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Objective To evaluate the short-term outcome of the Kelly procedure for bladder exstrophy-epispadias complex.Methods Kelly procedure was performed in 5 cases from December 2015 in our institute. Retrospective analysis was done including the surgery and follow up.Results Two bladder exstrophy boys had histories of primary bladder closure without bladder neck reconstruction during the neonatal period. Another three epispadias boys had sever urethral defects with full incontinence. All cases undertook Kelly procedure and were followed up for 7.8 months (2-12 months). No severe complications such as glanular necrosis, urethral stricture and urethrocutaneous fistula was observed. Four cases were fully continent and one epispadias case had stress incontinence.Conclusions Kelly procedure produced satisfactory urinary continence and phallic cosmetics in bladder exstrophy-epispadias complex in children. Proper identification of the pudendal neurovascular bundle and repair of the external sphincter by levatorplasty help improve the outcome.
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Objective To evaluate the feasibility and outcomes of robotic-assisted laparoscopic pyeloplasty in children .Methods A retrospective study was performed in patients who underwent robotic-assisted laparoscopic pyeloplasty ( Anderson-Hynes ) at our institution between January 2014 to August 2014.Totally 6 boys were diagnosed as left ureteropelvic junction obstruction depending on the symptoms and radiographic studies .The mean age was 9 years ( range 4 -12 years ) .Results The procedure was performed successfully without conversion to open surgery in all of the cases .Mean operative time was 216 min (range 175-269 min), with a mean robotic anastomosis time of 45 min (range 30-60 min).Mean estimated blood loss was less than 15 ml.The mean hospitalization was 4.5 days.Mean follow-up period was 10 months ( range 7 -14 months ) .There were no perioperative complications , and recovery was uncomplicated (without recurrence, pyelonephritis, nephrarctia) in all of the patients.Conclusion Robotic-assisted laparoscopic pyeloplasty can be safely performed in children older than 4-year-old with ureteropelvic junction obstruction .
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Ranpirnase (onconase, ONC) is a new drug, with weak RNase activity and strong cytotoxicity to various tumor cells in vitro and in vivo. This study is to obtain recombination onconase (rONC) with high bioactivity. Based on the codon preference of Pichia pastoris, we designed and synthesized the gene according to cDNA sequences of ONC and the α mating factor's prepeptide. We screened positive clones after transforming the recombination plasmids into P. pastoris X-33, GSS115 and SMD1168. We screened the best combination of seven different vectors and host strains. Moreover, we optimized culture condition in shake flasks and 10 L bioreactor, and purified rONC from the supernatant after inducing it with 0.25% methanol by aqueous two-phase extraction coupling G50 molecular exclusion method. The highest rONC production was 13 mg/L in pPICZα-A/X-33/ONC combination under the condition of pH 5.5 and 23 degrees C in shake flasks for 7 d; and that the highest rONC production was 180 mg/L when the induction is performed in the lower basic salt medium with pH 5.5 in the 10 L bioreactor for 7 d. The yield of rONC is more than 90% at a purity of above 95%. rONC can kill various tumor cells in vitro. The expression and purification of rONC would be useful for further investigation of this new drug.
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Humans , Antineoplastic Agents , Metabolism , Bioreactors , Cell Line, Tumor , Codon , DNA, Complementary , Genetic Vectors , Pichia , Metabolism , Recombinant Proteins , RibonucleasesABSTRACT
Objective To observe the expression and mechanism of monocyte chemoattractant protein-1 (MCP-1) in interstitial cystitis (IC) rats.Methods Twenty weight of 250-300 g of female SD rats were divided into IC group (n =10) and control group (n =10).IC group were treated by transurethral instillation with 10 mg/ml protamine sulfate (PS) 1 ml reserved for 45 min,and then instillation with 750 ug/ml of lipopolysaccharide (LPS) 1 ml reserved for 30 min.The same operations were repeated after 24 hours,and the rats were killed obtaining the bladder tissue and urine after three days.Control group was given PBS solution perfusion.MCP-1 and histamine (HA) expression levels in the rat bladder tissue and urine were detected by ELISA.The inflammation of bladder tissue was observed and inflammatory score was used by HE staining.MC degranulation count was used by MC special staining.MCP-1 expression and distribution in bladder tissue was observed by immunohistochemical method.The relationship between the MCP-1 and MC was detected by immunofluorescence method.Results By ELISA,the expression levels of MCP-1 and HA in the bladder tissue and urine in IC group were significantly increased compared with control group (P ＜0.01 ).More inflammatory cell infiltration in the bladder mucosa,edema mucosa,congestion and hemorrhage were seen by HE staining.The inflammatory score in IC and control group were (76.5 ±9.8) and (18.5± 9.8)/field (P ＜ 0.01 ).With MC special staining,degranulation MC count in IC and control group were (6.4±3.1 ) and (0.7 ±0.3)/field (P ＜0.01 ),and the degranulation in the bladder tissue of IC group was significantly higher than control group (P ＜ 0.01 ).MCP-1 has a higher expression in the bladder epithelium,and more MCP-1 were found gathering round MC surface by immunofluorescence.Conclusions MCP-1 is highly expressed in IC rats,and could induced activation of MC,which could release HA,aggravating the pathological process of inflammatory and fibrosis in IC.