Epidemiological and clinical risk factors related to severe COVID-19 in Iran: a multi-center study.

BACKGROUND: Iran was one of the first countries to be affected by COVID-19. Identifying factors associated with the severity of COVID-19 is effective in disease management. This study investigated the epidemiological and clinical features and factors related to the severity of COVID-19 in one of the less privileged areas in Iran. METHODS: In a multi-center study, all patients admitted to Zahedan University of Medical Sciences hospitals in southeastern Iran were investigated from February 29 to April 31, 2020. Demographic, epidemiological, and clinical data of patients were extracted from medical records. Bivariate and multivariate logistic regression models were used to explore the risk factors associated with the severity of COVID-19. RESULTS: Among the 413 patients, 55.5% were male, and 145 (35.10%) were in a severe condition at admission time. Multivariate analysis showed that the adjusted odds of the disease severity increased in patients with older age (OR 2.27; 95% CI 1.41-3.65), substance abuse (OR 2.49; 95% CI 1.14-5.43), having one underlying disease (OR 1.52; 95% CI 0.90-2.55), having two underlying disease (OR 2.31; 95% CI 1.19-4.50), and having three or more underlying disease (OR 2.60; 95% CI 1.19-5.66). CONCLUSIONS: COVID-19 was more severe in older patients, patients with a history of substance abuse, and patients with the underlying disease. Understanding the factors affecting the disease severity can help the clinical management of COVID-19, especially in less privileged areas where fewer resources are available.

Coding Variants of the FMO3 Gene Are Associated with the Risk of Chronic Kidney Disease: A Case-Control Study.

Background: Chronic kidney disease (CKD) is a global health concern involving roughly one-tenth of developed countries' populations. The flavin-containing dimethylaniline monooxygenase 3 (FMO3) gene encodes an enzyme that catalyzes trimethylamine N-oxide (TMAO), a toxin in CKD sufferers. This preliminary study aims to evaluate the association between coding region variations of FMO3, rs2266782G/A (E158K), rs2266780A/G (E308G), and rs1736557G/A (V257M), and the susceptibility to CKD. Methods: A total of 356 participants were enrolled, including 157 patients diagnosed with CKD and 199 age-matched healthy individuals. Genotyping of FMO3 gene variations was performed via PCR-RFLP and ARMS-PCR methods. Results: Our findings revealed a significant association between rs2266780A/G and rs1736557G/A and CKD under different genetic models. Compared to the GGG haplotype of rs2266782/rs1736557/rs2266780, the GAG, GAA, AAG, and AAA haplotype combinations conferred an increased risk of CKD in our population. Interaction analysis revealed that some genotype combinations, including GA/AA/AA, AA/AA/AA, GA/AA/GA, and GG/AG/AA, dramatically increased CKD risk in the Iranian population. No correlation was found between FMO3 polymorphisms and CKD stages. Discussion: These observations highlight the potential impact of coding variants of the FMO3 gene on the onset of CKD. Further investigations into expanded populations and diverse races are needed to confirm our findings.

Association between angiotensinogen (AGT), angiotensin-converting enzyme (ACE) and angiotensin-II receptor 1 (AGTR1) polymorphisms and COVID-19 infection in the southeast of Iran: a preliminary case-control study.

BACKGROUND: The COVID-19 pandemic remains an emerging public health crisis with serious adverse effects. The disease is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV--2) infection, targeting angiotensin-converting enzyme-2 (ACE2) receptor for cell entry. However, changes in the renin-angiotensin system (RAS) balance alter an individual's susceptibility to COVID-19 infection. We aimed to evaluate the association between AGT rs699 C > T, ACE rs4646994 I/D, and AGTR1 rs5186 C > A variants and the risk of COVID-19 infection and the severity in a sample of the southeast Iranian population. METHODS: A total of 504 subjects, including 258 COVID-19 positives, and 246 healthy controls, were recruited. Genotyping of the ACE gene rs4646994, and AGT rs699, and AGTR1 rs5186 polymorphisms was performed by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP), respectively. RESULTS: Our results showed that the II genotype of ACE rs4646994 and the I allele decreased the risk of COVID-19 infection. Moreover, we found that the TC genotype and C allele of AGT rs699 increased the risk of COVID-19 infection. The AGTR1 rs5186 was not associated with COVID-19 infection. Also, we did not find any association between these polymorphisms and the severity of the disease. However, we found a significantly higher age and prevalence of diabetes and hypertension in patients with severe disease than a non-severe disease. CONCLUSIONS: These findings suggest that ACE rs4646994 and AGT rs699 polymorphisms increase the risk of COVID-19 infection in a southeast Iranian population.

The Effect of the CO2 Fractional Laser or Premarin Vaginal Cream on Improving Sexual Function in Menopausal Women: A Randomized Controlled Trial.

Introduction: Sexual dysfunction is a complex problem in postmenopausal women with a prevalence rate of 68%-86%. This study aimed to evaluate the effect of a fractional CO2 laser or vaginal cream on the improvement of sexual function in menopausal women. Methods: This is a two-group clinical trial study. Postmenopausal women with the inclusion criterion were enrolled and randomly divided into 2 groups of intervention (n=25) and control group (n=25). In the intervention group, CO2 laser therapy was performed every month for three months, and in the control group, Premarin vaginal cream was applied (0.625 mg, 3 nights a week for 3 months. Vaginal health index (VHI) scoring and Female Sexual Function Index (FSFI) questionnaires were completed before and three months after the treatment. Results: The effects of the laser treatment were greater than the Premarin group with respect to improvement in sexual desire, orgasms, sexual satisfaction, less pain during sexual relations, and overall sexual function (P <0.05). Conclusion: It seems that the fractional CO2 laser may be more effective than hormonal therapy in improving sexual function in postmenopausal women.

Levisticum Officinale Extract Triggers Apoptosis and Down-Regulates ZNF703 Gene Expression in Breast Cancer Cell Lines.

BACKGROUND: Studies have shown that zinc finger protein 703 (ZNF703) is overexpressed in breast cancer. Levisticum (L.) officinale is a herbal plant with proven medical characteristics in traditional medicine. The purpose of the present study was to evaluate the effect of hydroalcoholic extract of L. officinale (HELO) on both estrogen receptor-positive (ER+) and -negative (ER-) cell lines (MCF-7 and MDA-MB-468, respectively). METHODS: The anti-proliferative and apoptotic activities of HELO were investigated on both cell lines using MTT and flow-cytometry methods. Real-time PCR was employed to determinate the changes in mRNA expression of the ZNF703 gene. RESULTS: The 50% maximal inhibitory concentrations (IC50s) of HELO on ER+ and ER- cells were 200 and 150 µg/mL after 48 h-treatment. Statistically significant increases in both early and late apoptosis rates were seen in exposed cell lines. ZNF703 expression was less from 4 to 24 h HELO treatment than in untreated cells, and ZNF703 expression was higher in the more invasive MDA-MB-468 cells than in the less invasive MCF-7 cells. Our results demonstrated that HELO induces apoptosis and decreases cell growth in both cell lines. CONCLUSION: Our data suggest that HELO alters the mRNA levels of ZNF703 gene while inducing apoptotic cell death in breast cancer-derived cell lines. The use of ZNF703 suppression can be considered as a beneficial target in breast cancer research.

Trimethylamine-N-oxide, as a risk factor for atherosclerosis, induces stress in J774A.1 murine macrophages.

PURPOSE: Trimethylamine N-oxide (TMAO) is a biomarker for kidney problems. It has also been introduced as a risk factor for atherosclerosis. The classic risk factors for atherosclerosis trigger cellular and humeral immunoreaction in macrophages through induction of heat shock protein expressions and increased levels of GRP94 and HSP70 are associated with increased atherosclerosis risk. The present study evaluated the possible effect(s) of TMAO on the expression of GRP94 and HSP70 at protein levels. METHODS: J774A.1 murine macrophages were treated with different micromolar concentrations of TMAO and 4-phenylbutyric acid (PBA), a chemical chaperone, for 8, 18, 24, and 48h intervals. Tunicamycin was also used as a control for induction of endoplasmic reticulum stress. Western blotting was used to evaluate the expression of GRP94 and HSP70 in macrophages at protein levels. RESULT: Tunicamycin greatly increased protein levels of GRP94. Similarly, but to a lesser extent compared to tunicamycin, TMAO also increased GRP94. In 24h treated cells, only 300µM of TMAO, and in cells treated for 48h, all doses of TMAO produced a significant increase in relative HSP70 protein levels compared to the control. PBA failed to induce any changes in GRP94 or HSP70 protein levels. CONCLUSION: GRP94 and HSP70 are stress-inducible heat shock protein, so the elevation in J774A.1 murine macrophages can clearly define cells under stress and elucidate the contribution of stress induced by TMAO that may have a part in the abnormal activation of macrophages involved in foam cell formation.

Different Expressions of Specific Transcription Factors of Th1 (T-bet) and Th2 cells (GATA-3) by Peripheral Blood Mononuclear Cells From Patients With Multiple Sclerosis.

INTRODUCTION: Multiple Sclerosis (MS) is an inflammatory disorder caused by self-reactive Th1 lymphocytes, while Th2 cells may confer protection. The Th1 and Th2 cell differentiation are regulated by specific transcription factors, especially T-bet and GATA-3, respectively. This investigation aimed to measure the T-bet and GATA-3 expression by Peripheral Blood Mononuclear Cells (PBMCs) obtained from MS patients after specific and non-specific in vitro stimulation. METHODS: The PBMCs were separated from 22 patients with MS and 20 healthy individuals. They were cultured at 37°C for 24 h in the absence of a stimulator or in the presence of Myelin oligodendrocyte Glycoprotein (MOG) or Phytohemagglutinin (PHA) at a concentration of 10 µg/mL. Then the T-bet and GATA-3 expression was measured by real time-PCR. RESULTS: The T-bet expression was enhanced, while the GATA-3 expression diminished. Therefore the expression of T-bet/GATA-3 ratio diminished in not-stimulated, MOG-stimulated and PHA-stimulated PBMCs from MS patients compared with equal cultures from the healthy individuals (P<0.01, P<0.01 and P<0.01, for T-bet; P<0.03, P<0.01 and P<0.02, for GATA-3; P<0.01, P<0.001 and P<0.01 for T-bet/GATA-3 ratio, respectively). The not-stimulated, MOG-stimulated, and PHA-stimulated PBMCs from men with MS expressed higher amounts of GATA-3 than equal cells from MS women (P<0.05, P<0.05 and P<0.01, respectively). CONCLUSION: These results probably indicate an imbalance in Th1/Th2 cells in the level of transcription factors with a tendency toward Th1 cells in MS. The clinical utilization of the transcription factors as novel biomarkers of MS should be evaluated in further studies.

Altered Expression of Specific Transcription Factors of Th17 (RORγt, RORα) and Treg Lymphocytes (FOXP3) by Peripheral Blood Mononuclear Cells from Patients with Multiple Sclerosis.

The imbalance in Th17/Treg cell-related responses plays an important role in the pathogenesis of multiple sclerosis (MS). The development of Th17- and Treg cells is regulated by specific transcription factors-RORγt and RORα-and FOXP3, respectively. The aim was to determine the expression of RORγt, RORα, and FOXP3 in peripheral blood mononuclear cells (PBMCs) from MS patients following in vitro stimulation. The PBMCs from 22 MS patients and 20 healthy subjects were cultured in the presence of 10 µg/ml MOG, 10 µg/ml PHA, or without stimulation. The PBMCs were incubated at 37 °C for 24 h, and then the messenger RNA (mRNA) expression of RORγt, RORα, and FOXP3 was determined by real-time PCR. The expression of RORγt and RORα was increased in non-stimulated, MOG-stimulated, and PHA-stimulated PBMCs from MS patients in comparison with same cultures from the healthy group (P < 0.01, P < 0.01, and P < 0.02 for RORγt; P < 0.001, P < 0.001, and P < 0.05, for RORα, respectively). The FOXP3 expression in non-stimulated PBMCs from MS patients was significantly lower than that in equal culture from healthy subjects (P < 0.05). There were no significant differences between healthy subjects and MS patients regarding the expression of FOXP3 mRNA by MOG-stimulated and PHA-stimulated PBMCs. These results showed an imbalance in Th17/Treg cells at transcription factor levels with a deviation toward Th17 cell in MS. The correction of Th17/Treg balance at transcription levels should be considered to design novel therapeutic strategies for MS treatment.