Biomarkers Of Disease Severity Research Articles

Interleukin-1β and Interleukin-10 Profiles and Ratio in Serum of COVID-19 Patients and Correlation with COVID-19 Severity: A Time Series Study.

Coronavirus disease-19 (COVID-19) can, in severe cases, lead to cytokine-release syndrome owing to an excessive immune response. The release of different cytokines aggravates disease severity. IL-1β is a pro-inflammatory cytokine, while IL-10 is an anti-inflammatory cytokine, and both are involved in the human immune response to infection. This study aimed to determine whether serum levels of IL-1β and IL-10 and the ratio of the two over time in patients with COVID-19 could facilitate early identification of disease severity. An analytical, observational time-series design was employed. Fifty participants were enrolled between May and October 2020 and were divided into two groups-non-severe (n = 20), and severe (n = 30). IL-1β and IL-10 were analyzed using BD cytometric bead array sets. Association of the IL-1β:IL-10 ratio with COVID-19 severity was analyzed using a Mann-Whitney test and Fisher's exact test. Optimal cut-off values to predict disease severity were determined by Youden's index. In non-severe and severe groups, the median serum levels of IL-1β decreased on day 3 (1.72 ng/mL and 2.10 ng/mL, respectively), then increased on day 6 (2.05 ng/mL and 3.31 ng/mL, respectively). However, the median of IL-10 increased on day 3 (1.88 ng/mL and 2.30 ng/mL, respectively) and day 6 (2.02 ng/mL and 2.39 ng/mL, respectively). There was no significant association between the IL-1β:IL-10 ratio and COVID-19 severity at any time-point (p>0.05). The cutoff value of serum IL-10 between the two groups on days 0, 3, and 6 was 1.09 pg/mL (sensitivity: 66.6%; PPV: 71.4%), 2.11 pg/mL (sensitivity: 67.7%; PPV: 50.0%), and 2.08 pg/mL (sensitivity: 78.6%; PPV: 70.9%), respectively. The IL-1β:IL-10 ratio was not correlated to COVID-19 severity. However, owing to its high sensitivity, IL-10 may be a potential biomarker for disease severity in severe COVID-19.

The distance distribution of human microRNAs in MirGeneDB database

MicroRNAs (miRNAs) are small single-stranded non-coding RNAs around 22 nucleotide lengths found in organisms, playing an important role in cell differentiation, development, gene regulation, and apoptosis. The distance of disease miRNA biomarkers has been used to explore the association between various diseases as well as the association between virus and disease in the literature. To date, there have been no studies on deriving the distribution of the pairwise distance of human miRNAs. As the pairwise distance of miRNA biomarkers might be a useful tool in studying the disease association, in this paper, the distance distributions of human miRNAs were derived such that they could be used to measure the closeness between miRNAs. Two distance models were used to calculate the pairwise distances of 567 Homo sapiens miRNA genes accessed from the MirGeneDB database. These miRNA pairwise distances were fitted by the normal distribution, gamma distribution, empirical cumulative distribution, and the kernel density estimation method. This is the first study to provide the distance distribution of human miRNAs. The similarity of miRNA biomarkers for several diseases was examined using the derived distributions.

Perivascular spaces as a marker of disease severity and neurodegeneration in patients with behavioral variant frontotemporal dementia.

Behavioural Variant Frontotemporal Dementia (bvFTD) is a rapidly progressing neurodegenerative proteinopathy. Perivascular spaces (PVS) form a part of the brain's glymphatic clearance system. When enlarged due to poor glymphatic clearance of toxic proteins, PVS become larger and more conspicuous on MRI. Therefore, enlarged PVS may be a useful biomarker of disease severity and progression in neurodegenerative proteinopathies such as bvFTD. This study aimed to determine the utility of PVS as a biomarker of disease progression in patients with bvFTD. Serial baseline and week 52 MRIs acquired from ten patients with bvFTD prospectively recruited and followed in a Phase 1b open label trial of sodium selenate for bvFTD were used in this study. An automated algorithm quantified PVS on MRI, which was visually inspected and validated by a member of the study team. The number and volume of PVS were extracted and mixed models used to assess the relationship between PVS burden and other measures of disease (cognition, carer burden scale, protein biomarkers). Additional exploratory analysis investigated PVS burden in patients who appeared to not progress over the 12 months of selenate treatment (i.e., "non-progressors"). Overall, PVS cluster number (ß = -3.27, CI [-7.80 - 1.27], p = 0.267) and PVS volume (ß = -36.8, CI [-84.9 - 11.3], p = 0.171) did not change over the paired MRI scans 12 months apart. There was association between cognition total composite scores and the PVS burden (PVS cluster ß = -0.802e-3, CI [9.45e - 3 - -6.60e - 3, p ≤ 0.001; PVS volume ß = -1.30e - 3, CI [-1.55e - 3 - -1.05e - 3], p ≤ 0.001), as well as between the change in the cognition total composite score and the change in PVS volume (ß = 4.36e - 3, CI [1.33e - 3 - 7.40e - 3], p = 0.046) over the trial period. There was a significant association between CSF t-tau and the number of PVS clusters (ß = 2.845, CI [0.630 - 5.06], p = 0.036). Additionally, there was a significant relationship between the change in CSF t-tau and the change in the number of PVS (ß = 1.54, CI [0.918 - 2.16], p < 0.001) and PVS volume (ß = 13.8, CI [6.37 - 21.1], p = 0.003) over the trial period. An association was found between the change in NfL and the change in PVS volume (ß = 1.40, CI [0.272 - 2.52], p = 0.045) over time. Within the "non-progressor" group (n = 7), there was a significant relationship between the change in the CSF total-tau (t-tau) levels and the change in the PVS burden (PVS cluster (ß = 1.46, CI [0.577 - 2.34], p = 0.014; PVS volume ß = 14.6, CI [3.86 - 25.4], p = 0.032) over the trial period. Additionally, there was evidence of a significant relationship between the change in NfL levels and the change in the PVS burden over time (PVS cluster ß = 0.296, CI [0.229 - 0.361], p ≤ 0.001; PVS volume ß = 3.67, CI [2.42 - 4.92], p = 0.002). Analysis of serial MRI scans 12 months apart in patients with bvFTD demonstrated a relationship between PVS burden and disease severity as measured by the total cognitive composite score and CSF t-tau. Further studies are needed to confirm PVS as a robust marker of neurodegeneration in proteinopathies.

Severe extra-glandular involvement and pleural effusions complicating primary Sjogren’s syndrome: a case report

BackgroundSjogren’s syndrome, an autoimmune disease of the exocrine glands, results in keratoconjunctivitis sicca, xerostomia, and dental caries. It is often overlooked, considered by clinicians to be a benign disease. However, it can cause life-threatening extra-glandular complications that affect multiple organ systems.Case presentationHere we present a 78-year-old Caucasian woman with a history of primary Sjogren’s syndrome (pSS) whose symptoms of keratoconjunctivitis sicca were managed managed conservatively. She was evaluated for sub-acute shortness of breath. Imaging showed severe bronchiectasis with features of lymphocytic interstitial pneumonia. She also had exudative bilateral pleural effusions and skin ulcers, likely vasculitic in origin. The workup was significant for anti-Ro antibody, pancytopenia, hypocomplementia, cryoglobulinemia and monoclonal gammopathy, all of which reflect disease severity. Although there was no evidence of malignancy, she developed B-cell non-Hodgkin lymphoma during follow-up.ConclusionsPrimary Sjogren’s syndrome can result in severe multi-organ disease. Pleural effusions are a rare complication of pSS, with only ten cases reported in the literature over the last 30 years, and tend to respond well to steroids. Prognostic biomarkers for disease severity include hypocomplementia, cryoglobulinemia, monoclonal gammopathy, and hypergammaglobulinemia. In this report we review the literature and the management of the disease.

Highly Sensitive, Flow Cytometry-Based Measurement of Intestinal Permeability in Models of Experimental Colitis

Increased intestinal permeability is seen in a variety of inflammatory conditions such as enteric infections and inflammatory bowel disease. Because barrier function can provide a key biomarker of disease severity, it often is assayed in animal models. A common methodology involves gavaging mice with fluorescein isothiocyanate-conjugated dextran (FITC-D), followed by cardiac puncture to assay plasma fluorescence on a spectrophotometer. Although the FITC-D method is relatively simple, its sensitivity is limited and enables only a single measurement because the test requires killing the subject. Herein, we describe a novel flow cytometry-based method of intestinal permeability measurement based on detection of orally gavaged ovalbumin (OVA) that leaks out of the gut. Our approach uses minute blood volumes collected from the tail vein, permitting repeated testing of the same subject at multiple time points. By comparing this assay against the gold standard FITC-D method, we show the expanded utility of our OVA assay in measuring intestinal permeability. We directly compared our OVA assay against the FITC-D assay by co-administering both probes orally to the same animals and subsequently using their respective methodologies to measure intestinal permeability by detecting probe levels in the plasma. Permeability was assessed in mice genetically deficient in intestinal mucus production or glycosylation. In addition, wild-type mice undergoing dextran sodium sulfate-induced colitis or infected by the enteric bacterial pathogen Citrobacter rodentium also were tested. The OVA assay showed very high efficacy in all animal models of intestinal barrier dysfunction tested. Besides identifying intestinal barrier dysfunction in mice with impaired mucin glycosylation, the assay also allowed for repeated tracking of intestinal permeability within the same animal over time, providing data that cannot be easily acquired with other currently applied methods. The OVA assay is a highly sensitive and effective method of measuring intestinal permeability in mouse models of barrier dysfunction and experimental colitis.

Dysregulation of glutamine/glutamate metabolism in COVID-19 patients: A metabolism study in African population and mini meta-analysis.

Coronavirus disease 2019 (COVID‐19) remains a serious global threat. The metabolic analysis had been successfully applied in the efforts to uncover the pathological mechanisms and biomarkers of disease severity. Here we performed a quasi‐targeted metabolomic analysis on 56 COVID‐19 patients from Sierra Leone in western Africa, revealing the metabolomic profiles and the association with disease severity, which was confirmed by the targeted metabolomic analysis of 19 pairs of COVID‐19 patients. A meta‐analysis was performed on published metabolic data of COVID‐19 to verify our findings. Of the 596 identified metabolites, 58 showed significant differences between severe and non‐severe groups. The pathway enrichment of these differential metabolites revealed glutamine and glutamate metabolism as the most significant metabolic pathway (Impact=0.5; ‐Log10P=1.959). Further targeted metabolic analysis revealed six metabolites with significant inter‐group differences, with glutamine/glutamate ratio significantly associated with severe disease, negatively correlated with 10 clinical parameters and positively correlated with SPO2 (rs=0.442, P=0.005). Mini meta‐analysis indicated elevated glutamate was related to increased risk of COVID‐19 infection (pooled OR=2.02; 95% CI: 1.17–3.50) and severe COVID‐19 (pooled OR=2.28; 95% CI: 1.14–4.56). In contrast, elevated glutamine related to decreased risk of infection and severe COVID‐19, the pooled OR were 0.30 (95% CI: 0.20–0.44), and 0.44 (95% CI: 0.19–0.98), respectively. Glutamine and glutamate metabolism are associated with COVID‐19 severity in multiple populations, which might confer potential therapeutic target of COVID‐19, especially for severe patients.This article is protected by copyright. All rights reserved.

Limited value of neutrophil-to-lymphocyte ratio and serum creatinine as point-of-care biomarkers of disease severity and infection mortality in patients hospitalized with COVID-19.

In hospitalized COVID-19, neutrophil-to-lymphocyte ratio (NLR) and serum creatinine is sometimes measured under assumption they predict disease severity and mortality. We determined the potential value of NLR and serum creatinine as predictors of disease severity and mortality in COVID-19. Prospective cohort study of COVID-19 patients admitted to premier COVID-19 treatment hospitals in Ethiopia. Predictive capability of biomarkers in progression and prognosis of COVID-19 was analyzed using receiver operating characteristics. Survival of COVID-19 patients with different biomarker levels was computed. Logistic regression assessed associations between disease severity and mortality on NLR and serum creatinine adjusted for odds ratio (AOR). The study enrolled 126 adults with severe (n = 68) or mild/moderate (n = 58) COVID-19, with median age 50 [interquartile range (IQR 20-86)]; 57.1% males. The NLR value was significantly higher in severe cases [6.68 (IQR 3.03-12.21)] compared to the mild/moderate [3.23 (IQR 2.09-5.39)], with the NLR value markedly associated with disease severity (p<0.001). Mortality was higher in severe cases [13 (19.1%)] compared to mild/moderate cases [2 (3.4%)] (p = 0.007). The NLR value was significantly higher in non-survivors [15.17 (IQR 5.13-22.5)] compared to survivors [4.26 (IQR 2.40-7.90)] (p = 0.002). Serum creatinine was significantly elevated in severe cases [34 (50%)] compared with mild/moderate [11 (19%)] (p<0.001). Disease severity [AOR 6.58, 95%CI (1.29-33.56), p = 0.023] and NLR [AOR 1.07, 95%CI (1.02-1.12), p = 0.004)] might be associated with death. NLR had a sensitivity and specificity of 69.1% and 60.3% as predictor of disease severity (cut-off >4.08), and 86.7% and 55.9% as prognostic marker of mortality (cut-off >4.63). In COVID-19, NLR is a biomarker with only modest accuracy for predicting disease severity and mortality. Still, patients with NLR >4.63 are more likely to die. Monitoring of this biomarker at the earliest stage of the disease may predict outcome. Additionally, high creatinine seems related to disease severity and mortality.

The fatal trajectory of pulmonary COVID-19 is driven by lobular ischemia and fibrotic remodelling.

SummaryBackgroundCOVID-19 is characterized by a heterogeneous clinical presentation, ranging from mild symptoms to severe courses of disease. 9–20% of hospitalized patients with severe lung disease die from COVID-19 and a substantial number of survivors develop long-COVID. Our objective was to provide comprehensive insights into the pathophysiology of severe COVID-19 and to identify liquid biomarkers for disease severity and therapy response.MethodsWe studied a total of 85 lungs (n = 31 COVID autopsy samples; n = 7 influenza A autopsy samples; n = 18 interstitial lung disease explants; n = 24 healthy controls) using the highest resolution Synchrotron radiation-based hierarchical phase-contrast tomography, scanning electron microscopy of microvascular corrosion casts, immunohistochemistry, matrix-assisted laser desorption ionization mass spectrometry imaging, and analysis of mRNA expression and biological pathways. Plasma samples from all disease groups were used for liquid biomarker determination using ELISA. The anatomic/molecular data were analyzed as a function of patients’ hospitalization time.FindingsThe observed patchy/mosaic appearance of COVID-19 in conventional lung imaging resulted from microvascular occlusion and secondary lobular ischemia. The length of hospitalization was associated with increased intussusceptive angiogenesis. This was associated with enhanced angiogenic, and fibrotic gene expression demonstrated by molecular profiling and metabolomic analysis. Increased plasma fibrosis markers correlated with their pulmonary tissue transcript levels and predicted disease severity. Plasma analysis confirmed distinct fibrosis biomarkers (TSP2, GDF15, IGFBP7, Pro-C3) that predicted the fatal trajectory in COVID-19.InterpretationPulmonary severe COVID-19 is a consequence of secondary lobular microischemia and fibrotic remodelling, resulting in a distinctive form of fibrotic interstitial lung disease that contributes to long-COVID.FundingThis project was made possible by a number of funders. The full list can be found within the Declaration of interests / Acknowledgements section at the end of the manuscript.

Pathogenic Roles of Heparan Sulfate and Its Use as a Biomarker in Mucopolysaccharidoses.

Heparan sulfate (HS) is an essential glycosaminoglycan (GAG) as a component of proteoglycans, which are present on the cell surface and in the extracellular matrix. HS-containing proteoglycans not only function as structural constituents of the basal lamina but also play versatile roles in various physiological processes, including cell signaling and organ development. Thus, inherited mutations of genes associated with the biosynthesis or degradation of HS can cause various diseases, particularly those involving the bones and central nervous system (CNS). Mucopolysaccharidoses (MPSs) are a group of lysosomal storage disorders involving GAG accumulation throughout the body caused by a deficiency of GAG-degrading enzymes. GAGs are stored differently in different types of MPSs. Particularly, HS deposition is observed in patients with MPS types I, II, III, and VII, all which involve progressive neuropathy with multiple CNS system symptoms. While therapies are available for certain symptoms in some types of MPSs, significant unmet medical needs remain, such as neurocognitive impairment. This review presents recent knowledge on the pathophysiological roles of HS focusing on the pathogenesis of MPSs. We also discuss the possible use and significance of HS as a biomarker for disease severity and therapeutic response in MPSs.

VP.25 Muscle MRI in female carriers of Duchenne muscular dystrophy

Muscle magnetic resonance imaging (mMRI) has been proved to be a reliable biomarker of disease severity and progression in patients with Duchenne and Becker muscular dystrophy (DMD/BMD). Although up to 20% of female carriers of DMD/BMD experienced symptoms, there is a paucity of data on mMRI in this patient group. Here, we report a case of a female DMD carrier who had mild clinical symptoms, yet mMRI revealed striking fatty infiltration. A 58-year-old female DMD carrier initially presented in 2020 with a 3-year history of slowly progressive symmetric pelvic girdle weakness since age 53. She was known to be a female DMD carrier at age 26 by linkage analysis, when her brother lost ambulation at age 10 and deceased at age 21 due to cardiopulmonary dysfunction. Subsequently, genetic testing of this patient and her mother both demonstrated deletion of exons 51-55 of the dystrophin gene. X inactivation study was not informative as the patient is homozygous for the polymorphic androgen receptor gene locus analyzed. On exam, the patient has normal muscle strength and a normal walking gait, but slight stepping to ascend steps, jump and hop. Brooke scale and functional Mobility Score were 1. Timed function test remained relatively stable over a time span of 22 months. mMRI of her pelvic and thigh muscles was performed to evaluate the progression of a dystrophic process. There was no significant edema in the musculature of the pelvis and thighs. However, there was striking fatty infiltration in multiple muscle groups, including glutei, adductors, hamstrings, as well as vastus lateralis and intermedius. This case of symptomatic female carrier suggests a possible clinical-radiological mismatch phenomenon in manifesting female carriers of DMD. Further study is indicated to understand the clinical implications of this observation.

IL-4, IL-10, CCL2 and TGF-β as potential biomarkers for severity in Plasmodium vivax malaria.

Cytokines and chemokines are immune response molecules that display diverse functions, such as inflammation and immune regulation. In Plasmodium vivax infections, the uncontrolled production of these molecules is thought to contribute to pathogenesis and has been proposed as a possible predictor for disease complications. The objective of this study was to evaluate the cytokine profile of P. vivax malaria patients with different clinical outcomes to identify possible immune biomarkers for severe P. vivax malaria. The study included patients with non-severe (n = 56), or severe (n = 50) P. vivax malaria and healthy controls (n = 50). Patient plasma concentrations of IL-4, IL-2, CXCL10, IL-1β, TNF-α, CCL2, IL-17A, IL-6, IL-10, IFN-γ, IL-12p70, CXCL8 and active TGF-β1 were determined through flow cytometry. The levels of several cytokines and chemokines, CXCL10, IL-10, IL-6, IL-4, CCL2 and IFN-γ were found to be significantly higher in severe, compared to non-severe P. vivax malaria patients. Severe thrombocytopenia was positively correlated with IL-4, CXCL10, IL-6, IL-10 and IFN-γ levels, renal dysfunction was related to an increase in IL-2, IL-1β, IL-17A and IL-8, and hepatic impairment with CXCL10, MCP-1, IL-6 and IFN-γ. A Lasso regression model suggests that IL-4, IL-10, CCL2 and TGF-β might be developed as biomarkers for severity in P. vivax malaria. Severe P. vivax malaria patients present specific cytokine and chemokine profiles that are different from non-severe patients and that could potentially be developed as biomarkers for disease severity.

Circulatory cytokeratin 17, marginal zone B1 protein and leucine-rich α2-glycoprotein-1 as biomarkers for disease severity and fibrosis in systemic sclerosis patients.

IntroductionSystemic sclerosis (Ssc) is a multiorgan debilitating autoimmune disease that associates the triad: vascular involvement, tissue fibrosis and profound immune response alterations. Numerous previous studies focused on identification of candidate proteomic Ssc biomarkers using mass-spectrometry techniques and a large number of candidate Ssc biomarkers emerged. These biomarkers must firstly be confirmed in independent patient groups. The aim of the present study was to investigate the association of cytokeratin 17 (CK17), marginal zone B1 protein (MZB1) and leucine-rich α2-glycoprotein-1 (LRG1) with clinical and biological Ssc characteristics.Material and methodsSerum CK17, MZB1 and LRG1 were assessed in samples of the available Ssc biobank comprising of samples from 53 Ssc patients and 26 matched age and gender controls.ResultsCirculatory CK17, LRG1 and MZB1 concentrations were increased in Ssc patients. Cytokeratin 17 is independently associated with Ssc disease activity. Patients with pulmonary fibrosis expressed higher LRG1 and MZB1 concentrations. Serum MZB1 concentrations were also associated with extensive skin fibrosis.ConclusionsSerum CK17, MZB1 and LRG1 were confirmed biomarkers for Ssc. LRG1 seems a good biomarker for pulmonary fibrosis, while MZB1 is a good biomarker for extensive skin fibrosis. CK17 proved to be independently associated with Ssc disease severity, higher CK17 values being protective for a more active disease.

Interleukin-17, a salivary biomarker for COVID-19 severity.

T-helper 17 cell-mediated response and their effector IL-17 cytokine induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is a major cause of COVID-19 disease severity and death. Therefore, the study aimed to determine if IL-17 level in saliva mirrors its circulatory level and hence can be used as a non-invasive biomarker for disease severity. Interleukin-17 (IL-17) level was evaluated by ELISA in saliva and blood of 201 adult COVID-19 patients with different levels of severity. The IL-17 saliva level was also associated with COVID-19 disease severity, and need for mechanical ventilation and/or death within 29 days after admission of severe COVID-19 patients. We found that IL-17 level in saliva of COVID-19 patients reflected its circulatory level. High IL-17 level in saliva was associated with COVID-19 severity (P<0.001), need for mechanical ventilation (P = 0.002), and/or death by 29 days (P = 0.002), after adjusting for patients' demographics, comorbidity, and COVID-19 serum severity markers such as D-Dimer, C-reactive protein, and ferritin. We propose that saliva IL-17 level could be used as a biomarker to identify patients at risk of developing severe COVID-19.

Pedometers and Accelerometers in Multiple Sclerosis: Current and New Applications.

Pedometers and accelerometers have become commonplace for the assessment of physical behaviors (e.g., physical activity and sedentary behavior) in multiple sclerosis (MS) research. Current common applications include the measurement of steps taken and the classification of physical activity intensity, as well as sedentary behavior, using cut-points methods. The existing knowledge and applications, coupled with technological advances, have spawned new opportunities for using those motion sensors in persons with MS, and these include the utilization of the data as biomarkers of disease severity and progression, perhaps in clinical practice. Herein, we discuss the current state of knowledge on the validity and applications of pedometers and accelerometers in MS, as well as new opportunities and strategies for the improved assessment of physical behaviors and disease progression, and consequently, personalized care.

Pediatric Moyamoya Biomarkers: Narrowing the Knowledge Gap

Moyamoya is a progressive cerebrovascular disorder that leads to stenosis of the arteries in the distal internal carotid, proximal middle cerebral and proximal anterior cerebral arteries of the circle of Willis. Typically a network of collaterals form to bypass the stenosis and maintain cerebral blood flow. As moyamoya progresses it affects the anterior circulation more commonly than posterior circulation, and cerebral blood flow becomes increasingly reliant on external carotid supply. Children with moyamoya are at increased risk for ischemic symptoms including stroke and transient ischemic attacks (TIA). In addition, cognitive decline may occur over time, even in the absence of clinical stroke. Standard of care for stroke prevention in children with symptomatic moyamoya is revascularization surgery. Treatment of children with asymptomatic moyamoya with revascularization surgery however remains more controversial. Therefore, biomarkers are needed to assist with not only diagnosis but also with determining ischemic risk and identifying best surgical candidates. In this review we will discuss the current knowledge as well as gaps in research in relation to pediatric moyamoya biomarkers including neurologic presentation, cognitive, neuroimaging, genetic and biologic biomarkers of disease severity and ischemic risk.

SNHG3 deficiency restrains spinal cord injury-induced inflammation through sponging miR-139-5p and provides a novel biomarker for disease severity.

MicroRNAs and long non-coding RNAs play pivotal roles in the progression and recovery of spinal cord injury (SCI), which is a serious traumatic disease in central nervous system. The purpose of this study was to investigate the expression and clinical value of SNHG3 in SCI patients and explore the regulatory effects of SNHG3 on SCI-induced inflammatory responses in vitro. The relationship between SNHG3 and miR-139-5p was confirmed using a dual-luciferase reporter assay. A SCI cell model was constructed in SH-SY5Y cells using hypoxia treatment. SNHG3 and miR-139-5p expression was analyzed using qRT-PCR. Effects of SNHG3 and miR-139-5p on cell model viability and inflammatory cytokines were evaluated by CCK-8 assay and ELISA kits, respectively. ROC curves based on serum SNHG3 and miR-139-5p were constructed to evaluate their diagnostic performance. In SCI patients, serum SNHG3 was upregulated, but miR-139-5p was downregulated (P<0.05), and a negative correlation between the two ncRNAs was found. Both SNHG3 and miR-139-5p showed relatively high discrimination abilities for the screening of SCI and complete SCI (CSCI) patients. SNHG3 was positively correlated with inflammatory cytokines, and miR-139-5p showed opposite results in SCI patients. By in-vitro analysis, SNHG3 knockdown enhanced cell viability but inhibited inflammation by increasing miR-139-5p. All the data found that serum upregulated SNHG3 and downregulated miR-139-5p served as biomarkers to diagnose SCI and indicate injury severity. The deficiency of SNHG3 alleviated neuronal injury by restraining inflammatory responses through targeting miR-139-5p. Thus, the SNHG3/miR-139-5p axis may provide novel biomarkers and therapeutic targets for SCI.

Elevated serum human epididymis protein 4 is associated with disease severity and worse survival in idiopathic pulmonary fibrosis: a cohort study.

BackgroundElevated expression of human epididymis protein 4 (HE4) was previously described in connective tissue disease-associated interstitial lung diseases (CTD-ILDs) and cystic fibrosis (CF), but the clinical significance of HE4 has remained unknown in idiopathic pulmonary fibrosis (IPF), which is a progressive fibrosing ILD with a heterogeneous course that is in urgent need of reliable biomarkers in its clinical practice.MethodsA total of 27 IPF patients with acute exacerbation status (AE-IPF), 32 IPF patients with stable status (S-IPF), and 29 sex-age matched healthy controls were retrospectively included. The levels of serum HE4 and Krebs von den Lungen-6 (KL-6) of the 3 cohorts were measured. In addition, the pulmonary expression of HE4 was evaluated in lung transplant specimens of IPF using immunohistochemistry and Western blot, and noncancerous lung tissue resected from early-stage lung cancer patients as controls. The endpoint of follow-up was March 1st, 2022, and the Cox regression model was used to analyze the prognostic value of HE4.ResultsThe levels of HE4 and KL-6 were obviously elevated in AE-IPF patients compared to S-IPF (296.4 vs. 178.1 pmol/L for HE4, P<0.001; 2,007.0 vs. 990.5 IU/mL for KL-6, P<0.001) or healthy controls (296.4 vs. 51.8 pmol/L for HE4, P<0.001; 2,007.0 vs. 181.0 IU/mL for KL-6, P<0.001). Significant correlations were observed between serum HE4 levels and percent predicted diffusing capacity of the lung for carbon monoxide (DLCO%) (r=−0.526, P<0.001), percent predicted forced vital capacity (FVC%) (r=−0.344, P=0.024), gender-age-physiology (GAP) index (r=0.535, P<0.001), and oxygenation index (r=−0.550, P<0.001) in IPF patients. In histological analysis, overexpression of HE4 in mucosal epithelium of dilated bronchi was observed in IPF patients compared with controls. Multivariate cox regression revealed that serum levels of HE4 [hazard ratio (HR) =1.004, P=0.042] and GAP index (HR =1.374, P=0.010) were associated with worse survival in IPF patients.ConclusionsThe expression of serum HE4 was obviously elevated in IPF patients, especially in AE-IPF patients. In addition, serum HE4 could be utilized as a biomarker of disease severity and poor prognosis of IPF patients. These findings warrant further validation in larger, multi-center, and longitudinal cohorts.

Preoperative Lymphocyte to Monocyte Ratio as a Predictive Biomarker for Disease Severity and Spinal Fusion Failure in Lumbar Degenerative Diseases Patients Undergoing Lumbar Fusion

ObjectiveThis study was designed to determine whether lymphocyte to monocyte ratio (LMR) correlated with the intervertebral disc degeneration (IDD) severity and the postoperative spinal fusion rate in patients with lumbar disc disease.Methods303 patients undergoing posterior lumbar decompression and fusion were retrospectively analyzed. An examination of the blood count was performed before surgery. The cumulative grade was calculated by summing the pfirrmann grades of all lumbar discs. Grouping was based on the 50th percentile of cumulative grade and spinal fusion. The relationship between LMR and IDD severity and spinal fusion was explored using correlation analyses and logistic regression models. The receiver operating characteristic (ROC) curve was performed to measure model discrimination, and Hosmer-Lemeshow (H-L) test was used to measure calibration. Meanwhile, the ROC curve evaluated the discrimination ability of LMR in predicting severe degeneration and fusion failure.ResultsLMR was significantly lower in the severe degeneration group (cumulative grade > 18) than in the mild to moderate degeneration group (cumulative grade ≤ 18). Furthermore, the LMR of the fusion group was significantly higher than that of the non-fusion group. The multivariate binary logistic models revealed that LMR was an independently influencing factor of the severe degeneration and fusion failure (OR: 0.793, 95% CI: 0.638–0.987, p = 0.038; OR: 0.371, 95% CI: 0.258–0.532, p < 0.001). The models showed excellent discrimination and calibration. The area under the curve (AUC) of severe degeneration and fusion failure identified by LMR were 0.635 and 0.643, respectively, and the corresponding cut-off values were 3.16 and 3.90.ConclusionLMR is significantly associated with the risk of severe disc degeneration and spinal fusion failure.

Determination of reference intervals for neutrophil granular enzymes is affected by cell isolation techniques

ObjectivesNeutrophils and their granular enzymes such as neutrophil elastase (NE) and myeloperoxidase (MPO) play important roles in inflammatory diseases, and might be utilized as biomarkers for disease severity and progression. The aim of this study was to determine reference intervals for NE and MPO activity in healthy volunteers comparing two methods of neutrophil isolation. MethodsNeutrophils were isolated using ficoll density gradient centrifugation or immunomagnetic negative selection in two separate volunteers' cohorts. Subsequently, cells were lysed and incubated with chromogens for NE and MPO activity measurements, then measured with a microplate reader at 415 or 450 nm respectively. ResultsThe enzymatic activity of NE and MPO depended on the neutrophil isolation technique. Both enzymatic activities were significantly higher (P < 0.001) after isolating neutrophils with ficoll density gradient centrifugation than using the immunomagnetic negative selection. ConclusionsWe demonstrated that neutrophil isolation is an important factor that influences the outcome of enzymatic activity measurements. Techniques based on immunomagnetic negative selection are favorable, specifically for investigations related to NE and MPO activity. When using NE and MPO activity measurements in clinical practice, care must be taken to interpret the data depending on the applied cell isolation technique.

GDF-15 and sST-2 act as biomarkers of disease severity but not independent predictors in idiopathic membranous nephropathy

BackgroundWe aimed to explore biomarkers of disease severity in idiopathic membranous nephropathy (IMN) and independent predictors of prognosis in IMN. MethodsClinical data were collected from 79 IMN patients. Serum levels of growth differentiation factor-15 (GDF-15) and soluble suppression of tumorigenicity (sST-2) were tested by enzyme-linked immunosorbent assay (ELISA) in IMN patients and subgroups, and correlation analysis was performed. Univariate and multiple logistic regression analyses were performed to identify independent predictors of IMN, and a combined-factors model was constructed. Moreover, the area under the receiver operating characteristic (ROC) curve (AUC) was used to evaluate the prognostic efficacy. ResultsThe levels of GDF-15 were significantly higher in the IMN group and subgroups with low estimated glomerular filtration rate (eGFR) and high 24 hour-urine protein (24 h-UP), whiles sST-2 level was only significantly higher in the IMN group. GDF-15 levels were positively correlated with creatinine (Crea), cystatin C (Cys-C) and 24 h-UP and negatively correlated with GFR and albumin (Alb), while sST-2 levels were positively correlated with Urea and Cys-C and negatively correlated with eGFR. After one year of follow-up, 54 patients had incomplete remission. Serum phospholipase A2 receptor antibody (PLA2R-Ab), Urea, high-density lipoprotein cholesterol (HDL-C) and 24 h-UP but not GDF-15 and sST2 were independent predictors of prognosis in IMN patients, but combined factors showed the best prognostic efficacy. ConclusionSerum levels of GDF-15 and sST-2 may be potential biomarkers for the severity of IMN, while the combined-factors model is effective for predicting the risk factors of incomplete remission in IMN.