Mutations in TP53 occur in approximately half of all common cancer types and play a critical role in tumor progression and metastasis. However, because mutant TP53 can induce both loss- and gain-of-function effects, the development of drugs that directly target TP53 remains challenging. To identify downstream effectors of specific TP53 mutations, we compared gene expression profiles of colorectal (CRC), lung (LC), and stomach cancers (STC) carrying wild-type or mutant TP53. TP53 mutations were frequent (CRC 70.9%, LC 50.1%, STC 45.7%), with TP53-R273H being the most common. Comprehensive analyses comparing patients with TP53 mutations and those with wild-type TP53 identified 24 genes whose expression was significantly increased in TP53-mutated patients, and 20 genes whose expression was decreased across all three tumor types. Among them, PABPC1 showed significantly increased expression in A549 lung cancer cells and MCF7 breast cancer cells with wild-type TP53 transfected with TP53-R273H mutants. Silencing PABPC1 suppressed proliferation of TP53-R273H-mutant colon cancer cells (HT29, SW480). Additionally, patients with CRC harboring the TP53-R273H with high PABPC1 expression had poorer overall survival than those with low PABPC1 expression. These findings indicate that TP53-R273H upregulates PABPC1, suggesting that PABPC1 may function as an oncogene and represent a promising therapeutic target in patients with TP53-R273H mutation.

The tubarial gland is a potential salivary organ in humans. In this study, the innervation of the tubarial gland was examined using immunohistochemistry for protein gene product 9.5 (PGP 9.5), dopamine β-hydroxylase (DBH), choline acetyltransferase (ChAT), and vasoactive intestinal polypeptide (VIP). PGP 9.5-positive nerve fibers displayed thick and slender shapes around the mucous and serous acinar cells, respectively, in the tubarial gland. Both mucous and serous acinar cells were enclosed by thick and fine PGP 9.5-positive nerve fibers in major salivary glands. Other neurochemical substances were usually present in fine and varicose nerve fibers in glandular acini. DBH-positive nerve fibers were infrequent, and VIP-positive nerve fibers were rich in the tubarial gland, compared with submandibular and parotid glands. In the tubarial gland, the acinar cells were enclosed by a few DBH-positive nerve fibers and many ChAT- and VIP-positive nerve fibers. Serous acini had sparse DBH-positive nerve fibers and dense VIP-positive nerve fibers in the tubarial gland compared with submandibular and parotid glands. The parasympathetic ganglion probably sends the ChAT- and VIP-positive nerve fibers to the tubarial gland. Acetylcholine and VIP in the gland may contribute the immune system by regulating moisture, ciliary movement, and immune proteins in the respiratory tract.

The elimination of self-reactive clones through B cell receptor (BCR)-induced apoptosis is a fundamental mechanism for establishing self-tolerance in immature B cells. We previously reported that protein phosphatase 6 (PP6) suppresses BCR-induced apoptosis in the immature B cell line WEHI-231 by maintaining the expression of the anti-apoptotic protein, Bcl-xL. Immunoglobulin-binding protein 1 (IGBP1) interacts with the catalytic subunit of PP6 (PP6c) to regulate its phosphatase activity. In this study, we investigated the role of IGBP1 in BCR-mediated apoptosis. We found that IGBP1 overexpression significantly suppressed anti-IgM-induced apoptosis in WEHI-231 cells, whereas its siRNA-mediated knockdown accelerated cell death. Co-immunoprecipitation and activity assays revealed that BCR stimulation attenuates the IGBP1-PP6c interaction and reduces PP6 phosphatase activity. Importantly, this reduction in PP6 activity was prevented by IGBP1 overexpression and exacerbated by its knockdown. Furthermore, IGBP1 positively regulated Bcl-xL stability following BCR stimulation. Notably, exogenous overexpression of Bcl-xL effectively rescued the enhanced apoptosis observed in IGBP1-knockdown cells. Collectively, our findings demonstrate that IGBP1 protects immature B cells from BCR-induced apoptosis by sustaining the IGBP1-PP6c complex and its phosphatase activity, thereby maintaining the expression of Bcl-xL.

Gastric cancer exhibits substantial intratumoral heterogeneity; however, genetic differences between spatially separated differentiated and undifferentiated components within the same tumor, particularly in deficient mismatch repair (dMMR) cases, remain poorly defined. We compared somatic mutation and copy number variation (CNV) profiles between these two histologic types within the same gastric tumor, including dMMR cases. Formalin-fixed, paraffin-embedded specimens from five gastric cancers were macrodissected to separate differentiated and undifferentiated components and analyzed using a 225-gene targeted panel. All cases shared one or more somatic mutations between the two components, consistent with a common clonal origin. However, most mutations were type-specific, with greater divergence in dMMR tumors. dMMR cases harbored more mutations than proficient mismatch repair cases, and many were detected at low variant allele frequency; type-specific low-variant allele frequency mutations were enriched in the undifferentiated component. CNV patterns also differed between the two components, and most CNVs were amplifications. NOTCH1 CNV was shared across both components in all cases, whereas CNVs overall were more frequent in the differentiated component. These findings indicate that differentiated and undifferentiated components within the same gastric tumor share truncal mutations but accumulate distinct additional alterations, underscoring spatial heterogeneity and supporting multiregion sampling for molecular profiling.

This study aimed to elucidate the mechanism underlying tolerance to the sedative effects of propofol, focusing on cannabinoid 1 (CB1) receptor regulation. Male Wistar rats received continuous propofol infusion (30-40 mg/kg/h) for 1, 2, 4, or 12 h prior to tissue collection. Brain regions including the cortex, hippocampus, striatum, brain stem, and cerebellum were analyzed. Changes in CB1 receptor mRNA transcript abundance were assessed using semi-quantitative PCR. Across all examined regions, CB1 receptor mRNA levels showed a time-dependent decrease following propofol administration. Reductions were detectable as early as 1 h and became more pronounced at 4 and 12 h. This downregulation was consistently observed across multiple brain regions, suggesting a widespread effect rather than region-specific modulation. These findings suggest that continuous propofol exposure leads to significant downregulation of CB1 receptor expression, which may contribute to receptor desensitization and the development of tolerance to its sedative effects. Modulation of the endocannabinoid system may therefore play a key role in propofol-induced tolerance.

Plasminogen activator inhibitor-1 (PAI-1) is a serine protease inhibitor that controls trophoblast invasion by inhibiting fibrinolysis at the feto-maternal interface. This study aimed to compare PAI-1 expression patterns in decidual cells between different abortion types and investigate the role of PAI-1 in early pregnancy. Missed abortion (n = 22) and incomplete abortion (n = 16) cases were compared with induced abortion serving as normal pregnancy control (n = 16). PAI-1 localization and expression levels were evaluated using immunohistochemistry. PAI-1 was distributed in the cytoplasm of decidual cells. In addition, many PAI-1-positive cells were observed in the decidua basalis just under the Nitabuch fibrinoid layer, where many extravillous trophoblasts (EVTs) invaded. The percentage of PAI-1-positive decidual cells was not reduced in missed abortion (49.7 ± 29.6%) but was significantly reduced in incomplete abortion (11.6 ± 12.6%) compared to induced abortion as control (44.4 ± 10.9%). The significant reduction of PAI-1-positive decidual cells in incomplete abortion, which is characterized by bleeding, suggests that PAI-1 deficiency may cause excessive fibrinolysis leading to this clinical outcome.

Many scientists are interested in anti-aging interventions and seek to identify anti-aging medicines. In our previous study, we reported that an antitussive drug, tipepidine (TIP), activates AMP-activated protein kinase (AMPK) and improves glucose intolerance in high-fat diet-induced obese mice. Activation of AMPK has been reported to extend lifespan in various experimental organisms, including Caenorhabditis elegans. In this study, we examined whether TIP extends lifespan in C. elegans via AMPK activation. TIP treatment (50-100 μM) extended the median lifespan of wild-type C. elegans (N2), but not that of the AMPK catalytic subunit mutant aak-2(rr48). These results suggest that TIP extends lifespan in C. elegans in an AMPK-dependent manner.

The cGAS-STING pathway is an innate immune pathway that senses microbial and host-derived double-stranded DNA in the cytosol, and triggers type I interferon responses and proinflammatory cytokine production. Recently, activation of the cGAS-STING pathway in tumors is reported to enhance anti-tumor effects, while it is often inactivated in tumors. Therefore, to develop strategies to activate STING in tumors has become imperative in cancer therapy. Here we report that the cGAS-STING signaling is inactivated in DU145 prostate cancer cell line and find that the inactivation mechanism is distinct from the previously reported mechanisms such as DNA methylation. Treatment of DU145 cells with a lysosomal inhibitor Baf-A1 increased a STING protein level. Knockdown of VPS4, the essential factor in lysosomal microautophagy, suppresses STING degradation. Most importantly, knockdown of cGAS, the enzyme responsible for the production of the endogenous STING ligand cGAMP, fully restores a STING protein level and a response to a synthetic membrane-permeable STING agonist MSA-2. Thus, these results suggest that the constant cGAS activation causes the lysosomal degradation of STING and underlies the unresponsiveness of the STING signaling in DU145 cells. Our results may lead to new strategies to enhance STING-targeted cancer immunotherapy.

B cell antigen receptor (BCR) signaling plays a critical role in regulating B cell fate, including activation, tolerance, and apoptosis. In immature B cells, strong BCR engagement induces apoptosis, which contributes to the elimination of autoreactive clones during negative selection. However, the molecular mechanisms linking BCR signaling to apoptotic pathways remain incompletely understood. In this study, we investigated the role of Ca2+/calmodulin-dependent protein kinase II (CaMKII) in BCR-induced apoptosis using the immature B cell line WEHI-231. Stimulation of the BCR with anti-IgM antibody induced a rapid increase in intracellular Ca2+ levels and promoted apoptosis in WEHI-231 cells. BCR engagement also induced phosphorylation of CaMKII, indicating activation of this kinase downstream of Ca2+ signaling. Pharmacological inhibition of CaMKII with KN-93, a CaMKII inhibitor, attenuated BCR-induced apoptosis, whereas overexpression of CaMKII enhanced cell death. We further found that BCR stimulation resulted in downregulation of the anti-apoptotic protein Bcl-xL, and inhibition of CaMKII prevented this reduction. Conversely, CaMKII overexpression further enhanced Bcl-xL downregulation following BCR stimulation. Importantly, restoration of Bcl-xL expression significantly rescued CaMKII-mediated apoptosis. These findings identify a Ca2+-CaMKII-Bcl-xL signaling axis that promotes BCR-induced apoptosis in immature B cells.

Chromosomes undergo dynamic structural reorganization during the cell cycle and become highly condensed during mitosis. This condensation process involves the assembly of chromatin fibers, in which chromosome-associated proteins (e.g., condensin) and post-translational modifications (e.g., histone deacetylation) play key roles. Although an increasing number of studies have advanced our understanding of chromatin fiber assembly into chromosomes, detailed morphological information on chromatids remains limited. In this report, we used atomic force microscopy to examine the nanoscale structure of chromosomes preserved in liquid and found that mitotic chromosomes are composed of a series of thin filamentous structures. Quantitative analysis of chromosomes from the chromokinesin KIF4A-depleted cells and under histone deacetylase inhibition revealed that the filament width and inter-filament spacing are regulated by KIF4A and histone deacetylation, likely reflecting their specific contributions at different hierarchical levels of chromosome assembly. These findings demonstrate that atomic force microscopy enables the detection of fine nanoscale structural alterations beyond the resolution of optical microscopy and provides new morphological insights into the organization of mitotic chromosomes.

Disuse induces capillary regression in skeletal muscle, which is associated with excessive production of reactive oxygen species (ROS), impaired mitochondrial oxidative capacity, and a shift from slow- to fast-twitch muscle fibers. Green tea is rich in catechins with potent antioxidant properties. We hypothesized that compressed residual green tea extract, a byproduct of green tea manufacturing, may attenuate capillary regression during muscle disuse. Twenty female Wistar rats were randomly assigned to four groups: control (CON), compressed residual green tea supplementation (GTs), hindlimb unloading (HU), and hindlimb unloading with compressed residual green tea supplementation (HU+GTs). Hindlimb unloading was performed for 2 weeks. Compressed residual green tea extract (850 mg/kg/day) or distilled water was administered orally. HU induced significant reductions in soleus muscle mass, capillary-to-fiber (C/F) ratio, slow-twitch fiber proportion, and succinate dehydrogenase (SDH) activity, along with increased ROS production. Compressed residual green tea supplementation significantly attenuated capillary regression, preserved slow fiber composition, restored SDH activity, and suppressed ROS overproduction under unloaded conditions. These findings suggest that compressed residual green tea may serve as a practical nutritional strategy to maintain skeletal muscle capillarity during disuse by preserving oxidative fiber phenotype and mitochondrial function.

Intracerebral hemorrhage (ICH) is a devastating form of stroke with limited treatment options. Although exercise is beneficial in ischemic stroke, its preconditioning effects in hemorrhagic stroke remain unclear. We examined whether voluntary wheel running prior to ICH modulates acute outcomes and gene expression. Male ICR mice underwent three weeks of voluntary running or sedentary housing before ICH induction. Neurological function was assessed 3 days post-ICH, and perihematomal tissue was analyzed by quantitative PCR. ICH increased expression of Iba1, GFAP, C3, MMP9, TIMP1, and Caspase3, while reducing BDNF expression. Exercise preconditioning did not improve acute behavioral deficits at the group level. However, running distance strongly predicted molecular and pathological responses. BDNF expression correlated positively with exercise volume, whereas C3 showed a negative correlation, with a similar trend for GFAP. Caspase3 and Bax expression were negatively correlated with running distance. Although hematoma volume did not differ significantly between groups, greater running distance was associated with smaller hematoma volume within the exercise group. These findings indicate that habitual voluntary exercise does not attenuate acute functional deficits after ICH but induces dose-dependent molecular adaptations related to neuroplasticity, glial inflammation, and cell death, potentially priming the brain for enhanced recovery in later phases.

Ultrastructural analysis has declined in diagnostic pathology since the advent of immunohistochemistry for formalin-fixed paraffin-embedded tissues. To reevaluate the utility of ultrastructural analysis, we adopted the osmium maceration method for scanning electron microscopy (SEM) of formalin-fixed surgical specimens: renal oncocytoma, chromophobe renal cell carcinoma (ChRCC), and tumors categorized as other oncocytic tumors of the kidney (OOT). Light microscopy and immunohistochemistry have revealed that these tumors have overlapping features. SEM observation of the formalin-fixed tumors highlighted distinct morphological features: The oncocytomas contained large mitochondria with vesicular cristae; ChRCCs harbored small mitochondria with microvesicles, and OOTs showed abundant but small mitochondria with lamellar cristae. Quantitative morphometry confirmed that the number of mitochondria in oncocytomas and OOTs exceeded that of non-neoplastic lesions, whereas ChRCCs contained fewer mitochondria. In addition, mitochondria in ChRCCs and OOTs were smaller than those of non-neoplastic lesions, whereas mitochondria in oncocytomas were larger. Thus, SEM of formalin-fixed tissues, optimized by osmium maceration, can provide diagnostically valuable information beyond conventional histology and immunohistochemistry that may help to clarify the biology of challenging oncocytic and chromophobe renal tumors.

Despite advances in oral cancer treatment, therapeutic resistance remains a major challenge. Cancer-associated fibroblasts (CAFs) play a pivotal role in shaping the tumor microenvironment. Building on our previous findings on immune responses in gingival fibroblasts, we investigated the immune characteristics of CAFs. Myofibroblast CAF-like cells (myCAF-like) were established using a co-culture system and compared with gingival fibroblast-derived myofibroblasts. MyCAF-like cells exhibited markedly reduced expression of TLR3 and TLR4, whereas RIG-I, COX-2, IL-1β, IL-6, and IL-8 were significantly upregulated. Activation of p38 MAPK signaling contributed to the increased expression of COX-2 and pro-inflammatory cytokines, and TGF-β was involved in the phenotypic transformation of gingival fibroblasts into myCAF-like cells. Microarray analysis revealed upregulation of IL-1α, IL-1β, CXCL8, PTGS2, and CXCL5, alongside downregulation of OMD, COL15A1, and ASPN in myCAF-like cells. Collectively, these findings demonstrate that CAFs acquire a distinct inflammatory signature that differs from gingival fibroblasts and promotes tumor invasion and progression. Targeting CAF-associated inflammatory and signaling pathways may represent a promising strategy to overcome tumor infiltration and treatment resistance in oral cancer.

This study aimed to elucidate the relationship between the spatiotemporal behavior of macrophages during exercise and axonal regeneration following peripheral nerve injury. A sciatic nerve crush model was created using male C57BL/6J mice, which were divided into sedentary and exercise groups. The exercise group performed low-intensity treadmill running (10 m/min, 60 min/day, 5 days/week) starting 3 days post-injury. Immunohistochemical analyses of Neurofilament 200 kDa (NF200), Growth Associated Protein-43 (GAP-43), F4/80, and arginase-1 (Arg-1), a representative marker of M2 macrophages and a downstream effector of the JAK-STAT6 signaling pathway, were conducted in the proximal and distal regions of the injured nerves. Functional recovery was assessed using the sciatic functional index (SFI) and compound muscle action potential (CMAP). At 7 and 14 days post-injury, the exercised group exhibited a significant increase in axonal number and Arg-1-positive area, specifically in the distal region. SFI and CMAP analyses also demonstrated enhanced functional recovery in the exercise group. These findings suggest that low-intensity treadmill exercise may enhance axonal regeneration, potentially through transient activation and local accumulation of M2-polarized macrophages. Therefore, exercise-induced regulation of macrophage dynamics may represent a novel therapeutic strategy for peripheral nerve repair.