Through in vitro and in vivo experiments, combined with network pharmacology and molecular docking techniques, this study investigated the mechanism of action of osthole in the treatment of colorectal cancer(CRC). The relevant targets of osthole and CRC were retrieved from the SwissTargetPrediction and SuperPred in drug databases, as well as GeneCards and OMIM in disease databases. Protein-protein interaction(PPI) networks were constructed using the STRING database and Cytoscape 3.8.0 software, and core targets were screened. Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were performed on common targets. Molecular docking validation of core targets with osthole was conducted using AutoDock Vina software. HCT116 cells were treated with different concentrations of osthole, and cell proliferation was detected using the CCK-8 assay and the clonogenic assay. Cell migration ability was assessed using Transwell assay. Western blot and RT-qPCR were performed to detect the expression of caspase-3(CASP3), hypoxia-inducible factor 1 alpha(HIF1A), nuclear factor kappa B subunit 1(NFKB1), glycogen synthase kinase-3 beta(GSK3B), phosphorylated-GSK3B(p-GSK3B), protein kinase B(Akt), phosphorylated-Akt(p-Akt), mammalian target of rapamycin(mTOR), and phosphorylated-mTOR(p-mTOR). A subcutaneous tumor model of HCT116 cells in nude mice was established, and the mice were randomly divided into the model group, low-dose osthole group(20 mg·kg~(-1)), medium-dose osthole group(40 mg·kg~(-1)), and high-dose osthole group(60 mg·kg~(-1)). After 18 days of administration, the growth of tumor xenografts was observed, and the size and weight of tumors were measured after excision. Hematoxylin-eosin(HE) staining was performed to observe the histological changes in tumors in each group. Network pharmacology analysis revealed that osthole treatment of CRC mainly involved 106 treatment targets and 113 treatment pathways, with key pathways including the PI3K/Akt signaling pathway and MAPK signaling pathway. Molecular docking results showed a strong correlation between osthole and core targets. In vitro studies demonstrated that osthole significantly inhibited the proliferation and migration ability of HCT116 cells. Western blot and RT-qPCR experiments showed that compared to those in the model group, the expression of NFKB1, HIF1A, p-Akt, p-mTOR, and GSK3B in the osthole-treated group was significantly decreased, while the expression of CASP3 and p-GSK3B(Ser9) was significantly increased. In vivo studies showed that compared to the model group, osthole-fed animals significantly reduced tumor weight and volume, inhibited tumor growth, and promoted tumor apoptosis, and the results showed a dose-dependent trend. The study suggested that osthole could inhibit the proliferation and migration of HCT116 cells in CRC, and its mechanism may be related to the regulation of the PI3K/Akt signaling pathway and the expression of core targets.

This study explored the generation site and regulation mechanism of reactive oxygen species(ROS) in the apoptosis of colorectal cancer cells induced by furanodienone(Fur). RKO cells were treated with 200 μmol·L~(-1) of Fur, and the changes in intracellular nicotinamide adenine dinucleotide phosphate oxidase(NOX) activity were detected by the NOX activity detection method. The control group, Fur group, diphenyleneiodonium(DPI) inhibitor group for general NOX, mitochondrial-targeted antioxidant(MitoTEMPO) group, Fur+DPI group, Fur+MitoTEMPO group, and H_2O_2 positive control group were set up. Intracellular ROS levels were detected by the ROS fluorescent staining method, and NOX1-NOX5 protein expressions were detected by Western blot. The NOX1-specific inhibitor ML171 and NOX4-specific inhibitor(GLX351322) were further introduced, and the cell activity was determined by cell counting kit-8(CCK-8) assay. The effects of ROS level change on the protein expressions of NOX4 and peroxiredoxin 1(PRDX1) were measured by Western blot. BAY11-7082, which is an inhibitor of the inhibitor of nuclear factor κB protein α(IκBα), was used to explore the effect of the expression of phosphorylated nuclear factor κB(p-NF-κB) in the nucleus after the Fur treatment on the NOX4 protein level. The lentiviral plasmid and empty plasmid for PRDX1 gene silencing were constructed to transfect RKO cells, and stably transfected strains were screened. The impact of PRDX1 gene knockout on Fur-induced apoptosis was further analyzed using the flow cytometry assay. The findings demonstrate a considerable increase in mitochondrial ROS level in response to Fur treatment, with an increase in intracellular NOX activity. However, the mitochondrial ROS level is significantly reduced in the Fur+DPI group. The results from Western blot and CCK-8 analysis suggest that intracellular NOX1 and NOX4 protein expressions are elevated by Fur treatment, and GLX351322 effectively reverses the pro-apoptotic effect of Fur, while ML171 has a minimal impact on apoptosis rate. Meanwhile, Fur significantly boosts the level of p-NF-κB in the nucleus, whereas the protein levels of p-NF-κB and NOX4 are reduced after the BAY treatment. The regulation of Fur on NOX4 and PRDX1 protein expressions is negatively correlated. In the stably transfected cell strain with PRDX1 gene knockout, the apoptosis rate is considerably higher than that of the negative control group after Fur treatment. The above results indicate that Fur can induce the apoptosis of colorectal cancer cells by promoting the signal transduction of NF-κB in the nucleus and increasing the generation of mitochondrial ROS derived from NOX4 to inhibit the PRDX1 protein expression.

This study investigated the effects of Agrimoniae Herba-Coptidis Rhizoma(XHC-HL)-medicated serum on the proliferation, migration, invasion, and apoptosis of human colorectal cancer HT29 and HCT116 cells via the autophagy mediated by lysosome-associated membrane protein type 2A(LAMP2A). Bioinformatics analysis was conducted to explore the role of LAMP2A in the development and progression of colorectal cancer. Western blot(WB) was used to detect the expression of LAMP2A protein in colorectal cancer cell lines. Lentiviral transfection was utilized to construct LAMP2A knockdown in HT29 and overexpression in HCT116 colorectal cancer cell models. Real-time fluorescence quantitative polymerase chain reaction(real-time qPCR) was performed to assess transfection efficiency. HT29 and HCT116 cells were treated with different concentrations of XHC-HL-medicated serum. The cell counting kit-8(CCK-8) assay was used to detect cell proliferation and determine the optimal concentration and duration of medicated serum intervention. HT29 cells were divided into a normal control(NC) group, an XHC-HL(medicated serum treatment) group, and an XHC-HL+shLAMP2A(medicated serum treatment+LAMP2A knockdown) group. HCT116 cells were divided into a NC group, an XHC-HL group, and an XHC-HL+LAMP2A(medicated serum treatment+LAMP2A overexpression) group. CCK-8 was used to measure cell viability. Colony formation assay was employed to assess cell proliferation ability. Scratch and Transwell migration assays were conducted to evaluate cell migration ability, and Transwell invasion assay was used to detect cell invasion ability. Flow cytometry was adopted to determine apoptosis rates. WB and real-time qPCR were employed to detect the effect of XHC-HL on the protein and mRNA expression of LAMP2A, heat shock cognate protein 70(HSC70), heat shock protein 90(HSP90), and glyceraldehyde-3-phosphate dehydrogenase(GAPDH) in colorectal cancer cells. Differential expression analysis revealed that LAMP2A expression was significantly higher in colorectal cancer patients compared to that in normal controls. Survival analysis indicated that the key molecule of chaperone-mediated autophagy(CMA), LAMP2A, was closely associated with colorectal cancer progression. Gene set enrichment analysis showed that patients with high LAMP2A expression significantly upregulated tumor progression-related signaling pathways such as angiogenesis and immune suppression. Immune infiltration analysis found that patients with high LAMP2A expression had fewer CD8 T cell infiltrations in their tumor microenvironment. XHC-HL-medicated serum inhibited the viability of HT29 and HCT116 cells, with the optimal intervention concentration and duration being 20% and 48 hours, respectively. Compared to the NC group, XHC-HL inhibited the proliferation, migration, and invasion of HT29 and HCT116 cells, and induced apoptosis. The medicated serum treatment with LAMP2A knockdown further inhibited colorectal cancer cell proliferation, invasion, and migration, and promoted apoptosis, whereas overexpression of LAMP2A reversed the inhibitory effects of the medicated serum on proliferation, migration, and invasion, and reduced apoptosis rates. XHC-HL-medicated serum inhibited CMA by upregulating the protein and mRNA expression of LAMP2A, HSC70, and HSP90 and downregulating substrate protein GAPDH expression via the autophagy mediated by LAMP2A. In conclusion, XHC-HL-medicated serum inhibits the proliferation, migration, and invasion of colorectal cancer cells and induces apoptosis by downregulating the expression of the key CMA molecule LAMP2A and inhibiting CMA activity.

Based on the signaling pathway of phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR), pathway-related phosphatase and tensin homolog(PTEN), B-cell lymphoma-2(Bcl-2), and Bcl-2-associated X protein(BAX), the mechanism of Rose roxburghii polysaccharides in inhibiting proliferation and inducing apoptosis of prostate cancer DU145 cells was explored, as well as the antioxidant activity of R. roxburghii polysaccharides. Prostate cancer DU145 cells were treated with different concentrations of R. roxburghii polysaccharides. The effect of R. roxburghii polysaccharides on the proliferation of DU145 cells was detected by the CCK-8 method, and the effect of R. roxburghii polysaccharides on the migration of DU145 cells was detected by cell scratch test. In addition, a Transwell assay was conducted to detect the effect of R. roxburghii polysaccharides on the invasion ability of DU145 cells. The apoptosis of DU145 cells induced by R. roxburghii polysaccharides was detected by flow cytometry. Real-time PCR and Western blot were used to detect the effects of R. roxburghii polysaccharides on the mRNA and protein expression levels of PI3K, Akt, mTOR, PTEN, BAX, and Bcl-2 in DU145 cells. DPPH and ABTS assays were used to determine the antioxidant activity of R. roxburghii polysaccharides in vitro. The results showed that R. roxburghii polysaccharides inhibited the prolife-ration, migration, and invasion of DU145 cells. Flow cytometry analysis showed that compared with that of the control group, the apoptosis rate of DU145 cells in groups treated with R. roxburghii polysaccharides was increased with the increase in the concentration of R. roxburghii polysaccharides, and its inhibition was positively correlated with the concentration of R. roxburghii polysaccharides. R. roxburghii polysaccharides inhibited the PI3K/Akt/mTOR pathway and induced apoptosis of DU145 cells by up-regulating the protein and gene expression of PTEN and BAX and down-regulating the expression of Akt, PI3K, mTOR, and Bcl-2. R. roxburghii polysaccharides could scavenge ABTS and DPPH radicals to a certain extent, suggesting that R. roxburghii polysaccharides may induce the apoptosis of DU145 cells by scavenging intracellular ROS. R. roxburghii polysaccharides may inhibit the proliferation of DU145 cells and induce its apoptosis by inhibiting the PI3K/Akt/mTOR pathway and clearing intracellular ROS.

This study investigated the anti-gastric cancer activity and mechanism of Panacis Quinquefolii Radix(Panax quinquefolium L.), and preliminarily compared the in vivo anti-gastric cancer efficacy of American-imported(JK-AG) and domestically produced(Shandong) Panacis Quinquefolii Radix decoctions(SD-AG). Based on network pharmacology predictions, a LUC-MGC803 cell ectopic gastric cancer nude mouse model was established. Mice were administered JK-AG and SD-AG at 1 g·kg~(-1) via gavage for 21 consecutive days. The positive control group received intraperitoneal injections of 5-fluorouracil(5-FU) at 20 mg·kg~(-1) every other day. Tumor volume was measured during the experiment. At the end of the experiment, high-resolution ultrasound imaging was used to measure tumor size, and an in vivo imaging system was used to observe tumor fluorescence expression. Tumor tissues were excised, weighed, and subjected to pathological histological examination. The terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL) assay and TUNEL+caspase-1 fluorescence double staining were used to detect tissue apoptosis and caspase-1 expression. Immunohistochemistry(IHC), Western blot(WB), and quantitative real-time PCR(RT-qPCR) techniques were used to detect the expression of predicted key proteins and genes involved in apoptosis and pyroptosis in tumor tissues. Network pharmacology predictions indicated that Panacis Quinquefolii Radix had anti-gastric cancer activity, primarily due to its ginsenoside components. Apoptosis induced by tumor necrosis factor(TNF)-α activation and caspase-1 may be key molecular mechanisms of its anti-cancer effects. In in vivo anti-tumor studies, compared to the model group, JK-AG and SD-AG did not affect the body weight or organ indices of the mice, whereas 5-FU significantly reduced body weight(P<0.05) and increased lung and liver indices(P<0.05). The tumor inhibition rates of JK-AG and SD-AG were 29.76% and 27.97%, respectively, which were lower than that of 5-FU. However, both JK-AG and SD-AG significantly reduced tumor tissue fluorescence intensity and three-dimensional tumor volume(P<0.05 or P<0.01). HE staining results showed that the drug groups had larger areas of tumor tissue necrosis. TUNEL and TUNEL+caspase-1 fluorescence staining indicated that both groups induced tumor cell apoptosis and increased caspase-1 expression. Both JK-AG and SD-AG significantly increased the expression of cleaved-caspase-8 and cleaved-caspase-3 proteins in tumor tissues detected by IHC and WB, as well as the mRNA expression of TNF-α, caspase-8, and caspase-9 detected by PCR, while reducing B-cell lymphoma 2(Bcl-2) protein expression. IHC showed that SD-AG increased the expression of TNF-α, caspase-9, cleaved-caspase-9, and tumor necrosis factor receptor 1(TNFR1) proteins more significantly than JK-AG, and also increased the Bax/Bcl-2 ratio, whereas JK-AG increased Bax mRNA expression more significantly than SD-AG. Additionally, JK-AG and SD-AG significantly increased the expression of NLRP3, caspase-1, cleaved-caspase-1, Gasdermin D(GSDMD), cleaved-GSDMD, IL-18, Gasdermin E(GSDME), and GSDME-NT proteins and the mRNA expression of caspase-1 and GSDME. JK-AG increased cleaved-caspase-1 protein expression more significantly than SD-AG in WB analysis. At the same time, JK-AG increased the expression of GSDMD mRNA, which was significantly higher than that of SD-AG group. For GSDME-NT protein expression, SD-AG increased it more significantly than JK-AG. The above studies indicate that Panacis Quinquefolii Radix has anti-gastric cancer effects, and the efficacy of American and Shandong ginsengs is comparable. Their mechanisms are related to TNF-α-mediated extrinsic apoptosis pathways and the induction of pyroptosis. The sensitivity of their action targets varies between American and Shandong ginsengs.

Objective: To study the effects and mechanisms of activation of human lung fibroblasts (MRC-5) by exosomal RNA hsa _ circ _ 0006357 (circEZH2) derived from non-small cell lung cancer. Methods: Western blot was used to detect exosome molecular markers, reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and cell invasion assays to detect the effect of non-small cell lung cancer-derived exosomes on MRC-5 activation. A circRNA microarray analysis was performed in serum exosomes from patients with non-small cell lung cancer (collected at Ningbo University People's Hospital, September 2023), and levels of circEZH2 were measured in serum exosomes from non-small cell lung cancer by RT-qPCR analysis. The effects of circEZH2 on MRC-5 activation were explored using wound healing assays, Transwell assays, RT-qPCR, cellular immunofluorescence, and western blot. The regulatory effect of circEZH2 on miR-495-3p/TPD52 axis and NF-κB pathway through dual-luciferase assay, immunofluorescence, and western blot. Results: Exosomes derived from non-small cell lung cancer cells were shown to promote MRC-5 cell invasiveness, the number of cells invaded in co-culture with exosomes derived from normal human bronchial epithelial cells was (42±5), and the number of cells invaded in co-culture with exosomes derived from non-small cell lung cancer cells (SPC-A1, H1299, A549 cells) was (246±7), (89±4), (69±14), expression of markers of fibroblast activation (α-SMA, FAP), and cytokines (IL-6, IL-8, P＜0.05). CircEZH2 expression was significantly upregulated in the serum exosomes of non-small cell lung cancer (P＜0.01). Alternatively, co-culture of exosomes derived from non-small cell lung cancer cells with MRC-5 cells promoted circEZH2 expression (P＜0.05). Functionally, overexpression of circEZH2 promoted MRC-5 cell migration and invasion [the cell migration rates were (30.81±2.54)% and (60.29±8.34)%, respectively, and the cell invasion numbers were (48.00±13.58) and (115.00±9.50), respectively, P＜0.05]. RT-qPCR, western blot, and immunofluorescence assays demonstrated a significant increase in the expression of the pro-inflammatory genes IL-6 and IL-8 in MRC-5 cells as well as the activation markers α-SMA and FAP in fibroblasts (P＜0.05) following the expression of circEZH2. Mechanistically, circEZH2 may function as ceRNA to regulate miR-495-3p, promote the expression of TPD52, and activate the NF-κB pathway to promote the activation of MRC-5. Conclusion: Exosomal circEZH2, derived from non-small cell lung cancer, may promote the activation of fibroblasts by activating the NF-κB pathway through the miR-495-3p/TPD52 axis.

Breast cancer is the most common malignant tumor among women in the world. According to data from the World Health Organization, in 2020, there were approximately 2.26 million new breast cancer cases worldwide, accounting for 11.7% of all new cancer cases. In recent years, with the rapid development of molecular biology and gene detection technology, the research on advanced breast cancer continues to deepen, and the treatment methods are constantly enriched. Gene targeted therapy significantly prolongs the survival period of patients with advanced breast cancer, which is of great significance for molecular pathological diagnosis, targeted drug selection and treatment mode optimization of patients with advanced breast cancer. Based on the development of literature and clinical research, the consensus expert committee formulated the "Expert consensus on clinical application of next-generation sequencing in advanced breast cancer (2024 edition)". Compared with the "Chinese Expert Consensus on Hot Issues of Gene Testing for Advanced breast cancer (2021)", the 2024 consensus adopts new evidence-based medical evidence, aiming to provide more comprehensive gene testing information for patients with advanced breast cancer, so as to develop more accurate treatment strategies.

Exploring the clinical and pathological characteristics and prognostic factors of diffuse large B-cell lymphoma (DLBCL) patients with TP53 mutation. Data of 86 DLBCL patients with TP53 mutation treated with R-CHOP and 19 DLBCL patients with TP53 mutation treated with R-CHOP like regimen as first-line treatment at the Cancer Hospital of Chinese Academy of Medical Sciences (CAMS) and the Cancer Hospital of the CAMS in Shenzhen, China, from January 2006 to June 2023 were retrospectively analyzed. Multivariate Cox analysis was applied to assess the effects of the factors on survival. Among the 105 DLBCL patients with TP53 mutation, 56 were male (53.3%); the median age was 59 years. There were 54 cases with stage Ⅰ-Ⅱ and 51 cases with stage Ⅲ-Ⅳ diseases. The proportion of B-cell lymphoma 2 (BCL2) gene amplification was 9.5% (10/105). The complete response rate in the whole group of patients treated with the R-CHOP regimen was 28.6% (30/105). The median progression-free survival (PFS) was 10.1 (95%CI: 7.3-13.0) months, and the median overall survival (OS) was not reached. Stage Ⅲ-Ⅳ was a risk factor for OS (HR=2.80, 95%CI: 1.04-7.54), and elevated lactic dehydrogenase (LDH) was a risk factor for PFS (HR=2.86, 95%CI: 1.56-5.26) and OS (HR=2.90, 95%CI: 1.08-7.69). The median PFS was lower in patients with BCL2 amplification than in patients without amplification [4.0 (95%CI: 2.7-5.3) vs 11.3 (95%CI: 8.5-14.1) months, P=0.011]. Thirty-one of the 54 (57.4%) patients with stage Ⅰ-Ⅱ disease received combination therapy based on the R-CHOP protocol. In conclusion, stage Ⅲ-Ⅳ and elevated LDH were associated with poor prognosis in TP53 mutation DLBCL patients, and patients with BCL2 amplification had a poor prognosis. For TP53 mutation DLBCL patients with stage Ⅰ-Ⅱ disease, combination of therapeutic modalities based on the R-CHOP may improve the prognosis.

Malignant tumors are closely related to various genetic and environmental factors. Pathogenic germline gene mutations play a key role in the occurrence and development of some malignant tumors. Some germline mutations can increase the risk of malignant tumors. For example, those with homologous recombination repair gene BRCA1/2 mutations are prone to breast cancer, ovarian cancer, etc., and some germline mutations are associated with genetic syndromes. For instance, 80% of hereditary non-polyposis colon cancers are associated with mutations in mismatch repair genes such as MLH1 and MLH2. In addition, 70% of Li-Fraumeni syndrome patients have harbored germline TP53 mutations. With the development of next-generation sequencing technology, more and more germline gene mutations have been discovered recently, which is of great significance for the prevention, screening, and treatment of tumors. This article has provided a review for common germline mutations, detection methods, and advances in drug therapy.

Objective: To investigate the clinicopathological features, immunophenotype, molecular characteristics, and differential diagnosis of primary intracranial DICER1-mutant sarcoma in order to better understand this tumor type. Methods: A retrospective analysis was conducted on 7 cases of primary intracranial DICER1-mutant sarcoma diagnosed in the Department of Pathology, Sun Yat-sen University Cancer Center, Guangzhou, China between 2021 and 2023 using next-generation sequencing. At the same time, 10 gliosarcomas, 4 intracranial FET::CREB fusion-positive mesenchymal tumors, 4 malignant meningiomas, 3 malignant solitary fibrous tumors, 3 malignant peripheral nerve sheath tumors, 3 synovial sarcomas and 3 rhabdomyosarcomas (total 30 cases) were selected as control. Results: Among the 7 patients with primary intracranial DICER1-mutant sarcoma, 6 were male and 1 was female, aged 10-32 years (median, 23 years). The tissue morphology was predominantly spindle or pleomorphic sarcoma-like, with 6 cases exhibiting eosinophilic globules, and 3 cases showing rhabdomyoblastic or rhabdomyosarcoma-like cell differentiation. Immunohistochemistry revealed focal desmin expression in 3 cases (3/7), ATRX loss in 3 cases (3/7), and p53 mutant pattern in 4 cases (4/7). Additionally, 4 cases (4/7) showed focal or diffuse SALL4 expression, whereas the control cases (30 cases) did not exhibit SALL4 protein expression, suggesting that SALL4 may possess certain auxiliary diagnostic value. Next-generation sequencing confirmed that all 7 cases of primary intracranial DICER1-mutant sarcoma harbored mutations in the DICER1 gene, with 5 cases having the mutation site at p.E1813D. Until May 2024, all 7 patients were alive. Conclusions: Primary intracranial DICER1-mutant sarcoma is a rare tumor. Understanding its morphological characteristics, immunohistochemical and molecular markers and differential diagnosis is crucial to avoid misdiagnosis and to improve diagnostic accuracy of this tumor.

Objective: To investigate the clinicopathological and molecular genetic features of POLE mutant endometrioid carcinoma. Methods: Genetic test data of 230 cases of endometrial carcinoma that underwent surgical resection and molecular typing by next generation sequencing in the First Medical Center of Chinese PLA General Hospital from January 2021 to June 2023 were retrospectively analyzed. Seventeen cases of endometrioid carcinoma with POLE mutation were selected. Clinical and prognostic information was collected. The paraffin-embedded tissue and immunohistochemical sections were reviewed, and the gene detection data were analyzed. Results: In the 17 cases of endometrioid carcinoma with POLE mutations, 16 cases (16/230, 6.9%) had mutations at known pathogenic sites, and 1 case had a mutation site (S459Y) that had not been reported, which was inferred to be pathogenic based on clinical prognosis. The 17 patients aged from 48 to 79 years (median 56 years, mean 58 years). All cases had typical histological features of endometrioid carcinoma, including 7 cases (7/17) of poorly-differentiated, 4 cases (4/17) of moderately-differentiated and 6 cases (6/17) of well-differentiated. Squamous differentiation was noted, mucous differentiation was less commonly found and often accompanied by superficial muscle infiltration. The number of stromal lymphocyte infiltration was variable. Lymph-vascular embolus was found in 6 cases, and lymph node metastasis was only detected in 1 case. According to the FIGO staging system for endometrial cancer in 2023, all the cases were in FIGO stage ⅠAm-POLEmut except for one case in FIGO stage ⅢC1. There were 8 cases with genetic co-mutation, 5 cases with TP53 mutation (immunohistochemically subclonal expression pattern), 1 case with MSI-H, and 2 cases with both TP53 mutation and MSI-H. Five of 7 patients with POLE mutation (poorly-differentiated) received postoperative chemotherapy and/or radiotherapy, 4 patients received endocrine therapy, and 8 patients had no treatment after surgery. One of the stage ⅠAm-POLEmut tumor patients was found to have pelvic recurrence one year after surgery, and the other 16 patients were followed up for 10-38 months without recurrence or metastasis. Conclusions: POLE mutant endometrioid carcinoma may have different differentiation, and most patients have good prognosis. Correct interpretation of molecular results, accurate identification and classification are important for predicting prognosis and avoiding overtreatment. However, a small number of cases may have recurrence and metastasis, and therefore it is necessary to make a reasonable treatment plan based on the comprehensive judgment of other high risk factors.

Objective: To investigate the clinicopathological, molecular pathological features, and family genetic pedigree of small cell carcinoma of the ovary, hypercalcemic type (SCCOHT). Methods: A total of 16 cases of SCCOHT diagnosed in Obstetrics and Gynecology Hospital of Fudan University from January 2013 to January 2023 were collected. The clinicopathologic features, SMARCA4/2/B1 protein expression, outcomes and SMARCA4 gene detection were reported. A follow-up study was also carried out. Results: The average age at diagnosis was 28.7 years (range 17-38 years). The preoperative calcium level was evaluated in 3 of 6 patients. The tumor was unilateral in all 16 cases, ranged from 8 to 26 cm (average 15.8 cm) in the greatest dimension. Extraovarian spread was present in 7 cases. In 10 cases, the tumors were initially misinterpreted as other ovarian neoplasms. BRG1 and BRM expression by immunohistochemistry were all lost in detected cases, while INI1 exhibited retained nuclear expression. All BRM-negative SCCOHTs also lacked BRG1 protein,but retained INI1 expression. SCCOHTs were only focally positive for EMA, CKpan, Calretinin, SALL4, and diffusely positive for WT1. Two of nine cases exhibited mutation-type p53 immunoreactivity. Ki-67 index was 58% on an average. ER, PR, FOXL2, α-inhibin, chromogranin A and LCA were negative in all the cases. SMARCA4 sequencing was available in 8 cases of SCCOHT, which revealed a germline SMARCA4 mutation in one patient, and others carried somatic mutation. Furthermore, two daughters, mother and an aunt of a patient with germline mutation were reported to be SMARCA4 mutation carriers. Follow-up was available for 15 patients, and the 6-month, 1-year and 2-year survival rate was 65.8%, 45.1%, and 22.6%, respectively. For patients in FIGO stages Ⅱ+Ⅲ, 6-month, 1-year survival rate was 53.6% and 35.7% respectively, compared to 80% (6-month) and 60% (1-year) in patients of staged I (P=0.358). Conclusions: With dismal prognosis of SCCOHT, accurate diagnosis is necessary. The typical age distribution, a panel of various staining results, especially concomitant loss of BRG1 and BRM may be of diagnostic aid and can be used to distinguish SCCOHT from its histological mimics. After the diagnosis of SCCOHT, genetic testing and genetic counseling are recommended.

Objective: To investigate the relationship between the expression patterns of SOX2 and FOXG1 and the differentiation/development level of neural components in immature teratoma and to determine the clinical significance and potential application of this correlation in a clinical setting. Methods: We conducted a comprehensive whole transcriptome sequencing analysis to identify differentially expressed genes (DEGs) across various subtypes of ovarian germ cell tumors. Additionally, immunohistochemical staining of paraffin-embedded tissue sections was employed to assess the nuclear staining pattern of SOX2 and FOXG1 proteins within the tumor tissues. Results: The transcriptome sequencing data showed that transcription factors SOX2 and FOXG1 exhibited high levels of expression typically in immature teratoma and occupied a pivotal position within the protein-protein interaction network. Immunohistochemical staining revealed the absence of both SOX2 and FOXG1 protein expression in dysgerminoma and yolk sac tumor samples. In immature teratoma, immunohistochemical staining demonstrated diffuse expression of SOX2 and FOXG1 proteins within the inner layer of densely-arranged primitive neuroepithelial tubules. This pattern of expression suggested the presence of stem cell-like properties within these tumor cells. In the sparsely peripheral neurogliocytes, FOXG1 maintained a diffuse nuclear staining pattern resembling that of neuroepithelial cells, while SOX2 exhibited a scattered pattern of positive staining, hinting at a neural lineage differentiation potential. This spatial differential expression pattern of SOX2 and FOXG1 proteins in immature teratoma suggested that primitive neural components within these tumors often recapitulated the trajectory of neural formation and cortical development that was typically observed during embryogenesis. The primitive neural tube acted as the center that constantly moved from inside to outside, with a dynamic shift from the interior to the exterior, paralleled by the sequential differentiation of cell lineages from primitive neuroepithelial stem cells to radial glia, intermediate progenitor cells, and ultimately to precursor glia. Conclusions: This spatial expression pattern of SOX2 and FOXG1 proteins observed in immature teratoma mirrors the lineage differentiation and migration trajectories of primitive neuroepithelial components typically seen in embryonic neurogenesis and cortical development. In daily practice, the combined application of SOX2 and FOXG1 SOX2 and FOXG1 helps identify the primitive neuroepithelial components in immature teratoma, avoid misjudgment of similar morphologies, and thereby assist in the histological grading and clinical decision-making.