The Future of Iran's Population: Balancing Aging Trends and Fertility Rates 2 2 As current population growth trends persist, Iran's population is projected to reach 42 million by the year 1480, marking a significant transition from a predominantly youthful demographic to an increasingly aging society. While European countries face similar demographic challenges, their population decline is occurring gradually. In contrast, Iran's declining youth population presents a more urgent concern. Presently, there are approximately 5 million couples, representing 10 million individuals of marriageable age, who remain unmarried. It is imperative to provide these individuals with essential resources, including employment opportunities and housing, to facilitate family formation (1). Another critical issue is the prevailing trend of having only one child. As parents age, their children face the difficult choice between establishing their own lives and providing care for their aging parents, a dilemma that poses significant implications for future demographic stability. In approximately 20 years, the rates of birth and death in Iran are expected to equalize, resulting in zero population growth. Subsequently, the number of deaths is projected to exceed births, leading to a decline in the population to below 50 million by the year 1480 (1). Current demographic data indicate that approximately 24% of Iran's population of 85 million consists of children and adolescents under the age of 15. Additionally, 25% are young individuals aged 15 to 29, while 44% are middle-aged, and the remaining population comprises elderly individuals over 65. This demographic structure highlights a predominance of middle-aged individuals. Alarmingly, road traffic accidents claim the lives of approximately 20,000 individuals annually, a significant proportion of whom are productive young people. Urgent preventive measures are necessary to address this issue, including improvements in road conditions, driving culture, and vehicle safety. The car industry, while contributing to road fatalities, also exacerbates environmental pollution through inefficient fuel combustion, positioning it as a potential barrier to population growth. Furthermore, the interval between marriage and the birth of the first child currently spans 4.5 to 5 years, a gap that must be addressed. Delayed marriage and extended intervals between childbirth can diminish the desire for larger families. The average age of marriage among Iranian youth, particularly in provinces where fertility rates have fallen below 1.5, has been on the rise. To counteract these trends, targeted population interventions should be implemented in provinces with fertility rates below 1.5, promoting childbearing through practical measures rather than mere rhetoric. This responsibility extends beyond the Ministry of Health to encompass the entire government. While Level 2 and 3 infertility treatment centers have been established across all provinces, providing essential services to infertile couples, the number of such centers exceeds the number of provincial health facilities. Addressing demographic challenges requires a multifaceted approach that considers economic, social, and cultural factors alongside medical interventions. Over the past 35 years, Iran has made significant strides in infertility treatment, emerging as a regional leader in this field, with health insurance covering nearly 100% of infertility treatment costs. In conclusion, a comprehensive and coordinated policy response is essential to address the demographic challenges facing Iran. By fostering an environment conducive to family formation and addressing the underlying social determinants of fertility, we can work towards a sustainable demographic future. 82 82 Ladan Kashani Infertility Ward, Arash Hospital, Tehran University of Medical Sciences Infertility Ward, Arash Hospital, Tehran University of Medical Sciences Iran Shahin Akhondzadeh Editorial 60615.pdf Joulaei H, Morshed-Behbahani B, Ghadimi P, Nadimi Parashkouhi S, Mansoori Y. Contentious population policy-making and its consequences: a health policy analysis. Int J Equity Health 2025;24(1):96.##

Revolutionary Regeneration Therapy Utilizing Dental Stem Cells and State-of-the-Art Nanotechnology Devices to Heal Injured Teeth and Tissues 2 2 Regenerative medicine is a field of pharmacy and medicine that focuses on stem cells and other methods such as nanoscience and biotechnology to stimulate the body's natural regenerative processes and repairing damaged tissues and organs to improve function and reduce pain. In this review article, focus is on Dental Stem Cells (DSC) and other cells regeneration in human body. The appropriateness of tissue-engineered therapies relying on the multipotent regenerative abilities of DSC is accompanied by significant challenges, as growth factors and epigenetic components are crucial for preserving their multipotency while being susceptible to a range of natural and environmental factors. Current evidence highlights the positive outcomes associated with select regenerative therapies; nevertheless, to provide further support, additional data must be gathered through standardized therapies and further studies. Organoids (3D cell culture) and nano scaffolds are also being explored as potential tools for regenerative therapies. Understanding the mechanisms that determine the behavior of these cells and how they interact will enable future generation therapies. Demonstrating promise, cell therapy is an alternative approach within regenerative medicine. Developmental factors like extracellular vesicle production are thought to mediate the regenerative response through paracrine effects in cell therapy, which is widely recognized. 83 97 Mohammad Balfaki Erfan Salimi Tib University of Baku Tib University of Baku Azerbaijan Cell cultureImmunomodulationOrganoidsRegenerative medicineScaffoldsSignal pathwaysStem cell research 60606.pdf Egger A, Tomic-Canic M, Tosti A. 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Monocytes and Macrophages as Unique Cellular Compartments Governing Non-Alcoholic Fatty Liver Disease and Inflammation 2 2 Non-Alcoholic Fatty Liver Disease (NAFLD) is a spectrum of liver diseases from simple steatosis to the most severe form of hepatocellular carcinoma. Liver injuries resulting from various factors, including viral infections, alcohol consumption, and metabolic disorders, trigger the activation of resident immune cells and the recruitment of circulating immune cells to the liver. This chronic inflammatory environment leads to tissue damage and the progression of liver fibrosis. Macrophages are highly versatile immune cells that play a dual role in fibrosis: they contribute to the progression of fibrosis (M1 and Ly6chigh macrophages) and its resolution (M2 and Ly6clow macrophages). M1 macrophages and those with high surface expression of Ly6C exhibit pro-inflammatory characteristics, while M2 macrophages and myeloid cells with low expression of Ly6C mitigate inflammation and inhibit fibrosis progression. Environmental stimuli influence the complex mechanisms hepatic macrophages regulate the fibrosis they encounter. Kupffer cells initiate the inflammatory cascade and recruit monocyte-derived macrophages, which modulate the propagation of fibrosis and promote fibrinolysis. Additionally, hepatic macrophages interact with other cell types through exosomes, facilitating the transfer of cellular components that influence the outcome of liver fibrosis. In this review, the critical role of macrophages in inflammation-induced fibrosis and tissue restoration is discussed. 97 105 Ghazale Hemmatian Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Iran Davoud Rostamzadeh Medicinal Plants Research Center, Yasuj University of Medical Sciences Medicinal Plants Research Center, Yasuj University of Medical Sciences Iran Kaveh Baghaei Gastroenterology and Liver Disease Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Gastroenterology and Liver Disease Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Iran Mahdi Shabani CarcinomaExosomesFibrinolysisHepatocellularKupffer cellsLiver cirrhosisLiver neoplasmsNon-alcoholic fatty liver disease 60607.pdf Matteoni CA, Younossi ZM, Gramlich T, Boparai N, Liu YC, McCullough AJ. 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Investigation of Anti-Cancerous Effects of L. casei –ATCC-393 and L. rhamnosus-GG on Apoptosis and Cell Cycle of B- CPAP Thyroid Cancer Cell line in Comparison to Fibroblast Cell Line 2 2 Background: Thyroid cancer is the most common type of cancer affecting the endocrine system. The main treatment approaches consist of surgical procedures and radioiodine therapy. Recently, there has been a heightened interest in investigating alternative treatment options, including probiotics, which could potentially minimize toxicity. Consequently, there is a growing imperative for research aimed at investigating the potential role of probiotics in the management of cancer. Methods: The B-CPAP cell line was maintained in culture and tested with different dilutions of two bacterial strains. Toxicity evaluations were performed using the MTT assay to identify appropriate concentrations. mRNA was extracted and analyzed via real-time PCR to measure the expression levels of the Bcl-2, Bax, and P53 genes. Furthermore, changes in the cell cycle and the induction of apoptosis were examined using flow cytometry. Results: The supernatant derived from Lacticaseibacillus casei (L. casei) – ATCC-393 and Lacticaseibacillus rhamnosus (L. rhamnosus)-GG demonstrated a significant inhibitory effect on the growth of B-CPAP cancer cells. The findings indicated that the combination produced a more pronounced anti-cancer effect by enhancing the expression of pro-apoptotic genes while reducing Bcl-2 gene expression in B-CPAP cells. Conclusion: The findings indicated a notable change in the expression of genes associated with apoptosis and modifications in the cell cycle. This implies that probiotics may enhance the efficacy of chemotherapy in treating thyroid cancer. In particular, L. rhamnosus and L. casei may play a beneficial role in the therapeutic process. Further research is required to investigate the direct impact of probiotics on thyroid function. 106 113 Maryam Honardoost Endocrine Research Center, Institute of Endocrinology and Metabolism, Iran University of Medical SciencesCardio-Oncology Research Center, Rajaie Cardiovascular Research Institute, Iran University of Medical Sciences Endocrine Research Center, Institute of Endocrinology and Metabolism, Iran University of Medical SciencesCardio-Oncology Research Center, Rajaie Cardiovascular Research Institute, Iran University of Medical Sciences IranIran Fatemeh Soleimanifar Solat Eslami Department of Medical Biotechnology, School of Medicine, Alborz University of Medical Sciences Department of Medical Biotechnology, School of Medicine, Alborz University of Medical Sciences Iran Sara Cheraghi Endocrine Research Center, Institute of Endocrinology and Metabolism, Iran University of Medical Sciences Endocrine Research Center, Institute of Endocrinology and Metabolism, Iran University of Medical Sciences Iran Mohammad Ebrahim Khamseh Endocrine Research Center, Institute of Endocrinology and Metabolism, Iran University of Medical Sciences Endocrine Research Center, Institute of Endocrinology and Metabolism, Iran University of Medical Sciences Iran Maryam Darvish Department of Medical Biotechnology, Faculty of Medicine, Arak University of Medical Science Department of Medical Biotechnology, Faculty of Medicine, Arak University of Medical Science Iran Hamed Haddad Kashani Anatomical Sciences Research Center, Institute for Basic Sciences, Kashan University of Medical Sciences Anatomical Sciences Research Center, Institute for Basic Sciences, Kashan University of Medical Sciences Iran Apoptosisbcl-2-associated X proteinCell cycleCell lineFlow cytometryGene expressionIodine radioisotopesProbioticsReal time polymerase chain reactionThyroid neoplasms 60608.pdf Yuan L, Yang P, Wei G, Hu X, Chen S, Lu J, et al. Tumor microbiome diversity influences papillary thyroid cancer invasion. Commun Biol 2022 Aug 24;5(1):864.##Zhang J, Zhang F, Zhao C, Xu Q, Liang C, Yang Y, et al. Dysbiosis of the gut microbiome is associated with thyroid cancer and thyroid nodules and correlated with clinical index of thyroid function. Endocrine 2019;64(3):564-74.##Jašarević E, Morrison KE, Bale TL. Sex differences in the gut microbiome-brain axis across the lifespan. Philos Trans R Soc Lond B Biol Sci 2016 Feb 19;371(1688):20150122.##Jiang, W. Lu G, Gao D, Lv Z, Li D. The relationships between the gut microbiota and its metabolites with thyroid diseases. Front Endocrinol (Lausanne). 2022 Aug 18;13:943408. 943408.##Mojibi P, F Tafvizi, M. Bikhof Torbati. Cell-bound Exopolysaccharide Extract from Indigenous Probiotic Bacteria Induce Apoptosis in HT-29 cell-line. Iran J Pathol 2019;14(1):41-51.##Dehghani, N., F. Tafvizi, and P. Jafari, Cell cycle arrest and anti-cancer potential of probiotic Lactobacillus rhamnosus against HT-29 cancer cells. Bioimpacts, 2021. 11(4): p. 245-252.##de Moreno de Leblanc A and Perdigón G. The application of probiotic fermented milks in cancer and intestinal inflammation. Proc Nutr Soc 2010;69(3):421-8.##Karimi Ardestani S, Tafvizi F, Tajabadi Ebrahimi M. Heat-killed probiotic bacteria induce apoptosis of HT-29 human colon adenocarcinoma cell line via the regulation of Bax/Bcl2 and caspases pathway. Hum Exp Toxicol 2019;38(9):1069-81.##Reid G, Jass J, Sebulsky MT, McCormick JK. Potential uses of probiotics in clinical practice. Clin Microbiol Rev 2003;16(4):658-72.##Sugimura N, Li Q, Chu ESH, Lau HCH, Fong W, Liu W, et al. Lactobacillus gallinarum modulates the gut microbiota and produces anti-cancer metabolites to protect against colorectal tumourigenesis. Gut 2021;71(10):2011-21.##Soltan Dallal MM, Mojarrad M, Baghbani F, Raoofian R, Mardaneh J, Salehipour Z.Effects of probiotic Lactobacillus acidophilus and Lactobacillus casei on colorectal tumor cells activity (CaCo-2). Arch Iran Med 2015;18(3):167-72.##Kahouli I, Malhotra M , Alaoui-Jamali MA, and Prakash S. In-vitro characterization of the anti-cancer activity of the probiotic bacterium Lactobacillus fermentum NCIMB 5221 and potential against colorectal cancer. J Cancer Sci Ther 2015;7(7): 224-35.##Er S, Koparal AT, Kivanc M. Cytotoxic effects of various lactic acid bacteria on Caco-2 cells. Turkish J Biol 2015;39(1):23-30.##Kun Y, Xiaodong W, Haijun W, Xiazi N, Dai Q. Exploring the oral‐gut microbiota during thyroid cancer: Factors affecting the thyroid functions and cancer development. Food Sci Nutr 2023;11(10):5657-74.##Hou J, Tang Y, Chen Y, Chen D. The Role of the Microbiota in Graves' Disease and Graves' Orbitopathy. Front Cell Infect Microbiol 2021 Dec 22;11:739707.e 739707.##Adeela Yasmin, Sadiq Butt M, Afzaal M, van Baak M, Nadeem MTMuhammad Zia Shahid Prebiotics, gut microbiota and metabolic risks: Unveiling the relationship. Journal of functional foods 2015;17:189-201.##

Recombinase Polymerase Amplification (RPA)-ELISA as an Isothermal Molecular POCT Method for Bacterial Respiratory Infection Diagnosis 2 2 Background: Acute Respiratory Infections (ARIs) are a leading cause of childhood mortality worldwide, especially in African and Southeast Asian countries. Point of Care Test (POCT) techniques provide faster diagnoses compared to conventional or real-time PCR methods. Recombinase Polymerase Amplification (RPA) offers rapid on-site detection of these infections. Coupling RPA with Enzyme-Linked Immunosorbent Assay (ELISA) (RPA-ELISA) creates a cost-effective alternative, ideal for clinical applications. This study evaluates RPA-ELISA as a rapid diagnostic tool for bacterial respiratory infections. Methods: From 11 August 2022 to 9 February 2023, respiratory samples were collected and processed using culture methods, biochemical tests, real-time PCR, and RPA assays. The RPA reactions were conducted at 39°C for 30 min, and ELISA was used for detection. Statistical analyses focused on sensitivity, specificity, Positive Predictive Values (PPV), and Negative Predictive Values (NPV). Results: Forty-two respiratory samples, were collected in this period of which 10 samples showed no growth, and 32 tested positive. Among these positive samples, 15 isolates (35.7%) were identified as Klebsiella pneumoniae (K. pneumoniae), 14 isolates (33.3%) as Streptococcus pneumoniae (S. pneumoniae), and 3 isolates (7.1%) as Moraxella catarrhalis (M. catarrhalis). RPA-ELISA demonstrated 100% sensitivity for all pathogens, comparable to or better than RT-PCR, but had slightly lower specificity and PPV. RT-PCR achieved 100% specificity and PPV for all pathogens, indicating higher accuracy; yet, RPA-ELISA's sensitivity points to its effectiveness as a rapid screening tool. Conclusion: RPA-ELISA is significantly faster than real-time PCR and culture methods. Its ease of use makes it suitable for on-site diagnoses in resource-limited environments. Limitations include a small sample size for certain bacteria and the necessity for further validation in varied clinical contexts. 114 121 Reza Azizian Pediatric Infectious Diseases Research Center (PIDRC), Tehran University of Medical Sciences (TUMS)Biomedical Innovation and Start-up Student Association, Tehran University of Medical Sciences (TUMS) Pediatric Infectious Diseases Research Center (PIDRC), Tehran University of Medical Sciences (TUMS)Biomedical Innovation and Start-up Student Association, Tehran University of Medical Sciences (TUMS) IranIran Erfaneh Jafari Pediatric Infectious Diseases Research Center (PIDRC), Tehran University of Medical Sciences (TUMS)Biomedical Innovation and Start-up Student Association, Tehran University of Medical Sciences (TUMS) Pediatric Infectious Diseases Research Center (PIDRC), Tehran University of Medical Sciences (TUMS)Biomedical Innovation and Start-up Student Association, Tehran University of Medical Sciences (TUMS) IranIran Babak Pourakabri Setareh Mamishi Pediatric Infectious Diseases Research Center (PIDRC), Tehran University of Medical Sciences (TUMS) Pediatric Infectious Diseases Research Center (PIDRC), Tehran University of Medical Sciences (TUMS) Iran Reihaneh Hosseinpour Sadeghi Pediatric Infectious Diseases Research Center (PIDRC), Tehran University of Medical Sciences (TUMS) Pediatric Infectious Diseases Research Center (PIDRC), Tehran University of Medical Sciences (TUMS) Iran Maryam Sotoudeh Anvari Department of Pathology, School of Medicine, Children’s Medical Center, Tehran University of Medical Sciences (TUMS) Department of Pathology, School of Medicine, Children’s Medical Center, Tehran University of Medical Sciences (TUMS) Iran Isothermal methodsPoint of Care Test (POCT)Respiratory infectionsRPARPA-ELISA 60609.pdf Jin X, Ren J, Li R, Gao Y, Zhang H, Li J, et al. Global burden of upper respiratory infections in 204 countries and territories, from 1990 to 2019. EClinicalMedicine 2021;37:100986.##Le Nguyen HK, Nguyen SV, Nguyen AP, Hoang PMV, Le TT, Nguyen TC, et al. Surveillance of severe acute respiratory infection (SARI) for hospitalized patients in Northern Vietnam, 2011–2014. Jpn J Infect Dis 2017;70(5):522-7.##Troeger C, Blacker B, Khalil IA, Rao PC, Cao J, Zimsen SR, et al. Estimates of the global, regional, and national morbidity, mortality, and aetiologies of lower respiratory infections in 195 countries, 1990–2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet Infect Dis 2018;18(11):1191-210.##Walker CLF, Rudan I, Liu L, Nair H, Theodoratou E, Bhutta ZA, et al. Global burden of childhood pneumonia and diarrhoea. Lancet 2013;381(9875):1405-16. ##Mohammadi G, Ghorbani R, Khosravifar S, Babakhanian M. Medical Causes of Death in Iranian Children Aged 1-59 Months: A Systematic Review and Meta-analysis. 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Rapid detection of HIV-1 proviral DNA for early infant diagnosis using recombinase polymerase amplification. MBio 2013;4(2): e00135-13.##Abdeldaim GM, Strålin K, Olcén P, Blomberg J, Mölling P, Herrmann B. Quantitative fucK gene polymerase chain reaction on sputum and nasopharyngeal secretions to detect Haemophilus influenzae pneumonia. Diagn Microbiol Infect Dis 2013;76(2):141-6.##Hartman LJ, Selby EB, Whitehouse CA, Coyne SR, Jaissle JG, Twenhafel NA, et al. Rapid real-time PCR assays for detection of Klebsiella pneumoniae with the rmpA or magA genes associated with the hypermucoviscosity phenotype: screening of nonhuman primates. J Mol Diagn 2009;11(5):464-71.##Lee CS, Wetzel K, Buckley T, Wozniak D, Lee J. Rapid and sensitive detection of Pseudomonas aeruginosa in chlorinated water and aerosols targeting gyrB gene using real‐time PCR. J Appl Microbiol 2011;111(4):893-903.##Liao S, Wang L, Ji X, Chen J, Li Q, Ma L. Simultaneous detection of 15 respiratory pathogens with a fluorescence probe melting curve analysis-based multiplex real-time PCR assay. Int J Mol Epidemiol Genet 2019;10(2):29.##Lim HJ, Kang E-R, Park MY, Kim BK, Kim MJ, Jung S, et al. Development of a multiplex real-time PCR assay for the simultaneous detection of four bacterial pathogens causing pneumonia. PLoS One 2021;16(6):e0253402.##Lobato IM, O'Sullivan CK. Recombinase polymerase amplification: Basics, applications and recent advances. Trends Analyt Chem 2018;98:19-35. ##Heeroma AJ, Gwenin C. Development of solid-phase rpa on a lateral flow device for the detection of pathogens related to sepsis. Sensors 2020;20(15):4182.##Tomar S, Lavickova B, Guiducci C. Recombinase polymerase amplification in minimally buffered conditions. Biosens Bioelectron 2022;198:113802.##Santiago-Felipe S, Tortajada-Genaro LA, Puchades R, Maquieira A. Recombinase polymerase and enzyme-linked imMunosorbent assay as a DNA amplification-detection strategy for food analysis. Analytica Chim Acta 2014;811:81-7.##Tan M, Liang L, Liao C, Zhou Z, Long S, Yi X, et al. A rapid and ultra-sensitive dual readout platform for Klebsiella pneumoniae detection based on RPA-CRISPR/Cas12a. Front Cell Infect Microbiol 2024;14:1362513.##Fu J, Mo R, Li Z, Xu S, Cheng X, Lu B, et al. An extraction-free one-pot assay for rapid detection of Klebsiella pneumoniae by combining RPA and CRISPR/Cas12a. Biosens Bioelectron 2025;267:116740.##Wang F, Wang Y, Liu X, Wang L, Wang K, Xu C, et al. Rapid, simple, and highly specific detection of Streptococcus pneumoniae with visualized recombinase polymerase amplification. Front Cell Infect Microbiol 2022;12:878881.##Yu L, Xiao F, Peng B, Jia N, Fu J, Chen M, et al. Rapid, sensitive and highly specific diagnosis of Moraxella catarrhalis by recombinase polymerase amplification-based biosensor and fluorescence detection. 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Optimization of RfxCas13d Expression in Escherichia coli Host using Response Surface Methodology 2 2 Background: RfxCas13d, a key member of the Cas13 family, plays a vital role in CRISPR-based diagnostics for RNA sequence detection and gene silencing. This study aimed to enhance RfxCas13d expression by optimizing key parameters using Response Surface Methodology (RSM). Methods: The plasmid pET28b-RfxCas13d-His (Addgene 141322) was introduced into BL21 (DE3) and Rosetta™ (DE3) strains. Initial expression tests were conducted, followed by RSM-guided optimization of factors such as isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration, temperature, cell density at induction, and induction time in BL21 (DE3). Protein expression levels were quantified using ImageJ and AlphaEaseFC software to analyze band intensities. Results: BL21 (DE3) was selected for further optimization based on preliminary results. Analysis of 26 RSM-designed experiments revealed that temperature, induction time, IPTG concentration, and their interactions significantly influenced RfxCas13d expression. Optimal conditions were identified as 0.25 mM IPTG, an OD600 nm of 0.8 at induction, 37°C, and Overnight (ON) of induction. The regression model exhibited high accuracy, with a correlation coefficient of 0.97 and a p-value less than 0.05, confirming a strong linear relationship between predicted and observed values. Conclusion: This study highlights the significant impact of the four optimized factors on RfxCas13d expression. Under optimized conditions, a soluble protein concentration of 3.6 mg/100 ml cell culture was achieved after purification. It represents the first application of RSM for optimizing RfxCas13d expression, providing a foundation for further refinement of expression conditions. Continued use of RSM in future research will enhance the efficiency of RfxCas13d production for diagnostic and therapeutic applications. 122 130 Sepideh Abbaszadeh Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Hamadan University of Medical SciencesResearch Center for Molecular Medicine, Institute of Cancer, Hamadan University of Medical Sciences Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Hamadan University of Medical SciencesResearch Center for Molecular Medicine, Institute of Cancer, Hamadan University of Medical Sciences IranIran Shahin Eghbalsaied Research Center for Molecular Medicine, Institute of Cancer, Hamadan University of Medical Sciences Research Center for Molecular Medicine, Institute of Cancer, Hamadan University of Medical Sciences Iran Meysam Soleimani Department of Pharmaceutical Biotechnology, School of Pharmacy, Hamadan University of Medical Sciences Department of Pharmaceutical Biotechnology, School of Pharmacy, Hamadan University of Medical Sciences Iran Sadegh Khazalpour Department of Analytical Chemistry, Faculty of Chemistry and Petroleum Science, Bu-Ali Sina University Department of Analytical Chemistry, Faculty of Chemistry and Petroleum Science, Bu-Ali Sina University Iran Saeid Afshar Cancer Research Center, Institute of Cancer, Hamadan University of Medical Sciences Cancer Research Center, Institute of Cancer, Hamadan University of Medical Sciences Iran Base sequenceCRISPR-associated proteinsEscherichia coliProtein biosynthesis 60610.pdf Makarova KS, Aravind L, Wolf YI, Koonin EV. 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Nat Rev Microbiol 2011;9(6):467-77.##Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung J, et al. Nucleic acid detection with CRISPR-Cas13a/C2c2. Science 2017;356(6336):438-42.##Heo W, Lee K, Park S, Hyun K-A, Jung H-I. Electrochemical biosensor for nucleic acid amplification-free and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA via CRISPR/Cas13a trans-cleavage reaction. Biosens Bioelectron 2022;201:113960.##Yu J, Shin J, Yu J, Kim J, Yu D, Heo WD. Programmable RNA base editing with photoactivatable CRISPR-Cas13. Nat Commun 2024;15(1):673.##Huynh N, Depner N, Larson R, King-Jones K. A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila. Genome Biol 2020;21(1):279.##Makarova KS, Wolf YI, Koonin EV. Classification and nomenclature of CRISPR-Cas systems: where from here? CRISPR J 2018;1(5):325-36.##Kellner MJ, Koob JG, Gootenberg JS, Abudayyeh OO, Zhang F. SHERLOCK: nucleic acid detection with CRISPR nucleases. Nat Protoc 2019;14(10):2986-3012.##Sheng Y, Zhang T, Zhang S, Johnston M, Zheng X, Shan Y, et al. A CRISPR/Cas13a-powered catalytic electrochemical biosensor for successive and highly sensitive RNA diagnostics. Biosens Bioelectron 2021;178:113027.##Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD. Transcriptome engineering with RNA-targeting type VI-D CRISPR effectors. Cell 2018;173(3):665-76. e14.##Yan WX, Chong S, Zhang H, Makarova KS, Koonin EV, Cheng DR, et al. Cas13d is a compact RNA-targeting type VI CRISPR effector positively modulated by a WYL-domain-containing accessory protein. Mol Cell 2018;70(2):327-39. e5.##Wei J, Lotfy P, Faizi K, Baungaard S, Gibson E, Wang E, et al. Deep learning and CRISPR-Cas13d ortholog discovery for optimized RNA targeting. Cell Syst 2023;14(12):1087-102. e13.##Manoochehri H, Asadi S, Tanzadehpanah H, Sheykhhasan M, Ghorbani M. CDC25A is strongly associated with colorectal cancer stem cells and poor clinical outcome of patients. Gene Reports 2021;25:101415.##Sheykhhasan M, Ahmadyousefi Y, Seyedebrahimi R, Tanzadehpanah H, Manoochehri H, Dama P, et al. DLX6-AS1: a putative lncRNA candidate in multiple human cancers. Expert Rev Mol Med 2021;23:e17.##Soulari RN, Basafa M, Rajabibazl M, Hashemi A. Effective strategies to overcome the insolubility of recombinant ScFv antibody against EpCAM extracellular domain in E. coli. International Journal of Peptide Research and Therapeutics 2020;26(4):2465-74.##Boyle DM, Buckley JJ, Johnson GV, Rathore A, Gustafson ME. Use of the design‐of‐experiments approach for the development of a refolding technology for progenipoietin‐1, a recombinant human cytokine fusion protein from Escherichia coli inclusion bodies. Biotechnol Appl Biochem 2009;54(2):85-92.##Emamipour N, Vossoughi M, Mahboudi F, Golkar M, Fard-Esfahani P. Soluble expression of IGF1 fused to DsbA in SHuffle™ T7 strain: optimization of expression and purification by Box-Behnken design. Appl Microbiol Biotechnol 2019;103:3393-406.##Aghaeepoor M, Akbarzadeh A, Kobarfard F, Shabani AA, Dehnavi E, Aval SJ, et al. Optimization and high level production of recombinant synthetic Streptokinase in E. coli using Response Surface Methodology. Iran J Pharm Res 2019;18(2):961-73.##Ghasemi N, Bandehpour M, Ranjbari J. Optimization of key factors in serum free medium for production of human recombinant GM-CSF using response surface methodology. I Iran J Pharm Res 2019;18(Suppl1):146.##Bezerra MA, Santelli RE, Oliveira EP, Villar LS, Escaleira LA. Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta 2008;76(5):965-77.##Kinna A, Tolner B, Rota EM, Titchener‐Hooker N, Nesbeth D, Chester K. IMAC capture of recombinant protein from unclarified mammalian cell feed streams. Biotechnol Bioeng 2016;113(1):130-40.##Morowvat MH, Babaeipour V, Rajabi-Memari H, Vahidi H, Maghsoudi N. Overexpression of recombinant human beta interferon (rhINF-β) in periplasmic space of Escherichia coli. Iran J Pharm Res 2014;13(Suppl):151-60.##Papaneophytou CP, Kontopidis G. Statistical approaches to maximize recombinant protein expression in Escherichia coli: a general review. Protein Expr Purif 2014;94:22-32.##Rostami N, Goharrizi LY. Cloning, expression, and purification of the human synthetic survivin protein in Escherichia Coli using response surface methodology (RSM). Mol Biotechnol 2023;65(3):326-36.##Sambrook J. Molecular cloning: A laboratory manual, Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. 9. (No Title). 1989;14:23.##Chen W-H, Uribe MC, Kwon EE, Lin K-YA, Park Y-K, Ding L, et al. 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The Pro-Apoptosis Induction of Teucrium persicum Ethyl Acetate Extract on MCF-7 Cells: An In Vitro Study 2 2 Background: Teucrium persicum (T. persicum) is a well-known Iranian endemic plant that grows in the southern regions of Iran. It is used as a tea to treat abdominal pains, hyperlipidemia, and diabetes in traditional Iranian medicine. It has been previously found that the methanolic extract of T. persicum exerts significant cytotoxicity and inhibitory effects on different cancer cells. This study aimed to investigate the effects of ethyl acetate extract of T. persicum on MCF-7 cells. Methods: The experiments included MTT, DAPI staining, and investigating the expression of BAX and BCL2 genes. The extract had a significant cytotoxic effect on MCF-7 cells, with an IC50 value of 50 µg/ml for 48 hr. Results: DAPI staining assays showed that the extract induced morphological changes, chromatin condensation, and nuclear fragmentation. Additionally, the ethyl acetate extract induced the expression of BAX and down-regulated BCL2 genes. Conclusion: These findings suggest that T. persicum has strong cytotoxic properties and warrants further investigation. 131 135 Mehdi Moeil Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran Iran Majid Tafrihi Ehsan Nazifi Department of Biology, Faculty of Science, University of Mazandaran Department of Biology, Faculty of Science, University of Mazandaran Iran Maryam Radfar Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran Iran ApoptosisBreast cancerCytotoxicityGene expressionTeucrium persicum 60611.pdf Mattiuzzi C, Lippi G. Current cancer epidemiology. J Epidemiol Glob Health 2019; 9(4):217-22.##Dehelean CA, Marcovici I, Soica C, Mioc M, Coricovac D, Iurciuc S, et al. Plant-derived anticancer compounds as new perspectives in drug discovery and alternative therapy. Molecules. 2021;26(14):1109.##Asma ST, Acaroz U, Imre K, Morar A, Shah SRA, Hussain SZ, et al. Natural products/bioactive compounds as a source of anticancer drugs. Cancers (Basel) 2022;14(24):6203.##Tafrihi M, Toosi S, Minaei T, Gohari AR, Niknam V, Arab Najafi SM. Anticancer properties of Teucrium persicum in PC-3 prostate cancer cells. Asian Pac J Cancer Prev 2014;15(2):785-91.##Sadeghi Z, Yang J-L, Venditti A, Moridi Farimani M. A review of the phytochemistry, ethnopharmacology and biological activities of Teucrium genus (Germander). Nat Prod Res 2022;36(21):5647-64.##Sharifi-Rad M, Pohl P, Epifano F, Zengin G, Jaradat N, Messaoudi M. Teucrium polium (L): phytochemical screening and biological activities at different phenological stages. Molecules 2022;27( 5):1561.##Monsef-Esfahani H, Miri A, Amini M, Amanzadeh Y, Hadjiakhoondi A, Hajiaghaee R, et al. Seasonal variation in the chemical composition, antioxidant activity and total phenolic content of Teucrium persicum Boiss. essential oils. Res J Biol Sci 2010;5(7):492-8.##Rajabalian S. Methanolic extract of Teucrium polium L potentiates the cytotoxic and apoptotic effects of anticancer drugs of vincristine, vinblastine and doxorubicin against a panel of cancerous cell lines. Exp Oncol 2008;30(2):133-8.##Naeimi A, Tafrihi M, Mohadjerani M. Antioxidant and cytotoxic potentials of the methanolic extract of Teucrium persicum Boiss. in A-375 melanoma cells. Avicenna J Phytomed 2022;12(2):185-96.##Hajipour P, Eizadifard F, Tafrihi M. Chemical constituents, antioxidant and cytotoxic potential of chloroform and ethyl acetate extracts of Teucrium persicum. Jentashapir Journal of Cellular and Molecular Biology. 2022 Jun 30;13(2).##Tamokou DJD, Mpetga DJS, Lunga PK, Tene M, Tane P, Kuiate JR. Antioxidant and antimicrobial activities of ethyl acetate extract, fractions and compounds from stem bark of Albizia adianthifolia (Mimosoideae). BMC Complement Altern Med 2012; 12:99.##Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. cell. 2011; 144(5):646-74.##Rudzińska A, Juchaniuk P, Oberda J, Wiśniewska J, Wojdan W, Szklener K, et al. Phytochemicals in cancer treatment and cancer prevention—review on epidemiological data and clinical trials. Nutrients 2023;15(8):1896.##Key T. Fruit and vegetables and cancer risk. Br J Cancer. 2011;104(1):6-11.##Turati F, Rossi M, Pelucchi C, Levi F, La Vecchia C. Fruit and vegetables and cancer risk: a review of southern European studies. Br J Nutr 2015;113 Suppl 2:S102-S10. ##Surh YJ. Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer 2003;3(10):768-80.##Henchiri H, Bodo B, Deville A, Dubost L, Zourgui L, Raies A, et al. Sesquiterpenoids from Teucrium ramosissimum. Phytochemistry 2009;70(11-12):1435-41.##Wu ZK, Li HY, Zhu YL, Xiong MQ, Zhong JX. Neuroprotective and anti-inflammatory effects of eicosane on glutamate and NMDA-induced retinal ganglion cell injury. Int J Ophthalmol. 2024;17(4):638-45.##Chuah XQ, Okechukwu PN, Amini F, Teo SS. Eicosane, pentadecane and palmitic acid: The effects in: In vitro: Wound healing studies. Asian Pac J Trop Biomed 2018;8(10):490-9.##Sajeev Wagle S, Anne Lee J, Vasantha Rupasinghe HP. Synergistic Cytotoxicity of Extracts of Chaga Mushroom and Microalgae against Mammalian Cancer Cells In Vitro. Oxidative Medicine and Cellular Longevity January 2024;2024(12):1-13.##Miri A, Sharifi-Rad J, Tabrizian K, Nasiri AK. Antinociceptive and Anti-Inflammatory Activities of Teucrium persicum Boiss. Extract in Mice. 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Exploring the Role of hsa_circ_0052112 as a Potential Biomarker in Breast Cancer: Insights from Experimental and In Silico Analyses 2 2 Background: Circular RNAs (circRNAs) are important in tumorigenesis and cancer progression, highlighting their potential as biomarkers for diagnosis, prognosis, and treatment monitoring. Methods: This study consists of experimental and in silico phases. In the experimental phase, the expression of hsa_circ_0052112 in tumor and blood samples from 40 breast cancer women was analyzed, compared to the control group using Sybr Green real-time RT-PCR followed by total RNA extraction and cDNA synthesis. Statistical analysis was performed using the beta-actin gene as a normalizer, compared to the normal control group as a fold change. In the in silico phase, interactions among circRNA, RNA-Binding Proteins (RBPs), and microRNAs (miRNAs) were investigated using Interactome Database. The miRcancer database was utilized to assess breast cancer-related miRNAs linked to hsa_circ_0052112. Target miRNAs were identified with TargetScan and filtered for relevance through DisGeNET. K-means clustering grouped genes by expression patterns, visualized in Cytoscape to illustrate circRNA-miRNA-mRNA relationships. Hub genes underwent pathway enrichment analysis using Reactome database to determine their functional significance. Results: Data revealed a significant increase in hsa_circ_0052112 expression in both blood and tumour of breast cancer patients. This increase was especially pronounced in patients with estrogen, progesterone, and HER2 receptor positivity, as well as in advanced disease stages with lymph node involvement. Enrichment analysis of hub genes indicates their role in the PI3K/AKT signaling pathway. Conclusion: hsa_circ_0052112 shows promise as a multifaceted biomarker for breast cancer, enhancing diagnosis and prognosis; while supporting personalized treatment strategies. Further clinical validation is necessary to confirm its utility. 136 146 Mahdi Alizadeh Department of Medical Genetics, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) Department of Medical Genetics, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB) Iran Mahdieh Salimi Breast neoplasmsCircular RNAshsa_circ_0052112MicroRNAsPrognosis 60612.pdf Feinberg AP, Vogelstein B. Hypomethylation distinguishes genes of some human cancers from their normal counterparts. Nature 1983;301(5895):89-92.##Baylin SB, Jones PA. A decade of exploring the cancer epigenome - biological and translational implications. Nat Rev Cancer 2011;11(10):726-34.##Schröder R, Illert AL, Erbes T, Flotho C, Lübbert M, Duque-Afonso J. 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Plasma Circular-RNA 0005567 as a Potential Marker of Disease Activity in Rheumatoid Arthritis. Int J Mol Sci 2023;25(1):417.##Vellan CJ, Islam T, De Silva S, Mohd Taib NA, Prasanna G, Jayapalan JJ. Exploring novel protein-based biomarkers for advancing breast cancer diagnosis: A review. Clin Biochem 2024;129:110776.##Xu J, Gao H, Guan X, Meng J, Ding S, Long Q, et al. Circulating tumor DNA: from discovery to clinical application in breast cancer. Front Immunol 2024;15:1355887.##Zhu M, Gao Y, Zhu K, Yuan Y, Bai H, Meng L. Exosomal miRNA as biomarker in cancer diagnosis and prognosis: A review. Medicine (Baltimore) 2024;103(42):e40082.##Li X, Kazan H, Lipshitz HD, Morris QD. Finding the target sites of RNA-binding proteins. Wiley Interdiscip Rev RNA 2014;5(1):111-30.##Nakanishi K. Anatomy of four human Argonaute proteins. Nucleic Acids Res 2022; 50(12):6618-38.##Hu B, Chen R, Jiang M, Xiong S, Liu X, Fu B. 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Bridging the Skills Gap: Insights and Recommendations for Updating Medical Biotechnology Master's Curriculum in Iran 2 2 Background: Biotechnology is a rapidly developing field, and Iran aims to enhance its position in biosimilars through improved educational frameworks. Successful nations, such as the U.S., UK, Switzerland, China, and India, exemplify the positive impact of targeted educational programs in producing skilled graduates who advance the biotech sector. Research indicates a significant connection between curriculum relevance and graduate employability, suggesting that comprehensive curriculum assessment and reform can enhance student success and faculty involvement. This study investigated the necessity of revising the Master's curriculum in Medical Biotechnology in Iran to align with industry demands and global advancements. Methods: The current curriculum's effectiveness using the Delphi method was assessed to survey Master's students, professors, and industry stakeholders. Results: Findings revealed that while the current curriculum moderately aligns with educational objectives, significant gaps exist in practical training and resource availability. Many students reported feeling inadequately prepared for employment, particularly in essential skills such as animal cell culture, vaccine and monoclonal antibody design and production, and human skills like effective communication and teamwork. To address these deficiencies, some new courses focusing on practical experience and interdisciplinary approaches were recommended. Recommendations included enhancing laboratory facilities, integrating internships, and adopting team-based learning methods to improve student engagement and skill acquisition. Conclusion: Continuous investment in biotechnology education is crucial for maintaining competitive advantages in a globalized market. Overall, this research highlighted the importance of adapting educational programs to meet the dynamic needs of the biotechnology industry, ensuring graduates possess the necessary skills for successful careers in this field. 147 156 Amirhossein Ahmadieh-Yazdi Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Hamadan University of Medical Sciences Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Hamadan University of Medical Sciences Iran Behzad Imani Department of Operating Room, School of Paramedicine, Hamadan University of Medical Sciences Department of Operating Room, School of Paramedicine, Hamadan University of Medical Sciences Iran Akram Jalali Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Hamadan University of Medical SciencesResearch Center for Molecular Medicine, Institute of Cancer, Avicenna Health Research Institute, Hamadan University of Medical Sciences Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Hamadan University of Medical SciencesResearch Center for Molecular Medicine, Institute of Cancer, Avicenna Health Research Institute, Hamadan University of Medical Sciences IranIran Fahimeh Piryaei Research Center for Molecular Medicine, Institute of Cancer, Avicenna Health Research Institute, Hamadan University of Medical SciencesDepartment of Medical Genetics, School of Medicine, Hamadan University of Medical Sciences Research Center for Molecular Medicine, Institute of Cancer, Avicenna Health Research Institute, Hamadan University of Medical SciencesDepartment of Medical Genetics, School of Medicine, Hamadan University of Medical Sciences IranIran Razieh Dalirfardouei Research Center for Molecular Medicine, Institute of Cancer, Avicenna Health Research Institute, Hamadan University of Medical Sciences Research Center for Molecular Medicine, Institute of Cancer, Avicenna Health Research Institute, Hamadan University of Medical Sciences Iran CurriculumInternship and residencyIranMaster of scienceMedical biotechnology 60613.pdf Ezemba CC, Osuala OJ, Anakwenze VN. 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InScience and innovations in Iran: Development, progress, and challenges 2013 Jan 7 (pp. 139-157). New York: Palgrave Macmillan US.##Cardozo Ruíz Díaz E, Quintana SA, Rojas CM, Fernández Ríos D. Building bio-innovation systems through advanced biotechnology education. Front Bioeng Biotechnol 2024 Jul 9;12:1415103.##https://www.abdn.ac.uk/study/postgraduate-taught/degree-programmes/1036/msc-biotechnology-bioinformatics-and-bio-business.##https://www.rmit.edu.au/study-with-us/levels-of-study/postgraduate-study/masters-by-coursework/master-of-biotechnology-mc111.##https://landing.advanced.jhu.edu/msbiotech-aff/?utm_source=HML&utm_campaign=biotechnologydegrees&utm_content=list&utm_term=online&viq_channel=af.##https://www.westminster.ac.uk/biological-and-biomedical-sciences-courses/2025-26/september/full-time/applied-biotechnology-msc.##https://btp.stanford.edu/our-program/program-requirements.##Korenblit J. Biotechnology innovations in developing nations. Biotechnol Healthc 2006 Feb;3(1):55-8.##Chidobi RU, Menkiti BU. The Role of Educational Awareness of Biotechnology Product and Services in Senior Secondary Schools in Enugu East Local Government Area. International Journal of Environment, Agriculture and Biotechnology (IJEAB) 2017;2(3):1192-201.##Mohd Saruan N, Sagran A, Fadzil KS, Razali Z, Ow Phui San R, Somasundram C. Connecting learners: The role of biotechnology programme in preparing students for the industry. Biochem Mol Biol Educ 2015 Nov-Dec;43(6):460-7.##https://hcmep.behdasht.gov.ir/Arshad_Payeh.##Mahzari M, AlNahedh T, Ahmed AA, Al Rumyyan A, Shaban S, Magzoub ME. Practical Guide to Undergraduate Medical Curriculum Alignment and Mapping. Adv Med Educ Pract 2023 Sep 18;14:1001-12.##https://catalog.uic.edu/gcat/course-descriptions/mbt/.##https://www.fh-krems.ac.at/en/study/master/full-time/medical-and-pharmaceutical-biotechnology/.##https://www.napier.ac.uk/courses/msc-medical-biotechnology-postgraduate-fulltime.##Sahebihag MH, Khadivi AA, Soheili A, Moghbeli G, Khaje Goudari M, Valizadeh L. The challenges of nursing doctoral curriculum in Iran: a critical look based on Delphi technique. Nursing And Midwifery Journal 2017 Sep 10;15(6):424-39.##Mohammadzade A, Mortazavi F, Safavinaini S, Tabatabi S. Stakeholders, Comments about the Need Assessment for the Development and Implementation of Curriculum In Hearing-Speech and Language Sciences at PhD Level. Medical Education Journal 2013;1(2):27-36.##Nouroozzadeh R, Mahmoodi R, Fathi Vajargah K, Nave Ebrahim AR. The Universities Participation Status in Revising the Curricula Approved by the Higher Council for Planning. 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Toxicity Study of Silver Nanoparticles Synthesized from Suaeda Monoica on Hep-2 Cell Line 2 2 This is to inform that the authors mentioned below have requested a correction in the published paper as below: Satyavani K, Gurudeeban S, Ramanathan T, Balasubramanian T. "Toxicity Study of Silver Nanoparticles Synthesized from Suaeda Monoica on Hep-2 Cell Line" Avicenna Journal of Medical Biotechnology. 2012 Jan;4(1):35. Revised Figure Legend: Figure 1. Representative AFM image of nanoparticles synthesized from different plant species (Citrullus colocynthis and Suaeda monoica) using callus and leaf extracts. Despite originating from distinct plant sources, the nanoparticles exhibit uniform morphology and an average size of ~31 nm. The consistency in nanoparticle size and shape supports the reproducibility of the synthesis process. 157 157 Editorial 60614.pdf ####