1.Role of N6-methyladenosine methylation related gene IGF2BP3 in renal clear cell carcinoma
Xianzhong WANG ; Zhong WANG ; Dapeng HE ; Yaming LAI ; Wenya AN ; Zugang SUN ; Xisong XIE ; Chunxiang WU
International Journal of Laboratory Medicine 2025;46(1):27-31
Objective To screen the key N6-methyladenosine(m6A)methylation related genes in renal clear cell carcinoma(ccRCC),and to study their expression and relationship with the prognosis,migration and invasion of renal clear cell carcinoma.Methods The RNA sequencing data and clinical data of ccRCC and ad-jacent tissues were downloaded from the Cancer Genome Atlas(TCGA)and GTEx(Genotype-Tissue Expres-sion).The expression profile and prognosis were analyzed with R 4.1.1,and the key genes were screened.Clinical specimens of 10 patients with ccRCC were collected.The mRNA and protein expressions were detec-ted by RT-qPCR and immunohistochemistry,respectively.In human ccRCC cell line RCC23,siRNA was used to knock down key genes,and CCK-8 was used to detect the survival rate of cells.Scratch test and Trans well test were used to detect the migration and invasion of cells,respectively.Results Among the 19 m6A methyl-ation related genes,only insulin-like growth factor 2 mRNA binding protein 3(IGF2BP3)was highly ex-pressed in cancer tissues,and the high expression was significantly positively correlated with poor prognosis.The high expression of IGF2BP3 was verified in clinical specimens by RT-qPCR and immunohistochemistry.After knockdown of IGF2BP3 by siRNA,the survival rate of RCC23 cells decreased significantly,and the mi-gration and invasion ability of cut cells decreased.Conclusion These results suggest that IGF2BP3 may be an effective biomarker and potential drug target for predicting the prognosis of patients with ccRCC.
2.Naringenin regulates RAW264.7 macrophage polarization induced by high glucose through RhoA/ROCK signaling pathway
Liping YOU ; Fajiang XIE ; Jian FENG ; Zhuo LAN ; Xisong HE ; Yafei LI ; Jiafu LI
Journal of Xi'an Jiaotong University(Medical Sciences) 2021;42(2):225-232
【Objective】 To investigate the effects of naringenin on the polarization of high-glycemic RAW264.7 macrophages and its related mechanism. 【Methods】 The mother solution of NAR was prepared with dimethyl sulfoxide (DMSO), and the log-growth phase macrophage RAW264.7 was pre-tested. DMEM medium with different glucose concentrations (1, 2, 4, 5 and 6 g/L) was used for cultivation for 24 h. Before the experiment, DMEM was diluted into NAR mixture with different final concentrations, and the effect of NAR on RAW264.7 cell activity was detected by CCK-8 method; nitric oxide synthase (NOS) type classification determination of checkerboard induced nitric oxide synthase (iNOS) active filter was used to control the concentration of sugar and high-sugar stimulation. The control group were subdivided into normal control (NG) and osmotic pressure control (NG+M). The high-glucose stimulation group was divided into normal high glucose (HG), high glucose + naringenin (HG+NAR), high glucose + Fasudil (HG+F), and high glucose +C3 transferase (HG+C3). RAW264.7 was cultured for 24 h in each group; the expression levels of supernatant cytokines, namely, interleukin6 (il-6), tumor necrosis factor -α (TNF-α) and interleukin10 (IL-10), were detected by ELISA. Western blotting was used to determine the RhoA/ROCK pathway related proteins, iNOS and Arg-1 protein levels. Type (M1, M2) and proportion (M1/M2) of macrophages were analyzed by flow cytometry. 【Results】 Compared with those in NG group, in HG group RhoA/ROCK pathway-related proteins and iNOS expression were increased, while Arg-1 expression was decreased (P<0.05). The secretion of pro-inflammatory cytokines IL-6 and TNF-α was increased while anti-inflammatory cytokine IL-10 was decreased (P<0.05). The number of M1-type cells and M1/M2 ratio increased (P<0.05). Compared with HG group, RhoA/ROCK pathway related proteins and iNOS expression were decreased in HG+NAR group, HG+F group and HG+C3 group, while Arg-1 expression was increased, IL-6 and TNF- α secretion was decreased, IL-10 was increased, M2-type macrophages were increased, and M1/M2 was decreased (P<0.05). 【Conclusion】 NAR may promote the M2-type differentiation of macrophages stimulated by high glucose by down-regulating RhoA/ROCK signaling pathway.
3.Effect of postoperative stent placement time on the outcome of flexible ureteroscope lithotripsy in the treatment of kidney calculi
Xianzhong WANG ; Zhong WANG ; Dapeng HE ; Xisong XIE
International Journal of Surgery 2020;47(9):619-623
Objective:To explore the effect of postoperative stent placement time on flexible ureteroscopy lithotripsy (FURL) in the treatment of kidney calculi.Methods:A retrospective analysis of 126 patients with kidney calculi admitted to Guangyuan Central Hospital from April 2017 to November 2018. All patients were treated with FURL. The patients were divided into two groups according to the time of stent placement after FURL: conventional tube placement group ( n=91) and short-term tube placement group ( n=35). The postoperative stent placement time of patients in the conventional tube placement group was 4 weeks, and the postoperative stent placement time of patients in the short-term tube placement group was 2 weeks. The postoperative outcome indicators (hospitalization time and stone-free rate), postoperative complication indicators (postoperative hematuria, renal hematoma, urinary tract infection and fever) and postoperative pain were compared between the two groups. The measurement data was expressed as mean±standard deviation ( Mean± SD), and the Student′s t test was used for comparison between groups; the Chi-square test was used for comparison between counting data. Results:There were not significant differences in hospitalization time [(7.22±1.02) d vs (7.26±1.22) d], stone-free rate 1 month after surgery (84.6% vs 82.8%), stone-free rate 2 month after surgery (93.4% vs 88.5%), postoperative hematuria (10.9% vs 11.4%), renal hematoma (3.3% vs 0) and fever (2.2% vs 0) between the conventional tube placement group and short-term tube placement group ( P>0.05). Compared with the conventional tube placement group, the short-term tube group had lower urinary tract infection rates (17.6% vs 2.9%), the difference was statistically significant ( P<0.05). There was not significant difference in the average use rate of opioids [(1.05±0.45) vs (1.00±0.50), P>0.05], acetaminophen and nonsteroidal anti-inflammatory drugs (NSAIDs) [(1.52±0.55) vs (1.45±0.45), P>0.05] between the two groups during hospitalization. There was not statistically significant difference in the incidence of abdominal pain after discharge from the hospital [(1.25±0.55) vs (1.15±0.45)]. The number of applications of acetaminophen and NSAIDs in the conventional tube placement group was greater than that in the short-term tube placement group [(1.45±0.46) vs (1.25±0.30)] after one month, the difference was statistically significant ( P<0.05). Conclusions:Short-term tube placement after renal FURL surgery is as effective as traditional long-term stenting, achieving the same stone-free effect and reducing early postoperative complications. For the reduction of postoperative infection and postoperative pain, short-term stenting has certain advantages.

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