Cytojournal最新文献

Objective: Renal cell carcinoma (RCC) is a prevalent malignant tumor of the kidney that has considerable heterogeneity and intricate molecular pathways. Incidence and death rates associated with kidney cancer have been increasing steadily. Consequently, investigating its molecular basis and identifying new biomarkers for targeted therapies have become important. Ubiquitin C-terminal hydrolase L5 (UCHL5) has an effect on tumor initiation and progression. This study aims to explore the role of UCHL5 in promoting RCC development and identify related molecular mechanisms.

Material and methods: Quantitative real-time polymerase chain reaction and Western blot (WB) analyses were employed to measure UCHL5 expression. Stable UCHL5 overexpression and knockdown cell lines were generated using lentiviral transfection in 786-O cells. Colony formation and 5-ethynyl-2'-deoxyuridine assays were used to assess cell proliferation. Cell migration was evaluated using scratch assays, while invasion capacity was determined by Transwell assays. Commercial kits were utilized to quantify glucose consumption and lactate production. WB was applied to detect proteins related to glycolysis and components of the Phosphoinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway. The function of UCHL5 in RCC cells was validated in vivo through an animal tumor model.

Results: The expression of UCHL5 increased in RCC cell lines (P < 0.01). Silencing UCHL5 significantly impaired the malignant behavior of RCC cells (P < 0.01), whereas its overexpression promoted this cellular behavior (P < 0.05). Reducing UCHL5 levels decreased glucose uptake, lactate secretion, and protein expression involved in glycolysis, while overexpression enhanced glycolytic activity in RCC cells (P < 0.01). Knockdown of UCHL5 diminished the phosphorylation of PI3K/AKT/mTOR pathway proteins (P < 0.05), whereas the opposite effect was observed upon overexpression (P < 0.01). Treatment with 740Y-P effectively reversed the suppression of glycolysis, proliferation, and metastasis caused by UCHL5 knockdown (P < 0.01). Conversely, LY294002 inhibited the glycolytic enhancement and aggressive phenotypes induced by UCHL5 overexpression (P < 0.01). Animal experiments further confirmed that UCHL5 downregulation suppressed RCC growth through the PI3K/AKT/ mTOR pathway (P < 0.01).

Conclusion: UCHL5 activates the PI3K/AKT/mTOR cascade, which enhances the glycolysis of RCC cells and promotes the development of renal cancer. This study provides insights into the molecular mechanisms underlying the oncogenic role of UCHL5 in RCC.

Objective: Despite the availability of many epilepsy drugs, certain epilepsy patients still cannot be treated with medication. The dysfunction of oxoglutarate dehydrogenase-like (OGDHL) is related to neurodegeneration. This article aimed to explore the role of OGDHL on interleukin (IL)-1β-induced epileptic cell model and its molecular mechanism.

Material and methods: An epileptic cell model was established using IL-1β. Quantitative real-time polymerase chain reaction was used to detect the expression levels of tumor necrosis factor-α, IL-1β, and IL-6 after different treatments. Western blot was used to detect the recovery effect of OGDHL overexpression on the IL-1-induced epilepsy model of CTX-TNA cells through the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway and collagen type IV alpha 2 (COL4A2) expression level. Meanwhile, the hypertrophy, viability, and apoptosis of CTX-TNA cells were = evaluated through terminal deoxynucleotidyl transferase dUTP nick end labeling staining, cell counting kit-8 assay, and glial fibrillary acidic protein expression. In addition, we evaluated the effect of the JAK/STAT signaling pathway agonist α7nAchR on the effects of OGDHL overexpression. The influence and possible mechanisms of OGDHL overexpression were comprehensively assessed through the above experimental methods.

Results: Our results suggest that OGDHL overexpression can alleviate IL-1β-induced inflammatory response to epilepsy through the JAK/STAT signaling pathway and significant upregulation of COL4A2 expression level (P < 0.001). In addition, ODGHL overexpression can regulate the hypertrophy and apoptosis of CTX-TNA cells (P < 0.001). The effect of OGDHL overexpression on reducing epileptic inflammatory response was further demonstrated through the intervention of α7nAchR, which serves as an agonist in the JAK/STAT signaling pathway.

Conclusion: ODGHL overexpression may inhibit the JAK/STAT signaling pathway by upregulating COL4A2 expression, and it inhibited the IL-1β-induced inflammation of CTX-TNA cells in an epileptic model.

Radiation changes in lung cytology specimens can lead to diagnostic challenges, especially during on-site evaluation. We present a case of two patients, both with a prior history of lung malignancy and subsequent radiation treatment, one with a metastatic high-grade undifferentiated pleomorphic sarcoma and the other with a small cell carcinoma. One was thought to be malignant, and one was thought to be atypical on-site. Common to both cases was a hypocellularity and a prominent repair-like/reactive change with polygonal cells or elongated fiber-like cells and dense cytoplasm mimicking atypical squamous cells in shape and cytoplasm. Knowledge of these prominent findings may help cytopathologists quickly recognize radiation changes at on-site evaluation and avoid overcalls.

Objective: Tumor-infiltrating lymphocyte (TIL) assessment is recognized as an important prognostic and therapeutic biomarker in several tumors, including triple-negative breast cancer (TNBC). TILs face several challenges, including low to intermediate interobserver concordance. The use of tissue microarray (TMA) in TIL assessment and its relationship/concordance with whole sections (WS) has been studied only rarely. The aim of this study is to evaluate the reproducibility of TIL assessment in TNBC among various users and to compare TIL assessment using TMA versus whole tissue sections (WS).

Material and methods: Hematoxylin and eosin-stained sections of TMAs were prepared and assessed for quality and adequacy. Corresponding WS slides for all cases were retrieved and independently scored for TILs by one junior and two senior pathologists. The assessment followed the guidelines of the International TILs Working Group. TIL scores were categorized into three groups: Low: <20%; intermediate: 20-49%; and high: ≥50%. A total of 100 cases were collected; however, 76 cases were evaluable after excluding inadequate samples or cases with missing information.

Results: Fleiss' kappa was used to assess interobserver agreement among the three raters for both WS and TMA datasets. The Intraclass Correlation Coefficient (ICC) was used to assess the degree of agreement among the three raters for continuous variables before categorization, and Cohen's kappa was applied to both WS and TMA datasets to assess interobserver agreement between any two raters. The interobserver agreement for WS analysis (Cohen's kappa) was fair, while for TMA, it was moderate. The ICC also showed moderate agreement. Cohen's kappa for TMA versus WS for each of the three observers ranged from fair to strong. Fleiss' kappa showed fair agreement for WS versus TMA.

Conclusion: Interobserver variability in TIL scoring remains a challenge. TMAs improve consistency but at the cost of reduced correlation with WS assessments.

Objective: In cytological evaluation, atypical cells are often recognized by comparing them to presumed "normal" counterparts and judging the extent of morphological deviation. However, there is no clear or universally accepted definition of what constitutes a "normal cell," and the assessment often relies on the examiner's subjective judgment. This paper aims to establish the reference values for the cytological evaluation of the tongue mucosa by quantitatively analyzing nuclear-to-cytoplasmic ratios (NCRs) and the clarity of the features of both nuclear and cytoplasm.

Material and methods: Cytological specimens from seven patients (three males and four females; average age, 62.4 years old) with tongue mucosal lesions were collected (with the Papanicolaou [Pap] stain). Acquired images were analyzed using ImageJ software. We quantified NCRs and brightness values (BV). Statistical analysis was performed using the Steel-Dwass test.

Results: Significant differences in NCR and nuclear BV were observed across Pap Classes 1, 3, and 5. Reference values for normal cells were defined as a mean NCR of 0.031 ± 0.015 for Class 1, 0.074 ± 0.048 for Class 3, and 0.307 ± 0.170 for Class 5. The nuclear BV range widened with increasing Class: 68.90-145.16 (Class 1), 49.91-163.40 (Class 3), and 28.11-183.54 (Class 5).

Conclusion: Our findings suggest that NCR and nuclear brightness can be used as objective criteria for evaluating tongue mucosal cells. For the detection and diagnosis of oral malignant disorders in early stage, the result is a kind of criterion.

Gastrointestinal stromal tumors (GISTs) represent the most common mesenchymal neoplasms of the gastrointestinal tract and are typically associated with activating mutations in the kinase insert domain receptor (c-KIT) or platelet-derived growth factor receptor alpha. The advent of targeted therapies, such as imatinib, has substantially improved clinical outcomes; however, primary double-mutant GISTs present significant therapeutic challenges. We report the case of a 51-year-old man who presented with a 2-week history of left upper abdominal pain and melena. Contrast-enhanced abdominal computed tomography revealed a mass in the pelvic region of the small intestine. Histopathological analysis demonstrated a spindle cell morphology with a high mitotic index. Immunohistochemical staining was positive for CD117, CD34, and Dog-1, confirming the diagnosis of a high-risk small intestinal GIST. At the time of diagnosis, genetic testing was not performed, and the patient was initiated on imatinib therapy. After 5 years of treatment, the patient developed clinical resistance. First-generation sequencing identified concurrent mutations in c-KIT exons 11 (V560D) and exon 17 (N822K), implicating these double mutations in acquired imatinib resistance. This case underscores the clinical significance of double mutations in GIST, the limitations of first-line therapy in such contexts, and the importance of early genetic profiling to inform personalized treatment strategies.

Objective: As the first gynecological tumor, ovarian cancer seriously threatens women's lives and health. Here, the possible mechanism of midazolam against ovarian cancer progression was investigated.

Material and methods: OVCAR3 and SKOV3 cells were treated with midazolam for 24 h, and cell activity was subsequently detected by cell counting kit-8 assay to screen for the suitable treatment concentrations of midazolam. Cell proliferation was investigated through 5-ethynyl-2'-deoxyuridine staining and cell plate cloning experiments. Apoptosis rate was detected by flow cytometry, and cell invasion and migration ability were examined through transwell test and scratch test. Western blot was adopted to explore the expression changes of apoptotic proteins, cell cycle-related proteins, and extracellular signal-regulated kinase/c-Jun N-terminal kinase (ERK/JNK) pathway-related proteins.

Results: After midazolam intervention, the OVCAR3 and SKOV3 cells showed decreased proliferation, invasion, and migration abilities and accelerated apoptosis rate. Western blot results displayed that midazolam downregulated the phosphorylation levels of ERK, JNK, and P38. Anisomycin reversed the effect of midazolam on the ERK/JNK pathway and cell proliferation, apoptosis, cell cycle, invasion, and migration.

Conclusion: Midazolam can block the development of ovarian cancer, and the relevant mechanism may be realized through the ERK/JNK pathway.

Objective: Hypoxia-inducible factor 1 (HIF-1) signaling mediates multiple links of wound healing. Tissue hypoxia and dysregulation of HIF-1 signal play a crucial role in non-healing diabetic wounds. Previous studies have found that roxadustat (FG-4592) can promote epidermal stem cell proliferation by upregulating the HIF signaling pathway. This study aimed to investigate the role of roxadustat in the wound healing of diabetic mice.

Material and methods: The study was divided into in vivo and in vitro experiments. In the in vivo experiment, mice were categorized into three groups: control group, diabetes group, and diabetes + roxadustat group. Diabetic mice were injected intraperitoneally with roxadustat daily at a dose of 10 mg/kg. Hematoxylin & eosin staining and Masson staining were employed to assess wound healing speed and quality. Immunohistochemical staining was used to detect HIF-1a and proliferating cell nuclear antigens. Western blot was conducted to examine markers associated with Notch1 signaling pathway activation (Notch Intracellular Domain [NICD]), keratinocyte differentiation, and angiogenesis. In the in vitro experiment, HaCaT cells were divided into control (Glu 5.5 mM), high-glucose (Glu 30 mM), and high-glucose + drug (Glu 30 mM + FG-4592) groups, with a treatment concentration of FG-4592 set at 10 µM. Following 48 h of treatment period, protein was extracted for co-immunoprecipitation analysis to determine the interaction between HIF-1a and NICD, and fluorescence staining was conducted to assess their co-localization.

Results: Roxadustat reversed the slow wound healing caused by diabetes and significantly improved the quality of healing (P < 0.05). It upregulated the inhibited HIF-1 signaling in diabetic mice (P < 0.05) and triggered cell proliferation. It downregulated the hyperactivated Notch1 signaling in diabetic mice (P < 0.05) and induced keratinocyte dedifferentiation, which were both responsible for wound re-epithelialization. Roxadustat also reversed the downregulated expression of vascular endothelial growth factor and CD31 in diabetic mice (P < 0.05) and accelerated the wound angiogenesis process.

Conclusion: Roxadustat shows potential as a therapeutic drug by promoting re-epithelialization and angiogenesis to bring vigor to the impaired diabetic wound.