Hydropenia may accelerates the progression of orthotopic bladder cancer induced by N-methyl-N-nitrosourea by increasing the expression levels of AQP1, AQP3, and AQP4.

BACKGROUND: Increasing evidence has demonstrated aquaporins (AQPs) to be critical players in carcinogenesis. Here, we aimed to explore the role of hydropenia in the progression of bladder cancer (BCa), as well as to assess the expression of AQP1, AQP3, and AQP4 in bladder tissues from hydropenic and N-methyl-N-nitrosourea (MNU)-treated rats. METHODS: An orthotopic BCa model was induced by administering Sprague Dawley rats with MNU. A hydropenic rat model was established by administrating rats with 2/3 of the amount of water given to the control group. At week 8, the rats were sacrificed and their bladder tissues were collected. Then, pathological alterations in the rat bladders were assessed by hematoxylin and eosin staining. The RNA and protein expression levels of AQP1, AQP3, and AQP4 were determined by using qRT-PCR and western blot assays. RESULTS: All of the rats (100%) administrated with MNU developed tumors, of which 5 were large (diameter, 0.5-1.0 cm), 10 were medium (diameter, 0.2-0.5 cm), and 5 were small (diameter, <0.2 cm) in size. The tumors were nodular and cauliflower shaped, with multiple satellite focus, and were accompanied by bleeding, ulcers, stones, and residual urine. Hematoxylin and eosin staining revealed that the bladder mucosa was incomplete, with a large amount of necrotic tissue and obvious leukocytic infiltration. The tumor volume in the MNU + hydropenia group was significantly larger than that in the MNU group. Noticeably, hydropenia exacerbated pathological changes induced by MNU administration. QRT-PCR and western blot analysis revealed that the MNU group, hydropenia group, and MNU + hydropenia group had significantly increased levels of AQP1, AQP3, and AQP4 compared to the control group, with the most dramatic increase seen in the MNU + hydropenia group. CONCLUSIONS: Hydropenia exacerbates pathological alterations induced by MNU in rats with orthotopic BCa by increasing the expression levels of AQP1, AQP3, and AQP4. This study reveals a possible mechanism of the occurrence of BCa.

Needle Adjustment Free (NAF) running suture technique (PAN suture) in laparoscopic partial nephrectomy.

BACKGROUND: It is proposed a new running suture technique called Needle Adjustment Free (NAF) technique, or PAN suture. The efficiency and the safety were evaluated in laparoscopic partial nephrectomy. METHODS: This new running suture technique avoids the Needle Adjustment method used in traditional techniques. The new continuous suture technique (11 patients) was compared with the traditional continuous suture method (33 patients) used in both transperitoneal and retroperitoneal laparoscopic partial nephrectomy (LPN) in terms of suture time (ST), warm ischemia time (WIT), blood loss (BL), open conversion rate and post-op discharge time, post-op bleeding, post-op DVT, ΔGFR (affected side, 3 months post-op). Differences were considered significant when P < 0.05. RESULTS: ST in the PAN suture group was 30.37 ± 16.39 min, which was significant shorter (P = 0.0011) than in the traditional technique group which was 13.68 ± 3.33 min. WIT in the traditional technique group was 28.73 ± 7.89 min, while in the PAN suture group was 20.64 ± 5.04 min, P = 0.0028. The BL in entirety in the traditional technique group was 141.56 ± 155.23 mL, and in the PAN suture group was 43.18 ± 31.17 mL (P = 0.0017). BL in patients without massive bleeding in the traditional technique group was significantly greater than in the PAN suture group at 101.03 ± 68.73 mL versus 43.18 ± 31.17 mL (P = 0.0008). The open conversion rate was 0 % in both groups. There was no significant difference between the two groups in postoperative discharge time, post-op bleeding, post-op DVT, ΔGFR (affected side, 3 months post-op). CONCLUSIONS: The NAF running suture technique, or PAN suture, leading to less ST, WIT and BL, which was shown to be more effective and safer than the traditional technique used for LPN. A further expanded research with larger sample size is needed.

An experimental model of the epithelial to mesenchymal transition and pro-fibrogenesis in urothelial cells related to bladder pain syndrome/interstitial cystitis.

BACKGROUND: Suitable in vitro models are needed to investigate urothelial epithelial to mesenchymal transition (EMT) and pro-fibrogenesis phenotype in bladder pain syndrome/interstitial cystitis (BPS/IC). This study is to establish a novel experimental BPS/IC cell model and explore how different concentrations of tumor necrosis factor (TNF)-α influence the EMT and pro-fibrogenesis phenotype of urothelial cells. METHODS: SV-HUC-1 urothelial cells were cultured with 2, 10, or 50 ng/mL TNF-α to mimic chronic inflammatory stimulation. The EMT and pro-fibrogenesis phenotype, including production of collagen I and pro-fibrosis cytokines, were estimated after 72 h of culture. RESULTS: The bladder urothelial cells of BPS/IC exhibited upregulated vimentin, TNF-α and TNF receptor, downregulated E-cadherin, and increased collagen I. Higher concentrations of TNF-α (10 and 50 ng/mL) produced an obvious mesenchymal morphology, enhanced invasion and migratory capacity, increased expression of vimentin, and decreased expression of E-cadherin. Collagen I was increased in cells treated with 2 and 10 ng/mL TNF-α after 72 h. Secretion of interleukin (IL)-6 and IL-8 was promoted with 10 and 50 ng/mL TNF-α, while that of IL-1ß or transforming growth factor-ß was unaffected. Slug and Smad2 were upregulated by TNF-α after 72 h. The Smad pathway was activated most strongly with 10 ng/mL TNF-α and Slug pathway activation was positively correlated with the concentration of TNF-α. CONCLUSIONS: Sustained 10 ng/mL TNF-α stimulation induced the EMT and pro-fibrogenesis phenotype resembling BPS/IC in SV-HUC-1 cells. Minor inflammatory stimulation induced the pro-fibrogenesis phenotype while severe inflammatory stimulation was more likely to produce significant EMT changes. Different degrees of activation of the Slug and Smad pathways may underlie this phenomenon.

Beforehand transection and suturing (BTS) of the dorsal vascular complex: a novel technique in laparoscopic radical prostatectomy.

BACKGROUND: Beforehand transection and suturing (BTS) of the dorsal vascular complex (DVC), a novel technique in non-neurovascular bundle sparing (NVB-sparing) extraperitoneal laparoscopic radical prostatectomy (eLRP), had been proposed; this study aimed to evaluate this technique in clinical laparoscopic procedures. METHODS: Using this new technique, the DVC was transected and sutured after dissection of the pelvic fascia and before dissection of the prostate, especially before ligation of the bilateral prostatic pedicles. This study retrospectively analyzed the data of 90 non NVB-sparing eLRP patients [traditional technique (n=60) and BTS technique (n=30)]. RESULTS: The surgical time in the BTS technique group was 121.73±24.53 min, which was significantly shorter (P=0.0015) than the traditional technique group (144.12±39.68 min). The calculated blood loss in the traditional technique group was 388.45±232.78 mL, and 264.16±130.70 mL in the BTS technique group (P=0.0016). The estimated blood loss in the traditional technique group was 350.34±311.80 mL, which was significantly greater than the BTS technique group (250.33±145.31 mL, P=0.0422). The transfusion rate in the traditional technique group was significantly greater than the BTS technique group (15.00% vs. 0.00%; P=0.0266). The biochemical recurrence rate in traditional technique group was 48.33%, which was higher than in the BTS group (30.00%) (P=0.0465). There was no significant difference between the 2 groups with respect to the pre-operative hemoglobin (Hb) concentration, pre-operative hematocrit (HCT), post-operative Hb concentration, post-operative HCT, ΔHCT, pre-operative blood volume, rectal perforation, open conversion, apical capsule residue, false suture, post-operative bleeding, urinary leakage, re-operation, surgical site infection, post-operative stay, and emission time of urinary incontinence. CONCLUSIONS: In managing the relationship between the DVC and prostate in patients undergoing non NVB-sparing eLRP, the BTS technique was shown to be more effective and safer than the traditional technique.

Is the 4 mm height of the vertebral artery groove really a limitation of C1 pedicle screw insertion?

PURPOSE: To explore the feasibility and effectiveness of C1 pedicle screw fixation in patients whose atlas vertebral artery groove (defined as the C1 pedicle) height is less than 4 mm, but with a medullary canal. METHODS: From January 2010 to January 2013, 7 patients (6 males, 1 female) with atlantoaxial instability whose C1 pedicle height was less than 4.0 mm on one or both sides were treated by C1 pedicle screw fixation at our institution. Thirteen of the 14 C1 pedicles were less than 4.0 mm in height, but all had a medullary canal. Patients were followed up at regular intervals. Postoperative computed tomography (CT) scans were performed to assess if C1 pedicle screw placement was successful. Clinical outcomes were evaluated according to postoperative complications, the American Spinal Injury Association grading system, and bone graft status. RESULTS: Thirteen C1 pedicles with a height less than 4.0 mm were inserted by 13 3.5- or 4.0-mm-diameter pedicle screws, and one C1 pedicle whose height was 4.1 mm was inserted by a 4.0-mm-diameter pedicle screw. In addition, 14 pedicle screws were inserted in the axis. The mean follow-up period was 23 (range 8-38) months. No neurologic or vascular complications occurred in any of the seven patients. Postoperative CT three-dimensional reconstruction images showed that all 14 pedicle screws were inserted in the C1 pedicles without destruction of the atlas pedicle cortical bone. All patients demonstrated bony fusion 6 months postoperatively. CONCLUSION: If there is a medullary canal in the C1 pedicle, a 3.5- or 4.0-mm-diameter pedicle screw can be safely inserted into the atlas and C1 pedicle screw fixation can be performed without any impact on fixation stability and clinical efficacy, even if the C1 pedicle height is less than 4.0 mm.

The height for screw index (HSI) predicts the development of C2 nerve dysfunction associated with C1 lateral mass screw fixation for atlantoaxial instability.

PURPOSE: To propose a new measure, the height for screw index (HSI), as a predictor of C2 nerve dysfunction in patients who have received posterior C1 lateral mass screw (C1LMS) fixation for atlantoaxial instability and to examine whether the HSI scores correlated with the development of C2 nerve dysfunction through retrospective analysis of 104 C1LMS inserted in 52 patients with atlantoaxial instability. METHODS: The medical records of patients who underwent C1LMS fixation were retrospectively reviewed. C1LMS, 3.5 mm in diameter, was inserted for atlantoaxial stabilization. The sagittal plane of the planned C1LMS trajectory was reconstructed from CT images. The HSI was defined as the difference in height between C2 ganglion and its corresponding foramen. C2 nerve function was assessed using a validated visual analog scale questionnaire. Each foramen receiving C1LMS was considered as a single unit and patients were categorized to group 1, HSI ≥4.0 mm; group 2, HSI <4.0 mm. RESULTS: The mean HSI score was 4.7 ± 0.8 mm (range 3.1-6.5 mm) with 85 (81.7 %) units in group 1, and 19 (18.3 %) units in group 2. Fourteen (13.5 %, 14/104) units developed C2 nerve dysfunction. C2 nerve dysfunction was reported in 4 units in group 1, and 10 units in group 2, respectively. The percentage of C2 nerve dysfunction was significantly higher in group 2 than that in group 1 (P < 0.001, Pearson Chi-square test). CONCLUSIONS: The HSI score correlates with the development of C2 nerve dysfunction in patients receiving C1LMS fixation for atlantoaxial instability and may be a useful predictor of C2 nerve dysfunction.

Targeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motility and invasiveness via a Rho-GTPase- and E-cadherin-dependent mechanism.

microRNAs are small endogenous noncoding RNAs, which post-transcriptionally regulate gene expression. In breast cancer, overexpression of the transmembrane heparan sulfate proteoglycan syndecan-1, a predicted target of the oncomiR miR-10b, correlates with poor clinical outcome. To investigate the potential functional relationship of miR-10b and syndecan-1, MDA-MB-231 and MCF-7 breast cancer cells were transiently transfected with pre-miR-10b, syndecan-1 siRNA or control reagents, respectively. Altered cell behavior was monitored by proliferation, migration and invasion chamber assays, and time-lapse video microscopy. miR-10b overexpression induced post-transcriptional downregulation of syndecan-1, as demonstrated by quantitative real-time PCR (qPCR), flow cytometry, and 3'UTR luciferase assays, resulting in increased cancer cell migration and matrigel invasiveness. Syndecan-1 silencing generated a copy of this phenotype. Adhesion to fibronectin and laminin and basal cell proliferation was increased. Syndecan-1 coimmunoprecipitated with focal adhesion kinase, which showed increased activation upon syndecan-1 depletion. Affymetrix screening and confirmatory qPCR and Western blotting analysis of syndecan-1-deficient cells revealed upregulation of ATF-2, COX-2, cadherin-11, vinculin, actin γ 2, MYL9, transgelin-1, RhoA/C, matrix metalloproteinase 2 (MMP2) and heparanase, and downregulation of AML1/RUNX1, E-cadherin, CLDN1, p21WAF/CIP, cyclin-dependent kinase 6, TLR-4, PAI1/2, Collagen1alpha1, JHDM1D, Mpp4, MMP9, matrilin-2 and ANXA3/A10. Video microscopy demonstrated massively increased Rho kinase-dependent motility of syndecan-1-depleted cells, which displayed increased filopodia formation. We conclude that syndecan-1 is a novel target of the oncomiR miR-10b. Rho-GTPase-dependent modulation of cytoskeletal function and downregulation of E-cadherin expression are identified as relevant effectors of the miR-10b-syndecan-1 axis, which emerges as a promising target for the development of new therapeutic approaches for breast cancer.

Barrierless reactions between two closed-shell molecules. II. Dynamics of F2 + CH3SSCH3 reaction.

A second example of a barrierless reaction between two closed-shell molecules is reported. The reaction F(2)+CH(3)SSCH(3) has been investigated with crossed molecular beam experiments and ab initio calculations. Compared with previous results of the F(2)+CH(3)SCH(3) reaction [J. Chem. Phys. 127, 101101 (2007); J. Chem. Phys. 128, 104317 (2008)], a new product channel leading to CH(3)SF+CH(3)SF is observed to be predominant in the title reaction, whereas the anticipated HF+C(2)H(5)S(2)F channel is not found. In addition, the F+C(2)H(6)S(2)F product channel, the analog to the F+C(2)H(6)SF channel in the F(2)+CH(3)SCH(3) reaction, opens up at collision energies higher than 4.3 kcal/mol. Angular and translational energy distributions of the products are reported and collision energy dependences of the reaction cross section and product branching ratio are shown. The reaction barrier is found to be negligible (<<1 kcal/mol). Multireference ab initio calculations suggest a reaction mechanism involving a short-lived intermediate which can be formed without activation energy.

Barrierless reactions between two closed-shell molecules. I. Dynamics of F(2)+CH(3)SCH(3) reaction.

A detailed experimental and theoretical investigation of the first-reported barrierless reaction between two closed-shell molecules [J. Chem. Phys. 127, 101101 (2007)] is presented. The translational energy and angular distributions of two product channels, HF+CH(2)SFCH(3) and F+CH(3)SFCH(3), determined at several collision energies, have been analyzed to reveal the dynamics of the studied reaction. Detailed analysis of the experimental and computational results supports the proposed reaction mechanism involving a short-lived F-F-S(CH(3))(2) intermediate, which can be formed without any activation energy. Other possible reaction mechanisms have been discriminated. The decay of the intermediate and competition between the two product channels have been discussed.

Dynamics of the F2+CH3SCH3 reaction: a molecule-molecule reaction without entrance barrier.

The F(2)+CH(3)SCH(3) reaction was studied with crossed molecular beam techniques and high level ab initio calculations. Significant reactivity was observed even at low collision energies, consistent with the negligible barrier height obtained from the ab initio calculations. All experimental findings are consistent with a weakly bound reaction intermediate of F-F-S(CH(3))(2) structure, which possesses a special type of three-center four-electron bonding. Analogous intermediates can also explain the reactions of F(2) with CH(3)SH and CH(3)SSCH(3).