1.The effect of DNA methyltransferase 3b (DNMT3b) stable knockdown on Adriamycin resistance of bladder cancer cell
Ke CHEN ; Jing XIE ; Yingsheng LIN ; Kefeng XIAO ; Jinan GUO
Chinese Journal of Urology 2025;46(6):455-461
Objective:To investigate the effect of stable low expression of DNA methyltransferase 3b(DNMT3b)on the adriamycin(ADM)resistance of wild-type bladder cancer cells BIU-87 and BIU-87/ADM-resistant cells(BIU-87/ADM).Methods:Lentiviruses expressing DNMT3b siRNA and negative control siRNA were packaged. Stable DNMT3b-low-expressing BIU-87 cells(BIU-87-siRNA group),BIU-87/ADM cells(BIU-87/ADM-siRNA group),and corresponding control groups(BIU-87-NC group and BIU-87/ADM-NC group)were established via lentiviral infection. DNMT3b expression was detected by quantitative PCR and Western blot to validate siRNA interference efficiency. BIU-87-siRNA and BIU-87-NC cells were treated with 0.0125,0.025,0.05,0.1,and 0.2 mg/L ADM,while BIU-87/ADM-siRNA and BIU-87/ADM-NC cells were treated with 0.5,1,2,4,and 8 mg/L ADM. Cell survival rates were measured using the MTT assay to calculate the half-maximal inhibitory concentration(IC50)and relative reversal rate. Apoptosis was analyzed by flow cytometry. Expression of drug resistance-related genes(MRP1,P-gp and Survivin)was detected by quantitative PCR and Western blot. In vivo tumorigenesis experiments were performed by subcutaneously inoculating BIU-87-siRNA,BIU-87/ADM-siRNA,and control group cells into nude mice. Tumor sizes were measured on days 7,10,13,16,20,and 25 to plot growth curves and assess the effect of DNMT3b low expression on ADM resistance.Results:Quantitative PCR showed that the relative mRNA expression of DNMT3b in BIU-87-siRNA and BIU-87-NC groups were(0.32 ± 0.08) vs.(1.00±0.12)( P < 0.01),and(0.30 ± 0.07) vs.(1.00 ± 0.11)in BIU-87/ADM-siRNA vs. BIU-87/ADM-NC groups( P < 0.01). Western blot confirmed significantly reduced DNMT3b protein levels in both siRNA groups( P < 0.01). After ADM treatment,BIU-87-siRNA cells exhibited lower survival rates compared to BIU-87-NC at 0.025,0.05,and 0.1 mg/L ADM( P < 0.05),with IC50 values of(0.14 ± 0.02)mg/L vs.(0.18 ± 0.03)mg/L( P > 0.05). For BIU-87/ADM-siRNA cells,survival rates at 1,2,4,and 8 mg/L ADM were significantly lower than controls( P < 0.05),with IC50 values of(7.10 ± 0.45)mg/L vs.(13.96 ± 1.20)mg/L and a relative reversal rate of 49.76%( P < 0.01). Apoptosis rates were significantly higher in siRNA groups( P < 0.01). mRNA and protein levels of MRP1,P-gp,and Survivin were reduced in both siRNA groups( P < 0.05),except for P-gp protein in BIU-87-siRNA cells( P > 0.05). In vivo,tumor volumes in siRNA groups were significantly smaller than controls by day 25( P < 0.05). Conclusion:Stable low expression of DNMT3b reverses ADM resistance in bladder cancer BIU-87/ADM cells.
2.The effect of DNA methyltransferase 3b (DNMT3b) stable knockdown on Adriamycin resistance of bladder cancer cell
Ke CHEN ; Jing XIE ; Yingsheng LIN ; Kefeng XIAO ; Jinan GUO
Chinese Journal of Urology 2025;46(6):455-461
Objective:To investigate the effect of stable low expression of DNA methyltransferase 3b(DNMT3b)on the adriamycin(ADM)resistance of wild-type bladder cancer cells BIU-87 and BIU-87/ADM-resistant cells(BIU-87/ADM).Methods:Lentiviruses expressing DNMT3b siRNA and negative control siRNA were packaged. Stable DNMT3b-low-expressing BIU-87 cells(BIU-87-siRNA group),BIU-87/ADM cells(BIU-87/ADM-siRNA group),and corresponding control groups(BIU-87-NC group and BIU-87/ADM-NC group)were established via lentiviral infection. DNMT3b expression was detected by quantitative PCR and Western blot to validate siRNA interference efficiency. BIU-87-siRNA and BIU-87-NC cells were treated with 0.0125,0.025,0.05,0.1,and 0.2 mg/L ADM,while BIU-87/ADM-siRNA and BIU-87/ADM-NC cells were treated with 0.5,1,2,4,and 8 mg/L ADM. Cell survival rates were measured using the MTT assay to calculate the half-maximal inhibitory concentration(IC50)and relative reversal rate. Apoptosis was analyzed by flow cytometry. Expression of drug resistance-related genes(MRP1,P-gp and Survivin)was detected by quantitative PCR and Western blot. In vivo tumorigenesis experiments were performed by subcutaneously inoculating BIU-87-siRNA,BIU-87/ADM-siRNA,and control group cells into nude mice. Tumor sizes were measured on days 7,10,13,16,20,and 25 to plot growth curves and assess the effect of DNMT3b low expression on ADM resistance.Results:Quantitative PCR showed that the relative mRNA expression of DNMT3b in BIU-87-siRNA and BIU-87-NC groups were(0.32 ± 0.08) vs.(1.00±0.12)( P < 0.01),and(0.30 ± 0.07) vs.(1.00 ± 0.11)in BIU-87/ADM-siRNA vs. BIU-87/ADM-NC groups( P < 0.01). Western blot confirmed significantly reduced DNMT3b protein levels in both siRNA groups( P < 0.01). After ADM treatment,BIU-87-siRNA cells exhibited lower survival rates compared to BIU-87-NC at 0.025,0.05,and 0.1 mg/L ADM( P < 0.05),with IC50 values of(0.14 ± 0.02)mg/L vs.(0.18 ± 0.03)mg/L( P > 0.05). For BIU-87/ADM-siRNA cells,survival rates at 1,2,4,and 8 mg/L ADM were significantly lower than controls( P < 0.05),with IC50 values of(7.10 ± 0.45)mg/L vs.(13.96 ± 1.20)mg/L and a relative reversal rate of 49.76%( P < 0.01). Apoptosis rates were significantly higher in siRNA groups( P < 0.01). mRNA and protein levels of MRP1,P-gp,and Survivin were reduced in both siRNA groups( P < 0.05),except for P-gp protein in BIU-87-siRNA cells( P > 0.05). In vivo,tumor volumes in siRNA groups were significantly smaller than controls by day 25( P < 0.05). Conclusion:Stable low expression of DNMT3b reverses ADM resistance in bladder cancer BIU-87/ADM cells.
3.Advances in active surveillance of prostate cancer
Yingsheng LIN ; Zengqin LIU ; Jing XIE ; Kefeng XIAO
Chinese Journal of Urology 2023;44(9):717-720
The popularity of prostate-specific antigen screening has led to overdiagnosis and overtreatment of prostate cancer. Active surveillance can be used as a safe and reliable treatment option for some patients with localized prostate cancer. At the same time, compared with surgery and chemoradiotherapy, active surveillance can provide these patients with a better quality of life. A variety of new monitoring methods, including PSMA PET and biomarkers, can optimize the risk stratification of prostate cancer and reduce unnecessary puncture biopsies to further improve the quality of life of patients. However, there are still great controversies about the inclusion criteria, monitoring programs, and intervention indications for active surveillance of prostate cancer, and domestic doctors and patients still lack acceptance of it. This article reviews the research progress of active surveillance in prostate cancer.
4.Comparison of efficacy and safety between biphasic insulin aspart 50 and biphasic human insulin 50:A randomized crossover trial
Xiaohui GUO ; Fang BIAN ; Yumei DONG ; Hong TANG ; Jian TIAN ; Guixia WANG ; Tao YANG ; Yufeng LI ; Yingsheng ZHOU ; Dalong ZHU ; Shan HUANG ; Jing LIN ; Shi ZHAO ; Jian WANG ; Lei GE ; Yi QU ; Yan GAO
Chinese Journal of Endocrinology and Metabolism 2016;32(7):564-571
Objective To investigate the efficacy of biphasic insulin aspart 50(BIAsp50)twice daily(bid) versusbiphasichumaninsulin50(BHI50)(bid)plusmetforminonbloodglucosecontrolfollowingastandardmealtest in Chinese patients with type 2 diabetes mellitus(T2DM). Methods A randomized, open-label, 2-sequence, crossover trial for two 4-week treatment periods was conducted in 14 Chines institutes. Eligible subjects inadequately controlled with BHI50(bid)plus metformin were randomized to two sequences in a 1 : 1 ratio(A:BIAsp50-BHI50, B:BHI50-BIAsp50 ) . Standard meal tests were performed at baseline and the ends of two periods within 4 weeks. Primary endpoint was 2h postprandial plasma glucose ( PPG) increment following standard meal test, with insulin dose standardized at 0. 3 IU/kg. Results A total of 161 subjects were randomized into two sequences(81 to sequence A, and 80 to sequence B) and finally analysed. After 4 weeks of treatment, mean 2h PPG increment with BIAsp50 was lower than that with BHI50 [ treatment difference of BIAsp50 vs BHI50: -1. 12 mmol/L ( 95% CI-1. 66,-0. 58), P<0. 01], suggesting superiority of BIAsp50 over BHI50. Incremental area under the curve for PPG(0-2 h)with BIAsp50 was lower than that with BHI50 [treatment difference:-38. 8 mmol·L-1·min-1(95%CI-77. 3,-0. 26), P=0. 049], as was the mean 2h PPG [treatment difference:-0. 58 mmol/L(95% CI -1. 13,-0. 03), P=0. 040]. The FPG value with BIAsp50 was higher than that with BHI50 [treatment difference:0. 52 mmol/L(95%CI 0. 18, 0. 86), P=0. 003]. The rate of nocturnal hypoglycemia with BIAsp50 was lower than that with BHI50(1. 13 vs 2. 86 events per subject year, P<0. 01). Conclusion In patients with T2DM inadequately controlled with BHI50 plus metformin, BIAsp50 was proven to be well-tolerated with improved postprandial glucose control compared with BHI50.
5.Relative factors and treatment of the ununited fractures of the diaphysis of radius and ulna
Tingjun LIN ; Shenglian XU ; Yingsheng DENG ; Chongwen ZOU ; Xueyong YANG ;
Chinese Journal of Orthopaedic Trauma 2002;0(03):-
Objective To explore the possible causes and treatment of nonunion of fractures of diaphysis of radius and ulna. Methods According to different personal conditions and results of X gram, 73 cases of the nonunion fractures of diaphysis of radius and ulna were treated with different internal fixations, autologus spongy bone transplantation, release of scar and interosseous membrane, or local skin flap transposition. All had early functional mobilization of the forearm after operation. Results The patients were followed up for 12 to 29 months. All the cases had bony union, and 64 cases(87.7% ) obtained satisfactory functional rehabilitation. Conclusion The major causes that may lead to the nonunion of the diaphysis of radius and ulna are conditions of local soft tissue, site and severity of the fracture, security of internal fixation, and improper early movement, rather than infection. Proper internal fixation, autologus spongy bone transplantation, and release of scar and interosseous membrane are effective in treatment of the nonunion of the fracture and in the functional rehabilitation of the forearms.

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