Objective To investigate the expression characteristics of the transcription factor E2F2 in clear cell renal cell carcinoma (ccRCC), its impact on patient prognosis, and its potential role in the tumor immune microenvironment, with validation using clinical samples and functional assays. Methods RNA sequencing data and clinical information of ccRCC patients were obtained from the TCGA database; GSE53757 and GSE66272 datasets were downloaded from the GEO database as external validation cohorts. The expression difference of E2F2 between ccRCC tissue and normal tissue was compared, and the relationship between E2F2 expression and clinical pathological characteristics was analyzed. Kaplan-Meier (KM) survival analysis with log-rank test was performed to evaluate the effects of E2F2 on overall survival (OS), progression-free survival (PFS), and disease-specific survival (DSS). The prognostic efficacy of E2F2 in predicting 1-, 3- and 5-year survival was evaluated using receiver operating characteristic (ROC) curve. GSEA was performed to identify E2F2-related signaling pathways, and the ESTIMATE and CIBERSORT algorithms were used to assess the relationship between E2F2 expression level and tumor immune cell infiltration and immune microenvironment. Twenty pairs of surgically resected and pathologically confirmed ccRCC tissues and matched adjacent non-tumor tissues were collected from our hospital, and immunohistochemistry (IHC) was performed to detect the protein expression of E2F2. Human ccRCC cell line 786-O was used for functional assays;shRNA was used to knock down E2F2 expression (constructing stable shE2F2#1 and shE2F2#2 cell lines), and qRT-PCR and Western blot were performed to verify knockdown efficiency, followed by MTT, colony formation, and Transwell migration assays to evaluate the effects of E2F2 on the biological behaviors of ccRCC cells. Results E2F2 was significantly upregulated in ccRCC tissues (the same results in the verification set). Higher E2F2 expression was associated with advanced histological grade, clinical stage and higher TNM classification (P＜0.05). KM analysis showed that patients in the E2F2 high-expression group had worse OS, PFS, and DSS (P=0.019, 0.036, ＜0.001); the ROC curves showed area under the curve (AUC) of E2F2 of predicting 1-, 3- and 5- year survival of ccRCC patients were 0.844, 0.851 and 0.815, respectively. GSEA revealed that the E2F2 low-expression group was enriched in multiple metabolism-related pathways, whereas the E2F2 high-expression group was associated with immune-related pathways. Immune analysis demonstrated that high E2F2 expression was associated with increased immune scores, increased proportion of immune cells, higher tumor mutation burden, and potentially stronger immunotherapy response. IHC results showed that the E2F2 immunoreactivity score in ccRCC tissues was significantly higher than that in adjacent non-tumor tissues (P＜0.05). Functional assays indicated that E2F2 knockdown significantly inhibited the proliferation, colony formation, and migration of ccRCC cells. Conclusion E2F2 is highly expressed in ccRCC and may be involved in tumorigenesis and progression through modulation of the immune microenvironment. Its expression level is closely associated with patient prognosis, and E2F2 has the potential to serve as a prognostic biomarker and immunotherapeutic target in ccRCC.

Objective To investigate the value of heparin-binding protein(HBP) in predicting urosepsis after endoscopic lithotripsy for upper urinary tract stones.Methods From January to November 2018,635 cases of upper urinary tract stones treated by endoscopic lithotripsy in our hospital was analyzed.According to the 2014 edition Chinese Guidelines of the diagnosis and treatment of urosepsis,all patients were divided into two groups,the definite urosepsis group (n =53,8.3％) and the no urosepsis group (n =582,91.7％).The serum of HBP level was detected by fluorescence immunoassay technology.The serum of procalcitonin (PCT) level was detected by cyclic enhanced immunofluorescent assay.C-reactive protein (CRP) level was detected by automatic chemiluminescence immunoassay analyzer.White blood cells (WBC) count was performed by hematology analyzer.Receiver operating characteristic(ROC) curves were constructed by SPSS 22 and MedCale,and areas under curves (AUC) were calculated and compared to illustrate the diagnostic power of HBP,PCT,CRP and WBC.The correlation between HBP and WBC was also significant (r =0.68,P ＜ 0.05).Results The level of HBP,PCT,CRP and WBC were significantly higher in the definite urosepsis group compared to the no urosepsis group.AUC of HBP,PCT,CRP and WBC were 0.930,0.878,0.727 and 0.617.When the cutoff of HBP,PCT,CRP and WBC were set as 51.5 ng/ml,0.41 ng/ml,64.2 mg/L and 11.7 × 109/L,the sensitivities were 89.2％,83.3％,83.5％ and 44.7％,and the specificity were 95.9％,75.6％,64.3％ and 77.1％,respectively.Conclusions The value of heparin-binding protein was superior to that of PCT,CRP and WBC in predicting urosepsis after endoscopic lithotripsy for upper urinary tract stones.Heparin-binding protein could be acted as a diagnostic marker for urosepsis.