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Objective To explore the dermal histological changes of rat expanded skin under persistent tissue hypoxic expansion and intermittent tissue hypoxic expansion. Methods From November 2016 to June 2017, a rat model was used in the Plastic Surgery Department of Xijing Hospital of the Air Force Medical University. After intermittent and continuous hypoxic tissue expansion, the skin samples of rats were stained with HE and Masson staining to observe the changes in the structure and collagen content of the dermis; Proliferating cell nuclear antigen (PCNA) and Terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining were used to determine the proliferation and apoptosis of dermal fibroblasts; CD31 staining was used to analyze the changes of vascular morphology and capillary density in the dermis. Results The dermal thicknesses in the IH group, PH group, and the normal group were（500.20±22.57）μm. (464.30±27.23）μm, and（759.40±27.23）μm respectively. The dermal thickness of the PH group was significantly smaller than the IH group (P<0.05). The collagen content in the IH group and PH group was significantly higher than that in the normal skin group (P<0.05). The densities of PCNA-positive fibroblasts in the dermis of the normal skin group, IH group, and PH group were 6.10 per high power field, 19.38 per high power field, and 15.82 per high power field respectively. The number of positive cells in the IH group was higher than that in the PH group (P<0.05). The number of TUNEL-positive fibroblasts in the dermis of the normal skin group, IH group, and PH group was 5.10 per high-power field, 16.05 per high-power field, and 13.05 per high-power field. The density of TUNEL-positive fibroblasts in the IH group was higher than that in the PH group, but no statistically significant difference was found between the two groups (P>0.05). The average numbers of capillaries per high-power visual field in the dermis of the IH group and PH group were 22.87±1.13 and 13.57± 0.43, a statistically significant difference was found between the two groups (P<0.05). Conclusion Intermittent hypoxic tissue expansion can more effectively stimulate the proliferation of dermal fibroblasts, reduce cell apoptosis, promote collagen synthesis, and reconstruct the dermal vascular network, which is more conducive to the biological growth of the expanded skin. [ABSTRACT FROM AUTHOR]

Simple Summary: It has remained obscure whether microglia proliferate in the diseased or injured brain. This study analyzed the mitotic ability of microglia in vivo in a rat facial nerve axotomy model in which the blood cells do not enter the parenchyma. Microglia were found to proliferate in the axotomized facial nucleus at 5–7 days post-insult. They enhanced the levels of macrophage colony-stimulating factor (M-CSF) as a mitotic factor, its receptor cFms, and cell-cycle-related proteins, such as proliferating cell nuclear antigen (PCNA) and cyclins in vivo. In the M-CSF-dependent proliferation system in vitro, c-Jun N-terminal kinase (JNK) and p38 were shown to function in the induction of PCNA/cyclins and cFms in microglia, respectively. These analyses revealed that some specific signaling cascades are linked to the proliferative reaction of microglia. Microglia in the axotomized facial nucleus were considered to exhibit a mitotic ability to rescue injured motoneurons as neurotrophic cells. Although microglia exist as a minor glial cell type in the normal state of the brain, they increase in number in response to various disorders and insults. However, it remains unclear whether microglia proliferate in the affected area, and the mechanism of the proliferation has long attracted the attention of researchers. We analyzed microglial mitosis using a facial nerve transection model in which the blood–brain barrier is left unimpaired when the nerves are axotomized. Our results showed that the levels of macrophage colony-stimulating factor (M-CSF), cFms (the receptor for M-CSF), cyclin A/D, and proliferating cell nuclear antigen (PCNA) were increased in microglia in the axotomized facial nucleus (axotFN). In vitro experiments revealed that M-CSF induced cFms, cyclin A/D, and PCNA in microglia, suggesting that microglia proliferate in response to M-CSF in vivo. In addition, M-CSF caused the activation of c-Jun N-terminal kinase (JNK) and p38, and the specific inhibitors of JNK and p38 arrested the microglial mitosis. JNK and p38 were shown to play roles in the induction of cyclins/PCNA and cFms, respectively. cFms was suggested to be induced through a signaling cascade of p38-mitogen- and stress-activated kinase-1 (MSK1)-cAMP-responsive element binding protein (CREB) and/or p38-activating transcription factor 2 (ATF2). Microglia proliferating in the axotFN are anticipated to serve as neuroprotective cells by supplying neurotrophic factors and/or scavenging excite toxins and reactive oxygen radicals. [ABSTRACT FROM AUTHOR]

Neuropeptide Y (NPY) is a widespread hormone in the central and peripheral nervous systems that maintains body homeostasis. Central actions of hypothalamic NPY have been identified in bone metabolism. Osteocytes are the main source of NPY in bone tissue, indicating that osteocytic NPY could be a local alternative pathway for hypothalamic mediated regulation of bone and bone cells. Here, we show that osteocytic NPY induces cell viability and proliferation. Osteocyte-derived factors are also closely associated with changes in cellular NPY mRNA levels. Furthermore, osteoblast mineralization was significantly induced by conditioned medium collected from NPYoverexpressing osteocytes (P < 0.05). Importantly, the NPY-AHNAK interaction was identified for the first time by co-immunoprecipitation, and significant inactivation of p-Smad1/5/9 was found in osteocytes with NPY or AHNAK insufficiency (P < 0.05). The activation of p-Smad1/5/9 reversed NPY insufficiency-caused decreases in the expression of osteocytic proliferating cell nuclear antigen and osteoblast markers including osteocalcin and Runx2 (P < 0.05); these findings showed an additional molecular mechanism by which NPY acts on cells through AHNAK-mediated Smad1/5/9 signalling. Collectively, our findings provide novel insights into the function of NPY in regulating osteocyte phenotype and function and provide new insights for further investigation into osteocytic NPY-mediated therapy. [ABSTRACT FROM AUTHOR]

To study the impact of aneuploid granulosa and stromal cells on folliculogenesis of small ovarian follicles from patients with mosaic Turner syndrome (TS) using a murine xenograft model. Laboratory study. University hospital. Ovarian cortical tissue was obtained by laparoscopic surgery from 18 patients with mosaic TS (aged 5–19 years) and 13 controls (aged 5–18 years). Part of each tissue fragment was used to karyotype ovarian cells in nongrafted tissue by fluorescence in situ hybridization. The remaining tissue was xenografted to severe combined immunodeficient mice for 5 months. Grafted tissue was analyzed for aneuploidy, and follicle density and morphology were determined. Expressions of proliferating cell nuclear antigen and anti-Müllerian hormone were investigated by immunohistochemistry. The impact of aneuploid granulosa and stromal cells on folliculogenesis. Fluorescence in situ hybridization of ovarian tissue before grafting was performed to determine the level of aneuploidy in stromal cells and oocytes and granulosa of small follicles. After xenografting, the level of aneuploidy of the newly formed layers of granulosa cells was again determined in secondary and antral follicles. Follicle density in ovarian tissue from patients with TS was significantly lower than in controls before grafting. Fluorescence in situ hybridization analysis confirmed that 101 of 104 oocytes from nongrafted tissue of patients with TS showed normal X chromosome content, whereas granulosa and stromal cells were mainly 45,X. Fragments from 12 patients with TS contained follicles at all stages after xenografting, including secondary and antral follicles. Follicle density in patients with TS and controls decreased significantly after grafting. Moreover, a shift from high to low proportions of 45,X granulosa cells was observed during folliculogenesis. Expression of proliferating cell nuclear antigen in follicles from patients with TS increased significantly during grafting. Secretion of anti-Müllerian hormone was impaired before grafting in peripubertal/postpubertal girls with TS, but recovered after grafting. Our study showed that small follicles from patients with mosaic TS undergoes folliculogenesis, despite the presence of aneuploid granulosa and stromal cells. Ovarian tissue cryopreservation could therefore be a valid option to preserve fertility in young patients with mosaic TS if sufficient numbers of follicles are present, thus preferably before the age of 12. [ABSTRACT FROM AUTHOR]

Recently, several studies have revealed that circular RNAs (circRNAs) play significant roles in various tumors, including lung adenocarcinoma (LUAD). Furthermore, it has been reported that circ_0005962 was upregulated in LUAD cells. Accordingly, this research is designed to investigate the mechanism of circ_0005962 on LUAD development. Circ_0005962, microRNA-3611 (miR-3611), and Cytochrome P450 family 24 subfamily A member 1 (CYP24A1) level were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation ability, cell cycle progression, and sphere formation ability were detected using Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), Colony formation, flow cytometry, and sphere formation assay. Protein levels of proliferating cell nuclear antigen (PCNA), Ki67, NANOG, CD133, OCT4, and CYP24A1 were determined using Western blot assay. Using bioinformatics software (Starbase3.0 and TargetScan), the binding between miR-3611 and circ_0005962 or CYP24A1 was predicted and proved using RNA Immunoprecipitation (RIP) and RNA pull-down assays. A xenograft tumor model in vivo was used to analyze the biological role of circ_0005962 on LUAD cell growth. Increased circ_0005962 and CYP24A1, and reduced miR-3611 were observed in LUAD tissues and cell lines. Functional assays testified that circ_0005962 depletion might hinder LUAD cell proliferation and sphere formation capability, but induced cell apoptosis in vitro. Molecular mechanism experiments exhibited that circ_0005962 served as a miR-3611 sponge and mediated CYP24A1 content by absorbing miR-3611. Additionally, silencing of circ_0005962 inhibited tumor growth in xenograft modes. Together, circ_0005962 was overexpressed in LUAD, and its deficiency repressed LUAD progression via targeting the miR-3611/ CYP24A1 axis, providing a novel mechanism for understanding the development of LUAD. [ABSTRACT FROM AUTHOR]

Ever‐increasing occurrence of plastic‐manufacturing industries leads to environmental pollution that has been associated with declined human health and increased incidence of compromised reproductive health. Female subfertility/infertility is a complex phenomenon and environmental toxicants as well as lifestyle factors have a crucial role to play. Bisphenol S (BPS) was believed to be a "safer" replacement of bisphenol A (BPA) but recent data documented its neurotoxic, hepatotoxic, nephrotoxic, and reprotoxic attributes. Hence based on the scarcity of reports, we investigated molecular insights into BPS‐induced ovarian dysfunction and protective actions of melatonin against it in adult golden hamsters, Mesocricetus auratus. Hamsters were administered with melatonin (3 mg/kg BW i.p. alternate days) and BPS (150 mg/kg BW orally every day) for 28 days. BPS treatment disrupted hypothalamo–pituitary–ovarian (HPO) axis as evident by reduced gonadotropins such as luteinizing hormone (LH) and follicle‐stimulating hormone (FSH), ovarian steroids such as estradiol (E2) and progesterone (P4), thyroid hormones namely triiodothyronine (T3) and thyroxine (T4) and melatonin levels along with their respective receptors (ERα, TRα, and MT‐1) thereby reducing ovarian folliculogenesis. BPS exposure also led to ovarian oxidative stress/inflammation by increasing reactive oxygen species and metabolic disturbances. However, melatonin supplementation to BPS restored ovarian folliculogenesis/steroidogenesis as indicated by increased number of growing follicles/corpora lutea and E2/P4 levels. Further, melatonin also stimulated key redox/survival markers such as silent information regulator of transcript‐1 (SIRT‐1), forkhead box O‐1 (FOXO‐1), nuclear factor E2‐related factor‐2 (Nrf2), and phosphoinositide 3‐kinase/protein kinase B (PI3K/pAkt) expressions along with enhanced ovarian antioxidant capacity. Moreover, melatonin treatment reduced inflammatory load including ovarian nuclear factor kappa‐B (NFĸB), cyclooxygenase‐2 (COX‐2), and inducible nitric oxide synthase (iNOS) expressions, serum tumor necrosis factor α (TNFα), C‐reactive protein (CRP) and nitrite–nitrate levels as well as upregulated ovarian insulin receptor (IR), glucose uptake transporter‐4 (GLUT‐4), connexin‐43, and proliferating cell nuclear antigen (PCNA) expressions in ovary thereby ameliorating inflammatory and metabolic alterations due to BPS. In conclusion, we found severe deleterious impact of BPS on ovary while melatonin treatment protected ovarian physiology from these detrimental changes suggesting it to be a potential preemptive candidate against environmental toxicant‐compromised female reproductive health. [ABSTRACT FROM AUTHOR]

Although it was described previously for estrogen (E2) regulation of intestinal epithelial Cl− and HCO3− secretion in sex difference, almost nothing is known about the roles of estrogen receptor (ER) subtypes in regulating E2-modulated epithelial ion transports and epithelial restitution. Here, we aimed to investigate ERα and ERβ subtypes in the regulation of E2-modulated colonic epithelial HCO3− and Cl− secretion and epithelial restitution. Through physiological and biochemical studies, in combination of genetic knockdown, we showed that ERα attenuated female colonic Cl− secretion but promoted Ca2+-dependent HCO3− secretion via store-operated calcium entry (SOCE) mechanism in mice. However, ERβ attenuated HCO3 − secretion by inhibiting Ca2+ via the SOCE and inhibiting cAMP via protein kinases. Moreover, ERα but not ERβ promoted epithelial cell restitution via SOCE/Ca2+ signaling. ERα also enhanced cyclin D1, proliferating cell nuclear antigen, and β-catenin expression in normal human colonic epithelial cells. All ERα-mediated biological effects could be attenuated by its selective antagonist and genetic knockdown. Finally, both ERα and ERβ were expressed in human colonic epithelial cells and mouse colonic tissues. We therefore conclude that E2 modulates complex colonic epithelial HCO3− and Cl− secretion via ER subtype-dependent mechanisms and that ERα is specifically responsible for colonic epithelial regeneration. This study provides novel insights into the molecular mechanisms of how ERα and ERβ subtypes orchestrate functional homeostasis of normal colonic epithelial cells. [ABSTRACT FROM AUTHOR]

Proliferating cell nuclear antigen (PCNA) belongs to the DNA sliding clamp family. Via interacting with various partner proteins, PCNA plays critical roles in DNA replication, DNA repair, chromatin assembly, epigenetic inheritance, chromatin remodeling, and many other fundamental biological processes. Although PCNA and PCNA-interacting partner networks are conserved across species, PCNA of a given species is rarely functional in heterologous systems, emphasizing the importance of more representative PCNA studies. Here, we report two crystal structures of PCNA from African swine fever virus (ASFV), which is the only member of the Asfarviridae family. Compared to the eukaryotic and archaeal PCNAs and the sliding clamp structural homologs from other viruses, AsfvPCNA possesses unique sequences and/or conformations at several regions, such as the J-loop, interdomain-connecting loop (IDCL), P-loop, and C-tail, which are involved in partner recognition or modification of sliding clamps. In addition to double-stranded DNA binding, we also demonstrate that AsfvPCNA can modestly enhance the ligation activity of the AsfvLIG protein. The unique structural features of AsfvPCNA can serve as a potential target for the development of ASFV-specific inhibitors and help combat the deadly virus. [ABSTRACT FROM AUTHOR]

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BACKGROUND: MiR-338-3p could inhibit osteoclast differentiation, and downregulation of receptor activator of nuclear factor-κB ligand level could promote bone formation. However, it is unclear whether miR-338-3p can affect the proliferation and apoptosis of alveolar bone osteoblasts by regulating the receptor activator of nuclear factor-κB ligand level. OBJECTIVE: To explore the effect and mechanism of miR-338-3p on proliferation and apoptosis of alveolar bone osteoblasts by targeting receptor activator of nuclear factor-κB ligand. METHODS: Human alveolar bone osteoblasts were isolated, transfected and treated with Wnt-C59 (Wnt/β-catenin pathway inhibitor), and divided into transfection control group, miR-338-3p group, miR-338-3p+control group, miR-338-3p+receptor activator of nuclear factor-κB ligand group and miR-338-3p+ Wnt-C59 group. The dual luciferase report experiment was used to verify the regulatory effect of miR-338-3p on receptor activator of nuclear factor-κB ligand. Cell counting kit-8 and 5-Ethynyl-2’-deoxyuridine staining were used to detect cell proliferation levels. Flow cytometry was used to detect cell cycle and apoptosis levels. RT-qPCR was used to detect miR-338-3p, receptor activator of nuclear factor-κB ligand, Wnt-3a, β-Catenin, glycogen synthase kinase-3β mRNA levels. Western blot was used to detect RANKL, proliferating cell nuclear antigen, Ki67, CyclinD1, B-cell lymphoma/leukemia-2, B-cell lymphoma-2 related X protein, Caspase3, Wnt-3a, β-catenin, glycogen synthase kinase-3β protein levels. RESULTS AND CONCLUSION: miR-338-3p could target the regulation of receptor activator of nuclear factor-κB ligand. After overexpression of miR-338-3p, cell survival rate, 5-Ethynyl-2’-deoxyuridine positive cell rate, proportion of S-phase cells were increased, and apoptosis rate was decreased. The mRNA and protein levels of miR-338-3p, proliferating cell nuclear antigen, Ki67, CyclinD1, B-cell lymphoma/leukemia-2, Wnt-3a, and β-catenin were increased, while the mRNA and protein levels of B-cell lymphoma-2 related X protein, Caspase3 protein, receptor activator of nuclear factor-κB ligand, and glycogen synthase kinase-3β were decreased (all P < 0.05). Overexpression of receptor activator of nuclear factor-κB ligand or Wnt-C59 could weaken the effects of overexpression of miR- 338-3p on cell proliferation and apoptosis (all P < 0.05). Overall, miR-338-3p promotes alveolar bone osteoblast proliferation and inhibits apoptosis by targeting receptor activator of nuclear factor-κB ligand, which may act through activation of the Wnt/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Mismatch repair (MMR) safeguards genome stability through recognition and excision of DNA replication errors.1-4 How eukaryotic MMR targets the newly replicated strand in vivo has not been established. MMR reactions reconstituted in vitro are directed to the strand containing a preexisting nick or gap,5-8 suggesting that strand discontinuities could act as discrimination signals. Another candidate is the proliferating cell nuclear antigen (PCNA) that is loaded at replication forks and is required for the activation of Mlh1-Pms1 endonuclease.7-9 Here, we discovered that overexpression of DNA ligase I (Cdc9) in Saccharomyces cerevisiae causes elevated mutation rates and increased chromatin-bound PCNA levels and accumulation of Pms1 foci that are MMR intermediates, suggesting that premature ligation of replication-associated nicks interferes with MMR. We showed that yeast Pms1 expression is mainly restricted to S phase, in agreement with the temporal coupling between MMR and DNA replication.10 Restricting Pms1 expression to the G2/M phase caused a mutator phenotype that was exacerbated in the absence of the exonuclease Exo1. This mutator phenotype was largely suppressed by increasing the lifetime of replication-associated DNA nicks, either by reducing or delaying Cdc9 ligase activity in vivo. Therefore, Cdc9 dictates a window of time for MMR determined by transient DNA nicks that direct the Mlh1-Pms1 in a strand-specific manner. Because DNA nicks occur on both newly synthesized leading and lagging strands,11 these results establish a general mechanism for targeting MMR to the newly synthesized DNA, thus preventing the accumulation of mutations that underlie the development of human cancer.

Toxoplasma gondii is an intracellular protozoan parasite that has the remarkable ability to infect and replicate in neutrophils, immune cells with an arsenal of antimicrobial effector mechanisms. We report that T. gondii infection extends the life span of primary human peripheral blood neutrophils by delaying spontaneous apoptosis, serum starvation-induced apoptosis, and tumor necrosis alpha (TNF-α)-mediated apoptosis. T. gondii blockade of apoptosis was associated with an inhibition of processing and activation of the apoptotic caspases caspase-8 and -3, decreased phosphatidylserine exposure on the plasma membrane, and reduced cell death. We performed a global transcriptome analysis of T. gondii-infected peripheral blood neutrophils using RNA sequencing (RNA-Seq) and identified gene expression changes associated with DNA replication and DNA repair pathways, which in mature neutrophils are indicative of changes in regulators of cell survival. Consistent with the RNA-Seq data, T. gondii infection upregulated transcript and protein expression of PCNA, which is found in the cytosol of human neutrophils, where it functions as a key inhibitor of apoptotic pro-caspases. Infection of neutrophils resulted in increased interaction of PCNA with pro-caspase-3. Inhibition of this interaction with an AlkB homologue 2 PCNA-interacting motif (APIM) peptide reversed the infection-induced delay in cell death. Taken together, these findings indicate a novel strategy by which T. gondii manipulates cell life span in primary human neutrophils, which may allow the parasite to maintain an intracellular replicative niche and avoid immune clearance.IMPORTANCEToxoplasma gondii is an obligate intracellular parasite that can cause life-threatening disease in immunocompromised individuals and in the developing fetus. Interestingly, T. gondii has evolved strategies to successfully manipulate the host immune system to establish a productive infection and evade host defense mechanisms. Although it is well documented that neutrophils are mobilized during acute T. gondii infection and infiltrate the site of infection, these cells can also be actively infected by T. gondii and serve as a replicative niche for the parasite. However, there is a limited understanding of the molecular processes occurring within T. gondii-infected neutrophils. This study reveals that T. gondii extends the life span of human neutrophils by inducing the expression of PCNA, which prevents activation of apoptotic caspases, thus delaying apoptosis. This strategy may allow the parasite to preserve its replicative intracellular niche.

Abstract Background Osteosarcoma is a type of bone cancer that predominantly affects young individuals, including children and adolescents. The disease progresses through heterogeneous genetic alterations, and patients often develop pulmonary metastases even after the primary tumors have been surgically removed. Ubiquitin-specific peptidases (USPs) regulate several critical cellular processes, such as cell cycle progression, transcriptional activation, and signal transduction. Various studies have revealed the significance of USP37 in the regulation of replication stress and oncogenesis. Methods In this study, the Cancer Genome Atlas (TCGA) database was analyzed to investigate USP37 expression. RNA sequencing was utilized to assess the impact of USP37 overexpression and depletion on gene expression in osteosarcoma cells. Various molecular assays, including colony formation, immunofluorescence, immunoprecipitation, and DNA replication restart, were employed to examine the physical interaction between USP37 and PCNA, as well as its physiological effects in osteosarcoma cells. Additionally, molecular docking studies were conducted to gain insight into the nature of the interaction between USP37 and PCNA. Furthermore, immunohistochemistry was performed on archived tissue blocks from osteosarcoma patients to establish a correlation between USP37 and PCNA expression. Results Analysis of the TCGA database revealed that increased expression of USP37 was linked to decreased progression-free survival (PFS) in osteosarcoma patients. Next-generation sequencing analysis of osteosarcoma cells demonstrated that overexpression or knockdown of USP37 led to the expression of different sets of genes. USP37 overexpression provided a survival advantage, while its depletion heightened sensitivity to replication stress in osteosarcoma cells. USP37 was found to physically interact with PCNA, and molecular docking studies indicated that the interaction occurs through unique residues. In response to genotoxic stress, cells that overexpressed USP37 resolved DNA damage foci more quickly than control cells or cells in which USP37 was depleted. The expression of USP37 varied in archived osteosarcoma tissues, with intermediate expression seen in 52% of cases in the cohort examined. Conclusion The results of this investigation propose that USP37 plays a vital role in promoting replication stress tolerance in osteosarcoma cells. The interaction between USP37 and PCNA is involved in the regulation of replication stress, and disrupting it could potentially trigger synthetic lethality in osteosarcoma. This study has expanded our knowledge of the mechanism through which USP37 regulates replication stress, and its potential as a therapeutic target in osteosarcoma merits additional exploration.

Eukaryotic DNA Mismatch Repair (MMR) involves redundant exonuclease 1 (Exo1)-dependent and Exo1-independent pathways, of which the Exo1-independent pathway(s) is not well understood. The exo1Δ440-702 mutation, which deletes the MutS Homolog 2 (Msh2) and MutL Homolog 1 (Mlh1) interacting peptides (SHIP and MIP boxes, respectively), eliminates the Exo1 MMR functions but is not lethal in combination with rad27Δ mutations. Analyzing the effect of different combinations of the exo1Δ440-702 mutation, a rad27Δ mutation and the pms1-A99V mutation, which inactivates an Exo1-independent MMR pathway, demonstrated that each of these mutations inactivates a different MMR pathway. Furthermore, it was possible to reconstitute a Rad27- and Msh2-Msh6-dependent MMR reaction in vitro using a mispaired DNA substrate and other MMR proteins. Our results demonstrate Rad27 defines an Exo1-independent eukaryotic MMR pathway that is redundant with at least two other MMR pathways.

ABSTRACT Purpose: We aimed to investigate the antioxidant activity of nebivolol against possible damage to the ovarian tissue due to the application of deltamethrin as a toxic agent, by evaluating histopathological proliferating cell nuclear antigen (PCNA) and tumor necrosis factor-alpha (TNF-α) signal molecules immunohistochemically. Methods: The animals were divided into three groups as control, deltamethrin and deltamethrin + nebivolol groups. Vaginal smears were taken after the animals were mated and detected on the first day of pregnancy. After the sixth day, deltamethrin (0.5 mL of 30 mg/kg BW undiluted ULV), and 2 mL of sterile nebivolol solution were administered intraperitoneally every day for 6-21 periods. After routine histopathological follow-up, the ovarian tissue was stained with hematoxylin and eosin stain. Results: Control group showed normal histology of ovarium. In deltamethrin group, hyperplasic cells, degenerative follicles, pyknotic nuclei, inflammation and hemorrhagic areas were observed. Nebivolol treatment restored these pathologies. Deltamethrin treatment increased TNF-α and PCNA reaction. However, nebivolol decreased the expression. Conclusions: It was thought that deltamethrin toxicity adversely affected follicle development by inducing degeneration and apoptotic process in preantral and antra follicle cells, and nebivolol administration might reduce inflammation and slow down the apoptotic signal in the nuclear phase and regulate reorganization.

ABSTRACT: This study investigated the detrimental effects of diethylstilbestrol (DES), an estrogenic endocrine-disrupting chemical, on the viability of primordial germ cells (PGCs), embryonic precursors of germ cells, in Japanese quail. We injected 50 or 100 nmol DES solubilized in sesame oil into the yolk of stage X embryos and assessed changes in the population and cell cycle properties of circulating PGCs in blood vessels and gonadal PGCs after 2.5- and 7-day incubations, respectively. Liquid chromatography tandem mass spectrometer and Western blotting analyses identified DEAD-box polypeptide 4 (DDX4) and proliferating cell nuclear antigen (PCNA) as a stem cell marker and proliferation marker of quail PGCs, respectively. Immunochemical analyses revealed significant decreases in the number of DDX4- and PCNA-positive blood-circulating PGCs in males treated with 50 and 100 nmol DES than in the oil-treated control group. These reductions were not observed in females. Furthermore, the number of DDX4-positive gonadal PGCs was smaller in males treated with 50 and 100 nmol DES than in the control group, and these reductions were not observed in females. The protein expression of the Sertoli cell marker showed normal testis development in DES-treated embryos on d 7. These results demonstrate the potentially cytotoxic effects of DES on male germ cells, namely, the inhibition of cell cycle progression and induction of apoptosis in Japanese quail.

Proliferating cell nuclear antigen (PCNA) forms a homotrimer that encircles replicating DNA and is bound by DNA polymerases to add processivity to cellular DNA synthesis. In addition, PCNA acts as a scaffold to recruit DNA repair and chromatin remodeling proteins to replicating DNA via its interdomain connecting loop (IDCL). Despite encoding a DNA polymerase processivity factor UL42, it was previously found that PCNA associates with herpes simplex virus type 1 (HSV-1) replication forks and is necessary for productive HSV-1 infection. To define the role that PCNA plays during viral DNA replication or a replication-coupled process, we investigated the effects that two mechanistically distinct PCNA inhibitors, PCNA-I1 and T2AA, have on the HSV-1 infectious cycle. PCNA-I1 binds at the interface between PCNA monomers, stabilizes the homotrimer, and may interfere with protein-protein interactions. T2AA inhibits select protein-protein interactions within the PCNA IDCL. Here we demonstrate that PCNA-I1 treatment results in reduced HSV-1 DNA replication, late gene expression, and virus production, while T2AA treatment results in reduced late viral gene expression and infectious virus production. To pinpoint the mechanisms by which PCNA inhibitors affect viral processes and protein recruitment to replicated viral DNA, we performed accelerated native isolation of proteins on nascent DNA (aniPOND). Results indicate that T2AA inhibits recruitment of the viral uracil glycosylase UL2 and transcription regulatory factors to viral DNA, likely leading to a defect in viral base excision repair and the observed defect in late viral gene expression and infectious virus production. In addition, PCNA-I1 treatment results in decreased association of the viral DNA polymerase UL30 and known PCNA-interacting proteins with viral DNA, consistent with the observed block in viral DNA replication and subsequent processes. Together, we conclude that inhibitors of cellular PCNA block recruitment of key viral and cellular factors to viral DNA to inhibit viral DNA synthesis and coupled processes. Author summary: The goal of this study was to determine what role PCNA associated with HSV-1 replication forks plays during viral infection. Using two commercially available PCNA inhibitors, we provide evidence that targeting specific functions of PCNA block HSV-1 DNA replication and subsequent steps in the infectious cycle. Although it was previously thought that viral factors alone facilitate the process of HSV-1 DNA replication, results presented here contradict this long-standing view. With the continued emergence of antiviral resistant HSV-1 variants, these results provide insight into how cellular proteins can be targeted for antiviral treatment. [ABSTRACT FROM AUTHOR]

Background: Protein biomarkers of cancer progression and response to therapy are increasingly important for improving personalized medicine. Advanced quantitative pathology platforms enable measurement of protein expression in tissues at the single-cell level. However, this rich quantitative cell-by-cell biomarker information is most often not exploited. Instead, it is reduced to a single mean across the cells of interest or converted into a simple proportion of binary biomarker-positive or -negative cells. Results: We investigated the utility of retaining all quantitative information at the single-cell level by considering the values of the quantile function (inverse of the cumulative distribution function) estimated from a sample of cell signal intensity levels in a tumor tissue. An algorithm was developed for selecting optimal cutoffs for dichotomizing cell signal intensity distribution quantiles as predictors of continuous, categorical or survival outcomes. The proposed algorithm was used to select optimal quantile biomarkers of breast cancer progression based on cancer cells' cell signal intensity levels of nuclear protein Ki-67, Proliferating cell nuclear antigen, Programmed cell death 1 ligand 2, and Progesterone receptor. The performance of the resulting optimal quantile biomarkers was validated and compared to the standard cancer compartment mean signal intensity markers using an independent external validation cohort. For Ki-67, the optimal quantile biomarker was also compared to established biomarkers based on percentages of Ki67-positive cells. For proteins significantly associated with PFS in the external validation cohort, the optimal quantile biomarkers yielded either larger or similar effect size (hazard ratio for progression-free survival) as compared to cancer compartment mean signal intensity biomarkers. Conclusion: The optimal quantile protein biomarkers yield generally improved prognostic value as compared to the standard protein expression markers. The proposed methodology has a broad application to single-cell data from genomics, transcriptomics, proteomics, or metabolomics studies at the single cell level. [ABSTRACT FROM AUTHOR]

Sustainable farming of fish species depends on emerging new feed ingredients, which can alter the features of the digestive tract and influence animals' overall health. Recent research has shown that functional feeds hold great potential for enhancing fish robustness by evoking appropriate responses at the intestine level. However, there is a lack of extensive and accurate descriptions of the morphology of the gastrointestinal tract of most farmed fish. We have characterised the intestine of European seabass thoroughly, by targeting four segments − anterior, mid, posterior and rectum. Results indicated that the anterior segment is mostly associated with absorption-related features; this segment has the largest absorptive area, the longest villi, and the highest number of neutral goblet cells (GC). The posterior segment and rectum have distinct histomorphometric features, but both seem to be important for immunity, displaying the highest count of acid GC and the highest expression of immune-related genes. The strongest proliferating cell nuclear antigen (PCNA) signal was observed in the anterior intestine and rectum, with PCNA+ cells appearing at the base of the villi and the corresponding villi branches. We have also evaluated the impact of a novel feed supplemented with a macro- and microalgae blend and found that there were no differences in terms of growth. However, the alterations observed in the mid intestine of fish fed the blend, such as thickening of the submucosa and lamina propria, an increased number of leucocytes, and higher expression of immune- and oxidative stress-related genes, suggest that algae may have an immunomodulatory effect. In the current article, we have described the morphology and expression patterns of the intestine segments of European seabass in detail and have presented a comprehensive report of the indices and methods used for the semi-quantitative and quantitative histomorphometric assessments, thereby providing useful information for future studies that aim to maintain intestinal health through dietary interventions. [ABSTRACT FROM AUTHOR]

BACKGROUND: Acute spinal cord injury is a disease with high disability and mortality. An active and effective treatment in the acute phase is of great significance to prevent secondary injury. OBJECTIVE: To investigate the mechanism underlying Tongfu Zhuyu therapy in the treatment of acute spinal cord injury through observing the therapeutic effect of Jiajian Didang Tang on edema and recovery of limb function after acute spinal cord injury in rats. METHODS: A total of 60 Sprague-Dawley rats were randomly divided into sham group, model group, Didang Tang group, 2-(nicotinamide)-1,3,4-thiadiazole (TGN-020) group, and Didang Tang combined with TGN-020 group (combined group), with 12 rats in each group. Among them, the sham group only underwent laminectomy without spinal cord shock and the other groups used NYU percussion device to create an acute spinal cord injury model. After modeling, the sham and model groups did not take any treatment measures. The Didang Tang group was given 5.04 g/kg Didang Tang by intragastric administration, the TGN020 group was given 5 mg/kg TGN-020 by intraperitoneal injection, and the combined group was given TGN-020 by intraperitoneal injection combined with Didang Tang by intragastric administration, once a day for 7 days. Basso, Beattie and Bresnahan score and Reuter score were evaluated on 1, 3, 5 and 7 days after operation. Spinal cord tissue was taken for general observation and water content measurement at 7 days after operation. The degree of tissue damage was detected by hematoxylin-eosin staining. The number of aquaporin 4/glial fibrillary acidic protein-positive cells in the spinal cord tissue was detected by immunofluorescence assay. The protein expression levels of Aquaporin 4, glial fibrillary acidic protein, chondroitin sulfate proteoglycan, and proliferating cell nuclear antigen protein in spinal cord tissue were detected by western blot. RESULTS AND CONCLUSION: The behavioral scores showed that compared with the model group, the Basso, Beattie and Bresnahan scores were significantly increased in the three administration groups (P < 0.05), while the Reuter scores were significantly decreased (P < 0.05). Moreover, there was no significant difference among the three administration groups (P > 0.05). Compared with the model group, the water content of spinal cord tissue was significantly decreased in the three administration groups (P < 0.05), while there was no significant difference among the three administration groups (P > 0.05). Hematoxylin-eosin staining results showed that the structure of the model group was disorganized, with a lot of scar tissue and syringomyelia. Compared with the model group, the tissue structure was relatively complete in the three administration group, with a small amount of scar tissue hyperplasia and syringomyelia, and the tissue morphology was effectively improved. Immunofluorescence staining results showed that compared with the model group, the number of aquaporin 4/glial fibrillary acidic protein-positive cells was less in the three administration groups, with lower brightness and reduced degree of cell swelling (P < 0.05). However, there was no significant difference among the three administration groups (P > 0.05). Western blot results showed that compared with the model group, the expressions of aquaporin 4, glial fibrillary acidic protein, chondroitin sulfate proteoglycan, and proliferating cell nuclear antigen protein in the spinal cord tissue of the three administration groups were significantly decreased (P < 0.05). While, there was no significant difference among the three administration groups (P > 0.05). To conclude, Jiajian Didang Tang can promote the recovery of limb function in rats with acute spinal cord injury under the guidance of Tongfu Zhuyu therapy. Its mechanism may reduce edema at the injury site by inhibiting the expression of aquaporin 4, so as to effectively inhibit the activation and proliferation of reactive astrocytes and reduce the formation of glial scar. [ABSTRACT FROM AUTHOR]

Proliferating cell nuclear antigen (PCNA) is the key regulator of human DNA metabolism. One important interaction partner is p15, involved in DNA replication and repair. Targeting the PCNA–p15 interaction is a promising therapeutic strategy against cancer. Here, a Förster resonance energy transfer (FRET)-based assay for the analysis of the PCNA–p15 interaction was developed. Next to the application as screening tool for the identification and characterization of PCNA–p15 interaction inhibitors, the assay is also suitable for the investigation of mutation-induced changes in their affinity. This is particularly useful for analyzing disease associated PCNA or p15 variants at the molecular level. Recently, the PCNA variant C148S has been associated with Ataxia-telangiectasia-like disorder type 2 (ATLD2). ATLD2 is a neurodegenerative disease based on defects in DNA repair due to an impaired PCNA. Incubation time dependent FRET measurements indicated no effect on PCNAC148S–p15 affinity, but on PCNA stability. The impaired stability and increased aggregation behavior of PCNAC148S was confirmed by intrinsic tryptophan fluorescence, differential scanning fluorimetry (DSF) and asymmetrical flow field-flow fractionation (AF4) measurements. The analysis of the disease associated PCNA variant demonstrated the versatility of the interaction assay as developed. [ABSTRACT FROM AUTHOR]

Endometriosis is a chronic disease characterized by pelvic inflammation. This study aimed at investigating the molecular mechanisms underlying the pathology and how they can be modulated by the administration of a natural compound, Actaea racemosa (AR). We employed an in vivo model of endometriosis in which rats were intraperitoneally injected with uterine fragments from donor animals. During the experiment, rats were monitored by abdominal high-frequency ultrasound analysis. AR was able to reduce the lesion's size and histological morphology. From a molecular point of view, AR reduced hyperproliferation, as shown by Ki-67 and PCNA expression and MAPK phosphorylation. The impaired apoptosis pathway was also restored, as shown by the TUNEL assay and RT-PCR for Bax, Bcl-2, and Caspase levels. AR also has important antioxidant (reduced Nox expression, restored SOD activity and GSH levels, and reduced MPO activity and MDA levels) and anti-inflammatory (reduced cytokine levels) properties. Moreover, AR demonstrated its ability to reduce the pain-like behaviors associated with the pathology, the neuro-sensitizing mediators (c-FOS and NGF) expression, and the related central astrogliosis (GFAP expression in the spinal cord, brain cortex, and hippocampus). Overall, our data showed that AR was able to manage several pathways involved in endometriosis suppression. [ABSTRACT FROM AUTHOR]

The Wnt/β-catenin signaling pathway plays important roles in the multi-phases of wound healing: homeostasis, inflammation, proliferative, and remodeling phases. However, there are no clinically available therapeutic agents targeting the Wnt/β-catenin pathway. In this study, we tested the effect of 5, 6-dichloroindirubin-3′-methoxime (KY19382), a small molecule that activates the Wnt/β-catenin pathway via interference with the function of the negative feedback regulator CXXC5, on cutaneous wound healing. KY19382 significantly enhanced cell migration of human keratinocytes and dermal fibroblasts with increased levels of β-catenin, phalloidin, Keratin 14, proliferating cell nuclear antigen (PCNA), Collagen I, and alpha-smooth muscle actin (α-SMA) by activating the Wnt/β-catenin signaling pathway without causing significant cytotoxicity. In addition, levels of Collagen I, Keratin 14, PCNA, and stem cell markers were significantly increased by KY19382 in a cutaneous murine wound healing model. Moreover, KY19382 treatment accelerated re-epithelialization and neo-epidermis formation with collagen deposition and stem cell activation at an early stage of cutaneous wound healing. Overall, KY19382 accelerates wound healing via activating the Wnt/β-catenin pathway, and may have the potential to be used for the development of a new wound healing agent. [ABSTRACT FROM AUTHOR]

Triple-negative breast cancer (TNBC) is a heterogeneous breast cancer subtype with poor prognoses and limited therapeutic options. The TATA-box binding protein associated factor 1 (TAF1) is an essential protein involved in the transcriptional regulation of cancer development and progress. However, the therapeutic potential and underlying mechanism of targeting TAF1 in TNBC remain unknown. Here, using chemical probe BAY-299, we identify that TAF1 inhibition leads to the induction of endogenous retrovirus (ERVs) expression and double-stranded RNA (dsRNA) formation, resulting in the activation of interferon responses and cell growth suppression in a subset of TNBC, resembling anti-viral mimicry effect. This correlation between TAF1 and interferon signature was validated in three independent breast cancer patient datasets. Furthermore, we observe heterogeneous responses to TAF1 inhibition across a set of TNBC cell lines. By integrating transcriptome and proteome data, we demonstrate that high levels of proliferating cell nuclear antigen (PCNA) protein serve as a predictive biomarker associated with suppressive tumor immune responses in various cancers, which may limit the efficiency of TAF1 inhibition. • Identification of TAF1 as a novel pharmacological target for triple-negative breast cancer (TNBC). • Discovery of the negative correlation between TAF1 expression and tumor immunity in TNBC. • Detailed elucidation of the molecular mechanism underlying interferon activation upon TAF1 inhibition. • Identification of high PCNA level as a predictive biomarker for suppressive immunity limiting TAF1 inhibition efficacy. [ABSTRACT FROM AUTHOR]

Antinuclear antibody detection by indirect immunofluorescence (IIF) is considered a hallmark for the diagnosis of systemic autoimmune rheumatic diseases. Proliferating cell nuclear antigen (PCNA) pattern is being increasingly detected in infectious diseases. Infections can mimic features of autoimmune disease and may trigger autoimmunity in genetically predisposed individuals. These may lead to a diagnostic dilemma in certain situations. A middle-aged woman, with comorbidities of diabetes mellitus and systemic hypertension, presented with a history of polyarthralgia, fever, altered sensorium, and status epilepticus. Examination showed hepatosplenomegaly and axillary lymphadenopathy. Investigations revealed elevated acute phase reactants, bicytopenia, transaminitis, and wedge infarct in spleen with coexistence of urosepsis with Escherichia coli and tropical infection. Immunological tests detected PCNA by IIF and immunoblot. The patient was treated with sensitive parenteral antibiotics and on improvement discharged. Possible explanation could be due to synthesis of biofilm by invading bacteria would have resulted in formation of an amyloid scaffold such as curli. This could have transported and presented antigens for autoantibody production. Diabetes mellitus, a state of chronic inflammation, would have accentuated NETosis. Differentiating systemic lupus erythematosus from infectious diseases is challenging. They share similar pathogenesis and clinical features. The need for the initiation of immunosuppressive therapy in lupus necessitates for absolute need for new biomarkers and tests for its differentiation from infectious diseases. [ABSTRACT FROM AUTHOR]

Eukaryotic translation initiation factors (eIFs) are genes related to tumor formation. They selectively translate some mRNAs, regulate protein synthesis, promote cell proliferation, and effectively control the progression of some tumors. As a member of this family, eukaryotic translation initiation factor 6 (eIF6) plays a crucial role in tumor formation and progression. Exploring the expression characteristics of eIF6 and proliferating cell nuclear antigen (PCNA), a proliferation-associated factor, in colorectal adenocarcinoma (CRAC) and analyzing their correlation and clinical significance can provide a theoretical basis for the study of CRAC and objective biological indicators for the clinical judgment of tumor formation and progression. The GEPIA database was used to predict the trend of eIF6 and PCNA in CRAC. Sixty-four patients diagnosed with CRAC and undergoing surgical treatment were selected from January 2017 to November 2022. CRAC tissues and normal mucous tissues (>3 cm away from the tumor margin) were retained. The expression of eIF6 and PCNA was detected by immunohistochemistry (IHC). Colon cancer cell lines SW480, HCT15, and SW620 and normal colon cell line NCM460 were selected, and the expression of eFI6 and PCNA in these cells was detected by Western blot. A trend of elevated expression of eIF6 and PCNA was predicted in CRAC by the GEPIA database. The expression levels of eIF6 (65.63 % vs. 9.38 %, X2=43.2000, p<0.0001) and PCNA (84.38 % vs. 46.88 %, X2=19.9481, p<0.0001) were higher in CRAC tissues than that in normal mucosa as indicated by IHC. Significant difference in eIF6 and PCNA expression was found among different maxim tumor diameters and depths of infiltration in colon adenocarcinoma (p<0.05). No statistical difference in eIF6 and PCNA expression was observed among different genders, ages, degrees of differentiation, LNMets, intravascular cancer thrombosis, TNM stages, and numbers of lymph node metastases (p>0.05). A positive correlation was found between eIF6 and PCNA in CRAC (X2=5.05, r=0.77, p=0.0283). Their expression was significantly higher in SW480, HCT15, and SW620 than in NCM460 (p<0.05). eIF6 is highly expressed in CRAC, participates in tumor formation and progression, and has a positive correlation with PCNA. [ABSTRACT FROM AUTHOR]

Objective: Stanniocalcin-2 (STC2), a secreted glycoprotein that is involved in the regulation of angiogenesis, was proposed as one of the mechanisms of neovascularization in hemangioma (HA). We aimed to investigate the effect of STC2 on proliferation and angiogenesis in hemangioma-derived endothelial cells. Methods: The hemangioma samples from HA patients with the median age of six months were surgically collected in the Affiliated Hospital of Weifang Medical University from October 2019 to June 2021, and divided into normal skin tissues (n=10), involuting-phase HAs (n=10) and proliferating-phase HAs (n=10) according to the Mulliken classification. The expression of STC2 was detected in involuting-phase HAs and proliferating-phase HAs. Hemangioma endothelial cells (HemEC) were transfected with small interfering RNA (siRNA) specific for STC2, and cell survival and tube formation were analyzed. Results: STC2 expression in proliferating-phase HAs was markedly higher than in the normal skin tissues and involvingphase HAs. Similarly, STC2 expression was higher in HemEC compared to the control human umbilical vein endothelial cells (HUVEC). Knockdown of STC2 slowed the proliferation of HemEC and decreased the expression of proliferating cell nuclear antigen (PCNA) in HemEC. Moreover, knockdown of STC2 in HemEC inhibited vascular endothelial cell angiogenesis and regulated the expression and phosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2). Mechanistically, STC2 knockdown attenuated the activation of Akt/eNOS signaling, which was abolished by insulin growth factor-1 (IGF-1), the activator of Akt signaling, accompanying by increased proliferation and tube formation of HemEC. Conclusion: Inhibition of STC2 suppresses HemEC proliferation and angiogenesis by VEGFR2/Akt/eNOS pathway, which warrants further development of STC2-based strategies for HA treatment. [ABSTRACT FROM AUTHOR]

Quercetin is an essential flavonoid mostly found in herbal plants, fruits, and vegetables, which exhibits anti‐hypertension properties. However, its pharmacological impact on angiotensin II (Ang II) induced the increase of blood pressure along with in‐depth mechanism needs further exploration. The present study pointed out the anti‐hypertensive role of quercetin and its comprehensive fundamental mechanisms. Our data showed that quercetin treatment substantially reduced the increase in blood pressure, pulse wave velocity, and aortic thickness of abdominal aorta in Ang II‐infused C57BL/6 mice. RNA sequencing revealed that quercetin treatment reversed 464 differentially expressed transcripts in the abdominal aorta of Ang II‐infused mice. Moreover, overlapping KEGG‐enriched signaling pathways identified multiple common pathways between the comparison of Ang II versus control and Ang II + quercetin versus Ang II. Likewise, these pathways included cell cycle as well as p53 pathways. Transcriptome was further validated by immunohistochemistry, indicating that quercetin treatment significantly decreased the Ang II‐induced expression of proliferating cell nuclear antigen (PCNA), cyclin‐dependent kinase‐4 (CDK4), and cyclin D1, while increased protein expression of p53, and p21 in abdominal aortic tissues of mice. In vitro, quercetin treatment meaningfully decreased the cell viability, arrested cell cycle at G0/G1 phase, and up‐regulated the p53 and p21 proteins expression, as well as down‐regulated the protein expression of cell cycle‐related markers, for example, CDK4, cyclin D1 in Ang II stimulated vascular smooth muscle cells (VSMCs). This study addresses pharmacologic and mechanistic perspectives of quercetin against Ang‐II‐induced vascular injury and the increase of blood pressure. [ABSTRACT FROM AUTHOR]

Methotrexate (MTX) is a chemotherapeutic medicine frequently used to treat various forms of cancer. The objective of our research was to examine whether allicin (ALC) and lycopene (LP) could alleviate the harmful effects of MTXinduced testicular toxicity in rats. A total of 49 male white albino rats were divided into 7 groups (7 rats in each group). The rats of group 1 was kept as a control group and received normal saline orally; while rats of group 2 and group 3 administered ALC (20mg/kg/BW) LP (10mg/kg/BW) orally. On day 15, the rats of group 4 received a single dose of MTX (20mg/kg, IP). In group 5, rats received ALC and MTX; in group 6, rats received LP and MTX, and in group 7, rats received ALC, LP, and MTX. Results showed that MTX induced significant increase in values of testicular malondialdehyde (MDA) levels with a substantial reduction in glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and lowered the serum testosterone, follicle-stimulating hormone (FSH), and lutenizing hormone (LH) levels. MTX induced a marked decline in seminiferous tubule diameter and epithelium height. MTX showed a widening of the interstitial space of seminiferous tubules, atrophy, vacuolar degeneration of the germinal epithelium and Leydig's interstitial cells had scanty cytoplasm and vesicular nuclei. Also, testicular proliferating cell nuclear antigen (PCNA) expression was recorded. In MTX-intoxicated rats, ALC and LP increased antioxidant biomarkers, decreased MDA, and attenuated changes in serum hormonal parameters. It is concluded that ALC and LP have significant protective effects on MTX- induced testicular toxicity due to their antioxidant properties. [ABSTRACT FROM AUTHOR]

Paclitaxel (PTX) is an effective chemotherapeutic agent for cancer patients. It has been reported that circular RNA (circRNA) circ_0005785is involved in the progression of hepatocellular carcinoma (HCC). The purpose of this study is to explore the role and mechanism of circ_0005785 in the PTX resistance of HCC. Cell viability, proliferation, invasion, migration, apoptosis, and angiogenesis were detected using 3‐(4, 5‐dimethyl‐2‐thiazolyl)‐2, 5‐diphenyl‐2‐H‐tetrazolium bromide (MTT), colony formation, transwell, wound‐healing, flow cytometry, and tube formation assay. Circ_0005785, microRNA‐640 (miR‐640), and Glycogen synthase kinase‐3 beta (GSK3β) levels were detected using real‐time quantitative polymerase chain reaction. Protein levels of Proliferating cell nuclear antigen (PCNA), Bcl‐2, and GSK3β were measured using western blot assay. After being predicted using Circular RNA interactome or TargetScan, binding between miR‐640 and circ_0005785 or GSK3β was verified using dual‐luciferase reporter and RNA Immunoprecipitation assay. PTX treatment could repress HCC cell viability, decrease circ_0005785 and GSK3β expression, and increase the miR‐640 level in HCC cell lines. Furthermore, circ_0005785 and GSK3β were increased, and miR‐640 was decreased in HCC tissues and cell lines. Moreover, circ_0005785 knockdown hindered proliferation, migration, invasion, angiogenesis, and boosted apoptosis in PTX‐treated HCC cells in vitro. In addition, circ_0005785 silencing improved the PTX sensitivity of HCC in vivo. Mechanistically, circ_0005785 acted as a sponge of miR‐640 to regulate GSK3β expression. PTX restrained HCC tumorigenesis partly via regulating the circ_0005785/miR‐640/GSK3β axis, hinting at a promising therapeutic target for the HCC treatment. [ABSTRACT FROM AUTHOR]

This article reviews the currently used therapeutic strategies to target DNA replication stress for cancer treatment in the clinic, highlighting their effectiveness and limitations due to toxicity and drug resistance. Cancer cells experience enhanced spontaneous DNA damage due to compromised DNA replication machinery, elevated levels of reactive oxygen species, loss of tumor suppressor genes, and/or constitutive activation of oncogenes. Consequently, these cells are addicted to DNA damage response signaling pathways and repair machinery to maintain genome stability and support survival and proliferation. Chemotherapeutic drugs exploit this genetic instability by inducing additional DNA damage to overwhelm the repair system in cancer cells. However, the clinical use of DNA-damaging agents is limited by their toxicity and drug resistance often arises. To address these issues, the article discusses a potential strategy to target the cancer-associated isoform of proliferating cell nuclear antigen (caPCNA), which plays a central role in the DNA replication and damage response network. Small molecule and peptide agents that specifically target caPCNA can selectively target cancer cells without significant toxicity to normal cells or experimental animals. [ABSTRACT FROM AUTHOR]

The evidence that purified pol2-M644G DNA polymerase (Pol)e exhibits a highly elevated bias for forming T:dTTP mispairs over A:dATP mispairs and that yeast cells harboring this Pole mutation accumulate A > T signature mutations in the leading strand have been used to assign a role for Pole in replicating the leading strand. Here, we determine whether A > T signature mutations result from defects in Pole proofreading activity by analyzing their rate in Pole proofreading defective pol2-4 and pol2-M644G cells. Since purified pol2-4 Pole exhibits no bias for T:dTTP mispair formation, A > T mutations are expected to occur at a much lower rate in pol2-4 than in pol2-M644G cells if Pole replicated the leading strand. Instead, we find that the rate of A > T signature mutations are as highly elevated in pol2-4 cells as in pol2-M644G cells; furthermore, the highly elevated rate of A > T signature mutations is severely curtailed in the absence of PCNA ubiquitination or Pol? in both the pol2-M644G and pol2-4 strains. Altogether, our evidence supports the conclusion that the leading strand A > T signature mutations derive from defects in Pole proofreading activity and not from the role of Pole as a leading strand replicase, and it conforms with the genetic evidence for a major role of Pold in replication of both the DNA strands. [ABSTRACT FROM AUTHOR]

African swine fever virus (ASFV) is an important animal pathogen that is causing a current African swine fever pandemic and affecting pork industry globally. ASFV encodes at least 150 proteins, and the functions of many of them remain to be clarified. The ASFV protein E301R (pE301R) was predicted to be a DNA sliding clamp protein homolog working as a DNA replication processivity factor. However, structural evidence was lacking to support the existence of a ring-shaped sliding clamp in large eukaryotic DNA viruses. Here, we have solved a high-resolution crystal structure of pE301R and identified a canonical ring-shaped clamp comprising a pE301R trimer. Interestingly, this complete-toroidal structure is different from those of the monomeric clamp protein homolog, herpes simplex virus UL42, and the C-shaped dimeric human cytomegalovirus UL44, but highly homologous to that of the eukaryotic clamp homolog proliferating cell nuclear antigen. Moreover, pE301R has a unique N-terminal extension that is important in maintaining the trimeric form of the protein in solution, while specific features in length and surface electrostatic potential of its interdomain connector implies specificity in interactions with binding partners such as the viral DNA polymerase. Thus, our data pave the way for further dissection of the processivity clamp protein structural and functional diversity and ASFV DNA replication mechanisms. [ABSTRACT FROM AUTHOR]

To explore the influence and possible mechanism of extracts from Phellinus igniarius (expanded program on immunization) on oral squamous cell carcinoma cell proliferation, migration and invasion. CAL27 cells were treated with expanded program on immunization or transfected with microRNA-655-3p. Furthermore, transfected CAL27 cells were exposed to 600 µg/ml expanded program on immunization. Proliferation, migration and invasion were assessed using 3-(4,5-dimethylthiazole-2-yl)- 2,5-diphenyltetrazolium bromide and Transwell. Matrix metallopeptidase 2, matrix metallopeptidase 9 and proliferating cell nuclear antigen protein levels were monitored using Western blot. After various doses of expanded program on immunization, cell viability, migration, invasion, proliferating cell nuclear antigen, matrix metallopeptidase 2 and matrix metallopeptidase 9 protein levels were reduced, but microRNA-655-3p content was enhanced in a dose-dependent way. Meanwhile, increased microRNA-655-3p might diminish cell proliferation, migration and invasion. Beyond that, microRNA-655-3p knockdown overturned high-dose expanded program on immunization-mediated tumor development repression. Expanded program on immunization could hinder oral squamous cell carcinoma cell development via modulating microRNA-655-3p. [ABSTRACT FROM AUTHOR]

Introduction. Fractures of the lower jaw are one of the most common injuries in the head and neck. Bone regeneration is a key step in the fracture healing process and plays an important role in restoring jaw function. However, the presence of endogenous intoxication can complicate the healing process and lead to a decreased efficiency of the regenerative process. In this article, we examine the relationship between the regenerative process of bone tissue in mandibular fractures and the level of endogenous intoxication. We investigate the mechanisms of bone regeneration and study the effect of endogenous intoxication on the healing process. Our research will help better understand the healing processes of mandibular fractures and develop more effective treatments.The aim of the study was to establish the effects of chronic endogenous intoxication on both intact bone and the process of reparative osteogenesis in mandibular fractures. Materials and methods. Regularities of morphofunctional transformations in bone tissue were studied during spontaneous healing of mandibular fractures in rats against the background of chronic endogenous intoxication after 30, 60 and 90 days. Simulation of chronic endogenous intoxication was performed by repeated administration of tetrachlormethane and bacterial lipopolysaccharide doses. Determination of medium molecular weight substances, as well as their oligopeptide fraction in blood serum were used as integral laboratory and biochemical indicators of ET development. Concentrations of products of free radical oxidation of lipids were also determined in the blood plasma of rats. Monoclonal antibodies to the proliferating cell nuclear antigen PCNA were used to detect proliferative activity in the regenerate and surrounding bone tissue. The morphometric study was performed in accordance with the principles of systemic quantitative analysis. Results and discussion. The formation of an osteoregenerate at mandibular fracture in rats against the background of chronic endotoxicosis is accompanied by a decreased growth of the volume fraction of bone tissue and stromalvascular component which, according to correlation analysis, is associated with the level of medium molecular weight substances and malonic dialdehyde in blood plasma. There is a strong positive correlation between the volume fraction of the regenerate vessels and the level of malonic dialdehyde (r=0.671-0.692). Negative correlations were detected for the numerical̆ density of osteoblasts and osteocytes (r=-0.622) and positive correlations for osteoclasts (r=0.654-0.873). Conclusions. The study provided a lot of morphological evidence of the effect of chronic endogenous intoxication on both intact bone and the process of reparative osteogenesis after mandibular fractures. [ABSTRACT FROM AUTHOR]

The experiment was designed to evaluate the effect of sodium selenite (Na2SeO3) on the proliferation of bovine mammary gland epithelial cells (BMECs), and to assess the underlying mechanisms. The addition of Na2SeO3 at doses ranging from 18 to 140 nM stimulated BMEC proliferation. Nevertheless, high doses of Na2SeO3 (> 70 nM) did not further stimulate BMEC proliferation compared to 35 nM Na2SeO3. Therefore, the addition of 35 nM Na2SeO3 increased the proportion of epithelial cells undergoing DNA replication, mRNA expression of proliferating cell nuclear antigen (PCNA), Cyclin A2 (CCNA2) and Cyclin D1 (CCND1), as well as protein expression of PCNA and Cyclin A1. Transcription and translation of the B-cell lymphoma 2 (BCL2) gene and the ratio of BCL2 to BCL2-associated X 4 (BAX4) genes were significantly elevated, while mRNA and protein expressions of BAX, caspase 3 (CASP3) and caspase 9 (CASP9) genes were significantly reduced as a result of 35 nM Na2SeO3 supplementation. Additionally, both the Akt and mTOR signalling pathways were activated by 35 nM Na2SeO3. On the other hand, the stimulation of BMEC proliferation, altered expression of proliferative and apoptotic genes and proteins, as well as mTOR signalling pathway activation caused by 35 nM Na2SeO3 addition was suppressed by the Akt inhibitor (AKT-IN-1). Likewise, rapamycin-mediated suppression of mTOR completely reversed the 35 nM Na2SeO3-stimulated BMEC proliferation and alteration of proliferous gene and protein expressions, without affecting mRNA or protein expression of genes related to apoptosis and the Na2SeO3-activated Akt signalling pathway. In conclusion, the results implicated that the proliferation of BMECs was stimulated by 35 nM Na2SeO3 through the Akt-mTOR signalling pathway. [ABSTRACT FROM AUTHOR]

Immunohistochemical staining patterns of markers for neurogenesis staging were compared at the identical stage of cerebellar histogenesis between ferrets (aged 20 days) and mice (aged 10 days). Proliferating cell nuclear antigen (PCNA) immunostaining was observed largely in the granular precursors of the external granular layer (EGL) in both ferrets and mice. PCNA‐immunostaining was also found in brain lipid‐binding protein‐immunopositive cells in the internal granular layer and was more abundant in ferrets than in mice. Paired box 6 immunostaining appeared largely in the EGL granular precursors in mice, whereas it emerged in the migrating/differentiating granular precursors in ferrets. These findings revealed that the types and neurogenesis stages of the EGL granular precursors detected by immunohistochemical markers differed between ferrets and mice. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNAs) have been revealed to harbor open reading frames (ORFs) that can be translated into small peptides. The peptides may participate in the pathogenesis of colorectal cancer (CRC). Herein, we investigated the role of a lncRNA BVES-AS1-encoded peptide in colorectal tumorigenesis. Through bioinformatic analysis, lncRNA BVES-AS1 was predicted to have encoding potential and to be associated with poor prognosis of patients with CRC. In CRC cells, BVES-AS1 was validated to encode a 50-aa-length micro-peptide, named BVES-AS1-201-50aa, through a western blotting method. BVES-AS1-201-50aa enhanced cell viability and promoted the migratory and invasive capacities of HCT116 and SW480 CRC cells in vitro, validated via CCK-8 assay and transwell assay, respectively. Immunofluorescence assay showed that BVES-AS1-201-50aa increased the expression of proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase 9 (MMP9) in CRC cells. We further verified that BVES-AS1-201-50aa targeted and activated the Src/mTOR signaling pathway in CRC cells by co-immunoprecipitation (Co-IP) experiment, qualitative proteomic analysis, and western blotting. Our findings demonstrated that BVES-AS1 could encode a micro-peptide, which promoted CRC cell viability, migration, and invasion in vitro. Our current work broadens the diversity and breadth of lncRNAs in human carcinogenesis. [ABSTRACT FROM AUTHOR]

Reactive oxygen species (ROS) is essential for neutrophil extracellular trap formation (NETosis), and generated either by NADPH oxidases (e.g., during infections) or mitochondria (e.g., sterile injury) in neutrophils. We recently showed that ultraviolet (UV) radiation, a sterile injury-inducing agent, dosedependently induced mitochondrial ROS generation, and increasing levels of ROS shifted the neutrophil death from apoptosis to NETosis. Nevertheless, how ROS executes UV-induced NETosis is unknown. In this study, we first confirmed that UV doses used in our experiments generated mitochondrial ROS, and the inhibition of mitochondrial ROS suppressed NETosis (Mitosox, SYTOX, immunocytochemistry, imaging). Next, we showed that UV irradiation extensively oxidized DNA, by confocal imaging of 8-oxyguanine (8-oxoG) in NETs. Immunofluorescence microscopy further showed that a DNA repair protein, proliferating cell nuclear antigen, was widely distributed throughout the DNA, indicating that the DNA repair machinery was active throughout the genome during UV-induced NETosis. Inhibition of specific steps of base excision repair (BER) pathway showed that steps leading up to DNA nick formation, but not the later steps, suppressed UV-induced NETosis. In summary, this study shows that (i) high levels of mitochondrial ROS produced following UV irradiation induces extensive oxidative DNA damage, and (ii) early steps of the BER pathway leading to DNA nicking results in chromatin decondensation and NETosis. Collectively, these findings reveal how ROS induces NOX-independent NETosis, and also a novel biological mechanism for UV irradiation- and -mitochondrial ROS-mediated NETosis. [ABSTRACT FROM AUTHOR]

The disadvantages and side effects of currently available breast cancer (BC) therapies have compelled researchers to seek new therapeutic strategies. This study was designed to investigate the effect of selenium nanoparticles biosynthesized with carvacrol (SeNPs-CV) on breast cancer (MCF-7) cell lines and to explore possible underlying pathways. Flow cytometry, MTT assays, and various biochemical techniques were used to evaluate the anti-proliferative effects of SeNPs-CV on MCF-7 cells. Cytotoxicity assays showed that treatment with SeNPs-CV could effectively reduce MCF-7 cell proliferation and viability in a dose-dependent manner. However, SeNPs-CV had no cytotoxic effect against Vero cells. Furthermore, SeNPs-CV showed better anticancer activity than metal nanoparticles of selenium evidenced by the lower IC50 obtained in MCF-7 cells (8.3 µg/mL versus 41.6 µg/mL, respectively). Treatment with SeNPs-CV directly targeted Bcl-2, Bax, and caspase-3, leading to the mitochondrial leakage of cytochrome C and subsequent activation of the apoptotic cascade in MCF-7 cells. In addition, MCF-7 cells treated with SeNPs-CV exhibited elevated levels of oxidative stress, as indicated by noticeable rises in 8-OHDG, ROS, NO, and LPO, paralleled by significant exhaustion in GSH levels and antioxidant enzymes activity. In addition, the administration of SeNPs-CV induced the inflammatory mediator IL-1β and downregulated the expression of cell-proliferating nuclear antigen (PCNA) in MCF-7 cells, which plays a critical role in apoptosis. Therefore, the ability of SeNPs-CV to fight BC may be due to its ability to induce oxidative stress, inflammation, and apoptosis in tumor cells. These findings demonstrate the therapeutic potential of Se nanoparticles conjugated with CV, which may provide a novel approach for combination chemotherapy in BC. [ABSTRACT FROM AUTHOR]

Numerous eukaryotic DNA processing enzymes, such as DNA polymerases and ligases, bind the processivity factor PCNA, which acts as a platform to recruit and regulate the binding of enzymes to their DNA substrate. Multiple PCNA‐interacting motifs (PIPs) are present in these enzymes, but their individual structural and functional role has been a matter of debate. Recent cryo‐EM reconstructions of high‐fidelity DNA polymerase Pol δ (Pol δ), translesion synthesis DNA polymerase κ (Pol κ) and Ligase 1 (Lig1) bound to a DNA substrate and PCNA demonstrate that the critical interaction with PCNA involves the internal PIP proximal to the catalytic domain. The ancillary PIPs, located in long disordered regions, are instead invisible in the reconstructions, and appear to function as flexible tethers when the enzymes fall off the DNA. In this review, we discuss the recent structural advancements and propose a functional hierarchy for the PIPs in Pol δ, Pol κ, and Lig1. [ABSTRACT FROM AUTHOR]

Agmatinase (AGMAT) is an enzyme that hydrolyzes agmatine to putrescine and urea. In this study, we explored the functions of AGMAT in colorectal cancer (CRC). By performing gain-of-function and loss-of-function experiments, we investigated the roles of AGMAT in proliferation, cell cycle progression, and apoptosis of CRC cells. We also established a colitis-associated colorectal cancer model by challenging mice with azoxymethane (AOM) and dextran sodium sulfate (DSS), and we subsequently silenced AGMAT expression in mice by adeno-associated virus 9 (AAV9)-mediated delivery of short hairpin RNA (shRNA). In vitro experiments showed that overexpression of AGMAT accelerated the proliferation and inhibited the apoptosis of CRC cells, and AGMAT knockdown exhibited the opposite effects. Interestingly, the oncogenic transcription factor, c-Myc, could bind to the AGMAT promoter and transcriptionally increase AGMAT expression in CRC cells. Additionally, c-Myc and AGMAT were upregulated in the colon of AOM/DSS-treated mice, and AGMAT silencing significantly mitigated colitis in AOM/DSS-treated mice, as evidenced by the increased colon length, attenuated crypt damage, and reduced levels of inflammatory indicators (myeloperoxidase, interleukin-6, tumor necrosis factor-α, inducible nitric oxide synthase, and phosphorylated p65) in colon tissues. Notably, AGMAT silencing decreased both the number and size of tumors, reduced expression of proliferating cell nuclear antigen, and inhibited phosphorylation of protein kinase B (AKT) and extracellular signal-regulated kinase in the colon of AOM/DSS-treated mice. Overall, we determined that AGMAT facilitates tumor progression in CRC. Our findings will be helpful in the search for potential therapeutic targets for CRC. [ABSTRACT FROM AUTHOR]

Background: Scutellaria baicalensis is an important medicinal plant used in China. Several compounds have been extracted from S. baicalensis using modern techniques. Baicalin, an active ingredient of S. baicalensis, is widely used clinically. However, its clinical use is limited owing to poor oral bioavailability and intestinal absorption. Objectives: To evaluate the anti-tumor effect of baicalin magnesium salt (BA-MG, a novel magnesium salt form of baicalin with superior water solubility) on the human hepatocellular carcinoma cell line, HepG2. Materials and Methods: Seven groups were established, including three dosages of BA-MG (200, 250, and 300 µg/mL), baicalin, dimethyl sulfoxide (DMSO) (negative control), magnesium sulfate (negative control), and control groups. Cell proliferation was determined using the cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis were detected by flow cytometry. Cell migration was determined using a cell scratch assay. Protein expression was determined using western blotting. Results: In vitro, BA-MG inhibited proliferation, reduced cell cloning and migration abilities, induced cell cycle arrest, and promoted apoptosis of HepG2 cells (all p < 0.05). The effects of baicalin and BA-MG on proliferation and migration, cell cycle, and apoptosis may be attributed to decreasing the expression of Bcl-2, ROCK-1, proliferating cell nuclear antigen (PCNA), and cyclin E, as well as increasing the expression of Bax and caspase-9. The efficacy of BA-MG is superior to that of baicalin at the same dose. Conclusion: BA-MG showed superior effects on inhibiting cell proliferation and inducing apoptosis in liver cancer cells. [ABSTRACT FROM AUTHOR]

Genetic analyses in Saccharomyces cerevisiae suggest that nucleotide excision repair (NER), homologous recombination (HR), and protease-dependent repair pathways coordinately function to remove DNA-protein crosslinks (DPCs) from the genome. DPCs are genomic cytotoxic lesions generated because of the covalent linkage of proteins with DNA. Although NER and HR processes have been studied in pathogenic Candida albicans, their roles in DPC repair (DPCR) are yet to be explored. Proteases like Wss1 and Tdp1 (tyrosyl-DNA phosphodiesterase-1) are known to be involved in DPCR; however, Tdp1 that selectively removes topoisomerase-DNA complexes is intrinsically absent in C. albicans. Therefore, the mechanism of DPCR might have evolved differently in C. albicans. Herein, we investigated the interplay of three genetic pathways and found that RAD51-WSS1-dependent HR and protease-dependent repair pathways are essential for DPC removal, and their absence caused an increased rate of loss of heterozygosity in C. albicans. RAD1 but not RAD2 of NER is critical for DPCR. In addition, we observed truncation of chromosome #6 in the cells defective in both RAD51 and WSS1 genes. While the protease and DNA-binding activities are essential, a direct interaction of Wss1 with the eukaryotic DNA clamp proliferating cell nuclear antigen is not a requisite for the function of Wss1. DPCR-defective C. albicans cells exhibited filamentous morphology, reduced immune cell evasion, and attenuation in virulence. Thus, we concluded that RAD51-WSS1-dependent DPCR pathways are essential for genome stability and candidiasis development. Since no vaccine against candidiasis is available for human use yet, we propose to explore DPCR-defective attenuated strains (rad51Δwss1ΔΔ and rad2ΔΔrad51ΔΔwss1ΔΔ) for whole-cell vaccine development. [ABSTRACT FROM AUTHOR]

Circular RNAs (circRNAs) function as important regulators in the progression of cancers. The role of circRNA_0048764 (circ_0048764) in the development of breast cancer (BC) remains inconclusive. This work investigates the biological function and molecular mechanism of circ_0048764 in BC. Quantitative real-time PCR (qRT-PCR) was conducted to measure the expression levels of circ_0048764, microRNA-578 (miR-578) and high mobility group AT-hook 2 (HMGA2) mRNA. The viability of BC cells was examined by cell counting kit 8 (CCK-8) assay. Besides, cyclin D1, proliferating cell nuclear antigen (PCNA) and HMGA2 expression levels were detected by western blot. The migrative and invasive capability of BC cells were probed by transwell assay. The relationships between miR-578 and circ_0048764 or HMGA2 3′-UTR were validated by dual-luciferase reporter gene assay. Circ_0048764 was highly expressed in BC tissues and cells, which was significantly associated with tumor size (≥2 cm), lymph node status (positive), and higher TNM stage of BC patients. Circ_0048764 depletion suppressed the proliferative, migrative, and invasive abilities of BC cells, which was rescued by transfection of miR-578 inhibitors. The binding sites were verified between circ_0048764 and miR-578. HMGA2 was identified to be a target of miR-578 in BC cells, and circ_0048764 positively regulated HMGA2 expression in BC cells via repressing miR-578. Circ_0048764 promotes BC cell growth, migration and invasion via absorbing miR-578 and up-regulating HMGA2. [ABSTRACT FROM AUTHOR]

Introduction: Psoriasis is a chronic skin inflammatory disease. Bone marrow mesenchymal stem cells (BM-MSCs) and their derived exosomes are known for their immunomodulatory properties. Aim of the Work: To investigate the effect of locally injected BM-MSCs and their derived exosomes in female albino rats subjected to imiquimod (IMQ)-induced psoriasis-like skin inflammation. Materials and Methods: Fifty albino rats (40 females and 10 males) were used. The female animals were randomly classified into four groups; group I (control group), group II (IMQ group) where the rats received topical IMQ once daily for 5 consecutive days, group III (IMQ + BM-MSCs) where the rats received a dose of 1 million BM-MSCs on the first day only in addition to topical IMQ, and group IV (IMQ + BM-MSCs-derived exosomes) where the rats received purified concentrate of exosomes derived from BM-MSCs in addition to topical IMQ. The 10 male albino rats served as the source of BM-MSCs and their derived exosomes. After 5 days, all animals were sacrificed, and skin specimens were processed for light microscopic studies: H&E and immunohistochemical staining for Proliferating Cell Nuclear Antigen (PCNA) and Y-chromosome identification using Real-time PCR. Morphometric measurements and statistical analysis were done for the mean epidermal thickness, the mean count of PCNA-positive keratinocytes in the epidermis, and the mean area of dermal PCNA-positive reaction. Results: The general observations and microscopic examination of sections obtained from group II rats revealed psoriasis-like skin inflammatory reactions including acanthosis, parakeratosis, and marked inflammatory infiltrate. There was a statistically significant increase in the epidermal thickness and PCNA-positive reactions in group II compared to other groups. Groups III and IV showed significant improvement, however, group III showed almost normal histological structure. Conclusion: BM-MSCs and purified exosomes concentrate were shown to significantly ameliorate psoriasis-like inflammatory changes in the skin of animal models. [ABSTRACT FROM AUTHOR]

Introduction: Potassium bromate (KBrO3) is a strong oxidizing agent and utilized as a flour improver. KBrO3 has Carcinogenic and mutagenic effects, it could generate free radicals and reactive oxygen species (ROS). Potassium bromate is accused as irritating material to tissue surfaces which comes in contact with. Riboflavin has a vital role in cellular function and growth. It possesses a potent antioxidant, anti-inflammatory and immune-modulatory effects. Objective: Estimate the effect of Potassium bromate on gastric fundic mucosa and the possible ameliorating effect of riboflavin in adult male albino rats. Material and Methods: Forty adult male albino rats utilized in the current work. The rats were split to control group, Riboflavin-treated group, Potassium bromate-treated group, Potassium bromate and Riboflavin treated group. At the assigned time, the gastric fundic tissues were obtained for biochemical, histological, histochemical and immunohistochemical studies Results: Potassium bromate treated group exhibited marked deterioration of the fundic mucosa as a wide area of epithelial discontinuity, parietal and chief cells appeared with small darkly stained nuclei. Congested blood vessels, and mononuclear cellular infiltrations were observed. Dilatation of rough endoplasmic reticulum cisternae and confluent secretory granules were detected in chief cells. Statistically, there were a highly significant rise in the mean values of optical density for interleukin 6 (IL6) and mean number of Proliferating cell nuclear antigen (PCNA) positive cells. However, these histological changes were alleviated after riboflavin concomitant administration. Conclusion: KBrO3 has been proved to induce structural changes in the fundic mucosa, and these changes can be ameliorated by concomitant administration of riboflavin. [ABSTRACT FROM AUTHOR]