Xu Zhang

Association of Low Expression of Notch-1 and Jagged-1 in Human Papillary Bladder Cancer and Shorter Survival

PURPOSEnWe investigated the expression of Notch receptors and ligands in normal bladder transitional epithelium and transitional cell carcinoma of the bladder. We also explored its clinical and pathological implications.nnnMATERIALS AND METHODSnThe expression of Notch-1 to 3, Jagged-1 and Delta-like-1 was detected respectively in 70 cases of bladder carcinoma, 10 of normal urothelium and the 2 cell lines T24 and BIU-87 using immunohistochemistry. Reverse transcriptase-polymerase chain reaction and Western blot were used to assay the expression level of Notch-1 and Jagged-1. The predictive value of this expression for prognosis was investigated by Kaplan-Meier curves and Cox proportional hazards analysis in a multivariate model.nnnRESULTSnAll 5 kinds of Notch factors were intensively stained in normal bladder transitional epithelium immunohistochemically but expression was significantly decreased in tumor tissues. Moreover, expression of the 5 genes in papillary tumors was lower than in invasive tumors but only Notch-1 and Jagged-1 showed a statistically significant difference. Postoperative disease-free survival time in patients with low Notch-1 plus Jagged-1 expression was significantly shorter than that in patients with other expression patterns in papillary tumors (p = 0.014). Multivariate Cox proportional hazards model analysis identified Jagged-1 expression as an independent prognostic factor for disease-free survival (RR 3.09, p = 0.011).nnnCONCLUSIONSnThe Notch family expression pattern in papillary bladder transitional cell carcinoma is different from that in invasive bladder transitional cell carcinoma. Low expression of Notch-1 as well as Jagged-1 is potentially a useful marker for survival in patients with papillary bladder transitional cell carcinoma.

The interaction of white spot syndrome virus envelope protein VP28 with shrimp Hsc70 is specific and ATP-dependent.

White spot syndrome virus (WSSV) is one of the most devastating pathogens of shrimps and other crustaceans. In a previous study, we demonstrated that the major envelope protein VP28 of WSSV was involved in the attachment and penetration into shrimp cells. Here we showed that heat-shock cognate protein 70 (Hsc70) of shrimp was a binding partner of VP28 during virus infection. Hsc70 expression was enhanced by WSSV infection at the early stage and peaked at 12h post-infection and decreased drastically at the late stage. In shrimp haemocytes, both VP28 and Hsc70 proteins localized in the cytoplasm and their association was specific, ATP-dependent and Hsc70 concentration dependent. Moreover, direct VP28-Hsc70 association required both ATPase domain and peptide binding domain of Hsc70. The data obtained will help elucidate the role of VP28 protein in the process of virus infection.

Retroperitoneoscopic Anderson–Hynes dismembered pyeloplasty in infants and children: a 60-case report

PurposeTo present a new technique of retroperitoneoscopic Anderson–Hynes dismembered pyeloplasty (AHDP) in infants and children with ureteropelvic junction obstruction (UPJO) based on our clinical experience.MethodsFrom March 2003 to February 2007, retroperitoneoscopic AHDP was performed in 60 (44 boys and 16 girls) UPJO infants and children with a three-port lateral retroperitoneal approach. The retroperitoneal space was entered via a 1-cm longitudinal incision beneath the 12th rib and further developed by a glove balloon. Video-retroperitoneoscopy was undertaken with a 5-mm laparoscope between the mid axillary line and 1xa0cm away from the superior border of iliac crest. Dismembered pyeloplasty was carried out with the Anderson–Hynes anastomosis where 5-0 or 6-0 vicryl sutures were over a double-J ureteric stent. Anastomosis was completed with freehand intracorporeal suture techniques. Follow-up studies were conducted by intravenous urography and renal ultrasonography.ResultsAmong the 60 patients (62 sides) with retroperitoneoscopic AHDP, only the first two cases were converted to open surgery due to difficulties in developing the retroperitoneal space, and the remaining cases succeeded. The average operative time was 70xa0±xa012.6xa0min (ranging from 55 to 130xa0min), the average estimated blood loss was 10xa0±xa02.2xa0ml (ranging from 5 to 20xa0ml), and the average postoperative hospital stay was 7xa0±xa01.3xa0days (ranging from 3 to 15xa0days). Aberrant artery vessel was intraoperatively observed in seven patients. Postoperative urinary leakage occurred in two patients, but spontaneously disappeared on the 6th and 11th days after the surgery, respectively; and one of them underwent open surgery for recurrent UPJO 8xa0months later. During an average follow-up period of 24xa0months, we performed radiographic assessment by intravenous urography and found that all the cases showed good results except the patient who underwent open surgery later.ConclusionsOur experience with retroperitoneoscopic AHDP demonstrates that this technique is safe, effective and time saving for treating UPJO in infants and children.

The retroperitoneal laparoscopic Hellström technique for pelvi-ureteric junction obstruction from a crossing vessel

Associate Editor

Laparoscopic Adrenalectomy for Beginners Without Open Counterpart Experience: Initial Results Under Staged Training

OBJECTIVESnTo develop a staged laparoscopic training program for beginners to perform laparoscopic adrenalectomy (LA) and to determine its safety and feasibility.nnnMETHODSnFrom January 2002 to October 2007, 5 beginners (postgraduate years 1-5) without previous experience in open adrenalectomy were selected randomly to receive the staged laparoscopic training, including box-trainer, animal model, and mentor-initiated clinical training. During the clinical training, the trainees acted as the camera holder first, and then selectively performed simple operations, such as laparoscopic renal cyst unroofing. Finally, they performed 30 LAs independently under the mentors supervision using the technique of anatomic retroperitoneoscopic adrenalectomy. The clinical data of the 30 LAs performed by each the trainees (150 LAs total) were collected and compared with the data from the initial 30 LAs of the mentor.nnnRESULTSnAll LAs were completed successfully. No procedure required conversion to open surgery. The median operative time of the trainees was 82.3 minutes (range 59-133), which was obviously shorten than the mentors (median operative time 131.5 minutes, range 73-230, P < .001). The learning curve among the trainees was shorter compared with that of the mentor. No major complications were observed. The minor intraoperative and postoperative complication rate for the trainees was 0.67% and 6.7%, respectively, not significantly different from those of the mentor (0% and 3.3%, respectively; both P > .05). All complications developing in patients treated by the trainees required only conservative therapy.nnnCONCLUSIONSnIt was safe and feasible for beginners without previous open counterpart experience to perform LA using staged training.

Relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene in bladder tumor

SummaryTo investigate the relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene, RASSF1A protein expression was measured by Western blotting in 10 specimens of normal bladder tissues and 23 specimens of bladder transitional cell carcinoma (BTCC). The promoter methylation in BTCC and normal bladder tissues was detected by methylation-specific PCR (MSP). The results showed that the expression level of RASSF1A protein was significantly lower in BTCC tissues than that in normal bladder tissues. However, it was not correlated with its clinical stages and pathological grades. The frequency of promoter methylation of RASSF1A gene was higher in BTCC tissues than that in normal bladder tissues. In 14 patients with the aberrant promoter methylation, 13 showed loss or low expression of RASSF1A protein. It is concluded that RASSF1A gene promoter methylation may contribute to the low level or loss of RASSF1A protein expression, the inactivation of RASSF1A gene and the genesis of BTCC. But, it may bear no correlation with its clinical stages and pathological grades.To investigate the relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene, RASSF1A protein expression was measured by Western blotting in 10 specimens of normal bladder tissues and 23 specimens of bladder transitional cell carcinoma (BTCC). The promoter methylation in BTCC and normal bladder tissues was detected by methylation-specific PCR (MSP). The results showed that the expression level of RASSF1A protein was significantly lower in BTCC tissues than that in normal bladder tissues. However, it was not correlated with its clinical stages and pathological grades. The frequency of promoter methylation of RASSF1A gene was higher in BTCC tissues than that in normal bladder tissues. In 14 patients with the aberrant promoter methylation, 13 showed loss or low expression of RASSF1A protein. It is concluded that RASSF1A gene promoter methylation may contribute to the low level or loss of RASSF1A protein expression, the inactivation of RASSF1A gene and the genesis of BTCC. But, it may bear no correlation with its clinical stages and pathological grades.

Polymorphisms in CYP11B2 and CYP11B1 genes associated with primary hyperaldosteronism.

Several frequent polymorphisms in the CYP11B2 gene are suggested to be associated with essential hypertension and aldosterone secretion. In this study, we investigated the association of polymorphisms in CYP11B2 and CYP11B1 genes with the risk of primary hyperaldosteronism (PH). Three polymorphisms in the CYP11B2 gene (intron 2 conversion, rs1799998 and rs4539) and two polymorphisms in the CYP11B1 gene (rs6410 and rs6387) were analyzed in patients with PH and in the normal population. The rs6410 allelic frequencies in patients with aldosterone-producing adenoma (APA) and idiopathic hyperaldosteronism (IHA) were significantly different from those in controls at P=1.09 × 10−5 and 0.015, respectively. There was a relative excess of AA homozygotes and AG heterozygotes of the rs6410 allele in the APA group as compared with the control group (P=2.19 × 10−4). There were significantly different genotypes, AA and AG, of the rs6410 allele between the patients with IHA and the controls only after adjustments for age, gender and body mass index (odds ratio (OR)=4.06, 95% confidence interval (CI) 1.31–12.66; OR=2.41, 95% CI 1.02–5.72). One susceptible haplotype, AAAWT, was identified to be significantly associated with APA (OR=1.44, 95% CI 1.19–1.76), and 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, one protective haplotype, GGAWT, showed a significant difference between the patients with APA and controls (OR=0.73, 95% CI 0.55–0.97). Several haplotypes were associated with ARR in both the controls and cases. Our data demonstrated that there was a significant association between polymorphisms in the CYP11B2 and CYP11B1 genes and a genetic predisposition to PH. The association with IHA seemed closer compared with APA.

DNA polymorphisms in exon 1 and promoter of the CDH1 gene and relevant risk of transitional cell carcinoma of the urinary bladder

To investigate the association between DNA polymorphisms, including single‐nucleotide polymorphisms (SNPs) and insertion/deletion polymorphisms, in exon 1 and promoter of the CDH1 gene, and the risk of transitional cell carcinoma (TCC) of the urinary bladder (TCCB).

MicroRNA-30a as a prognostic factor in urothelial carcinoma of bladder inhibits cellular malignancy by antagonising Notch1.

To explore the relation between microRNA‐30a (miR‐30a) and Notch1, and to evaluate the potential prognostic role of miR‐30a in invasive urothelial carcinoma of the bladder (UCB).

Optimization of Douchi fibrinolytic enzyme production by statistical experimental methods

Thrombus disease, one of the common cardiovascular diseases, has attracted worldwide attention for its rising mortality and morbidity. Due to the distinct shortages of current fibrinolytic drugs, new fibrinolytic agents warrant investigation. In this study, 8 fibrinolytic enzyme-producing strains were isolated from Douchi—a traditional Chinese food, and strain XY-1 which produced the largest amount of the enzyme was chosen for the following experiments. The enzyme produced by strain XY-1 was named Douchi fibrinolytic enzyme (DFE). We optimized the liquid culture medium of strain XY-1 for enzyme production using Plackett-Burman and Box-Behnken design. The predicted maximal DFE yield was 19.78 FU/mL with 11.4 g/L peptone, 0.5 g/L magnesium sulfate and 1 g/L sodium chloride. However, we acquired maximal production of 21.33 FU/mL in actual experiments, equal to 107.84% of the theoretical value, and the yield had been increased by 79.55% as compared to the yield of un-optimized culture. It was demonstrated that the combined use of Plackett-Burman design and response surface methodology in fermentation optimization can effectively and rapidly increase DFE production.SummaryThrombus disease, one of the common cardiovascular diseases, has attracted worldwide attention for its rising mortality and morbidity. Due to the distinct shortages of current fibrinolytic drugs, new fibrinolytic agents warrant investigation. In this study, 8 fibrinolytic enzyme-producing strains were isolated from Douchi—a traditional Chinese food, and strain XY-1 which produced the largest amount of the enzyme was chosen for the following experiments. The enzyme produced by strain XY-1 was named Douchi fibrinolytic enzyme (DFE). We optimized the liquid culture medium of strain XY-1 for enzyme production using Plackett-Burman and Box-Behnken design. The predicted maximal DFE yield was 19.78 FU/mL with 11.4 g/L peptone, 0.5 g/L magnesium sulfate and 1 g/L sodium chloride. However, we acquired maximal production of 21.33 FU/mL in actual experiments, equal to 107.84% of the theoretical value, and the yield had been increased by 79.55% as compared to the yield of un-optimized culture. It was demonstrated that the combined use of Plackett-Burman design and response surface methodology in fermentation optimization can effectively and rapidly increase DFE production.