Zinc finger protein 24 (ZNF24) is a conserved multifunctional transcription factor associated with tumorigenesis, but its function in bladder carcinogenesis remains unclear. Herein, the expression of ZNF24 was decreased in bladder cancer (BC) cells and tissues, and patients with higher expression of ZNF24 had a better prognosis. Doxycycline-induced overexpression and knockdown of ZNF24 identified its anti-proliferative and anti-metastasis role in BC in vitro and in vivo. The potential genes for the anti-cancer role of ZNF24, involving transcriptional regulation of several factors, such as dual-specificity phosphatase 1 and squalene epoxidase. E2 conjugating enzyme UBC9 and small ubiquitin-like modifier (SUMO) 1 were found to interact with ZNF24, suggesting that ZNF24 may be SUMOylated. Consistent with the expression, ZNF24 SUMOylation levels were decreased in BC cells and tissues. Pan-SUMOylation inhibition promoted protein degradation of ZNF24. UBC9 SUMOylated ZNF24 at Lys-27 (K27) site with SUMO1 modification and the K27 mutation of ZNF24 greatly damaged the protein stability of ZNF24. Cullin 3 (CUL3), a E3 ubiquitin ligase, was responsible for the degradation of ZNF24. ZNF24 SUMOylation prevented CUL3-mediated protein degradation of ZNF24. Overall, the crosstalk between the SUMOylation and ubiquitination of ZNF24 may be a novel regulatory mechanism to block tumorigenesis and development of BC.

The high metastasis rate is the primary contributor to the high mortality rate associated with muscle-invasive bladder cancer (MIBC). Therefore, elucidating the mechanisms involved and identifying potential therapeutic targets are crucial for improving the overall prognosis of bladder cancer (BLCA). We used consensus clustering and differential gene expression analyses to identify the key gene desmoglein 3 (DSG3). Subsequently, we examined the expression of DSG3 in BLCA and its association with the clinical characteristics and prognosis. Comprehensive in vitro and in vivo experiments were conducted to elucidate the functions of DSG3 and the reasons behind the upregulation of DSG3 in BLCA, as well as to investigate the mechanisms by which DSG3 promotes metastasis. DSG3 was markedly upregulated in BLCA, particularly in the basal/squamous (Ba/Sq) subtype. Importantly, elevated DSG3 levels demonstrated a strong association with aggressive tumor behavior and poorer clinical outcomes. Functional experiments revealed that DSG3 knockdown significantly impeded cancer stemness characteristics, epithelial-mesenchymal transition (EMT), migration, and invasion capabilities in vitro, whereas in vivo studies showed marked reductions in tumorigenesis and lung metastasis. Mechanistic investigations indicated that STAT3 transcriptionally activated DSG3 expression in BLCA cells. Downstream pathway analysis further showed that DSG3 promoted AKT phosphorylation, thereby inhibiting GSK3β activity. This molecular pathway promoted β-catenin nuclear translocation, thereby triggering transcriptional upregulation of SOX2 and MMP7 expression, ultimately mediating BLCA progression. Our study demonstrates a novel mechanism by which DSG3 enhances cancer stemness, EMT, migration, and invasive capabilities through upregulation of SOX2 and MMP7 expression through the AKT/GSK3β/β-catenin pathway, ultimately leading to growth and metastasis of BLCA. This study elucidated the role of DSG3 in BLCA and its mechanism in activating the Wnt/β-catenin signaling pathway. We anticipate this study will identify potential biomarkers for predicting progression and for assessing prognosis. Furthermore, this study introduced a novel intervention target for BLCA treatment.

Dear Editor, Bladder cancer (BLCA) is the most common malignant tumour of the urinary system and has a high recurrence rate.1, 2 N1-methyladenosine (m1A) methylation is a key mechanism of post-transcriptional regulation.3-5 m1A levels in the urine of BLCA patients are higher than in the urine of normal people, and TRMT61A is highly expressed in human BLCA tissues.6, 7 Although some reports are consistent with our previous results,8 the biological functions and potential mechanisms of action of m1A methylation in BLCA remain unknown. Here, the role of TRMT61A in BLCA was assessed in vitro and in vivo. Silencing TRMT61A inhibited 5637 cell proliferation, clonogenicity, migration and invasion, while its overexpression was promoted in T24 cells (Figure S1A,B and Figure 1A–F). Total m1A levels in RNA decreased and increased following TRMT61A knocking-down, and over-expression, respectively. Quantitative ultra-performance liquid chromatography/mass spectrometry analysis confirmed these results (Figure 1G–J). In vivo, tumour growth was effectively suppressed, as reflected by the significant reduction in tumour size and weight in the short hairpin RNA (shRNA) TRMT61A group. Immunohistochemistry staining showed a marked decrease in the proportion of tumour cells positively stained by anti-Ki-67 antibody in the shRNA TRMT61A group (Figure S1C and Figure 1K–M). Overall, TRMT61A promotes m1A modification and BLCA progression. To identify the target mRNAs of TRMT61A in BLCA, we conducted MeRIP-seq and RNA-seq analyses between control shRNA and TRMT61A shRNA. Gene Ontology and heatmap analysis revealed enrichment of genes involved in the regulation of mRNA processing and RNA splicing with decreased m1A methylation after TRMT61A knockdown; Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that transcripts with decreased m1A methylation were significantly related to the spliceosome and cancer pathways (Figure S2A–D). According to MeRIP-seq analysis, m1A peaks were particularly abundant in the vicinity of 5′ untranslated regions near coding sequence regions and were found throughout the genome and across all chromosomes (Figure 2A and Figure S2A). HMOX2 mRNA was evidently m1A de-methylated in the MeRIP-seq data and downregulated in RNA-seq data after TRMT61A knockdown, and overexpressed in human BLCA tissues; the same result was obtained with The Cancer Genome Atlas (TCGA) database (Figure 2B,C and Figure S2E). AGGCUGGA was the top m1A-modified motif; the HMOX2 mRNA m1A peak diminished upon TRMT61A knockdown (Figure S2F,G). Therefore, further investigations focused on HMOX2 as a TRMT61A target. We observed that HMOX2 was downregulated following TRMT61A knockdown in 5637 cells and upregulated in T24 cells overexpressing TRMT61A (Figure 2D). Using samples from BLCA patients, a positive correlation was observed between HMOX2 and TRMT61A expression; HMOX2 expression decreased in tumour tissues from mice xenografted with TRMT61A shRNA and increased in tumour tissues from BBN-driven urinary BLCA mice (Figures S2G–K and S3). TRMT61A promoted BLCA progression by upregulating the HMOX2 level (Figure S4). Overall, TRMT61A promotes BLCA progression by upregulating HMOX2 expression. Mechanistically, HMOX2 mRNA was enriched by the m1A antibody and m1A modification was reduced after TRMT61A knockdown in 5637 cells. RNA immunoprecipitation (RIP) analysis confirmed the interaction between TRMT61A protein and HMOX2 mRNA in 5637 cells. TRMT61A knockdown decreased the stability of HMOX2 mRNA (Figure 2E–G). YTHDF1 was screened through human BLCA tissues and the TCGA database (Figure 2H,I). RIP assays confirmed the interaction between YTHDF1 protein and HMOX2 mRNA; the interaction was attenuated upon TRMT61A knockdown. RNA pull-down assays confirmed that HMOX2 mRNA could directly bind to TRMT61A and YTHDF1 proteins. In mutant HMOX2, m1A modification was abrogated by the replacement of adenine with guanosine in the m1A consensus sequences. Luciferase assays revealed a significant increase in relative luciferase activity in cells transfected with the wild-type HMOX2 construct, but there was no significant change in luciferase activity in cells transfected with the mutant HMOX2 construct (Figure 2J–L). Next, we explored why TRMT61A is abnormally overexpressed in BLCA. Eleven transcriptional regulators were identified after intersecting transcription factors from the two prediction tools; nuclear factor kappa B (NF-κB) p56 was the transcriptional regulator most significantly associated with TRMT61A (Figure S5A). Real-time polymerase chain reaction (RT-PCR), western blot and IHC results showed that NF-κB was highly expressed in cancer tissues and positively correlated with TRMT61A expression (Figure 3A–C). Subsequently, 5637 cells were treated with SN50, an NF-κB inhibitor, which decreased p65 protein expression and phosphorylation (p-NF-κB, p-p65) and reduced TRMT61A protein expression. Conversely, p-p65 and TRMT61A levels increased after treatment with tumour necrosis factor-alpha (TNF-α), an NF-κB activator, in T24 cells (Figure 3D and Figure S5B,C). However, TRMT61A knockdown or overexpression did not affect p65 and p-p65 expression in 5637 and T24 cells (Figure S5D–F). Bioinformatics prediction by JASPAR revealed two NF-κB binding sites; chromatin immunoprecipitation confirmed strong binding of NF-κB at the -1105 to -1094 and -649 to -638 bp sites of the TRMT61A promoter. Experiments with dual-luciferase reporter genes confirmed the targeting relationship (Figure 3E–G). TRMT61A lacks a catalytic nuclear localization sequence.9 Thus, we examined whether NF-κB bound to TRMT61A protein and regulated its nuclear translocation. NF-κB activation facilitated the nuclear translocation of TRMT61A (Figure 3H–L). Finally, we identified TRMT61A inhibitors using a virtual screening strategy and two compounds (abbreviated as CMP) were potentially effective in inhibiting TRMT61A (Figure 4A,B, Figures S6–S8 and Table S1). Treatment with CMP1 or CMP9 significantly reduced m1A levels; RIP assays with anti-TRMT61A and anti-m1A antibodies and RT-qPCR analysis revealed that treatment with CMP1 or CMP9 significantly reduced TRMT61A protein binding to HMOX2 mRNA, HMOX2 mRNA m1A modification and HMOX2 mRNA expression (Figure 4C–F). In vivo, CMP1 and CMP9 showed strong anti-tumor effects, similar to DDP and Thiram (Figure 4G,H). CMP9 showed a good safety profile, while CMP1 increased serum Urea and Crea levels, indicating kidney toxicity exerted by CMP1 (Figure 4I,J). Consistent with the results of in vitro experiments, m1A levels and HMOX2 mRNA expression in tumour tissues decreased after CMP1 and CMP9 treatment (Figure 4K,L). Additional studies to demonstrate the anticancer efficacy of CMP9 in different BLCA mouse models and pharmacodynamics and pharmacokinetics studies are warranted to facilitate its potential applications. Overall, this study elucidates the molecular mechanisms via which NF-κB increases the transcription of TRMT61A, which accelerates m1A modification of HMOX2 mRNA and enhances its stability via a YTHDF1-dependent mechanism, ultimately promoting BLCA progression. The small molecule compound TRMT61A inhibitor CMP9 is a promising novel anticancer agent (Figure 4M). Jianjian Yin designed the study, performed experiments, analyzed and interpreted the data and wrote the manuscript. Dongkui Song, Lirong Zhang and Tao Liu conceived the study and reviewed and edited the manuscript. Fengmin Shao supervised the study and reviewed the manuscript. Xudong Zhang and Lei Jin supervised the experiments. Xin Fan, Qi Chang and Linlin Yang performed experiments and interpreted the data. Yuanheng Dai, Tao Wang, Lei Shi and Xiaoming Yang collected clinical specimens. All authors have read and approved the final manuscript. We thank Yongjie Lu (Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, UK) for the helpful discussions. The authors declare no conflict of interest. This work was funded by the National Natural Science Foundation of China (Grant Nos. U1904162 and 82373077). This study was approved by the Ethics Committee of the First Affiliated Hospital of Zhengzhou University, and written informed consent was obtained from the patients or their relatives prior to the study. All animal experiments were approved by the Zhengzhou University Animal Care and Ethics Committee (ZZUIRB2022-143). The original contributions presented in the study are included in the article/Supporting Information. The publicly available datasets analyzed in the current study are available from TCGA (https://www.cancer.gov/ccg/research/genome-sequencing/tcga). The RNA and MeRIP sequence datasets presented in this study can be found in online repositories (GSE255629). Further information and requests for resources and reagents should be directed to and will be fulfilled by the corresponding authors. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

OBJECTIVE:Retroperitoneal fibrosis is a rare disease characterized by chronic inflammation and fibrosis in the retroperitoneal space that may wrap around the ureter and cause an obstruction. Here we present the cases of three patients diagnosed and treated between April and August 2022. CASE PRESENTATION:Here we present three cases of retroperitoneal fibrosis. Imaging revealed hydronephrosis and/or ureteral dilation, whereas laboratory findings such as erythrocyte sedimentation rate, C-reactive protein, and immunoglobulin G4 were abnormal. Serum creatinine and blood urea nitrogen levels suggested renal injury. Additionally, two patients had unclear diagnoses from other hospitals. Later, all patients underwent ureteral stenting and laparoscopic ureterolysis at our hospital and simultaneous diagnosis by biopsy. The two patients with high immunoglobulin G4 levels exhibited extremely severe fibrosis. After discharge, each received maintenance prednisone to prevent recurrence as well as methotrexate, followed by follow-up every 3 months. Fortunately, serum creatinine, blood urea nitrogen, and intravenous urography findings remained normal with an average follow-up of 8.5 months. CONCLUSION:Here we discovered interesting findings in the diagnosis and treatment of retroperitoneal fibrosis, including the stable effect of medical plus surgical treatment, an unclear ureteral obstruction etiology, and the relationship between immunoglobulin G4 level and fibrosis hardness. However, the mechanisms behind these new findings require further study.

Bladder cancer (BCa) is the predominant malignancy of the urinary system. Herein, a comprehensive urine proteomic feature was initially established for the noninvasive diagnosis and recurrence monitoring of bladder cancer. 279 cases (63 primary BCa, 87 nontumor controls (NT), 73 relapsed BCa (BCR), and 56 nonrelapsed BCa (BCNR)) were collected to screen urinary protein biomarkers. 4761 and 3668 proteins were qualified and quantified by DDA and sequential window acquisition of all theoretical mass spectra (SWATH-MS) analysis in two discovery sets, respectively. Upregulated proteins were validated by multiple reaction monitoring (MRM) in two independent combined sets. Using the multi-support vector machine-recursive feature elimination (mSVM-RFE) algorithm, a model comprising 13 proteins exhibited good performance between BCa and NT with an AUC of 0.821 (95% CI: 0.675-0.967), 90.9% sensitivity (95% CI: 72.7-100%), and 73.3% specificity (95% CI: 53.3-93.3%) in the diagnosis test set. Meanwhile, an 11-marker classifier significantly distinguished BCR from BCNR with 75.0% sensitivity (95% CI: 50.0-100%), 81.8% specificity (95% CI: 54.5-100%), and an AUC of 0.784 (95% CI: 0.609-0.959) in the test cohort for relapse surveillance. Notably, six proteins (SPR, AK1, CD2AP, ADGRF1, GMPS, and C8A) of 24 markers were newly reported. This paper reveals novel urinary protein biomarkers for BCa and offers new theoretical insights into the pathogenesis of bladder cancer (data identifier PXD044896).

Although immune checkpoint blockade therapy (ICBT) has revolutionized cancer treatment with good therapeutic response in a number of human cancers, including bladder cancer, many cancers still do not respond to ICBT. Analyzing genetic signatures helps the understanding of underlying biological mechanisms. Here, based on two cohorts of bladder cancer patients receiving ICBT, we identified three novel ICBT-associated signatures in the bladder cancer microenvironment, involving genomic stability, angiogenesis and RNA regulatory, which affect PD-L1 expression and patient response to ICBT. The combinations of these signatures with TMB or PD-L1 expression improved the overall survival prediction efficiency over TMB and PD-L1 expression alone for patients receiving ICBT. Moreover, we utilized two methods to search potential drugs or small-molecules that have an impact on ICBT-associated signatures. This study provides new molecular insight into ICBT response of bladder cancer and has the potential to improve the prediction accuracy for patients to benefit from ICBT.

Introduction: Post-transcriptional RNA modifications are crucial regulators of tumor development and progression. In many biological processes, N1-methyladenosine (m1A) plays a key role. However, little is known about the links between chemical modifications of messenger RNAs (mRNAs) and long noncoding RNAs (lncRNAs) and their function in bladder cancer (BLCA).Methods: Methylated RNA immunoprecipitation sequencing and RNA sequencing were performed to profile mRNA and lncRNA m1A methylation and expression in BLCA cells, with or without stable knockdown of the m1A methyltransferase tRNA methyltransferase 61A (TRMT61A).Results: The analysis of differentially methylated gene sites identified 16,941 peaks, 6,698 mRNAs, and 10,243 lncRNAs in the two groups. Gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the differentially methylated and expressed transcripts showed that m1A-regulated transcripts were mainly related to protein binding and signaling pathways in cancer. In addition, the differentially genes were identified that were also differentially m1A-modified and identified 14 mRNAs and 19 lncRNAs. Next, these mRNAs and lncRNAs were used to construct a lncRNA-microRNA-mRNA competing endogenous RNA network, which included 118 miRNAs, 15 lncRNAs, and 8 mRNAs. Finally, the m1A-modified transcripts, SCN2B and ENST00000536140, which are highly expressed in BLCA tissues, were associated with decreased overall patient survival.Discussion: This study revealed substantially different amounts and distributions of m1A in BLCA after TRMT61A knockdown and predicted cellular functions in which m1A may be involved, providing evidence that implicates m1A mRNA and lncRNA epitranscriptomic regulation in BLCA tumorigenesis and progression.

One of the most abundant protein-protein interaction domains in the human proteome is the WD40 repeat (WDR) domain. A Gene Expression Omnibus dataset revealed 37 differentially expressed WDR domain genes in bladder cancer (BC). WD repeat domain 54 (WDR54), an upregulated WDR domain gene, was selected for further investigation. Sixty pairs of frozen BC tumor and non-malignant bladder tissues and 83 paraffin-embedded BC tissue specimens were obtained. Loss-/gain-of-function experiments were carried out using BC and xenograft tumor models. WDR54 was overexpressed in BC cells, and its high expression was linked to tumor stage and lymph node metastases in patients. WDR54 contributed to the tumorigenesis and metastasis of BC and impaired its chemosensitivity. WDR54 prevented the degradation and ubiquitination of the mediator of ErbB2-driven cell motility 1 (MEMO1). WDR54 also promoted the interaction between MEMO1 and insulin receptor substrate 1 (IRS1) and activated the IRS1/AKT/β-catenin pathway in BC cells. Particularly, WDR54 depended on MEMO1 to exert its biological functions. Our study demonstrated the relevance of WDR54 in BC and provides insight into the molecular mechanism underlying BC.

目的 基于全身炎症指标和临床特征探讨影响腺性膀胱炎(CG)电切术后复发的危险因素.方法 回顾性分析2016-01-2021-10 就诊于郑州大学第一附属医院的CG患者,按照膀胱镜下表现分为高危型CG及低危型CG.以无复发生存期(RFS)为结局指标,将中性粒细胞与淋巴细胞比值(NLR)、血小板与淋巴细胞比值(PLR)、全身免疫炎症指数(SII)的全身炎症指标的中位数作为截点,分为高危组和低危组.在中位生存期25 个月(3～56 个月)的随访中,依据CG是否复发,分为复发组与未复发组.以Cox风险回归模型对全身炎症指标和临床特征进行单因素及多因素分析,探讨影响 CG及高危型 CG术后复发的危险因素.结果 NLR、SII、临床分型、膀胱热灌注、性别、年龄与CG术后复发相关联(P＜0.05).将单因素分析筛选的因素纳入多因素分析显示,临床分型、膀胱热灌注和NLR是CG术后复发的独立危险因素(P＜0.05).膀胱热灌注、NLR、SII与高危型CG术后复发相关联(P＜0.05).将单因素分析筛选的因素纳入多因素分析显示,膀胱热灌注和NLR是高危型CG复发的独立危险因素.结论 临床分型、膀胱热灌注、NLR可作为CG术后复发的独立预测因素.高危型CG膀胱热灌注治疗可显著减少术后复发;低危型CG膀胱热灌注治疗不能使患者显著获益.

目的 探讨长基因间非编码RNA(LINC)02163 靶向微小RNA(miR)-582-3p对膀胱癌进展的影响.方法实时定量聚合酶链反应(RT-qPCR)分析 23 例膀胱癌组织和对应癌旁组织中 LINC02163 和 miR-582-3p 的表达水平.将LINC0216 小干扰RNA(si-LINC02163)、miR-582-3p模拟物、si-LINC02163+miR-582-3p抑制物(anti-miR-582-3p)分别转染膀胱癌T24 细胞,通过噻唑蓝(MTT)、克隆形成、伤口愈合和Transwell实验检测LINC02163 和miR-582-3p表达对T24 生物学行为的影响.Western印迹测定上皮型钙黏蛋白(E-cadherin)和神经型钙黏蛋白(N-cadherin)蛋白表达量.RT-qPCR和双荧光素酶报告实验用于确定LINC02163 和miR-582-3p靶向关系.结果 与癌旁组织比较,膀胱癌组织中LINC02163 表达量显著升高miR-582-3p表达量显著降低(P<0.05).沉默LINC02163 显著降低T24 细胞活力、划痕愈合率和N-cadherin表达量显著减少集落形成数和侵袭数,显著增加凋亡率和E-cadherin表达量(P<0.05).过表达miR-582-3p显著降低T24 细胞活力、划痕愈合率和N-cadherin表达量,显著减少集落形成数和侵袭数,显著增加凋亡率和E-cadherin表达量(P<0.05).LINC02163 与miR-582-3p直接结合,沉默 LINC02163 显著增加 miR-582-3p 表达量(P<0.05).抑制 miR-582-3p 表达显著削弱沉默LINC02163 对T24 细胞生物学行为和 E-cadherin、N-cadherin 表达的影响.结论 膀胱癌中 LINC02163 表达上调,沉默LINC02163 通过靶向抑制miR-582-3p表达可抑制膀胱癌细胞增殖、迁移和侵袭,诱导细胞凋亡,抑制膀胱癌进展.

Penile cancer is a rare malignancy with poor prognosis. Advanced penile cancer that invades the penile shaft and causes urinary retention could be rarely seen. We reported a 72-year-old male suffered from advanced penile cancer with penile self-amputation and acute urinary retention. We reckoned the self-amputation of penis as a new sign of advanced penile cancer.

目的 探讨影响晚期前列腺癌内分泌治疗患者预后的危险因素,并以此建立一个生存风险分层模型.方法 本研究纳入2013-01-2020-12 在郑州大学第一附属医院泌尿外科以内分泌治疗为主要治疗手段的 157 例IV期前列腺癌患者.收集患者的功能状态(PS)、血小板计数(PLT)、前列腺特异性抗原(PSA)、碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)、Gleason评分(GS)、血红蛋白(Hb)、疾病程度(EOD)和腺癌分化高低等数据信息.使用Kaplan-Meier方法评估疾病特异性生存(DSS),确定与DSS相关的指标并使用Cox比例风险模型多因素分析得出独立的预后因素.计算DSS中独立预后因素的危险率(HR),根据HR将患者分为高风险、中风险、低风险3 组(确定3 组间DSS有统计学意义),以此建立了生存风险分层模型.结果 单因素分析中,PS、PLT、PSA、ALP、LDH、GS评分,Hb、EOD和腺癌分化高低,是疾病特异性生存(DSS)的影响因素,差异有统计学意义(P＜0.05).在多变量Cox比例风险模型中,PS和PLT是独立的预后因素.根据危险率(HR)将患者分为 3 个危险组,3 组间 DSS 率差异有统计学意义(P＜0.05).结论 这些结果表明,PS和PLT是晚期前列腺癌内分泌治疗独立的预后因素,联合两个因素可以根据DSS风险对接受内分泌治疗的晚期前列腺癌患者进行分层并建立生存风险分层模型.

Abstract Background: Bladder cancer is one of the most common urinary tract tumors. At present, for the treatment of invasive muscular bladder cancer or high-risk non-invasive muscular bladder cancer, radical cystectomy, urinary diversion and pelvic lymph node dissection are the preferred methods. Ureterostomy is one of the commonly used surgical procedures for postoperative urinary diversion. Open radical cystectomy (ORC), laparoscopic radical cystectomy (LRC) and robot-assisted laparoscopic radical cystectomy (RARC) are common radical cystectomy methods commonly used by urologists. Methods: In this study, patients with bladder cancer were treated with 3 different surgical methods, and the postoperative efficacy was compared, and the survival difference was analyzed. Results: Our results showed that LRC and RARC were significantly superior to ORC in intraoperative and perioperative observation indexes, and LRC and RARC could more completely remove tumors compared with ORC in complex cases. However, the survival time after ORC was longer than that of LRC and RARC patients. Conclusion: Operation methods, tumor staging, pathological grade, and number of lymph nodes dissected are all independent factors affecting patient survival, which should be paid more attention to clinically.

Purpose: Bladder cancer is a kind of common malignant cancer in the urinary system. The expression of EDARADD (ectodysplasin-A receptor-associated death domain) in bladder cancer is higher than the normal samples. However, its role in bladder cancer remains unknown. In the present study, we analyzed the expression of EDARADD in 81 bladder cancer samples by immunohistochemistry as well as its correlation with clinical characteristics. In addition, the role of EDARADD was also explored through loss of function. Materials and Methods: Cell proliferation assay and MTT assay were conducted to assess the proliferation of bladder cancer cells and transwell assay and wound healing assay were conducted to assess the migration of bladder cancer cells. On the other hand, the levels of epi-thelial-mesenchymal transition (EMT) associated proteins and the key molecules in the MAPK signaling pathway were detected by western blot. In vivo experiments were also conducted to determine the effect of EDARADD silencing on the metastasis of bladder cancer cells and the MAPK signaling pathway. Results: EDARADD was highly expressed in bladder cancer samples, especially in high-grade bladder cancer samples. The high EDARADD level indicated a poor survival. Interestingly, EDARADD silencing suppressed the proliferation, migration and EMT of bladder cancer cells. Furthermore, the MAPK signaling pathway was repressed by EDARADD silencing. Additionally, silencing EDARADD also inhibited the metastasis of bladder cancer and the MAPK signaling pathway in vivo. It is indicated that silencing EDARADD may suppress the proliferation and metastasis of bladder cancer cells through the MAPK signaling pathway. Conclusion: These results indicate that EDARADD may become a probable target for the treatment of bladder cancer. (c) 2022 Elsevier Inc. All rights reserved.

The exploration of nucleoside changes in human biofluids has profound potential for cancer diagnosis. Herein, we developed a rapid methodology to quantify 17 nucleosides by UHPLC-MS/MS. Five pairs of isomers were successfully separated within 8 min. The ME was mostly eliminated by sample dilution folds of 1000 for urine and 40 for CCS. The optimized method was firstly applied to screen potential nucleoside biomarkers in CCS by comprising bladder cancer cell lines (5637 and T24) and normal human bladder cell line SV-HUC-1 together with student's t-test and OPLS-DA. Nucleosides with significant differences in the supernatant of urine samples were also uncovered comparing BCa with the non-tumor group, as well as a comparison of BCa recurrence group with the non-recurrence group. By intersecting the differential nucleosides in CCS and urine supernatant, and then further confirmed using validation sets, the combination of m3C and m1A with AUC of 0.775 was considered as a potential biomarker for bladder cancer diagnosis. A panel of m3C, m1A, m1G, and m22G was defined as potential biomarkers for bladder cancer prognosis with an AUC of 0.819. Above all, this method provided a new perspective for diagnosis and recurrence monitoring of bladder cancer.Significance: The exploration of nucleoside changes in body fluids has profound potential for the diagnosis and elucidation of the pathogenesis of cancer. In this study, we developed a rapid methodology for the simultaneous quantitative determination of 17 nucleosides in the supernatant of cells and urine samples using UHPLC-MS/MS to discover and validate bladder cancer related excreted nucleoside biomarkers. The results of this paper provide a new strategy for diagnosis and postoperative recurrence monitoring of bladder cancer and provide theoretical support for the exploration of its pathogenesis.

Background: The emergence of castration resistance is fatal for patients with prostate cancer (PCa); however, there is still a lack of effective means to detect the early progression. In this study, a novel combined nomogram was established to predict the risk of progression related to castration resistance.Methods: The castration-resistant prostate cancer (CRPC)-related differentially expressed genes (DEGs) were identified by R packages "limma" and "WGCNA" in GSE35988-GPL6480 and GSE70768-GPL10558, respectively. Relationships between DEGs and progression-free interval (PFI) were analyzed using the Kaplan-Meier method in TCGA PCa patients. A multigene signature was built by lasso-penalized Cox regression analysis, and assessed by the receiver operator characteristic (ROC) curve and Kaplan-Meier curve. Finally, the univariate and multivariate Cox regression analyses were used to establish a combined nomogram. The prognostic value of the nomogram was validated by concordance index (C-index), calibration plots, ROC curve, and decision curve analysis (DCA).Results: 15 CRPC-related DEGs were identified finally, of which 13 genes were significantly associated with PFI and used as the candidate genes for modeling. A two-gene (KIFC2 and BCAS1) signature was built to predict the risk of progression. The ROC curve indicated that 5-year area under curve (AUC) in the training, testing, and whole TCGA dataset was 0.722, 0.739, and 0.731, respectively. Patients with high-risk scores were significantly associated with poorer PFI (p < 0.0001). A novel combined nomogram was successfully established for individualized prediction integrating with T stage, Gleason score, and risk score. While the 1-year, 3-year, and 5-year AUC were 0.76, 0.761, and 0.762, respectively, the good prognostic value of the nomogram was also validated by the C-index (0.734), calibration plots, and DCA.Conclusion: The combined nomogram can be used to predict the individualized risk of progression related to castration resistance for PCa patients and has been preliminarily verified to have good predictive ability.

目的 分析表面活性蛋白A(SP-A)和表面活性蛋白D(SP-D)基因多态性及尿液中含量与膀胱癌术后尿路感染的关联性.方法 选择2019年1月-2020年1月于新乡医学院第一附属医院行经尿道膀胱肿瘤切除术的203例膀胱癌患者,分为术后尿路感染组(n=38)和无尿路感染组(n=165),采用聚合酶链反应-序列特异性引物(PCR-SSP)分析SP-A及SP-D基因位点单核苷酸多态性(SNPs),比较两组尿液中SP-A及SP-D含量.结果 尿路感染组SP-A1基因Ala19 Val位点Val/Val基因型频率、19 Val等位基因频率分别为15.79％、48.68％,SP-A2基因Lys223Gln位点Lys/Lys基因型频率、223Lys等位基因频率分别为5.26％、14.47％,SP-D基因Thr11 Met位点Met/Met基因型频率、11 Met等位基因频率分别为26.32％、53.95％,均低于无尿路感染组(P<0.05);尿路感染组尿液中SP-A及SP-D含量均低于无尿路感染组(P<0.05).结论 SP-A1基因Ala19Val位点19Val等位基因、SP-A2基因Lys223Gln位点223Gln等位基因以及SP-D基因Thr11Met位点11 Met等位基因突变,以及尿液中SP-A及SP-D含量降低可能与膀胱癌术后尿路感染存在关联性.

Bladder cancer (BC) is the most common type of malignant tumor in the genitourinary system. Through the microarray analysis of clinical samples, long noncoding RNA HAND2-AS1 expression was found to be downregulated in BC tissues. However, the function of HAND2-AS1 on BC and underlying mechanism are unclear. In this study, the correlations of HAND2-AS1 with clinicopathological parameters in BC patients were determined. The gain- and loss-of-function experiments were conducted to examine the role of HAND2-AS1 in malignant behaviors of BC cells in vitro and in vivo. Then, we paid attention to miR-17-5p/KLF9 axis to illustrate the molecular mechanism. Results showed that HAND2-AS1 was downregulated in BC tissues, and its overexpression significantly inhibited cell proliferation, migration, and invasion in vitro, as well as tumor growth in vivo. Knockdown of HAND2-AS1 caused an opposite effect on BC cell malignancies. Furthermore, miR-17-5p was shown to be a direct target of HAND2-AS1, and it reversed the inhibitory effect of HAND2-AS1 on BC malignancies. Also, as a downstream factor of miR-17-5p, KLF9 silencing was demonstrated to mediate the role of miR-17-5p inhibitor in BC cell proliferation and invasion. Thus, it suggests that HAND2-AS1 acts as a suppressor in BC development through miR-17-5p/KLF9 axis.

目的:研究早期有丝分裂抑制因子1(EMI1)在膀胱癌组织中的表达及其与患者临床病理特征和预后的关系.方法:选取经尿道膀胱肿瘤电切术或膀胱癌根治术切除的膀胱癌组织112例,及其配对癌旁正常膀胱组织86例,采用免疫组织化学SP法检测两种组织中EMI1蛋白的表达.分析EMI1表达与膀胱癌患者临床病理特征的关系,采用Kaplan-Meier生存曲线比较EMI1高表达组和低表达组膀胱癌患者的生存差异,采用Cox回归分析EMI1的表达水平与患者预后的关系.结果:膀胱癌组织和配对癌旁正常组织中EMI1高表达率分别为46.5％和25.6％,差异有统计学意义(P<0.05).EMI1蛋白表达水平与膀胱癌组织类型、T分期、TNM分期、病理分级有关(P<0.05).EMI1高表达组和低表达组生存曲线差异有统计学意义(P<0.05).远处转移、TNM分期Ⅲ+Ⅳ期、EMI1高表达是膀胱癌预后的危险因素[HR(95％CI)分别为3.891(1.350～11.215)、2.626(1.117～6.174)、3.358(1.378～8.158)].结论:EMI1在膀胱癌组织中表达升高,且表达水平与膀胱癌组织病理分级、临床分期有关,对患者预后的判断有一定指导意义.

目的 探讨术前预后营养指数(PNI)和中性粒细胞与淋巴细胞比值(NLR)对非肌层浸润性膀胱癌(NMIBC)患者预后的影响.方法 回顾性分析202例2013年1月至2017年3月在郑州大学第一附属医院初诊为NMIBC患者的临床资料.采用受试者工作特征(ROC)曲线确定PNI和NLR的最佳临界值,采用Kaplan-Meier法进行生存分析,比较患者的无复发生存时间(RFS),并采用Cox回归模型确定影响NMIBC患者预后的独立危险因素.结果 根据受试者工作特征曲线(ROC),按PNI<48.5(67例)及≥48.5(135例);NLR<2.20(139例)及≥2.20(63例)分组.高PNI组患者的年龄、性别、吸烟史、组织学分级和病理T分期与低PNI组差异有统计学意义(P<0.05).高NLR组患者的肿瘤数量、肿瘤最大径、组织学分级和病理T分期与低NLR组差异有统计学意义(P<0.05).高PNI组患者无复发生存时间大于低PNI组,低NLR组患者无复发生存时间大于高NLR组,差异有统计学意义(P<0.05).单因素分析结果显示年龄、吸烟史、肿瘤最大径、肿瘤数量、病理T分期、组织学分级、PNI和NLR与NMIBC患者的RFS密切相关(P<0.05).多因素Cox分析结果显示病理T分期、PNI和NLR是影响NMIBC患者预后的独立危险因素.结论 PNI和NLR是影响患者无复发生存率的独立危险因素,并可以作为评估NMIBC患者预后的指标.联合PNI和NLR能够增加预后评估的精确性.