Intraoperative blood loss (IBL) is a critical precipitating factor of intraoperative and postoperative complications. This study aims to identify risk factors and employ machine learning (ML) techniques to develop models for predicting IBL in patients undergo retroperitoneal laparoscopic adrenalectomy (RLA). A retrospective study was performed with data from patients who underwent unilateral RLA at Beijing Anzhen Hospital from 2014 to 2021. The data were randomly divided into training and validation sets. A volume ≥ 100 ml was defined as high IBL. Model training incorporated feature selection with least absolute shrinkage and selection operator (LASSO) and algorithms including logistic regression (LR), random forest (RF), neural network (NN), K-nearest neighbor (KNN), support vector machine (SVM) and AdaBoost. Various indicators were utilized to evaluate model performance. A total of 530 RLA cases were included. LASSO regression identified gender, surgeons’ experience, disease type, lesion diameter and lesion location as predictive factors. Six ML models were derived and evaluated. Among which, LR, RF and NN models exhibited exceptional discriminatory power. The area under the curve (AUC) of the three models were 0.775, 0.769, 0.729 in the validation set, respectively. A nomogram and a online calculator were created. The calibration curve, decision curve analysis and clinical impact curve revealed its excellent clinical utility. This study presented and compared the use of various ML models for predicting IBL in RLA, which could assist in assessing surgical risks and tailored treatments without additional examination, and providing insights for the clinical application of ML.

It is unclear how sex disparities affect surgical outcomes following retroperitoneal laparoscopic adrenalectomy (RLA). Therefore, we studied whether gender could affect RLA outcomes perioperatively. A retrospective study was performed involving consecutive cases underwent unilateral RLA for adrenal disease from January 2012 to December 2021. Coarsened exact matching (CEM) was applied to balance patient groups. Logistic regression analysis was performed to assess the impact of sex on prolonged operative time and increased intraoperative blood loss. A total of 569 patients who underwent RLA were included. After CEM, 112 patients were incorporated in each group. Operative time (male: 129.47 vs. female: 95.53, P < 0.001) and intraoperative blood loss (male: 123.39 vs. female: 101.61, P = 0.009) differed statistically significantly, while equal postoperative length of stay (LOS) (6.7 d vs. 6.8 d, P = 0.686), drainage tube removal time (3.0 d vs. 3.0 d, P = 0.231) and incidence of Clavien-Dindo grade ≥ 2 perioperative complications (8.0

BACKGROUND:Thrombosis was recognized as a significant cause of morbidity and mortality for prostate cancer (PCa). The potential mechanisms underlying the effect of thrombosis on PCa remain elusive. METHODS:Retrospective analysis of dual-center clinical data identified thrombosis-PCa correlations. Bioinformatics integration of TCGA-PRAD transcriptomics and thrombosis-related genes enabled construction of a prognostic signature, validated externally. Functional, immune, and drug sensitivity analyses were performed. Experimental validation included IHC, qPCR, IF, in vitro functional assays, and in vivo models. RESULTS:Elevated thrombosis risk strongly correlated with PCa aggressiveness and adverse clinical features. A five-gene risk model stratified PCa patients into distinct survival groups (low-risk: superior outcomes), validated by ROC/Cox analyses as an independent prognostic tool. Findings from functional enrichment, alongside evaluations of immune cell infiltration, immunotherapy responsiveness, and drug sensitivity, reinforced the capacity to accurately forecast the clinical efficacy of precision therapies, as validated by clinical relevance analysis and nomogram development. Immunohistochemistry and qPCR of signature genes revealed marked differences between PCa and adjacent tissues. Importantly, experimental knockdown of VAV2 in 22RV1 cells downregulated prothrombotic inflammatory factors (CXCL8, IL-6, and VEGF). Conditioned media from VAV2-knockdown cells markedly inhibited tube formation in HUVECs and suppressed NETs formation. In vivo, mice administered with conditioned media from VAV2-deficient cells exhibited prolonged PT and APTT, and reduced fibrinogen levels, indicating attenuated coagulation potential. CONCLUSION:We successfully developed and validated an innovative and robust five-gene signature, seamlessly integrating clinical prognostic parameters for the precise prediction of PCa patients outcomes. This dissertation offers an in-depth exploration of thrombosis, elucidating potential biological mechanisms underpinning therapeutic strategies related to tumor immunity in PCa.

Figure S2 showed the increased GPD1 expression was seen in cancer patients in TCGA database and GPD1 overexpression enhanced the anti-proliferation ability of metformin in vitro.

Figure S3 showed the detection of G3P, DHAP and methylglyoxal in cancer cells treated with or without different concentrations of G3P.

BackgroundClear cell renal cell carcinoma (ccRCC) is a malignant disease containing tumor-infiltrating lymphocytes. Reactive oxygen species (ROS) are present in the tumor microenvironment and are strongly associated with cancer development. Nevertheless, the role of ROS-related genes in ccRCC remains unclear. MethodsWe describe the expression patterns of ROS-related genes in ccRCC from The Cancer Genome Atlas and their alterations in genetics and transcription. An ROS-related gene signature was constructed and verified in three datasets and immunohistochemical staining (IHC) analysis. The immune characteristics of the two risk groups divided by the signature were clarified. The sensitivity to immunotherapy and targeted therapy was investigated. ResultsOur signature was constructed on the basis of glutamate-cysteine ligase modifier subunit (GCLM), interaction protein for cytohesin exchange factors 1 (ICEF1), methionine sulfoxide reductase A (MsrA), and strawberry notch homolog 2 (SBNO2) genes. More importantly, protein expression levels of GCLM, MsrA, and SBNO2 were detected by IHC in our own ccRCC samples. The high-risk group of patients with ccRCC suffered lower overall survival rates. As an independent predictor of prognosis, our signature exhibited a strong association with clinicopathological features. An accurate nomogram for improving the clinical applicability of our signature was constructed. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that the signature was closely related to immune response, immune activation, and immune pathways. The comprehensive results revealed that the high-risk group was associated with high infiltration of regulatory T cells and CD8+ T cells and more benefited from targeted therapy. In addition, immunotherapy had better therapeutic effects in the high-risk group. ConclusionOur signature paved the way for assessing prognosis and developing more effective strategies of immunotherapy and targeted therapy in ccRCC.

Objective:To explore the risk factors of prolonged operation time and higher intraoperative blood loss of retroperitoneal laparoscopic radical nephrectomy (LRN).Methods:The data of patients who underwent retroperitoneal LRN from August 2010 to December 2021 were analyzed retrospectively. Demographic and hospital admission data were collected from these patients with complete medical records. Furthermore, univariate and multivariate logistic analysis were performed to determine the risk factor related to prolonged operation time and increased blood lose following LRN. Receiver operating characteristic (ROC) curve was used to assess the value of risk factors for predicting prolonged operation time and high intraoperative blood loss.Results:A total of 22 patients (24.4%) had prolonged operation time, and 14 patients (15.6%) had high intraoperative blood loss. The risk factor associated with prolonged operation time in a multivariate analysis was surgical experience (OR=0.13, P<0.001). The area under the ROC curve was 0.759 (95% confidence interval: 0.638-0.880) in the multivariate logistic regression model for predict prolonged operation time. Those associated with high intraoperative blood loss were surgical experience (OR=0.25, P=0.032) and tumor T stage (OR=3.18, P=0.007). The area under the ROC curve was 0.769 (95% confidence interval: 0.627-0.910) in the multivariate logistic regression model for predict high intraoperative blood loss.Conclusions:Surgical experience and tumor T stage were associated with the risk of prolonged operation time and high intraoperative blood loss.

Figure S5 showed that metformin can suppress OCR through inhibiting GPD2 expression.

Abstract Background Immune checkpoint inhibitors (ICIs) have demonstrated promising outcomes in small cell lung cancer (SCLC), but not all patients benefit from it. Thus, developing precise treatments for SCLC is a particularly urgent need. In our study, we constructed a novel phenotype for SCLC based on immune signatures. Methods We clustered patients with SCLC hierarchically in 3 publicly available datasets according to the immune signatures. ESTIMATE and CIBERSORT algorithm were used to evaluate the components of the tumor microenvironment. Moreover, we identified potential mRNA vaccine antigens for patients with SCLC, and qRT-PCR were performed to detect the gene expression. Results We identified 2 SCLC subtypes and named Immunity High (Immunity_H) and Immunity Low (Immunity_L). Meanwhile, we obtained generally consistent results by analyzing different datasets, suggesting that this classification was reliable. Immunity_H contained the higher number of immune cells and a better prognosis compared to Immunity_L. Gene-set enrichment analysis revealed that several immune-related pathways such as cytokine-cytokine receptor interaction, programmed cell death-Ligand 1 expression and programmed cell death-1 checkpoint pathway in cancer were hyperactivated in the Immunity_H. However, most of the pathways enriched in the Immunity_L were not associated with immunity. Furthermore, we identified 5 potential mRNA vaccine antigens of SCLC (NEK2, NOL4, RALYL, SH3GL2, and ZIC2), and they were expressed higher in Immunity_L, it indicated that Immunity_L maybe more suitable for tumor vaccine development. Conclusions SCLC can be divided into Immunity_H and Immunity_L subtypes. Immunity_H may be more suitable for treatment with ICIs. NEK2, NOL4, RALYL, SH3GL2, and ZIC2 may be act as potential antigens for SCLC.

Supplementary Information showed the additional details, such as reagents and assay kit, list of antibodies, sequences, primers and abbreviation.

Background: While laparoscopic adrenalectomy (LA) represents a gold standard for treating most adrenal lesions, no effective visual model for the prediction of perioperative complications of retroperitoneal laparoscopic adrenalectomy (RLA) exists.Methods: A retrospectively study was conducted involving all consecutive patients underwent unilateral RLA for adrenal disease from January 2012 to December 2021. The entire cohort was randomly divided into 2 subsets (70% of the data for training, 30% for validation). Subsequently, a Least Absolute Shrinkage Selection Operator (LASSO) regression was performed to select the predictor variables, which were further consolidated via random forest (RF) and Boruta algorithm. Then the nomogram was established using the bivariate logistic regression analysis. Eventually, the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA) were employed to evaluate discrimination, calibration and clinical usefulness of the model, respectively.Results: A total of 610 patients underwent unilateral RLA for adrenal diseases were enrolled. After machine learning analyses, a weighted nomogram was established with 7 factors associated with complications including operative time, lesion laterality, intraoperative blood loss, pheochromocytoma, body mass index (BMI) and 2 preoperative comorbidities (respiratory diseases, cardiovascular diseases (CVD)). The model displayed a fine calibration curve for perioperative complications evaluation in both the training dataset (P=0.847) and validation dataset (P=0.248). ROC with AUC revealed an excellent discrimination in the training dataset (0.817, 95% CI: 0.758-0.875) and validation dataset (0.794, 95% CI: 0.686-0.901). DCA curves showed that using this nomogram provided a more net benefit where threshold probabilities lay in the range of 0.1 to 0.9.Conclusions: An effective nomogram that incorporating 7 predictors was established in this study to identify patients at high risk of perioperative complications for RLA. It would contribute to the improvement of perioperative strategy due to its accuracy and convenience.

Background Hepatocellular carcinoma (HCC) is a complex disease with a poor outlook for patients in advanced stages. Immune cells play an important role in the progression of HCC. The metabolism of sphingolipids functions in both tumor growth and immune infiltration. However, little research has focused on using sphingolipid factors to predict HCC prognosis. This study aimed to identify the key sphingolipids genes (SPGs) in HCC and develop a reliable prognostic model based on these genes. Methods The TCGA, GEO, and ICGC datasets were grouped using SPGs obtained from the InnateDB portal. A prognostic gene signature was created by applying LASSO-Cox analysis and evaluating it with Cox regression. The validity of the signature was verified using ICGC and GEO datasets. The tumor microenvironment (TME) was examined using ESTIMATE and CIBERSORT, and potential therapeutic targets were identified through machine learning. Single-cell sequencing was used to examine the distribution of signature genes in cells within the TME. Cell viability and migration were tested to confirm the role of the key SPGs. Results We identified 28 SPGs that have an impact on survival. Using clinicopathological features and 6 genes, we developed a nomogram for HCC. The high- and low-risk groups were found to have distinct immune characteristics and response to drugs. Unlike CD8 T cells, M0 and M2 macrophages were found to be highly infiltrated in the TME of the high-risk subgroup. High levels of SPGs were found to be a good indicator of response to immunotherapy. In cell function experiments, SMPD2 and CSTA were found to enhance survival and migration of Huh7 cells, while silencing these genes increased the sensitivity of Huh7 cells to lapatinib. Conclusion The study presents a six-gene signature and a nomogram that can aid clinicians in choosing personalized treatments for HCC patients. Furthermore, it uncovers the connection between sphingolipid-related genes and the immune microenvironment, offering a novel approach for immunotherapy. By focusing on crucial sphingolipid genes like SMPD2 and CSTA, the efficacy of anti-tumor therapy can be increased in HCC cells.

Figure S4 showed that the extracellular acidification rate assay in GPD1 overexpressing cells and the CCK8 assay and the oxygen consumption rate in GPD1 knock-down cells.

目的 探讨经腹膜后入路腹腔镜肾上腺切除术(RLA)手术时间延长的危险因素.方法 回顾性分析2016年1月-2021年12月该院泌尿外科收治的420例因肾上腺病变行RLA手术的患者的临床资料,包括:年龄、性别、体重指数(BMI)、合并症、既往腹部手术史、手术时间、瘤体大小、术后病理类型和肿瘤位置等.以手术时间的第75百分位数(140 min)为分界点,手术时间超过140 min的,定义为手术时间延长.分别应用单因素和多因素Logistic回归模型,分析引起RLA手术时间延长的危险因素,计算受试者操作特征曲线(ROC curve)的曲线下面积(AUC),分析相关因素对RLA手术时间延长的预测价值.结果96例(22.86%)出现手术时间延长.单因素Logistic回归分析结果显示,男性、BMI≥31 kg/m2、术者经验(≤30例)、手术方式(肾上腺全切)、瘤体直径≥3.6 cm、病理类型(肾上腺皮质增生和嗜铬细胞瘤),与RLA手术时间延长有关.多因素Logistic回归分析结果显示,男性、BMI≥31 kg/m2、术者经验(≤30例)、瘤体直径≥3.6 cm和嗜铬细胞瘤,是引起RLA手术时间延长的独立危险因素.以病理类型(嗜铬细胞瘤)、性别(男性)、术者经验(≤30 例)、BMI≥31 kg/m2 和瘤体直径≥3.6 cm为预测因素,AUC为 0.735(95%CI:0.678～0.793).结论 RLA手术时间延长与性别、BMI、病理类型、术者经验和瘤体直径等因素有关,术前对这些危险因素进行识别,有助于更准确地选择手术方案,缩短手术时间.

Figure S6 showed that metformin induced AMPK phosphorylation and inhibited mTOR pathway, which were not affected by GPD1 overexpression.

The study aimed to investigate the anti-tumor effects and underlying mechanisms of Enzalutamide (ENZ) and Arsenic trioxide (ATO) co-treatment on castration-resistant prostate cancer (CRPC). The effects on C4-2B cells were initially evaluated by colony formation assay, FACS analysis, and DNA fragmentation detection. Bioinformatics methods including mRNA-sequencing and gene enrichment analysis were used to screen the underlying target genes and pathways related to their actions. Western blot was employed to assess the expression levels of protein-related angiogenesis, apoptosis, DNA repair, and the screened genes. Finally, the effects were further verified in subcutaneous tumor models and tissue sections from the xenografts. It was found that not only could ENZ combination with ATO significantly inhibit cell proliferation and angiogenesis, but also induce cell arrest and apoptosis in C4-2B cells. In addition, interruption of the DNA damage repair-related pathways also occurred as a result of their combined effects. Western blot analysis further suggested that proteins involved in these pathways, especially P-ATR and P-CHEK1 were significantly reduced. In addition, their combination also inhibited the tumor growth of xenografts. Altogether, ENZ combination with ATO synergistically improved the therapeutic effects and suppressed CRPC progression through regulation of the ATR-CHEK1-CDC25C pathway.

Voltage-gated chloride ion channels (CLCs) are transmembrane proteins that maintain chloride ion homeostasis in various cells. Accumulating studies indicated CLCs were related to cell growth, proliferation, and cell cycle. Nevertheless, the role of CLCs in prostate cancer (PCa) has not been systematically profiled. The purpose of this study was to investigate the expression profiles and biofunctions of CLCs genes, and construct a novel risk signature to predict biochemical recurrence (BCR) of PCa patients. We identified five differentially expressed CLCs genes in our cohort and then constructed a signature composed of CLCN2 and CLCN6 through Lasso-Cox regression analysis in the training cohort from the Cancer Genome Atlas (TCGA). The testing and entire cohorts from TCGA and the GSE21034 from the Gene Expression Omnibus (GEO) were used as internal and independent external validation datasets. This signature could divide PCa patients into the high and low risk groups with different prognoses, was apparently correlated with clinical features, and was an independent excellent prognostic indicator. Enrichment analysis indicated our signature was primarily concentrated in cellular process and metabolic process. The expression patterns of CLCN2 and CLCN6 were detected in our own cohort based immunohistochemistry staining, and we found CLCN2 and CLCN6 were highly expressed in PCa tissues compared with benign tissues and positively associated with higher Gleason score and shorter BCR-free time. Functional experiments revealed that CLCN2 and CLCN6 downregulation inhibited cell proliferation, colony formation, invasion, and migration, but prolonged cell cycle and promoted apoptosis. Furthermore, Seahorse assay showed that silencing CLCN2 or CLCN6 exerted potential inhibitory effects on energy metabolism in PCa. Collectively, our signature could provide a novel and robust strategy for the prognostic evaluation and improve treatment decision making for PCa patients.

The aim of this study is to construct an inflammatory response-related genes (IRRGs) signature to monitor biochemical recurrence (BCR) and treatment effects in prostate cancer patients (PCa). A gene signature for inflammatory responses was constructed on the basis of the data from the Cancer Genome Atlas (TCGA) database, and validated in external datasets. It was analyzed using receiver operating characteristic curve, BCR-free survival, Cox regression, and nomogram. Distribution analysis and external model comparison were utilized. Then, enrichment analysis, tumor mutation burden, tumor immune microenvironment, and immune cell infiltration signatures were investigated. The role of the signature in immunotherapy was evaluated. The expression patterns of core genes were verified by RNA sequencing. We identified an IRRGs signature in the TCGA-PRAD cohort and verified it well in two other independent external datasets. The signature was a robust and independent prognostic index for predicting the BCR of PCa. The high-risk group of our signature predicted a shortened BCR time and an aggressive disease progression. A nomogram was constructed to predict BCR-free time in clinical practices. Neutrophils and CD8+ T cells were in higher abundance among the low-risk individuals. Immune functions varied significantly between the two groups and immune checkpoint therapy worked better for the low-risk patients. The expression of four IRRGs showed significant differences between PCa and surrounding benign tissues, and were validated in BPH-1 and DU145 cell lines by RNA sequencing. Our signature served as a reliable and promising biomarker for predicting the prognosis and evaluating the efficacy of immunotherapy, facilitating a better outcome for PCa patients.

Objective:To investigate the experience, safety and feasibility of robot assisted simple prostatectomy for large volume benign prostatic hyperplasia (BPH).Methods:The clinical data of 21 patients who underwent robot assisted simple prostatectomy in our hospital from Jan. 2017 to Jan. 2021 were retrospectively analyzed, and the surgical procedures, intraoperative and postoperative problems were summarized.Results:All cases were operated successfully without conversion to open surgery. The average operation time was (135.7±23.2) min. The average blood loss was (168.4±21.5) ml without blood transfusion in all cases. Bladder irrigation time was within 24 h after operation. The average suprapubic catheter time was 2 days. The average drain time was 3 days, and the mean catheter time was (12.5±2.4) days. Post-void residual, international prostate symptom score (IPSS) and maximum urine flow were improved significantly 3 months after operation (P<0.01).Conclusions:Transperitoneal robot assisted simple prostatectomy is a safe and effective minimally invasive method for large volume benign prostatic hyperplasia, which may become an alternative treatment.