Monoclonal Antibodies and the Rise of Precision Immunopsychiatry 2 2 No Abstract 94 95 Shahin Akhondzadeh Editorial 70644.pdf Abbasian F, Bagheri S, Moradi K, Keykhaei M, Etemadi A, Shalbafan M, et al. Evidence for Anti-inflammatory Effects of Adalimumab in Treatment of Patients with Major Depressive Disorder: A Pilot, Randomized, Controlled Trial. Clin Neuropharmacol 2022;45(5):128-134.##Motamed M, Karimi H, Sanjari Moghaddam H, Taherzadeh Boroujeni S, Sanatian Z, Hasanzadeh A, et al. Risperidone combination therapy with adalimumab for treatment of chronic schizophrenia: a randomized, double-blind, placebo-controlled clinical trial. Int Clin Psychopharmacol 2022;37(3):92-101. ##Abbasi SH, Behpournia H, Ghoreshi A, Salehi B, Raznahan M, Rezazadeh SA, et al. The effect of mirtazapine add on therapy to risperidone in the treatment of schizophrenia: a double-blind randomized placebo-controlled trial. Schizophr Res 2010;116(2-3):101-6.##Akhondzadeh S, Milajerdi MR, Amini H, Tehrani-Doost M. Allopurinol as an adjunct to lithium and haloperidol for treatment of patients with acute mania: a double-blind, randomized, placebo-controlled trial. Bipolar Disord 2006;8(5 Pt 1):485-9.##Ghaleiha A, Ghyasvand M, Mohammadi MR, Farokhnia M, Yadegari N, Tabrizi M, et al. Galantamine efficacy and tolerability as an augmentative therapy in autistic children: A randomized, double-blind, placebo-controlled trial. J Psychopharmacol 2014;28(7):677-85.##Jafari P, Ghanizadeh A, Akhondzadeh S, Mohammadi MR. Health-related quality of life of Iranian children with attention deficit/hyperactivity disorder. Qual Life Res 2011; (1):31-6.##Kouti L, Noroozian M, Akhondzadeh S, Abdollahi M, Javadi MR, Faramarzi MA, Mousavi S, et al. Nitric oxide and peroxynitrite serum levels in Parkinson's disease: correlation of oxidative stress and the severity of the disease. Eur Rev Med Pharmacol Sci 2013;17(7):964-70.##

Luteolin-Treated 4T1 Cell-Derived Exosomes as Novel Antiproliferative Agents In Vitro and In Vivo 2 2 Background: Breast cancer is the most widespread malignancy among women worldwide. Luteolin, a flavonoid, has demonstrated anti-cancer effects by triggering apoptosis in tumor cells. Exosomes are gaining much attention for cancer therapeutic approaches due to multitude of beneficial effects. This study is aimed to investigate the possible potential of exosomes derived from luteolin-treated 4T1 cells to ameliorate tumor in comparison to luteolin treatment only. Methods: In this study, 4T1 cell culture was exposed to luteolin. Following exosome extraction, they were characterized using field emission scanning electron microscopy, dynamic light scattering and western blot analysis. MTT assay was performed in order to evaluate cell viability after exposure to different concentrations of luteolin and exosomes. An in vivo breast cancer model was induced via subcutaneous injection of 4T1 cells to the BALB/C mice. After 14 days, tumor volume was measured, and expression of RhoA and ERK mRNAs were quantified by Real Time PCR. Results: The MTT assay demonstrated that exosomes from luteolin-treated 4T1 cells at a concentration of 320 μg/μl reduced cell viability by approximately 70% in a dose-dependent manner. Tumor volume in the exosome-treated group decreased by 57% relative to the tumor group, while the luteolin-treated group demonstrated a 39% reduction. Furthermore, RhoA gene expression was substantially downregulated in the exosome-treated group, and exosomes were more effective than luteolin in reducing ERK gene expression. Conclusion: Exosomes derived from luteolin-treated 4T1 cells effectively suppress breast cancer cell growth by reducing 4T1 cell viability and by decreasing tumor volume and downregulating tumor-associated genes RhoA and ERK. These results propose a novel therapeutic strategy for breast cancer, highlighting the promising potential of exosomes as an efficient drug delivery system. 96 103 Pardis Heidari Faculty of Veterinary Sciences, SR.C., Islamic Azad University Faculty of Veterinary Sciences, SR.C., Islamic Azad University Iran Razieh Hosseini Nika Nasihatgar Faculty of Veterinary Sciences, SR.C., Islamic Azad University Faculty of Veterinary Sciences, SR.C., Islamic Azad University Iran Breast neoplasmsCell survivalExosomesLuteolinTumor burden 70645.pdf Xu Y, Gong M, Wang Y, Yang Y, Liu S, Zeng Q. Global trends and forecasts of breast cancer incidence and deaths. Sci Data 2023;10(1):334.##Rauf A, Imran M, Butt MS, Nadeem M, Peters DG, Mubarak MSJCrifs, et al. Resveratrol as an anti-cancer agent: A review. Crit Rev Food Sci Nutr 2018;58(9):1428-47.##Zhang QY, Wang FX, Jia KK, Kong LD. Natural product interventions for chemotherapy and radiotherapy-induced side effects. Front Pharmacol 2018;9:1253.##Seyed MA, Jantan I, Bukhari SNA, Vijayaraghavan KJJoa. A comprehensive review on the chemotherapeutic potential of piceatannol for cancer treatment, with mechanistic insights. J Agric Food Chem 2016;64(4):725-37.##Zhang HW, Hu JJ, Fu RQ, Liu X, Zhang YH, Li J, et al. Flavonoids inhibit cell proliferation and induce apoptosis and autophagy through downregulation of PI3Kγ mediated PI3K/AKT/mTOR/p70S6K/ULK signaling pathway in human breast cancer cells. Sci Rep 2018;8(1):11255.##Yan W, Ma X, Zhao X, Zhang S. Baicalein induces apoptosis and autophagy of breast cancer cells via inhibiting PI3K/AKT pathway in vivo and vitro. Drug design, development and therapy. Drug Des Devel Ther 2018;12:3961-72.##Chen Z, Kong S, Song F, Li L, Jiang H. Pharmacokinetic study of luteolin, apigenin, chrysoeriol and diosmetin after oral administration of Flos Chrysanthemi extract in rats. Fitoterapia 2012;83(8):1616-22.##Krifa M, Leloup L, Ghedira K, Mousli M, Chekir-Ghedira L. Luteolin induces apoptosis in BE colorectal cancer cells by downregulating calpain, UHRF1, and DNMT1 expressions. Nut Cancer 2014;66(7):1220-7.##Chen Z, Zhang B, Gao F, Shi R. Modulation of G2/M cell cycle arrest and apoptosis by luteolin in human colon cancer cells and xenografts. Oncol Lett 2018;15(2):1559-65.##Lin HW, Shen TJ, Yang NC, Wang M, Hsieh WC, Chuang CJ, et al. Luteolin reduces aqueous extract PM2. 5-induced metastatic activity in H460 lung cancer cells. Int J Med Sci 2022;19(10):1502-9.##Johnsen KB, Gudbergsson JM, Skov MN, Pilgaard L, Moos T, Duroux M. A comprehensive overview of exosomes as drug delivery vehicles—endogenous nanocarriers for targeted cancer therapy. Biochim Biophys Acta 2014;1846(1):75-87.##Nam GH, Choi Y, Kim GB, Kim S, Kim SA, Kim ISJAM. Emerging prospects of exosomes for cancer treatment: from conventional therapy to immunotherapy. Adv Mater 2020;32(51):e2002440.##Kim MS, Haney MJ, Zhao Y, Mahajan V, Deygen I, Klyachko NL, et al. Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells. Nanomedicine 2016;12(3):655-64.##Ji R, Zhang X, Gu H, Ma J, Wen X, Zhou J, et al. miR-374a-5p: a new target for diagnosis and drug resistance therapy in gastric cancer. Mol Ther Nucleic Acids 2019;18:320-31.##Kim H, Kim EH, Kwak G, Chi SG, Kim SH, Yang Y. Exosomes: cell-derived nanoplatforms for the delivery of cancer therapeutics. Int J Mol Sci 2020;22(1):14.##Wang J, Zheng Y, Zhao M. Exosome-based cancer therapy: implication for targeting cancer stem cells. Front Pharmacol 2017;7:533.##Rengarajan T, Nandakumar N, Rajendran P, Haribabu L, Nishigaki I, Balasubramanian MPJAPJoCP. D-pinitol promotes apoptosis in MCF-7 cells via induction of p53 and Bax and inhibition of Bcl-2 and NF-κB. Asian Pac J Cancer Prev 2014;15(4):1757-62.##Montaser R, Luesch HJFmc. Marine natural products: a new wave of drugs? Future Med Chem 2011;3(12):1475-89.##Devi KP, Rajavel T, Nabavi SF, Setzer WN, Ahmadi A, Mansouri K, et al. Hesperidin: A promising anticancer agent from nature. Industrial Crops and Products 2015 Dec 15;76:582-9.##Lin Y, Shi R, Wang X, Shen H-MJCcdt. Luteolin, a flavonoid with potential for cancer prevention and therapy. Curr Cancer Drug Targets 2008;8(7):634-46.##Wruck C, Claussen M, Fuhrmann G, Römer L, Schulz A, Pufe T, et al. Luteolin protects rat PC 12 and C6 cells against MPP+ induced toxicity via an ERK dependent Keapl-Nrf2-ARE pathway. Neuropsychiatric disorders an integrative approach: Springer; 2007. p. 57-67.##Birt DF, Hendrich S, Wang WJP. Dietary agents in cancer prevention: flavonoids and isoflavonoids. Pharmacol Ther 2001;90(2-3):157-77.##Imran M, Rauf A, Abu-Izneid T, Nadeem M, Shariati MA, Khan IA, et al. Luteolin, a flavonoid, as an anticancer agent: A review. Biomed Pharmacother 2019;112:108612.##Qu Z, Wu J, Wu J, Luo D, Jiang C, Ding Y. Exosomes derived from HCC cells induce sorafenib resistance in hepatocellular carcinoma both in vivo and in vitro. J Exp Clin Cancer Res 2016;35(1):159.##Zhu L, Kalimuthu S, Gangadaran P, Oh JM, Lee HW, Baek SH, et al. Exosomes derived from natural killer cells exert therapeutic effect in melanoma. Theranostics 2017;7(10):2732-45.##Jang SC, Kim OY, Yoon CM, Choi DS, Roh TY, Park J, et al. Bioinspired exosome-mimetic nanovesicles for targeted delivery of chemotherapeutics to malignant tumors. ACS Nano 2013;7(9):7698-710.##Melzer C, Rehn V, Yang Y, Bähre H, von der Ohe J, Hass R. Taxol-loaded MSC-derived exosomes provide a therapeutic vehicle to target metastatic breast cancer and other carcinoma cells. Cancers (Basel) 2019;11(6):798.##Peak TC, Praharaj PP, Panigrahi GK, Doyle M, Su Y, Schlaepfer IR, et al. Exosomes secreted by placental stem cells selectively inhibit growth of aggressive prostate cancer cells. Biochem Biophys Res Commun 2018;499(4):1004-10.##Kheradjoo H, Nouralishahi A, Hoseinzade Firozabdi MS, Mohammadzadehsaliani S. Mesenchymal stem/stromal (MSCs)-derived exosome inhibits retinoblastoma Y-79 cell line proliferation and induces their apoptosis. Nanomedicine Research Journal 2022;7(3):264-9.##Haga RB, Ridley AJ. Rho GTPases: Regulation and roles in cancer cell biology. Small GTPases 2016;7(4):207-21.##Kim JG, Islam R, Cho JY, Jeong H, Cap KC, Park Y, et al. Regulation of RhoA GTPase and various transcription factors in the RhoA pathway. J Cell Physiol 2018;233(9):6381-92.##Zhang S, Tang Q, Xu F, Xue Y, Zhen Z, Deng Y, et al. RhoA regulates G1-S progression of gastric cancer cells by modulation of multiple INK4 family tumor suppressors. Mol Cancer Res 2009;7(4):570-80.##Kohno M, Pouyssegur J. Targeting the ERK signaling pathway in cancer therapy. Annals of medicine. 2006;38(3):200-11.##Sugiura R, Satoh R, Takasaki T. ERK: a double-edged sword in cancer. ERK-dependent apoptosis as a potential therapeutic strategy for cancer. Cells 2021;10(10):2509.##Maik-Rachline G, Hacohen-Lev-Ran A, Seger R. 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Investigation of the Molecular Signature of Senescence in Mesenchymal Stem Cells 2 2 Background: Mesenchymal Stem Cells (MSCs) play a pivotal role in regenerative medicine due to their multipotency and immunomodulatory properties. However, during in vitro expansion, MSCs undergo senescence, characterized by a decline in proliferation, impairment of differentiation potential, and altered secretory profiles, which limits their therapeutic efficacy. This study aimed to identify novel molecular regulators and net-work-level interactions underlying MSC senescence through microarray analysis of the GSE7888 dataset, comparing early and senescent MSCs. Methods: A total of 4597 Differentially Expressed Genes (DEGs) were identified be-tween early (passages 4-5) and senescent (passages 22-28) MSCs, with 2219 upregulated and 2379 downregulated. Key regulators such as CDKN1A (p21), CDK4, and CDK6 were implicated in cell cycle arrest and the progression of senescence. Pathway analysis highlighted the mTOR, FoxO, and p53 signaling pathways as key regulators of stress responses, metabolism, and aging. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis highlighted cellular processes, including protein transport and intracellular signaling, while protein-protein interaction networks identified high-connectivity nodes such as TP53, FOXO3, and MDM2. Senescent MSCs displayed phenotypic changes, including altered morphology and the emergence of the Senescence-Associated Secretory Phenotype (SASP), which impaired re-generative potential. Results: The findings suggest that targeting the mTOR, FoxO, and p53 pathways could delay senescence and enhance MSC therapeutic potential. Interventions such as rapamycin and FoxO3 activators show promise in reversing senescence. Conclusion: Future research should explore small molecules and gene-based therapies targeting senescence pathways to improve MSC-based regenerative strategies. These results provide a foundation for developing innovative approaches to optimize MSC ap-plications in clinical therapies. 104 112 Forough-Azam Sayahpour Medical College of Wisconsin, Center of Cancer Discovery, Milwaukee Medical College of Wisconsin, Center of Cancer Discovery, Milwaukee United States of America Marjan Nejati Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biolo-gy and Technology, ACECRDepartment of Cell and Molecular Biology, School of Biology, College of Sciences, University of Tehran Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biolo-gy and Technology, ACECRDepartment of Cell and Molecular Biology, School of Biology, College of Sciences, University of Tehran IranIran Mahya Rouhollahi-Masoumi Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biolo-gy and Technology, ACECR Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biolo-gy and Technology, ACECR Iran Mohammadreza Baghaban Eslaminejad Sara Taleahmad Gene ontologyMesenchymal stem cellsRegenerative medicineSenescence-associated secretory phenotype 70646.pdf Li Y, Wu Q, Wang Y, Li L, Bu H, Bao J. Senescence of mesenchymal stem cells. Int J Mol Med 2017;39(4):775-82.##Trounson A, McDonald C. Stem cell therapies in clinical trials: progress and challenges. Cell Stem Cell 2015;17(1):11-22.##Han Y, Li X, Zhang Y, Han Y, Chang F, Ding J. Mesenchymal stem cells for regenerative medicine. Cells 2019;8(8):886.##Lan T, Luo M, Wei X. Mesenchymal stem/stromal cells in cancer therapy. Journal of Hematology & Oncology. 2021;14:1-16.##Kim GB, Shon OJ, Seo MS, Choi Y, Park WT, Lee GW. Mesenchymal stem cell-derived exosomes and their therapeutic potential for osteoarthritis. Biology (Basel) 2021;10(4):285.##Chen H, Liu O, Chen S, Zhou Y. Aging and mesenchymal stem cells: therapeutic opportunities and challenges in the older group. Gerontology 2022;68(3):339-52.##Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell 2011;144(5):646-74.##Campisi J. Aging, cellular senescence, and cancer. 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Chitosan hydrogel-loaded MSC-derived extracellular vesicles promote skin rejuvenation by ameliorating the senescence of dermal fibroblasts. Stem Cell Res Ther 2021 Mar 20;12(1):196.##Sills AM, Artavia JM, DeRosa BD, Ross CN, Salmon AB. Long‐term treatment with the mTOR inhibitor rapamycin has minor effect on clinical laboratory markers in middle‐aged marmosets. Am J Primatol 2019;81(2):e22927.##Chang ZS, Xia JB, Wu HY, Peng WT, Jiang FQ, Li J, et al. Forkhead box O3 protects the heart against paraquat‐induced aging‐associated phenotypes by upregulating the expression of antioxidant enzymes. Aging Cell 2019;18(5):e12990.##Wu J, Huang S, Yu Y, Lian Q, Liu Y, Dai W, et al. Human adipose and synovial-derived MSCs synergistically attenuate osteoarthritis by promoting chondrocyte autophagy through FoxO1 signaling. Stem Cell Res Ther 2024 Aug 15;15(1):261.##

Potential Targets in Innate Immunity Receptors for Gastric Cancer: Insights from Virtual Screening in TCM and In Vitro Assay 2 2 Background: Gastric Cancer (GC) poses a substantial global health threat, ranking as the second leading cause of cancer-related mortality among gastrointestinal malignancies. This investigation explores the potential therapeutic implications of plant extracts on gastric cancer, with a specific focus on their effects on the innate immune system. Methods: A comprehensive analysis was conducted using 200 Sequence Read Runs (SRRs) thigh samples associated with gastrointestinal cancer tissue, juxtaposed with pathologically confirmed healthy tissues serving as controls. Differential Gene Expression (DGE) testing, encompassing the examination of 28,000 genes, including 95 pivotal genes associated with the innate immune system, was conducted. Findings elucidate alterations in the expression of key pattern recognition receptors, such as TLR2 and TLR4, as well as pivotal molecules within their signaling pathways. In pursuit of potential antagonists for these receptors, virtual screening on the Maestro docking platform in the Schrödinger 2022 package was conducted, evaluating 220,000 diverse tautomer’s of plant active substances. Selected candidates, exhibiting superior docking scores across four additional platforms, were subjected to further scrutiny. Results: MMT results showed that nettle extract showed significant cytotoxic effects within 12 hr compared to the control (no treatment) sample, resulting in a 34.7% reduction in AGS cancer cell viability. The flow cytometry test showed that the control group had 71%, and groups treated with nettle extract for two and 12 hr had 65.3 and 67.18% viable cells, respectively. These differences were not statistically significant, indicating that nettle extract selectively preserves healthy living cells. Conclusion: Cytotoxicity tests and cell cycle assessments confirmed the ability of nettle extract to reduce the survival of GC cells. This property makes nettle a promising candidate for drug development in this direction. 113 120 Abbas Ganjali University of Zabol University of Zabol Iran Baratali Fakheri Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Zabol Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Zabol Iran Abbas Bahari Leila Fahmideh Department of Plant Breeding and Biotechnology, Gorgan University of Agriculture Sciences and Natural Resources Department of Plant Breeding and Biotechnology, Gorgan University of Agriculture Sciences and Natural Resources Iran Reza Valadan Department of Immunology, Molecular and Cell Biology Research Centre, Faculty of Medicine, Mazandaran University of Medical Sciences Department of Immunology, Molecular and Cell Biology Research Centre, Faculty of Medicine, Mazandaran University of Medical Sciences Iran Mehdi Tavakolizadeh Department of Pharmacognosy, School of Pharmacy, Zanjan University of Medical Sciences Department of Pharmacognosy, School of Pharmacy, Zanjan University of Medical Sciences Iran Cell cycleDrug developmentGene expressionImmune systemPlant extractsSignal transductionToll-Like Receptor 2Toll-Like Receptor 4 70647.pdf Kiri S, Ryba T. 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Therapeutic Potential of Alternating Magnetic Fields for Normalizing Blood Parameters and Restoring Renal, and Cardiac Function in Diabetic Mice 2 2 Background: In recent years, the number of adults aged 20-79 years living with diabetes has increased more than threefold. Currently, the treatment of diabetes typically involves the long-term use of chemical and herbal drugs. However, prolonged use of chemical drugs may lead to side effects that can be detrimental to health. Therefore, this study aims to normalize blood glucose levels and restore kidney and heart cells. Methods: The research was conducted using diabetic mice as experimental subjects. The treatment involved exposure to an alternating Magnetic Field with Magnetic Flux Densities of 0.3 and 0.6 mT for 20 min/day over five consecutive days. The frequencies of the applied Magnetic Fields were 50, 100, 150, and 200 Hz. Results: The results showed that the greatest reduction in blood glucose levels (92.11%) was observed at a frequency of 100 Hz and a Magnetic Flux Density of 0.6 mT. Meanwhile, the highest increase in hemoglobin levels (81.11%) occurred at a frequency of 150 Hz and a Magnetic Flux Density of 0.3 mT. Other parameters that experienced non-linear changes included cholesterol levels, blood viscosity, and erythrocytes count, glomerulus and kidney cell density, and heart cell density. Conclusion: The optimal effects of magnetic field exposure do not always occur at the same frequency or Magnetic Flux Density. 121 130 Mokhamad Tirono Farid Samsu Hananto Department of Physics, Faculty of Science and Technology, State Islamic University of Maulana Malik Ibrahim of Malang Department of Physics, Faculty of Science and Technology, State Islamic University of Maulana Malik Ibrahim of Malang Indonesia BloodBlood glucoseDiabetes mellitusHeartKidney 70648.pdf Fuentes E, Fuentes M, Alarcón M, Palomo I. Immune System Dysfunction in the Elderly. An Acad Bras Cienc 2017 Jan-Mar;89(1):285-299.##Aiyar S, Ebeke C, Shao X. The Impact of Workforce Aging on European. Int Monet Fund 2016;16:1-28.##Federation ID. IDF Diabetes Atlas 2021. International Diabetes Federation. 2021.##Galicia-Garcia U, Benito-Vicente A, Jebari S, Larrea-Sebal A, Siddiqi H, Uribe KB, Ostolaza H, Martín C. Pathophysiology of Type 2 Diabetes Mellitus. 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Evaluation of the Anti-melanogenic Effect of Aqueous and Hydroalcoholic Extracts of Nasturtium officinale on the B16F10 Cell Line 2 2 Background: This project aimed to evaluate the anti-melanogenic characteristics of Nasturtium officinale (N. officinale) by assessing the impact of both aqueous and hydroalcoholic extracts on the inhibition of cellular and mushroom tyrosinase enzymes, as well as the suppression of the melanin synthesis in B16F10 melanoma cells. Methods: The aerial components of N. officinale were subjected to extraction using distilled water: ethanol (7:3) through the maceration technique. The extract’s phenolic compounds were quantified employing the Folin-Ciocalteu method. The evaluation of the safety profile of the extracts on B16F10 cells was done by the MTT assay. Subsequently, the melanin concentration in B16F10 cells, alongside the inhibitory effects on both mushroom and cellular tyrosinase, was assessed following treatment with the aforementioned extracts. Results: The aqueous and hydroalcoholic extracts exhibited no significant toxicity on B16F10 when compared to Phosphate-Buffered Saline (PBS). Additionally, there was no notable difference in the cytotoxic effects of extracts on the B16F10 cell line. Both extracts resulted in inhibition of cellular and mushroom tyrosinase, along with a decrease in melanin levels in B16F10 in a concentration-dependent manner. Ultimately, the total phenolic content in the aqueous and hydroalcoholic extracts was found to be approximately 14 and 30 mg/g of gallic acid, respectively. Conclusion: This in vitro investigation offers evidence supporting the skin brightening properties of N. officinale as an anti-melanogenic agent. Given its safety profile and absence of toxic effects on melanoma cells, it may be incorporated into the formulation of skin-brightening products following preclinical tests. 131 136 Mohaddese Heydari Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences Iran Masoud Sadeghi-Dinani Department of Pharmacognosy, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences Department of Pharmacognosy, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences Iran Fatemeh Shafiee MelaninMelanoma cell lineNasturtium officinaleTyrosinase 70649.pdf Zhou C, Lee C, Salas J, Luke J. Guide to tinted sunscreens in skin of color. 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Agar and Agitated Microshoot Culture Media on Glucosinolate and Phenolic Acid Production, and Antioxidant Activity. Biomolecules 2020 Aug 21;10(9):1216. ##Boligon AA, Janovik V, Boligon AA, Pivetta CR, Pereira RP, Rocha JBT Da, et al. HPLC analysis of polyphenolic compounds and antioxidant activity in nasturtium officinale. Int J Food Prop 2013;16(1):61–9.##Panahi Kokhdan E, Khodabandehloo H, Ghahremani H, Doustimotlagh AH. A Narrative Review on Therapeutic Potentials of Watercress in Human Disorders. Evid Based Complement Alternat Med 2021 May 7:2021:5516450. ##Sedaghattalab M, Razazan M, Sadeghi H, Doustimotlagh AH, Toori MA, Abbasi Larki R, et al. Effects of Nasturtium officinale Extract on Antioxidant and Biochemical Parameters in Hemodialysis Patients: A Randomized Double‐Blind Clinical Trial. Evid Based Complement Alternat Med 2021;2021(1):1632957. ##Giallourou N, Oruna-Concha MJ, Harbourne N. Effects of domestic processing methods on the phytochemical content of watercress (Nasturtium officinale). Food Chem 2016 Dec 1;212:411-9.##Klimek-Szczykutowicz M, Szopa A, Dziurka M, Komsta Ł, Tomczyk M, Ekiert H. The influence of Nasturtium officinale R. Br. Agar and agitated microshoot culture media on glucosinolate and phenolic acid production, and antioxidant activity. Biomolecules 2020;10(9):1216. ##Al-Snafi AE. A review on Nasturtium officinale: A potential medicinal plant. IOSR Journal of Pharmacy 2020;10(9):33-43.##Yazdanparast R, Bahramikia S, Ardestani A. Nasturtium officinale reduces oxidative stress and enhances antioxidant capacity in hypercholesterolaemic rats. Chem Biol Interact 2008 Apr 15;172(3):176-84. ##Alaghebandi S, Yazdiniapour Z, Shafiee F. In Vitro Evaluation the Anti-Melanogenic Effects of Various Extracts of Oat Seeds: Anti-Melanogenic Effects of Oat Seeds. Iranian Journal of Pharmaceutical Sciences 2022;18(2):95–104.##Baurin N, Arnoult E, Scior T, Do PB QT. Preliminary screening of some tropical plants for anti-tyrosinase activity. J Ethnopharmacol 2002;82(2-3):155-8. ##Nilash AB, Jahanbani J, Jolehar M. Effect of Nasturtium Extract on Oral Cancer. Adv Biomed Res 2023;12(25):1–6. ##Casanova NA, Ariagno JI, López Nigro MM, Mendeluk GR, Gette M de los A, Petenatti E, et al. In vivo antigenotoxic activity of watercress juice (nasturtium officinale) against induced DNA damage. J Appl Toxicol 2013;33(9):880–5. ##Chang Lee-Wen LJJ, Wang BS, Wang MY, Tai HM, Hung WJ, Chen YJ, et al. Antioxidant and antityrosinase activity of mulberry (Morus alba L.) twigs and root bark. Food Chem Toxicol 2011 Apr;49(4):785-90. ##Wang JJ, Wu CC, Lee CL, Hsieh SL, Chen JB, Lee CI. Antimelanogenic, antioxidant and antiproliferative effects of Antrodia camphorata fruiting bodies on B16-F0 melanoma cells. PLoS One 2017;12(1):1–15. ##Hwang JH, Lee BM. Inhibitory effects of plant extracts on tyrosinase, L-DOPA oxidation, and melanin synthesis. 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Establishing an Optimized Caco-2/THP-1 Co-Culture Model to Efficiently Simulate Inflammatory Bowel Disease In Vitro 2 2 Background: Inflammatory Bowel Disease (IBD) is a complex disorder for which the mechanisms and targeted therapies remain unclear. Several in vitro models, including organoids, cytokine-stimulated Caco-2 monolayers, and co-culture systems, have been developed to study IBD pathogenesis and potential treatments. Meanwhile, the Caco-2/THP-1 co-culture is a practical model representing the interaction of intestinal epithelial and immune cells. However, multiple factors, such as culture duration and exposure time to inflammatory agents, significantly affect model outcomes. Developing an optimized co-culture that better mimics intestinal inflammation can introduce a valuable method for future studies. This study aimed to optimize a Caco-2/THP-1 co-culture model, focusing on culture timing and treatment conditions. Methods: THP-1 monocytes were differentiated into macrophage-like cells (M0) with phorbol 12-myristate 13-acetate (PMA, 50 ng/ml, 48 hr). M0 cells were treated with different Lipopolysaccharide (LPS) concentrations for 6 or 24 hr to determine the optimal inflammatory dose. Inflammatory macrophages (M1) were co-cultured with differentiated or undifferentiated Caco-2 monolayers. Expression of IL-6, IL-8, and TNF-α was measured by qRT-PCR, M1 macrophage markers (CD86/HLA-DR) by flow cytometry, and nitric oxide by the Griess assay. Results: Stimulation with 100 ng/ml LPS for 6 hr increased M1 (CD86⁺/HLA-DR⁺) macrophages to 58.9% and induced maximal nitric oxide production (179.3 µM). Co-culture with these M1 cells enhanced IL-8 and modestly increased IL-6 expression in differentiated Caco-2 cells compared with other groups. Conclusion: The differentiated Caco-2/THP-1 co-culture efficiently mimics intestinal inflammation observed in IBD and provides an optimized in vitro model for further investigations. 137 146 Gilda Parsamanesh Department of Immunology, School of Medicine, Tehran University of Medical SciencesGastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences Department of Immunology, School of Medicine, Tehran University of Medical SciencesGastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences IranIran Kaveh Baghaei Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical SciencesOlivia Newton-John cancer Research Institute and School of cancer Medicine, La Trobe University Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical SciencesOlivia Newton-John cancer Research Institute and School of cancer Medicine, La Trobe University IranAustralia Nima Rezaei Research Center for Immunodeficiencies, Children’s Medical Center Hospital, Tehran University of Medical SciencesNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN) Research Center for Immunodeficiencies, Children’s Medical Center Hospital, Tehran University of Medical SciencesNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN) IranIran Biological modelsCaco-2 cellsInflammatory bowel diseaseLipopeptidesMacrophage activation 70650.pdf Macedo MH, Dias Neto M, Pastrana L, Gonçalves C, Xavier M. Recent advances in cell‐based in vitro models to recreate human intestinal inflammation. Adv Sci (Weinh) 2023;10(31):2301391. ##Kaplan GG, Windsor JW. The four epidemiological stages in the global evolution of inflammatory bowel disease. Nat Rev Gastroenterol Hepatol 2021;18(1):56–66.##Kostic AD, Xavier RJ, Gevers D. The microbiome in inflammatory bowel disease: current status and the future ahead. Gastroenterology 2014;146(6):1489–99.##Tao Y, Li Y, Zhou T, Zhao L, Li P, Sun M, et al. Etiological mechanisms underlying the hazard factors and inflammatory bowel disease: a prospective cohort study. Sci Rep 2025;15(1):23517.##Chelakkot C, Ghim J, Ryu SH. Mechanisms regulating intestinal barrier integrity and its pathological implications. Exp Mol Med 2018;50(8):1–9.##Silaghi A, Constantin VD, Socea B, Banu P, Sandu V, Andronache LF, et al. Inflammatory bowel disease: pathogenesis, diagnosis and current therapeutic approach. J Mind Med Sci 2022;9(1):56–77.##Wang M, Shi J, Yu C, Zhang X, Xu G, Xu Z, et al. Emerging strategy towards mucosal healing in inflammatory bowel disease: what the future holds? Front Immunol 2023;14:1298186.##Guan Q, Zhang J. Recent advances: the imbalance of cytokines in the pathogenesis of inflammatory bowel disease. Mediators Inflamm 2017;2017(1):4810258.##Joshi A, Soni A, Acharya S. In vitro models and ex vivo systems used in inflammatory bowel disease. In Vitro Model 2022;1(3):213–27.##Lee CH, Koh S-J, Radi ZA, Habtezion A. Animal models of inflammatory bowel disease: novel experiments for revealing pathogenesis of colitis, fibrosis, and colitis-associated colon cancer. Intest Res 2023;21(3):295–305.##Wen C, Chen D, Zhong R, Peng X. Animal models of inflammatory bowel disease: category and evaluation indexes. Gastroenterol Rep (Oxf) 2024;12:goae021.##Oh SY, Cho K-A, Kang JL, Kim KH, Woo S-Y. Comparison of experimental mouse models of inflammatory bowel disease. 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A human Caco-2-based co-culture model of the inflamed intestinal mucosa for particle toxicity studies. In Vitro Model 2023;2(1-2):43–64.##Ferreira B, Barros AS, Leite-Pereira C, Viegas J, das Neves J, Nunes R, et al. Trends in 3D models of inflammatory bowel disease. Biochim Biophys Acta (BBA)-Molecular Basis Dis 2024;1870(3):167042.##Hobbs S, Reynoso M, Geddis A V, Mitrophanov AY, Matheny Jr RW. LPS‐stimulated NF‐κB p65 dynamic response marks the initiation of TNF expression and transition to IL‐10 expression in RAW 264.7 macrophages. Physiol Rep 2018;6(21):e13914. ##Unuvar Purcu D, Korkmaz A, Gunalp S, Helvaci DG, Erdal Y, Dogan Y, et al. Effect of stimulation time on the expression of human macrophage polarization markers. PLoS One 2022;17(3):e0265196.##Marescotti D, Lo Sasso G, Guerrera D, Renggli K, Ruiz Castro PA, Piault R, et al. Development of an advanced multicellular intestinal model for assessing immunomodulatory properties of anti-inflammatory compounds. 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Differential transcript and soluble factor patterns in macrophage/enterocyte-like monolayer co-cultures based on apical or basolateral LPS exposure. Front Immunol 2025;16:1527007. ##Sonnier DI, Bailey SR, Schuster RM, Lentsch AB, Pritts TA. TNF-α induces vectorial secretion of IL-8 in Caco-2 cells. J Gastrointest Surg 2010;14(10):1592–9.##Van De Walle J, Hendrickx A, Romier B, Larondelle Y, Schneider YJ. Inflammatory parameters in Caco-2 cells: Effect of stimuli nature, concentration, combination and cell differentiation. Toxicol Vitro 2010;24(5):1441–9.##Ferreira B, Ferreira C, Martins C, Nunes R, das Neves J, Leite-Pereira C, et al. Establishment of a 3D multi-layered in vitro model of inflammatory bowel disease. J Control Release 2025;377:675–88.##Haddad MJ, Zuluaga-Arango J, Mathieu H, Barbezier N, Anton PM. Intestinal epithelial co-culture sensitivity to pro-inflammatory stimuli and polyphenols is medium-independent. 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Bioactive Silver Nanoparticles Coated with Curcumin Inhibit Angiogenesis through MMP-9 and Cox-2 Down-Regulation 2 2 Background: Inhibition of angiogenesis is an attractive approach in cancer therapy. Both curcumin and silver nanoparticles (AgNPs) have demonstrated anti-angiogenic properties; however, the the poor water solubility of Curcumin and the side effects of AgNPs adversely affect their activity. Methods: In this study, AgNPs coated with curcumin (Cur-AgNPs), was used to improve aqueous-phase solubility of curcumin and decrease the side effects of AgNPs. After-wards, treatment with curcumin enhanced the anti-angiogenic activity of Cur-AgNPs. The nanoparticles were synthesized as both reducing and stabilizing agents. Evaluation of anti-angiogenesis was assessed in vitro using Human Umbilical Vein Endothelial Cells (HUVECs) and in vivo through the Chorioallantoic Membrane (CAM) assay. Da-ta were analyzed by one-way ANOVA with Tukey’s multiple comparison test. Results: Synthesized Cur-AgNPs have an average diameter of 39 nm, with spherical shapes and an absorbance peak at 450 nm in the UV-visible spectrum. Cur-AgNPs showed a negative zeta potential. EDAX and FTIR confirmed the conjugation of cur-cumin with AgNPs. In vitro anti-angiogenesis assays demonstrated that Cur-AgNPs re-duced the viability of HUVECs, an Inhibitory Concentration (IC50) value of 13 µg/ml. DAPI and acridine orange/propidium iodide staining revealed a significant increase in apoptotic cells following treatment with Cur-AgNPs. The expression of Matrix Metallo-proteinase 9 (MMP-9) and Cyclooxygenase-2 (COX-2) was also inhibited in treated cells. In vivo anti-angiogenesis assays using the CAM model showed significant de-crease in the number, length and hemoglobin content of CAM blood vessels. Conclusion: Curcumin conjugated with AgNPs may represent a promising strategy to enhance the therapeutic potential of both AgNPs and curcumin. However, further in-vestigations, particularly regarding safety and biocompatibility of Cur-AgNPs, is needed in this field. 147 156 Tayebe Ramezani Farzin Mohammad Nabiuni Department of Biology, Kharazmi University Department of Biology, Kharazmi University Iran AngiogenesisCurcuminNanoparticlesSilver 70651.pdf Kubota Y. Tumor angiogenesis and anti-angiogenic therapy. Keio J Med 2012;61(2):47-56. ##Ghasemali S, Farajnia S, Barzegar A, Rahmati-Yamchi M, Baghban R, Rahbarnia L, et al. New Developments in Anti-Angiogenic Therapy of Cancer, Review and Update. Anticancer Agents Med Chem 2021;21(1):3-19.##Carmeliet P. Angiogenesis in life, disease and medicine. Nature 2005 Dec 15;438(7070):932-6.##Su J, Zhu HL, Yao Y, Duan Y. Antiangiogenic therapy: challenges and future directions. Ctmc 2021 Jan 1;21:87-9.##Bhandarkar SS, Arbiser JL. Curcumin as an inhibitor of angiogenesis. Adv Exp Med Biol 2007;595:185-95.##Bhatia M, Bhalerao M, Cruz-Martins N, Kumar D. 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Identification of a Novel Nucleic Acid Target for the Rapid and Specific Detection of Mycobacterium Simiae Using Comparative Genomic Analysis 2 2 Background: Mycobacterium simiae (M. simiae) is a non-tuberculous mycobacterium (NTM) that closely resembles Mycobacterium tuberculosis (M. tuberculosis) in clinical and biochemical characteristics, notably its niacin-positive phenotype. This similarity frequently leads to misdiagnosis and inappropriate treatment with first-line anti-tuberculosis drugs, to which M. simiae is frequently resistant. Current diagnostic methods are expensive or need complex equipment, highlighting the urgent need for a rapid, specific, and accessible molecular target to identify M. simiae accurately. Methods: In this study, a modified genome comparison method was applied to the complete reference genome of M. simiae (AP022568.1) in order to identify a putative species-specific nucleotide sequence. A conventional PCR assay was designed to amplify a 168-bp fragment within this target, designated MST601 (M. simiae Target, 601 bp). The analytical sensitivity [Limit of Detection (LOD)] was determined using serial dilutions of genomic DNA. The pilot evaluation of the assay was assessed using 10 well-characterized clinical isolates of M. simiae. Results: The MST601-PCR assay demonstrated high analytical sensitivity, with a limit of detection of ~10 fg (≈2 genome equivalents per reaction) of M. simiae genomic DNA. No cross-reactivity among the tested species was observed with any of the 10 non-target mycobacterial species tested. The assay successfully amplified the target sequence from all 10 clinical isolates. Sequencing of the amplicons revealed ≥99% identity to reference M. simiae strains in the GenBank database, validating the assay's accuracy. Conclusion: A species-specific nucleic acid target, MST601, facilitating the rapid and accurate detection of M. simiae via conventional PCR was presented. This assay pro-vides a low-cost and accessible option for diagnostic laboratories. 157 166 Reza Kamali Kakhki Mashhad Gene Azma Inc. Mashhad Gene Azma Inc. Iran Mohammad Abavisani Student Research Committee, Mashhad University of Medical Sciences Student Research Committee, Mashhad University of Medical Sciences Iran Kiarash Ghazvini Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences Iran Hosna Zare Student Research Committee, Mashhad University of Medical SciencesDepartment of Laboratory Sciences, School of Paramedical Sciences, Mashhad University of Medical Sciences Student Research Committee, Mashhad University of Medical SciencesDepartment of Laboratory Sciences, School of Paramedical Sciences, Mashhad University of Medical Sciences IranIran Noshin Hojatpanah Student Research Committee, Mashhad University of Medical SciencesDepartment of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences Student Research Committee, Mashhad University of Medical SciencesDepartment of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences IranIran Jamal Falahi Department of Laboratory Sciences, School of Paramedical Sciences, Torbat Heydariyeh University of Medical Sciences Department of Laboratory Sciences, School of Paramedical Sciences, Torbat Heydariyeh University of Medical Sciences Iran Alireza Neshani Student Research Committee, Mashhad University of Medical Sciences Student Research Committee, Mashhad University of Medical Sciences Iran Molecular diagnosticsMST601Mycobacterium simiaeNovel targetPCRSensitivitySpecificity 70652.pdf Chai J, Han X, Mei Q, Liu T, Walline JH, Xu J, et al. 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