Integrated Bioinformatics Exploration and Preliminary Clinical Verification for the Identification of Crucial Biomarkers in Severe Cases of COVID-19.

Background: Coronavirus disease 2019 (COVID-19) is a respiratory infectious illness caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The objective of this study is to identify reliable and accurate biomarkers for the early stratification of disease severity, a crucial aspect that is currently lacking for the impending phases of the next COVID-19 pandemic. Methods: In this study, we identified important module and hub genes related to clinical severe COVID-19 using differentially expressed genes (DEGs) screening combing weighted gene co-expression network analysis (WGCNA) in dataset GSE213313. We further screened and confirmed these hub genes in another two new independent datasets (GSE172114 and GSE157103). In order to evaluate these key genes' stability and robustness for diagnosing or predicting the progression of illness, we used RT-PCR validation of selected genes in blood samples obtained from hospitalized COVID-19 patients. Results: A total of 968 and 52 DEGs were identified between COVID-19 patients and normal people, critical and non-critical patients, respectively. Then, using WGCNA, 10 modules were constructed. Among them, the blue module positively associated with clinic disease severity of COVID-19. From overlapped section between DEGs and blue module, 12 intersected common differential genes were obtained. Subsequently, these hub genes were validated in another two new independent datasets as well and 9 genes that overlapped showed a highly correlation with disease severity. Finally, the mRNA expression levels of these hub genes were tested in blood samples from COVID-19 patients. In severe cases, there was increased expression of MCEMP1, ANXA3, CD177, and SCN9A. In particular, MCEMP1 increased with disease severity, which suggested an unfavorable development and a frustrating prognosis. Conclusion: Using comprehensive bioinformatical analysis and the validation of clinical samples, we identified four major candidate genes, MCEMP1, ANXA3, CD177, and SCN9A, which are essential for diagnosis or development of COVID-19.

TREM-1, TREM-2 and their association with disease severity in patients with COVID-19.

BACKGROUND: Delayed diagnosis and inadequate treatment caused by limited biomarkers are associated with the outcomes of COVID-19 patients. It is necessary to identify other promising biomarkers and candidate targets for defining dysregulated inflammatory states. METHODS: The triggering receptors expressed on myeloid cell (TREM)-1 and TREM-2 expression from hospitalized COVID-19 patients were characterized using ELISA and flow cytometry, respectively. Their correlation with disease severity and contrast with the main clinical indicators were evaluated. RESULTS: Increased expression of soluble TREM-1 and TREM-2 in the plasma of COVID-19 patients was found compared to the control group. Moreover, membrane-bound TREM-1 and TREM-2 expression was upregulated on the cell surface of circulating blood T cells from COVID-19 patients. Correlation analysis showed that sTREM-2 levels were negatively correlated with PaO2/FiO2, but positively correlated with C-reactive protein (CRP), procalcitonin (PCT) and interleukin (IL)-6 levels. Receiver operating characteristic curve analysis indicated that the predictive efficacy of sTREM-1 and sTREM-2 was equivalent to CRP and IL-6, and a little better than absolute leukocyte or neutrophil count and PCT in distinguishing disease severity. CONCLUSION: TREM-2 and TREM-1 are critical host immune factors that response to SARS-COV-2 infection and could serve as potential diagnostic biomarkers and therapeutic targets for COVID-19.

The expression of soluble TREM-1 and TREM-2 in plasma and membrane-bound TREM-1 and TREM-2 on the cell surface was upregulated in COVID-19 patients.sTREM-2 level was negatively correlated with PaO₂/FiO₂, but positively correlated with CRP, PCT and IL-6 level, respectively.sTREM-1 and sTREM-2 exhibited potential predictive abilities, and their expression was equivalent to CRP and IL-6, and better than the absolute leukocytes or neutrophil counts and PCT in distinguishing disease severity.

Clinical Effect of Bushen Huoxue Method Combined with Platelet-Rich Plasma in the Treatment of Knee Osteoarthritis and Its Effect on IL-1, IL-6, VEGF, and PGE-2.

Objective: To observe the clinical efficacy of the Bushen Huoxue method combined with platelet-rich plasma (PRP) in the treatment of knee osteoarthritis (KOA) and its effect on serum and joint fluid interleukin-1 (IL-1), interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), and prostaglandin E2 (PGE-2). Methods: A total of 64 cases of KOA diagnosed and treated from January 2020 to January 2021 were randomly divided into research group and control group, with 32 cases in each group. The control group was treated with PRP, and the research group took the prescription of the Bushen Huoxue method on the basis of the control group. The clinical efficacy was assessed according to the criteria in "the diagnosis and Treatment of osteoarthritis," osteoarthritis index score and pain visual analogue score (VAS). Serum and articular fluid VAS, IL-1, IL-6, VEGF, and PGE-2 levels were detected by the enzyme-linked immunosorbent assay (ELISA). Results: The clinical effective rate of the research group was 93.8%, which was significantly higher than that of the control group (90.6%). There was no significant difference in the scores of osteoarthritis index between the two groups before treatment, but the scores of both groups decreased after treatment and was lower in the research group than those in the control group. VAS was significantly decreased in two groups after treatment and it was lower in the research group than that in the control group. After treatment, the levels of IL-1, IL-6, and PGE-2 in serum and articular fluid all indexes were decreased, and the levels in the research group were lower than those in the control group. Conclusions: PRP joint cavity injection combined with oral administration of Bushen Huoxue prescription, and PRP joint cavity injection alone can improve the efficacy of KOA, relieve knee pain, and promote the recovery of knee function. The mechanism may be related to the reduction of IL-1, IL-6, VEGF levels, and PGE-2 levels in the serum and joint fluid. However, the efficacy of combination therapy was superior to PRP alone.

Efficacy of Buqi Huoxue Decoction Combined with Cardiac Rehabilitation Nursing after Coronary Intervention in Patients with Acute ST-Segment Elevation Myocardial Infarction and Its Influence on Prognosis.

Objective: To investigate the efficacy of the application of Buqi Huoxue Decoction combined with cardiac rehabilitation nursing for patients with acute ST-segment elevation myocardial infarction (STEMI) after percutaneous coronary intervention (PCI) and its influence on the prognosis. Methods: 120 STEMI patients undergoing PCI were randomly divided into control group, cardiac care group, traditional Chinese medicine and western medicine group (TCM + WM group), and comprehensive treatment group. The control group was treated with a conventional antiplatelet therapy. On the basis of the control group, the cardiac care group was combined with cardiac care treatment. The TCM + WM group was combined with Buqi Huoxue Decoction, and the comprehensive treatment group was combined with cardiac rehabilitation care and Buqi Huoxue Decoction. The total clinical effective rate, readmission rate, and adverse reaction rate of the four groups were measured. Moreover, the myocardial injury markers (creatine kinase isoenzyme (CK-MB), cardiac troponin I (cTnI), and α-Hydroxybutyrate dehydrogenase (α-HBDH)), vascular endothelial function indexes (endothelin (ET-1) and vascular endothelial growth factor (VEGF)), cardiac function indexes (left ventricular ejection fraction (LVEF), left ventricle shortening rate (LFS), left ventricular end diastolic diameter (LVEDd), and left ventricular end systolic diameter (LVESd)), and QOL quality of life score (appetite, spirit, sleep, fatigue, and daily life) were measured. Results: The total effective rate of comprehensive treatment group was obviously increased versus to the control group and cardiac care group. The CK-MB, cTnI, α-HBDH, ET-1, LVEDd, and LVESd levels and SAS and SDS scores in the four groups were decreased, and VEGF, LVEF, and FS levels and QOL quality of life scores were increased after treatment. Moreover, the comprehensive treatment group has more significant changes than the other three groups. The readmission rate in comprehensive treatment group was significantly lower than the other three groups, and the difference in the incidence of adverse reactions in the four groups was not statistically significant. Conclusion: Buqi Huoxue Decoction combined with cardiac rehabilitation after PCI has a significant clinical effect on STEMI patients with PCI postoperative treatment, which can effectively reduce myocardial injury, improve the patient's cardiac function and vascular endothelial function, and improve the patient's quality of life, which can better improve the prognosis of patients.

Low-dose cadmium activates the JNK signaling pathway in human renal podocytes.

Cadmium (Cd) is an environmental toxin. Our previous study demonstrated that low­dose Cd damages the integrity of the glomerular filtration barrier (GFB); however, the underlying mechanisms are poorly understood. Podocytes are a major component of the GFB, which regulate the passage of proteins. The present study aimed to investigate the effects of low­dose Cd on human renal podocytes (HRPs). HRPs were treated with Cd and activation of the c-Jun N-terminal kinase (JNK) pathway was examined by western blot analysis. Proliferation, viability and apoptosis of HRPs were evaluated by MTT assay, trypan blue exclusion assay and flow cytometry, respectively. The properties of HRPs were validated by immunofluorescence staining and Phalloidin­labeling. The results indicated that 4 µM Cd may activate the JNK pathway, and increase the protein expression levels of c­Jun and c­Fos. However, proliferation, viability, apoptosis and alignment of the F­actin cytoskeleton in HRPs were not significantly affected by Cd treatment, with or without SP600125 pretreatment. In addition, the expression levels of CD2­associated protein and synaptopodin, which are differentiation markers of HRPs, remained unchanged following Cd treatment. These results indicated that low­dose Cd activates the JNK pathway but does not significantly affect HRP function.

Dihydroartemisinin ameliorates sepsis-induced hyperpermeability of glomerular endothelium via up-regulation of occludin expression.

Sepsis, the systemic inflammatory responses after infection, remains a serious cause of morbidity and mortality in critically ill patients. The anti-malarial agent dihydroartemisinin (DHA) has been shown to be anti-inflammatory. In this study, we examined the effects of DHA on sepsis-induced acute kidney injury (AKI) and explored the mechanism underlying its mode of action in AKI. In a lipopolysaccharide (LPS)-induced mouse model, we observed that DHA treatment ameliorated glomerular injury, and relieved elevation of the urine albumin to creatinine ratio (UACR) and serum creatinine. At a concentration of 25 µM, DHA had no effect on overall cellular viability or apoptosis in assays with human renal glomerular endothelial cells (HRGECs), but significantly inhibited the tumor necrosis factor-α (TNF-α)-induced hyperpermeability of HRGEC monolayers. We found that TNF-α decreases the expression of the junctional protein occludin in HRGECs, which is reversed by DHA. Taken together, our results demonstrate that DHA decreases permeability of the glomerular endothelium by maintenance of occludin expression. This suggests DHA may have therapeutic utility in sepsis-induced AKI.

CD109 expression is upregulated in penile squamous cell carcinoma.

Cluster of differentiation (CD) 109 is a glycosylphosphatidylinositol-anchored cell surface glycoprotein and a co-receptor for transforming growth factor ß. The expression of CD109 has been detected in squamous cell carcinoma of the lung, esophagus, skin and gallbladder. The aim of the present study was to evaluate CD109 expression in penile squamous cell carcinoma (PSCC). CD109 expression in PSCC tumor and adjacent tissues from 45 specimens in tissue microarrays was examined by immunohistochemical analyses. In addition, 3 fresh surgical samples of PSCC were collected and examined for their CD109 mRNA and protein levels by reverse transcription-quantitative polymerase chain reaction, western blotting and immunofluorescence staining. CD109 transcription and expression were significantly higher in the PSCC tissues compared with adjacent normal penile tissues, and its expression was restricted to squamous cells. However, CD109 expression level was not associated with the PSCC differentiation grade. These results suggest that CD109 may be associated with the pathogenesis of PSCC, and may therefore be a potential therapeutic target.

Erratum: Dihydroartemisinin induces endothelial cell anoikis through the activation of the JNK signaling pathway.

[This corrects the article DOI: 10.3892/ol.2016.4870.].

The role of Notch signalling in ovarian angiogenesis.

In adults, the ovary is characterized with extensive angiogenesis and regular intervals of rapid growth. Ovarian function is dependent on the network of angiogenic vessels which enable the follicle and/or corpus luteum to receive oxygen, nutrients and hormonal support. Abnormal angiogenesis is involved in the induction and development of pathological ovary, such as polycystic ovary syndrome and ovarian cancer. Notch signalling pathway is one of the primary regulators of angiogenesis and a therapeutic target for ovarian diseases. Here, we will review literatures on the expression pattern of Notch pathway components in the ovary and on the role of Notch signalling pathway on ovarian angiogenesis.

GATA3-induced vWF upregulation in the lung adenocarcinoma vasculature.

Lung adenocarcinoma (LAC) is the leading cause of cancer-related death worldwide. Aberrant expression of genes expressed preferentially in the lung tumor vasculature may yield clues for prognosis and treatment. Von Willebrand factor (vWF) is a large multifunctional glycoprotein with a well-known function in hemostasis. However, vWF has been reported to exert an anti-tumor effect, independent of its role in hemostasis. We investigated the expression of vWF in LAC through immunohistochemical staining of tumor tissue microarrays (TMAs). We found that vWF was overexpressed preferentially in the tumor vasculature of LAC compared with the adjacent tissue vasculature. Consistently, elevated vWF expression was found in endothelial cells (ECs) of fresh human LAC tissues and transplanted mouse LAC tissues. To understand the mechanism underlying vWF up-regulation in LAC vessels, we established a co-culture system. In this system, conditioned media (CM) collected from A549 cells increased vWF expression in human umbilical vein endothelial cells (HUVECs), suggesting enhanced expression is regulated by the LAC secretome. Subsequent studies revealed that the transcription factor GATA3, but not ERG, a known regulator of vWF transcription in vascular cells, mediated the vWF elevation. Chromatin immunoprecipitation (ChIP) assays validated that GATA3 binds directly to the +220 GATA binding motif on the human vWF promoter and A549 conditioned media significantly increases the binding of GATA3. Taken together, we demonstrate that vWF expression in ECs of LAC is elevated by the cancer cell-derived secretome through enhanced GATA3-mediated transcription.

Low Dose Cadmium Inhibits Proliferation of Human Renal Mesangial Cells via Activation of the JNK Pathway.

Cadmium (Cd) is a heavy metal and environmental pollutant. The kidney is the principal target organ of Cd exposure. Previously, we found that low concentration of Cd damages the integrity of the glomerular filtration barrier. However, little is known about the effects of Cd on renal mesangial cells, which provide structural support for the glomerular capillary loops and regulate intraglomerular blood flow. In this study, human renal mesangial cells (HRMCs) were cultured in the presence of serum and treated with 4 µM Cd. We found that Cd activates the c-Jun N-terminal kinase (JNK) pathway, and increases the protein levels of c-Jun and c-Fos. Cd treatment also induces a decrease in proliferation and an increase in apoptosis of HRMCs, but only the decrease in HRMC proliferation was reversed by pretreatment with SP600125, an inhibitor of the JNK pathway. In addition, Cd does not change the expression of α-smooth muscle actin and platelet-derived growth factor receptor-ß, the markers of mesangial cells, or the alignment of the filamentous actin (F-actin) cytoskeleton of HRMCs. Our data indicate that the JNK pathway mediates the inhibitory effects of Cd on HRMC proliferation.

Dihydroartemisinin induces endothelial cell anoikis through the activation of the JNK signaling pathway.

Angiogenesis is required for the growth and metastasis of solid tumors. The anti-malarial agent dihydroartemisinin (DHA) demonstrates potent anti-angiogenic activity, but the underlying molecular mechanisms are not yet fully understood. During the process of angiogenesis, endothelial cells migrating from existing capillaries may undergo programmed cell death after detaching from the extracellular matrix, a process that is defined as anchorage-dependent apoptosis or anoikis. In the present study, DHA-induced cell death was compared in human umbilical vein endothelial cells (HUVECs) cultured in suspension and attached to culture plates. In suspended HUVECs, the cell viability was decreased and apoptosis was increased with the treatment of 50 µM DHA for 5 h, while the same treatment did not affect the attached HUVECs. In addition, 50 µM DHA increased the phosphorylation of c-Jun N-terminal kinase (JNK) in suspended HUVECs, but not in attached HUVECs, for up to 5 h of treatment. The JNK inhibitor, SP600125, reversed DHA-induced cell death in suspended HUVECs, suggesting that the JNK pathway may mediate DHA-induced endothelial cell anoikis. The data from the present study indicates a novel mechanism for understanding the anti-angiogenic effects of DHA, which may be used as a component for chemotherapy.