BackgroundLynch Syndrome (LS) is an autosomal dominant disease characterized by germline heterozygous mutations in DNA mismatch repair (MMR) genes. High-risk LS patients may proceed to colorectal cancer (CRC). However, the drivers or biomarkers of LS benign colon tissue approaching malignant CRC are not completely understood. This study aimed to understand the molecular and cellular changes during malignant transition in LS.MethodsSingle-cell RNA sequencing (scRNA-seq) was used to analyze paired fresh biopsy samples from 3 LS patients (carcinoma vs. para-carcinoma, labeled as LS-CA vs. LS-paraCA). Single-nuclear RNA sequencing (snRNA-seq) was used to analyze a frozen biopsy sample of a LS patient. Datasets of Healthy controls and patients diagnosed with sporadic CRC (without LS-related germline or somatic mutations; labeled as nonLS-CRC) were downloaded from the open source. Integrative computational analysis was performed to conclude potential drivers of the malignant transition. Immuno-histo-fluorescence staining (IHF) were also performed for validating the proposed three key markers.ResultsIn the single-cell atlas, we observed an increase of primitive cancer stem-cells with high expression of biomarkers CEACAM5, BACE2, GPRC5A and OLFM4 in the epithelium of the LS. Both infiltration of immune cells and pathways related to DNA repair biological activity in LS are dramatically increased in carcinoma compared to para-carcinoma. The mutation burden in LS is fundamentally elevated compared to that in healthy controls. Furthermore, T cell and macrophage-related tumor immunity in LS is readily mobilized in carcinomas compared to para-carcinoma.ConclusionsThis study provides single-cell transcriptomic resource using affected tissues from patients with Lynch Syndrome and describes an integrative profile covering the alterations of cancer stem cell markers, mutation burden, and tumor immunity during the malignant transition from latency state to Lynch Syndrome and to colorectal cancer at the single-cell level.

Purpose:To determine whether neutrophil extracellular trap (NET) predicts prognosis and response to neoadjuvant immunotherapy in gastric cancer (GC) and explore the associated mechanisms. Patients and Methods:Transcriptomic data from a GEO dataset (GSE62254) comprising 300 GC patients were analyzed. Patients were clustered based on 69 predefined NET-related genes (NRGs) summarized in previous studies, and clinical characteristics and immune cell infiltration between clusters were compared. An NRG signature was constructed. Retrospective clinical data and tissue samples from 243 surgically resected GC patients without neoadjuvant therapy and 49 patients receiving neoadjuvant chemotherapy combined with immunotherapy were collected. RNA sequencing, immunohistochemistry, and immunofluorescence were performed to assess NET density and its clinical relevance. Results:Two NET-related subtypes in GC (NT1 and NT2) with distinct clinical features and survival time were identified. A risk model based on five NRGs demonstrated that NT2 had lower risk scores, correlating with favorable outcomes. High NET density was associated with advanced TNM stage and short recurrence-free survival time in the surgery cohort. In the immunotherapy cohort, low pre-treatment NET density correlated with more T cells predicted superior response rates (45.8% vs. 16.0%, P = 0.032) and pathological complete response (29.2% vs. 4.0%, P = 0.023). Conclusion:Low NET density is linked to better prognosis and may identify patients with GC who could benefit from immunotherapy. These findings highlight the important role of NET in GC.

Malignant mesothelioma of the tunica vaginalis testis (MMTVT) is a rare malignancy originating from mesothelial cells of the testicular tunica vaginalis. Due to its insidious onset and local aggressiveness, most patients are diagnosed at an advanced stage. The treatment modalities for MMTVT primarily include surgery, chemotherapy, and radiotherapy. With the advent of immunotherapy, patient survival has further improved. To standardize and homogenize the diagnosis and treatment of MMTVT in China, the Chinese Alliance of Research for Mesothelioma, combining existing research evidence and nationwide expert opinions, has formulated a national expert consensus on the clinical diagnosis and treatment of MMTVT. This consensus encompasses epidemiology, diagnosis, treatment, prognosis, and follow-up.

BACKGROUND:The tumour immune microenvironment (TIME), particularly the presence and maturation of tertiary lymphoid structures (TLS), shapes antitumour immunity and therapy response. However, the role of mature TLS (mTLS) in rectal cancer (RC) and their modulation by neoadjuvant therapy (neoTx) remain unclear. METHODS:In this multicentre, retrospective study, we analysed patients with RC from two cohorts. Multi-omics profiling in patients with locally advanced rectal cancer (LARC) receiving no treatment (NT) included bulk RNA-seq (n = 123), immunohistochemistry and multiplex immunofluorescence (n = 161), scRNA-seq (n = 10) with paired scBCR-seq (n = 10). An independent neoTx cohort was used to assess treatment-induced immune changes, including bulk RNA-seq (n = 19) and immunohistochemistry (n = 125). FINDINGS:mTLS tumours were characterised by plasma cells and CD8+ T cells being located in close spatial proximity to each other. B cell-related signatures-including plasma cells, germinal centre B cells, and follicular B cells-as well as CD138, IgG, and IgA expression were elevated in mTLS tumours. scRNA-seq and scBCR-seq analyses further revealed that mTLS tumours harboured a greater abundance of plasma cells, broader clonal diversity, and a higher proportion of IgG+ and IgA+ plasma cells. High CD138 expression correlated with favourable survival. Post-neoTx tumours showed higher CD4+, CD8+, and CD45RO+ T cell densities and lower mTLS presence. Notably, B cell gene signatures and CD20+ cell density were enriched in responders to neoTx, despite no difference in TLS maturation. INTERPRETATION:mTLS are associated with enhanced B cell-mediated immune features and favourable prognosis. Post-neoTx is correlated with increased T cell infiltration but decreased TLS presence. The sustained B cell activation observed in non-responders raises the possibility that therapeutic strategies aimed at preserving or enhancing humoural immunity may benefit this patient subset. FUNDING:This study was supported by grants from the Natural Science Foundation of Beijing (7242034), New Technologies and Businesses of the PLAGH (5156ZE1X).

The incidence of HPV-related oral squamous cell carcinomas (OSCC)and oropharyngeal squamous cell carcinomas (OPSCC) has increased significantly in recent years, but the role of HPV and P16 in OSCC and OPSCC remains controversial. Here, we evaluate the prevalence and prognostic significance of HPV-DNA, HPV E6E7 and P16 protein expression in OSCC and OPSCC patients. Additionally, we explore the correlation between P16 protein expression and HPV infection in these cases. The results show that the HPV DNA infection rate was significantly higher in OPSCC at 16.7% compared to 3.6% in OSCC (P = 0.002), HPV DNA positive cases were more prevalent in poorly- differentiated cases (P = 0.009). HPV E6E7 positive cases (10.4%) were only detected in OPSCC. P16 (+++) was observed in 6 of 48 OPSCC cases (12.5%) and in 1 of 140 OSCC cases (0.7%). P16 (+++) was significantly higher in OPSCC than in OSCC (P = 0.003). However, in OPSCC, Using P16 (+++) as a marker for HPV DNA infection yielded a sensitivity of 62.5%, Kappa coefficient between HPV DNA and P16 (+++) was 0.67 (P <0.0001). HPV DNA infection and P16 (+++) were not linked to prognosis (DFS: P = 0.35, P = 0.51; OS: P = 0.99, P = 0.96), Moreover, In OSCC, Age, T, N, and clinical stages were correlated with prognosis. Our results indicate that P16 (+++) can act as a biomarker for HPV- related high-risk tumors in OPSCC, yet not in OSCC. Besides, Older patients, less favorable T, N, and clinical stages having a worse prognosis in OSCC. In those with OPSCC, only the younger age of patients was associated with a better prognosis.

Endometrial carcinoma (EC) is the most common malignancies of the female reproductive system in developed countries and areas. Ultrasound-guided and hysteroscopic samplings are commonly used to diagnose EC. However, clinicians question their diagnostic efficacy and the associated patient discomfort. DNA methylation is the widely studied epigenetic alteration in human tumors, and tumor screening and diagnosis. This review summarized common methods for collecting clinical samples for methylation testing. Furthermore, we analyzed the diagnostic evaluation indices of different methylation marker assays in clinical diagnosis and discussed the challenges of methylation testing in the future application of EC diagnosis.

The pan-immune-inflammation value reflects the systemic inflammatory response, and tumor-infiltrating lymphocytes indicate a local immune response in rectal cancer. However, the association between systemic inflammatory response, as indicated by the pan-immune-inflammation value, and local immune responses in rectal cancer remains unclear. This study analyzed 915 treatment-naïve rectal cancer patients from the Peking Union Medical College Hospital and PLA General Hospital (PLAGH) cohorts who underwent radical surgery to investigate the relationship between the pan-immune-inflammation value and immune responses. Lower pan-immune-inflammation value was significantly associated with improved disease-free survival and cancer-specific survival. Multivariate Cox regression models identified the pan-immune-inflammation value as an independent prognostic factor. In the PLAGH cohort, patients with low pan-immune-inflammation values had higher immune cell levels, activated immune pathways, and increased expression of immune checkpoint genes according to RNA sequencing. Hematoxylin and eosin staining and immunohistochemical analysis revealed that lower pan-immune-inflammation value was associated with higher tumor-infiltrating lymphocyte density, more mature tertiary lymphoid structures, increased CD8+ T cells, and elevated human lymphocyte antigen class I expression. Conversely, patients with high pan-immune-inflammation values exhibited pathways linked to tumor progression, such as angiogenesis, epithelial-mesenchymal transition, hypoxia, KRAS signaling, and TGF-ß signaling. Among patients receiving anti-PD-1 therapy, responders had low pre- and post-treatment pan-immune-inflammation values. The pan-immune-inflammation value is a reliable marker associated with distinct immune microenvironment characteristics and can effectively predict disease-free survival, cancer-specific survival, and response to immunotherapy.

Cullin-RING ubiquitin ligase 4 (CRL4) is closely correlated with the incidence and progression of ovarian cancer. DDB1- and CUL4-associated factor 13 (DCAF13), a substrate-recognition protein in the CRL4 E3 ubiquitin ligase complex, is involved in the occurrence and development of ovarian cancer. However, its precise function and the underlying molecular mechanism in this disease remain unclear. In this study, we confirmed that DCAF13 is highly expressed in human ovarian cancer and its expression is negatively correlated with the overall survival rate of patients with ovarian cancer. We then used CRISPR/Cas9 to knockout DCAF13 and found that its deletion significantly inhibited the proliferation, colony formation, and migration of human ovarian cancer cells. In addition, DCAF13 deficiency inhibited tumor proliferation in nude mice. Mechanistically, CRL4-DCAF13 targeted Fraser extracellular matrix complex subunit 1 (FRAS1) for polyubiquitination and proteasomal degradation. FRAS1 influenced the proliferation and migration of ovarian cancer cell through induction of the focal adhesion kinase (FAK) signaling pathway. These findings collectively show that DCAF13 is an important oncogene that promotes tumorigenesis in ovarian cancer cells by mediating FRAS1/FAK signaling. Our findings provide a foundation for the development of targeted therapeutics for ovarian cancer.

The BRAF gene is an important signaling molecule in human cells that is involved in the regulation of cell growth, differentiation, and survival. When the BRAF gene mutates, it can lead to abnormal activation of the signaling pathway, which promotes cell proliferation, inhibits cell apoptosis, and ultimately contributes to the occurrence and development of cancer. BRAF mutations are widely present in various cancers, including malignant melanoma, thyroid cancer, colorectal cancer, non-small cell lung cancer, and hairy cell leukemia, among others. BRAF is an important target for the treatment of various solid tumors, and targeted combination therapies, represented by BRAF inhibitors, have become one of the main treatment modalities for a variety of BRAF-mutation-positive solid tumors. Dabrafenib plus trametinib, as the first tumor-agnostic therapy, has been approved by the US Food and Drug Administration for the treatment of adult and pediatric patients aged 6 years and older harboring a BRAF V600E mutation with unresectable or metastatic solid tumors that have progressed following prior treatment and who have no satisfactory alternative treatment options. This is also the first time a BRAF/MEK inhibitor combination has been approved for use in pediatric patients. As research into the diagnosis and treatment of BRAF mutations advances, standardizing the detection of BRAF mutations and the clinical application of BRAF inhibitors becomes increasingly important. Therefore, we have established a universal and systematic strategy for diagnosing and treating solid tumors with BRAF mutations. In this expert consensus, we (1) summarize the epidemiology and clinical characteristics of BRAF mutations in different solid tumors, (2) provide recommendations for the selection of genetic testing methods and platforms, and (3) establish a universal strategy for the diagnosis and treatment of patients with solid tumors harboring BRAF mutations.

Background: Cold inducible RNA-binding protein (CIRP) is an important danger-associated molecular pattern involved in tissue-specific and systemic inflammation and is also regarded as a potential regulator of Alzheimer’s disease (AD). However, the precise roles and mechanism of CIRP in the functional changes in astrocytes during the development of AD are still unknown. This study aimed to assess gene expression alterations in astrocytes after they overexpress CIRP (oe-CIRP) and to explore the relationship between abnormal CIRP expression and AD. Methods: We established astrocyte cell lines that stably expressed CIRP or control vectors using 3 different kinds of human glioma cell lines, namely, U87, U251 and H4, and analyzed the mRNA expression profiles of 3 pairs of cells via microarray. Then, the significantly differentially expressed mRNAs between the CIRP-overexpressing (ov-CIRP) group and the control group were identified by bioinformatics analysis and validated by quantitative real-time PCR (q-PCR) and western blotting (WB). Finally, the effect of CIRP overexpression in astrocytes on neurons was observed in a coculture system. Results: We identified 119 mRNAs with obvious fold changes between the ov-CIRP and control groups for all 3 pairs of human glioma cell lines. These mRNAs are associated with diseases such as asthma IgE, dehydroepiandrosterone, rheumatoid arthritis, autism spectrum disorder, AD and so on. The biological functional analysis indicated that urokinase plasminogen activator (uPA), a gene whose expression significantly decreased after CIRP overexpression, was closely associated with AD. The results from q-PCR and WB assays confirmed that overexpression of CIRP significantly inhibited uPA at both the mRNA and protein levels in U87, U251 and H4 cells. Moreover, compared with those cocultured with control astrocytes, SH-SY5Y cells cocultured with CIRP-overexpressing astrocytes exhibited a significant increase in the expression of amyloid-β (Aβ)1-42 and the hyperphosphorylated microtubule-associated protein tau (Tau). Conclusion: CIRP overexpression inhibited the expression of uPA in human astrocytes, which promoted the expression of Aβ1‒42 and the phosphorylation of tau in neurons, thus increasing the risk of AD. These results suggest that the overexpression of CIRP in astrocytes contributes to the development of AD.

阑尾杯状细胞腺癌少见，本文报道1例以卵巢肿瘤为首发症状的转移性阑尾杯状细胞腺癌。患者女，67岁。腹痛2个月，CT示盆腔巨大囊实性肿物，考虑附件来源。术中送检左卵巢肿物，镜下观察大片坏死物中见印戒样/杯状细胞呈片状、筛状、腺管样或单个细胞排列。冷冻病理考虑转移性腺癌，建议临床检查消化道。术中探查发现阑尾肿物。遂行全子宫、双附件+右半结肠切除术。镜下观察少许阑尾肿瘤组织学形态与卵巢肿物一致，其他区域肿瘤细胞似神经内分泌细胞呈巢团、缎带状排列。卵巢和阑尾肿瘤均表达SATB2、CDX2，阑尾肿瘤局灶表达突触素。患者术后接受化疗。随访10个月，未见复发。.

患儿 男，出生48 h因“腹胀、腹股沟区皮肤瘀斑33 h”就诊。患儿出生15 h出现腹胀、双侧腹股沟瘀斑，伴贫血及血小板降低，腹部CT提示脾肿大、脾内多发低密度区，腹腔穿刺为血性腹腔积液。手术中见脾脏多个不均质充血区，行脾脏部分切除术并保留约20%脾脏，术后病理提示为卡波西样血管内皮瘤。术后腹腔出血停止，血红蛋白及血小板恢复正常。随访13个月患儿生长发育良好，未出现感染、腹胀及出血表现。

Histopathology image analysis plays a crucial role in cancer diagnosis. However, training a clinically applicable segmentation algorithm requires pathologists to engage in labour-intensive labelling. In contrast, weakly supervised learning methods, which only require coarse-grained labels at the image level, can significantly reduce the labeling efforts. Unfortunately, while these methods perform reasonably well in slide-level prediction, their ability to locate cancerous regions, which is essential for many clinical applications, remains unsatisfactory. Previously, we proposed CAMEL, which achieves comparable results to those of fully supervised baselines in pixel-level segmentation. However, CAMEL requires 1,280x1,280 image-level binary annotations for positive WSIs. Here, we present CAMEL2, by introducing a threshold of the cancerous ratio for positive bags, it allows us to better utilize the information, consequently enabling us to scale up the image-level setting from 1,280x1,280 to 5,120x5,120 while maintaining the accuracy. Our results with various datasets, demonstrate that CAMEL2, with the help of 5,120x5,120 image-level binary annotations, which are easy to annotate, achieves comparable performance to that of a fully supervised baseline in both instance- and slide-level classifications.

BackgroundTransformation of endometriosis to malignancy is a rare occurrence. Clear cell ovarian cancer and endometrioid ovarian cancer are the two histotypes most consistently linked to endometriosis. The exact pathways leading to malignant transformation of endometriosis remain elusive.Case presentationA 41-year-old woman presented to our hospital with a ten days history of abdominal pain which was not responsive to medication. Pathological examination revealed an unexpected finding of bilateral endometriosis associated with distinct malignancies: a clear cell carcinoma in the right ovary and a well-differentiated endometrioid carcinoma in the left ovary. Molecular analysis indicated a shared somatic driver mutation in ING1 in the eutopic endometrium and the bilateral ovaries while simultaneously exhibiting specific genetic alterations unique to each carcinoma. Notably, several common mutation sites were also identified, including previously reported common oncogenes (KRAS, PIK3CA, ARID1A). This finding prompts the hypothesis of a possible monoclonal origin of the two tumours.ConclusionThis case represents an exceedingly rare occurrence of two different histotypes of ovarian endometriosis-associated cancer manifesting simultaneously in bilateral ovaries. Based on genetic analysis, we hypothesize that these malignancies may have a monoclonal origin, providing insights into understanding the different biological mechanisms underlying carcinogenesis.

Cell senescence deters the activation of various oncogenes. Induction of senescence is, therefore, a potentially effective strategy to interfere with vital processes in tumor cells. Sphingosine-1-phosphate receptor 1 (S1PR1) has been implicated in various cancer types, including ovarian cancer. The mechanism by which S1PR1 regulates ovarian cancer cell senescence is currently elusive. In this study, we demonstrate that S1PR1 was highly expressed in human ovarian cancer tissues and cell lines. S1PR1 deletion inhibited the proliferation and migration of ovarian cancer cells. S1PR1 deletion promoted ovarian cancer cell senescence and sensitized ovarian cancer cells to cisplatin chemotherapy. Exposure of ovarian cancer cells to sphingosine-1-phosphate (S1P) increased the expression of 3-phosphatidylinositol-dependent protein kinase 1 (PDK1), decreased the expression of large tumor suppressor 1/2 (LATS1/2), and induced phosphorylation of Yes-associated protein (p-YAP). Opposite results were obtained in S1PR1 knockout cells following pharmacological inhibition. After silencing LATS1/2 in S1PR1-deficient ovarian cancer cells, senescence was suppressed and S1PR1 expression was increased concomitantly with YAP expression. Transcriptional regulation of S1PR1 by YAP was confirmed by chromatin immunoprecipitation. Accordingly, the S1PR1-PDK1-LATS1/2-YAP pathway regulates ovarian cancer cell senescence and does so through a YAP-mediated feedback loop. S1PR1 constitutes a druggable target for the induction of senescence in ovarian cancer cells. Pharmacological intervention in the S1PR1-PDK1-LATS1/2-YAP signaling axis may augment the efficacy of standard chemotherapy.

Ovarian cancer is a highly lethal form of cancer in females, largely due to extensive metastases that often accompany the initial diagnosis. Exosomes are microvesicles size from 30 to 100nm, which can be secreted by most cells. These special extracellular vesicles play a vital role in the metastasis of ovarian cancer. In this study, we conducted a comprehensive review of current research pertaining to the role of exosomes in ovarian cancer, utilizing the PubMed® and Web of Science databases. Our review highlights the progress in elucidating the mechanisms by which exosomes facilitate ovarian cancer progression. Additionally, we discuss the potential of exosomes as a novel therapeutic target for ovarian cancer treatment. Overall, our review provides valuable insights into the current state of research on exosomes in ovarian cancer therapy.

尿路上皮病变主要包括炎症和肿瘤.尿路上皮发生癌变后,部分肿瘤细胞会随尿液排出,因此,尿脱落细胞学检查已成为尿路上皮癌(urothelial carcinoma,UC)诊断和术后随访的主要方法之一.UC细胞依赖有氧糖酵解,作为维持其无限生长和增殖的主要能量来源,称为瓦博格(Warburg)效应.

畸胎瘤是妇科常见的生殖细胞肿瘤,好发于卵巢,其中95％为成熟性囊性畸胎瘤[1].性腺外畸胎瘤占生殖细胞肿瘤的2％～5％,较常发生于近中线部位.发生于宫颈的畸胎瘤罕见,其发生机制并不清楚.本研究报道2例原发于宫颈的成熟性畸胎瘤,并结合近年来国内外相关文献对宫颈原发性畸胎瘤的临床病理特点进行讨论.