Objective: To construct DNA vaccines expressing prostate-specific membrane antigen (PSMA) and/or granulocyte-macrophage colony-stimulating factor (GM-CSF) and to determine their immunoactivity. Methods: Recombinant plasmids pIRES-PS-MA-mGM-CSF, pIRES-PSMA, and pIRES-mGM-CSF were constructed with DNA vaccine vector pIRES. After identified by endonuclease digestion, the above 3 plasmids and blank pIRES vector were used to immunize C56BL/6 mice (n=15). LDH release assay was used to exam the cytotoxicity of cytolytic T lymphocytes in each group. Results: We successfully constructed the above mentioned recombinant plasmids. Mice in pIRES-PSMA-mGM-CSF immunized group had the highest specific cytotoxicity, followed by pIRES-PS-MA and pIRES-mGM-CSF immunized groups. The blank pIRES group had the lowest cytotoxicity (P<0.05). The cytotoxicity was the highest in all 4 groups at an effector/target ratio of 10/1. Conclusion: The bicistronic DNA vaccine expressing PSMA and mGM-CSF may have a promising therapeutic value in gene therapy of prostate cancer.

Objective: To assess the efficacy and safety of the ureteroscopic lithotripsy in treatment of ureteral calculi in children. Methods, From March 2008 to May 2009, thirteen pediatric patients with ureteral stones were admitted to our department. There were six males and seven females, with an age range of 23 months to 11 years old. Six patients with stones located at the lower segments of the ureter, four at the middle segments, two at the upper segments, and one at pyelo-ureteral junction and lower ureter. The average diameter of the stones was (1.1 ± 0.47) cm (range 0.6 - 2.0 cm). The patients were placed in a lithotomy position; under general or venous anesthesia, ureteroscopy was performed using a semirigid ureteroscope (WOLF Fr 7.5/6), and the ureteral orifice was dilated to 10-12 Fr. Then holmium laser (1.5 J, 10 Hz) or pneumatic lithotripter was employed to fragment the stones (diameter of fragment less than 3 mm). Fr4. 7 double-J stent was placed after operation and was removed 4 weeks later. Results: Eleven of the 13 patient underwent successful lithotripsy, with the first time successful rate being 84.6%. The mean lithotripsy time was 8 min and the mean operation time was 21 min. There was slight bleeding during operation, but with no noticeable perforation of ureter or post operation fever, etc. Post-operation B ultrasound examination and X ray image revealed no residual stones with diameter > 3 mm in the 11 cases. Partial fragmentation was achieved in one patient with stone at upper ureter segment and one patient with stone at pyelo-ureteral junction and lower ureter, and the stones were excreted after extracorporreal shock wave lithotripy. Conclusion: Ureteroscopic lithotripsy is safe and effective in treatment of children with ureteral stones, which might become one of the first line treatment for children.

Objective To evaluate the clinical value of the PolyScope™ endoscope system (F8 modular flexible ureteroscope) in treatment of upper urinary calculi less than 2 cm in diameter. Methods A total of 117 patients with upper urinary calculi, who were hospitalized in Xinhua Hospital from Nov. 2010 to Aug. 2012, were recruited in this study. The patients included 71 males and 46 females, aging from 20-86 (average age:49.7±13.9). Ninety-six patients had single calculi and the rest 21 had multiple calculi; 61 patients had left calculi and 56 had right calculi. The mean diameter of calculi was (1.19±0.34) cm. Twenty-two patients had been indwelled with D-J tube in advance under ureteroscope for 2-8 weeks, including 13 with ureteral stenosis, 7 with urinary tract infection and 2 with renal insufficiency. General or spinal anaesthesia were used and the patients were at cutting stone position. F8/9.8 rigid ureteroscope was inserted to the ureter for dilating the ureter, and the Holmium laser lithotripsy was performed for upper ureteral stone in situ. After dilating the ureter, the guide wire was inserted under the direct vision of F8/9.8 rigid ureteroscope, and then the F12/14 flexible ureteral access sheath was placed along the guide wire. Finally, further examination and Holmium laser lithotripsy were performed by F8 "PolyScope" modular flexible ureteroscope. The power of the laser was set at 1.0 J and the frequency was 10-20 Hz. F6 D-J tube was regularly indwelled for 4 weeks after operation and Forley urethral tube was indwelled for 1 to 7 days. KUB was performed 1 day after operation to detect the result of lithotropsy and the position of D-J tube. KUB was performed again 2-4 weeks after operation in some patients. Extracorporeal shock wave lithotripsy (ESWL) should be performed if the diameter of residual stones was larger than 6 mm. Results Lithotripsy was successfully performed in 103 patients, with the operation time being 25-85 min (mean time: [46±14] min) and with less bleeding. Four patients developed high fever after operation and were cured after anti-inflammatory treatment. There was no ureteral perforation or septicaemia. The single stone-free rate of single-pass lithotripsy was 88% (103/117); when combined with ESWL, the total stone-free rate was 95.7% (112/117).Conclusion F8 modular flexible ureteroscope is safe, convenient, and effective for the lithotripsy of the upper urinary calculi.

Objective To discuss the reasons and management of renal vein injury during percutaneous nephrostolithotomy (PCNL). Methods From 2007 to 2008 renal vein injury was caused in two patients by malposition of nephrostomic catheter in our hospital. The two patients were both males, aged 61 years old and 41 years old. They underwent PCNL due to left kidney stones. The percutaneous nephrostomy (PCN) catheters were malpositioned into the left kidney vein and vena cava, which was confirmed by prograde radiography and spiral CT scan after operation. The PCN catheter in the 41 years old patient was pulled back to renal collecting system on the 7th day, and was extracted on the 10th day under X-ray monitoring. The PCN catheter in the 61 years old patient was pulled back to the renal collecting system on the 14th day and wasstable extracted on the 18th day under X-ray monitoring. Results The bleeding was controlled and the hemodynamic status in the two cases after removal of PCN catheters. There was no renal arteriovenous fistula bleeding, surgical intervention, kidney infections or further damage of kidney function. Conclusion Renal vein injury during the PCNL can be managed by clamping the PCN catheter and gradual withdrawal. The method is safe and reliable, and it can avoid surgical intervention.

Objective To investigate the association of genetic polymorphisms of intercellular adhesion molecule 1 (ICAM-1) with diabetic peripheral neuropathy (DPN). Methods A total of 607 type 2 diabetes patients from the Affiliated Hospital of Weifang Medical University were enrolled in this study between June 2013 and December 2014. Rs5498 (A/G K469E) and rs1799969 (G/A R241G) in the ICAM-1 gene were genotyped by using TaqMan allelic discrimination in 295 patients with DPN and 312 subjects without DPN. The distribution of these two SNPs and the genetic influence of ICAM-1 gene polymorphisms on the development of DPN were conducted. Results Genotype distributions of both SNPs were coincided with Hardy-Weinberg equilibrium in the two groups. SNP rs1799969 (G/A R241G) in the ICAM-1 gene showed a high GG genotypic frequency at 96.8%(non DPN) and 99.0%(DPN) respectively. SNP rs5498 (A/G K469E) represented AA and AG genotypes. The values were AA 48.7%/AG 39.4%in non DPN group and AA 51.5%/AG 41.7%in DPN group. There were no significant differences in genotypic distributions and allele frequencies of SNPs rs1799969 (G/A R241G) and rs5498 (A/G K469E) between the patients with DPN group and patients without DPN group (P>0.05). The dominant(AA+AG)/GG and additive (GG/AA) models of rs5498 (A/G K469E) were associated with higher risk of DPN (ORadjusted=1.585, 1.575 respectively, P<0.05). To carry A allele was related to the susceptibility of DPN. There was no such association in genetic models of rs1799969 (G/A R241G) and DPN pathogenesis. Conclusion The present study provides evidence that SNP rs5498 E469K (A/G) in the ICAM-1 gene is associated with susceptibility of DPN, and the carrying A allele appears to be a risk of DPN.

OBJECTIVESTo evaluate modified cystectomy with preservation of erectile and ejaculatory functions in men with nonmalignant bladder disease.

METHODSSeven cases with average age of 27 years presented with bladder disease necessitating cystectomy, including 2 cases of tuberculous contractile bladder, 1 case of extensive polypoid cystitis glandularis, 4 cases of late stage of neurogenic bladder. All patients wished to maintain erectile and ejaculatory functions after the operation. We performed a modified simple cystectomy with preservation of the vasa deferens, seminal vesicles, prostate and neurovascular bundles, as well as construction of an Indiana pouch or ileal neobladder.

RESULTSAverage operative time was 5 h 45 min without perioperative complications in this group. Follow-up ranged from 9 to 60 months. Erectile and ejaculatory functions were normal in all cases. All patients remained completely continent and no dysuria in neobladder, and there was no difficulty in inserting catheter to empty pouch. Upper urinary tract was in good condition 3 and 24 months after operation.

CONCLUSIONSModified cystectomy with preservation of the vasa deferens, seminal vesicles, prostate and neurovascular bundles is an effective and reliable option for the patients who wish to maintain their fertility and erectile function after surgery.

Adult ; Cystectomy ; methods ; Ejaculation ; physiology ; Humans ; Male ; Middle Aged ; Penile Erection ; physiology ; Urinary Bladder Diseases ; surgery

Objective:To evaluate the efficacy and toxicity of single-agent bendamustine in patients with indolent B-cell non-Hodgkin's lymphoma (NHL) refractory to rituximab. Methods:Between April 2010 and April 2013, 100 patients with rituximab-refrac-tory indolent B-cell NHL from 8 institutions were enrolled. Bendamustine was administered at 120 mg/m2 on days 1 and 2 every 21 days for 6-8 cycles. The primary endpoint was the overall response rate (ORR). The secondary endpoints included disease control rate (DCR), progression-free survival (PFS), overall survival (OS), and safety. Results:One hundred patients with a median age of 56 (rang-ing from 28 to 74) years were recruited in this clinical study. The total number of chemotherapy was 447 cycles, and the median number was 4 cycles. Ninety-three patients could be evaluated for efficacy. Fifteen patients (16.1%) had complete remission (CR), 52 (55.9%) had partial remission (PR), 22 (23.7%) had stable disease (SD), and 4 (4.3%) had progression disease (PD). The ORR and DCR were 72%and 95.7%, respectively. After a median follow-up of 26.6 months (ranging from 2 to 48.4 months), 59 patients (63.4%) had PD.The median PFS was 8.53 (95%CI:6.518-10.542) months, and PFS rate for 1 year was (40.6±5.3)%. Forty-eight patients (48%) had 3/4 grade adverse events, including leucopenia (26%), neutropenia (24%), and anemia (11%). Conclusion:Single-agent bendamustine produced a high rate of objective responses in patients with rituximab-refractory indolent B-cell NHL and could be one of the new op-tions for second-line treatment of these patients. The most common adverse event is hematologic toxicity.

Congenital heart disease is one of the most common birth defects, and its structural changes often require surgical or non-surgical intervention after delivery. Some scholars believe that because early primary intrauterine heart abnormalities may hinder the normal development of the heart and cause irreversible secondary structural changes, the current treatment of congenital heart disease focuses on prenatal diagnosis and fetal intervention. Since the first successful fetal surgery in 1984, and with the development and progress of diagnostic techniques such as fetal cardiac ultrasound, the treatment of congenital heart disease has gradually become the focus of research. This article will focus on the intrauterine cardiac intervention of a specific type of congenital heart disease, fetal cardiac surgery guided by fetal scope, the impact of cardiopulmonary bypass on the fetus or the mother, intrauterine cardiac surgery and targeted therapy of fetal congenital heart disease are summarized in this article.

Objective To explore the expression level of CD36 in peripheral blood mononuclear cells(PBMCs) from patients with crytococcal meningitis and its clinical significance.Methods The experimental group was the peripheral blood samples from 36 patients diagnosed as crytococcal meningitis in Changhai Hospital and Changzheng Hospital in Shanghai from September 2013 to February 2017,while the control group was the peripheral blood samples from 36 health individuals examined at the same time.PBMCs were separated by density gradient centrifugation method,the relative mRNA expression of CD36,TLR4 in PBMCs was tested by qRT-PCR,the expression level of cytokines such as TNF-α in plasma was tested by ELISA,the comparison of the examined values of experimental and control groups performed by two independent samples' t test,Pearson correlation analysis was performed on the relative mRNA expression of CD36 in PBMCs and cytokines such as TNF-α in experimental group.Results The relative mRNA expression of CD36 in PBMCs of experimental group was 2.01 ± 0.63,the control was 1.49 ± 0.47,and there was statistical difference (t =3.969,P =0.000 2).Pearson analysis results showed that in experimental group CD36 was positively related with the protein expression level of IFN-γ in plasma (r=0.384,P＜0.05) and negatively related with TGF-β(r=-0.487,P＜0.005).Conclusion CD36 might involve in the immunopathology process of crytococcal meningitic by influencing the expression of cytokines such as IFN-γ,TGF-β,and was the potential target for monitoring and treating this disease.

Objective To study the expression of CD69 in exosomes existed in serum from patients with autoimmune pancrea titis (AIP) and primarily discuss its affection on the pathology of disease progression.Methods Serum samples from 35 cases diagnosed as AIP and 35 healthy individuals examined at the same time were collected during October 2012 to December 2016 in Changhai Hospital,they were defined as experimental and control groups,separately.The cytokine levels in serum were measured by ELISA.The expression of surfaced molecules in exosomes existed in serum was tested by magnetic bead conjunct antibody method in flow cytometry.The comparison between the two groups' measurement data was measured by two independent samples' t test,correlation between two variables was showed by Pearson coefficient.Results The flow cy tometry analysis showed that the expression level of CD69 in serum exosomes in experiment and control groups were (85.76 ±19.45 vs 17.01±5.89)％,with statistical difference(t=20.01,P＜0.0001).CD69+ exosomes in experiment group were positively related with serum IL-4,IFN-γ and TGF-β (r 0.456,0.678,0.548,all P＜0.05),while CD69 exosomes were negatively related with serum IL-4,IFN-γ,IL-10 and TGF-β (r=0.589,-0.399,-0.784,-0.657,all P＜0.05).Conclusion The expression of CD69 in exosomes might participate in AIP progression through influencing the function of CD4 +T cell,and it was the potential examined biomarker and therapeutic target.