Huixi Li

Evaluation of the Effect of Different Doses of Low Energy Shock Wave Therapy on the Erectile Function of Streptozotocin (STZ)-Induced Diabetic Rats

To investigate the therapeutic effect of different doses of low energy shock wave therapy (LESWT) on the erectile dysfunction (ED) in streptozotocin (STZ) induced diabetic rats. SD rats (n = 75) were randomly divided into 5 groups (normal control, diabetic control, 3 different dose LESWT treated diabetic groups). Diabetic rats were induced by intra-peritoneal injection of STZ (60 mg/kg) and rats with fasting blood glucose ≥ 300 mg/dL were selected as diabetic models. Twelve weeks later, different doses of LESWT (100, 200 and 300 shocks each time) treatment on penises were used to treat ED (7.33 MPa, 2 shocks/s) three times a week for two weeks. The erectile function was evaluated by intracavernous pressure (ICP) after 1 week washout period. Then the penises were harvested for histological study. The results showed LESWT could significantly improve the erectile function of diabetic rats, increase smooth muscle and endothelial contents, up-regulate the expression of α-SMA, vWF, nNOS and VEGF, and down- regulate the expression of RAGE in corpus cavernosum. The therapeutic effect might relate to treatment dose positively, and the maximal therapeutic effect was noted in the LESWT300 group. Consequently, 300 shocks each time might be the ideal LESWT dose for diabetic ED treatment.

Novel Therapeutic Approach for Neurogenic Erectile Dysfunction: Effect of Neurotrophic Tyrosine Kinase Receptor Type 1 Monoclonal Antibody

BACKGROUND Erectile dysfunction (ED) is a major health issue in aged populations, and neurogenic ED is particularly difficult to treat. Novel therapeutic approaches are needed for treatment of neurogenic ED of peripheral origin. OBJECTIVE To investigate the therapeutic effects of a neurotrophic tyrosine kinase receptor type 1 monoclonal antibody (TrkA-mAb) on erectile function and sexual behavior in a rat model of cavernous nerve injury (CNI). DESIGN, SETTING, AND PARTICIPANTS In one experiment, 84 male rats were randomly assigned to seven groups. The groups underwent either CNI or sham surgery, subsequent injection into the major pelvic ganglion (IMPG) of phosphate-buffered saline (PBS), an immunoglobulin G (IgG) control, or TrkA-mAb, and then intracavernosal (IC) injection of either PBS or varying TrkA-mAb concentrations immediately after surgery and then 1 wk later. Erectile function was assessed and histologic/molecular analyses were performed at 6 wk after surgery. In a second experiment, 36 male rats were randomly divided into three groups. The groups underwent CNI or sham surgery and then IC injection of PBS, IgG, or TrkA-mAb immediately after surgery and for 5 wk thereafter. At 6 wk after surgery, the performance of the rats in sexual behavior tests was videotaped. INTERVENTION CNI or sham surgery; IMPG of PBS, IgG, or TrkA-mAb; IC injection of PBS or TrkA-mAb. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS The intracavernous pressure response to cavernous nerve electrostimulation was measured and midpenile cross-sections were histologically examined. Western blotting (WB) of cavernous tissue protein was performed. Rats were assessed for chasing, mounting, intromission, and ejaculation behaviors during sexual behavior tests. The data were analyzed using one-way analysis of variance followed by the Tukey-Kramer t test. RESULTS AND LIMITATIONS Recovery of erectile function of varying degrees was observed in the TrkA-mAb groups. TrkA-mAb treatment significantly suppressed tyrosine hydroxylase-positive nerve fibers in the corpus cavernosum and enhanced neuronal nitric oxide synthase-positive fibers in the dorsal nerve. The ratio of smooth muscle to collagen in the corpus cavernosum was significantly improved in TrkA-mAb treatment groups compared to PBS vehicle and IgG control groups. WB confirmed these biological changes. There was a nonsignificant increase in the average number of intromissions and ejaculations in the TrkA-mAb group. The study limitations include small sample size, variability in sexual behavior, lack of data on the neuromuscular mechanism involved, and lack of information of the role of neurotrophins or cytokines in regeneration. CONCLUSIONS TrkA-mAb successfully inhibits sympathetic nerve regeneration, leads to parasympathetic nerve regeneration, and has therapeutic effects on ED and sexual behavior disorder in a rat model of CNI. PATIENT SUMMARY This report provides strong evidence that a neurotrophic tyrosine kinase receptor type 1 monoclonal antibody (TrkA-mAb) inhibits sympathetic nerve regeneration, leads to parasympathetic nerve regeneration, and has therapeutic effects on erectile dysfunction and sexual behavior disorder in a rat model of cavernous nerve injury. The results raise the possibility that human patients with neurogenic erectile dysfunction may respond to TrkA-mAb in a manner that parallels the response seen in our rodent study.

Urethral musculature and innervation in the female rat.

The urethral sphincter and urethral muscle innervation are critically involved in maintaining continence, especially in the female. However, the urethral muscle type and distribution, as well as the urethral nerves are far from being well documented. Our aim was to clearly identify the distribution of urethral striated muscle, smooth muscle, and urethral nerves.

Prophylactic protective effects and its potential mechanisms of icarisideii on streptozotocin induced spermatogenic dysfunction

The aim of this study was to investigate the effects of IcarisideII(ICAII) on the prevention of streptozotocin (STZ) induced spermatogenic dysfunction. Forty male Sprague-Dawley rats received intraperitoneal injection of STZ (55 mg/kg) and were equally randomized to gavage feeding of vehicle (the vehicle group) or ICAII (0.5, 1.5 or 4.5 mg/kg/day, respectively). Ten normal rats received vehicle and served as control. Four weeks later, sperm parameters, histopathological changes, testicular lipid peroxidation, antioxidant enzyme activities, and apoptosis index (AI) were evaluated. Results showed that ICAII treatment resulted in a significant recovery of sperm parameters and histopathological changes relative to the vehicle group (p < 0.05). In the vehicle group, antioxidant enzyme activities and the expression of Sertoli cell Vimentin filaments obviously decreased, while lipid peroxidation and AI significantly increased as compared with the control group (p < 0.05). Following ICAII treatment, corrective effects on these items towards normal levels were observed. The results suggested that ICAII has beneficial effect on the preservation of spermatogenic function in the STZ-induced diabetic rats. The mechanisms might be related to its improvement of antioxidant enzyme activities, preservation of the protein expression and apical extensions of Vimentin filaments, and anti-apoptosis capability.

Transgenic animal model for studying the mechanism of obesity-associated stress urinary incontinence

To study and compare the function and structure of the urethral sphincter in female Zucker lean (ZL) and Zucker fatty (ZF) rats and to assess the viability of ZF fats as a model for female obesity‐associated stress urinary incontinence (SUI).

Comparison of Topical Hemostatic Agents in a Swine Model of Extremity Arterial Hemorrhage: BloodSTOP iX Battle Matrix vs. QuikClot Combat Gauze

BloodSTOP iX Battle Matrix (BM) and QuikClot Combat Gauze (CG) have both been used to treat traumatic bleeding. The purpose of this study was to examine the efficacy and initial safety of both products in a swine extremity arterial hemorrhage model, which mimics combat injury. Swine (37.13 ± 0.56 kg, NBM = 11, NCG = 9) were anesthetized and splenectomized. We then isolated the femoral arteries and performed a 6 mm arteriotomy. After 45 s of free bleeding, either BM or CG was applied. Fluid resuscitation was provided to maintain a mean arterial pressure of 65 mmHg. Animals were observed for three hours or until death. Fluoroscopic angiography and wound stability challenge tests were performed on survivors. Tissue samples were collected for histologic examination. Stable hemostasis was achieved in 11/11 BM and 5/9 CG subjects, with recovery of mean arterial pressure and animal survival for three hours (p < 0.05, Odds Ratio (OR) = 18.82 (0.85–415.3)). Time to stable hemostasis was shorter for the BM-treated group (4.8 ± 2.5 min vs. 58 ± 20.1 min; Median = 2, Interquartile Range (IQR) = 0 min vs. Median = 60, IQR = 120 min; p < 0.05) and experienced longer total stable hemostasis (175.2 ± 2.5 min vs. 92.4 ± 29.9 min; Median = 178, IQR = 0 min vs. Median = 120, IQR = 178 min; p < 0.05). Post-treatment blood loss was lower with BM (9.5 ± 2.4 mL/kg, Median = 10.52, IQR = 13.63 mL/kg) compared to CG (29.9 ± 9.9 mL/kg, Median = 29.38, IQR = 62.44 mL/kg) (p = 0.2875). Standard BM products weighed less compared to CG (6.9 ± 0.03 g vs. 20.2 ± 0.4 g) (p < 0.05) and absorbed less blood (3.4 ± 0.8 g vs. 41.9 ± 12.3 g) (p < 0.05). Fluoroscopic angiography showed recanalization in 5/11 (BM) and 0/5 (CG) surviving animals (p = 0.07, OR = 9.3 (0.41–208.8)). The wound stability challenge test resulted in wound re-bleeding in 1/11 (BM) and 5/5 (CG) surviving animals (p < 0.05, OR = 0.013 (0.00045–0.375)). Histologic evidence indicated no wound site, distal limb or major organ damage in either group. BM is more effective and portable in treating arterial hemorrhage compared to CG. There was no histologic evidence of further damage in either group.

AB101. Therapeutic effect of low intensity pulsed ultrasound in stress urinary incontinence

Objective Stress urinary incontinence, a major type of urinary incontinence, increases with age and is often developed after partum injury. Low intensity pulsed ultrasound (LIPUS) has been investigated in the treatment of many diseases showing its ability of restoring soft tissue injury. We investigated the therapeutic effect of low intensity pulsed ultrasound in stress urinary incontinence. Methods Thirty-two Sprague Dawley rats in SUI group underwent vaginal distension (VD) and bilateral ovariectomy mimicking partum injury. Eight rats served as mock operation control. Eight rats each in SUI group was treated with low-dosage LESW (0.03 mJ/mm2), medium-dosage LESW (0.06 mJ/mm2), or high-dosage LESW (0.09 mJ/mm2). The rest eight rats served as none-treatment group. For functional study, leak point pressure test (LPP) was performed 2 weeks after the last LESW. Masson trichrome staining was performed to validate the pathological changes. Results The LPP was restored in medium-dosage LESW and high-dosage LESW groups, but not in low-dosage LESW group. More robust striated muscle regeneration was found in these two groups comparing with the none-treatment group. Conclusions LIPUS ameliorate the symptom of SUI via activating striated muscle regeneration.

Low Intensity Pulsed Ultrasound Influences the Myogenic Differentiation of Muscle Satellite Cells in a Stress Urinary Incontinence Rat Model

OBJECTIVE To investigate the therapeutic effect of low intensity pulsed ultrasound (LIPUS) in a stress urinary incontinence (SUI) rat model and its influence on myogenic satellite cells. METHODS Fifty Sprague-Dawley rats underwent vaginal distension and bilateral ovariectomy mimicking partum injury and menopause to construct SUI models, which were further randomized into 100 mW/cm2 LIPUS, 200 mW/cm2 LIPUS, 300 mW/cm2 LIPUS, and none-treatment control subgroups with 10 rats per subgroup. Ten rats served as mock operation control. Leak point pressure and bladder capacity were recorded 1 week after LIPUS treatment. Immunofluorescence staining and Western blot were performed to examine histological changes, myodifferentiation, and signaling pathway. RESULTS Here,we found the leak point pressure and bladder capacity were restored in 200 mW/cm2 LIPUS and 300 mW/cm2 LIPUS groups, but not in 100 mW/cm2 LIPUS group. More robust striated muscle regeneration was observed in 200 mW/cm2 LIPUS group comparing with the SUI none-treatment group. Moreover, we found LIPUS activated the myodifferentiation of muscle satellite cells, which is correlated to p38 phosphorylation level. CONCLUSION LIPUS restored the leak point pressure and bladder capacity, and activated satellite cell myodifferentiation in SUI rat model.

AB238. Effect of Icariside II on miR-126 pathway on human cavernous endothelial cells exposed to a diabetic-like environment

Background To investigate the status of miR-126 and its targeting Spred1 under the stimulation of glucose and Age-BSA and to explore the effect of icariside II (ICA II) on the diabetic endothelial dysfunction of human cavernous endothelial cell (HCECs) by using the miR-126 pathway. Methods Purified HCECs were divided into three groups: normal group + BSA (NC group), Glucose + Age − BSA group (DM group), ICA II treatment group (DM + ICA II group). Western blot to detect the expression of eNOS, RAGE protein expression so as to make sure the success of model construction; immunofluorescence assay to study the proliferation of (HCECs); real time PCR to detect the expression of miR-126 and Spred1; western blot to detect the expression of the Spred1/c-Raf/MEK1/2/Erk1/2. Tube Formation Assay and Scratch assay were performed to detect the angiogenesis of HCECs under the diabetic-like environment. Results Under the model, the expression of eNOS in DM group significantly reduced compared with that of NC group and the expression of RAGE in DM group is significantly increased compared with that of NC group (P<0.05), but the DM + ICA II group showed higher eNOS expression and lower RAGE expression compared with those in the DM group. The Ki67 expression in DM group is lowered than that in NC group; whereas the Ki67 expression in DM + ICA II group is higher when compared with that in DM group. The expression of miR-126 in DM group is significantly reduced compared with that of NC group but the DM + ICA II group showed higher miR-126 expression compared with that in the DM group. Western blot results showed Spred1 expression increased under the diabetic condition and its downstream target genes c-Raf/MEK1/2/Erk1/2 expression decreased obviously, but ICA II adding into the DM group could reverse these results effectively. Tube Formation Assay and Scratch assay also showed ICA II could promote the tube formation and cell proliferation in impairment of endothelial dysfunction of DM group. Conclusions Under the simulation of Age-BSA and glucose, HCECs occurred the endothelial dysfunction and the angiogenesis were repressed; ICA II could restore the HCECs functions by miR-126/Spred1/c-Raf/MEK1/2/Erk1/2 pathway. ICA II may be a promising therapeutic compound to treat endothelial dysfunction in the future.

AB239. Icariside II enhances endothelial nitric oxide synthase expression by suppressing miR-155 in diabetic-like human cavernous endothelial cells

Background To investigate the role of Icariside II (ICA II) on endothelial nitric oxide synthase (eNOS) expression regulated by miR-155, the human cavernous endothelial cells (HCECs) were exposed to a diabetic-like environment and treated with ICA II. Methods HCECs were treated with 200 µg/mL BSA as the Normal Control group (NC), with 200 µg/mL AGE-BSA plus 250 mg/dL glucose as the diabetes mellitus (DM) group, or with an addition of ICA II in DM group as the treatment group (DM+ICA II). Bioinformatics were first used to predict miRNAs targeting eNOS gene and then potential candidates including miR-155, miR-543, miR-31, miR-429, miR-200b were further verified by real-time PCR in a diabetic-like condition. Expressions levels of miRNAs, eNOS and the receptor for advanced glycation end products (RAGE) were performed by Real-time PCR; Protein expression levels of eNOS and RAGE were analyzed by western blot; nitric oxide (NO) content was detected by DAF-FM DA probe and NaNO2 standard curve methods. Results The expression of miR-155 in DM group is significantly higher than that that in the normal control (NC) group, whereas this phenomenon was effectively reversed by ICA II treatment. Furthermore, the miR-155 targeting gene eNOS and its consequent NO product were significantly reduced in DM group, while these changes were also recovered after ICA II treatment. Conclusions In this study, we demonstrated that ICA II could promotes eNOS mRNA and protein levels by suppressing miR-155 in HCECs exposed to a diabetic-like environment.