Oxidative stress plays a key role in physiological and pathological processes occurring in the skin, leading to damage to lipids, proteins, and other cellular structures. Reliable markers of oxidative changes in the stratum corneum include lipid peroxidation products such as malondialdehyde and 4-hydroxynonenal, as well as carbonylated proteins. In recent years, particular attention has been paid to obtaining biological material for research using non-invasive methods that do not disrupt tissue integrity, allowing for an accurate evaluation of the superficial epidermal oxidative status under physiological conditions and after exposure to environmental factors. This review summarizes the properties of key oxidation products, their biological significance, and the possibilities for their detection using the tape stripping technique, with particular emphasis on its technical and analytical limitations.

Nephrogenic diabetes insipidus (NDI) is a rare disease characterized by the inability of the kidney to respond to vasopressin and concentrate urine. Hereditary NDI is primarily caused by mutations in the arginine vasopressin receptor-2 (AVPR2) gene, which enables the response to antidiuretic hormone. Mutations may lead to misfolding of the mutant AVPR2 protein or its retention within the Endoplasmic reticulum of the cell. Therefore, patients cannot respond to antidiuretic hormone and concentrate urine. Consequently, in untreated patients severe dehydration and hypernatremia may occur depending on the severity of the disease. Low-sodium and protein diet, using thiazide diuretics, and amiloride treatment are the conventional and standard treatments for NDI. Besides, many in vitro functional analysis studies have been focused on pharmacological chaperones for restoring function of NDI-causing AVPR2 mutants. Vaptans, which are known as vasopressin receptor antagonists and also pharmacological chaperones, are among the current and controversial topics for developing new treatment strategies for NDI. This study aims to review the in vitro studies on vaptans which are targeted to restore expression and function of mutant AVPR2s that cause NDI and highlight the importance of their effectiveness according to the type of mutation.

Apelin receptor (APJ) is a G protein-coupled receptor whose endogenous ligands include Apelin and Elabela (ELA), forming the Apelin/APJ and ELA/APJ signaling axes. Recent studies have shown that Apelin and ELA play a critical regulatory role in the development of hypertension through mechanisms such as modulating vascular tone, exerting anti-inflammatory and antioxidant effects, and inhibiting the renin-angiotensin-aldosterone system (RAAS). This article systematically reviews the mechanisms of the Apelin/APJ and ELA/APJ systems in hypertension, focusing on their effects on vascular endothelial function, smooth muscle proliferation, RAAS regulation, oxidative stress, and inflammatory responses. It further explores their potential as novel therapeutic targets for hypertension, aiming to provide a theoretical basis for understanding the pathogenesis of hypertension and for the development of new antihypertensive drugs.

Mitogen-activated protein kinases (MAPKs) participate in signaling pathways triggered by diverse stimuli, including stress, growth factors, and autoantibodies. Activated MAPKs play essential roles in multiple cellular processes, such as cell proliferation, apoptosis, differentiation, and immune and stress responses. Dual-specificity phosphatases (DUSPs) belong to the family of protein tyrosine phosphatases and use MAPKs as the main substrates. DUSPs exhibit physiological and pathological activities by affecting cell growth, metastasis, and death. Increasing evidence has revealed that DUSPs play essential roles in tumor initiation, progression, and therapeutic resistance. As a vital member of DUSPs, DUSP4 has potent regulatory functions in tumors by mediating proliferation, growth, metastasis, autophagy, apoptosis, and therapeutic sensitivity. Recently, studies have suggested that DUSP4 is significantly altered in colorectal cancer (CRC) and is involved in its development. In this study, we will review the expression characteristics of DUSP4 in patients with CRC and will also summarize the associated cellular functions and molecular mechanisms. We hope this study provides a reference for developing novel therapeutic strategies against CRC.

Background: Spinal cord injury (SCI) can induce secondary damage, such as inflammation, oxidative stress, and neuronal cell death. These factors impede recovery. This study evaluated the therapeutic potential of tegoprazan (TEGO), a drug known to act via a potassium-competitive acid blocker, in a SCI rat model.

Methods: We conducted SCI in female rats, and TEGO was administered intraperitoneally. Methylprednisolone (MP) was used as a positive control. TEGO was dissolved in dimethyl sulfoxide (DMSO) and sonicated before injection. Toxicity was assessed by body weight monitoring, and motor recovery was assessed by behavioral tests. We performed immunofluorescence and qRT-PCR to investigate inflammatory markers. Western blot analyses were evaluated for mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signaling pathways.

Results: TEGO treatment had no systemic toxicity and improved motor recovery. TEGO (p < 0.05) and MP (p < 0.01) decreased inducible nitric oxide synthase (iNOS) expression. CD206 expression was significantly increased in the MP group (p < 0.01) compared with the Vehicle group, whereas the TEGO group showed a slight increase compared with the Vehicle group. Furthermore, TEGO inhibited MAPK/NF-κB signaling, attenuating pro-inflammatory cytokine production.

Conclusions: TEGO, a representative potassium-competitive acid blocker (P-CABs), suppressed pro-inflammatory gene expression and might be applied as a novel treatment for SCI.

Background: Osteoporosis is a systemic skeletal disease with a multifactorial pathogenesis, and microgravity contributes significantly to its progression. Despite the critical role of epigenetic modifications, particularly DNA methylation, in cellular function, their involvement in microgravity-induced osteoporosis remains unclear. This study investigates the impact of DNA methylation on osteoblast phenotypes under microgravity, offering new insights into the epigenetic regulation of bone homeostasis.

Methods: Reduced representation bisulfite sequencing (RRBS), methylation-specific PCR (MSP), bisulfite sequencing PCR (BSP), and quantitative PCR (qPCR) were used to analyze the DNA methylation and transcriptional profiles of MC3T3-E1 cells under microgravity. Differentially methylated regions (DMRs) in the Ddit3 promoter and its expression were assessed. Functional assays, including Western blotting (WB), flow cytometry, and CCK-8, were performed to evaluate the impact of Dnmt3a overexpression or Ddit3 inhibition on osteoblast function. In vivo, a tail suspension (TS) mouse model was used to investigate bone phenotypic changes after Ddit3 inhibition. The role of the endoplasmic reticulum (ER) stress pathway in mediating Ddit3's effects was examined using the ER stress inhibitor 4-phenylbutyric acid (4PBA).

Results: Microgravity exposure markedly induced apoptosis in osteoblasts, with the apoptotic rate increasing from 3.83% in the control group to 13.44% in the microgravity group (p < 0.05). Pro-apoptotic proteins Bax and cleaved caspase-3 increased by 30% (p < 0.05) and 66% (p < 0.05), respectively, whereas the anti-apoptotic protein Bcl-2 decreased by 33% (p < 0.05). Reduced representation bisulfite sequencing (RRBS) revealed a significant global decrease in DNA methylation under microgravity. RT-qPCR analysis demonstrated a 48% reduction in Dnmt3a expression (p < 0.05), accompanied by hypomethylation of the Ddit3 promoter and a 2.36-fold elevation of Ddit3 mRNA (p < 0.05). BSP further confirmed a 62% (p < 0.05) decrease in Ddit3 promoter methylation, while MSP showed a 45% reduction (p < 0.05). Functional assays indicated that Dnmt3a overexpression elevated cell viability by 31% (p < 0.05) and suppressed apoptosis, reducing Bax expression by 35% (p < 0.05) and increasing Bcl-2 expression by 103% (p < 0.05) compared with the microgravity group. Moreover, the regulatory effects of Dnmt3a were Ddit3-dependent; inhibition of Ddit3 under microgravity reduced the apoptotic rate by 58% (p < 0.05) relative to Dnmt3a-inhibited cells. Microgravity also strongly activated the ER stress pathway, upregulating p-PERK, p-IRE1α, and XBP1s by 3.2-, 2.1-, and 3.8-fold (p < 0.05), respectively. Under microgravity, Ddit3 inhibition alleviated ER stress, decreasing p-PERK, p-IRE1α, and XBP1s by 57% (p < 0.05), 48% (p < 0.05), and 56% (p < 0.05), respectively. Furthermore, treatment with the ER stress inhibitor 4PBA under microgravity reduced the ER stress markers ATF4 and XBP1s by 48% (p < 0.05) and 49% (p < 0.05), respectively. In vivo studies using a TS mouse model confirmed that Ddit3 inhibition attenuated the osteoporosis-like phenotype, improving bone mass and microarchitectural integrity.

Conclusion: Under microgravity, Dnmt3a downregulation induces Ddit3 promoter hypomethylation, triggering ER stress and osteoblast apoptosis, thereby promoting osteoporosis. These findings identify the Dnmt3a–Ddit3 axis as a key epigenetic regulator of osteoblast dysfunction in microgravity and a promising therapeutic target for microgravity-induced osteoporosis.

Background: Type 2 diabetes mellitus (T2DM) is driven by progressive β-cell failure, wherein endoplasmic reticulum (ER) stress activates the Protein kinase R-Like ER kinase (PERK)-activating transcription factor 4 (ATF4) signaling cascade, promoting ferroptosis and accelerating β-cell dysfunction. This study investigated whether STC-1–derived exosomes (EXO) protect β-cells by modulating this pathway.

Methods: A T2DM mouse model was induced via high-fat diet (HFD) feeding and streptozotocin (STZ) administration. Exosomes were extracted from STC-1 cell culture supernatants, followed by both in vitro cellular experiments and in vivo animal studies. MIN6 cells cultured under high-glucose (HG) conditions were treated with graded concentrations of STC-1-EXO or the ferroptosis inhibitor Ferrostatin-1. Cell viability was quantified using cell Counting Kit-8 (CCK-8) assay. Western blotting (WB) assessed expression of ferroptosis markers acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4). Ferroptotic activity was evaluated by measuring intracellular total iron concentration, malondialdehyde (MDA) content, and glutathione (GSH) levels. In vivo, T2DM mice received STC-1-EXO treatment, followed by assessment of random blood glucose, glucose tolerance, insulin sensitivity, and pancreatic histopathology with concomitant protein expression analysis.

Results: In vitro, STC-1-EXO dose-dependently reversed HG-induced MIN6 cell viability loss (p < 0.05). Relative to the high-glucose control group, STC-1-EXO treatment significantly attenuated intracellular iron accumulation and MDA production, while restoring GSH levels and reducing lipid peroxidation (p < 0.05). WB results revealed that compared to the HG group, STC-1-EXO treatment significantly downregulated the expression of ER stress markers glucose-regulated protein 78 (GRP78), phosphorylated-PERK (p-PERK), phosphorylated eukaryotic translation initiation factor 2α (p-eIF2α), and ATF4, while simultaneously reducing the pro-ferroptotic protein ACSL4 expression and increasing the anti-ferroptotic protein GPX4 expression (p < 0.05). In vivo experiments showed that compared to untreated T2DM mice, STC-1-EXO-treated mice exhibited significantly decreased random blood glucose levels and markedly improved glucose tolerance and insulin sensitivity (p < 0.05). Hematoxylin and Eosin (H&E) staining revealed increased islet area and enhanced β-cell numbers in STC-1-EXO-treated mice. Furthermore, compared to untreated T2DM mice, STC-1-EXO treatment significantly reduced the expression of GRP78, p-PERK, p-eIF2α, and ATF4 in pancreatic tissue, decreased ACSL4 expression while increasing GPX4 expression, and resulted in significantly decreased MDA levels and elevated GSH levels in pancreatic tissue (p < 0.05).

Conclusion: STC-1-EXO effectively ameliorates ER stress and ferroptosis in pancreatic β-cells through inhibition of the PERK-ATF4 signaling pathway, thereby improving blood glucose levels and insulin secretory function in T2DM mice. This finding provides a novel theoretical foundation and potential therapeutic strategies for exosome-based diabetes treatment.

Background: With the advancement of technology, fully automated immunohistochemistry staining systems have been widely adopted in clinical pathological diagnosis due to their standardized operation and high reproducibility. Nevertheless, the specificity and accuracy of staining outcomes are critically dependent on the concentration of the primary antibody. Therefore, the systematic optimization of primary antibody concentration to improve immunohistochemistry staining quality remains a key issue to be addressed. This study aims to optimize the primary antibody concentration for the BOND-III platform to establish standardized protocols and enhance the reliability of immunohistochemistry staining results.

Methods: This study was performed using pathological specimens confirmed positive for a panel of biomarkers. These included the nuclear markers estrogen receptor (ER) and Ki-67, the cytoplasmic markers p16 and cytokeratin 5/6 (CK5/6), and the membranous markers human epidermal growth factor receptor 2 (HER2) and E-cadherin. Immunohistochemical staining was performed using the BOND-III platform to comprehensively assess the impact of a systematic gradient of antibody dilutions on staining quality. The resulting stains were evaluated by comparing the intensity, contrast, and degree of non-specific background across the different dilution levels. Staining scores were statistically compared among dilution groups. Differences in the rate of optimal staining were analyzed using Fisher's exact test.

Results: On the BOND-III platform, the optimal dilution ratio varied significantly for different antibodies. The ER, Ki-67, CK5/6, HER2, and E-cadherin antibodies achieved optimal staining at a 1:2 dilution, characterized by clear positive cell expression, clean background, absence of non-specific staining, and high contrast. Consistently, these five antibodies showed the highest immunohistochemical staining scores at a 1:2 dilution among all tested concentrations, with statistically significant differences compared to the neat group. In contrast, the p16 antibody demonstrated optimal staining at a 1:5 dilution ratio, showing well-defined positive cell localization and clear contrast, which was significantly superior to the neat antibody and all other dilution groups.

Conclusion: Our findings demonstrate that the optimal primary antibody dilution ratio for the BOND-III platform is antibody-dependent. Specifically, antibodies that are often used undiluted on other platforms may require further dilution on the BOND-III, with an optimal range typically falling between 1:2 and 1:5. This optimization not only improves staining quality by enhancing staining specificity and reducing background interference, but also reduces the reagent costs for routine clinical practice.

Background: Acute kidney injury (AKI) is clinically characterized by a rapid deterioration of renal function and is often associated with inflammation and oxidative stress. This study elucidated protective roles and underlying mechanisms of tight junction (TJ) proteins in AKI, with particular emphasis on the involvement of the TYRO protein tyrosine kinase binding protein (TYROBP) (also known as DNAX-associated protein 12 [DAP12])/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway.

Methods: After establishing a cisplatin-induced AKI model, TJ protein levels, inflammation and oxidative stress were evaluated. ZO-1 (TJP1)-related genes were identified using Weighted Gene Co-Expression Network Analysis (WGCNA) and Protein-Protein Interaction (PPI) network analysis. An HK-2 cell injury model was established using cisplatin, followed by assessment of the impact of DAP12 silencing on TJ proteins, inflammation, oxidative stress, and PI3K/AKT signaling pathway. Renal function, inflammation (tumor necrosis factor-α [TNF-α] and interleukin-1β [IL-1β]), oxidative stress (reactive oxygen species [ROS] and malondialdehyde [MDA]), and TJ protein levels were determined through histopathology, Western blotting and enzyme-linked immunosorbent assay (ELISA).

Results: Cisplatin-induced AKI resulted in downregulated TJ proteins (ZO-1, Occludin, Claudin-1), elevated inflammation and oxidative stress, and activated DAP12/PI3K/AKT (p < 0.05). Among the three TJ proteins, only TJP1 level was diminished in kidney injury-related diseases. Then, through WGCNA and PPI analysis, DAP12 was identified as a key gene negatively correlated with TJP1. DAP12 silencing in the HK-2 cell injury model attenuated cellular damage, restored TJ protein expression, and reduced inflammation and oxidative stress, concomitantly suppressing the activation of the PI3K/AKT pathway (p < 0.01).

Conclusion: Targeting the DAP12/PI3K/AKT pathway and TJ proteins presents therapeutic potential for AKI, and the upstream and downstream mechanisms of DAP12 in kidney injury warrant further investigation.

Background: Isoproterenol (ISO) induces oxidative and apoptotic damage in cardiomyocytes. This study aimed to investigate whether hesperidin (HES) confers a protective effect against ISO-induced injury in H9c2 cardiomyocytes and to determine whether this effect modulates the Protein Kinase B (Akt)/Glycogen Synthase Kinase-3 Beta (GSK3β) signaling pathway.

Methods: H9c2 rat embryonic cardiomyocytes were treated with ISO to establish the optimal injury concentration and with varying HES doses (5, 10, 20 μM) to define a safe and effective range (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT assay). The cells were divided into five groups: Control, ISO alone, and ISO plus HES at each concentration. The viability and proliferative capacity of the cells were assessed using MTT and 5-Ethynyl-2′-deoxyuridine (EdU) assays. Markers of injury and oxidative stress—Lactate dehydrogenase (LDH) release, glutathione (GSH), catalase (CAT), and malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity, and intracellular reactive oxygen species (ROS)—were measured. Apoptosis and mitochondrial function were assessed using Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide (JC‑1) dye for membrane potential, and Western blotting for B-cell lymphoma 2 (Bcl‑2), Bcl-2-associated X protein (Bax), Cysteine-aspartic acid protease-3 (caspase‑3), PTEN-induced putative kinase 1 (PINK1), Parkin and cytochrome C. Akt/GSK3β pathway activation was analyzed using quantitative real-time polymerase chain reaction (qRT-PCR) and phospho‑specific Western blotting. The role of Akt was confirmed by co-treatment with the Akt-specific inhibitor VIII.

Results: Compared to ISO alone, HES dose-dependently enhanced cell viability and proliferation, significantly reduced LDH release, MDA, and ROS levels, while increasing GSH and CAT levels, as well as SOD activity (p < 0.05). Results from the TUNEL assay and JC-1 staining demonstrated that treatment with HES markedly reduced the proportion of apoptotic cells and effectively maintained the integrity of the mitochondrial membrane potential (p < 0.05). Western blot analysis showed that HES treatment upregulated the levels of anti-apoptotic protein Bcl-2 and mitochondrial quality control proteins PINK1 and Parkin, while downregulating the levels of pro-apoptotic proteins Bax, caspase-3, and cytochrome C (p < 0.05). Treatment with HES notably enhanced the phosphorylation levels of both Akt and GSK3β proteins (p < 0.05), indicating activation of this signaling cascade. These protective effects were abolished in the presence of Akt inhibitor VIII, confirming reliance on Akt/GSK3β signaling.

Conclusion: Hesperidin exerts significant cardioprotective effects by alleviating ISO-induced oxidative stress, mitochondrial impairment, and apoptotic injury in H9c2 cardiomyocytes, an effect largely attributed to its activation of the Akt/GSK3β signaling pathway, thereby highlighting its therapeutic potential in cardiac protection.

Background: Therapy-related myeloid neoplasms (MN-pCT) are serious long-term complications affecting cancer survivors. This study aimed to determine the incidence, characterize the clinical and genetic features, and assess outcomes along with prognostic factors in a cohort of Chinese patients with MN-pCT.

Methods: We conducted a retrospective analysis of 460 patients diagnosed with myelodysplastic syndromes (MDS) or acute myeloid leukemia (AML) at our institution between 2008 and 2022. Among them, 31 patients met the diagnostic criteria for MN-pCT. The clinical parameters, prior treatment history, cytogenetic profiles, and mutational status were reviewed. Overall survival (OS) and disease-free survival (DFS) were calculated, and prognostic factors were identified using univariate and multivariate analyses.

Results: The incidence of MN-pCT was 6.7%, with significantly poorer median OS than primary myeloid neoplasms (15 months for AML-pCT vs. 30 months for primary AML; 7 months for MDS-pCT vs. 47.5 months for primary MDS). Breast cancer was the most common prior malignancy. Univariate analysis identified advanced age (p < 0.018), elevated lactate dehydrogenase (LDH) levels (p < 0.006), higher bone marrow blast percentage (p < 0.042), presence of TP53 mutations (p < 0.018), and complex karyotype(p < 0.015) as significant adverse prognostic factors. A novel prognostic scoring model incorporating age, TP53 mutation status, and complex karyotype effectively stratified patients into low- and high-risk groups with markedly different OS (p < 0.0305) and DFS (p < 0.0453) outcomes. Additionally, allogeneic stem cell transplantation (allo-SCT) was associated with improved survival, including in patients with high-risk features such as TP53 mutations.

Conclusions: MN-pCT is associated with poor prognosis and a high frequency of adverse genetic abnormalities. The newly developed prognostic scoring system offers a practical tool for risk stratification. Allo-SCT represents a critical and potentially curative treatment option, highlighting the need for innovative approaches to expand transplant accessibility for more affected patients.

Background: Keloid disorder manifests as the hyperproliferation of fibroblasts and resultant excessive scar formation, and the upregulation of the MyoD family inhibitor (MDFI) has been identified in keloids. Bioinformatics predictions further suggested an enrichment of sterol regulatory element binding transcription factor 2 (SREBF2) at the MDFI enhancer site. This study aimed to elucidate the specific roles and the potential regulatory axis of SREBF2 and MDFI in the proliferation of keloid fibroblasts (KFs).

Methods: The location of MDFI enhancer was predicted through bioinformatics. Chromatin immunoprecipitation followed by polymerase chain reaction was subsequently performed to quantify the enrichment of histone H3 lysine 27 acetylation (H3K27Ac) and SREBF2 at the MDFI enhancer in KFs derived from keloid dermis or normal skin fibroblasts (NFs) collected from adjacent healthy skin tissues. Following transfection as appropriate, proliferation, migration and invasion of KFs were determined through Ki67 staining, wound healing assay and Transwell assay, respectively. Western blot or quantitative real-time reverse transcription PCR analyses were applied to test the expression levels of MDFI, SREBF2, or apoptosis-related proteins (BCL2 associated X (Bax), BCL2 apoptosis regulator (Bcl-2)).

Results: MDFI and SREBF2 expression levels were upregulated in keloid dermis (p < 0.05). SREBF2 was enriched in the enhancer region of MDFI, and its silencing suppressed MDFI expression in KFs (p < 0.05). MDFI overexpression promoted proliferation, migration and invasion, elevated Bcl-2 expression in KFs, but suppressed Bax expression, whereas MDFI silencing did conversely (p < 0.05). SREBF2 silencing repressed proliferation, migration and invasion, diminished Bcl-2 expression in KFs, but augmented Bax expression, which was counteracted by overexpressed MDFI (p < 0.05).

Conclusion: This study demonstrates that SREBF2 promotes KF proliferation, migration and invasion, and inhibits apoptosis by activating MDFI through binding to MDFI enhancer transcription. The identified SREBF2/MDFI axis presents a novel potential therapeutic target for mitigating keloid progression.

Background: Alzheimer's disease (AD) is characterized by neuroinflammation, amyloid-β (Aβ) accumulation, and abnormal microtubule-associated protein tau (Tau) phosphorylation. This study evaluates the therapeutic potential of pterostilbene (PTS), a natural compound with anti-inflammatory activity, in an Aβ1-42-induced AD mouse model.

Methods: AD mice were established by intraperitoneal injection of aggregated Aβ1-42 and were subsequently treated with PTS, donepezil, or PTS combined with a β-site amyloid precursor protein–cleaving enzyme 1 (BACE1) upregulating agent. Cognitive performance was assessed through behavioral tests, and AD-related pathology was evaluated using Enzyme-linked immunosorbent assay (ELISA), immunofluorescence, reverse transcription quantitative polymerase chain reaction (RT-qPCR), and Western blotting.

Results: Compared with AD mice, PTS treatment significantly improved cognitive performance in behavioral tests (p < 0.01), reduced hippocampal Aβ immunoreactive deposition and decreased soluble and insoluble Aβ40/42 levels (p < 0.01), and markedly inhibited Tau phosphorylation at Ser396 and Thr231 (p < 0.01). PTS also significantly suppressed neuroinflammatory responses, as evidenced by reduced levels of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6), together with downregulation of microglia- and astrocyte-associated proteins (p < 0.01). At the molecular level, PTS was associated with reduced expression of BACE1, Clathrin, and Nicastrin as well as inhibition of the nuclear factor kappa B (NF-κB) pathway, as reflected by decreases in NF-κB p65 subunit (p65) and phosphorylated inhibitor of nuclear factor kappa B alpha (p-IκBα) levels, along with restoration of IκBα expression (p < 0.01). Co-treatment with the BACE1 upregulation–associated agent partially attenuated the protective effects of PTS.

Conclusion: PTS exerts significant neuroprotective effects in an Aβ1-42-induced AD mouse model by alleviating cognitive impairment, amyloidogenic pathology, neuroinflammation, and Tau hyperphosphorylation, potentially through modulation of BACE1-related processes and NF-κB signaling. PTS may represent a promising multi-target therapeutic candidate for AD.

Background: Renal ischemia-reperfusion injury (RIRI) is characterized by increased reactive oxygen species (ROS) and mitochondrial damage. Sirtuin 3 (SIRT3) promotes the deacetylation of isocitrate dehydrogenase 2 (IDH2) at lysine 413 (K413), thereby maintaining mitochondrial homeostasis. However, the roles of SIRT3 and IDH2 in RIRI remain unclear.

Methods: RIRI mouse models and hypoxia-reoxygenation (H/R)-exposed primary renal tubular epithelial cells (RTECs) were established, combined with adeno-associated virus and plasmid. Biochemical reagent kits, ELISA kits, hematoxylin and eosin staining, Transmission electron microscope, flow cytometry, Cell counting kit 8, co-immunoprecipitation and western blot assay were used. The role of SIRT3 overexpression in H/R-induced RTEC injury was investigated by plasmid transfection.

Results: Deacetylation of IDH2 (K413) ameliorated renal pathological damage and suppressed kidney injury indicators, malondialdehyde levels, and nicotinamide adenine dinucleotide (NAD) phosphate⁺/NAD phosphate hydrogen ratio, while increasing superoxide dismutase activity, glutathione/glutathione disulfide ratio, NAD-IDH activity, and ATP levels (p < 0.05). It also reduced the proportion of damaged mitochondria and mitochondrial ROS. In H/R-induced RTEC cells, deacetylation of IDH2 (K413) also decreased mitochondrial function, ROS and oxidative stress levels (p < 0.05). Additionally, we confirmed an interaction between IDH2 and SIRT3, and observed that IDH2 acetylation levels significantly increased in RIRI and H/R models (p < 0.01). After deacetylation of IDH2 (K413) treatment, acetylated IDH2 (K413) ratio, the expression levels of oxidative phosphorylation enzymes (II, III, and V), cleaved caspase-3, and phosphorylated-dynamin-related protein 1 (Ser616) were decreased, while SIRT3 and phosphorylated-DRP1(Ser637) expression increased (p < 0.05). SIRT3 overexpression alleviated cell apoptosis, decreased mitochondrial ROS, and restored mitochondrial membrane potential (p < 0.01).

Conclusion: Overexpressing SIRT3 may alleviate RIRI by deacetylating IDH2, providing a theoretical basis for the pathogenesis research of RIRI.

Background: Primary hepatocellular carcinoma (HCC) is a prevalent and aggressive malignancy with a high mortality rate. Transarterial chemoembolisation (TACE) is a widely used non-surgical treatment, but post-procedural recurrence remains a major challenge. Magnetic resonance imaging (MRI) offers a detailed assessment of tumor morphology and perfusion, while serum Protein Induced by Vitamin K Absence or Antagonist-II (PIVKA-II) has emerged as a sensitive HCC-specific biomarker. This study focused on analyzing the predictive value of preoperative MRI combined with serum PIVKA-II for recurrence after TACE in primary HCC.

Methods: A total of 101 patients with primary HCC undergoing TACE between January 2018 and March 2021 were enrolled. Patients were divided into a recurrence group (n = 52) and a non-recurrence group (n = 49), followed by comparison of MRI parameters and serum PIVKA-II levels. Variables demonstrating significant differences underwent logistic regression analysis, and the diagnostic value of the parameters with significant differences was dissected using a receiver operating characteristic (ROC) curve area under the curve (AUC).

Results: Compared with the non-recurrence group, the recurrence group exhibited a higher proportion of localized protrusions and diffuse “target sign”, lower arterial phase values, portal venous phase values, and arterial phase enhancement rates, and higher PIVKA-II levels (p < 0.05). Logistic regression analysis revealed statistically significant differences in arterial phase values, portal venous phase values, and PIVKA-II levels (p < 0.05). ROC curves demonstrated AUC values of 0.872, 0.766, 0.895, and 0.939 for arterial phase values, portal venous phase values, PIVKA-II levels, and combined prediction of post-TACE recurrence in primary HCC, respectively (p < 0.05).

Conclusions: Preoperative MRI combined with serum PIVKA-II testing holds significant predictive value for post-TACE recurrence in patients with primary HCC. Low arterial- and portal venous-phase enhancement values, together with elevated serum PIVKA-II levels, were identified as independent risk factors for recurrence. The combined use of these parameters can improve the accuracy of recurrence prediction, providing a basis for individualized postoperative management and follow-up strategies to optimize patient outcomes.

Background: Gastric cancer remains a lethal malignancy, and lipid metabolic reprogramming is increasingly recognized as a key driver of its progression. However, existing lipid metabolism-related genes (LMGs) signatures lack robust external validation. In this study, we identified differentially expressed LMGs, constructed and externally validated a prognostic model, and evaluated its clinical relevance with respect to the tumor microenvironment, immune escape, and drug sensitivity, while also exploring potential therapeutic agents.

Methods: LMGs were obtained from the Gene Set Enrichment Analysis (GSEA) repository. Transcriptome data derived from The Cancer Genome Atlas (TCGA) Stomach Adenocarcinoma cohort were systematically analyzed to identify genes exhibiting significant expression differences. Functional characterization and biological pathway interpretation were subsequently performed based on the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases. Prognosis-related genes were initially screened using univariable Cox regression analysis, after which a least absolute shrinkage and selection operator approach was employed to construct a risk prediction model. The predictive capacity of the constructed model was systematically evaluated using time-to-event analyses, discrimination assessment based on Receiver Operating Characteristic (ROC) methodology, and multivariable independence testing. Furthermore, a clinically oriented nomogram was established by integrating relevant clinicopathological parameters to improve translational utility. Differences in biological pathway activity, immune-related responses, and susceptibility to chemotherapeutic agents across stratified risk categories were systematically explored using gene set enrichment analysis, the Tumor Immune Dysfunction and Exclusion algorithm, and drug response data derived from the Genomics of Drug Sensitivity in Cancer resource.

Results: A total of 179 differential expression lipid metabolism–associated genes were detected. Subsequent functional enrichment analyses revealed that these genes are primarily involved in lipid droplet organization, fatty acid and sphingolipid metabolism, and the peroxisome proliferator-activated receptor (PPAR) signaling pathway. A prognostic model was established based on eleven key genes, and patients stratified into the high-risk subgroup exhibited markedly reduced overall survival compared with the low-risk subgroup in both the training and the GSE15459 validation dataset. The prognostic model showed the area under the ROC curve (AUC) of 0.618, 0.688, and 0.734 for 1-year, 3-year, and 5-year survival intervals, respectively, with a nomogram C-index of 0.6803. Immune characterization combined with GSEA indicated that patients classified as the high-risk subgroup exhibited significant activation of pathways associated with the extracellular matrix and focal adhesion with elevated Tumor Immune Dysfunction and Exclusion (TIDE), T-cell dysfunction, and immune-exclusion scores, whereas low-risk patients showed enrichment in amino acid metabolism and DNA repair pathways, higher microsatellite instability (MSI), and distinct drug sensitivities. Analysis of chemotherapeutic responsiveness indicated that individuals classified as the high-risk subgroup exhibited increased sensitivity to 5-fluorouracil, afatinib, and docetaxel, whereas low-risk patients showed greater sensitivity to dasatinib, AZD1332, and BMS-754807.

Conclusion: We developed and validated an 11-gene lipid metabolism–based prognostic model for gastric cancer that demonstrated strong predictive performance and clinical applicability. The signature stratifies patients based on risk, reflects immune escape features and chemotherapy sensitivity, and holds potential as a tool for personalized prognosis and therapeutic decision-making.

Background: Glucocorticoid resistance in asthma is frequently associated with persistent airway inflammation and oxidative stress. Although vitamin D has been reported to exert anti-inflammatory effects and restore steroid sensitivity, the underlying signaling mechanisms remain unclear. Given that Semaphorin 7A (Sema7A) and its receptor Integrin-β1 are implicated in immune activation and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)-mediated inflammatory responses, this study aimed to elucidate how Vitamin D confers protection against bronchial inflammation and redox imbalance, emphasizing the potential regulatory role of the Sema7A/Integrin-β1/NF-κB axis.

Methods: To model allergic airway disease, we utilized a murine asthma model induced by ovalbumin (OVA) and Lipopolysaccharide (LPS)-treated human bronchial epithelial (BEAS-2B) cells. Airway inflammation and oxidative stress were evaluated using enzyme-linked immunosorbent assay (ELISA), histopathological examination, and biochemical assays. In vitro cell proliferation, cytokine secretion, and reactive oxygen species (ROS) generation were measured. The expression of Sema7A, Integrin-β1, and NF-κB-related proteins was analyzed by Western blotting and reverse transcription quantitative PCR (RT-qPCR). Functional rescue assays were conducted by overexpressing Sema7A in BEAS-2B cells.

Results: In the OVA-induced asthma mouse model, vitamin D treatment markedly reduced Th2 cytokines (IL-4, IL-5, IL-13; all p < 0.01) while restoring interferon-gamma (IFN-γ) levels (p < 0.05), accompanied by attenuated airway inflammation and oxidative stress, as evidenced by elevated superoxide dismutase (SOD) and catalase (CAT) activities (p < 0.05) and decreased malondialdehyde (MDA) content (p < 0.001). In LPS-stimulated BEAS-2B cells, vitamin D at 1 nM and 10 nM similarly suppressed tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) production, mitigated intracellular ROS accumulation, and rescued proliferative impairment (all p < 0.05). Mechanistically, Vitamin D inhibited activation of the Sema7A/Integrin-β1/NF-κB signaling cascade in both in vivo and in vitro systems, whereas Sema7A overexpression reactivated this pathway and partially diminished the anti-inflammatory effects of vitamin D.

Conclusion: Vitamin D mitigates airway inflammation and oxidative stress in allergic asthma by suppressing the Sema7A/Integrin-β1/NF-κB signaling pathway. These findings reveal a novel mechanistic insight into the anti-inflammatory action of vitamin D and highlight its therapeutic potential in allergic airway diseases.

Background: Early Growth Response 2 (EGR2) is a transcription factor implicated in inflammation and cell stress, yet its role in the pathogenesis of pediatric type 1 diabetes (T1D) remains largely unexplored. Moreover, the molecular mechanisms linking EGR2 to pancreatic β-cell dysfunction in T1D are unknown. This study aimed to elucidate the role of EGR2 in T1D progression and to explore the underlying molecular mechanisms.

Methods: The expression profile of dataset GSE9006 was analyzed using R software to assess EGR2 expression in peripheral blood mononuclear cells (PBMCs) from children with T1D. Bioinformatic analyses were further complemented by experimental validation to confirm these findings. The effects of EGR2 on MIN6 cell function were examined using Cell counting kit-8 (CCK-8) assay, 5-Ethynyl-2′-deoxyuridine (EdU) proliferation assay, flow cytometry, glucose-stimulated insulin secretion, quantitative PCR (qPCR), and immunoblotting. The regulatory influence of EGR2 on insulin receptor substrate 2 (IRS2) transcription was examined using qPCR and immunoblotting. In vivo, apoptosis was assessed by immunoblot analysis and Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, pancreatic islet injury was evaluated using hematoxylin and eosin (H&E) staining, and blood glucose levels were measured with a glucose meter.

Results: EGR2 is highly expressed in PBMC in children with T1D. In addition, EGR2 knockdown alleviated Cytomix-induced MIN6 cell dysfunction and inhibited IRS2 transcription. EGR2 knockdown ameliorated disturbances in STZ-induced blood glucose levels in mice, mitigated pancreatic islet injury in mice, and reduced apoptosis of mouse islet cells.

Conclusion: EGR2 knockdown enhances IRS2 transcription and mitigates the pathological features of T1D both in vivo and in vitro, suggesting that EGR2 may serve as a key regulator in T1D progression.

Background: Insulin-resistant polycystic ovary syndrome (IR-PCOS) is a refractory endocrine disorder with limited therapeutic options. Wogonin, a natural flavonoid compound, has shown potential therapeutic efficacy; however, its underlying mechanism remains unclear. This study aimed to explore the therapeutic effects of wogonin on IR-PCOS and the mechanistic involvement of fibronectin 1 (FN1) and phosphatidylinositol 3-kinase (PI3K)/Protein kinase B (AKT) signaling pathway.

Methods: The interaction between wogonin and its potential target was predicted using molecular docking. An IR-PCOS mouse model was treated with wogonin alone or in combination with tail-vein injection of shFN1 or short hairpin negative control (shNC) lentiviral particles. Metabolic parameters, sex hormones, ovarian morphology, and key pathway proteins were assessed using glucose/insulin tolerance tests, enzyme-linked immunosorbent assays (ELISA), histopathology, and Western blotting.

Results: Molecular docking demonstrated a stable binding between wogonin and FN1. In IR-PCOS mice, wogonin significantly reduced body weight, restored serum sex hormone imbalances—characterized by decreased testosterone, estradiol, luteinizing hormone (LH), and the LH/follicle-stimulating hormone (FSH) ratio alongside increased progesterone—and improved glucose metabolism and insulin sensitivity. It also normalized ovarian morphology, reduced cystic follicles, promoted follicular development, and inhibited granulosa cell apoptosis. Mechanistically, wogonin modulated the insulin signaling pathway and activated the FN1/PI3K/AKT pathway in ovarian tissues. Notably, these therapeutic effects of wogonin were substantially reversed by shFN1.

Conclusion: Wogonin exerts comprehensive therapeutic effects against IR-PCOS by targeting FN1 to activate the PI3K/AKT pathway, identifying FN1 as a key mechanistic target.

Background: Sepsis-associated encephalopathy (SAE) is defined by neurological impairment resulting from systemic infections. The gut microbiota has been shown to affect the brain via the gut–brain axis (GBA) in SAE. However, the underlying molecular mechanisms are unclear. This research focused on examining the potential mechanisms by which the gut microbiota and the GBA are involved in SAE by integrating multi-omics data.

Methods: An SAE model was established using 7–8-week-old male rats via lipopolysaccharide (LPS) injection combined with caecal ligation and puncture (CLP). The gut microbiota (via 16S rRNA sequencing) and metabolomes were profiled to analyze variations in gut microbiota composition and metabolite abundance in the faeces and hippocampus. Western blot and RT-qPCR analyses were applied to measure the expression of proteins or genes associated with autophagy in the hippocampus.

Results: SAE rat models presented significant differences in gut microbiota composition, characterized by reduced abundances of the genera Butyricimonas, Muribaculaceae, and Bacteroides and increased abundance of the genera Aeromonas and Family_XIII_UCG-00. Integrated analysis of untargeted metabolomics data for the faeces and metabolomics data for the hippocampus revealed that glycerophospholipid metabolism was altered in both the faeces and hippocampus in SAE rat models. Molecular analyses revealed that the activity of hippocampal autophagy pathways was reduced in SAE rat models, in contrast with rats in the non-SAE group and the control group.

Conclusion: Combining multi-omics data with molecular analysis data revealed a potential association among the gut microbiota, host glycerophospholipid metabolism and reduced neuronal autophagy in SAE model rats. The study contributes to a better understanding of the gut microbiota's involvement in SAE.

Background: Ferroptosis has emerged as a pivotal mechanism in epilepsy development. Kelch-like ECH-associated protein 1 (KEAP1), a critical regulator of the nuclear factor erythroid 2-related factor 2 (NRF2) pathway, plays a pivotal role in oxidative stress and is implicated in seizure recurrence. This study aimed to elucidate the mechanistic role of KEAP1 in kainic acid (KA)-induced ferroptosis and neuroinflammation.

Methods: HT22 hippocampal neuronal cells were stimulated with KA to establish the in vitro excitotoxicity model. Cell viability and apoptosis were detected using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively. Inflammatory responses were evaluated by detecting interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) secretion levels. Ferroptosis was evaluated by measuring Fe²⁺, glutathione (GSH), and reactive oxygen species (ROS) levels. The parkin RBR E3 ubiquitin protein ligase (PRKN)-KEAP1 and histone deacetylase 6 (HDAC6)-PRKN interactions were assessed by co-immunoprecipitation (Co-IP) experiments.

Results: KEAP1 (p = 0.0008) and HDAC6 (p = 0.0004) protein levels were upregulated in the KA-induced HT22 cells, while PRKN expression was downregulated (p = 0.0034). Knockdown of KEAP1 alleviated KA-induced neuroinflammation by reducing the secretion of IL-6 (p = 0.0021) and TNF-α (p = 0.0003). Furthermore, it suppressed ferroptosis, as demonstrated by decreased levels of Fe²⁺ (p = 0.0022) and ROS (p < 0.01), and an increased GSH content (p = 0.0057). In addition, KEAP1 silencing attenuated apoptosis (p = 0.0016) in KA-induced cells. Mechanistically, PRKN mediated KEAP1 ubiquitination and degradation (p = 0.0054); HDAC6 induced PRKN deacetylation (p < 0.001), and HDAC6 modulated KEAP1 expression via PRKN. In KA-treated HT22 cells, KEAP1 reconstitution counteracted PRKN overexpression-mediated anti-inflammatory and ferroptosis-inhibitory effects (p < 0.05), while PRKN knockdown reversed the protective effects conferred by HDAC6 silencing (p < 0.05). Additionally, PRKN regulated the NRF2/solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) pathway through KEAP1 (p < 0.05).

Conclusion: Our study uncovers a novel HDAC6-PRKN-KEAP1 cascade that critically modulates KA-induced ferroptosis and neuroinflammation in HT22 neuronal cells.

Background: Due to their unpredictable and variable nature, novel viruses pose a significant threat to human health as well as to the economy and society. Rapid detection plays a crucial role in the surveillance and management of the transmission and emergence of novel viruses. This study aims to establish a rapid, label-free surface-enhanced Raman scattering (SERS) platform for the highly sensitive and specific identification of emerging viruses.

Methods: A novel self-assembly strategy based on the synergistic effect of electrostatic adsorption and hydrogel contraction has been developed to construct highly ordered hexagonally close-packed Au nanoparticle arrays on both surfaces of a polyacrylamide composite hydrogel film. The drying and shrinking of the hydrogel film contribute to the reduction of the particle gap, facilitating the formation of “hot spots” that effectively enhance near-field coupling and significantly amplify the local electromagnetic field in its vicinity. The label-free SERS detection of viruses relies on the intrinsic Raman fingerprints of viral structural components, particularly the envelope glycoproteins and lipid membranes that differ distinctly from host cellular materials.

Results: The detection of malachite green and crystal violet on the substrate indicates high Raman optical activity. Validation with probe molecules (crystal violet, malachite green) showed low limits of detection (LOD of 10^–11 M) and high linearity (R² > 0.996). Highly linear quantitative sensitivity was observed, with a limit of detection of 1 TU/mL for both severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Spike pseudovirus and B6R Mpox pseudovirus.

Conclusion: The composite material is anticipated to serve as a novel Raman substrate, enabling the rapid identification of novel viruses and facilitating a prompt response to epidemics.

Background: Myocarditis is a rare, potentially fatal condition that particularly affects young people, with diverse causes ranging from viral infections (the most common in developed nations) to autoimmune diseases and toxins. Although acute viral myocarditis (VMC) is frequently subclinical and may resolve spontaneously, persistent inflammation causes continual myocyte damage, leading to intractable heart failure or death. Treatment options for chronic myocarditis remain inadequate. The primary purpose of this study was to investigate whether C-28 methyl ester for 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO-Me), a potent Nuclear factor erythroid 2-related factor 2 (Nrf2) activator, alleviates coxsackievirus B3 (CVB3)-induced myocardial injury in viral myocarditis, potentially through activating the Nrf2/heme oxygenase-1 (HO-1) signaling axis to exert anti-inflammatory and antioxidant effects.

Methods: Male BALB/c mice were subjected to CVB3 treatment to establish a CVB3-induced VMC mouse model, and body weight loss and survival were documented. Then, an enzyme-linked immunosorbent assay (ELISA) for measuring myocardial injury marker and inflammatory indicator contents in plasma was conducted. Echocardiography was done to detect the mouse cardiac function. To observe alterations in myocardial tissue morphology, hematoxylin-eosin (H&E) staining was conducted. Furthermore, nucleotide oligomerization domain-, leucine-rich repeat-, and pyrin domain-containing protein 3 (NLRP3), apoptotic-associated speck-like protein containing a CARD (ASC) and Caspase-1/Cleaved contents in VMC mice and H9C2 cells were measured through Western blotting (WB), immunofluorescence analysis, and quantitative real-time polymerase chain reaction (qRT-PCR). Nrf2, P-Nrf2, HO-1 and NAD(P)H quinone oxidoreductase 1 (NQO-1) levels were also examined through immunofluorescence, WB, and qRT-PCR in mice and H9C2 cells after treatment with CVB3. Moreover, for exploring how Nrf2 affected CVB3-treated VMC in vitro, Nrf2 was downregulated in H9C2 cells by small interfering RNA (siRNA) transfection.

Results: In vivo experiments revealed that bardoxolone methyl (CDDO-Me) intervention significantly improved survival, body weight loss, myocardial injury, inflammatory reactions and ventricular systolic dysfunction in CVB3-induced VMC (p < 0.05, p < 0.01, or p < 0.001 vs. CVB3 group, where applicable). Immunofluorescence, WB, and qRT-PCR analyses further revealed that the expression of inflammasome-associated proteins such as NLRP3, ASC and Caspase-1/Cleaved Caspase-1 was significantly increased in CVB3-induced VMC mice (p < 0.001), and this change was reversed by CDDO-Me intervention (p < 0.01 or p < 0.001). Notably, the downregulation of Nrf2/HO-1 pathway-related proteins in CVB3-induced VMC mice was reversed by CDDO-Me treatment (p < 0.05, p < 0.01, or p < 0.001). In vitro, we observed that CDDO-Me intervention ameliorated the CVB3-induced decrease in viability and inflammatory reactions in H9C2 cardiomyocytes (p < 0.001). Correspondingly, NLRP3, ASC and Caspase-1 were highly expressed in H9C2 cells co-incubated with CVB3 (p < 0.001), and this high expression could be reversed by CDDO-Me intervention (p < 0.05, p < 0.01, or p < 0.001), suggesting that CDDO-Me intervention could reduce inflammation levels in H9C2 cells co-incubated with CVB3. Moreover, similar to the in vivo experimental results, our in vitro study suggested that the downregulating Nrf2/HO-1 pathway-related proteins in H9C2 cells co-incubated with CVB3 was also significantly reversed by CDDO-Me intervention (p < 0.05, p < 0.01, or p < 0.001). We then transfected H9C2 cells with Nrf2-specific siRNA to silence Nrf2, as Nrf2 knockdown significantly suppressed H9C2 cell survival and enhanced the inflammatory response (p < 0.05, p < 0.005, or p < 0.001).

Conclusion: CDDO-Me reduces myocardial injury and inflammatory reactions in CVB3-induced viral myocarditis, resulting from a reduction in the NLRP3 inflammasome induced by the regulation of Nrf2/HO-1.

Background: Poly(ADP-ribose) polymerase inhibitors (PARPis) are effective adjunctive therapies for ovarian cancer (OC). However, drug resistance remains a significant clinical challenge. Current research focuses on target expression and functional changes in established resistant cell lines. Yet, a comprehensive analysis of transcriptomic alterations over time after PARPis exposure is lacking. This study aims to investigate the transcriptomic changes in OC cell lines following prolonged PARPis treatment and to identify potential resistance biomarkers.

Methods: SKOV3 and OVCAR8 cell lines were treated with Olaparib and Niraparib for 2, 3 and 6 months, and then RNA sequencing (RNA-seq) was performed. Differentially expressed genes (DEGs) were identified through “limma” analysis, following the conduction of function enrichment analysis. Survival analysis was performed using The Cancer Genome Atlas (TCGA) data to correlate DEG expression with clinical outcomes. To validate the identified key resistance genes, we modulated their expression using specific siRNAs for knockdown and overexpression vectors. We performed these experiments in ovarian cancer cells following long-term culture with PARPis. Models included SKOV3 and OVCAR8 cells, which were maintained for 6 months with either Niraparib (SKOV3_Nira_6M, OVCAR8_Nira_6M) or Olaparib (SKOV3_Ola_6M, OVCAR8_Ola_6M). We then assessed proliferation and viability to confirm the functional relevance of these genes to PARPis sensitivity in ovarian cancer.

Results: The transcriptional profiles of the same type of OC cells stimulated by Niraparib and Olaparib for 6 months were highly similar. After SKOV3 cells were treated with PARPis for 6 months, 59 DEGs showed consistent changes, while 103 DEGs were identified in OVCAR8 cells (adjusted p < 0.05 and |log₂ fold change| >1). These DEGs were enriched in biological processes such as epithelial cell differentiation (p < 0.05), estrogen receptor signaling (p < 0.05), and cell adhesion (p < 0.05), among others. Among them, we found that CPAMD8 (p < 0.05) and EGR1 were specifically upregulated in SKOV3 (adjusted p < 0.05) and OVCAR8 (adjusted p < 0.05) cells after 6 months of PARPis stimulation, and they were significantly associated with poor overall survival in OC patients. Further validation in vitro confirmed that both mRNA and protein levels of CPAMD8 and EGR1 were significantly elevated in SKOV3 and OVCAR8 cells following 6 months of maintenance culture with PARPis. Building on these findings, functional studies demonstrated that enforced overexpression of CPAMD8 or EGR1 in these PARPis-persistent cells markedly enhanced cell viability. In contrast, knockdown of CPAMD8 or EGR1 expression effectively suppressed cell viability.

Conclusion: CPAMD8 and EGR1 were identified as potential biomarkers of mild drug resistance and were associated with poorer survival.

Background: The optimal serum thyroglobulin (Tg) threshold for performing ¹⁸F-fluorodeoxyglucose positron emission tomography/computed tomography (¹⁸F-FDG PET/CT) in patients with recurrent differentiated thyroid carcinoma (DTC) and negative radioiodine scintigraphy (TENIS syndrome) remains controversial. This study aimed to systematically evaluate the relationship between Tg levels and PET/CT detection rates and to determine if a clinically meaningful threshold exists.

Methods: A systematic literature search was conducted in Web of Science, PubMed, and Cochrane Library up to January 2026 following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Studies reporting Tg-stratified PET/CT detection rates in adult patients with suspected recurrent DTC were included. The primary outcome was the per-patient detection rate, which was pooled using a random-effects model. Risk of bias was assessed using QUADAS-2.

Results: Eight studies met the inclusion criteria, with six contributing to the quantitative synthesis. The overall pooled detection rate was 0.54 (95% CI 0.29–0.78), with significant heterogeneity across studies (I² = 93.4%). A pronounced threshold effect was observed: detection rates were negligible in low-Tg strata (<5 ng/mL), increased substantially in intermediate strata (5–10 ng/mL), and were consistently high in high-Tg strata (>10 ng/mL). Specifically, the pooled detection rate was 0.71 (95% CI 0.00–1.00) for patients with Tg >10 ng/mL and increased to 0.97 (95% CI 0.90–0.99) in the subgroup with Tg ≥18 ng/mL, although this estimate was based on a limited number of studies.

Conclusion: ¹⁸F-FDG PET/CT diagnostic yield demonstrates a monotonic increase with rising serum Tg levels. A Tg threshold in the range of approximately 10–18 ng/mL appears promising for identifying structural disease, although the upper bound of this threshold warrants further validation in larger cohorts. These findings support a Tg-guided, risk-stratified approach to selecting candidates for ¹⁸F-FDG PET/CT.