Objective To observe the effects and study the underlying mechanism of siRNA targeting PARP1 on the proliferation of androgen independent prostate cancer PC3 cell line.Methods Three specific siRNA sequences targeting PARP1 were designed and synthesized.And two sequences which had better interfering effect on the expression of PARP1 were evaluated and selected through lipofectamine transfection,RT-PCR and Western Blot.The effect of PARP1 silencing on the proliferation of PC3 cells was observed with MTS assay and the levels of the phosphorylation of Akt and GSK3β were detected by Western Blot.Results Compared to the blank control group,the transfected group with the negative control sequence had no significant impact on the expression of PARP1,however the transfected group with siRNA-1706,-2003 or-2907 could significantly suppress the mRNA and protein expression of PARP1.The mRNA inhibition rate reached to(52.07 ± 4.65)％,(44.38 ± 9.15)％ and(22.05 ± 6.65)％,respectively;and the protein inhibition rate reached to(86.86 ± 4.94)％,(83.30 ± 7.18)％ and(63.05 ± 10.19)％,respectively.The siRNA-1706 and-2003 could significantly inhibit the proliferation of PC3 cells;the inhibition rate was(38.93 ± 3.87)％ and(34.93 ± 1.21)％.And they also could down-regulate the intracellular levels of phosphorylated Akt and GSK3β in PC3 cells.Conclusion PARP1-targeted siRNA can significantly suppress the expression of endogenous PARP1 and inhibit the proliferation of PC3 cells,which is related to the inhibition of Akt activity and the activation of GSK3 β.

Objective To study the expression of PARP and its effect on apoptosis and proliferation in androgen dependent prostate cancer LNCaP cells.Methods The expression of PARP in a LNCaP cell line with or without PARP inhibitor 5-AIQ was measured by Western blot.The effect of 5-AIQ on the proliferation of LNCaP cells was analyzed with MTS assay,and flow cytometry analysis was performed to assess the apoptosis of LNCAP cells induced by 5-AIQ treatments.Results The protein expression of PARP in LNCaP cell line decreased to 65.3％ or 22.4％ in the 5-AIQ treatment group (500 μmol/L or 1000 μmol/L) respectively,which was obviously suppressed compared with the blank group ( P ＜ 0.05 ).When 5-AIQ was applied,from 200 to 1000 μ mol/L,the inhibition ratio of the proliferation of LNCaP cells was increased from (2.85±2.03)％ to (41.23 ±5.42)％,(2.85 ±2.03)％ to (41.23 ±5.42)％,or (25.67 ±0.63 ) ％ to (65.81 ± 1.62 ) ％ after treatment for 24 h,48 h and 72 h.The growth of LNCaP cells was significantly inhibited compared with the blank group in a dose-time-effect relationship ( P ＜ 0.05 ).Flow cytometry analysis showed that the number of apoptosis cells in early,late and total phase induced by different doses (500 μ mol/L or 1000 μmol/L) of 5-AIQ after 48 h were 23.6％,4.6％,28.2％ and 31.8％,6.3 ％,38.1％ respectively,which was significantly higher than those in the blank group ( P ＜ 0.05 ).Conclusions The expression of PARP in LNCaP cell line was suppressed by PARP inhibitor 5-AIQ,which can both inhibit proliferation and induce apoptosis of the LNCaP cell line.PARP is expected to become a new therapeutic target for prostate cancer in the future.

ObjectiveTo investigate the clinical value of fluorescence in situ hybridization (FISH) in the diagnosis of urothelial carcinoma.MethodsFrom September 2010 to October 2011,morning voiding urine of 27 patients with suspected urothelial carcinoma was collected for FISH examination.The results of FISH examination were compared with the results of pathological examination by endoscopic biopsy.Both sensitivity and specificity were compared respectively,and the cost of each kind of examination was also considered.ResultsOf 27 cases,pathological examination by endoscopic biopsy revealed 11 cases of urothelial carcinoma,FISH examination showed 9 cases of urothelial carcinoma,but only 7 cases in these 11 cases.The overall sensitivity of FISH examination was 63.6％ (7/11 ),the specificity was 87.5％(14/16).The cost per case of FISH examination (3100 yuan) was 3.1 times of pathological examination by endoscopic biopsy( 1000 yuan).ConclusionsIt showed that there is no advantage of FISH examination for diagnosis of urothelial carcinoma considering both the sensitivity and specificity,and the cost is also higher than that of pathological examination by endoscopic biopsy.It should be evaluated further when FISH examination is widely used in clinic.

Calcium oxalate (CaOx) stone is the main type, and its formation is closely related to the metabolism of oxalic acid and calcium. Gut Microbiome is normal microflora which settled in the human intestinal tract and plays an important role in regulating a variety of metabolism in the body. In the past, Oxalobacter formigenes in gut was a protective factor for the formation of CaOx stones. Recently, it has been found that the bacteria regulating oxalate metabolism were not limited to Oxalobacter formigenes. Gut Microbiome of CaOx stones formers is different from healthy people. It regulates the metabolism of oxalic acid in the body through the gut-kidney axis and affect the formation of CaOx stone. The purpose of this study is to describe the characteristics of intestinal flora in patients with CaOx stones, and to summarize its potential function in the formation of CaOx stones and its possible clinical application in the future.

OBJECTIVETo establish a high-performance liquid chromatography-mass spectrometry (LC/MS)-based method for efficient determination of melamine in urinary calculi in children taking melamine-contaminated formula milk powder.

METHODSLC/MS was employed to determine the contents of melamine in urinary calculi surgically removed from 17 children with a history of taking melamine-contaminated milk powder and in 4 samples of uric acid stone from adults. The positive ionization mode of electro-spray ionization source was used, and the limit of melamine determination was 0.1 mg/kg.

RESULTSNo melamine was detected in the 4 uric acid stone samples from adults. Melamine was detected in 4 samples of urinary calculi from the 17 children, with the concentration ranging from 0.8 to 64 mg/lkg.

CONCLUSIONLC/MS is simple and effective for detecting melamine in urinary calculi, which is helpful to the treatment and follow-up.

Chromatography, High Pressure Liquid ; methods ; Dairy Products ; adverse effects ; Female ; Food Contamination ; Humans ; Infant ; Male ; Mass Spectrometry ; methods ; Triazines ; analysis ; Urinary Calculi ; chemically induced ; chemistry

Objective:To investigate the characteristics of distribution and drug resistance of urinary bacteria in the mid-stream urine of patients with infectious stones.Methods:The retrospective study analyzed the clinical data of 254 patients with infectious stones in the First Affiliated Hospital of Guangzhou Medical University from September 2016 to September 2018. All patients were treated with PCNL. Overall, there were 101 male and 153 female patients, with the mean age of(51.5±12.3) years, and the mean stone burden of 1443.5(660.8, 2837.5) mm2. There were 58 (22.8%) patients with hypertension, 17(6.7%) patients with diabetes and 195(76.8%)with hydronephrosis. The mid-stream urine samples were obtained for bacterial culture and susceptibility test, and the results of urine culture and antimicrobial susceptibility were recorded and analyzed.Results:Of 254 patients involved in this study, 89(35.0%) were positive and 165 (65.0%) were negative for urinary bacterial culture of the mid-stream. The proportion of patients with positive urine bacterial culture of the mid-stream who had positive urine leucocytes, positive urine nitrite and postoperative pyrexia were 86.5%(77/89), 64.0%(57/89), 25.8%(23/89), respectively, which was higher than that of patients with negative urine bacterial culture of the mid-stream [50.3%(83/165), 14.5%(24/165), 14.5%(24/165), P<0.05]. Four teen kinds of bacteria were detected from the mid-stream urine, and the three bacteria with the highest detection rate in turn were Escherichia coli of 38.2%(34/89), Proteus mirabilis of 15.7%(14/89), and Pseudomonas aeruginosa of 11.2%(10/89). The results of this study showed that three common bacteria had high resistance to drug including Cefazolin, Cefuroxime, Cefuroxime ester, Ampicillin and Co-trimoxazole (all resistance rate>40%). The resistance rates of Escherichia coli and Pseudomonas aeruginosa to Ciprofloxacin and Levofloxacin were higher than or equal to 40%. The resistance rates of Escherichia coli and Proteus mirabilis to meropenem, imipenem, ertapenem, piperacillin/tazobactam and amikacin were all lower than 10%. In addition, the resistance rates of Escherichia coli to nitrofurantoin and tigecycline and Proteus mirabilis to tobramycin, aztreonam and cefoxitin were all less than 10%. The resistance rates of Pseudomonas aeruginosa to ceftazidime, cefepime, gentamicin and aztreonam were less than 10%. Conclusions:The highest detection rate of urinary bacteria in culture of the mid-stream with infectious stones are Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa, all of which showed high resistance to Ampicillin, Cotrimoxazole, and some cephalosporins. Escherichia coli and Pseudomonas aeruginosa showe high resistance to Ciprofloxacin and Levofloxacin, and all of the three bacteria have low resistance rates to some β-Lactamase inhibitor complex and carbapenems, suggesting a reference for clinical empirical medical treatment.

Overactive bladder (OAB) is the most bothersome symptom in lower urinary tract symptoms (LUTS). Current pharmacologic treatment aims to inhibit detrusor contraction; however, shows unsatisfied efficacy and high discontinuation rate. LIM kinases (LIMKs) promote smooth muscle contraction in the prostate; however, their function in the bladder smooth muscle remains unclear. Here, we studied effects of the LIMK inhibitors on bladder smooth muscle contraction and proliferation both