Prediction of underweight, short stature, and microcephaly based on brain diffusion-weighted imaging sequence in neonates with stage.2 of hypoxic-ischemic encephalopathy: A follow-up study پیش‌بینی کم‌وزنی، کوتاه‌قدی و میکروسفالی بر اساس توالی تصویربرداری d‌iffusion weighted imaging مغز در نوزادان با مرحله 2 انسفالوپاتی هیپوکسیک-ایسکمیک: یک مطالعه از نوع پیگیری 2 1 مقدمه: کاهش اکسیژنرسانی و جریان خون مغز طبق تعریف انسفالوپاتی هیپوکسیک-ایسکمیک (HIE) در 1 تا 8 تولد از هر 1000 تولد کامل در کشورهای توسعهیافته و تا 26 مورد در هر 1000 در کشورهای در حال توسعه رخ میدهد. هدف: وضعیت رشد بازماندگان تولد HIE به اندازه کافی ارزیابی نشده است. در این مطالعه پارامترهای رشد (وزن، قد و دور سر) نوزادان مبتلا به سرنات مرحله 2 HIE در ماه‌های 6، 10 و 12 و ارتباط آن با یافته‌های تصویربرداری وزنی با انتشار مغز نوزادان (DWI) مورد بررسی قرار گرفت. مواد و روش ها: پارامترهای رشد 35 نوزاد با سن حاملگی بالاتر از 34 هفته که با مرحله 2 HIE در بخش مراقبت‌های ویژه نوزادان بیمارستان شهید صدوقی یزد از اسفند 1399 تا اسفند 1400 بستری شده بودند، از پرونده بیماران استخراج شد. ارتباط با یافتههای توالی DWI مغز نوزادان ارزیابی شد. نتایج: 15 دختر و 20 پسر با میانگین وزن هنگام تولد 8/221 ± 3/2880 گرم مورد بررسی قرار گرفتند. تصویربرداری رزونانس مغناطیسی معمولی (MRI) و DWI در 6 نوزاد (1/%17) و 18 نوزاد (4/%51) غیرطبیعی بود. غیرطبیعیترین یافته DWI سیگنال بالا در گانگلیون قاعده/تالاموس در 9 نوزاد (7/%25) بود. DWI غیرطبیعی در نوزادان مبتلا به تشنج و در نوزادان کموزن بیشتر است. روزهای بستری در نوزادان با DWI غیرطبیعی طولانیتر بود. میکروسفالی در 12 ماهگی در کودکان مبتلا به DWI غیرطبیعی بیشتر بود. نتیجه گیری: در بازماندگان HIE نوزادان متوسط، توالی غیرطبیعی DWI مغز ممکن است رشد نامناسب سر را پیش‌بینی کند و برای بهبود رشد نیاز به مداخلات پزشکی و تغذیه‌ای دقیق دارند. 2 Background: Hypoxic-ischemic encephalopathy (HIE), caused due to reduced oxygenation and brain blood flow, occurs in 1-8 per 1000 live full-term births in developed countries and up to 26 per 1000 live in the developing world. The growth status of survivors of birth HIE has not been evaluated sufficiently. Objective: This study evaluated, the growth parameters (weight, height, and head circumference) of neonates with Sarnat stage.2 of HIE at 6, 10, and 12 months and its relationship with findings of neonatal brain diffusion-weighted imaging (DWI) sequence. Materials and Methods: Medical records and growth parameters of 35 neonates with gestational age > 34 wk who were admitted with stage.2 of HIE in Neonatal Intensive Care Unit of Shahid Sadoughi hospital, Yazd, Iran from March 2021-March 2022, and its relationship with neonatal brain DWI sequence finding was evaluated. Results: 15 girls and 20 boys with a mean birth weight of 2880.3 ± 221.8 gr were evaluated. Conventional magnetic resonance imaging and DWI were found to be abnormal in 6 (17.1%) and 18 neonates (51.4%). The most abnormal finding of DWI was high signal in basal ganglia/thalamus in 9 neonates (25.7%). Abnormal DWI is more frequent in neonates with seizures and low birth weight. Hospital stay days were more prolonged in neonates with abnormal DWI. Microcephaly at 12 months was more frequent in children with abnormal DWI. Conclusion: In survivors of moderate neonatal HIE, abnormal brain DWI sequence might predict inappropriate head growth, and need close medical and nutritional interventions for growth improvement. 1 8 2023/06/26 1402/4/5 2023/12/25 1402/10/4 Mohammad Golshan Tafti Mohammad Golshan Tafti Department of Pediatrics, Ali-ebn-Abitaleb School of Medicine, Islamic Azad University, Yazd Branch, Yazd, Iran. ایران mgolshan035@gmail.com Marjan Jafari Marjan Jafari Department of Neonatology, Shahid Beheshti Hospital, Isfahan University of Medical Sciences, Isfahan, Iran. ایران dr.marjanjafari@gmail.com Seyed Reza Mirjalili Seyed Reza Mirjalili Department of Neonatology, Mother and Newborn Health Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران dr_sr_mirjalili@yahoo.com Razieh Fallah Razieh Fallah Department of Pediatrics, Children Growth Disorder Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران dr.raziehfallah@yahoo.com Farimah Shamsi Farimah Shamsi Center for Healthcare Data Modeling, Department of Biostatistics and Epidemiology, School of Public Health, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران farimahshamssi@gmail.com Hypoxic ischemia encephalopathy Magnetic resonance imaging Diffusion weighted imaging Microcephaly Underweight. هایپوکسیک ایسکمیک انسفالوپاتی تصویربرداری رزونانس مغناطیسی تصویربرداری با وزن انتشار میکروسفالی کم وزنی. 1. El-Mazary AAM, Nasif KhA, Abdel-Hakeem GL, Sherif T, Farouk E, El-Gezawy EM. Adiponectin, leptin and insulin levels at birth and in early postnatal life in neonates with hypoxic ischemic encephalopathy. J Diabetes Metab Disord 2015; 14: 87.##2. Groenendaal F, de Vries LS. Fifty years of brain imaging in neonatal encephalopathy following perinatal asphyxia. Pediatr Res 2017; 81: 150-155.##3. Bano Sh, Chaudhary V, Garga UCh. Neonatal hypoxic-ischemic encephalopathy: A radiological review. J Pediatr Neurosci 2017; 12: 1-6.##4. Rana L, Sood D, Chauhan R, Shukla R, Gurnal P, Nautiyal H, et al. MR imaging of hypoxic ischemic encephalopathy - distribution patterns and ADC value correlations. Eur J Radiol Open 2018; 5: 215-220.##5. Gunn AJ, Thoresen M. Neonatal encephalopathy and hypoxic-ischemic encephalopathy. Handb Clin Neurol 2019; 162: 217-237.##6. Walas W, Wilińska M, Bekiesińska-Figatowska M, Halaba Z, Śmigiel R. Methods for assessing the severity of perinatal asphyxia and early prognostic tools in neonates with hypoxic-ischemic encephalopathy treated with therapeutic hypothermia. Adv Clin Exp Med 2020; 29: 1011-1016.##7. Cascio A, Ferrand A, Racine E, St-Hilaire M, Sanon PN, Gorgos A, et al. Discussing brain magnetic resonance imaging results for neonates with hypoxic-ischemic encephalopathy treated with hypothermia: A challenge for clinicians and parents. eNeurologicalSci 2022; 29: 100424.##8. Fallah R, Akhavan Karbasi S, Galalian MT, Dehghani-Firouzabadi R. Comparison of developmental status of 5-year-old singleton children born through assisted and natural conceptions. Iran J Reprod Med 2013; 11: 365-370.##9. Karimi M, Fallah R, Dehghanpoor A, Mirzaei M. Developmental status of 5-year-old moderate low birth weight children. Brain Dev 2011; 33: 651-655.##10. Islami Z, Fallah R, Mosavian T, Pahlavanzadeh MR. Growth parameters of NICU admitted low birth weight preterm neonates at corrected ages of 6 and 12 month. Iran J Reprod Med 2012; 10: 459-464.##11. Ashwal S, Michelson D, Plawner L, Dobyns WB, Quality Standards Subcommittee of the American Academy of Neurology and the Practice Committee of the Child Neurology Society. Practice parameter: Evaluation of the child with microcephaly (an evidence-based review): Report of the Quality Standards Subcommittee of the American Academy of Neurology and the Practice Committee of the Child Neurology Society. Neurology 2009; 73: 887-897.##12. Shankaran S, Pappas A, McDonald SA, Vohr BR, Hintz SR, Yolton K, et al. Childhood outcomes after hypothermia for neonatal encephalopathy. N Engl J Med 2012; 366: 2085-2092.##13. Ristovska S, Stomnaroska O, Danilovski D. Hypoxic ischemic encephalopathy (HIE) in term and preterm infants. Pril 2022; 43: 77-84.##14. Guillet R, Edwards AD, Thoresen M, Ferriero DM, Gluckman PD, Whitelaw A, et al. Seven- to eight-year follow-up of the CoolCap trial of head cooling for neonatal encephalopathy. Pediatr Res 2012; 71: 205-209.##15. Preeti S, Kadam A, Kadam S, Vaidya U, Kumar P, Bhagat I, et al. Anthropometric measures as biomarkers of neurodevelopmental outcomes of newborns with moderate to severe hypoxic ischemic encephalopathy. J Neonatal Perinatal Med 2019; 12: 127-134.##16. Murden S, Borbélyová V, Laštůvka Z, Mysliveček J, Otáhal J, Riljak V. Gender differences involved in the pathophysiology of the perinatal hypoxic-ischemic damage. Physiol Res 2019; 68 (Suppl.): S207-S217.##17. Russ JB, Simmons R, Glass HC. Neonatal encephalopathy: Beyond hypoxic-ischemic encephalopathy. Neoreviews 2021; 22: e148-e162.##18. Kwon JM, Guillet R, Shankaran S, Laptook AR, McDonald SA, Ehrenkranz RA, et al. Clinical seizures in neonatal hypoxic-ischemic encephalopathy have no independent impact on neurodevelopmental outcome: Secondary analyses of data from the neonatal research network hypothermia trial. J Child Neurol 2011; 26: 322-328.## ##

Intrauterine autologous platelet-rich plasma treatment in women with at least two implantation failures: A retrospective cohort study درمان پلاسمای غنی از پلاکت اتولوگ داخل رحمی در زنان با حداقل دو شکست لانه-گزینی: یک مطالعه کوهورت گذشته نگر 2 1 مقدمه: یافتن مؤثرترین راه برای بهبود میزان لانهگزینی در زنانی که تحت درمان با تکنولوژی کمک باروری هستند هنوز یک چالش است. هدف: این مطالعه با هدف ارزیابی پیامدهای حاملگی درمان با پلاسمای غنی از پلاکت داخل رحمی (PRP) در زنان با سابقه حداقل دو شکست لانهگزینی انجام شد. مواد و روش ها: در این مطالعه کوهورت گذشته‌نگر، داده‌های 852 زن کاندید انتقال جنین منجمد-ذوب شده از فروردین 1396 تا شهریور 1400 در پژوهشکده علوم تولیدمثل یزد ایران استخراج شد. از این تعداد، 432 مورد درمان PRP داخل رحمی 48 ساعت قبل از انتقال دریافت کردند (گروه PRP) و نتایج بارداری با 420 نفر (گروه کنترل) که درمان را قبل از انتقال دریافت نکردند مقایسه شد. نتایج: پیامدهای حاملگی شامل میزان حاملگی شیمیایی،کلینیکی، بارداری در حال پیشرفت و تولد زنده در گروه PRP از نظر آماری تفاوت معنی‌دار داشت (001/0 > p). اگرچه هنگامی که بر اساس تاریخچه تعداد موارد شکست لانهگزینی طبقهبندی انجام شد، این بهبود قابل توجه در هر چهار مورد تنها در زنان با سابقهی حداقل دو شکست لانهگزینی دیده شد. در زنان با سابقه تنها یک شکست لانهگزینی، درمان با پیآرپی به طور قابل توجهی باعث بهبود حاملگی در حال پیشرفت و میزان تولد زنده شد (5/19%، 04/0 = p). همچنین، در زنانی که تخمک اهدایی دریافت کردند و شکست لانهگزینی مکرر داشتند، PRP باعث بهبود بارداری شد. اما از نظر آماری معنیدار نبود (15/0 = p). نتیجه گیری: به نظر می‌رسد PRP در بهبود میزان بارداری در زنان با سابقه 2 یا بیشتر شکست لانه‌گزینی مؤثر بوده و همچنین افزایش نرخ تولد زنده را در بیمارانی که تنها یک بار لانه‌گزینی ناموفق داشتند نشان می‌دهد. 2 Background: Finding the most effective way to improve implantation rate in women who are receiving assisted reproductive technology treatment is still a challenge. Objective: This study aimed to assess the pregnancy outcomes of intrauterine platelet-rich plasma (PRP) therapy in women with a history of at least 2 implantation failures. Materials and Methods: In this retrospective cohort study, data of 852 women who were candidates for frozen-thawed embryo transfer was extracted from their medical records from April 2017 to September 2021 at Yazd Reproductive Sciences Institute, Yazd, Iran. Of these, 432 received intrauterine PRP treatment 48 hr before transfer (PRP group), and the results of the pregnancy outcomes compared with 420 of the control group who did not receive the treatment before transfer. Results: Pregnancy outcomes, including chemical, clinical, ongoing pregnancy, and live birth rate were statistically significant in the PRP group (p < 0.001). However, when categorized according to the implantation history, this significant improvement in all 4 was only seen in women with at least 2 prior implantation failures. In women with a history of only one implantation failure, PRP therapy significantly improved the ongoing pregnancy and live birth rate (19.5%, p = 0.04). Also, in women who received donor eggs and had repeated implantation failure, PRP improved pregnancy outcomes clinically but not statistically (p = 0.15). Conclusion: PRP seems to be effective in improving the pregnancy rate in women with a history of 2 or more implantation failures and also shows an increase in the live birth rate in women with only one implantation failure. 9 16 2023/06/262023/10/24 1402/8/2 2023/12/252023/12/19 1402/9/28 Neda Fattahi Meybodi Neda Fattahi Meybodi Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران nedafatahi@gmail.com Maryam Eftekhar Maryam Eftekhar Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران eftekharmaryam@yahoo.com Behnaz Gandom Behnaz Gandom Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران behnaz.gandom@yahoo.com Platelet-rich plasma Embryo implantation Assisted reproductive technology Ovum donor Live birth rate. پلاسمای غنی از پلاکت شکست لانه‌گزینی جنین فناوری کمک باروری اهداکننده تخمک میزان تولد زنده. 1. Nazari L, Salehpour S, Hoseini S, Zadehmodarres Sh, Ajori L. Effects of autologous platelet-rich plasma on implantation and pregnancy in repeated implantation failure: A pilot study. Int J Reprod BioMed 2016; 14: 625-628.##2. Kasius A, Smit JG, Torrance HL, Eijkemans MJ, Mol BW, Opmeer BC, et al. Endometrial thickness and pregnancy rates after IVF: A systematic review and meta-analysis. Hum Reprod Update 2014; 20: 530-541.##3. Aghajanova L, Houshdaran S, Balayan S, Irwin J, Huddleston H, Giudice L. Platelets for endometrial regeneration: A novel approach. Fertil Steril 2016; 106: e82.##4. Arefi S, Fazeli E, Esfahani M, Borhani N, Yamini N, Hosseini A, et al. Granulocyte-colony stimulating factor may improve pregnancy outcome in patients with history of unexplained recurrent implantation failure: An RCT. Int J Reprod BioMed 2018; 16: 299-304.##5. Barad DH, Yu Y, Kushnir VA, Shohat-Tal A, Lazzaroni E, Lee H-J, et al. A randomized clinical trial of endometrial perfusion with granulocyte colony-stimulating factor in in vitro fertilization cycles: Impact on endometrial thickness and clinical pregnancy rates. Fertil Steril 2014; 101: 710-715.##6. Bos-Mikich A, Ferreira MO, de Oliveira R, Frantz N. Platelet-rich plasma or blood-derived products to improve endometrial receptivity? J Assist Reprod Genet 2019; 36: 613-620.##7. Hviid MM, Macklon N. Immune modulation treatments-where is the evidence? Fertil Steril 2017; 107: 1284-1293.##8. Jain S, Mahey R, Malhotra N, Kalaivani M, Sangeeta P, Bhatt A, et al. Effect of intrauterine perfusion of granulocyte colony-stimulating factor on endometrial parameters and in vitro fertilization outcome in women undergoing in vitro fertilization/intracytoplasmic sperm injection cycles: A randomized controlled trial. J Hum Reprod Sci 2018; 11: 254-260.##9. Tandulwadkar SR, Naralkar MV, Surana AD, Selvakarthick M, Kharat AH. Autologous intrauterine platelet-rich plasma instillation for suboptimal endometrium in frozen embryo transfer cycles: A pilot study. J Hum Reprod Sci 2017; 10: 208-212.##10. Davari-Tanha F, Tehraninejad ES, Ghazi M, Shahraki Z. The role of G-CSF in recurrent implantation failure: A randomized double blind placebo control trial. Int J Reprod BioMed 2016; 14: 737-742.##11. Huang C, Ye X, Ye L, Lu L, Liu F. Platelet-rich plasma intrauterine infusion as assisted reproduction technology (ART) to combat repeated implantation failure (RIF): A systematic review and meta-analysis. Iran J Public Health 2023; 52: 1542-1554.##12. Welte K. G-CSF: Filgrastim, lenograstim and biosimilars. Expert Opin Biol Ther 2014; 14: 983-993.##13. Zadehmodarres Sh, Salehpour S, Saharkhiz N, Nazari L. Treatment of thin endometrium with autologous platelet-rich plasma: A pilot study. JBRA Assist Reprod 2017; 21: 54-56.##14. Eftekhar M, Neghab N, Khani P. Effectiveness of autologous platelet-rich plasma therapy in women with repeated implantation failure: An RCT. Int J Fertil Steril 2023; in Press.##15. Eftekhar M, Neghab N, Naghshineh E, Khani P. Can autologous platelet rich plasma expand endometrial thickness and improve pregnancy rate during frozen-thawed embryo transfer cycle? A randomized clinical trial. Taiwan J Obstet Gynecol 2018; 57: 810-813.##16. Tehraninejad ES, Kashani NG, Hosseini A, Tarafdari A. Autologous platelet‐rich plasma infusion does not improve pregnancy outcomes in frozen embryo transfer cycles in women with history of repeated implantation failure without thin endometrium. J Obstet Gynaecol Res 2021; 47: 147-151.##17. Ban Y, Yang X, Xing Y, Que W, Yu Z, Gui W, et al. Intrauterine infusion of leukocyte-poor platelet-rich plasma is an effective therapeutic protocol for patients with recurrent implantation failure: A retrospective cohort study. J Clin Med 2023; 12: 2823.##18. Xu Y, Hao C, Fang J, Liu X, Xve P, Miao R. Intrauterine perfusion of autologous platelet-rich plasma before frozen-thawed embryo transfer improves the clinical pregnancy rate of women with recurrent implantation failure. Front Med 2022; 9: 64-67.##19. Kong X, Tang G, Liu Y, Zheng Z, Li Y, Yan F. Efficacy of intrauterine infusion therapy before embryo transfer in recurrent implantation failure: A systematic review and network meta-analysis. J Reprod Immunol 2023; 156: 103819.##20. Deng H, Wang S, Li Z, Xiao L, Ma L. Effect of intrauterine infusion of platelet-rich plasma for women with recurrent implantation failure: A systematic review and meta-analysis. J Obstet Gynaecol 2023; 43: 2144177.##21. Kaur H, Meenu M, Pandey S, Chauhan A, Mangla M. Efficacy and safety of PRP in patients with recurrent implantation failure undergoing ART-A systematic review & meta-analysis. Authorea Preprints; 2023.##22. Abd Elsalam Elgendy HE, Mohamed SA. Efficacy of intra-uterine infusion of PRP for pregnancy related outcomes in women with recurrent implantation failure; systematic review and meta-analysis of published trials. Egypt J Fertil Steril 2023; 27: 18-26.## ##

Sertoli cell-conditioned medium can improve blood-testis-barrier function and spermatogenesis in azoospermia mice induced by scrotal hyperthermia: An experimental study محیط کشت بهینه مشتق از سلول های سرتولی می تواند سبب بهبود عملکرد سد خونی-بیضه ای و اسپرماتوژنز در موش های مدل آزواسپرمی القا شده با هیپرترمی اسکروتال شود: یک مطالعه تجربی 2 1 مقدمه: افزایش دمای بیضه با آسیب به اپیتلیوم لولههای اسپرمساز و اختلال در تولید اسپرم همراه است. هدف: هدف از مطالعه کنونی بررسی اثرات محیط کشت بهینه مشتق از سلولهای سرتولی (SCCM) بر ژنهای مرتبط با سد خونی-بیضهای و فرآیند اسپرماتوژنز به دنبال القای هایپرترمی اسکروتال بود. مواد و روش ها: در این مطالعه 40 سر موش سوری نژاد NMRI بالغ 8 هفته با وزن 30-25 گرم بهصورت تصادفی به 4 گروه تقسیم شدند: (I کنترل، DMEM (II، (III هایپرترمی اسکروتال، و IV) هایپرترمی اسکروتال+SCCM. هایپرترمی با قرار دادن ناحیه اسکروتال موشها در آب 43 درجه سانتیگراد به مدت 20 دقیقه به صورت یک روز در میان و به مدت 10 روز القا گردید. موشها در گروه II و IV به مدت 5 هفته و به صورت یک روز درمیان به ترتیب (μl 10) DMEM F/12 و (μl 10) SCCM دریافت کردند. سپس موشها قربانی شده و دم اپی-دیدیم آنها جهت بررسی پارامترهای اسپرم، بیضهها جهت مطالعات استریولوژیکال، سطح ROS و GSH، و بیان ژنهای Ocln, Gja1, Cdh2 و Itgb1 جدا شدند. نتایج: نتایج آنالیز اسپرم نشان داد که در موشهای تحت درمان با SCCM بهصورت معناداری تعداد و تحرک اسپرمها افزایش و از همگسیختگی DNA آنها کاهش یافته بود. همچنین، یافتههای بافتی و مولکولی نشاندهنده افزایش حجم بافت بیضه، شمار سلولهای زایا، سطح GSH و بیان ژنهای Ocln, Gja1, Cdh2 و Itgb1 در گروه تحت درمان با SCCM در مقایسه با گروه هایپرترمی اسکروتال بود. نتیجه گیری: یافته‌ها نشان می‌دهد که فاکتورهای رشد SCCM تکثیر و تمایز سلول‌های زایا را از طریق اثرات پاراکرین تحریک می‌کنند و بیان ژنهای مرتبط با سد خونی-بیضهای را در موشهای تحت القای هایپرترمی اسکروتال تنظیم میکنند. 2 Background: An increase in the temperature of the testis is associated with damage to the epithelium of seminiferous tubules and disruption of sperm production. Objective: The current study aimed to investigate the effect of the Sertoli cell-conditioned medium (SCCM) on the blood-testis-barrier associated genes and spermatogenesis process following scrotal hyperthermia. Materials and Methods: In this experimental study, 40 adult NMRI mice (8 wk, 25-30 gr) were allocated into 4 groups: I) control, II) DMEM (10 μl Dulbecco's Modified Eagle Medium), III) scrotal hyperthermia, and IV) scrotal hyperthermia+SCCM (10 μl SCCM). Hyperthermia was induced by placing the mice scrotum in water at 43°C for 20 min every other day for 10 days. Mice were treated every other day for 5 wk. Then the animals were euthanized, and the tails of epididymis were removed to analyze sperm parameters, testis were taken for stereological assessment, reactive oxygen spices and glutathione levels, and the expression of Ocln, Gja1, Cdh2, and Itgb1. Results: The results of sperm analysis indicated that SCCM-treated mice significantly increased sperm count and motility and reduced DNA fragmentation. In addition, histological and molecular findings showed that the volume of testicular tissue, the number of germ cells, the glutathione level, and the expression of Ocln, Gja1, Cdh2, and Itgb1 genes were significantly increased in the SCCM-treated mice. Conclusion: Findings suggest that growth factors of SCCM stimulate the proliferation and differentiation of germ cells through paracrine effects and upregulate the blood-testis-barrier-associated genes in mice subjected to scrotal hyperthermia. 17 30 2023/06/262023/10/242023/10/26 1402/8/4 2023/12/252023/12/192024/01/8 1402/10/18 Fakhroddin Aghajanpour Fakhroddin Aghajanpour Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران fakhroddin72@gmail.com Reza Soltani Reza Soltani Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران rezasoltanikazemii@gmail.com Azar Afshar Azar Afshar Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Student Research Committee, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران azar.afshar8894@gmail.com Hojjat-Allah Abbaszadeh Hojjat-Allah Abbaszadeh Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران hoomanabs@gmail.com Fatemeh Fadaei Fathabadi Fatemeh Fadaei Fathabadi Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران fatemeh.fadaeifathabadi@gmail.com Nafiseh Moeinian Nafiseh Moeinian Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران moeinian.ptu@gmail.com Abbas Aliaghaei Abbas Aliaghaei Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران aghaei60@gmail.com Ali Dehghani Nejad Ali Dehghani Nejad Department of Biology and Anatomical Sciences, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran. ایران amirali3532@gmail.com Reza Mastery Farahani Reza Mastery Farahani Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران realmastery@hotmail.com Mohsen Norouzian Mohsen Norouzian Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران norozian93@gmail.com Mohammad-Amin Abdollahifar Mohammad-Amin Abdollahifar Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ایران m_amin58@yahoo.com Testis Sertoli cells Culture media Hyperthermia Spermatogenesis. بیضه سلول سرتولی محیط کشت هیپرترمی اسپرماتوژنز. 1. McGregor GR, Vanos JK. Heat: A primer for public health researchers. Public Health 2018; 161: 138-146.##2. Kastelic JP, Rizzoto GJBr. Thermoregulation of the testes. In: Hopper RM. Bovine reproduction. 2th Ed. US: John Wiley & Sons; 2021. ##3. Abd El-Emam MM, Ray MN, Ozono M, Kogure K. Heat stress disrupts spermatogenesis via modulation of sperm-specific calcium channels in rats. J Therm Biol 2023; 112: 103465.##4. Durairajanayagam D, Agarwal A, Ong C. Causes, effects and molecular mechanisms of testicular heat stress. Reprod Biomed Online 2015; 30: 14-27.##5. Mäkelä J-A, Koskenniemi JJ, Virtanen HE, Toppari J. Testis development. Endocr Rev 2019; 40: 857-905.##6. O’Donnell L, Smith LB, Rebourcet D. Sertoli cells as key drivers of testis function. Semin Cell Dev Biol 2022; 121: 2-9.##7. Arisha AH, Ahmed MM, Kamel MA, Attia YA, Hussein MMA. Morin ameliorates the testicular apoptosis, oxidative stress, and impact on blood–testis barrier induced by photo-extracellularly synthesized silver nanoparticles. Environ Sci Pollut Res Int 2019; 26: 28749-28762.##8. Hu Y, Hu H, Yin L, Wang L, Luo K, Luo N, et al. Arachidonic acid impairs the function of the blood-testis barrier via triggering mitochondrial complex-ROS-P38 MAPK axis in hyperthermal Sertoli cells. Ecotoxicol Environ Saf 2023; 252: 114598.##9. Mruk DD, Cheng CY. The mammalian blood-testis barrier: Its biology and regulation. Endocr Rev 2015; 36: 564-591.##10. Chen S-R, Liu Y-X. Regulation of spermatogonial stem cell self-renewal and spermatocyte meiosis by Sertoli cell signaling. Reproduction 2015; 149: R159-R167.##11. Hai Y, Hou J, Liu Y, Liu Y, Yang H, Li Z, et al. The roles and regulation of Sertoli cells in fate determinations of spermatogonial stem cells and spermatogenesis. Semin Cell Dev Biol 2014; 29: 66-75.##12. Hajian Monfared M, Minaee B, Rastegar T, Khrazinejad E, Barbarestani M. Sertoli cell condition medium can induce germ like cells from bone marrow derived mesenchymal stem cells. Iran J Basic Med Sci 2016; 19: 1186-1192.##13. Panahi S, Karamian A, Sajadi E, Aliaghaei A, Nazarian H, Abdi Sh, et al. Sertoli cell-conditioned medium restores spermatogenesis in azoospermic mouse testis. Cell Tissue Res 2020; 379: 577-587.##14. Afshar A, Aliaghaei A, Nazarian H, Abbaszadeh H-A, Naserzadeh P, Fathabadi FF, et al. Curcumin-loaded iron particle improvement of spermatogenesis in azoospermic mouse induced by long-term scrotal hyperthermia. Reprod Sci 2021; 28: 371-380.##15. Ziaeipour S, Piryaei A, Aliaghaei A, Nazarian H, Naserzadeh P, Ebrahimi V, et al. Chronic scrotal hyperthermia induces azoospermia and severe damage to testicular tissue in mice. Acta Histochem 2021; 123: 151712.##16. Ayoubi M, Naserzadeh P, Hashemi MT, Rostami MR, Tamjid E, Tavakoli MM, et al. Biochemical mechanisms of dose-dependent cytotoxicity and ROS-mediated apoptosis induced by lead sulfide/graphene oxide quantum dots for potential bioimaging applications. Sci Rep 2017; 7: 12896.##17. Oliveira PF, Alves MG. Sertoli cell metabolism and spermatogenesis. Cham: Springer; 2015: 25-39. ##18. Parekh PA, Garcia TX, Hofmann M-C. Regulation of GDNF expression in Sertoli cells. Reproduction 2019; 157: R95-R107.##19. Peng YJ, Tang XT, Shu HS, Dong W, Shao H, Zhou BO. Sertoli cells are the source of stem cell factor for spermatogenesis. Development 2023; 150: dev200706.##20. Meroni SB, Galardo MN, Rindone G, Gorga A, Riera MF, Cigorraga SB. Molecular mechanisms and signaling pathways involved in sertoli cell proliferation. Front Endocrinol 2019; 10: 224.##21. Muratori M, Tamburrino L, Marchiani S, Cambi M, Olivito B, Azzari C, et al. Investigation on the origin of sperm DNA fragmentation: Role of apoptosis, immaturity and oxidative stress. Mol Med 2015; 21: 109-122.##22. Fu L, Yuen KCJ, Tint AN, Hoffman AR, Bongso AT, Lee KO. Association of decreased sperm motility and increased seminal plasma IGF-I, IGF-II, IGFBP-2, and PSA levels in infertile men. Endocrine 2021; 74: 698-706.##23. Ghafouri-Fard S, Shoorei H, Mohaqiq M, Raza SHA, Taheri M. The role of different compounds on the integrity of blood-testis barrier: A concise review based on in vitro and in vivo studies. Gene 2021; 780: 145531.##24. Su L, Wang Z, Xie S, Hu D, Cheng YC, Mruk DD, et al. Testin regulates the blood‐testis barrier via disturbing occludin/ZO‐1 association and actin organization. J Cell Physiol 2020; 235: 6127-6138.##25. Piprek RP, Kloc M, Mizia P, Kubiak JZ. The central role of cadherins in gonad development, reproduction, and fertility. Int J Mol Sci 2020; 21: 8264.##26. Zhang DC, Chen R, Cai YH, Wang JJ, Yin C, Zou K. Hyperactive reactive oxygen species impair function of porcine Sertoli cells via suppression of surface protein ITGB1 and connexin-43. 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Modulating the RFamide-related peptide-3/G protein-coupled receptor 147 signaling pathway with nourishing Yin-removing fire herbal mixture to alleviate precocious puberty in female rats: An experimental study تعدیل مسیر سیگنال دهی گیرنده 147 متصل به پروتئین پپتید-3/G مرتبط با RFamide با تجویز مخلوط گیاهی Yin-removing fire برای کاهش بلوغ زودرس موش های صحرایی ماده: یک مطالعه تجربی 2 1 مقدمه: بلوغ زودرس (PP) شامل فعال شدن زودهنگام مولد هورمون آزادکننده گنادوتروپین هیپوتالاموس (GnRH) است. مسیر سیگنالدهی گیرنده 147 متصل به پروتئین پپتید-3/G مرتبط با RFamide (RFRP3/GPR147) در مهار GnRH و به تأخیر انداختن شروع بلوغ حیاتی است. مخلوط گیاهی Yin-removing fire (NYRF) نتایج امیدوارکنندهای را در درمان PP نشان داده است. هدف: این مطالعه با هدف ارزیابی تأثیر مخلوط گیاهی NYRF بر مسیر سیگنالینگ RFRP3/GPR147 در هیپوتالاموس و توانایی آن در کاهش PP در موش‌های ماده انجام شد. مواد و روش ها: در یک آزمایش کنترل ‌شده، 24 موش صحرایی ماده نژاد Sprague-Dawley (69/0 ± 20/11 گرم، 5 روز پس از زایمان [PD5]) به گروه‌های نرمال، مدل، نرمال سالین و NYRF (هر کدام 6 رأس) تقسیم شدند. PP در گروههای مدل، نرمال سالین و NYRF با تزریق زیر جلدی دانازول در PD5 القا شد. مخلوط گیاهی NYRF یا نرمال سالین از PD15 تجویز شد. سطح سرمی هورمونهای جنسی و نمونههای هیپوتالاموس برای بررسی بیان mRNA و پروتئین در PD30 جمعآوری شد. نتایج: در گروه مدل، سطوح GnRH هیپوتالاموس و کیسپپتین افزایش یافت، در حالی که سطوح RFRP3 و GPR147 کاهش یافت، سطح هورمون لوتئینهکننده افزایش یافت، ضرایب اندام تناسلی افزایش یافت و واژن نسبت به گروه عادی زودتر باز شد. در حالیکه برعکس، گروه NYRF سطوح GnRH و کیسپپتین کمتر اما سطوح RFRP3 بالاتری را در هیپوتالاموس نشان دادند. همچنین در این گروه سطح هورمون لوتئینیزهکننده سرم کاهش یافت، ضرایب اندام تناسلی کاهش یافت و باز شدن واژن در مقایسه با مدل و گروه نرمال سالین به تعویق افتاد. نتیجه گیری: مخلوط گیاهی NYRF رشد جنسی در موش‌های مبتلا به PP را با افزایش RFRP3 هیپوتالاموس و کاهش GnRH و کیسپپتین به تأخیر انداخت. 2 Background: Precocious puberty (PP) involves early activation of the hypothalamic gonadotropin-releasing hormone (GnRH) generator. The RFamide-related peptide/G protein-coupled receptor 147 (RFRP3/GPR147) signaling pathway is vital in inhibiting GnRH and delaying puberty onset. The nourishing Yin-removing fire (NYRF) herbal mixture has shown promising results in treating PP. Objective: This study aimed to assess the impact of the NYRF herbal mixture on the RFRP3/GPR147 signaling pathway in the hypothalamus and its potential in alleviating PP in female rats. Materials and Methods: In a controlled experiment, 24 female Sprague-Dawley rats (11.20 ± 0.69 gr, postnatal day [PD5]) were divided into normal, model, normal saline, and NYRF groups (n = 6/each). PP was induced in the model, normal saline, and NYRF groups by subcutaneous injection of danazol at PD5. The NYRF herbal mixture or normal saline was administered from PD15. Serum sex hormone levels and hypothalamic samples were collected for mRNA and protein expression at PD30. Results: In the model group, hypothalamic GnRH and kisspeptin levels increased, while RFRP3 and GPR147 levels decreased, luteinizing hormone levels elevated, reproductive organ coefficients increased, and the vagina opened earlier compared to the normal group. Conversely, the NYRF group exhibited lower GnRH and kisspeptin levels but higher RFRP3 levels in the hypothalamus. Serum luteinizing hormone levels were reduced, reproductive organ coefficients were reduced, and the vaginal opening was delayed compared to the model and normal saline groups. Conclusion: The NYRF herbal mixture delayed sexual development in rats with PP by hypothalamic upregulating RFRP3 and downregulating GnRH and kisspeptin. 31 42 2023/06/262023/10/242023/10/262023/07/2 1402/4/11 2023/12/252023/12/192024/01/82024/01/13 1402/10/23 Xiaoli Dai Xiaoli Dai Traditional Chinese Medicine Department, Children's Hospital of Fudan University, Shanghai, China. چین yezi_i@163.com Yuanyuan He Yuanyuan He Traditional Chinese Medicine Department, Children's Hospital of Fudan University, Shanghai, China. چین yuanyuan_he@qq.com Suhuan Li Suhuan Li Yantaishan Hospital, Shangdong, China. چین lisuhuan1987@qq.com Yanyan Sun Yanyan Sun Traditional Chinese Medicine Department, Children's Hospital of Fudan University, Shanghai, China. چین sunyanyan09@fudan.edu.cn Wen Sun Wen Sun Traditional Chinese Medicine Department, Children's Hospital of Fudan University, Shanghai, China. چین sunwenhere@163.com Zhanzhuang Tian Zhanzhuang Tian Department of Integrative Medicine and Neurobiology, Fudan University, Shanghai, China. چین tianvv@shum.edu.cn Jian Yu Jian Yu Traditional Chinese Medicine Department, Children's Hospital of Fudan University, Shanghai, China. چین yuj@shmu.edu.cn Nurgul Yerkinkyzy Ablakimova Nurgul Yerkinkyzy Ablakimova Department of Pharmacology, West Kazakhstan Marat Ospanov Medical University, Aktobe, Kazakhstan. چین n.ablakimova@zkmu.kz Yonghong Wang Yonghong Wang Traditional Chinese Medicine Department, Children's Hospital of Fudan University, Shanghai, China. چین wyhekyy@126.com Nourishing Yin-removing fire RFamide-related peptide-3 G protein-coupled receptor 147 Hypo-thalamus Puberty Precocious. مخلوط گیاهی Yin-removing fire پپتید-3 مرتبط با RFamide گیرنده 147 متصل به پروتئین G هیپوتالاموس بلوغ زودرس. 1. Bradley SH, Lawrence N, Steele C, Mohamed Z. Precocious puberty. BMJ 2020; 368: l6597. ##2. Liu Y, Yu T, Li X, Pan D, Lai X, Chen Y, et al. Prevalence of precocious puberty among Chi-nese children: A school population-based study. Endocrine 2021; 72: 573-581. ##3. Naulé L, Maione L, Kaiser UB. Puberty, a sensitive window of hypothalamic development and plasticity. Endocrinology 2021; 162: bqaa209. ##4. Plant TM, Steiner RA. The fifty years following the discovery of gonadotropin‐releasing hor-mone. J Neuroendocrinol 2022; 34: e13141. ##5. Xie Q, Kang Y, Zhang C, Xie Y, Wang C, Liu J, et al. The role of kisspeptin in the control of the hypothalamic-pituitary-gonadal axis and reproduction. Front Endocrinol 2022; 13: 925206. ##6. Singh P, Anjum S, Srivastava RK, Tsutsui K, Krishna A. Central and peripheral neuropeptide RFRP-3: A bridge linking reproduction, nutrition, and stress response. Front Neuroendocrinol 2022; 65: 100979. ##7. Tsutsui K, Ubuka T. Discovery of gonadotropin-inhibitory hormone (GnIH), progress in GnIH research on reproductive physiology and behavior and perspective of GnIH research on neuro-endocrine regulation of reproduction. Mol Cell Endocrinol 2020; 514: 110914. ##8. Tsutsui K, Ubuka T. Gonadotropin-inhibitory hormone (GnIH): A new key neurohormone con-trolling reproductive physiology and behavior. Front Neuroendocrinol 2021; 61: 100900. ##9. Wang H, Khoradmehr A, Jalali M, Salehi MS, Tsutsui K, Jafarzadeh Shirazi MR, et al. The roles of RFamide-related peptides (RFRPs), mammalian gonadotropin-inhibitory hormone (GnIH) orthologues in female reproduction. Iran J Basic Med Sci 2018; 21: 1210-1220. ##10. Mohapatra SS, Mukherjee J, Banerjee D, Das PK, Ghosh PR, Das K. RFamide peptides, the novel regulators of mammalian HPG axis: A review. Vet World 2021; 14: 1867-1873. ##11. Han X, He Y, Zeng G, Wang Y, Sun W, Liu J, et al. Intracerebroventricular injection of RFRP-3 delays puberty onset and stimulates growth hormone secretion in female rats. Reprod Biol Endo-crinol 2017; 15: 35. ##12. Yu C-H, Liu P-H, Van Y-H, Lien AS-Y, Huang T-P, Yen H-R. Traditional Chinese medicine for idiopathic precocious puberty: A hospital-based retrospective observational study. Complement Ther Med 2014; 22: 258-265. ##13. Sun W, Han X, Wang Y, Yu J, Yan W, Zhao J, et al. Effectiveness of Ziyin Xiehuo granules and Zishen Qinggan granules on partial precocious puberty in girls: A multicenter, randomized, sin-gle-blind, controlled trial. J Tradit Chin Med 2018; 38: 740-745. ##14. Huang R, Zeng G-Z, Huang H-Y, Wang Y-H, Yu J. [Further exploration on the essence of yin deficiency caused fire hyperactivity syndrome in precocious puberty children patients]. Zhongguo Zhong Xi Yi Jie He Za Zhi 2016; 36: 879-881. (In Chinese)##15. Zeng G, Han X, Yu J, Wang Y, Tian Z. Effect of nourishing "yin" removing "fire" chinese herbal mixture on hypothalamic mammalian target of rapamycin expression during onset of puberty in female rats. Evid Based Complement Alternat Med 2015; 2015: 157846. ##16. He Y, Han X, Sun W, Yu J, Tamadon A. Precocious puberty and the Lin28/Let7 pathway: The therapeutic effect of the nourishing "Yin" and purging "Fire" traditional chinese medicine mixture in a rat model. Evid Based Complement Alternat Med 2018; 2018: 4868045.##17. Wang S, Zhu L, Yu J, Tian Z. Effect of nourishing “Yin”-removing “Fire” Chinese herbal mix-ture on hypothalamic NKB/NK3R expression in female precocious rats. Evid Based Complement Alternat Med 2014; 2014: 217424. ##18. Bai G-L, Hu K-L, Huan Y, Wang X, Lei L, Zhang M, et al. The traditional chinese medicine fuyou formula alleviates precocious puberty by inhibiting GPR54/GnRH in the hypothalamus. Front Pharmacol 2021; 11: 596525. ##19. Auta T, Hassan AT. 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Chronic demyelination interferes with normal spermatogenesis in cuprizone-intoxicant C57/BL 6 mice: An experimental study دمیلیناسیون مزمن در روند اسپرماتوژنز طبیعی موش ای C57/BL 6 سمی شده با کوپریزون تداخل ایجاد می‌کند: یک مطالعه تجربی 2 1 مقدمه: به دلیل آسیبهای وارده به میلین و آکسون سلولهای عصبی در افراد مبتلا به مولتیپل اسکلروزیس، ممکن است عدم هماهنگی حرکتی و عدم تعادل در ترشحات غدد درونریز ایجاد گردد. هدف: هدف این مطالعه، بررسی نقش دمیلیناسیون مزمن بر محور هیپوتالاموس-هیپوفیز-گناد در مدل موشی مولتیپل اسکلروزیس بوده است. مواد و روش ها: 20 رأس موش C57/BL 6 بالغ نر به دو گروه (10 راس در هر گروه) به شرح ذیل تقسیم شدند: گروه کنترل که یک رژیم غذایی معمول را به مدت 17 هفته دریافت کردند (CONT)، گروه آزمایش (cuprizone [CPZ]) که ابتدا به مدت 12 هفته با غذای حاوی 2/0% سم کوپریزون تغذیه شدند ولی سپس به مدت 5 هفته سم کوپریزون از رژیم غذایی قطع گردید. سطح تستوسترون سرم، هیستوپاتولوژی بافتهای مغز و بیضه، و همچنین پارامترهای اسپرم مورد ارزیابی قرار گرفت. نتایج: محتوای میلین هسته قوسی هیپوتالاموس به طور قابل توجهی پس از 12 هفته مصرف سم کوپریزون در مقایسه با گروه CONT کاهش یافت و این تفاوت آماری تا 17 هفته باقی ماند. سطح تستوسترون به طور قابل توجهی در گروه CPZ در مقایسه با گروه CONT در هفتههای 12 و 17 کاهش یافت. کاهش قابل توجهی در ارتفاع اپیتلیوم لولههای منیساز و مساحت بافت بینابینی و همچنین تعداد انواع سلولهای اپیتلیال لولههای منیساز در گروه CPZ نسبت به گروه CONT، در هفته 12 و 17 مشاهده شد. تعداد اسپرم، تحرک و زندهمانی در گروه CPZ در مقایسه با گروه CONT در هفته 12 و 17 مطالعه به طور قابل توجهی کاهش یافت. نتیجه گیری: دمیلیناسیون مزمن القا شده با سم کوپریزون، ممکن است از طریق آسیبرساندن به هسته قوسی هیپوتالاموس، منجر به اختلال در محور هیپوتالاموس- هیپوفیز- گناد و آسیب به بافت بیضه و متعاقباً تداخل در روند اسپرم‌زایی گردد. 2 Background: Due to myelin and axonal insults in multiple sclerosis individuals, motor coordination problems and endocrine imbalance may develop. Objective: This study aims to evaluate the role of chronic demyelination on the hypothalamic-pituitary-gonadal axis in the mouse model of multiple sclerosis. Materials and Methods: 20 adult C57/BL6 male mice were divided into 2 groups (n = 10/each) as follows: the control group (CONT) received a regular diet for 17 wk; and the experimental group (cuprizone [CPZ]) was fed with 0.2% CPZ for 12 wk and, then CPZ was withdrawn for 5 wk. Serum testosterone, histopathology of the brain and testis, and sperm analysis were evaluated. Results: The hypothalamic myelin content was significantly decreased in the arcuate nucleus following the 12 wk of CPZ consumption compared to the CONT group, and the statistical difference remained until 17 wk. Testosterone levels declined significantly in the CPZ group compared to the CONT group in the 12th and 17th wk. A significant decrease was observed in the height of the seminiferous epithelium and the interstitial tissue area, and the number of seminiferous epithelial cells in the CPZ group compared to the CONT group in the 12th and 17th wk. The sperm count, motility, and viability in the CPZ group significantly decreased compared to the CONT group in the 12th and 17th wk of the study. Conclusion: Chronic demyelination induced by CPZ intoxication, maybe through damage to the hypothalamus arcuate nucleus, leads to the hypothalamic-pituitary-gonadal axis disturbance and damage to the testis and spermatogenesis subsequently. 43 54 2023/06/262023/10/242023/10/262023/07/22023/08/20 1402/5/29 2023/12/252023/12/192024/01/82024/01/132023/12/25 1402/10/4 Arezoo Dorikhani Arezoo Dorikhani Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران arezoo.d.73@gmail.com Ameneh Omidi Ameneh Omidi Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران a.omidi@modares.ac.ir Mansoureh Movahedin Mansoureh Movahedin Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران movahed.m@modares.ac.ir Iman Halvaei Iman Halvaei Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. ایران ihalvaei@modares.ac.ir Multiple sclerosis Cuprizone Hypothalamic-pituitary-gonadal axis Spermatogenesis. مولتیپل اسکلروزیس کوپریزون محور هیپوتالاموس- هیپوفیز- گناد اسپرماتوژنز. 1. Mey GM, Mahajan KR, DeSilva TM. Neurodegeneration in multiple sclerosis. WIREs Mech Dis 2023; 15: e1583.##2. Fymat AL. Multiple sclerosis: I. Symptomatology and etiology. J Neurol Psychol Res 2023; 4: 1.##3. Silveira C, Guedes R, Maia D, Curral R, Coelho R. Neuropsychiatric symptoms of multiple sclerosis: State of the art. Psychiatry Invest 2019; 16: 877-888.##4. Guo Zh-N, He S-Y, Zhang H-L, Wu J, Yang Y. Multiple sclerosis and sexual dysfunction. Asian J Androl 2012; 14: 530.##5. Cavalla P, Rovei V, Masera S, Vercellino M, Massobrio M, Mutani R, et al. Fertility in patients with multiple sclerosis: Current knowledge and future perspectives. Neurol Sci 2006; 27: 231-239.##6. Matuszewska A, Kowalski K, Jawień P, Tomkalski T, Gaweł-Dąbrowska D, Merwid-Ląd A, et al. The hypothalamic-pituitary-gonadal axis in men with schizophrenia. Int J Mol Sci 2023; 24: 6492.##7. Silveira MA, Zampieri TT, Furigo IC, Abdulkader F, Donato Jr J, Frazão R. Acute effects of somatomammotropin hormones on neuronal components of the hypothalamic-pituitary-gonadal axis. Brain Res 2019; 1714: 210-217.##8. Trofimenko V, Hotaling JM. Fertility treatment in spinal cord injury and other neurologic disease. Transl Androl Urol 2016; 5: 102-116.##9. Scandurra C, Rosa L, Carotenuto A, Moccia M, Arena S, Ianniello A, et al. Sexual dysfunction in people with multiple sclerosis: The role of disease severity, illness perception, and depression. J Clin Med 2023; 12: 2215.##10. Racosta JM, Kimpinski K, Morrow SA, Kremenchutzky M. Autonomic dysfunction in multiple sclerosis. Auton Neurosci 2015; 193: 1-6.##11. Milosevic A, Janjic MM, Lavrnja I, Savic D, Bozic ID, Tesovic K, et al. The sex-specific patterns of changes in hypothalamic-pituitary-gonadal axis during experimental autoimmune encephalomyelitis. Brain, Behav Immun 2020; 89: 233-244.##12. Safarinejad MR. Evaluation of endocrine profile, hypothalamic-pituitary-testis axis and semen quality in multiple sclerosis. J Neuroendocrinol 2008; 20: 1368-1375.##13. Labunets I, Rodnichenko A, Savosko S, Pivneva T. Reaction of different cell types of the brain on neurotoxin cuprizone and hormone melatonin treatment in young and aging mice. Front Cell Neurosci 2023; 17: 1131130.##14. Ghaiad HR, Abd-Elmawla M, Gad ES, Ahmed K, Abdelmonem M. Modulating miR-146a expression by hydrogen sulfide ameliorates motor dysfunction and axonal demyelination in cuprizone-induced multiple sclerosis. ACS Chem Neurosci 2023; 14: 3047-3058.##15. Gharighnia S, Omidi A, Ragerdi Kashani I, Sepand MR, Pour Beiranvand Sh. Ameliorative effects of acetyl-L-carnitine on corpus callosum and functional recovery in demyelinated mouse model. Int J Neurosci 2022; 24: 16093.##16. Paxinos G, Franklin KBJ. Paxinos and Franklin's the mouse brain in stereotaxic coordinates. 5th Ed. Cambride: Academic press; 2019.##17. Ziamajidi N, Khajvand-Abedini M, Daei S, Abbasalipourkabir R, Nourian A. Ameliorative effects of vitamins A, C, and E on sperm parameters, testis histopathology, and oxidative stress status in zinc oxide nanoparticle-treated rats. BioMed Res Int 2023; 2023: 4371611.##18. Fletcher SG, Castro-Borrero W, Remington G, Treadaway K, Lemack GE, Frohman EM. Sexual dysfunction in patients with multiple sclerosis: A multidisciplinary approach to evaluation and management. Nat Clin Pract Urol 2009; 6: 96-107.##19. Delaney KE, Donovan J. Multiple sclerosis and sexual dysfunction: A need for further education and interdisciplinary care. NeuroRehabilitation 2017; 41: 317-329.##20. Griswold MD. Spermatogenesis: The commitment to meiosis. Physiol Rev 2016; 96: 1-17.##21. Tomassini V, Onesti E, Mainero C, Giugni E, Paolillo A, Salvetti M, et al. Sex hormones modulate brain damage in multiple sclerosis: MRI evidence. J Neurol Neurosurg Psychiatry 2005; 76: 272-275.##22. Genç B, Şen S, Aslan K, İncesu L. Volumetric changes in hypothalamic subunits in patients with relapsing remitting multiple sclerosis. Neuroradiology 2023; 65: 899-905.##23. Plant TM. The neurobiological mechanism underlying hypothalamic GnRH pulse generation: The role of kisspeptin neurons in the arcuate nucleus. F1000Res 2019; 8: 982.##24. Toolee H, Rastegar T, Solhjoo S, Mortezaee K, Mohammadipour M, Kashani IR, et al. Roles for kisspeptin in proliferation and differentiation of spermatogonial cells isolated from mice offspring when the cells are cocultured with somatic cells. J Cell Biochem 2019; 120: 5042-5054.##25. Qiu W, Raven S, Wu J-Sh, Bundell Ch, Hollingsworth P, Carroll WM, et al. Hypothalamic lesions in multiple sclerosis. J Neurol Neurosurg Psychiatry 2011; 82: 819-822.##26. Krementsov DN, Katchy A, Case LK, Carr FE, Davis B, Williams C, et al. Studies in experimental autoimmune encephalomyelitis do not support developmental bisphenol A exposure as an environmental factor in increasing multiple sclerosis risk. Toxicol Sci 2013; 135: 91-102.##27. Glazer CH, Tøttenborg SS, Giwercman A, Bräuner EV, Eisenberg ML, Vassard D, et al. Male factor infertility and risk of multiple sclerosis: A register-based cohort study. Mult Scler 2018; 24: 1835-1842.##28. Shaygannejad V, Mirmosayyeb O, Vaheb S, Nehzat N, Ghajarzadeh M. The prevalence of sexual dysfunction and erectile dysfunction in men with multiple sclerosis: A systematic review and meta-analysis. Neurología 2022: S2173-5808(22)00088-8.##29. Devis M, Stoquart G, Géonet M, London F, van Pesch V. Validation of the French version of the multiple sclerosis intimacy and sexuality questionnaire-19: A preliminary study. Mult Scler Relat Disord 2022; 65: 103986.##30. Massarotti C, Sbragia E, Gazzo I, Stigliani S, Inglese M, Anserini P. Effect of multiple sclerosis and its treatments on male fertility: Cues for future research. J Clin Med 2021; 10: 5401.## ##

Evaluation of diagnostic value of pelvic MRI in endometriosis in comparison with surgical findings: A cross-sectional study مقایسه دقت تشخیصی MRI لگن در اندومتریوز با یافته های جراحی: یک مطالعه مقطعی 2 1 مقدمه: اندومتریوز یک اختلال ژنیکولوژیک مولتیفوکال طی دوره باروری است. MRI یک روش تشخیصی ارزشمند برای بیماران اندومتریوز است که میتواند هم به تنهایی و هم به صورت همراه با TVS استفاده شود. هدف: این مطالعه با هدف مقایسه دقت تشخیصی MRI لگن در اندومتریوز عمقی با یافتههای جراحی در بیماران مراجعهکننده به بیمارستان شهید صدوقی یزد در مدت یک سال انجام شد. مواد و روش ها: این مطالعه مقطعی بر روی 40 زن مشکوک به اندومتریوز مراجعهکننده به بیمارستان شهید صدوقی یزد از آبان 1399 تا 1400 انجام شد. بیماران بر اساس علائم بالینی، سابقهی جراحی و یا درمان طبی به بخش رادیولوژی جهت مطالعات تکمیلی MRI ارجاع شدند. در نهایت پس از انجام اقدامات تشخیصی لاپاروسکوپیک، یافتههای مشاهده شده در MRI با نتایج پاتولوژیک لاپاروسکوپی مقایسه شد. نتایج: بر اساس یافتههای تصویربرداری MRI و مقایسه آن با یافتههای لاپاروسکوپیک به عنوان استاندارد طلایی تشخیصی، MRI حساسیتی برابر با 8/94%، و اختصاصیتی برابر با 20% در ارتباط با ضایعات مرتبط با اندومتریوز داشت. همچنین ارزش اخباری مثبت (PPV) تصویربرداری MRI برابر با 2/90% و ارزش اخباری منفی (NPV) آن برابر با 3/33% است. نتیجه گیری: با وجود تلاشهای مستمر برای بهبود روشهای تشخیصی و پیشنهادات جدید مانند اضافه کرده سکانس به مدالیتههای تشخیصی MRI، همچنان لاپاروسکوپی به عنوان بهترین و قابل اعتمادترین روش تشخیصی اندومتریوزیز قرار دارد. 2 Background: Endometriosis is a multifocal gynecologic disorder during the fertility period in women. Magnetic resonance imaging (MRI) is an important diagnostic modality for this disease and can be used either alone or along with transvaginal ultrasonography. Objective: This study aims to compare the accuracy of pelvis MRI in pelvic deep endometriosis with laparoscopic findings in women referred to Shahid Sadoughi hospital in one year. Materials and Methods: This cross-sectional study was conducted on 40 women suspicious of endometriosis who referred to Shahid Sadoughi hospital, Yazd, Iran from November 2020-2021. Based on clinical findings and history, participants were referred to the imaging center for pelvic MRI. Finally, the results of MRI and diagnostic laparoscopy were compared with pathologic findings. Results: The sensitivity and specificity of MRI for pelvic endometriosis were 94.8% and 20%, respectively. Also, the positive predictive value and negative predictive value of MRI were 90.2% and 33.3%, respectively. Conclusion: Laparoscopy is still the gold standard of endometriosis diagnosis, but MRI with susceptibility-weighted imaging sequence is the best noninvasive diagnostic method. 55 60 2023/06/262023/10/242023/10/262023/07/22023/08/202022/06/23 1401/4/2 2023/12/252023/12/192024/01/82024/01/132023/12/252023/12/30 1402/10/9 Reza Nafisi Moghadam Reza Nafisi Moghadam Department of Radiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران nafisi.moghadam@gmail.com Fatemeh Tamizi Fatemeh Tamizi Department of Radiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران fatemeh.tamizi@gmail.com Seid Kazem Razavi Ratki Seid Kazem Razavi Ratki Department of Radiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران razavi822@gmail.com Amin Nafisi Moghadam Amin Nafisi Moghadam Faculty of Medicine, Aliebnabitaleb Azad University, Yazd, Iran. ایران aminnafisi1290@yahoo.com Atiyeh Javaheri Atiyeh Javaheri Department of Obstetrics and Gynecology, Shahid Sadoughi Hospital, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران atiyeh56@gmail.com Nasim Namiranian Nasim Namiranian Diabetes Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران namiranian.nasim@gmail.com Deep infiltrating endometriosis Magnetic resonance imaging Diagnosis Laparoscopy Sensitivity Specificity. اندومتریوز نفوذی عمیق MRI تشخیص لاپاراسکوپی حساسیت ویژگی. 1. Guerriero S, Pascual MA, Ajossa S, Rodriguez I, Zajicek M, Rolla M, et al. Learning curve for ultrasonographic diagnosis of deep infiltrating endometriosis using structured offline training program. Ultrasound Obstet Gynecol 2019; 54: 262-269.##2. Taniguchi F, Sakamoto Y, Yabuta Y, Azuma Y, Hirakawa E, Nagira K, et al. Analysis of pregnancy outcome and decline of anti‐Müllerian hormone after laparoscopic cystectomy for ovarian endometriomas. J Obstet Gynaecol Res 2016; 42: 1534-1540.##3. Khodaverdi S, Alebouyeh MR, Sadegi K, Mehdizadehkashi A, Kaveh M, Entezari SR, et al. Superior hypogastric plexus block as an effective treatment method for endometriosis-related chronic pelvic pain: An open-label pilot clinical trial. J Obstet Gynaecol 2021; 41: 966-971.##4. Burla L, Scheiner D, Hötker AM, Meier A, Fink D, Boss A, et al. Structured manual for MRI assessment of deep infiltrating endometriosis using the ENZIAN classification. Arch Gynecol Obstet 2021; 303: 751-757.##5. Culley L, Law C, Hudson N, Denny E, Mitchell H, Baumgarten M, et al. The social and psychological impact of endometriosis on women's lives: A critical narrative review. Hum Reprod Update 2013; 19: 625-639.##6. Nnoaham KE, Hummelshoj L, Webster P, d'Hooghe T, de Cicco Nardone F, de Cicco Nardone C, et al. Impact of endometriosis on quality of life and work productivity: A multicenter study across ten countries. Fertil Steril 2011; 96: 366-373.##7. Di Paola V, Manfredi R, Castelli F, Negrelli R, Mehrabi S, Pozzi Mucelli R. Detection and localization of deep endometriosis by means of MRI and correlation with the ENZIAN score. Eur J Radiol 2015; 84: 568-574.##8. Nisenblat V, Bossuyt PMM, Farquhar C, Johnson N, Hull ML. Imaging modalities for the non-invasive diagnosis of endometriosis. Cochrane Database Syst Rev 2016; 2: Cd009591.##9. Dunselman GAJ, Vermeulen N, Becker C, Calhaz-Jorge C, D'Hooghe T, De Bie B, et al. ESHRE guideline: Management of women with endometriosis. Hum Reprod 2014; 29: 400-412.##10. Burla L, Scheiner D, Pierre Samartzis E, Seidel S, Eberhard M, Fink D, et al. The ENZIAN score as a preoperative MRI-based classification instrument for deep infiltrating endometriosis. Arch Gynecol Obstet 2019; 300: 109-116.##11. Coutinho Jr A, Bittencourt LK, Pires CE, Junqueira F, de Oliveira Lima CMA, Coutinho E, et al. MR imaging in deep pelvic endometriosis: A pictorial essay. Radiographics 2011; 31: 549-567.##12. Bazot M, Lafont C, Rouzier R, Roseau G, Thomassin-Naggara I, Daraï E. Diagnostic accuracy of physical examination, transvaginal sonography, rectal endoscopic sonography, and magnetic resonance imaging to diagnose deep infiltrating endometriosis. Fertil Steril 2009; 92: 1825-1833.##13. Siegelman ES, Oliver ER. MR imaging of endometriosis: Ten imaging pearls. Radiographics 2012; 32: 1675-1691.##14. Krüger K, Behrendt K, Niedobitek-Kreuter G, Koltermann K, Ebert AD. Location-dependent value of pelvic MRI in the preoperative diagnosis of endometriosis. Eur J Obstet Gynecol Reprod Biol 2013; 169: 93-98.##15. Bianek-Bodzak A, Szurowska E, Sawicki S, Liro M. The importance and perspective of magnetic resonance imaging in the evaluation of endometriosis. Biomed Res Int 2013; 2013: 436589.##16. Thomeer MG, Steensma AB, van Santbrink EJ, Willemssen FE, Wielopolski PA, Hunink MG, et al. Can magnetic resonance imaging at 3.0-Tesla reliably detect patients with endometriosis? Initial results. J Obstet Gynaecol Res 2014; 40: 1051-1058.##17. Scardapane A, Lorusso F, Bettocchi S, Moschetta M, Fiume M, Vimercati A, et al. Deep pelvic endometriosis: Accuracy of pelvic MRI completed by MR colonography. Radiol Med 2013; 118: 323-338.##18. Koninckx PR, Ussia A, Adamyan L, Wattiez A, Donnez J. Deep endometriosis: Definition, diagnosis, and treatment. Fertil Steril 2012; 98: 564-571.##19. Berger JP, Rhemrev J, Smeets M, Henneman O, English J, Jansen FW. Limited added value of magnetic resonance imaging after dynamic transvaginal ultrasound for preoperative staging of endometriosis in daily practice: A prospective cohort study. J Ultrasound Med 2019; 38: 989-996.##20. Bielen D, Tomassetti C, Van Schoubroeck D, Vanbeckevoort D, De Wever L, Van den Bosch T, et al. IDEAL study: Magnetic resonance imaging for suspected deep endometriosis assessment prior to laparoscopy is as reliable as radiological imaging as a complement to transvaginal ultrasonography. Ultrasound Obstet Gynecol 2020; 56: 255-266.##21. Stratton P, Winkel C, Premkumar A, Chow C, Wilson J, Hearns-Stokes R, et al. Diagnostic accuracy of laparoscopy, magnetic resonance imaging, and histopathologic examination for the detection of endometriosis. Fertil Steril 2003; 79: 1078-1085.##22. Saba L, Guerriero S, Sulis R, Pilloni M, Ajossa S, Melis G, et al. Learning curve in the detection of ovarian and deep endometriosis by using magnetic resonance: Comparison with surgical results. Eur J Radiol 2011; 79: 237-244.##23. Pin L, Monseau-Thiburce A-C, Ziade-Coularis C, Benjamin A, Menut F, Brun J-L, et al. Exploratory study of the interest of MR susceptibility-weighted imaging for the pre-operative assessment of pelvic endometriosis extent. Eur J Radiol 2019; 118: 245-250.## ##

The genetically modified human foreskin fibroblast cell line (YhFF#8) stably expressing Cas9 gene: A lab resource report دودمانه سلولی تغییر ژنتیکی یافته فیبروبلاست پوست ختنه‌گاه انسان (YhFF#8) با بیان پایدار ژن Cas9: یک گزارش منبع آزمایشگاهی 2 1 مقدمه: ردههای سلولی بیانکننده پایدار Cas9(پروتئین 9 مرتبط با CRISPR) به عنوان ابزار ارزشمندی در تحقیقات ژنتیکی پدیدار شدهاند که کارایی سیستم CRISPR/Cas9 را افزایش داده و روشهای ویرایش ژن را ساده میکنند. این دودمانهای سلولی امکان ویرایش همزمان چندین ژن را فراهم کرده و زمان کلی ویرایش را کاهش میدهد. هدف: این مطالعه با هدف ایجاد یک رده سلولی فیبروبلاست انسانی پایدار با قابلیت تبدیل ژنتیکی به شکل جهش یافته، به عنوان یک مدل سلولی برای یک بیماری ژنتیکی خاص انجام شد. خط سلولی ایجاد شده بررسی مکانیسمهای بیماری، آزمایش درمانهای بالقوه، و به دست آوردن بینش در مورد فرآیندهای مولکولی اساسی را تسهیل میکند. مواد و روش‌ها: سلول‌های 293LTV کلیه جنینی انسان (HEK293LTV) برای تولید ذرات شبهویروس مورد استفاده قرار گرفتند و سلول‌های فیبروبلاست پوست ختنه‌گاه انسان یزد از سلول‌های دسته 8 (YhFF#8) برای تغییرات ژنتیکی هدف قرار گرفتند. ترانزفکشن سلول‌های HEK293LTV با DNA پلاسمیدی نوترکیب pCDH-Cas9 باعث تولید ذرات شبهویروسی شد. سلول‌های YhFF#8 ترانزداکت شده با استفاده از آنتی‌بیوتیک پورومایسین تحت انتخاب قرار گرفتند. درخشش پروتئین فلورسنت سبز (GFP) در سلولهای ترانزداکت شده قبل و بعد از انتخاب آنتیبیوتیکی سلولها تصویربرداری شد. در ضمن سطح بیان نسبی ژن Cas9 در مقایسه با ژنهای GFP و GAPDH با روش کمی (qPCR) تعیین گردید. یافته‌ها: این مطالعه صحت توالی کاست ژن Cas9 و فعالیت رونویسی آن را تأیید کرد. سلول‌های YhFF#8 ترانزداکت شده، فلورسانس سبز نشان داده و با انتخاب آنتی‌بیوتیکی که منجر به خالصسازی تقریباً 100% سلول‌های تبدیل شده شد. بیان ژن GFP با استفاده از میکروسکوپ فلورسانس، امکان رصد زنده سلول‌های YhFF#8-Cas9-GFP-PuroR را در محیط کشت را فراهم کرد. نتیجه‌گیری: سلول‌های YhFF#8-Cas9-GFP-PuroR، برچسب‌گذاری‌شده و مستعد برای انجام فرایند ویرایش ژنومی، منبع بهینه‌ای برای تولید رده‌های سلول‌های بنیادی پرتوان القایی برای تحقیقات زیست‌پزشکی آینده فراهم می‌کنند. 2 Background: Stable Cas9 (CRISPR-associated protein 9)-expressing cell lines have emerged as valuable tools in genetic research, enhancing the efficiency of the CRISPR/Cas9 system and streamlining gene editing procedures. These cell lines enable simultaneous editing of multiple genes and reduce the overall editing time. Objective: This study aimed to develop a stable human fibroblast cell line capable of genetic conversion into a mutant form, serving as a cellular model for a specific genetic disease. The established cell line facilitates investigation of disease mechanisms, testing of potential treatments, and gaining insights into underlying molecular processes. Materials and Methods: Human embryonic kidney 293LTV cells were used to produce pseudo-virus particles, while Yazd human foreskin fibroblasts batch 8 (YhFF#8) cells were targeted for genetic modification. Transfection of human embryonic kidney 293LTV cells with pCDH-Cas9 plasmid DNA generated pseudo-viral particles. YhFF#8 cells were transduced and selected using antibiotics. Green fluorescent protein (GFP) detection confirmed successful transduction and selection. Relative expression levels of the Cas9 gene were determined by quantitative polymerase chain reaction. Results: The study validated the fidelity of the Cas9 gene cassette sequence and its transcriptional activity. Transduced YhFF#8 cells exhibited green fluorescence, with antibiotic selection resulting in nearly 100% transduced cells. A reporter GFP gene enabled real-time monitoring of YhFF#8-Cas9-GFP-PuroR cells using fluorescence microscopy. Conclusion: YhFF#8-Cas9-GFP-PuroR cells, labeled and susceptible to genomic editing, provide an optimal source for generating induced pluripotent stem cell lines for future biomedical research. 61 68 2023/06/262023/10/242023/10/262023/07/22023/08/202022/06/232023/07/24 1402/5/2 2023/12/252023/12/192024/01/82024/01/132023/12/252023/12/302024/01/10 1402/10/20 Farzad Soheilipour Farzad Soheilipour Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Department of Biology, Faculty of Basic Sciences, Gonbad Kavous University, Gonbad Kavous, Iran. ایران soheilipourfarzad@gmail.com Sohrab Boozarpour Sohrab Boozarpour Department of Biology, Faculty of Basic Sciences, Gonbad Kavous University, Gonbad Kavous, Iran. ایران so.boozarpour@gmail.com Shiva Aghaei Shiva Aghaei Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران aghaei.shiva@gmail.com Ehsan Farashahi Yazd Ehsan Farashahi Yazd Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ایران ehsanfarashahi@ssu.ac.ir Fibroblasts Cell line Genetic transduction CRISPR-Cas9. فیبروبلاست ها دودمانه سلولی ترانزداکشن ژنتیکی کریسپر-کاس 9. 1. Carrillo-Ávila JA, Catalina P, Aguilar-Quesada R. Quality control of cell lines using DNA as target. DNA 2022; 2: 44-55.##2. Capes-Davis A, Bairoch A, Barrett T, Burnett EC, Dirks WG, Hall EM, et al. Cell lines as biological models: Practical steps for more reliable research. Chem Res Toxicol 2019; 32: 1733-1736.##3. Dumont J, Euwart D, Mei B, Estes S, Kshirsagar R. Human cell lines for biopharmaceutical manufacturing: History, status, and future perspectives. Crit Rev Biotechnol 2016; 36: 1110-1122.##4. Genzel Y. Designing cell lines for viral vaccine production: Where do we stand? Biotechnol J 2015; 10: 728-740.##5. Peng Y, Ma A, Xiao Zh, Hao J, Feng R, Wang Ch, et al. Technical specifications for ethics review of human stem cell research. Cell Prolif 2023: e13556.##6. Xu Y, Li Zh. CRISPR-Cas systems: Overview, innovations and applications in human disease research and gene therapy. Comput Struct Biotechnol J 2020; 18: 2401-2415.##7. Caneparo Ch, Baratange C, Chabaud S, Bolduc S. Conditioned medium produced by fibroblasts cultured in low oxygen pressure allows the formation of highly structured capillary-like networks in fibrin gels. Sci Rep 2020; 10: 9291.##8. Fernandes IR, Russo FB, Pignatari GC, Evangelinellis MM, Tavolari S, Muotri AR, et al. Fibroblast sources: Where can we get them? Cytotechnology 2016; 68: 223-228.##9. Mesdom P, Colle R, Lebigot E, Trabado S, Deflesselle E, Fève B, et al. Human dermal fibroblast: A promising cellular model to study biological mechanisms of major depression and antidepressant drug response. Curr Neuropharmacol 2020; 18: 301-318.##10. Han X, Liu Z, Jo MC, Zhang K, Li Y, Zeng Z, et al. CRISPR-Cas9 delivery to hard-to-transfect cells via membrane deformation. Sci Adv 2015; 1: e1500454.##11. DeWitt MA, Corn JE, Carroll D. Genome editing via delivery of Cas9 ribonucleoprotein. Methods 2017; 121-122: 9-15.##12. Suchý T, Kaczmarek I, Maricic T, Zieschang C, Schöneberg T, Thor D, et al. Evaluating the feasibility of Cas9 overexpression in 3T3-L1 cells for generation of genetic knock-out adipocyte cell lines. Adipocyte 2021; 10: 631-645.##13. Javaid N, Choi S. CRISPR/Cas system and factors affecting its precision and efficiency. Front Cell Dev Biol 2021; 9: 761709.##14. Aregger M, Chandrashekhar M, Tong AHY, Chan K, Moffat J. Pooled lentiviral CRISPR-Cas9 screens for functional genomics in Mammalian cells. Methods Mol Biol 2018; 1869: 169-188. ##15. Yu H, Wu Zh, Chen X, Ji Q, Tao Sh. CRISPR-CBEI: A designing and analyzing tool kit for cytosine base editor-mediated gene inactivation. MSystems 2020; 5: e00350-20.##16. Dong W, Kantor B. Lentiviral vectors for delivery of gene-editing systems based on CRISPR/Cas: Current state and perspectives. Viruses 2021; 13: 1288.##17. Liao JQ, Zhou G, Zhou Y. Generation of monoclonal iPSC lines with stable Cas9 expression and high Cas9 activity. Methods Mol Biol 2022; 2454: 575-588.##18. Aubrey BJ, Kelly GL, Kueh AJ, Brennan MS, O'Connor L, Milla L, et al. An inducible lentiviral guide RNA platform enables the identification of tumor-essential genes and tumor-promoting mutations in vivo. Cell Rep 2015; 10: 1422-1432.##19. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001; 29: e45.##20. Fernandez JP, Vejnar ChE, Giraldez AJ, Rouet R, Moreno-Mateos MA. Optimized CRISPR-Cpf1 system for genome editing in zebrafish. Methods 2018; 150: 11-18.##21. Lino CA, Harper JC, Carney JP, Timlin JA. Delivering CRISPR: A review of the challenges and approaches. Drug Deliv 2018; 25: 1234-1257.##22. Campa CC, Weisbach NR, Santinha AJ, Incarnato D, Platt RJ. Multiplexed genome engineering by Cas12a and CRISPR arrays encoded on single transcripts. Nat Methods 2019; 16: 887-893. ##23. Lo ChA, Greben AW, Chen BE. Generating stable cell lines with quantifiable protein production using CRISPR/Cas9-mediated knock-in. Biotechniques 2017; 62: 165-174. ##24. Hajizadeh-Tafti F, Golzadeh J, Farashahi-Yazd E, Heidarian-Meimandi H, Aflatoonian B. Established Yazd human foreskin fibroblast lines (#8, #17, and #18) displaying similar characteristics to mesenchymal stromal cells: A lab resources report. Int J Reprod BioMed 2022; 20: 519-528.##25. Ryø LB, Thomsen EA, Mikkelsen JG. Production and validation of lentiviral vectors for CRISPR/Cas9 delivery. Methods Mol Biol 2019; 1961: 93-109.##26. Wang X, Xu Zh, Tian Zh, Zhang X, Xu D, Li Q, et al. The EF-1α promoter maintains high-level transgene expression from episomal vectors in transfected CHO-K1 cells. J Cell Mol Med 2017; 21: 3044-3054.##27. Wen J, Wu J, Cao T, Zhi S, Chen Y, Aagaard L, et al. Methylation silencing and reactivation of exogenous genes in lentivirus-mediated transgenic mice. Transgenic Res 2021; 30: 63-76. ##28. Ellis J. Silencing and variegation of gammaretrovirus and lentivirus vectors. Hum Gene Ther 2005; 16: 1241-1246.##29. 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Aspirin administration from early pregnancy versus initiation after 11 weeks of gestation for prevention of pre-eclampsia in high-risk pregnant women: Study protocol for randomized controlled trial مقایسه ی تجویز آسپرین از اوایل باداری نسبت به شروع مصرف آن از بعد از 11 هفته در پیشگیری از پره اکلامپسی در زنان باردار پرخطر: پروتکل یک مطالعه کارآزمایی بالینی 2 1 مقدمه: پرهاکلامپسی یک اختلال چند ارگانی است که 5-2% زنان باردار تحت تأثیر قرا میدهد. توصیههای ارایه شده برای شروع آسپرین در زنان پرخطر پس از هفته 11 بارداری است. هدف: ما پروتکلی را برای بررسی اثربخشی مصرف آسپرین از اوایل بارداری ارایه میدهیم که یک کارآزمایی تصادفی کنترل شده برای ارزیابی اینکه آیا آسپرین با دوز کم تجویز شده از اوایل بارداری شیوع پرهاکلامپسی زودرس و دیررس را کاهش میدهد. علاوه بر این برای مقایسهی اثربخشی تجویز آسپرین قبل و بعد از 11 هفته در کاهش وقوع پرهاکلامپسی؟ مواد و روش ها: کلیه حاملگیهای در معرض خطر پرهاکلامپسی با توجه به سابقه جمعیتشناختی و مامایی که به کلینیک مادر و جنین بیمارستان دانشگاه تهران مراجعه میکنند برای شرکت در این کارآزمایی دعوت میشوند. نتایج (پیامدهای) بارداری و نوزادان جمعآوری و تجزیه و تحلیل خواهد شد. اولین ثبت نام برای مطالعه پایلوت از شرکتکنندگان پرخطر برای پرهاکلامپسی از ژانویه 2023 بوده است. علاوه بر این ثبت نام در کارآزمایی اصلی از اکتبر 2023 آغاز خواهد شد. 2 Background: Pre-eclampsia (PE) is a multiorgan disorder that affects 2-5% of all pregnant women. Present recommendations for when to start aspirin in high-risk women are after 11 wk of gestation. Objective: We present a protocol to investigate the effectiveness of aspirin use from early pregnancy, which is a randomized controlled trial to assess whether prescribed low-dose aspirin from early pregnancy reduces the prevalence of early and late-onset PE. Additionally, to compare the effectiveness of aspirin administration before and after 11 wk in reducing the occurrence of PE? Materials and Methods: All pregnancies at risk of PE, according to demographic and midwifery history, who are referred to the Maternal-Fetal Clinic of Tehran University hospital, Tehran, Iran were invited to take part in the trial. The outcomes of pregnancy and newborns will be gathered and analyzed. The first registration for the pilot study was in January 2023, and the participants were recognized as high-risk for PE. In addition, enrollment in the main study will begin as of October 2023. 69 80 2023/06/262023/10/242023/10/262023/07/22023/08/202022/06/232023/07/242023/09/12 1402/6/21 2023/12/252023/12/192024/01/82024/01/132023/12/252023/12/302024/01/102023/12/25 1402/10/4 Sedigheh Hantoushzadeh Sedigheh Hantoushzadeh Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Imam Khomeini Hospital Complex, Tehran University of Medical Sciences, Tehran, Iran. ایران hantoushzadeh@tums.ac.ir Arezoo Behzadian Arezoo Behzadian Department of Perinatology, Vali-E-Asr Hospital, Imam Khomeini Hospital Complex, Tehran, Iran. ایران dr.a.behzadian56@gmail.com Mohammad Mehdi Hasheminejad Mohammad Mehdi Hasheminejad Iran University of Medical Sciences, Tehran, Iran. ایران dr.m.mehdi79@gmail.com Faezeh Hasheminejad Faezeh Hasheminejad Asklepios Psychiatry Lower Saxony GmbH, Asklepios Specialist Clinic, Gottingen, Germany. ایران Faezehasheminjd@gmail.com Alireza Helal Birjandi Alireza Helal Birjandi Hannover Medical School, Clinic for Urology, Hannover, Germany. ایران helalbirjandi.alireza@mh-hannovet.de Mojtaba Akbari Mojtaba Akbari Isfahan Endocrine and Metabolism Research Center, Isfahan University of Medicine Sciences, Isfahan, Iran. ایران akbari.epi@gmail.com Marjan Ghaemi Marjan Ghaemi Vali-E-Asr Hospital, Imam Khomeini Hospital Complex, Iran. ایران marjan_ghaemi@yahoo.com Pre-eclampsia Aspirin Color doppler ultrasonography Pregnancy First trimester Pregnancy-associated plasma protein-A. پره اکلامپسی آسپرین سونوگرافی داپلر رنگی بارداری سه ماهه اول پروتئین پلاسمای مرتبط با بارداری-A. 1. Poon LC, Shennan A, Hyett JA, Kapur A, Hadar E, Divakar H, et al. The International Federation of Gynecology and Obstetrics (FIGO) initiative on pre-eclampsia: A pragmatic guide for first-trimester screening and prevention. Int J Gynaecol Obstet 2019; 145 (Suppl.): 1-33.##2. Brosens I, Pijnenborg R, Vercruysse L, Romero R. The “great obstetrical syndrome” are associated with disorders of deep placentation. Am J Obstet Gynecol 2010; 204: 193-201.##3. Zuo Q, Zou Y, Huang Sh, Wang T, Xu Y, Zhang T, et al. Aspirin reduces sFlt-1-mediated apoptosis of trophoblast cells in preeclampsia. Mol Hum Reprod 2021; 27: gaaa089.##4. Benigni A, Gregorini G, Frusca T, Chiabrando C, Ballerini S, Valcamonico A, et al. Effect of low-dose aspirin on fetal and maternal generation of thromboxane by platelets in women at risk for ppregnancy-induced hypertension. N Engl J Med 1989; 321: 357-362.##5. Espinoza J, Romero R, Kim YM, Kusanovic JP, Hassan S, Erez O, et al. Normal and abnormal transformation of the spiral arteries during pregnancy. J Perinat Med 2006; 34: 447-458.##6. Rolnik DL, Wright D, Poon LC, O’Gorman N, Syngelaki A, de Paco Matallana C, et al. Aspirin versus placebo in pregnancies at high risk for preterm preeclampsia. N Engl J Med 2017; 377: 613-622.##7. Hoffman MK, Goudar SS, Kodkany BS, Metgud M, Somannavar M, Okitawutshu J, et al. Low-dose aspirin for the prevention of preterm delivery in nulliparous women with a singleton pregnancy (ASPIRIN): A randomised, double-blind, placebo-controlled trial. Lancet 2020; 395: 285-293.##8. Roberge S, Sibai B, McCaw-Binns A, Bujold E. Low-dose aspirin in early gestation for prevention of preeclampsia and small-for-gestational-age neonates: Meta-analysis of large randomized trials. Am J Perinatol 2016; 33: 781-785.##9. Li C, Raikwar NS, Santillan MK, Santillan DA, Thomas CP. Aspirin inhibits expression of sFLT1 from human cytotrophoblasts induced by hypoxia, via cyclo-oxygenase 1. Placenta 2015; 36: 446-453.##10. Panagodage Sh, Yong HEJ, Da Silva Costa F, Borg AJ, Kalionis B, Brennecke ShP, et al. Low-dose acetylsalicylic acid treatment modulates the production of cytokines and improves trophoblast function in an in vitro model of early-onset preeclampsia. Am J Pathol 2016; 186: 3217-3224.##11. Henderson JT, Whitlock EP, O'Connor E, Senger CA, Thompson JH, Rowland MG. Low-dose aspirin for prevention of morbidity and mortality from preeclampsia: A systematic evidence review for the U.S. preventive services task force. Ann Intern Med 2014; 160: 695-703.##12. Ahrens KA, Silver RM, Mumford SL, Sjaarda LA, Perkins NJ, Wactawski-Wende J, et al. Complications and safety of preconception low-dose aspirin among women with prior pregnancy losses. Obstet Gynecol 2016; 127: 689-698.##13. US Preventive Services Task Force, Davidson KW, Barry MJ, Mangione CM, Cabana M, Caughey AB, et al. Aspirin use to prevent preeclampsia and related morbidity and mortality: US preventive services task force recommendation statement. JAMA 2021; 326: 1186-1191.##14. Roberts JM, King TL, Barton JR, Beck S, Bernstein IM, Buck TE, et al. Care plan for individuals at risk for preeclampsia: Shared approach to education, strategies for prevention, surveillance, and follow-up.3. Am J Obstet Gynecol 2023; 229: 193-213.##15. Poon L, Maiz N, Valencia C, Plasencia W, Nicolaides K. First‐trimester maternal serum pregnancy‐associated plasma protein‐A and pre‐eclampsia. Ultrasound Obstet Gynecol 2009; 33: 23-33.##16. Martin AM, Bindra R, Curcio P, Cicero S, Nicolaides KH. Screening for pre‐eclampsia and fetal growth restriction by uterine artery Doppler at 11-14 weeks of gestation. Ultrasound Obstet Gynecol 2001; 18: 583-586.##17. Harrington K, Carpenter R, Goldfrad C, Campbell S. Transvaginal doppler ultrasound of the uteroplacental circulation in the early prediction of pre‐eclampsia and intrauterine growth retardation. Br J Obstet Gynaecol 1997; 104: 674-681.##18. Brown MA, Magee LA, Kenny LC, Karumanchi SA, McCarthy FP, Saito S, et al. Hypertensive disorders of pregnancy: ISSHP classification, diagnosis, and management recommendations for international practice. Hypertension 2018; 72: 24-43.##19. Cotechini T, Komisarenko M, Sperou A, Macdonald-Goodfellow Sh, Adams MA, Graham ChH. Inflammation in rat pregnancy inhibits spiral artery remodeling leading to fetal growth restriction and features of preeclampsia. J Exp Med 2014; 211: 165-179.##20. Lyall F. Priming and remodelling of human placental bed spiral arteries during pregnancy- a review. Placenta 2005; 26: S31-S36.##21. Roberge S, Bujold E, Nicolaides KH. Aspirin for the prevention of preterm and term preeclampsia: Systematic review and metaanalysis. Am J Obstet Gynecol 2018; 218: 287-293.##22. Bujold E, Roberge S, Lacasse Y, Bureau M, Audibert F, Marcoux S, et al. Prevention of preeclampsia and intrauterine growth restriction with aspirin started in early pregnancy: A meta-analysis. Obstet Gynecol 2010; 116: 402-414.##23. Vainio M, Kujansuu E, Koivisto A-M, Maenpaa J. Bilateral notching of uterine arteries at 12-14 weeks of gestation for prediction of hypertensive disorders of pregnancy. Acta Obstet Gynecol Scand 2005; 84: 1062-1067.##24. Sahin S, Ozakpinar OB, Eroglu M, Tulunay A, Ciraci E, Uras F, et al. The impact of platelet functions and inflammatory status on the severity of preeclampsia. J Matern Fetal Neonatal Med 2015; 28: 643-648.## ##