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.

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.

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.

[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.

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.

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.

[Comparison the effects of prone and modified recumbent positions on minimal invasive percutaneous nephrolithotomy].

OBJECTIVE: To compare the safety and efficacy of prone and modified recumbent positions on minimal invasive percutaneous nephrolithotomy. METHODS: A total of 62 patients with upper urinary calculi were grouped into two groups, one of which consisted of 27 patients who underwent the minimal invasive percutaneous nephrolithotomy with modified recumbent position, and the other 35 patients with prone position. There was no significant statistical difference in the age, gender and complications between the two groups before surgery (P>0.05). Duration of and blood loss during surgery, complications in the perioperative period, and the length of postoperative hospital stay were all recorded. The data were analyzed by SPSS 13.0. RESULTS: Surgery was successful in all cases. There was no failure to puncture nor need to resort to open surgery. Average operation duration for the modified recumbent position group was (85.1± 25.3) min vs (97.2±30.6) min for the prone position group. Mean blood loss during the operation was (117.5± 49.7) mL vs (149.3±53.1) mL. There were no severe complications during and after surgery in the modified recumbent position group. In the prone position group, s one patient suffered pneumothorax during the operation and two suffered selective renal artery embolization because of massive hemorrhaging following the operation. There were significant differences in blood loss during surgery, in complications during the perioperative period, and in length of postoperative stay in hospital (P<0.05) between the two groups. CONCLUSION: The patients are safer and more easily tolerate the minimal invasive percutaneous nephrolithotomy in the modified recumbent position than in the prone position, though the treatment efficacy of these two kinds of operation is similar. It is recommended that the modified recumbent position should be used generally in the percutaneous nephrolithotomy.

[Construction of TDRG1 shRNA expression vector and interfering effect of TDRG1 shRNA expression vector on NTERA-2 cells].

OBJECTIVE: To construct short hairpin RNA interfering expression vector of TDRG1,and detect the specific interfering effect of TDRG1-shRNA expression vector on NTERA-2 cells. METHODS: Oligos for short hairpin RNA targefing for TDRG1 were designed and connected to the expression vector pGPU6/GFP/Neo to construct the TDRG1 shRNA expression vector. The recombinant plasmid TDRG1-shRNA486, TDRG1-shRNA738, TDRG1-shRNA921 and lipofectamine ™2000 were used to generate and transfect shRNA into NTERA-2 cells. Expression of TDRG1 mRNA was assayed by RT-PCR. RESULTS: TDRG1-shRNA expression vector was successfully constructed. TDRG1-shRNA486 was more effective in the suppression of TDRG1 with significant reduction of TDRG1 mRNA. CONCLUSION: TDRG1-shRNA can interfere the expression of TDRG1 in NTERA-2 cells.

[The applied research of a neotype medical drainage bag in clinical care].

OBJECTIVE: To compare and evaluate the effectiveness of two kinds of medical drainage bag. METHODS: 206 patients were randomly divided into two groups each of which consisted of 103 patients. All the data including four indices, such as the time required to replace the drainage bags, the incidence of the bags detached, draining fluid splashing rates during the replacement of the bags, patient and medical staff satisfaction, were collected and analyzed statistically. RESULTS: The time required to replace the drainage bags, the incidence of the bags detached and draining fluid splashing rates during the replacement of the bags of the experimental group were significantly lower than those of the control group (P < 0.05), while the patient and medical staff satisfaction were significantly higher than those of the control group (P < 0.05). CONCLUSION: It is convenient, quick and time and effort saving to use the neotype medical drainage bags. Hence, the use of neotype medical drainage bags could help to improve the work efficiency, effectively prevent occupational injuries and protect health care workers.

Characterization of a novel human testis-specific gene: testis developmental related gene 1 (TDRG1).

Spermatogenesis is a highly coordinated physiological process that requires the correct expression and functions of thousands of developmentally regulated genes. The regulation of spermatogenesis is not well defined, since majority of the related genes have neither been identified nor fully characterized. Hence, it is meaningful to identify and characterize these genes to reveal the mechanism underlying spermatogenesis. In this study, using digital differential display, we identified a novel human testis-specific gene, testis developmental related gene 1 (TDRG1, GenBank DQ168992), via electronic subtraction of human testis UniGene databases from those of non-reproductive tissues. The transcript of the TDRG1 gene has an open-reading frame that encodes 100 amino acids. We next prepared the anti-TDRG1 monoclonal antibody 10B6 and confirmed that it specifically recognizes an 11-kDa protein in the tissue extracts from an adult human testicular sample (age 31 years) by Western blot analysis. RT-PCR coupled with immunohistochemistry of human tissues demonstrated that TDRG1 is exclusively expressed in the testis but not in any other non-reproductive tissues. TDRG1 is mainly located in spermatogenic cells in seminiferous tubules of adult testis. Furthermore, TDRG1 shows the highest expression level in human post-puberty testis, with the expression levels decreasing afterwards with aging. Importantly, TDRG1 mRNA is undetectable in the fetal testis, as judged by RT-PCR. In conclusion, TDRG1 is a developmentally regulated testicular-specific gene. We suggest that TDRG1, a newly identified testis-specific gene, may play important roles in human spermatogenesis.

[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.

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.

[Preparation and identification of monoclonal antibody against human testis development related gene 1].

OBJECTIVE: To construct a prokaryotic plasmid to express the testis development related gene 1 (TDRG1) recombinant protein and obtain anti-TDRG1 mAb by immunizing mice, and to identify the biological properties of the mAb. METHODS: The coding sequence of TDRG1 was amplified by RT-PCR from normal human testis tissue and cloned into the vector pET21, and then was expressed in the E. coli BL 21(DE3) to get TDRG1 recombinant protein. The purified TDRG1 recombinant protein was injected to immunize the BALB/C mice to develop anti-TDRG1 mAb. Splenocytes of the immunized mice were collected and fused with the mouse myeloma cell line Sp2/0 cells. The hybridoma cells that secreted anti-TDRG1 mAb were subcloned with limited dilution. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate titers and subtypes of mAb. Western blot and immunohistochemistry were used to detect specificity of mAb. RESULTS: The prokaryotic plasmid expressing the recombinant protein was constructed, and the TDRG1 recombinant protein was expressed and purified. Two hybridoma cell lines that secreted anti-TDRG1 mAb were obtained. The titer of the mAb in ascites was 1 : 1.6 x 10(6), and the subtype of the mAb was IgG(1). Westem blot and immunohistochemistry analysis indicated the mAb showed specific combination with TDRG1 protein in human testes. CONCLUSION: The TDRG1 recombinant protein is highly purified and has strong antigenicity. The anti-TDRG1 mAb is produced successfully.

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.

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.

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.