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Objective:To explore the correlation between CCAAT enhancer binding protein beta (CEBPB) expression and clinical characteristics in ccRCC, and to investigate the effect of CEBPB on proliferation and invasion of ccRCC cells.Methods:Between March 2020 to December 2020, the transcriptome and clinical data of 537 ccRCC cases were downloaded from TCGA database, and the correlation of CEBPB expression with clinical characteristics of ccRCC were analyzed. Univariate and multivariate Cox regression analysis were used to determine the effect of CEBPB expression on the prognosis of ccRCC patients. The correlation between CEBPB expression and immunocyte infiltration in ccRCC was investigated via TIMER database. The expression levels of CEBPB mRNA and protein in human renal tubular epithelial cell line HK2 and ccRCC cell lines (Caki-1, ACHN, 786O, 769P and A498) were determined by real-time PCR and western blot, respectively. After transfected with NC siRNA or CEBPB siRNA for 48 h, the proliferation and invasion of ACHN cells and 786O cells were determined by using MTT assay and invasion assay, respectively.Results:TCGA databases analysis revealed that, compared with normal kidney tissue, the expression of CEBPB mRNA in ccRCC was up-regulated by 2.55-fold ( P<0.05). CEBPB expression was positively correlated with age, tumor grade, tumor stage, lymph node metastasis and distant metastasis ( P<0.05). The tumor grade ( HR=1.703, P=0.040), tumor stage( HR=1.773, P=0.026), distant metastasis ( HR=3.080, P<0.001) and the high expression of CEBPB ( HR=1.874, P=0.003) were independent poor prognostic factors for ccRCC patients. The analysis results by using TIMER database showed that CEBPB expression was positively correlated with infiltrating levels of B cells (Rho=0.168), M2 macrophages (Rho=0.373), Tregs (Rho=0.348), neutrophils (Rho=0.194), and natural killer T cell (Rho=0.421) in ccRCC. The expression level of CEBPB mRNA in Caki-1, ACHN, 786O, 769P and A498 cells was (9.43±1.25)-fold, (5.44±0.82)-fold, (4.50±0.52)-fold, (4.88±0.73)-fold and (7.50 ± 1.04)-fold of HK2 cells, respectively. The expression level of CEBPB protein was (6.22±0.45)-fold, (5.84±0.85)-fold, (6.51±0.55)-fold, (6.23±0.62)-fold and (3.84±0.45)-fold of HK2 cells, respectively ( P<0.05). MTT assay showed that the proliferation rates of ACHN cells and 786O cells at 24, 48, 72, 96 h were (98.4±1.7)% and (99.0±1.4)%, (97.8±2.1)% and (98.5±1.5)%, (101.3±1.2)% and (97.6±1.7)%, (97.5±2.0)% and (99.1±1.3)% in NC siRNA group, and (68.8±5.8)% and (79.5±6.2)%, (57.9 ± 6.1)% and (70.8±5.1)%, (50.9±4.6)% and (66.8±4.9)%, (43.2±5.0)% and (60.5±5.3)% in CEBPB siRNA group. Compared with NC siRNA group, the proliferation activity of ACHN cells and 786O cells was significantly inhibited in the CEBPB siRNA group ( P<0.05). Cell invasion assay showed that the invasion activity of ACHN cells and 786O cells were (95.0±5.2)% and (97.3±4.4)% in NC siRNA group, (35.2±5.4)% and (26.7±3.3)% in CEBPB siRNA group, respectively ( P<0.05). Compared with NC siRNA group, the invasion activity of ACHN cells and 786O cells were significantly inhibited in the CEBPB siRNA group ( P<0.05). Conclusions:CEBPB was highly expressed in ccRCC, which was closely related to the prognosis and immunocyte infiltration of ccRCC patients. Silencing the expression of CEBPB significantly inhibited the proliferation and invasion of ccRCC cells
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Objective@#To observe the clinical efficacy of Ningmitai capsules, a traditional Chinese medicine using for clearing heat and dampness, in the treatment of residual fragments and postoperative complications following lithotripsy for upper urinary stones.@*Methods@#During October 2016 and March 2018, patients from Wuhan 1st Hospital, Wuhan 2nd Hospital, Wuhan 3rd Hospital, and Wuhan Puai Hospital having upper urinary residual fragments following minimally-invasive stone treatment were randomly assigned to control group and Ningmitai group with a proportion of 1∶3. The patients in control group were treated with antibiotics or sodium diclofenac suppository on demand, while patients in Ningmitai group took additional Ningmitai capsule orally (4 capsules per time, 3 times per day). The observation was started when a patient was enrolled in this study and continued for a maximum of 12 weeks or until stone-free status. During the observation, the stone expulsion time, stone-free time, stone-free rate were observed, and the difference in curative effect between the two groups on postoperative complications such as pain and infection were compared. Statistical analysis was done using t-test or χ2 test by GraphPad Prism 5 software.@*Results@#Totally 269 cases enrolled in this study. Eighty-six patients were from Wuhan 1st Hospital, 69 patients from Wuhan 2nd Hospital, 58 patients from Wuhan 3rd Hospital, 56 patients from Wuhan Puai Hospital, respectively. There were 66 cases in control group and 203 cases in Ningmitai group. The residual fragments expulsion time in Ningmitai group was significantly earlier than that in control group ((4.5±0.4) days vs. (7.5±1.3) days, t=2.877, P=0.004), the residual fragments clearance time in Ningmitai group was significantly shorter than that in control group ((13.6±1.0) days vs.(25.6±3.8) days, t=4.252, P=0.000), and the stone-free rate within 4 weeks post operation in Ningmitai group was significantly higher than control group (91.6% vs. 68.2%, χ2=22.57, P=0.000). After 12 weeks of treatment, the total effective rate of control group was 89.4%, and the total effective rate of Ningmitai group was 99.5%, with statistically significant difference (χ2=17.65, P=0.000). The proportion of caregivers that offered analgesia in Ningmitai group was significantly lower than that in control group (16.3% vs. 30.3%, χ2=6.212, P=0.013), the recovery rate of routine urinalysis following 4 weeks′ treatment was significantly higher in Ningmitai group than that in control group (88.2% vs.71.2%, χ2=10.67, P=0.001). No obvious adverse effects were observed in both groups.@*Conclusions@#Ningmitai capsule can facilitate the stone passage and increase the stone-free rate in the treatment of residual fragments and postoperative complications of upper urinary stones. It is also helpful for the prevention and treatment of postoperative pain, infection and other complications.
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Objective The aim of this research was to produce apparatus of urinary diversion using silk-fibroin loading rab-bit adipose stem cells,and assess the effect of urinary diversion in a rabbit model.Methods Adipose stem cells were obtained and cultured in vitro,and flow cytometry analysis was performed to determine the adipose stem cells.These cultured adipose stem cells were used to seed on the silk-fibroin scaffolds,and after being incubated in the conditioned medium for 7 days,the a-bove compounds were made into apparatus of urinary diversion.This apparatus of urinary diversion was implanted into 20 rab-bits with radical cystectomy to develop urinary diversion.Five rabbits from each experimental group were euthanized at the spe-cific time points(1,2,3,4 months postoperatively),and the implants were harvested for histological and immunohistochemical a-nalysis.In the control group,silk-fibroins with unseeded cells(only silk-protein scaffolds)were also made into apparatus of uri-nary diversion and then used as urinary diversion on another 5 rabbits with the same process.Results Rabbits adipose stem cells were isolated and cultured successfully,and determined by flow cytometry.The silk-fibroin scaffolds were synthesized suc-cessfully.All rabbits were alive in the experimental group until the time of sacrifice.Histological and immunohistochemical anal-ysis showed multilayer uroepithelium coverage in the luminal surface of apparatus of urinary diversion,and as time went on,epi-thelial layers increased continuously.In the control group,all animals were dead within 3 weeks,and urine leakage,severe in-flammatory reaction and tissue destruction were found by autopsy.Conclusion The present experiment has successfully used silk-fibroin loading rabbit adipose stem cells to construct apparatus of urinary diversion,and demonstrates the feasibility of this kind of apparatus for urinary diversion in a rabbit model,which provides some experimental basis for clinical applications.
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Objective To explore the saffety and efficacy of super-selective prostate artery embolization (PAE) combined with TURP (transurethral resection of prostate) as an alternative method for patients with severe large BPH (＞ 80 ml).Methods From March 2015 to June 2017,a total of 40 patients with large benign prostatic hyperplasia who failed in medical treatment were selected for PAE combined with TURP (18 cases)and TURP (22 cases).In the PAE combined with TURP group,the mean age was (75.0±8.7) years (ranging60-88 years) and the mean prostatic volume was (111.0 ±23.3) ml,ranged from 83 to 145 ml).The international prostate symptom score (IPSS),quality of life (QOL),maximal t rine flow rate (Qmax) and postvoid residual urine(PVR) were(25.2 ±3.6),(5.1 ± 1.0),(6.4 ± 2.3) ml/s and (107.7 ± 32.6) ml,respectively.In the TURP group,the mean age was (76.0 ± 6.9) years (ranging 62-85 years) and the mean prostatic volume was (107.5 ±27.4) ml,ranged from 80 to 150 ml).The IPSS,QOL,Q andPVRwere(24.3±4.2),(4.9 ±0.9),(6.7±2.2)ml/s and (106.6±32.2)ml,respectively.Clinical data of all of patients were analyzed retrospectively,including operative time,estimate blood loss,weight and efficacy of resected tissue,time of continuous bladder irrigation and catheterization,IPSS,QOL,PVR,Q and postoperative complications.Results There were significant differences in the operative time [(75.8 ± 25.1) min vs.(103.2 ± 27.7) min],estimate blood loss [(122.8 ± 33.9) ml vs.(447.6 ± 36.0) ml],weight of resected tissue [(99.9 ± 24.2) g vs.(82.9 ± 15.5) g],efficacy of resected tissue [(76.9 ± 20.7) g/h vs.(41.7 ± 14.2) g/h],continuous bladder irrigation time [(1.4 ± 0.5) d vs.(2.4 ± 0.8) d] and catheterization time [(2.2 ± 0.4) d vs.(3.4 ± 0.6) d] between PAE combined TURP group and TURP group (P ＜ 0.05).The postoperative complications of PAE combined TURP group and TURP group were included secondary hemorrhage (0 case vs.3 cases),secondary TURP (0 case vs.3 cases),temporary urinary incontinence (2 case vs.4 case),urinary tract infection (1 case vs.2 case).After 1-year follow up,the IPSS,QOL,Qmax and PVR of PAE combined TURP group and TURP group were (6.7 ±1.5)and(6.9± 1.5),(2.3 ±0.5) and(2.3 ±0.6),(15.6 ±2.3) ml/s and(15.0 ±2.1) ml/s,(32.8±6.5) ml and(32.3± 8.4)ml,respectively.Both goups were found to have significantly improved in IPSS,QOL,Q and PVR,as compared with preoperative indexes,respectively (P ＜ 0.05).However,there was no significant difference in those indexes between two groups (P ＞ O.05).Conclusions PAE combined TURP could be used a safe and effective therapy for treating patients with LUTS due to large volume (＞ 80 ml) BPH.It has been a priority in less blood,more efficient of resected tissue and less postoperative complications.
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Objective To study the expression of long non-coding RNA (lncRNA)-urothelial carcinoma associated 1 (UCA1) and miR-34b in bladder cancer and its correlation to the clinicopathologic features of bladder cancer.Methods Between January 2011 and October 2012,the expression of UCA1 and miR-34b in 5 bladder cancer cell lines (T24,BIU-87,EJ,T24-MMC,T24-ADM) and 1 normal bladder cell lines (SV-HUC-1) were measured by real-time reverse transcription-polymerase chain reaction (RTPCR).Meanwhile,the 56 bladder cancer specimens and paraneoplastic normal bladder tissues,which diagnosed by pathology were collected from bladder cancer patients undergoing radical resection of bladder.Among them,41 cases were male and 15 cases were female.The mean age was (68.4 ± 7.5)years old,range 52 to 78 years.43 cases were older than 65 years old,and 13 cases were less than 65 years old.The pathological classification included non muscle-invasive bladder cancer (NMIBC) 18 cases,muscle-invasive bladder cancer 38 cases;low grade papillary urothelial carcinoma 22 cases,high grade papillary urothelial carcinoma 34 cases;12 cases were primary lesion,the other 44 cases were diagnosed as tumor recurrence.Real-time RT-PCR was performed to analyze the expression of UCA1 and miR-34b.Results The relative expression levels of UCA1 in the normal bladder cell lines (SV-HUC-1) and 5 bladder cancer cell lines (T24,BIU-87,EJ,T24-MMC and T24-ADM) were (0.0675 ± 0.0133),(0.2934 ± 0.0531),(0.4246 ± 0.0650),(0.4206 ± 0.0826),(0.6472 ± 0.0875) and (0.7165 ± 0.1032),respectively (P ＜ 0.05).Moreover,the expression levels of UCA1 were up-regulated in 2 drug resistant bladder cancer cells lines T24-MMC (0.6472 ± 0.0875)and T24-ADM (0.7165 ± 0.1032),as compared with the T24 bladder cancer lines (0.2934 ± 0.0531),respectively (P ＜ 0.05).However,the expression levels of miR-34b in 5 bladder cancer cell lines [T24 (0.1600 ± 0.0455),BIU-87 (0.1720 ± 0.0658),EJ (0.1150 ± 0.0352),T24-MMC(0.0576 ± 0.0087),T24-ADM (0.0510 ± 0.0125)] were decreased (P ＜ 0.05),as compared with normal bladder cell lines SV-HUC-1 (0.6384 ± 0.1083).Moreover,the expression levels of miR-34b were down-regulated in 2 drug resistant bladder cancer cells lines T24-MMC (0.0576 ± 0.0087) and T24-ADM(0.0510 ± 0.0125),as compared with the T24 bladder cancer lines T24 (0.1600 ± 0.0455),respectively (P ＜ 0.05).The relative expression levels of UCA1 and miR-34b in bladder cancer tissues and paraneoplastic normal bladder tissues were (0.4225 ± 0.0714) vs.(0.0532 ± 0.0192) and (0.0340 ± 0.0134)vs.(0.5643 ±0.0616),respectively (P ＜0.05).Statistical correlation analysis showed that UCA1 to be significantly negative correlated with miR-34b in bladder cancer specimens(r =-0.54,P ＜ 0.05).The high level of UCA1 and low level of miR-34b were significantly correlated with tumor malignant grade,invasiveness and recurrence.The 3-year overall survival rate (OS) in UCA1 (+)/miR-34b(-) group (27.6％) were significantly worse compared with non UCA1 (+)/miR-34b (-) group (73.7％).Conclusion High expression of UCA1 and low expression of miR-34b were associated with the occurrence and development of bladder cancer.
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The anti-tumor activity of curcumin against androgen-independent prostate cancer cells in vitro and the possible mechanism were investigated. After curcumin treatment, the effect of curcumin on the proliferation of prostate cancer PC-3 cells was assessed by CFSE staining. Flow cytometery (FCM) was performed to analyze the cell cycle and the induction of apoptosis of tumor cells. A luciferase reporter gene assay was used to determine the effects of curcumin on the activities of intracellular NF-κB and AP-1 signaling pathways. The results showed curcumin could effectively inhibit the proliferation of PC-3 cells in vitro (P<0.05). Cells were arrested at G(2)/M phase. After curcumin treatment, the percentage of apoptotic cells was significantly higher than in control group (P<0.05). The results of the luciferase assay revealed that curcumin selectively inhibited the activities of the NF-κB and AP-1 signaling pathways in PC-3 cells significantly. It was suggested that curcumin could exert anti-tumor activity against androgen-independent prostate cancer cells in vitro by inhibiting cellular proliferation and inducing apoptosis, which was probably contributed to the inhibition of transcription factors NF-κB and AP-1.