e17511 Background: Circulating tumor HPV DNA (ctHPV) is emerging as a promising biomarker for minimal residual disease (MRD) in cervical cancer. Prospective studies investigating the association of ctHPV at diagnosis with primary tumor size and lymph node metastasis (LNM), as well as its potential for longitudinal MRD monitoring, are limited. Establishing a rapid and clinically feasible method for ctHPV quantification could improve early risk stratification and guide precision management strategies. Methods: Newly diagnosed HPV16/18-positive cervical cancer patients are being prospectively enrolled, with a planned cohort of 83 patients. 76 patients have been analyzed to date. ctHPV levels at diagnosis were quantified using droplet digital PCR (ddPCR) and next-generation sequencing (NGS). The concordance between ddPCR and NGS was evaluated using correlation analysis and Bland–Altman plots. Associations of ctHPV with LNM status and trends according to primary tumor size were analyzed. Post-treatment ctHPV measurements are ongoing, and patients will continue longitudinal follow-up to evaluate ctHPV dynamics and their potential association with treatment response and prognosis. Results: Analyses presented here include 76 patients from the ongoing prospective cohort. ctHPV was detectable at diagnosis in all patients. ddPCR showed high concordance with NGS, with Bland–Altman analysis demonstrating minimal bias, supporting its use as a rapid and clinically feasible quantification method. Patients with LNM exhibited higher ctHPV levels at diagnosis compared with those without LNM, and ctHPV levels increased with greater primary tumor size. These findings highlight the potential of ctHPV to reflect tumor burden and LNM status at diagnosis. Conclusions: ctHPV at diagnosis is associated with primary tumor size and lymph node metastasis in cervical cancer. ddPCR provides a rapid and reliable method for ctHPV quantification. Ongoing and future longitudinal analyses will explore ctHPV dynamics during treatment and follow-up as a marker for MRD and prognosis, supporting its potential clinical utility in prospective patient management. Circulating tumor HPV DNA (ctHPV) levels measured by ddPCR and NGS in the prospective cohort, showing stratified by lymph node metastasis and primary tumor size. Analysis/cohort N ddPCR log10 (ctHPV + 1) (copies/ml) NGS log10 (ctHPV + 1) (copies/ml) Mean difference (95% LoA) P Value (ddPCR) P Value (NGS) ddPCR/NGS concordance 76 2.091 ± 1.19 2.27 ± 1.29 -0.18 (-0.72 – 0.36) # - - LNM positive 37 2.58 ± 1.24 2.80 ± 1.41 - 0.0003 0.0003 LNM negative 39 1.63 ± 0.94 1.77 ± 0.93 - Tumor <= 2 cm 9 1.25 ± 0.91 1.46 ± 0.89 - 0.0002 * 0.0005 * Tumor 2 – 4 cm 21 1.57 ± 0.86 1.71 ± 0.79 - Tumor >= 4 cm 46 2.49 ± 1.20 2.69 ± 1.38 - #: Bland–Altman analysis; *: Linear trend was evaluated by polynomial contrasts in one-way ANOVA.

Objective To investigate the predictive value of the cancer antigen-125 elimination rate constant K(KELIM)for treatment response and prognosis in patients with advanced high-grade serous ovarian cancer(HGSOC)undergoing neoadjuvant chemotherapy followed by interval debulking surgery(NACT-IDS),and to analyze the combined prognostic significance of KELIM and the mutation status of breast cancer susceptibility gene(BRCA).Methods A total of 106 patients with advanced HGSOC who had undergone NACT-IDS were retrospectively enrolled.The KELIM values during neoadjuvant chemotherapy were calculated,and patients were divided into high-and low-KELIM groups using a cutoff value of 1.0.Clinicopathological characteristics,R0 resection rates,and platinum sensitivity rates were compared between the two groups.Logistic regression analysis was performed to identify predictive factors for R0 resection,while Kaplan-Meier survival analysis and Cox proportional hazards regression were performed to evaluate factors associated with progression-free survival(PFS).Furthermore,the patients were stratified according to both KELIM and BRCA status to assess the risk of platinum-resistant recurrence in each subgroup.Results The R0 resection rate was higher in the KELIM≥1 group than in the KELIM<1 group(77.1%vs 55.2%),and the difference was statistically significant(P=0.024).Multivariate logistic regression analysis showed that KELIM was an independent predictor of R0 resection(odds ratio[OR]=2.922,95%CI:1.112-7.678).Survival analysis demonstrated longer PFS in the KELIM≥1 group compared with that in the KELIM<1 group(33.0 months vs 18.0 months),and the difference was statistically significant(P<0.001).Multivariate Cox regression analysis showed that KELIM≥1 was associated with a reduced risk of disease progression(hazard ratio[HR]=0.481,95%CI:0.280-0.826).Combined stratification analysis revealed that no platinum-resistant recurrence was observed in the subgroup with both KELIM≥1 and a BRCA-positive status(0/21).Compared with patients with KELIM<1 and a BRCA-negative status,this subgroup exhibited a lower risk of platinum-resistant recurrence(OR=0.053,95%CI:0.003-0.932,P=0.006).Conclusion KELIM is an effective dynamic biomarker for predicting surgical outcomes and PFS in patients undergoing NACT-IDS.Combined stratification by KELIM and BRCA status allows more precise identification of the patient population with both KELIM≥1 and BRCA-positive status,who have an extremely low risk of platinum-resistant recurrence,thereby providing an important basis for individualized treatment and risk stratification management in patients with advanced HGSOC.

High-grade serous ovarian carcinoma (HGSOC) remains the most lethal gynecologic malignancy, with platinum resistance posing the major therapeutic barrier. Malignant ascites, a hallmark of advanced HGSOC, correlates with chemoresistance and poor prognosis, yet the contribution of ascites circRNAs in this process remains obscure. Here, we identify ASCOR, a circRNA upregulated in ascites small extracellular vesicles (sEVs) from platinum-resistant HGSOC patients, predicts poor survival. ASCOR promotes platinum resistance in vitro and in vivo by enhancing cell viability, reducing apoptosis, and alleviating DNA damage, with effects also transferred via sEVs. Mechanistically, ASCOR binds RPA1 and facilitates its NUP153-dependent nuclear translocation, which recruits the RNA helicase DDX18, suppressing R-loop formation. Meanwhile, the ASCOR-RPA1-DDX18 axis activates PI3K/Akt signaling to promote platinum resistance. Targeting ASCOR with antisense oligonucleotides reverses chemoresistance in mouse models. Our study unveils an ascites circRNA-driven pathway underlying platinum resistance in HGSOC, suggesting targeting ASCOR is a promising strategy for HGSOC treatment.

BACKGROUND:Epithelial ovarian cancer (EOC) is characterized by a suppressive tumor immune microenvironment (TIME) that undermines therapeutic efficacy. Immunogenic cell death (ICD) is a promising immunotherapy strategy attributed to ameliorating the suppressive TIME. Cisplatin (CDDP), the first-line chemotherapy for EOC, is hindered by cumulative toxicity. Cucurbitacin I (CuI), a bioactive plant-derived compound, exhibits remarkable antitumor activity in preclinical models. However, its ICD-inducing capability and cisplatin-sensitizing potential in EOC remain uninvestigated. PURPOSE:This study aims to systematically elucidate the ICD-inducing capability of CuI and its potential to enhance CDDP's antitumor effects in EOC both in vitro and in vivo. METHOD:The inhibitory effects of CuI on EOC were evaluated at the cellular level. The half-maximal inhibitory concentrations (IC50) of CuI in EOC cell lines were determined via CCK-8 assay. CuI's effects on cell cycle, pyroptosis (Annexin V+/PI+), immune gene expression profile, and GSDME/cleaved-caspase-3 were assessed in ID8/OVCAR8 cells. Pyroptotic pathways were validated using GSDME siRNA and caspase-3 inhibitor (Z-DEVD-FMK). Following the Nomenclature Committee on Cell Death (NCCD) guidelines for ICD inducer validation, we systematically assessed damage-associated molecular patterns (DAMPs) release (CALR, ERp57, HMGB1, HSPs), dendritic cell (DC) maturation (flow cytometry analysis of murine bone marrow-derived dendritic cells (BMDCs) co-cultured with CuI-pretreated ID8 cells), and in vivo tumor vaccine potency. Intraperitoneal xenograft models evaluated CuI's antitumor efficacy and immune infiltration. Finally, CuI-CDDP synergy was tested in EOC cell lines and immunocompetent models; targets were predicted by bioinformatics (Connectivity Map and molecular docking) and validated by Surface Plasmon Resonance (SPR) to elucidate mechanistic interactions. RESULT:CuI induced caspase-3/GSDME-mediated pyroptosis in EOC cells, driving the release of ICD-related DAMPs, including surface-exposed CRT/ERp57, ATP, and HMGB1. Consequently, CuI-treated EOC cells promoted DC maturation, significantly increasing the proportion of CD80+CD86+ DCs in tumor-draining lymph nodes (from 16.07±1.02 % to 23.47±1.94 %). In ID8-luc intraperitoneal xenograft, CuI could effectively suppress ascites formation by 41.4 % and was associated with an approximately 2.8-fold increase in CD8+T cell infiltration. Furthermore, the combination of CuI and CDDP synergistically enhanced pyroptosis in vitro. This synergy translated to superior antitumor effects in the ID8 xenograft model, achieving a tumor growth inhibition (TGI) of 66.02 %. This potent efficacy was accompanied by enhanced CD8+T cell infiltration, a reduction in PD-1 expression on CD8+T cells, and an approximately 1.78-fold increase in the splenic effector memory CD8+T cell (CD44+CD62L-) population compared to the control. Mechanistically, CuI may enhance CDDP-induced intracellular DNA damage and reactive oxygen species accumulation by inhibiting EGFR activation. CONCLUSION:CuI is a novel ICD inducer that elicits caspase-3/GSDME-mediated pyroptosis and boosts antitumor immunity in EOC. Notably, it enhances the chemosensitivity of EOC to CDDP, offering a promising strategy for EOC treatment.

TMVP1446 is a novel peptide independently screened by our research team, which can target tumors and lymph node metastases in vivo due to its specific binding to VEGFR-3 protein. However, its poor stability in vivo greatly limits its clinical translation. Herein, through a series of modifications to protect the TMVP1446 peptide from protease degradation, TMVP1446/GS5, a novel 12-amino acid peptide that retains specific binding to VEGFR-3 and exhibits improved stability in vivo, was designed. The low-millimolar dissociation constant (KD) of VEGFR-3 binding, VEGFR-3-dependent binding to cells, and competitive binding with TMVP1446 confirmed the specific affinity of TMVP1446/GS5 to VEGFR-3 in vitro. TMVP1446/GS5 was conjugated to the near-infrared (NIR) dye Cy7 for visualization of tumor and lymph node metastases in vivo. Surprisingly, despite having nearly the same KD value as TMVP1446, TMVP1446/GS5-Cy7 exhibited much stronger fluorescence signals in vivo in tumor and lymph node metastases. This paradoxical result further confirmed that TMVP1446/GS5 showed increased in vivo metabolic stability relative to TMVP1446. Additionally, TMVP1446/GS5 exhibited good biocompatibility in vivo safety assessment experiments. In conclusion, the novel peptide TMVP1446/GS5, with a specific binding affinity to VEGFR-3 and improved stability compared to TMVP1446, has great potential in the clinical application of fluorescence imaging of tumors and lymph node metastases.

Cervical cancer (CC) is a major health threat to women, with immunotherapies targeting the programmed death receptor 1/programmed death ligand 1(PD-1/PD-L1) axis showing promise but encountering resistance in a significant patient population. This resistance has driven a critical quest to uncover the underlying mechanisms. This study uncovers a novel metabolic axis involving the nicotinamide adenine dinucleotide (NAD+) salvage pathway enzyme nicotinamide phosphoribosyltransferase (NAMPT) and the deacetylase Sirtuin 1 (SIRT1), which regulates PD-L1 expression and nuclear localization in CC. This axis may be a key factor contributing to the resistance observed in immunotherapy. This study reveals that PD-L1 overexpression in cancers is regulated by both transcriptional and post-transcriptional processes. Acetyl-proteomic analysis pinpoints SIRT1 as a central regulator in the deacetylation of histone H3 at lysines 27, which may influence PD-L1 subcellular distribution. This finding reveals the epigenetic control of immune checkpoint proteins by metabolic pathways, offering a new perspective on the regulation of PD-L1. The identification of the NAMPT/SIRT1 metabolic axis as a critical factor suggests that targeting this axis may enhance therapeutic responses.

Rationale: Pyroptosis, an emerging form of programmed cell death, facilitates the release of tumor antigens and inflammatory factors, which can be leveraged to enhance the efficacy of immune checkpoint blockade (ICB) therapy. However, achieving high-efficiency induction of pyroptosis in cancer cells while minimizing toxicity remains a significant challenge. Methods: In this study, we designed a tumor-targeting peptide TMTP1-modified nanostructured lipid carrier (referred to as TP-NLC) with high loading capacities for gambogic acid (GA) and indocyanine green (ICG). The TMTP1, identified by our research team for its tumor-targeting capabilities, was conjugated to the nanocarrier surface using "click chemistry" to improve the drug delivery efficiency to tumor tissues. The TP-NLC nanocarrier was thoroughly characterized with respect to its morphological attributes, photostability, tumor-targeting capabilities, ability to induce pyroptosis, reactive oxygen species (ROS)-responsive behavior, and anti-tumor efficacy both in vitro and in vivo. Results: GA encapsulated within the TP-NLC nanocarrier, induced pyroptosis in tumor cells, and enhanced the efficacy of ICG-induced pyroptosis under laser irradiation by disrupting intracellular antioxidant systems, realizing that the combination of GA and ICG synergistically induced caspase-3/GSDME-mediated pyroptosis in a ROS-dependent manner. Tumor cells of pyroptosis released cellular contents and tumor antigens, which subsequently promoted the maturation of dendritic cells (DCs), enhanced intratumoral infiltration of CD8+ T cells, initiated systemic antitumor immune response, and augmented the efficiency of PD-1 blockade against both primary and metastatic tumors. Conclusion: The combination of GA and ICG therapy utilizing the constructed nanocarriers presents an attractive therapeutic strategy to trigger pyroptosis and potentiate PD-1 blockade therapy for cervical cancer chemo-immunotherapy.

Cholangiocarcinoma (CCA) arises within the peritumoral bile microenvironment, yet microbial translocation from bile to intracholangiocarcinoma (IntraCCA) tissues remains poorly understood. Previous studies on bile microbiota alterations from biliary benign disease (BBD) to CCA have yielded inconsistent results, highlighting the need for cross-study analysis. We presented a comprehensive analysis of five cohorts (N = 266), including our newly established 16S rRNA gene profiling (n = 42), to elucidate these microbiota transitions. The concordance of bacteria between CCA bile and intraCCA tissue, represented by Enterococcus and Staphylococcus, suggested microbiota migration from bile to intratumoral tissues. A computational random forest machine learning model effectively distinguished intraCCA tissue from CCA bile, identifying Rhodococcus and Ralstonia as diagnostically significant. The model also excelled in differentiating CCA bile from BBD bile, achieving an AUC value of 0.931 in external validation. Using unsupervised hierarchical clustering, we established Biletypes based on microbial signatures in our cohort. A combination of 17 genera effectively stratified patients into Biletype A and Biletype B. Biletype B robustly discerned CCA from BBD, with Sub-Biletype B1 correlating with advanced TNM stage and poorer prognosis. Among the 17 genera, bacterial Cluster 1, composed of Sphingomonas, Staphylococcus, Massilia, Paenibacillus, Porphyrobacter, Lawsonella, and Aerococcus, was enriched in Biletype B1 and predicted CCA with an AUC of 0.96. Staphylococcus emerged as a promising single-genus predictor for CCA diagnosis and staging. In conclusion, this study delineates a potential microbiota transition pathway from the gut through CCA bile to intra-CCA tissue, proposing Biletypes and Staphylococcus as biomarkers for CCA prognosis.

Objective To determine the relationship between local microenvironment and cervical cancer (CC). Discovering differential florae and metabolites associating with CC. Design Observational study. Participants 10 LSIL patients, 10 HSIL patients, 10 CC patients and 10 healthy controls were enrolled in our study. Methods We performed 16S rDNA sequencing and metabolomic analysis in the cervicovaginal fluid to reveal the differential florae and metabolites during cervical carcinogenesis. Results Carcinogenesis was associated with alterations in microbiome diversity, individual taxa, and functions. Bacterial diversity was increased and the composition was changed by the influence of cervical cancer carcinogensis. Lactobacillus was significantly reduced in LSIL and CC patients. On the contrary, the relative abundance of Prevotella was significantly elevated in LSIL patients. Lipids were significantly declined in LSIL and HSIL patients compared to HC participants, and significantly elevated in CC compared to LSIL and HSIL patients. Pathway enrichment analysis found phenylalanine, tyrosine and tryptophan biosynthesis pathway was crucial in cervical carcinogenesis. Conclusions These results showed that microbic and metabolomic profiling are capable of distinguishing CC from precancer and highlighted potential biomarkers for the early detection of cervical dysplasia. These differential microorganisms and metabolites expected to become a potential tool to assist in the diagnosis of cervical cancer. Funding We would like to thank Development & Demonstration Program of Wuxi (N20192004), Key Research & Development Program of Jiangsu Province (BE2015617), The 5th Phase of “Project 333” of Jiangsu (BRA2019024) and Innovation and Entrepreneurship Training Program for College Students in Jiangsu Province (KYCX19_1182) for their support.

Gambogic acid (GA) is a naturally active compound extracted from the Garcinia hanburyi with various anticancer activities. However, whether GA induces pyroptosis (a newly discovered inflammation-mediated programmed cell death mechanism) in ovarian cancer (OC) has not yet been reported. This study revealed that GA treatment reduced cell viability by inducing pyroptosis in OC cell lines. Typical pyroptosis morphological manifestations such as cell swelling with large bubbles and loss of cell membrane integrity, were observed. Cleaved caspase-3 and GSDME-N levels increased after GA treatment, and knocking out GSDME or using a caspase-3 inhibitor could switch GA-induced cell death from pyroptosis to apoptosis, indicating GA induced caspase-3/GSDME-dependent pyroptosis. Furthermore, this research indicated that GA significantly increased reactive oxygen species (ROS) and p53 phosphorylation. OC cells pretreated with ROS inhibitor N-Acetylcysteine (NAC) and the specific p53 inhibitor pifithrin-μ could completely reverse the pyroptosis post-treatment. Elevated p53 and phosphorylated p53 reduced mitochondrial membrane potential (MMP) and Bcl-2, increase the expression of Bax, and damage mitochondria by releasing cytochrome c to activate the downstream pyroptosis pathway. Different doses of GA inhibited tumor growth in ID8 tumor-bearing mice, and high-dose GA increased in tumor-infiltrating lymphocytes CD3, CD4, and CD8 were detected in tumor tissues. Notably, the expressions of GSDME-N, cleaved caspase-3 and other proteins were increased in tumor tissues with high-dose GA groups. These findings demonstrate that GA-treated OC cells could induce GSDME-mediated pyroptosis through the ROS/p53/mitochondria signaling pathway and caspase-3/-9 activation. Thus, GA is a promising therapeutic agent for OC treatment

Background: Immune checkpoint blockade (ICB) is considered as a promising approach for cancer treatment. However, the potency of ICB therapy in yolk sac tumors (YSTs) has not been confirmed, and the comprehensive analysis of tumor immune microenvironment and the expression of PD-1/PD-L1 and CTLA4 were also not thoroughly evaluated. Methods: Immunohistochemistry was performed in formalin-fixed, paraffin-embedded tumor specimens from 23 YSTs patients to detect the density and distribution of tumor-infiltrating T cells, tertiary lymphoid structures (TLSs), as well as the expression of PD-1/PD-L1 and CTLA4. Results: Overall, more than half (61 %) of all patients exhibited an immune-desert phenotype based on CD3+ T cells. PD-1 expression was identified in five tumor samples (21.7 %), and PD-L1 expression exhibited a different positive rate in tumor cells (TCs) and tumor-infiltrating lymphocytes (TILs) (39.1 % and 17.4 %). Noteworthily, the rate of positive CTLA4 expression in both TCs and TILs was markedly higher (69.6 % and 56.5 %) than those of PD-1 and PD-L1 expression. Furthermore, TLSs were observed in 21.74 % of all tissues, and samples with TLSs exhibited significantly higher densities of TILs and higher expression of immune checkpoint molecules, particularly PD-1/PD-L1. In addition, tumors located in testes also exhibited a higher density of TILs and higher expression of immune checkpoint molecules. Conclusion: Generally a high frequency of CTLA4 expression was found, PD-1/PD-L1 expression, the immuneinflamed phenotype, and TLSs were low frequency in YSTs, however, YSTs in testes showed a higher density of TILs and higher expression of immune checkpoint molecules.

Cervical cancer (CC) continues to be one of the most common cancers among females worldwide. It takes a few years or even decades for CC to arise in a minority of women with cervical precancers. An increasing corpus of studies today indicates that local microecology and carcinogenesis are intimately related. To investigate the changes in cericovaginal microecology with the development of cervical cancer, we performed 16S rDNA sequencing and metabolomic analysis in cericovaginal fluid from 10 LSIL patients, 10 HSIL patients, 10 CC patients and 10 healthy controls to reveal the differential flora and metabolites during cervical carcinogenesis. Carcinogenesis is associated with alterations in microbiome diversity, individual taxa, and functions with notable changes in Lactobacillus, Prevotella and Aquabacterium, as well as in cervicovaginal metabolites that correlate with cervicovaginal microbial patterns. Increased bacterial diversity and a decline in the relative abundance of Lactobacillus, the dominant species in the cericovaginal flora, are observed when cervical lesions advance. According to KEGG pathway enrichment analysis, lipids and organic acids change as cervical cancer progresses, and the phenylalanine, tyrosine, and tryptophan biosynthesis pathway is essential for the development of cervical cancer. Our results reveal that microbic and metabolomic profiling is capable of distinguishing CC from precancer and highlights potential biomarkers for the early detection of cervical dysplasia. These differential microorganisms and metabolites are expected to become a potential tool to assist in the diagnosis of cervical cancer.

目的 探讨氨肽酶2(aminopeptidase P2,XPNPEP2)在乳腺癌组织中的表达情况及其与区域淋巴结转移和分子标志物表达等病理参数的关系及其临床意义.方法 运用组织芯片技术和免疫组织化学方法检测美国Biomax公司提供的组织芯片中的69例乳腺癌组织和3例癌旁正常乳腺组织以及2013年3月至6月就诊于华中科技大学同济医学院附属同济医院的16例乳腺癌患者的癌旁正常乳腺组织中的XPNPEP2蛋白表达情况.结合临床资料分析其表达情况与肿瘤TNM分期、分级、区域淋巴结转移情况、雌激素受体(estrogen receptor,ER)、孕激素受体(progesterone receptor,PR)和人表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)的关系.通过Kaplan-Meier Plotter数据库分析不同临床病理参数下XPNPEP2表达和患者无复发生存(relapse-free survival,RFS)的关系.结果 乳腺癌组织中XPNPEP2蛋白表达水平升高,且XPNPEP2表达在肿瘤TNM分期、分级和区域淋巴结转移方面比较,差异均具有统计学意义(均P<0.05).Kaplan-Meier Plotter数据库分析显示,在淋巴结阳性、三阴性和PR阴性的乳腺癌患者中,XPNPEP2 mRNA高表达患者的10年RFS率均较低(均P<0.05).结论 乳腺癌组织中XPNPEP2蛋白表达水平升高,其高表达与区域淋巴结状态及肿瘤分子标志物表达情况密切相关,并预示预后不良.

In China, premature rupture of membranes (PROM) counts as a major pregnancy complication in China and usually results into adverse pregnancy outcomes. We analysed the vagina microbiome composition using 16S rDNA V3–V4 amplicon sequencing technology, in this prospective study of 441 women in their third trimester of pregnancy. We first divided all subjects into PROM and HC (healthy control) groups, in order to investigate the correlation of vagina microbiome composition and the development of PROM. We found that seven pathogens were higher in the PROM group as compared to the HC group with statistical significance. We also split all subjects into three groups based on Lactobacillus abundance-dominant (Lactobacillus > 90%), intermediate (Lactobacillus 30–90%) and depleted (Lactobacillus < 30%) groups, and explored nine pathogenic genera that were higher in the depleted group than the intermediate and dominant groups having statistical significance. Finally, using integrated analysis and logistics regression modelling, we discovered that Lactobacillus (coeff = −0.09, p = 0.04) was linked to the decreased risk of PROM, while Gardnerella (coeff = 0.04, p = 0.02), Prevotella (coeff = 0.11, p = 0.02), Megasphaera (coeff = 0.04, p = 0.01), Ureaplasma (coeff = 0.004, p = 0.01) and Dialister (coeff = 0.001, p = 0.04) were associated with the increased risk of PROM. Further study on how these pathogens interact with vaginal microbiota and the host would result in a better understanding of PROM development.

LIM homeobox 6 (LHX6) has been reported to be downregulated and inhibits cell proliferation in various cancers. Alternative splicing of LHX6 leads to six annotated isoforms, which can be found in the NCBI database. However, the expression patterns and potential roles of these isoforms remain poorly characterized in cervical cancer. Here, we demonstrated that the LHX6 isoforms containing exon 12 (LHX6EX(+12) group) and isoforms lacking exon 12 (LHX6EX(–12) group) were differentially expressed in cervical tissue by qRT‐PCR. The mRNA expression level of LHX6EX(+12) group was higher than that of LHX6EX(−12) group in cervical cancer tissue. Knockdown of LHX6EX(+12) group and all LHX6 isoforms (LHX6All group) inhibited cell growth, increased cell apoptosis, and induced cell cycle arrest from G0/G1 phase to S phase in vitro. Consistently, overexpression of the LHX6EX(+12) group promoted cervical cancer cell proliferation in vitro. In contrast, no significant differences in cell proliferation were found between LHX6EX(−12) isoform knockdown group and its control. RNA‐sequencing suggested that the LHX6EX(+12) isoform group might exert its cancer‐promoting effects in cervical cancer via regulating MAPK signaling pathway. Downregulation of the LHX6EX(+12) group significantly suppressed the phosphorylation of MRK, ERK, JNK, and P38 at the protein level. We also identified some unique biological processes and signaling pathways in which each isoform group might be involved. In summary, our results indicated that LHX6EX(+12) isoform group was the dominant oncogenic type of LHX6 in cervical cancer, which may be a new biomarker and a potential precise therapeutic target for cervical cancer in the future.

Cervical cancer(CC)continues to be the second leading cause of cancer death in women aged 20 to 39[1].Recent research has shown that the cervical microbiota differs in different stages of cervical carcinogenesis[2].It has been reported that the gut microbiota is linked to host metabolism,which influences the progression of the disease[3].The role of cervicovaginal flora in affecting host metabolism,thereby causing the progression of cervical cancer,requires further research.In this direction,metabolomics has been implemented to detect small molecular differential metabolites in cancer progression.Metabolomics is the study of the thousands of low-molecular-weight molecules found in biological fluids and tissues of different individuals,whether normal or afflicted with disease,and it reflects reasonable changes in biological functions.

TMTP1 is a polypeptide independently screened in our laboratory, which can target tumors in situ and metastases. In previous work, we have successfully developed a near-infrared (NIR) probe TMTP1-PEG4-ICG for tumor imaging. However, the limited ability to target tumor micrometastases hinders its further clinical application. Multimerization of peptides has been extensively demonstrated as an effective strategy to increase receptor binding affinity due to "multivalent effect" or "apparent cooperative affinity". In this study, a novel TMTP1 homodimer-directed NIR probe (TMTP1-PEG4)2-ICG was successfully constructed and synthesized. The cyclic TMTP1 peptides were bridged by two PEG4 linkers and then labeled with ICG-NHS for tumor imaging and photothermal therapy. In vivo biodistribution were assessed in normal BALB/c mice, and tumor targeting abilities of (TMTP1-PEG4)2-ICG and its monomer were evaluated and compared in 4T1-bearing subcutaneous tumor and lymph node metastasis model mice. Biodistribution analysis in vivo revealed that (TMTP1-PEG4)2-ICG was cleared mainly in both liver and kidney dependent way. Comparing with free ICG dye or TMTP1-PEG4-ICG probe, this improved (TMTP1-PEG4)2-ICG dimer showed more sensitive tumor imaging and could clearly identify tumors at a minimum volume of 10 mm3. Additionally, when compared to its monomer, lymph node (LN) metastases could also be apparently visualized and easily distinguished from normal LN by the novel dimer at 24 h post-injection. The blocking study revealed that the tumor accumulation of this probe was specifically medicated by receptor-ligand interaction. Furthermore, with the increase in stability and tumor targeting ability of ICG in vivo, the probe could also be an attractive photothermal agent to significantly inhibit tumor growth under 808 nm NIR laser irradiation. In conclusion, our work revealed that the novel (TMTP1-PEG4)2-ICG dimer could be a promising theranostic agent for sensitive tumor imaging and imaging-guided photothermal therapy, indicating its broad prospects for further clinical transformation.

Pyroptosis is a recently identified form of programmed cell death (PCD) that exerts a vital influence on the antitumor immune response. GA, a xanthone structure isolated from gamboge resin, is a naturally occurring bioactive ingredient with several anticancer activities, such as activities that affect cell cycle arrest, apoptosis, and autophagy. Here, we found that GA decreased the viability of the CRC cell lines, HCT116 and CT26, in a dose- and time-dependent manner, and multiple pores and large bubbles in the membranes, which are morphological characteristics of pyroptosis, were observed by light microscopy and transmission electron microscopy (TEM). Furthermore, the cleavage of gasdermin E (GSDME) was observed after exposure to GA, along with concomitant activation of caspase-3. Additionally, GSDME-dependent pyroptosis triggered by GA could be attenuated by siRNA-mediated knockdown of GSDME and treatment with the caspase-3 inhibitor. Moreover, we found that GA induced pyroptosis and significantly inhibited tumor growth in CT26 tumorbearing mice. Strikingly, significantly increased proportions of CD3+ T cells, cytotoxic T lymphocytes (CTLs), and dendritic cells (DCs) were observed in the tumor microenvironment in the GA-treated groups. Moreover, significantly increased proportions of CTLs and effector memory T cells (TEM) (CD8(+) CD44(+) CD62L(-)) were also detected in the spleens of the GA-treated groups, suggesting that the pyroptosis-induced immune response generated a robust memory response that mediated protective immunity. In this study, we revealed a previously unrecognized mechanism by which GA induces GSDME-dependent pyroptosis and enhances the anticancer immune response. Based on this mechanism, GA is a promising antitumor drug for CRC treatment that induces caspase-3-GSDMEdependent pyroptosis. This study provides novel insight into cancer chemoimmunotherapy.

细胞焦亡是由含半胱氨酸的天冬氨酸蛋白水解酶(caspase)家族蛋白介导的炎症性程序化细胞死亡,其特点是细胞膜上形成质膜孔,导致细胞膜破裂,释放炎性介质.细胞焦亡途径分为经典途径以及非经典途径,其与多种恶性肿瘤的发生发展密切相关,现阶段利用细胞焦亡进行抗肿瘤治疗也取得了极大进展.本文就细胞焦亡的发现、途径、细胞焦亡与妇科恶性肿瘤发生发展的关系及其对妇科恶性肿瘤治疗作用的研究进展做一简要综述.

BACKGROUND:Premature rupture of membranes (PROM) is a major pregnancy complication in China and usually leads to adverse pregnancy outcomes. The major aim of this study was to search for microorganisms and their related metabolites that have direct relationship with PROM.METHODS:For vaginal discharge samples, metagenomics sequencing was applied to identify microorganisms that were enriched in PROM subjects, and untargeted metabolomics was applied to characterize the metabolites changes in PROM subjects compared to healthy controls (HC). Correlation analysis was then used to explore the relationship between these microorganisms and metabolites changes.RESULTS:Two upstream metabolites of glycolysis, N-acetyl-D-galactosamine (GalNAc) and sucrose, were found downregulated in the PROM group (P=0.04 and P=0.041, respectively). Higher percentages of conditional pathogens, such as of Streptococcus (8.4% vs. 6.1% in HC group, P=0.15) and Chlamydia (4.3% vs. 2.3% in HC group, P=0.07) were found in PROM group. Other common conditional pathogens including Prevotella, Staphylococcus, Mycobacterium and Enterobacter, were also higher in PROM group, although their absolute percentages were low and the differences did not reach statistical significance due to relative small sample size. Correlation analysis further demonstrated a positive correlation of downregulation of glycolysis metabolites with higher percentage of conditional pathogens.CONCLUSIONS:Integrated metagenomics and metabolomics analysis can be used to track the subtle changes in the vaginal microenvironment. Downregulation of glycolysis substrates (GalNAc and sucrose) and increase of related pathogenic microorganisms (Streptococcus and Chlamydia) could serve as early warning biomarkers of PROM.