Renal-clearable drug delivery systems (DDSs) offer significant advantages compared to conventional non-renal-clearable DDSs due to their reduced toxicity and enhanced therapeutic efficacy. However, despite the development of renal-clearable DDSs in the past decade, deeper understanding of how the biological barriers, especially the intracellular barriers affect their therapeutic efficiency remain poorly explored. Herein, the antitumor efficiency and the intracellular behavior of renal-clearable Au-DOX which use renal-clearable gold nanoparticles (AuNPs) as delivery vectors for doxorubicin (DOX) were systematically investigated. The results revealed that although the toxicity of Au-DOX was significantly lower than that of free DOX due to efficient elimination of off-target DOX through renal clearance, the altered cellular uptake pathway compromised the antitumor efficacy of Au-DOX. Most Au-DOX was endocytosed and sequestered within lysosomes, preventing it from diffusing into nucleus to elicit therapeutic effect. Our results indicate that the lysosomal barrier induced ineffective intracellular delivery would counteract the therapeutic efficacy of renal-clearable DDSs and highlight the role of overcoming intracellular barriers when designing DDSs.

Obesity development is linked to disturbances in the gut microbiota. Inonotus obliquus polysaccharides (IOPs) have potential therapeutic efficacy in alleviating metabolic disorders. However, the mechanism by which IOP prevents obesity via regulating gut microbiota remains elusive. IOP was extracted and structurally characterized by FT-IR and NMR spectroscopy, confirming typical polysaccharide structures. Structurally, IOP is a 5.4 kDa polysaccharide predominantly composed of glucose, galactose, xylose, mannose, galacturonic acid, glucuronic acid, as well as rhamnose, arabinose, and methyl-galactose. Administration of IOP to high-fat diet (HFD)-fed mice effectively curtailed weight gain and improved serum lipid parameters. Furthermore, it mitigated lipid deposition within hepatic and adipose tissues, while successfully countering HFD-triggered liver damage. Notably, IOP induced significant changes in microbial diversity and composition by selectively increasing the abundance of Streptococcaceae while suppressing Faecalibaculum rodentium at the family and species levels. These findings highlight that IOP is a promising functional food ingredient that regulates gut microbiota for obesity prevention.

Triple-negative breast cancer (TNBC) is an aggressive breast cancer (BC) subtype with limited benefit from immune checkpoint blockade (ICB) and frequent immune-related adverse events (irAEs). We achieved stable overexpression of PD-1 in tumor cells through intratumoral injection of a PD-1-overexpressing plasmid. The results indicated that intratumoral PD-1 gene therapy exerted a dual effect. It significantly inhibited tumor cell growth via PD-1/PD-L1 interaction. Moreover, it promoted antitumor immunity, as evidenced by increased CD8+ T cells in the spleen, mesenteric lymph nodes (MLNs), and tumor tissues. Nevertheless, intratumoral PD-1 gene therapy induced intestinal side effects. Importantly, the probiotic Lactobacillus rhamnosus (L. rhamnosus) not only enhanced the antitumor activity of intratumoral PD-1 gene therapy but also alleviated intestinal injury. L. rhamnosus increased the expression of tight junction proteins (TJPs), including Claudin-1, Occludin, and ZO-1, upregulated anti-inflammatory cytokines (IL-10, TGF-β) and downregulated pro-inflammatory cytokines (IL-6, TNF-α). Notably, L. rhamnosus treatment also favorably reshaped the gut microbial composition, increasing the abundance of beneficial bacteria such as Lactobacillus, Bacteroides, and Parabacteroides, while reducing the levels of potential pathogenic bacteria including Prevotella, Staphylococcus, Adlercreutzia, and Desulfovibrio. Furthermore, metabolomic analysis revealed that L.rhamnosus significantly influenced tyrosine metabolism-related metabolites, downregulating the abundance of L-tyrosine, 3,4-dihydroxyphenylalanine (DOPA), N-acetyltyramine, methionyl-tyrosine, glutamic acid, and valine, while upregulating the level of butanoic acid. Collectively, these findings provide novel therapeutic strategies for enhancing the efficacy of TNBC treatment and alleviating intestinal side effects associated with immunotherapy.

Lactobacillus rhamnosus (L. rhamnosus) can modulate intestinal microbiota, decrease harmful bacterial metabolites, and thereby improve the intestinal microenvironment of patients with ulcerative colitis (UC). However, rapid inactivation and low colonization efficiency caused by intestinal peristalsis and impaired mucosa severely restrict its therapeutic outcomes. Extracellular vesicles (EVs) exhibit excellent mucus-penetrating ability that enables them to reach deep intestinal crypts. Subsequently, EVs directly deliver repair signals to intestinal epithelial cells and immune cells, effectively promoting intestinal mucosal repair. In this study, we prepared stemness-associated mouse embryonic fibroblast-derived extruded nanovesicles (sMEF-eNVs) via small-molecule intervention and three-dimensional (3D) culture. The prepared sMEF-eNVs displayed nanoscale morphology, EV-associated phenotype expression, and physicochemical features consistent with those of EVs. In a DSS-induced mouse model of UC, sMEF-eNVs improved epithelial barrier integrity, increased tight-junction and mucus-associated barrier signals, and attenuated mucosal inflammatory responses. Combined administration of sMEF-eNVs and L. rhamnosus further alleviated disease activity, improved histological injury, modulated Th17/Treg-associated immune imbalance, and was accompanied by shifts in gut microbial composition and fecal metabolic profiles. These findings support a vesicle-probiotic combination strategy for intestinal inflammation.

TNFRSF4 (OX40) -TNFSF4 (OX40L) axis is the core costimulatory pathway in the TNF/TNFR superfamily that regulates T cell responses. The binding of OX40L to OX40 on the surface of activated T cells significantly enhanced the proliferation and survival of CD4+ and CD8+ T cells and the secretion of IFN-γ and IL-2, while inhibiting the immunosuppressive activity of regulatory T cells, thereby amplifying the anti-tumor immunity. However, although OX40 agonist monotherapy is well tolerated in phase I/II clinical trials, the objective response rate is lower than that of PD-1 monotherapy, and it does not significantly prolong progression-free survival. At the mechanistic level, insufficient affinity, limited infiltration of T cells in the tumor and residual regulatory T cells are considered to be the main bottlenecks. Despite higher objective response rates with the addition of PD-1, radiotherapy, or chemotherapy, grade 3–4 immune-related adverse events were associated with higher rates. In the future, novel OX40 agonists with high affinity and selective activation in the tumor microenvironment should be developed or incorporated into the framework of combined immunotherapy as an adjuvant strategy to achieve a balance between efficacy and safety.

Post-translational modification (PTM) encompasses diverse modifications, including phosphorylation, methylation, ubiquitin-like modifications (UBLs), and so on, which profoundly influence cellular functions. UFMylation is a recently identified ubiquitin-like modification, which is mediated by the Ubiquitin-like Ubiquitin Fold Modifier 1 (UFM1) conjugation system. The UFM1 conjugation system comprises UFM1, Ubiquitin-like protein activating enzyme 5 (UBA5), UFM1-conjugating enzyme 1 (UFC1), UFM1-specific ligase 1 (UFL1), UFM1-specific protease 1 (UFSP1), UFM1-specific protease 2 (UFSP2), UFM1-binding protein 1 (UFBP1), and CDK5 regulatory subunit-associated protein 3 (CDK5RAP3). Accumulating research has demonstrated that the UFM1 conjugation system regulates various cellular stress responses, including endoplasmic reticulum (ER) stress, protein trafficking, DNA damage repair, and autophagy. Additionally, abnormal stress adaptations of the UFM1 conjugation system contribute to the pathophysiological complications of inflammatory diseases and cancer, underscoring its significance as a key regulatory node in human health and disease. Therefore, this review provides a comprehensive exploration of the structural characteristics of UFM1 conjugation system members and the mechanistic roles of UFMylation by UFM1 conjugation system-mediated diseases related to cellular stress responses, which will not only facilitate the identification of novel diagnostic and prognostic indicators but also enable the identification of specific therapeutic targets for UFM1 conjugation system-related diseases.

1,1-Dimethylbiguanide hydrochloride (Metformin) effectively lowers blood glucose levels in type 2 diabetes mellitus (T2DM) patients, but often causes gastrointestinal side effects, possibly due to disrupted bile acid metabolism. Bacillus subtilis, a key fermentative strain in traditional Chinese fermented foods like douchi, has been shown to aid in blood glucose control in T2DM mice, although the mechanism remains unclear. We induced T2DM in male C57BL/6J mice using a 60 % high-fat diet (HFD) and 75 mg/kg streptozotocin (STZ). Mice were administered PBS, metformin (200 mg/kg/day), Bacillus subtilis (10(10) CFU/mL), or their combination (Metformin + B. subtilis) via oral gavage for six weeks. The results indicated that B. subtilis significantly increased the quantities and compositions of key bacteria associated with bile acid metabolism, including Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria compared to the metformin group (P < 0.05). B. subtilis also altered bile acid composition, promoting the conversion of primary bile acids like cholic acid (CA), muricholic acid (MCA), taurochenodeoxycholic acid (TCDCA), and glycochenodeoxycholic acid (GCDCA) into secondary bile acids, including lithocholic acid (LCA) and deoxycholic acid (DCA) (P < 0.05), thereby enhancing bile acid metabolism. Furthermore, B. subtilis significantly reduced bile acid accumulation and colon tissue damage induced by metformin, while enhancing its blood glucose-lowering effect and effectively mitigating gastrointestinal side effects (P < 0.05). These findings propose a novel therapeutic approach for T2DM.

Background: As the most disastrous tumor microenvironment of pancreatic cancer, nutrient deprivation determined various cancer cell biology, especially the cell death resistance. Our objective is to elucidate the role of nutrient deprivation in ferroptosis resistance of pancreatic cancer cells and to explore potential therapeutic strategies to overcome it. Methods: To replicate the nutrient-deprived tumor microenvironment, pancreatic cancer cell lines (PANC1 and Patu8988T) were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 2% fetal bovine serum (FBS). Ferroptosis was assessed by Cell Counting Kit-8 (CCK8), Malondialdehyde (MDA) assay, and C11 BODIPY staining. The signaling activity was assessed via western blot and quantitative reverse transcription polymerase chain reaction (qRT-PCR), respectively. Results: Ferroptosis inducers promoted pancreatic cancer cell death could be significantly reversed under nutrient deprivation condition. Nutrient deprivation upregulated the expression of SREBP1 and SCD1, leading to increased intracellular levels of monounsaturated fatty acids (MUFAs). Genetic knockdown of SREBP1 or SCD1, as well as treatment with rapamycin (an mTOR inhibitor), reversed the nutrient deprivation induced increase in mature SREBP1 and SCD1 expression and restored lipid peroxidation both in vitro and in vivo. The synergistic application of sorafenib and rapamycin yielded a profoundly efficacious therapeutic outcome in vivo. Conclusion: Our findings demonstrate that nutrient-deprived pancreatic cancer cells adaptively enhance MUFA biosynthesis through the SREBP1-SCD1 axis, conferring resistance to ferroptosis. This resistance can be effectively overcome by combination therapy with sorafenib and rapamycin, offering a promising strategy to target the ferroptosis vulnerability shaped by the pancreatic tumor microenvironment.

Following the publication of the above article, the authors drew to the Editor's attention that the image in Fig. 3A on p. 1356 for the 'Migration/BxPC3/sh‑EGFP' experiment was mistakenly presented. This error arose as a consequence of a mistake that was made during the preparation of the final images. Furthermore, upon performing an independent analysis of the data in this paper in the Editorial Office, it came to light that, for the colony‑formation assay experiments shown in Fig. 2F on p. 1355, the image selected for the 'PaTu8988/Flag‑Furin' experiment had already appeared in a different context in another paper published by the same authors, also in the journal International Journal of Oncology. After having examined their original data, the authors realize that this second figure in the paper had also been inadvertently assembled incorrectly. The revised versions of Fig. 2 (now showing the data correctly for the for the 'PaTu8988/Flag‑Furin' experiment) and Fig. 3 (showing the correct data for the 'Migration/BxPC3/sh‑EGFP' experiment) are shown on the next two pages. Note that the errors made during the compilation of these figures did not affect the overall results and conclusions reported in the paper. The authors are grateful to the Editor of International Journal of Oncology for granting them the opportunity to publish this corrigendum, and all the authors agree with its publication; furthermore, they apologize to the readership of the journal for any inconvenience caused. [International Journal of Oncology 50: 1352‑1362, 2017; DOI: 10.3892/ijo.2017.3896].

To investigate the contribution of individual arginines, we employed site-directed mutagenesis to generate arginine-to-alanine (R→A) substitution mutations in the N-terminal domain of Forkhead box M1 (FoxM1). The R15A mutation impaired FoxM1 transcriptional activity, hindered FoxM1 nuclear translocation and failed to promote the migratory and invasive behavior of glioma cells than other single arginine mutations. Furthermore, we demonstrated that FoxM1 expression was associated with Mitochondrial transcription factor A (TFAM) expression. Overexpressing FoxM1 increased TFAM protein levels, which was reversed by FoxM1 knockdown in glioblastoma multiforme (GBM) cells. The siRNA-mediated reduction of TFAM expression was rescued by FoxM1 overexpression. Also, FoxM1 overexpression promoted TFAM promoter luciferase activity. Importantly, the R15A mutation failed to promote TFAM expression. Additionally, FoxM1 increased the expression of mitochondrial fusion markers, Optic atrophy protein 1 (OPA1) and Mitofusin 1 (MFN1) and led to interconnected mitochondria, while FoxM1 knockdown reversed this effect. Moreover, FoxM1 promoted mitochondrial fission markers, Dynamin-related protein 1 (DRP1), Mitochondrial fission factor (MFF) and Mitochondrial fission protein 1 (FIS1). Notably, the R15A mutation resulted in loss of FoxM1 regulation of fusion and fission-related protein expression. Taken together, our findings reveal that that the N-terminal arginine 15 is a key site for the transcriptional activation and function of FoxM1 in GBM cells, suggesting its potential as a therapeutic target in GBM.

Background Under the strategic framework of "Healthy China 2030", medical education must transition from a "Treatment-centered" paradigm to a "Prevention-Disease-Rehabilitation" holistic health model. Traditional medical education faces six core challenges: outdated training objectives, fragmented curricula, insufficient practical innovation, weak interdisciplinary integration, inadequate humanistic cultivation, and delayed competency adaptation. To address these issues, Jiangsu University has leveraged its comprehensive university resources since 2016. Methods Jiangsu University developed the "One Core with Dual Engines, Quintessential Competency Cultivation" clinical talent training system, which synergizes four key components: curricular integration to break disciplinary silos, pedagogical revolution to modernize teaching approaches, learning innovation to enhance practical skills, and humanistic integration to strengthen patient-centered values. Results The implementation of this model has successfully achieved three key outcomes: (1) implementation of comprehensive curricular restructuring on the "cell-tissue-organ-system" continuum, which transforms16 discrete basic medical courses (69.5 credits/1048 hours) into 15 integrated organ system-based courses (51 credits/976 hours); (2) highergraduate program matriculation rates (49.09% overall; 76.06% in clinical medicine) and National Medical Licensing Examination pass rates (94.88% in 2023, surpassing national averages); and (3) markedly improved satisfaction, as evidenced by 98% employer satisfaction and 94% "excellent" ratings for graduate competency in national surveys. Conclusion By aligning medical education with societal health demands, the "Jiangsu University Paradigm" provides a replicable model for cultivating holistic healthcare professionals, offering valuable insights for medical education reform in China and beyond.

Glioblastoma multiforme (GBM), whose pathogenesis involves proneural-to-mesenchymal transition (PMT), is the most malignant type of glioma and is associated with a bleak prognosis. Lactate dehydrogenase (LDH) comprises two major subunits, LDHA and LDHB, which can assemble into five different isoenzymes (LDH1-5). However, the role of LDH isoenzyme and its subunits in different GBM subtypes is largely unknown. Our findings reveal that LDHA and LDHB subunits correlated with mesenchymal and proneural subtype classification, and have prognostic and clinical significance in GBM patients. Moreover, it is demonstrated that LDH5, characterized by high LDHA and low LDHB levels, is highly expressed in mesenchymal subtype GBM cells and promotes proliferation, migration, and PMT. Conversely, proneural subtype GBM cells exhibited LDH1 dominance, and low LDHA and high LDHB levels. Notably, LDH1 played a pivotal role in the proliferation, migration, and PMT of proneural glioma cells. For treatment of proneural subtype GBM, gossypol-acetic acid (GAA)-bovine serum albumin (BSA) nanoparticles (GAA-BSA NPs) were developed to ameliorate PMT by targeting LDH1. These nanoparticles effectively suppress proneural subtype tumor growth both in vitro and in vivo, surpassing their efficacy against the mesenchymal subtype. The results offer several novel insights into the role of LDH isoenzyme in subtype classification between mesenchymal and proneural GBM and provide a promising therapeutic approach for proneural subtype GBM.

Tumor vaccine, which can effectively prevent tumor recurrence and metastasis, is a promising tool in tumor immunotherapy. However, heterogeneity of tumors and the inability to achieve a cascade effect limit the therapeutic effects of most developing tumor vaccine. We have developed a cascading immunoinducible in-situ mannose-functionalized polydopamine loaded with imiquimod phenylboronic hyaluronic acid nanocomposite gel vaccine (M/P-PDA@IQ PHA) through a boronic ester-based reaction. This reaction utilizes mannose-functionalized polydopamine loaded with imiquimod (M/P-PDA@IQ NAs) as a cross-linking agent to react with phenylboronic-grafted hyaluronic acid. Under near-infrared light irradiation, the M/P-PDA@IQ PHA caused local hyperthermia to trigger immunogenic cell death of tumor cells and tumor-associated antigens (TAAs) releasing. Subsequently, the M/P-PDA@IQ NAs which were gradually released by the pH/ROS/GSH-triggered degradation of M/P-PDA@IQ PHA, could capture and deliver these TAAs to lymph nodes. Finally, the M/P-PDA@IQ NAs facilitated maturation and cross-presentation of dendritic cells, as well as activation of cytotoxic T lymphocytes. Overall, the M/P-PDA@IQ PHA could serve as a novel in situ vaccine to stimulate several key nodes including TAAs release and capture, targeting lymph nodes and enhanced dendritic cells uptake and maturation as well as T cells activation. This cascading immune activation strategy can effectively elicit antitumor immune response.

Although methylguanine-DNA-methyltransferase (MGMT) plays an important role in resistance to temozolomide (TMZ) in glioma, 40% of gliomas with MGMT inactivation are still resistant to TMZ. The underlying mechanism is not clear. Here, we report that forkhead box M1 (FoxM1) transcriptionally activates the expression of DNA repair gene replication factor C5 (RFC5) to promote TMZ resistance in glioma cells independent of MGMT activation. We showed that RFC5 expression is positively correlated with FoxM1 expression in human glioma cells and FoxM1 is able to transcriptionally activate RFC expression by interaction with the RFC5 promoter. Furthermore, knockdown of FoxM1 or RFC5 partially re-sensitizes glioma cells to TMZ. Consistently, thiostrepton, a FoxM1 inhibitor, in combination with TMZ significantly inhibits proliferation and promotes apoptosis in glioma cells. Taken together, these findings suggest that the FoxM1-RFC5 axis may mediate TMZ resistance and thiostrepton may serve as a potential therapeutic agent against TMZ resistance in glioma cells.

Polycystic ovary syndrome (PCOS) involves complex genetic, metabolic, endocrine, and environmental factors. This study explores the effects of nicotinamide mononucleotide (NMN) in a letrozole-induced PCOS mouse model, focusing on metabolic regulation. Letrozole-induced aromatase inhibition elevated androgen and reduced bile acid levels, linking liver dysfunction and gut imbalance to PCOS. Letrozole-treated mice exhibited disrupted estrous cycles, ovarian congestion, and elevated testosterone. NMN intervention alleviated hyperandrogenism, ovarian abnormalities, and bile acid decline but did not fully restore the estrous cycle or improve lipid profiles. Metabolomic analysis showed that NMN partially reversed bile acid and lipid metabolism disturbances. These findings highlight NMN’s protective role in reducing hyperandrogenism and ovarian cyst formation. However, effective PCOS treatment should target liver and gut metabolism, not just ovarian symptoms, to mitigate systemic effects. Bile acid dysregulation may play a key role in PCOS progression and warrants further investigation.

Background: In spite of numerous existing bio-surveillance systems for predicting glioma (GBM) prognosis, enhancing the efficacy of immunotherapy remains an ongoing conundrum. The continual scrutiny of the dynamic interplay between the sphingolipid metabolic pathway and tumor immunophenotypes has unveiled potential implications. However, the intricate orchestration of functional and regulatory mechanisms by long non-coding RNAs (lncRNAs) in GBM, particularly in the context of sphingolipid metabolism, remains cryptic. Methods: We harnessed established R packages to intersect gene expression profiles of GBM patients within the The Cancer Genome Atlas (TCGA) database with the compilation of sphingolipid metabolism genes from GeneCards. This enabled us to discern markedly distinct lncRNAs, which were subsequently deployed to construct a robust prognostic model utilizing Lasso-Cox regression analysis. We then scrutinized the immune microenvironment across various risk strata using the ssGSEA and CIBERSORT algorithms. To evaluate mutation patterns and drug resistance profiles within patient subgroups, we devised the "Prophytic" and "Maftools" packages, respectively. Results: Our investigation scrutinized lncRNAs linked to sphingolipid metabolism, utilizing glioma specimens from TCGA. We meticulously curated 1224 sphingolipid-associated genes gleaned from GeneCards and pinpointed 272 differentially expressed mRNAs via transcriptomic analysis. Enrichment analyses underscored their significance in sphingolipid processes. A prognostic model founded on 17 meticulously selected lncRNAs was systematically constructed and validated. This model adeptly stratified GBM patients into high- and low-risk categories, yielding highly precise prognostic insights. We also discerned correlations between immune cell infiltration and genetic mutation discrepancies, along with distinct therapeutic responses through drug sensitivity analysis. Notably, computational findings were corroborated through experimental validation by RT-PCR. Conclusion: In summation, our exhaustive inquiry underscores the multifaceted utility of the sphingolipid metabolic pathway as an autonomous diagnostic and prognostic indicator for glioma patients. Furthermore, we amalgamate a profusion of substantiated evidence concerning immune infiltration and gene mutations, thereby reinforcing the proposition that sphingolipid metabolism may function as a pivotal determinant in the panorama of immunotherapeutic interventions.

Extracellular vesicles released by probiotics have been demonstrated to effectively alleviate intestinal inflammation, yet the precise underlying mechanisms remain unclear. In this research, for the first time, Lactobacillus plantarum UJS001 (LP-UJS) was isolated from fermented sauerkraut in Zhenjiang, China. Thereafter, the therapeutic effect of LP-UJS-derived extracellular vesicles (LP-UJS-EVs) on dextran sulfate sodium-induced ulcerative colitis (UC) in mice was analyzed to elucidate the immune mechanisms. According to our findings, LP-UJS-EVs played a pivotal role in restoring the intestinal barrier and alleviating intestinal inflammation. Notably, LP-UJS-EVs induced M2 polarization of macrophages, promoted the release of IL-10 and TGF-β, inhibited the release of histamine, IL-6, and TNF-α, and exerted regulatory effects on intestinal microflora, as evidenced by the reduced abundances of Coprococcus, Parabacteroides, Staphylococcus, and Allobaculum, alongside the enhanced abundance of Prevotella. Furthermore, both LP-UJS and LP-UJS-EVs affected the lysine degradation pathway and significantly increased the abundance of related metabolites, especially oxoadipic acid. In summary, our results underscore the substantial therapeutic potential of LP-UJS and its secreted EVs in the treatment of UC.

Natural Nicotinamide Adenine Dinucleotide (NAD(+)) precursors have attracted much attention due to their positive effects in promoting ovarian health. However, their target tissue, synthesis efficiency, advantages, and disadvantages are still unclear. This review summarizes the distribution of NAD(+) at the tissue, cellular and subcellular levels, discusses its biosynthetic pathways and the latest findings in ovary, include: (1) NAD(+) plays distinct roles both intracellularly and extracellularly, adapting its distribution in response to requirements. (2) Different precursors differs in target tissues, synthetic efficiency, biological utilization, and adverse effects. Importantly: tryptophan is primarily utilized in the liver and kidneys, posing metabolic risks in excess; nicotinamide (NAM) is indispensable for maintaining NAD(+) levels; nicotinic acid (NA) constructs a crucial bridge between intestinal microbiota and the host with diverse functions; nicotinamide riboside (NR) and nicotinamide mononucleotide (NMN) increase NAD(+) systemically and can be influenced by delivery route, tissue specificity, and transport efficiency. (3) The biosynthetic pathways of NAD(+) are intricately intertwined. They provide multiple sources and techniques for NAD+ synthesis, thereby reducing the dependence on a single molecule to maintain cellular NAD(+) levels. However, an excess of a specific precursor potentially influencing other pathways. In addition, Protein expression analysis suggest that ovarian tissues may preferentially utilize NAM and NMN. These findings summarize the specific roles and potential of NAD(+) precursors in enhancing ovarian health. Future research should delve into the molecular mechanisms and intervention strategies of different precursors, aiming to achieve personalized prevention or treatment of ovarian diseases, and reveal their clinical application value.

Background Gastric cancer (GC) is a common malignancy and a leading cause of cancer-related death with high morbidity and mortality. Methyl-CpG binding domain protein 3 (MBD3), a key epigenetic regulator, is abnormally expressed in several cancers, participating in progression and metastasis. However, the role of MBD3 in GC remains unknown. Methods MBD3 expression was assessed via public databases and validated by western blotting and quantitative real-time polymerase chain reaction (qRT-PCR). The prognosis of MBD3 was analysed via bioinformatics based on the TCGA dataset. The migration, invasion and proliferation of GC cells were examined by transwell, wound healing, cell counting kit (CCK)-8, colony-formation and xenograft mouse models. Epithelial-mesenchymal transition (EMT) and phosphatidylinositide 3-kinases/ protein Kinase B (PI3K/AKT) pathway markers were evaluated by Western blotting. RNA sequencing was used to identify the target of MBD3. Results MBD3 expression was higher in GC tissues and cells than in normal tissues and cells. Additionally, high MBD3 levels were associated with poor prognosis in GC patients. Subsequently, we proved that MBD3 enhanced the migration, invasion and proliferation abilities of GC cells. Moreover, western blot results showed that MBD3 promoted EMT and activated the PI3K/AKT pathway. RNA sequencing analysis showed that MBD3 may increase actin γ1 (ACTG1) expression to promote migration and proliferation in GC cells. Conclusion MBD3 promoted migration, invasion, proliferation and EMT by upregulating ACTG1 via PI3K/AKT signaling activation in GC cells and may be a potential diagnostic and prognostic target.