Objective To investigate the role of RNA-binding motif protein X-linked(RBMX)in regulating the proliferation,migration,invasion and glycolysis in human bladder cancer cells.Methods A lentivirus vectors system and RNA interference technique were used to construct bladder cancer 1376 and UC-3 cell models with RBMX overexpression and knockdown,respectively,and successful cell modeling was verified using RT-qPCR and Western blotting.Proliferation and colony forming ability of the cells were evaluated using EdU assay and colony-forming assay,and cell migration and invasion abilities were determined using Transwell experiment.The expressions of glycolysis-related proteins M1 pyruvate kinase(PKM1)and M2 pyruvate kinase(PKM2)were detected using Western blotting.The effects of RBMX overexpression and knockdown on glycolysis in the bladder cancer cells were assessed using glucose and lactic acid detection kits.Results RT-qPCR and Western blotting confirmed successful construction of 1376 and UC-3 cell models with RBMX overexpression and knockdown.RBMX overexpression significantly inhibited the proliferation,clone formation,migration and invasion of bladder cancer cells,while RBMX knockdown produced the opposite effects.Western blotting results showed that RBMX overexpression increased the expression of PKM1 and decreased the expression of PKM2,while RBMX knockdown produced the opposite effects.Glucose consumption and lactate production levels were significantly lowered in the cells with RBMX overexpression(P<0.05)but increased significantly following RBMX knockdown(P<0.05).Conclusion RBMX overexpression inhibits bladder cancer progression and lowers glycolysis level in bladder cancer cells by downregulating PKM2 expression,suggesting the potential of RBMX as a molecular target for diagnosis and treatment of bladder cancer.

Objective To investigate the role of RNA-binding motif protein X-linked(RBMX)in regulating the proliferation,migration,invasion and glycolysis in human bladder cancer cells.Methods A lentivirus vectors system and RNA interference technique were used to construct bladder cancer 1376 and UC-3 cell models with RBMX overexpression and knockdown,respectively,and successful cell modeling was verified using RT-qPCR and Western blotting.Proliferation and colony forming ability of the cells were evaluated using EdU assay and colony-forming assay,and cell migration and invasion abilities were determined using Transwell experiment.The expressions of glycolysis-related proteins M1 pyruvate kinase(PKM1)and M2 pyruvate kinase(PKM2)were detected using Western blotting.The effects of RBMX overexpression and knockdown on glycolysis in the bladder cancer cells were assessed using glucose and lactic acid detection kits.Results RT-qPCR and Western blotting confirmed successful construction of 1376 and UC-3 cell models with RBMX overexpression and knockdown.RBMX overexpression significantly inhibited the proliferation,clone formation,migration and invasion of bladder cancer cells,while RBMX knockdown produced the opposite effects.Western blotting results showed that RBMX overexpression increased the expression of PKM1 and decreased the expression of PKM2,while RBMX knockdown produced the opposite effects.Glucose consumption and lactate production levels were significantly lowered in the cells with RBMX overexpression(P<0.05)but increased significantly following RBMX knockdown(P<0.05).Conclusion RBMX overexpression inhibits bladder cancer progression and lowers glycolysis level in bladder cancer cells by downregulating PKM2 expression,suggesting the potential of RBMX as a molecular target for diagnosis and treatment of bladder cancer.

The clinical data of 120 patients admitted to our hospital with renal stones treated by flexible ureteroscopy (FURS) and the imaging-related data measured by virtual reality technology were retrospectively analysed. The results of the univariate analysis showed that stone surface area (S), renal pelvis volume (P), length of calyces funnel (L), pelvic calyceal height (H) and essence of stone (E) were closely related to stone-free rate. The H. L.P.E.S. score was constructed to predict stone-free rate after FURS based on the above factors, and the area under the receiver aperating characteristic curve for the H. L.P.E.S. and S. O.L.V.E. scoring systems was 0.921 and 0.754 respectively.The H. L.P.E.S. scoring system has higher predictive value.

Objective To establish S.O.L.V.E.nephrolithometry scoring system,and to evaluate value of S.O.L.V.E.scoring system for predicting the stone-free rate (SFR) of flexible ureteroscopy (FURS).Methods Five reproducible variables were included in S.O.L.V.E.scoring system,such as stone surface area (S),obstruction (O),length of calyces funnel (L),visible number of calyces (V) and essence of stone (E).Variables were measured based on preoperative non-contrast computed tomography of urography.Clinical data of 392 patients who underwent FURS for upper urinary tract stones in our department from January,2017 to Jnne,2018 were retrospectively analyzed.The total study population consisted of 258 male and 134 female patients.The mean age was (49.5 ± 12.6) years old,ranged from 15 to 85 years.There were 292 patients in stone-fiee group,including 197 male and 95 female patients.The average age was (49.2 ± 12.8) years old.37 patients had previous history of renal stone surgery.Median body mass index was 24.7 kg/m2 (18.1-29.0 kg/m2) and median value of preoperative serum creatinine was 72.5 μmol/L (48.9-84.8 μmol/L).The number of patients,whose stone located in the left side and right side were 155 and 137,respectively.The number of patients,whose stones located in ureter,renal pelvis,lower calyceal and non-lower calyceal were 19,16,87,170,respectively.There were 100 patients in non-stone free group,61 men and 39 women.The average age was (50.4 ± 12.0) years old.15 patients had previous history of renal stone surgery.Median body mass index was 25.0 kg/m2 (18.5-28.8 kg/m2) and median value of preoperative serum creatinine was 73.8 μmol/L (46.5-92.5 μnol/L).The number of patients,whose stone located in the left side and right side were 51 and 49,respectively.The number of patients,whose stones located in ureter,renal pelvis,lower calyceal and non-lower calyceal were 7,4,27,62,respectively.The correlation of S.O.L.V.E.scoring system and stone-free rate,postoperative hospital stay,surgical complications,operation time were analyzed.Receiver operating characteristic curves were drawn to detect predictive value of S.O.L.V.E.scoring system for SFR of FURS.Results All cases FURS were performed successfully and the SFR was 74.5％ (292/392).Among the variables of the S.O.L.V.E.scoring system in the stone-free group and the non-stone free group,item S were (82.6 ± 69.8) mm2 and (172.6±133.7)mm2,respectively.The item L were (12.7 ± 15.8) mm and (23.9 ± 15.3)mm,respectively.The item V were (0.6 ± 0.7) and (1.3 ± 0.8),respectively.The item E were (817.1 ± 285.5) HU and (902.4 ± 256.1) HU,respectively.The difference was statistically significant (P ＜ 0.01).The item O was (17.7 ± 10.9) mm and (19.3 ± 13.1) mm,respectively,no statistical significance was found (P =0.242).The mean score was 6.3 (ranging 4-11) in this c ohort.The patients were divided into low score (4-5) group,moderate score (6-8) group and high score (9-11) group due to S.O.L.V.E.scoring system,and the stone-free rates were 93.5％ (130/139),70.5％ (153/217) and 37.5％ (9/36),respectively (P ＜0.01).The operation time of low,moderate,andi high score group were (31.6 ± 10.9),(42.3 ± 18.3),and (58.0 ± 19.2) min,respectively.Additionally,the score was correlated with the operation time(P ＜0.01),but not with postoperative hospital stay (P =0.133),intraoperative bleeding (P =0.185) and postoperative infectious fever (P =0.839).In logistic regression model analysis,the stone surface area,length of calyces funnel,number of involved calyces were significantly correlated with SFR (P ＜ 0.01).The obstruction degree and essence of stone were not associated with SFR (P ＞ 0.05).The area under receiver operating characteristic curve of S.O.L.V.E.score was 0.782,higher than that of each variable in S.O.L.V.E.scoring system(S,O,L,V,E were 0.738,0.535,0.698,0.735,0.593,respectively).Conclusions The stone surface area,length of calyces funnel,number of involved calyces were significantly correlated with SFR.The S.O.L.V.E.nephrolithometry scoring system can predict SFR after FURS accurately,and provide assistance for making clinical decisions.

Objective: To study the reversal effect of cyclin-dependent protein kinase 4/6 (CDK4/6) inhibitor palbociclib (PAL) on the resistance of breast cancer MCF-7/doxorubicin (DOX) cells to DOX, and to explore its mechanism from the view of drug efflux transporter ATP binding cassette protein B1 (ABCB1). Methods: The effect of PAL on the sensitivity of parental MCF-7 and resistant MCF-7/DOX cells to DOX was detected by CCK-8 method. The effect of PAL on intracellular accumulation of DOX in MCF-7 and MCF-7/DOX cells was determined by FCM analysis. The MDCK II cell model stably transfected with ABCB1 gene was constructed, then the effect of PAL on the ABCB1-mediated efflux function of DOX was detected by drug transmembrane bi-direction transport assay. The effect of PAL on the expressions of ABCB1 mRNA and protein in MCF-7 and MCF-7/DOX cells was detected by real-time fluorescent quantitative PCR and Western blotting, respectively. Results: After treatment with 0.2 and 0.4 μmol/L PAL, the half maximal inhibitory concentration (IC50) values of DOX in MCF-7/DOX cells were decreased by 39% and 51%, respectively (both P < 0.05), but the IC50 value of DOX in MCF-7 cells had no obvious change. In MCF-7/DOX cells treated with 0.4 μmol/L PAL, the intracellular accumulation of DOX was increased by 82% (P < 0.05), but the concentration of DOX in MCF-7 cells did not change. PAL significantly inhibited the ABCB1 -mediated efflux of DOX, but had no obvious regulatory effect on the expressions of ABCB1 mRNA and protein in MCF-7 cells and MCF-7/DOX cells. Conclusion: PAL can reverse DOX-resistance of breast cancer cells by inhibiting the function of drug transporter ABCB1, which suggests that coadministration of PAL and DOX maybe improve the effiocacy of DOX in breast cancer and optimize the clinical treatment.

Objective The explore regulation mechanism of specific proteins c-kit of interstitial cells of Cajal in the pathogenesis of slow transit constipation in rats.Methods Slow transit constipation in rat model was duplicated by injections of morphine (25 mg/kg,45 d).The number and weight of fecal granule were assayed.Daily average number of fecal particles,quality,and the time for first black discharge were analyzed.At the same time,the c-kit and SCF expression were detected by Real-time PCR and Western blot.The changes of inflammatory factors such as IL-1β,IL-2 and IL-10 were also detected by ELISA.Results Daily average number of fecal particles in constipation group was decreased,quality was higher,and the first time for first black discharge was prolonged,which showed significant differences (P ＜ 0.05).The expression levels of c-kit and SCF were decreased significantly,and levels of inflammatory factors IL-1β,IL-2 and IL-10 were higher in constipation rats than the controls (P ＜ 0.05).Conclusion The pathogenic progress of slow transit constipation rats are regulated,which is also related with inflammatory reactions.

Objective The explore regulation mechanism of specific proteins c-kit of interstitial cells of Cajal in the pathogenesis of slow transit constipation in rats.Methods Slow transit constipation in rat model was duplicated by injections of morphine (25 mg/kg,45 d).The number and weight of fecal granule were assayed.Daily average number of fecal particles,quality,and the time for first black discharge were analyzed.At the same time,the c-kit and SCF expression were detected by Real-time PCR and Western blot.The changes of inflammatory factors such as IL-1β,IL-2 and IL-10 were also detected by ELISA.Results Daily average number of fecal particles in constipation group was decreased,quality was higher,and the first time for first black discharge was prolonged,which showed significant differences (P ＜ 0.05).The expression levels of c-kit and SCF were decreased significantly,and levels of inflammatory factors IL-1β,IL-2 and IL-10 were higher in constipation rats than the controls (P ＜ 0.05).Conclusion The pathogenic progress of slow transit constipation rats are regulated,which is also related with inflammatory reactions.

Objective To investigate the expression change of RACK1 ,Src and Bcl‐2 in gastric carcinoma tissue and adjacent carcinomatous tissue .Methods Eighty specimens of gastric carcinoma and adjacent carcinomatous tissues in our hospital from Au‐gust 1 ,2011 to February 1 ,2014 were collected .The immunohistochemistry staining and Western blotting methods were adopted to detect the expression of RACK1 ,Src and Bcl‐2 protein in gastric carcinoma and adjacent carcinomatous tissues ,and their correlation was analyzed and performed the statistical analysis by combining with the clinicopathological data .Results The immunohistochem‐istry staining and Western blotting detection displayed that the expression positive rate and expression level of RACK 1 in gastric carcinoma tissue were obviously lower than those in the adjacent carcinomatous tissue ,while the expression positive rate and ex‐pression level of Src and Bcl‐2 in gastric carcinoma tissue were obviously higher than those in the adjacent carcinomatous tissue ,the differences were statistically significant (P<0 .05) .The RACK1 expression in gastric carcinoma tissue was negatively correlated with the Src and Bcl‐2 expression(r= -0 .632 ,-0 .754 ,P<0 .01) ,while Src had no obvious correlation with Bcl‐2 protein(r=0 .217 ,P>0 .05) .Conclusion The expression of RACK1 in gastric carcinoma tissue is significantly decreased ,while the expres‐sions of Src and Bcl‐2 are increased .

BACKGROUND:The human telomerase reverse transcriptase gene (hTERT) transfected into target cel s can play an important role in target cel proliferation and differentiation by increasing telomerase activity and maintaining telomere length. OBJECTIVE:To explore the effect of hTERT transfection on telomerase activity and biological characteristics of precartilage stem cel s culured in vitro. METHODS:Precartilage stem cel s cultured in vitro were subjected to hTERT gene transfection via a retrovirus vector pLXSN. Meanwhile, control and negative control groups were set up. After transfection, TRAP-ELISA assay was used to detect telomerase activity;RT-PCR and western blot employed to detect hTERT mRNA and protein expressions;cel counting kit-8 used to detect cel proliferaiton based on cel growth curve;and flow cytometry adopted to detect cel cycle and distribution. RESULTS AND CONCLUSION:The telomerase activity was significantly increased at 48 hours after hTERT gene was transfected into the precartilage stem cel s. After transfection of hTERT, hTERT mRNA and protein levels were significantly increased, the cel growth rate was significantly increased, the proportion of cel s at G 0/G 1 phase was decreased, and the number of S-phased cel s increased compared with the control group and negative control group. There were significant differences among the groups (P<0.05). In conclusion, hTERT transfection via retrovirus vector pLXSN can promote the proliferation of precartilage stem cel s in rats by increasing the telomerase activity.

Objective To investigate the effect of different pore sized hydroxyapatite for promoting bone vascularization in tissue engineering.Methods Male Wistar rats were randomly divided into three groups,named group A,B and C,which were im-planted hydroxyapatite bioceramics compositing 4 μg bone morphogenetic protein with different aperture of 200 -300,350 -450, 500-600 μm in the back subcutaneously.The size of each block was 5 mm×5 mm×1 mm in a weight about of 40.0 mg.After im-plantation,the animals were killed and the implants and the surrounding tissue were taken out at the first,second,third and forth week respectively.HE staining of histological analysis was used to detect the situation of local neovascularization.Results There was significant difference between second and third week in group A.Comparing the area of vascularization at different time points in group B and group C,there were significant difference in the comparison of intragroup (P <0.05 ).During the first week after surgery,there was only group C that had the area of vascularization.During the second and forth week after operation,the area of vascularization in group B and group C were significant higher than group A (P <0.05).The C group showed a great deal of new-born blood vessels and clear formation of bone trabeculae.Conclusion The hydroxyapatite bioceramics of 500-600 μm could better promote vascalarization of tissue engineering in bone.