Search Results (1,896)

The concentration of extracellular vesicles (EVs) in the peripheral blood of COVID-19 patients is increased. Nevertheless, their potential role in the transmission of infection remains unclear. This study was performed to determine whether EVs produced by the sub-genomic replicon system developed in Baby Hamster Kidney (BHK-21) cells could transfer SARS-CoV-2 replicon RNA, leading to the establishment of a viral replication system in human cells. Purified EVs from the SARS-CoV-2 sub-genomic replicon cell line BHK-21 were cultured with a naive human cell line. The success of EV-mediated transfer of SARS-CoV-2 replicon RNA and its productive replication was assessed using G-418 selection, a luciferase assay, immunostaining, and Western blot. We found that the A549 cell line cultured with EVs isolated from SARS-CoV-2 BHK-21 replicon cells developed G-418-resistant cell colonies. SARS-COV-2 RNA replication in A549 cells was confirmed by nano luciferase, Nsp1 protein. SARS-CoV-2 RNA replication causes massive morphological changes. Treatment of cells with the FDA-approved Paxlovid demonstrated a dose-dependent inhibition of viral replication. We isolated two human epithelial cell lines (gastrointestinal and neuroblastoma) and one vascular endothelial cell line that stably support high-level replication of SARS-CoV-2 sub-genomic RNA. Viral elimination did not revert the abnormal cellular shape, vesicle accumulation, syncytia formation, or EV release. Our study’s findings highlight the potential implications of EV-mediated transfer of replicon RNA to permissive cells. The replicon model is a valuable tool for studying virus-induced reversible and irreversible cellular reprogramming, as well as for testing novel therapeutic strategies for SARS-CoV-2. Full article

Retinal astrocytes reside mainly in the nerve fiber layer and are central to shaping retinal vessels and maintaining neurovascular balance. Derived from the optic nerve head, they spread across the inner retina to form a meshwork that both supports and instructs the emerging superficial vascular plexus. Immature astrocytes supply vascular endothelial growth factor-A(VEGF-A) to guide endothelial sprouting, while signals from growing vessels promote astrocyte maturation and strengthen the blood–retinal barrier. In disorders such as diabetic retinopathy and neovascular age-related macular degeneration, these cells show marked plasticity. Reactive astrogliosis can sustain VEGF and inflammation, favoring fragile, leaky neovessels, whereas alternative astrocyte states help reinforce barrier function and release anti-angiogenic factors. Located at the core of the neurovascular unit, astrocytes communicate continuously with endothelial cells, pericytes and neurons. This review integrates data from single-cell profiling and advanced imaging to outline astrocyte development, morphology and key signaling pathways (VEGF, PDGF, Wnt/Norrin, Eph/ephrin), and considers how tuning astrocyte polarization might be exploited to preserve retinal vascular integrity. Full article

Objective: This experiment aimed to explore the effects of water extract of Artemisia annua L. (WEAA) on growth performance, blood parameters, and intestinal-related indices in mutton sheep, so as to evaluate its potential as a natural growth promoter. Methods: The experiment was conducted using a completely randomized design. Thirty-two 3-month-old Dorper × Han mutton sheep were randomly assigned to 4 groups (n = 8). The control group was fed only the basal diet, while the other groups were fed the basal diet supplemented with, respectively, 500, 1000, and 1500 mg/kg WEAA. The adaptation period lasted 15 days, followed by a 60-day experimental period. Results: Results showed that dietary supplementation of WEAA significantly reduced average daily feed intake (ADFI) and feed-to-gain ratio (F:G) of mutton sheep, significantly improved the apparent digestibility of crude protein (CP) and phosphorus (P), and optimized blood biochemical indices, such as significantly increasing the concentrations of total protein (TP), albumin (ALB), high-density lipoprotein cholesterol (HDL-C), and glucose (GLU), while significantly decreasing blood urea nitrogen (BUN) level (p < 0.05). Additionally, WEAA significantly improved intestinal morphology by reducing the crypt depth (CD) of the duodenum, jejunum, and ileum, increasing jejunal villus height (VH), and elevating the villus-to-crypt ratio (VH/CD) across intestinal segments (p < 0.05). It also significantly enhanced the activity of intestinal digestive enzymes, including α-amylase and trypsin in the duodenum, lipase and chymotrypsin in the jejunum, and α-amylase and chymotrypsin in the ileum, with the 500 mg/kg and 1000 mg/kg WEAA groups reaching better activity (p < 0.05). Furthermore, WEAA supplementation significantly increased the counts of beneficial bacteria (Bifidobacteria and Lactobacilli) and decreased the count of harmful bacteria (Escherichia coli) in rectal fecal samples (p < 0.05). Notably, most of these beneficial effects were dosage-dependent, with overall optimal performance observed in the 1000 mg/kg WEAA group. Conclusion: In conclusion, supplementing the diet with 1000 mg/kg WEAA exerted significant positive effects on the feed efficiency, nutrient digestibility, and intestinal health status of mutton sheep. Full article

Background/Objectives: Branched-chain fatty acids (BCFAs) exhibit a range of biological activities; however, their limited natural abundance and high cost have constrained in vivo research. Lanolin represents a promising source for enriching BCFAs. Nevertheless, the in vivo application, safety, and dose-effect relationship of BCFAs derived from lanolin (BCFAs-DFL) remain unassessed. Methods: In this study, the acute toxicity in C57BL/6J mice was first evaluated for 7 days by a single oral administration of 5000 mg/kg BW of BCFAs-DFL. Subsequently, 40 mice were divided into four groups (control group, low dose of 100 mg/kg BW, medium dose of 300 mg/kg BW, and high dose of 600 mg/kg BW) and were continuously administered by gavage for 28 days to study the effects of BCFAs-DFL on the growth, blood biochemistry, intestinal morphology, and intestinal flora of the mice. Results: In the acute toxicity test, BCFAs-DFL exhibited no lethality or abnormalities in mice, indicating its non-toxic nature. Throughout the 28-day trial, mice in the medium- and high-dose groups experienced a notable decrease in average daily feed intake (p < 0.05), yet their weight gain remained unaffected (p > 0.05). Hemoglobin and hematocrit levels declined in the high-dose group (p < 0.05). Conversely, serum aspartate aminotransferase and total bilirubin levels escalated in the medium- and high-dose groups, while triglycerides and urea nitrogen levels decreased (p < 0.05). The serum’s total antioxidant capacity and immunoglobulin levels (IgA, IgG) rose in proportion to the dosage (p < 0.05). BCFAs-DFL notably enhanced the villus height of the jejunum and ileum in mice (p < 0.05). Gut microbiota analysis indicated no significant impact on overall α and β diversity. Conclusions: The 28-day intervention revealed that BCFAs-DFL can modulate feeding behavior, TG, T-AOC, and immunoglobulin levels in mice. Additionally, it promotes the development of intestinal villi. Based on various indicators, a dosage of 100 mg/kg BW effectively induces beneficial metabolic regulation, such as the reduction of triglycerides, without causing a burden on liver metabolism. This dosage may represent a more suitable application for potential use. Full article

This study investigated the effects of dietary probiotic supplementation and stocking density on the growth performance, carcass traits, immunity, blood biochemical parameters, and ileal histomorphology of broiler chickens. A total of five hundred ten 1-day-old unsexed broiler chicks (Cobb 39) were allocated to three probiotic levels (0%, 0.1%, 0.2%) and two stocking densities (low vs. high). Results indicated that stocking density significantly influenced body weight from the third week onward, with birds reared under low density showing higher weight and better feed-to-gain ratio. Probiotic supplementation did not significantly affect weekly body weight, feed intake, or mortality, although mortality tended to be lower in probiotic-fed groups. Carcass traits and lymphoid organ indices were largely unaffected by treatments, except for a higher heart percentage in low-density birds. Cell-mediated immunity was enhanced under low stocking density, and probiotic supplementation at 0.2% increased the immune response at 48 h post-challenge. Blood biochemical analysis revealed significant effects of stocking density on total protein, globulin, and triglycerides, while probiotics reduced total lipid and LDL levels. Ileal histomorphology was significantly improved by probiotics, with increased villus height, crypt depth, and villus-to-crypt ratio. Similarly, low stocking density further enhanced these parameters. Overall, probiotic supplementation, particularly at 0.1%, combined with low stocking density, positively influenced gut morphology and immune responses, contributing to improved broiler health and performance. Full article

Objective: This study compares the brain protective effects of L-borneolum and its main components (a combined application of L-borneol and L-camphor) on the rat model of middle cerebral artery occlusion/reperfusion (MCAO/R). It also makes clear the intrinsic regulatory mechanisms that link the neuroprotective effects of these compounds on IS to the blood-brain barrier (BBB), based on network pharmacology predictions. Furthermore, the study investigates the relationship between these compounds and the Major Facilitator Superfamily Domain-containing Protein 2A (MFSD2A)/Caveolin-1 (Cav-1) signaling axis. Methods: The MCAO/R model in rats was established to evaluate the therapeutic effect of L-borneolum (200 mg/kg) and its main components combination of L-borneol and L-camphor (6:4 ratio, 200 mg/kg). Neurological scores, 2,3,5-triphenyl tetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, and Nissl staining were performed to evaluate the neurological damage in the rats. Cerebral blood flow Doppler was applied to monitor the cerebral blood flow changes. Immunofluorescence analysis of albumin leakage and transmission electron microscopy (TEM) were conducted to evaluate blood-brain barrier (BBB) integrity. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the optimal drug concentration. Trans-epithelial electrical resistance (TEER) and horseradish peroxidase (HRP) assays were employed to confirm the successful establishment of an in vitro BBB co-culture model. Network pharmacology was utilized to predict the biological processes, molecular functions, and cellular components involved in the treatment of ischemic stroke (IS) by the main components of L-borneolum (L-borneol and L-camphor). Finally, immunofluorescence, real-time fluorescent quantitative PCR (RT-qPCR) and western blot analyses were performed to detect the expression of Major Facilitator Superfamily Domain Containing 2A (MFSD2A), caveolin-1 (CAV-1), sterol regulatory element-binding protein 1 (SREBP1) in brain tissue and hCMEC/D3 cells. Results: Network pharmacology prediction indicated that L-borneolum and its main components (L-borneol and L-camphor) in the treatment of IS are likely associated with vesicle transport and neuroprotection. Treatment of IS with L-borneolum and its main components significantly decreased neurological function scores and cerebral infarction area, while alleviating pathological morphological changes and increasing the number of Nissl bodies in the hippocampus. Additionally, it improved cerebral blood flow, reduced albumin leakage, and decreased vesicle counts in the brain. The trans-epithelial electrical resistance (TEER) of the co-culture model stabilized on the fifth day after co-culture, and the permeability to horseradish peroxidase (HRP) in the co-culture model was significantly lower than that of the blank chamber at this time. RT-qPCR and Western blot results demonstrated that, compared to the model group, the expression of SREBP1 and MFSD2A significantly increased, while the expression of Cav-1 decreased. Conclusions: L-borneolum and its main components combination (L-borneol/L-camphor, 6:4 ratio) may exert a protective effect in rats with IS by improving BBB transport function through modulation of the MFSD2A/Cav-1 signaling pathway. Full article

Background: There exists some inconsistent evidence on the relationship between altered cardiac morphology, its function, and frailty. Therefore, this study aimed to assess the associations among frailty, lean body mass, central arterial stiffness, and cardiac structure and geometry in older people with a normal ejection fraction. Methods: A total of 205 patients >65 years were enrolled into this ancillary analysis of the FRAPICA study and were assessed for frailty with the Fried phenotype scale. Left ventricular dimensions and geometry were assessed with two-dimensional echocardiography. Fat-free mass was measured using three-site skinfold method. Parametric and non-parametric statistics and analysis of covariance were used for statistical calculations. Results: Frail patients were older and women comprised the majority of the frail group. Frail men and women had comparable weight, height, fat-free mass, blood pressure, central blood pressure, and carotid–femoral pulse wave velocity to their non-frail counterparts. There was a linear correlation between the sum of frailty criteria and left ventricular end-diastolic diameter (Spearman R = −0.17; p < 0.05) and relative wall thickness (Spearman R = 0.23; p < 0.05). In the analysis of covariance, frailty and gender were independently associated with left ventricular mass (gender: β of −0.37 and 95% CI of −0.50–−0.24 at p < 0.001), the left ventricular mass index (gender: β of −0.23 and 95% CI of −0.37–−0.09 at p < 0.001), and relative wall thickness (frailty: β of −0.15 and 95% CI of −0.29–−0.01 at p < 0.05; gender: β of 0.23 and 95% CI of 0.09–0.36 at p < 0.01). Frailty was associated with a shift in heart remodeling toward concentric remodeling/hypertrophy. Conclusions: Frailty is independently associated with thickening of the left ventricular walls and a diminished left ventricular end-diastolic diameter, which are features of concentric remodeling or hypertrophy. This association appears to be more pronounced in women. Such adverse cardiac remodeling may represent another phenotypic feature linked to frailty according to the phenotype frailty criteria. Full article

Microscopic examination of Giemsa-stained thick blood smears remains the reference standard for malaria diagnosis, but it requires specialized personnel and is difficult to scale in resource-limited settings. We present a lightweight, smartphone-based system for automatic detection of Plasmodium parasites in thick smears captured with mobile phones attached to a conventional microscope. We built a clinically validated dataset of 400 slides from Loreto, Peru, consisting of 8625 images acquired with three smartphone models and 54,531 annotated instances of Plasmodium vivax and P. falciparum across eight morphologic classes. The workflow includes YOLOv11n-based visual-field segmentation, rescaling, tiling into 640 × 640 patches, data augmentation, and parasite detection. Four lightweight detectors were evaluated; YOLOv11n achieved the best trade-off, with an F1-score of 0.938 and an overall accuracy of 90.92% on the test subset. For diagnostic interpretability, performance was also assessed at the visual-field level by grouping detections into Vivax, Falciparum, Mixed, and Background. On a high-end smartphone (Samsung Galaxy S24 Ultra), the deployed YOLOv11n model achieved 110.9 ms latency per 640 × 640 inference (9.02 FPS). Full article

Glioblastoma (GBM) is a highly aggressive brain tumor characterized by invasive growth, intrinsic drug resistance, and the presence of the blood–brain barrier. All of these features make treatment extremely challenging and underscore the need for developing effective combination strategies and advanced drug delivery systems. This study aimed to develop a bovine serum albumin (BSA) nanoparticle (NP)-based delivery system to overcome the poor bioavailability and pharmacokinetic limitations of two potent anti-tumor agents, all-trans retinoic acid (ATRA) and curcumin (CURC), and to evaluate their antitumor activity in U87-MG GBM cells. Drug-free and ATRA/CURC-loaded BSA-NPs were synthesized using an optimized desolvation method and characterized in terms of particle size, polydispersity index, morphology, drug encapsulation efficiency, and release behavior. The cytotoxic, anti-migratory, and pro-apoptotic effects of the NPs on U87-MG GBM cells were assessed using real-time proliferation and migration assays and Annexin V/PI staining followed by flow cytometry. Collectively, the findings indicated that the co-delivery of ATRA and CURC using BSA-NPs showed enhanced antiproliferative, antimigratory, and pro-apoptotic effects. With its controlled release profile, high loading capacity, and favorable nanoscale dimensions, the ATRA-CURC-BSA–NP system represents a promising nanoplatform for GBM therapy that warrants further in vivo investigation. To the best of our knowledge, this is the first study demonstrating the inhibition of glioblastoma cell growth through the co-delivery of all-trans retinoic acid and curcumin using a bovine serum albumin-based nanoparticle system. Full article

The forest fly (Hippobosca equina) is an obligate haematophagous dipteran insect (order Diptera) that primarily infests horses and may contribute to the circulation of vector-borne pathogens. This study aimed to investigate the presence of Anaplasma phagocytophilum, Borrelia burgdorferi s.l., Babesia caballi, and Theileria equi, important vector-borne pathogens of equids, in forest flies collected from horses in endemic areas of Spain. A total of 170 forest flies were collected from 39 equids across four geographical regions in Spain (Segovia, Madrid, Toledo, and Menorca) and blood samples were collected from 27 of these horses. All flies were morphologically and molecularly identified as H. equina, and DNA extracted from flies and equine blood was screened using multiplex real-time and nested PCR, followed by sequencing and phylogenetic analysis. Neither flies nor horses tested positive for A. phagocytophilum, whereas one fly was positive for B. burgdorferi s.l. (0.6%). In contrast, T. equi and B. caballi DNA were detected in 11.2% and 1.2% of flies, respectively, and all positive flies were collected from horses positive for equine piroplasmosis (T. equi/B. caballi infection), with identical 18S rRNA sequences between hosts and flies. Nested PCR showed a higher detection rate than real-time PCR for the detection of these piroplasms in flies and blood samples. These findings provide the first molecular evidence of EP pathogens in H. equina and support further investigation into the epidemiological importance of forest flies in equine pathogen surveillance. Full article

Fibrous polycaprolactone-based composite materials with the addition of hardystonite (1, 3, and 5 wt.%) were developed using the electrospinning method. The obtained PCL and PCL-HT nonwovens were evaluated in terms of their physiochemical properties (SEM, EDS, BET, and zeta potential). Furthermore, the antioxidant potential [measured by thiobarbituric acid reactive substance (TBARS) levels], blood plasma coagulation parameters, and cyto- and genotoxicity towards PBM and Hs68 cells were assessed to determine the biochemical activity of the composites. The conducted experiments confirmed that hardystonite was successfully incorporated into the PCL matrix. No substantial changes in the fibres’ surface morphology and the structure of the composites were observed. Similarly, the specific surface area, total pore volume, and average pore size did not change significantly. The addition of hardystonite to the polymer solution resulted in a shift in zeta potential toward less negative values. With regard to plasma coagulation parameters, no significant changes were observed in the aPTT, PT, or TT, likely due to the counterbalancing effect of Zn²⁺ and Ca²⁺ ions. Furthermore, the PCL-HT composites exhibited a lowered TBARS level, suggesting antioxidant properties, which could be attributed to the presence of zinc in hardystonite. The PCL and PCL-HT composites demonstrated no cytotoxic or genotoxic effects on the tested blood or skin cell types, suggesting their safety. Full article

A facile and single-step synthesis of poly(L-Cysteine) (p(L-Cys)) particles through microemulsion polymerization using tetrakis(hydroxymethyl) phosphonium chloride (THPC) as crosslinker is accomplished for the first time. The L-Cys:THPC ratio in p(L-Cys) particles was calculated as 80:20% (by weight) with elemental analyses, and the generation of p(L-Cys) particles was confirmed. SEM imaging revealed a popcorn-like morphology of the p(L-Cys) particles with a 1–20 µm particle size range. The isoelectric point of p(L-Cys) particles was determined at pH 1.15 via zeta potential measurements. The hydrolytic degradation of p(L-Cys) particles was determined as about 85% within 3 h (by weight). The p(L-Cys) particles displayed excellent blood compatibility with a hemolysis % ratio of <2.3% and a blood clotting index of 95% at 1 mg/mL concentration. Moreover, cell compatibility tests up to 50 mg/mL against L929 fibroblast cells exhibited about 90% cell viability for p(L-Cys) particles versus 58% for L-Cys molecule. The antimicrobial efficacy of the L-Cys molecules was notably enhanced in p(L-Cys) particles, exhibiting a 5-fold reduction in minimal bactericidal concentration (MBC) values against E. coli (Gram-negative, ATCC 8739) and a 2-fold reduction against S. aureus (Gram-positive, ATCC 6538). Additionally, the antioxidant capacity of p(L-Cys) particles was retained somewhat, measured as 0.14 ± 0.01 µM versus 2.25 ± 0.03 µM Trolox equivalent/g for L-Cys. Therefore, p(L-Cys) particles are versatile and offer a unique avenue for immense biomedical use. Full article

Ticks and tick-borne diseases pose a significant threat to public health. The Tumen River Basin is located at the junction of China, North Korea and Russia, whose warm climate and favorable ecological environment are suitable for the growth and reproduction of ticks. At the same time, the cattle industry in this region is highly developed, with cattle serving as the primary economic source for the area. This study performed an epidemiological investigation and analysis of pathogens carried by ticks and cattle in the Tumen River basin. A total of 913 ticks and 247 bovine blood samples were collected from seven cities primarily focused on cattle farming in the Tumen River basin. Morphological and molecular biological identification of ticks was carried out to determine the distribution of ticks and their pathogens in the region. Through the detection of pathogens carried by cattle blood samples in the surrounding area, the correlation with tick distribution was confirmed. The species and distribution of ticks of different genders and in different collection environments, and the infection of pathogens in bovine blood samples were statistically analyzed. The results showed that the 913 ticks had 5 species, including Haemaphysalis longicornis, Haemaphysalis concinna, Haemaphysalis japonica, Dermacentor silvarum and Ixodes persulcatus. Three pathogens, Babesia ovata, Theileria orientalis and Theileria sinensis, were detected in the blood samples of vector ticks and cattle. These results provide a theoretical basis for the prevention and control of ticks and tick-borne diseases in the Tumen River basin. Full article

Background and Objectives: Neutrophil extracellular traps (NETs) have been linked to hypercoagulability, immunothrombosis, and organ injury in COVID-19. Digital morphology of peripheral blood smears enables the identification of NET-compatible appearances (NET-like) in circulation, and associations between NET-like derived indices and clinical outcomes have been reported. However, evidence at hospital admission that relates smear NET-like burden to systemic inflammation and clinical severity remains limited. We therefore aimed to compare the burden of NET-like structures on admission smears according to disease severity and systemic inflammatory markers. Materials and Methods: We included 50 consecutively enrolled adults hospitalized for COVID-19; samples were obtained within 24 h of admission. Severity was defined by the World Health Organization Clinical Progression Scale and grouped as moderate or severe. C-reactive protein (CRP), ferritin, and complete blood counts were measured; the neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) were calculated. Digital morphology assessed 200 leukocytes per patient; the presence of morphological abnormalities, including NET-like events per patient, was recorded. We additionally quantified NET-like events per 100 white blood cells (NET-like/100 WBC) and the neutrophil extracellular trap–segmented neutrophil ratio (NNSR). Results: At admission, CRP, ferritin, NLR, and PLR of patients were above method-specific reference intervals. NET-like events were identified in 66% of patients. NET-like/100 WBC correlated positively with NLR (r = 0.312; p < 0.05). Patients with severe COVID-19 had higher NET-like/100 WBC than those with moderate disease (5.8 ± 7.34 vs. 14.14 ± 15.12; p = 0.0294). Conclusions: Digital morphological identification of NET-like structures on peripheral blood smears is frequent at admission and is associated with systemic inflammatory burden and with greater COVID-19 severity. These findings support the potential complementary value of reporting NET-like events for initial risk stratification in the clinical laboratory. Full article

Objectives: To systematically analyze the expression profiles of long non-coding RNAs (lncRNAs) in serum-derived exosomes from patients with age-related hearing loss (ARHL), and to further identify key regulatory lncRNAs involved in the pathogenesis and progression of ARHL. Methods: Peripheral blood samples were collected from patients with ARHL and age-matched normal-hearing controls. Serum was separated and exosomes were extracted. The exosomes were identified by nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and Western blot. Subsequently, total RNA was extracted from the purified exosomes for lncRNA transcriptome sequencing. Based on the sequencing results, we identified differentially expressed lncRNAs and mRNAs and conducted multi-dimensional functional analysis, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Reactome pathway database (Reactome), and Disease Ontology (DO). Finally, four key mRNAs (THAP2, ZNF225, MED12, and RNF141) and four differentially expressed lncRNAs (DE-lncRNAs), namely MSTRG.150961.7, ENSG00000273015, MSTRG.336598.1, and ENSG00000273493, were experimentally verified by quantitative real-time polymerase chain reaction (RT-qPCR) technology. Results: Exosomes were successfully isolated from serum and confirmed by particle size, morphological examination, and the expression of exosome-labeled proteins. A total of 2874 DE-lncRNAs were identified, among which 988 were downregulated and 1886 were upregulated. Similarly, 2132 DE-mRNAs were detected, among which 882 were downregulated and 1250 were upregulated. GO analysis revealed significant enrichment in biological processes such as “phospholipid binding”, “phosphatidylinositol binding”, “phosphatase binding”, “phosphatidylinositol bisphosphate binding”, “phosphatidylinositol-4,5-bisphosphate binding”, “phosphatidylinositol-3,5-bisphosphate phosphatase activity”. KEGG is significantly enriched in signaling pathways including “Wnt signaling pathway”, “Hippo signaling pathway”, “Cushing syndrome”, and “Nucleocytoplasmic transport”. The functional annotations of Reactome were significantly enriched in biomolecular pathways including “tRNA processing”, “Cellular response to heat stress”, “Extra-nuclear estrogen signaling”, “Metabolism of non-coding RNA”, and “CTNNB1 T41 mutants aren’t phosphorylated”. DO is significantly enriched in diseases or pathological conditions such as “hepatitis”, “bacterial infectious disease”, “cystic fibrosis”, and “vasculitis”. Conclusions:THAP2, ZNF225, MED12, and RNF141 may serve as potential candidate biomarker for ARHL. Additionally, lncRNA MSTRG.150961.7, lncRNA MSTRG.336598.1, and lncRNA ENSG00000273493 may play significant roles in the pathogenesis of this condition. Full article