Home Life Sciences Role of DZ2002 in reducing corneal graft rejection in rats by influencing Th17 activation via inhibition of the PI3K/AKT pathway and downregulation of TRAF1
Article Open Access

Role of DZ2002 in reducing corneal graft rejection in rats by influencing Th17 activation via inhibition of the PI3K/AKT pathway and downregulation of TRAF1

  • , , , , , , and ORCID logo EMAIL logo
Published/Copyright: January 1, 2026
Download Article (PDF)
Open Life Sciences
From the journal Open Life Sciences Volume 20 Issue 1

Abstract

To investigate the mechanism of DZ2002, a reversible type III S-adenosyl-l-homocysteine hydrolase (SAHH) inhibitor, in alleviating corneal graft rejection in rats through the downregulation of TRAF1 and modulation of Th17 differentiation, so as to provide a new therapeutic strategy for corneal graft rejection. Rat models of corneal graft rejection were constructed and treated with DZ2002 eye drops. Metabolomics and transcriptomics sequencing analyses were used to identify the key metabolite, homocysteine, and its associated gene, TRAF1. Histological assessments were conducted to evaluate the effect of DZ2002 on corneal graft rejection. Additionally, the underlying mechanism of action was explored through Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Homocysteine emerged as a key metabolite involved in corneal graft rejection, and DZ2002 could significantly decrease the rejection index and reduce the opacity and edema of corneal grafts. The effect of DZ2002 in alleviating rejection was attributed to the downregulation of TRAF1 expression, inhibition of the PI3K/AKT pathway, decreased expression of chemokine CCL5 in the aqueous humor, and suppression of the differentiation of T cells into Th17 cells. DZ2002 was effective in reducing corneal graft rejection in rats by downregulating TRAF1 and modulating Th17 differentiation.

Keywords: corneal transplantation; DZ2002; homocysteine; PI3K/AKT signaling pathway; rejection; TRAF1

1 Introduction

Corneal blindness is a type of ocular disease that leads to corneal dysfunction and loss of vision stemming from a variety of etiologies, including keratitis, corneal degeneration, and corneal chemical injury [1]. Recent statistics indicate a substantial global burden, with about 217 million people suffering from moderate to severe vision impairment and an estimated 36 million individuals affected by blindness [2]. Notably, China ranks among the countries with the highest prevalence of blindness, with an estimated six to nine million people with bilateral blindness and about four million patients with corneal blindness who are in urgent need of surgery, with the incidence of corneal blindness escalating at a rate of 10 % per year [3].

Corneal transplantation remains the primary recourse for patients with corneal blindness in an attempt to restore vision and achieve better outcomes [4], 5]. However, despite the presence of anterior chamber immune deviations in the cornea, the 10–50 % incidence of allogeneic corneal transplantation rejection can lead to poor vision correction or even a relapse of blindness [6]. Corneal graft rejection is a complex immune response involving multiple factors, and its mechanism remains poorly understood. At present, glucocorticoids and immunosuppressants used in clinical practice can improve corneal graft rejection, but fail to completely and effectively control rejection or avoid side effects including hypertension, osteoporosis, and glaucoma blindness caused by long-term medication regimens. Therefore, exploring the mechanisms of corneal graft rejection and identifying possible treatment measures to prevent rejection assumes significance.

Protein metabolism during rejection is considerably different from that observed in a normal cornea with long-term inflammation resulting in fibrosis of the corneal tissue, thereby affecting corneal transparency. Metabolomics is an emerging field in the study of corneal diseases that provides new perspectives and analytical tools for uncovering the pathogenesis, diagnosis, and treatment of corneal diseases. The aim of the current study was to explore the mechanism of the metabolite homocysteine (Hcy) in the form of DZ2002, a reversible S-adenosyl-l-homocysteine hydrolase (SAHH) inhibitor, in corneal graft rejection. This can generate insights related to the modulation of corneal metabolism and facilitate the development of new therapeutic drugs for corneal graft rejection.

2 Materials and methods

2.1 Experimental animals

SPF Wistar female rats (weighing 180–200 g) and SD female rats (weighing 180–200 g) were purchased from Hunan SJA Laboratory Animal Co. (Certificate No. 43004700043639, China), license number SCXK (Beijing) 2019-0010. The rats were randomly divided into two groups: the blank control group (which received no treatment) and the corneal graft rejection group (where a graft rejection model of allogeneic penetrating keratoplasty was established). The right eyes were chosen for surgery, while the left eyes remained untreated so that the postoperative feeding of the rats would not be affected. The use and feeding of the experimental animals were in accordance with the Regulations for the Administration of Affairs Concerning Experimental Animals approved by the State Science and Technology Commission.

Ethical approval: The research related to animal use has been complied with all the relevant national regulations and institutional policies for the care and use of animals, and has been approved by the Ethics Committee of Affiliated Calmette Hospital of Kunming Medical University (No.YLS2023-67).

2.2 Establishment of the penetrating keratoplasty graft rejection model

Initially, tropicamide eye drops were administered to both the donor (Wistar female rats) and recipient (SD female rats) eyes three times, with each application lasting 5 min. An overdose of anesthesia was then induced via an intraperitoneal injection of 0.5 ml of equal amounts of ketamine and diazepam. Subsequently, corneal grafts were procured from both eyes of Wistar rats (φ = 4.25 mm) and preserved in Optisol liquid. SD female rats served as recipients and were administered oxybuprocaine hydrochloride eye drops once.

The recipient cornea was incised at the center with a trephine (φ = 4 mm), reaching a depth of 1/2 of the corneal thickness. A 15° puncture knife was used to create a wedge in the anterior chamber, and an appropriate amount of Healon was injected to protect the corneal endothelium and maintain the anterior chamber. The corneal sheet was dissected with VANNAS scissors along the incision, and the donor corneal graft was positioned in the implant bed. Eight intermittent stitches with 10-0 micro-sutures were made, leaving the suture knot exposed. The anterior chamber was rinsed with 0.9 % normal saline to replace the Healon. The needle was flushed, and sterile air was injected to maintain the anterior chamber. The eyelids were sutured with 6-0 sutures after applying tobramycin eye ointment to the conjunctival sac.

Post-surgery, tobramycin ophthalmic ointment was applied once a day to prevent infection. Daily observations of the graft were conducted using a stereomicroscope from the first postoperative day, and three aspects, namely, graft opacity, edema, and angiogenesis, were assessed [7]. Scores of these three parameters were summed up to obtain the rejection index (RI) each day, with rejection defined as RI ≥ 6. The observation endpoint was four weeks post-surgery, after which the rats were euthanized.

Grouping:

  1. Control group (subconjunctival injection of physiological saline):Control

  2. Corneal transplant rejection model group (subconjunctival injection of physiological saline):Allo

  3. Homocysteine inhibitor group (DZ20024 times/day after transplantation in rats):Allo + DZ2002

  4. TRAF1 knockout group (transplantation of rat subconjunctival injection of TRAF1 knockout adenovirus)

  5. Homocysteine inhibitor + TRAF1 overexpression group (after transplantation in rats, point DZ2002 and subconjunctival injection of TRAF1 overexpressing adenovirus):Allo + DZ2002 + oe-TRAF1

2.3 Hematoxylin and eosin (HE) and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining

Tissue samples were collected and immersed in 4 % paraformaldehyde overnight at 4 °C. Subsequently, they were paraffin-embedded after ethanol gradient dehydration (two days each with 70 % ethanol, 96 % ethanol, and 100 % ethanol, respectively). Sections with a thickness of 5 μm were then obtained for HE and TUNEL staining.

2.4 Metabolomics sequencing for identifying the key metabolites involved in corneal graft rejection

2.4.1 Sample pretreatment and metabolite extraction

Serum samples, including those for sample preparation quality control (QC), from the corneal graft rejection group and the normal group, with six replicates per group, were analyzed using ultra-performance liquid chromatography-mass spectroscopy (UPLC-MS) after appropriate sample treatment. Metabolites were isolated and detected using the Waters UPLC I-Class Plus (Waters, USA) along with a tandem Q Exactive High Resolution Mass Spectrometer (Thermo Fisher Scientific, USA).

2.4.2 Data preprocessing and quality control (QC)

The results obtained from Compound Discoverer were imported into MetaX for data preprocessing. This preprocessing entailed normalization of the data using the Probabilistic Quotient Normalization (PQN) method to obtain the relative peak area and correction of batch effects utilizing the quality control-based robust LOESS signal correction (QC-RLSC). Compounds with a coefficient of variation (CV) of relative peak area >30 % were eliminated from all QC samples.

2.4.3 Screening of differential metabolites

The screening of differential metabolites involved univariate and multivariate analyses to identify variations between the two biological groups. The overall differences between the two groups were assessed using principal component analysis (PCA) and partial least squares discriminant analysis (PLSDA). Subsequently, the variable important for the projection (VIP) values of metabolites were analyzed through orthogonal partial least squares discriminant analysis (OPLSDA), and the VIP value of PLSDA was adopted if OPLSDA was prone to overfitting. Differential metabolites were screened using fold change and P values in univariate analysis, and volcano diagrams were plotted. In this study, differential metabolites were identified as per the following conditions: 1) VIP ≥ 1 in the OPLS-DA model; 2) fold change ≥1.2 or ≤0.83; and 3) P value < 0.05.

2.5 Surface plasmon resonance (SPR) assay

2.5.1 TRAF1 immobilization

The running buffer was PBST.

The carboxylated surfaces of flow cells Fc2 and Fc1 were activated with 0.4 M EDC and 0.1 M NHS at an injection volume of 170 μl and flow rate of 20 μl/min. TRAF1 protein was then coupled to Fc2 using 100 μg/ml protein in sodium acetate buffer (pH 4.5), with an injection volume of 170 μl and flow rate of 20 μl/min. Finally, both Fc2 and Fc1 were blocked with 1 M ethanolamine, using an injection volume of 170 μl and flow rate of 20 μl/min.

2.5.2 TRAF1–L-homocysteine interaction

The running buffer was PBST. The analyte was dissolved in PBST, and the interaction was analyzed using the Fc2–Fc1 double-referenced channel.

A series of L-homocysteine concentrations (0, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, and 500 μM) were injected sequentially, and signal responses were recorded in real time.

2.6 Transcriptomics sequencing for identifying the key genes associated with Hcy-induced corneal graft rejection

2.6.1 Experimental procedure

RNA was isolated and purified using TRIzol (Thermo Fisher, 15596018) and tested for integrity using the Bioanalyzer 2100 (Agilent, CA, USA). This was followed by paired-end sequencing performed with the Illumina Novaseq™ 6000 (LC-Bio Technologies Co., Ltd., Hangzhou, China) in PE150 mode.

2.6.2 Data quality control

The raw sequencing data were preprocessed and filtered. Genes were screened based on both fold change (FC) and significance level. Specifically, an FC value ≥ 2 or FC ≤ 0.5 (i.e., the absolute value of log2FC ≥ 1) and the q value < 0.05 (|log2fc| ≥ 1 and q < 0.05) were used as the threshold criteria. In cases where multiple groups were compared without FC, genes with a q value < 0.05 were considered genes showing statistically significant differences between multiple groups. Subsequently, clean data were obtained after filtering. These data were compared with the reference genome to obtain comprehensive alignment information.

Additionally, using the gene location information specified in the genome annotation file (gtf), the read data between the sequencing data and reference genome as well as the regional distribution data between the sequencing data and reference genomes were compared. (Genome: ftp://ftp.ensembl.org/pub/release-101/fasta/mus_musculus/dna/).

2.7 RT-PCR for validating the mRNA expression of differentially expressed genes

Total RNA was extracted manually, ensuring a purity of 1.9–2.0. cDNA was synthesized using reverse transcription and stored in a 4 °C freezer for subsequent use. Quantitative assessment of the RNA of samples was conducted, and the relative expression of mRNA was determined based on the cycle threshold (Ct) values.

2.8 Enzyme-linked immunosorbent assay (ELISA) for detecting the expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), and transforming growth factor-β (TGF-β)

The procedure outlined in the enzyme-linked immunosorbent assay (ELISA) kit was meticulously followed for detecting the expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), and transforming growth factor-β (TGF-β), and the absorbance was measured at 450 nm using a microplate reader (BioTek, Elx800).

2.9 Western blotting (WB) for detecting protein expression levels

Samples were lysed on ice for 10 min and centrifuged at 14,000 g at 4 °C for 15 min. Protein concentration was measured with the BCA Protein Quantification Kit. A total of 80 μl of protein and 20 μl of 5× protein loading buffer were mixed well and boiled in a water bath for 5 min. SDS-PAGE electrophoresis was then performed. Upon completion of electrophoresis, the proteins were transferred onto a PVDF membrane. The membrane strips were submerged in 1 × TBST and subsequently placed in a blocking solution containing 5 % skimmed milk powder on a shaker at room temperature for 40 min to block non-specific binding sites. Primary antibodies targeting specific proteins (TRAF1, AKT, P-AKT, PI3K, and P-PI3K) were incubated at 4 °C overnight. Secondary antibodies were added and allowed to incubate for 40 min. The membrane strips were then developed and photographed to visualize the protein bands.

2.10 Immunohistochemistry (IHC) staining detecting the vascular endothelial growth factor (VEGF) positive rate

Tissue specimens were initially fixed overnight in 4 % paraformaldehyde, paraffin-embedded after ethanol gradient dehydration, and stained for immunohistochemistry in 5 μm-thick sections. Primary antibodies used for immunohistochemistry included rat Vascular Endothelial Growth Factor (VEGF) antibody (Affinity, AF5131) at a dilution of 1:50.

2.11 Flow cytometry analysis of the ratio of regulatory T (Treg) and T helper 17 (Th17) cells grouping

  1. CD4 + T cell group: blank control group.

  2. HCEC-OE-NC + CD4 + T cell group: Co culture of overexpressing control human corneal epithelial cells (HCEC) and CD4 + T cells.

  3. HCEC-OE-TRAF1 + CD4 + T cell group: Co culture of HCEC cells overexpressing TRAF1 and CD4 + T cells.

  4. HCEC-OE-NC + DZ2002 + CD4 + T cell group: Co culture with CD4 + T cells after adding inhibitor DZ2002 to overexpressing control HCEC cells.

  5. HCEC-OE-TRAF1 + DZ2002 + CD4 + T cell group: HCEC cells overexpressing TRAF1 were co cultured with CD4 + T cells after adding inhibitor DZ2002.

Peripheral blood samples were collected from healthy volunteers to obtain peripheral blood mononuclear cells (PBMCs) by centrifugation. The PBMCs were washed once with phosphate buffer solution (PBS) and incubated for 30 min. The cells were then incubated with 2 μL of Brefeldin A (5 mg/ml) for 4 h to facilitate stimulation. Then cells were washed once with PBS and incubated with FITC-CD at 4 °C for 15 min in the dark. After 30 min of incubation with a permeabilization agent, the cells were treated with PE-IL-17, Perep/Cy5.5-IL4, and APC-IFN-γ monoclonal antibodies at 4 °C for 15 min. Finally, the cells were washed once with PBS, resuspended, and subjected to flow cytometry analysis to determine the proportion of Regulatory T (Treg) and T helper 17 (Th17) cells. Stained samples were acquired on a BD FACSCalibur flow cytometer, collecting 10,000 events per sample, and the results were analyzed using FlowJo software to quantify cellular expression.

2.12 Statistical analysis

Prism graphpad was used for statistical analysis. Experimental data were represented as the mean ± standard error (mean ± SEM). The t-test was used for comparison of measurement data between groups, and one-way analysis of variance (ANOVA) was used for comparison of measurement data between multiple groups. A P value of <0.05 was considered to indicate a statistically significant difference; a P value of <0.01 was considered to indicate high statistical significance; and a P value of <0.001 was considered to indicate a very significant statistical difference.

3 Results

3.1 Metabolomics sequencing revealed that Hcy is a key metabolite involved in corneal graft rejection

To study the underlying mechanisms of rejection following a corneal transplantation, we initially established a rat corneal graft rejection model. The corneal graft was observed under a stereomicroscope. After four weeks of transplantation, the graft opacity, stromal edema, and RI scores were significantly increased (Figure 1A and B). HE staining showed that, compared with the cornea of the normal group, the grafts of the rejection group were thicker due to edema, with noticeable infiltration of inflammation cells, indicative of a more pronounced inflammatory response accompanying the rejection of the surgical eye (Figure 1C).

Figure 1: Establishment of the rat corneal transplantation rejection model. (A) Observation of corneal graft rejection under a stereomicroscope. (B) Statistical scoring of rejection severity. (C) H & E staining showing pathological changes in corneal tissue at 4 weeks post-transplantation (200 × magnification).
Figure 1:

Establishment of the rat corneal transplantation rejection model. (A) Observation of corneal graft rejection under a stereomicroscope. (B) Statistical scoring of rejection severity. (C) H & E staining showing pathological changes in corneal tissue at 4 weeks post-transplantation (200 × magnification).

Subsequently, metabolomics sequencing was conducted on serum samples from the rejection group and normal group (with six replicates in each). A total of 40 differential metabolites were screened in the rejection group, of which 25 were found to be upregulated and 15 were downregulated (Figure 2A). KEGG pathway enrichment analysis on the differential metabolites revealed that 240 signaling pathways were enriched, 14 of which were significantly different (P ≤ 0.05) (Figure 2B). Notably, two metabolic pathways were related to corneal graft rejection: amino acid biosynthesis and steroid hormone biosynthesis (Figure 2C). Elevated levels of the corresponding secondary metabolites – Hcy and 7α-hydroxypregnenolone, respectively – were found in the rejection group (Figure 2D).

Figure 2: Metabolomic sequencing reveals key metabolites involved in corneal transplant rejection (A) Clustering heatmap of differential metabolites. (B) KEGG enrichment scatter plot of differential metabolites. (C) Content levels of differential metabolites in the corneal transplant rejection group versus the normal control group.
Figure 2:

Metabolomic sequencing reveals key metabolites involved in corneal transplant rejection (A) Clustering heatmap of differential metabolites. (B) KEGG enrichment scatter plot of differential metabolites. (C) Content levels of differential metabolites in the corneal transplant rejection group versus the normal control group.

3.2 Transcriptomics sequencing identified TRAF1, a key gene associated with Hcy-induced corneal graft rejection

Based on the results of metabolomics sequencing analysis, we used the Hcy inhibitor DZ2002 in the rejection model and found that DZ2002 significantly reduced the opacity and edema of the graft (Figure 3A). There was a significant reduction in the RI scores at 28 days post-surgery (Figure 3B), accompanied by a reduction in inflammatory cell infiltration as seen in the HE staining (Figure 3C).

Figure 3: Screening of key genes associated with homocysteine intervention in corneal transplant rejection through transcriptomic sequencing. (A) Observation of corneal graft rejection under a stereomicroscope. (B) Statistical scoring of rejection severity. (C) H & E staining of corneal pathological changes (200 × magnification). (D) Volcano plot of differentially expressed genes. Analysis thresholds were set at |log2(Fold Change)| ≥ 1 and *p* value < 0.05. (E) GO enrichment analysis of differentially expressed genes. (F) KEGG enrichment analysis of differentially expressed genes. (G) Fold change distribution of differentially expressed genes. (H) Molecular docking model of TRAF1 with homocysteine. (I) SPR sensorgram (left) and SPR fitting curve (right).
Figure 3:

Screening of key genes associated with homocysteine intervention in corneal transplant rejection through transcriptomic sequencing. (A) Observation of corneal graft rejection under a stereomicroscope. (B) Statistical scoring of rejection severity. (C) H & E staining of corneal pathological changes (200 × magnification). (D) Volcano plot of differentially expressed genes. Analysis thresholds were set at |log2(Fold Change)| ≥ 1 and *p* value < 0.05. (E) GO enrichment analysis of differentially expressed genes. (F) KEGG enrichment analysis of differentially expressed genes. (G) Fold change distribution of differentially expressed genes. (H) Molecular docking model of TRAF1 with homocysteine. (I) SPR sensorgram (left) and SPR fitting curve (right).

To further analyze the mechanism of action, transcriptomics sequencing was performed on both the model group and the intervention group. A total of 967 differentially expressed genes were found in the model group, of which 293 were upregulated and 647 were downregulated (Figure 3D). GO and KEGG enrichment analyses were performed on these differentially expressed genes to identify the pathways related to immune rejection. Among the 292 pathways obtained, the PI3K/AKT pathway had the most significant difference (Figure 3E and F). Genes exhibiting a reverse change trend within these 48 pathways were scrutinized, leading to the identification of the following six key genes: IGF2, INSR, CSF1R, TRAF1, EPHA2, and KITLG (Figure 3G). After reviewing the literature, TRAF1 was finally selected for further investigation. Molecular docking revealed a binding site between Hcy and TRAF1 (Figure 3H).

To further confirm the interaction between TRAF1 and L-homocysteine, a surface plasmon resonance (SPR) assay was performed. The results are shown in the figure below. A series of L-homocysteine analyte concentrations (0 μM, 3.9 μM, 7.8 μM, 15.6 μM, 31.2 μM, 62.5 μM, 125 μM, 250 μM, and 500 μM) were applied to observe changes in the signal response. The sensorgram indicated that homocysteine stably binds to TRAF1. As the concentration decreased, the peak response also gradually decreased. The concentration–response fitting curve demonstrated that the signal response increased with rising analyte concentration, indicating enhanced binding affinity (Figure 3I), which suggests a dose-dependent binding relationship between homocysteine and TRAF1. The affinity constant (KD) calculated from the fitting curve was 2.46 × 10−4 M, confirming a measurable binding ability between homocysteine and TRAF1.

3.3 Hcy was found to activate the PI3K/AKT pathway through TRAF1 to promote Th17 differentiation and corneal graft rejection

3.3.1 Identifying the possible inflammation-regulatory substances in the aqueous humor of rats treated with DZ2002 for corneal graft rejection

To clarify whether Hcy activates the PI3K/AKT pathway through TRAF1 to promote Th17 differentiation and corneal graft rejection, we first used a multiplex kit to detect the expression of inflammatory factors in the aqueous humor of rats treated with DZ2002 for corneal graft rejection and screened the following four factors related to TRAF1: IL1A, C–C motif chemokine ligand 5 (CCL5), TNFRSF1A, and TNFRSF1B (Figure 4A).

Figure 4: Screening of potential inflammatory mediators in aqueous humor following DZ2002 intervention in corneal rejection. (A) Expression profile of inflammatory factors detected by multiplex assay. (B) GO enrichment analysis of differentially expressed inflammatory factors. (C) KEGG enrichment analysis of differentially expressed inflammatory factors. (D) Protein-protein interaction (PPI) network between TRAF1 and relevant inflammatory factors. (E) RT-PCR validation of significantly altered inflammatory factors in corneal tissue, using β-actin as internal reference. (F) ELISA quantification of serum inflammatory factors IL-1β, IL-6, and TGF-β.
Figure 4:

Screening of potential inflammatory mediators in aqueous humor following DZ2002 intervention in corneal rejection. (A) Expression profile of inflammatory factors detected by multiplex assay. (B) GO enrichment analysis of differentially expressed inflammatory factors. (C) KEGG enrichment analysis of differentially expressed inflammatory factors. (D) Protein-protein interaction (PPI) network between TRAF1 and relevant inflammatory factors. (E) RT-PCR validation of significantly altered inflammatory factors in corneal tissue, using β-actin as internal reference. (F) ELISA quantification of serum inflammatory factors IL-1β, IL-6, and TGF-β.

Subsequent GO and KEGG enrichment analyses of the differential inflammatory factors yielded a total of 20 significant signals, six of which were disease-related (Figure 4B and C). The Protein-Protein Interaction (PPI) network analysis demonstrated interactions between TRAF1 and these four factors (Figure 4D).

The expression levels of the four factors in corneal tissues were further detected using RT-PCR. This revealed higher expressions of IL1A, CCL5, and TNFRSF1A in the model group than in the control group. Conversely, the expressions of IL1A, CCL5, and TNFRSF1A were lower in the intervention group than in the model group. Additionally, the expressions of IL1A, CCL5, and TNFRSF1A in the intervention + OE-TRAF1 group were higher than those in the intervention group, whereas notably, TNFRSF1B expression did not differ significantly across groups (Figure 4E).

Furthermore, the levels of inflammatory factors IL-1β, IL-6, and TGF-β were detected using ELISA. There was a significant increase in the levels of IL-1β, IL-6, and TGF-β in the model group starting on day 7 post-surgery. Conversely, in the intervention group, the levels of IL-1β, IL-6, and TGF-β decreased after day 14, and in the intervention + OE-TRAF1 group, the levels of IL-1β, IL-6, and TGF-β increased again after day 14 (Figure 4F).

3.3.2 Hcy activated the PI3K/AKT pathway through TRAF1 to promote corneal graft rejection

The effect of simultaneous intervention with DZ2002 and TRAF1 on corneal graft rejection was subsequently observed.

Compared with the DZ2002 group, the corneal graft in the DZ2002 + OE-TRAF1 group had increased opacity and edema, as observed under a stereomicroscope (Figure 5A). The RI scores were significantly elevated at four weeks (Figure 5B), and HE results showed that the inflammatory response was aggravated in the DZ2002 + OE-TRAF1 group (Figure 5C). WB analysis verified that the protein expressions of TRAF1, p-PI3K/PI3K, and p-AKT/AKT were significantly increased in the model group, whereas the protein expression levels of TRAF1, p-PI3K/PI3K, and p-AKT/AKT decreased after DZ2002 intervention. However, the expressions of TRAF1, p-PI3K/PI3K, and p-AKT/AKT were reversed following simultaneous intervention with DZ2002 and TRAF1 (Figure 5D). The effects of simultaneous intervention with DZ2002 and TRAF1 on angiogenesis and apoptosis levels were also additionally examined. Compared with the DZ2002 group, the VEGF expression levels and apoptosis rates were significantly higher in the DZ2002 + OE-TRAF1 group (Figure 5E and F). These results suggest that Hcy induced the activation of the PI3K/AKT pathway by promoting the expression of TRAF1, thereby leading to corneal graft rejection.

Figure 5: Effects of combined DZ2002 and TRAF1 intervention on corneal allograft rejection (A) Stereomicroscopic evaluation of corneal graft rejection. (B) Statistical scoring of rejection severity. (C) H & E staining demonstrating corneal pathological changes (200 × magnification). (D) Western blot analysis of TRAF1, PI3K, and AKT protein expression levels, with β-actin as the loading control. (E) Immunohistochemical (IHC) detection of VEGF positive rate (200 × magnification). (F) TUNEL staining for assessment of apoptotic levels (200 × magnification).
Figure 5:

Effects of combined DZ2002 and TRAF1 intervention on corneal allograft rejection (A) Stereomicroscopic evaluation of corneal graft rejection. (B) Statistical scoring of rejection severity. (C) H & E staining demonstrating corneal pathological changes (200 × magnification). (D) Western blot analysis of TRAF1, PI3K, and AKT protein expression levels, with β-actin as the loading control. (E) Immunohistochemical (IHC) detection of VEGF positive rate (200 × magnification). (F) TUNEL staining for assessment of apoptotic levels (200 × magnification).

3.3.3 In vitro cell experiments confirmed that Hcy activates the PI3K/AKT pathway through TRAF1 to promote Th17 differentiation

To explore the mechanism of corneal graft rejection, in vitro cell experiments were conducted to validate that Hcy activates the PI3K/AKT pathway through TRAF1. Given the pivotal role of CD4+ T cell polarization in immune imbalance, the effect of human corneal epithelial cells (HCEC) on CD4+ T cell polarization post-DZ2002 and TRAF1 intervention was examined via flow cytometry. The results showed that, in the HCEC + OE-TRAF1 group compared with the HCEC + OE-NC group, Th17 cells increased whereas Treg decreased. Also, compared with the HCEC + OE-NC + DZ2002 group, Th17 cells increased and Treg decreased in the HCEC + OE-TRAF1 + DZ2002 group, indicating that the increased expression level of TRAF1 could promote the differentiation of CD4+ T cells into Th17. In the HCEC + OE-TRAF1 + DZ2002 group, compared with the HCEC + OE-TRAF1 group, Th17 cells decreased and Treg increased, suggesting that DZ2002 could inhibit the regulation of CD4 cell polarization by TRAF1 (Figure 6A and B).

Figure 6: Mechanistic insights into Hcy-induced corneal rejection via TRAF1-mediated activation of the PI3K/AKT pathway (A) Flow cytometric analysis of TH17 cell proportion. (B) Flow cytometric analysis of Treg cell proportion. (C) Western blot analysis of TRAF1, PI3K, and AKT protein expression, using β-actin as the loading control.
Figure 6:

Mechanistic insights into Hcy-induced corneal rejection via TRAF1-mediated activation of the PI3K/AKT pathway (A) Flow cytometric analysis of TH17 cell proportion. (B) Flow cytometric analysis of Treg cell proportion. (C) Western blot analysis of TRAF1, PI3K, and AKT protein expression, using β-actin as the loading control.

WB was used to assess the protein expression levels of PI3K and AKT in HCEC cells co-cultured with CD4+ T cells. The results showed that, compared with the HCEC + OE-NC group, the expressions of TRAF1, p-PI3K/PI3K, and p-AKT/AKT were significantly increased in the HCEC + OE-TRAF1 group. Decreased levels of TRAF1, p-PI3K/PI3K, and p-AKT/AKT were observed in the HCEC + OE-TRAF1 + DZ2002 group when compared with the HCEC + OE-NC + DZ2002 group (Figure 6C). These results suggest that Hcy activates the PI3K/AKT pathway by upregulating TRAF1, thereby contributing to corneal graft rejection and increased Th17 differentiation.

4 Discussion

The treatment of corneal graft rejection aims to restore the normal metabolic processes of corneal tissue and preserve corneal graft transparency. A primary focus of research in this area revolves around reducing the destructive attack of CD4+ T cells on the graft [8], [9], [10]. Strategies such as injecting the alpha-melanocyte stimulating hormone (α-MSH), vasoactive intestinal peptide, transforming growth factor β2 (TGF-β2), and thromboxane-1 (TSP-1) into the anterior chamber have been explored to inhibit lymphatic production and regulate T cell activity through different signaling pathways, thereby maintaining the immune privilege of the anterior chamber [11]. Although these approaches have shown promising results, they are still associated with challenges such as high cost, technical complexity, and limited scalability.

When a corneal graft rejection occurs, the patient’s immune system identifies the transplanted corneal tissue as a foreign substance and initiates inflammatory responses to eliminate it. Th1, Th2, and Th17 (CD4+ CD25+ RORγt+) cells play crucial roles in this inflammatory cascade, releasing a variety of cytokines and chemical factors such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-17 (IL-17), TGF-β, and interleukin-21 (IL-21). These factors further regulate the immune response and inflammation, accompanied by massive corneal neovascularization and increased VEGF factor expression [12], [13], [14], [15]. Consistent with previous studies, in our study, HE staining of the corneal tissue with graft rejection showed significant infiltration of inflammatory cells. Also, levels of IL-1β, IL-6, and TGF-β were significantly elevated since day 7 post-surgery, while VEGF expression and corneal apoptosis were significantly increased at four weeks post-surgery.

The use of metabolomics in the investigation of corneal diseases represents a novel approach that offers new perspectives and methodologies for uncovering the pathogenesis, diagnosis, and treatment of these conditions. Corneal epithelial cells, stromal cells, and endothelial cells collectively engage in protein synthesis and metabolic processes crucial for maintaining the normal physiological functioning of the cornea. Any pathological state that interferes with corneal protein metabolism may lead to corneal opacity and vision loss [16].

4.1 The crucial role of Hcy in corneal graft rejection

In this study, metabolomics analysis performed on the serum of rats with rejection revealed two metabolic pathways related to corneal graft rejection: amino acid and steroid hormone biosynthesis. The corresponding secondary metabolites were Hcy and 7α-hydroxypregnenolone, respectively. However, no direct association has been reported between 7α-OH-DHEA and corneal diseases. Hcy is a sulfur-containing amino acid that is involved in vivo protein metabolism processes. It can be metabolized to methionine or reconverted to methionine through the action of vitamins B6, B12, and folic acid.

Abnormally elevated Hcy levels are associated with increased risks of a variety of diseases, including cardiovascular, neurological, and ocular diseases [17]. Although studies have linked Hcy to conditions such as keratoconus, dry eye, and corneal ulcers, there are relatively few studies in the field of corneal transplantation. However, there is emerging evidence suggesting that Hcy may influence immune cell function through multiple mechanisms, such as modulating cellular oxidative stress levels, and consequently indirectly affecting the activation and differentiation of T cells [18]. Notably, Wolos et al. [19] demonstrated that S-adenosylhomocysteine hydrolase inhibitors can affect immune rejection in vitro and in vivo. The results of this study underscore the role of Hcy as a key metabolite potentially implicated in promoting the occurrence of rejection through some mechanism, thus providing new clues for further exploration of the molecular mechanisms involved in corneal graft rejection.

4.2 Activation of TRAF1 and PI3K/AKT pathways

Post-corneal transplantation, rats was administered DZ2002 via eye drops at a frequency of four times per day. After four weeks of continuous use, transcriptomics sequencing was used to identify the key genes associated with Hcy influencing corneal graft rejection and found that the expression of TRAF1 was significantly reduced after the DZ2002 intervention. GO and KEGG enrichment analyses of the differentially expressed genes identified the PI3K/AKT pathway to be the most significant among the pathways related to immune rejection. Molecular docking analysis showed that there was a binding site between Hcy and TRAF1. We have successfully validated, for the first time, the specific binding between Hcy and TRAF1 protein using surface plasmon resonance (SPR). Therefore, we further speculated that DZ2002 inhibited the binding of Hcy to TRAF1 during corneal graft rejection.

The exact mechanism of TRAF1 in CD4+ T cell differentiation and function is still being studied. It is thought to be an important molecule in regulating CD4+ T cell-mediated immune responses and possible immunopathological processes [20], 21]. A variety of intracellular signaling pathways regulate Th17 cell differentiation, including the complex transcription factor network involving PI3K, AKT, and mTOR [22], [23], [24]. However, the increased expression of these inflammatory indicators after the addition of the TRAF1AVV virus in the DZ2002 group indicates that DZ2002 mainly reduced the inflammatory response by downregulating the expression of TRAF1.

Immune cells, notably CD4+ T cells, migrate to corneal grafts – a process that constitutes a fundamental step in the cascade leading to immune rejection. Chemokines not only attract the migration and localization of CD4+ T cells but also affect their differentiation and exert partial immune regulatory effects, thus playing a key role in the immune response after corneal transplantation [25]. In the current study, we employed a multiplex kit to detect the expression levels of inflammatory factors in the aqueous humor of rats treated with DZ2002 for graft rejection, and this identified four inflammatory cytokines, namely, IL1A, CCL5, TNFRSF1A, and TNFRSF1B, associated with TRAF1. These findings offer important clues for further research on the role of Hcy in corneal graft rejection. In this study, GO and KEGG enrichment analyses of differential inflammatory factors were performed, and RT-PCR results verified that IL1A, CCL5, and TNFRSF1A were significantly reduced in the DZ2002 group. Meanwhile, ELISA revealed a reduction in all the inflammatory factors, i.e., IL-1β, IL-6, and TGF-β, underscoring the significant impact of DZ2002 on inhibiting the inflammatory response of corneal transplantation.

The inflammatory regulatory role of TRAF1 is cell type- and pathology-dependent. It primarily exerts anti-inflammatory effects in cardiomyocytes, macrophages, and T cells, while exhibiting pro-inflammatory effects in endothelial cells and certain tumor models. The underlying mechanisms involve NF-κB and MAPK signaling pathways, as well as interactions with cIAP2 [26], 27]. This dual functionality may be attributed to differences in the adapter molecules that TRAF1 interacts with across distinct cell types [28].

In this study, overexpression of TRAF1 on the basis of DZ2002 intervention significantly upregulated the expression of IL-1A, CCL5, and TNFRSF1A in corneal tissue. Furthermore, levels of the inflammatory factors IL-1β and IL-6, as well as TGF-β, were markedly elevated in aqueous humor. TGF-β is a multifunctional cytokine produced and secreted by a wide range of cell types, demonstrating broad cellular origins and functional diversity. It is primarily secreted by M2 macrophages, dendritic cells (DCs), T cells, as well as various epithelial and vascular cells [29], [30], [31]. The results indicate that TRAF1 promotes the expression of inflammatory factors in corneal rejection and activates the secretory functions of both immune cells and corneal epithelial cells.

Studies have shown that the damaged corneal tissues with inflammation after corneal transplantation would express a series of chemokines, including CCL2, CCL3, CCL4, CCL5, and CXCL10 [25], 32], 33]. In this study, our focus was on the examination and validation of CCL5, a TRAF1-related chemokine. Our findings suggest that TRAF1 has a potential role in promoting rejection by increasing the expression of CCL5. CCL5, also known as Regulated upon Activation, Normal T Cell Expressed and Secreted (RANTES), is a small pro-inflammatory cytokine belonging to the CC chemokine family. It is secreted by various cells, including T cells, macrophages, and fibroblasts, and plays an important role in immune responses and inflammatory processes [34]. Mechanistically, CCL5 exerts its effects by binding to specific chemokine receptors such as CCR1, CCR3, and CCR5. It has a strong chemotaxis effect on various immune cells, including monocytes, T cells, eosinophils, and so on, thereby facilitating their migration to sites of inflammation or infection.

In the present experiment, it was observed that the phosphorylation level of PI3K/AKT and CCL5 was decreased in the DZ2002 group, suggesting that the activation of the PI3K/AKT pathway was inhibited. Consequently, VEGF expression and corneal apoptosis rates were reduced. Additionally, in the co-culture of corneal epithelial cells and CD4+ T cells, we confirmed that DZ2002 could inhibit the differentiation of CD4+ T cells into Th17 by downregulating the expression of TRAF1.

4.3 Potential therapeutic effects of DZ2002

The effects of Hcy on immune regulation may be attributed to its involvement in several of the following underlying mechanisms: Elevated Hcy levels can increase oxidative stress in the body, activating immune cells such as macrophages and lymphocytes. This may lead to an exacerbation of the inflammatory response, thereby resulting in cell damage. This oxidative stress can increase the production of inflammatory mediators, including cytokines and chemokines [35], 36], which regulate immune cell activity [37], 38] and indirectly affect the immune response by influencing vascular endothelial cell function [39], [40], [41], [42]. Additionally, Hcy has been implicated in the development of autoimmune diseases [43], 44]. The in vivo metabolism of Hcy involves methylation processes and aberrations in these may affect gene expression patterns, including those that regulate the immune system, thereby affecting the function and development of immune cells [45].

DZ2002 has been shown to modulate the STAT3-PI3K-Akt-NF-κB signaling pathway, resulting in reduced expression of genes associated with angiogenesis and decreased levels of inflammation in the cornea and conjunctiva. This modulation helps alleviate dry eye symptoms [46]. Furthermore, in this present study, DZ2002 effectively inhibited corneal graft rejection, suggesting that DZ2002 may be a potential drug for the treatment of corneal graft rejection.

Although some progress has been made in this study, there are still many issues that warrant further exploration. For example, uncovering the specific mechanisms of action of the Hcy, TRAF1, and PI3K/AKT pathways in corneal graft rejection requires a more in-depth study. Additionally, its application potential and safety profile in clinical practice need further investigations.

One limitation of this study should be noted. The absence of a syngeneic transplantation control group (e.g., SD rats to SD rats) makes it difficult to entirely rule out the potential influence of non-specific surgical trauma-induced inflammation on the early observed outcomes. Nevertheless, the progressively worsening pathological process, the specific molecular profile highly consistent with allogeneic immune responses, and the significant therapeutic effect of DZ2002 collectively indicate that the core findings of this study are primarily attributable to allogeneic immune rejection rather than merely postoperative inflammation [47]. Future studies should incorporate a syngeneic control group to establish a definitive baseline and provide critical evidence for further validating the specific role of the Hcy–TRAF1 axis in rejection responses.

5 Conclusion

In this study, employing a metabolomic approach, the metabolite Hcy and the differentially expressed gene TRAF1 were identified in the context of their role in corneal graft rejection. Experiments using a rat model of graft rejection confirmed that inhibiting Hcy led to a decrease in TRAF1 expression, lower levels of the chemokine CCL5 in the aqueous humor, suppression of the differentiation of T cells into Th17, and a subsequent reduction in the rejection index. A preliminary exploration of the DZ2002-related mechanisms involved in alleviating rejection revealed that homotype DZ2002 was effective in reducing rejection, offering promise for this to emerge as a new drug for the treatment of corneal graft rejection.

Corresponding author: Lan Li, Department of Ophthalmology, The Affiliated Calmette Hospital of Kunming Medical University, No. 1228 Beijing Road, Panlong District, Kunming 650000, China, E-mail:

Acknowledgments

We would like to acknowledge the hard and dedicated work of all the staff that implemented the intervention and evaluation components of the study.

  1. Funding information: This work was supported by Major Science and Technology Special Plan Project of Yunnan Provincial Department of Science and Technology (No.202302AA310018).

  2. Author contribution: Conception and design of the research: Lan Li, Qian Cao. Acquisition of data: Ermiao Tian, Junjun Long, Shuang Tan, Yong Li, Jie Dong. Analysis and interpretation of the data: Qian Cao, Yongqian Zhang, Ermiao Tian, Junjun Long, Yong Li. Statistical analysis: Qian Cao, Shuang Tan, Yongqian Zhang, Jie Dong. Obtaining financing: Lan Li. Writing of the manuscript: Qian Cao, Lan Li. Critical revision of the manuscript for intellectual content: Lan Li, Qian Cao. All authors read and approved the final draft.

  3. Conflict of interest: Authors state no conflict of interest.

  4. Data availability statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

1. Kacham, S, Bhure, TS, Eswaramoorthy, SD, Naik, G, Rath, SN, Parcha, SR, et al.. Human umbilical cord-derived mesenchymal stem cells promote corneal epithelial repair in vitro. Cells 2021;10:1254. https://doi.org/10.3390/cells10051254.Search in Google Scholar PubMed PubMed Central

2. Bourne, RRA, Flaxman, SR, Braithwaite, T, Cicinelli, MV, Das, A, Jonas, JB, et al.. Magnitude, temporal trends, and projections of the global prevalence of blindness and distance and near vision impairment: a systematic review and meta-analysis. Lancet Glob Health 2017;5:e888–97. https://doi.org/10.1016/s2214-109x(17)30293-0.Search in Google Scholar PubMed

3. Pascolini, D, Mariotti, SP. Global estimates of visual impairment:2010. Br J Ophthalmol 2012;96:614–8. https://doi.org/10.1136/bjophthalmol-2011-300539.Search in Google Scholar PubMed

4. Guo, YX, Hong, J. [Graft virus infection following corneal transplantation]. [Zhonghua yan ke za zhi] Chin J Ophthalmol 2019;55:713–16. https://doi.org/10.3760/cma.j.issn.0412-4081.2019.09.018.Search in Google Scholar PubMed

5. Tan, DTH, Dart, JKG, Holland, EJ, Kinoshita, S. Corneal transplantation. Lancet 2012;379:1749–61. https://doi.org/10.1016/S0140-6736(12)60437-1.Search in Google Scholar PubMed

6. Amouzegar, A, Chauhan, SK, Dana, R. Alloimmunity and tolerance in corneal transplantation. J Immunol 2016;196:3983–91. https://doi.org/10.4049/jimmunol.1600251.Search in Google Scholar PubMed PubMed Central

7. Holland, EJ, Chan, CC, Wetzig, RP, Palestine, AG, Nussenblatt, RB. Clinical and immunohistologic studies of corneal rejection in the rat penetrating keratoplasty model. Cornea 1991;10:374–80. https://doi.org/10.1097/00003226-199109000-00003.Search in Google Scholar PubMed

8. Pastak, M, Kleff, V, Saban, DR, Czugala, M, Steuhl, KP, Ergün, S, et al.. Gene therapy for modulation of T-Cell-mediated immune response provoked by corneal transplantation. Hum Gene Ther 2018;29:467–79. https://doi.org/10.1089/hum.2017.044.Search in Google Scholar PubMed PubMed Central

9. Koudouna, E, Okumura, N, Okazaki, Y, Nakano, S, Inoue, R, Fullwood, NJ, et al.. Immune cells on the corneal endothelium of an allogeneic corneal transplantation rabbit model. Investig Ophthalmol Vis Sci 2017;58:242–51. https://doi.org/10.1167/iovs.16-20019.Search in Google Scholar PubMed

10. Hegde, S, Beauregard, C, Mayhew, E, Niederkorn, JY. CD4(+) T-cell-mediated mechanisms of corneal allograft rejection: role ofFas-induced apoptosis. Transplantation 2005;79:23–31. https://doi.org/10.1097/01.tp.0000147196.79546.69.Search in Google Scholar PubMed

11. Hori, J, Yamaguchi, T, Keino, H, Hamrah, P, Maruyama, K. Immune privilege in corneal transplantation. Prog Retin Eye Res 2019;72:100758. https://doi.org/10.1016/j.preteyeres.2019.04.002.Search in Google Scholar PubMed

12. Sun, J, Wang, T, Bian, J, Shi, W, Ruan, Q. Immune tolerance induced in the anterior chamber ameliorates corneal transplant rejection. Clin Immunol 2023;257:109797. https://doi.org/10.1016/j.clim.2023.109797.Search in Google Scholar PubMed

13. Wang, X, Wang, W, Xu, J, Wu, S, Le, Q. All-trans retinoid acid promotes allogeneic corneal graft survival in mice by regulating Treg-Th17 balance in the presence of TGF-beta. BMC Immunol 2015;16:17. https://doi.org/10.1186/s12865-015-0082-3.Search in Google Scholar PubMed PubMed Central

14. Cunnusamy, K, Chen, PW, Niederkorn, JY. IL-1β7A-dependent CD4+CD25+ regulatory T cells promote immune privilege of corneal allografts. J Immunol 2011;186:6737–45. https://doi.org/10.4049/jimmunol.1100101.Search in Google Scholar PubMed PubMed Central

15. Jiang, L, Liu, T, Xie, L, Ouyang, C, Ji, J, Huang, T. AICAR prolongs corneal allograft survival via the AMPK-mTOR signaling pathway in mice. Biomed Pharmacother 2019;113:108558. https://doi.org/10.1016/j.biopha.2019.01.019.Search in Google Scholar PubMed

16. Teo, AWJ, Zhang, J, Zhou, L, Liu, YC. Metabolomics in corneal diseases: a narrative review from clinical aspects. Metabolites 2023;13:380. https://doi.org/10.3390/metabo13030380.Search in Google Scholar PubMed PubMed Central

17. Smith, AD, Refsum, H. Homocysteine – from disease biomarker to disease prevention. J Intern Med 2021;290:826–54. https://doi.org/10.1111/joim.13279.Search in Google Scholar PubMed

18. Gao, X, Li, J, Chen, M. Effect of homocysteine on the differentiation of CD4+ T cells into Th17 cells. Dig Dis Sci 2018;63:3339–47. https://doi.org/10.1007/s10620-018-5177-2.Search in Google Scholar PubMed

19. Wolos, JA, Frondorf, KA, Babcock, GF, Stripp, SA, Bowlin, TL. Immunomodulation by an inhibitor of S-adenosyl-L-homocysteine hydrolase: inhibition of in vitro and in vivo allogeneic responses. Cell Immunol 1993;149:402–8. https://doi.org/10.1006/cimm.1993.1165.Search in Google Scholar PubMed

20. Park, H. Structure of TRAF family: current understanding of receptor recognition. Front Immunol. 2018:9:1999, 1999. https://doi.org/10.3389/fimmu.2018.01999.Search in Google Scholar PubMed PubMed Central

21. McPherson, AJ, Snell, LM, Mak, TW, Watts, TH. Opposing roles for TRAF1 in the alternative versus classical NF-κB pathway in T cells. J Biol Chem 2012;287:23010–9. https://doi.org/10.1074/jbc.M112.350538.Search in Google Scholar PubMed PubMed Central

22. Jiang, Y, Wang, X, Dong, C. Molecular mechanisms of T helper 17 cell differentiation: emerging roles for transcription cofactors. Adv Immunol 2019;144:121–53. https://doi.org/10.1016/bs.ai.2019.09.003.Search in Google Scholar PubMed

23. Gao, L, Dong, Y, Lin, R, Meng, Y, Wu, F, Jia, L. The imbalance of Treg/Th17 cells induced by perinatal bisphenol A exposure is associated with activation of the PI3K/Akt/mTOR signaling pathway in male offspring mice. Food Chem Toxicol 2020;137:111177. https://doi.org/10.1016/j.fct.2020.111177.Search in Google Scholar PubMed

24. Chung, Y, Chang, SH, Martinez, GJ, Yang, XO, Nurieva, R, Kang, SH, et al.. Critical regulation of early Th17 cell differentiation by interleukin-1 signaling. Immunity 2009;30:576–87. https://doi.org/10.1016/j.immuni.2009.02.007.Search in Google Scholar PubMed PubMed Central

25. Lai, Q, Hu, L, Zhang, W, Jiang, Z, Zeng, C, Hu, J. Single-cell RNA sequencing highlights the regulatory role of T cell marker genes Ctla4, Ccl5 and Tcf7 in corneal allograft rejection of mouse model. Int Immunopharmacol 2023;117:109911. https://doi.org/10.1016/j.intimp.2023.109911.Search in Google Scholar PubMed

26. Mirzaesmaeili, A, Abdul-Sater, AA. TRAF1 deficiency in macrophages drives exacerbated joint inflammation in rheumatoid arthritis. Biomolecules 2024;14:864. https://doi.org/10.3390/biom14070864.Search in Google Scholar PubMed PubMed Central

27. Tang, Y, Aleithan, F, Madahar, SS, Mirzaesmaeili, A, Saran, S, Tang, J, et al.. Selective disruption of Traf1/cIAP2 interaction attenuates inflammatory responses and rheumatoid arthritis. J Autoimmun 2025;152:103377. https://doi.org/10.1016/j.jaut.2025.103377.Search in Google Scholar PubMed

28. Edilova, MI, Abdul-Sater, AA, Watts, TH. TRAF1 signaling in human health and disease. Front Immunol 2018;9:2969. https://doi.org/10.3389/fimmu.2018.02969.Search in Google Scholar PubMed PubMed Central

29. Krneta, T, Gillgrass, A, Poznanski, S, Chew, M, Lee, AJ, Kolb, M, et al.. M2-polarized and tumor-associated macrophages alter NK cell phenotype and function in a contact-dependent manner. J Leukoc Biol 2017;101:285–95. https://doi.org/10.1189/jlb.3A1215-552R.Search in Google Scholar PubMed

30. Ihara, S, Hirata, Y, Koike, K. TGF-β in inflammatory bowel disease: a key regulator of immune cells, epithelium, and the intestinal microbiota. J Gastroenterol 2017;52:777–87. https://doi.org/10.1007/s00535-017-1350-1.Search in Google Scholar PubMed

31. Frangogiannis, N. Transforming growth factor-β in tissue fibrosis. J Exp Med 2020;217:e20190103. https://doi.org/10.1084/jem.20190103.Search in Google Scholar PubMed PubMed Central

32. King, WJ, Comer, RM, Hudde, T, Larkin, DF, George, AJ. Cytokine and chemokine expression kinetics after corneal transplantation. Transplantation 2000;70:1225–33. https://doi.org/10.1097/00007890-200010270-00017.Search in Google Scholar PubMed

33. Yamagami, S, Hamrah, P, Zhang, Q, Liu, Y, Huq, S, Dana, MR. Early ocular chemokine gene expression and leukocyte infiltration after high-risk corneal transplantation. Mol Vis 2005;11:632–40.Search in Google Scholar

34. Barczak, K, Droździk, A, Bosiacki, M, Łagocka, R, Cenariu, D, Uriciuc, WA, et al.. CCL5’s role in periodontal disease: a narrative review. Int J Mol Sci 2023;24:17332. https://doi.org/10.3390/ijms242417332.Search in Google Scholar PubMed PubMed Central

35. Ye, M, Li, H, Luo, H, Zhou, Y, Luo, W, Lin, Z. Potential antioxidative activity of homocysteine in erythrocytes under oxidative stress. Antioxidants (Basel) 2023;12:202. https://doi.org/10.3390/antiox12010202.Search in Google Scholar PubMed PubMed Central

36. Li, J, Zhang, Y, Zhang, Y, Lü, S, Miao, Y, Yang, J, et al.. GSNOR modulates hyperhomocysteinemia-induced T cell activation and atherosclerosis by switching Akt S-nitrosylation to phosphorylation. Redox Biol 2018;17:386–99. https://doi.org/10.1016/j.redox.2018.04.021.Search in Google Scholar PubMed PubMed Central

37. Faghfouri, AH, Zarezadeh, M, Tavakoli-Rouzbehani, OM, Radkhah, N, Faghfuri, E, Kord-Varkaneh, H, et al.. The effects of N-acetylcysteine on inflammatory and oxidative stress biomarkers: a systematic review and meta-analysis of controlled clinical trials. Eur J Pharmacol 2020;884:173368. https://doi.org/10.1016/j.ejphar.2020.173368.Search in Google Scholar PubMed

38. Borowska, M, Winiarska, H, Dworacka, M, Wesołowska, A, Dworacki, G, Mikołajczak, PŁ. The effect of homocysteine on the secretion of Il-1β, Il-6, Il-10, Il-12 and RANTES by peripheral blood mononuclear cells-an in vitro study. Molecules 2021;26:6671. https://doi.org/10.3390/molecules26216671.Search in Google Scholar PubMed PubMed Central

39. Jiang, X, Yang, F, Brailoiu, E, Jakubowski, H, Dun, NJ, Schafer, AI, et al.. Differential regulation of homocysteine transport in vascular endothelial and smooth muscle cells. Arterioscler Thromb Vasc Biol 2007;27:1976–83. https://doi.org/10.1161/ATVBAHA.107.148544.Search in Google Scholar PubMed PubMed Central

40. Leng, Y, Chen, R, Chen, R, He, S, Shi, X, Zhou, X, et al.. HMGB1 mediates homocysteine-induced endothelial cells pyroptosis via cathepsin V-dependent pathway. Biochem Biophys Res Commun 2020;532:640–6. https://doi.org/10.1016/j.bbrc.2020.08.091.Search in Google Scholar PubMed

41. Zhang, D, Hong, X, Wang, J, Jiang, Y, Zhang, Y, Chen, J, et al.. Estradiol-17β inhibits homocysteine mediated damage by promoting H2S production via upregulating CBS and CSE expression in human umbilical vein endothelial cells. J Cell Biochem 2021;122:915–25. https://doi.org/10.1002/jcb.29527.Search in Google Scholar PubMed

42. Chatterjee, B, Fatima, F, Seth, S, Roy, SS. Moderate elevation of homocysteine induces endothelial dysfunction through adaptive UPR activation and metabolic rewiring. Cells 2024;13:214. https://doi.org/10.3390/cells13030214.Search in Google Scholar PubMed PubMed Central

43. Zhang, YM, Zhou, XJ, Shi, SF, Liu, LJ, Lyu, JC, Zhang, H. Homocysteine and IgA nephropathy: observational and Mendelian randomization analyses. Chin Med J (Engl) 2020;133:277–84. https://doi.org/10.1097/CM9.0000000000000613.Search in Google Scholar PubMed PubMed Central

44. Álvarez-Cienfuegos, A, Cantero-Nieto, L, García-Gómez, JA, Sabio, JM, González-Gay, MA, Ortego-Centeno, N. Association between homocysteine levels and arterial stiffness in women with systemic sclerosis. Clin Exp Rheumatol 2020;38:92–7.Search in Google Scholar

45. Mohammad, G, Kowluru, RA. Homocysteine disrupts balance between MMP-9 and its tissue inhibitor in diabetic retinopathy: the role of DNA methylation. Int J Mol Sci 2020;21:1771. https://doi.org/10.3390/ijms21051771.Search in Google Scholar PubMed PubMed Central

46. Wu, CM, Mao, JW, Zhu, JZ, Xie, CC, Yao, JY, Yang, XQ, et al.. DZ2002 alleviates corneal angiogenesis and inflammation in rodent models of dry eye disease via regulating STAT3-PI3K-Akt-NF-κB pathway. Acta Pharmacol Sin 2023;45:166–79. https://doi.org/10.1038/s41401-023-01146-y.Search in Google Scholar PubMed PubMed Central

47. Tian, H, Lin, S, Wu, J, Ma, M, Yu, J, Zeng, Y, et al.. Kaempferol alleviates corneal transplantation rejection by inhibiting NLRP3 inflammasome activation and macrophage M1 polarization via promoting autophagy. Exp Eye Res 2021;208:108627. https://doi.org/10.1016/j.exer.2021.108627.Search in Google Scholar PubMed

Received: 2025-07-30
Accepted: 2025-10-07
Published Online: 2026-01-01

© 2025 the author(s), published by De Gruyter, Berlin/Boston

This work is licensed under the Creative Commons Attribution 4.0 International License.

Articles in the same Issue

  1. Safety assessment and modulation of hepatic CYP3A4 and UGT enzymes by Glycyrrhiza glabra aqueous extract in female Sprague–Dawley rats
  2. Adult-onset Still’s disease with hemophagocytic lymphohistiocytosis and minimal change disease
  3. Role of DZ2002 in reducing corneal graft rejection in rats by influencing Th17 activation via inhibition of the PI3K/AKT pathway and downregulation of TRAF1
  4. Biomedical Sciences
  5. Mechanism of triptolide regulating proliferation and apoptosis of hepatoma cells by inhibiting JAK/STAT pathway
  6. Maslinic acid improves mitochondrial function and inhibits oxidative stress and autophagy in human gastric smooth muscle cells
  7. Comparative analysis of inflammatory biomarkers for the diagnosis of neonatal sepsis: IL-6, IL-8, SAA, CRP, and PCT
  8. Post-pandemic insights on COVID-19 and premature ovarian insufficiency
  9. Proteome differences of dental stem cells between permanent and deciduous teeth by data-independent acquisition proteomics
  10. Optimizing a modified cetyltrimethylammonium bromide protocol for fungal DNA extraction: Insights from multilocus gene amplification
  11. Preliminary analysis of the role of small hepatitis B surface proteins mutations in the pathogenesis of occult hepatitis B infection via the endoplasmic reticulum stress-induced UPR-ERAD pathway
  12. Efficacy of alginate-coated gold nanoparticles against antibiotics-resistant Staphylococcus and Streptococcus pathogens of acne origins
  13. Battling COVID-19 leveraging nanobiotechnology: Gold and silver nanoparticle–B-escin conjugates as SARS-CoV-2 inhibitors
  14. Neurodegenerative diseases and neuroinflammation-induced apoptosis
  15. Impact of fracture fixation surgery on cognitive function and the gut microbiota in mice with a history of stroke
  16. COLEC10: A potential tumor suppressor and prognostic biomarker in hepatocellular carcinoma through modulation of EMT and PI3K-AKT pathways
  17. High-temperature requirement serine protease A2 inhibitor UCF-101 ameliorates damaged neurons in traumatic brain-injured rats by the AMPK/NF-κB pathway
  18. SIK1 inhibits IL-1β-stimulated cartilage apoptosis and inflammation in vitro through the CRTC2/CREB1 signaling
  19. Rutin–chitooligosaccharide complex: Comprehensive evaluation of its anti-inflammatory and analgesic properties in vitro and in vivo
  20. Knockdown of Aurora kinase B alleviates high glucose-triggered trophoblast cells damage and inflammation during gestational diabetes
  21. Calcium-sensing receptors promoted Homer1 expression and osteogenic differentiation in bone marrow mesenchymal stem cells
  22. ABI3BP can inhibit the proliferation, invasion, and epithelial–mesenchymal transition of non-small-cell lung cancer cells
  23. Changes in blood glucose and metabolism in hyperuricemia mice
  24. Rapid detection of the GJB2 c.235delC mutation based on CRISPR-Cas13a combined with lateral flow dipstick
  25. IL-11 promotes Ang II-induced autophagy inhibition and mitochondrial dysfunction in atrial fibroblasts
  26. Short-chain fatty acid attenuates intestinal inflammation by regulation of gut microbial composition in antibiotic-associated diarrhea
  27. Application of metagenomic next-generation sequencing in the diagnosis of pathogens in patients with diabetes complicated by community-acquired pneumonia
  28. NAT10 promotes radiotherapy resistance in non-small cell lung cancer by regulating KPNB1-mediated PD-L1 nuclear translocation
  29. Phytol-mixed micelles alleviate dexamethasone-induced osteoporosis in zebrafish: Activation of the MMP3–OPN–MAPK pathway-mediating bone remodeling
  30. Association between TGF-β1 and β-catenin expression in the vaginal wall of patients with pelvic organ prolapse
  31. Primary pleomorphic liposarcoma involving bilateral ovaries: Case report and literature review
  32. Effects of de novo donor-specific Class I and II antibodies on graft outcomes after liver transplantation: A pilot cohort study
  33. Sleep architecture in Alzheimer’s disease continuum: The deep sleep question
  34. Ephedra fragilis plant extract: A groundbreaking corrosion inhibitor for mild steel in acidic environments – electrochemical, EDX, DFT, and Monte Carlo studies
  35. Langerhans cell histiocytosis in an adult patient with upper jaw and pulmonary involvement: A case report
  36. Inhibition of mast cell activation by Jaranol-targeted Pirin ameliorates allergic responses in mouse allergic rhinitis
  37. Aeromonas veronii-induced septic arthritis of the hip in a child with acute lymphoblastic leukemia
  38. Clusterin activates the heat shock response via the PI3K/Akt pathway to protect cardiomyocytes from high-temperature-induced apoptosis
  39. Research progress on fecal microbiota transplantation in tumor prevention and treatment
  40. Low-pressure exposure influences the development of HAPE
  41. Stigmasterol alleviates endplate chondrocyte degeneration through inducing mitophagy by enhancing PINK1 mRNA acetylation via the ESR1/NAT10 axis
  42. AKAP12, mediated by transcription factor 21, inhibits cell proliferation, metastasis, and glycolysis in lung squamous cell carcinoma
  43. Association between PAX9 or MSX1 gene polymorphism and tooth agenesis risk: A meta-analysis
  44. A case of bloodstream infection caused by Neisseria gonorrhoeae
  45. Case of nasopharyngeal tuberculosis complicated with cervical lymph node and pulmonary tuberculosis
  46. p-Cymene inhibits pro-fibrotic and inflammatory mediators to prevent hepatic dysfunction
  47. GFPT2 promotes paclitaxel resistance in epithelial ovarian cancer cells via activating NF-κB signaling pathway
  48. Transfer RNA-derived fragment tRF-36 modulates varicose vein progression via human vascular smooth muscle cell Notch signaling
  49. RTA-408 attenuates the hepatic ischemia reperfusion injury in mice possibly by activating the Nrf2/HO-1 signaling pathway
  50. Decreased serum TIMP4 levels in patients with rheumatoid arthritis
  51. Sirt1 protects lupus nephritis by inhibiting the NLRP3 signaling pathway in human glomerular mesangial cells
  52. Sodium butyrate aids brain injury repair in neonatal rats
  53. Interaction of MTHFR polymorphism with PAX1 methylation in cervical cancer
  54. Convallatoxin inhibits proliferation and angiogenesis of glioma cells via regulating JAK/STAT3 pathway
  55. The effect of the PKR inhibitor, 2-aminopurine, on the replication of influenza A virus, and segment 8 mRNA splicing
  56. Effects of Ire1 gene on virulence and pathogenicity of Candida albicans
  57. Small cell lung cancer with small intestinal metastasis: Case report and literature review
  58. GRB14: A prognostic biomarker driving tumor progression in gastric cancer through the PI3K/AKT signaling pathway by interacting with COBLL1
  59. 15-Lipoxygenase-2 deficiency induces foam cell formation that can be restored by salidroside through the inhibition of arachidonic acid effects
  60. FTO alleviated the diabetic nephropathy progression by regulating the N6-methyladenosine levels of DACT1
  61. Clinical relevance of inflammatory markers in the evaluation of severity of ulcerative colitis: A retrospective study
  62. Zinc valproic acid complex promotes osteoblast differentiation and exhibits anti-osteoporotic potential
  63. Primary pulmonary synovial sarcoma in the bronchial cavity: A case report
  64. Metagenomic next-generation sequencing of alveolar lavage fluid improves the detection of pulmonary infection
  65. Uterine tumor resembling ovarian sex cord tumor with extensive rhabdoid differentiation: A case report
  66. Genomic analysis of a novel ST11(PR34365) Clostridioides difficile strain isolated from the human fecal of a CDI patient in Guizhou, China
  67. Effects of tiered cardiac rehabilitation on CRP, TNF-α, and physical endurance in older adults with coronary heart disease
  68. Changes in T-lymphocyte subpopulations in patients with colorectal cancer before and after acupoint catgut embedding acupuncture observation
  69. Modulating the tumor microenvironment: The role of traditional Chinese medicine in improving lung cancer treatment
  70. Alterations of metabolites related to microbiota–gut–brain axis in plasma of colon cancer, esophageal cancer, stomach cancer, and lung cancer patients
  71. Research on individualized drug sensitivity detection technology based on bio-3D printing technology for precision treatment of gastrointestinal stromal tumors
  72. CEBPB promotes ulcerative colitis-associated colorectal cancer by stimulating tumor growth and activating the NF-κB/STAT3 signaling pathway
  73. Oncolytic bacteria: A revolutionary approach to cancer therapy
  74. A de novo meningioma with rapid growth: A possible malignancy imposter?
  75. Diagnosis of secondary tuberculosis infection in an asymptomatic elderly with cancer using next-generation sequencing: Case report
  76. Hesperidin and its zinc(ii) complex enhance osteoblast differentiation and bone formation: In vitro and in vivo evaluations
  77. Research progress on the regulation of autophagy in cardiovascular diseases by chemokines
  78. Anti-arthritic, immunomodulatory, and inflammatory regulation by the benzimidazole derivative BMZ-AD: Insights from an FCA-induced rat model
  79. Immunoassay for pyruvate kinase M1/2 as an Alzheimer’s biomarker in CSF
  80. The role of HDAC11 in age-related hearing loss: Mechanisms and therapeutic implications
  81. Evaluation and application analysis of animal models of PIPNP based on data mining
  82. Therapeutic approaches for liver fibrosis/cirrhosis by targeting pyroptosis
  83. Fabrication of zinc oxide nanoparticles using Ruellia tuberosa leaf extract induces apoptosis through P53 and STAT3 signalling pathways in prostate cancer cells
  84. Haplo-hematopoietic stem cell transplantation and immunoradiotherapy for severe aplastic anemia complicated with nasopharyngeal carcinoma: A case report
  85. Modulation of the KEAP1-NRF2 pathway by Erianin: A novel approach to reduce psoriasiform inflammation and inflammatory signaling
  86. The expression of epidermal growth factor receptor 2 and its relationship with tumor-infiltrating lymphocytes and clinical pathological features in breast cancer patients
  87. Innovations in MALDI-TOF Mass Spectrometry: Bridging modern diagnostics and historical insights
  88. BAP1 complexes with YY1 and RBBP7 and its downstream targets in ccRCC cells
  89. Hypereosinophilic syndrome with elevated IgG4 and T-cell clonality: A report of two cases
  90. Electroacupuncture alleviates sciatic nerve injury in sciatica rats by regulating BDNF and NGF levels, myelin sheath degradation, and autophagy
  91. Polydatin prevents cholesterol gallstone formation by regulating cholesterol metabolism via PPAR-γ signaling
  92. RNF144A and RNF144B: Important molecules for health
  93. Analysis of the detection rate and related factors of thyroid nodules in the healthy population
  94. Artesunate inhibits hepatocellular carcinoma cell migration and invasion through OGA-mediated O-GlcNAcylation of ZEB1
  95. Endovascular management of post-pancreatectomy hemorrhage caused by a hepatic artery pseudoaneurysm: Case report and review of the literature
  96. Efficacy and safety of anti-PD-1/PD-L1 antibodies in patients with relapsed refractory diffuse large B-cell lymphoma: A meta-analysis
  97. SATB2 promotes humeral fracture healing in rats by activating the PI3K/AKT pathway
  98. Overexpression of the ferroptosis-related gene, NFS1, corresponds to gastric cancer growth and tumor immune infiltration
  99. Understanding risk factors and prognosis in diabetic foot ulcers
  100. Atractylenolide I alleviates the experimental allergic response in mice by suppressing TLR4/NF-kB/NLRP3 signalling
  101. FBXO31 inhibits the stemness characteristics of CD147 (+) melanoma stem cells
  102. Immune molecule diagnostics in colorectal cancer: CCL2 and CXCL11
  103. Inhibiting CXCR6 promotes senescence of activated hepatic stellate cells with limited proinflammatory SASP to attenuate hepatic fibrosis
  104. Cadmium toxicity, health risk and its remediation using low-cost biochar adsorbents
  105. Pulmonary cryptococcosis with headache as the first presentation: A case report
  106. Solitary pulmonary metastasis with cystic airspaces in colon cancer: A rare case report
  107. RUNX1 promotes denervation-induced muscle atrophy by activating the JUNB/NF-κB pathway and driving M1 macrophage polarization
  108. Morphometric analysis and immunobiological investigation of Indigofera oblongifolia on the infected lung with Plasmodium chabaudi
  109. The NuA4/TIP60 histone-modifying complex and Hr78 modulate the Lobe2 mutant eye phenotype
  110. Experimental study on salmon demineralized bone matrix loaded with recombinant human bone morphogenetic protein-2: In vitro and in vivo study
  111. A case of IgA nephropathy treated with a combination of telitacicept and half-dose glucocorticoids
  112. Analgesic and toxicological evaluation of cannabidiol-rich Moroccan Cannabis sativa L. (Khardala variety) extract: Evidence from an in vivo and in silico study
  113. Wound healing and signaling pathways
  114. Combination of immunotherapy and whole-brain radiotherapy on prognosis of patients with multiple brain metastases: A retrospective cohort study
  115. To explore the relationship between endometrial hyperemia and polycystic ovary syndrome
  116. Research progress on the impact of curcumin on immune responses in breast cancer
  117. Biogenic Cu/Ni nanotherapeutics from Descurainia sophia (L.) Webb ex Prantl seeds for the treatment of lung cancer
  118. Dapagliflozin attenuates atrial fibrosis via the HMGB1/RAGE pathway in atrial fibrillation rats
  119. Glycitein alleviates inflammation and apoptosis in keratinocytes via ROS-associated PI3K–Akt signalling pathway
  120. ADH5 inhibits proliferation but promotes EMT in non-small cell lung cancer cell through activating Smad2/Smad3
  121. Apoptotic efficacies of AgNPs formulated by Syzygium aromaticum leaf extract on 32D-FLT3-ITD human leukemia cell line with PI3K/AKT/mTOR signaling pathway
  122. Novel cuproptosis-related genes C1QBP and PFKP identified as prognostic and therapeutic targets in lung adenocarcinoma
  123. Bee venom promotes exosome secretion and alters miRNA cargo in T cells
  124. Treatment of pure red cell aplasia in a chronic kidney disease patient with roxadustat: A case report
  125. Comparative bioinformatics analysis of the Wnt pathway in breast cancer: Selection of novel biomarker panels associated with ER status
  126. Kynurenine facilitates renal cell carcinoma progression by suppressing M2 macrophage pyroptosis through inhibition of CASP1 cleavage
  127. RFX5 promotes the growth, motility, and inhibits apoptosis of gastric adenocarcinoma cells through the SIRT1/AMPK axis
  128. ALKBH5 exacerbates early cardiac damage after radiotherapy for breast cancer via m6A demethylation of TLR4
  129. Phytochemicals of Roman chamomile: Antioxidant, anti-aging, and whitening activities of distillation residues
  130. Circadian gene Cry1 inhibits the tumorigenicity of hepatocellular carcinoma by the BAX/BCL2-mediated apoptosis pathway
  131. The TNFR-RIPK1/RIPK3 signalling pathway mediates the effect of lanthanum on necroptosis of nerve cells
  132. Longitudinal monitoring of autoantibody dynamics in patients with early-stage non-small-cell lung cancer undergoing surgery
  133. The potential role of rutin, a flavonoid, in the management of cancer through modulation of cell signaling pathways
  134. Construction of pectinase gene engineering microbe and its application in tobacco sheets
  135. Construction of a microbial abundance prognostic scoring model based on intratumoral microbial data for predicting the prognosis of lung squamous cell carcinoma
  136. Sepsis complicated by haemophagocytic lymphohistiocytosis triggered by methicillin-resistant Staphylococcus aureus and human herpesvirus 8 in an immunocompromised elderly patient: A case report
  137. Sarcopenia in liver transplantation: A comprehensive bibliometric study of current research trends and future directions
  138. Advances in cancer immunotherapy and future directions in personalized medicine
  139. Can coronavirus disease 2019 affect male fertility or cause spontaneous abortion? A two-sample Mendelian randomization analysis
  140. Heat stroke associated with novel leukaemia inhibitory factor receptor gene variant in a Chinese infant
  141. PSME2 exacerbates ulcerative colitis by disrupting intestinal barrier function and promoting autophagy-dependent inflammation
  142. Hyperosmolar hyperglycemic state with severe hypernatremia coexisting with central diabetes insipidus: A case report and literature review
  143. Efficacy and mechanism of escin in improving the tissue microenvironment of blood vessel walls via anti-inflammatory and anticoagulant effects: Implications for clinical practice
  144. Merkel cell carcinoma: Clinicopathological analysis of three patients and literature review
  145. Genetic variants in VWF exon 26 and their implications for type 1 Von Willebrand disease in a Saudi Arabian population
  146. Lipoxin A4 improves myocardial ischemia/reperfusion injury through the Notch1-Nrf2 signaling pathway
  147. High levels of EPHB2 expression predict a poor prognosis and promote tumor progression in endometrial cancer
  148. Knockdown of SHP-2 delays renal tubular epithelial cell injury in diabetic nephropathy by inhibiting NLRP3 inflammasome-mediated pyroptosis
  149. Exploring the toxicity mechanisms and detoxification methods of Rhizoma Paridis
  150. Concomitant gastric carcinoma and primary hepatic angiosarcoma in a patient: A case report
  151. YAP1 inhibition protects retinal vascular endothelial cells under high glucose by inhibiting autophagy
  152. Identification of secretory protein related biomarkers for primary biliary cholangitis based on machine learning and experimental validation
  153. Integrated genomic and clinical modeling for prognostic assessment of radiotherapy response in rectal neoplasms
  154. Stem cell-based approaches for glaucoma treatment: a mini review
  155. Bacteriophage titering by optical density means: KOTE assays
  156. Neutrophil-related signature characterizes immune landscape and predicts prognosis of esophageal squamous cell carcinoma
  157. Integrated bioinformatic analysis and machine learning strategies to identify new potential immune biomarkers for Alzheimer’s disease and their targeting prediction with geniposide
  158. TRIM21 accelerates ferroptosis in intervertebral disc degeneration by promoting SLC7A11 ubiquitination and degradation
  159. TRIM21 accelerates ferroptosis in intervertebral disc degeneration by promoting SLC7A11 ubiquitination and degradation
  160. Histone modification and non-coding RNAs in skin aging: emerging therapeutic avenues
  161. A multiplicative behavioral model of DNA replication initiation in cells
  162. Biogenic gold nanoparticles synthesized from Pergularia daemia leaves: a novel approach for nasopharyngeal carcinoma therapy
  163. Creutzfeldt-Jakob disease mimicking Hashimoto’s encephalopathy: steroid response followed by decline
  164. Impact of semaphorin, Sema3F, on the gene transcription and protein expression of CREB and its binding protein CREBBP in primary hippocampal neurons of rats
  165. Iron overloaded M0 macrophages regulate hematopoietic stem cell proliferation and senescence via the Nrf2/Keap1/HO-1 pathway
  166. Revisiting the link between NADPH oxidase p22phox C242T polymorphism and ischemic stroke risk: an updated meta-analysis
  167. Exercise training preferentially modulates α1D-adrenergic receptor expression in peripheral arteries of hypertensive rats
  168. Overexpression of HE4/WFDC2 gene in mice leads to keratitis and corneal opacity
  169. Tumoral calcinosis complicating CKD-MBD in hemodialysis: a case report
  170. Mechanism of KLF4 Inhibition of epithelial-mesenchymal transition in gastric cancer cells
  171. Dissecting the molecular mechanisms of T cell infiltration in psoriatic lesions via cell-cell communication and regulatory network analysis
  172. Circadian rhythm-based prognostic features predict immune infiltration and tumor microenvironment in molecular subtypes of hepatocellular carcinoma
  173. Ecology and Environmental Science
  174. Optimization and comparative study of Bacillus consortia for cellulolytic potential and cellulase enzyme activity
  175. The complete mitochondrial genome analysis of Haemaphysalis hystricis Supino, 1897 (Ixodida: Ixodidae) and its phylogenetic implications
  176. Epidemiological characteristics and risk factors analysis of multidrug-resistant tuberculosis among tuberculosis population in Huzhou City, Eastern China
  177. Indices of human impacts on landscapes: How do they reflect the proportions of natural habitats?
  178. Genetic analysis of the Siberian flying squirrel population in the northern Changbai Mountains, Northeast China: Insights into population status and conservation
  179. Diversity and environmental drivers of Suillus communities in Pinus sylvestris var. mongolica forests of Inner Mongolia
  180. Global assessment of the fate of nitrogen deposition in forest ecosystems: Insights from 15N tracer studies
  181. Fungal and bacterial pathogenic co-infections mainly lead to the assembly of microbial community in tobacco stems
  182. Influencing of coal industry related airborne particulate matter on ocular surface tear film injury and inflammatory factor expression in Sprague-Dawley rats
  183. Temperature-dependent development, predation, and life table of Sphaerophoria macrogaster (Thomson) (Diptera: Syrphidae) feeding on Myzus persicae (Sulzer) (Homoptera: Aphididae)
  184. Eleonora’s falcon trophic interactions with insects within its breeding range: A systematic review
  185. Agriculture
  186. Integrated analysis of transcriptome, sRNAome, and degradome involved in the drought-response of maize Zhengdan958
  187. Variation in flower frost tolerance among seven apple cultivars and transcriptome response patterns in two contrastingly frost-tolerant selected cultivars
  188. Heritability of durable resistance to stripe rust in bread wheat (Triticum aestivum L.)
  189. Molecular mechanism of follicular development in laying hens based on the regulation of water metabolism
  190. Molecular identification and control studies on Coridius sp. (Hemiptera: Dinidoridae) in Al-Khamra, south of Jeddah, Saudi Arabia
  191. 10.1515/biol-2025-1218
  192. Animal Science
  193. Effect of sex ratio on the life history traits of an important invasive species, Spodoptera frugiperda
  194. Plant Sciences
  195. Hairpin in a haystack: In silico identification and characterization of plant-conserved microRNA in Rafflesiaceae
  196. Widely targeted metabolomics of different tissues in Rubus corchorifolius
  197. The complete chloroplast genome of Gerbera piloselloides (L.) Cass., 1820 (Carduoideae, Asteraceae) and its phylogenetic analysis
  198. Field trial to correlate mineral solubilization activity of Pseudomonas aeruginosa and biochemical content of groundnut plants
  199. Correlation analysis between semen routine parameters and sperm DNA fragmentation index in patients with semen non-liquefaction: A retrospective study
  200. Plasticity of the anatomical traits of Rhododendron L. (Ericaceae) leaves and its implications in adaptation to the plateau environment
  201. Effects of Piriformospora indica and arbuscular mycorrhizal fungus on growth and physiology of Moringa oleifera under low-temperature stress
  202. Effects of different sources of potassium fertiliser on yield, fruit quality and nutrient absorption in “Harward” kiwifruit (Actinidia deliciosa)
  203. Comparative efficiency and residue levels of spraying programs against powdery mildew in grape varieties
  204. The DREB7 transcription factor enhances salt tolerance in soybean plants under salt stress
  205. Using plant electrical signals of water hyacinth (Eichhornia crassipes) for water pollution monitoring
  206. Response of hybrid grapes (Vitis spp.) to two biotic stress factors and their seedlessness status
  207. Metabolomic profiling reveals systemic metabolic reprogramming in Alternaria alternata under salt stress
  208. Effects of mixed salinity and alkali stress on photosynthetic characteristics and PEPC gene expression of vegetable soybean seedlings
  209. Food Science
  210. Phytochemical analysis of Stachys iva: Discovering the optimal extract conditions and its bioactive compounds
  211. Review on role of honey in disease prevention and treatment through modulation of biological activities
  212. Computational analysis of polymorphic residues in maltose and maltotriose transporters of a wild Saccharomyces cerevisiae strain
  213. Optimization of phenolic compound extraction from Tunisian squash by-products: A sustainable approach for antioxidant and antibacterial applications
  214. Liupao tea aqueous extract alleviates dextran sulfate sodium-induced ulcerative colitis in rats by modulating the gut microbiota
  215. Toxicological qualities and detoxification trends of fruit by-products for valorization: A review
  216. Polyphenolic spectrum of cornelian cherry fruits and their health-promoting effect
  217. Optimizing the encapsulation of the refined extract of squash peels for functional food applications: A sustainable approach to reduce food waste
  218. Advancements in curcuminoid formulations: An update on bioavailability enhancement strategies curcuminoid bioavailability and formulations
  219. Impact of saline sprouting on antioxidant properties and bioactive compounds in chia seeds
  220. The dilemma of food genetics and improvement
  221. Causal effects of trace elements on congenital foot deformities and their subtypes: a Mendelian randomization study with gut microbiota mediation
  222. Honey meets acidity: a novel biopreservative approach against foodborne pathogens
  223. Bioengineering and Biotechnology
  224. Impact of hyaluronic acid-modified hafnium metalorganic frameworks containing rhynchophylline on Alzheimer’s disease
  225. Emerging patterns in nanoparticle-based therapeutic approaches for rheumatoid arthritis: A comprehensive bibliometric and visual analysis spanning two decades
  226. Application of CRISPR/Cas gene editing for infectious disease control in poultry
  227. Preparation of hafnium nitride-coated titanium implants by magnetron sputtering technology and evaluation of their antibacterial properties and biocompatibility
  228. Preparation and characterization of lemongrass oil nanoemulsion: Antimicrobial, antibiofilm, antioxidant, and anticancer activities
  229. Fluorescent detection of sialic acid–binding lectins using functionalized quantum dots in ELISA format
  230. Smart tectorigenin-loaded ZnO hydrogel nanocomposites for targeted wound healing: synthesis, characterization, and biological evaluation
  231. Corrigendum
  232. Corrigendum to “Utilization of convolutional neural networks to analyze microscopic images for high-throughput screening of mesenchymal stem cells”
  233. Corrigendum to “Effects of Ire1 gene on virulence and pathogenicity of Candida albicans”
  234. Retraction
  235. Retraction of “Down-regulation of miR-539 indicates poor prognosis in patients with pancreatic cancer”
https://doi.org/10.1515/biol-2025-1214
Keywords for this article
corneal transplantation; DZ2002; homocysteine; PI3K/AKT signaling pathway; rejection; TRAF1
Creative Commons
BY 4.0

Articles in the same Issue

  1. Safety assessment and modulation of hepatic CYP3A4 and UGT enzymes by Glycyrrhiza glabra aqueous extract in female Sprague–Dawley rats
  2. Adult-onset Still’s disease with hemophagocytic lymphohistiocytosis and minimal change disease
  3. Role of DZ2002 in reducing corneal graft rejection in rats by influencing Th17 activation via inhibition of the PI3K/AKT pathway and downregulation of TRAF1
  4. Biomedical Sciences
  5. Mechanism of triptolide regulating proliferation and apoptosis of hepatoma cells by inhibiting JAK/STAT pathway
  6. Maslinic acid improves mitochondrial function and inhibits oxidative stress and autophagy in human gastric smooth muscle cells
  7. Comparative analysis of inflammatory biomarkers for the diagnosis of neonatal sepsis: IL-6, IL-8, SAA, CRP, and PCT
  8. Post-pandemic insights on COVID-19 and premature ovarian insufficiency
  9. Proteome differences of dental stem cells between permanent and deciduous teeth by data-independent acquisition proteomics
  10. Optimizing a modified cetyltrimethylammonium bromide protocol for fungal DNA extraction: Insights from multilocus gene amplification
  11. Preliminary analysis of the role of small hepatitis B surface proteins mutations in the pathogenesis of occult hepatitis B infection via the endoplasmic reticulum stress-induced UPR-ERAD pathway
  12. Efficacy of alginate-coated gold nanoparticles against antibiotics-resistant Staphylococcus and Streptococcus pathogens of acne origins
  13. Battling COVID-19 leveraging nanobiotechnology: Gold and silver nanoparticle–B-escin conjugates as SARS-CoV-2 inhibitors
  14. Neurodegenerative diseases and neuroinflammation-induced apoptosis
  15. Impact of fracture fixation surgery on cognitive function and the gut microbiota in mice with a history of stroke
  16. COLEC10: A potential tumor suppressor and prognostic biomarker in hepatocellular carcinoma through modulation of EMT and PI3K-AKT pathways
  17. High-temperature requirement serine protease A2 inhibitor UCF-101 ameliorates damaged neurons in traumatic brain-injured rats by the AMPK/NF-κB pathway
  18. SIK1 inhibits IL-1β-stimulated cartilage apoptosis and inflammation in vitro through the CRTC2/CREB1 signaling
  19. Rutin–chitooligosaccharide complex: Comprehensive evaluation of its anti-inflammatory and analgesic properties in vitro and in vivo
  20. Knockdown of Aurora kinase B alleviates high glucose-triggered trophoblast cells damage and inflammation during gestational diabetes
  21. Calcium-sensing receptors promoted Homer1 expression and osteogenic differentiation in bone marrow mesenchymal stem cells
  22. ABI3BP can inhibit the proliferation, invasion, and epithelial–mesenchymal transition of non-small-cell lung cancer cells
  23. Changes in blood glucose and metabolism in hyperuricemia mice
  24. Rapid detection of the GJB2 c.235delC mutation based on CRISPR-Cas13a combined with lateral flow dipstick
  25. IL-11 promotes Ang II-induced autophagy inhibition and mitochondrial dysfunction in atrial fibroblasts
  26. Short-chain fatty acid attenuates intestinal inflammation by regulation of gut microbial composition in antibiotic-associated diarrhea
  27. Application of metagenomic next-generation sequencing in the diagnosis of pathogens in patients with diabetes complicated by community-acquired pneumonia
  28. NAT10 promotes radiotherapy resistance in non-small cell lung cancer by regulating KPNB1-mediated PD-L1 nuclear translocation
  29. Phytol-mixed micelles alleviate dexamethasone-induced osteoporosis in zebrafish: Activation of the MMP3–OPN–MAPK pathway-mediating bone remodeling
  30. Association between TGF-β1 and β-catenin expression in the vaginal wall of patients with pelvic organ prolapse
  31. Primary pleomorphic liposarcoma involving bilateral ovaries: Case report and literature review
  32. Effects of de novo donor-specific Class I and II antibodies on graft outcomes after liver transplantation: A pilot cohort study
  33. Sleep architecture in Alzheimer’s disease continuum: The deep sleep question
  34. Ephedra fragilis plant extract: A groundbreaking corrosion inhibitor for mild steel in acidic environments – electrochemical, EDX, DFT, and Monte Carlo studies
  35. Langerhans cell histiocytosis in an adult patient with upper jaw and pulmonary involvement: A case report
  36. Inhibition of mast cell activation by Jaranol-targeted Pirin ameliorates allergic responses in mouse allergic rhinitis
  37. Aeromonas veronii-induced septic arthritis of the hip in a child with acute lymphoblastic leukemia
  38. Clusterin activates the heat shock response via the PI3K/Akt pathway to protect cardiomyocytes from high-temperature-induced apoptosis
  39. Research progress on fecal microbiota transplantation in tumor prevention and treatment
  40. Low-pressure exposure influences the development of HAPE
  41. Stigmasterol alleviates endplate chondrocyte degeneration through inducing mitophagy by enhancing PINK1 mRNA acetylation via the ESR1/NAT10 axis
  42. AKAP12, mediated by transcription factor 21, inhibits cell proliferation, metastasis, and glycolysis in lung squamous cell carcinoma
  43. Association between PAX9 or MSX1 gene polymorphism and tooth agenesis risk: A meta-analysis
  44. A case of bloodstream infection caused by Neisseria gonorrhoeae
  45. Case of nasopharyngeal tuberculosis complicated with cervical lymph node and pulmonary tuberculosis
  46. p-Cymene inhibits pro-fibrotic and inflammatory mediators to prevent hepatic dysfunction
  47. GFPT2 promotes paclitaxel resistance in epithelial ovarian cancer cells via activating NF-κB signaling pathway
  48. Transfer RNA-derived fragment tRF-36 modulates varicose vein progression via human vascular smooth muscle cell Notch signaling
  49. RTA-408 attenuates the hepatic ischemia reperfusion injury in mice possibly by activating the Nrf2/HO-1 signaling pathway
  50. Decreased serum TIMP4 levels in patients with rheumatoid arthritis
  51. Sirt1 protects lupus nephritis by inhibiting the NLRP3 signaling pathway in human glomerular mesangial cells
  52. Sodium butyrate aids brain injury repair in neonatal rats
  53. Interaction of MTHFR polymorphism with PAX1 methylation in cervical cancer
  54. Convallatoxin inhibits proliferation and angiogenesis of glioma cells via regulating JAK/STAT3 pathway
  55. The effect of the PKR inhibitor, 2-aminopurine, on the replication of influenza A virus, and segment 8 mRNA splicing
  56. Effects of Ire1 gene on virulence and pathogenicity of Candida albicans
  57. Small cell lung cancer with small intestinal metastasis: Case report and literature review
  58. GRB14: A prognostic biomarker driving tumor progression in gastric cancer through the PI3K/AKT signaling pathway by interacting with COBLL1
  59. 15-Lipoxygenase-2 deficiency induces foam cell formation that can be restored by salidroside through the inhibition of arachidonic acid effects
  60. FTO alleviated the diabetic nephropathy progression by regulating the N6-methyladenosine levels of DACT1
  61. Clinical relevance of inflammatory markers in the evaluation of severity of ulcerative colitis: A retrospective study
  62. Zinc valproic acid complex promotes osteoblast differentiation and exhibits anti-osteoporotic potential
  63. Primary pulmonary synovial sarcoma in the bronchial cavity: A case report
  64. Metagenomic next-generation sequencing of alveolar lavage fluid improves the detection of pulmonary infection
  65. Uterine tumor resembling ovarian sex cord tumor with extensive rhabdoid differentiation: A case report
  66. Genomic analysis of a novel ST11(PR34365) Clostridioides difficile strain isolated from the human fecal of a CDI patient in Guizhou, China
  67. Effects of tiered cardiac rehabilitation on CRP, TNF-α, and physical endurance in older adults with coronary heart disease
  68. Changes in T-lymphocyte subpopulations in patients with colorectal cancer before and after acupoint catgut embedding acupuncture observation
  69. Modulating the tumor microenvironment: The role of traditional Chinese medicine in improving lung cancer treatment
  70. Alterations of metabolites related to microbiota–gut–brain axis in plasma of colon cancer, esophageal cancer, stomach cancer, and lung cancer patients
  71. Research on individualized drug sensitivity detection technology based on bio-3D printing technology for precision treatment of gastrointestinal stromal tumors
  72. CEBPB promotes ulcerative colitis-associated colorectal cancer by stimulating tumor growth and activating the NF-κB/STAT3 signaling pathway
  73. Oncolytic bacteria: A revolutionary approach to cancer therapy
  74. A de novo meningioma with rapid growth: A possible malignancy imposter?
  75. Diagnosis of secondary tuberculosis infection in an asymptomatic elderly with cancer using next-generation sequencing: Case report
  76. Hesperidin and its zinc(ii) complex enhance osteoblast differentiation and bone formation: In vitro and in vivo evaluations
  77. Research progress on the regulation of autophagy in cardiovascular diseases by chemokines
  78. Anti-arthritic, immunomodulatory, and inflammatory regulation by the benzimidazole derivative BMZ-AD: Insights from an FCA-induced rat model
  79. Immunoassay for pyruvate kinase M1/2 as an Alzheimer’s biomarker in CSF
  80. The role of HDAC11 in age-related hearing loss: Mechanisms and therapeutic implications
  81. Evaluation and application analysis of animal models of PIPNP based on data mining
  82. Therapeutic approaches for liver fibrosis/cirrhosis by targeting pyroptosis
  83. Fabrication of zinc oxide nanoparticles using Ruellia tuberosa leaf extract induces apoptosis through P53 and STAT3 signalling pathways in prostate cancer cells
  84. Haplo-hematopoietic stem cell transplantation and immunoradiotherapy for severe aplastic anemia complicated with nasopharyngeal carcinoma: A case report
  85. Modulation of the KEAP1-NRF2 pathway by Erianin: A novel approach to reduce psoriasiform inflammation and inflammatory signaling
  86. The expression of epidermal growth factor receptor 2 and its relationship with tumor-infiltrating lymphocytes and clinical pathological features in breast cancer patients
  87. Innovations in MALDI-TOF Mass Spectrometry: Bridging modern diagnostics and historical insights
  88. BAP1 complexes with YY1 and RBBP7 and its downstream targets in ccRCC cells
  89. Hypereosinophilic syndrome with elevated IgG4 and T-cell clonality: A report of two cases
  90. Electroacupuncture alleviates sciatic nerve injury in sciatica rats by regulating BDNF and NGF levels, myelin sheath degradation, and autophagy
  91. Polydatin prevents cholesterol gallstone formation by regulating cholesterol metabolism via PPAR-γ signaling
  92. RNF144A and RNF144B: Important molecules for health
  93. Analysis of the detection rate and related factors of thyroid nodules in the healthy population
  94. Artesunate inhibits hepatocellular carcinoma cell migration and invasion through OGA-mediated O-GlcNAcylation of ZEB1
  95. Endovascular management of post-pancreatectomy hemorrhage caused by a hepatic artery pseudoaneurysm: Case report and review of the literature
  96. Efficacy and safety of anti-PD-1/PD-L1 antibodies in patients with relapsed refractory diffuse large B-cell lymphoma: A meta-analysis
  97. SATB2 promotes humeral fracture healing in rats by activating the PI3K/AKT pathway
  98. Overexpression of the ferroptosis-related gene, NFS1, corresponds to gastric cancer growth and tumor immune infiltration
  99. Understanding risk factors and prognosis in diabetic foot ulcers
  100. Atractylenolide I alleviates the experimental allergic response in mice by suppressing TLR4/NF-kB/NLRP3 signalling
  101. FBXO31 inhibits the stemness characteristics of CD147 (+) melanoma stem cells
  102. Immune molecule diagnostics in colorectal cancer: CCL2 and CXCL11
  103. Inhibiting CXCR6 promotes senescence of activated hepatic stellate cells with limited proinflammatory SASP to attenuate hepatic fibrosis
  104. Cadmium toxicity, health risk and its remediation using low-cost biochar adsorbents
  105. Pulmonary cryptococcosis with headache as the first presentation: A case report
  106. Solitary pulmonary metastasis with cystic airspaces in colon cancer: A rare case report
  107. RUNX1 promotes denervation-induced muscle atrophy by activating the JUNB/NF-κB pathway and driving M1 macrophage polarization
  108. Morphometric analysis and immunobiological investigation of Indigofera oblongifolia on the infected lung with Plasmodium chabaudi
  109. The NuA4/TIP60 histone-modifying complex and Hr78 modulate the Lobe2 mutant eye phenotype
  110. Experimental study on salmon demineralized bone matrix loaded with recombinant human bone morphogenetic protein-2: In vitro and in vivo study
  111. A case of IgA nephropathy treated with a combination of telitacicept and half-dose glucocorticoids
  112. Analgesic and toxicological evaluation of cannabidiol-rich Moroccan Cannabis sativa L. (Khardala variety) extract: Evidence from an in vivo and in silico study
  113. Wound healing and signaling pathways
  114. Combination of immunotherapy and whole-brain radiotherapy on prognosis of patients with multiple brain metastases: A retrospective cohort study
  115. To explore the relationship between endometrial hyperemia and polycystic ovary syndrome
  116. Research progress on the impact of curcumin on immune responses in breast cancer
  117. Biogenic Cu/Ni nanotherapeutics from Descurainia sophia (L.) Webb ex Prantl seeds for the treatment of lung cancer
  118. Dapagliflozin attenuates atrial fibrosis via the HMGB1/RAGE pathway in atrial fibrillation rats
  119. Glycitein alleviates inflammation and apoptosis in keratinocytes via ROS-associated PI3K–Akt signalling pathway
  120. ADH5 inhibits proliferation but promotes EMT in non-small cell lung cancer cell through activating Smad2/Smad3
  121. Apoptotic efficacies of AgNPs formulated by Syzygium aromaticum leaf extract on 32D-FLT3-ITD human leukemia cell line with PI3K/AKT/mTOR signaling pathway
  122. Novel cuproptosis-related genes C1QBP and PFKP identified as prognostic and therapeutic targets in lung adenocarcinoma
  123. Bee venom promotes exosome secretion and alters miRNA cargo in T cells
  124. Treatment of pure red cell aplasia in a chronic kidney disease patient with roxadustat: A case report
  125. Comparative bioinformatics analysis of the Wnt pathway in breast cancer: Selection of novel biomarker panels associated with ER status
  126. Kynurenine facilitates renal cell carcinoma progression by suppressing M2 macrophage pyroptosis through inhibition of CASP1 cleavage
  127. RFX5 promotes the growth, motility, and inhibits apoptosis of gastric adenocarcinoma cells through the SIRT1/AMPK axis
  128. ALKBH5 exacerbates early cardiac damage after radiotherapy for breast cancer via m6A demethylation of TLR4
  129. Phytochemicals of Roman chamomile: Antioxidant, anti-aging, and whitening activities of distillation residues
  130. Circadian gene Cry1 inhibits the tumorigenicity of hepatocellular carcinoma by the BAX/BCL2-mediated apoptosis pathway
  131. The TNFR-RIPK1/RIPK3 signalling pathway mediates the effect of lanthanum on necroptosis of nerve cells
  132. Longitudinal monitoring of autoantibody dynamics in patients with early-stage non-small-cell lung cancer undergoing surgery
  133. The potential role of rutin, a flavonoid, in the management of cancer through modulation of cell signaling pathways
  134. Construction of pectinase gene engineering microbe and its application in tobacco sheets
  135. Construction of a microbial abundance prognostic scoring model based on intratumoral microbial data for predicting the prognosis of lung squamous cell carcinoma
  136. Sepsis complicated by haemophagocytic lymphohistiocytosis triggered by methicillin-resistant Staphylococcus aureus and human herpesvirus 8 in an immunocompromised elderly patient: A case report
  137. Sarcopenia in liver transplantation: A comprehensive bibliometric study of current research trends and future directions
  138. Advances in cancer immunotherapy and future directions in personalized medicine
  139. Can coronavirus disease 2019 affect male fertility or cause spontaneous abortion? A two-sample Mendelian randomization analysis
  140. Heat stroke associated with novel leukaemia inhibitory factor receptor gene variant in a Chinese infant
  141. PSME2 exacerbates ulcerative colitis by disrupting intestinal barrier function and promoting autophagy-dependent inflammation
  142. Hyperosmolar hyperglycemic state with severe hypernatremia coexisting with central diabetes insipidus: A case report and literature review
  143. Efficacy and mechanism of escin in improving the tissue microenvironment of blood vessel walls via anti-inflammatory and anticoagulant effects: Implications for clinical practice
  144. Merkel cell carcinoma: Clinicopathological analysis of three patients and literature review
  145. Genetic variants in VWF exon 26 and their implications for type 1 Von Willebrand disease in a Saudi Arabian population
  146. Lipoxin A4 improves myocardial ischemia/reperfusion injury through the Notch1-Nrf2 signaling pathway
  147. High levels of EPHB2 expression predict a poor prognosis and promote tumor progression in endometrial cancer
  148. Knockdown of SHP-2 delays renal tubular epithelial cell injury in diabetic nephropathy by inhibiting NLRP3 inflammasome-mediated pyroptosis
  149. Exploring the toxicity mechanisms and detoxification methods of Rhizoma Paridis
  150. Concomitant gastric carcinoma and primary hepatic angiosarcoma in a patient: A case report
  151. YAP1 inhibition protects retinal vascular endothelial cells under high glucose by inhibiting autophagy
  152. Identification of secretory protein related biomarkers for primary biliary cholangitis based on machine learning and experimental validation
  153. Integrated genomic and clinical modeling for prognostic assessment of radiotherapy response in rectal neoplasms
  154. Stem cell-based approaches for glaucoma treatment: a mini review
  155. Bacteriophage titering by optical density means: KOTE assays
  156. Neutrophil-related signature characterizes immune landscape and predicts prognosis of esophageal squamous cell carcinoma
  157. Integrated bioinformatic analysis and machine learning strategies to identify new potential immune biomarkers for Alzheimer’s disease and their targeting prediction with geniposide
  158. TRIM21 accelerates ferroptosis in intervertebral disc degeneration by promoting SLC7A11 ubiquitination and degradation
  159. TRIM21 accelerates ferroptosis in intervertebral disc degeneration by promoting SLC7A11 ubiquitination and degradation
  160. Histone modification and non-coding RNAs in skin aging: emerging therapeutic avenues
  161. A multiplicative behavioral model of DNA replication initiation in cells
  162. Biogenic gold nanoparticles synthesized from Pergularia daemia leaves: a novel approach for nasopharyngeal carcinoma therapy
  163. Creutzfeldt-Jakob disease mimicking Hashimoto’s encephalopathy: steroid response followed by decline
  164. Impact of semaphorin, Sema3F, on the gene transcription and protein expression of CREB and its binding protein CREBBP in primary hippocampal neurons of rats
  165. Iron overloaded M0 macrophages regulate hematopoietic stem cell proliferation and senescence via the Nrf2/Keap1/HO-1 pathway
  166. Revisiting the link between NADPH oxidase p22phox C242T polymorphism and ischemic stroke risk: an updated meta-analysis
  167. Exercise training preferentially modulates α1D-adrenergic receptor expression in peripheral arteries of hypertensive rats
  168. Overexpression of HE4/WFDC2 gene in mice leads to keratitis and corneal opacity
  169. Tumoral calcinosis complicating CKD-MBD in hemodialysis: a case report
  170. Mechanism of KLF4 Inhibition of epithelial-mesenchymal transition in gastric cancer cells
  171. Dissecting the molecular mechanisms of T cell infiltration in psoriatic lesions via cell-cell communication and regulatory network analysis
  172. Circadian rhythm-based prognostic features predict immune infiltration and tumor microenvironment in molecular subtypes of hepatocellular carcinoma
  173. Ecology and Environmental Science
  174. Optimization and comparative study of Bacillus consortia for cellulolytic potential and cellulase enzyme activity
  175. The complete mitochondrial genome analysis of Haemaphysalis hystricis Supino, 1897 (Ixodida: Ixodidae) and its phylogenetic implications
  176. Epidemiological characteristics and risk factors analysis of multidrug-resistant tuberculosis among tuberculosis population in Huzhou City, Eastern China
  177. Indices of human impacts on landscapes: How do they reflect the proportions of natural habitats?
  178. Genetic analysis of the Siberian flying squirrel population in the northern Changbai Mountains, Northeast China: Insights into population status and conservation
  179. Diversity and environmental drivers of Suillus communities in Pinus sylvestris var. mongolica forests of Inner Mongolia
  180. Global assessment of the fate of nitrogen deposition in forest ecosystems: Insights from 15N tracer studies
  181. Fungal and bacterial pathogenic co-infections mainly lead to the assembly of microbial community in tobacco stems
  182. Influencing of coal industry related airborne particulate matter on ocular surface tear film injury and inflammatory factor expression in Sprague-Dawley rats
  183. Temperature-dependent development, predation, and life table of Sphaerophoria macrogaster (Thomson) (Diptera: Syrphidae) feeding on Myzus persicae (Sulzer) (Homoptera: Aphididae)
  184. Eleonora’s falcon trophic interactions with insects within its breeding range: A systematic review
  185. Agriculture
  186. Integrated analysis of transcriptome, sRNAome, and degradome involved in the drought-response of maize Zhengdan958
  187. Variation in flower frost tolerance among seven apple cultivars and transcriptome response patterns in two contrastingly frost-tolerant selected cultivars
  188. Heritability of durable resistance to stripe rust in bread wheat (Triticum aestivum L.)
  189. Molecular mechanism of follicular development in laying hens based on the regulation of water metabolism
  190. Molecular identification and control studies on Coridius sp. (Hemiptera: Dinidoridae) in Al-Khamra, south of Jeddah, Saudi Arabia
  191. 10.1515/biol-2025-1218
  192. Animal Science
  193. Effect of sex ratio on the life history traits of an important invasive species, Spodoptera frugiperda
  194. Plant Sciences
  195. Hairpin in a haystack: In silico identification and characterization of plant-conserved microRNA in Rafflesiaceae
  196. Widely targeted metabolomics of different tissues in Rubus corchorifolius
  197. The complete chloroplast genome of Gerbera piloselloides (L.) Cass., 1820 (Carduoideae, Asteraceae) and its phylogenetic analysis
  198. Field trial to correlate mineral solubilization activity of Pseudomonas aeruginosa and biochemical content of groundnut plants
  199. Correlation analysis between semen routine parameters and sperm DNA fragmentation index in patients with semen non-liquefaction: A retrospective study
  200. Plasticity of the anatomical traits of Rhododendron L. (Ericaceae) leaves and its implications in adaptation to the plateau environment
  201. Effects of Piriformospora indica and arbuscular mycorrhizal fungus on growth and physiology of Moringa oleifera under low-temperature stress
  202. Effects of different sources of potassium fertiliser on yield, fruit quality and nutrient absorption in “Harward” kiwifruit (Actinidia deliciosa)
  203. Comparative efficiency and residue levels of spraying programs against powdery mildew in grape varieties
  204. The DREB7 transcription factor enhances salt tolerance in soybean plants under salt stress
  205. Using plant electrical signals of water hyacinth (Eichhornia crassipes) for water pollution monitoring
  206. Response of hybrid grapes (Vitis spp.) to two biotic stress factors and their seedlessness status
  207. Metabolomic profiling reveals systemic metabolic reprogramming in Alternaria alternata under salt stress
  208. Effects of mixed salinity and alkali stress on photosynthetic characteristics and PEPC gene expression of vegetable soybean seedlings
  209. Food Science
  210. Phytochemical analysis of Stachys iva: Discovering the optimal extract conditions and its bioactive compounds
  211. Review on role of honey in disease prevention and treatment through modulation of biological activities
  212. Computational analysis of polymorphic residues in maltose and maltotriose transporters of a wild Saccharomyces cerevisiae strain
  213. Optimization of phenolic compound extraction from Tunisian squash by-products: A sustainable approach for antioxidant and antibacterial applications
  214. Liupao tea aqueous extract alleviates dextran sulfate sodium-induced ulcerative colitis in rats by modulating the gut microbiota
  215. Toxicological qualities and detoxification trends of fruit by-products for valorization: A review
  216. Polyphenolic spectrum of cornelian cherry fruits and their health-promoting effect
  217. Optimizing the encapsulation of the refined extract of squash peels for functional food applications: A sustainable approach to reduce food waste
  218. Advancements in curcuminoid formulations: An update on bioavailability enhancement strategies curcuminoid bioavailability and formulations
  219. Impact of saline sprouting on antioxidant properties and bioactive compounds in chia seeds
  220. The dilemma of food genetics and improvement
  221. Causal effects of trace elements on congenital foot deformities and their subtypes: a Mendelian randomization study with gut microbiota mediation
  222. Honey meets acidity: a novel biopreservative approach against foodborne pathogens
  223. Bioengineering and Biotechnology
  224. Impact of hyaluronic acid-modified hafnium metalorganic frameworks containing rhynchophylline on Alzheimer’s disease
  225. Emerging patterns in nanoparticle-based therapeutic approaches for rheumatoid arthritis: A comprehensive bibliometric and visual analysis spanning two decades
  226. Application of CRISPR/Cas gene editing for infectious disease control in poultry
  227. Preparation of hafnium nitride-coated titanium implants by magnetron sputtering technology and evaluation of their antibacterial properties and biocompatibility
  228. Preparation and characterization of lemongrass oil nanoemulsion: Antimicrobial, antibiofilm, antioxidant, and anticancer activities
  229. Fluorescent detection of sialic acid–binding lectins using functionalized quantum dots in ELISA format
  230. Smart tectorigenin-loaded ZnO hydrogel nanocomposites for targeted wound healing: synthesis, characterization, and biological evaluation
  231. Corrigendum
  232. Corrigendum to “Utilization of convolutional neural networks to analyze microscopic images for high-throughput screening of mesenchymal stem cells”
  233. Corrigendum to “Effects of Ire1 gene on virulence and pathogenicity of Candida albicans”
  234. Retraction
  235. Retraction of “Down-regulation of miR-539 indicates poor prognosis in patients with pancreatic cancer”
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Winner of the OpenAthens UX Award 2026
Winner of the OpenAthens UX Award 2026
Have an idea on how to improve our website?
Please write us.
© 2026 De Gruyter Brill
Downloaded on 23.3.2026 from https://www.degruyterbrill.com/document/doi/10.1515/biol-2025-1214/html
Scroll to top button