Suppression of Tescalcin inhibits growth and metastasis in renal cell carcinoma via downregulating NHE1 and NF-kB signaling.

BACKGROUND: Renal cell carcinoma (RCC) is the most common form of kidney cancer. Recent studies reported that Tescalcin was overexpressed in various tumor types. However, the status of Tescalcin protein expression in RCC and its biological function is uncertain. This study was designed to investigate the expression of Tescalcin in human RCC and its biological function. METHODS: shRNA transfection was performed to abrogates the expression of Tescalcin. Quantitative real time PCR and western blotting assays were used to determine mRNA and protein expression levels, respectively. The cell viability was analyzed by MTT and colony formation. Cell flow cytometry was used to assess pHi value and cell apoptosis. Cell invasive and migratory ability was measured with modified Boyden chamber assay. Xenograft model was setup to evaluate tumor growth. RESULTS: Tescalcin was overexpressed in RCC tissues compared with matched normal tissues. It was also overexpressed in RCC cell lines relative that of normal cells. Suppression Tescalcin with specific shRNA resulted in the inhibition of cell proliferation, migration, invasion and apoptosis of RCC cells. Additionally, silencing of Tescalcin also caused the inhibition of the tumor growth in nude mice. Mechanistic study showed that Tescalcin regulated cell proliferation, migration and invasion via NHE1/pHi axis as well as AKT/NF-κB signaling pathway. CONCLUSIONS: These findings demonstrate that atopic expression of Tescalcin facilitates the survival, migration and invasion of RCC cells via NHE1/pHi axis as well as AKT/ NF-κB signaling pathway, providing new perspectives for the future study of Tescalcin as a therapeutic target for RCC.

[Causes of orchiectomy: An analysis of 291 cases].

OBJECTIVE: To study the causes of orchiectomy in different age groups. METHODS: We retrospectively reviewed the clinical data about 291 cases of orchiectomy performed between March 1993 and October 2014 and analyzed the causes of surgery and their distribution in different age groups. RESULTS: The main causes of orchiectomy were testicular torsion (45.8%), cryptorchidism (32.5%) and testicular tumor (16.9%) in the patients aged 0-25 years, testicular tumor (42.4%), cryptorchidism (25.9%) and tuberculosis (10.6%) in those aged 26-50 years. Prostate cancer was the leading cause in those aged 51-75 years (77.6%) or older (84.0%)), and testicular tumor was another cause in the 51-75 years old men (10.2%). Prostate cancer, testicular tumor, cryptorchidism, and testicular torsion were the first four causes of orchiectomy between 1993 and 2009. From 2010 to 2014, however, testicular tumor rose to the top while prostate cancer dropped to the fourth place. CONCLUSION: The causes of orchiectomy vary in different age groups. The proportion of castration for prostate cancer patients significantly reduced in the past five years, which might be attributed to the improvement of comprehensive health care service.

[Low-dose tadalafil for erectile dysfunction following pelvic fracture-induced urethral injury: clinical observation of 42 cases].

OBJECTIVE: To evaluate the efficacy of daily low-dose tadalafil in the treatment of erectile dysfunction (ED) following pelvic fracture-induced urethral injury (PFUI). METHODS: Based on the length of time between pelvic fracture and the patients' clinic visit, we divided 42 PFUI-caused ED patients into groups A (< 1 month), B (6-24 months) and C (> 24 months). We treated them with tadalafil at 5 mg daily for 12 weeks consecutively, followed by evaluation of the therapeutic effect using IIEF-5 questionnaire and Sexual Encounter Profile (SEP) diaries. RESULTS: Thirty-four patients (83.3%) completed the investigation and all responded well to tadalafil medication. Group A showed significant differences from B and C in the increase of IIEF-5 scores and the positive rate of SEP. CONCLUSION: Daily low-dose tadalafil helps penile rehabilitation in ED patients following PFUI, and the earlier the medication is initiated, the better the effect will be.

Association between reversal of multidrug resistance by methyl jasmonate and P-glycoprotein ATPase activity in hepatocellular carcinoma.

OBJECTIVE: To study the effects of methyl jasmonate on multidrug resistance in a mouse model of hepatocellular carcinoma. METHODS: Multidrug resistant H22 (H22/FAP) hepatocellular carcinoma cells were produced in vitro by continuous exposure to increasing doses of doxorubicin, cisplatin and 5-fluorouracil (FAP regimen). Cell toxicity was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolum bromide (MTT) assay. Survival time was calculated for BALB/c mice that received intraperitoneal injections of H22/FAP cells followed by treatment with methyl jasmonate or verapamil in combination with FAP for 7 days. Adenosine triphosphate (ATP) hydrolysis was used to measure the activity of permeability-glycoprotein (P-gp) ATPase activity in plasma membranes. RESULTS: The MTT assay showed that methyl jasmonate significantly enhanced the cytotoxicity of the FAP regimen in multidrug resistant H22/FAP cells. Methyl jasmonate (10 mg/kg and 5 mg/kg) combined with FAP significantly increased survival time in BALB/c mice by 44.25% and 48.01%, respectively, compared with FAP. Methyl jasmonate increased P-gp ATPase activity. CONCLUSION: The combined use of methyl jasmonate and the FAP regimen might be a novel strategy for overcoming the multidrug resistance often observed in hepatocellular carcinoma.

[Distribution of HoxA13 in the prepuce and urethral plate in different types of hypospadias and its implication].

OBJECTIVE: To investigate the role of the HoxA13 gene from the HOX family in the development of hypospadias by detecting the transcription and expression of HoxA13 in the prepuce and urethral plate of hypospadias patients. METHODS: We collected the tissues from the prepuce and urethral plate of 30 hypospadias patients aged 3.3 - 11.6 years, the prepuce of 10 phimosis children aged 3.1 - 10.4 years and the urethra of 10 penile carcinoma patients aged 48.1 - 75.6 years with no urethral involvement, the latter 20 taken as controls. We divided the tissue samples into a distal, an intermedial, a proximal and a control group, and detected the expressions of HoxA13 mRNA and protein in different groups by RT-PCR and immunohistochemistry. RESULTS: RT-PCR showed that the HoxA13 mRNA expressions in the prepuce and urethral plate were significantly higher in the control group (1.409 +/- 0.441 and 1.270 +/- 0.209) than in the intermedial (0.848 +/- 0.338 and 0.684 +/- 0.228) and proximal group (0.497 +/- 0.218 and 0.464 +/- 0.164) (P < 0.05 or P < 0.01), and so were they in the distal (1.071 +/- 0.342 and 1.054 +/- 0.189) than in the proximal group (P < 0.05). Immunohistochemistry revealed that the HoxA13 protein expressions in the prepuce and urethral plate were remarkably higher in the control group (12 050 +/- 4 112 and 13 420 +/- 2 636) than in the intermedial (5 217 +/- 1 993 and 5 238 +/- 3 065) and proximal group (2 095 +/- 1 591 and 2 238 +/- 2 217) (P < 0.05 or P < 0.01), and so were they in the distal (8 223 +/- 3 212 and 10 450 +/- 2 123) than in the proximal group (P < 0.05). CONCLUSION: The transcription and expression of HoxA13 in the prepuce and urethral plate of hypospadias patients are closely related with the abnormal position of the urethral meatus, and their abnormal expressions may affect the development and formation of the urethra.

[Transfection of iNOS suppresses the growth of androgen-independent prostate cancer DU145 cells].

OBJECTIVE: To observe the effect of the inducible nitric oxide synthase (iNOS) gene on androgen-independent prostate cancer DU145 cells in vitro. METHODS: The iNOS gene was transfected into androgen-independent prostate cancer DU145 cells. The positive cells were selected as the transfected group for amplification, and an empty vector (EV) group and a control group were also set. The mRNA transcription of iNOS was analyzed by RT-PCR. The morphological changes of the cells were observed, the effect of iNOS transfection on the cell growth determined using the MTB method, and the apoptosis of DU145 cells detected by flow cytometry, followed by analysis of the effect of NOS inhibitors on the transfected cells. RESULTS: DU145 cells transfected with iNOS secreted significantly more nitric oxide ([272.50 +/- 15.82] micromol/L) than those of the EV and control groups ([122.00 +/- 18.93] micromol/L and [121.00 +/- 6.98] micromol/L) (P < 0.05). The rate of cell apoptosis was markedly enhanced in the transfected group as compared with the EV and control groups ([42.78 +/- 2.01]% vs [30.65 +/- 1.46]% and [28.96 +/- 1.50]%, P < 0.05). MTP test indicated a slower growth of the DU145 cells in the former than in the latter two (P < 0.05). NOS inhibitors enhanced their growth, but with no significance (P > 0.05). CONCLUSION: DU145 cells transfected with iNOS could secrete high-concentration nitric oxide, induce cell apoptosis, and suppress cell proliferation, which may provide a potentially effective gene therapy for advanced androgen-independent prostate cancer.

[Effects of tadalafil on erectile dysfunction: on-demand versus once-daily dosing].

OBJECTIVE: To evaluate the effects of tadalafil administered on demand or once a day in the treatment of erectile dysfunction (ED). METHODS: We randomly assigned 61 ED patients to three groups to receive tadalafil on demand, at 5 mg once daily, and at 10 mg once daily, respectively. After 42 days of medication, we compared the therapeutic effects among different groups using the patients' sexual encounter profile (SEP) diaries, detected the adverse reactions and assessed the safety of tadalafil. RESULTS: Fifty-three (86.7%) of the patients completed the investigation, and all responded well to tadalafil medication, with a significantly improved success rate of sexual intercourse and a low rate of mild adverse effects. The mean positive rates of SEP were basically similar between the on-demand and once-daily groups. CONCLUSION: There are no significant differences in the improvement of penile erection and sexual satisfaction of ED patients treated by on-demand and once-daily administration of tadalafil.

[Transverse preputial island flap technique (Duckett's procedure) for hypospadias repair: a report of 356 cases].

OBJECTIVE: To evaluate the effect of the transverse preputial island flap technique (Duckett's procedure) for hypospadias repair. METHODS: A total of 356 patients with hypospadias were treated by Duckett's procedure from March 1995 to December 2010, of whom 324 (91.0%) were younger than 14 years. The length of urethra repair ranged from 1.5 to 10 cm. RESULTS: The total success rate of Duckett's procedure was 91.0%. Urethra fistula occurred in 30 cases, external orifice stricture in 1, and urethral anastomosis stricture in another. There were no significant differences in the rate of complications among either different age groups or different surgical times (P > 0.05). CONCLUSION: Duckett's procedure remains the first choice for the one-stage repair of hypospadias, especially applicable to hypospadias with chordee.

SPAG4L, a novel nuclear envelope protein involved in the meiotic stage of spermatogenesis.

SUN domain-containing proteins belong to a novel protein family. To date, several members--SUN1, SUN2, SUN3, and SPAG4--have been identified as nuclear envelope (NE) proteins. In this study, we sought to characterize and define the potential function of SPAG4L, a newly identified SUN protein. Using bioinformatic analysis, we found that SPAG4L contained a conserved SUN domain in the C-terminal. Subcellular localization analysis indicated that the expression of green fluorescent protein-labeled full-length SPAG4L was localized to the NE and the endoplasmic reticulum (ER). Deletion analysis revealed that the transmembrane region and the coiled-coil domain, but not the SUN domain, were required for localization of SPAG4L to the NE and ER. Subsequently, we confirmed that the human testes expressed endogenous SPAG4L as a 43-kDa protein. Further studies revealed that mouse Spag4L colocalized with the NE marker Lamin B1 and the ER marker PDI in isolated mouse spermatocytes. In addition, the expression of Spag4L was observed in meiosis I and II stages, suggesting that Spag4L may be involved in NE reconstitution and nuclear migration occurring during the process of spermatocyte division. Together, the findings indicate that SPAG4L, a new NE protein, may play an important role in the meiotic stage of spermatogenesis.

[Causes, location and management of BPH surgery-related urethral stricture].

OBJECTIVE: To analyze the possible causes of BPH surgery-related urethral stricture and summarize the experience in its clinical management. METHODS: The clinical data of 37 cases of BPH surgery-related urethral stricture were analyzed retrospectively. The patients averaged 68.5 years in age, of whom 12 had the history of open surgery, and 25 transurethral surgery. Anterior urethral stricture was found in 6 cases, and posterior urethra in stricture 31. Thirty-five cases were treated by holmium laser urethrotomy (HLU) or a combination of HLU with transurethral resection of the scar in the stenotic segment, and the results were evaluated based on the maximum flow rate (Qmax). RESULTS: Two cases of full penile urethral stricture were treated by urinary diversion, and all the rest by urethral stricture surgery. Catheters were indwelt in 4 cases of urethral stenosis for 5-7 days post-operatively, and smooth urination was achieved after their removal, with a Qmax of > 15 ml/s. Another 31 cases of membranous urethral stricture received catheter indwelling of 2 -4 weeks and were followed up for 1 - 21 (mean 11. 5) months, of whom 23 experienced satisfactory voiding, with an average Qmax of 14.3 ml/s, and the other 8 poor voiding, with a Qmax of < 8 ml/s. CONCLUSION: HLU or a combination of HLU with transurethral resection of the scar in the stenotic segment is an effective method for the treatment of BPH surgery-related urethral stricture. And conscientiously performed BPH surgery can reduce the incidence of urethral stricture.

[Prokaryotic expression and purification of SPAG4L, a novel human testis gene].

OBJECTIVE: To express SPAG4L, a novel human testis gene in E. coli and purify it's fusion protein. METHODS: The fragment encoding SPAG4L126-379 was amplified by RT-PCR and the PCR products were cloned into PUCm-T vectors. After digestion by EcoR I and Hind III, the fragment was subcloned into PQE-30, a prokaryotic expression vector with 6×His tag. The recombinant plasmid PQE-30-SPAG4L was sequenced and transformed into E.coli M15. The expression of his-tagged fusion protein was induced by IPTG. The fusion protein was identified by Western blotting and purified using Ni-NTA magnetic agarose beads. RESULTS: The recombinant plasmid PQE-30-SPAG4L was constructed successfully and expressed in E.coli M15. The fusion protein SPAG4Lwith 6×his-tag was confirmed by Western blotting. The micro-scale purification system of 6×His-tagged SPAG4Lprotein was established and purified fusion protein was obtained. CONCLUSION: The recombinant plasmid PQE-30-SPAG4L can be expressed in vitro and used for studying the biological function of SPAG4L in spermatogenesis.

[Urethral dilatation with the renal sheath dilator under the ureteroscope to treat male urethrostenosis].

OBJECTIVE: To explore the efficacy of urethral dilatation with the renal sheath dilator under the ureteroscope in the treatment of male patients with urethrostenosis. METHODS: Eighteen male patients with urethrostenosis underwent urethral dilatation with the renal sheath dilator. Under the ureteroscope, a zebra-guide wire was inserted through the stenosed urethra into the bladder and the stenosis was gradually dilated with the renal sheath dilator, followed by placing a Foley catheter of proper size for 1-4 weeks. For children, the renal sheath dilator was selected according to their age, while for adults, metal dilators (> or = F20) were used following dilatation with the F18 renal sheath dilator. All the patients were followed up for 6-24 months. RESULTS: The operation was successfully performed in all the 18 cases, with no urethral false passage, urethral perforation or urethra tearing. Sixteen of the patients were cured, and the other 2 received urethroplasty for stenosis recurrence. The maximum flow rate was increased to 13.6-30.2 (18.1 +/- 3.5) mL/s after the operation. CONCLUSION: Urethral dilatation with the renal sheath dilator under the ureteroscope is a simple, safe and effective method for the treatment of urethrostenosis.

[Molecular cloning of a novel human testis-specific gene TDRG1].

OBJECTIVE: To clone a novel human testis-specific gene TDRG1. METHODS: A new contig of expression sequence tags (ESTs) Hs.180197 was identified from the testis libraries using digital differential display (DDD) to screen the novel human testis-specific gene. To validate the use of bioinformatics approaches in gene discovery, the ESTs Hs.180197, which was predicted to be testis specific, was chosen for further study. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on different normal tissues to identify the expression of Hs.180197 in human testis. Using bioinformatics methods and IMAGE cloning of this contig, the full-length cDNA sequence of the noval human gene was cloned. RESULTS: This novel gene was 1197 bp in length, located in chromosome 6p21.1-p21.2. The sequence of the open reading frame was 504-806 bp, as confirmed by RT-PCR and sequencing in human testis. The cDNA encodes a novel protein of 100 amino acids with a theoretical molecular weight of 10 000 and isoelectric point of 6.81. The sequence shares no significant homology with any known protein in the databases. Semi-quantitative RT-PCR analysis of multiple tissues further showed that the novel gene was expressed specifically in adult human testis. Considering a possible relation of this novel gene with the function of human testis, we named this new gene TDRG1 (testis development related gene 1, GenBank accession number: DQ168992). CONCLUSION: DDD combined with laboratory validation is an efficient method for identifying new human functional genes.

[mRNA and protein expression of haptoglobin in lesions of condyloma acuminatum].

OBJECTIVE: To investigate the expression of haptoglobin in the lesions of condyloma acuminatum (CA) at the mRNA and protein level, and to explore its role in the pathogenesis of CA. METHODS: The expressions of haptoglobin protein and mRNA in the skin tissues of 30 patients with CA and 20 normal controls were detected by immunohistochemistry(IHC), Western blot, and hybridization in situ. RESULTS: The in situ hybridization study showed that haptoglobin mRNA was expressed in the epidermal cells in the lesions of CA. The distribution of haptoglobin mRNA expression in the lesions of CA was similar to that of the normal controls, and the expression of haptoglobin mRNA in CA was higher than that of the normal controls. There was a significant difference in the positive expression of haptoglobin mRNA between the CA group and the control group (P<0.05). The immunohistochemical study showed that haptoglobin protein was expressed in the whole layers of epidermal keratinocytes in the lesions of CA at a high level and stronger staining was seen in the stratum basale and stratum spinosum. Haptoglobin protein was expressed predominantly in the stratum basale in normal skin tissues, while weak staining was seen below the stratum spinosum.There was a significant difference in the mean gray value between the CA group and control group (P<0.05). Western blot showed that the haptoglobin expression in CA lesions significantly increased compared with the normal skins (P<0.05). CONCLUSION: The expression of haptoglobin mRNA in the CA lesions obviously increases and the epidermal cells in the CA lesions are able to synthesize haptoglobin protein. Haptoglobin in the CA lesions may involve in the local immunity escape by preventing Langerhans cell functional maturation and inhibiting the immunocompetence of keratinocyte.

[Effect of androgen on nitric oxiode synthase activity in the kidney of rats with ischemic injury].

OBJECTIVE: To investigate the effect of androgen on nitric oxide synthase activity and kidney function in the kidney of rats with ischemic injury. METHODS: Fourty-eight adult healthy male SD rats were randomly divided into the renal ischemia group (n = 24 ) and the orchidectomized group with renal ischemia (n = 24). The two groups were randomly divided into 4 groups by the survival time: 0, 1, 3 and 7 days. NADPH diaphorase histochemistry was used to measure the nitric oxide synthase activity of the kidney with ischemic injury in the orchidectomized group and the control group. The image analysis technology system was used to quantify the NOS. BUN and BCr in the plasma were detected when the rats were executed. RESULTS: The NOS histochemical staining of the kidney was mainly distributed in the renal tubule, macula densa and low in the renal glomerular. The NOS activity of macula densa significantly decreased in the orchidectomized group compared with that in the control group (P < 0.05) . BUN and Bcr have no significant difference on 0, 1 and 7 days (P > 0.05) , but on the third day the data markedly increased (P < 0.05). CONCLUSION: Androgen can adjust the blood flux of glomerular by regulating the NOS activity of macula densa, and affect the kidney function.

[Uroflowmetry and its influence factors in benign prostate hyperplasia patients].

OBJECTIVE: To determine the relationship between uroflowmetry and age, the course of disease, premicturition volume, transition zone index and proportion of stroma-to-epithelium in benign prostate hyperplasia (BPH) patients. METHODS: Eighty-nine BPH patients in our hospital from 2000 to 2003 were evaluated. With the CMM3 pathology image analysis system, transrectal ultrasound and Dantec 2000 urodynamic instrument, the value of influence factors was determined. A linear regression was applied to analyze all the data by SPSS software. RESULTS: The flow rate was correlated to premicturition volume ( r = 0. 477, P < 0.01) and proportion of stroma-to-epithelium significantly ( r = - 0.437, P < 0.05) , but was not correlated to the age, the course of disease and transition zone index significantly. The parameter of flow rate/premicturition volume had no difference in 3 micturitions in all patients. CONCLUSION: The parameter (flow rate/premicturition volume)could be used to evaluate the micturition status of the BPH patients whose bladder volume is less than 200 ml. We should pay more attention to receptor blockers when we treat BPH patients.

[Erectile dysfunction induced by pelvic fracture urethral injury].

OBJECTIVE: To explore the aetiology, diagnosis,and treatment of erectile dysfunction (ED) after a pelvic fracture urethral injury. METHODS: Rigiscan and Duplex ultrasound were used to determine the causes of ED induced by pelvic fracture urethral injury in 29 patients and the therapeutic means were also analyzed. RESULTS: Of the 29 patients, 1 was psychological, 28 were physical, 7 vascular, and 15 neurological. Rigiscan and Duplex ultrasound revealed the cause of erectile failure as neurogenic in 15. Twenty-three cases were effective by the therapy. The injection with vasoactive agents (prostagland in E1) was successful in 15, Viagra in 2, Befar in 5, and revascularization of epigastric artery-dorsal artery anastomosis in 1. CONCLUSION: Disruption of the cavernous nerves lateral to the prostatomembranous urethra is the most likely cause of ED in this injury. Intracavernous injection of prostaglandin E1 is believed to be safe and effective for ED.