Penile Tissue Research Articles

Caveolin-1 scaffolding domain peptide prevents corpus cavernosum fibrosis and erectile dysfunction in bilateral cavernous nerve injury-induced rats.

Corpus cavernosum (CC) fibrosis significantly contributes to post-radical prostatectomy erectile dysfunction (pRP-ED). Caveolin-1 scaffolding domain (CSD)-derived peptide has gained significant concern as a potent antagonist of tissue fibrosis. However, applying CSD peptide on bilateral cavernous nerve injury (BCNI)-induced rats remains uninvestigated. The aim was to explore the therapeutic outcome and underlying mechanism of CSD peptide for preventing ED in BCNI rats according to the hypothesis that CSD peptide may exert beneficial effects on erectile tissue and function following BCNI through limiting collagen synthesis in CC smooth muscle cells (CCSMCs) and CC fibrosis. After completing a random assignment of male Sprague Dawley rats (10 weeks of age), BCNI rats received either saline or CSD peptide treatment, as opposed to sham-operated rats. The evaluations of erectile function (EF) and succedent collection and histological and molecular biological examinations of penile tissue were accomplished 3weeks postoperatively. In addition, the fibrotic model of CCSMCs was used to further explore the mechanism of CSD peptide action in vitro. The assessments of EF, SMC/collagen ratio, α-smooth muscle actin, caveolin-1 (CAV1), and profibrotic indicators expressions were conducted. BCNI rats exhibited significant decreases in EF, SMC/collagen ratio, α-SMA, and CAV1 levels, and increases in collagen content together with transforming growth factor (TGF)-β1/Smad2 activity. However, impaired EF, activated CC fibrosis, and Smad2 signaling were attenuated after 3weeks of CSD peptide treatment in BCNI rats. In vitro, TGF-β1-induced CCSMCs underwent fibrogenetic transformation characterized by lower expression of CAV1, higher collagen composition, and phosphorylation of Smad2; then, the delivery of CSD peptide could significantly block CCSMC fibrosis by inactivating Smad2 signaling. Based on available evidence of CSD peptide in the prevention of ED in BCNI rats, this study can aid in the development and clinical application of CSD peptide targeting pRP-ED. This study provides data to suggest that CSD peptide protects against BCNI-induced deleterious alterations in EF and CC tissues. However, the available evidence still does not fully clarify the detailed mechanism of action of CSD peptide. Administration of CSD peptide significantly retarded collagen synthesis in CCSMCs, limited CC fibrosis, and prevented ED via confrontation of TGF-β1/Smad signaling in BCNI rats.

Extract of Pfaffia glomerata Ameliorates Paroxetine-Induced Sexual Dysfunction in Male Mice and the Characterization of Its Phytoconstituents by UPLC-MS.

Pfaffia glomerata extract (PGE) has a variety of biological activities. However, its ameliorative effect on and exact working mechanism in male sexual dysfunction are still poorly understood. This study aims to evaluate the ameliorative effect of PGE on paroxetine (PRX)-induced sexual dysfunction in male mice and uses molecular docking technology to investigate its underlying mechanism. In this work, PRX-induced sexual dysfunction was caused and PGE was gavaged in mice for 28 days. The results show that PGE significantly improved the sexual performance of mice and reduced the damage to testicular tissues. Further studies showed that PGE restored serum sex hormones to normal levels and increased nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) levels as well as nitric oxide synthase (NOS) activity in penile tissues, while also decreasing phosphodiesterase-5 (PDE-5) activity, thereby maintaining normal penile erection in mice. In addition, PGE improved the activities of enzymes (LDH, ACP, and ALP) related to energy metabolism in the testis and significantly increased sperm count and viability in mice. Furthermore, the molecular docking results show that all eight compounds in PGE could form a stable complex with PDE-5 and inhibit the activity of PDE-5. In conclusion, PGE had an ameliorative effect on PRX-induced sexual dysfunction, suggesting that PGE has a potential protective effect on male sexual health.

Radiofrequency energy in the treatment of erectile dysfunction-a novel cohort pilot study on safety, applicability, and short-term efficacy.

The erectile mechanism depends, in part on the intactness of the collagen components in the penis. As such, impaired collagen may have a deleterious effect on erectile function. Radiofrequency energy has been shown to renew and restore spatial structural arrangement of collagen fibers; therefore, treatment of erectile dysfunction with radiofrequency could lead to anatomical and physiological changes at the penile tissue level and could lead to improvement in the erectile mechanism. We conducted this study to assess the effect of radiofrequency treatment on erection quality. We evaluated the safety, applicability, and efficacy of a self-applied, handheld, low-intensity radiofrequency device (Vertica®) in men with moderate and mild-to-moderate organic erectile dysfunction. The treatment protocol consisted of 12 treatments (twice a week during the 1st month, and once a week during the 2nd month), and each participant treated himself individually. Treatment outcomes were evaluated using the International Index of Erectile Function, Erection Hardness Scale, Erectile Dysfunction Index of Treatment Satisfaction, Benefit, Satisfaction & Willingness to continue, Quality of Erection Questionnaire, Sexual Quality of Life questionnaires and specific questions addressing side effects and ease of use. Twenty-eight out of 32 men (mean age 59.5 ± 9.8, range: 41-78 years) completed a one-month follow-up after treatment. Mean International Index of Erectile Function (43.7. ± 7.8 vs. 60.9 ± 10.8, p < 0.01), International Index of Erectile Function -Erection Function domain (16.8 ± 3.1 vs. 24.4 ± 4.4, p < 0.001), and Erection Hardness Scores (2.2 ± 0.8 vs. 3.2 ± 0.5, p = 0.01) were all significantly improved. Fifty percent of patients achieved normal erectile function parameters according to the International Index of Erectile Function -Erection Function domain score >25. High mean scores were achieved in the Erectile Dysfunction Index of Treatment Satisfaction (76.8 ± 20.3), Benefit, Satisfaction & Willingness to continue (4.83 ± 1.1), Quality of Erection Questionnaire (73.4 ± 23.8), and Sexual Quality of Life (67 ± 29.4) questionnaires. No side effects were reported and participants rated the device as very comfortable, simple, and easy to operate.

Evaluation of androgen receptor markers in erectile dysfunction.

Steroid hormones, such as testosterone, play a crucial role in modulating the development of male internal and external genitalia as well as secondary sex characteristics by binding to the androgen receptor. Once bound, androgen receptor operates as an inducible transcription factor, interacting with a multitude of co-regulators to initiate various downstream signaling pathways. The androgen saturation hypothesis posits that beyond a specific threshold, androgen receptor binding and functionality remain unaltered despite an increase in serum testosterone levels. The objective of this study was to explore the expression of these proteins in penile tissue samples from men with severe erectile dysfunction to enhance our understanding of the influence of serum testosterone on androgen receptor function. Patients undergoing surgical management for high-grade ED at our institution were invited to participate in the study. During inflatable penile prosthesis surgery, corpus cavernosum biopsy was obtained. Protein was extracted from each sample for western blot analysis which was probed with androgen receptor, heme oxygenase, inducible nitric oxide synthase, and phosphodiesterase type 5 antibodies with GAPDH for protein normalization. 12 men agreed to participate in this study. Serum testosterone levels were obtained from all participants on the morning of their surgery. The median testosterone level was 300.15 ng/dL. Our findings revealed a decrease in androgen receptor and inducible nitric oxide synthase expression at serum testosterone levels below 300 ng/dL (p = 0.022, 0.03). Similarly, hemeoxygenase and phosphodiesterase type 5 expression levels were significantly lower at serum T concentrations below 200 ng/dL (p = 0.017, 0.014). These data showed a significant decrease in the expression of proteins downstream of the androgen receptor at lower serum T levels. This suggests a potential correlation between serum T concentration and androgen receptor signaling and supports a potential saturation value between 200 and 300 ng/dL.

FAP-targeted CAR T-cell therapy: A promising approach for the treatment of Peyronie’s disease

Peyronie’s disease is a fibro-inflammatory disease of penile connective tissue. Fibrotic plaque formed in Peyronie's disease can cause penile curvature disorders, anxiety disorders, and disruption of sexual life. Medical and surgical regimens are available to treat Peyronie's disease. However, current pharmacological treatment modalities are ineffective and may not prevent recurrences. Tissue losses and recurrences may occur in Peyronie's surgery performed for treatment. This situation creates the need for new-generation therapeutic agents. This article hypothesized how adoptive cell therapy with CAR T-cells, a popular and effective methodology, can be used to treat Peyronie's disease. Our hypothesis is centered around the development of a T cell equipped with a chimeric antigen receptor specifically designed to target FAP, which serves as an indicator of activated fibroblasts. We think it will be a promising therapy model for Peyronie's disease in the future because adoptive cell therapy can prevent recurrences because it takes place in the body for a long time and can reduce plaque before fibrotic tissue grows.

Effects of endoplasmic reticulum stress on erectile function in rats with cavernous nerve injury.

Erectile dysfunction (ED) occurs in an increasing number of patients after radical prostatectomy and cystectomy, and the phenotypic modulation of corpus cavernosum smooth muscle cells is closely related to ED. To determine whether endoplasmic reticulum stress (ERS) is implicated in the phenotypic modulation of ED induced by bilateral cavernous nerve injury (BCNI). In total, 36 Sprague-Dawley rats were randomly divided into 3 groups: sham, in which rats received sham surgery with bilateral cavernous nerve exposure plus phosphate-buffered saline; control, in which rats received BCNI plus phosphate-buffered saline; and experimental, in which rats received BCNI plus 4-phenylbutyric acid. Analysis of variance and a Bonferroni multiple-comparison test were utilized to evaluate differences among groups. Erectile function, smooth muscle/collagen ratios, and the expression levels of phenotypic modulation and ERS were measured. Two ratios-maximum intracavernosal pressure/mean arterial pressure and smooth muscle/collagen-were decreased in the control group as compared with the sham group. In penile tissue, there was increased expression of GRP78 (78-kDa glucose-regulated protein), p-PERK/PERK (phosphorylated protein kinase R-like endoplasmic reticulum kinase/protein kinase R-like endoplasmic reticulum kinase), caspase 3, CHOP (C/EBP homologous protein), and OPN (osteopontin) but decreased expression of nNOS (neuronal nitric oxide synthase) and α-SMA (α-smooth muscle actin). As compared with the control group, erectile function was improved and pathologic changes were partially recovered in the experimental group. The present study demonstrated that ERS is involved in ED caused by cavernous nerve injury, thereby providing a new target and theoretical basis for clinical treatment. The present study demonstrated for the first time that ERS is related to ED caused by cavernous nerve injury. Inhibition of ERS reverses phenotypic modulation and improves erectile function in rats with BCNI. Additional in vitro studies should be performed to verify these conclusions and explore the specific mechanism of phenotypic modulation. The present study demonstrated that inhibiting ERS reverses phenotypic modulation and enhances erectile function in rats with BCNI.

Low androgen levels induce ferroptosis of rat penile cavernous endothelial cells.

Endothelial dysfunction caused by low androgen levels in penile tissue can lead to erectile dysfunction. The exact mechanism of endothelial dysfunction has not been thoroughly studied. The study sought to verify whether low androgen levels induce ferroptosis of endothelial cells in rat penile tissue. Rat penile cavernous endothelial cells (CP-R133) were divided into a no-androgen group (Dihydrotestosterone (DHT): 0nmol/L), very low-androgen group (DHT: 0.1nmol/L), low-androgen group (DHT: 1nmol/L), DHT = 10nmol/L group, DHT (0nmol/L) + ferrostatin-1 (Fer-1) group, DHT (0.1nmol/L) + Fer-1 group, DHT (1nmol/L) + Fer-1 group, DHT (10nmol/L) + Fer-1 group. Cell viability, intracellular ferrous ion (Fe2+), malondialdehyde (MDA), GSH into oxidized glutathione (GSSG), reactive oxygen species (ROS), nitric oxide (NO), transferrin receptor 1 protein (TfR1), solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4), endothelial nitric oxide synthase (eNOS), and phospho-eNOS (p-eNOS) were detected. Low androgen levels could induce ferroptosis of rat penile cavernous endothelial cells in vivo by upregulating the expressions of TfR1 and ACSL4 and downregulating the expressions of SLC7A11 and GPX4. Cell viability, the levels of glutathione (GSH), NO, SLC7A11, GPX4, and p-eNOS/eNOS in the DHT = 0nmol/L group were lower than those in the other groups (P < .05). The levels of Fe2+, ROS, MDA, GSSG, TfR1, and ACSL4 in the DHT = 0nmol/L group were higher than those in the other groups (P < .05). Cell viability and the levels of GSH, NO, SLC7A11, GPX4, and p-eNOS/eNOS in the DHT = 1nmol/L group were lower than those in the DHT (1nmol/L) + Fer-1 group, DHT = 10nmol/L group, and DHT (10nmol/L) + Fer-1 group (P < .05). The levels of Fe2+, ROS, MDA, GSSG, TfR1, and ACSL4 in the DHT = 1nmol/L group were higher than those in the DHT (1nmol/L) + Fer-1 group, DHT = 10nmol/L group, and DHT (10nmol/L) + Fer-1 group (P < .05). A ferroptosis inhibitor might be a novel drug for treating erectile dysfunction caused by low androgen level. The results of this study need to be further confirmed in in vitro and in human studies. Meanwhile, further investigation is needed to clarify whether low androgen levels affect ferroptosis of rat penile cavernous smooth muscle and nerve cells. Low androgen levels can induce ferroptosis of endothelial cells in rat penile tissue. Inhibition of ferroptosis can reverse endothelial dysfunction caused by low androgen levels.

Traditional Chinese Medicine formula Wubi Shanyao Pills protects against reproductive aging by activating SIRT1/3 to reduce apoptosis

Ethnopharmacological relevanceThe study of male reproductive aging and its associated concerns holds significant importance within the realm of health issues affecting the elderly population. Wubi Shanyao Pills (WSP), a traditional Chinese patent medicine originating from the Tang Dynasty, has been recognized for its ability to enhance male sexual functions while also tonifying the kidney and spleen. Nevertheless, the precise effects and underlying mechanisms through which WSP ameliorates the decline in reproductive function among aging men remain uncertain. Aim of the studyThis study elucidated the distinctive impacts of WSP on ameliorating the decline in reproductive function caused by natural aging, as well as its underlying mechanisms. Materials and methodsInitially, male mice at the age of 15 months were administered WSP orally at doses of 0.375, 0.75, and 1.50 g/kg per day for a duration of 8 consecutive weeks. The impact of WSP on age-related manifestations in naturally aging mice was assessed based on their behavioral performance. The renal function of the mice was evaluated by measuring serum biochemical indicators, including Creatinine (CR), Uric acid (UA), and Blood urea nitrogen (BUN). Additionally, Superoxide dismutase (SOD) and Malonaldehyde (MDA) levels in renal tissue were determined using applied chemistry methods. Then assessed the levels of Nitric oxide (NO), Total nitric oxide synthase (T-NOS), Guanosine cyclase (GC), and Cyclic guanosine monophosphate (cGMP) in the penile tissue, as well as the expression of Endothelial nitric oxide synthase (eNOS) and Guanylate Cyclase Activator (GUCA) protein, in order to investigate the erectile function of the penis. Additionally, the quality of epididymal sperm was examined using an electron microscope. Furthermore, the serum sex hormone level and related protein expression were determined through the utilization of enzyme-linked immunosorbent assay and immunohistochemistry techniques. Pathological alterations and the ultrastructure of the testis were investigated using hematoxylin-eosin staining and transmission electron microscopy. Subsequently, the apoptosis of spermatogenic cells in the testes was assessed employing TUNEL, immunofluorescence, western blotting, and quantitative real-time polymerase chain reaction. ResultsThe administration of WSP has been found to enhance the behavioral performance and sexual behavior in aged mice. It's also could increase in serum levels of CR, UA, and BUN, as well as the elevation of SOD activity in kidney tissue, which subsequently leads to a reduction in MDA levels and an improvement in the structural damage caused by aging in the kidney tissue. Consequently, the renal function is enhanced. Additionally, WSP has been observed to elevate the levels of NO, T-NOS, GC, and cGMP in penile tissue, along with an increase in eNOS and GUCA protein expression, indicating an improvement in penile erectile function. The administration of WSP resulted in a decrease in the occurrence of programmed cell death in testicular germ cells, leading to an enhancement in sperm quality and the overall function of testicular spermatogenesis. This improvement can be attributed to the modulation of hormone levels and the regulation of SIRT1/3, p53, FOXO3, Bax, and Caspase-3 expression. ConclusionCollectively, our findings indicate that the administration of WSP has the potential to impede the occurrence of programmed cell death in testicular cells by modulating the expression of SIRT1/3 and subsequent genes associated with apoptosis. Consequently, this regulatory mechanism facilitates the proliferation of testicular cells and sustains the spermatogenic function of the testes. Consequently, by modulating the levels of sexual hormones in naturally aging mice, WSP ultimately enhances the quality of sperm and reproductive function. Concurrently, it also ameliorates age-related behavioral changes, renal function, and erectile function.

Management of penile post-circumcision ischemia by pentoxifylline infusion and hyperbaric oxygen therapy

BackgroundPost-circumcision penile ischemia is a devastating complication. We will present our experience in managing children with various forms of penile ischemia.Materials and methodsThis cohort prospective observational and interventional study was performed on all male children with post-circumcision penile ischemia between April 2017 and October 2021. A designed and approved protocol includes a combination of early pentoxifylline infusion, hyperbaric oxygen inhalation, early catheterization, and appropriate surgical debridement were applied for patients with deep ischemia 11/23, mainly the necrotic skin and subcutaneous tissues. Data of patient age, anesthesia method, monopolar diathermy usage, early presentation and positive wound culture were collected and analyzed statistically.ResultsDuring the study period 3,382 children were circumcised for non-medical reasons; 23 children were diagnosed with penile ischemia (0.7%), among other complications (9%). Most of the penile ischemia is associated with the use of monopolar diathermy (74%). The use of compressive wound dressing to control post-circumcision bleeding and infections is also responsible for ischemia in 52.2% and 43.5% of the cases. Inexperienced physicians were commonly responsible for ischemia (73.9%). Patients managed at first 24 h had better outcomes than those who were presented later (p = 0.001).ConclusionIn children with post-circumcision penile ischemia, a combination of hyperbaric oxygen therapy and pentoxifylline is especially effective for patients with skin and facial necrosis, this management reduces penile tissue loss.

Bilateral Technique of Penis Flap with Scrotum Skin in Penile Paraffinoma Cases: A Case Report

Background: Paraffin injection into the penis is one way that people see it as increasing penis size. But many do not know that paraffin injection actually causes side effects in the form of penile paraffinoma. This study aimed to describe the management of paraffinoma using bilateral techniques.&#x0D; Case presentation: A 40-year-old married man came to the emergency room at Samuel J. Moeda Naval Hospital, Kupang, with complaints of pain and swelling of the penis for 6 hours. The pain felt to get worse when the penis was erected. The genital organs found swelling of the entire penile skin accompanied by tenderness and induration. The bilateral scrotal flap technique was because the scrotal skin had many blood vessels, good flexibility, little adipose tissue, thin skin, and could feel erogenous sensations almost the same as the physiological characteristics of the penile skin.&#x0D; Conclusion: Bilateral scrotal flap technique on the penile shaft is a surgical technique that is quite effective in eradicating paraffin from penile tissue.

Almond-citrus peel enriched short bread modulates sexual behaviour and enzymes linked with erectle dysfunction in hypertensive rats

Almond nut (Terminalia catappa) and orange peel (Citrus sinensis) are commonly used as nutraceuticals in folklore for the treatment of degenerative diseases. Erectile dysfunction is a pathology with various predisposing factors such as hypertension and hypercholesterolemia. This study aim to evaluate the effect of almond-citrus peel enriched shortbread on sexual function and biochemical parameters linked with erectile dysfunction in high fat diet (HFD)/Nω-nitro-l-arginine methyl ester hydrochloride (l-NAME)- induced male rats. The experimental rats were distributed into eight groups and fed with shortbread enriched with varying inclusions of almond and citrus peel. Sexual behavioral assessments (mounting number, mounting latency, intromission number and intromission latency) were evaluated. Thereafter, the activities of phosphodiesterase-5 (PDE-5),arginase, malondialdehyde (MDA), reactive oxygen species (ROS), antioxidant indices and nitric oxide (NO) level were evaluated. The result showed significant elevation in mounting latency, intromission latency, MDA level as well as arginase and PDE-5 activities in the penile tissue of the untreated rats. Conversely, the mounting number, intromission number, antioxidant status and NO level were significantly decreased in the untreated groups. Notably, the treatment with almond-citrus peel enriched shortbread and the individual effect of almond (50%) and citrus peel (0.2%) all reversed these trends in the hypertensive rats. However, treatment with the almond (50%)-citrus peel (0.2%) shortbread reinstated the altered sexual behaviour in male rats, enhanced antioxidant status and suppressedphosphodiesterase-5 and arginase activities in HFD/l-NAME-induced rats. The results suggest that almond and citrus peel offer potentials as therapeutic agent in the prevention and management of erectile dysfunction associated with hypertension. Intriguingly, the almond (50%)-citrus (0.2%) formulated shortbread had the best antioxidative and erectogenic effects.However, further in vivo experiments and clinical trials are recommended.

War-related urethral and penile injuries' management and short-term outcomes: Experience from war in Yemen

Abstract Objectives To investigate the mechanisms of injury, wound patterns, surgical management, and outcomes of urethral and penile injuries caused by firearms and explosives during the ongoing war in Yemen. Materials and methods The present prospective descriptive study included 46 patients who sustained penile and/or urethral injuries during the ongoing Yemeni war. The patients received primary surgical care in Yemen and were referred for definitive treatment in Egypt between October 2018 and October 2020. Patient demographics, cause of injury, site and severity of injury, and associated injuries were evaluated together with management outcomes. Results The median age was 28 years (range, 18–50 years); 27 patients (58.7%) had posterior urethral injuries, 11 (23.9%) of whom had posterior urethral distraction defect injuries, 5 (10.9%) had a posterior urethral distraction defect injuries associated with urethrorectal fistulas, and the remaining 11 patients developed posterior urethral strictures; 9 patients (19.6%) sustained anterior urethral injuries, and 10 (21.7%) had isolated penile tissue injuries. Injuries to the penis (90%) and anterior urethra (67%) were primarily due to improvised explosive devices, whereas injuries to the posterior urethra (85%) were mostly caused by firearms. During the posttreatment follow-up period (6–12 months), 4 patients (8.7%) developed recurrent urethral strictures. Conclusions Urethral injuries caused by firearms and improvised explosive devices showed a greater association with urethrorectal fistulas or penile tissue injuries than what is typically observed in our practice. Management of these injuries by highly experienced urologists enhances patient outcomes and minimizes future complications.

BMP2 restores erectile dysfunction through neurovascular regeneration and fibrosis reduction in diabetic mice.

The odds of erectile dysfunction are three times more prevalent in diabetes. Severe peripheral vascular and neural damage in diabetic patients responds poorly to phosphodiesterase-5 (PDE5) inhibitors. However, bone morphogenetic protein 2 is known to be involved in angiogenesis. To assess the efficacy of bone morphogenetic protein 2 in stimulating angiogenesis and augmenting nerve regeneration in a mouse model of diabetic-induced erectile dysfunction. The induction of diabetes mellitus was performed by streptozotocin (50mg/kg daily) administered intraperitoneally for 5 successive days to male C57BL/6 mice that were 8weeks old. Eight weeks post-inductions, animals were allocated to one of five groups: a control group, a streptozotocin-induced diabetic mouse group receiving two intracavernous 20μL phosphate-buffered saline injections, or one of three bone morphogenetic protein 2 groups administered two injections of bone morphogenetic protein 2 protein (1, 5, or 10μg) diluted in 20μL of phosphate-buffered saline within a 3-day interval between the first and second injections. The erectile functions were assessed 2 weeks after phosphate-buffered saline or bone morphogenetic protein 2 protein injections by recording the intracavernous pressure through cavernous nerve electrical stimulation. Angiogenic activities and nerve regenerating effects of bone morphogenetic protein 2 were determined in penile tissues, aorta, vena cava, the main pelvic ganglions, the dorsal roots, and from the primary cultured mouse cavernous endothelial cells. Moreover, fibrosis-related factor protein expressions were evaluated by western blotting. Erectile function recovery to 81% of the control value in diabetic mice was found with intracavernous bone morphogenetic protein 2 injection (5μg/20μL). Pericytes and endothelial cells were extensively restored. It was confirmed that angiogenesis was promoted in the corpus cavernosum of diabetic mice treated with bone morphogenetic protein 2 through increased ex vivo sprouting of aortic rings, vena cava and penile tissues, and migration and tube formation of mouse cavernous endothelial cells. Bone morphogenetic protein 2 protein enhanced cell proliferation and reduced apoptosis in mouse cavernous endothelial cells and penile tissues, and promoted neurite outgrowth in major pelvic ganglia and dorsal root ganglia under high-glucose conditions. Furthermore, bone morphogenetic protein 2 suppressed fibrosis by reducing mouse cavernous endothelial cell fibronectin, collagen 1, and collagen 4 levels under high-glucose conditions. Bone morphogenetic protein 2 modulates neurovascular regeneration and inhibits fibrosis to revive the mouse erection function in diabetic conditions. Our findings propose that the bone morphogenetic protein 2 protein represents a novel and promising approach to treating diabetes-related erectile dysfunction.

Downregulation of the expression of galanin impairs erectile function in hypoandrogenic rats

Abstract Background The relationship between galanin and erectile function under low androgen levels is still unclear. Aim To explore whether a low testosterone level damages the erection of a rat by regulating the expression of galanin and GalR in penile cavernous tissue. Methods Thirty-six male Sprague-Dawley rats, 8 weeks of age, were randomly grouped as follows (n = 6): control, castration, castration + testosterone replacement, control + transfection, castration + transfection, and castration + empty transfection. At 4 weeks after castration, rats in the transfection group were injected with lentivirus carrying the targeting galanin gene (2 × 108 TU/mL, 10 μL) in the corpus cavernosum. After 1 week of injection, the intracavernosal pressure (ICP), mean arterial blood pressure (MAP), nitric oxide (NO), serum testosterone concentration, galanin, GalR1-3, ROCK1, ROCK2, and p-eNOS/eNOS in the rat penile tissues were evaluated. Outcomes ICPmax/MAP and the expression of galanin in the corpus cavernosum in castrated rats were obviously decreased as compared with those in the control rats. Results The castrated rats showed remarkably lower ICPmax/MAP, galanin, GalR1-3, p-eNOS/eNOS, and NO content and markedly higher ROCK1 and ROCK2 in penile tissues than the control group (P &amp;lt; .05). The transfected rats administrated with LV Gal had obviously higher ICPmax/MAP, p-eNOS/eNOS, and NO content and less ROCK1 and ROCK2 protein expression in the corpus cavernosum when compared with the castration group (P &amp;lt; .05). Clinical Translation Upregulating the expression of galanin in the penile corpus cavernosum might be a novel method of treating erectile dysfunction caused by a low androgen level. Strengths and Limitations The conclusions obtained in the animal experiments need to be confirmed in human data. Conclusion The erectile function of hypoandrogen rats might be inhibited by downregulating the level of galanin and GalR1-3, upregulating ROCK1 and ROCK2 levels, and inhibiting the eNOS/NO signaling pathway in penile corpus cavernosum.

CD207 Expression Level is a New Prognostic Marker for Condyloma Acuminatum.

Condyloma cuminata (CA) is a sexually transmitted disease caused by human papillomavirus (HPV) infection, which is prone to recurrence and difficult to cure in the short term. CD207 is a C-type lectin receptor that is specifically expressed on the surface of Langerhans cells (LCs) and is considered as an LC-specific immunohistochemical marker. The main purpose of this study is to explore the correlation between the expression of CD207 in CA skin lesions and the duration of CA disease course and frequency of recurrence, in order to provide new prognostic markers for CA to clinicians. A total of 40 male patients with CA and their skin lesions were collected, as well as 40 healthy male penile tissue samples. The skin lesions of CA were clinically and histologically confirmed by acetic acid test. The expression of CD207 in epidermal tissues was detected using immunohistochemistry. The difference in the number of CD207 positive cells between CA skin lesions and healthy skin controls was compared, and the association between the number of CD207 positive cells in CA skin lesions and the duration of disease course and the frequency of recurrence was determined through Spearman correlation analysis. In CA skin lesions, CD207 positive cells were found to have morphological abnormalities and the number of cells was significantly reduced compared to healthy skin, suggesting that there may be antigen presentation dysfunction in CA skin lesions, which may be the reason for the prolonged and unresolved condition of the disease. The fewer CD207 positive cells in CA skin lesions, the longer the disease course and the more frequent the recurrence, therefore, the expression level of CD207 can be used as a new prognostic marker for predicting the outcome of CA.

(074) Association of Androgen Receptor Signaling in Human Corpus Cavernosum with Serum Testosterone Levels

Abstract Introduction Testosterone (T) plays an important role in properly developing and maintaining male reproductive tissues. The normal serum testosterone levels can vary between 300 to 1000 ng/dL or 10.4 to 34.7 nmol/L. However, it is unclear whether androgen receptor signaling can vary based on serum T levels or the half-life of testosterone therapy. What remains unknown is whether there is a uniform serum T level above which androgen receptor signaling within the tissue remains similar (saturation hypothesis). Objective We first evaluated whether varying serum levels of T alter androgen receptor (AR) signaling in human penile tissue. Then, we evaluated and compared AR signaling in corpus cavernosum tissue of men treated with short-acting vs. long-acting testosterone. We hypothesized that we would find similar AR signaling in the penile tissue of both eugonadal and hypogonadal men as well as those men receiving short and long-acting testosterone. Methods We obtained human corpus cavernosum (hCC) tissue biopsies during penile surgery (for erectile dysfunction) from 14 men. We recorded their serum T levels on week prior to surgery before 10AM. We used western blot to evaluate adrenergic receptor (AR) and VEGF protein levels. Men without erectile dysfunction undergoing penile surgery for Peyronie's disease were used as controls. Results The median age of participants was 61 (IQR 12) and the median T level was 325ng/dL (IQR 201.45). The results of western blot showed expression of similar levels of VEGF in hCC samples from men with T &amp;gt;200ng/dL. However, men with serum T levels lower than 200 ng/dL had decreased expression of VEGF and AR. Conclusions This finding is significant because it demonstrates that serum testosterone beyond a certain threshold may have a minimal impact on erectile function. If a serum testosterone threshold is identified, this threshold could be a valuable target for future studies on the effects of testosterone on ED. It may alter the treatment threshold for men who have ED as their primary symptom of hypogonadism. This data also suggests the saturation value for penile androgen receptors. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Acerus Pharmaceuticals, Boston Scientific, Coloplast, Endo Pharmaceuticals, Empower Pharmacy, Nestle Health, Olympus, Hims, Inc

(227) Analysis of Gene Expression of Vitamin D in Rats in Experimental Model of Peyronie's Disease and Human Penile Tunica Albuginea

Abstract Introduction Peyronie's disease (PD) is a penile connective tissue disorder that results in an inflammatory process, fibrotic plaque in the tunica albuginea (TA) and penile curvature. The molecular changes of fibrotic plaque formation are complex and include different cellular/protein mechanisms. Vitamin D (vitD) is a steroid hormone that has unconventional pathways of action, and among its effects it has already been described that it plays a role in extracellular matrix remodeling, in the immune response and in fibrosis. The literature regarding the action of vitD in the male genital system is scarce, and the gene expression of its receptors has not yet been demonstrated in penile TA. Objective To identify the gene expression of the vitD receptor in penile TA of rats submitted to the experimental model of PD and human penile TA. Methods The experimental model in rats divided into 2 groups (case and control) described before by Cohen et al. Human penile TA was achieved from Peyronie´s surgery cases and cadaveric controls. The antibody anti vitD (VDR D-6: sc-13133, Santa Cruz Biotechnology®, CA, USA) was used. Immunolabeling was carried out using the avidin-biotin-peroxidase complex method, and 3,3′-diaminobenzidine as liquid chromogen. In each case, the immunolabeling was quantified by computer-assisted digital image analysis following the methodology described by Matos et al. PCR real-time analyses followed the MIQE guideline10. One of the fractions of the tissue samples of the TA was stored in RNAholder. The extraction of total RNA from the samples was carried out using the reagent TRIzol®. The reverse transcription was performed using the reverse transcriptase enzyme ImPromII™ to obtain complementary DNA (cDNA). Messenger RNA expression of vitD was obtained by RT-PCR. Quantitative RT-PCR was carried out using pair of oligonucleotides endogenous RPL13a (human samples) and GAPDH (rat samples), known commercially as forward and reverse primers. It was performed statistical analysis with SPSS® version 17.0 (SPSS® Inc; Illinois, USA). Values were expressed as a mean and standard error with a significance level of 95% (p ≤ 0.05). The tests applied were One-Way ANOVA and unpaired t-test with Welch's correction, confirming the similarity between the variances. Sensitivity and specificity analysis was calculated following the Youden index. Ethical approval local committee number 06/2016. Results The vitD mRNA amplifications demonstrated the presence of its receptors in the penile TA in both the control and the case groups, and the expression of the control group was statistically higher (p &amp;lt; 0,0001) when compared to the case group in rats. In human samples, mRNA amplifications demonstrated the presence of its receptors in the penile TA in both the control and the case groups, and the expression of the control group was statistically lower (p &amp;lt; 0,0001) when compared to the case group. Conclusions The presence of vitD receptors (genic and proteic expression) in penile TA of rats Peyronies´ models and human TA was demonstrated and allows future studies regarding its regulation and modulation according to the evolution and phase of PD and vitD serum levels. Disclosure No

(123) Novel Low Intensity Shockwave Therapy Sham Delivery Method (Zero Energy) Assessed Objectively by Grayscale and Doppler Ultrasound Results

Abstract Introduction Most ED LiSWT studies use the IIEF as a primary outcome. While clinically relevant, the IIEF does not directly assess erectile tissue structure. To establish a more objective parameter, we investigated the estimation of hypoechoic area in grayscale ultrasound images of erectile tissue as an objective primary outcome for clinical trials assessing the efficacy of LiSWT. During erection, grayscale imaging of the penis demonstrates corpora cavernosa (CC) that is homogenous in healthy men and heterogeneous to varying degrees in men with ED. A critical aspect of using grayscale ultrasound is to determine that sham treatment reliably results in no significant improvement in erectile tissue structure. Other studies have used sham probes with cotton wadding or non-penetrable gel pads to dampen delivery of shockwave energy flux density, however a small amount of energy may still be delivered. We report a novel sham-controlled methodology in LiSWT RCT for ED. Objective To evaluate erectile tissue heterogeneity as an endpoint in LiSWT sham-controlled trials using grayscale analysis of percent hypoechoic regions in CC after sham delivery of zero energy flux density. Methods A single-blind, sham-controlled, randomized prospective study was performed in men with ED naïve to any kind of acoustic wave therapy. Those meeting inclusion/exclusion completed baseline assessments and were randomized to active or sham within each treatment arm. This interim sub-analysis only examined the sham group (n = 7). For each treatment, the shockwave device was already on in the exam room. When the subject was ready a curtain was suspended above his mid-section blocking the device, and noise cancelling headphones provided. When the probe was placed on the penis, a high-quality recording of shockwave sounds was activated for sham treatment. Twenty weeks after zero energy sham treatment, grayscale and DUS imaging using a 15.4 MHz probe under pharmacologic erection 3-4/4 hardness was performed before unblinding. Grayscale percent hypoechoic regions within the CC were assessed by computer-assisted histometry using Image J. (Fig 1) The IIEF was also administered to all patients at baseline and week 20. Grayscale data were analyzed by two-way repeated measures ANOVA. IIEF-EF scores and bloodflow values (PSV and EDV) were compared using the Wilcoxon matched-pairs signed rank test. Results Compared to baseline, there were no significant changes (p&amp;gt;0.7; see figure) at week 20 in hypoechoic area in the proximal, middle, and distal erect penile shaft. (Fig 2) Furthermore, there were no significant changes in PSV (p&amp;gt;0.15), EDV (p&amp;gt;0.2), or IIEF-EF (p&amp;gt;0.6) from baseline to week 20. Conclusions Using computer-assisted histometry to determine hypoechoic area in grayscale ultrasound images to assess penile erectile tissue heterogeneity, we demonstrated that a novel sham treatment protocol that ensures zero energy delivery does not change erectile tissue integrity. The lack of improvement in PSV, EDV, and IIEF-EF further support our findings with grayscale ultrasound. Future studies will assess the appropriateness of grayscale ultrasound image analysis as a primary endpoint in LiSWT clinical trials. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: TRT, Cynosure.

(231) Endogenous Vasoactive Peptides in the Control of Human Penile Erectile Tissue: Is There a Significance of the Neuropeptide Y (NPY)?

Abstract Introduction Aside from the neurotransmitters of the sympathetic adrenergic system, vasoconstrictor peptides may also play a role in mediating the constant tone of the erectile tissue during penile flaccidity and in returning the erect penis to the flaccid state. Neuropeptide Y (NPY) is present in high concentrations in the male genital tract. The peptide has been shown to induce contraction of isolated human penile tissue (corpus cavernosum, penile arteries) and potentiate its response to noradrenaline. Objective The purpose of our study was to measure in the cavernous and systemic blood of healthy male volunteers the course of NPY through different stages of sexual arousal. Methods Whole blood was drawn from the corpus cavernosum and the cubital vein of 15 healthy male volunteers during penile flaccidity (Fl), tumescence (Tu), rigidity (Ri) and detumescence (Det). Penile tumescence and erection were induced by applying audiovisual and tactile stimulation. Plasma levels of NPY (given in pmol/L) were determined by means of an enzyme-linked immunoassay (ELISA, IBL GmbH, Hamburg, Germany). Results NPY significantly decreased in the cavernous blood with the beginning of sexual arousal, when the flaccid penis became tumescent and finally rigid (Fl: 88.8 ± 35.8, Tu: 62.4 ± 22.7, Ri: 62.3 ± 19.7). In the phase of detumescence, only a slight increase was noted (64.8 ± 23). In the systemic circulation, no pronounced alterations in the concentration of NPY were registered through the different penile stages (Fl: 64.4 ± 27, Tu: 65.8 ± 19, Ri: 59.6-25, Det: 67.6 ± 29.3). Conclusions Our findings are in favor of the hypothesis that NPY could contribute to the maintenance of the resting state of cavernous smooth muscle. It remains to be elucidated whether the peptide may play a role in the pathophysiology of erectile dysfunction. Disclosure No

Expression of Rab25 is down-regulated in the foreskin of children with hypospadias

Hypospadias, a congenital malformation of the penis, is one of the newborns' most common developmental defects. The incidence of hypospadias is increasing yearly, and its pathogenesis is closely related to genetic susceptibility and environmental exposure to endocrine disruptors. Exploring the hypospadias' key molecular regulatory mechanism is crucial to reducing its incidence. To examine the differential expression of Rab25 in hypospadias and normal penile tissue and to identify whether it is a candidate gene for exploring the mechanism of hypospadias. This study included 18 children aged 1-6 years undergoing hypospadias repair surgery at the Children's Hospital of Chongqing Medical University, and foreskin samples were collected. Children diagnosed with cryptorchidism, intersex status, or endocrine abnormalities were excluded from this study. Another 18 children aged 3-8 years with phimosis were included in the control group. The specimens were used for immunohistochemistry, western blotting, immunofluorescence, and polymerase chain reaction to assess the expression of Rab25. Rab25 protein expression was lower in the hypospadias group than in the control group [ (2.101±0.1845), (0.7506±0.1779), p=0.0008<0.05). The hypospadias group showed decreased expression of Rab25 protein in the epithelial cell layer. Rab25 mRNA levels were downregulated in the foreskin of children with hypospadias compared with controls [(1.697±0.2005), (0.7687±0.2130), p=0.0053<0.05)]. Rab25 mRNA and protein expressions in the hypospadias group were significantly downregulated compared with the control group. This was consistent with the results of single-cell sequencing of fetal mice reproductive nodules at 15.5 days of gestation (Zhang Z, Liu Z, Zhang Q, etal., unpublished observations). Our study represents the first report of abnormal Rab25 expression in the foreskin tissue of patients with hypospadias. More detailed research on the relationship between Rab25 and urethral development could be conducted to reveal the molecular mechanism of hypospadias. The expression of Rab25 in foreskin tissue was lower in the hypospadias group than in the control group. Rab25 is involved in the formation of the urethral seam and the occurrence of hypospadias. The potential mechanism by which Rab25 affects the canalization of the urethral plate needs to be further investigated.