Abstract Background Osteoarthritis (OA) frequently co-exists with type 2 diabetes mellitus (T2DM) owing to shared risk factors, including age and obesity, and disease severity is often greater in individuals with both conditions (OADM). However, molecular alterations associated with this co-morbidity remain incompletely understood. Here, we applied a proteomics-based approach to investigate glycated/ glycosylated synovial fluid (SF) proteins and to identify molecular patterns potentially linked to joint degeneration in OADM. Methods We performed the first global proteomic analysis of phenylboronic acid (PBA)-enriched cis-diol–containing glycation/ glycosylation-associated proteins from knee SF samples of OA and OADM patients. Enriched proteins were fractionated by SDS-PAGE and analyzed by LC-MS/MS, followed by quantitative SWATH-MS and bioinformatic pathway analyses. Selected proteins showing increased abundance in OADM were evaluated in SF and paired serum samples from a larger cohort using ELISA, and their discriminatory performance was explored using receiver operating characteristic (ROC) curve analysis. Results Three proteins, high-temperature requirement protein A1 (HTRA1), cathepsin G (CTSG), and alpha-1-acid glycoprotein 1 (AGP1), were consistently more abundant in OADM compared with OA. Functional annotation associated HTRA1 and CTSG with extracellular matrix remodeling and degradative processes, while AGP1 was linked to inflammatory responses. Exploratory ROC analyses in both SF and serum suggested that these proteins have potential utility in distinguishing OADM from OA within the studied cohort, with SF and serum levels showing significant concordance in OADM patients. Conclusions These findings suggest an association between T2DM-related metabolic dysregulation, synovial protein glycation/ glycosylation, and cartilage degeneration in OADM. HTRA1, CTSG, and AGP1 emerge as clinically measurable proteins with potential biomarker relevance for distinguishing OADM from OA, which may support improved disease stratification in future studies. Collectively, the results provide molecular insight into the co-pathogenesis of OA and T2DM and highlight glycation/ glycosylation-associated pathways as candidates for future precision-based therapeutic investigation, with the potential to mitigate OA severity in patients with T2DM and improve joint health.

INTRODUCTION:Primary hyperparathyroidism (PHPT) is a systemic endocrine disorder characterized by elevated PTH levels and hypercalcemia. Gene-specific epigenetic modifications like histone methylation play a pivotal role in PHPT by altering expression of parathyroid-specific genes. However, global histone modifications in parathyroid tumors and their implications for tumor behavior remain underexplored. EXPERIMENTAL DESIGN:We performed comparative histone modification profiling in blood and tissue samples from sporadic parathyroid adenomas (PA; n = 30), atypical parathyroid tumors (APT; n = 05), and parathyroid carcinomas (PC; n = 05) along with controls using histone H3 multiplex immunoassay. Significantly dysregulated modifications were validated by Western blotting (WB), and expression data was correlated with clinicopathological features. We performed pathway enrichment analysis to elucidate molecular pathways potentially linked to histone H3 modifications in parathyroid tumorigenesis. RESULTS:We observed dynamic alterations in histone H3 modifications of lysine (K) residues across parathyroid tumor phenotypes in blood and tissue samples. A reduction in H3K4 and H3K36 trimethylation (<60%) and an increase in H3K27 trimethylation and H3K9 monomethylation (>130%) compared to controls was observed. WB confirmed lower H3K4me3 (0.5 ± 0.4 vs 4.4 ± 0.7, P = .02; 0.4 ± 0.2 vs 4.4 ± 0.7, P = .004) and H3K36me3 (0.4 ± 0.2 vs 1.0 ± 0.3, P = .03; 0.2 ± 0.1 vs 1.0 ± 0.3, P = .02) in APT and PC than controls and higher H3K27me3 expression in PA (2.9 ± 1.2 vs 1.1 ± 0.21, P = .04), APT (4.2 ± 2.9 vs 1.1 ± 0.21, P = .007), and PC (6.9 ± 3.1 vs 1.1 ± 0.21, P = .0002) relative to controls. Pathway enrichment analysis identified molecular pathways, including calcium signaling, Wnt, and Hippo signaling, associated with histone H3 modifications. CONCLUSION:Increased expression of repressive H3K27me3 in blood and tissue samples, while decreased H3K4me3 and H3K36me3, suggest a shift toward transcriptional repression. Identifying their functional mechanisms may facilitate the discovery of novel insights for therapeutic targeting in parathyroid tumors.

Hepatocellular carcinoma (HCC), the predominant form of liver cancer, is the fifth most prevalent cancer worldwide and second leading cause of cancer-related deaths, underscoring its grim prognosis. Key risk factors for HCC include HBV, HCV, cirrhosis, inherited metabolic diseases, vitamin supplementation, heavy alcohol use, obesity etc. Despite the well-documented impact of alcohol on HCC, there remains a significant gap in understanding alcohol-associated HCC (A-HCC) compared to viral hepatitis associated HCC. So, in this study we tried to elucidate the role of UPR pathway in exacerbating HCC prognosis under the condition of A-HCC. Notably, our RT-qPCR and western blot analysis showed significant upregulation of PERK-ATF4-LAMP3 arm along with CHOP, VEGF-A and nuclear translocation of NF-KB in both HepG2 and Hep3B cell lines. Increased extracellular & intracellular cholesterol and triglyceride levels obtained can be related with the higher expression of SREBP-2 and SREBP-1c respectively. Ethanol exposure also enhanced the invasive and migratory properties of HCC, reduced apoptosis with increased stemness in a PERK dependent manner. Moreover, orally available PERK inhibitor (GSK2606414) successfully relieved the effects caused by ethanol in HepG2 and Hep3B cell lines. In summary, HCC cells gain aggressiveness due to ethanol exposure via PERK/ATF4/LAMP3 pathway, and targeting PERK could serve as a promising therapeutic strategy for A-HCC, mitigating several cancer hallmarks.

BACKGROUND:Folate (vitamin B9) is a water-soluble vitamin necessary for one-carbon metabolism, supporting the synthesis, repair, and methylation of DNA. While maternal folate status is well-studied for its role in fetal development and metabolic programming, the impact of inadequate folate intake in males on offspring development and metabolic diseases remains poorly understood. This study investigates the effects of folate deficiency in male parents on developing hepatic insulin resistance in offspring, focusing on molecular and metabolic disruptions within the liver. METHODS:Three-week-old C57BL/6 male mice were categorized into two groups: Group I received a folate-normal (FN) diet, and Group II was fed a folate-deficient (FD) diet for four weeks before mating. F1 offspring from Group I (FN diet) were mated to produce F2 offspring (PNMN: paternal normal, maternal normal). F1 males from Group II (lifetime FD diet) were mated with F1 females on an FN diet to produce F2 offspring (PDMN: paternal deficient, maternal normal). F2 offspring from both groups were maintained on an FN diet and monitored for body weight. The study assessed systemic markers of insulin resistance, lipid and glucose metabolism, and gene expression profiles and proteins associated with insulin signaling in the liver. Mechanistic pathways involving lipid-induced and ER stress-triggered hepatic insulin resistance were explored. RESULTS:Male offspring born to folate-deficient fathers (PDMN) exhibited significantly elevated fasting glucose and insulin levels, impaired glucose tolerance, increased HOMA-IR and reduced QUICKI at 10 weeks. Hepatic insulin signaling was disrupted, as evidenced by downregulated p-AKT levels in 7-week PDMN males. Lipogenic pathways were upregulated, with increased expression of transcription factors Srebf1c and Chrebp (both at gene and protein levels), contributing to hepatic steatosis. Gluconeogenic genes, including Foxo1 and Fbp1, were also upregulated, indicating elevated hepatic glucose output and exacerbation of hyperglycemia. Chronic endoplasmic reticulum (ER) stress, marked by upregulation of Perk and Atf6 (both at gene and protein levels), further impaired hepatic insulin signaling possibly by activating stress pathways and disrupting protein folding. CONCLUSION:This study provides the first evidence that paternal folate deficiency predisposes offspring to hepatic insulin resistance by disrupting insulin signaling, promoting lipid dysregulation, and activating ER stress pathways. These effects are more severe in males, underscoring sex-specific susceptibility. The findings emphasize the importance of balanced paternal folate intake during reproduction to prevent intergenerational metabolic disorders and suggest potential therapeutic targets to mitigate hepatic insulin resistance caused by paternal nutritional deficiencies.

Tissue biopsies are routinely used for identifying driver mutations in tumors but limitations such as tumor heterogeneity have encouraged the use of liquid biopsies for the identification of new biomarkers. ATP-citrate lyase (ACLY) has been recognized as a potential target for tumor inhibition due to its overexpression in several cancers. However, the clinical significance of this enzyme remains unknown in liquid biopsies. In this study, 31 pleural effusion fluid samples were collected from patients suspected of malignancy for analyzing the gene expression of ACLY and associated metabolic enzymes. Gene expression profiling by RT-PCR revealed significantly increased ACLY mRNA levels along with other metabolic enzymes (FASN, ACC, CS, and ICD1) in malignant lung adenocarcinoma pleural fluid samples indicating that ACLY plays a critical role in the progression of lung adenocarcinoma. This study identified the potential of using metabolic enzymes as biomarkers in identification of lung adenocarcinoma patients. The observations highlighted the significance of lipogenic enzymes mainly ACLY and fatty acid synthase as diagnostic markers in lung adenocarcinoma liquid biopsies.

Hepatocellular carcinoma (HCC), a leading cause of cancer-related mortality worldwide, is characterized by poor prognosis, high recurrence rates, and limited responsiveness to current therapies. Autophagy, a conserved catabolic pathway essential for cellular homeostasis, plays a paradoxical role in HCC, acting as a tumor suppressor during initiation but promoting survival and progression in advanced stages. Long non-coding RNAs (lncRNAs) have emerged as critical regulators of autophagy, influencing tumorigenesis, metastasis, and therapy resistance through mechanisms such as miRNA sponging, chromatin remodeling, and protein interactions. This review describes how autophagy contributes to HCC at different stages, outlines the dual functions of lncRNAs as oncogenic drivers or tumor suppressors, and illustrates their integration into key signaling networks of autophagy (e.g., PI3K/AKT/mTOR, AMPK, Beclin-1). LncRNAs have been shown to modulate drug resistance, including resistance to first-line agents, by altering autophagic flux and associated molecular pathways. We also explored emerging strategies for targeting the lncRNA–autophagy axis, such as siRNAs, antisense oligonucleotides, and CRISPR/Cas systems, that have shown promise in preclinical studies and may be adapted for HCC. Furthermore, autophagy-related lncRNAs hold potential as non-invasive diagnostic and prognostic biomarkers and as predictors of recurrence. Integrating multi-omics approaches to validate these candidates will be critical for translation into clinical practice. Collectively, this review highlights the lncRNA–autophagy network as a promising frontier of biomarker discovery for precision diagnostics and targets for innovative therapeutics. The regulatory role of lncRNAs in autophagy presents a paradigm shift, heralding new strategies for targeted treatment.

Osteoarthritis (OA) is the primary cause of joint impairment, particularly in the knee. The prevalence of OA has significantly increased, with knee OA being a major contributor whose pathogenesis remains unknown. Articular cartilage and the synovium play critical roles in OA, but extracting high-quality RNA from these tissues is challenging because of the high extracellular matrix content and low cellularity. This study aimed to identify the most suitable RNA isolation method for obtaining high-quality RNA from microquantities of guinea pig cartilage and synovial tissues, a relevant model for idiopathic OA. We compared the traditional TRIzol® method with modifications to spin column-based methods (TRIspin-TRIzol®/RNeasyTM, RNeasyTM kit, RNAqueousTM kit, and Quick-RNATM Miniprep Plus kit), and an optimized RNA isolation protocol was developed to increase RNA yield and purity. The procedure involved meticulous sample collection, specialized tissue processing, and measures to minimize RNA degradation. RNA quality was assessed via spectrophotometry and RT-qPCR. The results demonstrated that among the tested methods, the Quick-RNATM Miniprep Plus kit with proteinase K treatment yielded the highest RNA purity, with A260:280 ratios ranging from 1.9 to 2.0 and A260:230 ratios between 1.6 and 2.0, indicating minimal to no salt contamination and RNA concentrations up to 240 ng/μL from ⁓20 mg of tissue. The preparation, storage, homogenization process, and choice of RNA isolation method are all critical factors in obtaining high-purity RNA from guinea pig cartilage and synovial tissues. Our developed protocol significantly enhances RNA quality and purity from micro-quantities of tissue, making it particularly effective for RTqPCR in resource-limited settings. Further refinements can potentially increase RNA yield and purity, but this protocol facilitates accurate gene expression analyses, contributing to a better understanding of OA pathogenesis and the development of therapeutic strategies. Key features • Enables efficient RNA isolation from small, individual cartilage samples, eliminating the need for pooling and requiring minimal laboratory equipment. • Provides a reliable and cost-effective method for obtaining high-quality RNA suitable for RT-qPCR and gene expression analysis, from challenging tissue types like cartilage. Graphical overview Graphical representation comparing modified RNA isolation protocols for efficient RNA extraction from guinea pig cartilage and synovium.

The metabolic enzyme ATP citrate lyase is overexpressed in several cancers and links glucose metabolism with de novo fatty acid synthesis pathway by catalyzing the conversion of citrate into acetyl CoA and oxaloacetate. Potassium hydroxycitrate, its natural inhibitor, exhibits anticancer activity; however, its use is limited due to low bioavailability. This study aims to improve the efficacy of hydroxycitrate by its encapsulation in bovine milk exosome surface conjugated with folate for targeting lung cancer cells. The mean particle size of potassium hydroxycitrate-loaded exosomes (Exo-KH) and paclitaxel exosomes (Exo-Pac) was 183 nm and 174 nm; they had spherical morphology and encapsulation efficiency of 16.87 ± 2.78

The ketogenic diet (KD) is recognized for its broad therapeutic potential across metabolic, neurological, immunological, and aging-related domains; however, its efficacy and durability in females remain underexplored. This study investigated the effects of a short-term, intermittent KD on neurobehavioral function, lipid metabolism, immune response, and gut microbiota composition in sham-operated (intact) and ovariectomized (OVX, estrogen-deficient) female mice, aiming to delineate the interactions between diet and gonadal hormones. In sham females, KD reduced circulating estradiol levels and induced low-grade systemic inflammation, without producing measurable behavioral benefits. Conversely, KD exhibited reduced compulsivity, anxiety, and depression-like behaviors with improved well-being in OVX females, suggesting enhanced neurobehavioral resilience under estrogen-deficient conditions. Biochemical analyses revealed KD-induced hyperlipidemia in both groups, although the lipid load was attenuated in OVX mice. KD further decreased low-density lipoprotein cholesterol but led to uremia and histopathological evidence of hepatic steatosis and fibrosis in OVX females, despite unaltered liver function markers. 16S rRNA sequencing revealed hormone-dependent remodelling of the gut microbiota, characterised by distinct compositional shifts between the sham and OVX cohorts. Correlation analyses linked microbial alterations to improved lipid profiles and reduced anxiodepressive behaviors, implicating the gut-metabolic-brain axis in mediating the effects of KD. Collectively, these findings suggest that KD confers neurobehavioral and metabolic benefits in the absence of ovarian hormones but may also pose risks to the liver and kidneys. These results highlight ketogenic nutrition as a potential non-pharmacological strategy to mitigate menopause- and aging-associated neurobehavioral, metabolic, and immune dysfunctions through modulation of the gut microbiota. ### Competing Interest Statement The authors have declared no competing interest. Department of Biotechnology, https://ror.org/03tjsyq23, NABI CORE GRANT

Osteoarthritis (OA) is characterized by the deterioration of the articular cartilage in the joints, which activates the synthesis of pro-inflammatory cytokines, activating chondrocytes to release matrix metalloproteins (MMPs), thus exacerbating cartilage degradation. Risk factors for OA include age, gender, genetics, obesity, joint injuries, physical inactivity, metabolic disorders, etc. However, treatment options are still limited, with total joint replacement being the primary option for late-stage OA. Excessive and prolonged ER stress triggers apoptosis, whereas mild ER stress causes the induction of autophagy, offering the protection against apoptosis. However, the intricate interplay between ER stress, autophagy, and apoptosis in OA remains incompletely understood. So, in this study, we tried to elucidate the synergistic chondroprotective effect of ER stress inhibition and autophagy activation in ameliorating osteoarthritis. Our RT-qPCR results showed the significant upregulation of GRP78, PERK, ATF4, and CHOP genes of the UPR pathway along with the decrease in the expression of BECLIN 1, LC3, and ATG5 genes of autophagy in an IL-1β induced in vitro model of osteoarthritis. In line with these results, PERK inhibitor GSK2606414 caused significant downregulation of the PERK arm of the UPR pathway and rapamycin resulting in induction of autophagy in this in vitro OA model. Moreover, the combined usage of both the PERK arm inhibitor, GSK2606414, and autophagy activator, rapamycin, in IL-1β induced OA model significantly reduced the gene expression of COL1A1, RUNX2, and MMP13, and increased COL2A1 expression, which suggests prevention of OA progression. In summary, the study suggests the synergistic chondroprotective effect of PERK arm inhibition and autophagy activation in a mTOR dependent manner, which can be a promising strategy to ameliorate osteoarthritis progression.

INTRODUCTION:Enhancer of zeste homolog 2 (EZH2) is a histone methyltransferase that orchestrates gene expression via epigenetic and non-epigenetic mechanisms. EZH2 performs epigenetic functions by methylating histones and/or non-histone proteins and suppressing or activating target genes. Moreover, non-epigenetic functions involve dysregulation of target genes independent of histone methylation, thereby impacting multiple signaling pathways. AREAS COVERED:EZH2 has emerged as a pivotal player in the initiation of various cancers. EZH2 overexpression facilitated by H3K27me3 is the principal driver. However, the consequent dysregulation of target genes resulting from EZH2 overexpression has emerged as a secondary instigator of tumorigenesis, leading to metastasis and poor prognosis. Further complexity arises from somatic mutations in EZH2 and downstream target genes such as BRAF and RASSF1A. However, understanding its effects on endocrine tumors/cancers remains an underexplored with the potential to significantly enhance clinical outcomes and contribute to human health. Therefore, the present review focuses on the multifaceted functions of EZH2 and its pathophysiological mechanisms in tumor proliferation, with a specific emphasis on endocrine tumors. EXPERT OPINION:Investigating EZH2 mechanisms and targeting with inhibitors and drugs is an active area of research that could offer a promising avenue for treatment and a better understanding of molecular therapeutic interventions.

This study investigates proteomic differences between knee osteoarthritis (OA) and rheumatoid arthritis (RA) to identify protein signatures with potential diagnostic and therapeutic relevance. Using SWATH mass spectrometry, synovial fluid proteome from OA patients across Kellgren-Lawrence OA grades 2, 3, and 4 was analyzed and compared with RA patients. Out of 333 quantified proteins, 45 were differentially expressed, including aggrecan, versican, and inflammation-related proteins (e.g., CRP, APCS, S100A8, and SAA2). ELISA validation confirmed aggrecan, versican, and cartilage oligomeric matrix protein (COMP) as significantly altered proteins in OA compared to RA, along with distinct trends with OA progression and mirrored patterns in paired serum samples. ROC curve analysis highlighted COMP's strong diagnostic potential, with an AUC of 96%, 87.2%, and 85.2% for OA grades 2, 3, and 4 versus RA, respectively. COMP differentiated OA from RA at a synovial fluid concentration of < 3136 ng/mL, AUC of 92.1%, 89% sensitivity, and 82% specificity. Versican also demonstrated diagnostic utility, particularly in later OA stages. Gender-specific analysis revealed no differences for aggrecan and versican, while COMP levels were significantly higher in males. Simultaneously, a lower aggrecan, versican, and COMP levels were observed in OA (females) as compared to RA, potentially linked to estrogen decline with age and cartilage degradation. However, gender variability underscores the need for a larger, sex-balanced cohort study. Future studies could aim to account for validating COMP's diagnostic potential with healthy controls, demonstrating its reliability to characterize different OA grades.

Aim Osteoarthritis (OA) is the eleventh most disabling condition, with radiographic classification based on the Kellgren-Lawrence (KL) grading system. Early detection is critical to implement interventions to slow disease progression and improve patient outcomes. Proteomics, as a powerful strategy, could contribute to a better understanding of the disease pathophysiology and its early detection. Objectives The study aims to identify and confirm proteins associated with early detection and their role in the progression of knee OA. Methodology Synovial fluid (SF) and serum samples from the Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh, were categorized by KL classification and subjected to SWATHTM analysis in the discovery phase. Seven samples of each OA grade were analyzed. A mass dynamics tool was used for data analysis and visualization. Significant protein expression level was defined as -1≤ log2FC ≥1 with p-value <0.05, and ELISA was used for validation in a greater number of patients. Results 29 significantly modulated proteins were observed in osteoarthritis grade comparisons. Cathepsin G (CTSG) and angiotensinogen (AGT) were upregulated, whereas fumarylacetoacetase (FAH) and neural cell adhesion molecule 1 (NCAM1) were downregulated with radiographic disease progression, as validated by ELISA. CTSG, AGT, and NCAM1 showed good sensitivity and specificity in discriminating between early and late OA grades. Notably, serum and synovial fluid levels of AGT and NCAM1 exhibited significant correlation. Conclusion This is one of the first studies to comprehensively analyze proteins associated with OA progression. Additionally, the identified protein signatures have great potential for OA progression and differential diagnosis of early and late-stage OA.

HCC is the most common primary liver cancer, ranking as the sixth most prevalent cancer globally and the third leading cause of cancer-related deaths. Etiological factors include:chronic liver diseases driven by alcohol abuse, viral hepatitis, obesity, and metabolic disorders. Emerging evidence also suggests that gut microbiome alterations and subsequent immune and metabolic dysregulation contribute to the pathogenesis and progression of HCC. The gut-liver axis represents a dynamic interplay between the gastrointestinal tract and the liver, modulated by the gut microbiome, microbial metabolites, and immune responses. This bidirectional communication plays a pivotal role in maintaining metabolic homeostasis and immune surveillance, while its dysregulation is implicated in various pathologies, including hepatocellular carcinoma (HCC). The gut microbiome, through microbial dysbiosis and metabolite secretion, significantly influences the tumor microenvironment and immune evasion mechanisms in HCC. Perturbations in gut barrier function and Toll-like receptor 4 (TLR4) activation drive chronic inflammation, promoting tumor progression. Moreover, microbial metabolites such as short-chain fatty acids (SCFAs) and bile acids, modulate inflammatory and metabolic pathways, offering novel insights into disease pathogenesis and potential biomarkers. Therapeutic strategies, including probiotics, prebiotics, and immune checkpoint inhibitors demonstrate promise in reprogramming the gut microbiome and restoring immune balance in HCC management. This review explores the multifaceted roles of the gut-liver axis in pathogenesis, the contributions of the intra-tumoral microbiome, and the potential of microbial metabolites as therapeutic avenues. A deeper understanding of these interactions could pave the way for innovative, targeted interventions in liver cancer and other gut-liver axis-associated diseases.

Hepatocellular carcinoma represents a significant global cancer burden, ranking as the sixth most prevalent malignancy worldwide and constituting the third most frequent cause of cancer-related mortality. Folate and vitamin B12 play a crucial interdependent role in various physiological processes via their shared involvement in one-carbon metabolism. Most of the previous studies on patient cohorts have shown the association of levels of these vitamins with hepatocellular carcinoma progression and development. However, the combinatorial effect of folate and vitamin B12 on cancer hallmarks in HCC remains understudied. This study aims to determine the effects of an imbalance between folate and vitamin B12 in relation to HCC. HepG2 cells were cultured in media supplemented with varied levels of folic acid and vitamin B12 and were divided into five groups (BNFN, BDFO, FDBO, BNFO, and FNBO) (F—folic acid, B—vitamin B12, N—normal, D—deficient, O—over-supplemented), and various cancer hallmarks assays were performed. Cell migration assay revealed that the groups containing over-supplemented folic acid, irrespective of the levels of vitamin B12, showed increased migration, whereas migratory capacity was impeded in folic acid-deficient (FDBO) group. However, the invasion was increased in the groups FDBO and FNBO in association with a decrease in the percentage of viable cells and increased apoptosis. Interestingly, the number of spheres formed was considerably high in cells over- supplemented with folic acid (BDFO BNFO), concordant with the results found in the migration assay. Altogether, this suggests that an imbalance of low vitamin B12 and abundant folate aggravates cancer by enhancing cancer stem cell survival. Thus, these observations suggest that maintaining a proper balance between vitamin B12 and folate is essential for regulating cellular processes and potentially mitigating cancer progression.

In this manuscript, we comment on the article, which explores the anti-cancer effects of Calculus bovis (CB) in tumor biology. We highlight its potential, particularly in hepatocellular carcinoma (HCC), where it inhibits the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin pathways and induces apoptosis. CB contains compounds such as oleanolic acid and ursolic acid that target interleukin-6, mitogen-activated protein kinase 8, vascular endothelial growth factor, and caspase-3, offering anti-inflammatory and hepatoprotective benefits. The manuscript also discusses CB sativus (CBS), an artificial substitute, which has shown efficacy in reducing hepatic inflammation and oxidative stress in animal models. We emphasize the need for further research on the effects of CBS on the gut-liver axis and gut microbiota, and on targeting Wnt signaling and M2 tumor-associated macrophage as potential therapeutic strategies against HCC.

AbstractThe aberrant expression of placental imprinted genes due to epigenetic alterations during pregnancy can impact fetal development. We investigated the impact of dietary modification of low vitamin B12 with varying doses of folic acid on the epigenetic control of imprinted genes and fetal development using a transgenerational model of C57BL/6J mice. The animals were kept on four distinct dietary combinations based on low vitamin B12 levels and modulated folic acid, mated in the F0 generation within each group. In the F1 generation, each group of mice is split into two subgroups; the sustained group was kept on the same diet, while the transient group was fed a regular control diet. After mating, maternal placenta (F1) and fetal tissues (F2) were isolated on day 20 of gestation. We observed a generation-wise opposite promoter CpG methylation and gene expression trend of the two developmental genes Dlk1 and Grb10, with enhanced gene expression in both the sustained and transient experimental groups in F1 placentae. When fetal development characteristics and gene expression were correlated, there was a substantial negative association between placental weight and Dlk1 expression (r = − 0.49, p < 0.05) and between crown-rump length and Grb10 expression (r = − 0.501, p < 0.05) in fetuses of the F2 generation. Consistent with these results, we also found that H3K4me3 at the promoter level of these genes is negatively associated with all fetal growth parameters. Overall, our findings suggest that balancing vitamin B12 and folic acid levels is important for maintaining the transcriptional status of imprinted genes and fetal development.