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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:Mutation in the gap junction beta 6 (GJB6) gene has been reported to be associated with an autosomal dominant disorder hidrotic ectodermal dysplasia (HED), characterized by congenital nail clubbing, alopecia and palmoplantar keratoderma. The aim of this study is to investigate relationship between genetic mutation in GJB6 and HED in an affected Chinese family. Methods:We selected a Chinese HED family consisting of a total of 17 individuals including 8 HED patients (5 males and 3 females). The whole coding region of GJB6 was amplified by polymerase chain reaction and sequenced. Results:Sequence analysis identified a heterozygous missense mutation c.31G>A (p.G11R) in GJB6 gene of affected individuals, but not in healthy individuals. Conclusion:A c.31G>A (p.G11R) missense mutation in GJB6 gene is the genotypic characteristic for HED in Chinese population.
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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.
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Objective To determine the association of mutations in aldosterone synthase (CYPllB2)and 11 beta-hydroxylase(CYP11B1)genes with primary aldosteronism(PA).Methods Five mutations of CYP11B2 and CYP11B1 genes were analyzed in patients with PA and normal population.Among them,intron 2 was detected by 2 independent PCR reactions,and the others were analyzed using Taqman probes.The Haploview 4.0,SNPassoc 1.5-3 and Haplo.stats 1.3.8 were used to analyse the association between polymorphisms and PA.Results All the selected mutations were successfully genetyped.Only rs64lO allelic frequencies in patients with aldosterone-producing adenoma (APA)and idiopathic hyperaldosteronism(IHA)were significantly different with those in controls (P＜0.05).There was a relative excess of AA homozygotes and AG heterozygotes of rs6410 allele in APA group compared with control group(P＜0.01).There were significantly different genotypes AA and AG of rs6410 allele between patients with IHA and controls only after adjusted for age,gender,eeptible haplotype AAAWT was identified to be significantly associated with APA(OR=1.44,95%CI 1.19-1.76).Three susceptible haplotypes AAAWT,AGGWT and AGAWC were identified to be significantly associated with IHA(OR=1.55,95%CI 1.23-1.96;OR=1.49,95%CI 1.17-1.89;OR=1.40,95%CI 1.04-1.88).In contrast,1 protective haplotype GGAWT showed significant difference between patients with APA and controls(OR=0.73,95%CI 0.55-0.97).Conclusion There is a significant association between genetic variations in CYP11B2 and CYP11B1 genes and genetie predisposition to PA.
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Objective To investigate the in vitro effects of bladder cancer cell proliferation after silencing Notch1 gene. Methods The siRNA eukaryotic expression vector of Notch1 (psiRNA1)was constructed and transfected into bladder cancer cell lines T24 and BIU-87. Methabensthiazuron (MTT) and flow cytometry (FCM) assays were used to detect bladder cancer cells line growth, cell cycle and apoptosis after the transfection. RT-PCR and Western blotting were used to determine the expression changes of Notch1 in these cell lines. Results After transfection for 72 h, the rate of G0/G1 phase cells inceased from (23.89±1.32) % to (80.13±2.69)% in T24 cell line, and increased from (24.63±1.68)% to (69.44±2.41)% in BIU-87 cell line (both P<0.05). In addition, apop-totic cell index in T24 and BIU-87 cell lines increased from (1.28±0.14)% to (13.75±1.23)%, from (1.01±0.27)% to (8.72±1.01)%, respectively(both P<0.05). The growth of T24 and BIU-87 cell lines was obviously inhibited 24 h after the transfection, and the inhibitory effects lasted until 96 h after the transfection. Notch1 mRNA and protein significantly downregulated after transfection compared to the control(P<0.05). Conclusions Silencing Notch1 expression can inhibit the prolif-eration of bladder cancer cell lines. Notch1 gene might act as a tumor gene in bladder cancer.
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Objective To validate a new standardized training program for urological surgeons to improve their laparoscopic surgical skills. Methods The laparoscopic surgical training program was carried out by using the traditional mechanical simulators and animal models. Thirty-three trainees participated the urological laparoscopic surgical training program. Initially, the novices were assigned to practise basic laparoscopic skills step by step on the simulator with fixed trocar positions. After a period of basic training, they were allowed to practise on animal models for some particular proce-dures. Results All trainees (33/33, 100.0%) participated were able to perform all basic techniques skillfully and completed laparoscopic anastomosis accurately after the training. The time required for performing the partial nephrectomy, dismembered pyeloplasty and ureteral reimplantation on animal models declined from 64.0±18.4, 127.54±17.5 and 75.84±11.6 min at the beginning to 30.94±3.8, 65.2±7. 5 and 37.7±7.2 min after practicing these procedures 8 times (P<0.01). They could un-derstand the crucial procedures of the laparoseopic surgeries after 6 to 8 special trainings on animal models. Fifteen trainees (15/33, 45.5%) had started to carry out laparoscopic surgeries after-finish- ing the training program. Conclusions Our program enables the participants to improve their techniques in complicated laparoscopic surgeries. The challenging parts of reconstructive laparoscopic surgeries such as laparoscopic dismembered pyeloplasty can be taught by using animal models. This program could be incorpo-rated easily by all urological departments developing a laparoscopie surgical training program.