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The purpose of this research was to determine the correlation between the weight of the ovary, the number of follicles, and the quality of the oocytes from the culling female bovine obtained from the slaughterhouse. The variation may affect the potential of the ovary to produce follicles. Paired ovaries were obtained from female bovines. Follicle numbers were recorded and divided into three size categories (small: 3 mm, medium: < 3 mm-8 mm, and large: > 8 mm). Oocytes were aspirated and the number of oocytes was recorded and graded into four categories (grades A, B, C, and D). There was a positive correlation between the weight of the ovary and the number of follicles, which is 0.560 with the regression equation y = 3.52 + 0.501 x. There was a positive correlation between the number of follicles and the number of oocytes, which is 0.546 with the regression equation y =2.48 + 1.204 x. There was a positive correlation between the number of oocytes and the grade of oocytes, which is 0.520, with the regression equation y = 0.93 + 0.800 x. There was no correlation between the weight of the ovary and oocyte grade A, which is 0.013.

Objective: Mitotically active cellular fibroma (MACF) of the ovary, characterized by relatively high mitotic activity without severe atypia, was first described in the WHO classification in 2014. However, due to its rarity, the clinicopathological characteristics of ovarian MACF have not been established. This study was performed to describe the clinical, radiological, and pathological features of MACF by analyzing 11 cases of ovarian MACF. Materials and methods: Between 2015 and 2022, 11 patients with ovarian MACFs underwent surgical treatment at our institution. Clinicopathologic data of the patients were retrospectively reviewed from their medical records. Results: Median patient age was 53.7 years (range 21–77 years), and median tumor diameter was 7.8 cm (range 4.3–14.0 cm). Preoperative CA125 was elevated in 4 cases. Four of the eleven patients had abdominal pain, and two presented with vulvar pain or a palpable abdominal mass, respectively. Preoperative radiological impressions included fibroma, fibrothecoma, stromal tumor, and cystadenocarcinoma. A laparoscopic approach was adopted in 7 cases (64%). Intraoperative frozen section was performed in 5 patients, and all demonstrated the presence of a benign, fibromatous stromal tumor. Three patients underwent fertility-sparing surgery, including laparoscopic ovarian cystectomy and unilateral salpingo-oophorectomy. Median follow-up was 37.7 months (range 2–84 months), and no patient experienced disease relapse or died of their disease. Conclusion: This study shows that ovarian MACF has a benign clinical course. Fertility-sparing surgery provides a safe therapeutic option for MACF, which can be managed safely by laparoscopy. Imaging findings and final pathological diagnosis were not well matched. Intraoperative frozen section is important for determining surgical extent in mitotically active cellular fibroma of the ovary.

Ovarian lymphangiomas are rare benign neoplasms characterized by the proliferation of lymphatic vessels within the ovarian tissue. While lymphangiomas can manifest in various anatomical locations, their occurrence within the ovaries is exceptionally uncommon, posing diagnostic and therapeutic challenges for clinicians. The aetiology of ovarian lymphangiomas remains elusive, with theories suggesting congenital malformations, lymphatic obstruction, or acquired lymphatic proliferation as potential contributing factors. The clinical presentation of ovarian lymphangiomas often includes nonspecific symptoms such as abdominal pain, swelling, or discomfort, leading to difficulties in early detection and diagnosis. Radiological imaging, particularly Ultrasound, CT (computed tomography) and MRI (magnetic resonance imaging), plays a crucial role in identifying these lesions and guiding subsequent management strategies. Despite their generally benign nature, ovarian lymphangiomas can attain significant sizes, causing complications such as torsion, rupture, or compression of adjacent structures. Surgical intervention, typically in cystectomy or oophorectomy, is frequently pursued to alleviate symptoms and prevent potential complications. This paper aims to comprehensively review the existing literature on ovarian lymphangiomas, addressing their clinical presentation, diagnostic challenges, and management strategies. By synthesizing available data, we seek to enhance our understanding of this rare entity, providing valuable insights for clinicians encountering similar cases. Improved awareness and knowledge of ovarian lymphangiomas are essential for timely diagnosis and optimal patient outcomes.

Abstract Background Stathmin, a cytosolic microtubule-destabilizing phosphoprotein involved in the regulation of mitosis, is widely expressed in various malignancies and acts as an adverse prognostic factor. Our research analyzed its immunohistochemical expression on a large cohort of ovarian sex cord-stromal tumors, evaluating its potential utility in differential diagnosis, prognosis, and therapeutic application. Methods We examined 390 cases of ovarian sex cord-stromal tumors including 281 adult granulosa cell tumors (AGCT), 5 juvenile granulosa cell tumors (JGCT), 33 Sertoli-Leydig cell tumors (SLCT), 50 fibromas/thecomas (F/T), 11 Leydig cell tumors/steroid cell tumors (LCT/SterCT), 5 sex-cord stromal tumors NOS (SCST-NOS), 3 Sertoli cell tumors (SCT), and 2 sclerosing stromal tumors (ScST). Immunohistochemical analysis was performed using TMAs. Results Strong expression (> 50%) was observed in all cases of AGCT, JGCT, SLCT, SCST-NOS, SCT and 1 ScST. The other case of ScST exhibited mild expression (5–10%). The negative cases included exclusively F/T and LCT/SterCT, with F/T showing 24% of negative cases and LCT/SterCT comprising 64% of negative cases. Conclusion The results of our study indicate that stathmin is neither a prognostic marker nor suitable for the differential diagnosis of challenging cases of ovarian sex cord-stromal tumors. However, its predictive value may be theoretically significant, as a decrease in stathmin expression potentialy influences response to chemotherapy treatment.

Abstract The idea of utilizing unused oocytes present in the ovaries has been tested in various ways to produce offspring. However, only a limited number of studies succeeded in offspring generation. They include transplantation of ovaries into autologous or allogeneic animals, and acquisition of pups from oocytes obtained by transplanting mouse ovaries into immunodeficient rats. Here we report successful production of rat oocytes by transplanting rat ovaries under the kidney capsule of immunodeficient mice with addition of hormone administration to the mice. In addition, these oocytes were developed by in vitro fertilization, and transplanted into the oviducts of pseudopregnant rats, resulting in successful delivery of pups. The modified gene of the donor rat was confirmed to be correctly inherited to the pups. These results show that xenotransplantation of ovarian tissue makes it possible to leave offspring, beginning a new phase in developmental engineering.

Background: Infertility is a health problem that affects approximately 10-15% of couples trying to achieve pregnancy worldwide. Various studies have shown that Nigella sativa (N. Sativa) has many health benefits, including reproductive health. However, studies examining the effectiveness of N. Sativa in preventing folliculogenesis disorders have never been reported. Purpose: This study aims to determine the benefits of administering N. Sativa in preventing folliculogenesis disorders as measured by the diameter of the antral follicles and the thickness of the granulosa cell layer. Methods: This research design uses an experiment with a randomized posttest-only controlled group design approach. There were 27 experimental animals (mice) divided into three groups, namely the negative control group (9 animals) and the positive control group (9 animals), as well as the treatment group (9 animals). The positive control group was exposed to two cigarettes of cigarette smoke per day. The treatment group was exposed to cigarette smoke and N. Sativa 300 mg/KgBW/day for 28 days. After that, the mice were sacrificed to take their ovaries. Then, the ovarian organ samples were stained with Hematoxylin-eosin and examined for the diameter of the antral follicles and the thickness of the granulosa cell layer in the antral follicles with photo slides at 400x magnification in 5 fields of view, statistical analysis using the Independent T-test. Results: There were significant differences in antral follicle diameter and granulosa cell thickness in mice exposed to cigarette smoke and N. Sativa. Administration of N. Sativa extract can increase granulosa cell thickness through estrogenic function. Administration of N. Sativa extract showed estrogenic activity by increasing serum estradiol levels. N. Sativa heterotrophic activity cannot be separated from the flavonoid and phenolic compounds contained in N. Sativa, which have been proven to have high estrogenic activity. Conclusion: N. Sativa extract is efficacious in improving the diameter of antral follicles and the thickness of granulosa cells caused by exposure to cigarettes.

Abstract A 27-year-old man was admitted to the hospital after a year of marriage due to infertility. During laparoscopic exploratory surgery, the presence of a retrovesical uterus was clearly observed, and the gonadal organs were visible on both sides. However, the testicles or ovaries were not identifiable, nor were the spermatic vessels and fallopian tubes at the joint. Intraoperative bilateral gonad biopsy was performed. Cryopreservation of the right gonadal gland revealed gonadoblastoma and malignant germinoma (asexual tumor/seminoma) with sclerosis and atrophy of testicular tissue. No proliferation of germ cells and sertoli cells was observed in spermatic tubule. The left gonad was diagnosed as a gonadoblastoma. Finally, total hysterectomy and bilateral gonadal tumor organectomy were performed to seal the vaginal stump. Local radiotherapy was administered after surgery. In general, tumors were found on both sides of the gonads, especially gonadoblastoma and malignant germinoma on the right side and gonadoblastoma on the left side.

Abstract Background Secreted frizzled-related proteins (SFRPs) comprise a family of WNT signaling antagonists whose roles in the ovary are poorly understood. Sfrp4-null mice were previously found to be hyperfertile due to an enhanced granulosa cell response to gonadotropins, leading to decreased antral follicle atresia and enhanced ovulation rates. The present study aimed to elucidate the mechanisms whereby SFRP4 antagonizes FSH action. Methods Primary cultures of granulosa cells from wild-type mice were treated with FSH and/or SFRP4, and effects of treatment on gene expression were evaluated by RT-qPCR and RNAseq. Bioinformatic analyses were conducted to analyse the effects of SFRP4 on the transcriptome, and compare them to those of FSH or a constitutively active mutant of FOXO1. Additional granulosa cell cultures from wild-type or Sfrp4-null mice, some pretreated with pharmacologic inhibitors of specific signaling effectors, were used to examine the effects of FSH and/or SFRP4 on signaling pathways, autophagy and apoptosis by western blotting and TUNEL. Results Treatment of cultured granulosa cells with recombinant SFRP4 was found to decrease basal and FSH-stimulated mRNA levels of FSH target genes. Unexpectedly, this effect was found to occur neither via a canonical (CTNNB1-dependent) nor non-canonical WNT signaling mechanism, but was found to be GSK3β-dependent. Rather, SFRP4 was found to antognize AKT activity via a mechanism involving AMPK. This lead to the hypophosphorylation of FOXO1 and a decrease in the expression of a portion of the FSH and FOXO1 transcriptomes. Conversely, FSH-stimulated AMPK, AKT and FOXO1 phosphorylation levels were found to be increased in the granulosa cells of Sfrp4-null mice relative to wild-type controls. SFRP4 treatement of granulosa cells also induced autophagy by signaling via AKT-mTORC1-ULK1, as well as apoptosis. Conclusions This study identifies a novel GSK3β-AMPK-AKT signaling mechanism through which SFPR4 antagonizes FSH action, and further identifies SFRP4 as a novel regulator of granulosa cell autophagy. These findings provide a mechanistic basis for the phenotypic changes previously observed in Sfrp4-null mice, and broaden our understanding of the physiological roles of WNT signaling processes in the ovary.

Abstract Objectives We present an Egyptian study on pediatric ovarian immature teratomas (ITs), aiming to clarify our treatment strategy selection. Methods A retrospective review of all children with pure ovarian ITs who were treated at our institution between 2008 and 2023. The analysis included clinical characteristics, tumor staging according to Children’s Oncology Group (COG), grading based on the Norris system, management, and outcomes. Results Thirty-two patients were included, with a median age of 9 years. All patients underwent primary surgery. Unilateral salpingo-oophorectomy was performed in 31 patients. Surgical staging was completed in all patients. Based on COG staging, there were 28 patients (87.5%) stage I, 1 (3%) stage II, and 3 (9.5%) stage III. According to Norris classification, 16 patients (50%) were classified as grade I, 9 (28%) grade II, and 7 (22%) grade III. All patients in stage I were treated using surgery-alone approach, whereas the remaining four (12.5%) received adjuvant chemotherapy. Five patients in stage I had gliomatosis peritonei (GP), and none of them underwent extensive surgery. At a median follow-up of 86 months, two patients had events. The first patient (stage III/grade I) developed IT relapse on the operative bed, and the second (stage I/grade I) had a metachronous IT on the contralateral ovary. Both patients were successfully managed with surgery followed by second-line chemotherapy. Five-year overall survival and event-free survival for all patients were 100% and 93.4%, respectively. Conclusions Surgery-alone strategy with close follow-up achieves excellent outcomes for localized ovarian ITs in children, irrespective of the Norris grading or the presence of GP. However, adjuvant chemotherapy is questionable for patients with incompletely resected or locally advanced tumors, and its role requires further evaluation through prospective multicentric studies with a larger sample size.

Ovarian carcinoid is a rare well-differentiated neuroendocrine tumor resembling those arising in the gastrointestinal tract. We present a case of ovarian carcinoid with magnetic resonance imaging (MRI) findings. A 50-year-old woman with genital bleeding and severe constipation was referred to our hospital. On MR imaging, a left ovarian tumor showed iso to high signal intensity on T1-weighted images (T1WI), relatively low signal intensity on T2WI, and slightly high signal intensity on diffusion-weighted images. Additionally, the tumor demonstrated early and delayed strong contrast enhancement on dynamic contrast-enhanced images. The tumor was pathologically diagnosed with ovarian strumal carcinoid. High signal intensity on T1WI should be recognized as the MRI findings in ovarian carcinoids.

Juvenile granulosa cell tumour (JGCT) is very uncommon gynecological malignancy that occurs more commonly in under five years old of age. We describe a case of JGCT in a 4-years old girl. The malignancy is assigned to International Federation of Gynecology and Obstetric staging system (FIGO stage I). Treated with complete excision only, the patient showed no evidence of relapse one year after surgery. Findings in this case are discussed and histological examination confirmed the diagnosis. The naturalhistory of JGCT, epidemiology, histology, treatment and prognosis are reviewed along with the case presentation.

The present study aimed to identify the novel mechanisms regulating rabbit fecundity. For this purpose, the association between litter size variability, fecundity, ovarian morphology, and markers of proliferation, apoptosis, steroidogenesis, and the presence of regulatory proteins were examined in ovarian cells from two rabbit lines divergently selected for low (LL) and high (HL) variability in litter size throughout sixteen generations. Ovaries and uteri were isolated from multiparous non-lactating female rabbits from each line at sixteen generations of selection. One part of the ovary was subjected to histomorphometric analysis of folliculogenesis. From the rest of ovary, ovarian granulosa cells (OGCs) were isolated, cultured, and cell viability, proliferation (accumulation of PCNA and of cyclin B1, and BrdU-positive cells), and apoptosis (accumulation of caspase 3, bax and DNA fragmentation) were evaluated by the Trypan blue exclusion test and BrdU, quantitative immunocytochemistry, and cell death detection assays. Furthermore, OGCs were subjected to proteomic analysis by using the nano HPLC-Chip-MS/MS method. The release of progesterone and oestradiol was measured by ELISA. The LL had more than one kit per litter than the HL (7.6 kits vs. 6.5 kits, p ≤ 0.05). No differences were found in the diameter of primordial and primary ovarian follicles, theca, and granulosa thickness, but the diameter of oocytes in the primary and secondary follicles was higher in the LL than in the HL (88.49 µm in the LL vs. 77.86 µm in the HL for oocytes of primary follicles, p ≤ 0.05; 122.10 µm in the LL vs. 109.87 µm in the HL for oocytes of secondary follicles, p ≤ 0.05). Preovulatory follicles were presented only in the ovaries of the LL. The LL had higher incorporation of BrdU and reduced accumulation of bax within OGCs (0.92% in the LL vs. 0.44% in the HL for incorporation of BrdU, p ≤ 0.05; 41% in the LL vs. 48% in the HL for accumulation of bax, p ≤ 0.05). Ovarian fragments from the LL produced less progesterone and oestradiol than those of the HL (12 ng/mg tissue/day in the LL vs, 45 ng/mg tissue/day in the HL for progesterone, p ≤ 0.05; 11 ng/mg tissue/day in the LL vs. 30 ng/mg tissue/day in the HL for oestradiol, p ≤ 0.05). Besides, the OGCs from the LL produced a higher number of specific regulatory proteins involved in cell differentiation, proliferation, and adhesion than the HL (50 vs 38, p ≤ 0.05). In conclusion, higher prolificacy in the LL line would be caused by: (1) the selection of growing primordial ovarian follicles; (2) better transformation to preovulatory follicles; (3) increased cytoplasmic maturation of oocytes; (4) increased DNA synthesis and decreased cytoplasmic apoptosis in OGCs; (5) changes in ovarian steroidogenesis; and (6) changes in the number of peptides involved in cell differentiation, proliferation, and adhesion.HIGHLIGHTS Assessing fecundity in two divergently rabbit lines for variability in litter size. Fecundity and its variability are related to the diameter and quality of oocytes. Selection and better growth of preovulatory follicles have been related to fecundity.

AbstractChinese herbs contain substances that regulate female hormones. Our study confirmed that Zingiberis rhizoma carbonisata contains Zingiberis rhizoma-based carbon dots (ZR-CDs), which exert regulatory effects on serum oestradiol and FSH in mice and show impacts on endometrial growth and follicular development that potentially affect the ability of female fertility. ZR-CDs were characterized to clarify the microstructure, optical features, and functional group characteristics. It shows that ZR-CDs are spherical carbon nanostructures ranging from 0.97 to 2.3 nm in diameter, with fluorescent properties and a surface rich in functional groups. We further investigated the impact of ZR-CDs on oestradiol and FSH in serum, growth, and the development of ovarian and uterine using normal female mice and exogenous oestradiol intervention model. It was observed that ZR-CDs accelerated oestrogen metabolism and attenuated oestradiol-induced endometrial hyperplasia. Simultaneously, ZR-CDs triggered an increase in FSH, even in the presence of high-serum oestradiol that inhibits FSH secretion. Our findings suggest that ZR-CDs could be a potential therapeutic treatment for anovulatory menstruation.

The Pacific white shrimp Penaeus vannamei is one of the most productive shrimp species in China and the world. Domestic demand for P. vannamei has increased annually because of its fast growth, easy breeding, strong resistance to stress and disease, and high meat yield. The primary challenge in the development of the shrimp industry is seed rearing. The demand for broodstocks of P. vannamei in China is as high as 1, 000, 000 pairs per year, and the total weight of shrimp seedlings exceeds 1.3 trillion. China needs to import 300, 000 pairs of P. vannamei every year. The cost of each pair of P. vannamei is approximately 200 US＄, thus equating to hundreds of millions of US＄ in total. During the production of P. vannamei, the spawning frequency varies greatly among female shrimp. Cultivating high-fecundity P. vannamei through genetic improvement is an effective way to reduce the cost of seedling farms and improve economic benefits.To further study the molecular mechanisms of fecundity differences in female P. vannamei, the ovarian histology and expression of four candidate genes in populations with high- and low-fecundity were analyzed in this study. Before the experiment, the 7-month-old parent shrimp of P. vannamei were fed a commercial parent shrimp fortified diet, a maturation promoting diet, and squid for nutritional enhancement. After one month of promoting maturity, the unilateral eyestalk of female shrimp was cut using forceps, and next-generation family construction began after 15–20 days of recovery. The spawning frequency of 439 female shrimp in the 30-day production cycle was calculated and used to indicate fecundity. Those with a spawning frequency of 0 were included in the low-fecundity group, while those with a spawning frequency of > 3 were included in the high-fecundity group. The ovarian development of P. vannamei was divided into four stages (proliferation, small growth, large growth, and maturation) according to the color, size, and shape of the ovaries. In the high-fecundity group, the ovaries exhibited development up to stageⅣ, indicating their attainment of maturity. In the low-fecundity group, ovarian development was limited to stageⅢ, with progression to stageⅣ proving difficult. Ovarian tissues from the same part of female shrimp of stageⅠ–Ⅳ in the high-fecundity group and stageⅠ–Ⅲ in the low-fecundity group were taken and immediately immersed in tissue fixative for paraffin sectioning. The research team previously identified candidate genes related to fecundity in genomic regions undergoing selective sweep in isolated populations. These included thioredoxin 2 (TRX2), partitioning-defective 3 (PARD3), phospholipase Cβ4 (PLCβ4), and arginine-glutamate acid dipeptide repeats (RERE), which play important roles in oocyte maturation, cell proliferation, and early development in other species. qRT-PCR was used to analyze and compare their expression in the ovary and eyestalk tissues of high- and low-fecundity females at different stages of ovarian development.The results of paraffin section showed that the ovarian development of the low-fecundity group was slower than that of the high-fecundity group. In the high-fecundity group, stageⅠ ovaries primarily consisted of oogonia and oocytes during the early phase of yolk formation, with the emergence of small spherical yolk granules. StageⅠ ovaries in the low-fecundity group primarily consisted of oogonia, with no observable yolk granule formation. StageⅡ ovaries in the high-fecundity group were mainly composed of oocytes with vitelline formation, accompanied by a notable augmentation in both the size and number of yolk granules. StageⅡ ovaries in the low-fecundity group primarily comprised oocytes in the early phase of vitelline formation with yolk granules beginning to form. StageⅢ ovaries in the high-fecundity group consisted mainly of oocytes during the late phase of yolk formation. Many spherical yolk granules were formed in the cytoplasm, and cortical rods began to appear, although the lengths of the cortical rods were short. StageⅢ ovaries in the low-fecundity group were composed of oocytes during yolk formation. The cytoplasm was full of spherical yolk granules in the absence of cortical rods. In the high-fecundity group, stage Ⅳ ovaries were filled with mature oocytes, large yolk granules filled the entire cytoplasm, and thick cortical rods were formed in the oocytes. qRT-PCR showed that the four genes maintained high expression levels in the mature ovaries of the high-fecundity group. Their expression levels in the ovaries of the high-fecundity group were higher than those in the ovaries of the low-fecundity group in the small and large growth stages. TRX2, PLCβ4, and RERE expression levels were significantly different between the groups (P < 0.05). In eyestalk tissues at different stages, TRX2, PARD3, PLCβ4, and RERE expression levels in the low-fecundity group were higher than those in the high-fecundity group. The results suggest that these four genes may play important roles in the ovarian development of P. vannamei. The expression levels of all four genes in the high-fecundity group were lower than those in the low-fecundity group in eyestalk tissues. However, in ovarian tissues, except for the PARD3 gene in stageⅠ, which was expressed at lower levels in the high-fecundity group than in the low-fecundity group, the expression levels of all four genes in the other stages were higher in the high-fecundity group than those in the low-fecundity group, which is opposite to the expression trend in eyestalk tissues. Therefore, we speculate that these genes also promote cell proliferation, development, and target gene expression in the eyestalks of the low-fecundity group. Their high expression in the eyestalks of the low-fecundity group may enhance the secretion of ovarian inhibitory factors, thereby inhibiting ovarian development. Further research is needed to reveal their functional mechanisms. The above results provide an important reference for the in-depth study of the molecular mechanisms of differences in the fecundity of female P. vannamei.

Background and purpose: Paraquat is one of the chemical herbicides with high toxicity. In case of incorrect consumption, this chemical causes fatal poisoning in people, especially those active in the field of agriculture. Nowadays, using compounds of natural origin to improve or reduce the harmful effects of agricultural pesticides is one of the important issues that has attracted the attention of researchers in this field. Quercetin as a natural flavonoid is found in large quantities in fruits and vegetables. As a kind of natural antioxidant, this compound has great power in removing free radicals. The purpose of this research was to evaluate the protective effects of quercetin on ovarian toxicity caused by paraquat in rats. Materials and methods: To conduct this experimental research, 30 adult Wistar rats with an average age of 6 to 8 months and a weight range of 160 to 180 gr were randomly divided into 5 experimental groups including 6 rats in each group. Paraquat (1.25 mg/kg, intraperitoneally) was injected into mice. Then quercetin was injected intraperitoneally in concentrations of 50, 100, and 200 mg/kg. The negative control group received only normal saline. After 24 hours, the animals were sacrificed and their ovaries were evaluated for changes in oxidative stress indices. Results: The results showed that paraquat significantly increased the reactive oxygen species (ROS), protein carbonyl, and lipid peroxidation, and significantly decreased glutathione content and survival of ovarian tissue cells when compared with the control group (P

Clock genes play a pivotal role in the rhythm maturation of bony fish ovaries. To explore the role of clock genes in the rhythmic ovarian development and maturation of tongue sole (Cynoglossus semilaevis), we used real-time fluorescent quantitative PCR technique to analyze the expression profiles of circadian locomotor output cycles Kaput 1a (Clock1a), brain and muscle Arntlike 1a (Bmal1a), Cryptochrome 1a (Cry1a), Cryptochrome 2 (Cry2), and Period 2 (Per2) at the ovarian stages Ⅱ, Ⅲ, Ⅳ, Ⅴ, and Ⅵ.The coding DNA sequence (CDS) of five clock genes were cloned and phylogenetically analyzed. We found that the CDS sequence length of Clock1a was 1 620 bp and encoded 539 amino acids, with the encoded amino acid sequence of Clock1a having a predicted molecular weight of 81.9 kDa. Clock1a has the functional domain PASD1 (consisting of 64 amino acids) and PAS11 (consisting of 103 amino acids). The CDS sequence length of Bmal1a was 1 881 bp and encoded 626 amino acids, with the encoded amino acid sequence of Bmal1a having a predicted molecular weight of 68.9 kDa. In the Bmal1a sequence, the functional domain PASD3 is composed of 63 amino acids, and the functional domain PAS11 is composed of 103 amino acids. The CDS sequence length of Cry1a was 1 896 bp and encoded 631 amino acids, with the encoded amino acid sequence of Cry1a having a predicted molecular weight of 71.4 kDa. Its functional domain FAD7 was composed of 199 amino acids. The CDS sequence length of Cry2 was 2 007 bp and encoded 669 amino acids, with the encoded amino acid sequence of Cry2 having a predicted molecular weight of 76.0 kDa. The Cry2 sequence contains a functional domain FAD7 consisting of 199 amino acids. The CDS sequence length of Per2 was 4 248 bp, encoding 1 415 amino acids, and the encoded amino acid sequence of Per2 has a predicted molecular weight of 154.0 kDa. In the Per2 sequence, the functional domain PeriodC consists of 295 amino acids and a functional domain PAS11 consists of 102 amino acids.The neighbor-joining method was used to analyze the Clock1a, Bmal1a, Cry1a, Cry2, and Per2 phylogenetic relationships between C. semilaevis and other bony fish, amphibians, birds, and mammals. The homology of Clock1a, Bmal1a, Cry1a, Cry2, and Per2 with other bony fish was 60%–79%, 94%–100%, 85%–91%, 84%–94%, and 70%–84%, respectively. Therefore, we believe that these five amino acid sequences show strong conserved property. In addition, the homology of Cry1a and Cry2 was 62%, indicating that Cry1a and Cry2 evolved differently during the evolution of C. semilaevis. In the constructed phylogenetic tree, Clock1a, Bmal1a, Cry1a, Cry2, and Per2 of C. semilaevis were clustered together with other bony fishes, indicating a close relationship between C. semilaevis and other bony fish in the evolutionary process. Moreover, the homology of Clock1a between C. semilaevis and mammals, birds, and amphibians is low, indicating that there are evolutionary differences in the evolutionary process of Clock1a. The high homology of Bmal1a, Cry1a, Cry2, and Per2 with mammals, birds, and amphibians suggests that these four clock genes were strongly conserved during the evolution of C. semilaevis.In the present study, we found that the expression levels of five clock genes were high in stagesⅡ and Ⅲ, which were equivalent to the non-reproductive season (P < 0.05). However, the expression levels of five clock genes were low in stages Ⅳ and Ⅴ, which were equivalent to the reproductive season (P < 0.05). Therefore, the expression profiles of Clock1a, Bmal1a, Cry1a, Cry2, and Per2 in the ovaries of C. semilaevis also have seasonal characteristics. The ovarian development and maturation of C. semilaevis goes through stages Ⅱ, Ⅲ, Ⅳ, Ⅴ, and Ⅵ, and then reaches stage Ⅱ again and starts a new reproductive cycle. The variation patterns of seasonal factors, such as light and temperature, in fish ovaries were consistent with the annual expression patterns of the five clock genes in this study. Therefore, it can be considered that the expression of clock genes in C. semilaevis has an annual cycle. The findings presented in this study can enrich the theory of ovarian development and maturation of C. semilaevis and provide a theoretical basis for improving breeding technology and seedling efficiency.

Abstract Background The common marmoset, Callithrix jacchus, is an invaluable model in biomedical research. Its use includes genetic engineering applications, which require manipulations of oocytes and production of embryos in vitro. To maximize the recovery of oocytes suitable for embryo production and to fulfil the requirements of the 3R principles to the highest degree possible, optimization of ovarian stimulation protocols is crucial. Here, we compared the efficacy of two hormonal ovarian stimulation approaches: 1) stimulation of follicular growth with hFSH followed by triggering of oocyte maturation with hCG (FSH + hCG) and 2) stimulation with hFSH only (FSH-priming). Methods In total, 14 female marmosets were used as oocyte donors in this study. Each animal underwent up to four surgical interventions, with the first three performed as ovum pick-up (OPU) procedures and the last one being an ovariohysterectomy (OvH). In total, 20 experiments were carried out with FSH + hCG stimulation and 18 with FSH-priming. Efficacy of each stimulation protocol was assessed through in vitro maturation (IVM), in vitro fertilization (IVF) and embryo production rates. Results Each study group consisted of two subgroups: the in vivo matured oocytes and the oocytes that underwent IVM. Surprisingly, in the absence of hCG triggering some of the oocytes recovered were at the MII stage, moreover, their number was not significantly lower compared to FSH + hCG stimulation (2.8 vs. 3.9, respectively (ns)). While the IVM and IVF rates did not differ between the two stimulation groups, the IVF rates of in vivo matured oocytes were significantly lower compared to in vitro matured ones in both FSH-priming and FSH + hCG groups. In total, 1.7 eight-cell embryos/experiment (OPU) and 2.1 eight-cell embryos/experiment (OvH) were obtained after FSH + hCG stimulation vs. 1.8 eight-cell embryos/experiment (OPU) and 5.0 eight-cell embryos/experiment (OvH) following FSH-priming. These numbers include embryos obtained from both in vivo and in vitro matured oocytes. Conclusion A significantly lower developmental competence of the in vivo matured oocytes renders triggering of the in vivo maturation with hCG as a part of the currently used FSH-stimulation protocol unnecessary. In actual numbers, between 1 and 7 blastocysts were obtained following each FSH-priming. In the absence of further studies, FSH-priming appears superior to FSH + hCG stimulation in the common marmoset under current experimental settings.

Malathion, a widely used organophosphate, is known for its relatively low toxicity and extensive application. However, it has been found to act as a female reproductive toxicant by causing oxidative stress, apoptosis, autophagy, and hormonal imbalances. Hesperidin, a flavonoid belonging to the flavanone class, exhibits various beneficial properties such as antioxidant and anti-inflammatory effects, which can potentially counteract harmful effects. The objective of this study was to examine how hesperidin and malathion impact the expression of miRNAs and genes linked to apoptosis and inflammation. Balb/c mice (n = 40) were divided into four groups: hesperidin (20 mg/kg), malathion (3 mg/kg), hesperidin+malathion, and control. After a 35-day intraperitoneal treatment, the mice were sacrificed. The left ovaries were used for analyzing the expression of miRNA-146a-5p, miRNA-129-3p, miRNA-96-5p, NF-κB, Bax, and Bcl-2 through RT-qPCR, as well as the levels of several cytokines using the ELISA method. The right ovaries were examined through histological and immunohistochemical techniques using H&E and NF-κB staining. Malathion exposure led to an increased Bax/Bcl-2 ratio, upregulated expression of Bax and NF-κB, elevated levels of IFN-γ, IL-2, and IL-6, enhanced expression of miRNA-146a-5p, decreased expression of miRNA-129-3p and miRNA-96-5p, and reduced levels of IL-4 and IL-10. Additionally, malathion-exposed ovaries exhibited structural abnormalities and disrupted architecture, accompanied by heightened NF-κB immunoreactivity. Conversely, treatment with hesperidin showed its capacity to counteract the detrimental consequences of malathion on the ovaries by alleviating or reversing these changes. In conclusion, hesperidin showed protective effects against malathion-induced ovarian toxicity by modulating cytokine production, apoptosis, inflammation, and miRNA expression.

Aim – to study the dynamics of ovarian parameters in women from the first period of adulthood to early old age measured by sonography. Material and methods. The study used the results of an ultrasound examination of 81 women with normal pelvic dimensions, without reproductive organ pathology, having maximum two pregnancies in life history more than one year before the study. All participants gave their consent to routine examination. The procedure consisted of measuring the length, width, anteroposterior size and volume of the ovaries by transvaginal access according to standard protocols. The subjects were divided into three groups according to anatomical age classification. The first group consisted of 29 early adulthood women (22 to 35 years); the second group included 23 women in middle adulthood (36 to 56 years) and 29 early old age women (57 to 73 years). Results. All the ovarian parameters decreased by early old age (p 0.01). At the same time, a trend towards macroanatomic asymmetry of the ovaries with larger dimensions in the right side was revealed in each of the studied age periods (p 0.05). Conclusion. The results of a lifetime comparative analysis of the anatomical dimensions of the ovaries in women contribute to the knowledge of their age-specific structure and requires a future detailed study. A new knowledge is required to develop modern methods for preventing pathologies of the female reproductive system and to provide timely, high-quality medical care.

Ovarian transposition (OT) has been proposed as a protective measure against radiation-induced damage to ovarian function and fertility. Despite its historical use, limited research has focused on evaluating endocrine and exocrine ovarian function after OT performed in adolescents and young adults (AYAs) before or during puberty. The purpose of our study was to investigate the fertility, pubertal development, and ovarian function of women with a previous history of OT during childhood, adolescence or young adulthood. In an observational bicentric retrospective study, we included 32 young female cancer patients who underwent OT before the age of 26 between 1990 and 2015 at Lyon Léon Bérard Cancer Center or Nancy University Hospital. The mean age at the time of OT was 15.6 years with a cancer diagnosis at 15 ± 4.8 years. Among the 10 women attempting pregnancy post-treatment, 60% achieved successful pregnancies. After a mean follow-up of 9.6 ± 7 years, 74% (17 out of 23) of women recovered spontaneous menstrual cycles (seven out of eight evaluable women with OT before or during puberty). Notably, 35% of women who did not attempt pregnancy demonstrated adequate ovarian reserve. Ovarian reserve and function recovery were influenced by the specific chemotherapy received. Importantly, our findings suggest that OT’s effectiveness on ovarian activity resumption does not significantly differ when performed before or during puberty compared to pubertal stages. This study contributes valuable insights into the long-term reproductive outcomes of young women undergoing OT, emphasizing its potential efficacy in preserving ovarian function and fertility across different developmental stages.

Objective To investigate the effect of membrane-spanning 4-domains subfamily member 6D (MS4A6D) gene knockout on the fertility of female mice. Methods On the basis of the successful construction of Ms4a6d gene knockout model mice, the homozygous Ms4a6d gene knockout (Ms4a6d-/-) mice at an age of different weeks were subjected. Mouse genotype was identified with RT-qPCR and agarose gel electrophoresis. HE staining, immunofluorescence assay and ELISA were used to detect serum level of anti-müllerian hormone (AMH), composition of macrophages and follicles at all levels in the ovaries of Ms4a6d-/- mice, respectively. The fertility changes in pregnant rate and average litter size of female Ms4a6d-/- mice were observed by fertility experiments. Results Compared with the wild-type (Ms4a6d+/+) female mice at the same age, there were less macrophages in the ovarian tissues of the 2- and 4-week-old Ms4a6d-/- female mice (P < 0.01), but no significant difference was observed between the 2 types of mice at 8 weeks of age. For the adult Ms4a6d-/- female mice at an age of 8 weeks, the ovarian coefficient was obviously lower (P < 0.01), and the numbers of primordial, primary, secondary and antral follicles in the ovarian tissues was statistically lower (P < 0.05) when compared with the adult wild-type female mice at the same age. The serum AMH level was notably lower in the Ms4a6d-/- female mice of all ages than the wild-type ones at corresponding ages (P < 0.05). The average pregnancy rate was 56% and the litter size was 4.1±1.1 in adult Ms4a6d-/- female mice, which were significantly lower than those of wild-type mice of the same age (89% and 6.3±1.2). Conclusion Ms4a6d gene knockout decreases the number of macrophages in the ovarian tissues of adolescent females, interferes with the development of follicles in the ovarian tissues of adolescent females, and leads to a decrease in ovarian reserve in adult females and a decrease in fertility in adult females.

Objective: The effects of Oviductus Ranae (OR) on cyclophosphamide-induced follicular developmental disorders were analyzed from the perspective of transcriptomics, which provided sufficient theoretical and experimental basis for the study of estrogen-like mechanism of OR. Methods: Sixty screened female Wistar rats were randomly divided into control group (C), model group (M), positive group (P), OR low-dose (ORL) and high-dose group (ORH). Except for the control group, all rats were injected with cyclophosphamide intraperitoneally to establish the rat model of follicular development dysfunction. Except for the model group, the other rats were given corresponding drugs by gavage, and the serum sex hormone contents were determined after 14~16 days of continuous gavage. The ovaries and uterus were extracted and weighed for recording and calculating the organ index. One ovary and uterus were stained with HE staining to calculate the number of follicles at all levels and the thickness of the endometrium, while the other ovary was analyzed by transcriptome, and the differentially expressed genes were screened and analyzed by GO and KEGG enrichment analyses. Results: The results showed that the rat follicular development dysfunction model was successfully established. Compared with the rats in the M, the wet weight and index of the uterus of rats in the ORL and ORH significantly increased (P

Urotensin-I (UI) is a peptide belonging to the corticotropin-releasing hormone family, produced in the urophysis of teleosts. This investigation was undertaken to examine the effect of UI on the hypothalamo-hypophyseal-ovarian axis in the Mozambique tilapia, a cichlid fish. In the first experiment, administration of UI (2 μg/100 μL saline) for 22 days caused a significant reduction in the number of follicles at stages I, II, III (previtellogenic), IV (vitellogenic), and V (preovulatory), concomitant with a significant increase in the percent atresia of follicles at stages II, III, and IV than in controls. In the pituitary, gonadotropin-releasing hormone (GnRH) - positive fibres were detected in the proximal pars distalis (PPD) in both controls and UI-treated fish; however, there was a significant reduction in the intensity and percent area of GnRH-immunoreactive fibres in fish treated with UI compared with controls. The serum luteinizing hormone (LH) level was significantly lower, whereas a significant rise in the serum level of cortisol was noticed in UI-treated fish compared with controls. In the second experiment, treatment with a low dose of UI (10 mM) did not affect the number of ACTH-secreting cells in vitro, but the number of these cells was significantly higher in high-dose UI (50 mM)-treated pituitaries compared to controls. Together, the present investigation reveals for the first time that the treatment of UI affects the follicular development process through the downregulation of the GnRH-LH axis and activation of the pituitary-interrenal pathway in teleosts.

Among vertebrate species, fishes are considered to be the most developmentally diverse taxa exhibiting remarkable gonadal plasticity with capacity of sex reversal. Sex determination in teleost fishes is governed via a complex regulatory network involving multiple genetic, environmental (temperature, population density, pH), social interaction/stress factors. Piscine germ-line originates as Primordial germ cells (PGC) from cleavage blastomeres/endoderm and gets specified by maternal inheritance of Vasa, Nanos, Tdrds, Dnd, Dazl etc and subsequently migrates to reach the embryonic gonadal primordium. Post-hatching, mitotically active germ cells (Gc) continue to proliferate via two distinct pathways-firstly, stem cell-like mode followed by meiosis-committed cystic division. In Japanese rice fish medaka, Oryzias latipes sex gets determined on the chromosomal basis, where XY becomes a male because of testicular Dmy transcription and XX develops as female owing to Gc restricted Foxl3 expression. Sex specific germ-line stem cell systems also have been found in adult O. latipes. Intriguingly in teleost fishes, the developmental fate of transplanted Gc do not depend on the sex of the donor rather solely relies upon the sex of the recipient. Furthermore, sexually mature piscine gonads remain developmentally plastic showing extensive responsiveness towards sex steroids. The critical regulation of gonadal differentiation is directed by female specific 17β-estradiol (E2) and bio-active male androgen 11-keto-testosretone (11-kT). Various transcription factors (TFs) e.g.- Dmrt1, DAX1, Ad4BP/SF-1, Foxl2 regulate the bio-conversion rate of testosterone (T) to either E2 or 11-kT. Similarly, multiple endocrine factors (EFs) e.g.- LH, FSH, E2, 11-kT and growth factors (GFs) e.g- AMH, GSDF, IGF-1/3, GDF-9/BMP-15 critically induce Gc differentiation to regulate the gametogenic output. In this article we have precisely discussed the endocrine regulation of Gc development during gonadal maturation of teleosts.

INTRODUCTION: Syringic acid (SA), a natural phenolic substance, possesses anti-inflammatory and antioxidant properties. We investigated the likely preventive effects of SA on organ damage to the reproductive system caused by bilateral ovarian ischemia-reperfusion (I/R). METHODS: 32 female Sprague-Dawley rats in total were used; they were split into four groups: a sham group for control (group I), I/R group (group II), I/R group given 10 mg/kg SA (group III), and an I/R group given 50 mg/kg SA (group IV). Group I underwent only an abdominal incision without I/R. In groups II, III, and IV, I/R procedures were conducted for 3 hours. SA was given intraperitoneally to groups III and IV for 30 minutes before reperfusion at 10 mg/kg and 50 mg/kg, respectively. The ovarian tissues were surgically removed for analysis. RESULTS: TNF-α, MDA, IL-1β, MPO, TOS and OSI levels in group II were found to be significantly higher than in group I. Additionally, a decrease was detected in TAS and SOD values in group II. In contrast, SA administration led to significant increases in the level of antioxidant enzymes in both groups III and IV. Also, in the SA-given groups, significant decreases were detected in OSI and TOS levels, along with a decrease in MPO, MDA, TNF-α, and IL-1β levels. DISCUSSION AND CONCLUSION: It has been demonstrated that SA has a protective effect against organ damage caused by bilateral ovarian I/R in the reproductive system. This is the first study to show that SA can reduce ovarian damage due to I/R injury.

Objectives: Ovarian torsion is an important gynaecological emergency because it causes ischemia in ovary. Our aim is to research the effects of Hypericum perforatum on p53, TNF-α, CAT, SOD, GSH, PTEN and is haPI3K/Akt/mTOR pathway in ovarian of rats with ischemia reperfusion (I/R) injury. Materials and Methods: The study included 56 adult female rats which were allocated to 7 groups; control, ischemia, ischemia 300HP, ischemia 600HP, I/R, I/R 300HP and I/R 600HP. H. perforatum was applied to ischemia 300HP, ischemia 600HP, I/R 300HP and I/R 600HP groups at the dose of 300 mg/kg or 600 mg/kg by oral gavage. Collected ovarian tissues were examined by light microscopy, biochemical and real-time PCR techniques. Results: In histological examination, the least degeneration was seen in 600HP group among other groups, apart from control group. TUNEL results showed that apoptosis level of other groups was lower than that of ischemia and I/R groups. CAT and GSH levels of I/R 600HP group increased. The highest level of PTEN and mTOR was found in the ischemia group. The Akt level was lower in I/R and I/R 300HP groups but its level was close to that of control and I/R 600HP groups. TNF-α and p53 mRNA expressions in I/R group were increased. Conclusions: The result of the study showed that H. perforatum (600 mg/kg) had an effect on tissue degeneration, CAT and GSH levels, and also TNF-α and p53 mRNA expressions levels. We suggest that H. perforatum can be considered as an agent to protect against tissues damage during ischemia reperfusion.

Practically, when only parts of an unidentified human body are found, age estimation with limited materials is required. The purpose of this study was to investigate methods for estimating age using the uterus and ovaries. Among forensic autopsies performed between January 2011 and March 2022, 211 uteruses and 521 ovaries of 322 women were used for this study. Measured values for ovarian weight and uterine volume index were corrected by body surface area to consider the effect of body size. The corrected uterine volume index increased in teenage years and achieved its maximum in the 40–49 group, then gradually decreased with increasing age. The corrected ovarian weight increased until the twenties, after which it decreased with age. For women aged 20 years or more, receiver operating characteristic (ROC) curve analysis suggested that a uterine volume index of 41.2 cm3/m2 was the cutoff value for classifying the age as ≥60 years or 2 and 1.92 g/m2 were the cutoff values for classifying the age as ≥40 years or

The goal of this study was to investigate the effects of Ferula communis L. on growth performance, relative organ weights, tissue and serum oxidative status, and biochemical and histopathological changes in rats exposed to continuous light. Rats exposed to continuous light for 10 days were given F. communis orally. Daily body weight was recorded, and rats were euthanized by cervical dislocation. Blood was collected to determine relative organ weights, followed by immediate biochemical and histopathological analysis of the organs. Serum and tissue oxidative status were measured. Continuous light exposure in rats resulted in weight loss, decreased ovary, uterus, and kidney weights, diminished total antioxidant status (TAS) along with increased cortisol, total oxidant status, and oxidative stress index. However, F. communis treatment reduced serum cortisol concentration and alleviated oxidative stress by increasing TAS even under prolonged light exposure conditions. Neither continuous light exposure nor F. communis treatment caused significant changes in malondialdehyde and glutathione in organ tissues. Although continuous light caused an increase in the number of cystic follicles, F. communis treatment did not seem to have a positive effect on cystic follicle formation. In conclusion, continuous light exposure stressed the rats and increased cortisol, as well as stimulated oxidative stress and cystic follicle formation. F. communis treatment can help alleviate the harmful effects of constant light exposure.

This study aimed to optimize the composition of a liquid medium for Mycoplasma gallisepticum growth rate and density through statistical approach. The growth concentration of M. gallisepticum was determined by plotting a standard curve using Real-Time Quantitative PCR. The one-factor-at-a-time method and the Plakett-Burman design were initially applied to identify the factors that influenced the biomass of M. gallisepticum. The steepest ascent experiment and response surface methodology (RSM) with Box-Behnken design was employed to simultaneously assess the effects of multiple factors. Finally, optimization of the initial pH and validation of the model were done. The optimum concentrations of the critical components were obtained as follows: 203.96 mL/L of horse serum, 9.64 g/L of glucose, and 9.49 g/L of PPLO broth. The nucleic acid copy number of M. gallisepticum reached 1010.5147 copies/mL and the viable cell count reached a maximum of 109.8451 CCU/mL. This medium reduced the incubation time by approximately 6 h, and M. gallisepticum nucleic acid concentration and viable cell count were higher than those in the modified Frey medium (9.99 and 7 times, respectively). The new liquid medium is likely to improve productivity and reduce the production costs for vaccine-manufacturing companies in the future by reducing incubation times and increasing the growth rate and concentration of M. gallisepticum.

Abstract Objectives To evaluate the efficacy of the O-RADS MRI criteria in the stratification of risk of malignancy of solid or sonographically indeterminate ovarian masses and assess the interobserver agreement of this classification between experienced and inexperienced radiologists. Methods This single-centre retrospective study included patients from 2019 to 2022 with sonographically indeterminate or solid ovarian masses who underwent MRI with a specific protocol for characterisation according to O-RADS MRI specifications. Each study was evaluated using O-RADS lexicon by two radiologists, one with 17 years of experience in gynaecological radiology and another with 4 years of experience in general radiology. Findings were classified as benign, borderline, or malignant according to histology or stability over time. Diagnostic performance and interobserver agreement were assessed. Results A total of 183 patients with US indeterminate or solid adnexal masses were included. Fifty-seven (31%) did not have ovarian masses, classified as O-RADS 1. The diagnostic performance for scores 2–5 was excellent with a sensitivity, specificity, PPV, and NPV of 97.4%, 100%, 96.2%, and 100%, respectively by the experienced radiologist and 96.1%, 92.0%, 93.9%, and 94.8% by the inexperienced radiologist. Interobserver concordance was very high (Kappa index 0.92). Almost all the misclassified cases were due to misinterpretation of the classification similar to reports in the literature. Conclusion The diagnostic performance of O-RADS MRI determined by either experienced or inexperienced radiologists is excellent, facilitating decision-making with high diagnostic accuracy and high reproducibility. Knowledge of this classification and use of assessment tools could avoid frequent errors due to misinterpretation. Critical relevance statement Up to 31% of ovarian masses are considered indeterminate by transvaginal US and 32% of solid lesions considered malignant by transvaginal US are benign. The O-RADs MRI accurately classifies these masses, even when used by inexperienced radiologists, thereby avoiding incorrect surgical approaches. Key points • O-RADS MRI accurately classifies indeterminate and solid ovarian masses by ultrasound. • There is excellent interobserver agreement between experienced and non-experienced radiologists. • O-RADS MRI is a helpful tool to assess clinical decision-making in ovarian tumours. Graphical Abstract

Introduction: Endometriosis is a recurring chronic inflammatory condition that affects females of reproductive age. Endometriosis-associated Pelvic Pain (EAPP) adversely impacts physical, mental, and social well-being. Currently, there is no ideal treatment option. Dienogest, a selective progestin, shows a pronounced effect on endometriosis. Aim: To compare the efficacy and safety of Dienogest at doses of 2 mg and 4 mg/day orally in the treatment of endometriosis. Materials and Methods: An observer-blind, parallel-group, randomised clinical trial was conducted in the Department of Obstetrics and Gynaecology at IPGME&R, SSKM Hospital, Kolkata, West Bengal, India, from April 2016 to December 2017. A total of 190 women aged 20-45 years suffering from endometriosis were recruited and divided into two treatment groups, A and B, receiving daily doses of 2 mg and 4 mg, respectively, for 24 weeks. Relief of EAPP was assessed using the Visual Analogue Scale (VAS) score, and improvement in Quality of Life (QoL) was measured. Treatment-related adverse events were also recorded. Numerical variables were compared using Student’s t-test or the Mann-Whitney U Test, and categorical variables were compared using the Chi-square test or Fisher’s-exact test. Results: The absolute reduction in pelvic pain VAS score was 39.71±8.60 at 24 weeks from the initial score of 70.88 (Mean VAS score before treatment with Dienogest) in Group A, compared to 34.80±6.45 from 69.34 (Mean VAS score before treatment with Dienogest) in Group B (p=0.0001). The difference in mean VAS at 24 weeks between the two groups was statistically significant (p=0.0002). At 24 weeks, 18 (24.66%) patients in Group A experienced an irregular bleeding pattern compared to 27 (40.30%) in Group B, with spotting being the most common issue. Adverse effects such as weight gain, acne, alopecia, depression, and decreased libido were observed in both groups, but they were more pronounced in the 4 mg group. Conclusion: The efficacy and safety results from this dose-ranging study of Dienogest indicate that 2 mg/day is the effective dose for treating endometriosis and offers better tolerability compared to the 4 mg dose.

Abstract Background Ovarian ageing is one of the major issues that impacts female fertility. Mesenchymal stem cell (MSC)-based therapy has made impressive progress in recent years. However, the efficacy and safety of MSCs, as nonautologous components, remain to be further verified. Methods Two common sources of MSCs, umbilical cord-derived MSCs (UC-MSCs) and adipose tissue-derived MSCs (AD-MSCs), were orthotopically transplanted into a mouse model of ovarian ageing to evaluate their therapeutic effects. The safety of the treatment was further evaluated, and RNA sequencing was performed to explore the underlying mechanisms involved. Results After orthotopic transplantation of MSCs into the ovary, the oestrous cycle, ovarian weight, number and proportion of primary follicles, granulosa cell proliferation, and angiogenesis were improved. The effects of AD-MSCs were superior to those of UC-MSCs in several indices, such as post-transplant granulosa cell proliferation, ovarian weight and angiogenesis. Moreover, the tumorigenesis, acute toxicity, immunogenicity and biodistribution of MSCs were evaluated, and both AD-MSCs and UC-MSCs were found to possess high safety profiles. Through RNA sequencing analysis, enhancement of the MAPK cascade was observed, and long-term effects were mainly linked to the activation of immune function. Conclusions Orthotopic transplantation of MSCs displays significant efficacy and high safety for the treatment of ovarian ageing in mice.

Abstract Obesity is associated with increased ovarian inflammation and the establishment of leptin resistance. We presently investigated the role of impaired leptin signalling on transcriptional regulation in granulosa cells (GCs) collected from genetically obese mice. Furthermore, we characterised the association between ovarian leptin signalling, the activation of the NOD-like receptor protein 3 (NLRP3) inflammasome and macrophage infiltration in obese mice. After phenotype characterisation, ovaries were collected from distinct group of animals for protein and mRNA expression analysis: (i) mice subjected to a diet-induced obesity (DIO) protocol, where one group was fed a high-fat diet (HFD) and another a standard chow diet (CD) for durations of 4 or 16 weeks; (ii) mice genetically deficient in the long isoform of the leptin receptor (ObRb; db/db); (iii) mice genetically deficient in leptin (ob/ob); and (iv) mice rendered pharmacologically hyperleptinemic (LEPT). Next, GCs from antral follicles isolated from db/db and ob/ob mice were subjected to transcriptome analysis. Transcriptional analysis revealed opposing profiles in genes associated with steroidogenesis and prostaglandin action between the genetic models, despite the similarities in body weight. Furthermore, we observed no changes in the mRNA and protein levels of NLRP3 inflammasome components in the ovaries of db/db mice or in markers of M1 and M2 macrophage infiltration. This contrasted with the downregulation of NLRP3 inflammasome components and M1 markers in ob/ob and 16-wk HFD-fed mice. We concluded that leptin signalling regulates NLRP3 inflammasome activation and the expression of M1 markers in the ovaries of obese mice in an ObRb-dependent and ObRb-independent manner. Furthermore, we found no changes in the expression of leptin signalling and NLRP3 inflammasome genes in GCs from db/db and ob/ob mice, which was associated with no effects on macrophage infiltration genes, despite the dysregulation of genes associated with steroidogenesis in homozygous obese db/db. Our results suggest that: (i) the crosstalk between leptin signalling, NLRP3 inflammasome and macrophage infiltration takes place in ovarian components other than the GC compartment; and (ii) transcriptional changes in GCs from homozygous obese ob/ob mice suggest structural rearrangement and organisation, whereas in db/db mice the impairment in steroidogenesis and secretory activity.

Abstract The present study aimed to examine the effects of progranulin and omentin on basic ovarian cell functions. For this purpose, we investigated the effects of the addition of progranulin and omentin (0, 0.1, 1, or 10 ng/ml) on the viability, proliferation, apoptosis and steroidogenesis of cultured rabbit ovarian granulosa cells. To determine the importance of the interrelationships between granulosa cells and theca cells, we compared the influence of progranulin and omentin on progesterone and estradiol release in cultured granulosa cells and ovarian fragments containing both granulosa cells and theca cells. Cell viability, proliferation, cytoplasmic apoptosis and release of progesterone and estradiol were measured by Cell Counting Kit-8 (CCK-8), BrdU incorporation, cell death detection, and ELISA. Both progranulin and omentin increased granulosa cell viability and proliferation and decreased apoptosis. Progranulin increased progesterone release by granulosa cells but reduced progesterone output by ovarian fragments. Progranulin decreased estradiol release by granulosa cells but increased it in ovarian fragments. Omentin reduced progesterone release in both models. Omentin reduced estradiol release by granulosa cells but promoted this release in ovarian fragments. The present observations are the first to demonstrate that progranulin and omentin can be direct regulators of basic ovarian cell functions. Furthermore, the differences in the effects of these adipokines on steroidogenesis via granulosa and ovarian fragments indicate that these peptides could target both granulosa and theca cells.

Ovarian mature teratomas are benign, but malignant transformation can occur infrequently, especially in women of advanced age. The tissue that undergoes malignant change is mostly squamous cell carcinoma, although adenocarcinoma has been reported in a small number of cases. The immunostaining results of adenocarcinoma usually show a cytokeratin (CK)7−/CK20+ expression profile, corresponding to lower gastrointestinal tract origin. In this report, we describe a case of mucinous carcinoma arising from an ovarian mature teratoma showing a CK7+/CK20+ profile and discuss its imaging features. A 40-year-old woman presented to her primary care physician with abdominal distension and poor oral intake, and she was referred to our hospital. She had been diagnosed with an ovarian mature teratoma at our institution 3 years earlier. At the current presentation, pelvic magnetic resonance imaging showed a large multilocular cystic mass with adipose tissue extending into the upper abdomen. Densely packed cysts were observed inside the mass, which showed weak contrast enhancement on contrast-enhanced imaging and a mildly high signal on diffusion-weighted imaging. A portion of the cysts also showed abnormal 18F-fluorodeoxyglucose uptake (maximum standardized uptake value, 13.2) on positron emission tomography/computed tomography. The patient was subsequently diagnosed with mucinous carcinoma showing a CK7+/CK20+ profile arising from a mature teratoma by pathologic examination. This mucinous carcinoma arising from a mature teratoma showed a CK7+/CK20+ profile and took the form of densely packed multilocular cysts. In this respect, it was similar to primary ovarian epithelial mucinous carcinoma on both magnetic resonance imaging and pathologic examination despite showing a much higher maximum standardized uptake value than that of primary ovarian mucinous carcinoma. When a large ovarian teratoma contains a large multilocular cyst, the presence of densely packed multilocular cysts should not be missed even in a mass without solid components. Clinicians should consider the possibility of mucinous carcinoma showing a CK7+/CK20+ profile arising from a mature teratoma in such cases.

The potential anti-obesity effects of sea cucumber extract have been reported. However, the individual saponins responsible for these effects are yet to be isolated and characterized. This study aimed to identify the most effective sea cucumber body part for inhibiting lipid accumulation in adipocytes and to elucidate the compounds responsible for this effect using nuclear magnetic resonance (NMR) techniques. Sea cucumber ovary 80 % ethanol extract (SCOE) demonstrated remarkable efficacy in inhibiting adipocyte differentiation compared to other sea cucumber body parts with 50 % or 80 % ethanol extracts. SCOE anti-obesity effect was evaluated in C57BL/6 mice fed a high-fat diet, which revealed significant reductions in body weight, serum lipids, adipose tissue, and liver weight. Using column chromatography, eight saponins were isolated from the SCOE, four of which exhibited potent inhibitory effects on adipocyte differentiation. Of these, three active saponins, holotoxins A, B, and D1, were newly identified. These findings highlight the potential of SCOE and its saponins as effective anti-obesity agents.

Krukenberg tumor is a relatively uncommon metastatic ovarian cancer, typically presenting with abdominal pain and distension, primarily due to bilateral ovarian involvement. Pseudo-Meigs' syndrome, caused by a Krukenberg tumor originating from gastric cancer, is extremely rare. In this study, we report the case of a 39-year-old woman who presented with unusual manifestations of a Krukenberg tumor, where abdominal distension and dyspnea were the primary symptoms. After surgical treatment, a histopathological examination of the ovary revealed the presence of signet ring cell carcinoma. We concluded that this case coincided with Pseudo-Meigs' syndrome. Clinicians should note that Pseudo-Meigs' syndrome should be considered in patients with Krukenberg tumor, ascites, and pleural effusion, as resection of the tumor may provide long-term palliation.

The plasticizer di(2-ethylhexyl) phthalate (DEHP) is known to have endocrine-disrupting properties mediated by its many metabolites that form upon exposure in biological systems. In a previous study, we reported an inverse association between DEHP metabolites in the human ovarian follicular fluid (FF) and the responsiveness of the follicles to controlled ovarian stimulation during in vitro fertilization (IVF) treatments. Here, we explored this association further through molecular analysis of the ovarian FF samples.Ninety-six IVF patients from Swedish (N = 48) and Estonian (N = 48) infertility clinics were selected from the previous cohort (N = 333) based on the molar sum of DEHP metabolites in their FF samples to arrive at “high” (mean 7.7 ± SD 2.3 nM, N = 48) and “low” (0.8 ± 0.4 nM, N = 48) exposure groups. Extracellular miRNA levels and concentrations of 15 steroid hormones were measured across FF samples. In addition, FF somatic cells, available for the Estonian patients, were used for RNA sequencing.Differential expression (DE) and interactions between miRNA and mRNA networks revealed that the expression levels of genes in the cholesterol biosynthesis and steroidogenesis pathways were significantly decreased in the high compared to the low DEHP group. In addition, the DE miRNAs were predicted to target key enzymes within these pathways (FDR

Diet significantly affects reproductive outcomes across species, yet the precise effects of macronutrient compositions beyond caloric intake on reproductive aging are understudied. Existing literature presents conflicting views on the fertility impacts of nutrient-rich versus nutrient-poor developmental diets, underscoring a notable research gap. This study addresses these gaps by examining effects of isocaloric diets with varied protein-to-carbohydrate ratios during both developmental and adult stages on reproductive aging of a large, outbred Drosophila melanogaster population (n = ∼2100). Our results clearly demonstrate an age-dependent dietary impact on reproductive output, initially dominated by the developmental diet, then by a combination of developmental and adult diets in early to mid-life, and ultimately by the adult diet in later life. Importantly, we found that the effects of developmental and adult diets on reproductive output are independent, with no significant interaction. Further investigations into the mechanisms revealed that the effect of developmental diet on fecundity is regulated via ovarioles formation and vitellogenesis; while, the effect of adult diet on fecundity is mostly regulated only via vitellogenesis. These insights resolve disputes in the literature about dietary impacts on fertility and offer valuable perspectives for optimizing fertility strategies in improving public health and conservation efforts in this changing world.

Yaks (Bos grunniens) exhibit exceptional adaptation to the challenging high-altitude environment of the Qinghai-Tibetan plateau, making them the sole bovine species capable of thriving in such exreme conditions. Investigating the cellular and molecular characteristics of yak ovaries across different reproductive states is crucial for gaining insight into their ovarian functions. Herein, the cellular atlases of yak ovaries in different reproductive states were depicted by single-cell RNA-sequencing (scRNA-seq). The cellular atlases of the ovaries were established by identifying specific gene expression patterns of various cell types, including granulosa cells, theca cells, stromal cells, smooth muscle cells, endothelial cells, glial cell, macrophages, natural killer cells, and proliferating cells. The cellular compositions of the ovaries vary among different reproductive states. Furthermore, the granulosa cells comprise six cell subtypes, while theca cells consist of eight cell subtypes. The granulosa cells and theca cells exhibit distinct biological functions throughout different reproductive states. The two cell types were aligned along their respective pseudotime trajectories. Moreover, a cell-to-cell communication network was constructed among distinct cell types within the ovary, spanning the three reproductive states. Notably, during the estrus period, the granulosa cells demonstrated more prominent interactions with other cell types compared to the remaining reproductive states.

ObjectiveTo compare the diagnostic effectiveness of the Gynecologic Imaging Reporting and Data System (GI-RADS) and Neoplasias in the Adnexa (ADNEX) model for the diagnosis of benign and malignant ovarian tumors by junior physicians.MethodsThe sonographic data of 634 patients with ovarian tumors confirmed by pathology in our hospital over 4 years were analyzed retrospectively by junior doctors. The diagnostic efficacy of the GI-RADS and ADNEX models was compared based on pathology.Results(1) Regarding the diagnostic efficacy of the GI-RADS and ADNEX models, the sensitivity was 90.15% and 84.85%, the specificity was 87.65% and 85.86%, the accuracy rates were 88.17% and 85.65%, and the Youden Indices were 0.778 and 0.707, respectively. The areas under the receiver operating characteristic (ROC) curves were 0.924 (95% CI: 0.900-0.943) and 0.933 (95% CI: 0.911-0.951), respectively. The GI-RADS classification was equivalent to that of the ADNEX model in the diagnosis of adnexal tumors (P>0.05). These findings were highly consistent with the pathological results (Kappa values were 0.684 and 0.691, respectively). (2) When differentiating between different pathological types of adnexal tumors, the ADNEX model had the best diagnostic value for distinguishing between benign tumors and stage II-IV ovarian cancer (AUC=0.990, 95% CI: 0.978-0.997).Conclusions(1) The diagnostic efficacy of the GI-RADS and ADNEX models in the diagnosis of benign and malignant ovarian tumors by junior physicians is excellent and comparable. (2) The ADNEX model shows good value for differentiating ovarian tumors of different pathological types by junior physicians.

IntroductionThe reproductive system is tightly regulated by environmental and physiological signals. Melatonin, known as the hormone of darkness, plays a crucial role in regulating both the circadian and reproductive systems in mammals. Hypothyroidism is a key endocrine disorder that harms the reproductive system. Despite many studies on melatonin’s effects on the reproductive system, there is conflicting information regarding melatonin synthesis modulation in hypothyroidism. The objective of this study was to investigate the modulation of plasma melatonin levels and gene expression of Aanat and Asmt in the pineal gland and gonads of rats with hypothyroidism at different times of the day.MethodsFemale and male Wistar rats were divided into control and hypothyroid groups. Hypothyroidism was induced using propylthiouracil (PTU) for 15 days, rats were euthanized six hours after lights on (ZT6), before lights off (ZT11.5), and six hours after lights off (ZT18). Free thyroxine (FT4) and melatonin were quantified in plasma, and gene expressions of melatonin synthesizing enzymes (Aanat and Asmt) were measured in pineal and sexual organs (testis and ovary). Also, morphological analysis was performed in sexual organs.ResultsThe results reveal some disparities between the sexes. Hypothyroidism reduced antral and primary follicles in the ovary, and reduced the weight of testis, epididymis, and prostate. In relation to gene expression, we observed a reduction in Aanat expression in the pineal gland during the light phase (ZT6), and in males, this reduction occurred during the dark phase (ZT18). Regarding Asmt expression, there was a decrease in females also during the dark phase (ZT18). In the gonads, there was an increase in expression in both sexes at ZT11.5. Additionally, it was interesting to observe the association between FT4 levels and Asmt expression in the gonads.ConclusionsThis study showed that acute hypothyroidism can affect components of the melatonergic system in gonads, particularly gene expression of melatonin synthesis enzymes (Aanat and Asmt) contributing to changes in reproduction organs during disease progression. These findings enhance our understanding of melatonin synthesis in the reproductive system during hypothyroidism, showing distinct responses in male and female rats, and suggest that hypothyroidism affects the circadian rhythmicity of melatonin synthesis in a sex-dependent manner.

Betamethasone has been extensively used in medicine in recent years and poses potential hazards to aquatic organisms. This study investigated the reproductive toxic effects of betamethasone exposure in fish, employing female Japanese medaka (Oryzias latipes) as a model. Betamethasone exposure at environmentally relevant concentrations (0, 20, 200, and 2000 ng/L) for a period of 15 weeks resulted in its high accumulation in the ovary, leading to abnormal oogenesis in female Japanese medaka. The production of gonadotropins (LH and FSH) in the pituitary gland was inhibited, and sex steroid biosynthesis in the ovary was significantly influenced at the transcriptional level. The imbalance of androgens and estrogens resulted in a decrease in the E2/T ratio and hepatic VTG synthesis, and the suppression of estrogen receptor signaling was also induced. Furthermore, betamethasone exposure delayed spawning and reduced fertility in the F0 generation, and had detrimental effects on the fertilization rate and hatchability of the F1 generation. Our results showed that environmental betamethasone had the potential to adversely affect female fertility and steroid hormone dynamics in fish.

Ewing sarcoma (ES) is a highly malignant and aggressive small round-cell tumor originating from primitive neuroepithelium and mesenchymal stem cells. It is usually seen in children and adolescents with a male predilection and a preponderance to occur in long bones. Although skeletal/soft tissue ES is encountered in clinical practice, primary ES of the genital tract, particularly bilateral primary ovarian ES, is highly uncommon, with only a handful of cases reported worldwide. Ovarian ES is occasionally reported to involve para-aortic and pelvic lymph nodes in advanced stages. Still, cervical lymph node metastasis from ovarian ES is an infrequent clinical occurrence and, when present, indicates a worse prognosis. Here, we present an intriguing case of bilateral peripheral primary ovarian ES in an adult female, recurring as metastasis in the left submandibular lymph node. This case underlines the importance of keeping metastasis from ES as a possible differential while diagnosing metastatic small round cell tumors in peripheral lymph nodes. It also highlights the usefulness of a minimally invasive diagnostic modality of fine needle aspiration cytology and cell block preparation with applied ancillary techniques of immunohistochemistry and confirmatory molecular testing by fluorescence in-situ hybridization (FISH), for an accurate and quick diagnosis of such entities. The cytological diagnosis of our patient helped in the prompt and early initiation of chemotherapy without requiring any invasive procedure.

Southern stingrays (Hypanus americanus) are relatively large rays that are common and popular in public aquariums because of their size and gentle nature. In aquariums, as well as in the wild, female southern stingrays are fecund. They have a short gestation cycle and can sustain multiple pregnancies each year, each culminating with 2–10 young. This reproductive rate could quickly outpace capacity in managed care and result in a ray surplus. To prevent overpopulation, many aquaria have resorted to single sex groups with a preference for female-only populations. This is an effective way to control population growth, but forces the maintenance of two separated populations of rays; for females this interrupts normal reproductive cycling and replaces it with a protracted non-pregnant condition. An additional consideration is development of reproductive disease in females which is recognized by an enlarged, misshapen, and congested ovary with an abundance of cystic structures and an enlarged uterus with a thickened wall that is often filled with histotroph despite a non-pregnant status. There are no effective long-lasting medical treatments for this type of reproductive disease and mortality is often the result. This report describes a surgical technique for ovariectomy in southern stingrays including outcomes and complications. Ovariectomy as a surgical method prevents unwanted reproduction and has the benefit of reducing reproductive pathologies commonly observed in southern stingrays as they age. Seven stingrays 1–5.2 years old and 42–83.5 cm disc width underwent ovariectomy. After anesthesia, the ovary and a small amount of epigonal was excised via a left para-lumbar incision. Four of the seven rays survived five or more years post-procedure. Two rays died acutely of coelomitis and one ray died of complications unrelated to the procedure. This report details a surgical procedure for ovariectomy in southern stingrays including outcomes, complications, and recommendations.

Introduction: Laying performance is a key factor affecting production efficiency in poultry, but its molecular mechanism is still indistinct. In this study, Yaoshan chickens, a local breed in Guizhou, China, and merchant chickens (GYR) with higher egg yield after the three-line cross improvement hybridization of Yaoshan chickens were used as animal samples.Methods: To explore the regulatory mechanism of the diversities in laying performance, RNA-seq and ultra-performance liquid chromatographytandem mass spectrometry (UPLC—MS/MS) were used to describe the transcriptional and metabolic profiles of the ovaries of Yaoshan and GYR chickens.Results: At the transcriptional level, 288 differentially expressed genes were upregulated in Yaoshan chickens and 353 differentially expressed genes were upregulated in GYR chickens. In addition, GSEA showed that ECM-receptor interactions and the TGF-β signaling pathway were restrained, resulting in increased egg production in GYR chickens. Furthermore, the upregulation of thiamine and carnitine was identified by metabolomic analysis to facilitate the laying performance of hens. Finally, comprehensive analyses of the transcriptome and metabolome found that thiamine and carnitine were negatively correlated with ECM-receptor interactions and the TGF-β signaling pathway, which jointly regulate the laying performance of Yaoshan chickens and GYR chickens.Discussion: Taken together, our research delineates differences in the transcriptional and metabolic profiles of the ovaries of Yaoshan and GYR chickens during the peak egg production period and provides new hypotheses and clues for further research on poultry egg production performance and the improvement of economic benefits.

Female Culter alburnus was faster in growth rate than males. In this study, the gynogenetic G2 and the pseudo-male G2' were used as the female and male parents, respectively, to breed a new national variety “All-female No.1” C. alburnus (AFC). Hormone induction, embryonic development, gonadal differentiation, and growth of AFC were studied. The results showed induction with low concentrations of 17α-methyltestosterone in a indoor-net cage culture was not effective. Under the stimulation of 17α-methyltestosterone, some gonads had a tendency to transform into testis, but not completely. There were three types of gonads in 5-month-old and four types of gonads in 12-month-old fishes, however, they all differentiated into ovaries in 15-month-old fishes. Testosterone propionate and high concentrations of 17α-methyltestosterone in pond culture induction had a good effect resulting in ①a functional pseudo-male with normal testis development that could successfully extrude semen during the breeding period, ②a pseudo-male with normal testis development, but could not extrude semen, and ③the appearance of intersexual glands. The second experiment revealed that with common fish, all-female fish embryo had normal embryonic development. The development time and morphological characteristics of each stage were similar, but the development of the all-female embryo was slightly slower than the common embryos. The gonad differentiation of the all-female embryo were normal and none differentiated into testis, which indicated that all-female could ensure the female sex without affecting the normal gonad differentiation. The correlation between body weight, length, and month-age of all-female and common fish was strong. The all-female had faster growth rate and more uniform growth specification than the common fish. Therefore, the use of testosterone propionate and high concentrations of 17α-methyltestosterone in pond culture induction could avoid complete degeneration of gonads into ovaries. The all-female embryo had the advantages of normal embryonic development and gonadal differentiation, faster growth, and uniform growth specification.

ABSTRACT: Glyphosate (GLY)-based herbicide (GBH) formulations are widely used pesticides in agriculture. The European Union recently decided to extend the use of GLY in Europe until 2034. Previously, we showed that chronic dietary GBH exposure in adult hens resulted in a reversible increase in early mortality in chicken embryos. In this present study, we investigated the GBH effects on metabolism and ovarian functions by using a transcriptomic approach in vivo in young female broilers and in vitro in ovarian explant cultures. We exposed 11-day-old female broilers to 13 mg GLY equivalent/kg body weight/d (GBH13, n = 20), 34 mg GLY equivalent/kg body weight/d (GBH34, n = 20), or a standard diet (control [CT], n = 20) for 25 d. These 2 GBH concentrations correspond to approximatively one-eighth and one-third of the no observed adverse effect level (NOAEL) as defined by European Food Safety Authority in birds. During this period, we evaluated body weight, fattening, food intake, and the weight of organs (including the ovaries). Chronic dietary GBH exposure dose dependently reduced food intake, body weight, and fattening, but increased oxidative stress and relative ovary weight. We analyzed the ovarian gene expression profile in CT, GBH13, and GBH34 broilers with RNA sequencing and showed that differentially expressed genes are mainly enriched in pathways related to cholesterol metabolism, steroidogenesis, and RNA processing. With quantitative polymerase chain reaction and western blotting, we confirmed that GBH decreased ovarian STAR and CYP19A1 messenger RNA and protein expression, respectively. Furthermore, we confirmed that GBH altered steroid production in ovarian explants. We have identified potential regulatory networks associated with GBH. These data provide valuable support for understanding the ovarian transcriptional regulatory mechanism of GBH in growing broilers.

ABSTRACT: This study aims to identify candidate genes related to ovarian development after ovarian tissue transplantation through transcriptome sequencing (RNA-seq) and expression network analyses, as well as to provide a reference for determining the molecular mechanism of improving ovarian development following ovarian tissue transplantation. We collected ovarian tissues from 15 thirty-day-old ducks and split each ovary into 4 equal portions of comparable sizes before orthotopically transplanting them into 2-day-old ducks. Samples were collected on days 0 (untransplanted), 3, 6, and 9. The samples were paraffin sectioned and then subjected to Hematoxylin-Eosin (HE) staining and follicular counting. We extracted RNA from ovarian samples via the Trizol method to construct a transcriptome library, which was then sequenced by the Illumina Novaseq 6000 sequencing platform. The sequencing results were examined for differentially expressed genes (DEG) through gene ontology (GO) function and the Kyoto encyclopedia of genes and genomes (KEGG) pathway analyses, gene set enrichment analysis (GSEA), weighted correlation network analysis (WGCNA), and protein-protein interaction (PPI) networks. Some of the candidate genes were selected for verification using real-time fluorescence quantitative PCR (qRT-PCR). Histological analysis revealed a significant reduction in the number of morphologically normal follicles at 3, 6, and 9 d after ovarian transplantation, along with significantly higher abnormality rates (P < 0.05). The transcriptome analysis results revealed 2,114, 2,224, and 2,257 upregulated DEGs and 2,647, 2,883, and 2,665 downregulated DEGs at 3, 6, and 9 d after ovarian transplantation, respectively. Enrichment analysis revealed the involvement multiple pathways in inflammatory signaling, signal transduction, and cellular processes. Furthermore, WGCNA yielded 13 modules, with 10, 4, and 6 candidate genes mined at 3, 6 and 9 d after ovarian transplantation, respectively. Transcription factor (TF) prediction showed that STAT1 was the most important TF. Finally, the qRT-PCR verification results revealed that 12 candidate genes exhibited an expression trend consistent with sequencing data. In summary, significant differences were observed in the number of follicles in duck ovaries following ovarian transplantation. Candidate genes involved in ovarian vascular remodeling and proliferation were screened using RNA-Seq and WGCNA.