=0.075, 0.075, and 0.018, respectively; P<0.05). Multivariable logistic regression equation showed that compared with patients with RHR <70 min-1, the total, male and female patients with RHR ≥80 min(-1) had OR values of blood glucose control failure of 1.99 (95% CI: 1.23-2.37, P<0.05), 1.81 (95% CI: 1.17-2.77, P<0.05), and 2.18 (95% CI: 1.12-3.74, P<0.05), respectively. CONCLUSION: RHR in elderly CHD patients with MD is positively correlated with their blood glucose level, and an increased RHR is associated with an increased risk of poor blood glucose control. Rigorous RHR control in such high-risk patients may prove beneficial for both blood glucose control and secondary prevention of CHD.

[Current status of blood pressure control rate and antihypertensive drug use in the elderly coronary heart disease patients with diabetes mellitus and hypertension].

OBJECTIVE: To investigate the current status of blood pressure control rate and the use of antihypertensive drugs in elderly patients with coronary heart disease, diabetes mellitus and hypertension. METHODS: The elderly coronary heart disease patients with diabetes mellitus and hypertension (≥ 60 years old) were recruited from 165 hospitals in 21 provinces or cities across China from April to July 2011 in this multicenter, non-intervention and cross-sectional survey. The current status of blood pressure control rate in different antihypertensive target value, methods for application in antihypertensive drugs and standardized treatment recommended by guideline were investigated in the survey. RESULTS: 1 379 cases were eventually selected from the total 7 962 elderly patients (accounted for 17.3%). (1) The blood pressure control rate was 17.1% for antihypertensive target value (<130/80 mmHg); the control rate for 140/80 mmHg was 27.5%; the control rate for 140/90 mmHg was 39.6%; the success rate for 150/90 mmHg were 51.7%; control rate of elderly patient (≥ 70 years old) gradually increased with increasing of age; success rate of elderly patient (≥ 85 years old) was the highest, whereas control rate of elderly patient aged 71-71 years old was lowest; success rate for male patients was close to female patients, and success rate for men were slightly higher than those in women; (2) 1 347 cases had clear medication history (32 cases were missed) in the survey, 1 317 effective cases received antihypertensive therapy (effective rate was 97.8%, 1 317/1 347); the more commonly used drugs were angiotensin receptor blockers (ARB) /angiotensin converting enzyme inhibitors (ACEI) (usage rate was 76.8%), followed by dihydropyridine calcium channel blockers (CCB) (65.5%), ß-blockers (usage rate was 44.6%), thiazide diuretics (26.3%) respectively; (3) combinations of two drugs was the most common way in antihypertensive medication (accounted for 41.2%), three drugs or more was 28.9%, and single drug was 23.9%; CCB was the commonly used single drug (accounted for 8.8%); combinations of CCB and ARB were the most common way in combination of two drugs(11.7%), CCB combined with ARB and ß-blockers was frequently used in combination of three drugs or more (9.2%); (4) 987 cases received standardized treatment recommended by guideline (accounted for 76.6%); the percentage of standardized usage in combinations of two drugs was 71.9%, the percentage of standard usage in combinations of three drugs or more was 66.1%. CONCLUSION: The percentage of antihypertensive therapy is high, however, the overall blood pressure control rate is low. ACEI /ARB are the major drugs in antihypertensive medication, diuretic drugs are now rarely used; combined medication is the common method of antihypertensive therapy; the consciousness following the guidelines has improved, but still need to be strengthened.

Vitamin D Binding Protein Affects the Correlation of 25(OH)D and Frailty in the Older Men.

Vitamin D binding protein (DBP) may alter the biologic activity of 25-hydroxyvitamin D [25(OH)D]. The objective of our present study was to determine the joint effect of serum 25(OH)D and DBP on the risk of frailty. Five hundred sixteen male participants aged 70 years or older were recruited in Changsha city and its surrounding area in Hunan province of China. Frailty was defined as the presence of at least three of the five following criteria: weakness, low physical activity, slow walking speed, exhaustion, and weight loss. Multivariate linear regression analysis was performed to assess the relationship between 25(OH)D and DBP levels. Odds ratios (ORs) for frailty were evaluated across quartiles of 25(OH)D and DBP levels, adjusted age, education, and body mass index. The results showed that participants in the lowest quartile of 25(OH)D and the highest quartile of DBP levels, the lowest quartile of 25(OH)D and the lowest quartile of DBP levels, and those in the the lower quartile of 25(OH)D and lowest quartile of DBP levels had significantly higher OR of being frail compared with those in the highest quartile of 25(OH)D and lowest quartile of DBP, with OR of 3.18 (95% CI: 1.46-4.56, P < 0.05), 2.63 (95% CI: 1.31-3.68, P < 0.01), and 2.52 (95% CI: 1.22-3.52, P < 0.05), respectively. The results indicate that the joint effect of serum 25(OH)D and DBP levels is associated with the risk of frailty, and serum DBP levels affects 25(OH)D-frailty relationship in the older men.

Efficacy and safety of acarbose in the treatment of elderly patients with postprandial hypotension.

BACKGROUND: Postprandial hypotension (PPH) occurs frequently in elderly people and may lead to syncope, falls, dizziness, weakness, angina pectoris, and stroke. Some studies suggest that the magnitude of the postprandial fall in blood pressure (BP) is influenced by the rate at which glucose enters the small intestine. We hypothesized that acarbose (alpha-glucosidase inhibitor), a hypoglycemic agent that decreases the rate of glucose absorption in the small intestine, would attenuate PPH in the elderly, and would be safe in the treatment. METHODS: Forty-three elderly in-patients with PPH were recruited. All of them were in relatively stable conditions. They had semi-liquid standard meals without and with acarbose for the two following days: screening day and intervention day. Blood pressure and heart rate (HR) were recorded at baseline and every 15 minutes for 120 minutes using a non-invasive ambulatory blood pressure monitoring system during the study, and ejection fraction (EF) and fractional shortening (FS) were measured by two dimensional echocardiography. RESULTS: Compared with the screening day, the falls in systolic, diastolic and mean arterial blood pressure (SBP, DBP, MAP) (all P < 0.05) were significantly attenuated after taking acarbose during breakfast, so were MAP (P < 0.05) during lunch, DBP (P < 0.05) and MAP (P < 0.05) during supper. The change of HR was not statistically significant after taking acarbose in three meals. EF and FS were positively correlated with the relief rate. The effective power was 63%, and the incidence of adverse drug reaction (ADR) was 9%. CONCLUSION: Acarbose is effective and safe in the treatment of elderly patients with PPH.

[Cerebrospinal fluid levels of beta-amyloid protein 1-42 in Alzhemier disease].

OBJECTIVE: To study the role of beta-amyloid protein 1-42 (A beta 1-42) content in the cerebrospinal fluid (CSF) of Alzheimer disease (AD) patients. METHODS: A beta 1-42 levels were measured with the ELISA method in AD (n = 30), non-AD (NAD, n = 25) and non-dementia (ND, n = 21). RESULTS: The A beta 1-42 mean value for AD was (109.91 +/- 58.78) fmol.L-1. In ND, the A beta 1-42 mean value was (242.40 +/- 142.58) fmol.L-1. The mean value for AD was significantly lower than that of ND. In NAD, the A beta 1-42 mean value was (231.70 +/- 143.94) fmol.L-1, and it was not significantly different from the mean value for ND. The A beta 1-42 level was positively correlated with the severity of AD symptoms, but not with the duration. A beta 1-42 levels in CSF of AD were significantly lower than that of ND, and they decreased as the severity of disease increased. CONCLUSION: Cerebrospinal fluid beta-amyloid 1-42 analyses may be of value in the clinical diagnosis of Alzheimer's disease, especially in the earlier course of the disease, when drug therapy may have the greatest effect but clinical diagnosis is particularly difficult.