LncRNA MIAT facilitated BM-MSCs differentiation into endothelial cells and restored erectile dysfunction via targeting miR-200a in a rat model of erectile dysfunction.

BACKGROUND: Bone-marrow derived mesenchymal stem cells (BM-MSCs) implantation effectively restored rats' erectile dysfunction (ED). Long noncoding RNA (LncRNA)-myocardial infarction-associated transcript (MIAT) has been reported to play an important role in regulating endothelial cells (ECs) function via vascular endothelial growth factor (VEGF) that induced BM-MSCs differentiation into ECs. However, the molecular functions and biological roles of lncRNA MIAT in ED remained unclear. METHODS: The rat model of ED was established. Quantitative real-time PCR (qRT-PCR) and western blotting were used to detect the expression of lncRNA MIAT, von Willebrand factor (vWF), vascular endothelial cadherin (VE-cadherin), endothelial NO synthase (eNOS) and VEGF following BM-MSCs transfection. Erectile function was evaluated by intra-cavernous pressure/mean artery pressure (ICP/MAP). Furthermore, RNA immunoprecipitation (RIP) assay and RNA pull down as well as luciferase reporter assay were carried out to examine the interaction among lncRNA MIAT, miR-200a and VEGF. RESULTS: BM-MSCs restored ED by upregulating lncRNA MIAT. LncRNA MIAT was upregulated in a time-dependent manner during BM-MSCs differentiation into ECs. LncRNA MIAT regulated VEGF via targeting miR-200a, thereby promoting BM-MSCs differentiation into ECs. LncRNA MIAT knockdown in vivo abolished the effect of BM-MSCs on ED. CONCLUSION: LncRNA MIAT promoted BM-MSCs differentiation into ECs and restored ED via miR-200a.

Comparison of intrafascial and non-intrafascial radical prostatectomy for low risk localized prostate cancer.

In this meta-analysis study, we compared the oncological and functional outcomes of intrafascial radical prostatectomy (IFRP) with non-intrafascial radical prostatectomy (NIFRP) in the treatment of patients with low risk localized prostate cancer (PCa). Relevant articles were identified by searching PubMed, EMBASE, Cochrane Library, Ovid, and the ISI Web of Knowledge databases. A total of 2096 patients were included from 7 eligible studies. Results of the pooled data showed that the oncological outcomes including gleason score, positive surgical margin and biochemical free survival rates were similar between the two groups. IFRP was superior to NIFRP with lower postoperative complication rates (RR 0.57, 95% CI 0.38, 0.85, p = 0.006), higher continence rates at 3 months post-operation (RR: 1.14; 95% CI, 1.04, 1.26; p = 0.006), and higher potency rates at 6 months (RR: 1.53; 95% CI, 1.07, 2.18; p = 0.02) and 12 months post-operation (RR: 1.38; 95% CI, 1.11, 1.73; p = 0.005). Additionally, there was a tendency towards higher potency rate in patients ≤65 years old compared with patients >65 years old after IFRP. Overall, these findings suggest that IFRP in young patients with low risk localized PCa had less postoperative complications, shortened time to return to continence and improved potency rate without compromising complete tumor control.

Cavernous nerve reconstruction with autologous vein graft and platelet-derived growth factors.

In this study, we investigated the feasibility of using autologous vein graft and platelet-derived growth factors to bridge transected cavernous nerve in a rat model. A short defect in the bilateral cavernous nerve was created and repaired with vein graft from the right jugular vein or vein graft plus platelet-derived growth factors. The 32 rats were divided into four groups, namely Group 1 - no repair as a negative control, Group 2 - vein graft alone, Group 3 - vein graft plus platelet-derived growth factors, and Group 4 - sham operation as a positive control. We evaluated nerve regeneration and functional recovery using retrograde tracing study with FluoroGold, Toluidine blue staining of cavernous nerve, and the intracavernous pressure at 3 months. Three months after surgery, rich FluoroGold-positive cells were observed in the sham and vein graft plus platelet-derived growth factors group, but very few were found in the no repair group. The number of myelinated axons of regenerated cavernous nerve and intracavernous pressure were increased obviously in the two vein graft groups, especially in the vein graft plus platelet-derived growth factors group. These findings confirm the feasibility of using autologous vein as guides for cavernous nerve regeneration, and the regeneration can be further enhanced when the vein is filled with platelet-derived growth factors.

Antioxidative mechanism of Lycium barbarum polysaccharides promotes repair and regeneration following cavernous nerve injury.

Polysaccharides extracted from Lycium barbarum exhibit antioxidant properties. We hypothesized that these polysaccharides resist oxidative stress-induced neuronal damage following cavernous nerve injury. In this study, rat models were intragastrically administered Lycium barbarum polysaccharides for 2 weeks at 1, 7, and 14 days after cavernous nerve injury. Serum superoxide dismutase and glutathione peroxidase activities significantly increased at 1 and 2 weeks post-injury. Serum malondialdehyde levels decreased at 2 and 4 weeks. At 12 weeks, peak intracavernous pressure, the number of myelinated axons and nicotinamide adenine dinucleotide phosphate-diaphorase-positive nerve fibers, levels of phospho-endothelial nitric oxide synthase protein and 3-nitrotyrosine were higher in rats administered at 1 day post-injury compared with rats administered at 7 and 14 days post-injury. These findings suggest that application of Lycium barbarum polysaccharides following cavernous nerve crush injury effectively promotes nerve regeneration and erectile functional recovery. This neuroregenerative effect was most effective in rats orally administered Lycium barbarum polysaccharides at 1 day after cavernous nerve crush injury.

Role of oxidative stress in surgical cavernous nerve injury in a rat model.

This study investigates the role of oxidative stress in surgical cavernous nerve (CN) injury in a rat model. Eighty-four male Sprague-Dawley rats were randomly divided into three groups: group 1, sham-operated rats; group 2, bilateral CN-crushed rats; and group 3, bilateral CN-transection-and-sutured-immediately rats. Oxidative stress was evaluated by malondialdehyde levels, super oxide dismutase (SOD) activities, and glutathione peroxidase (GPX) activities in serum. Erectile function was assessed by CN electrostimulation at 3 months with mean maximal intracavernous pressure (ICP) and maximal ICP per mean arterial pressure. Nerve injury was assessed by toluidine blue staining of CNs and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining of penile tissue. GPX protein expression and nitrotyrosine-3 (NT-3) levels in penile tissue were measured. Erectile function and the number of myelinated axons of CNs and NADPH-diaphorase-positive nerve fibers were statistically decreased between groups, from sham to crush to transection. For markers, both nerve-injury groups showed increased oxidative stress markers at early time points, with the transection group showing greater oxidative stress than the crushed group and values normalizing to sham levels by week 12. GPX expression and NT-3 levels in penile tissue were in concordance with the results of SOD and GPX. These results show that oxidative stress plays an important role in injured CNs, and different methods of CN injury can lead to different degrees of oxidative stress in a rat model.

[Estradiol stimulates proliferation of prostatic smooth muscle cells via estrogen receptor alpha and IGF1].

OBJECTIVE: To investigate the effects of estrogen receptor alpha (ERa) and insulin-like growth factor 1 (IGF1) on the proliferation of prostatic smooth muscle cells (PSMCs) in vitro. METHODS: The ERalpha shRNA expression frame was subcloned to the pGSadeno adenovirus vector by homologous recombination technology to construct the pGSaaeno-ERalpha vector. After the mouse PSMCs were transfected in vitro by pGSaaeno-ERalpha, the mRNA and protein expression levels of ERalpha were detected by RT-PCR and Western blot respectively. The expression of IGF1 in the ERa-reduced cells was determined by Western blot 6 hours after treatment with 17beta-estradiol (E2) at 10(-8) mol/L. The post-transfection activity of estrogen or exogenous IGF1 in the proliferation of PSMCs was evaluated by MTT chlormetric analysis. RESULTS: After treatment with E2, the proliferation of PSMCs and the expression of the IGF1 gene were significantly increased in the normal control group (P <0.05), but not obviously changed in the ERalpha-siRNA group (P> 0.05). And exogenous IGF1 failed to induce the proliferation of the ERalpha-reduced PSMCs. CONCLUSION: E2 induces the expression of IGF1 via ERalpha, and IGFl, with the interaction of ERalpha, promotes the proliferation of PSMCs.

Estrogen receptor alpha is essential for the proliferation of prostatic smooth muscle cells stimulated by 17ß-estradiol and insulin-like growth factor 1.

Estradiol (E2) and its receptor estrogen receptor alpha (ERα) are implicated in the pathology of stromal-predominant benign prostatic hyperplasia (BPH). Insulin-like growth factor 1(IGF1) is produced primarily by stromal cells in the prostate. Recent study showed that E2-mediated regulation of IGF1 in mouse uterus requires the DNA binding ability of ERα. However, the crosstalk between ERα and IGF1 in the prostate has not been addressed yet. Therefore, in this study we employed mouse prostatic smooth muscle cells (PSMCs) as a model to demonstrate that E2 stimulated the proliferation of PSMCs and up-regulated the expression of IGF1 and its receptor IGF1R as well as cyclin D1 in PSMCs, all of which could be inhibited by shRNA-mediated knockdown of ERα. Furthermore, we found that exogenous IGF1 could not promote cell proliferation and cyclin D1 expression in PSMCs subjected to shRNA-mediated knockdown of ERα. Interestingly, blockage of IGF1R by antibody could inhibit E2-stimulated PSMCs proliferation. Altogether our present study provides the first line of evidence that there is crosstalk between ERα and IGF1 signaling in PSMCs proliferation in which ERα up-regulates the expression of IGF1 and IGF1R, and IGF1 signaling is indispensable to mediate downstream effects of E2 and ERα.

The effect of platelet-rich plasma on cavernous nerve regeneration in a rat model.

The aim of this study was to investigate the effect of platelet-rich plasma (PRP) on cavernous nerve (CN) regeneration and functional status in a nerve-crush rat model. Twenty-four Sprague-Dawley male rats were randomly divided into three equal groups: eight had a sham operation, eight underwent bilateral nerve crushing with no further intervention and eight underwent bilateral nerve crushing with an immediate application of PRP on the site of injury. Erectile function was assessed by CN electrostimulation at 3 months and nerve regeneration was assessed by toluidine blue staining of CN and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining of penile tissue. Three months after surgery, in the group that underwent bilateral nerve crushing with no further intervention, the functional evaluation showed a lower mean maximal intracavernous pressure (ICP) and maximal ICP per mean arterial pressure (MAP) with CN stimulation than those in the sham group. In the group with an immediate application of PRP, the mean maximal ICP and maximal ICP/MAP were significantly higher than those in the injured control group. Histologically, the group with the application of PRP had more myelinated axons of CNs and more NADPH-diaphorase-positive nerve fibres than the injured control group but fewer than the sham group. These results show that the application of PRP to the site of CN-crush injury facilitates nerve regeneration and recovery of erectile function. Our research indicates that clinical application of PRP has potential repairing effect on CN and peripheral nerves.

[Effect of platelet rich plasma on the regeneration of cavernous nerve: experiment with rats].

OBJECTIVE: To investigate the effect of platelet rich plasma (PRP) on the regeneration of injured cavernous nerve (CN). METHODS: Blood was collected from the hearts of 6 SD rats to prepare PRP. 24 male adult rats were randomly divided into 3 equal groups: pure suture group undergoing bilateral CN transaction and pure suture immediately, PRP group undergoing bilateral CN transaction + suture + PRP 200 microl to the site of suture, and sham operation group. 3 months later intracavernous pressure (ICP) was measured by CN electrostimulation and then samples of CN were obtained to undergo pathological examination. RESULTS: 3 months later after surgery, the ICP of the pure suture group was (46 +/- 8) cm H2O, significantly lower than that of the sham group [(109 +/- 13) cm H2O, P < 0.01], and that of the PRP group was (94 +/- 13) cm H2O, significantly higher than that of the pure sutured group (P < 0.01), however, still significantly lower than that of the sham operation group (P < 0.05). The number of axons of CN in the PRP group was (121 +/- 16), significantly higher than that of the pure sutured group (70 +/- 14, P < 0.01); however, still significantly lower than that of the sham operation group (181 +/- 21, P < 0.01). CONCLUSION: PRP can promote the regeneration of injured CN and the recovery of erectile function.

[IFN-gamma and TGF-beta1, levels in the expressed prostatic secretions of patients with chronic abacterial prostatitis].

OBJECTIVE: To investigate the levels of pro-inflammatory cytokine IFN-gamma and anti-inflammatory cytokine TGF-beta1, in the expressed prostatic secretion (EPS) of men with chronic abacterial prostatitis and their clinical significance. METHODS: The levels of IFN-gamma and TGF-beta1, in the EPS of 20 patients with inflammatory chronic pelvic pain syndrome (type III A), 20 patients with non-inflammatory chronic pelvic pain syndrome (type Ill B) and 10 healthy men were measured using enzyme-linked immunosorbent assay (ELISA). The results were analysed comparatively with NIH-chronic prostatitis symptom index (NIH-CPSI). RESULTS: IFN--gamma and TGF-beta1 levels were higher in III ([14.92 +/- 7. 85)], [8477.50 +/- 4612.45] ng/L) and III B ([13.74 +/- 5.96], [7946.50 +/- 5044.06] ng/L) prostatitis patients than those in the controls ([7.47 +/- 1.49], [2462.50 +/- 985.31] ng/L), P < 0.05 and P < 0.001 respectively. There was no statistically significant difference in cytokine levels between III A and Il B prostatitis patients. No correlation was found between NIH-CPSI and cytokine levels, r = 0.02, P = 0.86, r = 0.31, P = 0.76. CONCLUSION: IFN-gamma and TGF-beta1, play a very important role in the etiology of chronic abacterial prostatitis and can be the objective parameters in the diagnosis of chronic abacterial prostatitis.