Regulation of Spontaneous Contractions in Intact Rat Bladder Strips and the Effects of Hydrogen Peroxide

Enhanced spontaneous contractions are associated with overactive bladder. Elevated levels of reactive oxygen species might contribute to enhanced spontaneous contractions. We investigated the regulation of spontaneous contractions and the effects of hydrogen peroxide (H2O2) in intact rat bladder strips. The spontaneous contractions were measured using a tissue bath system. The vehicle or the specific activators/blockers were applied and followed by the application of 0.003 g% H2O2. The basal tension, amplitude, and frequency of spontaneous contractions were quantified. Nisoldipine and bisindolylmaleimide 1 had no effects on spontaneous contractions. SKF96365 and Y27632 decreased basal tension and amplitude. Ryanodine slightly increased frequency. Both iberiotoxin and NS-1619 increased amplitude. Apamin reduced frequency but increased amplitude. NS-309 inhibited both the amplitude and frequency. The basal tension and amplitude increased when H2O2 was applied. Pretreatment with NS-309 inhibited H2O2-elicited augmented amplitude and frequency, while pretreatment with Y-27632 inhibited the augmented basal tension. The combined application of NS-309 and Y27632 almost eliminated spontaneous contractions and its augmentation induced by H2O2. In conclusion, Ca2+ influx, Rho kinase activation, and SK channel inactivation play important roles in spontaneous contractions in intact bladder strips, whereas only latter two mechanisms may be involved in H2O2-elicited increased spontaneous contractions

MicroRNA-17, 20a Regulates the Proangiogenic Function of Tumor-Associated Macrophages via Targeting Hypoxia-Inducible Factor 2α

<div><p>Tumor-associated macrophages (TAMs) constitute a major component of the leukocyte infiltrate of most solid tumors, and they usually exhibit a proangiogenic phenotype which facilitates tumor growth in most circumstances. However, the precise mechanisms regulating the proangiogenic properties of TAMs remain largely unclear. In the present study, we found that the expression of hypoxia-inducible factor 2α (HIF-2α) was significantly up-regulated in macrophages from tumor tissues of several solid tumors. Macrophages exposed to tumor cell line derived-culture supernatants (TSN) also expressed high levels of HIF-2α in vitro, without a requirement for hypoxia. We identified miR-17 and miR-20a as the key regulators of HIF-2α expression in TAMs, and autocrine IL-6 played an important role in mediating the expression of miR-17, miR-20a, and thereafter HIF-2α in TAMs. Furthermore, the elevated HIF-2α in TAMs stimulated transcription of a set of proangiogenic genes such as VEGFA and PDGFB, which might in turn contribute to the angiogenic process within tumors. Our data provide evidence in support of the critical role of HIF-2α in the proangiogenic activity of TAMs and also reveal a novel mechanism by which miRNAs regulate TAM functions through modulation of HIF-2α expression under non-hypoxic conditions.</p></div

Blood-based liquid biopsy: insights into early detection, prediction, and treatment monitoring of bladder cancer

Abstract Bladder cancer (BC) is a clinical challenge worldwide with late clinical presentation, poor prognosis, and low survival rates. Traditional cystoscopy and tissue biopsy are routine methods for the diagnosis, prognosis, and monitoring of BC. However, due to the heterogeneity and limitations of tumors, such as aggressiveness, high cost, and limited applicability of longitudinal surveillance, the identification of tumor markers has attracted significant attention in BC. Over the past decade, liquid biopsies (e.g., blood) have proven to be highly efficient methods for the discovery of BC biomarkers. This noninvasive sampling method is used to analyze unique tumor components released into the peripheral circulation and allows serial sampling and longitudinal monitoring of tumor progression. Several liquid biopsy biomarkers are being extensively studied and have shown promising results in clinical applications of BC, including early detection, detection of microscopic residual disease, prediction of recurrence, and response to therapy. Therefore, in this review, we aim to provide an update on various novel blood-based liquid biopsy markers and review the advantages and current limitations of liquid biopsy in BC therapy. The role of blood-based circulating tumor cells, circulating tumor DNA, cell-free RNA, exosomes, metabolomics, and proteomics in diagnosis, prognosis, and treatment monitoring, and their applicability to the personalized management of BC, are highlighted

Study on Heat Storage Performance of Phase Change Reservoir in Underground Protection Engineering

In view of the main problems of the condensing heat discharge modes of the existing underground air-conditioning system, the technical scheme of using phase change heat storage modules to improve the heat storage capacity of the reservoir is proposed. By establishing a 3D flow and transient heat transfer model of the phase change reservoir, the effects of thermal property parameters, package size and arrangement of the phase change heat storage modules on the heat storage performance of the phase change reservoir were quantitatively analyzed based on three indexes: heat storage capacity per volume &Delta;q, guaranteed efficiency coefficient &eta; and slope of temperature rise per unit load &epsilon;. The results show that when the phase change temperature is 29 &deg;C (23 &deg;C increased to 33 &deg;C) and the latent heat value is 250 kJ/kg (100 kJ/kg increased to 250 kJ/kg), &Delta;q (110.92 MJ/m3, 112.83 MJ/m3) and &eta; (1.22, 1.24) under both conditions are at their most, respectively, indicating that the phase change temperature should be less than 4 &deg;C at the outlet temperature of the reservoir, and phase change materials with a high latent heat should be selected in engineering design whenever possible. When the size of the phase change module is 150 mm &times; 20 mm and the phase change reservoir adopts four intakes, &epsilon; (0.259, 0.244) under both conditions is the smallest, indicating that increasing the area of the phase change heat storage module and the fluid and increasing the inlet disturbance of the reservoir can enhance its heat storage capacity

Role of IL-6 in downregulation of miR-17 and miR-20a and upregulation of HIF-2α expression in TAMs.

<p>(A) Level of IL-6 mRNA in CD14⁺ cells isolated from non-tumoral (Non-) or intratumoral (Intra-) tissue of seven HCC patients was analyzed by qPCR. Results are presented as fold-changes. (B) Healthy PBMC-derived CD14⁺ cells were treated with medium alone or HepG2 TSN for 24h. IL-6 concentrations in the culture supernatants were determined by ELISA. (C, D, E) Healthy PBMC-derived CD14⁺ cells were treated HepG2 TSN and transfected with NC or si-IL-6R (C). Healthy PBMC-derived CD14⁺ cells were treated with medium, TCM, TCM with anti-IL-6, or TCM with IgG control for 7 days (D). Healthy PBMC-derived CD14⁺ cells were treated with medium alone or recombinant IL-6 for 7 days (E). MiR-17 and miR-20a in these cells were analyzed by qPCR. Expression of HIF-2α was determined by Western blotting. qPCR data were presented as fold-changes and normalized to ACTB for mRNA expression or RNU6B for miRNAs expression. Values represent the mean ± SEM for A and mean ± SD for the others. All data shown are representative of at least four separate experiments. *<i>P</i><0.05, **<i>P</i><0.01 compared with the indicated groups.</p

Upregulation of HIF-2α expression in TAMs.

<p>(A) Adjacent sections of paraffin-embedded HCC tissue (n = 9) were stained with an anti-HIF-2α or anti-CD68 antibody. (B) HIF-2α expression in macrophages from frozen sections of HCC tissue (n = 6) was analyzed by confocal microscopy. HIF-2α, red; CD68, green; DAPI, blue. (C) Level of HIF-2α protein in CD14⁺ cells isolated from non-tumoral (Non-) or intratumoral (Intra-) tissue of 4 HCC patients was analyzed by immunoblotting. (D) Healthy PBMC-derived monocytes were treated with medium alone (Med) for 7 days, or with HepG2 TSN under normoxic condition (HepG2 TSN) for 7 days, or with medium alone for 6 days and then exposed to 1% O₂ for another 24 h (hypoxia). Expression of HIF-2α and HIF-1α was determined by Western blotting. This data shown are representative of four separate experiments.</p

HIF-2α modulates proangiogenic gene expression in TAMs.

<p>(A) CD14⁺ cells were isolated from non-tumoral (Non-) or intratumoral (Intra-) tissue of seven HCC patients. (B) Healthy PBMC-derived CD14⁺ cells were treated with medium alone or HepG2 TSN for 7 days. (C, D) Healthy PBMC-derived CD14⁺ cells were treated with HepG2 TSN and transfected with NC, si-HIF-2α (C), or miR-17 and miR-20a mixture (D). Levels of VEGFA and PDGFB mRNA expression in these cells were analyzed by qPCR. Upper panel of C shows the knockdown efficiency of si-HIF-2α. All results are normalized to ACTB and presented as fold-changes. All data shown are representative of at least four separate experiments. *<i>P</i><0.05, **<i>P</i><0.01 compared with the indicated groups.</p

Novel agents and clinical trials in castration-resistant prostate cancer: latest updates from 2023 ASCO-GU Cancers Symposium

Abstract Numerous novel and effective therapeutic agents and clinical trials addressing castration-resistant prostate cancer (CRPC) were reported during the 2023 American Society of Clinical Oncology-Genitourinary (ASCO-GU) Cancers Symposium. Notably, radionuclide drug conjugates (RDC), specifically 177Lu/111In-J591 and 225Ac-J591, exhibited enhanced therapeutic efficacy in treating patients with CRPC. Furthermore, promising treatment approaches for CRPC included dual anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and anti-programmed death-1 (PD-1) blockade in rare tumors (DART)-Lorigerlimab, prostate stem cell antigen (PSCA)-directed chimeric antigen receptor (CAR)-T cell immunotherapy-BPX-601, and protein kinase inhibitor (AKTi)-CAPltello-280. We have summarized the latest CRPC treatment strategies presented at the 2023 ASCO-GU Cancers Symposium, along with recent advances in CRPC clinical trials

MiR-17 and miR-20a regulate HIF-2α expression at the post-transcriptional level in TAMs.

<p>(A, B) CD14⁺ cells were isolated from non-tumoral (Non-) or intratumoral (Intra-) tissues of seven HCC patients (A). Healthy PBMC-derived CD14⁺ cells were treated with medium alone or HepG2 TSN for 7 days (B). Levels of HIF-2α mRNA, miR-17 and miR-20a in these cells were analyzed by qPCR. Data were presented as relative level and normalized to ACTB for mRNA expression or RNU6B for miRNAs expression. (C) Pairing of miR-17 and miR-20a with three HIF-2α 3′UTR regions (nucleotides 170–176, 816–822 and 1500–1506 of human HIF-2α 3′UTR), and mutations introduced into the luciferase reporter construct are shown. (D, E) Luciferase activity of HepG2 cells transfected with the indicated reporters was analyzed. pRL-CMV expressing <i>Renilla</i> luciferase was cotransfected as an internal control and the luciferase activity of each sample was normalized to this. The luciferase activity of the NC or anti-miR-C transfectant was assigned an arbitrary value of 1. (F) Healthy PBMC-derived CD14⁺ cells were treated with medium alone and then transfected with NC or anti-miR-17 and anti-miR-20a mixture (anti-miRNAs), or with HepG2 TSN and then transfected with NC or miR-17 and miR-20a mixture (miRNAs). HIF-2α expression was analyzed by Western blotting. Values represent the mean ± SEM for A and mean ± SD for the others. All data shown are representative of at least four separate experiments. *<i>P</i><0.05, **<i>P</i><0.01 compared with the indicated groups.</p

Model of the novel mechanism regulating HIF-2α expression under non-hypoxic condition within tumor microenvironment.

<p>Soluble factor(s) derived from tumor cells induced the secretion of IL-6 by stimulating its transcription. Autocrine IL-6 triggered the down-regulation of miR-17 and miR-20a, which in turn released the suppression of HIF-2α and led to its accumulation under non-hypoxic condition. HIF-2α further promoted the transcription of proangiogenic genes such as VEGFA and PDGFB and contributed to tumor angiogenesis.</p