Segmental ureterectomy can be performed safely in patients with urothelial carcinoma of distal ureter.

INTRODUCTION: We aimed to compare oncological outcomes by surgery type (segmental ureterectomy [SU] vs. radical nephroureterectomy [RNU]) in a large cohort of patients with upper tract urothelial carcinoma (UTUC) of the distal ureter. METHODS: We performed a retrospective analysis of 219 patients with UTUC of the distal ureter among 931 patients with UTUC who underwent SU and RNU. Clinicopathological outcomes were evaluated. Cancer-specific survival (CSS), overall survival (OS), local recurrence-free survival (RFS), intravesical recurrence-free survival (IVRFS), contralateral recurrence-free survival, and distal metastasis-free survival were assessed by the Kaplan-Meier method and Cox regression, estimating hazard ratios (HR) and 95% confidence intervals (CIs). RESULTS: A total of 179 (81.7%) patients underwent RNU and 40 (18.3%) underwent SU: 85 males (47.5%) with RNU and 17 (42.5%) with SU (p=0.568). The median age with RNU and SU was 71 years (range 31-86) and 70 years (range 46-90), respectively (p=0.499). The T stage of the two groups did not differ (p=0.122), nor did mean tumour length (3.35±2.62 vs. 3.25±2.14; p=0.953), grade (p=0.075), tumour necrosis (p=0.634), or followup time (months) (58.1±8.1 vs. 63.7±3.4; p=0.462). The two groups did not differ in CSS (p=0.358) or OS (p=0.206), and surgery type did not predict CSS (HR 0.862; 95% CI 0.469-1.585; p=0.633) or OS (HR 0.764; 95% CI 0.419-1.392; p=0.379). Local RFS was higher with RNU than SU (96.2% vs. 86.0%; p=0.02), but the groups did not differ in IVRFS (p=0.661), contralateral RFS (p=0.183), or distant metastasis-free survival (p=0.078). On multivariate analysis, SU was associated with local RFS (HR 5.069; 95% CI 1.029-24.968; p=0.046) and distant metastasis-free survival (HR 6.497; 95% CI 1.196-35.283; p=0.03). Local RFS was lower with SU than RNU for patients with pT3-4 stage (p=0.006). CONCLUSIONS: Long-term oncological outcomes were equivalent with SU and RNU in patients with UTUC of the distal ureter. SU affected local recurrence survival, especially with advanced tumour stage, and distant metastasis survival.

The significance of the initial symptom in Chinese patients with upper tract urothelial carcinoma: Regular health examination is still underutilized.

This paper is to elucidate the correlation between different symptoms of UTUC and the clinicopathological characteristics and prognosis. The clinicopathological data of 700 consecutive patients with UTUC who were treated with radical nephroureterectomy were reviewed, and symptoms were categorized into three groups: S1-no direct symptoms, S2-local symptoms (including hematuria and flank pain) and S3-systemic symptoms. We found that the distributions of patients in the S1, S2 and S3 groups were 96 (13.7%), 601 (85.9%) and 3 (0.4%), respectively, and most patients in S1 were incidentally found to have abnormal findings on ultrasound during regular health examination. Altogether, 534 patients (76.3%) presented with gross hematuria, and 111 (15.9%) presented with flank pain. Patients in S1 had a higher rate of hydronephrosis (p < 0.001), ureteral tumors (p < 0.001), worse pre-operative renal function (p = 0.020) and lower tumor stage (p = 0.038). The presence of hematuria was significantly related with renal pelvic tumors (p < 0.001), higher pre-operative eGFR (p = 0.047), papillary tumor architecture (p = 0.005) and less hydronephrosis (p < 0.001); and the presentation of flank pain was correlated with older age (p = 0.008), ureteral location (p < 0.001), hydronephrosis (p < 0.001), sessile architecture (p < 0.001) and higher tumor grade (p = 0.003). The presence of hematuria or flank pain also failed to reach significance as an independent prognostic factor. In conclusion, asymptomatic UTUC patients are featured for more hydronephrosis and lower tumor stage, while patients who presented with flank pain had a higher risk of sessile architecture and higher tumor grade. Regular health examinations might play a useful role in the early detection of UTUC patients with no symptoms.

Detection of urothelial carcinoma, upper tract urothelial carcinoma, bladder carcinoma, and urothelial carcinoma with gross hematuria using selected urine-DNA methylation biomarkers: A prospective, single-center study.

INTRODUCTION: Hematuria is the most common symptom of urothelial carcinomas (UC) but is often idiopathic. Cystoscopy is expensive which involves considerable patient discomfort, and conventional urine cytology for noninvasive UC detection and disease monitoring suffers from poor sensitivity. We aim to evaluate the performance of genes selected from a previous study in detecting UC, especially among patients with gross hematuria, as well as upper tract urothelial carcinoma (UTUC) and bladder carcinoma separately, in voided urine samples. METHODS: Using methylation-specific polymerase chain reaction, we examined the promoter methylation status of 10 genes in voided urine samples among 473 patients at our institution, including 217 UC patients and 256 control subjects. RESULTS: The final combination of VIM, CDH1, SALL3, TMEFF2, RASSF1A, BRCA1, GDF15, and ABCC6 identified UC with a sensitivity of 0.83 and a specificity of 0.60. Additionally, a panel of selected genes (CDH1, HSPA2, RASSF1A, TMEFF2, VIM, and GDF15) identified UTUC with a sensitivity of 0.82 and a specificity of 0.68, while a panel of selected genes (VIM, RASSF1A, GDF15, and TMEFF2) identified bladder carcinoma with a sensitivity of 0.82 and a specificity of 0.53. Remarkably, a different panel (CDH1, SALL3, THBS1, TMEFF2, VIM, and GDF15) identified UC in patients with gross hematuria with 0.89 sensitivity and 0.74 specificity, and sensitivity (0.91) and specificity (0.92) could be achieved when cytology was included. CONCLUSIONS: The selected urine-DNA methylation biomarkers are reliable, noninvasive, and cost-effective diagnostic tools for bladder carcinoma and UTUC, especially among patients with gross hematuria.

Prognostic significance of HALP (hemoglobin, albumin, lymphocyte and platelet) in patients with bladder cancer after radical cystectomy.

The outcome of bladder cancer after radical cystectomy is heterogeneous. We aim to evaluate the prognostic value of HALP (hemoglobin, albumin, lymphocyte and platelet) and explore novel prognostic indexes for patients with bladder cancer after radical cystectomy. In this retrospective study, 516 patients with bladder cancer after radical cystectomy were included. The median follow-up was 37 months (2 to 99 mo). Risk factors of decreased overall survival were older age, high TNM stage, high American Society of Anesthesiologists (ASA) grade and low HALP score. The predictive accuracy was better with HALP-based nomogram than TNM stage (C- index 0.76 ± 0.039 vs. 0.708 ± 0.041). By combining ASA grade and HALP, we created a novel index-HALPA score and found it an independent risk factor for decreased survival (HALPA score = 1, HR 1.624, 95% CI 1.139-2.314, P = 0.007; HALPA score = 2, HR 3.471, 95% CI: 1.861-6.472, P < 0.001).The present study identified the prognostic value of HALP and provided a novel index HALPA score for bladder cancer after radical cystectomy.

Preoperative Prognostic Nutritional Index is a Significant Predictor of Survival with Bladder Cancer after Radical Cystectomy: a retrospective study.

BACKGROUND: To explore the prognostic significance of preoperative prognostic nutritional index (PNI) in bladder cancer after radical cystectomy and compare the prognostic ability of inflammation-based indices. METHODS: We retrospectively analyzed data for 516 patients with bladder cancer who underwent radical cystectomy in our institution between 2006 to 2012. Clinicopathologic characteristics and inflammation-based indices (PNI, neutrophil/lymphocyte ratio [NLR], platelet/lymphocyte ratio [PLR], lymphocyte/monocyte ratio [LMR]) were evaluated by pre-treatment measurements. Overall survival (OS) and progression-free survival (PFS) were estimated by the Kaplan-Meier method and compared by log-rank test. Multivariate analysis with a Cox proportional hazards model was used to confirm predictors identified on univariate analysis. The association between clinicopathological characteristics and PNI or NLR was tested. RESULTS: Among the 516 patients, the median follow-up was 37 months (interquartile range 20 to 56). On multivariate analysis, PNI and NLR independently predicted OS (PNI: hazard ratio [HR] = 1.668, 95% CI: 1.147-2.425, P = 0.007; NLR: HR = 1.416, 95% CI:1.094-2.016, P = 0.0149) and PFS (PNI: HR = 1.680, 95% CI:1.092-2.005, P = 0.015; NLR: HR = 1.550, 95% CI:1.140-2.388, P = 0.008). Low PNI predicted worse OS for all pathological stages and PFS for T1 and T2 stages. Low PNI was associated with older age (>65 years), muscle-invasive bladder cancer, high American Society of Anesthesiologists grade and anemia. CONCLUSION: PNI and NLR were independent predictors of OS and PFS for patients with bladder cancer after radical cystectomy and PNI might be a novel reliable biomarker for bladder cancer.

Higher programmed cell death 1 ligand 1 (PD-L1) mRNA level in clear cell renal cell carcinomas is associated with a favorable outcome due to the active immune responses in tumor tissues.

Renal cell carcinoma is one of the most common urological tumors. The role of programmed cell death 1 ligand 1 (PD-L1) in renal cell carcinomas in predicting outcome of the patients is yet unclear. We analyzed the clinical and RNA-seq data of 522 kidney clear cell cancer, 259 kidney papillary cell carcinoma and 66 kidney chromophobe patients from The Cancer Genome Atlas (TCGA) database. In kidney clear cell cancer patients with high PD-L1 mRNA level and low PD-L1 mRNA level in tumors, the median overall survival periods were 45.0 and 37.1 months respectively (p=0.002). Multivariate Cox regression tests found that PD-L1 mRNA level in tumor was an independent predictor for overall survival status in kidney clear cell cancer patients (HR=0.7, 95% CI 0.5-0.9, p=0.007). However, no significant difference in overall survival status was found between high and low PD-L1 groups in kidney papillary cell carcinoma and kidney chromophobe cohorts. Gene-set enrichment analysis on the data from databases of TCGA and GSE53757 dataset in Gene Expression Omnibus databases showed that several pathways relating to immunological functions were activated in kidney clear cell cancers with high PD-L1 mRNA expression, and glycolysis and epithelial-mesenchymal transition pathways relating to tumor progression and metastasis were increased in kidney clear cell cancers with low PD-L1 mRNA level. In conclusion, higher PD-L1 mRNA level in kidney clear cell cancer tissues was associated with a favorable outcome due to the higher immunological responses in tumor tissues.

[Establishment of Human Chronic Lymphocytic Leukemia Model in the BALB/c Nude Mice by Using MEC-1 and HG3 Cell Lines].

OBJECTIVE: To explore the the optimal condition for establishing immune deficiency mouse(BALB/c) model with CLL via subcutaneous inoculation of human chronic lymphocytic leukemia (CLL) cells at different inoculative locations and different cell concentrations. METHODS: Firstly, Two CLL cell lines (MEC-1-GFP and HG3-GFP)with the green fluorescent protein (GFP) were established by lentivirus system respectively, and then the MEC-1-GFP cells (5×107/ml) were inoculated into forelimb, hindlimb and abdomen to observe the tumorigenesis. Secondly, the MEC-1-GFP and HG3-GFP cells with same density (5×107/ml) were inoculated into forelimb to compare the time and rate of tumor formation. Thirdly, the MEC-1-GFP cells (1×107/ml) and HG3-GFP cells (5×107/ml) were inoculated into forelimb to compare the time and rate of tumor formation at different inoculative density. After observation for 5 weeks, the peripheral blood was collected and treated with EDTA and erythrocytolysin, then the of GFP positive cells were detected by flow cytomety. Meanwhile, the tumor-bearing mice were killed, and the tumors were isolated and cut into slices for histopathological examination. RESULTS: MEC-1-GFP and HG3-GFP cell lines were successfully established, and after inocutation of MEC-1-GFP cells with 5×107/ml the xenograt tumors were formated in forelimb, hindlimb and abdomen of mice, especially in the forelimb with a higher tumorigenic rate. In addition, the inoculation of same density of MEC-1-GFP and HG3-GFP cells (5×107/ml) also resulted in xenograft in forelimb, and the tumorigenic rate reached to 80% after 5 weeks. Moreover, the inocutation of MEC-1-GFP and HG3-GFP cells with 1×107 and 5×107/ml respectively also effectively resulted to xenograft tumor in forelimb. The flow cytometry showed that there was no MEC-1-GFP and HG3-GFP cells in peripheral blood, while histopathological examination demonstrated CLL cell metastasis towards peritoneal cavity. CONCLUSION: The BALB/c nude mouse model is successfully established by subcutaneous injection of MEC-1-GFP and HG3-GFP cells. This model is a useful tool to explore the pathogenic mechanism.

Association between FBP1 and hypoxia-related gene expression in clear cell renal cell carcinoma.

Fructose-1,6-bisphosphatase 1 (FBP1) is a rate-limiting enzyme in gluconeogenesis. Recently, the catalytic activity-independent function of FBP1, hypoxia-induced factor (HIF) repression in the nucleus, was identified. The aim of the present study was to investigate the association between FBP1 and hypoxia-related gene expression in clear cell renal cell carcinoma (ccRCC). The protein expression levels of FBP1, HIF-1α, HIF-2α, erythropoietin (EPO) and carbonic anhydrase IX (CA9) were assessed by immunohistochemical staining of ccRCC paraffin blocks from 123 patients using the tissue microarray technique. The expression level of FBP1 was then correlated with various clinicopathological factors, and the protein expression levels of HIF-1α, HIF-2α, EPO and CA9. Clinicopathological factors, including age, gender, T stage and Fuhrman grade, were not significantly different between patients with low and high FBP1 expression in ccRCC (P>0.05). FBP1 protein expression level was significantly correlated with the expression levels of HIF-1α (P=0.005) and EPO (P=0.010), but not significantly correlated with the expression levels of HIF-2α (P=0.123) and CA9 (P=0.513) in ccRCC tissues. The current findings confirm the association between FBP1 and hypoxia-related gene expression, and may facilitate understanding of the mechanisms of ccRCC tumorigenesis.

UBE2T silencing suppresses proliferation and induces cell cycle arrest and apoptosis in bladder cancer cells.

Ubiquitin-conjugating enzyme E2T (UBE2T), a member of the ubiquitin-conjugating E2 family in the ubiquitin-proteasome pathway, has been reported to be overexpressed in certain tumor types and to have an important role in the Fanconi anemia pathway. In the present study, the expression of UBE2T and its association with bladder cancer were investigated; to the best of our knowledge, this has not been reported previously. Immunohistochemistry and western blot analysis demonstrated that UBE2T was significantly upregulated in bladder cancer tissues and cell lines compared with adjacent normal bladder tissues and a normal human urinary tract epithelial cell line, respectively. UBE2T was detectable in the nuclei and cytoplasm of cancer cells, exhibiting stronger expression in the nuclei. A UBE2T-siRNA-expressing lentivirus was constructed and used to infect human bladder cancer 5637 cells, in order to examine the role of UBE2T in bladder cancer cell growth in vitro. The knockdown of UBE2T significantly decreased bladder cancer cell proliferation and colony formation. Furthermore, UBE2T silencing induced cell cycle arrest at G2/M phase and increased cell apoptosis. Therefore, UBE2T serves an important role in the growth of bladder cancer cells, and may be considered as a potential biomarker and therapeutic target for bladder cancer.

Crystal structure of 1,7,8,9-tetra-chloro-4-(3,5-di-chloro-benz-yl)-10,10-dimeth-oxy-4-aza-tri-cyclo-[5.2.1.0(2,6)]dec-8-ene-3,5-dione.

In the title compound, C17H11Cl6NO4, the configuration of the cyclo-alkene skeleton is endo,cis. The benzene ring is twisted by 58.94 (8)° from the attached pyrrolidine ring. Two carbonyl groups play a key role in the crystal packing. A short inter-molecular C⋯O distance of 3.017 (3) Šreveals that one carbonyl group is involved in dipole-dipole inter-actions, which link two adjacent enanti-omers into an inversion dimer. Another carbonyl group provides an acceptor for the weak inter-molecular C-H⋯O hydrogen bonds which link these dimers into layers parallel to (011).

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.

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.

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.

Tissue engineering penoplasty with biodegradable scaffold Maxpol-T cografted autologous fibroblasts for small penis syndrome.

In this study, we investigated the safety and efficacy of a poly acid-co-glycolide biodegradable scaffold (Maxpol-T) coated by autologous fibroblasts (AF) for penile girth enlargement in small penis syndrome (SPS). Eighty patients with SPS were enrolled in a clinical study at 2 medical centers; 69 patients completed the study protocol. Scrotal skin was harvested under local anesthesia, and AFs were cultured and seeded on a Maxpol-T scaffold; the cografted scaffold was implanted under the Buck's fascia of penile shaft via a circumcising incision. Patients were followed up at 1, 3, and 6 months to evaluate penile girth changes. Patient satisfaction was assessed via Visual Analogue Scale and scored on the International Index of Erectile Function-5 (IIEF-5). Mean preoperative penile girth in the flaccid and erect state was 8.18 ± 0.83 cm and 10.26 ± 1.22 cm, respectively. At the 6-month postoperative follow-up, mean penile girth in the flaccid and erect state was increased to 12.19 ± 1.27 cm and 13.18 ± 1.31 cm, respectively (P < .001 for change in both flaccid and erect state). Sixty-five patients (94.2%) reported satisfaction with the procedure. Among them, 4 cases (5.8%) were dissatisfied, 7 cases (10.1%) were satisfied, 26 cases (37.7%) were very satisfied, and 32 cases (46.4%) were extremely satisfied. All men maintained IIEF-5 scores of more than 22. Complications included prolonged subcutaneous edema in 3 patients (4.3%) and pinpoint erosion at the suture area in 3 patients (4.3%). Implantation of autologous fibroblasts seeded on a Maxpol-T collagen scaffold holds promise as a safe and novel technique for penile girth enhancement in patients with SPS.

Bioactive Nortriterpenoids from Schisandra grandiflora.

Three new nortriterpenoids, schigrandilactones A-C (1-3), along with eight known compounds, were isolated from an organic solvent extract of Schisandra grandiflora. Compounds 1 and 2 feature a spirocyclic moiety in their structures, and compound 3 was characterized with a new oxygenated pattern. The relative configurations of 1 and 3 were determined through single-crystal X-ray experiments. In addition, compounds 1 and 2 displayed cytotoxic activity against two human cancer cell lines, and compound 3 showed anti-HIV-1 inhibition in infected C8166 cells.

Daphniphyllum and diterpenoid alkaloids from Daphniphyllum longeracemosum.

Four new Daphniphyllum alkaloids, daphlongamines A-D (1-4), were isolated from the fruits of Daphniphyllum longeracemosum. Their structures were characterized by spectroscopic methods, especially 2D NMR techniques. A single-crystal X-ray diffraction analysis was used to confirm the stereochemistry of 3. Remarkably, this is the first report of aconitine- and veatchine-type diterpenoid alkaloids from the genus Daphniphyllum.

[Studies on chemical constituents in stem of Dendrobium chrysotoxum].

OBJECTIVE: To investigate the chemical constituents of Dendrobium chrysotoxum. METHOD: The chemical constituents were isolated by various column chromatographic methods and structurally elucidated by spectral evidences. RESULT: Ten compounds were obtained and identified as (+)-syringare sinol (1), 5alpha, 8alpha-epidioxy-24( R)-methycholesta-6, 22-dien-3beta-ol (2), trans-3-(4-hydroxy-3-methoxyphenyl)-acrylic acid octacosyl ester (3), defusin (4), 3, 4-dihydroxy benzoic acid (5), 3, 4-dimethoxy-benzoic acid (6), vanillic acid (7), 3, 4-dimethoxy-benzoic acid methyl ester (8), 3, 5-dibromo-2-aminobenzaldehyde (9), heptadecanoic acid 2, 3-dihydroxy-propyl ester (10). CONCLUSION: Compounds 1, 2 and 6-10 were isolated from this plant for the first time.