Hyperforin regulates renal fibrosis via targeting the PI3K-AKT/ICAM1 axis.

OBJECTIVE: To explore the role and mechanism of hyperforin (one of the active components of Sophora flavescens) in renal fibrosis. METHODS: The active compounds and target proteins of Sophora flavescens were first screened through TCMSP (https://tcmsp-e.com/). The renal fibrosis-related genes were analyzed through GeneCards (https://www.genecards.org/). The differentially expressed genes (DEGs) in renal fibrosis in GEO dataset GSE156181 were obtained. Metascape was applied for target protein enrichment analysis. TGF-ß1-stimulated renal tubular epithelial cells were used for renal fibrosis cell model establishment. The unilateral ureteral obstruction (UUO) mouse model was used for the renal fibrosis in vivo model. Cell viability was detected using an MTT assay. Immunofluorescence staining was employed to detect cell morphology changes and the expression of α-SMA and collagen I. Hematoxylin and eosin (H&E) and Masson staining were employed to determine the renal morphologic change. qRT-PCR or Western blotting was applied to determine the expression levels of the target proteins. RESULTS: After intersecting the analysis results of TCMSP, GeneCards, and dataset GSE156181, hyperforin targeting ICAM1 was identified. Metascape pathway enrichment analysis results revealed that the effective compounds of Sophora flavescens were tightly associated with extracellular matrix (ECM) remodeling and inflammatory response. MTT assay demonstrated that hyperforin had no toxic effect on cells. Immunofluorescence staining results evidenced that hyperforin could partially restore TGF-ß1-induced epithelial-mesenchymal transition (EMT), the PI3K/AKT pathway activation, and ICAM1 upregulation, and these effects of hyperforin could be reversed by ICAM1 overexpression. While the PI3K/AKT pathway activator IGF-1 effectively reversed the EMT inhibition effect of hyperforin on renal tubular epithelial cells. Moreover, the UUO mouse model further confirmed that hyperforin reduced renal fibrosis. CONCLUSION: Hyperforin inhibited renal fibrosis via the PI3K/AKT/ICAM1 axis.

Factors associated with anxiety and depression in patients with erectile dysfunction: a cross-sectional study.

BACKGROUND: Few studies have investigated factors associated with anxiety and depression among patients with erectile dysfunction (ED). This study aimed to investigate associated factors and the prevalence of anxiety and depression in this special group in China. METHODS: Data from 511 patients with ED aged 18-60 years were collected between July 2021 and April 2022. The 5-item International Index of Erectile Function (IIEF-5) questionnaire, self-rating anxiety scale (SAS) and self-rating depression scale (SDS) were used to evaluate erectile function, anxiety and depression, respectively. Univariate analysis and multivariate linear regression analyses were used to explore the associated factors of depression and anxiety. RESULTS: The prevalence of anxiety and depression among ED patients was 38.16% and 64.97%, respectively. The mean anxiety index score was 47.37 ± 6.69 points, and the mean depression index was 54.72 ± 9.10 points. Multiple linear regression analysis showed that worse ED, low education level, and smoking were positively associated with increased risk of anxiety and depression. In addition, younger age, longer onset time, and irregular sleep were positively associated with high risk of anxiety, and irregular exercise was associated with severe depression. CONCLUSIONS: The prevalence of depression and anxiety in ED patients is high, and the severity of ED, age, education level, smoking, onset time, regular sleep, and exercise were associated with anxiety or depression. Reversible risk factors should be avoided and individualized psychological support services are necessary for ED patients.

Artificial Neural Network-Based Activities Classification, Gait Phase Estimation, and Prediction.

Gait patterns are critical to health monitoring, gait impairment assessment, and wearable device control. Unrhythmic gait pattern detection under community-based conditions is a new frontier in this area. The present paper describes a high-accuracy gait phase estimation and prediction algorithm built on a two-stage artificial neural network. This work targets to develop an algorithm that can estimate and predict the gait cycle in real time using a portable controller with only two IMU sensors (one on each thigh) in the community setting. Our algorithm can detect the gait phase in unrhythmic conditions during walking, stair ascending, and stair descending, and classify these activities with standing. Moreover, our algorithm is able to predict both future intra- and inter-stride gait phases, offering a potential means to improve wearable device controller performance. The proposed data-driven algorithm is based on a dataset consisting of 5 able-bodied subjects and validated on 3 different able-bodied subjects. Under unrhythmic activity situations, validation shows that the algorithm can accurately identify multiple activities with 99.55% accuracy, and estimate ([Formula: see text]: 6.3%) and predict 200-ms-ahead ([Formula: see text]: 8.6%) the gait phase percentage in real time, which are on average 57.7 and 54.0% smaller than the error from the event-based method in the same conditions. This study showcases a solution to estimate and predict gait status for multiple unrhythmic activities, which may be deployed to controllers for wearable robots or health monitoring devices.

What Influences Coital Frequency Among Chinese Men?: A Cross-Sectional Study.

INTRODUCTION: There are many Western reports on factors influencing coital frequency among men. However, no articles could be found about the factors influencing sexual activity among Chinese men. AIM: The aim of this study was to identify the factors that influence the coital frequency of Chinese men. MAIN OUTCOME MEASURES: The main outcome measures included self-reported monthly coital frequency, age, occupation, education level, andrology-related scales and dietary habits. METHODS: Data for 1,407 men aged 18-79 years were collected in the Health Management Center of the Third Xiangya Hospital of Central South University from January 2019 to May 2019. The respondents completed the questionnaires independently or with the help of an interviewer (who read or explained the questionnaires to them) to analyse the factors that influence coital frequency. RESULTS: In the previous 6 months, the sample had a mean monthly coital frequency (±SD) of 4.34 ± 3.18. Univariate logistic regression results indicated that the number of children (P = 0.004), IIEF-5 scores (P <0.001), EHSs (P <0.001) and frequency of milk consumption (P = 0.001) were associated with more frequent sexual activity. These statistical associations did not change after further adjustment for age, occupation, and reproductive history. We observed that the frequency of sexual activity showed an increasing trend with a greater number of children, higher IIEF-5 scores, higher EHSs and greater frequency of milk consumption (test for trend, P<0.05). Both univariate and multivariate analysis results indicated that the frequency of sexual activity decreased with increasing age (test for trend, P<0.001). CONCLUSION: The coital frequency of Chinese men is associated with erectile function, anthropometric parameters, age, occupation, and dietary habits. Xiang Y, Peng J, Yang J, et al. What Influences Coital Frequency Among Chinese Men?: A Cross-Sectional Study. Sex Med 2021;9:100363.

The m6A methyltransferase METTL3 regulates autophagy and sensitivity to cisplatin by targeting ATG5 in seminoma.

BACKGROUND: Our previous work shows Autophagy enhanced resistance to cisplatin in seminoma. The expression of the N6-methyladenosine (m6A) methyltransferases METTL3 was significantly increased in the cisplatin-resistant TCam-2 cell line of seminoma. We aimed to investigate the role of m6A methylation in autophagy and the chemosensitivity of seminoma cells. METHODS: Plasmid and siRNA were used to overexpress and knockdown METTL3. Autophagy was detected by western blot and immunofluorescence, respectively. The expression of downstream targets of METTL3 was detected by quantitative real-time PCR (qRT-PCR) and western blot, and the m6A level of them was detected by MeRIP-qPCR. Chemosensitivity of the TCam-2 cell line was identified through MTT assay. RESULTS: Upon METTL3 overexpression, autophagy of TCam-2 cell line was enhanced and its sensitivity to cisplatin was decreased. The use of autophagy inhibitors 3-methyladenine (3-MA) could reverse the protective effect of METTL3 on TCam-2 cells. We found that the up-regulation of METTL3 could increase the m6A modification level of ATG5 transcript, thus increased expression of ATG5. Moreover, knockdown of ATG5 reduced METTL3-induced autophagy, suggesting that ATG5 was a potential target for METTL3 to promote autophagy. CONCLUSIONS: In summary, our research unveiled the unique mechanism by which m6A methylation regulates autophagy and chemosensitivity of the TCam-2 cell line and METTL3 was a potential target to overcome the cisplatin resistance of seminoma.

METTL3 potentiates resistance to cisplatin through m6 A modification of TFAP2C in seminoma.

Testicular germ cell tumours (TGCTs) rank as the most common malignancy in men aged 20-34 years, and seminomas are the most type of TGCTs. As a crucial anti-tumour agent with explicit toxicity, cisplatin may render resistance through intertwined mechanisms, even in disease entities with high curative ratio, such as seminoma. Previously, we established cisplatin-resistant seminoma TCam-2 (TCam-2/CDDP) cells and showed that epigenetic regulations, such as non-coding RNA (ncRNA) interactions, might orchestrate cell fate decisions in the cisplatin treatment context in seminoma. N6-methyladenosine (m6A) is the most prevalent internal modification in mRNA. In the present study, we assessed cisplatin resistance in seminoma from the perspective of m6 A, another manner of epigenetic modification. The global m6 A enrichment of TCam-2 and TCam-2/CDDP was depicted. Then, we elucidated whether transcription factor-activating enhancer-binding protein 2C (TFAP2C) was functionally m6 A-modified by methyltransferase-like protein 3 (METTL3), which acted as an m6 A 'writer', and insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1), which acted as an m6 A 'reader'. Enhanced stability of TFAP2C mRNA promoted seminoma cell survival under cisplatin treatment burden probably through up-regulation of DNA repair-related genes. Hopefully, this study will help improve our understanding of the subtleties of the tumour cellular coping strategy in response to chemotherapy. Targeting factors that are involved in m6 A methylation may be an effective strategy for circumventing cisplatin resistance in seminoma.

Effects of Inorganic Arsenic on Human Prostate Stem-Progenitor Cell Transformation, Autophagic Flux Blockade, and NRF2 Pathway Activation.

BACKGROUND: Inorganic arsenic (iAs) is an environmental toxicant associated with an increased risk of prostate cancer in chronically exposed populations worldwide. However, the biological mechanisms underlying iAs-induced prostate carcinogenesis remain unclear. OBJECTIVES: We studied how iAs affects normal human prostate stem-progenitor cells (PrSPCs) and drives transformation and interrogated the molecular mechanisms involved. METHODS: PrSPCs were enriched by spheroid culture from normal human primary or immortalized prostate epithelial cells, and their differentiation capability was evaluated by organoid culture. Microarray analysis was conducted to identify iAs-dysregulated genes, and lentiviral infection was used for stable manipulation of identified genes. Soft agar colony growth assays were applied to examine iAs-induced transformation. For in vivo study, PrSPCs mixed with rat urogenital sinus mesenchyme were grafted under the renal capsule of nude mice to generate prostatelike tissues, and mice were exposed to 5 ppm (∼65µM) iAs in drinking water for 3 months. RESULTS: Low-dose iAs (1µM) disturbed PrSPC homeostasis in vitro, leading to increased self-renewal and suppressed differentiation. Transcriptomic analysis indicated that iAs activated oncogenic pathways in PrSPCs, including the KEAP1-NRF2 pathway. Further, iAs-exposed proliferative progenitor cells exhibited NRF2 pathway activation that was sustained in their progeny cells. Knockdown of NRF2 inhibited spheroid formation by driving PrSPC differentiation, whereas its activation enhanced spheroid growth. Importantly, iAs-induced transformation was suppressed by NRF2 knockdown. Mechanistically, iAs suppressed Vacuolar ATPase subunit VMA5 expression, impairing lysosome acidification and inhibiting autophagic protein degradation including p62, which further activated NRF2. In vivo, chronic iAs exposure activated NRF2 in both epithelial and stroma cells of chimeric human prostate grafts and induced premalignant events. CONCLUSIONS: Low-dose iAs increased self-renewal and decreased differentiation of human PrSPCs by activating the p62-NRF2 axis, resulting in epithelial cell transformation. NRF2 is activated by iAs through specific autophagic flux blockade in progenitor cells, which may have potential therapeutic implications. https://doi.org/10.1289/EHP6471.

Integrative Analysis of MicroRNA and Gene Interactions for Revealing Candidate Signatures in Prostate Cancer.

MicroRNA (miRNA)-gene interactions are well-recognized as involved in the progression of almost all cancer types including prostate cancer, which is one of the most common cancers in men. This study explored the significantly dysregulated genes and miRNAs and elucidated the potential miRNA-gene regulatory network in prostate cancer. Integrative analysis of prostate cancer and normal prostate transcriptomic data in The Cancer Genome Atlas dataset was conducted using both differential expression analysis and weighted correlation network analysis (WGCNA). Thirteen genes (RRM2, ORC6, CDC45, CDKN2A, E2F2, MYBL2, CCNB2, PLK1, FOXM1, CDC25C, PKMYT1, GTSE1, and CDC20) were potentially correlated with prostate cancer based on functional enrichment analyses. MiRNAs targeting these genes were predicted and eight miRNAs were intersections between those miRNAs and the hub miRNAs obtained from miRNA WGCNA analysis. Three genes (E2F2, RRM2, and PKMYT1) and four miRNAs (hsa-mir-17-5p, hsa-mir-20a-5p, hsa-mir-92a-3p, and hsa-mir-93-5p) were key factors according to the interaction network. RRM2 and PKMYT1 were significantly related to survival. These findings partially elucidated the dysregulation of gene expressions in prostate cancer. Efficient manipulations of the miRNA-gene interactions in prostate cancer may be exploited as promising therapeutics.

Quasi-Direct Drive Actuation for a Lightweight Hip Exoskeleton with High Backdrivability and High Bandwidth.

High-performance actuators are crucial to enable mechanical versatility of wearable robots, which are required to be lightweight, highly backdrivable, and with high bandwidth. State-of-the-art actuators, e.g., series elastic actuators (SEAs), have to compromise bandwidth to improve compliance (i.e., backdrivability). We describe the design and human-robot interaction modeling of a portable hip exoskeleton based on our custom quasi-direct drive (QDD) actuation (i.e., a high torque density motor with low ratio gear). We also present a model-based performance benchmark comparison of representative actuators in terms of torque capability, control bandwidth, backdrivability, and force tracking accuracy. This paper aims to corroborate the underlying philosophy of "design for control", namely meticulous robot design can simplify control algorithms while ensuring high performance. Following this idea, we create a lightweight bilateral hip exoskeleton to reduce joint loadings during normal activities, including walking and squatting. Experiments indicate that the exoskeleton is able to produce high nominal torque (17.5 Nm), high backdrivability (0.4 Nm backdrive torque), high bandwidth (62.4 Hz), and high control accuracy (1.09 Nm root mean square tracking error, 5.4% of the desired peak torque). Its controller is versatile to assist walking at different speeds and squatting. This work demonstrates performance improvement compared with state-of-the-art exoskeletons.

Testis developmental related gene 1 regulates the chemosensitivity of seminoma TCam-2 cells to cisplatin via autophagy.

We previously identified testis developmental related gene 1 (TDRG1), a gene implicated in proliferation of TCam-2 seminoma cells. Recent evidence has revealed that autophagy influences the chemosensitivity of cancer cells to chemotherapy. However, whether TDRG1 protein regulates autophagy in seminoma cells and influences their sensitivity to cis-dichlorodiammine platinum (CDDP) remains unknown. In this study, we used TCam-2 cells and male athymic BALB/c nude mice with xenografts of TCam-2 cells to investigate autophagy, cell viability, apoptosis and the p110ß/Rab5/Vps34 (PI3-kinase Class III) pathway under the conditions of TDRG1 overexpression or knockdown and with or without CDDP treatment. We found that TDRG1 upregulation promoted autophagy in both TCam-2 cells and seminoma xenografts via p110ß/Rab5/Vps34 activation. Inhibition of autophagy reduced cell viability and promoted apoptosis during CDDP treatment of TCam-2 cells. Similarly, TDRG1 knockdown inhibited autophagy, reduced cell viability and promoted apoptosis during CDDP treatment of TCam-2 cells. TDRG1 knockdown inhibited tumour growth and promoted apoptosis in TCam-2 cell xenografts, whereas TDRG1 overexpression had the opposite effect. According to these results, we propose that high expression of TDRG1 promotes autophagy through the p110ß/Rab5/Vps34 pathway in TCam-2 cells. TDRG1 overexpression promotes autophagy and leads to CDDP resistance, whereas TDRG1 knockdown inhibits autophagy and promotes chemosensitivity to CDDP both in vivo and in vitro. This study has uncovered a novel role of TDRG1 in reducing chemoresistance during CDDP treatment and provides potential therapeutic strategies for the treatment of human seminoma.

Synthesis of a novel platinum(II) complex with 6,7-dichloro-5,8-quinolinedione and the study of its antitumor mechanism in testicular seminoma.

A new platinum(II) complex, [Pt(ClClQ)(DMSO)Cl] (1), utilizing 6,7-dichloro-5,8-quinolinedione (ClClQ) as a ligand, has been synthesized and fully characterized. Single-crystal X-ray diffraction and other spectroscopic and analytical methods revealed that the coordination geometry of Pt(II) in complex 1 can also be described as a four-coordinated square planar geometry. The aim of the study was to explore the in vitro anticancer properties of complex 1. Our studies showed that complex 1 can regulate the viability of testicular seminoma cells in vitro, including cell proliferation and apoptosis. We further observed negative regulation by complex 1 of the expression levels of the key elements in the phosphoinositide-3 kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3ß (GSK3ß) pathway, including phosphorylated phosphoinositide-3 kinase (p-PI3K), phosphorylated protein kinase B(p-Akt) and phosphorylated glycogen synthase kinase-3ß (p-GSK3ß). Moreover, the negative effect of complex 1 was reversed by LiCl, a GSK3ß-specific inhibitor of the PI3K signaling pathway. Meanwhile, the levels of Bcl2 associated death promoter (Bad), cytochrome c, active-caspase-3 and active-caspase-9 increased significantly. In conclusion, we observed that complex 1 can regulate the viability of testicular seminoma cells through the PI3K/Akt/GSK3ß signaling pathway and the mitochondria-mediated apoptotic pathway in vitro, and thus, complex 1 may have potential for use as a drug in the treatment of testicular germ cell tumors.

Long non-coding RNA H19 promotes TDRG1 expression and cisplatin resistance by sequestering miRNA-106b-5p in seminoma.

The role of TDRG1 in tumorigenesis and the progression of seminoma, as well as its role in regulating chemosensitivity of seminoma to cisplatin through the PI3K/Akt/mTOR signaling pathway, has been previously defined. However, the detailed mechanism underlying TDRG1 expression and concomitant chemoresistance conditions are unknown. Furthermore, it has been reported that non-protein-coding RNAs play an important role in a variety of vital processes including cellular chemosensitivity. However, the role of non-protein-coding RNAs in regulating the chemosensitivity of seminoma remains unknown. In this study, using microarray analysis, we found that long non-coding RNA H19 was upregulated while miRNA-106b-5p was downregulated in an established cisplatin-resistant TCam-2 cell line. Moreover, H19 acts as a miRNA-106b-5p sponge and thus impairs the function of miRNA-106b-5p on its target gene, TDRG1. Based on these findings, we propose that H19 promotes the expression of TDRG1 by sequestering miRNA-106b-5p and uses this mechanism to facilitate cell survival in cisplatin-based chemotherapeutic conditions. These findings elucidate the mechanisms, at least partially, applied to deregulate TDRG1 and cisplatin sensitivity, and may provide new therapeutic possibilities for chemoresistant seminoma.

TDRG1 regulates chemosensitivity of seminoma TCam-2 cells to cisplatin via PI3K/Akt/mTOR signaling pathway and mitochondria-mediated apoptotic pathway.

We previously identified TDRG1 (testis developmental related gene 1), a novel gene with exclusive expression in testis, promoted the proliferation and progression of cultured human seminoma cells through PI3K/Akt/mTOR signaling. As increasing evidence reveal that aberrant activation of this signaling is involved in cisplatin resistance. Then, in this study, we further explored whether TDRG1 regulated the chemosensitivity of seminoma TCam-2 cells to cisplatin. Our researches showed TDRG1 could regulate the viability of TCam-2 cells following cisplatin treatment in vitro through control of both cell apoptosis and cell cycle. Mechanistically, we observed TDRG1 positively regulated the expression levels of the key elements in PI3K/Akt/mTOR pathway including p-PI3K, p-Akt and p-mTOR and also affected the translocation of nuclear p-Akt in TCam-2 cells during cisplatin treatment. Meanwhile, the levels of Bad, cytochrome c, caspase-9 ratio (activated/total), caspase-3 ratio (activated/total) and cleaved-PARP were negatively modulated by TDRG1, which meant the involvement of mitochondria-mediated apoptotic pathway. Furthermore, we found the effect of TDRG1 knockdown or TDRG1 overexpression could be reversed by IGF-1, a PI3K signaling activator, or LY294002, a inhibitor of this pathway, respectively. Similar effects of TDRG1 on cisplatin chemosensitivity and associated molecular mechanism were also confirmed in vivo by employing xenograft assays. In addition, the positive correlation between TDRG1 and p-PI3K, or p-Akt, was found in tumor tissues from seminoma patients. In conclusion, we uncover that TDRG1 regulates chemosensitivity of TCam-2 cells to cisplatin through PI3K/Akt/mTOR signaling and mitochondria-mediated apoptotic pathway both in vitro and in vivo.

TDRG1 functions in testicular seminoma are dependent on the PI3K/Akt/mTOR signaling pathway.

Human testis development-related gene 1 (TDRG1) is a recently identified gene that is expressed exclusively in the testes and promotes the development of testicular germ cell tumors. In this study, the role of TDRG1 in the development of testicular seminoma, which is the most common testicular germ cell tumor, was further investigated. Based on polymerase chain reaction, Western blotting, and immunohistochemistry tests, both gene and protein expression levels of TDRG1 were significantly upregulated in testicular seminoma tissues compared with normal testicular tissues. Additionally, the levels of phosphoinositide-3 kinase (PI3K)/p110 and Akt phosphorylation were dramatically upregulated in testicular seminoma tissues. Accordingly, in our cell experiment, seminoma TCam-2 cells were subjected to different treatments: the TDRG1 knockout, TDRG1 overexpression, PI3K inhibition (LY294002 administration), or PI3K activation (insulin-like growth factor-1 administration). Cell proliferation, the proliferation index, apoptosis rate, cell adhesive capacity, and cell invasion capability were assessed. Cells with both TDRG1 knockout and PI3K inhibition exhibited decreased cell proliferation, proliferation indexes, cell adhesion capacity, and cell invasion capability and increased apoptosis rates. Most of these effects were reversed by TDRG1 overexpression or PI3K activation, indicating that both TDRG1- and PI3K-mediated signaling promote proliferation and invasion of testicular seminoma cells. The knockout of TDRG1 significantly decreased the phosphorylation levels of PI3K/p85, PI3K/p110, Akt, and mammalian target of rapamycin (mTOR; Ser(2448)). Except for PI3K/p110, TDRG1 overexpression had the opposite effects on phosphorylation levels. Phosphorylated mTOR at Ser(2481) and Thr(2446) was not affected by TDRG1 or PI3K in our tests. Thus, these results indicate that TDRG1 promotes the development and migration of seminoma cells via the regulation of the PI3K/Akt/mTOR signaling pathway; this contributes to an understanding of the precise mechanisms underlying the development and migration of seminomas and lays a theoretical foundation for the development of appropriate therapies.

Bias in Evaluating Erectile Function in Lifelong Premature Ejaculation Patients with the International Index of Erectile Function-5.

INTRODUCTION: Lifelong premature ejaculation (LPE) is the most important ejaculating dysfunction. Relatively little is known about erectile function in this population. AIMS: We attempted to evaluate the erectile function of patients with LPE using the International Index of Erectile Function-5 (IIEF-5) to determine if it is sufficiently reliable and accurate to make such an assessment. METHODS: A total of 406 patients with LPE were enrolled in our study. The participants voluntarily answered the Premature Ejaculation Diagnostic Tool (PEDT) and IIEF-5 questionnaires and underwent a full history evaluation and clinical examination by doctors. Their answers were converted into data analyzed by a statistic software. MAIN OUTCOME MEASURES: The patients were diagnosed with LPE based on the diagnostic criteria and PEDT scores. The intravaginal ejaculation latency time was recorded according to patient self-reports. The IIEF-5 was used to evaluate their erectile function. Thorough history and clinical examination helped doctors make more correct diagnoses of erectile dysfunction (ED). RESULTS: Of the 406 patients, 70 (17.24%) patients had ED, as confirmed by doctors. IIEF-5 was accurate for the assessment of the erectile function of LPE patients when the cutoff was decreased to 15.5. Question 5 (1.34 ± 0.53) was the main reason for the drop in the total IIEF-5 score. Questions 1 and 5 shared low consistency with the other three IIEF-5 items, thus they lowered the reliability of the IIEF-5 scores. These questions created a confounding bias that decreased the diagnostic threshold of IIEF-5. However, they could not be removed from the IIEF-5 because they did not reduce its diagnostic accuracy in patients with LPE. CONCLUSIONS: Bias from questions 1 and 5 lowered the reliability of the IIEF-5 scores; however, it did not reduce the diagnostic accuracy of the IIEF-5. The recommendation is to edit questions 1 and 5 when they are applied to populations with LPE.

In vitro study on shRNA-mediated reduction of testis developmental related gene 1 expression and its effects on the proliferation, invasion and apoptosis of NTERA-2 cells.

Testis developmental related gene 1 (TDRG1) is a novel human testis-specific gene. TDRG1 is differentially expressed in cancerous tissue compared with normal testicular tissue and demonstrates a unique expression pattern in normal testes; therefore, this gene may be involved in the occurrence and development of testicular germ cell tumors (TGCT). In the present study, the expression level of TDRG1 was downregulated in human TGCT NTERA-2 cells by RNA interference (RNAi) in order to investigate the association between TDRG1 and TGCT. The TDRG1 mRNA and protein expression levels in NTERA-2 cells were significantly inhibited following transfection with specific RNAi plasmids. The ability to proliferate (inhibited by 15.4% at day 3 and 26.1% at day 5; P<0.001) and invade (reduced by 49.1%; P=0.01) in vitro was suppressed in cells in which the expression level of TDRG1 was reduced, and a corresponding increase in the apoptotic potential was observed (the early apoptotic potential and total apoptotic potential were increased by 75%; P=0.019 and 54.8%; P=0.009, respectively). The results of the present study indicated that the biological behavior of NTERA-2 cells is associated with TDRG1 expression levels, and that this gene may be a novel target candidate in the treatment of TGCT.

Knockdown of HMGN5 suppresses the viability and invasion of human urothelial bladder cancer 5637 cells in vitro and in vivo.

The high-mobility group nucleosome-binding domain 5 (HMGN5) is a new and typical member of HMGN protein family. Numerous studies confirmed that HMGN5 was highly expressed in several kinds of malignant tumors, but its role in cancer progression of urothelial bladder cancer (UBC) has not been fully clarified. This study aimed to further investigate the oncogenic role of HMGN5 in UBC 5637 cells employing in vitro and in vivo models and to explore the mechanism [corrected].RNA interference was used to down-regulate HMGN5 expression in 5637 cells by a shRNA expression lentiviral vector. Then cell viability, apoptosis and cell cycle distribution, invasion were detected by MTT assay, flow cytometry and transwell assay, respectively. Tumor growth was also evaluated in nude mice. As a result, successful transfection was confirmed using fluorescence microscopy and HMGN5 was efficiently inhibited. HMGN5 knockdown suppressed invasion, and induced G1/S cell cycle arrestbut not apoptosis and thus contributed to decreased cell viability in UBC 5637 cells [corrected]. Consistent with the cell cycle arrest, the protein expression levels of cyclin D1 were decreased. In vivo study further showed that HMGN5 knockdown affected the tumorigenesis of 5637 cells in nude mice. Western blot also demonstrated that the expression of E-cadherin was enhanced, while the expression of VEGF-C was decreased in 5637 cells depleted of HMGN5 [corrected].In conclusion, we provide both in vivo and in vitro evidence that HMGN5 contribute to the growth and invasion of UBC 5637 cell line and HMGN5 could be exploited as a target for therapy in UBC.