Background:Precision oncology in non-small cell lung cancer (NSCLC) requires comprehensive genomic characterization to inform therapeutic decisions. However, the spectrum of genomic alterations and their co-mutation patterns, particularly in relation to immunotherapy biomarkers, remains incompletely understood. This study investigated the associations between driver gene alterations and clinicopathological characteristics, and characterized co-mutation patterns in relation to programmed cell death ligand 1 (PD-L1) expression and tumor mutational burden (TMB) levels. Methods:We conducted a retrospective analysis of 431 NSCLC patients, utilizing a comprehensive pan-solid tumor next-generation sequencing (NGS) panel covering 654 genes to characterize genomic alterations. Clinicopathological characteristics were systematically collected and analyzed to identify significant differences across various mutational profiles. Systematic assessments were performed to evaluate interactions between co-mutations and PD-L1 expression as well as TMB. Results:Genomic profiling revealed EGFR as the most frequently mutated driver gene (59.1%), followed by TP53 (39.7%), RBM10 (14.6%), MUC16 (12.7%), and KRAS (10.0%). EGFR mutations showed a strong female predominance (P<0.001), whereas MUC16 (P=0.004) and KRAS (P<0.001) alterations were more common in males. TP53 (P=0.005) and KRAS (P<0.001) alterations were more frequently found in smokers. Patients with advanced disease stages exhibited higher frequencies of TP53 mutations (P<0.001), contrasting with the predominance of EGFR (P<0.001) and RBM10 (P=0.04) mutations in early-stage tumors. TP53 co-mutations were the most common types, especially TP53-EGFR and TP53-KRAS co-mutations. TP53-EGFR co-mutations had higher PD-L1 expression (P=0.03) and TMB level (P=0.03) than EGFR-only mutations. Patients with TP53-KRAS co-mutations showed significantly higher PD-L1 expression (P=0.01) and TMB level (P=0.01) than KRAS-only mutations. Conclusions:This study underscores the diverse genetic mutations occurring in NSCLC patients with varying risk factors. The identification of TP53 co-mutations provides critical insights for personalized immunotherapeutic strategies and optimizing treatment outcomes.

BACKGROUND:Uric acid, the final product of purine metabolism, is primarily excreted through the kidneys and intestines. Dysregulation in uric acid production or excretion can result in hyperuricemia and gout. Several proteins play important roles in uric acid metabolism. Among them, GLUT9, a key protein for uric acid excretion, has garnered significant attention, particularly for two SNPs (rs3733591 and rs1014290) on its encoding gene SLC2A9. However, their relationship with gout and hyperuricemia in the Han Chinese population has not been researched. METHODS:This study investigated 498 individuals, including 300 patients with hyperuricemia or gout and 198 healthy controls. Serum uric acid levels were measured, and the genotypes of rs3733591 and rs1014290 were determined using the multicolor melting curve analysis (MMCA) method. These results were subjected to statistical analysis. RESULTS:This detection result highlights the value of MMCA as a rapid and accurate method for genotyping rs3733591 and rs1014290. Statistical analysis revealed that the proportion of the wild-type (C) allele at rs3733591 and the mutant (A) allele at rs1014290 were significantly higher in patients with hyperuricemia and gout compared to healthy controls. Additionally, individuals with the homozygous mutant genotype at rs3733591 had significantly lower uric acid levels compared to those with the wild-type and heterozygous genotypes, whereas homozygous mutants of rs1014290 exhibited higher uric acid levels. Among the various models based on clinical and laboratory data from 433 participants, the logistic regression model based on eight factors - gender, age, eGFR, WBC, HDL, MCHC, rs3733591, and rs1014290 - demonstrated the best diagnostic performance for gout and hyperuricemia, achieving an AUC of 0.8737. CONCLUSION:In conclusion, this study demonstrated a close association between SLC2A9 gene polymorphisms and gout as well as hyperuricemia in the Han Chinese population. Furthermore, the SLC2A9 gene polymorphisms could serve as key parameters for diagnosing these conditions.

INTRODUCTION BRCA1/2 germline mutations are the most well-known genetic determinants for breast cancer. However, the distribution of germline mutations in non-BRCA1/2 cancer susceptibility genes in Chinese breast cancer patients is unclear. The association between clinical characteristics and germline mutations remains to be explored. METHODS Consecutive breast cancer patients from Peking University People's Hospital were enrolled. Clinical characteristics were collected and next-generation sequencing (NGS) was performed using blood samples of participants to identify pathogenic/likely pathogenic (P/LP) germline mutations in 32 cancer susceptibility genes including homologous recombination repair (HRR) genes. RESULTS A total of 885 breast cancer patients underwent the detection of germline mutations. 107 P/LP germline mutations of 17 genes were identified in 116 breast cancer patients including 79 (8.9%) in BRCA1/2 and 40 (4.5%) in 15 non-BRCA1/2 genes. PALB2 was the most frequently mutated gene other than BRCA1/2 but still relatively rare (1.1%). There were 38 novel P/LP germline variants detected. P/LP germline mutations in BRCA1/2 were significantly associated with onset age(P<0.001), the family history of breast/ovarian cancer(P=0.010), and molecular subtype(P<0.001), while correlated with onset age (P<0.001), site of breast tumor (P=0.028) and molecular subtype (P< 0.001) in HRR genes. CONCLUSIONS The multiple-gene panel prominently increased the detection rate of P/LP germline mutations in 32 cancer susceptibility genes compared to BRCA1/2 alone. Onset younger than or equal to 45 years of age, bilateral and triple-negative breast cancer (TNBC) patients may be more likely to be recommended for detecting P/LP germline mutations in HRR genes.

The SLC26A4 gene is the second most common cause of hereditary hearing loss in human. The aim of this study was to utilize the minigene assay in order to identify pathogenic variants of SLC26A4 associated with enlarged vestibular aqueduct (EVA) and hearing loss (HL) in two patients. The patients were subjected to multiplex PCR amplification and next-generation sequencing of common deafness genes (including GJB2, SLC26A4, and MT-RNR1), then bioinformatics analysis was performed on the sequencing data to identify candidate pathogenic variants. Minigene experiments were conducted to determine the potential impact of the variants on splicing. Genetic testing revealed that the first patient carried compound heterozygous variants c.[1149 + 1G > A]; [919–2 A > G] in the SLC26A4 gene, while the second patient carried compound heterozygous variants c.[2089 + 3 A > T]; [919–2 A > G] in the same gene. Minigene experiments demonstrated that both c.1149 + 1G > A and c.2089 + 3 A > T affected mRNA splicing. According to the ACMG guidelines and the recommendations of the ClinGen Hearing Loss Expert Panel for ACMG variant interpretation, these variants were classified as “likely pathogenic”. This study identified the molecular etiology of hearing loss in two patients with EVA and elucidated the impact of rare variants on splicing, thus contributing to the mutational spectrum of pathogenic variants in the SLC26A4 gene.

Objective To explore the role of G protein pathway suppressor 2(GPS2)in regulating proliferation, migration and invasion in hepatocellular carcinoma cell line MHCC-97H.Methods The expression of GPS2 in hepatocellular carcinoma (HCC) cells were detected using quantitative PCR or Western blot; GPS2 was either over-expressed or knocked down in HCC cells using over-expression vectors or short hairpin RNAs (shRNAs) for GPS2, respectively. Clone formation assay, scratch assay and Transwell assay were used to detect the effects of GPS2 on proliferation, migration and invasion of HCC cells. Results In HCC cells, over-expression of GPS2 was found to inhibit the proliferation, migration and invasion of HCC cells MHCC-97H, while knockdown of GPS2 up-regulated proliferation, migration and invasion of MHCC-97H(P＜0.05). The over-expression of GPS2 in MHCC-97H cells was found to inhibit the expression of cell proliferation, migration and invasion-related factors such as EGFR, MMP1, MMP3 and MMP9, while knockdown of GPS2 upregulated the expression of these factors(P＜0.05). Conclusions GPS2 is a novel HCC regulatory factor that inhibits proliferation, migration and invasion of HCC cells.

The early diagnosis of lung cancer has become the focus of clinical attention, with the incidence and mortality of lung cancer increasing. Aberrant DNA methylation occurs in the primary stage of lung cancer, then the methylation degree can be changed dynamically due to the progress and the treatment of lung cancer. To date, a growing number of studies have reported that special gene methylation exploits in the clinical diagnosis, curative effect monitoring, and prognosis evaluation of lung cancer. Meanwhile, clinical trials about DNA methyltransferase inhibitors for lung cancer therapy are also underway. It is worth looking forward that detecting aberrant DNA methylation helps diagnose and treat lung cancer.

UFMylation is a ubiquitination-like modification that is related to endoplasmic reticulum stress and unfolded protein response. A recent study reported that Ufl1, a key enzyme of UFMylation, protects against heart failure, indicating that UFMylation may be associated with heart function regulation. In the present study, we initially constructed a Flag-6×His-tagged Ufm1ΔSC transgenic (Tg-Ufm1) mouse model that enables UFMylation studies in vivo. Tg-Ufm1 mice showed significant activation of UFMylation in hearts. By using this model, we identified 38 potential Ufm1-binding proteins in heart tissues through LC‒MS/MS methods. We found that these proteins were associated with mitochondria, metabolism and chaperone binding. By using transcriptomic screening, we identified Tnfaip2 as a novel UFMylation-associated gene. Overexpression of Ufm1 significantly upregulated the protein expression of Tnfaip2, whereas isoproterenol treatment decreased Tnfaip2 expression in Tg-Ufm1 mice. These data may provide novel clues for UFMylation in cardiac hypertrophy.

Breast cancer (BC) is associated with hereditary components, and some deleterious germline variants have been regarded as effective therapeutic targets. We conducted a clinic-based, observational study to better understand the distribution of deleterious germline variants and assess any clinicopathological predictors related to the variants among Chinese BC patients using a 32 cancer-related genes next-generation sequencing panel. Between November 2020 and February 2022, a total of 700 BC patients were recruited, and 13.1% (92/700) of them carried deleterious germline variants in 15 cancer-related genes, including 37 (37/700, 5.3%) in BRCA2, 29 (29/700, 4.1%) in BRCA1, 8 (8/700, 1.1%) in PALB2, 4 (4/700, 0.6%) in NBN, 3 (3/700, 0.4%) in MRE11A, 3 (3/700, 0.4%) in TP53 and 12 (12/700, 1.7%) in other genes. There were 28 novel variants detected: 5 in BRCA1, 14 in BRCA2, and 9 in non-BRCA1/2 genes. The variants in panel genes, HRR (homologous recombination repair)-related genes, and BRCA1/2 were significantly associated with the following clinicopathological factors: age at the initial diagnosis of BC, family history of any cancer, molecular subtype, Ki-67 index, and hereditary risk. In conclusion, we further expanded the spectrum of germline deleterious variants in Chinese BC patients, and the clinicopathological predictors of variants were identified to facilitate clinical genetic testing and counseling for appropriate individuals.

Objective: To investigate the germline mutation status of related genes in breast cancer patients and high-risk individuals by next-generation sequencing. To analyze the correlations between homologous recombination repair (HR) pathway gene mutation status and clinicopathological characteristics of breast cancer patients. To supplement the database of breast cancer related gene mutations in Chinese population. Methods: This study is a cross-sectional study. From October 2020 to September 2021, whole blood samples were collected from 350 breast cancer patients and 49 high-risk individuals, admitted to Peking University People's Hospital and accepted genetic testing voluntarily. Germline mutations in 32 breast cancer related genes were detected by NGS. The clinicopathological characteristics, including age at the onset, family history, unilateral/bilateral tumor, Luminal typing (Luminal A subtype, Luminal B subtype, HER2-enriched subtype and triple negative breast cancer), tumor size and metastasis, were analyzed, and the correlations between HR pathway gene mutation status and clinicopathological characteristics were analyzed by Chi-squared test and Fisher's exact probability test. Results: Among 350 breast cancer patients, 64 (18.3%) cases carried gene pathogenic mutations (including pathogenic and likely pathogenic mutations), including 47 (13.4%) in BRCA1/2, 16 (4.6%) in non-BRCA1/2 genes, 1 (0.3%) in BRCA2 and FANCL. Among 49 high-risk individuals, 7 (14.3%) cases carried gene pathogenic mutations, including 6 (12.3%) in BRCA1/2 and 1 (2%) in ATM genes. BRCA1/2 pathogenic mutations were associated with age at the onset (18%, 8.7%, χ²=6.346, P=0.012), and the BRCA1/2 pathogenic mutation frequency was higher in patients diagnosed at age ≤45 years. HR pathway gene mutations (including pathogenic, likely pathogenic and uncertain significance mutations) were correlated with unilateral/bilateral tumor (49.5%, 68.4%, χ²=4.841, P=0.028) and Luminal typing (45.7%, 62.2%, 32%, 60%, χ²=12.004, P=0.007), and the HR mutation frequencies were higher in patients with bilateral tumor, Luminal B breast cancer and triple negative breast cancer (TNBC). Conclusion: The BRCA1/2 pathogenic mutation frequency in high-risk individuals is similar to that in breast cancer patients, and BRCA1/2 testing is helpful to guide breast cancer screening and prevention in high-risk individuals. Patients with early onset breast cancer, bilateral breast cancer, Luminal B breast cancer and TNBC have higher mutation frequencies of HR pathway genes, and HR pathway genes testing should be conducted as soon as possible to provide laboratory evidence for diagnosis, treatment, prognosis and risk evaluation of breast cancer.

目的 探讨UFM化修饰在心肌肥厚小鼠中的变化,并筛选潜在泛素折叠修饰因子1(UFM1)结合蛋白.方法 取8周龄C57BL/6J小鼠,背部皮下注射5 mg/(kg·d)的异丙肾上腺素(ISO)构建心肌肥厚模型.采用免疫荧光染色检测UFM1的表达水平及定位情况.采用蛋白免疫印迹法检测肥厚心肌组织中UFM化修饰变化及相关酶UFC1和UFL1的表达.通过免疫沉淀联合蛋白质谱筛选肥厚心肌组织内UFM1的结合蛋白.结果 在ISO诱导的心肌肥厚小鼠模型中,UFM1的表达水平显著增加.在肥厚心肌组织内,UFM化修饰水平升高,且伴有UFC1和UFL1表达水平的下调,提示心肌肥厚时UFM化修饰调控紊乱.采用蛋白质组学分析,共筛选到21种潜在的UFM1结合蛋白,其中FABP5、TNNT2、NCAM1、S100A8等可能介导了 UFM化修饰与心肌肥厚之间的关联.结论 UFM化修饰在心肌肥厚小鼠中被激活,可能参与心肌肥厚发病的调控.

Background and aims: Triple-negative breast cancer (TNBC) is a subtype of breast cancer with a poor prognosis due to its aggressive biological behavior and lack of therapeutic targets. Here, we aimed to identify specific biomarkers for TNBC by using RNA-sequencing and bioinformatics analysis.Materials and methods: Fresh breast tumor tissues were obtained from 34 patients who were admitted to the Breast Center, Peking University People's Hospital, from June 2020 to December 2020; the patients were pathologically diagnosed with primary breast cancer and underwent surgery for the resection of tumor tissues. Tumor-tissue RNA was extracted and the generated cDNA libraries were sequenced using the NextSeq platform, after which the differentially expressed genes (DEGs) between TNBC and other subtypes of breast cancer were identified and DEG functional-enrichment analysis was performed. Next, weighted gene co-expression network analysis (WGCNA) was used to identify the most significant module and hub genes in TNBC, and then the correlations between the hub genes and the prognosis of TNBC patients were analyzed through survival analysis. Lastly, qRT-PCR analysis was used to validate the expression levels of hub genes in tumor tissues from TNBC and other subtypes of breast cancer.Results: Comparison of TNBC tissues and tissues from other subtypes of breast cancer led to the identification of 273 DEGs in TNBC: 172 upregulated and 101 downregulated genes. In Gene Ontology analysis of the DEGs, five terms were significantly enriched, "developmental process," "anatomical structure development," "tissue development," "cell cycle," and "epithelium development," and in Kyoto Encyclopedia of Genes and Genomes pathway analysis, the most significantly enriched pathways for all DEGs were "cell cycle," "mitophagy-animal," and "autophagy-animal." Furthermore, we identified the core module related to TNBC and screened for hub genes by using WGCNA, and after verifying the top 100 genes based on survival analysis, we selected four genes as the hub genes: SERPINB4, SMR3A, FERMT1, and STARD4; elevated expression of these genes was associated with poor overall survival (OS) of TNBC patients. Notably, qRT-PCR results indicated that FERMT1 mRNA expression was significantly upregulated in TNBC samples. Conclusion: The DEG profiles between tissues from TNBC and other subtypes of breast cancer were identified using RNA-sequencing and bioinformatics analysis. FERMT1 was significantly upregulated in TNBC tumor tis-sues, and increased expression of FERMT1 was associated with poor OS of TNBC patients. FERMT1 could serve as a specific biomarker of and therapeutic target in TNBC.

BACKGROUND Long noncoding RNA (lncRNA) ZNFX1-AS1 (ZFAS1) is a newly discovered lncRNA, but its diagnostic value in gastric cancer is unclear. AIM To investigate the potential role of ZFAS1 in gastric cancer and to evaluate the clinical significance of ZFAS1 as a biomarker for gastric cancer screening. METHODS Quantitative real-time polymerase chain reaction (qRT-PCR) was used to screen for gastric cancer-associated lncRNAs in gastric cancer patients, gastric stromal tumor patients, gastritis or gastric ulcer patients, and healthy controls. Correlations between ZFAS1 expression and clinicopathological features were analyzed. The biological effects of ZFAS1 on the proliferation, migration, and invasion of gastric cancer cells were studied by MTT, colony formation, and transwell mi-gration assays. The potential mechanism of ZFAS1 was demonstrated using enzyme-linked immunosorbent assay and qRT-PCR. The relationship between ZFAS1 and tumorigenesis was demonstrated using in vivo tumor formation assays. RESULTS The plasma level of lncRNA ZFAS1 was significantly higher in preoperative patients with gastric cancer than in individuals in the other 4 groups. Increased expression of ZFAS1 was significantly associated with lymph node metastasis, advanced TNM stage, and poor prognosis. ZFAS1 regulated the proliferation, migration, and invasion of gastric cancer cells and regulated the growth of gastric cancer cells in vivo. LIN28 and CAPRIN1 were identified as key downstream mediators of ZFAS1 in gastric cancer cells. CONCLUSION LncRNA ZFAS1 promoted the invasion and proliferation of gastric cancer cells by modulating LIN28 and CAPRIN1 expression, suggesting that ZFAS1 can be used as a potential diagnostic and prognostic biomarker in gastric cancer.

OBJECTIVE:Breast cancer is recognized as the most common cause of malignancy and cancer death worldwide; however, mutations in the cancer-related BRCA genes are detected in only 2-3% of patients with breast cancer. Because next-generation sequencing technology allows concurrent sequencing of numerous target genes, diverse cancer-susceptibility genes are now being evaluated, although their significance in clinical practice remains unclear.METHODS:In this study, we developed a sequencing panel containing the genes BRCA1, BRCA2, TP53, PIK3CA, ERBB2 (Her2), and PTEN, which are all associated with cancer risk in patients, and we enrolled 60 patients with breast cancer.RESULTS:Germline mutations were found to be carried by nine patients (15%): 3 in BRCA1, 5 in BRCA2, and 1 in TP53. The patients harboring these mutations are considered to face a high risk of developing malignant tumors, and cancer screening is thus recommended for the patients.CONCLUSION:This study demonstrates the feasibility of using Ion Torrent sequencing technology for reliably detecting gene mutations in clinical practice for guiding individualized drug therapy or combination therapies for breast cancer.

Objective:To investigate the germline mutation status in multi-pathway in Chinese female breast cancer patients and explore their correlation with clinicopathological characteristics. Aim to enrich the database of breast cancer germline gene mutations in Chinese population and provide laboratory evidence for the application of breast cancer targeted drugs.Methods:From January 2017 to July 2019, whole blood samples were collected from 148 women (age of onset concentrated in the 24~80 years old) diagnosed pathologically with breast cancer in the Department of breast surgery, Peking University People′s Hospital. Germline mutations in HR, MMR, BER, and KDR pathway related genes were detected by next-generation sequencing. The pathogenicity interpretation was performed, and pathogenic, likely pathogenic, and mutations of uncertain significance were screened. The clinicopathological characteristics including age at the onset, luminal typing, tumor size, metastasis, and family history were analyzed, and the correlation between mutations in different pathway genes and clinicopathological characteristics was analyzed by the Chi-squared test and Fisher′s exact probability test.Results:Among the 148 patients, there were 69 cases of HR mutations (including three types of mutations, including pathogenic, likely pathogenic and uncertain significance), 16 cases of MMR mutations, 6 cases of BER mutations and 8 cases of KDR mutation. ATM mutations in the HR pathway were associated with luminal typing ( P=0.054), and patients with HER2+breast cancer were more likely to carry ATM mutations. PMS2 mutations in the MMR pathway were correlated with tumor size ( P=0.060), and patients with tumor size>50 mm were more likely to carry PMS2 mutations. KDR mutations was significantly correlated with luminal typing and family history. ( P=0.021, P=0.024). Conclusion:The mutation frequency in BER, KDR, MMR and HR pathways in Chinese breast cancer patients increased successively. Germline mutations in ATM, PMS2 and KDR genes may be involved in the development of breast cancer in the Chinese population. Multi-pathway gene detection of breast cancer can provide laboratory evidence for the use of PARP inhibitors, trastuzumab and other targeted drugs.

Breast cancer is the most frequent cancer among women worldwide. Patients carrying mutations in breast cancer susceptibility genes like BRCA1 and BRCA2 (BRCA1/2) account for 5-10% of all breast cancer patients. Therefore, screening for susceptibility genes may reduce the incidence of breast cancer and improve prognosis. To provide evidence for mutation interpretation and targeted drug use in breast cancer patients, gene mutations were screened in 78 women diagnosed with sporadic breast cancer using a next-generation sequencing panel, confirmed by Sanger sequencing. Then the pathogenicity of the identified novel variants was explored using in vitro experiments including western blotting, co-immunoprecipitation and cell-migration assays. Four novel variants (BRCA2 L1390W, BRCA2 Glu432fs, BRCA1 P706L, and BRCA1 Cys882fs) were identified. BRCA2 Glu432fs decreased the expression of BRCA2 protein, enhanced cell migration and invasion ability, and prevented the protein from interacting with RAD51, resulting in a defect in the homologous recombination pathway. The identification of these novel BRCA variants and the confirmation of their pathogenicity have enriched the genetic database of breast cancer, especially in the Chinese population. Moreover, the variants are the genetic risk factors for hereditary breast cancer. Therefore, BRCA variant detection and genetic counseling for breast cancer patients are meaningful and important.

目的 研究淫羊藿苷对载脂蛋白E(apolipoprotein E,ApoE)-/-小鼠动脉粥样硬化易损斑块细胞凋亡及核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)/抗氧化反应元件(anti-oxidative response element,ARE)通路的调控作用.方法 ApoE-/-小鼠随机分为对照组、模型组和淫羊藿苷组,采用高脂喂养+右侧颈总动脉外置管术(perivascular carotid collar place-ment,PCCP)制备动脉粥样硬化易损斑块模型,给予淫羊藿苷灌胃干预.检测右侧颈总动脉病理改变,血管平滑肌细胞凋亡,Bcl-2、Bax、Caspase-3、Nrf2和ARE的表达水平及MDA、8-OHdG和TAC的水平.结果 与对照组比较,模型组出现了典型的易损斑块病理改变,细胞凋亡率,Bax、Caspase-3的表达水平及MDA和8-OHdG的水平明显升高,Bcl-2、Nrf2和ARE的表达水平及TAC的水平明显降低(P＜0.05);与模型组比较,淫羊藿苷组斑块缩小且趋于稳定,细胞凋亡率,Bax、Caspase-3的表达水平及MDA和8-OHdG的水平明显降低,Bcl-2、Nrf2和ARE的表达水平及TAC的水平明显升高(P＜0.05).结论 淫羊藿苷能够改善ApoE-/-小鼠的动脉粥样硬化易损斑块,此作用与抑制细胞凋亡及Nrf2/ARE通路有关.

Abstract Background While surgical reduction with adjuvant chemotherapy is the traditional treatment for high-grade serous ovarian cancer (HGSOC), neoadjuvant chemotherapy (NACT) has increasingly been applied. This work aims to investigate the expression profiles before and after NACT, explore changes in the tumor microenvironment, expand current treatments, and design a combination of treatment options for patients. Methods We downloaded 326 pre-NACT RNA sequencing data and 37 matched pre- and post-NACT samples from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Differentially expressed genes (DEGs) were determined with EdgeR, and Gene Ontology analysis was performed to identify the clusters responsible for the biological processes and pathways of HGSOC. Immune infiltration was analyzed using Single-sample Gene Set Enrichment Analysis (ssGSEA) and CIBERSORT. Kaplan-Meier (KM) survival analysis was performed to assess prognosis, and the potential correlations between modules and phenotypes were explored using weighted gene co-expression network analysis (WGCNA). Results After NACT, a total of 352 genes showed significant changes in RNA expression, among which 180 genes were up-regulated and 172 down-regulated. The most influential pathway was the positive regulation of mitogen-activated protein kinase (MAPK) cascade. Correlation analysis and KM survival analysis showed that overexpression of MAPK cascade genes correlated with shorter survival time in HGSOC patients. ssGSEA results showed that the expressions of anti-tumor cells (central memory CD4+ T cell and central memory CD8+ T cell) and pro-tumor cells (neutrophil and dendritic cells) were significantly increased after NACT. CIBERSORT showed that the abundances of memory B cells, NK cells, and monocytes were increased and the abundance of plasma cells was decreased after NACT. WGCNA and KM survival analysis showed that a lower abundance of Regulatory T cells (Tregs) was correlated with a better prognosis. Conclusions Gene expression of the MAPK pathway is up-regulated and the abundance of CD4+ T regulation cell decreases after NACT. Thus, the MAPK pathway may promote the differentiation of CD4+ T cells into Th17 cells while inhibiting Tregs development. The inhibited Tregs' development can lead to a better prognosis. Therefore, it is speculated that Tregs inhibitors combined with platinum-based NACT are potential treatment options for HGSOC.

Background: Mutations in the F10-coding gene can cause factor X (FX) deficiency, leading to abnormal coagulation activity and severe tendency for hemorrhage. Therefore, identifying mutations in F10 is important for diagnosing congenital FX deficiency. Methods: We studied a 63-year-old male patient with FX deficiency and 10 of his family members. Clotting and immunological methods were used to determine activated partial thromboplastin time (aPTT), prothrombin time (PT), thrombin time (TT), fibrinogen levels, FX activity, and FX antigen levels. The platelet count was determined. A mixing study was performed to eliminate the presence of coagulation factor inhibitors and lupus anticoagulant. Mutations were searched using whole-exome sequencing and certified by Sanger sequencing. Results: Genetic analysis of the proband identified two single-base substitutions: c.1085G>A (p.Ser362Asn) and c.1152C>A (p.Tyr384Ter, termination codon, caused by the DNA sequence TAA). His FX activity and antigen levels were 1.7% and 408.53 pg/mL, respectively; aPTT and PT were 52.3 and 48.0 s, respectively. One brother had the same compound heterozygous mutations, and his FX activity and antigen levels were 1.3% and 465.47 pg/mL, respectively; his aPTT and PT were 65.2 and 54.5 s, respectively. His mother, another brother, and one sister were heterozygous for c.1085G>A (p.Ser362Asn), and his daughter and grandson (6 years old) were heterozygous for c.1152C>A (p.Tyr384Ter). Conclusion: The heterozygous variants p.Ser362Asn or p.Tyr384Ter indicate mild FX deficiency, but the compound heterozygous mutation of the two causes severe congenital FX deficiency and bleeding. Genetic analysis of these two mutations may help characterize the bleeding tendency and confirm congenital FX deficiency.

Summary Inducible co-stimulator-positive (ICOS) and programmed cell death 1-positive (PD-1) are important markers for follicular helper T cells (Tfh); however, their roles and clinical values in ulcerative colitis (UC) remain unknown. In this study, we recruited 68 UC patients and 34 healthy controls. Circulating ICOS+, PD-1+ and ICOS+PD-1+ Tfh subsets were analyzed by flow cytometry. Twelve active UC patients achieving remission after treatment with 5-aminosalicylic acid were followed-up and Tfh subset changes were analyzed. Serum immunoglobulin (Ig)G, C-reactive protein (CRP), interleukin (IL)-4 and IL-21 levels and B cell subsets were analyzed and Mayo scores were calculated. Correlation analyses were performed between Tfh subsets and the clinical indicators. Receiver operating characteristic (ROC) curves were generated to evaluate the efficiency of Tfh subsets for disease monitoring. We found that levels of ICOS+, PD-1+ and ICOS+PD-1+ Tfh cells were significantly increased in active UC and significantly decreased when achieving clinical remission. Activated ICOS+PD-1+Tfh cells were positively correlated with serum CRP and Mayo scores. Furthermore, ICOS+PD-1+ Tfh cells were significantly correlated with circulating new memory B cells and plasmablasts, as well as serum IgG, IL-4 and IL-21. ROC analyses showed that when ICOS+PD-1+ Tfh cells were used in combination with PD-1+ Tfh cells, the diagnostic efficacy in distinguishing active UC from stable remission patients was higher than that of any one used alone, with area under curve (AUC) value 0·931. Our findings suggest that increased ICOS+PD-1+ Tfh cells are associated with the activation of B cells in the pathogenesis of UC, and may be a potential biomarker for UC disease monitoring.

Background: Loss of immune homeostasis to enteric pathogens is considered to be involved in the pathogenesis of ulcerative colitis (UC), and regulatory T cells (Tregs) are key for this immune homeostasis. Helios exhibits an important effect on regulating the suppressive function of Tregs. Epstein-Barr virus (EBV) is more commonly detected in UC. However, whether there is an association between EBV infection and Helios+Tregs and its impact on disease activity of UC remain unclear. We aimed to explore the clinical significance of Helios+Tregs and their potential association with EBV infection in UC. Methods: Seventy-six UC patients and 38 controls were consecutively enrolled. Helios+FoxP3+Tregs were analyzed using flow cytometry and compared among groups. Eight active UC patients treated with 5-aminosalicylic acid were followed up. Correlation analyses were conducted between Helios+FoxP3+Tregs and disease activity indicators. In addition, EBV viral loads in the mucosal lesion were quantified in active UC by quantitative polymerase chain reaction and were comprehensively analyzed in subgroups of different disease severity, and their associations with Helios+FoxP3+Tregs were also analyzed. Results: Helios+FoxP3+Tregs were significantly decreased in active UC and were inversely correlated with serum C-reactive protein and Mayo score. Moreover, we observed the recovery of Helios+FoxP3+Tregs in followed-up active UC achieving remission after treatment. EBV loads were higher in active UC, and levels of Helios+FoxP3+Tregs in the EBV-positive subgroup were lower than the EBV-negative subgroup in moderate and severe active patients. Most importantly, we found that Helios+FoxP3+Tregs were significantly negatively correlated with EBV viral loads. Conclusion: Helios+FoxP3+Tregs are likely to play a pivotal role in disease activity of UC and may be influenced by EBV infection.