[Effects of different low-dose of insulin glargine on antioxidation of organs in burned rats with delayed resuscitation].

OBJECTIVE: To study the antioxidant protective effects of different low-dose of insulin glargine on organs of burned rats with delayed resuscitation. METHODS: Forty male Sprague-Dawley (SD) rats were randomly divided into sham group, delayed resuscitation control group, and insulin glargine 0.5, 1.0, and 2.0 U groups, with 8 rats in each group. The rats were immersed in hot water (95.0±0.5) centigrade for 15 s to establish the third-degree scald model with 30% total body surface area. The rats in the sham group were immersed in a 37 centigrade water bath for 15 s. Insulin glargine (0.5, 1.0, 2.0 U×kg-1×d-1) was injected subcutaneously in corresponding insulin glargine group 2 hours after injury, and the same amount of normal saline was injected intraperitoneally in the delayed resuscitation control group. Intraperitoneal injection of normal saline 40 mL/kg simulated delayed resuscitation 6 hours after injury in all groups. Abdominal aortic blood samples, heart and kidney tissue were collected immediately after simulating burn in the sham group, and 24 hours after burn in other four groups. The blood glucose, myocardial enzymes [lactate dehydrogenase (LDH), creatine kinase (CK), α-hydroxybutyrate dehydrogenase (α-HBDH), and aspartate aminotransferase (AST)] and renal function indexes [blood urea nitrogen (BUN) and serum creatinine (SCr)] were measured by spectrophotometry, and the isoenzyme MB of creatine kinase (CK-MB) level was determined by immunosuppression method to evaluate the effects of different low-dose insulin glargine intervention on blood glucose, cardiac and renal functions in scalded rats with delayed resuscitation. The oxidative and antioxidant indices [xanthine oxidase (XOD), myeloperoxidase (MPO), copper-zinc superoxide dismutase (CuZn-SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC)] from the heart and kidney tissues of rats were detected by spectrophotometry to analyze the antioxidant effects of different low-dose insulin glargine interventions. RESULTS: Compared with the sham group, the blood glucose of the rats in the delayed resuscitation control group was significantly increased, the heart and kidney functions were significantly reduced, the oxidation capacity was enhanced, and the antioxidant indicators were significantly reduced. After the intervention of insulin glargine, with the increase of insulin glargine dose, the blood glucose, myocardial enzyme and renal function indicators of rats showed a gradual downward trend, the oxidation indicators continued to decrease, and the antioxidant indicators showed a gradual upward trend. When the dose was 2.0 U×kg-1×d-1, the blood glucose, LDH, CK, CK-MB, α-HBDH, AST, BUN, SCr, XOD and MPO were significantly lower than those in the delayed resuscitation control group [blood glucose (mmol/L): 5.91±0.25 vs. 11.76±0.36, LDH (U/L): 3 332.12±51.61 vs. 5 008.94±490.12, CK (kU/L): 0.49±0.03 vs. 0.85±0.04, CK-MB (U/L): 125.40±12.19 vs. 267.52±11.63, α-HBDH (U/L): 122.99±5.37 vs. 240.85±13.99, AST (U/L): 11.95±1.81 vs. 17.87±1.57, BUN (mmol/L): 4.72±0.15 vs. 7.16±0.34, SCr (µmol/L): 87.11±6.51 vs. 137.50±11.36, XOD (U/g): 166.29±3.27 vs. 204.90±4.82 in heart tissue, 63.51±1.46 vs. 79.69±1.75 in kidney tissue, MPO (U/g): 1.05±0.02 vs. 1.55±0.06 in heart tissue, 1.04±0.04 vs. 1.87±0.01 in kidney tissue, all P < 0.05], and CuZn-SOD, CAT, GSH-Px and T-AOC were significantly higher than those in the delayed resuscitation control group [CuZn-SOD (kU/g): 82.95±2.69 vs. 56.52±2.26 in heart tissue, 94.50±2.73 vs. 62.02±1.66 in kidney tissue, CAT (U/g): 36.07±2.01 vs. 15.15±2.22 in heart tissue, 184.49±4.53 vs. 156.02±3.96 in kidney tissue, GSH-Px (kU/g): 231.93±8.03 vs. 179.48±3.15 in heart tissue, 239.63±7.30 vs. 172.20±2.09 in kidney tissue, T-AOC (kU/g): 4.85±0.23 vs. 2.71±0.11 in heart tissue, 5.51±0.08 vs. 3.50±0.07 in kidney tissue, all P < 0.05]. CONCLUSIONS: Different low-dose of insulin glargine (≤ 2.0 U×kg-1×d-1) could exert antioxidant protection on the heart and kidney of rats with delayed resuscitation after burns, with a dose-dependent manner.

Fabrication of Multiple Parallel Microchannels in a Single Microgroove via the Heating Assisted MIMIC Technique.

For the first time, multiple parallel microchannels in a single microgroove have been fabricated by the heating-assisted micromolding in capillaries technique (HAMIMIC). Microchannel development, cross-sectional shape, and length were all explored in depth. The factors affecting the cross-sectional shape and length of the double-microchannel were also discussed. Finally, a special-shaped PDMS guiding mold was designed to control the cross-sectional shape and length of multiple parallel microchannels for controlled growth. The HAMIMIC technique provides a low-cost, straightforward, and repeatable way to create multiple parallel microchannels in a single microgroove, and will promote the progress of bifurcated vessels and thrombus vessels preparation technology.

[Effects of medium and long-term insulin pretreatment on the activity of main oxidase and antioxidant enzyme in the myocardium of burned rats with delayed fluid resuscitation].

OBJECTIVE: To observe the effects of medium and long-term insulin pretreatment on the activity of main oxidase and antioxidant enzyme in the myocardium of burned rats with delayed fluid resuscitation. METHODS: According to random number table method, forty male Sprague-Dawley (SD) rats were divided into pseudo-burn group, burn delayed resuscitation group, insulin glargine pretreatment group and neutral protamine hagedorn (NPH) insulin pretreatment group, with 10 rats in each group. 30% total body surface area (TBSA) as III degree scald model was prepared by bathing the back of rats in 95 centigrade hot water for 15 s; the rats in the pseudo-burn group were immersed in 37 centigrade warm water for 15 s as control. Insulin glargine pretreatment group, NPH insulin pretreatment group and burn delayed resuscitation group were injected subcutaneously with insulin glargine, NPH insulin, and normal saline 1.0 U×kg-1×d-1 2 hours after injury, and intraperitoneal injection of normal saline 40 mL/kg simulated delay resuscitation 6 hours after injury. The pseudo-burn group didn't receive medicine and delayed resuscitation. Abdominal aortic blood samples and heart tissue were collected immediately after simulating scald in the pseudo-burn group, and 24 hours after scald in three burn groups. Blood glucose, xanthine oxidase (XOD), myeloperoxidase (MPO), CuZn-superoxide dismutase (CuZn-SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) of the heart tissue were determined by spectrophotometry. RESULTS: Compared with the pseudo-burn group, the burn delayed resuscitation group have significantly higher blood glucose level and the XOD and MPO in the heart tissue, while significantly lower CuZn-SOD, CAT, and GSH-Px in the heart tissue. Compared with the burn delayed resuscitation group, insulin glargine pretreatment group and NPH insulin pretreatment group have lower blood glucose level and heart tissue XOD [blood glucose (mmol/L): 6.37±1.22, 6.66±1.45 vs. 9.47±0.80; XOD (U/g): 271.93 (261.59, 275.91), 285.32 (251.96, 297.29) vs. 363.37 (354.12, 377.76), all P < 0.05], while significantly higher heart tissue CuZn-SOD, CAT, and GSH-Px [CuZn-SOD (U/g): 0.13±0.01, 0.14±0.01 vs. 0.10±0.01; CAT (U/g): 29.17±7.28, 27.16±7.37 vs. 18.36±4.53; GSH-Px (U/g): 0.33 (0.16,0.41), 0.30 (0.17,0.41) vs. 0.07 (0.04,0.11), all P < 0.05]. MPO activity in insulin glargine pretreatment group was significantly lower than that in burn delayed resuscitation group (U/g: 0.016±0.002 vs. 0.020±0.002, P < 0.05), but there was no significant difference between insulin pretreatment group and NPH insulin pretreatment group (U/g: 0.019±0.003 vs. 0.020±0.002, P > 0.05). There was no significant difference in the blood glucose, and activities of XOD, MPO, CAT, GSH-Px between insulin glargine pretreatment group and NPH insulin pretreatment group, but the activity of CuZn-SOD in NPH insulin pretreatment group was further higher than that in insulin glargine pretreatment group (U/g: 0.14±0.01 vs. 0.13±0.01, P < 0.05). CONCLUSIONS: Medium and long-term insulin pretreatment can improve the antioxidant capacity of myocardium in delayed resuscitation rats after burns, inhibit the production of reactive oxygen species and improve the scavenging capacity of reactive oxygen species. However, only CuZn-SOD activity is different between the two groups, and further study needs to be carried out to determine whether it is related to the type if insulin.

[Effects of low-dose insulin on oxidation-reduction in rat organs after major burns].

OBJECTIVE: To explore the effects of low-dose insulin on oxidation-reduction of heart and kidney in rats immediately after severe burns. METHODS: Twenty four male Sprague-Dawley (SD) rats were randomly divided into 3 groups of sham burn, burn and treatment (n = 8 each). The sham burn group was placed into 37 °C warm water for 15 seconds to simulate burn process and received no fluid replacement. The burn and treatment groups were immersed into (95 ± 0.5) °C hot water for 15 seconds to make a rat model of 30% total burn surface area, III degree burn injury and immediately received an intraperitoneal injection of physiological saline (40 ml/kg). At the same time, a subcutaneous injection of insulin (1.0 U×kg(-1)×d(-1)) was given in the treatment group and a subcutaneous injection of same-volume physiological saline in the burn group. The rats were sacrificed after 24 post-scald hours (PSH). Abdominal aortic blood was collected for an analysis of blood glucose. The oxidation and antioxidation parameters of heart and kidney, such as malondialdehyde (MDA), xanthine oxidase (XO), myeloperoxidase (MPO), total superoxide dismutase (T-SOD), superoxide dismutase 1, 2 (SOD1, 2), catalase (CAT) and glutathione peroxidase (GPx), were detected by spectrophotometry. RESULTS: Compared with the sham burn group, MDA content, XO and MPO activities of heart and kidney were significantly higher in the burn group (all P < 0.05); in the treatment group, MDA content and XO activity of heart were significantly lower than the burn group ((0.85 ± 0.07) vs (1.11 ± 0.07) nmol/mg, (69.72 ± 1.94) vs (77.21 ± 2.10) U/g) while the MPO activities of heart and kidney were significantly lower (all P < 0.05).compared with the sham burn group, the activities of T-SOD, CAT, GPx of heart and kidney were significantly lower in the burn group, SOD1 activity of kidney was significantly lower, but SOD2 activity of kidney was significantly higher while SOD2 activity of heart was significantly lower (all P < 0.05); Compared with the burn group, the activities of T-SOD and SOD1 of heart in the treatment group were significantly higher ((83.5 ± 2.5) vs (79.6 ± 3.2), (62.8 ± 2.3) vs (58.8 ± 3.0) U/mg), CAT and GPx activity of heart and kidney were significantly higher (all P < 0.05). Compared with the sham burn group, blood glucose in the burn and treatment group were significantly higher ((7.81 ± 0.30), (7.19 ± 0.22) vs (6.30 ± 0.24) mmol/L) and blood glucose in the treatment group was significantly lower than the burn group((7.81 ± 0.30) mmol/L)(all P < 0.05). CONCLUSIONS: During an early stage, a low-dose insulin may intervene in heart tissue lipid peroxidation of severely burned rats. And differences exist in the effects of oxidation-reduction between heart and kidney.

Activation of VEGF and ERK1/2 and improvement of urethral function by adipose-derived stem cells in a rat stress urinary incontinence model.

OBJECTIVE: To investigate the injected autologous adipose-derived stem cells (ADSCs) in improving stress urinary incontinence in a rodent model of parturition-related stress incontinence and the possible mechanism. METHODS: The 40 rats were developed stress urinary incontinence models by postpartum balloon dilation of the vagina for 4 hours followed by bilateral ovariectomy. ADSCs were isolated from the peri-ovarian fat and labeled with thymidine analog 5-ethynyl-2-deoxyuridine (EdU). Twenty stress urinary incontinence rats received peri-urethral injection of phosphate-buffered saline as the negative controls and the other 20 stress urinary incontinence rats received peri-urethral injection of EdU-labeled ADSCc. Twenty control rats underwent sham ovariectomy without balloon dilation and served as positive controls. Four weeks later, voiding function was assessed by cystometry. Urethral histologic examination (Masson trichrome stain, picrosirius red stain, Hart elastin stain, Gordon and Sweet stain, and immunohistochemical stain) and Western blot were performed on urethral tissues. RESULTS: Both leak point pressure and bladder capacity were significantly increased in ADSC-treated rats, compared to the balloon-injured ovariectomized rats. Histologic examination revealed normalized appearance of the fibromuscular structure of the urethra as well as increased peri-urethral blood vessel density in ADSC-treated rats. On Western blot, vascular endothelial growth factor and P-extracellular signal-regulated kinases (ERKs)1/2 protein was expressed at a higher rate in tissues from ADSC-treated rats compared to phosphate-buffered saline-treated rats. CONCLUSION: Peri-urethral injection of ADSCs is associated with more normal urinary function and urethral structure in rats with parturition-related incontinence. The activation of vascular endothelial growth factor and ERK1/2 may be responsible for the paracrine effects from ADSCs.

Effects of icariside II on improving erectile function in rats with streptozotocin-induced diabetes.

Icariin and icariside II (ICA II), 2 active components isolated from herba epimedii, have a closely structural relationship. There is evidence that icariin may be useful in the treatment of erectile dysfunction (ED); however, the study on the therapeutic efficacy of ICA II on ED is currently scant. We investigated the effects of ICA II on improving erectile function of rats with streptozocin-induced diabetes. Fifty 8-week-old Sprague-Dawley rats were randomly distributed into normal control and diabetic groups. Diabetes was induced by a one-time intraperitoneal injection of streptozocin (60 mg/kg). Three days later, the diabetic rats were randomly divided into 4 groups including a saline-treated placebo arm and 3 ICA II-treated models (1, 5, and 10 mg/kg/d). After 3 months, penile hemodynamics was measured by cavernous nerve electrostimulation (CNE) with real time intracorporal pressure assessment. Penises were harvested with subsequent histological examination (picrosirius red stain, Hart elastin stain, and immunohistochemical stain) and Western blots to explore the expression of the nitric oxide-cyclic guanosine monophosphate (NO-cGMP) and transforming growth factor ß1 (TGFß1)/Smad2 signaling pathways. Diabetes significantly attenuated erectile responses to CNE. Diabetic rats had decreased corpus cavernosum smooth muscle/collagen ratio and endothelial cell content relative to the control group. The ratio of collagen I to III was significantly lower in the corpora of diabetic rats; furthermore, cavernous elastic fibers were fragmented in the diabetic animals. Neuronal nitric oxide synthase (nNOS), endothelial nitric oxide synthase, and vascular endothelial growth factor were expressed at lower levels in the diabetic group; ICA II-treated diabetic rats had higher expression in the penis relative to placebo-treated diabetic animals. Both the TGFß1/Smad2/connective tissue growth factor (CTGF) signaling pathway and apoptosis were down-regulated in the penis from ICA II-treated rats. ICA II treatment attenuates diabetes-related impairment of penile hemodynamics, likely by increasing smooth muscle, endothelial function, and nNOS expression. ICA II could alter corpus cavernosum fibrous-muscular pathological structure in diabetic rats, which could be regulated by the TGFß1/Smad2/CTGF and NO-cGMP signaling pathways.

Pathology of urethral fibromuscular system related to parturition-induced stress urinary incontinence and TGF-ß1/Smad pathway.

To study pathological changes of fibromuscular system and the role of TGF-ß1/Smad pathway in the urethra of a parturition-induced stress urinary incontinence (SUI) rat model. Twenty-eight 8-week-old Sprague-Dawley female rats at gestational day 16 were used and randomized into two groups: sham group and SUI group. After delivery, rats in the SUI group underwent postpartum vaginal balloon dilation and bilateral ovariectomy. 1 month after ovariectomy, urodynamics was assessed. Histological examination (Masson's trichrome stain, picrosirius red stain, Hart's elastin stain, Gordon & Sweet's stain, and immunohistochemical stain) and Western blot were performed on urethral tissues. Both leak point pressure and maximal bladder capacity were significantly decreased in the balloon-injured ovariectomized rats, compared with the sham rats. Muscle was significantly decreased in the urethra of SUI rats compare with sham rats. Collagen I/III and reticular fibers from SUI group were also significantly lower than sham group. Meanwhile, elastic fibers and reticular fibers showed fragmentation and disorganization indicating impairment in the fibromuscular system in SUI rats. TGF-ß1, MMP-9, and phosphorylated Smad2 (p-Smad2) were expressed significantly higher in SUI than in sham rats. Simulated birth trauma and menopause induced an upregulation of the TGF-ß1/Smad pathway and impairment of the fibromuscular system in the urethra.

The TGF-ß1/Smad/CTGF pathway and corpus cavernosum fibrous-muscular alterations in rats with streptozotocin-induced diabetes.

Diabetes-associated erectile dysfunction is associated with increased extracellular matrix deposition and reduced smooth muscle content in the corpus cavernosum. The mechanisms of these processes are not well understood. In this study, we investigated fibromuscular changes in the corpus cavernosum of rats with streptozotocin-induced diabetes to determine the mechanisms underlying pathologic changes in penile structure and function. Forty 8-week-old Sprague-Dawley rats were randomly distributed into control and diabetic groups. Diabetes was induced by a one-time intraperitoneal injection of streptozotocin 60 mg/kg. Twelve weeks later, erectile function was measured by cavernous nerve electrostimulation with real-time intracorporal pressure assessment. The penis was harvested for histologic examination (Masson trichrome stain, picrosirius red stain, Hart elastin stain, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling, and immunohistochemistry) and Western blot. Diabetes significantly attenuated erectile response to cavernous nerve electrostimulation. Diabetic animals exhibited a decreased smooth muscle/collagen ratio in the corpus cavernosum. The ratio of collagen I to II fibers was significantly lower in the corpora of diabetic rats compared with controls. Cavernous elastic fibers were fragmented in diabetic rats. There was up-regulation of the transforming growth factor ß1/Smad/connective tissue growth factor signaling pathway in diabetic rats. Phospho-Smad2 expression was higher in smooth muscle cells and fibroblasts of diabetic rats, as was the apoptotic index. The up-regulation of the transforming growth factor ß1/Smad/connective tissue growth factor signaling pathway might play an important role in diabetes-induced fibrous-muscular structural changes and deterioration of erectile function.

Depletion of tissue factor suppresses hepatic metastasis and tumor growth in colorectal cancer via the downregulation of MMPs and the induction of autophagy and apoptosis.

Tissue factor (TF) is a significant risk factor for hepatic metastasis in patients with colorectal cancer (CRC). However, the mechanism by which TF promotes hepatic metastasis in CRC remains elusive. In this study, we first confirmed that TF expression was significantly correlated with lymph node metastasis, hepatic metastasis and TNM staging in clinical CRC samples, and found that TF expression in colon cancer cell lines was correlated with the invasion ability. Next, by employing TF-overexpressing LOVO cell line as a model we demonstrated that lentivirus mediated knockdown of TF suppressed the migration and invasion of LOVO cells in vitro, and hepatic metastasis of colorectal cancer in nude mice orthotopic model. Mechanistically, we found that TF knockdown decreases colony formation ability and induced autophagy and apoptosis of LOVO cells, and this was at least partly mediated by the activation of unfolded protein response/PERK signaling. In conclusion, our data provide new insight into hepatic metastasis of CRC. Agents targeting TF should be developed as adjuvant therapeutics for CRC metastasis.

[Diagnosis and treatment of Kawasaki disease in burn children].

We try to discuss the relationship between burn and Kawasaki disease (KD), and to study the diagnosis and treatment of KD in burn children. The medical records of one burn child with KD from our pediatric ward together with those of 5 burn children with KD retrieved from foreign literature were analyzed. The clinical features of KD, including bilateral nonexudative conjunctivitis, erythema of the lips and oral mucosa, skin rash, cervical lymphadenopathy, changes in the distal part of extremities, were enrolled in the study. Six patients were male and younger than 5 years old, with 2 suffering from mild burn, 2 with moderate burn, and 2 with severe burn. Two days after second degree burn, all burn children had fever and skin rash with 4 or 5 clinical symptoms and signs of KD. Among them, coronary artery dilatation was found in 1 case as detected by echocardiography, positive wound culture was found in 2 cases, negative blood culture was found in 6 cases. All patients were given high-dose gamma globulin or (and) aspirin within 10 days after the first fever, followed by control and amelioration of the disease. We conclude that the pathogenesis of KD may be related with burn wound and reabsorption of edema. KD may be suspected in burn children younger than 5 years when they had fever and skin rash at the same time.

Effects of icariin on improving erectile function in streptozotocin-induced diabetic rats.

INTRODUCTION: Icariin has been shown to improve penile hemodynamics in animal models of erectile dysfunction from cavernous nerve injury and castration. The effects of icariin on penile hemodynamics in diabetic animals remain to be determined. Transforming growth factor ß1 (TGFß1) has been implicated in the pathogenesis of diabetes-related erectile dysfunction. AIM: The aim of this study was to investigate the effects of icariin in the penis of streptozotocin (STZ)-induced diabetic rat. METHODS: Two-month-old Sprague-Dawley male rats received one-time intraperitoneal (IP) STZ (60 mg/kg) or vehicle injection after a 16-hour fast. Three days later, the STZ-induced diabetic rats were randomly divided into four groups and were treated with daily gavage feedings of a 50:50 mix of normal saline and dimethyl sulfoxide (DMSO) or icariin dissolved in DMSO at doses of 1, 5, and 10 mg/kg for 3 months. A positive control group underwent IP injection of saline followed by daily gavage of saline/DMSO solution. Treatment was stopped 1 week prior to functional assay and euthanasia. MAIN OUTCOME MEASURE: Penile hemodynamics was assessed by electrical stimulation of the cavernous nerves with real-time intracavernous pressure (ICP) measurement. After euthanasia, penile tissue was studied using immunohistochemistry, Western blot, and enzyme-linked immunosorbent assay (ELISA) to assess the nitric oxide-cyclic guanosine monophosphate (NO-cGMP) and TGFß1/Smad2 signaling pathway. RESULTS: Diabetes attenuated ICP response in control animals. Untreated diabetic animals had decreased smooth muscle/collagen ratio and endothelial cell content in the corpora cavernosa; treatment with icariin partially attenuating these effects. Icariin-treated animals also had a significantly greater expression of nicotinamide adenine dinucleotide phosphate-positive nerves and the endothelial cell markers, von Willebrand factor (vWF), and platelet endothelial cell adhesion molecule-1 (PECAM). TGFß1/Smad2 signaling pathway was down-regulated in the penis from icariin-treated models relative to what was observed in negative control animals. CONCLUSION: Icariin treatment preserved penile hemodynamics, smooth muscle and endothelial integrity, and neuronal nitric oxide synthase expression in the penis of diabetic rats. Down-regulation of TGFß1/Smad2 signaling pathway might mediate this effect.

[Effects of icariin and icariside II on eNOS expression and NOS activity in porcine aorta endothelial cells].

OBJECTIVE: To investigate the effects of icariin and icariside II on eNOS expression and NOS activity in endothelial cells and possible mechanisms using EGFR over-expressed porcine aorta endothelial (PAE) cell line. METHODS: The EGFR gene was transfected into PAE cells and genetic stable cell line (PAE-EGFR) was selected. 12.5 µmol/L of icariin and of icariside II were used to treat the PAE and PAE-EGFR cells respectively for 48 h, the eNOS expression in each group was observed. EGF was also used to treat the cells to observe the regulatory effects of icariin and icariside II on NOS activity. The regulatory effects of icariin and icariside II on NOS activity were also observed, and sildenafil was used as a control. RESULTS: Western blot showed that the basic value of eNOS expression was higher in PAE-EGFR group compared with that in PAE group, both of icariin and icariside II increased the eNOS expression in PAE and PAE-EGFR group (P<0.01), and the value of eNOS expression was higher in PAE-EGFR group than that in PAE group. In the PAE-EGFR cell line, the NOS activity reached (15.37 ± 1.49) u/mg when the concentration of icariside II was 10(-8) mol/L, which was 4.66 u/mg more than that in the PAE cell line. When the concentration reached 10(-7), 10(-6) or 10(-5) mol/L, the change of NOS activity in PAE-EGFR group was greater than that in PAE group (P<0.01). icariin also increased the NOS activity in PAE and PAE-EGFR cells, but the activity was 20% lower compared with icariside II group, however, Sildenafil showed no influence on NOS activity. CONCLUSION: Icariin and icariside II may increase the eNOS expression through activating EGF-EGFR pathway in PAE cell, by which endothelial cells function could be regulated and the better effect was noted in icariside II compared to icariin.

[Changes in morphology and steroidogenic function of aged human Leydig cells].

OBJECTIVE: To investigate the changes of morphology and steroidogenic function in aged human Leydig cells and to understand the mechanism of late onset hypogonadism (LOH). METHODS: Ten young and ten aged male subjects were enrolled in this study. AMS (Aging Male's Symptoms) scale was used for symptom evaluation. Testes species with LOH were utilized as the research model. Then the histological changes in testis and ultrastructure of Leydig cells were observed by HE staining and electron microscopy (EM), respectively. The serum total testosterone concentrations were measured by an ELISA kit. The expressions of steroidogenic acute regulatory (StAR) protein and cholesterol-side-chain cleavage enzyme (P450scc) were detected by western blot. RESULTS: The scores of AMS in the aged group were higher than those in the young group with decreased serum testosterone levels (61.25 ± 7.08 vs. 20.75 ± 3.73,P<0.001). And the serum testosterone level of the aged human was lower than that of the young human [(3.12 ± 0.58) µg/L vs. (6.29 ± 1.17) µg/L,P<0.05]. HE staining showed that degenerative changes occurred in the aged human testes. And many swollen mitochondria with mitochondrial cristae that disappeared were found in Leydig cells of the aged human by EM. The serum total testosterone level of the aged human was significantly lower than that of the young group. And the expressions of StAR and P450scc protein in the aged group were significantly lower than those of the young group. CONCLUSION: Mitochondrial swelling and decreased expressions of StAR and P450scc were closely related to the reduced ability of testosterone synthesis in aged males. And the exact mechanism needs further investigation.

Autophagic deficiency is related to steroidogenic decline in aged rat Leydig cells.

Late-onset hypogonadism (LOH) is closely related to secondary androgen deficiency in aged males, but the mechanism remains unclear. In this study, we found that reduced testosterone production in aged rat Leydig cells is associated with decreased autophagic activity. Primary rat Leydig cells and the TM3 mouse Leydig cell line were used to study the effect of autophagic deficiency on Leydig cell testosterone production. In Leydig cells from young and aged rats, treatment with wortmannin, an autophagy inhibitor, inhibited luteinising hormone (LH)-stimulated steroidogenic acute regulatory (StAR) protein expression and decreased testosterone production. In contrast, treatment with rapamycin, an autophagy activator, enhanced LH-stimulated steroidogenesis in Leydig cells from aged, but not young, rats. Intracellular reactive oxygen species (ROS) levels were increased in both young and aged Leydig cells treated with wortmannin but decreased only in aged Leydig cells treated with rapamycin. Furthermore, an increased level of ROS, induced by H(2)O(2), resulted in LH-stimulated steroidogenic inhibition. Finally, knockdown of Beclin 1 decreased LH-stimulated StAR expression and testosterone production in TM3 mouse Leydig cells, which were associated with increased intracellular ROS level. These results suggested that autophagic deficiency is related to steroidogenic decline in aged rat Leydig cells, which might be influenced by intracellular ROS levels.

Current therapeutic strategies for premature ejaculation and future perspectives.

Premature ejaculation (PE) is a common sexual disorder in men that is mediated by disturbances in the peripheral and central nervous systems. Although all pharmaceutical treatments for PE are currently used 'off-label', some novel oral agents and some newer methods of drug administration now provide important relief to PE patients. However, the aetiology of this condition has still not been unified, primarily because of the lack of a standard animal model for basic research and the absence of a widely accepted definition and assessment tool for evidence-based clinical studies in patients with PE. In this review, we focus on the current therapeutic strategies and future treatment perspectives for PE.

[Clinical efficacy and patient satisfaction with penile prosthesis implantation for the treatment of severe erectile dysfunction].

OBJECTIVE: To investigate the clinical efficacy and patient and partner's satisfaction with penile prosthesis implantation (PPI) for treating Chinese patients with severe erectile dysfunction (SED). METHODS: One hundred and sixty-eight SED patients were successfully treated by PPI from July 2000 to June 2010. Of the 146 (86.9%) patients who had been followed up over 6 months post-operation, 36 (24.7%) had been implanted with one piece malleable prosthesis (AMS650) and 110 (75.3%) with three piece inflatable prosthesis (AMS700CXM).All the patients had been followed up by using international index of erectile dysfunction (IIEF5), Quality of Life Score (QOL) for evaluating clinical efficacy and using Visual Analogue Scale (VAS) for evaluating patient and partner's satisfaction and the duration of the follow-up was 6 to 119 months. RESULTS: The mean age of patients was 35.9+/-12.1 years(20 to 75 years), All the operations were successful and sexual intercourse with PPI was performed post 4 to 6 weeks without severe complications like infection and erosion. The prosthesis survive rate and frequent sexual intercourse rate were 98.6% and 87.7% respectively. IIEF5 scores pre and post PPI were 6.3+/- 1.7 and 21.3+/-1.6 respectively,the QOL scores pre and post PPI were 5.1+/-0.9 and 1.5+/-0.5 respectively, and both of them showed significant improvement (P<0.01). As for VAS, the patient and partner's overall satisfaction rates were 92.5% and 90.4% respectively. Moreover, better satisfaction was showed with AMS700CXM as compared with AMS650 (P<0.05) in patients with SED. CONCLUSION: PPI is the safe and effective treatment option for Chinese patients with SED. The AMS700CXM penile prosthesis is better than AMS650 for patients' overall satisfaction.

[Corpus cavernosum-corpus spongiosum shunt plus intracavernous tunneling for the treatment of prolonged ischemic priapism].

OBJECTIVE: To investigate the efficacy and safety of corpus cavernosum-corpus spongiosum shunt (CC-CSS) plus intracavernous tunneling(CC-CSS+ICT) for the treatment of prolonged ischemic priapism (PIP) were investigated. METHODS: Of 21 patients with PIP, 11 (Group A) underwent CC-CSS and 10 (Group B) CC-CSS+ICT surgery. The penile hardness score (PHS) and pain visual analogue score (PVAS) were used to assess the efficacy of the surgery. RESULTS: The erectile functions of the two groups were normal (IIEF5 23.6+/-1.1) before the onset of PIP, and the duration of PIP was (3.4+/-1.3) d. PHS 3.9+/-0.4, and PVAS 8.4+/-0.7. There was no statistical difference between the two groups (P>0.05). On 1, 3 and 5 days after the operation, the PHS and PVAS of Group B decreased significantly than those of Group A (P<0.05). CONCLUSION: CC-CSS+ICT could quickly restore penile detumescence and relieve pain as compared with CC-CSS, which might be a safe and effective method for the treatment of PIP.

Effect of tankyrase 1 on autophagy in the corpus cavernosum smooth muscle cells from ageing rats with erectile dysfunction and its potential mechanism.

This study compared tankyrase 1 expression and autophagy quantity between erectile dysfunction (ED) and non-ED rats' corpus cavernosum smooth muscle cells (CSMCs). This study aslo explored the effect and possible mechanism of tankyrase 1 on autophagy and cell proliferation in ageing ED rats' CSMCs. The intracavernous pressure and mean systemic arterial pressure were measured to investigate erectile function so that eight 24-month-old ED and eight 8-month-old male Wistar rats were chosen respectively. The rat CSMCs were isolated and cultured by enzyme digestion, in which tankyrase 1 expression and autophagy quantity were compared. Tankyrase 1 overexpression was induced with plasmid transfection by Lipofectamine. The effect of tankyrase 1 overexpression on proliferation, autophagy and mTOR pathway in 24-month-old ED rats' CSMCs was measured by the cell growth curve in MTT assay, cell cycle analysis in flow cytometry (FCM), key protein expression in Western blot, autophagy quantity in transmission electron microscopy, monodansylcadaverine staining and GFP-LC3 fluorescence. The primary CSMCs were confirmed by immunofluorescence, and the purity was 99.1% in FCM. Compared with that of 8-month-old rats, tankyrase 1 expression and autophagy quantity significantly decreased in 24-month-old ED rats' primary CSMCs (P < 0.01). Tankyrase 1 overexpression significantly increased the growth rate (P < 0.05) and increased the S phase of cell cycle (P < 0.01). The autophagosome quantity was remarkably increased (P < 0.01), LC3-I/II and Beclin 1 were upregulated (P < 0.01 and P < 0.05), and p-p70S6K (Thr(389)) was downregulated in 24-month-old ED rat CSMCs (P < 0.05). In conclusion, Tankyrase 1 and autophagy decrease in the CSMCs from aging rats with ED, and tankyrase 1 may have a positive effect on proliferation by enhancing autophagy and regulating the mTOR signalling pathway.

Al-Ghorab Shunt plus intracavernous tunneling for prolonged ischemic priapism.

To investigate the efficacy and safety of the corpus cavernosum-corpus spongiosum shunt (Al-Ghorab Shunt) plus intracavernous tunneling (CC-CSS+ICT) for prolonged ischemic priapism (PIP). Twelve patients with PIP were enrolled in this study. The mean age of patients was 38.3 ± 9.2 years old and the mean duration of PIP was 2.8 ± 1.0 days (range, 1.5-4 days). All patients received CC-CSS+ICT for treating PIP. The penile hardness score (PHS) and pain visual analogue score (PVAS) were used to assess the efficacy of the surgery 1, 3, and 5 days after surgery. Color duplex Doppler ultrasonography, International Index of Erectile Function, and quality of life (QOL) were used for evaluating penile morphology, erectile function, QOL, and response to sildenafil treatment. The mean duration of follow up was 21.6 ± 10.1 months. Penile detumescence was successfully restored for all 12 patients postsurgery, with the mean PHS and PVAS significantly decreased compared with that presurgery at different time points (1, 3, 5 days postsurgery; P < .001). The cavernosal arterial blood flow observed with the mean PSV at 1, 3, and 5 days postoperation were 17.79 ± 2.04, 19.14 ± 1.58, and 7.73 ± 2.02 cm/s respectively. All patients suffered from corpus cavernosum fibrosis and erectile dysfunction postoperation; only 2 cases (16.7%) with a short duration of PIP (1.5 days) showed response to sildenafil treatment, and 3 cases (25.0%) with severe fibrosis were satisfied with sexual life after excision of penile corpus cavernosum scar and penile prosthesis implantation. The CC-CSS+ICT could quickly reduce penile rigidity and pain for improving the symptoms of PIP and suggests a safe and effective therapeutic method for PIP.

[Preliminary research on the relationship between inhibitory effect of triamcinolone acetonide on hypertrophic scar and oxygen free radicals in rabbits].

OBJECTIVE: To investigate the relationship between inhibitory effect of triamcinolone acetonide(TA) on hypertrophic scar and oxygen free radicals in rabbits. METHODS: 18 New Zealand rabbits were used. 14 rabbits were selected randomly to construct animal models of hypertrophic scar on the ears. Another 4 rabbits (8 ears) were used as controls. 6 weeks after operation, the hypertrophic scar on the ears were randomly divided to receive intra-lesion injection of TA (n=10), or normal saline (n=10), or nothing (n=8, sham group). 9 weeks after operation, scar specimens were taken for scar thickness measurement, fibroblast counting under microscopy and MDA content detection by spectrophotometric method. RESULTS: (1) 3 weeks after TA treatment, the scar became very thin and soft with a similar color to normal skin and a smooth surface; (2) Histologic study showed the collagen fibers in TA group were reduced markedly and arranged parallelly; (3) Compared with normal skin, the fibroblast density in sham and saline groups increased significantly (P < 0.05), while it was not markedly different in TA group (P > 0.05). There was also no significant difference in scar hypertrophic index between sham and saline groups (P > 0.05), but the scar hypertrophic index was decreased dramatically in TA group (P < 0.05); (4) The MDA content was highest in TA group (P < 0.05), followed by that in sham and saline groups (P < 0.05), while there was no difference between these two groups (P > 0.05). It was lowest in normal control group (P < 0.05). CONCLUSIONS: The oxygen free radicals in the hypertrophic scar can be further increased by local injection of TA.