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Purpose: To explore the mechanisms underlying clear-cell renal cell carcinoma (ccRCC) metastasis using transcriptional profiling and bioinformatics analysis of ccRCC samples, and to elucidate the role of FOXO3a in ccRCC metastasis.Experimental Design: Gene expression profiling was performed using four primary metastatic and five primary nonmetastatic ccRCC samples. The mRNA and protein levels of FOXO3a in ccRCC samples were investigated by real-time reverse transcription PCR and immunohistochemistry, respectively. The association between metastasis-free survival of patients with ccRCC and FOXO3a mRNA levels was analyzed. Biologic functions of FOXO3a in renal cancer cell lines were investigated. The influence of FOXO3a on tumor metastasis was also studied in vivo orthotopic xenograft tumor model. Finally, the mechanism by which FOXO3a attenuation could increase invasion and migration of tumor cells was explored.Results: Bioinformatics analysis of the profiling data identified FOXO3a as a key factor in ccRCC metastasis. FOXO3a expression was decreased in primary metastatic ccRCC samples. Patients with low FOXO3a mRNA levels had poor metastasis-free survival (P = 0.003). Knocking down FOXO3a induced tumor cell invasion and migration in the nonmetastatic ccRCC cells. Induced FOXO3a overexpression in SN12-PM6 cells could inhibit tumor metastasis in vivo. Downregulation of FOXO3a increased SNAIL1 expression, thereby activating the epithelial–mesenchymal transition (EMT) of RCC cell lines.Conclusions: The loss of FOXO3a induced EMT of tumor cells by upregulating SNAIL1, which promoted tumor cells metastasis in vitro and in vivo. Thus, FOXO3a could be considered as an independent prognostic factor in ccRCC metastasis and could be a marker of occult metastases. Clin Cancer Res; 20(7); 1779–90. ©2014 AACR.

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PDF file - 143KB, Pathway analysis based on KEGG, Biocarta, and Reatome database.

PDF file - 136KB, Analysis of expression profiling data for filtering central genes.

PDF file - 2206KB, Scratch wound migration assays in the 786-O and SN12-PM6 cells.

Objectives: To compare the perioperative hemodynamic consequences and oncology outcomes of robotic retroperitoneal vs transperitoneal inferior vena cava (IVC) thrombectomy (IVCT) for right...

The classical motor center cerebellum is one of the most consistent structures of abnormality in autism spectrum disorders (ASD), and neuropeptide oxytocin is increasingly explored as a potential pharmacotherapy for ASD. However, whether oxytocin targets the cerebellum for therapeutic effects remains unclear. Here, we report a localization of oxytocin receptor (OXTR) in Purkinje cells (PCs) of cerebellar lobule Crus I, which is functionally connected with ASD-implicated circuits. OXTR activation neither affects firing activities, intrinsic excitability, and synaptic transmission of normal PCs nor improves abnormal intrinsic excitability and synaptic transmission of PCs in maternal immune activation (MIA) mouse model of autism. Furthermore, blockage of OXTR in Crus I in wild-type mice does not induce autistic-like social, stereotypic, cognitive, and anxiety-like behaviors. These results suggest that oxytocin signaling in Crus I PCs seems to be uninvolved in ASD pathophysiology, and contribute to understanding of targets and mechanisms of oxytocin in ASD treatment.

Betahistine and gastrodin are the first-line medications for vestibular disorders in clinical practice, nevertheless, their amelioration effects on vestibular dysfunctions still lack direct comparison and their unexpected extra-vestibular effects remain elusive. Recent clinical studies have indicated that both of them may have effects on the gastrointestinal (GI) tract. Therefore, we purposed to systematically compare both vestibular and GI effects induced by betahistine and gastrodin and tried to elucidate the mechanisms underlying their GI effects. Our results showed that betahistine and gastrodin indeed had similar therapeutic effects on vestibular-associated motor dysfunction induced by unilateral labyrinthectomy. However, betahistine reduced total GI motility with gastric hypomotility and colonic hypermotility, whereas gastrodin did not influence total GI motility with only slight colonic hypermotility. In addition, betahistine, at normal dosages, induced a slight injury of gastric mucosa. These GI effects may be due to the different effects of betahistine and gastrodin on substance P and vasoactive intestinal peptide secretion in stomach and/or colon, and agonistic/anatgonistic effects of betahistine on histamine H1 and H3 receptors expressed in GI mucosal cells and H3 receptors distributed on nerves within the myenteric and submucosal plexuses. Furthermore, treatment of betahistine and gastrodin had potential effects on gut microbiota composition, which could lead to changes in host-microbiota homeostasis in turn. These results demonstrate that gastrodin has a consistent improvement effect on vestibular functions compared with betahistine but less effect on GI functions and gut microbiota, suggesting that gastrodin may be more suitable for vestibular disease patients with GI dysfunction.

The subthalamic nucleus (STN) is an effective therapeutic target for deep brain stimulation (DBS) for Parkinson's disease (PD), and histamine levels are elevated in the basal ganglia in PD patients. However, the effect of endogenous histaminergic modulation on STN neuronal activities and the neuronal mechanism underlying STN-DBS are unknown. Here, we report that STN neuronal firing patterns are more crucial than firing rates for motor control. Histamine excited STN neurons, but paradoxically ameliorated parkinsonian motor deficits, which we attributed to regularizing firing patterns of STN neurons via the hyperpolarization-activated cyclic nucleotide-gated channel 2 (HCN2) channel coupled to the H2 receptor. Intriguingly, DBS increased histamine release in the STN and regularized STN neuronal firing patterns under parkinsonian conditions. HCN2 contributed to the DBS-induced regularization of neuronal firing patterns, suppression of excessive β oscillations, and alleviation of motor deficits in PD. The results reveal an indispensable role for regularizing STN neuronal firing patterns in amelioration of parkinsonian motor dysfunction and a functional compensation for histamine in parkinsonian basal ganglia circuitry. The findings provide insights into mechanisms of STN-DBS as well as potential therapeutic targets and STN-DBS strategies for PD.

The number of patients with adrenal aldosterone-producing adenomas (APAs) has gradually increased. However, even after adenoma resection, some patients still suffer from high systolic blood pressure (SBP), which is possibly due to great arterial remodeling. Moreover, mineralocorticoid receptors (MRs) were found to be expressed in vascular smooth muscle cells (VSMCs). This study aims to determine whether MR antagonism protects the aorta from aldosterone-induced aortic remolding. Male rats were subcutaneously implanted with an osmotic minipumps and randomly divided into four groups: control; aldosterone (1 μg/h); aldosterone plus a specific MR antagonist, eplerenone (100 mg/kg/day); and aldosterone plus a vasodilator, hydralazine (25 mg/kg/day). After 8 weeks of infusion, aortic smooth muscle cell proliferation and collagen deposition, as well as the MDM2 and TGF-β1 expression levels in the aorta, were examined. Model rats with APAs were successfully constructed. Compared with the control rats, the model rats exhibited (1) marked SBP elevation, (2) no significant alteration in aortic morphology, (3) increased VSMC proliferation and MDM2 expression in the aorta, and (4) enhanced total collagen and collagen III depositions in the aorta, accompanied with up-regulated expression of TGF-β1. These effects were significantly inhibited by co-administration with eplerenone but not with hydralazine. These findings suggested that specific MR antagonism protects the aorta from aldosterone-induced VSMC proliferation and collagen deposition.

The present study aimed to investigate presurgical neoadjuvant targeted therapy for patients with kidney cancer and vena cava tumor embolus, in order to examine its indications, therapeutic effects and optimal timing of surgery. Between June 2009 and June 2014, 12 patients from The People's Liberation Army General Hospital (Beijing, China) were diagnosed with kidney cancer with superior vena cava tumor embolus, and received presurgical neoadjuvant targeted therapy (sorafenib 400 mg twice a day or sunitinib 50 mg/day) for a median of 13.3 weeks. Patients included 8 males and 4 females, with a median age of 49.8 years. Kidney cancer was present on the left side in 3 patients and in the right side in 9 patients. The median tumor embolus length was 9.7 cm (range, 6.5–14.0 cm). Tumor embolus levels II, III and IV, classified by the Mayo Clinic standard, were observed in 2, 6 and 4 patients, respectively. Median treatment time and average targeted therapy discontinuation time were observed to be longer in sunitinib-treated patients compared with sorafenib-treated patients. In total, 2 patients in the present study had partial remission (PR) and 8 patients had stable disease (SD); for tumor embolus, 4 patients had PR and 8 patients had SD. Tumor embolus length decreased by a median value of 18.7% (range, 0.0–42.1%) or 1.8 cm (range, 0.1–5.2 cm). Tumor diameter decreased by a median value of 8.6% (range, 0.0–38.9%) or 0.7 cm (range, 0.0–3.5 cm). The level of the tumor thrombus, classified by the Mayo Clinic standard, was observed to decrease following sunitinib treatment, including two cases downgraded from tumor thrombus level IV to II, one case from level IV to III and two cases from level III to II. Presurgical neoadjuvant targeted molecular therapy may have the potential to reduce the tumor stage of patients, as well as decreasing the surgical difficulty for radical nephrectomy.

Spinal α-motoneurons directly innervate skeletal muscles and function as the final common path for movement and behavior. The processes that determine the excitability of motoneurons are critical for the execution of motor behavior. In fact, it has been noted that spinal motoneurons receive various neuromodulatory inputs, especially monoaminergic one. However, the roles of histamine and hypothalamic histaminergic innervation on spinal motoneurons and the underlying ionic mechanisms are still largely unknown. In the present study, by using the method of intracellular recording on rat spinal slices, we found that activation of either H

Objective: The aim of this study was to explore a new treatment strategy for left renal vein tumor thrombus directed at the thrombus level and the therapeutic effect of robotic surgery. Ma...

To evaluate the application of laparoscopic extended partial cystectomy (PC) and bilateral extended pelvic lymph node dissection (PLND) for the treatment of urachal carcinomas (UrCs).Combined laparoscopic extended PC and bilateral extended PLND was performed in 16 cases with UrCs in our hospital between April 2009 and December 2012. The surgical procedure included the dilation of the extraperitoneal space, circumscription of the umbilicus, dissection from the umbilicus caudad to the dome of the bladder, excision of the tumor, and the suture of the bladder. The average age of the patients was 52.8 years (35-73 years). The average tumor size was 3.2 cm (1.5-5.6 cm). A median follow-up period of 36 months was obtained to evaluate patient survival and recrudescence.All 16 procedures were completed laparoscopically without open conversion. The median operation time was 85 minutes (65-125 minutes), with a median estimated blood loss of 50 mL (30-110 mL). The median hospital stay was 5 days (4-7 days). The bladder margins were negative in all cases. However, five cases were confirmed postoperatively with positive lymph nodes. No intraoperative or postoperative complications occurred. Histopathology confirmed mucous urachal adenocarcinoma in 11 cases, mixed carcinoma in 1 case, and papillary adenocarcinoma in 4 cases. After a median follow-up period of 36 months, the 2- and 3-year survival rates were 62.5% (10/16) and 50% (8/16), respectively.Combined laparoscopic extended PC and bilateral extended PLND is a safe and feasible method for treating patients with urachal malignancy.

Renal cell carcinoma (RCC) is the most common malignancy involving the kidneys and a major cause of cancer mortality. The involvement of microRNA (miRNA) expression in the tumorigenesis and progression of RCC has been previously highlighted. Therefore, we conducted this study to investigate whether microRNA-363 (miR-363) affects the development of RCC via the Janus tyrosine kinases (JAK2)-signal transducers and activators of transcription (STAT) axis by targeting the growth hormone receptor (GHR), by observing the changes that occurred in the RCC and the normal adjacent tissues of patients with RCC. RCC cells were transfected with a series of miR-363 mimic, miR-363 inhibitor, or small interfering RNA against GHR to determine the influence of miR-363 on the expression of GHR and JAK2-STAT3 axis-related genes with the use of reverse transcription quantitative polymerase chain reaction and Western blot analysis. The angiogenesis, viability, invasion, and migration of cells were evaluated by means of in vitro angiogenesis, 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT), wound-healing, and Transwell assays. The results revealed reduced miR-363 expression and elevated GHR expression in RCC. It was also found that miR-363 altered the activation of the JAK2-STAT3 axis through the inhibition of GHR. Cells treated with the miR-363 inhibitor presented with increased capillary vessels, cell viability, invasion, and migration, whereas it was on the contrary in the RCC cells with overexpressed miR-363. These results implicated that the overexpression of miR-363 could specifically bind to GHR to downregulate the expression of GHR, which, in turn, inactivates the JAK2-STAT3 axis, thereby influencing the angiogenesis, cell invasion, and migration abilities in RCC.

BACKGROUND Current studies indicated that PCDH17 functions as a tumor suppressor, which is frequently inactivated by aberrant promoter methylation in urologic tumors. The main purpose of this study was to investigate the methylation status of PCDH17 in serum and its clinical significance in renal cell carcinoma (RCC). MATERIAL AND METHODS The methylation status of PCDH17 in serum samples of 142 RCC patients and 34 controls was evaluated by methylation-specific PCR (MSP). Then we correlated PCDH17 methylation status with the clinicopathologic features of RCC patients and patient outcomes. RESULTS We found that PCDH17 was more frequently methylated in RCC patients than in controls. Moreover, PCDH17 methylation in serum was significantly correlated with advanced stage (p=0.044), higher grade (p=0.019), lymph node metastasis (p=0.008) and tumor progression (p

Cerebellar ataxia, characterized by motor incoordination, postural instability, and gait abnormality [1-3], greatly affects daily activities and quality of life. Although accumulating genetic and non-genetic etiological factors have been revealed [4-7], effective therapies for cerebellar ataxia are still lacking. Intriguingly, corticotropin-releasing factor (CRF), a peptide hormone and neurotransmitter [8, 9], is considered a putative neurotransmitter in the olivo-cerebellar system [10-14]. Notably, decreased levels of CRF in the inferior olive (IO), the sole origin of cerebellar climbing fibers, have been reported in patients with spinocerebellar degeneration or olivopontocerebellar atrophy [15, 16], yet little is known about the exact role of CRF in cerebellar motor coordination and ataxia. Here we report that deficiency of CRF in the olivo-cerebellar system induces ataxia-like motor abnormalities. CRFergic neurons in the IO project directly to the cerebellar nuclei, the ultimate integration and output node of the cerebellum, and CRF selectively excites glutamatergic projection neurons rather than GABAergic neurons in the cerebellar interpositus nucleus (IN) via two CRF receptors, CRFR1 and CRFR2, and their downstream inward rectifier K+ channel and/or hyperpolarization-activated cyclic nucleotide-gated (HCN) channel. Furthermore, CRF promotes cerebellar motor coordination and rescues ataxic motor deficits. The findings define a previously unknown role for CRF in the olivo-cerebellar system in the control of gait, posture, and motor coordination, and provide new insight into the etiology, pathophysiology, and treatment strategy of cerebellar ataxia.

The tailor welded blank stamping is a complicated process with numerous forming parameters, so it is difficult to determine the relationship between the forming parameters and the formability. The TWBs forming is numerically simulated by an orthogonal experiment design while the thickness ratio, blank holder force and frictional coefficient are forming parameters, and the maximum thickness thinning and the maximum weld movement are index. The influencing degree of forming parameters on the index and the optimal combination of forming parameters are analyzed. The optimization design is used to carry out numerical simulation and verification which can effectively improve stamping formability of TWBs. The simulation results also show that optimization schemes can offer the reference for the forming parameters optimization of the TWBs.

Purpose: To explore the mechanisms underlying clear-cell renal cell carcinoma (ccRCC) metastasis using transcriptional profiling and bioinformatics analysis of ccRCC samples, and to elucidate the role of FOXO3a in ccRCC metastasis. Experimental Design: Gene expression profiling was performed using four primary metastatic and five primary nonmetastatic ccRCC samples. The mRNA and protein levels of FOXO3a in ccRCC samples were investigated by real-time reverse transcription PCR and immunohistochemistry, respectively. The association between metastasis-free survival of patients with ccRCC and FOXO3a mRNA levels was analyzed. Biologic functions of FOXO3a in renal cancer cell lines were investigated. The influence of FOXO3a on tumor metastasis was also studied in vivo orthotopic xenograft tumor model. Finally, the mechanism by which FOXO3a attenuation could increase invasion and migration of tumor cells was explored. Results: Bioinformatics analysis of the profiling data identified FOXO3a as a key factor in ccRCC metastasis. FOXO3a expression was decreased in primary metastatic ccRCC samples. Patients with low FOXO3a mRNA levels had poor metastasis-free survival (P = 0.003). Knocking down FOXO3a induced tumor cell invasion and migration in the nonmetastatic ccRCC cells. Induced FOXO3a overexpression in SN12-PM6 cells could inhibit tumor metastasis in vivo. Downregulation of FOXO3a increased SNAIL1 expression, thereby activating the epithelial–mesenchymal transition (EMT) of RCC cell lines. Conclusions: The loss of FOXO3a induced EMT of tumor cells by upregulating SNAIL1, which promoted tumor cells metastasis in vitro and in vivo. Thus, FOXO3a could be considered as an independent prognostic factor in ccRCC metastasis and could be a marker of occult metastases. Clin Cancer Res; 20(7); 1779–90. ©2014 AACR.

// Qing Bo Huang 1,* , Xin Ma 1,* , Hong Zhao Li 1 , Qing Ai 1 , Shang Wen Liu 2 , Yu Zhang 1 , Yu Gao 1 , Yang Fan 1 , Dong Ni 3 , Bao Jun Wang 1 and Xu Zhang 1 1 Department of Urology/State Key Laboratory of Kidney Diseases, Chinese PLA General Hospital/PLA Medical School, Beijing, China 2 Department of Urology, Chinese PLA 303 Hospital, Nanning, China. 3 Department of Urology, Zhongnan Hospital of Wuhan University, Wuhan, China. Correspondence: Xu Zhang, email: // Keywords : Kidney neoplasms, Neoplasm metastasis, Cell communication, Signal transduction, Angiogenesis Received : January 2, 2014 Accepted : March 14, 2014 Published : March 14, 2014 Abstract The Notch ligand Delta-like 4 (DLL4) plays an important role in tumor angiogenesis, which is required for tumor invasion and metastasis. Here we showed that DLL4 was elevated in endothelium and Notch signaling was activated in renal cell carcinoma (RCC). Exogenous DLL4 induced RCC cell migration and invasion by activating intercellular Notch signaling. Importantly, the DLL4/Notch/Hey1/MMP9 cascades connecting the endothelium to the cancer cells in metastasis were identified. Knockdown of Hey1 decreased expression of MMP9 and attenuated tumor invasion. The clinical investigation on 120 cases of RCC specimens indicated that expressions of Hey1 and MMP9 correlated with DLL4 density. Moreover, univariate and multivariate analyses showed that tumor hematogenous metastasis not only was depended on microvessel density but was also associated with tumor size and DLL4 density. During 4-year surveillance, high-level of DLL4 density was associated with a higher probability of developing metastasis and being sensitive to target therapies. Our data suggest that RCC progression is caused in part by activated DLL4/Notch signaling, interaction of endothelium and cells, which can be therapeutically targeted.

The aim of this study was to validate the advantages of the intrafascial nerve-sparing technique compared with the interfascial nerve-sparing technique in extraperitoneal laparoscopic radical prostatectomy. From March 2010 to August 2011, 65 patients with localized prostate cancer (PCa) underwent bilateral intrafascial nerve-sparing extraperitoneal laparoscopic radical prostatectomy. These patients were matched in a 1:2 ratio to 130 patients with localized PCa who had undergone bilateral interfascial nerve-sparing extraperitoneal laparoscopic radical prostatectomy between January 2008 and August 2011. Operative data and oncological and functional results of both groups were compared. There was no difference in operative data, pathological stages and overall rates of positive surgical margins between the groups. There were 9 and 13 patients lost to follow-up in the intrafascial group and interfascial group, respectively. The intrafascial technique provided earlier recovery of continence at both 3 and 6 months than the interfascial technique. Equal results in terms of continence were found in both groups at 12 months. Better rates of potency at 6 months and 12 months were found in younger patients (age ≤ 65 years) and overall patients who had undergone the intrafascial nerve-sparing extraperitoneal laparoscopic radical prostatectomy. Biochemical progression-free survival rates 1 year postoperatively were similar in both groups. Using strict indications, compared with the interfascial nerve-sparing technique, the intrafascial technique provided similar operative outcomes and short-term oncological results, quicker recovery of continence and better potency. The intrafascial nerve-sparing technique is recommended as a preferred approach for young PCa patients who are clinical stages cT1 to cT2a and have normal preoperative potency.

Purpose: The purpose of this study was to introduce a new method for locating the renal artery during retroperitoneal laparoscopic renal surgery. Patients and Methods: The medial arcuate ligament (MAL) is a tendinous arch in the fascia under the diaphragm that arches across the psoas major muscle and is attached medially to the side of the first or the second lumbar vertebra. The renal artery arises at the level of the intervertebral disc between the L1 and L2 vertebrae. We evaluate the role of the MAL that serves as an anatomic landmark for locating the renal artery during retroperitoneal laparoscopic renal surgery. Results: There is a reproducible consistent anatomic relationship between MAL and the renal artery in 210 cases of retroperitoneal laparoscopic renal surgery. Two main types of the MAL, the “narrow arch” and the “fascial band” types, can be observed. Conclusion: MAL can serve as an accurate and reproducible anatomic landmark for the identification of the renal artery during retroperi...

Corticotropin releasing factor (CRF), a peptide hormone involved in the stress response, holds a key position in cardiovascular regulation. Here, we report that the central effect of CRF on cardiovascular activities is mediated by the posterior hypothalamic nucleus (PH), an important structure responsible for stress-induced cardiovascular changes. Our present results demonstrate that CRF directly excites PH neurons via two CRF receptors, CRFR1 and CRFR2 and consequently increases heart rate (HR) rather than the mean arterial pressure (MAP) and renal sympathetic nerve activity (RSNA). Bilateral vagotomy does not influence the tachycardia response to microinjection of CRF into the PH, while β adrenergic receptor antagonist propranolol almost totally abolishes the tachycardia. Furthermore, microinjecting CRF into the PH primarily increases neuronal activity of the rostral ventrolateral medulla (RVLM) and rostral ventromedial medulla (RVMM), but does not influence that of the dorsal motor nucleus of the vagus nerve (DMNV). These findings suggest that the PH is a critical target for central CRF system in regulation of cardiac activity and the PH-RVLM/RVMM-cardiac sympathetic nerve pathways, rather than PH-DMNV-vagus pathway, may contribute to the CRF-induced tachycardia.

Objective To explore the correlation between microRNA-34a (miR-34a) and Notch1, and evaluate the influence of miR-34a overexpression on proliferation of bladder cancer cell line T24. Methods Bioinformatics software were used for predicting the binding site of miR-34a and Notch1, and luciferase assay was performed for confirming the direct regulatory relationship between them. miR-34a plasmid was transfected into bladder cancer cell line T24. Notch1 expression was detected by quantitative real-time polymerase chain reaction and Western blotting. Cell proliferation was assayed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Apoptosis and cell cycle were assessed by flow cytometry. Results Luciferase assays showed that miR-34a transfection significantly down-regulated the normalized Notch1 3'UTR luciferase activity (P=0.006). Transfection of miR-34a reduced the RNA and protein levels of Notch1. Cell proliferation assay revealed that miR-34a transfection suppressed the proliferation of T24 cells in a time-dependent manner (P < 0.001). Ectopic expression of miR-34a caused significant increase of apoptotic cells (P=0.003) and induced cell cycle arrest at G0-G1 phase in T24 cells (P=0.002). Conclusion Overexpressed miRNA-34a can inhibit proliferation of bladder cancer cells by antagonizing Notch1, thus indicating its tumor-suppressive function in bladder cancer.

To our knowledge we present the initial experience with and the short-term outcome of laparo-endoscopic single site anatomical retroperitoneoscopic adrenalectomy using conventional instruments.Between June 2009 and April 2010, 25 patients underwent laparo-endoscopic single site anatomical retroperitoneoscopic adrenalectomy. A TriPort™ Access System was inserted through a 2.5 to 3.0 cm transverse skin incision below the tip of the 12th rib. Adrenalectomy was done using a 5 mm 30-degree laparoscopic camera and 2 conventional laparoscopic instruments. After Gerota's fascia was incised we explored the first dissection plane between the perirenal fat and the anterior renal fascia at the superomedial side of the kidney. The adrenal gland was identified at the initial stage of the operation.Laparo-endoscopic single site anatomical retroperitoneoscopic adrenalectomy was successfully accomplished in 23 patients. An additional 5 mm port was required in 1 of the 2 unsuccessful cases and in the other it was necessary to convert to standard anatomical retroperitoneoscopic adrenalectomy. Median incision length was 3.0 cm, median operative time was 55 minutes and median estimated blood loss was 15 ml. No major intraoperative complications occurred. In the initial 10 cases median operative time was significantly longer (62 vs 50 minutes) and median blood loss was significantly higher (75 vs 10, each p0.001) than in the subsequent 15. In 3 cases pheochromocytoma was successfully excised without undesirable hemodynamic oscillation. Postoperative complications developed in 2 patients, including angina and contralateral atelectasis in 1 each.In properly selected patients laparo-endoscopic single site anatomical retroperitoneoscopic adrenalectomy with conventional instruments is feasible, safe and effective, causes minimal morbidity and results in excellent cosmesis.

To develop a new porcine model with horn of the uterus to mimic an enlarged ureter for training for laparoscopic ureteral reimplantation (LUR) and to evaluate its feasibility.Ten female pigs were used in the training. The pig was placed to a dorsal position after an anesthetic was administered. The horn of the uterus near the bladder was dissected, then spatulated and trimmed to replace the enlarged ureter. LUR was performed according to standard operation steps. Four trainees completed the LUR procedure based on a mentor-trainee model to guarantee the success of the procedure and the quality of the anastomoses. The learning curve of operative time was analyzed. The anastomotic stoma was cut off postoperatively and checked extracorporeally. After the course, questionnaire surveys were sent to the trainees to investigate satisfaction of the training and assess the impact of the training on their learning of "real" LUR in future practice.This model reproduced the key technique steps of LUR. Four LUR procedures were performed on each pig. The operative time declined from 170.0 +/- 10.3 minutes to 90.3 +/- 3.7 minutes (P0.01) after the trainees had performed 10 LURs. There was proper stitching in each "ureterovesical" anastomosis. At the end of training, all trainees could accomplish a LUR procedure skillfully on the model; they were satisfied after the course and thought the training was helpful to future practice of LUR.The new model was feasible and cost-effective for training in the basic skills of laparoscopic ureteral reconstruction procedures.

Raf kinase inhibitor protein (RKIP) has been shown to be a metastasis suppressor in many kinds of malignant tumors. But its function in breast cancer was not yet clarified completely. We detected RKIP expression in clinical samples of primary breast cancer, breast cancer metastases, and in different breast cancer cells. Compared with the normal breast epithelia, benign breast epithelia, or in situ ductal carcinoma, the expression level of RKIP is decreased in invasive carcinoma and significantly reduced or lost in the metastasis lymph node matched to the invasive carcinoma. To explore the potential role of RKIP in breast cancer metastasis, we studied the effect of RKIP on the malignant phenotypes of the breast cancer cells with ectopically overexpression or knockdown of RKIP. Cell proliferation, soft-agar colony formation, in vitro adhesion assay, invasion, and migation assays were done to examine the malignant phenotypes of the transfected cells. Consequently, RKIP has no effect on in vitro proliferation rate or colony-forming ability of MDA-MB-435 cells. In vitro cell invasion and migration assays indicated that the RKIP expression was inversely associated with the invasiveness of MDA-MB-435 cells. Consistent with these results, in the orthotopic murine models, we observed that overexpression of RKIP in breast cancer cells impaired invasiveness and metastasis, whereas down-regulation of RKIP expression promoted invasiveness and metastasis. These results indicate that RKIP is a metastasis suppressor gene of human breast cancer. (Mol Cancer Res 2009;7(6):832–40)

Through whole-cell patch recordings in brainstem slices, the effects of histamine on neuronal activity of the lateral vestibular nucleus (LVN) were investigated. Bath application of histamine elicited a concentration-dependent excitation of both spontaneous firing (n=19) and silent (n=7) LVN neurons. Moreover, histamine induced a stable inward current in the LVN neurons (n=5) and the histamine-induced depolarization of membrane potential persisted in the presence of tetrodotoxin (n=4), indicating a direct post-synaptic effect of the histamine on the LVN neurons. Selective histamine H2 receptor antagonist ranitidine effectively blocked the histamine-evoked excitatory responses on the LVN neurons (n=4), but selective histamine H1 receptor antagonist triprolidine did not (n=4). In addition, selective histamine H2 receptor agonist dimaprit (n=3) rather than 2-pyridylethylamine (n=4), a selective histamine H1 receptor agonist, mimicked the excitatory action of histamine on LVN neurons. The results demonstrate that histamine excites the LVN neurons via post-synaptic histamine H2 receptors and suggest that the central histaminergic projection arising from the hypothalamus may modulate LVN neurons activity and actively influence the vestibular reflexes and functions.

Loss of function of metastasis suppressor genes is an important step in the progression to a malignant tumor type. Studies in cell culture and animal models have suggested a role of Raf kinase inhibitor protein (RKIP) in suppressing the metastatic spread of prostate cancer, breast cancer, and melanoma cells. However, the function of RKIP in ovarian cancer (OVCA) has not been reported. To explore the potential role of RKIP in epithelial OVCA metastasis, we detected the expression levels of RKIP protein in tissue samples from patients with epithelial OVCA. Consequently, the expression of RKIP is reduced in the poorly differentiated OVCA than in the well-differentiated and moderately differentiated OVCA. In addition, in vitro cell invasion assay indicated that the RKIP expression was inversely associated with the invasiveness of five OVCA cell lines. Consistent with this result, the cell proliferation, anchorage-independent growth, cell adhesion, and invasion were decreased in RKIP overexpressed cells but increased in RKIP down-regulated cells. Further investigation indicated that RKIP inhibited OVCA cell proliferation by altering cell cycle progression rather than promoting apoptosis. Furthermore, the overexpression of RKIP suppressed the ability of human OVCA cells to metastasize when the tumor cells were transplanted into nude mice. Our data show the effect of RKIP on the proliferation, migration, or adhesion of OVCA cells. These results indicate that RKIP is also a metastasis suppressor gene of human epithelial OVCA. (Mol Cancer Res 2008;6(6):917–28)

The cerebellar fastigial nucleus (FN) holds an important role in motor control and body balance. Previous studies have revealed that the nucleus is innervated by direct hypothalamocerebellar histaminergic fibers. However, the functional role of histaminergic projection in cerebellar FN has never been established. In this study, we investigated the effect of histamine on neuronal firing of cerebellar FN by using slice preparations. Sixty-five FN cells were recorded from 47 cerebellar slices, and a vast majority of the cells responded to histamine stimulation with an excitatory response (58/65, 89.2%). Perfusing slices with low-Ca2+/high-Mg2+ medium did not block the histamine-induced excitation (n=10), supporting a direct postsynaptic action of histamine on the cells. Furthermore, the excitatory effect of histamine on FN neurons was not blocked by selective histamine H1 receptor antagonist triprolidine (n=15) or chlorpheniramine (n=10), but was effectively suppressed by ranitidine (n=15), a highly selective histamine H2 receptor antagonist. On the other hand, highly selective histamine H2 receptor agonist dimaprit (n=20) instead of histamine H1 receptor agonist 2-pyridylethylamine (n=16) mimicked the excitatory effect of histamine on FN neurons. The dimaprit-induced FN neuronal excitation was effectively antagonized by selective histamine H2 receptor antagonist ranitidine (n=13) but not influenced by selective histamine H1 receptor antagonist triprolidine (n=15). These results demonstrate that histamine excites cerebellar FN cells via the histamine H2 receptor mechanism and suggest that the hypothalamocerebellar histaminergic fibers may modulate cerebellar FN-mediated sensorimotor integration through their excitatory innervations on FN neurons.

Previous studies have revealed a direct histaminergic projection from the tuberomamillary nucleus of hypothalamus to the cerebellum and a postsynaptic excitatory effect of histamine on the cerebellar interpositus nucleus neurons via histamine H 2 receptors in vitro , indicating that the histaminergic afferent inputs of cerebellar nuclei may be involved in the cerebellar function of motor control. To test this hypothesis, in this study histaminergic agents were bilaterally microinjected into the cerebellar interpositus nucleus of intact adult male rats, and their effects on motor balance and coordination of the animals performing accelerating rota-rod treadmill and balance beam tasks were observed. The results showed that microinjection of histamine into the cerebellar interpositus nucleus remarkably increased the time that animals balanced steadily on the rota-rod and markedly shortened the duration of passage through the balance beam, whereas GABA significantly depressed motor performances of animals on the rota-rod and beam, and normal saline influenced neither. In addition, administration of selective histamine H 2 receptor antagonist ranitidine considerably decreased the animals’ endurance time on rota-rod and noticeably increased the passing time on beam, but selective histamine H 1 receptor antagonist triprolidine showed no effect. Furthermore, microinjection of histamine reversed the inhibitory effects of ranitidine on rota-rod and beam performance. These results demonstrate that histamine enhances rat motor balance and coordination through activation of histamine H 2 receptors in the cerebellar interpositus nucleus and suggest that the hypothalamocerebellar histaminergic projections may play a modulatory role on the cerebellar circuitry to ensure that movements are accurately executed.

We present our experience with retroperitoneoscopic renal pedicle lymphatic disconnection. We compared the clinical efficacy of this treatment for chyluria with that of open surgery.From January 1998 to June 2004, 53 patients (55 renal units) with chyluria underwent renal pedicle lymphatic disconnection via the retroperitoneoscopic and conventional open approaches. The diagnosis of chyluria was confirmed by the ether test and the side of chylous reflux was determined by cystoscopy. Operative time, intraoperative blood loss, postoperative intestinal recovery and hospital stay were evaluated. Increases in hemoglobin and serum albumin were compared before and after surgery during followup.Retroperitoneoscopic renal pedicle lymphatic disconnection or open surgery was performed successfully in all patients. In terms of operative time, intraoperative blood loss, postoperative intestinal recovery and hospital stay retroperitoneoscopy was superior to conventional open surgery. During retroperitoneoscopy the inferior vena cava was injured in 1 case but repaired successfully by laparoscopy without conversion to open surgery. Postoperative gross hematuria in 1 case disappeared 4 days later. In the open surgery group the renal segmental artery was inadvertently injured in 1 case and anastomosis was performed successfully. Wound healing was delayed in 1 case due to hypoalbuminemia. Recurrence developed in 2 patients during the 6 to 84-month followup.Retroperitoneoscopic renal pedicle lymphatic disconnection for chyluria has the advantages of minimal invasion and rapid recovery compared with open surgery.

Previous studies suggested that the postsynaptic GABA B receptors of deep cerebellar nuclear neurons of adult rats were not activated by selective GABA B receptor agonist baclofen or endogenous GABA released by cerebellar cortical Purkinje cells, although the receptors have been demonstrated to exist in the deep cerebellar nuclei. In this study, cerebellar slices of adult rats were prepared for testing effects of GABA, baclofen and muscimol (selective GABA A receptor agonist) on cerebellar interpositus nucleus (IN) neurons. Perfusing slices with GABA (10–1000 μM), baclofen (1–30 μM) and muscimol (1–100 μM) respectively produced a dose-dependent inhibitory response on the IN neurons ( n = 39, 62 and 50), which was not blocked by low-Ca 2+ /high-Mg 2+ medium ( n = 5, 6 and 6), supporting a direct postsynaptic action of these GABAergic agonists. Moreover, both selective GABA B receptor antagonist CGP35348 and selective GABA A receptor antagonist bicuculline were capable of partially blocking the inhibitory response of IN neurons to GABA ( n = 14 and 11), suggesting that the GABA-induced inhibition may contain two components, a GABA B receptors-mediated component and a GABA A receptors-mediated one. Further experiments revealed that not only muscimol ( n = 50) but also baclofen ( n = 62) suppressed IN cells’ activity. The baclofen-induced inhibition was selectively blocked by CGP35348 ( n = 12) but not by bicuculline ( n = 8), whereas the muscimol-induced inhibition was selectively antagonized by bicuculline ( n = 8) instead of CGP35348 ( n = 9). These results indicate that GABA B receptors in the IN neurons can be activated not only by GABA but also by baclofen, suggesting that besides GABA A receptors, GABA B receptors may also be involved in mediating the inhibitory effect of GABA on cerebellar IN neurons of adult rats.

Previous investigations have demonstrated that the neuronal activity in the lateral hypothalamic area (LHA) is respectively modulated by afferent inputs from the gastric vagal nerves innervating the upper gastrointestinal tract, as well as the cerebellar interpositus nucleus (IN). The aim of this study was to examine whether the gastric vagal and cerebellar IN inputs converge onto single LHA neurons in rats, especially those sensitive to glycemia. Of the 114 LHA neurons recorded, 60 (52.6%) and 51 (44.7%) responded to gastric vagal and cerebellar IN stimulation, respectively. Of the 60 LHA neurons responsive to gastric vagal stimulation, 30 also responded to the cerebellar IN stimulus, indicating a convergence of gastric vagal and cerebellar inputs onto single hypothalamic cells. When the gastric vagal nerves and cerebellar IN were stimulated simultaneously, a summation of the responses was observed in all 6 neurons tested. Moreover, of 24 neurons that responded to both the gastric vagal and cerebellar IN stimuli, 15 (62.5%) were identified as glycemia-sensitive. These results demonstrate that the visceral information transmitted by the gastric vagal nerves and the somatic information forwarded by the cerebellar IN converge onto single LHA neurons, especially those sensitive to glycemia. The findings also suggest that integration of somatic-visceral responses related to short-term feeding regulation may take place in the LHA.

Perfusing slices with histamine (1-100 microM) produced an excitatory response in rat rubral neurons (118/132, 89.4%). The histamine-induced excitation was not blocked by the low-Ca2+/high-Mg2+ medium (n=10), supporting a direct postsynaptic action of the amine. Histamine H2 receptor antagonist ranitidine effectively blocked the excitatory response of rubral neurons to histamine (n=26), but H1 receptor antagonist triprolidine did not (n=24). The excitatory effect of histamine could be mimicked by dimaprit, a highly selective H2 receptor agonist (n=24), and the dimaprit-elicited excitation of the rubral neurons could be blocked by ranitidine (n=16), but not by triprolidine (n=9). In addition, H1 receptor agonist 2-pyridylethylamine could not elicit any response in rubral neurons (n=12). These results indicate that histamine excites red nucleus neurons through H2 receptors and suggest that the histaminergic afferent fibers may play an important functional role in the sensorimotor integration through the red nucleus.

Neuroanatomical studies have revealed a direct hypothalamocerebellar histaminergic pathway, and our previous studies have demonstrated an excitatory effect of histamine on granule and Purkinje cells of the cerebellar cortex. In this study, we further investigated the effect of histamine on the neuronal firing of cerebellar interpositus nucleus (IN) by using cerebellar slice preparations. Eighty-seven IN cells were recorded from 38 slices. The vast majority of the cells responded to histamine stimulation with an excitatory response (79/87, 90.8%), and the rest of them showed no reaction (8/87, 9.2%). The histamine-induced excitation was not blocked by application of low-Ca 2+ /high-Mg 2+ medium ( n =8), supporting a direct postsynaptic action of histamine. The histamine H 2 receptor antagonist ranitidine effectively blocked the excitatory response of IN cells to histamine ( n =23), but the histamine H 1 receptor antagonist triprolidine could not significantly block the histamine-induced excitation, or only very slightly decreased the excitatory effect of histamine on the cells ( n =21). On the other hand, the highly selective histamine H 2 receptor agonist dimaprit mimicked the excitatory effect of histamine on IN cells and the dimaprit-induced excitation was also blocked by ranitidine ( n =14). Successively perfusing slices with the medium containing ranitidine and triprolidine, respectively, we found that ranitidine exhibited the same blocking effect on the dimaprit-induced excitation, but triprolidine had no such effect ( n =8). Moreover, the histamine H 1 receptor agonist 2-pyridylethylamine did not show any effect on the IN cells ( n =9). These results demonstrate that histamine excites cerebellar IN cells via the histamine H 2 receptor mechanism. Together with our previous results, we suggest that the hypothalamocerebellar histaminergic fibers may modulate neuronal activities of the cerebellar cortex and deep nuclei in parallel. The significance of the excitatory effect of histamine on the cerebellar nuclear cells is discussed.

Objective To evaluate the clinical effect of laparoscopic ureterolysis in the treatment of retroperitoneal fibrosis,and compare the effectiveness with the therapy of open ureterolysis.Methods The clinical data of 10 patients undergone laparoscopic ureterolysis(LS group) and 13 patients undergone open ureterolysis(OS group) were retrospectively reviewed.A comparison was performed between the two groups in terms of operative preparing time,operating time,estimated blood loss,recovery of intestinal function,ambulation activities after operation,postoperative length of hospital stay,total length of hospital stay and complication.Results The estimated blood loss,recovery of intestinal function,postoperative length of hospital stay,total length of hospital stay in LS group(59.50ml,2.70±0.82d,8.70±1.42d,17.40±5.64d) were all better than those in OS group(100.00ml,3.85±1.07d,11.62±3.18d,23.38±5.45d,P < 0.05).The operative preparing time in LS group was longer than that in OS group(29.25±11.43min vs 19.85±7.36min,P < 0.05).No difference existed between the two groups in the operating time and ambulation activities after operation(P > 0.05).No transfusion was required and no complication occurred in the both groups.With a follow-up of 18 months,no recurrence in situ and no obstruction were observed in the both groups.Conclusions Laparoscopic ureterolysis has the advantages of less blood loss,rapid recovery of intestinal function,shorter hospital stay as compared to that of open ureterolysis.It is therefore a safe and effective surgical procedure in treatment of retroperitoneal fibrosis.

To evaluate renorrhaphy techniques and to analyze surgical outcomes in retroperitoneal laparoscopic partial nephrectomy.A retrospective study from January 2008 to December 2011 analyzed 526 patients with renal tumors in whom renorrhaphy was changed from one layer, interrupted, figure-of-eight (n = 228) suture to two layers, continuous, unknotted (n = 298) suture. All procedures were carried out by the same laparoscopic surgeon (XZ). Patient demographics, tumor characteristics, operative outcomes and perioperative renal function were compared.Median follow up for one layer, interrupted, figure-of-eight suture and two layers, continuous, unknotted suture was 31 and 28 months, respectively. The two layers, continuous, unknotted suture group had shorter warm ischemia time (P = 0.021), faster removal of Jackson-Pratt drains (P = 0.029) and shorter hospital stay (P = 0.037) than the one layer, interrupted, figure-of-eight suture group. There was a trend towards a better preservation of glomerular filtration rates in the two layers, continuous, unknotted suture group (P = 0.045). In a multivariable model, the two layers, continuous, unknotted suture technique was a statistically significant independent predictor of warm ischemia time (P = 0.01), hospital stay (P = 0.001) and estimated glomerular filtration rates (P = 0.043).Two layers, continuous, unknotted suture renorrhaphy allows better outcomes than one layer, interrupted, figure-of-eight suture renorrhaphy in retroperitoneal laparoscopic partial nephrectomy. A longer clinical follow-up evaluation is warranted.

Recent neuroanatomical studies have revealed a direct hypothalamocerebellar histaminergic pathway. However, the functional significance of the histaminergic fibers in the cerebellum is not yet clear. In this study, the effects of histamine on the firing of cerebellar Purkinje cells (PCs) were investigated in vitro. Histamine predominantly produced excitatory (106/111, 95.5%) and in a few cases inhibitory (5/111, 4.5%) responses in PCs. The histamine-induced excitation was not blocked by perfusing the slice with low Ca2+ high/Mg2+ medium (n = 8), supporting a direct postsynaptic action of histamine. The histamine H2 receptor antagonist ranitidine effectively blocked the excitatory response of PCs to histamine (n = 20), but triprolidine, an H1 receptor antagonist, could not significantly block the histamine-induced excitation, or only very slightly decreased the excitatory effect of histamine on the cells (n = 13). On the other hand, the highly selective H2 receptor agonist dimaprit mimicked the excitatory effect of histamine on PCs and this dimaprit-induced excitation was also blocked by ranitidine (n = 20), but not triprolidine (n = 8). However, the H1 receptor agonists betahistine and 2-thiazolylethylamine did not show any effect on the PCs (n = 9 and 14). These results reveal that histamine excites cerebellar PCs via H2 receptors and suggest that the hypothalamocerebellar histaminergic fibers may play an important role in functional activities of the cerebellum.