The complex and heterogeneous nature of hepatocellular carcinoma (HCC) hampers the identification of effective therapeutic strategies. Cancer stem cells (CSCs) represent a fraction of cells within tumors with the ability to self-renew and differentiate, and thus significantly contribute to the formation and maintenance of heterogeneous tumor mass. Increasing evidence indicates high plasticity in tumor cells, suggesting that non-CSCs could acquire stem cell properties through de-differentiation or reprogramming processes. In this paper, we reveal KLF4 as a transcription factor that can induce a CSC-like phenotype in non-CSCs through upregulating the EpCAM and E-CAD expression. Our studies indicated that KLF4 could directly bind to the promoter of EpCAM and increase the number of EpCAM⁺/CD133⁺ liver cancer stem cells (LCSCs) in the HuH7 HCC cell line. When KLF4 was overexpressed in EpCAM⁻/CD133⁻ non-stem cells, the expressions of hepatic stem/progenitor cell genes such as CK19, EpCAM and LGR5 were significantly increased. KLF4 overexpressing non-stem cells exhibited greater cell viability upon sorafenib treatment, while the cell migration and invasion capabilities of these cells were suppressed. Importantly, we detected an increased membranous expression and colocalization of β-CAT, E-CAD and EpCAM in the KLF4-overexpressing EpCAM⁻/CD133⁻ non-stem cells, suggesting that this complex might be required for the cancer stem cell phenotype. Moreover, our in vivo xenograft studies demonstrated that with a KLF4 overexpression, EpCAM⁻/CD133⁻ non-stem cells attained an in vivo tumor forming ability comparable to EpCAM⁺/CD133⁺ LCSCs, and the tumor specimens from KLF4-overexpressing xenografts had increased levels of both the KLF4 and EpCAM proteins. Additionally, we identified a correlation between the KLF4 and EpCAM protein expressions in human HCC tissues independent of the tumor stage and differentiation status. Collectively, our data suggest a novel function for KLF4 in modulating the de-differentiation of tumor cells and the induction of EpCAM⁺/CD133⁺ LCSCs in HuH7 HCC cells.

Testicular torsion is twisting of the spermatic cord around its axis, which impairs blood flow and causes ischemia and formation of free radicals. Ferulic acid is a phenolic acid of the hydroxycinnamic family that is found in the seeds and leaves of plants; it is present in substantial amounts in fruits and vegetables. We investigated the protective effect of ferulic acid on experimental testicular torsion in rats. Animals were divided randomly into five groups: control, ethyl alcohol, torsion, torsion-detorsion, and torsion-detorsion + ferulic acid. Histopathology was assessed using hematoxylin and eosin, and periodic acid-Schiff staining. Tissues were assessed using TUNEL, active caspase-3, myeloperoxidase and inducible nitric oxide synthase immunostaining. Biochemical changes were assessed using assays for superoxide dismutase, malondialdehyde, glutathione peroxidase and glutathione. Ferulic acid reduced the levels of free radicals and increased the levels of antioxidants. Ferulic acid also reduced histopathological changes and germ cell differentiation in the testis following torsion-detorsion. Ferulic acid should be investigated further as a potential treatment for sequelae of torsion-detorsion injury.

Background and objectives: It has been shown that electromagnetic fields (EMFs) have negative effects on the reproductive system. The biological effects of EMF on the male reproductive system are controversial and vary depending on the frequency and exposure time. Although a limited number of studies have focused on the structural and functional effects of EMF, the effects of prenatal and postnatal EMF exposure on testes are not clear. We aimed to investigate the effects of 50-Hz, 3-mT EMF exposure (5 days/wk, 4 h/day) during pre- and postnatal periods on testis development. Materials and Methods: Pups from three groups of Sprague-Dawley pregnant rats were used: Sham, EMF-28 (EMF-exposure applied during pregnancy and until postnatal day 28), EMF-42 (EMF-exposure applied during pregnancy and until postnatal day 42). The testis tissues and blood samples of male offspring were collected on the postnatal day 42. Results: Morphometric analyses showed a decrease in seminiferous tubule diameter as a result of testicular degeneration in the EMF-42 group. Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were decreased in the EMF-42 group. Lipid peroxidation levels were increased in both EMF groups, while antioxidant levels were decreased only in the EMF-28 group. We found decreased levels of vascular endothelial growth factor (VEGF) and insulin-like growth factor-1 (IGF1) in the EMF-42 group, and decreased levels of the SRC homology 3 (SH3) and multiple ankyrin repeat domain (SHANK3) in the EMF-28 group in the testis tissue. Conclusions: EMF exposure during pre- and postnatal periods may cause deterioration in the structure and function of testis and decrease in growing factors that would affect testicular functions in male rat pups. In addition to the oxidative stress observed in testis, decreased SHANK3, VEGF, and IGF1 protein levels suggests that these proteins may be mediators in testis affected by EMF exposure. This study shows that EMF exposure during embryonic development and adolescence can cause apoptosis and structural changes in the testis.

Purpose: Ischemic priapism (IP) is characterised by prolonged, painful erections that require urgent intervention to prevent corporal fibrosis and erectile dysfunction. Hydrogen sulfide (H₂S), a gaseous mediator, has been shown to exert protective effects against ischemia-reperfusion (IR) injury by reducing oxidative stress, enhancing mitochondrial function, and inhibiting apoptosis. This study aimed to investigate the role of H₂S during IP and reperfusion and its potential protective effects against penile tissue injury. Methods: Twenty-eight male Wistar rats were divided into four groups: control, IP, IP-R, and IP-[NaHS]-R. The control group underwent penectomy only. In the IP group, penectomy was performed after 4 h of priapism, whereas in the IP-R group, it was performed after 4 h of priapism followed by 1 h of reperfusion. In the IP-[NaHS]-R group, rats received NaHS (75 µmol/kg, intraperitoneally) 10 min before reperfusion. Endogenous H₂S levels were determined by the methylene blue assay, and hypoxia-inducible factor-1α (HIF-1α) expression was evaluated by immunohistochemistry. Results: Histopathological analyses were performed to assess tissue alterations. The IP and IP-R groups showed edema, inflammation, desquamation, vasocongestion, and increased collagen deposition, and exhibited reduced H₂S levels ( p < 0.05) and elevated HIF-1α ( p < 0.0001) compared with control group. NaHS administration increased H₂S levels, reduced HIF-1α expression, and alleviated histopathologic changes. Conclusion: Decreased H₂S levels during ischemia and reperfusion were associated with penile tissue injury in IP. NaHS mitigated morphological damage during the initial phase of reperfusion, indicating the potential involvement of H₂S signaling in reperfusion-related corporal tissue injury. However, endpoints related to the resolution of priapism, such as detumescence time and intracavernosal pressure (ICP), have not been evaluated. These assessments are necessary to determine the translational safety and therapeutic applicability of these treatments in cases of IP.