Zinc transporters, including ZIP6 (SLC39A6, also known as LIV-1) and ZIP10 (SLC39A10), are reportedly involved in breast cancer progression and metastasis. We aimed to clarify the association of the expression of zinc transporters with clinicopathological features and prognosis in breast cancer. We retrospectively included 200 patients who underwent surgery for invasive ductal carcinoma of the breast (invasive diameter ≥ 10 mm) at our institution between January 1, 2010 and June 30, 2023. Immunohistochemical staining for ZIP6 and ZIP10 was performed, and their expression levels were evaluated. Survival analyses were performed by stratifying patients into four groups based on combined ZIP6 and ZIP10 expression (high/low). We found that the expression levels of ZIP6 and ZIP10 showed a statistically significant positive correlation. Low ZIP6 expression was associated with aggressive clinicopathological features, including a higher Ki-67 labeling index, higher histological grade, and larger tumor size. Although survival differences did not reach statistical significance, combined evaluation of ZIP6 and ZIP10 expression revealed distinct survival tendencies among the expression groups. These findings suggest that combined assessment of ZIP6 and ZIP10 expression may provide additional prognostic information beyond ZIP6 expression alone. Future studies with larger cohorts and longer follow-up are warranted to validate these findings.
Neutrophil extracellular traps (NETs) are web-like chromatin structures released by neutrophils that play dual roles in host defense and tissue injury. In liver diseases, NETs have emerged as key pathogenic mediators. This narrative review synthesizes current evidence on the involvement of NETs in alcohol-associated liver disease (ALD), metabolic dysfunction-associated steatotic liver disease (MASLD), viral hepatitis, hepatic ischemia–reperfusion injury (HIRI), hepatocellular carcinoma (HCC), and intrahepatic cholangiocarcinoma (iCCA). Mechanistically, NETs amplify sterile inflammation, promote hepatocyte death, activate hepatic stellate cells to drive fibrosis. NETs also promote malignant transformation, sustain an aggressive phenotype, attenuate immunotherapy responses, mediate resistance to targeted therapies, and contribute to poor prognosis in primary liver cancers. Preclinical studies targeting NET formation or degrading existing NETs have shown therapeutic benefits across these liver diseases. Despite promising results, clinical translation remains limited, with challenges including a lack of standardized biomarkers, optimal timing of intervention, and the need for liver-specific delivery. This review highlights NETs as a druggable target in liver diseases, and underscores future directions toward biomarker development and targeted therapies.
The lymphatic system in the liver has long been regarded as a passive transport route responsible for interstitial fluid drainage. However, recent research has redefined it as a “new frontier in hepatology,” playing active and multifaceted roles in liver disease progression, ascites formation, and tumor immune responses. In malignancies such as liver cancer, lymphatic vessels function as conduits for lymph node metastasis, while simultaneously serving as pathways for tumor antigens and tumor-induced inflammatory cells, potentially contributing to the modulation of the immune microenvironment. This review first outlines the latest findings regarding the lymphatic system in non-neoplastic tissues. Next, we examine recent studies investigating the impact of lymphatic vessels on cancer progression. Furthermore, we focus specifically on the hepatic lymphatic system in both non-neoplastic and cancerous states. Finally, we conclude by discussing the potential role of hepatic lymphatic vessels in regulating the tumor microenvironment in liver cancer.
Accurate prognostic stratification of endometrial carcinoma (EC) remains challenging in resource-limited settings lacking molecular sequencing. We investigated whether immunohistochemical (IHC) assessment of combined L1CAM/β-catenin expression, integrated with mismatch repair (MMR) status and p53 expression, could refine prognostic risk stratification in EC in absence of POLE sequencing. A retrospective cohort study evaluated 140 surgically staged EC cases for L1CAM, β-catenin MMR, and p53 IHC expression with clinicopathological correlation. Survival analysis was performed on 111 cases (median follow up:38 months). L1CAM and β-catenin demonstrated mutually exclusive expression patterns (27.1
Multiple myeloma (MM) is a malignant plasma cell neoplasm characterized by marked biological heterogeneity and complex metabolic reprogramming. Although dysregulated lipid metabolism has been implicated in MM, its morphological correlations and clinical significance remain unclear. The present study evaluated cytoplasmic vacuoles in peripheral blood smears and analyzed their associations with the prognosis and stage of MM. Additionally, lipid droplet formation and the expression of key cholesterol regulatory molecules among MM cell lines were examined via molecular and biochemical approaches. Intracytoplasmic vacuoles were more frequently observed at advanced Revised International Staging System stages of MM, although no significant difference in overall survival was observed. MM cell lines exhibited prominent lipid droplet accumulation and upregulated acyl-coenzyme A:cholesterol acyltransferase-1 (ACAT-1) and scavenger receptor class B type I (SR-BI) expression. The pharmacological inhibition of SR-BI suppressed MM cell proliferation and induced apoptotic cell death, whereas the inhibition of ACAT-1 had only a modest effect. Ultrastructural analysis revealed marked cytoplasmic vacuolization following SR-BI inhibition. These findings indicate that altered cholesterol metabolism is closely associated with cytoplasmic vacuolization in MM cells, suggesting that SR-BI plays a critical role in MM cell survival. Therefore, targeting the cholesterol trafficking pathways may represent a potential therapeutic strategy in MM.
Osteosarcoma has a poor prognosis and frequently metastasizes to the lungs. This study examined the role of transglutaminase 2 (TGM2), a ligand for adhesion G protein-coupled receptor 56 (GPR56), in lung metastasis of LM8 mouse osteosarcoma cells. Gβγ‑dependent ERK signaling mediated by GPR56 and the adhesion of GPR56-expressing LM8MOCK cells and GPR56‑deficient LM8LOW cells were analyzed using siRNA, western blotting, and cell staining. TGM2 and Ki67 expression in metastatic lung lesions was assessed by transplantation and immunohistochemistry. TGM2 enhanced endogenous ERK signaling in LM8MOCK cells. In the absence of TGM2, ERK signaling increased in LM8LOW cells but remained unchanged in LM8HIGH cells compared with LM8MOCK cells. In the presence of TGM2, ERK activity in LM8LOW cells resembled LM8MOCK, whereas it was decreased in LM8HIGH cells. LM8LOW cells showed reduced adhesion, whereas LM8HIGH cells did not differ from LM8MOCK cells. Proliferation did not differ between LM8LOW cells and LM8HIGH cells. TGM2 was co‑expressed in CD31 + endothelial cells, and Ki67‑positive LM8 cells were observed along bronchi. These findings suggest that GPR56 promotes lung metastasis through TGM2‑mediated adhesion to pulmonary endothelial cells and enhanced ERK‑dependent proliferative signaling.
Ulcerative colitis is a chronic inflammatory bowel disease characterized by persistent inflammation of the colon. The extensive and persistent mucosal damage associated with ulcerative colitis (UC) can contribute to carcinogenesis, thereby underscoring the significant clinical interest in identifying an in situ marker. p53-binding protein (53BP1) is a DNA damage response (DDR) molecule that localizes at sites of double-strand breaks. Herein, we investigated the in situ DDR in UC by evaluating 53BP1 immunofluorescence. Our study revealed a significant increase in abnormal 53BP1 foci, defined as three or more foci and/or foci larger than 1 µm within the nucleus, in patients with UC compared to controls. Furthermore, the presence of abnormal 53BP1 foci in UC correlated with the severity of symptoms, endoscopic gradings, serological and histopathological inflammation. Of note, 53BP1 foci were observed in the colon mucosa of patients in remission, indicating that 53BP1 is a sensitive DDR marker. In addition, large 53BP1 foci, indicative of a severe DDR, were more prevalent in patients with colitic cancer than in controls. In conclusion, our findings suggest that 53BP1 may serve as a in situ marker reflecting the extent of the DDR in UC.
This study aimed to assess the effect of a combination of osteoblasts and octacalcium phosphate collagen composite (OCP/Col) on bone regeneration. In oral and maxillofacial surgery, OCP/Col has been clinically used as artificial bone. Although OCP/Col was reported to promote osteoblast differentiation, few studies have reported on its use in combination with cell transplantation. Moreover, its effects are not fully clarified. Therefore, we evaluated the osteogenic properties of a combination of OCP/Col and osteoblasts. We found that osteoblasts cultured in OCP/Col differentiated toward osteolineage, expressing several osteoblast markers. In animal experiments, the cell transplantation group showed promotion of osteoblast differentiation and stromal formation in the tissues compared with the OCP/Col group. These findings indicate that combining osteoblasts and OCP/Col would be beneficial for bone regeneration therapy.
The APC (adenomatous polyposis coli) gene, which was first discovered as a colorectal cancer suppressor gene, is highly expressed in the nervous system. The Apc gene/Apc protein is deeply involved in brain development and morphogenesis. In the postnatal brain, Apc/Apc is involved in synaptic transmission, axon growth, and intracellular transport. Apc/Apc expressed in glial cells is involved in glial cell differentiation and neural circuit formation through glial cell functions. Reports from disease model animals and familial adenomatous polyposis (FAP) patients suggest that APC may be involved in the onset of autism, cognitive impairment, and schizophrenia.
Bisphosphonate-related osteonecrosis of the jaws (BRONJ) is a major adverse effect of bisphosphonates, yet its underlying pathogenesis remains poorly understood. Bone metabolism and remodeling relies on the interaction between osteoblasts (OBs) and osteoclasts (OCs). Sphingosine 1-phosphate (S1P), a bioactive sphingolipid metabolite, is an important mediator of OC-OB communication. In this study, we aimed to investigate the role of the S1P/S1P receptor (S1PR) axis in the development of BRONJ. A co-culture system was used to examine the interaction between OCs and OBs. Western blot and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were used to detect the expression of related proteins and messenger ribonucleic acids (mRNAs). Finally, an in vivo BRONJ mouse model was used to validate the role of S1P/S1PR axis in disease progression. In our study, we showed that zoledronate (ZOL) promoted S1P secretion from OCs and enhanced the migration of osteoclast precursor cells (OCPs) through S1PR signaling. In addition, OCs promoted the excessive osteogenic differentiation and migration of OBs via S1P/S1PR axis. Importantly, pharmacological inhibition of S1PR facilitated the recovery of BRONJ-like lesions in vivo. In conclusion, these findings indicate that the S1P/S1PR axis plays an important role in the pathogenesis of BRONJ and may represent a potential therapeutic target for its treatment.
Trimethyltin (TMT) is widely used to model hippocampal degeneration in rodents, but its effects on the kidney are poorly understood. This study investigated renal function, histology, inflammation, and Klotho expression in TMT-exposed rats. Ten male Sprague–Dawley rats (8–10 weeks) were divided into control and TMT groups (n = 5 each). TMT-treated rats received a single intraperitoneal dose (8 mg/kg). On day 28, serum urea and creatinine were measured. Kidney morphology was examined by Hematoxylin–Eosin and Periodic Acid Schiff staining. TNF-α and Klotho localization were assessed by immunohistochemistry, and Klotho and Nrf2 mRNA levels were quantified by qPCR. Body weight and renal function were similar between groups, indicating preserved kidney function. Histology revealed mild inflammatory cell infiltration in TMT-exposed kidneys and PAS staining showed mesangial expansion. TNF-α expression increased in distal tubules, while Klotho protein was elevated in cortical distal and medullary tubules, with higher semiquantitative scores than controls. Klotho and Nrf2 mRNA levels remained unchanged, suggesting post-transcriptional regulation. At 28 days post-TMT exposure, kidneys showed mild inflammation without functional impairment. Increased Klotho protein likely reflects a compensatory adaptive response to stress, indicating renal protective mechanisms following systemic TMT toxicity.
Fbxw7, a substrate recognition subunit of the SCF ubiquitin ligase complex, regulates the proteasomal degradation of multiple cancer-related and metabolic proteins. To elucidate its role in hepatic lipid homeostasis, we established Fbxw7 knockdown (FKD) Huh-7 cells and analyzed associated morphological and molecular alterations. Lipid droplets were evaluated by BODIPY staining and transmission electron microscopy, while metabolic and autophagy-related factors were examined using immunocytochemistry, Western blotting, and RT-PCR. FKD resulted in marked lipid droplet accumulation and upregulation of lipogenic genes, accompanied by increased levels of both precursor and mature sterol regulatory element-binding protein 1 (SREBP-1) with prominent nuclear localization. In three-dimensional spheroid cultures, FKD cells exhibited extensive vacuolar degeneration, reduced LC3B expression, and p62 accumulation, whereas LAMP-1 expression remained unchanged. Low-vacuum scanning electron microscopy further revealed rough, granular deposits on the spheroid surface, suggesting structural deterioration associated with impaired autophagy. Treatment with the autophagy inducer Tat-Beclin1 partially restored LC3B expression and attenuated lipid droplet accumulation. Collectively, these findings indicate that Fbxw7 plays a critical role in maintaining hepatocellular homeostasis through coordinated regulation of lipid metabolism and autophagic degradation. Loss of Fbxw7 disrupts this balance, leading to characteristic metabolic and morphological alterations in hepatocellular carcinoma cells.
Autoimmune hepatitis (AIH) lacks a single disease-specific diagnostic test, particularly in acute-onset or seronegative cases with normal IgG levels. Recent progress in high-throughput autoantibody profiling has enabled systematic and unbiased identification of novel autoantibodies, offering new insights into disease diagnosis and underlying pathogenic mechanisms. This review summarizes emerging autoantibodies in AIH, primarily focusing on antibodies identified through proteome-wide approaches. We highlight anti-docking protein 2 (DOK2) antibodies, identified using a human protein microarray and validated by ELISA, immunohistochemistry, and imaging mass cytometry. These antibodies exhibit high diagnostic performance, including in patients with IgG-normal and ANA-negative AIH, and are correlated with disease activity. Spatial analyses revealed that DOK2 was expressed predominantly in hepatic macrophages and subsets of T cells, providing mechanistic insight into immune regulation in AIH. These findings illustrate how integrative serological and molecular morphology approaches can refine AIH diagnosis and deepen the understanding of disease pathogenesis.
Biliary tract carcinoma (BTC) is a malignancy with poor prognosis. Current molecular targeted therapies benefit only a limited subset of patients, underscoring the need for novel approaches, such as combining DNA-damaging chemoradiotherapy with an immune checkpoint inhibitor. Although cellular responses following chemoradiotherapy-induced DNA damage are essential, they remain poorly understood in the context of three-dimensional tumor structures. This study aimed to investigate the effects of ionizing radiation, a DNA-damaging cancer therapy, on patient-derived BTC organoids. DNA repair and gene expression regulation was integrally examined by immunofluorescence and RNA-Seq analysis following 10 Gy of X-rays. X-ray irradiation caused significant morphological changes. In addition, X-ray irradiation upregulated extracellular matrix-related gene expression as revealed by RNA sequencing. DNA damage response analysis indicated that non-homologous end joining was the primary repair pathway in patient-derived BTC organoids. Moreover, X-ray irradiation activated immune-related pathways, such as cGAS/STING, RIG-I/MDA-5, and JAK/STAT, suggesting potential immune activation following radiotherapy. Our study revealed that DNA damage response altered the tumor structure and modulated the expression of multiple genes, including extracellular matrix- and immune-related genes. Studies using in vitro organoid models can be useful for investigating three-dimensional cellular responses after DNA-damaging cancer therapies, such as chemoradiotherapy.
Neuroendocrine carcinoma and high-grade serous ovarian cancer (HGSCO) form a rare mixed ovarian tumor. Ovarian cancer is the most lethal gynecological malignancy, with HGSCO being the most common (60–70
This study evaluated the utility of rapid liquid-based cytology with a shortened fixation time for intraoperative effusion cytology compared to conventional smears. Despite widespread use, intraoperative applications of liquid-based cytology are understudied; no quantitative paired comparison of rapid liquid-based cytology and conventional smears under identical constraints has been reported. We therefore tested whether rapid liquid-based cytology enriches diagnostically informative cells during time-limited reads, supporting more reliable reporting. We retrospectively analyzed 67 effusion (17 adenocarcinoma-positive and 50 adenocarcinoma-negative) samples collected between May, 2019 and February, 2024. Smear area, nucleated cell counts per field and slide, atypical cell counts, and atypical cell aggregates were quantified for paired preparations and compared using the Wilcoxon signed-rank test. Rapid liquid-based cytology yielded a markedly small smear area with significantly high nucleated cell counts per field, and conventional smears yielded high total nucleated cell counts per slide. Rapid liquid-based cytology also revealed significantly high numbers of atypical cells and aggregates. Subgroup analyses based on specimen characteristics (mesothelial-dominant, blood-dominant, and mixed) confirmed that rapid liquid-based cytology yielded high per-field nucleated cell counts in all groups. These results suggest that rapid liquid-based cytology concentrates diagnostic cells into a small standardized smear area, supporting timely reporting.
We retrospectively analyzed 116 cases of colorectal cancer (CRC) to evaluate the relationship between nutritional indicators (NIs) and clinicopathological features, including immune cell infiltration in the tumor microenvironment. The geriatric nutritional risk index (GNRI), prognostic nutritional index (PNI), modified Glasgow prognostic score (mGPS), Controlling Nutritional Status (CONUT) score, and neutrophil-to-lymphocyte ratio (NLR) were assessed. Immune cell densities (CD3, CD8, Foxp3, Iba1, CD163) were quantified by immunohistochemistry. Poor NIs were associated with older age, lower body mass index, advanced tumor stage, elevated tumor markers, and reduced immune cell infiltration. Specifically, low GNRI and high mGPS correlated with reduced Foxp3-positive cells; low PNI with reduced Foxp3- and CD20-positive cells; and high NLR with reduced CD3-, CD8-, CD20-, and Iba1-positive cells. The GNRI- and mGPS-abnormal groups showed significantly reduced Foxp3-positive cells, and the CONUT-abnormal group exhibited decreased CD3- and CD20-positive cells. Patients with abnormal NLR had significantly worse cancer-specific survival. However, it was not identified as an independent prognostic factor in the multivariate analysis. Nutritional impairment in CRC patients is linked not only to poor clinical outcomes, but also to a tumor-promoting microenvironment characterized by diminished immune cell infiltration. Assessing NIs may help guide nutritional and therapeutic strategies to improve prognosis.
Although systemic lupus erythematosus (SLE) can affect multiple organ systems, manifestations within the digestive tract are not typically conspicuous during the early phase of the disease. The liver damage caused by SLE is mild and insidious. Patients with SLE presenting with acute liver failure as the primary manifestation are significantly rarer in clinical diagnosis. The absence of diagnostic criteria for digestive system manifestation in SLE complicates the diagnosis of lupus as the underlying cause, particularly during the initial presentation. Timely recognition of the disease and commencement of immunosuppressive treatment are crucial for enhancing clinical outcomes. This article reports a rare case of SLE in a 9-year-old Chinese girl presenting with acute liver failure as the initial symptom. The child lacked typical lupus features such as skin and joint manifestations, making the initial diagnosis extremely challenging. The diagnosis relied on key liver pathological examinations and positive serological lupus-specific antibodies, and gene sequencing identified the c.740C > T (p.A247V) mutation of the TREX1 gene. After treatment, her condition improved. This case highlights the importance of early immunological and genetic screening in pediatric patients with unexplained acute liver failure to identify potential SLE. This case represents the first reported instance of pediatric SLE presenting with acute liver failure as the initial manifestation, associated with the c.740C > T (p.A247V) mutation. These findings highlight the critical importance of a multi-modal diagnostic approach—encompassing immunological, pathological (biopsy), and genetic assessments—for the evaluation of children with such atypical presentations.
Dry eye is a multifactorial ocular surface disease that may be accompanied by visual impairment and ocular surface damage. Thrombospondin 1 (THBS1) was found to be highly expressed in corneal epithelial cells of dry eye mouse models. Our research aimed at exploring the role and regulatory mechanism of THBS1 in dry eye mouse models. Both eyes of mice with benzalkonium chloride (BAC)-induced dry eye were subconjunctivally injected with the recombinant adeno-associated virus (AAV) vector containing the THBS1 silencing plasmid. Under a slit lamp biomicroscope, the conjunctival irritation including edema, hyperemia, and secretion was scored. Fluorescein staining was performed to evaluate corneal epithelial damage. The conjunctiva tissues were obtained for ELISA, RT-qPCR, histological staining, and western blotting. Dry eye model mice exhibited severe ocular surface damage, reduced tear secretion, conjunctival goblet cell loss, increased conjunctival inflammation, elevated THBS1 expression, and the TGF-β/NLRP3 inflammasome pathway activation. THBS1 silencing ameliorated ocular surface damage, increased tear secretion, attenuated conjunctival goblet cell loss, mitigated conjunctival inflammation, and repressed the TGF-β/NLRP3 inflammasome pathway activation in dry eye mice. THBS1 silencing protects mice against dry eye by inhibiting TGF-β/NLRP3 inflammasome-mediated inflammatory response.
We report a rare case of thymoma with extensive clear cell components, and discuss the histogenesis of clear cell transformation. Although thymic clear cell carcinoma is classified as a subtype of thymic carcinoma, thymoma with extensive clear cell components has not been recognized as a subtype in the World Health Organization classification (2021). Therefore, such thymomas may be erroneously misdiagnosed as clear cell carcinoma. Thymic clear cell carcinomas are positive for glycogen, with a high Ki67 index, whereas the clear cell component of the present tumor was negative, with a low index; this difference may help to distinguish thymomas with extensive clear cell components from thymic clear cell carcinomas. In this tumor, spindle cells predominantly proliferated in the center, whereas clear cells were densely distributed in the periphery. Hence, the initial tumor likely consisted of spindle cells, some of which transformed into clear cells as the tumor grew larger. In addition, nuclei of the clear cells were occasionally positive for autophagy-related 4 cysteine peptidase. Electron microscopically, clear cells with eccentrically located nuclei and cytoplasm contained large vacuoles, in which irregularly shaped membranous structures were observed. The histology, immunohistochemistry, and ultrastructure suggest that clear cell transformation represents a type of cellular degeneration.
