[Objective]This study aims to investigate the effects of methionine sulfoxide reductase A(MSRA)on the proliferation,apoptosis,invasion,and metastasis of clear cell renal cell carcinoma(ccRCC)through cellular and animal experiments to elucidate its potential biological mechanisms,providing new molecular targets and strategies for the treatment of ccRCC.[Methods]MSRA-overexpressing cell lines were constructed with transfection.The effects of MSRA on the proliferation,apoptosis,invasion,and migration of ccRCC cells were assessed through proliferation assays,colony formation assays,apoptosis assays,wound healing assays,and invasion assays.Further,the mechanisms and related signaling pathways were explored via ROS detection,RT-qPCR,and Western blot.Subsequently,a subcutaneous xenograft mouse model was employed to verify the effect of MSRA on ccRCC tumor growth and metastasis at the animal level to explore its potential molecular mechanisms.[Results]The mRNA and protein expressions of MSRA in OS-RC-2 and 786-O cell lines were low(RT-qPCR:0.57±0.09,0.56±0.04;WB:0.26±0.13,0.24±0.09).RT-qPCR and Western blot experiments confirmed the successful construction of OS-RC-2-pLVSO2-MSRA(RT-qPCR:108.04±1.80;WB:117.01±20.19)and 786-O-pLVSO2-MSRA(973.45±51.37;WB:190.34±30.13)overexpressed cell lines,with statistically significant differences(P＜0.001).Proliferation assays showed reduced proliferation in OS-RC-2-pLVSO2-MSRA(72 h:1.246±0.003)and 786-O-pLVSO2-MSRA(72 h:1.468±0.001),with significant differences(P＜0.001).Colony formation assays revealed a decrease in colony numbers in OS-RC-2-pLVSO2-MSRA(0.090±0.002)and 786-O-pLVSO2-MSRA(0.080±0.002),with significant differences(P＜0.001).Apoptosis assays demonstrated increased apoptosis rates in OS-RC-2-pLVSO2-MSRA(2.013±0.116)and 786-O-pLVSO2-MSRA(4.767±0.199),with significant differences(P＜0.001).Wound healing assays indicated less migration distance in OS-RC-2-pLVSO2-MSRA(0.643±0.028)and 786-O-pLVSO2-MSRA(0.603±0.034),with significant differences(P＜0.001).Transwell assays showed a reduction in the numbers of penetrative cells in OS-RC-2-pLVSO2-MSRA(16.80±2.28)and 786-O-pLVSO2-MSRA(21.40±4.78),with significant differences(P＜0.001).Fluorescence assays indicated a decreased ROS levels in OS-RC-2-pLVSO2-MSRA(50.59±6.24)and 786-O-pLVSO2-MSRA(62.87±5.35),with significant differences(P＜0.001).Western blot analysis showed a decrease in the expression of N-cadherin and Vimentin,and an increase in the expression of E-cadherin in OS-RC-2-pLVSO2-MSRA and 786-O-pLVSO2-MSRA(P＜0.001).Western blot analysis revealed a significant decrease in the expression levels of p-ERK1/2 and p-SMAD3 in OS-RC-2-pLVSO2-MSRA and 786-O-pLVSO2-MSRA(P＜0.001).Animal experiments showed reduced tumor volumes in OS-RC-2-pLVSO2-MSRA(155.00±50.46),with significant differences(P＜0.001).Western blot analysis of tumor tissues from animals confirmed the decreased expression of N-cadherin and Vimentin,and the increased expression of E-cadherin,with significant differences(P＜0.001).IHC experiments of OS-RC-2-pLVSO2-MSRA tumors revealed a decrease in the expression of Ki-67,N-cadherin,and Vimentin,and an increase in the expression of E-cadherin,with significant differences(P＜0.001).[Conclusion]MSRA overexpression inhibits ROS expression in ccRCC,suppresses the EMT process,and consequently inhibits the proliferation,invasion,and metastasis of ccRCC.

[Objective]This study aims to investigate the effects of methionine sulfoxide reductase A(MSRA)on the proliferation,apoptosis,invasion,and metastasis of clear cell renal cell carcinoma(ccRCC)through cellular and animal experiments to elucidate its potential biological mechanisms,providing new molecular targets and strategies for the treatment of ccRCC.[Methods]MSRA-overexpressing cell lines were constructed with transfection.The effects of MSRA on the proliferation,apoptosis,invasion,and migration of ccRCC cells were assessed through proliferation assays,colony formation assays,apoptosis assays,wound healing assays,and invasion assays.Further,the mechanisms and related signaling pathways were explored via ROS detection,RT-qPCR,and Western blot.Subsequently,a subcutaneous xenograft mouse model was employed to verify the effect of MSRA on ccRCC tumor growth and metastasis at the animal level to explore its potential molecular mechanisms.[Results]The mRNA and protein expressions of MSRA in OS-RC-2 and 786-O cell lines were low(RT-qPCR:0.57±0.09,0.56±0.04;WB:0.26±0.13,0.24±0.09).RT-qPCR and Western blot experiments confirmed the successful construction of OS-RC-2-pLVSO2-MSRA(RT-qPCR:108.04±1.80;WB:117.01±20.19)and 786-O-pLVSO2-MSRA(973.45±51.37;WB:190.34±30.13)overexpressed cell lines,with statistically significant differences(P＜0.001).Proliferation assays showed reduced proliferation in OS-RC-2-pLVSO2-MSRA(72 h:1.246±0.003)and 786-O-pLVSO2-MSRA(72 h:1.468±0.001),with significant differences(P＜0.001).Colony formation assays revealed a decrease in colony numbers in OS-RC-2-pLVSO2-MSRA(0.090±0.002)and 786-O-pLVSO2-MSRA(0.080±0.002),with significant differences(P＜0.001).Apoptosis assays demonstrated increased apoptosis rates in OS-RC-2-pLVSO2-MSRA(2.013±0.116)and 786-O-pLVSO2-MSRA(4.767±0.199),with significant differences(P＜0.001).Wound healing assays indicated less migration distance in OS-RC-2-pLVSO2-MSRA(0.643±0.028)and 786-O-pLVSO2-MSRA(0.603±0.034),with significant differences(P＜0.001).Transwell assays showed a reduction in the numbers of penetrative cells in OS-RC-2-pLVSO2-MSRA(16.80±2.28)and 786-O-pLVSO2-MSRA(21.40±4.78),with significant differences(P＜0.001).Fluorescence assays indicated a decreased ROS levels in OS-RC-2-pLVSO2-MSRA(50.59±6.24)and 786-O-pLVSO2-MSRA(62.87±5.35),with significant differences(P＜0.001).Western blot analysis showed a decrease in the expression of N-cadherin and Vimentin,and an increase in the expression of E-cadherin in OS-RC-2-pLVSO2-MSRA and 786-O-pLVSO2-MSRA(P＜0.001).Western blot analysis revealed a significant decrease in the expression levels of p-ERK1/2 and p-SMAD3 in OS-RC-2-pLVSO2-MSRA and 786-O-pLVSO2-MSRA(P＜0.001).Animal experiments showed reduced tumor volumes in OS-RC-2-pLVSO2-MSRA(155.00±50.46),with significant differences(P＜0.001).Western blot analysis of tumor tissues from animals confirmed the decreased expression of N-cadherin and Vimentin,and the increased expression of E-cadherin,with significant differences(P＜0.001).IHC experiments of OS-RC-2-pLVSO2-MSRA tumors revealed a decrease in the expression of Ki-67,N-cadherin,and Vimentin,and an increase in the expression of E-cadherin,with significant differences(P＜0.001).[Conclusion]MSRA overexpression inhibits ROS expression in ccRCC,suppresses the EMT process,and consequently inhibits the proliferation,invasion,and metastasis of ccRCC.

Objective To detecte the expression of Apelin ( APLN) in prostate cancer tissue and to investi-gate its correlation with prognosis of prostate cancer .Methods Collect prostate cancer patients in Department of Urology of Huadu District People's Hospital for 2014 -2016 years in Guangzhou .The expression of APLN was detected by real-time fluorescence quantitative polymerase chain reaction ( qRT-PCR) in 20 prostate cancer tissues and 20 adjacent normal tissues.The difference between the two groups was compared .And 104 samples of primary prostate cancer and 28 samples of benign paracancerous tissues from the Taylor database were selected to analyze its relationship with the clinical features and prognosis of the patients .Results Compared with the benign paracancerous tissue(7.26 ±0.03),APLN was up-regulated in the prostate cancer tissue (7.62 ±0.42)(t=3.824,P<0.001). The up-regulation of APLN was associated with pathological stage (t=2.942,P=0.003),metastasis(t=3.022, P<0.001),Gleason score (t =2.399,P =0.031),the biochemical recurrence -free survival(t =2.533,P =0.001 ) ,and the biochemical recurrence -free survival of the patients with higher expression of APLN was shorter than that of the patients with lower expression of APLN(χ2 =6.268,P=0.012).Conclusion The abnormal expres-sion of APLN may be associated with tumor formation and malignant progression of prostate cancer .High expression of APLN can predict the biochemical recurrence -free survival in patients with prostate cancer .

Objective To evaluate the efficacy and safety of intradetrusor injection of botulinum toxin type A in the treatment of ketamine-related cystitis.Methods A retrospective analysis of clinical data of 36 ketamine-related cystitis patients with intradetrusor injection of botulinum toxin type A treatment in our hospital during August 2010 to December 2015 was conducted, including 31 males and 5 females with a mean age of 25.5 years.All patients had failed to conventional treatment options including cessation of ketamine, antibiotics, M-blockers.At the time of the first injection, patients were injected with 200 U botulinum toxin type A diluted in 15 ml of 0.9％ saline into the detrusor muscle at 30 sites, sparing the trigone, under cystoscopic guidance.3-d voiding diary, interstitial cystitis symptom index (ICSI),interstitial cystitis problem index (ICPI), pelvic pain and urinary frequency/urgency symptom score (PUF) were recorded to evaluate the efficacy.The treatment-related complications were recorded.When the efficacy of botulinum toxin type A decreased and the patient's symptoms returned to baseline before treatment, the patient received repeated injections of botulinum toxin type A with the same dose and method as the first injection.Results Thirty-six patients with ketamine-related cystitis were treated with intradetrusor injection of botulinum toxin type A.Sixteen patients received two injection treatments and two patients received three injection treatments.During the follow-up, 3-d urinary diaries, ICSI, ICPI, and PUF showed a significant improvement in outcome at 4 weeks after the first injection.The efficacy of the second and third injection treatment was also remarkable.Three patients developed urinary tract infection after the first injection, and two patients developed urinary tract infection after the second injection.Mild hematuria occurred in 15 patients after the first injection, and mild hematuria occurred in 7 patients after the second injection, which was improved in 1 to 2 days.All patients did not appear acute urinary retention and other adverse drug reactions.Conclusions Intradetrusor injection of botulinum toxin type A could be a safe and effective method for the treatment of ketamine-related cystitis.Repeated injection therapy is still safe and effective.

Objective We used the dataset base on high throughput sequencing data to construct a diagnosis model by ANN and GA.Methods We screened the Taylor_prostate datasets from GEO according to,then we used the GA to screen the datas further. Finally we used the ANN to analyze the datas and construct a diagnosis model. To validate the model,we used 10-folds crossvalidation as the inner validation and the datas from Grasso dataset( GPL6480 and GPL6848) were used as the outter validation.Results We got 5 genes ACADL,ACTG2, CACNA2D1,PCP4 and SPARCL1.And we used spss to get the AUC of the model which is 94.62.The result of validation is good.Conclusion The performance of the model is good because the AUC is larger than 0.5.

Objective To study the effects of IL-4 in prostate cancer cells glycometabolism and proliferation. Methods We used IL-4 to treat PC3 cells, then tested the changes of LDH-A expression by RT-qPCR, Western Blot, CCK-8 and lactate production assay. Results Our data showed that IL-4 induced LDH-A up-expression in PC3 cells at mRNA and protein levels. Also, IL-4 promoted the proliferation activation and increased lactate production in PC3 cells. Conclusion IL-4 can strengthen the proliferation activation in PC3 cells by up-regulating LDH-A expression.

Objective To investigate the safety and efficacy of nephron-sparing surgery (NSS)for selective T2 stage renal tumor.Methods The surgical database of 26 patients treated with NSS for clinical T2 stage renal cell carcinomas between March 2010 and May 2013 were collected and analyzed retrospectively.There were 17 males and 9 females,with a mean age of 52 years (39-74 years),mean tumor size of 10.3 cm(7.2-16.5 cm),and mean R.E.N.A.L score of 7.5 (6-10).Patients'demographics,clinical characteristics,oncologic outcomes,renal function were reviewed.Results The renal masses were removed successfully and the surgical margins were negative.There were 21 (80.8％) cases of clear cell carcinoma,4 (15.4％) papillary carcinoma and 1 (3.8％) chromophobe carcinoma.The mean ischemia time was (28.3 ± 12.5) minutes (7 patients were clamp-free).Three patients needed transfusion,one experienced urine fistula and cured by conservative treatment,and one patient's renal function got progressive worsening and required long-term hemodialysis.The average serum creatinine was 121 μ mol/L before and 136 μmol/L after surgery (P =0.06).After a period of 22-47 months' follow-up,no patient had local recurrence or metastasis.Conclusions NSS can be safely performed and provide effective oncologic outcomes for selective patients with clinical T2 stage renal cell carcinomas.R.E.N.A.L nephrometry is an important factor and should be used to evaluate the feasibility of NSS.

Objective To explore the value of retinoblastoma binding protein 4 ( RBBP4 ) in diagnosing prostate cancer ( PCa).Methods From January 2015 to December 2015, the prostate tissue after prostatectomy were collected and the differentially expressed degree of RBBP4 protein was analyzed in PCa and adjacent tissues by 2D-DIGE technology.The RBBP4 score of prostate tissue chip which contains 3 normal prostate tissues, 7 cancer adjacent normal prostate tissues, 50 adenocarcinoma and 20 hyperplasia tissue was checked by immunohistochemistry( IHC).In 50 patients with PCa, 4 cases were less than 60 years old and 46 cases were more than 60 years.In those patients, the Gleason scores were less than 7 scores in 18 cases, and more than 7 scores in 30 cases.22 cases were confirmed less than Ⅱ stage, and 28 cases were confirmed more than Ⅲ stage.Finally, the RBBP4 IHC score and the clinic-pathological parameters such as age, Gleason score and clinical stage of PCa patients were analyzed together.Results We found that the protein of RBBP4 increased by 2.15 times in PCa tissues compared to adjacent tissues by using 2D-DIGE technology( P=0.008).The expression of RBBP4 was higher than that in benign tissues by IHC ( F=43.972,P=0.000).And the expression of RBBP4 was positive correlation with Gleason score( t=5.589, P=0.000) and clinical stage(t=5.620,P=0.000), but was negative correlation with age(t=1.125,P=0.266).Conclusions The detection of RBBP4 can help to separate PCa from benign tissues.The overexpression of RBBP4 might result in the rapid growth of malignant cells.It may have certain value in determine the clinical staging and pathological grading of PCa.

Objective To explore the role and clinical significance of GSTM 3 ( glutathione S-trans-ferase mu 3) expression in prostate cancer (PCa). Methods We had used the two-dimensional fluores-cence difference gel electrophoresis ( 2D-DIGE) and mass spectral analysis to further verify the microarray data of mRNA expression profiling discovered .GSTM3 mRNA level was detected by Rael-time Quantitative PCR ( RT-QPCR) in 28 pairs of prostate cancer tissue and benign tissue .The relationship of GSTM 3 level with the serum PSA level and the clinical feature of PCa were analyzed . Results In 2D-DIGE study, we found that the expression of GSTM 3 protein in adjacent tissues was significantly higher than that in PCa tis-sues (P<0.05).RT-QPCR results showed that GSTM3 in adjacent tissues (8.12±0.51) was significantly higher than that in PCa tissues (7.18±0.54) (P<0.05).There was no significant difference of GSTM3 ex-pression in different serum PSA packets ( P>0.05) and prostate cancer clinical pathological parameters ( P>0.05). Conclusions GSTM3 expression is down-regulated in PCa tissues, and we may identify PCa by detecting the GSTM 3 expression .

Objective To evaluate the effects and complications of percutaneous nephrolithotomy (PCNL) in the treatment of renal stone after repeated extracorporeal shockwave lithotripsy (ESWL).Methods Forty-four patients who had a history of repeated ESWL (treatment group) and 50 patients with-out surgical intervention (control group) were submited to PCNL,and clinical data was documented in details and analyzed.Results The time to establish access in treatment group and control group was (11.8 ± 4.1) min and (10.9 ± 2.5) min,respectively,and there was no significant difference (t =1.308,P =0.194).The time to extract stone in both groups was (92.0 ± 13.5) min and (66.6 ± 17.6) min,respectively,and there was significant difference (t =7.776,P =0.000).The operative time in treatment group was (113.9 ± 12.0) min,which was longer than that in control group with (87.6 ± 13.6) min (t =8.354,P =0.000).The clearance in both groups was 81.8％ and 94.0％,and there was no significant difference (x2 =3.361,P =0.067).The was no death or other severe complication in both groups.Conclusions The operation time in treatment group was longer than that in control group,and there was no significant difference in clearance and complication rate.Thus it was safe and effctive to perform PCNL in these patients with a history of failed repeated ESWL.