Short hairpin rna targeting insulin-like growth fator binding protein-3 restores the bioavailability of insulin-like growth factor-1 in diabetic rats.

PURPOSE: To investigate whether intracavernosal injection of short hairpin RNA for IGFBP-3 could improve erectile function in streptozotocin-induced diabetic rats. MATERIALS AND METHODS: After 12 weeks of IGFBP-3 short hairpin RNA injection treatment, intracavernous pressure responses to electrical stimulation of cavernous nerves were evaluated. The expression of IGFBP-3 and IGF-1 at mRNA and protein levels were detected by quantitative real-time PCR analysis and Western blot, respectively. The concentration of cavernous cyclic guanosine monophosphate was detected by enzyme-linked immunosorbent assay. RESULTS: At 12 weeks after intracavernous administration of IGFBP-3 shRNA, the cavernosal pressure was significantly increased in response to the cavernous nerves stimulation compared to the diabetic group (P<0.05). Cavernous IGFBP-3 expression at both mRNA and protein levels was significantly inhibited. At the same time, cavernous IGF-1 expression was significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic group (P<0.01). Cavernous cyclic guanosine monophosphate concentration was significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic group (P<0.01). CONCLUSIONS: Gene transfer of IGFBP-3 shRNA could improve erectile function via the restoration of cavernous IGF-1 bioavailability and an increase of cavernous cGMP concentration in the pathogenesis of erectile dysfunction in streptozotocin-induced diabetic rats.

Short hairpin ribonucleic acid constructs targeting insulin-like growth factor binding protein-3 rehabilitated decreased testosterone concentrations in diabetic rats.

BACKGROUND: The aim of this study was to determine if shRNA constructs targeting insulin-like growth factor binding protein-3 can rehabilitate decreased serum testosterone concentrations in streptozotocin-induced diabetic rats. MATERIAL/METHODS: After 12 weeks of intracavernous administration of IGFBP-3 shRNA, intracavernous pressure responses to electrical stimulation of cavernous nerves were evaluated. The expression of IGFBP-3 at mRNA and protein levels was detected by quantitative real-time PCR analysis and Western blot, respectively. The concentrations of serum testosterone and cavernous cyclic guanosine monophosphate were detected by enzyme-linked immunosorbent assay. RESULTS: After 12 weeks of intracavernous administration of IGFBP-3 shRNA, the cavernosal pressure was significantly increased in response to the cavernous nerves stimulation compared to the diabetic control group (p<0.01). Cavernous IGFBP-3 expression at both mRNA and protein levels was significantly inhibited. Both serum testosterone and cavernous cyclic guanosine monophosphate concentrations were significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic control group (p<0.01). CONCLUSIONS: These results suggest that IGFBP-3 shRNA may rehabilitate erectile function via increases of concentrations of serum testosterone and cavernous cyclic guanosine monophosphate in streptozotocin-induced diabetic rats.

Clinical observation of different minimally invasive surgeries for the treatment of impacted upper ureteral calculi.

OBJECTIVE: To compare the clinical effects of three minimally invasive surgeries on the treatment of impacted upper ureteral calculi. METHODS: 135 patients with impacted upper ureteral calculi were selected and randomly divided into three groups (Group A-C) (n=45), which were treated with transurethral ureteroscopic lithotripsy, minimally invasive percutaneous nephrolithotomy, and retroperitoneal laparoscopic ureterolithotomy respectively. Relevant results of the three groups were compared. RESULTS: The surgery time of Group C was significantly longer than those of Group A and Group B (P < 0.05). The postoperative hospitalization time of Group B was significantly longer than those of Group A and Group C (P < 0.05). 37.78% (17/45) of Group A patients required extracorporeal shock wave lithotripsy, being significantly more than those in Group B (6.67%, 3/45) and Group C (0, 0/45) (P < 0.05). The postoperative calculus clearance rate of Group A (51.11%, 82.22%) was significantly lower than those of Group B (91.11%, 97.78%) and Group C (93.33%, 100%) (P < 0.05). The incidence rates of postoperative complications in Group A-C were 11.11% (5/45), 8.89% (4/45) and 6.67% (3/45) respectively without significant differences (P > 0.05). CONCLUSION: The three surgical methods for impacted upper ureteral calculi should be selected according to practical conditions to improve therapeutic effects and to ensure safe surgery.

Effect of surgical castration on expression of TRPM8 in urogenital tract of male rats.

Trpm8 (melastatin-related transient receptor potential member 8), a member of the transient receptor potential (TRP) superfamily, encoding a cation channel named TRPM8, has been shown to be a primary androgen-responsive gene and play an important role in prostate physiology. To investigate the expression feature of TRPM8 in urogenital tract of male rats, and whether TRPM8 was also regulated by androgen receptor in these organs, male Sprague-Dawley rats were divided into three groups of 35 animals as follows: sham-operated (SHAM), orchidectomized (ORX), orchidectomized plus DHT treatment (O + D). Organs in urogenital tract, including kidney, prostate, seminal vesicle (SV), testis, epididymis and penis, were collected at different post-castration periods. RT-PCR, real-time PCR and Western blotting were used to detect the expression of androgen receptor (AR) and trpm8 in these tissue. As a result, AR and trpm8 can be detected in all these organs at mRNA or/and protein level. The mRNA expression of trpm8 in kidney, prostate, SV and penis decreased 24 or 72 h after castration and kept decreasing in a time-dependant manner. However, treatment of dihydrotestosterone (DHT) could reverse the effect of surgical castration. Collectively, our data provide evidence that TRPM8 and AR were expressed generally in urogenital tract of male rats, and in these organs, expression of trpm8 was regulated by serum androgen.

Clinical Effects of Preoperative K-Line Tilt on Patient Outcomes After Laminoplasty for Cervical Ossification of the Posterior Longitudinal Ligament.

OBJECTIVE: We sought to clarify the effects of the preoperative K-line tilt on cervical sagittal balance and patient outcomes after laminoplasty for cervical ossification of the posterior longitudinal ligament (OPLL). METHODS: A retrospective analysis of 45 patients with OPLL treated by laminoplasty from January 2013 to December 2018 was performed. The radiological parameters included the K-line tilt, C2-C7 sagittal vertical axis, C2-C7 lordosis, T1 slope, and T1 slope minus C2-C7 lordosis. The patient quality of life outcomes were assessed using the neck disability index, Japanese Orthopaedic Association disability scale, and visual analog scale. The patients were classified into 2 groups according to the median preoperative K-line tilt angle (14.1°): the high K-line tilt group (n = 23) and the low K-line tilt group (n = 22). The postoperative cervical alignment changes and patient outcomes were compared and analyzed. RESULTS: The clinical outcomes demonstrated overall improvement at the final follow-up. The C2-C7 lordosis had significantly decreased from 13.5° ± 9.5° preoperatively to 10.2° ± 9.7°. The C2-C7 lordosis was less in the high K-line tilt group than that in the low K-line tilt group. In addition, the high K-line tilt group revealed significantly greater kyphotic changes and a greater loss of cervical lordosis compared with the low K-line tilt group. Finally, the quality of life outcomes and postoperative visual analog scale scores were higher in the high K-line tilt group. CONCLUSIONS: The results of the present study have shown that the parameter K-line tilt is an ideal radiological parameter for predicting the outcomes and determining the need for laminoplasty for cervical ossification of the OPLL. Patients with a higher K-line tilt preoperatively experienced more kyphotic alignment changes and neck pain after laminoplasty.

MIR-138-5P inhibits the progression of prostate cancer by targeting FOXC1.

BACKGROUND: Studies have suggested that micro-RNAs (miRNAs) can function as an oncogene or a tumor suppressor in cancers. However, the role of MIR-138-5P (613394) in prostate cancer (PCa) remains unclear. METHODS: Expression level of MIR-138-5P in PCa cell lines and normal cell line was analyzed with the quantitative real-time PCR method. Cell counting kit-8 assay, colony formation assay, wound-healing assay, and transwell invasion assay were performed to analyze the biological functions of MIR-138-5P. RESULTS: We showed MIR-138-5P expression level was significantly decreased in PCa cell lines compared with the normal cell line. Overexpression of MIR-138-5P inhibits PCa cell proliferation, colony formation, cell migration, and cell invasion in vitro. Mechanistically, we showed Forkhead box C1 (FOXC1, 601090) was a direct target for MIR-138-5P in PCa. We confirmed that overexpression of FOXC1 partially reversed the effects of MIR-138-5P on PCa cell behaviors. CONCLUSIONS: Collectively, we showed that MIR-138-5P functions as a tumor suppressor gene in PCa via targeting FOXC1.

[Vas-to-Epididymis antidromic injection of 30% ethanol reduces sperm motility in rats].

OBJECTIVE: To investigate the reduction of sperm motility in rats induced by vas-to-epididymis antidromic injection of 30% ethanol and its mechanism. METHODS: Forty male adult Sprague-Dawley rats were randomized into 3 groups: bilateral vas injection (n = 15) , sham operation control (n = 15) and normal (n = 10). An aliquot of 0.5 ml of 30% ethanol was injected from vas to epididymis bilaterally. After 1 month, all the rats'vasa and epididymides were ablated for studies of the sperm motility, construction changes of the vas and contents of IL-6, IFN-gamma and carnitine of the epididymis. RESULTS: There was markedly significant difference in sperm motility in the injection group (P < 0.01). The number of sperms in the bilateral vas injection group was 31, while in the sham operation control and normal groups was 64 and 68, respectively. The contents of IL-6 and IFN-gamma increased, and the carnitine reduced significantly (P < 0.05). However, no significant differences were noted between the control and the normal groups (P > 0.05). The contents of IL-6, IFN-gamma and carnitine in the bilateral vas injection group were 772.7 pg/ml, 350.7 pg/ml and 491.1 mol/L. But the same indexes in the sham operation and normal groups were 308.5 pg/ml, 172. 2 pg/ml and 664. 6 mol/L and 287. 8 pg/ml, 163. 8 pg/ml and 605.5 mol/L. CONCLUSION: The antidromic injection of ethanol from vas to epididymis can not only interfere the environment for sperm maturation but also activate the immunologic cells that secrete many cytokines (CK) in the genital system. All the factors can induce the reduction of sperm motility.

[Antifertility effect of 30% ethanol retro-injection into rat vas deferens].

OBJECTIVE: To explore the antifertility effect and safety of 30% ethanol retro-injection into the vas deferens of the rat. METHODS: Thirty Sprague-Dawley male rats, 3 m of age and (200 +/- 20) g in weight, were equally randomized into an experimental group and a control group. The former received 30% ethanol (0.5 ml) and the latter 0.9% sodium chloride (0.5 ml), both retro-injected into the vas deferens. Pregnancy rates were obtained through pregnancy tests with 60 Sprague-Dawley female adult rats 1.5 m and 3 m after the injection. All the male rats were sacrificed three months later, and tests were done for the rates of sperm motility and deformity as well as for the apoptosis of spermatogenic cells with TUNEL. RESULTS: The 1.5 m pregnancy rate was 0 and the 3 m sperm motility and pregnancy rates were (0.32 +/- 1.12)% and (0.58 +/- 1.27)%, significantly decreased (P < 0.05) as compared with those of the control group, which were (80.62 +/- 2.68)%, (70.68 +/- 1.62)% and (86.62 +/- 1.68)%, respectively. While the 3 m sperm deformity rate in the experimental group was (78.26 +/- 1.08)%, increased significantly (P < 0.05), and the apoptosis index (AI) of spermatogenic cells was (7.63 +/- 1.16)% as compared with (5.62 +/- 1.32)% of the control group, with no significant difference between the two groups (P > 0.05). CONCLUSION: Retro-injection of 30% ethanol into the vas deferens of the rat produces significant antifertility effect on rats, but has no significant influence on their spermatogenic cells.

Short hairpin ribonucleic acid constructs targeting insulin-like growth factor binding protein-3 ameliorates diabetes mellitus-related erectile dysfunction in rats.

OBJECTIVE: To investigate the expression of insulin-like growth factor binding protein-3 (IGFBP-3) in penile cavernous of streptozotocin (STZ)-induced DM rats and whether downregulation of IGFBP-3 by intracavernosal injection of short hairpin ribonucleic acid (shRNA) targeting IGFBP-3 could improve the erectile function in DM rats. MATERIALS AND METHODS: Diabetes was induced in rats by intraperitoneal injection of STZ, and the expression of IGFBP-3 in the penile tissue of adult normal and DM male rats was assayed using reverse transcriptase-polymerase chain reaction and Western blot. Next, shRNA-targeting IGFBP-3 and a scramble sequence were injected into the penile corpora cavernosa of DM rats. At 12 weeks after shRNA-IGFBP-3 administration, the intracavernous pressure in response to electrical stimulation of the cavernous nerves was evaluated. The expression of IGFBP-3 was assayed by Western blot. The concentration of cyclic guanosine monophosphate in the corpus cavernosum was assayed by enzyme-linked immunosorbent assay. RESULTS: At 12 week after intraperitoneal administration of STZ, IGFBP-3 expression had increased in the penis of the DM rat (P <.05) compared with that of the normal control rats. Among the DM rats, IGFBP-3 expression at the messenger RNA and protein level was significantly inhibited 12 weeks after intracavernous administration of IGFBP-3 shRNA (P <.01). At 12 weeks after shRNA-IGFBP-3 injection, intracavernosal pressure was significantly increased in response to cavernous nerve stimulation (P <.05), and an increase in the concentration of cyclic guanosine monophosphate in the corpus cavernous tissue (P <.01) was detected compared with the "randomer" shRNA treatment group. CONCLUSION: Gene transfer of shRNA-IGFBP-3 could improve erectile function in STZ-induced DM rats by an increase in the cyclic guanosine monophosphate concentration in cavernous tissue.

Increased expression of insulin-like growth factor-binding protein-3 is implicated in erectile dysfunction in two-kidney one-clip hypertensive rats after propranolol treatment.

This study aimed to investigate the role of insulin-like growth factor-binding protein-3 (IGFBP-3) in erectile dysfunction (ED) in two-kidney one-clip (2K-1C) hypertensive rats treated with the ß-blocking agent propranolol. Adult male Wistar rats were randomly divided into three groups: a normal control group, a hypertensive control group and a propranolol treatment group (n=9). After 4 weeks of propranolol treatment, intracavernous pressure (ICP) responses to electrical stimulation of the cavernous nerves were evaluated. The expression of IGFBP-3 and insulin-like growth factor-1 (IGF-1) mRNA and protein in the rat cavernous tissue were detected by quantitative real-time PCR and Western blot, respectively. The concentration of cyclic guanosine monophosphate (cGMP) in the cavernous tissue was determined by enzyme-linked immunosorbent assay (ELISA). Cavernosal pressure in response to cavernous nerve stimulation was decreased 4 weeks after propranolol treatment (P<0.01, compared to the hypertensive control group). IGFBP-3 mRNA and protein expression was increased in the propranolol treatment group compared to the hypertensive control group (P<0.01), whereas IGF-1 expression was decreased in the propranolol treatment group compared to the hypertensive control group (P<0.01). In addition, cavernous cGMP concentration was decreased in the propranolol treatment group compared to the hypertensive control group (P<0.01). Taken together, these results suggest that the upregulation of IGFBP-3 may play a role in the development of ED in hypertensive rats.

Short hairpin RNA targeting insulin-like growth factor binding protein-3 restores the bioavailability of insulin-like growth factor-1 in diabetic rats

ABSTRACT Purpose To investigate whether intracavernosal injection of short hairpin RNA for IGFBP-3 could improve erectile function in streptozotocin-induced diabetic rats. Materials and methods After 12 weeks of IGFBP-3 short hairpin RNA injection treatment, intracavernous pressure responses to electrical stimulation of cavernous nerves were evaluated. The expression of IGFBP-3 and IGF-1 at mRNA and protein levels were detected by quantitative real-time PCR analysis and Western blot, respectively. The concentration of cavernous cyclic guanosine monophosphate was detected by enzyme-linked immunosorbent assay. Results At 12 weeks after intracavernous administration of IGFBP-3 shRNA, the cavernosal pressure was significantly increased in response to the cavernous nerves stimulation compared to the diabetic group (P<0.05). Cavernous IGFBP-3 expression at both mRNA and protein levels was significantly inhibited. At the same time, cavernous IGF-1 expression was significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic group (P<0.01). Cavernous cyclic guanosine monophosphate concentration was significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic group (P<0.01). Conclusions Gene transfer of IGFBP-3 shRNA could improve erectile function via the restoration of cavernous IGF-1 bioavailability and an increase of cavernous cGMP concentration in the pathogenesis of erectile dysfunction in streptozotocin-induced diabetic rats.