Cytojournal最新文献

Objective: Ras p21 protein activator 1 (RASA1) plays a crucial role in the placenta. However, its effects and mechanisms of RASA1 on trophoblast function in preeclampsia (PE) are unclear. This study aims to investigate the relationship between the regulation of the Ras/mitogen-activated protein kinase (MAPK) pathway by RASA1 and the function of trophoblast cells in PE.

Material and methods: Placental tissues were collected from patients with early-onset PE and late-onset PE and their gestational age-matched normal pregnancies. Western blot analysis and quantitative reverse transcription polymerase chain reaction were employed to assess RASA1 levels in placental tissues and trophoblast cells, as well as Ras activation and p38 MAPK phosphorylation levels in the Ras/MAPK pathway. Wound healing, cell counting kit-8, and Transwell migration and invasion assays and flow cytometry experiments were conducted to detect the proliferation, migration, invasion, and apoptosis capabilities of trophoblast cells.

Results: RASA1 was significantly overexpressed in the placental tissues of patients with PE (P < 0.05). In cells, it downregulated the activation of Ras and phosphorylation of p38 MAPK (P < 0.05) and reduced proliferation and inhibited migration and invasive capabilities (P < 0.05). Moreover, RASA1 increased the rate of apoptosis, promoted the protein expression levels of cleaved caspase3 and Bax, and inhibited the expression of Bcl2 in cells (P < 0.05). The P38/MAPK inhibitor SB203580 reversed the activation of the Ras/MAPK pathway and the effects on proliferation, migration, invasion, and apoptosis of cells induced by si-RASA1 (P < 0.05).

Conclusion: The activity of the Ras/MAPK pathway could be inhibited by high RASA1 expression, which suppresses cell invasion, migration, and proliferation and boosts apoptosis. The abnormal regulation of the RASA1-Ras/MAPK axis may be a key factor in the development of PE and, therefore, provides new ideas and clinically effective strategies for the diagnosis and treatment of this condition.

Objective: Patients with chronic kidney disease (CKD) exhibit increased vascular calcification (VC) risks, worsened by high-dose erythropoietin (EPO). While EPO treats anemia, its role in VC pathogenesis remains unclear. Ginsenoside Rb1 (Rb1), a Panax ginseng compound with anti-calcification properties, may counteract EPO-induced VC through the GATA binding protein 6 (GATA6)/bone morphogenetic protein 2 (BMP2)/Smad1/5/9 pathway. This article aims to explore whether Rb1 could counteract EPO-induced VC through the GATA6/BMP2/Smad1/5/9 pathway.

Material and methods: Adenine-induced CKD rats and b-glycerophosphate-treated vascular smooth muscle cells (VSMCs) received EPO ± Rb1. Calcification was assessed through von Kossa/alizarin red staining. Smooth muscle protein 22-a (SM22a)/a-Smooth muscle actin (a-SMA) expression was measured by immunofluorescence and real-time-quantitative polymerase chain reaction (RT-qPCR). GATA6/BMP2/Smad1/5/9 activation was analyzed using RT-qPCR/Western blot. Rb1-BMP2 interactions were tested through biotin pulldown, micro-thermophoresis, and Co-immunoprecipitation (Co-IP). GATA6 knockdown validated pathway roles.

Results: High-dose EPO significantly worsened CKD-associated calcification and VSMC calcification (P < 0.01), suppressed SM22a and a-SMA expression levels, and activated the GATA6/BMP2/Smad1/5/9 pathway (P < 0.01). GATA6 knockdown reduced EPO-exacerbated calcification and modulated BMP2/Smad1/5/9 signaling (P < 0.01). Rb1 increased SM22a and a-SMA expression levels and inhibited Smad 1/5/9 phosphorylation (P < 0.01), without affecting GATA6 or BMP2 expression (P > 0.05). Molecular docking and Co-IP experiments revealed that Rb1 binds directly to BMP2, blocking its interaction with bone morphogenetic protein receptor and inhibiting Smad 1/5/9 phosphorylation (P < 0.01).

Conclusion: Rb1 mitigates EPO-aggravated VC in CKD by disrupting BMP2/Smad1/5/9 signaling, positioning it as a promising molecular intervention strategy to reduce EPO-induced vascular toxicity.

Low-grade fetal adenocarcinoma (L-FLAC) is an exceedingly rare subtype of lung adenocarcinoma, accounting for merely 0.3% of all pulmonary adenocarcinomas. Its accurate intraoperative cytological diagnosis poses substantial challenges, particularly in distinguishing it from carcinoid tumors, which have overlapping morphological features but different therapeutic approaches. We present the case of a 47-year-old female non-smoker with an incidental 21-mm pulmonary nodule spanning the right upper and middle lobes. Intraoperative fine-needle aspiration cytology demonstrated features suggestive of an atypical carcinoid tumor, primarily due to the presence of planar atypical cells with neuroendocrine-like characteristics. Comprehensive cytological assessment revealed two distinct cellular patterns: loosely cohesive planar cells alongside densely nucleated, three-dimensional cohesive clusters with cribriform architecture resembling endometrial tissue. Most importantly, focal cell aggregates with ground-glass nuclei and intranuclear inclusions, corresponding to morule-like structures - a pathognomonic feature of L-FLAC - were identified during detailed examination. Histopathological evaluation confirmed L-FLAC, characterized by atypical glandular proliferation with clear cytoplasm, subnuclear vacuolation, and distinctive morule-like structures demonstrating strong nuclear b-catenin positivity. The patient underwent lobectomy with lymph node dissection and remained recurrence-free at the 2-year follow-up. This case highlights four critical cytomorphological features essential for accurate intraoperative diagnosis of L-FLAC: (1) recognition of dual cell populations (loose planar cells versus cohesive endometrial-like clusters), which contrasts with the monomorphic presentation of carcinoid tumors; (2) identification of ground-glass nuclei and intranuclear inclusions in morule-like structures, features absent in carcinoid tumors; (3) presence of cribriform patterns within cohesive clusters; and (4) awareness that neuroendocrine-like features can dominate the cytological presentation. Accurate distinction between these entities is crucial, as carcinoid tumors may be amenable to limited resection in selected cases, whereas adenocarcinomas generally warrant lobectomy with lymph node dissection. Cytopathologists should remain vigilant for the subtle but diagnostic features of L-FLAC, particularly ground-glass nuclei and intranuclear inclusions, which provide definitive evidence for differentiating this entity from carcinoid tumors in challenging intraoperative settings.

Objective: Ovarian cancer is a disease that seriously endangers the health and life safety of women. At present, no effective preventive and therapeutic measures are available. This study probed the impact of cell division cycle-associated 2 (CDCA2) on ovarian cancer development and cisplatin sensitivity, which provides a new research direction for the study of ovarian cancer.

Material and methods: The protein expression level of CDCA2 was tested by Western blot assay. Cell proliferation was evaluated by cell cloning formation assay and Celigo cell counting. Cell invasion and migration were assessed by Transwell assay. An experiment for nude mouse tumor formation was conducted to analyze the influence of CDCA2 knockdown on tumor growth in vivo. We treated CDCA2 knockdown cells with gradient cisplatin and measured cell viability using the cell counting kit-8 assay. Apoptosis and DNA damage induced by CDCA2 knockdown were investigated by flow cytometry and histone family member X (H2AX) phosphorylated on Ser 139 (γ-H2AX) immunofluorescence, respectively.

Results: CDCA2 expression was knocked down in A2780 and SKOV3 cells. After CDCA2 knockdown, cell proliferation, migration, and invasion ability decreased significantly, and tumor growth in vivo was also limited (P < 0.01). The phosphorylation levels of protein kinase B (AKT) and mechanistic target of rapamycin (mTOR) were reduced by CDCA2 knockdown (P < 0.01), but the effect was reversed by the AKT activator SC-79 (P < 0.01). Knockdown of CDCA2 increased the cisplatin sensitivity of ovarian cancer cells by enhancing apoptosis and DNA damage (P < 0.01).

Conclusion: CDCA2 knockdown inhibited the development of ovarian cancer through the AKT/mTOR pathway and enhanced cisplatin sensitivity. CDCA2 is a potential target to reverse cisplatin resistance in ovarian cancer. It can also be used as a new research direction for the development of ovarian cancer therapy.

Objective: The objectives of the study are to investigate the differential metabolic paradigms and distribution of regulatory T (Tregs) cells between primary prostate cancer (PCa) and lymph node (LN) metastases.

Material and methods: Single-cell RNA sequencing analysis of primary PCa and LN metastases was employed to reveal the immune infiltration, identify Treg cell clusters, and analyze their metabolic regulation. Immunohistochemical (IHC) for FOXP3 and cluster of differentiation antigen 45 was used to verify different distribution and infiltration of Treg cells.

Results: Immune cell infiltration was prominent around PCa cells, with Tregs significantly enriched in node-positive samples, suggesting an immunosuppressive microenvironment. Three Treg subsets were identified: Inhibitory Tregs, effector Tregs, and double-positive Tregs, each exhibiting distinct metabolic profiles. IHC confirmed higher Treg infiltration in LN metastases compared to primary tumors, particularly within tumor stroma.

Conclusion: Tregs promote lymphatic metastasis in PCa through metabolic reprogramming, with their infiltration levels serving as a potential biomarker for metastatic risk.

Objective: Febrile seizures (FS) are common in pediatric epilepsy, but their precise etiology remains unclear. Although cytokines such as interleukin (IL)-4, IL-1, and IL-6 are known to influence FS fever responses, their specific role is not fully understood. This study aimed to clarify the correlation between IL-4 levels and febrile convulsions, exploring molecular mechanisms through bioinformatics and animal experiments to enhance the understanding of FS pathogenesis.

Material and methods: With the GSE28674 dataset, the K-means clustering algorithm was used to select key genes that regulate IL-4 during feature selection. An animal model of FS was developed, and in vivo experiments were conducted using enzyme-linked immunosorbent assay, flow cytometry, quantitative polymerase chain reaction (qPCR), Western blot, and immunofluorescence for validation.

Results: In this study, bioinformatics analysis and K-means clustering identified proto-oncogene (Jun), protooncogene (Fos), and Early growth response-1 (Egr1) as upstream regulators of IL-4. Dual-luciferase reporter assays confirmed that these transcription factors could activate the IL-4 promoter. qPCR and Western blot analyses showed that the messenger RNA (mRNA) and protein expression levels of Jun, Fos, Egr1, and IL-4 in the FS group were significantly higher than those in the normal control (NC) group (P < 0.05). In addition, immunohistochemical analysis demonstrated that the expression levels of these proteins in the FS group were significantly higher than those in the NC group (P < 0.001). The study also explored the impact of the IL-4 receptor blocker dupilumab on FS, revealing that the dupilumab group exhibited significantly reduced seizure latency (P < 0.001) and significantly increased seizure duration and Racine scores versus the FS group (P < 0.01). Furthermore, dupilumab significantly decreased the expression of IL-1β, tumor necrosis factor α, and IL-6 in serum (P < 0.001), as well as heat shock protein 70 mRNA and protein expression, glial fibrillary acidic protein, cysteine protease-3, and Bcl-2-associated X protein/B-cell lymphoma 2 in hippocampal tissue (P < 0.001). Flow cytometry analysis indicated an increased T helper cell type 1 (Th1)/T helper cell type 2 (Th2) cell ratio (P < 0.001) and increased apoptosis in the FS and dupilumab groups versus the NC group (P < 0.001), along with decreased cell cycle progression and proliferation ability (P < 0.001). Compared with the FS group, the dupilumab group exhibited a further increase in Th1/Th2 cell ratio and apoptosis (P < 0.001), along with a further decrease in cell cycle progression and proliferation ability (P < 0.001).

Conclusion: IL-4 and its upstream transcription factors Jun, Fos, and Egr1 may be associated with FS occurrence and development. Moreover, d

Primary effusion lymphoma (PEL) is a rare kind of extranodal non-Hodgkin large B-cell lymphoma that develops in the liquid phase in serous membrane-lined body cavities (perineum, pericardium, and pleura) without tumor masses. Kaposi sarcoma-associated human herpesvirus 8 (HHV8) is essential in establishing a diagnosis of PEL. An 84-year-old male with a past medical history of testicular cancer in his 40s presented with a chief complaint of shortness of breath which was attributed to a left pleural effusion. The flow cytometry indicated 32% small T lymphocytes (cluster of differentiation [CD]4: CD8 = 4.0:1), 9.0% small B lymphocytes without surface light chain expression, and 16% unknown phenotypic large cells. The cell block demonstrated large atypical lymphocytes with irregular nuclear membranes and coarse chromatin. The cells exhibited prominent nucleoli and relatively basophilic, abundant amphophilic cytoplasm. Multinucleated and Reed-Sternberg-like cells were also seen. We performed a panel of immunomarkers including HHV8 and ALK1. The tumor cells were positive for CD45, CD20, and HHV8 and negative for all other markers. Based on morphologic and immunophenotypical features, a diagnosis of PEL is rendered. Positron emission tomography/computed tomography showed an absence of fluorodeoxyglucose uptake in the lymph nodes and the spleen. Given his age, the patient started on treatment with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone at reduced doses (R-miniCHOP) and had a complete response. To our knowledge, there are no other reported cases of complete remission following an attenuated R-miniCHOP protocol in this clinical scenario.