Introduction and Objective:

Lung cancer is one of the most prevalent and fatal malignancies worldwide. The limitations of conventional therapies, including high toxicity and drug resistance, necessitate the development of novel therapeutic approaches. Green-synthesized iron oxide nanoparticles have gained considerable attention due to their biocompatibility and favorable biological properties. This study aimed to synthesize iron oxide nanoparticles using Urtica dioica extract and evaluate their cytotoxic and apoptotic effects on A549 lung cancer cells.

Materials and Methods:

Iron oxide nanoparticles were synthesized via a green synthesis approach using hydroalcoholic nettle extract. Characterization was carried out using UV-Vis spectroscopy, XRD, FTIR, FESEM, DLS, and zeta potential analysis. The biological effects on A549 cells were assessed using MTT assay, reactive oxygen species (ROS) measurement, mitochondrial membrane potential (MMP) assay, and caspase-3 activity evaluation.

Results:

Characterization analyses confirmed the successful formation of crystalline iron oxide nanoparticles with predominantly spherical morphology. The nanoparticles exhibited dose-dependent cytotoxicity, significantly reducing A549 cell viability. However, no significant change was observed in caspase-3 activity, while increased ROS generation and decreased mitochondrial membrane potential indicated activation of apoptosis pathways. Morphological and nuclear alterations consistent with apoptosis were also observed.
Conclusion:

Green-synthesized iron oxide nanoparticles using Urtica dioica extract effectively induce oxidative stress and apoptosis in lung cancer cells. These findings highlight their potential as promising candidates for the development of novel anticancer therapies.

Keywords:

Iron oxide nanoparticles, Green synthesis, Urtica dioica, Lung cancer, A549, Oxidative stress, Apoptosis

  

  Wounds are any damage or disturbance in the natural structure of the skin that can cause the loss of connection and integrity of the body tissue. Due to the importance of the skin as a vital organ and its role in preventing foreign and infectious agents from entering the body, wound management and treatment is a serious and important issue in modern medicine. One of the approaches in the field of wound management is the use of stem cells. Stem cells are used both locally and systemically to treat skin ulcers. Bone marrow-derived mesenchymal stem cells have been studied with the potential for proliferation and differentiation as an important candidate in wound management studies. The thirsty plant is one of the native plants of the west of the country, which has been used by local people to treat skin wounds in the past. This plant has antioxidant, antimicrobial and immune-modifying properties. In this study, the effect of simultaneous administration of mesenchymal stem cells and thirsty plant extract in experimental skin wound healing in large laboratory mice was investigated and the quality of wound healing at macroscopic and microscopic levels will be evaluated.

چكيده

سابقه و هدف: زخم به هرگونه آسيب و يا اختلال در ساختار طبيعي پوست گفته مي شود. مديريت و درمان زخم، يك موضوع جدي و مهم در پزشكي نوين است. يكي از رهيافت‌ها در زمينه‌ي مديريت زخم استفاده از سلول‌هاي بنيادي ميباشد. سلول‌هاي بنيادي هم به صورت موضعي و هم به صورت سيستميك براي درمان زخم‌هاي جلدي مورد استفاده قرار ميگيرند. سلول‌هاي بنيادي مزانشيمي مشتق شده از بند ناف جنين انسان(ژله وارتون) با داشتن پتانسيل تكثير و تمايز به عنوان يك كانديد مهم در مطالعات مربوط به مديريت زخم، بررسي شده‌اند. انتقال سلول‌هاي بنيادي مزانشيمي از طريق تزريق يا تجويز موضعي به طور مستقيم ميتواند باعث ايجاد مرگ سريع سلولي شود. از اين رو راهبردهاي متفاوتي بر اساس استفاده از مواد و داربست‌ها با هدف افزايش چسبندگي، پروليفراسيون و تمايز سلول‌ها به كار گرفته ميشود. يكي از انواع داربست‌هاي استفاده شده در مهندسي بافت، داربست‌هاي نانوفيبر الكتروريسي شده هستند.  اين مطالعه با هدف بررسي كارايي سلول‌هاي بنيادي مزانشيمي ژله وارتون بارگذاري شده بر داربست نانوفيبر ساخته شده از پلي لاكتيك اسيد / صمغ كتيرا و عصاره پوست انار با روش الكتروريسي ، در ترميم زخم تجربي در موش بزرگ آزمايشگاهي با كمك بافت شناسي و ايمونوهيستوشيمي انجام شد.

مواد و روش‌كار: پس از تهيه پودر پوست انار، عصاره هيدروالكلي پوست آن به روش خيساندن آماده شد. داربست مورد نظر در اين مطالعه با كمك دستگاه الكتروريسي مستقر در آزمايشگاه بهداشت مواد غذايي دانشكده دامپزشكي دانشگاه رازي ساخته شد و عصاره پوست انار در تركيب داربست با نسبت 25/1% استفاده شد. جهت شناسايي داربست از آزمون FTIR وميكروسكوپ الكتروني نگاره استفاده شد. بند ناف جنين full term انسان از بيمارستان بيستون به آزمايشگاه كشت سلول منتقل شده و طبق پروتوكل آزمايشگاه مراحل جداسازي، كشت و پاساژ سلولي انجام شد. جهت سنجش سميت سلولي داربست از آزمون MTT استفاده شد. تعداد 20 سر موش آزمايشگاهي بزرگ نر نژاد ويستار با ميانگين سني دو ماه خريداري و در قفس مخصوص با شرايط محيطي دما 3±22 درجه سانتي‌گراد و سيكل روشنايي(12:12) در خانه حيوانات نگهداري شد و در طول مطالعه، به آب و غذا دسترسي آزاد داشتند. پس از تطابق با شرايط محيطي، موش‌ها با كمك كتامين/زايلازين (ketamine 80 mg/kg & xylazine 10mg/kg) بيهوش شدند. موي سطح پشتي هر موش در محل ايجاد زخم‌ها زده و ضد عفوني شد. در سطح پشتي هر موش، زخم دايرهاي به قطر 20 ميليمتر و با ضخامت كامل ايجاد شد و به طور تصادفي به پنج گروه تقسيم شدند. 1)گروه كنترل منفي 2) گروه تيمارشده با 10⁵ سلول 3) گروه تيمارشده با داربست پلي‌لاكتيك‌اسيد/صمغ كتيرا 4) گروه تيمار شده با داربست پلي‌لاكتيك اسيد/صمغ كتيرا/عصاره انار 5) گروه تيمار شده با داربست پلي‌لاكتيك‌اسيد/كتيرا/عصاره انار بارگذاري شده با 10⁵ سلول . طول مدت تيمار 14 روز بود.. درروزهاي 4، 7، 10 و 14 با كمك دوربين ديجيتال از محل زخم تصاويري اخذ شد و روند ترميم زخم از نظر ماكروسكوپي بررسي شد. پس از 2 هفته، موش‌ها آسان كشي شدند و نمونه‌هاي پوست از ناحيه مورد بررسي اخذ شدند و در محلول فيكساتيو قرار داده شدند. بعد از فيكس شدن بافت ، به روش معمول، مقاطع بافتي از پوست تهيه شد و پس از رنگ آميزي عمومي و اختصاصي با ميكروسكوپ نوري مطالعه و مقايسه شد. همچنين جهت مطالعه ايمونوهيستوشيميايي نمونه‌‌هاي بافتي به آزمايشگاه مربوط ارسال شدند و پروتئينهاي ويمنتين، CD31وki-67 مورد رنگ آميزي و مطالعه قرارگرفتند.

يافته‌ها: ماهيت سلول‌هاي بنيادي مزانشيمي با نتايج فلوسايتومتري و تمايز به رده سلول‌هاي استخواني و چربي تاييد شدند. ماهيت داربست الكتروريسي شده با آزمون FTIR و تصاوير حاصل از ميكروسكوپ الكتروني روبشي تاييد شد. نتايج آزمون MTT هيچ سميت سلولي‌اي را براي داربست نشان نداد. چسبندگي و تكثير سلول‌ها به داربست نيز با استفاده از تصاوير حاصل از ميكروسكوپ الكتروني تاييد شد.   بررسي روند ترميم زخم به صورت ماكروسكوپيك نشان از بهبود موثر در گروه تيمارشده با داربست حاوي عصاره به همراه سلول نسبت به ساير گروه‌هاي درماني بود. مطالع? هيستوپاتولوژيك و هيستومورفومتريك نمونه‌هاي اخذ شده با رنگ‌آميزي هماتوكسيلين-ائوزين و ماسون‌تري‌كروم نشان داد كه گروه‌ درمان شده با داربست حاوي عصاره به همراه سلول‌ ترميم بهتري در زمينه سنتز مجدد بافت پوششي؛ بافت جوانه گوشتي؛ تجمع كلاژن و   رگزايي مجدد را نشان داده و همچنين ميزان التهاب كمتري در مقايسه با ساير گروه‌هاي درماني و گروه كنترل حاصل شد. در بررسي ايمونوهيستوشيمي و شمارش سلولي، كاهش بيان پروتئين ويمنتين در سيتوپلاسم سلول‌هاي فيبروبلاست لايه درم، كاهش بيان ماركر ki-67   در هسته سلول‌هاي لايه بازال اپيدرم و كاهش بيان ماركر CD31 در غشاي سلول‌هاي اندوتليال عروقي ناحيه درم مشاهده شد.

كارايي ضعيف توليد مثلي در گاوهاي شيري همچنان يك نگراني عمده براي صنعت دام شيري در سراسر جهان است. در چند دهه‌ي اخير، انتخاب ژنتيكي براي توليد شير با كاهش كارايي توليد مثلي همراه بوده است. تلاش‌هاي تحقيقي زيادي به منظور ابداع فناوري‌هايي جهت القاءِ تخمك‌گذاري همزمان براي تلقيح در زمان معين (TAI) در گاوهاي گوشتي و شيري انجام شده است. پروتكل Ovsynch، كه شامل دو تجويز هورمون آزادكننده­ي گنادوتروپين (GnRH) به فاصله­ي 9 روز، تجويز پروستاگلاندين F_2? (PGF_2?) هفت روز پس از GnRH اول، و انجام تلقيح 18-16 ساعت پس از تجويز GnRH دوم (GnRH2) است، برنامه­هاي توليد مثلي را مؤثرتر ساخته است. با اين حال، نرخ ضعيف تخمك­گذاري در پاسخ به GnRH2 ممكن است منجر به نرخ­هاي آبستني پايين شود. پژوهش­هاي زيادي جهت بهبود نرخ تخمك­گذاري با جايگزين كردن GnRH2 از جمله با گنادوتروپين كوريونيك انساني (hCG) كه مؤثرتر از GnRH در تحريك تخمك‌گذاري در گاوهاي شيري است انجام شده است. با اين حال گزارش شده است كه اين جايگزيني نرخ‌هاي تخمك‌گذاري و آبستني را افزايش نداد، بنابراين hCG يك جايگزين مناسب براي GnRH2 نيست. ما فرض كرديم كه درصد گاوهايي كه در پاسخ به GnRH2 تخمك­گذاري مي­كنند با تجويز همزمان hCG افزايش مي­يابد. بنابراين در مطالعه­ي حاضر اثر تجويز همزمان hCG و GnRH2 در مقايسه با تجويز جداگانه­ي هر يك از آنها بر عملكرد توليد مثلي گاوهاي هلشتاين شيرده مورد بررسي قرار گرفت. در اين مطالعه 62 رأس گاو بين زايش­هاي دوم و پنجم كه در روزهاي 5 ± 50 پس از زايش خود بودند به­طور تصادفي به سه گروه GPG (Ovsynch)، GPH (مانند گروه GPG ولي تجويز hCG به­جاي GnRH2) و GPG-H (مانند گروه GPG ولي تجويز hCG همزمان با GnRH2) تقسيم و 18-16 ساعت بعد از آخرين تزريق تلقيح (TAI) شدند. دام­ها در روزهاي 1- (TAI = D 0) و 7 جهت تعيين نرخ تخمك­گذاري و در روزهاي 30 و 55 جهت تعيين نرخ­هاي گيرايي و آبستني به روش سونوگرافي معاينه شدند. نمونه­هاي خون از وريد وداج دام­ها در روزهاي صفر و 12 جهت سنجش غلظت­هاي پروژسترون خون اخذ گرديد.

  ikooo, [22.04.25 10:31] Intestinal malignancy presents a worldwide health concern. 1 A significant hurdle in therapeutic intervention involves neoplastic resistance to pharmacological agents, a phenomenon often linked to imbalances within the redox regulatory framework. Zinc oxide nanostructures have demonstrated antineoplastic properties by perturbing this equilibrium. Biocompatible fabrication techniques have positioned ZnO    as a potentially advantageous strategy. The current investigation explores the influence of zinc oxide nanoparticles, generated via both conventional chemical routes and environmentally friendly biogenic pathways, on crucial markers of cellular oxidation and the antioxidant defense network in HT-29 colorectal adenocarcinoma cells. These markers encompass malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH). The central objective is to contrast the impact of these nanostructures on the fluctuations of these indicators and to elucidate the contribution of these alterations to the initiation of programmed cell death within HT-29 cells. In this study, zinc oxide nanoparticles were produced using a sustainable biogenic approach employing pomegranate fruit rind extract. The active phytochemical constituents of the plant-derived material were characterized through gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). The resulting nanostructures underwent comprehensive analysis utilizing X-ray diffraction (XRD), dynamic light scattering (DLS), ultraviolet-visible spectroscopy (UV-Vis), Fourier-transform infrared spectroscopy (FTIR), zeta potential measurements, and field-emission scanning electron microscopy (FE-SEM). 2 The findings substantiated the formation of nanostructures exhibiting a hexagonal wurtzite crystalline arrangement, nanoscale dimensions, and both spherical and hexagonal shapes. 3 Biogenically fabricated zinc oxide nanoparticles were evaluated in parallel with chemically synthesized counterparts on HT-29 cells. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay results indicated that the biogenic nanoparticles displayed more pronounced cytotoxic effects, with an inhibitory concentration 50% (IC50) value of 80.28 ?g/mL. Quantification of oxidative stress biomarkers and antioxidant molecules revealed an elevation in MDA levels coupled with a reduction in SOD and GSH concentrations in the group treated with the biogenic nanoparticles. Acridine orange/propidium iodide (AO/PI) and 4?,6-diamidino-2-phenylindole (DAPI) staining assays corroborated the induction of apoptosis in malignant cells exposed to the biogenic nanoparticles. Zinc oxide nanoparticles synthesized through an eco-friendly method utilizing pomegranate peel extract exhibit considerable promise in enhancing the treatment of colorectal cancer by disrupting the oxidant/antioxidant balance within cancerous cells

  

اين مطالعه به منظور درك هرچه بهتر ارتباط ميان يافته هاي باليني ، هيستوپاتولوژيك و بيان پروتئينهاي مرتبط با آپوپتوز(BCl-2,P53) در اندومتر سگهاي ارجاعي به كلينيكهاي سطح شهر كرمانشاه براي جراحي اوريوهيستركتومي طراحي شده است. مجموع 25 رحم خارج شده به روش جراحي انتخابي، از سگهاي ماده بين سنين 2 تا 12 سال، ابتدا مورد بررسي باليني و هيستوپاتولوژي از جهت وجود هيدرومتر، موكومتر، پايومتر و هيپرپلازي كيستي اندومترقرار گرفته و سپس ماركرهاي BCl-2و P53 با تكنيك ايمونوهيستوشيمي در بافت رحم لوكاليزه و رديابي ميشوند. از حيوانات انتخاب شده قبل از عمل جراحي نمونه خون گرفته شده و ميزان هورمونهاي استراديول ، پروژسترون و فاكتورهاي خوني حيوان ارزيابي ميشوند.

مقادير اندازه‌گيري شده در نمونه‌هاي پاتولوژيك همچنين با مقادير اندازه‌گيري شده در نمونه‌هاي سالم از لحاظ باليني، هيستولوژي بافت رحم، ايمونوهيستوشيمي ماركرهاي مذكور و پارامترهاي خوني 5 قلاده سگ كه مورد اواريوهيستركتومي انتخابي قرار مي‌گيرند مقايسه خواهند شد.

  

  .B, [08.03.25 13:59] One of the common dermatological issues is skin wounds and their healing process. The significance of wound healing arises from its high prevalence in both human and veterinary medicine. The present study aimed to evaluate the effects of a bacterial nanocellulose (BNC)/esculin composite on skin wounds in a rat model. A total of 45 male Wistar rats were used and divided into three groups (15 rats per group): control group, bacterial nanocellulose group, and bacterial nanocellulose/esculin group. Bacterial nanocellulose and bacterial nanocellulose/esculin discs were applied to wounds created on the dorsal region of the rats. On days 7, 14, and 21 post-experiment initiation, digital photographs were taken for morphometric evaluation, and tissues were collected to analyze histopathological changes, hydroxyproline content, glycosaminoglycan (GAG) levels, oxidative/antioxidative parameters, and the gene expression levels of interleukin-1? (IL-1?), transforming growth factor-?1 (TGF-?1), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) in the wound area. The results revealed that bacterial nanocellulose/esculin significantly reduced wound area compared to both the control and bacterial nanocellulose groups throughout the study period. Furthermore, esculin treatment reduced lymphocyte count compared to the control group, decreased fibroblast count in the early phases, and increased fibrocyte count in the later stages of wound healing. Other parameters such as re-epithelialization, tissue organization, collagen fiber maturation, and larger-sized blood vessels showed significant improvements in the bacterial nanocellulose/esculin group compared to the other groups. In the bacterial nanocellulose/esculin-treated group, dry matter content, glycosaminoglycan levels, and hydroxyproline content were significantly increased during different wound healing stages compared to the control group. Evaluation of oxidative stress parameters also indicated improved antioxidant status in the wound tissue of the bacterial nanocellulose/esculin-treated group, as evidenced by increased glutathione peroxidase and superoxide dismutase activity and decreased malondialdehyde concentration. Moreover, esculin treatment significantly modulated gene expression patterns at different stages of wound healing, which clinically contributed to accelerated recovery and reduced inflammatory complications. The findings of this study demonstrated that bacterial nanocellulose/esculin is effective in wound healing. The accelerated wound healing properties of this formulation could be attributed to its effects on vascularization, modulation of pro-inflammatory cytokines, enhanced antioxidant defense mechanisms, and increased collagen and glycosaminoglycan production. Keywords: Bacterial Cellulose, Esculin, Skin Wound, Rat.

Colorectal cancer (CRC) is one of the most prevalent and fatal types of cancer. Despite advances in therapeutic strategies, the limitations and side effects of current treatments underscore the urgent need for novel and more effective approaches. Among these, zinc oxide nanoparticles (ZnO   ) have emerged as a promising candidate for cancer therapy due to their significant antitumor properties and the feasibility of environmentally friendly synthesis methods. While numerous studies have explored the antitumor effects of ZnO   , the precise mechanisms of action and their relationship with alterations in tumor marker expressions, such as carcinoembryonic antigen (CEA) and Kirsten rat sarcoma viral oncogene homolog (KRAS), remain inadequately understood.

This study aims to elucidate the dual effects of green-synthesized ZnO    on HT-29 colorectal cancer cells and their impact on the expression of these tumor markers, contributing to the personalization of treatments and improvement of therapeutic outcomes for CRC patients. Additionally, to assess the role of synthesis methods in nanoparticle efficacy, the results of green-synthesized ZnO    were compared with those of chemically synthesized ZnO   .

Green synthesis of ZnO    was performed using ethanolic pomegranate peel extract (Punica granatum). Bioactive compounds in the extract were identified using gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). The synthesized nanoparticles were characterized using X-ray diffraction (XRD), dynamic light scattering (DLS), ultraviolet-visible spectroscopy (UV-Vis), Fourier-transform infrared spectroscopy (FTIR), zeta potential analysis, and field-emission scanning electron microscopy (FE-SEM). The results confirmed the formation of nanoscale particles with a hexagonal wurtzite crystal structure, spherical and hexagonal morphology.

The antitumor effects of the green-synthesized ZnO   , compared to chemically synthesized ZnO   , were evaluated on colorectal cancer cells. The MTT assay revealed that green-synthesized ZnO    exhibited stronger dose-dependent cytotoxic effects on colorectal cancer cells than their chemically synthesized counterparts, with an IC50 value of 80.28 µg/mL. DAPI and acridine orange/ethidium bromide staining demonstrated significant tumor cell growth arrest in cancer cells treated with green-synthesized ZnO   . Furthermore, ELISA analysis showed a substantial reduction in tumor biomarkers CEA and KRAS.

The findings of this study suggest that integrating novel therapeutic approaches, including the application of green-synthesized ZnO    using ethanolic pomegranate peel extract (P.G.), along with precise evaluation of multiple tumor biomarkers, could lead to significant advancements in targeted and effective colorectal cancer therapies.

Keywords: Zinc oxide nanoparticles, green synthesis, colorectal cancer cells, HT-29, tumor biomarkers, ELISA, DAPI staining, acridine orange/ethidium bromide.

Colorectal cancer is one of the most significant health threats, with current treatments facing challenges such as severe side effects and cellular resistance. Inducing apoptosis has emerged as an innovative approach to overcoming these obstacles. Green-synthesized nanoparticles have shown great promise in this regard. These nanoparticles can effectively eliminate cancer cells by inducing apoptosis while minimizing side effects.

In this in vitro study, aimed at evaluating antitumor potential and apoptosis induction, zinc oxide nanoparticles (ZnO   ) were synthesized via a green method using ethanolic pomegranate peel extract (Punica granatum). Initially, bioactive compounds in the pomegranate peel extract were identified using gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). The nanoparticles were subsequently characterized using techniques such as X-ray diffraction (XRD), dynamic light scattering (DLS), ultraviolet-visible spectroscopy (UV-Vis), Fourier-transform infrared spectroscopy (FTIR), and field-emission scanning electron microscopy (FE-SEM). The results confirmed the formation of ZnO nanoparticles with a hexagonal wurtzite crystal structure, nanoscale size, and spherical and hexagonal morphology.

The antitumor effects and apoptosis-inducing capabilities of green-synthesized ZnO    were compared with chemically synthesized ZnO    on colorectal cancer cells (HT-29). MTT assay results demonstrated that green-synthesized ZnO    exhibited stronger dose-dependent cytotoxic effects than their chemically synthesized counterparts. The IC50 value for green-synthesized nanoparticles was determined to be 80.28 µg/mL, indicating higher cytotoxicity compared to chemically synthesized ZnO   . Acridine orange/propidium iodide and DAPI staining techniques revealed a significant increase in apoptosis in cancer cells treated with green-synthesized ZnO   . Furthermore, immunocytochemistry (ICC) analysis showed a significant increase in p53 protein levels and a substantial decrease in Bcl-2 protein expression in these cells (p ? 0.0001).

These findings indicate that green-synthesized ZnO    exert stronger cytotoxic and apoptosis-inducing effects on cancer cells compared to chemically synthesized nanoparticles. The results of this study clearly demonstrate that green synthesis of ZnO    using pomegranate peel extract (P.G.) offers much higher potential for inducing cellular apoptosis and inhibiting tumor cell growth. These findings open new horizons in cancer nanotechnology, suggesting that green-synthesized ZnO    could serve as a promising therapeutic agent in cancer treatment.

Keywords: Apoptosis, BCL-2, p53, colorectal cancer cells HT-29, zinc oxide nanoparticles, green synthesis, immunocytochemistry (ICC).

and Objective: Gills are an organ that is highly

gills included primary filaments that extended parallel from the arch axis and were predominantly covered by marginal epithelial cells with a fingerprint-like surface appearance. Pores of mucous cells and chloride cells were observed among the epithelial cells. Perpendicular to the primary filaments, secondary filaments extended parallel and their surfaces were covered by marginal epithelial cells. At the ends of the gill rakers, taste buds and mucous cells were frequently seen

Conclusion: The gills of longspine Scarper (Paracapoeta trutta. Heckel 1843) are similar to the gills of other bony fish under microscopic examination.  

Keywords: gill, long-finned black fish, histology, surface ultrastructure

Abstract

Introduction and aim: skin is one of the important organs in aquatic animals, especially bony fish and amphibians. Due to its specialized cells and structures, it plays a role not only in the obvious function of protection against environmental factors but also in maintaining and regulating osmotic balance, sensory perception, and the secretion of certain substances. The health status of the skin and its structural changes reflect the health of the fish and are key to understanding the progression of diseases and the impact of environmental changes on fish. On the other hand, fish skin today has applications in other industries such as medicine, pharmaceuticals, human and animal nutrition, and also in cellular and tissue research, leading to an increasing interest in its study. This interest aims not only to reduce costs associated with environmental pollution from fish farming waste but also to utilize these resources in other fields. Therefore, understanding fish skin and its structures is of great importance. In this study, the skin of the Longspine Scarper (Paracapoeta trutta Heckle 1843), as one of the native fish of the western region of the country, will be examined using light microscopy, and the surface ultrastructure of the skin will be studied using scanning electron microscopy.

Material and method: In the present study, five specimens of long-finned black fish were caught from the Sirvan River at the site of the Daryan Dam. After being transferred to the histology laboratory, 1 cm by 1 cm skin samples were taken from the dorsal and ventral surfaces of the anterior and posterior regions and immersed in containers containing fixative solution. After fixation, the samples underwent tissue processing, and tissue sections were prepared. Subsequently, the tissue sections were subjected to general staining and specific staining with Masson's trichrome, periodic acid-Schiff, and Alcian blue, and were examined using a light microscope. Necessary images were captured with a digital camera. Histometric analysis was performed using ImageJ software, and the resulting data were subjected to statistical analysis. Additionally, tissue samples from the dorsal and ventral skin of the fish were taken, and after fixation and dehydration, they were sent to the electron microscopy laboratory for processing and to obtain electron images of their surfaces.

Result: The results of this study showed tha, the skin of the the Longspine Scarper consisted of three structural layers: epidermis, dermis, and hypodermis. The epidermis comprised a living and active stratified squamous epithelium made up of epithelial cells in three forms: cylindrical to cuboidal on the basement membrane, polygonal, and flat squamous at the surface of the epidermis. Additionally, goblet and club cells were observed among the epithelial cells. The dermis was divided into two layers, loose and dense, beneath the basement membrane, extending to broad muscle bundles, and contained blood vessels, pigment cells, and fat cells, which also encompassed the scales. From a histometric perspective, the thickness of the epidermis on the ventral surface in both anterior and posterior regions was greater than that on the dorsal surface, with the thickness in the posterior regions being greater than in the anterior regions. In counting secretory cells, the craniodorsal and caudoventral regions had the highest number of goblet and club cells, showing a significant difference (P<0.05) compared to other studied regions. A significant difference (P<0.05) was observed in the comparison of two cell populations in each region, except for the anterior ventral region in the other three regions. The scales of the black longfin fish were of the cycloid type, and in the ultrastructural study using an electron microscope, the arrangement of scales on the skin surface and their posterior regions covered by the epidermis were observable, along with the patterns of micro-ornamentation of the surface cells and the head pores among them. The structural features of the surface scales included rounded protrusions and transverse grooves, and their arrangement in the anterior, posterior, and lateral regions of each scale was similar to cycloid scales in other related species. However, the eccentricity of the focus area, the convex anterior and posterior edges, and the central and short lateral line canal in the scales along the lateral line were distinctive features of this species' scales

Conclusion: the general structure of the skin tissue of Longspine scarper was similar on both the dorsal and ventral surfaces. However, the skin thickness on the ventral surface was greater than that on the dorsal surface. Additionally, from a histological, histochemical, and ultrastructural perspective, it was similar to other freshwater fish

Keywords: Longspine Sca

  استفاده وسيع از داروهاي شيميدرماني در درمان انواع سرطانها منجر به ايجاد سميت در ارگانهاي مختلف از جمله كليه ميشود. دوكسوروبيسين از جمله اين داروهاست كه سميت كليوي آن به اثبات رسيدهاست. از آنجايي كه سيرينجيك اسيد، يك آنتياكسيدان معروف و از دسته فنوليكاسيدها ميباشد، در اين مطالعه بر آن شديم تا اثرات محافظتي احتمالي سيرينجيك اسيد بر سميت كليوي ناشي از دوكسوروبيسين را مورد بررسي قرار دهيم. در اين مطالعه از 35 سر موش صحرايي ويستار نر استفاده خواهد شد. حيوانات به صورت تصادفي به 5 گروه تقسيم شده و به مدت 14 روز بهصورت ذيل تحت تيمار بهروش خوراكي قرار ميگيرند: كنترل منفي )سرم فيزيولوژي(، كنترل مثبت )سرم فيزيولوژِي(، كنترل مقايسهاي )ويتامين C )و تيمار 1 و 2 )سيرينجيك اسيد با دوزهاي 50 و 100 ميليگرم بر كيلوگرم(. ضمنا تمامي گروهها بهجز كنترل منفي در روز 11م دوكسوروبيسين درونصفاقي دريافت خواهند كرد. پس از اتمام دوره درمان، حيوانات به مدت 24 ساعت پرهيز غذايي دادهشده و روز 15م آسانكشي شده و از كليهآنها جهت آزمونهاي هيستوپاتولوژي، ايمونوهيستوشيمي )سنجش شاخصهاي مربوط به آپوپتوز شامل -3Caspase و 2BCl )و بيوشيميايي )سنجش شاخصهاي استرس اكسيداتيو شامل مالون دي آلدهيد، سوپراكسيد ديسموتاز و ظرفيت آنتي اكسيداني تام( نمونهگيري خواهد شد.

  

The effects of co-administration of Doxorubicin and green synthesized ZnO    on the osteogenic differentiation of bone marrow derived mesenchymal stem cells; in vitro and in silico assessment

Abstract

The chemotherapy drug Doxorubicin (DOXO) can inflict substantial bone damage on cancer patients. While the mechanisms behind DOXO-induced osteoporosis remain incompletely elucidated, evidence suggests that DOXO may hinder the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) through the Bone Morphogenetic Protein-2 (BMP-2)/Drosophila mothers against decapentaplegics (SMAD) signaling pathway. Notably, Zinc Oxide Nanoparticles (ZnO   ) have proven effective in promoting bone formation, mineralization, and osteoblastic cell proliferation. Recently, green-synthesized ZnO    exhibit immense potential for various biomedical applications due to their biocompatibility and biodegradability.

This study investigates the effects of co-administration of DOXO and green synthesized the ZnO    from hydroalcoholic extract of Cercis siliquastrum (C.S) on the osteogenic differentiation of BMSCs; in vitro and in silico assessments. ZnO    were synthesized from hydroalcoholic extract of C.S and characterized both qualitatively and quantitatively. To determine their effect on osteogenic differentiation, BMSCs were cultured in media with and without ZnO    (10 µg/ml) and DOXO (10 nmol) for 14 days. The transcription of genes involved in osteogenic differentiation of BMSCs was analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). The impact of ZnO    on osteoblast function and bone formation was assessed by ALP enzyme and intracellular calcium deposition assays. To investigate the ability of the ZnO    to target BMPRs, an in silico molecular docking simulation was conducted using the descriptive-analytical method. AutoDock 4.2.6 software was employed to examine the binding interaction between the ZnO    and the active site of BMPRs.

The BMSCs groups treated with the ZnO    significantly increased the expression of osteogenic differentiation-related genes (BMP-2, runt-related transcription factor 2 (RUNX2), ALP and collagen type IA (COL1A). and intracellular calcium deposition. Also the most negative binding energy level (?G bind = -3.14 and -2.32 kcal/mol) demonstrated a propensity for binding to the active sites of both the type II receptor (BMPRII) and type I receptor (BMPRIa), Respectively.

Our research sheds valuable light on the mechanism underlying the osteogenic differentiation-inducing effects of a combined treatment of DOXO and the ZnO   , both in vitro and in silico. This finding could potentially pave the way for exploring novel strategies to prevent chemotherapy-induced osteoporosis.

Key words:

Doxorubicin, bone marrow mesenchymal stem cells, zinc oxide nanoparticles, Cercis siliquastrum, molecular docking, BMP2 signaling pathway

The present study was conducted for investigating the sperm morphology and morphometry of Kurdish stallions. The semen samples were collected from 10 Kurdish stallions. Three preparations from each ejaculate were stained with eosinnigrosin (EN), Diff-Quik (DQ), and Rose Bengal (RB). The area, perimeter, length, and width of the sperm head as well as tail length and total sperm length were measured. The parameters ellipticity, elongation, roughness, and regularity were calculated. The morphology of sperm was also investigated under scanning and transmission electron microscopes. DQ and RB provided more clarified images for examining sperm structures compared to the EN method. The head length, head width, area, and perimeter in EN were significantly higher than those in DQ and RB (P?0.05). Furthermore, the difference in head width, head area, and head perimeter between DQ and RB was not significant (P?0.05). The tail length and total sperm length in all methods were close together (P?0.05). The highest percentage of normal sperm was seen in DQ and RB methods (82.55±2.88 and 88.31±5.19) respectively. The highest values for ellipticity, elongation, and 55 regularity were found in RB, whereas the highest value for roughness was measured in EN. Tail defects including coiled tails, and folded midpieces were the most frequent. Scanning electron microscope revealed two types of head shapes: heads with round anterior border, and heads with flat anterior border. The results indicated that despite the routine use of EN for morphological assessment of stallion sperm, RB and DQ can be considered for more clarified details of sperm structure including acrosome and midpiece. Furthermore, the Kurdish stallion sperm has morphometric traits in the normal range established for stallions, however, some traits were larger than those reported for other breeds. It seems that the sperm of the Kurdish stallion has a longer head and tail in comparison to other horse breeds.

The present study was conducted for investigating the sperm morphology and morphometry of Kurdish stallions. The semen samples were collected from 10 Kurdish stallions. Three preparations from each ejaculate were stained with eosin-nigrosin (EN), Diff-Quik (DQ), and Rose Bengal (RB). The area, perimeter, length, and width of the sperm head as well as tail length and total sperm length were measured. The parameters ellipticity, elongation, roughness, and regularity were calculated. The morphology of sperm was also investigated under scanning and transmission electron microscopes. DQ and RB provided more clarified images for examining sperm structures compared to the EN method. The head length, head width, area, and perimeter in EN were significantly higher than those in DQ and RB (P?0.05). Furthermore, the difference in head width, head area, and head perimeter between DQ and RB was not significant (P?0.05). The tail length and total sperm length in all methods were close together (P?0.05). The highest percentage of normal sperm was seen in DQ and RB methods (82.55±2.88 and 88.31±5.19) respectively. The highest values for ellipticity, elongation, and regularity were found in RB, whereas the highest value for roughness was measured in EN. Tail defects including coiled tails, and folded midpieces were the most frequent. Scanning electron microscope revealed two types of head shapes: heads with round anterior border, and heads with flat anterior border. The results indicated that despite the routine use of EN for morphological assessment of stallion sperm, RB and DQ can be considered for more clarified details of sperm structure including acrosome and midpiece. Furthermore, the Kurdish stallion sperm has morphometric traits in the normal range established for stallions, however, some traits were larger than those reported for other breeds. It seems that the sperm of the Kurdish stallion has a longer head and tail in comparison to other horse breeds.

  

  Osteoarthritis is one of the most common skeletal problems that affects millions of people around the world and makes life difficult for these people. There are many methods for inducing osteoarthritis in various researches, which are divided into two general categories: mechanical and chemical. Mechanical method includes surgical methods. including a variety of surgical models, including partial or complete meniscectomy, medial meniscus destabilization, meniscal tear, anterior cruciate ligament (ACL) or posterior cruciate ligament amputation, medial and/or external collateral ligament amputation, cartilage defect, and osteotomy is Among the mechanical methods, cutting the anterior cruciate ligament is one of the common methods for causing osteoarthritis. Mesenchymal stem cells (MSCs) are non-hematopoietic progenitor cells that can differentiate into different tissues such as cartilage, osteocyte and fat cells. Together, stem cells secrete nutritional, vascular, and immunosuppressive factors that have a paracrine effect on tissue resident cells (TRC). Mesenchymal stem cells are usually obtained from bone marrow, but with today's knowledge, these cells can be isolated from different tissues such as skeletal muscle, synovial membrane, and fat tissue. It has recently been found that extracting stem cells from adipose tissue is more suitable due to less invasiveness and risk. In adult animals, adipose tissue accumulates mostly in the area of the arm, thigh, subcutaneous part of the abdomen, omentum, and fat around internal organs such as kidney and liver. The amount of body fat depends on the physical condition (obesity/thinness) of the animal or human, but the amount of omentum has a fixed value. Also, the access to omentum tissue is easier than other tissues and it is more efficient than subcutaneous fat tissue. The purpose of this study is to investigate the effect of mesenchymal cells extracted from the omentum in the treatment of experimental osteoarthritis in a rabbit animal model. For this purpose, 22 pieces of adult male rabbits, with an average weight of 1300 ± 200 grams, will be used, and in all rabbits, after anesthesia, the anterior cruciate ligament of the left knee will be cut. After 12 weeks, two pieces of rabbit will be humanely euthanized and osteoarthritis will be confirmed using radiology and microCT. Other rabbits are divided into 4 groups of 5. The first group is the control group, which does not receive any treatment. The second group will be treated with omentum mesenchymal cells, the third group will be treated with hyaluronic acid, and the fourth group will be combined treatment with mesenchymal cells and hyaluronic acid. 8 weeks after treatment, all animals will be humanely euthanized and subjected to microCT and pathology studies. It is expected that by comparing the treatments that have been carried out, it will provide a suitable treatment method for osteoarthritis.

  Abstract Habronemiasis in Horses is caused by species of Habronema nematode and transmitted by insect carriers, importantly the flies’ species including Muscae domestica and Stomoxys calcitrans. Due to importance of diseases spreading by arthropod carriers, alongside the fact that main horse training centers and farms disregarding this disease and specially its hygienic and economical aspects, this made us up to achieve supporting estimation of ocular Habronemiasis occurrence through certain diagnostic methods for this parasite via molecular tracking and also calculation of Habronema species prevalence at important horse breeding farms and horseracing centers in Kermanshah City. This study was taken place during a 14 month period by observing 41 centers (Horseracing Clubs, Breeding farms and personal stables) and population equal to 385 Horses. By considering clinical history of horses and clinical signs of Ocular Habronemiasis, 28 suspicious horses were detected and grouped in a lower society named “Targets” and they went undergone sampling from lesions of conjunctivae tissue in addition with their environmental disease carrier flies collected as second samples for proving Habronema parasitic presence and investigating Habronema species prevalence through Habronema Genome tracing via Real-time qPCR method. After molecular test procedure and statistically analyzing obtained data, 75% of the suspicious horses were reported Positive for being infected by Habronema, while the positive Targets consists of 61.9% male and 38.1% female. Although the relation of disease occurrence with factors like age of horses, season of disease occurrence, races and body colors was meaningful, indicating about 80.95% of positive targets were from Kurdish race and about 42.85% were Sorrel for their body color. Among the whole 385 horses under observation, horses with age range below 5 years old had the most infection rate and the disease prevalence was at highest peak at summer and after that, at fall. Carrier flies were sampled from suspicious Horses living places, were tested and indicate presence of Habronema Larvae in 92.85% of Targets’ environment. For environments of positive Target horses, all of places had Habronema Larvae detected in flies. As the matter of determination of Habronema species, in molecular investigation of whole positive target samples were reported 92.85% and 7.15% respectively for Habronema muscae and Habronema microstoma. Organized fundamental anti-parasitic treatment programs, sanitizing horsebox floor regularly, preventing from formation of insects attracting places from excrements all over the farm and ultimately medical management of conjunctivitis lesion treatment might be effective in reducing occurrence of ocular Habronemiasis

The objective of this study was to investigate thepattern of changes in vaginal epithelial cells and serum progesterone andestrogen concentrations during the estrous cycle and early pregnancy inmultiparous Synjabi ewes. The ewes (n=20) 45 to 60 days in milk weresynchronized by intramuscular administration of GnRH (day 0)-PGF2? + hCG (day7) and insertion of a controlled internal drug release device (CIDR) (days0-7). At day 7, the ewes were introduced to four fertile rams and observed forestrus behavior. On exhibiting estrus signs, the ewes (n=14) were allocatedrandomly to the study groups: 1) Pregnant (n=9): the ewes were allowed to bemated, then isolated and housed in a separate location. 2) Non-pregnant (n=5):the ewes were not allowed to be mated and isolated from the rams immediatelyafter being detected in estrus. From the first day of exhibiting estrus signs(day 0) until day 20, mucosal samples were collected daily from the anteriorvagina of all ewes using a cytobrush and three smears were prepared from eachsample. The smears were stained using Giemsa staining and studied under lightmicroscopy (Objective: x40) to count cell types. The percentage of each cell typewas calculated as the number of the corresponding cell type counted within 10microscopic fields divided by the total number of all cell types. Blood sampleswere collected via puncture of the jugular veins of all animals intonon-heparinized plastic tubes every other day beginning at day 0 andtransmitted to the laboratory within one hour to determine changes in serumconcentrations of progesterone and estrogen during the first 20 days afterestrus detection. The serum was obtained after centrifugation (3000x g, 15minutes), and stored at -20 ^?C until hormonal assay using ELISA.Pregnancy diagnosis in the Pregnant group was performed 35 days post-mating bytransrectal ultrasound and the pregnant ewes (n=5) served as the Pregnantgroup, while those that were non-pregnant, were excluded from the study. Theresults showed difference in the percentage of vaginal epithelial cells amongvarious stages of estrus cycle and early pregnancy. In estrus and metestrus, nosignificant differences were observed in the percentage of each cell typebetween both groups. In these stages, the greatest and the least percentages ofthe cells were those of the superficial and parabasal cells,respectively. In diestrus, neutrophils and the keratinized cells were greatestin the Pregnant and Non-pregnant ewes, respectively. In this stage, thepercentage of superficial cells showed a remarkable reduction in both thePregnant and Non-pregnant groups. The number of intermediate cells reduced inthe Non-pregnant group but at the same time, they were constant in the Pregnantgroup. The parabasal cells were the least cell population in both groups. Inthe last 4 days of sampling, neutrophils were the greatestcells in the Pregnant group, whereas thesuperficial cells were greatest in the Non-pregnant group. At this stage,neutrophils showed a considerable reduction in the Non-pregnant group, but thenumber of parabasal and intermediate cells were significantly increased. At day0, serum progesterone concentrations were <1 ng/ml in both groups. Then itgradually increased to reach   its maximumconcentration. Maximum progesterone concentrations were maintained in thePregnant group until day 20, but in the Non-pregnant group they started to decrease from day 16 and reached <1ng/ml on days 18 to 20. The highest levels of estrogen were observed on day 0in both groups, then reached <2 pg/ml and remained at this level until theend of the period, whereas in the Non-pregnant group, estrogen concentrationsagain returned to the maximum values on day 18. In conclusion, the results ofthe present study showed that vaginal cytology can be used as a useful tool inassessing hormonal and physiological characteristics of the reproductive systemof ewes and thus provides a more accurate understanding of the physiology ofthe estrus cycle and early pregnancy in ewes, which can be used to improve

reproductive management.  

Introduction:

  

Method:

  

Result and Discussion: