Weimin Yang

Calcium oxalate monohydrate crystals stimulate monocyte chemoattractant protein-1 and transforming growth factor β1 expression in human renal epithelial cells.

Crystal-cell interactions play a key role in the formation of kidney stones. Few studies have referred to the role of monocyte chemoattractant protein-1 (MCP-1) and transforming growth factor β1 (TGFβ1) in kidney stone formation. Recently, a genome-wide analysis of genes related to kidney stone formation and eliminiation in mice indicated that MCP-1 and TGFβ1 are involved in nephrolithiasis. In this study, in order to verify whether MCP-1 and TGFβ1 are involved in the process of crystal-cell interactions in vitro, we observed the effects of calcium oxalate monohydrate (COM) on MCP-1 and TGFβ1 expression in cultured HK-2 cells. HK-2 cells were treated with different concentrations of COM, and a group of untreated cells served as the control. The expression of MCP-1 and TGFβ1 was detected by western blot analysis after treatments with different COM concentrations (300, 500, 700 and 900 µg/ml) for different times (3, 6, 12 and 24 h). We found that the expression of MCP-1 was upregulated by COM treatment in a dose-dependent manner, and was increased initially at the first 6 h of treatment, then slightly decreased over time. Also, COM treatment resulted in a dose-dependent increase in TGFβ1 expression, and the expression levels peaked at 12 h. This study demonstrates that COM stimulates the expression of MCP-1 and TGFβ1 in renal epithelial cells.

Clinical assessment and genomic landscape of a consanguineous family with three Kallmann syndrome descendants.

Although some genes that cause Kallmann syndrome (KS) have been identified by traditional linkage analysis and candidate gene techniques, the syndromes molecular etiology in the majority of patients remains poorly understood. In this paper, we present the clinical assessments of a consanguineous Han Chinese family with three KS descendants. To understand the molecular etiology of KS from a genome-wide perspective, we investigated the genome-wide profile of structural variation in this family using the Affymetrix Genome-Wide Human SNP Array 6.0 platform. The results revealed that the three affected individuals had common copy number variants (microdeletions) on chromosomes 1p21.1, 2q32.2, 8q21.13, 14q21.2 and Xp22.31. Moreover, the copy number variants on Xp22.31 were located in the intron of KAL1, which causes X-linked KS. Two PCR assays were performed on these regions to validate the results obtained using the chips. In addition, genomic microdeletions in this region were verified in one of 29 Han Chinese sporadic KS cases and one of four other family cases, but not in 26 Han Chinese sporadic normosmic idiopathic hypogonadotropic hypogonadism cases and 100 unrelated Han Chinese normal controls. Our results provide a novel insight into the relative contributions of certain copy number variants to KSs molecular etiology and generate a list of interesting candidate regions for further studies.

Audiovisual Sexual Stimulation and RigiScan Test for the Diagnosis of Erectile Dysfunction

Background: Currently available evaluation criteria for penile tumescence and rigidity have been fraught with controversy. In this study, we sought to establish normative Chinese evaluation criteria for penile tumescence and rigidity by utilizing audiovisual sexual stimulation and RigiScan™ test (AVSS-Rigiscan test) with the administration of phosphodiesterase-5 inhibitor. Methods: A total of 1169 patients (aged 18–67 years) complained of erectile dysfunction (ED) underwent AVSS-RigiScan test with the administration of phosphodiesterase-5 inhibitor. A total of 1078 patients whose final etiological diagnosis was accurate by means of history, endocrine, vascular, and neurological diagnosis, International Index of Erectile Function 5 questionnaire, and erection hardness score were included in the research. Logistic regression model and receiver operating characteristic curve analysis were performed to determine the cutoff value of the RigiScan™ data. Then, the multivariable logistic analysis was used in the selected variables. Results: A normal result is defined as one erection with basal rigidity over 60% sustained for at least 8.75 min, average event rigidity of tip at least 43.5% and base at least 50.5%, average maximum rigidity of tip at least 62.5% and base at least 67.5%, △tumescence (increase of tumescence or maximum−minimum tumescence) of tip at least 1.75 cm and base at least 1.95 cm, total tumescence time at least 29.75 min, and times of total tumescence at least once. Most importantly, basal rigidity over 60% sustained for at least 8.75 min, average event rigidity of tip at least 43.5%, and base at least 50.5% would be the new normative Chinese evaluation criteria for penile tumescence and rigidity. By multivariable logistic regression analysis, six significant RigiScan™ parameters including times of total tumescence, duration of erectile episodes over 60%, average event rigidity of tip, △tumescence of tip, average event rigidity of base, and △tumescence of base contribute to the risk model of ED. In logistic regression equation, predict value P < 0.303 was considered as psychogenic ED. The sensitivity and specificity of the AVSS-RigiScan test with the administration of phosphodiesterase-5 inhibitor in discriminating psychogenic from organic ED was 87.7% and 93.4%, respectively. Conclusions: This study suggests that AVSS-RigiScan test with oral phosphodiesterase-5 inhibitors can objectively assess penile tumescence and rigidity and seems to be a better modality in differentiating psychogenic from organic ED. However, due to the limited sample size, bias cannot be totally excluded.

AB061. Clinical analysis of small cell carcinoma of the bladder: 9 cases report and literature review

Objective To present our experience with nine patients with small cell carcinoma of the bladder (SCCB) who were treated with different modalities and review the literature for patients with SCCB who have been reported in 56 literatures. SCCB is a rare, highly aggressive tumor that presents in an advanced stage and has a propensity for early metastasis. Hematuria is the main clinical manifestations. Surgery, chemotherapy, and radiotherapy, either alone or as part of combined therapy have been used for treatment. Methods We retrospectively evaluated nine patients with SCCB by medical record review between February 1980 and January 2014 at Tongji Hospital of Huazhong University of Science and Technology. In order to better understand the clinical features of SCCB, 56 literatures were concerned (from January 1979 to March 2014). The general characteristics, clinical manifestation, the pathological and immunohistochemical characteristics, treatment options and prognostication in those eligible manuscripts are analyzed, a retrospective analysis is performed. Results All the nine cases in Tongji hospital were successfully operated and tissue samples were carried on pathological examination. All the tumor tissue contained small cell carcinoma components, there were four cases coexisting with other histologic types of bladder cancers, and 2 of the 9 cases have three different cell components. All patients had muscle-invasive disease at presentation, 4 cases showed lymph nodes metastasis, 3 cases showed invasion of the surrounding seminal vesicle or uterus, and 1 case greatly suspected the liver metastasis. Specimens of all cases underwent immunohistochemistry examination showed that NSE, PCK, Syn, and CD56 were all positive, but LCA was negative. After operation, 3 patients underwent chemotherapy and only one patient received post operational radiotherapy. Patients were followed up range between 3 to 84 months and the median survival time was 33 months. The main reasons of the patients die are tumor recurrence and metastasis, but two patients are still alive. Conclusions SCCB is different from transitional cell carcinoma (TCC) of the bladder. It has its unique cytology, immunohistochemistry and ultrastructural features. Diagnosis depends on pathological examination and immunohistochemistry. Surgery followed by chemotherapy is the main treatment method currently. In view of the disease is easily early metastasis, the overall prognosis for this cancer is poor. Further research is required to understand the molecular pathogenesis so that novel targeted therapies can be developed for this rare cancer.

AB096. Taurine supplementation improves erectile function in rats with streptozotocin-induced type 1 diabetes via amelioration of penile fibrosis and endothelial dysfunction

Objective For patients with diabetes, erectile dysfunction (ED) is common and greatly affects quality of life. However, these patients often exhibit a poor response to first-line oral phosphodiesterase type 5 inhibitors. The aim of this study was to investigate whether taurine, a sulfur-containing amino acid, affects diabetic ED (DED). Methods Type 1 diabetes mellitus was induced in male rats using streptozotocin. After 12 weeks, an apomorphine test was conducted to confirm DED. Only rats with DED were administered taurine or vehicle for four weeks. Age-matched nondiabetic rats were administered saline intraperitoneally for four weeks. Erectile function was evaluated by electrical stimulation of the cavernous nerve. Histologic and molecular alterations of the corpus cavernosum also were analyzed. Results Erectile function was significantly reduced in the diabetic rats compared with in the nondiabetic rats, and was ameliorated in the diabetic rats treated with taurine. The corpus cavernosum of the rats with DED exhibited severe fibrosis and decreased smooth muscle content. Deposition of extracellular matrix proteins was increased in the diabetic rats, while expression of endothelial nitric oxide synthase/cyclic guanosine monophosphate/nitric oxide pathway–related proteins was reduced. Taurine supplementation restored erectile response as well as histologic and molecular alterations. Conclusions Taurine supplementation improves erectile function in rats with DED probably by potential antifibrotic activity. This finding provides evidence for a potential new therapy for DED.

AB156. Taurine ameliorates erectile function in streptozocin-induced type 1 diabetic rats via multiple signaling pathways

Objective PDE5 inhibitors represent the first line therapy for treatment of ED. However, diabetic patients have poorer response compared with normal patients. The aim of this study was to determine whether taurine, a sulfur-containing amino acid, affects diabetic erectile dysfunction. Methods Type 1 diabetes mellitus was induced in male rats by streptozotocin (60 mg/kg, intraperitoneally). After 12 weeks, apomorphine test was conducted to confirm diabetic erectile dysfunction (DED). Only DED rats were administered taurine (400 mg/kg/day, intraperitoneally) or vehicle, for 4 weeks. Age-matched control, nondiabetic, rats were treated with saline intraperitoneally for 4 weeks. At week 16, after a 2-day washout, erectile function was evaluated. Peniles were harvested for Western blot analysis of RhoA, ROCK-1, ROCK-2, eNOS, nNOS, NADPH oxidase subunit gp91phox. Results Erectile function was significantly destroyed in diabetic rats compared with nondiabetic rats and was ameliorated in diabetic rats treated with taurine. In diabetic rats, RhoA, ROCK-1, ROCK-2 and gp91phox protein expressions were increased, whereas eNOS and nNOS expressions were decreased compared with nondiabetic rats, and both were reversed in diabetic rats treated with taurine. Conclusions Taurine improves erectile function in diabetic rats probably by inhibiting RhoA/Rock and activating NOS/NO signaling pathways. This may provide a potential new therapy for diabetic ED.

AB152. Analysis of altered microRNA expression profile in calcium oxalate stone formation

Objective Calcium oxalate stones account for over 80% of urinary stones, while the molecular mechanism of its formation has been unclear. Hyperoxaluria plays an important role in the pathophysiological process of stone formation. The difference of miRNA expression profiles between experimental hyperoxaluric rats and normal rats is analyzed in our study, in order to find out the target genes and signaling pathways in the pathogenesis procedure of hyperoxaluria. Methods Feeding ethylene glycol and ammonium chloride to culture male experimental hyperoxaluric rats as the experimental group, and age-matched male rats were selected as the control group. The oxalate concentration of 24 hour urine of each experimental rat was measured, of which the three highest were selected for microarray test. MicroRNA microarray was applied to evaluate the expression difference between the two groups and screen miRNA whose fold change were above 2.0 or below 0.5. Quantitative real-time PCR (qRT-PCR) technology was used to validate the microarray results. Target prediction, Gene Ontology (GO) analysis and pathway analysis were applied to predict the potential roles of microRNAs in biological processes. Results There are 28 miRNAs differentially expressed with a more than 2.0-fold change. Among these miRNAs, 20 were up-regulated while 8 were down-regulated. After GO analysis and pathway analysis, the insulin resistance pathway and PI3K-Akt signaling pathway were associated with miRNA regulation. Conclusions This study identified differentially expressed miRNAs in hyperoxaluric rats, providing new insights into the role of miRNA in the formation of calcium oxalate stones.

AB224. SaRNA guided inos upregulation improves erectile function of diabetic rats

Purpose Promoter targeted saRNAs mediate sequence specific up-regulation of gene expression. We explored the therapeutic effect of RNA activation mediated iNOS gene activation on improving erectile function in a rat model of diabetes mellitus. Materials and methods An optimal saRNA sequence specific for iNOS promoter was cloned into an adenoviral vector, resulting in AdU6/shiNOS and AdU6/shControl. The corresponding viruses were used to transduce cultured rat cavernous smooth muscle cells. Streptozotocin induced diabetes models were established in rats and used to test the effects of intracavernous delivery of iNOS saRNA viruses on erectile function. iNOS expression in the cavernous smooth muscle cells or penile tissue of treated rats was assessed by reverse transcriptase-polymerase chain reaction and Western blot. Cyclic guanosine monophosphate was analyzed by enzyme-linked immunosorbent assay. Intracavernous pressure in response to cavernous nerve stimulation was measured using a data acquisition system on post-injection days 1, 3, 5, 7, 10 and 14. Results Adenovirus mediated expression of iNOS saRNA caused sustained up-regulation of iNOS in cavernous smooth muscle cells. Intracavernous injection of AdU6/shiNOS activated iNOS expression in vivo and significantly increased peak intracavernous pressure in streptozotocin induced diabetic rats via nitric oxide/intracellular cyclic guanosine monophosphate activation. Conclusions Results show that saRNA mediated iNOS over expression in the penis can restore erectile function in streptozocin diabetic rats via the nitric oxide-cyclic guanosine monophosphate pathway.