Testosterone synthesis in Leydig cells requires precise coordination between LH signaling and cholesterol metabolism, but the mechanisms underlying this regulation remain incompletely understood. In this study, we reveal a previously unrecognized YAP1-ACSL4 axis essential for LH-induced testosterone production by integrating transcriptomic profiling and functional studies in Leydig cells. Transcriptomic sequencing identified ACSL4 and YAP1 as key regulators of testosterone synthesis induced by LH. The inhibition of YAP1 led to the downregulation of ACSL4 expression, which significantly reduced testosterone production. Similarly, si-ACSL4 directly reduced testosterone production in Leydig cells. Notably, transcriptomic sequencing after interfering with ACSL4 showed that SOAT1 is a key molecule downstream of ACSL4. SOAT1 promoted testosterone synthesis by inhibiting esterification of free cholesterol and subsequently increasing cholesterol availability providing materials for testosterone synthesis. Mechanistically, ACSL4 modulates cholesterol availability through SOAT1 suppression, shifting metabolic flux from cholesterol ester storage toward cholesterol. These results indicate that LH promotes testosterone synthesis by upregulating YAP1 expression, which in turn increases ACSL4 expression, inhibits SOAT1 expression, and raises the concentration of free cholesterol. This finding provides the first evidence linking ACSL4-mediated lipid metabolism to gonadotropin-regulated steroidogenesis.

Fusarium root rot on ginseng is an important root disease that seriously affects ginseng (Panax ginseng) yield and quality. However, the species categories and distribution of Fusarium causing ginseng root rot in China have not been systematically examined. A total of 571 pure Fusarium isolates were obtained from 2018 to 2019 from 14 ginseng-producing regions in Jilin, Liaoning, and Heilongjiang provinces, China. Based on multilocus sequence analysis of ITS-tef1-rpb2 and morphological characteristics, the 571 Fusarium isolates were identified as F. oxysporum (accounting for 47.46% of the total isolates), F. solani (35.38%), F. equiseti (5.78%), F. proliferatum (2.80%), F. cerealis (1.75%), F. semitectum (1.75%), F. acuminatum (1.75%), F. redolens (1.58%), F. verticillioides (1.05%), and F. graminearum (0.70%). Among them, F. oxysporum and F. solani were the dominant species, and F. graminearum and F. verticillioides were first recorded on ginseng in China. The fungicides captan and hymexazol were tested in vitro for their inhibitory activities against 10 Fusarium species. All Fusarium species displayed enhanced sensitivity to captan compared with hymexazol. The protective effects of captan against ginseng root rot caused by F. oxysporum ranged from 94.07 to 97.78% at concentrations of 200, 400, and 600 μg·ml-1; however, its curative effects were much lower, ranging from 29.63 to 35.56%. In comparison, hymexazol exhibited protective effects of only 24.00 to 52.00% and curative effects of 16.00 to 45.33% at concentrations of 600 to 1,000 μg·ml-1. In addition, microconidia were highly sensitive to both fungicides, compared with mycelia and macroconidia.

The unresolved mechanisms underlying heterosis in beef cattle production traits hinder the optimization of crossbreeding strategies. In this study, we generated Wagyu × Hereford hybrid cattle (F1) and observed significantly enhanced growth rates compared to purebred Hereford controls, prompting further investigation into the epigenetic regulatory basis of heterosis in bovine muscle development. Combined microRNA-seq and RNA-seq profiling of the longissimus dorsi muscle identified 17 differentially expressed miRNAs and 489 differentially expressed genes (DEGs), enabling the construction of a comprehensive miRNA-mRNA regulatory network. Notably, decreased expression of bta-novel-miR-357 (novel-miR-357) and increased expression of S100A2 (S100 Calcium-Binding Protein A2) mRNA were observed in the hybrid cattle. Functional assays in primary bovine myoblasts demonstrated that novel-miR-357 and siS100A2 inhibited cell proliferation and promoted apoptosis. Dual-luciferase reporter assays confirmed that novel-miR-357 directly binds to the 3'UTR of S100A2 mRNA, establishing a targeted regulatory relationship. Furthermore, novel-miR-357 was shown to inhibit proliferation and promote apoptosis of bovine myoblasts through targeting S100A2 mRNA. These findings reveal a novel epigenetic regulatory axis (novel-miR-357/S100A2 mRNA) involved in early muscle development and provide an important molecular mechanism for deeper understanding of the epigenetic regulatory processes underlying heterosis in cattle. Nevertheless, given the polygenic complexity of heterosis, it is important to recognize that this pathway likely represents only one of several contributing factors to this complex trait.

The follicle was regarded as the fundamental functional unit of the ovary, and dysfunction of its granulosa cells was known to impair follicular development and delay ovulation. In bovine, reproductive‑tract infections frequently elicited excessive inflammatory responses that resulted in follicular dysfunction, a process largely mediated by lipopolysaccharide (LPS) induced programmed death of ovarian granulosa cells. Necroptosis was shown to contribute to the regulation of follicular development while simultaneously promoting the release of pro‑inflammatory mediators. Although α‑ketoglutarate acid (α‑KG) was recognized as an anti‑inflammatory and antioxidant metabolite, its capacity to attenuate LPS‑induced necroptosis in bovine granulosa cells-and the underlying. To investigate the protective mechanism of α-KG against LPS-induced functional impairment in ovarian granulosa cells, primary granulosa cells were isolated from bovine follicles and cultured in vitro. A series of assays-including qRT-PCR, Western blotting, flow cytometry, immunofluorescence staining, and transmission electron microscopy-were conducted to assess necroptosis, autophagy, and mitochondrial function. LPS was found to induce necroptosis in bovine ovarian granulosa cells by promoting autophagy. This process was accompanied by mitochondrial dysfunction and disruption of mitochondrial cristae structure. Notably, α-KG was shown to inhibit autophagy-mediated necroptosis by restoring mitochondrial membrane potential, decreasing intracellular levels of ROS, Ca²⁺, and thereby attenuating inflammatory responses. In summary, the findings indicated that α-KG attenuated mitochondrial dysfunction-associated autophagy, necroptosis, and inflammatory responses in bovine ovarian granulosa cells following LPS exposure in vitro. These results suggested that α-KG may potentially provide a pharmacological basis for mitigating inflammation-associated follicular dysplasia in bovine, although further in vivo studies are required to confirm these effects.

Blastocyst cryopreservation is a challenging process due to ice crystallization induced by blastocoel fluid. While aquaporin-3 (AQP3) overexpression improves cryotolerance, there is a lack of pharmacological regulators of AQP3 that can be easily applied in clinical settings. Notably the antidiabetic drug pioglitazone (PIO), despite its association with edema in patients, has been shown to potentially induce cellular dehydration through the modulation of AQP3. Here, we have demonstrated that PIO improves vitrification outcomes by orchestrating AQP3-mediated fluid expulsion and bolstering antioxidant defense mechanisms. Treatment of mouse blastocysts with 2 μM PIO in a hyperosmotic solution resulted in a notable contraction of blastocysts (reduced by 25.95 %, P < 0.01), correlating with 2.6-fold AQP3 up-regulation, compared to the control. This rapid dehydration yielded 95 ± 3.60 % post-thaw survival (vs. 82.5 ± 3.51 % Control, P < 0.001), while also reducing levels of Reactive oxygen species (ROS, P < 0.05) and Dihydroethidium (DHE) (P < 0.05) and increasing levels of glutathione (GSH, P < 0.05) and mitochondrial membrane potential (ΔΨm, P < 0.05) significantly. Mechanistically, PIO reduced DNA damage (γ-H2AX, P < 0.01) while increasing P53 expression (P < 0.01). Crucially, these effects were conserved in bovine Blastocysts, at the concentration of 4 μM, achieving 88.65 ± 8.31 % survival with PIO compared to 82.23 ± 7.48 % in the control group, highlighting the cross-species applicability of the findings Our findings position PIO as a dual-functional cryoadjuvant by coordinating rapid water efflux through AQP3 with antioxidant defense mechanisms. This repurposing approach, leveraging the anti-diabetic agent PIO as a cryo-adjuvant, offers an immediately implementable optimization pathway for the standardized enhancement of animal embryos vitrification protocols.

The cryopreservation technique for sperm is a widely recognized approach utilized in artificial insemination. Previous studies have demonstrated that sperm quality is influenced by ice crystal formation and oxidative stress during the freezing process. Moreover, the time of cryostorage also has an impact on sperm quality. Nevertheless, there is a scarcity of comprehensive research concerning the years of storage on bovine sperm. In this study, bovine sperm samples cryopreserved for 1, 5, and 7 years were analyzed for motility, functionality membrane integrity, oxidative stress markers, apoptosis, DNA fragmentation, and fertilizing ability. Post-thaw assessments revealed a significant decline in antioxidant capacity and an increase in lipid peroxidation and apoptosis in the 7-year group compared to the 1-year group (P < 0.05). Despite relatively stable sperm membrane and acrosome integrity, DNA damage and oxidative stress were elevated over time. Notably, while cleavage rates remained unchanged, blastocyst development was significantly reduced after 7 years of storage (P < 0.05). These findings suggest that prolonged cryopreservation negatively affects sperm function and embryo development, emphasizing the need to consider storage duration in reproductive applications.

Endometrial injury triggers inflammation responses, and persistent inflammation is a recognized cause of infertility. Emerging evidence underscores the critical role of the vagus nerve in modulating immunity. Although vagotomy is known to induce systemic inflammation, the specific mechanism by which the vagus nerve regulates uterine health is still unclear. In this study, we employed a vagotomy model to investigate the therapeutic potential of pantothenic acid in alleviating endometrial injury. Our results showed that left cervical vagotomy reduces the integrity of the endometrium and the expression of barrier proteins such as Claudin-3, Occludin, and ZO-1. Vagotomy increases the levels of inflammatory cytokines (TNF-α and IL-1β) and LPS in uterine tissue and serum through the NF-κB signaling pathway. Furthermore, we found that vagotomy promotes ferroptosis by decreasing the protein expression of SLC7A11 and GPX4 and increasing the level of COX2. Vagotomy significantly altered thecompositionof the uterine microbiota,characterized by a significant enrichment of Rodentibacter and a depletion of Vagococcus and Acetobacter. Next, we found that vagotomy can cause an increase in serum levels of lysoPE 20:4, antipyrine, and lysoPE18:2, as well as a decrease in levels of hexanoyl-L-Carnitine and pantothenic acid. When pantothenic acid was supplemented, the endometrial injury caused by vagotomy was reversed. Pantothenic acid increased the expression of barrier proteins in the endometrium and reduced the content of inflammatory cytokines in uterine tissue and serum of mice. At the same time, pantothenic acid also reversed the degree of ferroptosis induced by vagotomy in uterine tissue and serum. Our study demonstrates that vagotomy disrupts the endometrial microbiota and promotes endometrium injury and the markers of ferroptosis via the NF-κB pathway. Pantothenic acid supplementation alleviates vagus nerve-mediated endometrial injury. These results highlight that vagus nerve regulation of uterine health through pantothenic acid is a promising strategy.

Nickel, a heavy metal with industrial applications and as a feed additive for livestock, can adversely impact reproductive function and gamete quality when present in excessive amounts in the animal feed environment. In this study, the results indicate that nickel exposure hampers polar body extrusion and cumulus cell expansion, thereby diminishing oocyte quality and developmental competence. Furthermore, nickel exposure reduces glutathione (GSH) levels in oocytes, leading to excessive accumulation of reactive oxygen species (ROS), provoking oxidative stress and mitochondrial impairment. This cascade initiates mitochondrial autophagy, upregulates the expression of autophagy-related proteins Parkin and PINK1, promotes LC3 binding to autophagosomes. Nickel exposure disrupts calcium homeostasis and induces endoplasmic reticulum stress (ERS). Inhibiting ERS effectively alleviates the deterioration of oocytes quality caused by nickel exposure and inhibits mitochondrial autophagy. Proteomics further confirms nickel’s exposure detrimental effects on mitochondria and ER, impacting cellular processes. Employing bovine oocytes as a model, consistent phenotypes were observed. These results indicate that nickel exposure disrupts intracellular calcium homeostasis, elicits ERS, disrupts cellular calcium homeostasis, diminishes the quality of pig oocytes, impairs oocyte maturation and developmental potential, and instigates mitochondrial autophagy.

Sperm freezability exhibits marked individual variability, yet the mechanisms remain unclear. Using bulls as the experimental model, we integrated proteomic (sperm) and metabolomic (seminal plasma) analyses of high-freezability (HF) and control (CF) bulls to identify key biomarkers associated with sperm freezability. Post-thaw motility and membrane integrity were significantly higher in HF bulls (p < 0.05). Sperm proteome analysis revealed upregulated antioxidant proteins (PRDX2, GSTM4), heat shock proteins (HSP70, HSP90), and key enzymes in arginine and proline metabolism (PRODH, LAP3). Seminal plasma metabolomics revealed elevated spermine in HF bulls. Meanwhile, we found that spermine abundance was positively correlated with post-thaw motility, as well as with the expression levels of both PRODH and LAP3 (r > 0.6, p < 0.05). Functional validation demonstrated that 200 μM spermine supplementation in cryopreservation extenders enhanced post-thaw motility, kinematic parameters (VAP, VSL, VCL), membrane integrity, and acrosome integrity (p < 0.05). Concurrently, spermine enhanced antioxidant enzyme (SOD, CAT, GSH-Px) activity and reduced ROS and MDA levels (p < 0.05). Our study reveals a spermine-driven antioxidant network coordinating sperm–seminal plasma synergy during cryopreservation, offering novel strategies for semen freezing optimization.

BACKGROUND:Endometritis is a highly prevalent reproductive disorder in cows, causing serious adverse effects on reproductive performance, which brings huge economic losses to the livestock industry. Staphylococcus aureus is detected in a high proportion of endometritis pathogens (alone or in combinations of infections). Uterine microbial composition plays an important role in endometritis. OBJECT AND METHOD:In order to determine the role of S. aureus in endometritis, we established an endometritis model using this bacterium and utilized metagenomics to detect the structure and function of the bovine uterine microbiota. RESULTS:We found that S. aureus infection significantly increased the relative abundance of bacteria such as Escherichia coli, Trueperella pyogenes, and Streptococcus spp., while reducing the relative abundance of Akkermansia and Prevotella bacteria. The functions of microorganisms in the uterus are mainly manifested in metabolic levels, including carbohydrate metabolism, amino acid metabolism, energy metabolism, and lipid metabolism processes. The number of genes continues to increase with the duration of S. aureus infection, which disrupts the balance that maintains the bovine uterine flora. CONCLUSION:This study provides a descriptive analysis of changes in the uterine microbiota of cows infected with S. aureus, which contributes to a new understanding of uncultured or unidentified pathogenic bacteria.

The cattle industry is critical for agriculture's development, and reproductive technology has revolutionized its development over the past century. The core task of developing the beef cattle industry is to increase quantity and improve quality, which cannot be separated from the key link of reproduction. It involves artificial insemination (AI), semen collection and freezing, multiple ovulation embryo transfer (MOET), and genome editing. Each technology greatly contributed to the industry's development at the time. In this review, we systematically summarize the development process of beef cattle breeding technology and provide reasonable suggestions for the needs of enterprises and individual breeders of different scales. Additionally, we summarize the contributions of these technologies to industry development and their impact on cattle farming today. Scientific feeding methods combined with current reproductive technology can fully realize the production potential of beef cattle and improve economic benefits, which has positive significance for expanding the beef cattle industry scale and promoting sustainable cattle industry development.

The cellular basis of testicular development and spermatogenesis for the extreme sperm density in chickens (100-fold higher than mammals) remains poorly defined. A comprehensive understanding of the molecular characteristics driving poultry testicular development is crucial for explaining this enhanced spermatogenic capacity. Here, we first established a single-cell transcriptome profile of chicken testes from hatching to maturity, identifying the dynamic transcriptional characteristics of germ cell fate transition and exploring the developmental characteristics of Sertoli cells and Leydig cells. Multi-species comparisons revealed a higher proportion of germ cells and the unique adaptations of Sertoli cells in chicken testes. Most importantly, our results demonstrated that Sertoli cells dominated in somatic composition of mature chicken testes, and proliferating Sertoli cells persisted in chicken testes even after sexual maturity, while no proliferating Sertoli cells in mammals. We also found a richer interaction network between chicken testicular cells, especially the specific activation of Sertoli cell interaction signals, such as TGF-β, BMP, EGF, and activin. These adaptations of Sertoli cells may support the spermatogenic superiority in chickens. Additionally, our results indicated that cAMP responsive element binding protein 5 (CREB5) played a crucial role in maintaining the maturation and function of chicken Sertoli cells, and circadian rhythm promoted testosterone secretion and the development of Leydig cells. Our study revealed that the sustained proliferative capacity of Sertoli cells, their enriched signaling network, and the regulatory roles of CREB5 and circadian rhythms collectively represented unique testicular adaptations in chickens. These findings may hold extraordinary significance in understanding the molecular characteristics of poultry testicular development, and provide a plausible framework for explaining enhanced spermatogenesis in poultry.

The necroptosis of granulosa cells has been proven to be one of the important triggers of follicular atresia, which is an important cause of reduced reproductive capacity in cows. The rapid growth of granulosa cells is accompanied by endoplasmic reticulum stress (ERS), leading to granulosa cell death. However, the link between ERS and necroptosis, as well as its mechanism in bovine granulosa cells is still unclear. Itaconic acid is an endogenous anti-inflammatory and antioxidant small-molecule compound that can alleviate ERS. Therefore, the aim of the current study is to evaluate the effect of ERS on necroptosis and investigate the ameliorative effect of itaconic acid against ERS-induced necroptosis in granulosa cells. Bovine granulosa cells were treated with tunicamycin (Tm) to induce ERS. After the addition of the necroptosis inhibitor Nec-1 and the detection of the necroptosis inducer acetylcholinesterase (AChE), flow cytometry, transmission electron microscopy, and mass spectrometry were used to analyze the expression of itaconic acid and IRG1 in the granulosa cells. In addition, the role of the PERK pathway downstream of ERS in ERS-induced necroptosis was also investigated. We report here that ERS can induce necroptosis in granulosa cells. Itaconic acid supplementation significantly attenuates the effect of ERS-induced damage. In summary, this research provides a scientific basis and a drug reference for treating follicular atresia and improving bovine reproductive capacity.

Mercury, a prevalent heavy metal, negatively impacts oocyte maturation. However, the exact mechanism by which methylmercury chloride (MMC) affects this process remains elusive. The present study found that MMC administration triggered meiotic failure in oocytes by disrupting cumulus cell expansion, leading to compromised spindle apparatus and altered chromosomal architecture, which are crucial for oocyte development. This disruption is characterized by abnormal microtubule organization and defective chromosome alignment. Additionally, MMC exposure caused oxidative stress-induced apoptosis due to mitochondrial dysfunction, as indicated by decreased mitochondrial membrane potential, mitochondrial content, mitochondrial DNA copy number, and adenosine triphosphate levels. Proteomic analysis identified 97 differentially expressed proteins, including P62, an autophagy marker. Our results confirmed that MMC induced autophagy, particularly through the hyperactivation of the mitochondrial autophagy to remove damaged and normal mitochondria. The mitochondrial reactive oxygen species (ROS) scavenger Mito-TEMPO alleviated oxidative stress and mitochondrial autophagy levels, suggesting that mitochondrial ROS initiates this autophagic response. Notably, MMC activates mitochondrial autophagy via the monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signal pathway due to mitochondrial dysfunction. In vivo studies in mice revealed that MMC exposure decreased reproductive performance, attributed to excessive mitochondrial autophagy leading to reduced oocyte quality. Overall, these findings demonstrate that MMC exposure impairs oocyte maturation via the hyperactivation of mitochondrial autophagy induced by mitochondrial dysfunction.

Accumulation of ketone bodies in the blood or tissues can trigger ketosis, exerting detrimental effects on bovine oocytes maturation. Exposure to its primary component, (3- hydroxybutyric acid ((3HB), disrupts mitochondrial function, culminating in the excessive buildup of reactive oxygen species (ROS) and subsequent initiation of apoptosis in oocytes. These ultimately result in poor oocyte quality. Melatonin, recognized for its endogenous antioxidant properties, is capable of mitigating ROS levels and enhancing the expression of antioxidant enzymes. In this study, we explored the protective effects of melatonin on the damages induced by (3 HB. Melatonin was added at a concentration of 10-9 M to the culture medium on bovine oocytes. Parameters including first polar body extrusion rate, mitochondrial membrane potential, ROS, cell apoptosis were assessed. Results showed that melatonin could restore bovine oocyte maturation rate, enhance mitochondrial function, reduce cell apoptosis rate, and mitigate oxidative stress levels. Notably, Nrf2 signaling pathway inhibitor ML385 significantly attenuated the protective effects of melatonin on oxidative stress induced by (3 HB exposure. In summary, our study demonstrates that melatonin can protect oocytes from oxidative stress induced by (3 HB exposure, with indications that this protective mechanism may be mediated through the Nrf2 pathway.

Furosemide sodium, a loop diuretic that inhibits the Na-K-2Cl cotransporter (NKCC1), is commonly used in clinical treatment of edema. In this study, we investigated whether furosemide sodium could improve the survival and quality of vitrified mouse and bovine blastocysts by promoting the expulsion of blastocoelic fluid, thus enhancing cryoprotectant penetration and reducing ice crystal formation. Mouse and bovine blastocysts were vitrified using equilibration solutions (ES) and vitrification solutions (VS) with or without furosemide sodium, then thawed for survival and quality assessment. Results indicated that furosemide sodium significantly improved post-thaw survival rates (P < 0.05). It also markedly reduced reactive oxygen species (ROS) levels (P < 0.05). Additionally, furosemide sodium increased glutathione (GSH) levels (P < 0.05) and enhanced mitochondrial function. The improvement in mitochondrial function was reflected in increased mitochondrial membrane potential, which are key indicators of mitochondrial health and energy status (P < 0.05). Immunofluorescence analysis revealed that furosemide sodium downregulated NKCC1 protein levels and upregulated Aquaporin 3 (AQP3) protein levels (P < 0.05), indicating that furosemide sodium modulates ion transport and water regulation. This modulation likely contributes to the closure of sodium-potassium ion channels, reducing ion influx and promoting water efflux from the blastocyst. This closure limited extracellular sodium and potassium influx, significantly reducing these ions within the blastocyst cavity (P < 0.05). Similar effects were observed in both mouse and bovine blastocysts, where furosemide sodium promoted fluid expulsion from the blastocoel, thereby enhancing vitrification efficiency. These findings suggested that furosemide sodium may be a promising agent to improve vitrification efficiency in blastocysts by increasing osmotic pressure differences across the blastocoel.

Epigenetics studies heritable changes in gene expression without altering the DNA sequence and is involved in diverse biological processes. In male reproduction, Leydig cells, the main site of testosterone synthesis, play an important role in maintaining the reproductive process. However, the role of epigenetics in the mechanism of testosterone synthesis in Leydig cells is still not well understood. This review systematically describes how classic epigenetic modifications such as DNA methylation, RNA methylation, histone modification, and non-coding RNA regulate the testosterone synthesis process of Leydig cells in different species. Accumulating evidences revealed that epigenetics can regulate the process of testosterone synthesis in Leydig cells. In future, we aim to provide new ideas for male reproduction by investigating the relationship between testosterone synthesis mechanisms and epigenetics.

Maternal obesity impairs placental angiogenesis, increasing the risks of gestational metabolic disorders and fetal developmental compromise. Odd-chain saturated fatty acids (OCS-FAs), particularly pentadecanoic acid (PA, C15:0), demonstrate protective metabolic properties, yet their roles in placental angiogenesis remain unexplored. In this study, pregnant mice were fed a high-fat diet (HFD, 60% kcal) or isocaloric HFD supplemented with 1% (w/w) PA ethyl-ester (HFD + PA). Porcine iliac artery endothelial cells (PIEC) were used to assess angiogenic mechanisms in vitro. Our results showed that PA supplementation induced mild glucose intolerance (elevated 90/120 min glycemia, P < 0.05) without altering the body composition, fasting insulin, HOMA-IR, or plasma lipids (TG, T-CHO, HDL/LDL). PA augmented labyrinth zone (LZ) vascularization (P = 0.052) and upregulated fetal T-CHO transport (P = 0.002). RNA-seq revealed PA-activated PI3K-AKT signaling and enhanced the pro-angiogenic factor expression (HIFα, Lrp1, Flt4, MMP2/14, P < 0.05). Immunohistochemistry confirmed PI3K activation and increased CD31 endothelium in LZ. PA promoted PIEC tube formation (12.5 µM, P < 0.01) in vitro, while heptadecanoic acid (HA, C17:0) had no such effect and inhibited PIEC (12.5-25 µM, P < 0.01). PA's pro-angiogenic effect was abolished by PI3K inhibitor 3-MA (P < 0.01). In conclusion, PA promotes placental angiogenesis by activating the placental PI3K-AKT signaling, despite mild maternal glucose intolerance. These findings highlight PA's potential as a functional nutrient for mitigating gestational metabolic complications.

Contrastive Language-Image Pre-training (CLIP) has achieved success on multiple downstream tasks by aligning image and text modalities. However, the nature of global contrastive learning limits CLIP's ability to comprehend compositional concepts, such as relations and attributes. Although recent studies employ global hard negative samples to improve compositional understanding, these methods significantly compromise the model's inherent general capabilities by forcibly distancing textual negative samples from images in the embedding space. To overcome this limitation, we introduce a Decoupled Global-Local Alignment (DeGLA) framework that improves compositional understanding while substantially mitigating losses in general capabilities. To optimize the retention of the model's inherent capabilities, we incorporate a self-distillation mechanism within the global alignment process, aligning the learnable image-text encoder with a frozen teacher model derived from an exponential moving average. Under the constraint of self-distillation, it effectively mitigates the catastrophic forgetting of pretrained knowledge during fine-tuning. To improve compositional understanding, we first leverage the in-context learning capability of Large Language Models (LLMs) to construct about 2M highquality negative captions across five types. Subsequently, we propose the Image-Grounded Contrast (IGC) loss and Text-Grounded Contrast (TGC) loss to enhance vision-language compositionally. Experimental results across both general and compositional reasoning tasks validate the effectiveness of the DeGLA framework. Our code is released at https://github.com/xiaoxing2001/DeGLA.

Recently, evidence has indicated that mastitis is closely associated with the ruminal dysbiosis caused by subacute ruminal acidosis (SARA) and subsequent low-grade endotoxemia (LGE). However, whether and how ruminal metabolic dysbiosis influences SARA-associated mastitis is still unclear. Using untargeted metabolomics, we found that cows with SARA-associated mastitis exhibited altered ruminal metabolic profiles, particularly a reduced level of phytosphingosine (PS), compared to healthy cows. Oral administration of PS to mice alleviated LGE-induced mastitis, as evidenced by attenuated mammary injury and improved function of mammary tight junctions (TJs). Furthermore, we demonstrated that LGE induced significant gut dysbiosis, characterized by increased abundances of opportunistic pathogens such as Enterococcus, which was mitigated by PS treatment. Interestingly, transplantation of both fecal microbiota (FMT) and matched sterile supernatant (FST) from PS-treated mice alleviated LGE-induced mastitis, restored the blood-milk barrier, and modulated the gut microbiota in recipient mice following LGE exposure. Additionally, PS and PS-FMT treatments increased the abundances of fecal short-chain fatty acid (SCFA) producers, accompanied by elevated fecal SCFA levels, particularly butyric acid, in PS- and PS-FMT-treated mice. Butyric acid was negatively correlated with mammary inflammatory markers, and butyrate administration attenuated LGE-induced mastitis in mice. Mechanistically, butyrate promotes M2 macrophage polarization and inhibits NF-κB and NLRP3 inflammasome activation via G-protein-coupled receptor 41 (GPR41). Collectively, our results demonstrate that gut microbiota from PS-dosed mice alleviate LGE-induced mastitis in mice by promoting SCFA production and regulating macrophage polarization. Our findings provide deeper insights into gut dysbiosis-associated mastitis and highlight the potential of modulating gut microbiota and its metabolism as a strategy for managing mastitis and other related diseases.