Liuhua Zhou

In vitro evaluation of the bioactive factors preserved in porcine small intestinal submucosa through cellular biological approaches

The objective of this study was to develop cellular biological approaches to evaluate the potential effect of bioactive factors in porcine small intestinal submucosa (SIS) on bladder regeneration and angiogenesis. For this purpose, we cultured human bladder smooth muscle cell (HBSMC) and human umbilical vein endothelial cell (HUVEC), and then used cellular biological techniques to characterize in vitro biological effect of SIS components on HBSMC and HUVEC. Our results indicated that the SIS components had stimulated the attachment, proliferation, and migration of HBSMC and HUVEC, as well as tube formation by HUVEC on Matrigel. These results implied that the SIS might have preserved a mixture of bioactive factors including cell adhesion factors, mitogenic factors, chemotactic cytokines, and angiogenic factors, and these bioactive factors would have the potential of promoting bladder regeneration and angiogenesis. In conclusion, these cellular biological approaches might be helpful and effective for evaluation of the bioactive factors preserved in porcine SIS before it is used for bladder augmentation in humans.

In Vitro Evaluation of Endothelial Progenitor Cells from Adipose Tissue as Potential Angiogenic Cell Sources for Bladder Angiogenesis

Autologous endothelial progenitor cells (EPCs) might be alternative angiogenic cell sources for vascularization of tissue-engineered bladder, while isolation and culture of EPCs from peripheral blood in adult are usually time-consuming and highly inefficient. Recent evidence has shown that EPCs also exist in the adipose tissue. As adipose tissue is plentiful in the human body and can be easily harvested through a minimally invasive method, the aim of this study was to culture and characterize EPCs from adipose tissue (ADEPCs) and investigate their potential for the neovascularization of tissue-engineered bladder. Adipose stromal vascular fraction (SVF) was isolated and used for the culture of ADEPCs and adipose derived stem cells (ADSCs). After SVF was cultured for one week, ADEPCs with typical cobblestone morphology emerged and could be isolated from ADSCs according to their different responses to trypsinization. Rat bladder smooth muscle cells (RBSMCs) were isolated and cultured from rat bladder. RBSMCs exhibited typical spindle-shaped morphology. ADEPCs had higher proliferative potential than ADSCs and RBSMCs. ADEPCs stained positive for CD34, Stro-1, VEGFR-2, eNOS and CD31 but negative for α-SMA, CD14 and CD45. ADSCs stained positive for CD34, Stro-1 and α-SMA but negative for VEGFR-2, eNOS, CD31, CD14 and CD45. RBSMCs stained only positive for α-SMA. ADEPCs could be expanded from a single cell at an early passage to a cell cluster containing more than 10,000 cells. ADEPCs were able to uptake DiI-Ac-LDL, bind UEA-1 and form capillary-like structures in three-dimensional scaffolds (Matrigel and bladder acellular matrix). ADEPCs were also able to enhance the human umbilical vein endothelial cells’ capability of capillary-like tube formation on Matrigel. Additionally, significantly higher levels of mRNA and protein of vascular endothelial growth factor were found in ADEPCs than in RBSMCs. These results suggest the potential use of ADEPCs as angiogenic cell sources for engineering bladder tissue.

Efficacy and safety of local anaesthetics for premature ejaculation: a systematic review and meta-analysis

To assess the efficacy and safety of local anaesthetics for premature ejaculation (PE), a systematic review of the literature was performed using the Cochrane Library, PUBMED and EMBASE. We screened and retrieved the randomized controlled trials on the treatment of PE with local anaesthetics. End points included intravaginal ejaculation latency time (IELT), patient-reported outcome assessments and adverse events. Meta-analyses were conducted with Stata 11.0. In total, seven publications involving 566 patients with local anaesthetics and 388 with placebos strictly met our eligibility criteria. Meta-analyses showed that after the patients were treated with the local anaesthetics, the value of the standardized mean difference of the changes in IELT was 5.02 (95% CI: 3.03-7.00). A higher rate of adverse events occurred compared with placebos (odds ratio: 3.30, 95% CI: 1.71-6.36), but these events were restricted to local side effects. In addition, significantly greater improvement was observed in patient-reported outcomes. In summary, local anaesthetics can prolong IELT and improve ejaculatory control and sexual satisfaction.

In vitro comparative evaluation of recombinant growth factors for tissue engineering of bladder in patients with neurogenic bladder.

BACKGROUND To compare the effects of various recombinant growth factors on bladder regeneration and angiogenesis for tissue engineering of bladder in patients with neurogenic bladder through in vitro cellular biological methods. MATERIALS AND METHODS Human bladder smooth muscle cells (HBSMCs) and human bladder urothelial cells (HBUCs) were cultured from patients with neurogenic bladder and used for comparative evaluations of various growth factors. Human umbilical vein endothelial cells (HUVECs) were also used. Eight potential growth factors, platelet-derived growth factor BB (PDGF-BB), platelet-derived growth factor CC (PDGF-CC), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), insulin-like growth factor 1 (IGF-1), hepatocyte growth factor (HGF), and transforming growth factor beta 1 (TGF-β1), were selected and their effects on the proliferation, migration, and wound healing of HBSMCs, HBUCs, and HUVECs were compared. RESULTS PDGF-BB, PDGF-CC, bFGF, VEGF, IGF-1, or HGF enhanced the proliferation, migration, and wound healing of HBSMCs, whereas TGF-β1 inhibited their proliferation. Proliferation, migration, and wound healing of HBUCs and HUVECs were enhanced by bFGF, VEGF, EGF, IGF-1, or HGF, whereas inhibited by TGF-β1. PDGF-BB failed to enhance cell activity of HUVECs, whereas PDGF-CC could enhance their migration and wound healing. PDGF-BB, EGF, and VEGF were the most potent factors for stimulating the activities of HBSMCs, HBUCs, and HUVECs, respectively. CONCLUSIONS Our findings suggest the potential use of a combination of PDGF-BB, EGF, and VEGF for bladder regeneration and angiogenesis. The synergetic effects of the three growth factors on cell activities in a three-dimensional scaffold and an animal model with neurogenic bladder need to be further evaluated.

Clinical characteristics and penile afferent neuronal function in patients with primary delayed ejaculation

Primary delayed ejaculation (DE) is a relatively uncommon condition and has not been studied broadly. In this study, we aimed to investigate the clinical characteristics and penile afferent neuronal function using somatosensory evoked potentials in patients with primary DE. Twenty‐four patients with primary DE and 24 age‐matched normally potent men were enrolled in this study. Results indicated that patients with primary DE had remarkably higher frequency of masturbatory activity (especially, some with idiosyncratic styles), lower night emissions, longer intravaginal ejaculation latency time (IELT), higher anxiety and depression states (p = 0.010, p = 0.017, p < 0.001, p < 0.001, p < 0.001 respectively). In addition, the mean penile shaft sensory threshold values in the patients were considerably higher than those in the healthy men (p < 0.001). Mean latencies of dorsal nerve somatosensory evoked potential DNSEP were 4.32 ms longer in the DE group than those in the control group (p < 0.001). However, the latencies of glans penis somatosensory evoked potential (GPSEP) between the two group showed no significant difference (p = 0.985). At the same time, in comparison with the control group, the amplitudes of DNSEP were considerably lower in the DE group (p = 0.016), but not in the amplitudes of GPSEP (p = 0.934). This study indicates that the patients with primary DE appear to have penile shaft rather than glans hyposensitivity and hypoexcitability, and adaptation to a certain masturbatory technique (higher and idiosyncratic) may be related to the causes of primary DE, which is also associated with lower night emissions, longer IELT, higher anxiety and depression states.

The Sympathetic Skin Response Located in the Penis as a Predictor of the Response to Sertraline Treatment in Patients with Primary Premature Ejaculation

INTRODUCTION The pathologic mechanisms of primary premature ejaculation (PPE) are complex and multifactorial, and hyperactivity of the sympathetic nervous system is one of the mechanisms. AIM To examine the effects of sertraline on sympathetic nervous system activity and assess the predictive value of the sympathetic skin response located in the penis (PSSR) on the response to sertraline treatment in PPE patients. METHODS Sixty-one patients with PPE were recruited. Each received 50 mg sertraline daily for 8 weeks. Before and after the experiment, the patients were evaluated for PSSR tests and sexual performance parameters. Additionally, based on the latency of PSSR, we divided the patients into a normal PSSR group and an abnormal PSSR group, and compared the sertraline treatment efficacy between the two groups. MAIN OUTCOME MEASURES Changes in intravaginal ejaculation latency time (IELT) and the Chinese premature ejaculation index-5 (CIPE-5), and the latencies and amplitudes of PSSR after sertraline treatment. RESULTS Overall, 58 (95.1%) patients completed the entire study and were analyzed. After the 8-week sertraline treatment, compared with those of pretreatment, IELT and CIPE-5 scores were significantly increased (both P < 0.001), and the amplitudes and latencies of PSSR in the PPE patients were remarkably decreased and prolonged, respectively (both P < 0.001). In addition, the changes of the latencies of PSSR were positively correlated with the increment of IELT (r = 0.375, P = 0.004). The treatment outcome was better in patients with a baseline abnormal PSSR than in those with a baseline normal PSSR (P = 0.021). CONCLUSIONS These results suggest that clinical improvement in response to sertraline in the PPE patients, at least in part, is mediated through reducing sympathetic nervous system activity indexed by PSSR. Measurement of the PSSR appears to provide useful information for predicting treatment responses in the PPE patients.

Stem cells in a tissue-engineered human airway

www.thelancet.com Vol 379 April 21, 2012 1487 We agree with Jinichi Mori and colleagues that it would be interesting to determine what subpopulation of young patients with diff use large B-cell lymphoma, if any, would benefi t preferentially from intensifi ed immunochemotherapy with R-ACVBP. As stated in our paper, we did not identify any clinical or biological factor to suggest the existence of such a category of patients. Studies to investigate tumour immunophenotype and gene expression profi le in patients whose samples are available are ongoing. Since R-ACVBP is a frontline treatment regimen, it could not be assigned according to the results of interim PET. However, interim PET is currently being explored by our group to help drive the decision about consolidative high-dose chemotherapy and stem-cell transplantation after R-ACVBP induction in patients with high-risk diff use large B-cell lymphoma.

AB110. In vitro evaluation of endothelial progenitor cells from adipose tissue as potential angiogenic cell sources for bladder angiogenesis

Objective Autologous endothelial progenitor cells (EPCs) might be alternative angiogenic cell sources for vascularization of tissue-engineered bladder, while isolation and culture of EPCs from peripheral blood in adult are usually time-consuming and highly inefficient. Recent evidence has shown that EPCs also exist in the adipose tissue. As adipose tissue is plentiful in the human body and can be easily harvested through a minimally invasive method, the aim of this study was to culture and characterize endothelial progenitor cells from adipose tissue (ADEPCs) and investigate their potential for the neovascularization of tissue-engineered bladder. Methods Adipose stromal vascular fraction (SVF) was isolated and used for the culture of ADEPCs and adipose derived stem cells (ADSCs). Rat bladder smooth muscle cells (RBSMCs) were isolated and cultured from rat bladder. Cells were characterized by immunofluorescence staining and flow cytometric analysis. The capabilities of cell proliferation, DiI-Ac-LDL uptake, and UEA-1 binding were evaluated. Real-time RT-PCR and ELISA were performed to assess the expression of angiogenic factors. ADEPCs were seeded onto matrigel and bladder acellular matrix (BAM), and the capability of tube formation in these scaffolds was evaluated. Results After SVF was cultured for one week, ADEPCs with typical cobblestone morphology emerged and could be isolated from ADSCs according to their different responses to trypsinization. RBSMCs exhibited typical spindle-shaped morphology. ADEPCs had higher proliferative potential than ADSCs and RBSMCs. ADEPCs stained positive for CD34, Stro-1, VEGFR-2, eNOS and CD31 but negative for α-SMA, CD14 and CD45. ADSCs stained positive for CD34, Stro-1 and α-SMA but negative for VEGFR-2, eNOS, CD31, CD14 and CD45. RBSMCs stained only positive for α-SMA. ADEPCs could be expanded from a single cell at an early passage to a cell cluster containing more than 10,000 cells. ADEPCs were able to uptake DiI-Ac-LDL, bind UEA-1 and form capillary-like structures in three-dimensional scaffolds (matrigel and bladder acellular matrix). ADEPCs were also able to enhance the human umbilical vein endothelial cells’ capability of capillary-like tube formation on matrigel. Additionally, significantly higher levels of mRNA and protein of vascular endothelial growth factor were found in ADEPCs than in RBSMCs. Conclusions These results suggest the potential use of ADEPCs as angiogenic cell sources for engineering bladder tissue.