Evaluation of Serum Levels of MicroRNA-200C and ACE2 Gene Expression in Severe and Mild Phases of Patients with COVID-19.

The role of microRNA (miR)200c-3p in regulating ACE2 gene expression in viral and bacterial respiratory diseases has been established. Since ACE2 reduces the acute inflammatory effects in lung diseases and acts as a coronavirus receptor to invade the lung cells, this study investigates the relationship between miR-200c-3p and ACE2 expression in COVID -19 patients. In this study, COVID-19 patients were divided into two groups: mild phase (PCR-positive and mild symptoms) and severe phase (PCR-positive with acute pulmonary symptoms and inflammation). Then, the subjects' demographic, clinical, and paraclinical characteristics were recorded using a prepared checklist. Total RNA was isolated from all samples according to the Trizol kit protocol to evaluate gene expression. Subsequently, the extracted product was analysed for miR-200c expression and ACE2 target gene expression by real-time PCR. The results of the checklist data showed that smoking, cough, and the factors ESR and HCT were statistically significant between the two groups of patients in the mild and acute phases. Also, the mean expression of the miR-200c gene in the mild and acute patients was 1.87±0.70 and 1.87±0.62, respectively, which was not statistically significant. Still, the mean expression of the ACE2 gene, which was 3.96±0.76 and 3.28±0.52 in the mild and acute disease groups, respectively, showed a significant difference between the two groups. This study showed that the expression levels of ACE2 were significantly reduced in people with severe inflammation compared to people with mild inflammation.

Serum levels of bone sialoprotein, osteopontin, and ß2-microglobulin in stage I of multiple myeloma.

CONTEXT: The fluctuations of proteins in multiple myeloma (MM) are well-known markers for checking the status of the patients. AIMS: The objective of this study was to examine three proteins that have an important role in disease progression. SUBJECTS AND METHODS: The study was performed with two groups: 30 MM stage I patients' (14 females/16 males; aged 60.83 ± 12.38 years) as case group and 40 healthy individuals (18 females/22 males; aged 57.65 ± 6.43 years) as control group. Both groups have been matched in gender and age. Bone sialoprotein (BSP), osteopontin (OPN), and ß2-microglobulin (ß2M) were measured with an enzyme-linked immunosorbent assay. RESULTS: Serum BSP levels of MM-I patients was significantly higher than that of healthy controls (29.24 ± 5.57 vs. 20.89 ± 3.67, P = 0.001). OPN levels of MM-I patients were significantly lower than that of healthy individuals (12.03 ± 3.45 vs. 19.35 ± 4.67, P = 0.001). ß2M levels of patients and controls were similar (1.49 ± 0.67 vs. 1.29 ± 0.55, P = 0.193). CONCLUSIONS: The results suggested that myeloma cells may affect the production of BSP and OPN, which possibly contributes to osteoclastic bone resorption in MM-I patients. Their levels may be a useful biomarker for assessing bone destruction in MM-I patients and distinguishing MM-I from healthy individuals.

The Effects of Nano-curcumin Supplementation on Serum Level of hs-CRP, Adhesion Molecules, and Lipid Profiles in Hemodialysis Patients, A Randomized Controlled Clinical Trial.

INTRODUCTION: Hemodialysis (HD) patients are considered as a high-risk population for cardiovascular disease, within which morbidity and mortality have been determined to be associated with dyslipidemia, pro-inflammatory cytokines, increased levels of C-reactive protein (CRP), and adhesion molecules (ICAM-1, VCAM-1). Different markers have been investigated to detect inflammation in hemodialysis patients, as well as the prognostic values of these markers. METHODS: The present study aimed to investigate the effect of nano-curcumin (120 mg) over 12 weeks on hs-CRP levels, adhesion molecules (ICAM-1, VCAM-1), and serum lipid profiles on hemodialysis patients in a randomized controlled clinical trial. RESULTS: The results revealed that the mean serum hs-CRP level in the nano-curcumin group exhibited a decrease by the end of the study, when compared to mean serum hs-CRP level in the placebo group. However, this between-group trend was not found to be statistically significant (P > .05). Nevertheless, a significant difference was determined between the values in the group receiving nano-curcumin, in comparison with the placebo group, at the end of the study (P < .001). Based on the attained results, mean serum levels of VCAM-1 in the nano-curcumin group were significantly reduced at the end of the study, compared with the placebo group (P < .001). Furthermore, the between-group changes comparison showed significant reductions in serum levels of ICAM- 1 in patients treated with nano-curcumin at the end of the study (P < .05). Additionally, though decreases in mean triglycerides, total cholesterol, LDL-C were noted, there were no statistically significant between-group differences (P > .05). Moreover, between-group changes comparison of HDL-C levels and fasting blood sugar did not show any significant changes. CONCLUSION: The current study indicates that nano-curcumin may show beneficial effects in lowering inflammation and hs-CRP levels, as well as adhesion molecules (ICAM-1, VCAM-1), in hemodialysis patients. However, the evidence is still insufficient.

Titanium dioxide nanoparticles induce endothelial cell apoptosis via cell membrane oxidative damage and p38, PI3K/Akt, NF-κB signaling pathways modulation.

BACKGROUND: Titanium dioxide nanoparticles (TiO2 NPs) are widely used nanoparticles. Despite, several studies investigated the toxic effects of TiO2 NPs on HUVECs, the results are contradictory and the possible underlying mechanisms remain unclear. METHODS: In the present study, we conducted an in vitro study to re-evaluate the possible toxic effects of TiO2 NPs on HUVECs including cell viability, lipids peroxidation, intracellular signaling pathways and nitric oxide syntheses enzymes. RESULTS: Our results demonstrated that, TiO2 NPs were internalized to HUVECs and induce intracellular reactive oxygen species production and cell membrane oxidative damage at the higher concentration. TiO2 NPs induce IKKα/ß and Akt phosphorylation and p38 dephosphorylation. After 24 h treatment, pro-inflammatory cytokines, adhesion molecules and chemokine upregulated significantly. TiO2 NPs have no significant effects on eNOS enzymatic activation and iNOS gene expression. At cellular level, apoptosis is the main process that occur in response to TiO2 NPs treatment. HUVECs pretreatment with N-acetyl-l-cysteine (NAC) ameliorate the toxic effects of TiO2 NPs that indicate the oxidative stress is essential in TiO2 NPs -induced toxicity. Total antioxidant capacity show a trend to increase in response to TiO2 NPs exposure. CONCLUSIONS: Taken together, this study confirmed the effects of TiO2 NPs on endothelial cells and proposed multiple underlying mechanisms including cell membrane oxidative damage and intracellular processes.

Increased levels of angiogenic factors in microvascular angina.

BACKGROUND: Recent studies have suggested that angiogenic factors may affect vascular endothelial integrity. On the other hand, endothelial dysfunction is the main pathological mechanism in microvascular angina (MVA) or cardiac syndrome X. Therefore, we aimed to determine the levels of angiogenic factors in MVA patients. In addition, we investigated the effects of metoprolol, as a beta blocker agent, on the serum levels of these factors. METHODS: Thirty patients with MVA (17 female/13 male; mean age: 55.53±9.18 years) and twenty healthy controls (14 female/6 male; mean age: 51.40±9.16 years) were enrolled.The serum amounts of angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2) and tyrosine kinase-2 receptor (Tie-2) were measured in healthy controls, MVA patients at baseline and after metoprolol therapy (25 mg for one month) by enzyme-linked immunosorbent assay. RESULTS: The levels of Ang-2 and Tie-2 were significantly higher in MVA patients at baseline in comparison with controls (Ang-2: 277.02±186.08 vs.164.46±49.83 ng/l, P=0.011; Tie-2: 28.97±18.85 vs. 14.90±4.05 ng/ml, P=0.002; respectively). But this difference in the Ang-1 levels was not significant (P=0.829). Additionally, the levels of angiogenic factors in MVA patients after metoprolol therapy were not significantly changed in comparison with the baseline status (P>0.05). CONCLUSION: Our results considered a possible role for angiogenic factors in the pathophysiology of MVA, which need further investigation for elucidation. In addition, this study has not showed an effective role for metoprolol in changing the angiogenic factors levels as a therapeutic agent in MVA.

Complete inhibition of phosphatase and tensin homolog promotes the normal and oxygen-glucose deprivation/reperfusion-injured PC12 cells to cell death.

Introduction: Lipid phosphatase and tensin homolog deleted from chromosome 10 (PTEN) antagonizes phosphoinositide 3-kinase (PI3K)/AKT cell survival pathway. The effect of PTEN inhibitors has been rarely examined on cell survival following reperfusion injury. In this study, we investigated the neuroprotective effect of SF1670, as a new PTEN inhibitor, on an in vitro stroke-like model. Methods: PC12 cells were exposed to oxygen-glucose deprivation/reperfusion (OGD/R). The cells were treated in five conditions as follows: normoxic normoglycemic (NO/NG); 60 minutes OGD; 60 minutes OGD and 6 h reperfusion (OGD/R); OGD/R treated with 10 µM SF1670 (OGD/R-SF), and NO/NG treated with 10 µM SF1670 (NO/NG-SF). Then, phosphorylation levels of AKT, P38 in PC12 cells were measured by immunoblotting. The cell viability was also determined by colorimetric assay. Results: The results of immunoblotting revealed that following OGD/R the levels of phospho-AKT (p-AKT) significantly decreased, compared to NO/NG cells (P < 0.05). However, the ratio of p-AKT/total AKT significantly increased in the presence of SF1670 in the OGD/R-SF group, compared to the OGD/R condition. On the other hand, SF1670 significantly reduced the p-P38 MAPK and p-JNK levels, compared to OGD/R cells. Moreover, cell viability significantly decreased in the OGD and OGD/R condition compared to NO/NG cells. Surprisingly, SF-treated cells (OGD/R-SF and NO/NG-SF group) showed low cell viability compared to NO/NG condition. Conclusion: Overall, our results demonstrated that complete inhibition of phosphatase activity of PTEN not only did not exhibit neuroprotective effect but also promoted PC12-deprived cells to death.

Transcriptional activity of tumor necrosis factor-alpha gene in peripheral blood mononuclear cells in patients with coronary slow flow.

BACKGROUND: Coronary slow flow (CSF), an angiographic phenomenon that is characterized by a delayed coronary blood flow in the absence of obstructive coronary artery stenosis, is known as a disorder of the coronary microcirculation. Inflammation has an important role in the vascular hemostasis and endothelial dysfunction especially regarding monocyte adhesion and infiltration. Pro-inflammatory cytokines released by inflammatory cells result in endothelial cell dysfunction and cardiovascular diseases. It has been demonstrated that tumor necrosis factor-alpha (TNF-α) mainly influences the vascular homeostasis and endothelial dysfunction. In the present enquiry the transcriptional activity of TNF-α gene in peripheral blood mononuclear cells (PBMCs) of patients with CSF was compared with healthy controls in order to further survey the role of TNF-α in pathophysiology of CSF. METHODS: The study was carried out on 30 patients with CSF and 30 matched healthy controls. To analysis gene expression of TNF-α, total mRNA was isolated from PBMCs. The quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was used to compare the transcriptional activity of TNF-α gene between patients with CSF and controls. RESULTS: The mean ± standard error of mean of fold in CSF patients and controls were 0.20 ± 0.04 and 1.38 ± 0.27, respectively. The mRNA mean expressions of TNF-α (fold) were different in tested groups, which indicated a significant decrease in TNF-α in patients with CSF group (P = 0.0001). CONCLUSION: Expression of TNF-α was decreased in patients with CSF. Changes in TNF-α expression suggest a potential role for altered immune function in the pathophysiology of CSF.

Transcriptional Activity of Gene Encoding Subunits R1 and R2 of Interferon Gamma Receptor in Peripheral Blood Mononuclear Cells in Patients with Slow Coronary Flow.

BACKGROUND: Slow coronary flow (SCF) is a coronary artery disorder characterized with delayed opacification of epicardial coronary arteries without obstructive coronary disease. The pathophysiological mechanisms of SCF remain unclear. One of the possible mechanisms that may participate in the pathology of SCF is endothelial dysfunction related to the inflammatory process. Interferon gamma (IFN-γ) is an inflammatory cytokine that acts through its specific receptor composed of two subunits, IFN-γR1 and IFN-γR2. Transcriptional activity of the gene encoding these subunits influences IFN-γ activity. This study aimed to investigate the gene expression of IFN-γ receptor subunits in peripheral blood mononuclear cells (PBMC) from patients with SCF. METHODS: The study was performed with 30 patients (22 male/8 female) aged 35-76 (52.8±11.7 years) with SCF and 15 sex- (11 male/4 female), Body Max Index (BMI)- and age-matched (54.73±9.42 years) healthy subjects. Total mRNA was extracted from PBMC and was determined by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The relative expression values (2-ΔΔCt) between control and case groups were determined and the Mann-Whitney U test was used for statistical analysis. RESULTS: There was a significant increase in the gene expression of IFN-γR1 in PBMC from SCF patients vs. controls (P< 0.0001); but the differences in IFN-γR2 gene expression were statistically insignificant between patient and control groups (P= 0.853). CONCLUSIONS: It can be concluded that IFN-γ gene expression may influence the function of microvasculature and thereby contribute to the pathophysiology of SCF.

Protective effect of vitamin E against diabetes-induced oxidized LDL and aorta cell wall proliferation in rat.

UNLABELLED: Hyperlipidemia and oxidized-low-density lipoproteins (Ox-LDL) are important independent cardiovascular risk factors that have been shown to stimulate vascular smooth muscle cell (VSMC) proliferation. The purpose of the present study was to investigate the effect of vitamin E on Ox-LDL, lipid profile, C-reactive protein (CRP), and VSMC proliferation of rat aorta. METHODS: Male Wistar rats (n = 32) were divided into four groups namely: sham (SH), control (C), non-treated diabetic, and vitamin E-treated diabetic (VETD) groups. Ox-LDL, lipid profile, CRP and VSMC proliferation of aorta were measured after 42 days. RESULTS: The results revealed that along with a significant increase in VSMC proliferation, the amount of CRP, Ox-LDL, and lipid profiles in diabetic rats. VSMC proliferation was significantly ameliorated, and elevated CRP, Ox-LDL, and lipid profiles were also restored to those of shams in VETD. CONCLUSIONS: These findings strongly support the idea that diabetes induces Ox-LDL-mediated oxidative stress and VSMC proliferation in aorta of rat and imply that vitamin E has a strong protective effect as an antioxidant.

The Protective Effect of Vitamin E on Morphological and Biochemical Alteration Induced by Pre and Postnatal Ethanol Administration in the Testis of Male Rat Offspring: A Three Months Follow-up Study.

BACKGROUND: Dysmorphology and dysfunction caused by prenatal ethanol consumption in different organs of the offspring are wellknown phenomena. The objective of the present study was to explore the antioxidant effect of vitamin E supplementation on testis damage induced by maternal ethanol consumption during pregnancy and early postnatal days. METHODS: Pregnant Wistar rats on gestation day 7 were assigned to 3 groups, namely, control, ethanol and ethanol-vitamin E groups. Ethanol-treated rats received 4.5 g/kg BW ethanol once per day from day 7 and the procedure continued through postnatal day 21. Vitamin E group received 300 mg of vitamin E and the same amount of ethanol. The male offspring from each group were anesthetized by 10% chloral hydrate (0.5 ml/kg body weight) on day 21 and 90 (n=8 offspring form each group on day 21 and day 90). The results were analyzed by one-way ANOVA. A p<0.05 was considered significant. RESULTS: The results revealed significant (p<0.05) changes in oxidative stress parameters, luteinizing hormone and follicle-stimulating hormone, as well as testis structural alteration in offspring of ethanol group after 21 and 90 days of birth as compared to the control. Significant amelioration of changes in testis structure, along with restoration of the elevated level of oxidative stress parameters were found in vitamin E-treated animals. CONCLUSION: The findings revealed that prenatal and postnatal ethanol-induced toxicity in testis was exerted through oxidative stress and implied that these effects could be alleviated by vitamin E as an antioxidant.

Ethanol promotes rat aortic vascular smooth muscle cell proliferation via increase of homocysteine and oxidized-low-density lipoprotein.

BACKGROUND: Increased levels of homocysteine and oxidized low-density lipoprotein (Ox-LDL) are considered independent risk factors for atherosclerosis. However, no previous study has examined the effects of ethanol-induced increase of homocysteine and Ox-LD on aortic vascular smooth muscle cell (VSMC) proliferation. The aim of the present study was to investigate the relationship between ethanol consumption, increase in homocysteine, Ox-LDL, and aortic VSMC proliferation in rats. METHODS AND RESULTS: To address this issue, 24 male Wistar rats were randomly divided into three groups: control, sham, and ethanol-treated. Homocysteine, Ox-LDL, lipid profile, and aortic VSMC proliferation were assessed after 42 days. The results revealed a concurrent, significant increase in homocysteine and Ox-LDL levels, lipid profile levels, and aortic VSMC proliferation in the ethanol-treated group compared with the control and sham groups. CONCLUSION: Based on these results, we conclude that ethanol apparently exerts aortic VSMC proliferation through increase in homocysteine and Ox-LDL-mediated oxidative stress, which in turn trigger proatherogenic changes in the aortic wall.

Vasoprotective effect of vitamin E: rescue of ethanol-induced atherosclerosis and inflammatory stress in rat vascular wall.

Chronic ethanol consumption increases the incidence of cardiovascular disease. The mechanisms underlying ethanol-induced susceptibility to cardiovascular disease continue to be defined. This study examines the hypothesis that chronic ethanol consumption plausibly induces vascular wall abnormalities via inflammatory reactions. In addition, it intends to find out whether vitamin E inhibits the abnormalities induced by ethanol in rats' vascular wall. Twenty four male Wistar rats were divided into three groups (n=8): Control ©, ethanol (E), and vitamin E treated ethanol (VETE) group. After 6weeks, the aortic and coronary wall changes, vascular endothelial growth factor (VEGF), alpha-1 glycoprotein and haptoglobin amounts in plasma, C-reactive protein levels(CRP), as well as the amount of aortic IL-6 were evaluated. The results revealed the elevation of polymorphonuclear (PMN) leukocyte in the vascular wall, disorganization of endothelium with ballooning of cells, proliferation of vasa-vasorum with an increase in the IL-6, CRP, as well as a decrease in VEGF and an increase in alpha-1 glycoprotein and haptoglobin in the ethanol group compared to the control group. Significant amelioration of aortic and coronary wall changes, along with the restoration of elevated level of IL6, CRP, and the decreased level of VEGF compared to that of the controls were found in vitamin E-treated animals. These findings strongly support the idea that heavy and chronic ethanol consumption initiates atherosclerosis by inflammatory stress, and that these effects can be alleviated by vitamin E as an anti-inflammatory agent.

Decreased blood pressure with a corresponding decrease in adhesive molecules in diabetic rats caused by vitamin E administration.

BACKGROUND: Hypertension is one of the important clinical problems of diabetic cardiovascular disease. The aim of this study was to determine the effect of vitamin E on blood pressure parameters and adhesive molecule amounts in diabetic rats. METHODS: Twenty-four male Wistar rats were divided into three groups (each of n = 8): the controls (C), non-treated diabetic (NTD), and vitamin E treated diabetic (VETD) groups. A single intraperitoneal injection of buffered streptozotocin (60 mg/kg) in cold sodium citrate (pH 4.5) was used to induce diabetes. The VETD group received 300 mg of vitamin E daily intragastrically for 6 weeks. Systolic and diastolic blood pressure, mean arterial pressure, as well as the dicrotic pressure, crest time, systolic and diastolic periods, and plasma levels of intercellular adhesion molecule-1 and E-selectin were measured after 6 weeks. RESULTS: The results revealed that there was a significant increase in systolic and diastolic blood pressures, mean arterial pressure, crest time, systolic duration, and the amount of sICAM-1 and E-selectin in diabetic rats. There was no significant difference in the heart rate or cardiac cyclic duration among the different groups. Significant improvement of blood pressure parameters as well as attenuation of the elevated ICAM-1 and E-selectin amounts was found in the vitamin E treated group. CONCLUSIONS: These findings indicate that vitamin E significantly improved blood pressure elevation in diabetic rats and that these effects could be associated with reducing adhesive molecule and antioxidant properties of vitamin E.