Possible immunological mechanisms in COVID-19 patients with immune thrombocytopenic purpura

Millions of people around the world were, or are still involved with COVID-19 due to infection with SARS-CoV-2. In addition to hallmark symptoms, thrombotic problems, lymphopenia, and thrombocytopenia have also been reported in COVID-19 patients, of which ITP is the most common and occurs in more than one-third of COVID-19 patients. Hyperinflammation, cytokine storms, and generally immune dysregulation in a percentage of patients develop the main consequences of diseases such as ALI, ARDS and multiple organ failure. Some of the important events in the immunopathogenesis of this disease are disruption of T-cell effector differentiation and the destructive role of Th17 lymphocytes, neutrophil function and inflammatory macrophages. NLRP3-inflammasome hyperactivity causes serious dysfunction of innate immune cells and, consequently, T lymphocytes in many inflammatory disorders, most notably in the COVID-19. A closer look at the immunopathogenesis of ITP and COVID-19 brings us to common ground. The purpose of this study was to review and summarize the findings of various studies on the immunopathogenesis of ITP and its possible causes in COVID-19. Finally, enhanced differentiation of Th17 and Th1, the cell death called as pyroptosis, hyperinflammation and dysfunction of inflammatory neutrophils and macrophages, and NLRP3- inflammasome hyperactivity are important factors in the development of thrombocytopenia in patients with COVID-19. Further studies are needed to better understand immunopathogenesis and effective treatments for ITP, especially in inflammatory disorder

Evaluation and Optimization of Lipofectamine 3000 Reagents for Transient Gene Expression in KYSE-30 Esophagus Cancer Cell Line

Background and Aim: Transfection of DNA/RNA sequence into eukaryotic cells has a major effect on scientific studies. Various methods are used to transfer the DNA/RNA sequence into cells, such as lipid-based carriers as the available and easy procedure. Transfection with cationic lipid liposome is introduced as a simple and efficient procedure for monitoring the DNA/RNA sequence through gene function analysis, including fluorescence imaging RNA and protein expression. This study aimed to investigate the transfection efficiency and cell death through GFP expression in human esophageal squamous cell carcinoma (ESCC) cell line KYSE-30 using Lipofectamine 3000 reagent. Methods: The pCDH-513b plasmid DNA was transfected into KYSE-30 cells using Lipofectamine 3000 in different concentrations of the plasmid DNA and reagent. The transfection efficiency was evaluated by fluorescence microscope and flow cytometry analysis to determine the percentage of GFP-expressing cells. Moreover, the viability and death of transfected KYSE-30 cells were evaluated using a trypan blue exclusion assay. Results: The transfection efficiency of KYSE-30 with Lipofectamine 3000 was increased with higher plasmid DNA concentration and a lower amount of Lipofectamine 3000 reagent. The Optimized concentration of 1.5 µg plasmid DNA and volume of one µl of lipofectamine 3000 reagents were identified for 95% transfection efficiency in the KYSE-30 cell line. The viability and death of transfected cells were 43% and 58% after transfection, respectively. Conclusion: The results indicated that Lipofectamine 3000 might not be suitable for transfection in KYSE-30 cells due to increased cell death. *Corresponding Author: Mohammad Reza Abbaszadegan; Email: [email protected] Please cite this article as: Mahmoudian RA, Farshchian M, Abbaszadegan MR. Evaluation and Optimization of Lipofectamine 3000 Reagents for Transient Gene Expression in KYSE-30 Esophagus Cancer Cell Line. Arch Med Lab Sci. 2019;5(4):1-9. https://doi.org/10.22037/amls.v5i4.3108

A review on medicinal plants used for treating ototoxicity and acoustic trauma induced hearing loss

Hearing loss induced by chemotherapy and acoustic trauma is mainly associated with two factors, free radical formation and apoptosis pathway activation. Despite numerous efforts on reducing the effects of these factors, no definite strategy is still determined to interfere with and control these processes. In recent studies, various protective agents, including antioxidants have been used on animal models, to inhibit the formation of free radicals thus improving hearing loss.In this review article we will discuss the role of traditional herbal medicine in treatment of noise/drug induced hearing loss, focusing on medicinal plants’ active substances,as well as their mechanisms of action in reducing or preventing the formation of free radicals thus increasing the rate of survival of cochlea cells. Data have been gathered since year 2000, from scientific publications including the following keywords: deafness, drug toxicity, acute trauma, medicinal herbs and oxidative stress. The study includes all herbs and medicinal plants that have been experimentally used in studies on animal models and clinical trials. The results from these studies indicate the effectiveness of most of these herbs and their active substances through their antioxidative properties. Medicinal plants reported in this review can thus be considered as effective remedies intreating noise/drug induced hearing loss,yet further studies need to be done

Amplification of Tumor Transcripts from Limited Quantity of Esophageal Squamous Cell Carcinoma Tissue Samples

To examine the template-switching technology accompanied by in vitro transcription (the Switch Mechanism At the 5ʹ end of Reverse Transcript) to amplify enough amount of mRNA as input for gene expression experiments. We amplified limited quantity of esophageal squamous cell carcinoma (ESCC) transcripts samples using generated ds cDNA as template and in vitro transcription (IVT) reaction. In addition, the quality and quantity of amplified mRNA were assessed by comparative real-time PCR of genes such as stem cell markers CD44, OCT4 and SNAIL as well as MAGE-A4 as a cancer-testis antigens, and XRCC5 as an underexpressed gene in ESCC.The results obtained from this study demonstrated that optimal amounts of mRNA are generated by template-switching and IVT reaction. Integrity and purity of all RNA samples were assessed. By using this approach, over 10 micrograms of amplified mRNA were generated from 100 ng of starting total RNA. The results of comparative real-time PCR of five genes with different levels of expression illustrated that the expression level of amplified sense RNA was almost similar when compared with non-amplified RNA. Our results clearly showed the usefulness of the T7-based IVT technique for amplification of limited quantity of input total RNA

Overexpression of MiR-183/96/182 Triggers Retina-Like Fate in Human Bone Marrow-Derived Mesenchymal Stem Cells (hBMSCs) in Culture

Retinal degeneration is considered as a condition ensued by different blinding disorders such as retinitis pigmentosa, age-related macular degeneration, and diabetic retinopathy, which can cause loss of photoreceptor cells and also lead to significant vision deficiencies. Although there is no efficient treatment in this domain, transplantation of stem cells has been regarded as a therapeutic approach for retinal degeneration. Thus, the purpose of this study was to analyze the potential of human bone marrow-derived mesenchymal stem cells (hBMSCs) to differentiate into photoreceptor cells via transfection of microRNA (miRNA) in vitro for regenerative medicine purposes. To this end, miR-183/96/182 cluster was transfected into hBMSCs; then, qRT-PCR was performed to measure the expression levels of miR-183/96/182 cluster and some retina-specific neuronal genes such as OTX2, NRL, PKC alpha, and recoverin. CRX and rhodopsin (RHO) levels were also measured through qRT-PCR and immunocytochemistry, and subsequently, cellular change morphology was detected. The findings showed no changes in the morphology of the given cells, and the expression of the neuroretinal genes such as OTX2, NRL, and PKC alpha. Moreover, recoverin was upregulated upon miR-183/-96/-182 overexpression in cultured hBMSCs. Ectopic overexpression of the miR-183 cluster could further increase the expression of CRX and RHO at the messenger RNA (mRNA) and protein levels. Furthermore, the data indicated that the miR-183 cluster could serve as a crucial function in photoreceptor cell differentiation. In fact, miRNAs could be assumed as potential targets to exploit silent neuronal differentiation. Ultimately, it was suggested that in vitro overexpression of miR-183 cluster could trigger reprogramming of the hBMSCs to retinal neuron fate, especially photoreceptor cells. keywords: DIFFERENTIATION; DELETION; CRX; PHOTORECEPTORS; FIBROBLASTS; MATURATION; EPITHELIUM; MICRORNAS; MIR-182; PROTEI

Studying gap junction beta 2-related deafness in Iranian population

Hearing loss is the most common sensory disorder in humans, from every 1000 births, 1 is affected by severe to profound deafness. Many genes are involved in deafness that GJB2 gene is one of the most important ones and encodes the connexin 26 proteins. A mutation called delG35 composes most of mutated alleles of connexin 26 and is also the most common cause of congenital sporadic and hereditary deafness. Open Access Journals (DOAJ), Google Scholar, PubMed (NLM), LISTA (EBSCO), and Web of Science have been searched for literature. GJB2 gene mutations play the most significant role in non-syndromic deafness in Iran. 35delG mutation has a high frequency in most parts of Iran, especially in the North and North-West and is also the most common mutation in GJB2 deaf population of Iran. GJB2 gene mutations in Iran play less important roles compared with other countries in causing deafness; however, so far, it has been introduced as the gene that plays the most significant role in causing deafness gene in Iran. It seems that many other genes and loci play roles in causing deafness in Iran that requires more studies to be conducted. (Cite this article as: Mehri-Ghahfarrokhi A, Hashemzadeh-Chaleshtori M, Shojaeian A, Mahmoudian-Sani MR. Studying gap junction beta 2-related deafness in Iranian population. Otorinolaringol 2017;67:89-95. DOI: 10.23736/S0392-6621.17.02115-4

Ginkgo Biloba in the treatment of tinnitus: An updated literature review

Introduction: Tinnitus is one of the common diseases of the ear that is associated with numerous physical and mental disorders. One of the known mechanisms in the tinnitus area with unknown reason is oxidative events. Based on the prevalence and economic costs and physical- psychological side effects caused by tinnitus and the importance of finding a suitable solution for its prevention and treatment, the need for further studies becomes more obvious in this context. This review article aimed to review studies on the effectiveness of Ginkgo biloba as a medicinal plant on patients with tinnitus. Evidence Acquisitions: Google Scholar, Directory of Open Access Journals (DOAJ), PubMed, LISTA (EBSCO) and Web of Science have been searched. Results: There are many studies on the therapeutic effect of Ginkgo biloba on patients with tinnitus. Most findings are in contrast with each other so that some of studies reported that Ginkgo biloba is effective in the treatment of tinnitus and other studies referred to it as ineffective herbal medicine. Generally, according to the previous studies and the present study, it can mention that the Ginkgo biloba may somewhat improve tinnitus. Conclusion: Since tinnitus is multifactorial, it is recommended to evaluate patients individually based on the cause of tinnitus, treatment formulas, and different doses of Ginkgo biloba at the more extensive level in future studies

A review of medicinal plants for the treatment of earache and tinnitus in Iran

Background and purpose: Despite numerous trials, there has not yet been any definite strategy to reduce replicable long-term tinnitus and earache. Complementary and alternative medical approaches have been used to decrease the symptoms of tinnitus and earache. This study was conducted to report medicinal plants that are used to treat ear disorders, especially earache and tinnitus in different regions of Iran. Evidence Acquisitions: Directory of Open Access Journals (DOAJ), Google Scholar, PubMed, LISTA (EBSCO), Embase, and Web of Science were searched using relevant search terms to retrieve eligible publications. Results: Twenty-three species from sixteen families were used for the treatment of earache and tinnitus in Iran. Plants from families Asteraceae and Lamiaceae were the most commonly used plants for the treatment of earache. Ginkgo biloba was frequently reported for the treatment of tinnitus. Conclusion: This study shows the important role of medicinal plants in the treatment of earache and tinnitus in some regions of Iran. The medicinal plants reported in this review can be considered in treatments for earache and tinnitus if examined more extensively in clinical trials

A review on medicinal plants used for treating ototoxicity and acoustic trauma induced hearing loss

Hearing loss induced by chemotherapy and acoustic trauma is mainly associated with two factors, free radical formation and apoptosis pathway activation. Despite numerous efforts on reducing the effects of these factors, no definite strategy is still determined to interfere with and control these processes. In recent studies, various protective agents, including antioxidants have been used on animal models, to inhibit the formation of free radicals thus improving hearing loss.In this review article we will discuss the role of traditional herbal medicine in treatment of noise/drug induced hearing loss, focusing on medicinal plants’ active substances,as well as their mechanisms of action in reducing or preventing the formation of free radicals thus increasing the rate of survival of cochlea cells. Data have been gathered since year 2000, from scientific publications including the following keywords: deafness, drug toxicity, acute trauma, medicinal herbs and oxidative stress. The study includes all herbs and medicinal plants that have been experimentally used in studies on animal models and clinical trials. The results from these studies indicate the effectiveness of most of these herbs and their active substances through their antioxidative properties. Medicinal plants reported in this review can thus be considered as effective remedies intreating noise/drug induced hearing loss,yet further studies need to be done.publishersversionpublishe