Low egg production in Wanxi white geese limits their commercial viability. This study investigates the role of luteinizing hormone/choriogonadotropin receptor (LHCGR) in regulating granulosa cell function and egg production. Using overexpression, knockdown, single nucleotide polymorphism (SNP) analysis, and dual-luciferase assays, we found that LHCGR is highly expressed in pre-hierarchical follicles of Wanxi white geese, promoting steroidogenesis and granulosa cell proliferation. LHCGR overexpression increases estradiol (E2) and progesterone (P4) levels, whereas knockdown reduces these hormones and increases apoptosis. Three SNPs were identified in the LHCGR promoter, with the wild-type haplotype linked to a 25% increase in egg production. Furthermore, GATA binding protein 2 (GATA2) binds to the wild-type LHCGR promoter, enhancing its expression. These findings demonstrate that the GATA2-LHCGR regulatory network enhances follicular maturation and supports increased egg production in Wanxi white geese, offering targets for genetic selection to enhance egg-laying performance.

Canine mammary tumor is the most common tumor in intact female dogs and poses a growing health burden due to its high malignancy rate and increasing canine populations. However, research on sarcomatous subtypes has been hindered by a lack of representative cell lines. Here, we successfully established a novel canine mammary liposarcoma cell line, designated CMLPS-N1, which represents the first such model derived from a spontaneous tumor. This cell line has been stably maintained for over 80 passages and exhibits an abnormal karyotype, high proliferative and migratory capacity, and strong tumorigenicity in mouse xenografts. Molecular profiling confirmed a phenotype consistent with liposarcoma (MDM2+) and mesenchymal origin (Vimentin+/N-cadherin+), alongside high-risk markers (p53+/Ki67+/Notch1), and hormone receptor expression (ER/PR), while being negative for epithelial (PCK) and HER-2 markers. We used functional assays, including cell proliferation, colony formation, wound healing, and transwell invasion, to confirm its aggressive phenotype. Furthermore, cytotoxicity testing with four chemotherapy agents further supports its utility as a preclinical model for therapeutic screening and mechanistic research. The establishment of CMLPS-N1 enriches the canine mammary tumor cell line repository and provides a valuable experimental model for studying disease mechanisms, developing therapies, and facilitating translational applications.

BackgroundEarly pregnancy loss(EPL) remains a major obstacle to successful in vitro fertilization/intracytoplasmic sperm injection-embryo transfer(IVF/ICSI-ET). Immune dysregulation at the maternal-fetal interface has been increasingly implicated in implantation failure and early miscarriage, while inflammasome-mediated pyroptosis has emerged as a potential contributor to inflammatory tissue injury. However, the direct evidence linking pyroptosis to IVF/ICSI-ET-related EPL remains limited.ObjectiveTo critically appraise direct and indirect evidence regarding immune dysregulation and the potential role of pyroptosis in IVF/ICSI-ET-related EPL, and to define current translational limitations.MethodsA structured review of epidemiological, clinical, and mechanistic studies published between 2020 and 2025 was conducted using literature relevant to EPL, IVF/ICSI-ET, maternal-fetal immune regulation, inflammasome activation, and pyroptosis. Evidence was considered according to its direct relevance to IVF/ICSI-ET-related EPL and interpreted with attention to study design, methodological limitations, and translational applicability.ResultsEstablished clinical predictors of EPL after IVF/ICSI-ET include maternal age, body mass index, endometrial characteristics, and embryonic or sperm-related factors. Immune abnormalities involving uterine natural killer cells, macrophage polarization, and regulatory T-cell imbalance may contribute to impaired maternal-fetal tolerance. Pyroptosis-related pathways, particularly those involving the NLRP3/ASC/Caspase-1 axis, gasdermin D cleavage, and IL-1β/IL-18 release, provide a biologically plausible framework for both physiological epithelial remodeling during implantation and inflammatory injury when dysregulated. Nevertheless, most pyroptosis-related evidence is indirect and derived from pregnancy-related complications or non-reproductive inflammatory diseases rather than from IVF/ICSI-ET-specific EPL studies.ConclusionImmune dysregulation is likely contributes to in IVF/ICSI-ET-related EPL, whereas pyroptosis should currently be regarded as a promising but insufficiently validated mechanistic candidate. Future studies should prioritize cell-specific, temporally resolved, and clinically relevant investigations to clarify whether immune-pyroptosis interactions have diagnostic or therapeutic value in this setting.

Canine mammary tumors are challenging to treat and share multiple molecular and pathological features with human breast cancer, particularly triple-negative breast cancer (TNBC). Demethylzeylasteral (T-96) exhibits significant antitumor activity; however, its mechanism of action against triple-negative breast cancer (TNBC) remains poorly understood. Moreover, pharmacokinetic information regarding intraperitoneal administration remains limited in BALB/c mice. This study aimed to investigate the therapeutic efficacy and underlying mechanisms of T-96 against TNBC and to characterize its pharmacokinetic profile. Transcriptomic analysis revealed that T-96 modulates pathways related to the cell cycle and apoptosis. Consistently, protein-level validation and MDM2 perturbation experiments support the functional involvement of the MDM2–p21 signaling module in T-96-associated phenotypes. At the molecular level, T-96 upregulated the expression of p21, BAX, and cleaved caspase 9 but downregulated the expression of MDM2 and Bcl-2. Pharmacokinetic analysis revealed that T-96 reached its peak plasma concentration (Tmax) at 2.0 h postadministration and had a half-life (t1/2) of 23.5 h. In a mouse breast tumor model, T-96 significantly inhibited TNBC growth, increased the number of necrotic lesions in the tumor center, and reduced liver and lung metastases. Functional validation using transient MDM2 overexpression and inhibitor cotreatment supported a functional contribution of MDM2 to several T-96-associated phenotypes. To our knowledge, this study provides the first evidence that T-96 treatment is associated with reduced MDM2 levels and increased p21 expression, accompanied by G1 phase accumulation and activation of mitochondrial apoptosis markers in TNBC models, collectively contributing to the suppression of TNBC cell growth. Here, MDM2 was identified as a key functional mediator rather than a direct molecular target of T-96. Our findings indicate that the MDM2-p21 signaling module may partially functionally mediate T-96-associated cell cycle arrest and apoptosis, providing preclinical, comparative oncology–relevant evidence to support further evaluation of T-96 in additional models, including those of spontaneously occurring canine mammary tumors.

Canine mammary tumors (CMTs) are the common tumors in female dogs, and approximately 50% of CMTs are malignant tumors, with abnormal regulation of non-coding RNAs being a critical factor in disease progression. Currently, research on long non-coding RNAs (lncRNAs) regulating CMT development remains limited. This study identified a novel lncRNA, aiming to explore the role of lncRNA LOC610012 in CMTs. In this study, immunofluorescence and Western blot analyses were employed to detect protein expression. LncRNA LOC610012 is downregulated in CMT tissues and cells. Stable cells of LOC610012 were constructed by the lentivirus technique. Through a variety of experimental methods, LOC610012 inhibited the proliferation, invasion, and metastasis of CMT cells in in vitro and in vivo experiments conducted using cell culture and mouse models. Mechanistically, LOC610012 regulated the expression of EP3 and GSK-3β by targeting PTGS2, resulting in excessive production of reactive oxygen species (ROS), which inhibited cell viability. Similarly, through transmission electron microscopy, mitochondrial damage caused by LOC610012 was observed in CMT cells, which was manifested as mitochondrial swelling, membrane rupture, and mitochondrial ridge disappearance. PTGS2 could partially restore the inhibition of LOC610012 on cell activity. LOC610012 acts as a tumor suppressor gene in CMTs and as a potential biomarker for the disease.

Ivermectin (IVM) is a macrolide antiparasitic drug, and Metformin (MET) is a biguanide oral hypoglycemic drug. Studies have shown that both of them have obvious anti-tumor effects, but there have been no reports on the combined treatment of Canine breast tumors. This report aimed to investigate the effectiveness and the possible mechanism of drug combination on Canine breast cancers. Mouse breast tumor cells (4T1) and canine breast tumor cells (CMT-1211) were, respectively, treated with IVM, MET, and their combination, and then cell viability was assessed. After that, transcriptomic analysis was performed to study the action pathway of the drug combination with regard to its anti-tumor effects. Reactive oxygen species (ROS) levels were detected by flow cytometry, and autophagosome formation was observed by transmission electron microscopy (TEM). Immunofluorescence detected the cytoplasmic translocation of LC3B and P62 into the nucleus. Western blot detected the protein expressions of LC3B, P62, Beclin1, Bcl-2, p-PI3K, p-AKT, and p-mTOR. Our transcriptomic analysis showed that the combination of IVM and MET regulated the expression of autophagy-related genes and pathways, including the PI3K/AKT/mTOR signaling pathway. Our in vitro experiments showed that the combination of two drugs had a considerably significant effect on cytotoxicity, ROS levels, and the formation of autophagosomes compared to each drug alone. Meanwhile, the in vivo experiments showed that IVM combined with MET had an obvious inhibitory effect on tumor growth in canine breast tumor xenografts. This study concluded that IVM with MET activated autophagy, which killed breast cancer cells by inhibiting the activation of the PI3K/AKT/mTOR pathway and promoting the excessive accumulation of ROS. It offers a theoretical foundation for the synergistic effects of MET and IVM to suppress breast cancer cell activity.

Potassium permanganate (KMnO4) is a commercially available antiseptic used in bovine intrauterine lavage to manage postpartum infections. Lipopolysaccharides (LPS) are well-studied for their ability to induce inflammation and oxidative stress. While KMnO4 is known to cause significant irritation, oxidative stress, and toxicity in uterine tissues, its transcriptional impact and potential for inducing similar molecular damage as LPS have not been fully explored. In this study, we induced oxidative stress in the uterine tissues of Sprague-Dawley (SD) rats using KMnO4 and compared the transcriptional profiles with those treated with LPS. We focused on the differential expression of long noncoding RNAs (lncRNAs) and messenger RNAs (mRNAs) related to oxidative stress, toxicity, and inflammation. RNA sequencing revealed 1125 differentially expressed mRNAs in the KMnO4-treated group and 989 in the LPS-treated group. Additionally, 1649 lncRNAs were differentially expressed in the KMnO4 group compared with 1383 in the LPS group. Gene ontology (GO) and KEGG enrichment analyses showed that 78 pathways were significantly enriched in the KMnO4 group, while 80 pathways were enriched in the LPS group, with 50 pathways shared between the two. This study offers critical insights into the transcriptional profiles associated with KMnO4 exposure and its similarities to LPS-induced damage.

Canine osteosarcoma (OSA) represents a highly aggressive malignancy known for its high rates of recurrence and metastatic potential. This study establishes and characterizes two novel OSA cell lines, OSA-424 and OSA-55, derived from osteoblastic and chondroblastic subtypes, respectively. We conducted a comparative analysis against the established canine chondrosarcoma line Mango. Comprehensive characterization included primary tumor imaging using computed tomography (CT) and magnetic resonance imaging (MRI), as well as histopathological evaluation and immunohistochemical (IHC) profiling. In vitro, functional analyses assessed cellular morphology, karyotypic stability, proliferative capacity, extracellular matrix (ECM) composition, proteomic profiling, migratory potential, invasive behavior, and in vivo tumorigenicity. Both cell lines demonstrated stable propagation beyond 40 passages while maintaining subtype-specific characteristics, with distinct ECM protein expression patterns identified. These validated cellular models provide essential resources for investigating the OSA heterogeneity and advancing the development of subtype-targeted therapeutics.

Interferon-τ (IFN-τ) participates in the establishment of endometrial receptivity in ruminants. However, the precise mechanisms by which IFN-τ establishes bovine endometrial receptivity remain largely unknown. Interferon regulatory factor 1 (IRF1) is a classical interferon-stimulated gene (ISG) induced by type I interferon, including IFN-τ. Leukemia inhibitory factor receptor (LIFR) is a transmembrane receptor for leukemia inhibitory factor (LIF), which is a key factor in regulating embryo implantation in mammals. This study aimed to investigate the roles of IRF1 and LIFR in the regulation of bovine endometrial receptivity by IFN-τ. In vivo, we found IRF1 and LIFR were upregulated in the bovine endometrial luminal epithelium on Day 18 of pregnancy compared to Day 18 of the estrous cycle. In vitro, IFN-τ could upregulate IRF1, LIFR, and endometrial receptivity markers (LIF, HOXA10, ITGAV, and ITGB3) expression, downregulate E-cadherin expression and reduce the quantity of microvilli of bovine endometrial epithelial cells (bEECs). Overexpression of IRF1 had similar effects to IFN-τ on endometrial receptivity, and interference of LIFR could block these effects, suggesting the positive effects of IRF1 on endometrial receptivity were mediated by LIFR. Dual luciferase reporter assay verified that IRF1 could transactivate LIFR transcription by binding to its promoter. In conclusion, IFN-τ can induce IRF1 expression in bovine endometrial epithelial cells, and IRF1 upregulates LIFR expression by binding to LIFR promoter, contributing to the enhancement of bovine endometrial receptivity.

2’-Hydroxychalcone is a hydroxyl derivative of chalcones, which are biosynthetic precursors of flavonoids and rich in the human diet. The anticancer activity of 2’-hydroxychalcone has been reported in several cancers but remains to be investigated in breast cancer. In the current study, 2’-hydroxychalcone showed significant cytotoxicity against breast cancer cell lines MCF-7 and CMT-1211. It could inhibit breast cancer cell proliferation, migration, and invasion in vitro and suppress tumor growth and metastasis in vivo. Mechanistic investigation revealed that the NF-κB pathway was significantly inhibited by 2’-hydroxychalcone treatment accompanied by an excessive intracellular accumulation of reactive oxygen species, induction of endoplasmic reticulum stress, and activation of JNK/MAPK. In addition, 2’-hydroxychalcone elevated the autophagic levels in breast cancer cells equipped with increasing numbers of autophagy vesicles and complete autophagic flux. Finally, autophagy-dependent apoptosis was observed in 2’-hydroxychalcone-induced cell death. In conclusion, 2’-hydroxychalcone enhances the autophagic levels and induces apoptosis in breast cancer cells, which could be contributed to the inhibition of the pro-survival NF-κB signaling, indicating a promising potential for 2’-hydroxychalcone in future anticancer drug development.

In the global progress of bone tumor research, established stable and long-lasting transgenic chondrosarcoma (CSA) cell lines are rare, mainly of murine and human origin, while the establishment of canine CSA cell lines has yet to be reported. This study established a canine CSA cell line to facilitate the basic clinical study of canine CSA. Fifty five cases of canine osteolytic disease were collected, and more than 10 bone tumor samples from dogs with typical clinical signs were used for primary cell culture. A cell line with stable passaging for more than 100 generations and mouse tumorigenic ability was successfully cultured. According to the clinical characteristics of the dog and the histopathological results of the primary tumor, CSA was diagnosed, and the CSA cell line was designated Mango. Immunohistochemical (IHC) results showed that the immunoreactivity of bone gamma-carboxyglutamate protein (BGLAP), secreted protein acidic and rich in cysteine (SPARC), alkaline phosphatase (ALPL), vimentin (VIM) and S100 were positive. However, the immunoreactivity of pan-cytokeratin (PCK), chromogranin A (CGA), and platelet endothelial cell adhesion molecule-1 (CD31) was negative. Immunofluorescence (IF) results showed that the protein expressions in the Mango cell line were consistent with the IHC identification of the primary tumor. The Mango cell line’s doubling time was 43.92 h, and the cell formation rate exceeded 20%. There were abnormal chromosome numbers, hetero staining with toluidine blue, and certain calcification abilities. It could be passaged stably and continuously without changing the cell morphology and characteristics. In vivo, the cells were successfully injected into the nude mice model with a tumorigenic rate of 100%. The immunophenotype of the xenograft tumor was consistent with that of the primary tumor. Therefore, we effectively established a canine CSA cell line. As a promising cell material, this cell line can be used to construct a tumor-bearing model conducive to the subsequent basic research of canine CSA. Moreover, because of its similarity to human CSA, the animal model of CSA is also indispensable for investigating human CSA.

Triple-negative breast cancer (TNBC) escape from immune-mediated destruction was associated with immunosuppressive responses that dampened the activation of tumor-infiltrating CD8 and γδ T cells. TNBC had a higher level of programmed cell death 1-ligand 1 (PD-L1) and indoleamine 2,3-dioxygenase (IDO), compared with other breast cancer subtypes. But, clinical studies have revealed that the response rate of PD-1/PD-L1 antibody for TNBC treatment was relatively low. However, the antitumor responses of human Vγ9Vδ2 T cells or IDO inhibitor in TNBC treatment are unknown. In this study, we found that IDO1 and PD-L1 were highly expressed in TNBC patients. Analysis of the clinical samples demonstrated that Vγ9Vδ2 T cells became exhausted in triple-negative breast cancer patients. And Vγ9Vδ2 T cells combined with αPD-L1 could not further enhance their antitumor responses in vitro and in vivo. However, Vγ9Vδ2 T cells combined with IDO1 inhibitor 1-Methyl-L-tryptophan (1-MT) or Lindrostat showed substantial inhibitory effects on MDA-MB-231 tumor cells. Finally, we found that IDO1 inhibitor promoted T cell’s cytotoxicity by enhancing perforin production. These results converged to suggest the potential application of Vγ9Vδ2 T cells treated with IDO1 inhibitor for TNBC therapy.

It is known that granulosa cells (GCs) mediate gonadotropin-induced oocyte meiosis resumption by releasing EGF-like factors in mammals, however, the detailed molecular mechanisms remain unclear. Here, we demonstrate that luteinizing hormone (LH) surge-induced histone deacetylase 3 (HDAC3) downregulation in GCs is essential for oocyte maturation. Before the LH surge, HDAC3 is highly expressed in GCs. Transcription factors, such as FOXO1, mediate recruitment of HDAC3 to the amphiregulin (Areg) promoter, which suppresses AREG expression. With the LH surge, decreased HDAC3 in GCs enables histone H3K14 acetylation and binding of the SP1 transcription factor to the Areg promoter to initiate AREG transcription and oocyte maturation. Conditional knockout of Hdac3 in granulosa cells in vivo or inhibition of HDAC3 activity in vitro promotes the maturation of oocytes independent of LH. Taking together, HDAC3 in GCs within ovarian follicles acts as a negative regulator of EGF-like growth factor expression before the LH surge.

Polycystic ovary syndrome (PCOS), which is characterized by hyperandrogenism, is a complex endocrinopathy that affects the fertility of 9-18% of reproductive-aged women. However, the exact mechanism of PCOS, especially hyperandrogen-induced anovulation, is largely unknown to date. Physiologically, the natriuretic peptide type C/natriuretic peptide receptor 2 (CNP/NPR2) system is essential for sustaining oocyte meiotic arrest until the preovulatory luteinizing hormone (LH) surge. We therefore hypothesized that the CNP/NPR2 system is also involved in PCOS and contributes to arresting oocyte meiosis and ovulation. Here, based on a dehydroepiandrosterone (DHEA)-induced PCOS-like mouse model, persistent high levels of CNP/NPR2 were detected in anovulation ovaries. Meanwhile, oocytes arrested at the germinal vesicle stage correlated with persistent high levels of androgen and estrogen. We further showed that ovulation failure in these mice could be a result of elevated Nppc/Npr2 gene transcription that was directly increased by androgen (AR) and estrogen (ER) receptor signaling. Consistent with this, anovulation was alleviated by administration of either exogenous human chorionic gonadotropin (hCG) or inhibitors of AR or ER to reduce the level of CNP/NPR2. Additionally, the CNP/NPR2 expression pattern in the anovulated follicles was, to some extent, consistent with the clinical expression in PCOS patients. Therefore, our study highlights the important role an overactive CNP/NPR2 system caused by hyperandrogenism in preventing oocytes from maturation and ovulation in PCOS mice. Our findings provide insight into potential mechanisms responsible for infertility in women with PCOS.

Natriuretic peptides (NPs), brain and C type NPs (BNP and CNP), were involved in the maintenance of porcine oocyte meiotic arrest. The present study investigated the effects of NPs on developmental competence of immature porcine oocytes with follicles of different sizes.

In mammals, oocytes are arrested at the diplotene stage of meiosis I until the pre-ovulatory luteinizing hormone (LH) surge triggers meiotic resumption through the signals in follicular granulosa cells. In this study, we show that the estradiol (E2)-estrogen receptors (ERs) system in follicular granulosa cells has a dominant role in controlling oocyte meiotic resumption in mammals. We found that the expression of ERs was controlled by gonadotropins under physiological conditions. E2-ERs system was functional in maintaining oocyte meiotic arrest by regulating the expression of natriuretic peptide C and natriuretic peptide receptor 2 (NPPC/NPR2), which was achieved through binding to the promoter regions of Nppc and Npr2 genes directly. In ER knockout mice, meiotic arrest was not sustained by E2 in most cumulus–oocyte complexes in vitro and meiosis resumed precociously in pre-ovulatory follicles in vivo . In human granulosa cells, similar conclusions are reached that ER levels were controlled by gonadotropins and E2-ERs regulated the expression of NPPC/NPR2 levels. In addition, our results revealed that the different regulating patterns of follicle-stimulating hormone and LH on ER levels in vivo versus in vitro determined their distinct actions on oocyte maturation. Taken together, these findings suggest a critical role of E2-ERs system during oocyte meiotic progression and may propose a novel approach for oocyte in vitro maturation treatment in clinical practice.

Interferon-τ (IFN-τ) signals pregnancy recognition in ruminants. We investigated the effects of IFN-τ produced by embryo trophoblastic cells (ETCs) on expression of bovine leukocyte antigen-I (BoLA-I), a bovine analogue of human MHC-I, in endometrial luminal epithelial cells (EECs) during early pregnancy in dairy cows. Expression of IFN-τ and BoLA-I was increased in endometrial tissues during early pregnancy. Expression of the anti-inflammatory cytokine IL-10 was increased in endometrial tissues, while expression of the pro-inflammatory cytokine IL-6 was decreased, indicating immunosuppression. Progesterone increased IFN-τ expression in EECs. IFN-τ increased p-STAT1 and p-STAT3 levels in EECs, but reduced TRAF3 levels. In addition, IFN-τ increased expression of BoLA-I and IL-10, but decreased expression of IL-6 in EECs. These results indicate that IFN-τ enables stable implantation in dairy cows by increasing expression of BoLA-I, and by immunosuppression mediated by increased IL-10 and decreased IL-6 expression.

Recent studies have shown that C‐type natriuretic peptide (CNP) serves as a key control system during mouse oocyte maturation. We used pig models (in vitro and in vivo) to explore the role played by the natriuretic peptide family in porcine oocyte maturation. We reported the expression and location of natriuretic peptide system in different stages of porcine antral follicles. Atrial natriuretic peptide (ANP) and CNP were expressed primarily in granulosa cells, whereas brain natriuretic peptide (BNP) and natriuretic peptide receptor‐B (NPRB) receptor were expressed in granulosa cells (both cumulus and mural granulosa cells) and thecal internal cells, and the natriuretic peptide receptor‐A (NPRA) receptor predominantly in thecal cells. Upon in vitro culture, BNP and CNP maintained meiotic arrest of oocytes associated with cumulus cells. The expression levels of BNP, CNP, and the NPRB receptor increased upon treatment of prepubertal gilts with pregnant mare's serum gonadotropin and decreased upon subsequent human chorionic gonadotropin injection. Such dynamic changes in the expression of natriuretic peptides and their receptor paralleled the proportions of oocytes exhibiting nuclear maturation in vivo. These data indicated that BNP and CNP co‐contributed to maintaining porcine meiotic arrest under physiological condition and lutenizing hormone (LH) relieved this inhibitory effect by decreasing the expression levels of BNP and CNP in vivo. Our present work, combined with previous data, improved the understanding of the oocyte meiotic arrest mechanisms and further revealed that natriuretic peptides serve as oocyte maturation inhibitor (OMI) to inhibit oocyte maturation in mammals. J. Cell. Physiol. 230: 71–81, 2015. © 2014 Wiley Periodicals, Inc.