Colorectal cancer (CRC) remains one of the most common and lethal malignancies worldwide, with limited effective biomarkers for predicting prognosis and guiding immunotherapy response. The present study aimed to investigate the potential of DDB1- and CUL4-associated factor 13 (DCAF13) as a biomarker for CRC. Bioinformatics analyses were performed using public datasets from The Cancer Genome Atlas and Gene Expression Omnibus; ESTIMATE, CIBERSORT, immune checkpoint, TIDE score and Kaplan-Meier survival analyses were performed to assess the prognostic value and implications of DCAF13 expression in CRC. In silico findings were validated through immunohistochemistry (IHC), in vitro cell-based assays and transcriptomic analysis. Increased DCAF13 expression levels were associated with reduced overall survival in patients with CRC based on the bioinformatics analysis, and this was validated using samples from patients using IHC. Immune profiling demonstrated an increased infiltration of M0 and M1 macrophages, activation of mast cells, neutrophils and CD4+ memory T cells in the DCAF13-high expression group compared with the DCAF13-low expression group. DCAF13 expression was correlated with immune modulators and checkpoint genes. High DCAF13 expression was associated with lower TIDE scores. In vitro assays and transcriptomic analyses confirmed the pro-tumourigenic effects of DCAF13, which demonstrated roles in regulating cell proliferation, migration, clonogenicity, adhesion, metastasis, epithelial-to-mesenchymal transition and homologous recombination in CRC cells. The present study demonstrated that DCAF13 was upregulated in CRC and served a role in tumour progression, thus providing novel insights into the pro-tumourigenic functions of DCAF13 and its potential as a critical regulator in CRC.

AbstractBackground: Cancer-associated fibroblasts (CAFs) critically regulate colorectal cancer (CRC) progression, but the contribution of 5-fluorouracil (5-FU)-induced senescent CAFs to CRC malignancy and whether this stromal response can be pharmacologically restrained remain unclear. This study investigated whether Glabridin (Gla) attenuates the pro-tumorigenic activity of 5-FU-induced senescent CAFs.Methods: Primary human CRC-derived CAFs were exposed to 5-FU to establish a senescence model, which was validated by SA-β-Gal staining, EdU assay, cell-cycle analysis, γ-H2AX immunofluorescence, p53/p21 detection, and SASP marker analysis. Drug-free conditioned media (CM) collected from control, 5-FU-treated, and 5-FU plus Gla-treated CAFs after drug withdrawal and medium replacement were applied to RKO and SW480 cells to assess proliferation, migration, and invasion. Transcriptomic and integrative analyses were used to identify candidate SASP factors, followed by RT-qPCR and ELISA validation.Results: 5-FU induced a stable senescent phenotype in primary CAFs and enhanced SASP factor expression. CM from senescent CAFs promoted CRC cell proliferation, wound closure, and invasion. MMP1, MMP3, and MMP12 were identified as candidate senescence-associated secretory factors and were upregulated in 5-FU-induced senescent CAFs. Gla significantly suppressed 5-FU-induced MMP1/3/12 mRNA expression and reduced the cell number-normalized secretion of these MMPs in senescent CAF-CM. Functionally, CM from 5-FU plus Gla-treated CAFs exhibited weaker pro-tumorigenic effects than 5-FU-CM.Conclusions: Gla attenuates the pro-tumorigenic activity of 5-FU-induced senescent CAFs, at least in part by suppressing MMP1/3/12 expression and secretion. These findings suggest that Gla may help mitigate chemotherapy-associated stromal remodeling in CRC.

Gurigumu-13 (GR) was a classical Mongolian medicinal compound widely used in clinical practice for the treatment of non-alcoholic fatty liver disease (NAFLD). This study aimed to investigate the underlying mechanism by which GR exerted its therapeutic effects against NAFLD. The main active components of GR were identified using high-resolution mass spectrometry. Using network pharmacology approaches, key targets and potential pathways of GR against NAFLD were predicted and preliminarily validated through molecular docking. Furthermore, transcriptomic sequencing analysis, combined with in vivo and in vitro experiments was performed to elucidate the efficacy and mechanism of GR in alleviating NAFLD. In vivo, a rat model of NAFLD was established by feeding Wistar rats a methionine-choline deficient (MCD) diet. Serum levels of AST, ALT, TC, TG, MDA, GSH, SOD, HDL-C, and LDL-C were measured. Liver tissues were collected for histopathological examination via H E staining. In vitro, HepG2 cells were treated with oleic acid and palmitic acid to induce lipid accumulation. Intracellular TG content, as well as MDA and SOD levels, were assessed. Lipid deposition was visualized by Oil Red O staining. The expression of relevant genes and proteins was further analyzed by Western blotting and quantitative real-time PCR (qRT-PCR). Mass spectrometry analysis identified a total of 151 major bioactive compounds in GR. Through network pharmacology, 406 effective active ingredients and 1,012 potential targets of GR were screened. After intersecting with NAFLD-related genes (1,929 genes collected from Genecards and OMIM databases), 329 common targets were obtained. Protein-protein interaction network analysis indicated that STAT3, AKT, and others served as core targets. GO and KEGG enrichment analyses revealed that these genes are primarily involved in biological processes such as oxidative stress and lipid metabolism, as well as signaling pathways including PI3K/AKT and insulin resistance. In vivo, after 8 weeks of MCD diet feeding, rats in the model group exhibited significant hepatocellular fat accumulation, along with increases in body weight (1.82 times of the control group) and liver index (p < 0.05). GR intervention ameliorated these pathological changes, reduced serum levels of AST, ALT, TC, TG, MDA, and LDL-C (p < 0.05), and elevated levels of GSH, SOD, and HDL-C (p < 0.05). In vitro, GR significantly decreased triglyceride content in lipid-loaded HepG2 cells, alleviated intracellular lipid deposition (p < 0.05), and reduced the elevated glucose concentration in the culture medium of the model group. Additionally, GR reversed the increase in ROS and MDA contents and the decrease in SOD activity induced by the lipid-accumulation model. Western blotting results demonstrated that GR upregulated the phosphorylation levels of PI3K, AKT, and mTOR, and reduced STAT3 protein expression (p < 0.05). These changes were further supported by qPCR results (p < 0.05). The improvement of NAFLD by GR may be primarily achieved through its active components (such as ellagic acid and terchebin) via modulation of key targets (such as STAT3) and signaling pathways (such as PI3K/AKT/mTOR), thereby regulating lipid metabolism, alleviating oxidative stress and insulin resistance, and exerting therapeutic effects.

Ethnopharmacological relevanceBatri-7 (BT-7) is a classic Mongolian herbal formula traditionally valued for its anti-inflammatory properties. It has been utilized within Mongolian medicine for gastrointestinal disorders and inflammatory conditions, suggesting its potential to intercept colorectal carcinogenesis. However, its efficacy against colorectal adenoma (CA)—the primary precursor lesion of sporadic colorectal cancer (CRC)—and the underlying molecular mechanisms remain poorly understood.Aim of the studyThis study aims to systematically evaluate the efficacy and mechanism of BT-7 in suppressing CA progression by integrating network pharmacology, in vivo models, and patient-derived organoids.MethodsThe chemical profile of BT-7 was characterized by Liquid chromatography-mass spectrometry (LC-MS). Network pharmacology was employed to predict potential targets and pathways. An AOM/DSS-induced colitis-associated colorectal cancer (CAC) mouse model was used to assess in vivo efficacy. Patient-derived CA organoids were established and treated with BT-7 to evaluate its anti-adenoma effects in vitro. Transcriptomic analyses and functional rescue experiments were conducted to elucidate the underlying mechanisms.ResultsBT-7 treatment significantly suppressed adenoma formation and progression in AOM/DSS mice, alleviated colitis symptoms, and reduced proliferative activity. In patient-derived adenoma organoids, BT-7 inhibited growth and induced apoptosis in a dose- and time-dependent manner. Transcriptomic analyses consistently identified the PI3K/AKT pathway as a key target of BT-7. Functional studies confirmed that BT-7 downregulated PI3K/AKT signaling and specifically upregulated the pro-apoptotic protein BAX. Rescue experiments using PI3K and AKT activators (740Y-P and SC-79) reversed BT-7–induced growth inhibition and apoptosis, whereas an AKT inhibitor (MK-2206) phenocopied the pro-apoptotic effects of BT-7.ConclusionBT-7 effectively suppresses progression of CA by inhibiting the PI3K/AKT signaling pathway and activating BAX-mediated apoptosis. This study elucidates a defined molecular mechanism underlying BT-7’s action and highlights its potential as a multi-target natural agent for chemoprevention of CA.

Gastric cancer (GC) has a poor response to treatment, an unfavorable prognosis and a lack of reliable biomarkers for predicting disease progression and therapeutic outcomes. α-Ketoglutarate (α-KG) is a critical metabolite involved in cellular energy metabolism and epigenetic regulation during tumor development, which has emerged as a potential prognostic biomarker for GC. The present study aimed to explore this potential using publicly available datasets from The Cancer Genome Atlas and Gene Expression Omnibus databases to analyze α-KG-related genes and establish the α-KG Index (AKGI). By assessing the predictive performance of the AKGI model, the results demonstrated its capability to predict survival outcomes in patients with GC. Notably, high AKGI scores were associated with worse prognoses. Building on these findings, the associations between AKGI and clinical variables, immune cell infiltration and tumor mutation characteristics were assessed, further identifying potential therapeutic drugs for patients with high AKGI scores. Additionally, by analyzing signaling pathways and biological functions correlated with AKGI, the regulatory mechanisms and biological roles of α-KG in GC were elucidated. The findings of these analyses were further evaluated using cellular experiments, where α-KG treatment was demonstrated to significantly inhibit GC cell proliferation, migration and invasion. In conclusion, the present study successfully constructed and validated the AKGI as a potential prognostic biomarker for GC. The findings indicate that AKGI can identify patients likely to benefit from immunotherapy, enhance diagnostic precision and improve clinical outcomes in GC management. Moreover, AKGI offers a valuable framework for advancing the understanding of the role and mechanisms of α-KG in GC.

Matrigel is the most commonly used matrix for 3D organoid cultures. Research on the biomaterial basis of Matrigel for organoid cultures is a highly challenging field. Currently, many studies focus on Matrigel-based biological macromolecules or combinations to construct natural Matrigel and synthetic hydrogel scaffolds based on collagen, peptides, polysaccharides, microbial transglutaminase, DNA supramolecules, and polymers for organoid culture. In this review, we discuss the limitations of both natural and synthetic Matrigel, and describe alternative scaffolds that have been employed for organoid cultures. The patient-derived organoids were constructed in different cancer types and limitations of animal-derived organoids based on the hydrogel or Matrigel. The constructed techniques utilizing 3D bioprinting platforms, air-liquid interface (ALI) culture, microfluidic culture, and organ-on-a-chip platform are summarized. Given the potential of organoids for a wide range of therapeutic, tissue engineering and pharmaceutical applications, it is indeed imperative to develop defined and customized hydrogels in addition to Matrigel.

ETHNOPHARMACOLOGICAL RELEVANCE:Batri-7 (BT-7) is a compound herbal remedy rooted in traditional Mongolian medicine (TMM), composed of seven medicinal plants. Historically, it has been prescribed to treat various gastrointestinal conditions, including chronic enteritis and ulcerative colitis. However, despite this long-standing use in managing inflammatory disorders of the gut, the preventive effects of BT-7 on colitis-associated colorectal cancer (CAC), as well as its underlying molecular mechanisms, remains incompletely elucidated. AIM OF THE STUDY:This study aimed to assess the therapeutic efficacy of BT-7 in an AOM/DSS-induced murine model of CAC, while also investigating its modulatory effects on systemic inflammation, intestinal barrier function, gut microbiota dynamics, and inflammation-related molecular signaling pathways. MATERIALS AND METHODS:A chemically induced CAC model was established in C57BL/6 mice using azoxymethane (AOM) and dextran sodium sulfate (DSS). BT-7 was then administered orally at three dosage levels, and therapeutic outcomes were evaluated through a range of methods:clinical scoring, serum biochemical analysis, histological analysis, immunohistochemistry, and gut microbial profiling via 16S rRNA sequencing. To explore the underlying mechanistic pathways, with a particular focus on inflammasome regulation, transcriptome sequencing, quantitative PCR, and Western blotting were employed. RESULTS:BT-7 treatment was well-tolerated, with no evidence of liver or kidney toxicity. Mice receiving BT-7 exhibited improved body weight maintenance, lower disease activity scores, and preserved colon length. Histologically, there was substantial restoration of mucosal architecture and reduced epithelial hyperplasia, alongside significant suppression of tumor formation. Systemically, BT-7 reduced levels of key pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, and KC/GRO) and enhanced intestinal barrier integrity, as evidenced by the elevation of Occludin and ZO-1. Additionally, BT-7 decreased the abundance of pro-inflammatory taxa, such as Bacteroidota, and enriched beneficial genera like Lactobacillus and Dubosiella. The increased relative abundance of Lactobacillus_johnsonii and decreased Lactobacillus _murinus highlighted the probiotic effects of BT-7. Functional pathway prediction revealed that BT-7's microbiota was enriched in pathways related to carbohydrate and amino acid metabolism, as well as digestive system and immune-related functions. Transcriptomic analysis revealed downregulation of inflammatory signaling, particularly within the NOD-like receptor pathway, with reduced NLRP3 expression at both mRNA and protein levels confirming this effect. CONCLUSIONS:This work demonstrates that BT-7 exerts protective effects against CAC by suppressing inflammation, improving epithelial integrity, and restoring microbial homeostasis. Mechanistically, the NLRP3 inflammasome is identified as BT-7's primary target in this context, which aligns with its traditional use in treating gastrointestinal disorders. Collectively, these findings highlight BT-7's therapeutic potential as a chemopreventive agent, supporting its utility in aiding inflammation-associated colorectal cancer treatment.

Introduction:The theranostic potential of taurine-derived carbon dots (Tau/CDs) in colorectal cancer (CRC) remains largely unexplored, despite their promising physicochemical and biological properties. Methods:In this study, Tau/CDs were synthesized via a microwave-assisted irradiation method, employing citric acid as the carbon source, urea as the nitrogen source, and taurine (Tau) as the dopant. Comprehensive physicochemical characterization and biocompatibility assessments were performed both in vitro and in vivo. The anti-cancer efficacy of Tau/CDs against CRC was systematically evaluated through a series of functional assays, including cell viability, proliferation, migration, invasion, adhesion, clonogenicity, cell cycle progression, apoptosis, epithelial-mesenchymal transition (EMT), and transcriptomic profiling. The therapeutic efficacy was further validated in vivo using CRC xenograft murine models. Results:Tau/CDs exhibited excellent biocompatibility and significantly impaired key malignant properties of CRC cells, including viability, proliferation, migration, invasion, clonogenicity, and EMT. Treatment with Tau/CDs induced cell cycle arrest and apoptosis in vitro, while in vivo administration robustly suppressed tumor growth in xenograft models. Mechanistically, transcriptomic analysis combined with ferroptosis profiling identified Heme Oxygenase 1 (HO-1)-mediated ferroptosis as a critical pathway underlying the anti-tumor activity of Tau/CDs. Conclusion:Microwave-assisted synthesis of Tau/CDs from citric acid, urea, and Tau yielded biocompatible nanoparticles with potent anti-cancer properties. Tau/CDs were shown to inhibit CRC progression by regulating multiple malignant phenotypes, with HO-1-mediated ferroptosis emerging as a critical mechanistic axis. These findings highlight Tau/CDs as a promising candidate for future clinical translation in CRC nanomedicine.

Ovarian tissue vitrification is a key strategy for fertility preservation, but remains hindered by oxidative damage and compromised tissue viability. This study evaluates the cytoprotective effects of glutathione (GSH) supplementation during the vitrification of mouse ovarian tissue, including autologous transplantation experiments. Mice ovaries were cryopreserved with 0, 2, 4, or 8 mM GSH, followed by comprehensive histological, biochemical, and molecular assessments, as well as autologous transplantation to assess functional recovery. GSH, particularly at 4 mM, significantly preserved follicular architecture, enhanced antioxidant enzyme activity, attenuated mitochondrial dysfunction, DNA damage, and apoptosis, and suppressed pro-inflammatory and fibrotic signaling pathways. GSH also restored angiogenic markers and improved endocrine function in ovarian grafts after autologous transplantation, as evidenced by ameliorative levels of estradiol, progesterone, anti-Müllerian hormone (AMH), luteinizing hormone (LH), and follicle-stimulating hormone (FSH). These findings support GSH as a potent adjuvant for improving ovarian tissue vitrification outcomes and warrant further translational evaluation in fertility preservation protocols.

Colorectal cancer (CRC) is one of the most common malignant tumors worldwide. Although the use of small molecule drugs or targeted drugs has shown significant efficacy in the treatment of CRC, the drug resistance after treatment and the high recurrence and metastasis rate are the key obstacles affecting the success rate of treatment and survival of patients. Cellular senescence constitutes an important barrier to tumor progression. Senescent tumor cells and stromal cells are among the reasons for cancer treatment resistance. Different senescent programs can exert inhibitory or promotional effects on CRC. In serrated adenomas of colon, the senescence induced by intrinsic oncogenes serves as a threshold that precancerous lesions must traverse to develop into cancer. And the exposing of anti-cancer treatment, such as chemotherapy and radiotherapy, some cells also enter a senescent state, presenting a stable cell cycle arrest and senescence-associated secretory phenotype (SASP). SASP can activate immune surveillance but also contribute to the maintenance of cellular senescence microenvironment to help the CRC progression. Hence, in the pursuit of effective CRC treatment strategies, the issue of senescent cells is inevitable. By targeting features of senescent cells, such as upregulated anti-apoptotic signaling, altered metabolic signaling, and differential SASP secretion, depletion of senescent cells could be a promising strategy for the treatment of CRC. This review summarizes the endogenous and exogenous factors leading to cell senescence in CRC, as well as drug mechanisms, and focuses on the research progress of senescent tumors and stromal cells in CRC. Eventually, we discuss the strategies for CRC senescent cells after anti-cancer treatment to provide some theoretical basis and direction for retarding the malignant progression and recurrence of CRC.

Background The increasing prevalence of type 2 diabetes mellitus (T2DM) on a global scale has created a pressing demand for novel treatments. Gentianella turkestanorum (Gand.) Holub, a traditional Chinese herbal medicine, has been found to possess hypoglycemic effects. However, the mechanism of its action remains unclear. Objectives This study was to investigate the impact and mechanism of G. turkestanorum ’s water extract (WEG) in reducing insulin resistance (IR) in T2DM. Materials and Methods Db/db mice were administered WEG for 8 weeks, during which their body weight, blood glucose (BG), oral glucose tolerance test, islet tolerance test, fasting insulin, total cholesterol, triglyceride, high-density lipoprotein, and low-density lipoprotein were monitored. Additionally, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels in db/db mice were measured using enzyme-linked immunosorbent assay (ELISA). The study also evaluated the impact of WEG on liver injury through hematoxylin-eosin staining. The expression levels of key proteins and genes in both insulin signaling and inflammation-related pathways were detected using western blotting and real-time quantitative polymerase chain reaction. Results WEG has the potential to regulate glycolipid metabolism, reduce inflammation, and alleviate IR. The mechanism of action may involve promoting the insulin signaling pathway and inhibiting inflammation. Conclusion Gentianella turkestanorum could be a viable treatment option for T2DM and IR. Keywords , , chemical composition , T2DM , IR , inflammation

Identification of novel biomarkers for prediction of disease course and prognosis is needed to reduce morbidity of liver hepatocellular carcinoma (LIHC/HCC) patients. Although dysregulated Periodic tryptophan protein 1 homolog (PWP1/endonuclein) expression has been detected in several tumors, the potential regulatory effect of PWP1 on LIHC remains uncertain. Here we evaluated the expression of PWP1 using multiple online platforms, and demonstrated that PWP1 upregulation was consistently observed in LIHC relative to non-tumor liver tissues and correlated with unfavorable prognosis. Moreover, HCC prognosis was significantly influenced by the methylation status of various CpG sites in the PWP1 gene. Lastly, we provide direct evidence that PWP1 acts as a driver of HCC progression by showing that siRNA-mediated PWP1 silencing significantly suppressed HCC cell proliferation in vitro. These data strongly suggest that PWP1 silencing may be an effective therapeutic strategy to treat LIHC.

Introduction:The anti-cancer potency of copper-doped carbon quantum dots (Cu-CDs) against breast cancer progression needs more detailed investigations.Methods:With urea and ethylene glycol applied as carbon sources and copper sulfate used as a reactive dopant, Cu-CDs were synthesized in the current study by a one-step hydrothermal synthesis method, followed by the characterization and biocompatibility evaluations of Cu-CDs. Subsequently, the anti-cancer potency of Cu-CDs against breast cancer progression was confirmed by these biochemical, molecular, and transcriptomic assessments, including viability, proliferation, migration, invasion, adhesion, clonogenicity, cell cycle distribution, apoptosis, redox homeostasis, and transcriptomic assays of MDA-MB-231 cells.Results:The biocompatibility of Cu-CDs was confirmed based on the non-significant changes in the pathological and physiological parameters in the Cu-CDs treated mice, as well as the noncytotoxic effect of Cu-CDs on normal cells. Moreover, the Cu-CDs treatments not only decreased the viability, proliferation, migration, invasion, adhesion, and clonogenicity of MDA-MB-231 cells but also induced the redox imbalance, cell cycle arrest, and apoptosis of MDA-MB-231 cells via ameliorating the mitochondrial dysfunctions and regulating the MAPK signaling pathway.Conclusion:Our findings confirmed the biosafety and excellent anti-cancer potency of Cu-CDs against breast cancer progression by tapping into mechanisms that disrupt malignant behaviors and oxidative homeostasis of breast cancer cells.

Colorectal cancer (CRC) is a significant global health burden, with high morbidity and mortality rates. It is often diagnosed at middle to advanced stage, affecting approximately 35% of patients at the time of diagnosis. Currently, chemotherapy has been used to improve patient prognosis and increase overall survival. However, chemotherapy can also have cytotoxic effects and lead to adverse reactions, such as inhibiting bone marrow hematopoiesis, causing digestive dysfunction, hand-foot syndrome, and even life-threatening conditions. In response to these adverse effects, researchers have proposed using Traditional Chinese Medicine (TCM) as an option to treat cancer. TCM research focuses on prescriptions, herbs, and components, which form essential components of the current research in Chinese medicine. The study and implementation of TCM prescriptions and herbs demonstrate its distinctive holistic approach to therapy, characterized by applying multi-component and multi-target treatment. TMC components have advantages in developing new drugs as they consist of single ingredients, require smaller medication dosages, have a precise measure of pharmacodynamic effects, and have a clear mechanism of action compared to TCM prescriptions and herbs. However, further research is still needed to determine whether TMC components can fully substitute the therapeutic efficacy of TCM prescriptions. This paper presents a comprehensive analysis of the research advancements made in TCM prescriptions, herbs, and components. The findings of this study can serve as a theoretical basis for researchers who are interested in exploring the potential of TCM for the treatment of colorectal cancer.

Environmental osmolarity plays a crucial role in regulating the functions and behaviors of both host cells and pathogens. However, it remains unclear whether and how environmental osmotic stimuli modulate bacterial-host interfacial adhesion. Using single-cell force spectroscopy, we revealed that the interfacial adhesion force depended nonlinearly on the osmotic prestimulation of host cells but not bacteria. Quantitatively, the adhesion force increased dramatically from 25.98 nN under isotonic conditions to 112.45 or 93.10 nN after the host cells were treated with the hypotonic or hypertonic solution. There was a strong correlation between the adhesion force and the number of host cells harboring adherent/internalized bacteria. We further revealed that enhanced overexpression levels of collagen XV and II were responsible for the increases in interfacial adhesion under hypotonic and hypertonic conditions, respectively. This work provides new opportunities for developing host-directed antibacterial strategies related to interfacial adhesion from a mechanobiological perspective.

Regardless of the essential role of copper (Cu) in the physiological regulation process of mammalian reproduction, excessive exposure to Cu triggers the meiotic defects of porcine oocytes via compromising the mitochondrial functions. However, the connections between the excessive Cu exposure and meiotic defects of ovine oocytes have not been reported. In this study, the effect of copper sulfate (CuSO4) exposure on the meiotic potentials of ovine oocytes was analyzed. Subsequently, the ameliorative effect of glutathione (GSH) supplementation on the meiotic defects of CuSO4 exposed ovine oocytes was investigated. For these purposes, the in vitro maturation (IVM) of ovine cumulus oocyte complexes (COCs) was conducted in the presence of 5, 10, 20 and 40 μg/mL of CuSO4 supplementation. Subsequently, different concentrations of GSH (2, 4 and 8 mM) were added to the IVM medium containing CuSO4 solution. After IVM, the assay, including nuclear maturation, spindle organization, chromosome alignment, cytoskeleton assembly, cortical granule (CGs) dynamics, mitochondrial function, reactive oxygen species (ROS) generation, apoptosis, epigenetic modification and fertilization capacity of ovine oocytes were performed. The results showed that excessive Cu exposure triggered the meiotic defects of ovine oocytes via promoting the mitochondrial dysfunction related oxidative stress damage. Moreover, the GSH supplementation, not only ameliorated the decreased maturation potential and fertilization defect of CuSO4 exposed oocytes, but inhibited the mitochondrial dysfunction related oxidative stress damage, ROS generation, apoptosis and altered H3K27me3 expression in the CuSO4 exposed oocytes. Combined with the gene expression pattern, the finding in the present study provided fundamental bases for the ameliorative effect of GSH supplementation on the meiotic defects of CuSO4 exposed oocytes via inhibiting the mitochondrial dysfunctions, further benefiting these potential applications of GSH supplementation in the mammalian IVM system and livestock breeding suffering from the excessive Cu exposure.

The current study was designed to investigate the alleviative effect of lactoferrin interventions against the hepatotoxicity induced by titanium dioxide nanoparticles (TiO2-NPs). Thirty male Wistar rats were divided into six groups with 5 rats in each group. The first and second groups were intragastrically administered normal saline and TiO2-NPs (100 mg/kg body weight) as the negative control (NC) and TiO2-NP groups. The third, fourth, and fifth groups were intragastrically administered lactoferrin at concentrations of 100, 200, and 400 mg/kg body weight in addition to TiO2-NPs (100 mg/kg body weight). The sixth group was intragastrically administered Fuzheng Huayu (FZHY) capsules at a concentration of 4.6 g/kg body weight in addition to TiO2-NPs (100 mg/kg body weight) as the positive control group. After treatment for 4 weeks, the concentrations of lactoferrin were optimized based on the liver index and function results. Subsequently, the alleviative effects of lactoferrin interventions against TiO2-NP-induced hepatotoxicity in rat liver tissues, including the effects on histological damage, oxidative stress-related damage, inflammation, fibrosis, DNA damage, apoptosis, and gene expression, were investigated using histopathological, biochemical, and transcriptomic assays. The results showed that 200 mg/kg lactoferrin interventions for 4 weeks not only ameliorated the liver dysfunction and histopathological damage caused by TiO2-NP exposure but also inhibited the oxidative stress-related damage, inflammation, fibrosis, DNA damage, and apoptosis in the liver tissues of TiO2-NP-exposed rats. The transcriptomic results confirmed that the alleviative effect of lactoferrin interventions against the TiO2-NP exposure–induced hepatotoxicity was related to the activation of the PI3K/AKT signaling pathway.

IntroductionIn the present study, the synergistic protective effect of co-supplementation of glutathione (GSH) with selenium nanoparticles (SeNPs) on the cryopreservation efficiency of bull semen was analyzed.MethodsAfter collection, the ejaculates of Holstein bulls were subsequently diluted with a Tris extender buffer supplemented with different concentrations of SeNPs (0, 1, 2, and 4 μg/ml), followed by semen equilibration at 4°C and assessment of sperm viability and motility. Subsequently, the ejaculates of Holstein bulls were pooled, split into four equal groups, and diluted with a Tris extender buffer supplemented with basic extender (negative control group, NC group), 2 μg/ml SeNPs (SeNPs group), 4 mM GSH (GSH group), and 4 mM GSH plus 2 μg/ml SeNPs (GSH + SeNPs group). After cryopreservation, motility, viability, mitochondrial activity, plasma membrane integrity, acrosome integrity, concentration of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT), and ability of frozen-thawed sperm cells to support in vitro embryonic development were evaluated.Results and discussionNo side effect of SeNPs concentrations applied in the current study on the motility and viability of equilibrated bull spermatozoa was found. Meanwhile, supplementation of SeNPs significantly promoted the motility and viability of equilibrated bull spermatozoa. Furthermore, the co-supplementation of GSH with SeNPs effectively protected bull spermatozoa from cryoinjury as expressed by promoting semen motility, viability, mitochondrial activity, plasma membrane integrity, and acrosome integrity. Finally, the enhanced antioxidant capacity and embryonic development potential in the frozen-thawed bull spermatozoa cryopreserved by co-supplementation of GSH with SeNPs further confirmed the synergistic protective effect of co-supplementation of GSH with SeNPs on the cryopreservation of bull semen.

Objective:To investigate the effect and mechanism of lactoferrin (LF) on biological behaviors of brain glioma cells.Methods:The effect of LF at different concentrations (0, 100, 200 and 300 μg/mL) on the proliferative ability of human glioma cell line U87MG was analyzed by MTT assay to screen the optimal drug concentration. U87MG cells were divided into blank control group and LF (200 μg/mL) treatment group. Edu staining, Transwell assay, Mito-Tracker staining and DCFH-DA staining were used to detect the cell proliferation, migration, mitochondrial activity and reactive oxygen level. Autophagy marker protein microtubule associated protein 1 light chain 3B (LC3B) expression was detected by immunofluorescent chemical staining; contents of superoxide dismutase (SOD) and malonaldehyde (MDA) were detected by colorimetric chemical sensors; expressions of genes related to epithel-to-mesenchymal transformation (E-cadherin, fibronectin, N-cadherin, vimentin and Snail) were detected by reverse transcription (RT)-PCR; expressions of apoptosis-related proteins (Bax and Bcl-2) were detected by Western blotting.Results:Compared with blank control group, the LF treatment group had significantly decreased proportions of Edu positive cells and Mito-Tracker positive cells, number of cell migration and SOD content, and statistically increased proportion of DCFH-DA positive cells, MDA content and LC3B immunofluorescent staining intensity in cytoplasm ( P<0.05). Compared with blank control group, the LF treatment group had significantly decreased E-cadherin mRNA expression and Bcl-2 protein expression, and statistically increased fibronectin, N-cadherin, vimentin and Snail mRNA expressions and Bax protein expression ( P<0.05). Conclusion:LF can effectively inhibit the proliferation, migration, invasion and other biological behaviors of glioma cells, whose mechanism is closely related to mitochondrial activity.