Long­term treatment with salicylate enables NMDA receptors and impairs AMPA receptors in C57BL/6J mice inner hair cell ribbon synapse.

Salicylate is widely used to produce animal models of tinnitus in mice and/or rats. The side effects on auditory function, including hearing loss and tinnitus, are considered the results of the auditory nerve dysfunction. A recent study indicated that chronic treatment with salicylate for several weeks reduces compressed action potential amplitude, which is contradictory to the studies reporting excessive activation of N­methyl­D­aspartate receptors (NMDAR) in tinnitus­induced animals. The specific aims of the experiment were to detect the effect of salicylate on the inner hair cells (IHCs), ribbon synapse, as well as the association between the hearing threshold and the number of mismatched ribbon synapses. In the present study, mice were injected intraperitoneally with a low dose of salicylate (200 mg/kg) for 14 days. The auditory brainstem response and otoacoustic emission were measured to assess auditory function of the mice. The postsynaptic regions of IHC were identified with two types of immunostaining targets: Postsynaptic density protein 95 and Glu2/3. The number of spheres was counted and the synapses were reconstructed in 3­dimensional images. Increases in distortion product otoacoustic emissions amplitudes of the salicylate group were detected, however, an elevation in the hearing threshold was also observed. A mismatch between pre­and post­ribbon synapses was observed. In addition, the cochlear components, including the numbers of outer hair cells and IHCs, were unlikely to be affected by salicylate. IHC ribbon synapses were more susceptible to salicylate stimuli. Furthermore, mismatch of pre­ and post­ribbon synapses may indicate a competitive inhibition between NMDAR and α-amino­3­hydroxy-5-methyl-4-isoxa-zole-propionate receptors and dysfunction of ribbon synapses.

Prospective validation of DACH2 as a novel biomarker for prediction of metastasis and prognosis in muscle-invasive urothelial carcinoma of the bladder.

Metastasis is the main cause of death from muscle-invasive urothelial carcinoma of the bladder (UCB), and the metastatic potential of tumors is often unpredictable. The role of Dachshund homolog 2 gene (DACH2) in tumorigenesis remains unexplored. We aimed to investigate whether DACH2 can be used as a biomarker to predict metastasis and prognosis of muscle-invasive UCB in a sequential training and validation fashion. For the training set (n = 40), compared with UCB patients without lymph node (LN) metastasis, both DACH2 protein and mRNA expression were greatly increased in case-matched patients with LN metastasis. For the independent validation set (n = 243), patients with primary UCB that did not express DACH2 had a longer metastasis-free survival (MFS) and overall survival (OS) than did those with tumors expressing DACH2 (5-year MFS: 88% [95% CI 80-96] versus 19% [95% CI 7-31], p < 0.001; 5-year OS: 93% [95% CI 87-99] versus 37% [95% CI 23-51], p < 0.001). Multivariable analysis of DACH2 status showed hazard ratios of 7.34 (95% CI 3.15-11.87, p < 0.001) for MFS and 3.96 (95% CI 2.04-7.16, p < 0.001) for OS which were much higher than hazard ratios associated with other independent risk factors. Collectively, DACH2 is an independent prognostic marker that can be used at initial diagnosis of UCB to identify patients who have a high potential to develop metastasis.

Loupe-assisted versus microscopic varicocelectomy: is there an intraoperative anatomic difference?

The aim of this study was to compare the intraoperative difference in anatomic details between loupe-assisted and microscopic varicocelectomy within the same spermatic cord. Between April 2011 and August 2011, 26 men with 33 sides containing grade 2-3 varicocele were enrolled in this study. First, one surgeon performed the open inguinal varicocelectomy under × 3.5 loupe magnification. The presumed vascular channels and lymphatics were isolated and marked without ligation. Another surgeon then microsurgically dissected and checked the same spermatic cord using an operating microscope to judge the results in terms of the ligation of the internal spermatic veins and the preservation of the arteries and lymphatics. There were significant differences in the average number of internal spermatic arteries (1.51 vs 0.97), internal spermatic veins (5.70 vs 4.39) and lymphatics (3.52 vs 1.61) between the microscope and loupe-assisted procedures (P < 0.001, P < 0.001, P < 0.001, respectively). Meanwhile, in varicocele repair with loupe magnification, an average of 1.30 ± 1.07 (43/33) internal spermatic veins per side were missed, among the overlooked veins, 1.12 ± 0.93 (37/33) were adhered to the preserved testicular artery, as well as 0.55 ± 0.79 lymphatics and 0.36 ± 0.55 arteries that were to be ligated. In conclusion, microscopic varicocelectomy could preserve more internal spermatic arteries and lymphatics and could ligate more veins than the loupe-assisted procedure. To some degree, loupe magnification is inadequate for the reliable identification and dissection of the tiny vessels of the spermatic cord, as most of the overlooked veins were adhered to the preserved testicular artery.

Microsurgical transverse 2-suture intussusception vasoepididymostomy: effectiveness and rationality.

BACKGROUND: 2-Suture longitudinal vasoepididymostomy shows superiority to transverse technique in an animal study; to date, this has not been consistently confirmed in human body. In the present study, we evaluated the effectiveness of 2-suture transverse intussusception vasoepididymostomy and compared the rationality between transverse and longitudinal techniques. METHODS: From May 2007 to December 2008, we performed 2-suture transverse vasoepididymostomy in 19 consecutive patients, as described by Marmar with modification. Between March 2009 and January 2010, the internal diameter of the vas lumen and the outer diameter of the epididymal tube were measured using microruler (21 patients and 37 sides). RESULTS: Three patients lost to follow-up. At the first follow-up period (ranged from 10 to 24 months), the patency rate was 56.3% (9/16) and the natural pregnancy rate was 25% (4/16). At the second follow-up period (ranged from 46 to 63 months), the patency rate was 68.8% (11/16), the natural pregnancy rate was 37.5% (6/16), respectively, and the take-home baby rate was 31.3% (5/16). The diameter of the vas lumen and the outer diameter of the epididymal tubule were (0.512 ± 0.046) mm and (0.572 ± 0.051) mm (P < 0.001), respectively. CONCLUSION: Transverse 2-suture intussusception vasoepididymostomy is still an effective technique in treating obstructive azoospermia.

Inhibition of extracellular signal-regulated kinase activity by sorafenib increases sensitivity to DNR in K562 cells.

The mitogen-activated protein kinase (MAPK) pathway has a protective function on the management of hematologic malignancies. The aim of this study was to assess whether the induction of MAPK-mediated effects contributes to the therapeutic value of combination sorafenib and daunorubicin (DNR) treatment. Herein, we found that DNR increased phosphorylation of extracellular signal-regulated kinases (ERK1/2) in K562 cells. ERK1/2 activity was blocked by either the mitogen-induced extracellular kinase (MEK) inhibitor U0126 or a multi-kinase inhibitor sorafenib. Of note, sorafenib sensitized K562 to DNR by inhibiting proliferation and inducing apoptosis in a dose-dependent manner which was through blocking the RAF/MEK/ERK pathway. Moreover, K562 cells transfected with a constitutively active MEK2DD plasmid showed increasing IC50 values following DNR treatment compared with control cells. Combination of DNR with MEK inhibitor U0126 synergistically inhibited K562 cell growth. In conclusion, our results indicated that sorafenib sensitized K562 cells to DNR-induced cytotoxicity by downregulating p-ERK1/2 expression. DNR in combination with sorafenib may represent a new and potential therapeutic strategy in treating acute leukemia with high p-ERK1/2 levels.

Macroscopic and microsurgical varicocelectomy: what's the intraoperative difference?

PURPOSE: Many authors reported that microsurgical varicocelectomy was among the best treatment modalities for varicocele. However, the difference in intraoperative anatomic detail between macroscopic and microsurgical varicocele repair in the same spermatic cord has not been critically discussed. METHODS: Between August 2010 and February 2011, 32 men with 42 sides' grade 2-3 varicocele were enrolled in this study. One surgeon firstly mimicked the modified open varicocelectomy by identifying, isolating, and marking the presumed internal spermatic veins, lymphatics, and arteries. Another surgeon then checked the same spermatic cord using operating microscope to investigate the number of missed veins, to be ligated lymphatics and arteries in the "imitative" open varicocelectomy. RESULTS: There were significant differences in the average number of internal spermatic arteries (1.67 vs. 0.91), internal spermatic veins (6.45 vs. 4.31), and lymphatics (2.93 vs. 1.17) between microscopic and macroscopic procedure (P < 0.001, P < 0.001, P < 0.001, respectively). Meanwhile, an average of 2.14 ± 1.26 internal spermatic veins was missed; among them, 1.63 ± 1.32 internal spermatic veins adherent to the preserved testicular artery were overlooked. The number of 0.69 ± 0.84 lymphatics and 0.74 ± 0.74 arteries were to be ligated in "macroscopic varicocelectomy." A number of 1.07 ± 1.11 lymphatics were neither identified nor ligated. In addition, in 2 cases, the vasal vessels of the vas deferens were to be ligated at macroscopic procedure. CONCLUSIONS: Microsurgical varicocelectomy could preserve more internal spermatic arteries and lymphatic and ligate more veins which may interpret the superiority of microsurgical varicocele repair.

[Sorafenib induces apoptosis of U937 cells via inhibiting WNT signal pathway].

This study was aimed to investigate the effect of multikinase inhibitor sorafenib on the proliferation and apoptosis of U937 cells and its possible mechanism. U937 cells were treated with different concentrations of sorafenib for 48 hours. Cell viability was determined by Cell Counting Kit-8; cell apoptosis and cell ratio in cell cycle were detected by flow cytometry with Annexin V/PI staining and PI staining respectively; expressions of GSK-3ß, ß-catenin and cyclin-D1 were assayed by Western blot. The results showed that the proliferation of U937 cells was inhibited by sorafenib in a dose-dependent manner (p < 0.05). Sorafenib induced cell apoptosis and cell cycle G(1)/G(0) arrest. Compared with results of Western blot before treatment, expression of inactivated GSK-3ß, ß-catenin and Cyclin-D1 down-regulated in a dose-dependent manner after treatment with sorafenib, this same changes were observed after up-regulation of inactivated GSK-3ß by LiCl (p < 0.05). It is concluded that sorafenib inhibits the proliferation of U937 cells and induces cell apoptosis through reducing negative regulation of WNT signal pathway on inactivated GSK-3ß and down-regulating ß-catenin and cyclin-D1 level, which result in U937 cell cycle G(1)/G(0) arrest.

[Effect of sorafenib combined with daunorubicin on K562 cell line].

The aim of this study was to investigate the effect of sorafenib combined with daunorubicin on leukemic k562 cell line. The inhibitory effect of sorafenib alone and its combination with daunorubicin on K562 cell proliferation was detected by MTT method; the synergistic effect was measured by CDI (coefficient of drug interaction); the apoptosis of K562 cells was observed by flow cytometry with Hoechst 33258 staining. The results showed that the sorafenib alone or its combination with daunorubicin could significantly inhibit K562 cell proliferation and the combination of both drugs displayed synergistic effect on K562 cells, meanwhile the apoptotic cells increased. It is concluded that the combination of sorafenib and daunorubicin has a obviously synergistic inhibitory effect on leukemic cell line K562.

Application of a temporary ureter clamp for retroperitoneal laparoscopic ureterolithotomy.

INTRODUCTION: Retroperitoneal laparoscopic ureterolithotomy is a new option to treat upper and middle ureter calculi in selected patients. However, migration of the stone and the flow of urine will influence the success. METHODS: We have developed a new and practical method using a clamp proximally above the stone to fix its position and prevent urine flow. Twenty patients had undergone retroperitoneal laparoscopic ureterolithotomy using this method. A bulldog artery clamp was used as the temporary clamp and was placed at the proximal part of the ureter during the procedures of incision, intubation and suture. RESULTS: The average age of the patients was 42.5 years (range: 26-73) and the average stone size was 13.7 mm (range: 10-28). The average operating time was 38.2 min (range: 30-85). All target stones were successfully extracted without major complications. The average time of post-operation drain removal was 1.5 days. No case of prolonged urine leakage or ureter stricture was recorded. CONCLUSIONS: We conclude that using a temporary ureter clamp is a feasible and practical method to fix the stone and minimize the difficulty of retroperitoneal laparoscopic ureterolithotomy.

Cyclooxygenase-2 expression is associated with vascular endothelial growth factor-C and lymph node metastases in human prostate cancer.

BACKGROUND AND AIMS: Recent observations suggest an implication of cyclooxygenase-2 (COX-2) in tumor lymphangiogenesis through an upregulation of vascular endothelial growth factor-C (VEGF-C) expression. However, it is unclear whether COX-2 is also associated with VEGF-C expression, tumor lymphangiogenesis and lymph node metastasis in human prostate cancer. METHODS: COX-2 and VEGF-C expression were examined in tumor tissues from 58 prostate cancer patients using immunohistochemical staining. We also analyzed the association of COX-2 and VEGF-C expression with tumor lymphangiogenesis quantified as lymphatic vessel density (LVD), lymph node metastasis, and patients' biochemical progression-free survival (b-PFS). RESULTS: High expression of either COX-2 or VEGF-C was correlated with tumor lymphangiogenesis and lymph node metastasis, as well as poor b-PFS. Moreover, a strong correlation was found between expression of COX-2 and VEGF-C (r = 0.631, p <0.001). CONCLUSIONS: COX-2 is positively associated with VEGF-C expression, tumor lymphangiogenesis and lymphatic metastasis in prostate cancer. These findings suggest that COX-2 may play a pivotal role in lymphangiogenesis and lymph node metastasis of prostate cancer via the regulation of VEGF-C expression.

Toll-like receptor 9 agonists promote IL-8 and TGF-beta1 production via activation of nuclear factor kappaB in PC-3 cells.

Chronic infection and resulting inflammation promote tumor development and progression, and Toll-like receptors (TLRs) may play an important role in this process. The aim of this study was to determine whether CpG oligonucleotides (CpG-ODN), which are Toll-like receptor 9 (TLR9) agonists, can promote inflammatory cytokines release from the prostate cancer PC-3 cells through activation of nuclear factor-kappaB (NF-kappaB). Flow cytometry, semiquantitative real-time reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent assay, and immunofluorescence analysis were used to detect the transforming growth factor-beta1 (TGF-beta1) and interleukin-8 (IL-8) release and NF-kappaB activation in PC-3 cells after CpG-ODN stimulation. CpG-ODN promoted the expression and secretion of immunosuppressive cytokines TGF-beta1 and IL-8 from PC-3 cells. In addition, after CpG-ODN stimulation, NF-kappaB nuclear translocation was also observed in PC-3 cells, contributing to CpG-induced upregulation of IL-8 and TGF-beta1. Thus, TLR9 agonists may promote IL-8 and TGF-beta1 production in human prostate cancer cells through NF-kappaB activation.