Yunpeng Zhu

Background: Macrophages are highly plastic cells, and macrophage-derived exosomes (M-Exos) have been implicated in inflammation-related pathophysiologies, such as tissue injury and fibrosis repair. This study aimed to investigate the possible effects of M-Exos on the initiation and development of urethral fibrosis and stricture after injury, and to elucidate the underlying mechanisms. Methods: In this study, we used time-tracking immunofluorescence staining for M1 and M2 macrophage markers to characterize sequential properties in the site of injured urethra. Next, we harvested these exosomes from different macrophages to co-culture with fibroblasts to further confirm the role of exosome-mediated M2 macrophage-fibroblast communication. Then, high-throughput micro-RNA (miRNA) sequencing was performed to detect the candidate exosomal miRNA and its target gene. Furthermore, fibroblasts were transfected with mRFP-GFP-LC3 plasmid to detect the autophagy role of SIRT1 in the urethral fibroblasts fibrogenesis. Results: Here we found that M2-polarized macrophages triggered and dominated the fibrotic scene, purified exosomes from M2 macrophages exacerbated urethral fibroblast fibrogenesis, and the inhibition of exosome secretion abolished fibroblast fibrogenesis. Moreover, miR-34a-5p, which is highly enriched and packaged within M2-Exos, can be transferred from M2 macrophages into urethral fibroblasts, resulting in deterioration of proliferation and fibrogenesis. Mechanistically, M2-Exos miR-34a-5p could directly interact with the 3ʹ-UTR of SIRT1, thereby suppressing SIRT1 expression in fibroblasts, leading to the blockage of autophagosome-lysosome fusion to impair urethral fibroblast autophagy flux and further exacerbate fibrogenesis. More importantly, repression of miR-34a-5p in M2-Exos mitigated-urethral strictures in rats with damaged urethra. Conclusion: M2 macrophage-derived exosomes miR-34a-5p could aggravate the development of urethral fibrosis and stricture by blocking autophagosome-lysosome fusion in urethral fibroblasts and further accelerating fibrogenesis by directly targeting SIRT1, suggesting that M2-Exo miR-34a-5p and SIRT1 could serve as promising therapeutic targets for urethral stricture.

The perplexing issues related to positive surgical margins and the considerable negative consequences associated with systemic chemotherapy have posed ongoing challenges for clinicians, especially when it comes to addressing bladder cancer treatment. The current investigation describes the production of nanocomposites loaded with gemcitabine (GEM) and cisplatin (CDDP) through the utilization of electrospinning technology. In vitro and in vivo studies have provided evidence of the strong effectiveness in suppressing tumor advancement while simultaneously reducing the accumulation of chemotherapy drugs within liver and kidney tissues. Mechanically, the GEM and CDDP-loaded electrospun nanocomposites could effectively eliminate myeloid-derived suppressor cells (MDSCs) in tumor tissues, and recruit CD8⁺ T cells and NKp46⁺ NK cells to kill tumor cells, which can also effectively inhibit tumor microvascular formation. Our investigation into the impact of localized administration of chemotherapy through GEM and CDDP-loaded electrospun nanocomposites on the tumor microenvironment will offer novel insights for tackling tumors. Graphical

Objective. To explore whether Stromal Interaction Molecule‐1 (STIM1) participates in the phenotypic transformation of NRK‐52E cells under high‐calcium microenvironment. Materials and Methods. NRK‐52E cells were treated with high concentration of calcium. The viability and apoptosis of cells were detected by CCK‐8 (cell counting kit‐8) and flow cytometry, respectively. The expression changes of phenotypic marker proteins (E‐cadherin and OPN) and calcium channel proteins (STIMl and Orai1) in high‐calcium environment were detected by western blotting and real‐time quantitative polymerase chain reaction. The expression of STIMl protein in NRK‐52E cells was upregulated and downregulated by plasmid‐STIM1 and plasmid‐shRNA‐STIMl, respectively. The expressions of phenotypic marker proteins after upregulation or downregulation of STIMl were detected again. Besides, the intracellular calcium concentrations of NRK‐52E cells in different treatments were detected by flow cytometry. Results. High‐calcium microenvironment can promote the phenotypic transformation and the adhesion of calcium salts in NRK‐52E cells and simultaneously suppress the expression of STIMl protein in NRK‐52E cells. Downregulation of STIMl protein could also promote the phenotype transformation, while both the gene silence of matrix gla protein (MGP) and overexpression of STIMl showed reverse results. Conclusion. STIMl protein plays an important role in promoting phenotypic transformation of NRK‐52E cells in high‐calcium microenvironment.

The positive surgical margins in solid tumors has been a disturbing issue for clinicians. Chemotherapy is an important method to deal with the positive margin. However, systemic chemotherapy is required for long-term administration and has great side effects on health, which cause great pain to the patients. Local administration of slow-release materials provides an opportunity to improve the situation. In this study, we utilized electrospinning technology to create the drug sustained-release materials with nanofibrous structure, which were made from polylactic acid and a certain proportion of chemotherapy drugs (gemcitabine and cisplatin). In vitro release behavior of the drug sustained-release materials were explored by the high-performance liquid chromatography. The antitumor efficacy of the drug sustained-release materials was preliminarily verified in prostate cancer and breast cancer in vitro . Through animal models of breast cancer, the drug sustained-release materials in the treatment of the positive margin has been well documented in vivo , and we also found that the drug sustained-release materials could definitely reduce the liver damage and myelosuppression compared with systemic chemotherapy. In summary, the experimental results showed that the local administration of the drug sustained-release materials could effectively inhibit the growth of the positive incision margins and definitely reduce the partial side effects associated with systemic chemotherapy.

Background: Second primary malignancy (SPM) challenges survival and surveillance protocols among renal cell carcinoma (RCC) survivors. The incidence, temporal patterns, survival outcomes, and risk factors of SPM after T1-4N0-1M0 RCC diagnosis need to be investigated. Method: A nested case-control study that was designed using the Surveillance, Epidemiology, and End Results database from 2004-15; A cohort of 6204 SPM were matched with a control group of 37224 non-SPM. Results: SPM shortens the overall survival (hazard ratio [HR], 1.34; 95% confidence interval [CI]: 1.28-1.42, P< 0.001). The median time interval to SPM was 54.5 months. The adjusted standardized incidence ratio (SIR) of SPM increases by survival time (SIR_12~35-month: 12.04; SIR_36~59-month: 12.67; SIR_60~19-month: 16.08; SIR_120+-years: 25.01, all P< 0.001), and decreased with age (SIR_18~44-years: 86.68; SIR_45~59-years: 26.95; SIR_60~74-years: 12.43; SIR_75+-years: 10.66, all P< 0.001). The second primary RCC onset, especially contralateral kidney, has the highest SIR (SIR: 54.6; 95%CI: 51.0~58.4) among all sites of SPM. Prostate cancer (29.8%) in male and breast cancer (23.5%) in female were the most common SPM site. Older age, black race, male gender, higher family income statues, papillary RCC, and lower TNM stage significantly increases the risk of SPMs diagnosis. A longer time to SPM interval positively associated with a higher tumor stage of a SPM onset (P _trend Conclusion: Collectively, our study described the epidemiological characteristics of SPM among RCC survivors and identified the independent predictors of the SPM onset and its survival outcomes, which provides the clinicians for patients consulting and long-term individual-, tailored site-, and time-specific surveillance to improve survival outcomes.

Calcium oxalate (CaOx) is the most common type of urinary stone. Increase of ROS and NADPH oxidase gives rise to inflammation and injury of renal tubular cells, which promotes CaOx stone formation. Recent studies have revealed that the renin‐angiotensin system might play a role in kidney crystallization and ROS production. Here, we investigated the involvement of Ang II/AT1R and losartan in CaOx stone formation. NRK‐52E cells were incubated with CaOx crystals, and glyoxylic acid‐induced hyperoxaluric rats were treated with losartan. Oxidative stress statuses were evaluated by detection of ROS, oxidative products (8‐OHdG and MDA), and antioxidant enzymes (SOD and CAT). Expression of NADPH oxidase subunits (Nox2 and Nox4), NF‐κB pathway subunits (p50 and p65), and stone‐related proteins such as OPN, CD44, and MCP‐1 was determined by Western blotting. The results revealed upregulation of Ang II/AT1R by CaOx treatment. CaOx‐induced ROS and stone‐related protein upregulation were mediated by the Ang II/AT1R signaling pathway. Losartan ameliorated renal tubular cell expression of stone‐related proteins and renal crystallization by inhibiting NADPH oxidase and oxidative stress. We conclude that losartan might be a promising preventive and therapeutic candidate for hyperoxaluria nephrolithiasis.

Many studies compared the serum/plasma 1,25 dihydroxyvitamin D₃ (1,25(OH)₂D) and 25 hydroxyvitamin D₃ (25(OH)D) between people with and without nephrolithiasis, and their results were conflicting. After systematically searching PubMed, Web of Science, The Cochrane Library, CNKI, and the Wanfang Database, we conducted a meta-analysis. Thirty-two observational studies involving 23,228 participants were included. Meta-analysis of these studies showed that of stone formers (SFs), calcium SFs had significantly higher concentrations of 1,25(OH)₂D (weighted mean difference (WMD), 10.19 pg/mL; 95% confidence interval (CI), 4.31–16.07; p = 0.0007 and WMD, 11.28 pg/mL; 95% CI, 4.07–18.50; p = 0.002, respectively) than non-stone formers, while the levels of 25(OH)D (WMD, 0.88 ng/mL; 95% CI, −1.04–2.80; p = 0.37 and WMD, −0.63 ng/mL; 95% CI, −2.72–1.47; p = 0.56, respectively) are similar. Compared with controls and normocalciuria SFs, hypercalciuria SFs had increased circulating 1,25(OH)₂D (WMD, 9.41 pg/mL; 95% CI, 0.15–18.67; p = 0.05 and WMD, 2.75 pg/mL; 95% CI, −0.20–5.69; p = 0.07, respectively) and markedly higher 25(OH)D (WMD, 5.02 ng/mL; 95% CI, 0.99–9.06; p = 0.01 and WMD, 5.02 ng/mL; 95% CI, 2.14–7.90; p = 0.0006, respectively). Normocalciuria SFs had elevated 1,25(OH)₂D level (WMD, 6.85 pg/mL; 95% CI, −5.00–18.71; p = 0.26) and comparable 25(OH)D (WMD, 0.94 ng/mL; 95% CI, −3.55–5.43; p = 0.68). Sensitivity analysis generated similar results. Current evidence suggests that increased circulating 1,25(OH)₂D is associated with urinary stones and a higher level of circulating 25(OH)D is significantly associated with hypercalciuria urolithiasis. Further studies are still needed to reconfirm and clarify the role of vitamin D in the pathogenesis of stones.