Abstract:
Objective To explore whether baseline plasma fibroblast growth factor 21 (FGF21) levels are associated with treatment outcomes in patients with dilated cardiomyopathy (DCM) who are overweight/obese. Methods A prospective study included 217 patients with DCM and overweight/obese (24 kg/m2≤ BMI < 40 kg/m2). All patients received standardized heart failure medication. Baseline FGF21 levels were measured at enrollment. Left ventricular ejection fraction (LVEF), NYHA functional class, and B-type natriuretic peptide changes were assessed during a 12- month follow-up. Patients were divided into an effective group (n=155) and an ineffective group (n=62) based on the criteria for treatment effectiveness (meeting≥2 criteria: LVEF improvement≥10%, NYHA functional class improvement≥1, and BNP decrease≥50%). Spearman correlation analysis, Logistic regression, and ROC curve analysis were employed to evaluate the association of FGF21 with therapeutic efficacy and major adverse cardiovascular events. Results Baseline FGF21 levels in the invalid group were higher than those in the valid group (median: 244.65 pg/mL vs 198.40 pg/mL, P<0.001). FGF21 was positively correlated with follow-up BNP (r=0.184, P<0.05) and negatively correlated with follow-up LVEF (r=-0.140, P<0.05). Multivariate Logistic regression analysis showed that FGF21 was an independent risk factor for poor treatment response (P<0.001). The area under the receiver operating characteristic curve (ROC) was 0.90, with the optimal cutoff value being 205 pg/mL. The high FGF21 group (≥205 pg/mL) had a higher risk of major adverse cardiovascular events (P<0.001). Conclusion Baseline FGF21 elevation is independently associated with reduced treatment response and increased risk of MACE in DCM patients with overweight/obesity, serving as a prognostic biomarker for predicting treatment efficacy, which facilitates risk stratification and personalized therapy.

Abstract:
Objective To investigate the effects ofa dual-functional cationic nucleic acid nanodelivery system (DCNA) loaded with small interfering RNA targeting Bruton’s tyrosine kinase (siBTK) on the immune-inflammatory microenvironment of periodontitis by regulating BTK expression. Methods DCNAsiBTK was prepared using a double-emulsion solvent evaporation method, and its physicochemical properties and sustained-release performance were characterized. Cytotoxicity was evaluated using the Cell Counting Kit-8 (CCK-8) assay. Antibacterial activity against periodontal pathogens was assessed by colony-forming assays. BTK mRNA and protein expression levels were determined by quantitative real-time PCR (qRT-PCR) and Western blotting, respectively. Enzyme-linked immunosorbent assay (ELISA) was performed to quantify the levels of inflammatory cytokines, including IL-1β, IL-6, and TNF-α . A murine periodontitis model was established and treated with DCNAsiBTK. Therapeutic efficacy and biosafety were evaluated using qPCR, micro-computed tomography (micro-CT), hematoxylin-eosin (HE) staining, and immunohistochemistry (IHC). Results Stable DCNAsiBTK nanoparticles with an average particle size of (168.8 ± 2.2) nm and a zeta potential of (17.9 ± 0.8) mV were successfully prepared. Transmission electron microscopy (TEM) revealed uniformly spherical morphology with favorable sustained-release properties. CCK-8 assays demonstrated no significant cytotoxicity of DCNAsiBTK within the concentration range of 0-100 nmol/L. In vitro studies showed that DCNAsiBTK effectively inhibited the growth of Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), and Fusobacterium nucleatum (Fn), indicating broad-spectrum antibacterial activity. qRT- PCR and Western blotting analyses demonstrated that DCNAsiBTK downregulated BTK mRNA and protein expression in a dose-dependent manner. ELISA results showed that DCNAsiBTK significantly reduced the secretion of IL-1β, IL-6, and TNF-α, thereby attenuating inflammatory responses. In vivo experiments further demonstrated that DCNAsiBTK suppressed anaerobic bacterial proliferation within periodontal pockets, alleviated alveolar bone resorption and periodontal inflammation, and caused no obvious pathological damage in major organs, including the heart, liver, spleen, lungs, and kidneys. Conclusion DCNAsiBTK efficiently encapsulated and released siBTK, and exhibited dual antibacterial and anti-inflammatory activities. By suppressing BTK expression, DCNAsiBTK effectively attenuated the progression of periodontitis, providing a promising novel strategy for adjunctive periodontal therapy.

Abstract:
Objective To investigate DERL1 protein expression in esophageal squamous cell carcinoma (ESCC) and its impact on prognosis, malignant behavior, and the immune microenvironment. Methods DERL1 expression in ESCC and adjacent tissues was detected by immunohistochemistry to analyze its clinicopathological and prognostic significance. After DERL1 knockdown in human esophageal cancer cell lines TE-1 and KYSE-450 cells, cell proliferation, colony formation, and migration were assessed via CCK-8, plate colony, and Transwell assays. The correlation between DERL1 and immune infiltration was analyzed using TIMER2.0 and validated by multiplex immunofluorescence. Results DERL1 protein was significantly upregulated in ESCC tissues. High DERL1 expression predicted shorter overall survival (Log-rank P=0.011) and served as an independent prognostic risk factor (HR=8.717, P=0.035). Functional cellular assays showed that DERL1 knockdown significantly inhibited proliferation, colony formation, and migration/invasion in both TE-1 and KYSE-450 cells (P<0.05). Bioinformatics analysis revealed that DERL1 expression was positively correlated with M0 macrophages and resting CD4+ memory T cells, while negatively correlated with CD8+T cells. Multiplex immunofluorescence indicated that regions with high DERL1 expression exhibited increased CD4+ but decreased CD8+ T cell infiltration. Conclusion DERL1 protein is highly expressed in ESCC and serves as an independent biomarker of poor prognosis. DERL1 promotes tumor cell proliferation and migration and is associated with an immunosuppressive tumor microenvironment. These findings suggest that DERL1 may serve as a potential therapeutic target for esophageal squamous cell carcinoma.

Abstract:
Objective To evaluate the surface properties, mechanical properties, biocompatibility, and ablity to inhibit protein and bacterial of the biomimetic superhydrophobic dental restorative resin material (BSDRM).Methods Nano ZrO2 was modified with fluorosilane (FAS) to obtain FAS-ZrO2, which, together with nano SiO2, was incorporated into a resin matrix composed of bisphenol A-glycerolate dimethacrylate (Bis-GMA), triethylene glycol dimethacrylate (TEGDMA), perfluoroalkyl acrylate (FMA), and a photoinitiator system to fabricate BSDRM. The chemical structure and surface morphology of the material were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), field-emission scanning electron microscopy (FE-SEM) and atomic force microscopy (AFM). Wettability parameters, including water contact angle, sliding angle and surface energy, were measured. Mechanical performance was assessed by Vickers hardness, diametral tensile strength and compressive strength tests. Biocompatibility was evaluated using CCK-8 assays and cell staining. The ability to inhibit protein adsorption and bacterial adhesion were examined by salivary mucin adsorption and Streptococcus mutans adhesion assays, respectively. Results Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) confirmed the successful grafting of FAS onto the ZrO2 surface. BSDRM exhibited a micro/nanoscale hierarchical roughness, with a water contact angle of 152.97°± 1.64°, a sliding angle of 7.37°± 1.91°, a surface energy of 1.20 ± 0.26 mN/m. Mechanical testing showed that BSDRM had significantly higher hardness than the hydrophilic resin and the commercial pit-and-fissure sealant, whereas its compressive strength was not significantly different from that of the sealant (P > 0.05). CCK-8 and cell staining results demonstrated that BSDRM was non-cytotoxic to L929 cells and preserved normal cellular morphology. In the protein adsorption assay, BSDRM showed the lowest mucin adsorption among all groups (P < 0.001). In the bacterial adhesion assay, the amount of S. mutans adhering to the BSDRM surface was significantly lower than that in the control groups (P < 0.01). Conclusions BSDRM was successfully developed in this study. The material combines excellent superhydrophobicity, favourable mechanical properties, good biocompatibility, and marked ability to inhibit protein adsorption and bacterial adhesion. By suppressing plaque formation, BSDRM may offer a promising new materials-based strategy for caries prevention.

Abstract:
Objective To investigate the regulatory mechanism of peptidylarginine deiminase 4 (PADI4) on matrix metalloproteinase (MMPs) expression and its potential role in the development of systemic lupus erythematosus (SLE). Methods ThemRNA expression levels of MMP-14/17/25 in SLE patients and healthy controls (HC) were measured by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR), and their serum protein levels were measured by enzyme-linked immunosorbent assay (ELISA). Spearman's rank correlation analysis was used to evaluate the correlations between MMPs levels and clinical parameters in SLE patients. Primary neutrophils were treated with the PADI4 inhibitor GSK484, or MMPs expression levels were examined in PADI4-knockdown neutrophil-like HL-60 (dHL-60) cells, and cell migration ability was assessed by wound healing assay. Results The mRNA expression levels of MMP- 14/17/25 in neutrophils from SLE patients were lower than those in the HC (all P<0.01). Serum levels of these three MMPs were also lower than those in the HC (all P<0.01). MMP-14 levels were negatively correlated with immunoglobulin G (IgG) (r=-0.266), immunoglobulin A (IgA) (r=- 0.286), and globulin (GLB)(r=-0.269) (all P<0.05), while both MMP-17 (r =-0.269) and MMP- 25(r =-0.244) levels were negatively correlated with urinary protein (UTP) (all P<0.05). After treatment with GSK484 (10 μmol/L) to inhibit PADI4 activity, the protein expression levels of MMP-14/17/25 in neutrophils were higher than those in the Vehicle group (all P<0.05), and the cell migration rate in wound healing assay was also greater than that in the Vehicle group (P<0.01). Furthermore, after knockdown of PADI4 expression, the mRNA expression levels of MMP- 14/17/25 were higher than those in the Vehicle group (all P<0.05). Conclusion MMP-14/17/25 are downregulated in SLE and negatively regulated by PADI4. PADI4 may contribute to SLE progression by suppressing MMPs expression and neutrophil migration. These findings suggest that MMPs may serve as novel biomarkers and therapeutic targets for SLE.

Abstract:
Pulmonary hypertension (PH) is a life-threatening condition characterized by a progressive elevation in pulmonary vascular pressure, ultimately leading to right ventricular dysfunction and failure. The core pathological features of PH include abnormal contraction of small pulmonary arteries and vascular remodeling. In recent years, endoplasmic reticulum stress (ERS), a critical adaptive response of cells to internal and external stressors, has been shown to play a key role in the development and progression of PH. Through the activation of major signaling pathways such as protein kinase R-like endoplasmic reticulum kinase (PERK), inositol-requiring enzyme 1α (IRE1α), and activating transcription factor 6 (ATF6), ERS is directly involved in regulating aberrant cellular behaviors and vascular remodeling in PH. This review systematically discusses the specific role of ERS in the pathogenesis of PH and evaluates its potential as a therapeutic target, aiming to provide a theoretical foundation for novel clinical interventions in PH.

Abstract:
Objective To investigate the expression of synaptotagmin 1 (SYT1) in hepatocellular carcinoma (HCC) and its clinical value. Methods Bioinformatics analysis was performed to predict the subcellular localization and expression of SYT1 in the tumor microenvironment (TME),as well as its biological functions in the progression of HCC. The expression of SYT1 in HCC tissues was verified by UALCAN, TIMER3.0 and HPA databases combined with IHC, and its correlation with MKI67 was analyzed. Combined with GEPIA2, TIMER3.0 databases and Rlanguage, the association of SYT1 expression level with the prognosis of HCC patients was explored, and survival analysis was carried out. Lentivirus-mediated SYT1 gene silencing was conducted, and the effects of SYT1 on the biological behaviors of HCC cells were explored by colony formation assay, EdU staining assay, cell scratch assay, and Transwell assay. The tumor-promoting effect of SYT1 was further validated in a nude mouse xenograft model. Results Bioinformatics prediction indicated that SYT1 is localized to the cell membrane and cytoplasm,and the expression of its family genes is mainly positively correlated. In HCC tissues, SYT1 expression was significantly upregulated (P=2.93×10-10) and positively correlated with MKI67 (ρs=0.419, P=3.3×10-17). Survival analysis showed that high SYT1 expression predicted poorer overall survival (P=0.015) and disease-specific survival (P=0.046 2) in HCC. Immune profiling analysis demonstrated that SYT1 was highly expressed in monocytes/macrophages, B cells and malignant cells in the TME. Enrichment analysis indicated that SYT1 may participate in multiple signaling pathways associated with HCC progression (P<0.05). IHC confirmed that the staining intensity of SYT1 was elevated in HCC tissues (P<0.000 1). Cellular assays showed that silencing SYT1 suppressed the proliferation, migration, and invasion of HCC cells (P<0.01). Animal experiments demonstrated that knockdown of SYT1 inhibited tumor growth in vivo (P<0.001).Conclusion SYT1 is upregulated in HCC and closely correlated with poor prognosis. It can promote the proliferation, migration and invasion of HCC cells as well as in vivo tumor growth, and is expected to become a potential novel target for the clinical treatment of HCC.

Abstract:
Objective To investigate the correlation between different female body mass index (BMI) values and key indicators of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment cycles, as well as their impact on obstetric outcomes. Methods A retrospective analysis was conducted on data from patients who underwent IVF/ICSI treatment, with 19 834 cycles included after propensity score matching to balance group differences. Based on the Chinese BMI classification standards with extended categories, patients were stratified into five groups: severely underweight (BMI<16), underweight (16.0 ≤BMI<18.5), normal weight (18.5≤BMI<24.0), overweight (24.0 ≤ BMI<28.0), and obese (BMI ≥ 28.0). Baseline characteristics, ovarian stimulation protocols, treatment processes, and assisted reproductive outcomes were compared among the groups. Additionally, 75 delivered patients with complications were followed up to analyze the association between BMI and pregnancy-related complications. Results No statistically significant differences were observed among the five groups in baseline parameters such as age, duration of infertility, or antral follicle count. Regarding ovarian stimulation protocols and processes, a statistically significant difference was found in the number of IVF oocytes retrieved (P<0.001), with the normal-weight group showing a significantly higher number than both the overweight and obese groups. In terms of assisted reproductive outcomes, no significant differences were observed in biochemical pregnancy rates or birth defect rates. However, cumulative live birth rates differed significantly among the groups (P<0.05), which was significantly higher in the overweight group than in the normal-weight group. Among the 75 followed-up patients, 25 were diagnosed with gestational hypertension, and 22 with gestational diabetes mellitus, of whom overweight and obese individuals accounted for 60.00% and 45.40% of the respective complications. Conclusion Female BMI is significantly associated with IVF/ICSI outcomes. Pre-pregnancy body mass evaluation should be emphasized in patients with extremely low body weight. Although the live birth rate of overweight and obese patients was comparable to that of the normal-weight group, weight reduction prior to treatment is strongly recommended to minimize risks of miscarriage, preterm birth, and pregnancy complications.

Abstract:
Objective To evaluate the impact of hysteroscopic examination and subsequent management on reproductive outcomes in embryo transfer cycles among patients undergoing preimplantation genetic testing (PGT). Methods This retrospective analysis included 261 transfer cycles from in vitro fertilization (IVF) and 267 transfer cycles from PGT. Among IVF cycles, 134 underwent hysteroscopy prior to embryo transfer (IVF-H group), while 127 did not (IVF-N group). Among PGT cycles, 140 received hysteroscopy before transfer (PGT-H group), and 127 did not (PGT-N group). We compared general characteristics and pregnancy outcomes between the IVF-H and IVF-N groups, as well as between the PGT-H and PGT-N groups. Results No significant differences were observed in maternal age, body mass index, or baseline hormone profiles between the IVF-H and IVF-N groups, nor between the PGT-H and PGT-N groups. The IVF-H group showed higher rates of biochemical pregnancy, clinical pregnancy, and live birth, along with a lower early miscarriage rate, compared with the IVF-N group. However, these differences did not reach statistical significance (all P>0.05). In the PGT cohort, the PGT-H group had significantly higher biochemical and clinical pregnancy rates than the PGT-N group (P<0.05). Early miscarriage rates were comparable between the PGT-H and PGT-N groups (P>0.05). Although the live birth rate was higher in the PGT-H group than in the PGT-N group, the difference was not statistically significant (P>0.05). Conclusion Hysteroscopic evaluation before embryo transfer may be associated with improved clinical pregnancy and live birth rates. This benefit appears more pronounced in patients undergoing PGT compared with those receiving conventional IVF treatment.

Abstract:
Objective To explore the effect of semaglutide (SEM) on myocardial injury induced by coronary microembolization (CME) in rats via the adenosine monophosphate-activated protein kinase (AMPK)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) signaling pathway. Methods Thirty-two rats were randomly assigned to four groups (n=8): the Sham group, the CME group, the CME + SEM group, and the CME + SEM + Compound C (CC, a specific inhibitor of AMPK) group. The CME model was established by injecting microembolic spheres into the left ventricle, while the Sham group received an equal volume of normal saline via left ventricular injection. The CME + SEM group received subcutaneous injections of SEM (1 mg/kg, once weekly) for 4 weeks before CME induction. Forthe CME + SEM + CC group, rats were treated with SEM for 4 weeks, followed by an intravenous injection of CC (0.25 mg/kg dissolved in sterile normal saline) 30 minutes before CME modeling. 24 hours post-modeling, all groups underwent the following assessments: echocardiography to evaluate cardiac function, HE staining for myocardial histopathological analysis, Heidenhain staining to quantify the microinfarct area, ELISA to determine serum cardiac troponin T (cTnT) levels, and Western blot analysis to detect the expression of proteins related to the AMPK/NLRP3 pathway in cardiac tissues. Results Compared with the Sham group, the CME group showed significantly deteriorated cardiac function, elevated serum cTnT levels, and a marked increase in myocardial microinfarct area (P<0.05). Compared with the CME group, the CME + SEM group exhibited significant improvements in cardiac function, decreased cTnT levels, and a remarkable reduction in myocardial microinfarct area (P<0.05). Histopathological analysis revealed a normal myocardial structure in the Sham group without infarction. In contrast, the CME group showed disorganized myocardial arrangement, accompanied by nuclear fragmentation and karyolysis. In the CME + SEM group, myocardial degeneration, edema and inflammatory cell infiltration were obviously alleviated. Compared with Sham group, the protein ratio-AMPK/AMPK slightly increased in the CME group, while the protein levels of NLRP3, gasdermin D-N domain (GSDMD-N), apoptosis-associated speck-like protein containing a CARD (ASC), caspase-1 p20, interleukin(IL)-1β , and IL-18 were markedly upregulated (P<0.05). Notably, compared to the CME group, the CME + SEM group had a further elevated p-AMPK/AMPK ratio, while the protein levels of NLRP3, GSDMD-N, ASC, caspase-1 p20, IL-1β and IL-18 were markedly decreased (P<0.05). Conclusion SEM significantly ameliorates CME- induced myocardial injury, and its mechanism of action may be closely associated with promoting AMPK phosphorylation, inhibiting NLRP3 inflammasome-mediated inflammatory responses, and suppressing pyroptosis.

Abstract:
Objective To investigate the effect of growth hormone (GH) pretreatment on the clinical outcomes of in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) in patients with diminished ovarian reserve (DOR). Methods A retrospective analysis was conducted on the clinical data of 359 DOR patients undergoing IVF/ICSI-ET. Based on the use of GH pretreatment before ovarian stimulation and/or its addition during the stimulation cycle, patients were divided into three groups: the Control group (n=119), receiving no GH pretreatment or addition; the Non-GH pretreatment group (n=115), receiving GH (4 IU/d) only during ovarian stimulation without pretreatment; and the GH pretreatment group (n=125), pretreated with GH (2 IU/d) for 4-6 weeks before stimulation and also receiving GH (4 IU/d) during stimulation. The basic characteristics, ovarian stimulation parameters, embryological data, and cumulative live birth rate (CLBR) were compared among the groups. Results ① No significant differences were found among the three groups regarding female age, male age, duration of infertility, body mass index (BMI), basal hormone levels, distribution of ovarian stimulation protocols, endometrial thickness, serum estradiol (E2) and luteinizing hormone (LH) levels on the day of HCG trigger, or the number of blastocysts transferred per cycle. ② The GH pretreatment group required significantly lower total gonadotropin (Gn) dosage and fewer days of stimulation than the Control and Non-GH pretreatment group(P<0.05). Compared to the Control group, the GH pretreatment group showed significantly higher numbers of retrieved oocytes, the number of mature oocytes, 2-pronuclei (2PN) zygotes, 2PN cleaved embryos, transferable embryos, high-quality embryos, as well as higher cumulative clinical pregnancy rates and CLBR (P<0.05). Compared to the Non-GH pretreatment group, the GH pretreatment group had significantly more transferable embryos, high-quality embryos, and a higher CLBR (P<0.05). ③ Compared to the Control group, the Non-GH pretreatment group only had a significantly higher number of retrieved oocytes (P<0.05), with no significant differences in the number of mature oocytes, 2PN fertilized oocytes, 2PN cleaved embryos, transferable blastocysts, high-quality blastocysts, cumulative clinical pregnancy rate, and CLBR. Conclusion For DOR patients, GH pretreatment for 4-6 weeks prior to ovarian stimulation can improve oocyte yield, embryo quality, cumulative clinical pregnancy rate, and CLBR, thereby enhancing the clinical outcomes of IVF/ICSI-ET.

Abstract:
Objective To investigate the effect of kynurenine-3-monooxygenase (KMO) on the function of fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA). Methods KMO expression in synovial tissues from RA patients and healthy controls (HC) was analyzed using bioinformatics. Synovial tissues from RA patients and HC were collected, and KMO expression in FLS within synovial tissues was detected by immunofluorescence (IF). In vitro experiments were performed using human rheumatoid arthritis fibroblast-like synoviocytes (MH7A). Following KMO overexpression, KMO knockdown, and KMO inhibition with the specific inhibitor Ro61-8048 in MH7A cells, Western blot, CCK-8, EdU, Transwell and ELISA assays were used to evaluate alterations in cell viability, proliferation, migration and secretory function. Results Bioinformatic analysis revealed significantly higher KMO gene expression in RA synovial tissue compared to the HC group (P<0.001). IF results showed increased KMO expression in FLS from RA synovial tissues relative to the HC group (P<0.01). Overexpression of KMO led to enhanced viability, proliferation, migration, and secretory capabilities of MH7A cells (all P<0.05), whereas knockdown or inhibition of KMO with Ro61-8048 markedly suppressed these functions (all P<0.05). Conclusion The expression ofKMO is upregulated in the synovial tissues of RA patients and promotes the activation of RA-FLS, suggesting that KMO may serve as a potential therapeutic target for RA treatment.

Abstract:
Objective To investigate molecular alterations in tumor cells of multiple myeloma (MM) before and after B-cell maturation antigen (BCMA)-directed chimeric antigen receptor T-cell (CAR- T) therapy, with a focus on changes in invariant chain (CD74) expression and its potential association with macrophage migration inhibitory factor (MIF). Methods Paired bone marrow samples collected before BCMA CAR-T infusion and at ≥3 months after infusion were analyzed. Differential gene expression, functional enrichment, and cell-cell communication analyses were performed in malignant plasma cell-related populations. In vitro coculture experiments and MIF stimulation assays were conducted using the U266 cell line, and cell proliferation was assessed by CCK-8 and EdU incorporation assays. Results CD74 expression was upregulated in MM cells at ≥3 months after BCMA CAR-T therapy, and CD74 expression was consistently higher in post- treatment samples than in paired pre-treatment samples across all four cases. Genes upregulated after treatment were mainly enriched in antigen processing and presentation, cell adhesion, and immune-related processes. Cell-cell communication analysis revealed prominent interaction signals involving MIF-related pathways. In vitro experiments showed that coculture conditions induced changes in membrane CD74 expression in tumor cells, and MIF stimulation promoted proliferation of U266 cells. Conclusion CD74 expression is upregulated in tumor-associated MM cells following BCMA CAR-T therapy and is accompanied by the activation of the MIF-CD74 axis, suggesting a potential role in post-treatment adaptive changes and warranting further investigation into its therapeutic relevance.

Abstract:
Objective To obtain stably transfected chimeric antigen receptor macrophages (CAR-M) targeting disialosylganglioside (GD2) based on the constitutive expression of myeloid differentiation factor 88 (MYD88). MethodsAn Anti-GD2 CAR adenoviral vector incorporating the CD3ζ (cluster of differentiation 3 zeta chain)-MYD88 fusion construct was designed and constructed. THP-1-derived macrophages (THP-1M), induced by phorbol 12-myristate 13-acetate (PMA) differentiation, were infected with this adenovirus to generate Anti-GD2 CAR THP-1 cells bearing a green fluorescent protein (GFP) tag. CAR expression was monitored using a live-cell imaging system. The phagocytic function of Empty CAR-M, Anti-GD2 CAR-M(CD3ζ), and Anti-GD2 CAR-M(CD3ζ-MYD88) groups against glioma cells was evaluated in vitro through co-culture experiments, and their polarization status was also analyzed. Results CAR adenovirus successfully transfected THP-1 to obtain Anti-GD2 CAR-M. After co-culture with glioma cells, flow cytometry results showed that compared with the Empty CAR-M group and the Anti-GD2 CAR-M(CD3ζ) group, the Anti-GD2 CAR-M(CD3ζ-MYD88) group exhibited enhanced phagocytosis against T98G cells with high GD2 expression (P<0.05) and also showed upregulation of CD86 (P<0.05). Compared with the Empty CAR-M group and the Anti-GD2 CAR-M(CD3ζ) group, the Anti-GD2 CAR-M(CD3ζ-MYD88)CAR-M group exhibited enhanced phagocytosis against T98G cells with high GD2 expression (P<0.05) and also showed upregulation of CD86 (P<0.05), and the expression of inflammatory cytokines interleukin (IL)-6 and IL-12 was increased in this group (P<0.05). Conclusion This study successfully constructed Anti-GD2 CAR-M cells and verified their phagocytic activity, demonstrating that constitutive expression of MYD88 can promote inflammatory polarization of CAR-M. This provides a potential structural modification approach for the development of pro-inflammatory CAR-M.

Abstract:
Retinal degeneration (RD) encompasses a group of disorders marked by progressive apoptosis of retinal cells and loss of retinal structural integrity, ultimately resulting in blindness. In recent years, the number of individuals affected by RD-related vision loss has been steadily increasing, seriously compromising both the physical and mental health of patients and imposing a considerable economic burden on society. Conventional therapies currently available are largely unable to repair or regenerate the damaged retina. This review summarizes the recent advances and challenges in stem cell therapy for retinal degenerative diseases.

Abstract:
Objective To investigate whether Carthamus tinctorius L.-derived nanovesicles (CDNVs) can alleviate angiotensin II (ANGII)-induced cardiomyocyte (H9C2) hypertrophy by inhibiting the NF-κB signaling pathway and to elucidate the underlying molecular mechanisms. Methods CDNVs were isolated and purified by ultracentrifugation combined with sucrose density gradient centrifugation, and their morphology and particle size distribution were identified by transmission electron microscopy and nanoparticle tracking analysis. Rat H9C2 cardiomyocytes were cultured in vitro and randomly divided into the normal control group (NC), ANGII (1.0 μmol/L) group, ANGII+CDNVs group, and ANGII+CDNVs+NF-κB activator 1 group. After 24 h of intervention, cell viability was detected by the CCK-8 assay, the expression of myocardial hypertrophy-related genes (ANP, BNP) was determined by qPCR, the relative cell area was measured by phalloidin staining, the expression of hypertrophy marker proteins and NF-κB pathway proteins (P-p65/p65, P-IκBα/IκBα) was examined by Western blot, and nuclear translocation of p65 was observed by immunofluorescence. Subsequently, NF-κB signaling pathway agonist activator 1 was added, and the expression of myocardial hypertrophy marker proteins and NF-κB pathway proteins was detected. Results CDNVs exhibited a typical round-like vesicle structure with an average particle size of 140.4 nm, which was consistent with the morphological characteristics of exosomes. ANGII at concentrations of 0.1–1.0 μmol/L had no significant effect on cardiomyocyte viability. Compared with the NC group, ANGII significantly upregulated the gene expression of ANP and BNP, increased the relative surface area of cardiomyocytes, elevated the levels of myocardial hypertrophy marker proteins and NF_κB pathway proteins, and promoted the nuclear translocation of p65 (P<0.05). Following treatment with CDNVs, all the above indicators were markedly reduced in comparison with the ANGII group (all P<0.05). However, all these experimental results were reversed after the addition of an NF_κB agonist. Conclusion CDNVs alleviate ANGII-induced H9C2 cell hypertrophy by inhibiting the NF-κB signaling pathway.

Abstract:
Objective To explore the mechanism by which Y-box binding protein 1 (YB1) mediates the expression of multidrug resistance gene 1 (MDR1) and regulates cisplatin-induced resistance in intrahepatic cholangiocarcinoma (ICC). Methods After treating human ICC HCCC-9810 cells with different concentrations of cisplatin, Western blot and immunofluorescence techniques were used to detect the expression of YB1 and phosphorylated YB1 (pYB1) and changes in their cellular localization. In addition, qPCR and Western blot were used to analyze the expression changes of drug resistance-related proteins MDR1, multidrug resistance-associated protein 2 (MRP2), and major vault protein (MVP). Chromatin immunoprecipitation (ChIP) assay was performed to examine the binding of YB1 to the MDR1 promoter in HCCC-9810 cells with or without cisplatin treatment. Additionally, bisulfite sequencing (BS-seq) was used to determine the methylation status of cytosine-phosphate-guanine (CpG) islands within the MDR1 promoter in HCCC-9810 cells, before and after cisplatin treatment. Results After HCCC-9810 cells were treated with cisplatin, the total amount of intracellular YB1 protein did not change, but the level of pYB1 gradually increased, accompanied by the nuclear migration of YB1 and pYB1 protein. After HCCC-9810 cells were induced with cisplatin, the expression of MDR1 significantly increased, and the drug resistance of the cells also increased. If the expression of MDR1 was inhibited beforehand, the increase in drug resistance of the cells can be reversed. Similarly, after knocking down the expression of YB1 in HCCC-9810 cells, the nuclear migration of YB1 and pYB1 significantly decreased, and the expression of MDR1 significantly decreased, resulting in a significant increase in sensitivity to cisplatin chemotherapy. When HCCC-9810 cells were not stimulated by external factors, the CpG island in the promoter region of MDR1 showed a high methylation state, and multiple transcription factors, including YB1, could not bind to it. When HCCC-9810 cells were treated with cisplatin, the CpG island in the promoter region of MDR1 underwent demethylation, and the binding force between YB1 and the MDR1 promoter in the nucleus increased, activating MDR1 expression. Conclusion YB1 is highly expressed in the cytoplasm of ICC cells. Cisplatin chemotherapy induces nuclear translocation of YB1, accompanied by demethylation of the CpG island in the MDR1 promoter. Nuclear YB1 binds to the MDR1 promoter and activates transcription, ultimately leading to chemotherapy resistance.

Abstract:
Objective To investigate the toxic effects of monoethylhexyl phthalate (MEHP), a metabolite of di(2-ethylhexyl) phthalate (DEHP), on normal hepatocytes AML12 and human hepatocellular carcinoma cells HepG2, and preliminarily explore its mechanisms. Methods AML12 cells were cultured in vitro and divided into a control group and a MEHP model group, with the MEHP model group further subdivided into three concentrations (200, 400, and 800 μmol/L). The expression levels of cell morphology, cell viability, apoptosis, reactive oxygen species (ROS), superoxide dismutase (SOD), and mRNA of apoptosis-related factors were detected using methods such as RT-qPCR and Western blot, with cellular morphology observed accordingly. Results As MEHP concentration increased, cell viability was significantly inhibited, with a half maximal inhibitory concentration (IC50) of approximately 393 μmol/L. Compared to the control group, the model group exhibited significantly elevated ROS levels (P<0.001), reduced SOD activity, and a marked increase in MDA expression (P<0.001). Additionally, the expression of the anti-apoptotic factor bcl-2 decreased, while pro-apoptotic factors (bax, caspase3, etc.) increased (P<0.001), along with significantly elevated levels of inflammatory factors (IL-1, IL-6, TNF-α). Conclusion MEHP-induced liver cell damage is associated with apoptosis and oxidative stress, which have important implications for studying the mechanisms of MEHP-mediated liver injury.

Abstract:
Objective To investigate the therapeutic effect of proanthocyanidins (PC) on rats with type 2 diabetes mellitus (T2DM) complicated with cognitive impairment (CI) and to explore its potential mechanism by inhibiting the NOD-like receptor family pyrin domain-containing protein 3 (NLRP3)/cysteine-aspartic acid protease-1 (Caspase-1) pathway. Methods A T2DM rat model was established by high-fat diet combined with intraperitoneal streptozotocin (STZ) injection. Thirty-six diabetic rats complicated with CI were screened by Morris water maze test and randomly divided into model group, PC low-dose (50 mg/kg) group, PC medium-dose (100 mg/kg) group, and PC high-dose (200 mg/kg) group. Another 9 normal rats served as the normal control group. Each group was administered with the corresponding doses of PC or an equal volume of distilled water by gavage for 8 weeks. After the intervention, spatial learning and memory abilities were evaluated using the Morris water maze test. Fasting blood glucose, body weight, and organ coefficients were measured. Pathological changes in the hippocampus were observed by hematoxylin-eosin (HE) staining. Reactive oxygen species (ROS) levels, as well as levels of inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in hippocampal tissue, were detected by enzyme-linked immunosorbent assay (ELISA). The protein expression levels of NLRP3, Caspase-1, IL-1β, and IL-18 in the hippocampus were determined by Western blot. Results Compared with the normal control group, the model group exhibited significantly increased blood glucose and decreased body weight (P<0.05). In the Morris water maze test, the model group showed prolonged escape latency, fewer platform crossings, and lower percentages of time and path length in the target quadrant (P<0.05). Disordered arrangement and pyknotic nuclei were observed in hippocampal neurons. ROS and the levels of TNF-α and IL-1β were elevated, and the protein expression levels of NLRP3, Caspase-1, IL-1β and IL-18 were upregulated (P<0.05). Compared with the model group, all PC-treated groups, especially the high-dose group, presented dose-dependent improvements in the above indicators, including reduced blood glucose (P<0.05), improved spatial memory performance in the Morris water maze (P<0.01), alleviated hippocampal pathological damage, decreased ROS and inflammatory cytokine levels (P<0.05), and significantly downregulated expression of NLRP3/Caspase-1 pathway-related proteins (P<0.05). Conclusion Proanthocyanidins can ameliorate glycolipid metabolic disorder, visceral organ injury as well as cognitive dysfunction in T2DM rats. The underlying mechanism may be associated with reduced hippocampal ROS levels, inhibition of the NLRP3/Caspase-1 signaling pathway, and attenuated neuroinflammation in the hippocampus, with the optimal effect observed at a dose of 200 mg/kg.

Abstract:
Helicobacter pylori (H. pylori) infection is not only associated with gastrointestinal diseases, but also potentially links with neuropsychiatric disorders revealed by recent studies. Through a systematic review of the existing literature, this review summarizes epidemiological evidence for the association between H. pylori infection and diseases such as Parkinson’s disease (PD), Alzheimer’s disease (AD), multiple sclerosis (MS), migraine, depression, and anxiety disorders, and further explores potential mechanisms by which it affects central nervous system function, including neural pathways mediated by the enteric nervous system and the vagus nerve, immune pathways mediated by inflammatory cytokines, and endocrine pathways mediated by gut microbial metabolites. Additionally, the current challenges facing H. pylori eradication therapy and the prospects for emerging adjuvant strategies such as probiotics and fecal microbiota transplantation are discussed. Finally, future research directions are outlined, emphasizing the need for prospective cohort studies based on standardized diagnostic methods and further investigation using animal models to elucidate the pathophysiological mechanisms, with the aim of providing new perspectives for the prevention, treatment, and early intervention of related neuropsychiatric disorders.