Cytojournal最新文献

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.

Objective: Breast fine-needle aspiration cytology (FNAC) is a rapid, cost-effective, and minimally invasive diagnostic procedure. The diagnosis of a proliferative breast disease with atypia (PBDA) is established based on the presence of areas with disordered cellular arrangement and mildly discerned cytological features. We have aimed to explore the cytohistological correlation of PBDA on FNAC.

Material and methods: A review of the hospital database was undertaken to retrieve cases of breast FNAC diagnosed as PBDA between January 2011 and September 2020.

Results: A total of 3125 breast FNAC specimens were examined, and 107 (3.4%) of them received the diagnosis of PBDA. A total of 68 PBDA cases were included in this cytohistological evaluation. The risk of malignancy was 44%. Except for one case, all of the invasive or microinvasive carcinomas were grade 1 or 2 malignancies according to the Nottingham grading system of breast cancers. The result of a repeat FNAC of the case with a poorly differentiated invasive breast cancer was reported as a high-grade malignancy. A statistically significant correlation was observed between older age and malignant outcome (P < 0.001).

Conclusion: This is one of the largest datasets of cases with PBDA. Based on the advanced age of the patient, and relevant clinical and radiological information, cytopathological diagnosis of PBDA may prompt the clinician to take further action.

Objective: Hepatocellular carcinoma (HCC) represents a primary liver tumor characterized by rapid disease progression and unfavorable clinical outcomes. Most patients with HCC are identified in advanced stage, where targeted therapies are considered an effective treatment method for advanced disease. The tyrosine kinase inhibitor (TKI) donafenib has shown efficacy in managing HCC. However, drug resistance often occurs after treatment with donafenib, which limits its widespread clinical application. Thus, this study aims to identify small-molecule TKIs that can enhance the sensitivity of HCC to donafenib.

Material and methods: The HCC cells HepG2 and SNU449 were treated with five drugs, namely, dimethyl sulfoxide, AZ-628, SU-5402, TG-101209, and SPP-86, combined with donafenib to determine half-maximal inhibitory concentration values. RNA sequencing data obtained from The Cancer Genome Atlas (TCGA) were analyzed using Differential Expression analysis for Sequence data 2 (DESeq2)/limma and Gene Set Enrichment Analysis (GSEA). The effects of AZ-628 on proliferation, viability, apoptosis, and migration were assessed. The expression level of early growth response gene 1 (EGR1) was measured through Western blotting/quantitative polymerase chain reaction and silenced by cell transfection. Donafenib-resistant HepG2 cells negative control shRNA (shNC)/shRNA targeting EGR1 (shEGR1) were treated with AZ-628 combined with donafenib. Ferrous ion (Fe²⁺) and reactive oxygen species levels were measured after Erastin/RSL3 induction. The synergy between AZ-628 and donafenib was analyzed using Combenefit2. In vivo, tumor growth, and Ki67 expression were evaluated in nude mice treated with DMSO, AZ-628, donafenib, or their combination.

Results: This study showed that AZ-628 reduced donafenib resistance in HCC by targeting the tyrosine kinase (TK) pathway. Cell counting kit-8 and colony formation assay validated that AZ-628 significantly improved the sensitivity of HCC cells to donafenib (P < 0.0001). Rescue experiments showed that AZ-628 regulated HCC cell proliferation and drug resistance through EGR1 (P < 0.001). In addition, AZ-628 was found to affect donafenib resistance in HCC by regulating epithelial-mesenchymal transition, apoptosis, and ferroptosis (P < 0.0001). In vivo experiments demonstrated a combined anti-tumor efficacy of AZ-628 and donafenib in HCC models (P < 0.0001).

Conclusion: The findings of this study reveal a new combination therapy targeting the TK pathway for the treatment of HCC and provide a theoretical foundation for addressing donafenib resistance.

Objective: The function of tumor protein p53-inducible nuclear protein 2 (TP53INP2) in numerous cancers has been elucidated, but its role across the development of spinal cord glioma (SCG) remains largely unexplored. This study aims to explore the anti-cancer effect of TP53INP2/nuclear factor-kappa B (NF-kB)/nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway in SCG.

Material and methods: Quantitative real-time polymerase chain reaction was employed to determine TP53INP2 messenger RNA expression in vitro. Western blot analysis was conducted to detect TP53INP2, epithelial-tomesenchymal transition (EMT) marker, and NF-kB/Nrf2/HO-1 pathway protein. The proliferative potentials of glioma cells were assessed by 5-ethynyl-2'-deoxyuridine, colony formation, and cell counting kit-8 assays. Transwell assays were used to evaluate migratory and invasive capacities. Apoptotic cells and reactive oxygen species were analyzed using flow cytometer. Enzyme-linked immunosorbent assay was performed to measure superoxide dismutase and glutathione peroxidase levels. A tumor xenograft model in mouse was established.

Results: High expression of TP53INP2 was observed in glioma cells. TP53INP2 depletion significantly inhibited tumor growth, metastasis, EMT, and oxidative stress and increased the apoptosis rate and number of immune cells. The silenced TP53INP2 hampered the activation of NF-kB and promoted the activation of the Nrf2/HO-1 pathway.

Conclusion: This work highlights the therapeutic potential of TP53INP2/NF-kB/Nrf2/HO-1 axis in SCG.