Frontiers in Bioscience-Landmark

Frontiers in Bioscience-Landmark

Published by IMR Press

Online ISSN: 2768-6698

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Print ISSN: 2768-6701

Disciplines: Biochemistry, Molecular Biology, Cell Biology

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The multilayer structure of the epidermis. The image shows multiple layers of the epidermis, the keratinocytes and melanocytes, and part of the dermis.
Wave-length-dependent ultraviolet (UV) radiation penetrates human skin and activates the production of vitamin D3 from the precursor 7-dehydrocholesterol (7-DHC).
The characteristics of young and aged skin as visible to the naked eye (left side: young; right side: aged). Adapted from [44].
The impact of Vitamin D against intrinsic and extrinsic skin aging and age-associated skin diseases. ECM, extracellular matrix; ROS, reactive oxygen species; UVA, ultraviolet A; UVB, ultraviolet B; TIMP, tissue inhibitor metalloproteinase; MMP, matrix metalloproteinase; NF-κB, nuclear factor kappa B.
Common dermatological diseases among the elderly.
Impact of Vitamin D on Skin Aging, and Age-Related Dermatological Conditions

January 2025

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71 Reads

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2 Citations

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68 reads in the past 30 days

Oxidative and Excitatory Neurotoxic Stresses in CRISPR/Cas9-Induced Kynurenine Aminotransferase Knockout Mice: A Novel Model for Despair-Based Depression and Post-Traumatic Stress Disorder

January 2025

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70 Reads

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1 Citation

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Aims and scope


Frontiers in Bioscience-Landmark (FBL) is an open access journal dedicated to the publication of original articles, reviews, methods and commentaries in all areas of cellular and molecular biology. The journal is interested in manuscripts that describe advances in all aspects of cellular and molecular biology in both eukaryotes and prokaryotes, including studies related to biochemistry, biophysics, physiology, pathology, biotechnology, bioinformatics, and other related fields.

Recent articles


The determination of CD14+ and CD68+ cell subpopulations in blood and follicular fluid. (A) Forward (FSC-A) and side (SSC-A) light scattering, gate cells selected, indicating cells. (B) Singlet signals within all signals of the cells gate in the sample. (C) Histogram distribution of CD14– or CD68–positive cells within singlet signals of the sample. (D) CD163–positive cells within singlets. (E) CD206–positive cells within singlets. (F) Dual staining with antibodies against CD14 or CD68 and CD163 within singlets (quadrants highlight gates reflecting the ratio of cell subpopulations of the sample based on the content of CD14– or CD68–positive cells and CD163). (G) Histogram distribution of CD206–positive cells within CD14+CD163+ or CD68+CD163+. SSCA, strip spectral correlation algorithm; FSC-A, forward and side scatter plot; APC-A, allophycocyanin; FITC-A, fluorescein isothiocyanate; PE-A, phycoerythrin.
An annotated heat map of the correlation between the relative content of monocytes and macrophages in blood and follicular fluid, as well as in vitro fertilisation (IVF) parameters, for groups of patients with different embryo qualities and infertility. Note: (A) Group of female infertility due to tubal factors. (B) Male factor female infertility. (C) Other forms of female infertility. (D) Unspecified female infertility. * p-value ≤ 0.05, ** p-value ≤ 0.01, *** p-value ≤ 0. 001. The color scale on the right indicates the correlation coefficient, with blue representing a negative correlation and red representing a positive correlation.
The monocytes and macrophages in blood and follicular fluid in women with obesity. Note: The assessment of inflammatory activity and the distribution of monocytes and macrophages in blood and follicular fluid has potential for predicting the success of IVF treatments. High-quality embryos are more likely to be obtained from women with higher levels of M2-type monocytes and M2-macrophages in their follicular fluid. We observed an increase in the number of CD68+, CD163+, and CD206– macrophages in the follicular fluid of women with tubal infertility, suggesting that these cells may play a role in embryo development. These findings are preliminary and require further research to confirm their significance. M-CSF, macrophage colony stimulating factor; GM-CSF, granulocyte macrophage colony-stimulating factor; LPS, lipopolysaccharide; INF γ, Interferon gamma; IL-10, Interleukin-10; IL-4, Interleukin-4; IL-13, Interleukin-13; IL-1, Interleukin-1; IL-12, Interleukin-12; IL-23, Interleukin-23; IL-17, Interleukin-17; IL-1RA, Interleukin-1RA; CCL17-CC, chemokin ligand 17; CCL18-CC, chemokin ligand 18; CXCL10-CXC, chemokin ligand 10; TNF-α, tumor necrosis factor α. Created with BioRender.com.
The subpopulations of the macrophages and monocytes in follicular fluid and blood in patients with infertility depending on the infertility factor.
Monocyte and Macrophage in Follicular Liquid: Predictive Markers of Embryo Quality in Women with Obesity and Infertility
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January 2025

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32 Reads

Background Over the past five years, the pregnancy rate in assisted reproductive technology (ART) programs in Russia has remained relatively stable. The aim of this study was to assess the distribution of monocyte and macrophage subsets in the blood and follicular fluid of infertile women undergoing assisted reproductive technology. Methods The study involved 45 women with a mean age of 35 ± 4.66 years. Monocytes and macrophages were identified using flow cytometry. Results We observed a decrease in the CD68+CD163+CD206+ and the CD68+CD163–CD206+ cells in patients with a body mass index (BMI) >25 by 0.19 times and 6.56 times, respectively, compared to the group with a BMI <25 (p = 0.031). Patients with fair oocyte quality had 3.6 times more oocytes than those with poor quality (p = 0.010). The relative content of CD14+163–206+ monocytes was found to be 24.15 times higher in the follicular fluid of women with poor embryo quality compared to the group with good embryos (p = 0.010). We also noted that the number of oocytes increased in women with male factor infertility (p = 0.020) and those with unspecified infertility when compared to tubal infertility. An increase in the relative content of CD14+163+206+ in the blood was higher in women with other causes of female infertility compared to those with male factor infertility (p = 0.010). The relative content of M2-monocytes (CD14+163+206–) in the blood was 4.38 times higher in women with male factor infertility than in women with unexplained infertility (p = 0.010). Conclusions A critical component of the inflammatory reaction in patients undergoing in vitro fertilization (IVF) involves more than just the activation of pro-inflammatory cells in response to ovarian stimulation. Our research shows that changes in the distribution of monocytes and macrophages can influence embryo implantation success and pregnancy outcomes in women. These processes are influenced by various infertility-related factors, including those mentioned above. However, these findings are preliminary and require further investigation.


Atomized Neutrophil Membrane-coated MOF Nanoparticles for Direct Delivery of Dexamethasone for Severe Pneumonia

January 2025

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1 Read

Background Dexamethasone has proven life-saving in severe acute respiratory syndrome (SARS) and COVID-19 cases. However, its systemic administration is accompanied by serious side effects. Inhalation delivery of dexamethasone (Dex) faces challenges such as low lung deposition, brief residence in the respiratory tract, and the pulmonary mucus barrier, limiting its clinical use. Neutrophil cell membrane-derived nanovesicles, with their ability to specifically target hyper-activated immune cells and excellent mucus permeability, emerge as a promising carrier for pulmonary inhalation therapy. Methods We designed a novel UiO66 metal-organic framework nanoparticle loaded with Dex and coated with neutrophil cell membranes (UiO66-Dex@NMP) for targeted therapy of severe pneumonia. This was achieved by loading Dex into UiO66 pores and subsequently coating with neutrophil membranes for functionalization. Results Drug release experiments revealed UiO66-Dex@NMP to exhibit favorable sustained-release properties. Additionally, UiO66-Dex@NMP demonstrated excellent targeting capabilities both in vitro and in vivo. In a mouse model of lipopolysaccharide (LPS)-induced pneumonia, UiO66-Dex@NMP significantly reduced lung inflammation compared to both the control model and Dex administered via inhalation. Histopathological analysis further confirmed UiO66-Dex@NMP’s ability to alleviate lung tissue damage. Conclusions UiO66-Dex@NMP represents a novel and safe inhaled delivery carrier for Dex, offering valuable insights into the clinical management of respiratory diseases, including severe pneumonia.


ERK1/2 phosphorylation is elevated in diabetic conditions. (A) Western blot analysis of ERK1/2 phosphorylation as well as their total levels in H9C2 cells subjected to high glucose (HG), palmitic acid (PA), or both HG and PA (HG+PA) for 24 and 48 hours. Control (con) were H9C2 cells cultured in DMEM that had 5.5 mM glucose. p38 was used as a loading control. (B) Quantification of ERK1/2 phosphorylation based on Fig. 1A. * p < 0.05 vs. con, and ** p < 0.01 vs. con. (C) Analysis of phosphorylation and total protein levels of ERK1/2 in db/db mouse hearts by a Western blot approach. p38 was used as a loading control. (D) Quantification of ERK1/2 phosphorylation levels based on Fig. 1C. ** p < 0.01 vs. con. (E) Fasting serum glucose level in mice 7 days after receiving 5 consecutive days of either a citrate solution (veh) or STZ. *** p < 0.001 vs. veh. (F) Western blot analysis of phosphorylation and total protein levels of ERK1/2 in the hearts of control (veh) and STZ-treated mice. p38 was used as a loading control. (G) Western blot quantification based on Fig. 1F. *** p <0.001. All the samples in Fig. 1 were biological replicates. ERK1/2, extracellular signal-regulated protein kinases 1 and 2; STZ, streptozotocin.
U0126 inhibition attenuates the diabetic cardiomyopathy. (A) Effect of U0126 on ERK1/2 phosphorylation in H9C2 cells at baseline and 20% FBS stimulation conditions. (B) Quantification of ERK1/2 phosphorylation based on Fig. 2A. * p < 0.05 vs. DMSO; ** p < 0.01 vs. DMSO/FBS; *** p < 0.001 vs. DMSO/FBS. (C) Determining ERK1/2 phosphorylation levels in the hearts of mice treated with either 6% DMSO (veh) or U0126 at different doses for one week. (D) Quantification of ERK1/2 phosphorylation in Fig. 2C. * p < 0.05 vs. veh. (E,F) Fasting glucose measurement and heart/body weight ratios in mice 6 weeks post U0126 treatment. * and # p < 0.05. (G–J) RT-PCR analysis of genes in cardiac hypertrophy (G–I) and fibrosis (J). * and # p < 0.05. (K) Representative Masson’s trichrome images for fibrosis analysis in the mouse heart. Scale bar, 100 µm. (L–O) RT-PCR analysis of genes involved in fatty acid uptake (FACS), fatty acid oxidation (CPT1A), fatty acid metabolism (PPARα), and fatty acid synthesis and storage (PPARγ). * p <0.05; ** and ## p < 0.01. All the samples in Fig. 2 were biological replicates. RT-PCR, real-time polymerase chain reaction.
U0126 inhibition in db/db mice alleviates the diabetic cardiomyopathy. (A) Fasting glucose levels in indicated groups of mice. * and # p < 0.05. (B) Comparison of heart/body weight ratios in all three groups of mice. ** p < 0.05 vs. con. n.s., not significant. (C–F) RT-PCR analysis of marker genes for cardiac hypertrophy and interstitial fibrosis. * and # p < 0.05. (G) Fibrosis comparison among three groups of mouse hearts by Masson’s trichrome staining. Scale bar, 100 µm. (H–K) RT-PCR analysis of fatty acid metabolism genes. * and # p < 0.05; ** and ## p < 0.01. All the samples in Fig. 3 were biological replicates.
Activation of ERK1/2 exacerbates diabetic cardiomyopathy. (A) Comparison of ERK1/2 phosphorylation in the hearts of wild type (WT) and DUSP6/8 double knockout mice (DKO). (B) ERK1/2 phosphorylation quantification based on Fig. 4A. * p < 0.05 vs. WT. (C) Fasting glucose levels in indicated groups of mice 1 week after STZ administration. (D) Comparison of heart/body weight ratios in all four groups of mice. (E) Representative Masson’s trichrome images for fibrosis analysis in the mouse hearts. Scale bar, 100 µm. (F,G) Gene expression analysis for cardiac hypertrophy, fibrosis, and fatty acid metabolism. * p < 0.05. All the samples in Fig. 4 were biological replicates.
ERK1/2 Inhibition Alleviates Diabetic Cardiomyopathy by Suppressing Fatty Acid Metabolism

January 2025

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5 Reads

Background Diabetes mellitus is associated with morphological and functional impairment of the heart primarily due to lipid toxicity caused by increased fatty acid metabolism. Extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) have been implicated in the metabolism of fatty acids in the liver and skeletal muscles. However, their role in the heart in diabetes remains unclear. In this study, we tested our hypothesis that pharmacological inhibition of ERK1/2 alleviates cardiac remodeling in diabetic mice through a reduction in fatty acid metabolism. Methods ERK1/2 phosphorylation in diabetes was determined both in vitro and in vivo. H9C2 cells were subjected to high glucose, high palmitic acid, or both high glucose and palmitic acid. db/db and streptozotocin (STZ)-induced diabetic mice were analyzed for ERK1/2 phosphorylation levels as well as the effects of U0126 treatment on cardiac remodeling. Administration of STZ and U0126 in mice was performed via intraperitoneal injection. Blood glucose levels in mice were measured using a glucometer. Mouse heart total RNAs were purified for reverse transcription. Real-time polymerase chain reaction (PCR) analysis of the messenger ribonucleic acid (mRNA) expression was performed for hypertrophy (ANF, BNP, and βMHC), fibrosis (Col3α1), and fatty acid metabolism genes (PPARα, CPT1A, and FACS). Interstitial fibrosis of the myocardium was analyzed using Masson’s trichrome staining of the paraffin-embedded tissues. Results ERK1/2 phosphorylation was significantly increased in diabetic conditions. Inhibition of ERK1/2 by U0126 in both streptozotocin-induced diabetic mice and db/db mice resulted in a significant reduction in the expression of genes associated with hypertrophy and fibrosis. In contrast, elevated phosphorylation of ERK1/2 in Dusp6/8 knockout (DKO) mice resulted in fibrosis. Mechanistically, ERK1/2 activation enhanced the expression of fatty acid metabolism genes PPARα, CPT1A, and FACS in the heart, which was reversed by U0126 treatment. Conclusion ERK1/2 are potential therapeutic targets for diabetic cardiomyopathy by modulating fatty acid metabolism in the heart.


SUMO-Specific Peptidase 5 Promotes Oesophageal Squamous Cell Carcinoma Growth through the NF-κB-SLC1A3 Axis

January 2025

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5 Reads

Background This study investigates the role of small ubiquitin-like modifier (SUMO)-specific peptidase 5 (SENP5), a key regulator of SUMOylation, in esophageal squamous cell carcinoma (ESCC), a lethal disease, and its underlying molecular mechanisms. Methods Differentially expressed genes between ESCC mouse oesophageal cancer tissues and normal tissues were analysed via RNA-seq; among them, SENP5 expression was upregulated, and this gene was selected for further analysis. Immunohistochemistry and western blotting were then used to validate the increased protein level of SENP5 in both mouse and human ESCC samples. The Kaplan‒Meier method and multivariate analysis were used to analyse the relationship between SENP5 expression and ESCC prognosis. Stable SENP5-knockdown (KD) cell lines and conditional knockout (cKO) mice were established to verify the biological function of SENP5. Further RNA-seq comparisons between short hairpin SENP5 (shSENP5)- and short hairpin negative control (shNC)-transfected ESCC cell lines were conducted, and the nuclear factor kappa B (NF-κB)—SLC1A3 axis was identified through bioinformatics analysis. The correlation of SENP5 with signalling pathway components was validated via real-time quantitative PCR (qPCR), western blotting (WB), and immunoprecipitation. Results Our study revealed that SENP5 was upregulated in human and mouse ESCC samples, and clinical data analysis revealed a correlation between high SENP5 expression and poor patient prognosis. SENP5 knockdown inhibited tumorigenesis and growth in vivo and suppressed the proliferation, migration, and invasion of ESCC cell lines in vitro. Our study also revealed that SENP5 knockdown enhanced the SUMO1-mediated SUMOylation of NF-kappa-B inhibitor alpha (IκBα), thereby inhibiting the activation of the NF-κB–SLC1A3 axis, which subsequently suppresses ESCC cell energy metabolism and impedes ESCC progression. Conclusions Suppression of SENP5 slows the development of ESCC by inhibiting the NF-κB‒SLC1A3 axis through SUMO1-mediated SUMOylation of IκBα. Our research suggests that SENP5 could serve as a prognostic indicator and a target for therapeutic intervention for ESCC patients.


HP1 promotes centromeric localization of ATRX. (A) Asynchronous HeLa cells were immunostained for interphase cells. Mitotic cells were collected by treated with nocodazole for 3 hours, then underwent cytospin for immunostaining. (B,C) Cells were treated with nocodazole for a duration of 3 hours, after which the shake-off mitotic cells were subjected to immunostaining. The relative enrichment of ATRX was measured in across 10 cells (B) (analyzed using an unpaired t-test). Example images are shown in (C). (D) Immunoblot analysis was performed on asynchronous HeLa and the relevant HP1-TKO clones. (E,F) The stable cell lines specified underwent treatment with nocodazole for a duration of 3 hours, after which the shake-off mitotic cells were subjected to immunostaining. The relative enrichment of ATRX was measured in across 10 cells (E) (analyzed using an unpaired t-test). Representative images are supplied in (F). (G,H) HeLa cells were transiently transfected with either centromere pro- tein B (CB)-GFP or CB-HP1α-GFP, and the shake-off mitotic cells arrested with nocodazole were subjected to immunostaining. The ratio of centromeric ATRX/GFP immunofluorescence intensity was determined in across 10 different cells (G) (analyzed using an unpaired t-test), and example images are given (H). Data details: The means and standard deviations (SDs) are presented (B,E,G). Scale bars: 10 µm. For further details, refer to Supplementary Fig. 2. ATRX, α thalassemia/mental retardation syndrome X-linked; HP1, heterochromatin protein 1; TKO, triple-knockout; ACA, anti-centromere antibody; GFP, green fluorescent protein.
The N-terminal end of ATRX interacts with HP1α, HP1β, and HP1γ. (A) Nocodazole-arrested mitotic HeLa cell lysates were pulled down using GST or GST-HP1α, and then Coomassie Brilliant Blue staining and anti-ATRX immunoblotting were performed. (B) Lysates of 293T cells arrested in mitosis by nocodazole and transiently expressing HP1α-Flag-6xHis and HA-ATRX were immunoprecipitated using anti-HA antibodies, then analyzed by immunoblotting. (C) Lysates from nocodazole-arrested mitotic HeLa cells transiently expressing HA-ATRX and HP1-Flag-6xHis (including HP1α, HP1β, and HP1γ) were immunoprecipitated using anti-Flag antibodies, which was subsequently followed by immunoblotting with either anti-FLAG or anti-HA antibodies. (D) Mitotic HeLa cell lysates that were arrested with nocodazole underwent immunoprecipitation using anti-ATRX antibodies, and the corresponding antibodies were then used for immunoblotting. (E) Lysates from mitosis HeLa cells, which were treated with nocodazole and transiently expressing SFB-ATRX, were subjected to pulldown assays. This was followed by immunoblotting and staining with Coomassie Brilliant Blue. (F) Lysates from HeLa cells arrested in mitosis with nocodazole were processed through pulldown assays. Following this, immunoblotting and Coomassie Brilliant Blue staining were conducted. (G) Lysates from 293T cells arrested in mitosis with nocodazole and transiently expressing HP1α-Flag and HA-ATRX (both wild type and ΔP1 mutant) were subjected to immunoprecipitation. This was followed by immunoblotting. (H) Lysates from HeLa cells were transiently expressing HP1α-Flag and HA-ATRX (including both wild type and ΔPxVxL mutant) underwent immunoprecipitation. This was followed by immunoblotting. (I) Lysates derived from 293T cells arrested in mitosis with nocodazole were transiently expressing HA-ATRX (either wild type or mutants) were subjected to pulldown assays. Following this, Coomassie Brilliant Blue staining and immunoblotting were conducted. Data information: Please refer to Supplementary Fig. 2 for additional details. GST, Glutathione S-transferase; CBB, Coomassie Brilliant Blue; IP, Immunoprecipitation; HA, hemagglutinin; WT, wild type.
Disruption of the HP1- ATRX interaction delocalizes ATRX from mitotic centromeres, resulting in increased centromeric cohesion defects. (A) Immunoblotting was performed on lysates derived from asynchronous HeLa cells along with the specified stable cell lines. (B,C) The specified stable cell lines as described in A underwent treatment with MG132, with or without the inclusion of endogenous ATRX RNAi cells. Cells were fixed at the given time intervals for DNA staining, and about 200 cells were counted (n = 2). (D) HeLa along with the referenced stable cell lines as described in A were treated with MG132 for 8 hours. The fraction of cells exhibiting cohesion loss was assessed in approximately 200 cells (n = 2) via mitotic chromosome spreads. Sample pictures are shown in Supplementary Fig. 3C. (E,F) The specified cell lines underwent a 3-hour treatment with nocodazole. The immunostaining of mitotic chromosome spreads was performed, and the ratio of immunofluorescence intensity for centromeric HA/CENP-C was measured in across 10 cells (E) (analyzed using an unpaired t-test). Example images are depicted in (F). Data information: Mean values and standard deviations (SDs) are displayed (B,C,E). Scale bar: 10 µm. Refer to Supplementary Fig. 3 for more information. WT, wild type; siControl, small interfering RNA Control.
Depletion of Wapl abolishes the need for the HP1-ATRX interaction in safeguarding centromeric cohesion. (A) Cell lines were transfected with the specified siRNAs were subjected to immunoblotting. (B) The selected cell lines expressing the indicated siRNAs were treated with MG132, fixed at predetermined time points, and analyzed in approximately 200 cells (n = 2). Means and standard deviations (SDs) were recorded. (C,D) HeLa cells and the specified cell lines received MG132 treatment for 8 hours. By utilizing mitotic chromosome spreads, the fraction of cells with cohesion loss was assessed in around 100 cells (C). Example images of the mitotic chromosome spreads are provided (D). Scale bar, 10 µm. (E) Model for HP1 in localizing ATRX at mitotic centromeres, which inhibits Wapl’s release of cohesin from mitotic centromeres.
HP1 Promotes the Centromeric Localization of ATRX and Protects Cohesion by Interfering Wapl Activity in Mitosis

January 2025

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3 Reads

Background α thalassemia/mental retardation syndrome X-linked (ATRX) serves as a part of the sucrose nonfermenting 2 (SNF2) chromatin-remodeling complex. In interphase, ATRX localizes to pericentromeric heterochromatin, contributing to DNA double-strand break repair, DNA replication, and telomere maintenance. During mitosis, most ATRX proteins are removed from chromosomal arms, leaving a pool near the centromere region in mammalian cells, which is critical for accurate chromosome congression and sister chromatid cohesion protection. However, the function and localization mechanisms of ATRX at mitotic centromeres remain largely unresolved. Methods The clustered regularly interspaced short palindromic repeats with CRISPR-associated protein 9 (CRISPR-Cas9) system and overexpression approaches were employed alongside immunofluorescence to investigate the mechanism of ATRX localization at the centromere. To study the binding mechanism between ATRX and heterochromatin protein 1 (HP1), both full-length and truncated mutants of hemagglutinin (HA)-ATRX were generated for co-immunoprecipitation and glutathione S-transferase (GST)-pull assays. Wild-type ATRX and HP1 binding-deficient mutants were created to investigate the role of ATRX binding to HP1 during mitosis, with the Z-Leu-Leu-Leu-al (MG132) maintenance assay, cohesion function assay, and kinetochore distance measurement. Results and Conclusions Our research demonstrated that HP1α, HP1β, and HP1γ facilitate the positioning of ATRX within the mitotic centromere area through their interaction with the first two [P/L]-X-V-X-[M/L/V] (PxVxL)motifs at the N-terminus of ATRX. ATRX deficiency causes aberrant mitosis and decreased centromeric cohesion. Furthermore, reducing Wapl activity can bypass the need for ATRX to protect centromeric cohesion. These results provide insights into the mechanism of ATRX's centromeric localization and its critical function in preserving centromeric cohesion by reducing Wapl activity in human cells.


Androgenic Anabolic Steroids Cause Thiol Imbalance in the Vascular Endothelial Cells

January 2025

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5 Reads

Background Androgenic anabolic steroids (AASs) are synthetic drugs structurally related to testosterone, with the ability to bind to androgen receptors. Their uncontrolled use by professional and recreational sportspeople is a widespread problem. AAS abuse is correlated with severe damage to the cardiovascular system, including changes in homeostasis and coagulation disorders. AASs alter vascular function by blocking nitric oxide (NO)-mediated dilation, impairing endothelial growth and by potentiating vasoconstrictor signals. Methods This paper demonstrated that long-term use of AASs (nandrolone and boldenone), negatively affects the basic cell functions of vascular endothelial cells. The susceptibility of endothelial cells to AASs depends on the expression of androgen receptors, although cells without androgen receptors can also be affected by high doses of AASs to a limited extent. Seven-day incubation with AASs diminishes endothelial cell proliferation and migration (determined by transwell and scratch migration assay) and monolayer formation (using transendothelial electrical resistance assay). Results Disturbances in cell function were accompanied by downregulation of peroxiredoxins (PRDX1 and PRDX2), involved in maintaining the thiol-disulphide balance. In addition, AASs increased oxidation of the non-enzymatic thiol buffer, glutathione (GSH), reduced secretion of thiol oxidoreductase protein disulphide isomerase (PDI) from endothelial cells and affected the thiol pattern of PDI. Conclusions These changes may be related to a thiol-disulfide imbalance and vascular endothelium dysfunction, that are often correlated with abnormal platelet aggregation, inflammation, increased vascular permeability, and vascular smooth muscle cell proliferation—all of which are observed in athletes who abuse AASs.


tiRNA-Gln-CTG is Involved in the Regulation of Trophoblast Cell Function in Pre-eclampsia and Serves as a Potent Biomarker

January 2025

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2 Reads

Background Pre-eclampsia (PE) is a gestational disorder that significantly endangers maternal and fetal health. Transfer ribonucleic acid (tRNA)-derived small RNAs (tsRNAs) are important in the progression and diagnosis of various diseases. However, their role in the development of PE is unclear. Consequently, we detected the expression profiles of tsRNAs in the plasma of patients with PE as well as those in the plasma of the healthy control group, and a multiplicity of experiments were conducted with the aim of clarifying their roles in the occurrence and development of PE and the feasibility of serving as predictive biomarkers for this disorder. Methods High-throughput sequencing of tsRNA in plasma from PE cases was performed to evaluate its potential as a diagnostic or therapeutic biomarker. The function of tsRNA in trophoblasts was explored using the HTR-8/SVneo cell line. Plasma from pregnant women with suspected PE was analyzed to assess the potential of tsRNA to act as a predictive marker of PE. Results High-throughput sequencing of tsRNA was performed on plasma from pregnant women with PE and from healthy pregnant controls. Analysis revealed a significant reduction in the level of tRNA-derived stress-inducing RNA (tiRNA)-Gln-CTG in the plasma (p < 0.001) and placenta (p < 0.001) of pregnant women with PE, suggesting its potential involvement in the development of this condition. tiRNA-Gln-CTG was identified in the cytoplasm and nucleus of HTR-8/SVneo cells. In vitro experiments revealed that tiRNA-Gln-CTG influences the proliferation, cycling, migration, and invasion of HTR-8/SVneo cells, possibly by targeting the 3′UTR region of thrombospondin-2 messenger ribonucleic acid (mRNA) for degradation. Extracellular vesicle (EV) carriers may mediate the level of tiRNA-Gln-CTG in the circulation. Y-box binding protein-1 (YBX1) may be involved in loading tiRNA-Gln-CTG into EVs. The sensitivity of low tiRNA-Gln-CTG levels for predicting the onset of PE in suspected cases was 91.7% within 1 week of delivery, 85.7% within 4 weeks of delivery, and 89.3% before delivery, with corresponding specificities of 84.5%, 79.2%, and 73.4%, respectively. Conclusions tiRNA-Gln-CTG significantly influences trophoblast function and is associated with the development of PE. It can serve as an effective biomarker for predicting PE progression within one week of delivery in women with suspected PE.


Vitamin Bs as Potent Anticancer Agents through MMP-2/9 Regulation

In recent years, the role of coenzymes, particularly those from the vitamin B group in modulating the activity of metalloenzymes has garnered significant attention in cancer treatment strategies. Metalloenzymes play pivotal roles in various cellular processes, including DNA repair, cell signaling, and metabolism, making them promising targets for cancer therapy. This review explores the complex interplay between coenzymes, specifically vitamin Bs, and metalloenzymes in cancer pathogenesis and treatment. Vitamins are an indispensable part of daily life, essential for optimal health and well-being. Beyond their recognized roles as essential nutrients, vitamins have increasingly garnered attention for their multifaceted functions within the machinery of cellular processes. In particular, vitamin Bs have emerged as a pivotal regulator within this intricate network, exerting profound effects on the functionality of metalloenzymes. Their ability to modulate metalloenzymes involved in crucial cellular pathways implicated in cancer progression presents a compelling avenue for therapeutic intervention. Key findings indicate that vitamin Bs can influence the activity and expression of metalloenzymes, thereby affecting processes such as DNA repair and cell signaling, which are critical in cancer development and progression. Understanding the mechanisms by which these coenzymes regulate metalloenzymes holds great promise for developing novel anticancer strategies. This review summarizes current knowledge on the interactions between vitamin Bs and metalloenzymes, highlighting their potential as anticancer agents and paving the way for innovative, cell-targeted cancer treatments.


SERT-Deficient Mice Fed Western Diet Reveal Altered Metabolic and Pro-Inflammatory Responses of the Liver: A Link to Abnormal Behaviors

January 2025

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15 Reads

Background The inheritance of the short SLC6A4 allele, encoding the serotonin transporter (SERT) in humans, increases susceptibility to neuropsychiatric and metabolic disorders, with aging and female sex further exacerbating these conditions. Both central and peripheral mechanisms of the compromised serotonin (5-HT) system play crucial roles in this context. Previous studies on SERT-deficient (Sert-/-) mice, which model human SERT deficiency, have demonstrated emotional and metabolic disturbances, exacerbated by exposure to a high-fat Western diet (WD). Growing evidence suggests the significance of hepatic regulatory mechanisms in the neurobiology of central nervous system disorders, supporting the ‘liver-brain’ concept. However, the relationship between aberrant behavior and hepatic alterations under conditions of SERT deficiency remains poorly investigated. Methods One-year-old female Sert-/- mice and their wild-type (WT) littermates were subjected to a control diet (CD) or the WD for a duration of three weeks. The WD had a higher caloric content and was characterized by an elevated saturated fat content (21%) compared to the CD (4.5%) and contained 0.2% cholesterol. Mice were evaluated for anxiety-like behavior, exploration and locomotor activity in the open field test, as well as glucose tolerance and histological indicators of hepatic steatosis. Hepatic pro-inflammatory and metabolism-related gene expression and markers of nitrosative stress, were analyzed utilizing real-time polymerase chain reaction (RT-PCR) and correlated with behavioral and histological outcomes. Results In comparison to unchallenged mice, Sert-/-/WD mutants, but not the WT/WD group, had increased locomotion and anxiety-like behavior, increased hepatic steatosis, and elevated expression of insulin receptor B and pro-inflammatory cytokines interleukin-1β (Il-1β) and Tnf, as well as decreased expression of leptin receptor B. The two genotypes displayed distinct gene expression patterns of nitric oxide (NO)-related molecules inducible NO synthase (iNos) and arginase (Arg2), insulin receptor-related signaling factors: cluster of differentiation 36 (Cd36), ecto-nucleotide pyrophosphatase/phosphodiesterase (Enpp), protein tyrosine phosphatase N1 (Ptpn1), cytochrome P450 omega-hydroxylase 4A14 (Cyp4a14), acyl-CoA synthetase 1 (Acsl1) and phosphatase and tensin homolog (Pten). Furthermore, there were profound differences in correlations between molecular, histological, and behavioral measurements across the two genotypes. Conclusions Our findings suggest that the genetic deficiency of SERT results in abnormal hepatic pro-inflammatory and metabolic adaptations in response to WD. The significant correlations observed between behavioral measures and pro-inflammatory and metabolic alterations in WD-fed mice suggest the importance of liver-brain interactions and their role in the aberrant behaviors exhibited by Sert-/- mutants. This study presents the first evidence that altered liver functions are associated with pathological behaviors arising from genetic SERT deficiency.



Flowchart of the literature search.
Potential mechanisms of action of winery by-products in atherothrombosis. Modulation of platelet function mediated by winery by-product constituents through inhibition of aggregation factors binding to their membrane receptors, inhibition of signal transduction enzymes and free radicals scavenging. AA, arachidonic acid; AC, adenylyl cyclase; ADP, adenosine diphosphate; ATP, adenosine triphosphate; Ca, calcium; cAMP, cyclic AMP; COX, cyclooxygenase; DAG, diacylglycerol; GP, glycoprotein; IP3, inositol trisphosphate; LDL, low density lipoprotein; oxLDL, oxidized LDL; Ox-PLs, oxidized phospholipids; PAF, platelet activating factor; PAFR, PAF receptor; PAR, protease-activated receptor; PI3K, phosphoinositide-3 kinase; PIP2, phosphatidylinositol biphosphate; PK, protein kinase; PL, phospholipase; P2Y, platelet receptor for ADP; TP, thromboxane receptor; TXA2, thromboxane A2; vWF, von Willebrand factor. Created with BioRender.com (License: OM23Z01REU).
Winery By-Products In Vitro and In Vivo Effects on Atherothrombotic Markers: Focus on Platelet-Activating Factor

Platelet aggregation and inflammation play a crucial role in atherothrombosis. Wine contains micro-constituents of proper quality and quantity that exert cardioprotective actions, partly through inhibiting platelet-activating factor (PAF), a potent inflammatory and thrombotic lipid mediator. However, wine cannot be consumed extensively due to the presence of ethanol. Alternatively, winery by-products are abundant in similar-to-wine micro-constituents that could be used in food fortification and dietary supplements. Also, the vinification process produces millions of tons of by-products worldwide, posing an environmental matter of waste management. Therefore, the purpose of this literature review is to update the existing data concerning the in vitro anti-platelet and anti-inflammatory properties of winery by-product extracts and their possible health effects through controlled clinical trials in humans, specifically focused on their effects on PAF’s actions. Data from in vitro studies report that winery by-product compounds are able to inhibit platelet aggregation against several aggregation factors, as well as to downregulate inflammatory markers. Among their actions, extracts or phenolic compounds present in winery by-products inhibit PAF’s actions, a potent inflammatory and thrombotic mediator. Similar conclusions have been drawn from human supplementation studies, which suggest that winery by-product extracts may have beneficial biological effects on the cardiovascular system. Evidence from long-term studies shows that consumption may lower total and low density lipoprotein (LDL) cholesterol, improve insulin sensitivity, decrease lipid and protein oxidative damage, enhance antioxidant capacity, and have mild anti-inflammatory action toward reducing cytokine expression and levels. Data from the limited postprandial studies report that the acute consumption of winery by-product extracts improves glycemic response and reduces platelet reactivity to aggregatory stimuli. Although wine extracts and phenolic compounds have been reported to inhibit PAF’s actions and reduce the activity of its biosynthetic enzymes, no data exist concerning the influence of winery by-product extracts. In the future, additional long-term randomized controlled trials or postprandial studies are needed to draw definitive conclusions and establish a viable cardioprotective strategy that incorporates the sustainable use of winery by-products.


Cellular Cholesterol Loss Impairs Synaptic Vesicle Mobility via the CAMK2/Synapsin-1 Signaling Pathway

January 2025

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9 Reads

Background Neuronal cholesterol deficiency may contribute to the synaptopathy observed in Alzheimer’s disease (AD). However, the underlying mechanisms remain poorly understood. Intact synaptic vesicle (SV) mobility is crucial for normal synaptic function, whereas disrupted SV mobility can trigger the synaptopathy associated with AD. In this study, we investigated whether cellular cholesterol deficiency affects SV mobility, with the aim of identifying the mechanism that links cellular cholesterol loss to synaptopathy in AD. Methods Lentiviruses carrying 3β-hydroxysteroid-Δ24 reductase-complementary DNA (DHCR24-cDNA), DHCR24-short hairpin RNA (DHCR24- shRNA) or empty lentiviral vectors were transfected into SHSY-5Y cells in order to construct DHCR24 knock-down and knock-in models, along with corresponding controls. Filipin III cholesterol staining was employed to visualize membrane and intracellular cholesterol in the different cell models, and fluorescence intensity was assessed using confocal microscopy. Additionally, we performed immunoblotting to quantify the expression of DHCR24, total calmodulin-dependent protein kinase 2 (CAMK-2), p-CAMK2 (T286), caveolin-1, total synapsin-1, phosphorylated synapsin-1 (p-synapsin-1; S605), and synaptophysin in each experimental group. Results In DHCR24-silenced cells, the loss of cellular cholesterol caused by knock-down of DCHR24 resulted in a significant decrease in the levels of phosphorylated CAMK2 (p-CAMK2) and phosphorylated synapsin-1 (p-synapsin-1) compared to control cells. The reduction in p-CAMK2 and p-synapsin-1 could disrupt SV mobility, thereby reducing replenishment of the readily releasable pool (RRP) from the reserve pool (RP). Furthermore, cells with DHCR24 knock-down showed downregulation of caveolin-1, a crucial lipid raft marker, compared to control cells. Conversely, elevated cellular cholesterol levels caused by knock-in of DHCR24 reversed the effects of cholesterol deficiency, suggesting that CAMK2-mediated synapsin-1 phosphorylation may be regulated in a lipid raft-associated manner. Additionally, we found that cellular cholesterol loss could significantly downregulate the expression of synaptophysin protein, which is vital for SV biogenesis and synaptic plasticity. Conclusion These results suggest that depletion of cellular cholesterol following knock-down of DHCR24 can decrease synaptophysin protein expression and impair SV mobility by regulating the CAMK2-meditated synapsin-1 phosphorylation pathway, potentially via a lipid raft-associated mechanism. Our study indicates a critical role for cellular cholesterol deficiency in AD-related synaptopathy, thus highlighting the potential for targeting cellular cholesterol metabolism in therapeutic strategies.


The pathogenesis of type 1 and type II endometrial cancer. PTEN, phosphatase, and tensin homolog deleted on chromosome ten; hMLH1, human mutl homolog 1; hMSH6, heterodimer of MutS homolog 6; HER-2, human epidermal growth factor 2. Created with Adobe illustrator 2023 (Adobe Inc., San Jose, CA, USA).
New molecular subtypes of endometrial cancer as defined by the World Health Organization (WHO). Based on molecular oncology research, the WHO classified Endometrial Cancer (EC) into four subtypes: Polymerase Epsilon (POLE) mutant type, MSI-H/MMR-d type, NSMP Type, and P53abn type/TP53 mutant type. MSI-H, high microsatellite instability; MMR-d, mismatch repair deficiency; NSMP, non-specific molecular profile; P53abn, Abnormal p53. Created with Adobe illustrator 2023 (Adobe Inc., San Jose, CA, USA).
The role of p53 protein. The p53 protein plays a crucial role in a host of physiological processes, including the promotion of angiogenesis, promotion of cell apoptosis, regulation of metabolism, and the regulation of DNA repair. Created with Adobe illustrator 2023 (Adobe Inc., San Jose, CA, USA).
A Primer on the Role of TP53 Mutation and Targeted Therapy in Endometrial Cancer

Endometrial Cancer (EC) is one of the most common gynecological malignancies, ranking first in developed countries and regions. The occurrence and development of EC is closely associated with genetic mutations. TP53 mutation, in particular, can lead to the dysfunction of numerous regulatory factors and alteration of the tumor microenvironment (TME). The changes in the TME subsequently promote the development of tumors and assist in immune escape by tumor cells, making it more challenging to treat EC and resulting in a poor prognosis. Therefore, it is important to understand the effects of TP53 mutation in EC and to conduct further research in relation to the targeting of TP53 mutations. This article reviews current research progress on the role of TP53 mutations in regulating the TME and in the mechanism of EC tumorigenesis, as well as progress on drugs that target TP53 mutations.



Thyroid Hormone Activation Regulates the Crosstalk between Breast Cancer and Mesenchymal Stem Cells

January 2025

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26 Reads

Background Thyroid Hormones (THs) critically impact human cancer. Although endowed with both tumor-promoting and inhibiting effects in different cancer types, excess of THs has been linked to enhanced tumor growth and progression. Breast cancer depends on the interaction between bulk tumor cells and the surrounding microenvironment in which mesenchymal stem cells (MSCs) exert powerful pro-tumorigenic activities. Methods Primary human MSCs from healthy female donors were co-cultured with DIO2 knock out (D2KO) and wild type (WT) MCF7 breast cancer cells to assess cell growth, migration, invasion and the expression of known epithelial-mesenchymal transition (EMT)- and inflammation-related markers. Furthermore, a surgery-free intraductal delivery model, i.e., the Mouse-INtraDuctal (MIND) injection method, was used as a tool for in vivo characterization of breast tumor formation and progression. Results In this study, we uncovered a novel role of THs in regulating the tumor-stroma crosstalk. MCF7 cells enhanced the intracellular activation of THs through the TH-activating enzyme, D2, fostering their EMT properties and the dialogue with MSCs. D2 inactivation reduced the invasiveness of MCF7 cells and their responsiveness to the pro-tumorigenic induction via MSCs, both in vivo and in vitro. Conclusions Thus, we argue that intracellular activation of THs via D2 is a critical requirement for invasive and metastatic conversion of breast cancer cells, advising the blocking of D2 as a potential therapeutic tool for cancer therapy.


The Role of NF-κB/MIR155HG in Regulating the Stemness and Radioresistance in Breast Cancer Stem Cells

January 2025

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3 Reads

Background Breast cancer stem cells (BCSCs) are instrumental in treatment resistance, recurrence, and metastasis. The development of breast cancer and radiation sensitivity is intimately pertinent to long non-coding RNA (lncRNA). This work is formulated to investigate how the lncRNA MIR155HG affects the stemness and radioresistance of BCSCs. Methods Effects of MIR155HG knockdown on BCSCs were gauged in MCF-7 and MDA-MB-231 cell lines. MIR155HG expression was manipulated in cells, followed by an assessment of stemness, DNA damage repair, apoptosis, cell cycle, and the Wnt signaling pathway under radiation conditions. The interaction between nuclear factor kappa B (NF-κB) subunit RelA and MIR155HG was examined using a dual-luciferase reporter assay. To examine the binding interaction between RelA and MIR155HG promoter, chromatin immunoprecipitation was performed. Results Breast cancer-derived stem cells exhibited a high level of MIR155HG. Knockdown of MIR155HG reduced stemness, enhanced radiosensitivity, induced apoptosis, and arrested cells in the G1 phase. Mechanistically, MIR155HG knockdown repressed Wnt/β-catenin signaling and mediated apoptosis-related protein expressions. NF-κB subunit RelA transcriptionally activated MIR155HG, thereby contributing to radioresistance in BCSCs. Conclusion NF-κB regulates MIR155HG transcriptionally to activate the Wnt pathway, thus enhancing stemness and radioresistance in BCSCs. Targeting MIR155HG may enhance the susceptibility of cancer stem cells to radiation-induced cell death, potentially improving therapeutic outcomes. These findings underscore MIR155HG as a promising therapeutic target for breast cancer.


Trimetazidine: Activating AMPK Signal to Ameliorate Coronary Microcirculation Dysfunction after Myocardial Infarction

Background Myocardial ischemia-reperfusion (I/R) injury and coronary microcirculation dysfunction (CMD) are observed in patients with myocardial infarction after vascular recanalization. The antianginal drug trimetazidine has been demonstrated to exert a protective effect in myocardial ischemia-reperfusion injury. Objectives This study aimed to investigate the role of trimetazidine in endothelial cell dysfunction caused by myocardial I/R injury and thus improve coronary microcirculation. Methods The myocardial I/R mouse model was established, and trimetazidine was administered for 7 days before myocardial I/R model establishment. Echocardiography, 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin-eosin (H&E) staining, and thioflavin S staining were applied to assess myocardial injury and microvascular function. Additionally, the oxygen-glucose deprivation/reperfusion (OGD/R) model was developed in endothelial cells to simulate myocardial I/R injury in vitro. Griess reaction method, immunofluorescence, and western blotting (WB) were employed to detect the expressions of nitric oxide (NO), platelet endothelial cell adhesion molecule-1 (CD31) and vascular endothelial (VE)-cadherin, zonula occludens protein 1 (ZO-1), occludin, vascular endothelial growth factor (VEGF) and adenosine monophosphate (AMP)-activated protein kinase (AMPK) signaling-related proteins in endothelial cells and mouse cardiomyocytes. AMPK pathway inhibitor compound C was used for further mechanism validation. Results Our research demonstrated that trimetazidine can alleviate myocardial pathological injury and cardiac function injury during myocardial I/R. Trimetazidine was observed to improve microvascular reflux phenomenon and microvascular function and barrier injury in myocardial I/R and OGD/R models. Additionally, the expressions of AMPK signal-related proteins were found to be inhibited in myocardial I/R and OGD/R models, which were then activated in mice administered trimetazidine. However, the effects of trimetazidine on endothelial cell function and barrier damage were attenuated after co-treatment with compound C and trimetazidine. Conclusion Trimetazidine ameliorated myocardial I/R-induced CMD by activating AMPK signaling.


The multilayer structure of the epidermis. The image shows multiple layers of the epidermis, the keratinocytes and melanocytes, and part of the dermis.
Wave-length-dependent ultraviolet (UV) radiation penetrates human skin and activates the production of vitamin D3 from the precursor 7-dehydrocholesterol (7-DHC).
The characteristics of young and aged skin as visible to the naked eye (left side: young; right side: aged). Adapted from [44].
The impact of Vitamin D against intrinsic and extrinsic skin aging and age-associated skin diseases. ECM, extracellular matrix; ROS, reactive oxygen species; UVA, ultraviolet A; UVB, ultraviolet B; TIMP, tissue inhibitor metalloproteinase; MMP, matrix metalloproteinase; NF-κB, nuclear factor kappa B.
Common dermatological diseases among the elderly.
Impact of Vitamin D on Skin Aging, and Age-Related Dermatological Conditions

January 2025

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71 Reads

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2 Citations

Human skin is a physical and biochemical barrier that protects the internal body from the external environment. Throughout a person’s life, the skin undergoes both intrinsic and extrinsic aging, leading to microscopic and macroscopic changes in its morphology. In addition, the repair processes slow with aging, making the older population more susceptible to skin diseases. Intrinsic factors associated with advanced age gradually degrade the dermal collagen matrix, resulting in fine wrinkles and reduced elasticity; this is accelerated in post-menopausal women due to estrogen deficiency. In contrast, extrinsic factors associated with advanced age, primarily caused by exposure to ultraviolet (UV) radiation, lead to coarse wrinkles, solar elastosis, hyperkeratosis, irregular pigmentation, and skin cancers. UVB radiation, while contributing to skin photo-aging, also induces the cutaneous synthesis of vitamin D. Vitamin D, in turn, protects the skin from oxidative stress, inflammation, and DNA damage, thereby delaying both chronological and photo-aging. Moreover, research has demonstrated an association between lower vitamin D levels and a higher prevalence of certain cutaneous diseases. This review explores and summarizes the critical role of vitamin D in skin aging and age-related skin diseases. The data presented highlight the importance of maintaining vitamin D adequacy throughout life.


Hydroxyapatite Chitosan Gradient Pore Scaffold Activates Oxidative Phosphorylation Pathway to Induce Bone Formation

January 2025

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2 Reads

Background In this study, we prepared a porous gradient scaffold with hydroxyapatite microtubules (HAMT) and chitosan (CHS) and investigated osteogenesis induced by these scaffolds. Methods The arrangement of wax balls in the mold can control the size and distribution of the pores of the scaffold, and form an interconnected gradient pore structure. The scaffolds were systematically evaluated in vitro and in vivo for biocompatibility, biological activity, and regulatory mechanisms. Results The porosity of the four scaffolds was more than 80%. The 50% and 70% HAMT-CHS scaffolds formed an excellent gradient pore structure, with interconnected pores. Furthermore, the 70% HAMT-CHS scaffold showed better anti-compressive deformation ability. In vitro experiments indicated that the scaffolds had good biocompatibility, promoted the expression of osteogenesis-related genes and proteins, and activated the oxidative phosphorylation pathway to promote bone regeneration. Eight weeks after implanting the HAMT-CHS scaffold in rat skull defects, new bone formation was observed in vivo by micro-computed tomographic (CT) staining. The obtained data were statistically analyzed, and the p-value < 0.05 was statistically significant. Conclusion HAMT-CHS scaffolds can accelerate osteogenesis in bone defects, potentially through the activation of the oxidative phosphorylation pathway. These results highlight the potential therapeutic application of HAMT-CHS scaffolds.


Oxidative and Excitatory Neurotoxic Stresses in CRISPR/Cas9-Induced Kynurenine Aminotransferase Knockout Mice: A Novel Model for Despair-Based Depression and Post-Traumatic Stress Disorder

January 2025

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70 Reads

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1 Citation

Backgrounds Memory and emotion are especially vulnerable to psychiatric disorders such as post-traumatic stress disorder (PTSD), which is linked to disruptions in serotonin (5-HT) metabolism. Over 90% of the 5-HT precursor tryptophan (Trp) is metabolized via the Trp-kynurenine (KYN) metabolic pathway, which generates a variety of bioactive molecules. Dysregulation of KYN metabolism, particularly low levels of kynurenic acid (KYNA), appears to be linked to neuropsychiatric disorders. The majority of KYNA is produced by the aadat (kat2) gene-encoded mitochondrial kynurenine aminotransferase (KAT) isotype 2. Little is known about the consequences of deleting the KYN enzyme gene. Methods In CRISPR/Cas9-induced aadat knockout (kat2-/-) mice, we examined the effects on emotion, memory, motor function, Trp and its metabolite levels, enzyme activities in the plasma and urine of 8-week-old males compared to wild-type mice. Results Transgenic mice showed more depressive-like behaviors in the forced swim test, but not in the tail suspension, anxiety, or memory tests. They also had fewer center field and corner entries, shorter walking distances, and fewer jumping counts in the open field test. Plasma metabolite levels are generally consistent with those of urine: antioxidant KYNs, 5-hydroxyindoleacetic acid, and indole-3-acetic acid levels were lower; enzyme activities in KATs, kynureninase, and monoamine oxidase/aldehyde dehydrogenase were lower, but kynurenine 3-monooxygenase was higher; and oxidative stress and excitotoxicity indices were higher. Transgenic mice displayed depression-like behavior in a learned helplessness model, emotional indifference, and motor deficits, coupled with a decrease in KYNA, a shift of Trp metabolism toward the KYN-3-hydroxykynurenine pathway, and a partial decrease in the gut microbial Trp-indole pathway metabolite. Conclusions This is the first evidence that deleting the aadat gene induces depression-like behaviors uniquely linked to experiences of despair, which appear to be associated with excitatory neurotoxic and oxidative stresses. This may lead to the development of a double-hit preclinical model in despair-based depression, a better understanding of these complex conditions, and more effective therapeutic strategies by elucidating the relationship between Trp metabolism and PTSD pathogenesis.


MG-560 staining patterns and dynamics of sperm. (A) Representative fluorescent images of sperm staining patterns with MG-560 after incubation. These patterns were classified into five groups: whole stained pattern (HMT), head and midpiece stained pattern (M), head-only stained pattern (H), midpiece-only stained pattern (M), and unstained pattern (negative). The scale bar represents 10 µm. (B) The stacked bar graphs represent sperm staining patterns with MG-560 after each incubation time. The percentage of each stained pattern was calculated using the following equation: number of sperm with each stained pattern/total number of examined sperm × 100. Values are expressed as means (n = 5). No significant differences were observed between the groups. (C) Representative images of sperm from the testis (left) and cauda epididymis (right) stained with MG-560, merged with Hoechst 33342 images. The scale bar represents 10 µm. (D) The graph shows a comparison of sperm staining patterns with MG-560 between testicular and cauda epididymal sperm. Testicular sperm were treated as immature, and cauda epididymal sperm as mature. The percentage was calculated for each group using the following equation: number of MG-560 positive or negative sperm/total number of examined sperm × 100. Data are presented as mean ± SEM (n = 4).
MG-560 pre-staining patterns and their dynamic changes in sperm. (A) Dynamics of incubation time-dependent staining patterns in MG-560 pre-stained sperm. Sperm were stained with MG-560 immediately after collection and incubated under capacitation conditions. Stacked bar graphs display the proportions of staining patterns of pre-stained sperm at each incubation time. Staining patterns were categorized as shown in Fig. 1A. The percentage of sperm-stained pattern was calculated using the following equation: no. of each stained pattern sperm/total no. of examined sperm × 100. Values are given as means (n = 5). Different characters show significant difference (p < 0.05). (B) Acrosome-intact and acrosome-reacted sperm were detected by PNA-Lectin-FITC staining and observed using fluorescence microscopy. FITC-negative stained sperm are defined as ‘intact sperm’. FITC-positive partially stained sperm are referred to as ‘partial-AR’, while sperm with FITC positivity over the whole sperm head are referred to as ‘complete-AR’. The scale bar represents 5 µm. (C) Stacked bar graphs display the proportions of sperm with each status of acrosome reaction (AR). The percentage of each acrosome status was calculated using the following equation: number of sperm in each acrosome status/total number of examined sperm × 100. Values are presented as means (n = 5). Different characters show significant difference (p < 0.05). (D) The images are merged images of PNA-Lectin-FITC and MG-560, highlighting HM- and M-stained sperm with complete acrosome reaction (AR), which were the most common at 120 minutes. The scale bar represents 10 µm. (E) The bar graphs represent the proportions of sperm with each MG-560 pre-staining pattern exhibiting complete acrosome reaction (AR) at each incubation time. The percentage of complete-AR sperm in each stained-pattern was calculated by the equation: no. of complete-AR sperm in each stained-pattern/total no. of examined sperm × 100. Values are given as means ± SEM (n = 5). Statistical significance was assessed using Tukey’s test for multiple comparisons. *p < 0.005, **p = 0.0001. (F) The bar graphs show the percentage of acrosome status for each MG-560 staining pattern. Sperm were stained with MG-560 after 120 minutes of incubation. The number of sperm examined is indicated for each MG-560 staining pattern. The percentage of acrosome status was calculated using the following equation: number of sperm in each stained pattern with complete acrosome reaction (AR)/total number of examined sperm × 100. Values are presented as means (n = 3).
Dynamics of MG-560-stained sperm in the presence of cumulus-oocyte complexes (COCs). (A) In vitro fertilization (IVF) was performed using sperm stained with MG-560. After pre-staining with MG-560 and incubating for 60 minutes to allow capacitation, these sperm were inseminated without and with cumulus-oocyte complexes (‘incubation’, ‘IVF’ group, respectively). Sixty minutes after insemination, sperm were collected and observed using fluorescence microscopy. Stacked bar graphs display the percentage of each stained patterns in incubation and IVF group. Values are given as means (n = 4). Statistically significant differences were identified between the percentages of HM- and M-patterns in the two groups. Different characters show significant difference (p < 0.05). (B) The bar graph shows the percentage of complete-AR sperm in examined sperm in two groups. The percentage of complete-AR was calculated by the equation: no. of complete-AR sperm/total no. of examined sperm × 100. Values are given as means ± SEM (n = 4). No statistical significance (n.s.) was found between the two groups. (C) Stacked bar graphs display the proportions of MG-560-stained sperm patterns exhibiting complete acrosome reaction (AR) in the incubation and IVF groups. The percentage of each stained pattern with complete AR was calculated using the following equation: number of sperm with each stained pattern and complete AR/total number of complete AR sperm × 100. Values are presented as means (n = 4). Statistically significant differences were identified between the percentages of HM- and M-patterns in the two groups. Different characters show significant difference (p < 0.05).
MG-560 staining patterns of sperm attached to the zona pellucida and within the perivitelline space. (A) Sixty minutes after insemination into cumulus-oocyte complexes (COCs), sperm attached to the zona pellucida and within the perivitelline space were observed. MG-560 pre-stained sperm were found binding to the zona pellucida (ZP) (left) or in the perivitelline space (right). Scale bar represents 10 µm. (B) IVF was performed using denuded oocytes. MG-560 pre-stained sperm were again found binding to the ZP (left) or in the perivitelline space (right). Note that H-, M-, and negative-stained patterns are indicated as representatives. Scale bar represents 10 µm. (C) The graph represents the proportions of pre-staining patterns of sperm binding to ZP or found in the perivitelline space. The data in the graph are shown for both conditions: after IVF in the presence or absence of COCs. The number of examined sperm in each group is indicated as (n). The percentage of each stained pattern was calculated using the equation: number of sperm in each stained pattern/total number of examined sperm × 100. Values are presented as means (n = 5). Different characters show significant difference (p < 0.05).
Heterogeneity in Fluorescence-Stained Sperm Membrane Patterns and Their Dynamic Changes Towards Fertilization in Mice

January 2025

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21 Reads

Background Sperm represent a heterogeneous population crucial for male reproductive success. Additionally, sperm undergo dynamic changes during maturation and capacitation. Despite these well-established processes, the complex nature of sperm heterogeneity and membrane dynamics remains elusive. The composition of phospholipids in the sperm membrane changes dynamically during maturation, with their release occurring during capacitation. This study aims to investigate the heterogeneity and dynamic changes in the sperm membrane during maturation and capacitation towards fertilization by visualizing these membrane dynamics. Methods Sperm were collected from the cauda epididymis or testis of Institute of Cancer Research (ICR) male mice and stained with MemBright dye (commercial name: MemGlow™-560, MG-560), a fluorogenic live-cell membrane probe. Staining was performed either before, during, or after incubation for capacitation. In pre-staining experiments, sperm were stained with MG-560 before capacitation and then incubated to induce capacitation. Acrosome-reacted sperm were assessed after staining with peanut agglutinin FITC (PNA-Lectin FITC). Stained sperm were observed using fluorescence or confocal microscopy. Results MG-560-stained sperm from the epididymis before capacitation showed four staining patterns: head-midpiece-tail (HMT), head-midpiece (HM), head (H), midpiece (M) positive, or totally negative, with ratios remaining unchanged during capacitation (30.5%, 29%, 11.3%, 3.7%, and 25.5%, respectively). In contrast, all testicular sperm were negative for staining. Pre-stained sperm exhibited an increased number of HM and M patterns over time, whereas the number of HMT-stained sperm decreased. Consistently, spontaneous acrosome-reacted sperm were detected predominantly in HM- or M-stained sperm. After in vitro fertilization (IVF) using pre-stained sperm, zona pellucida-attached sperm were mostly negative for staining. Finally, all sperm detected in the perivitelline space were only negative. Conclusions Mature sperm membranes stained with MG-560 exhibited heterogeneous and dynamic changes during the capacitation and fertilization process. MG-560 staining identified sperm with the potential to undergo the acrosome reaction, and these MG-560-positive sperm eventually became negative as they penetrated the zona pellucida for fertilization. Thus, the MG-560 staining patterns likely reflect the physiological state and potential of the sperm. These findings provide new insights into sperm heterogeneity and dynamics, and this staining method may also prove useful for assessing sperm quality.


The Formation and Features of Massive Vacuole Induced by Nutrient Deficiency in Human Embryonic Kidney Cells

January 2025

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11 Reads

Background Cellular vacuolization is a commonly observed phenomenon under physiological and pathological conditions. However, the mechanisms underlying vacuole formation remain largely unresolved. Methods LysoTracker Deep Red probes and Enhanced Green Fluorescent Protein-tagged light chain 3B (LC3B) plasmids were employed to differentiate the types of massive vacuoles. By confocal microscopy, lysosome-like massive vacuoles (LysoTracker Deep Red⁺), autophagosome-like massive vacuoles (LC3B-enhanced green fluorescent protein (EGFP⁺)), and autolysosome-like massive vacuoles (LC3B-EGFP⁺ LysoTracker Deep Red⁺) in starved HEK293T cells were observed. Results In this study, we demonstrated that nutrient deficiency can induce the formation of massive vacuoles that appear highly electron-lucent in HEK293T cells. Additionally, these massive vacuoles, resulting from nutrient depletion, can originate from various organelles, including small vacuoles, autophagosomes, lysosomes, and autolysosomes. We found that massive vacuoles could form through two primary mechanisms: the accumulation of small vacuoles into larger vacuoles or the fusion of homogeneous or heterogeneous vacuoles. Further analysis revealed that the membranes of massive vacuoles, regardless of origin, were composed of a bilayer membrane structure. As the volume of the massive vacuoles increased, the cytoplasm and nucleus were displaced toward the periphery of the cells, leading to the formation of signet ring-like cells. Interestingly, we provided evidence that complete massive vacuoles or autophagosome-like massive vacuoles can be released and exist independently outside HEK293T cells. Conclusions Nutrient deprivation induces the formation of heterogeneous, massive vacuoles in human embryonic kidney cells, some of which contribute to the development of signet ring cells, while others lead to extracellular vacuole formation.


Mechanistic role of MSI2 in HCC. MSI2 promotes the proliferation of myofibroblasts and directly regulates the EGF, TGF-β, Notch signaling pathway and Wnt1-promoted EMT process, which in turn promotes the proliferation of tumor cells; MSI2 regulates hepatocellular carcinoma SCs. RRM, RNA recognition motif; PPD, protein-protein binding domain; HCC, hepatocellular carcinoma; EGF, epidermal growth factor; TGF-β, transforming growth factor beta; EMT, epithelial-mesenchymal transition; SCs, stem cells; CD44v6, cluster of differentiation 44 variant 6.
Mechanistic role of MSI2 in PC and CRC. USP10 promotes MSI2 ubiquitination and the TGF-β/Smad2/3 signaling pathway regulates MSI2 to promote the development of CRC. MSI2 regulates the Numb pathway, p53, ISYNA1, and p21 to promote the proliferation of tumor cells. MSI2 promotes the EMT process and regulates tumor proliferation by regulating the ZEB1-ERK/MAPK signaling pathway. MSI2 directly binds to mRNAs of SAV1 and MOB1 to regulate the Hippo pathway, which promotes the development of PDAC. In the figure, the red line indicates the upward adjustment and the green line indicates the downward adjustment. PC, pancreatic cancer; CRC, colorectal cancer; ISYNA1, Inositol-3-phosphate synthase 1; USP10, ubiquitin-specific protease 10; ZEB1, zinc finger E-box-binding homeobox 1; ERK, extracellular signal-regulated kinase; MAPK, mitogen-activated protein kinase; SAV1, salvador homolog 1; MOB1, Mps one binder 1; PDAC, pancreatic ductal adenocarcinoma; RRM, RNA recognition motif; PPD, protein-protein binding domain.
Mechanism of MSI2 in BC and kidney cancer. In kidney cancer, CD4 and CD8 T cells positively regulate MSI2. MSI2 regulates the JAK2/STAT3 pathway to promote the occurrence and development of BC. MiR-149 and MiR-143 regulate MSI2 to promote the progression of BC. In the figure, “+” indicates an upward adjustment and “-” indicates a downward adjustment. CD4, cluster of differentiation 4; BC, bladder cancer; RCC, renal cell carcinoma; DNACR, differentiation antagonistic non-protein nodal RNA; JAK2/STAT3, Janus tyrosine Kinase 2/Signal Transducer and Activator of Transcription 3.
Update on the Progress of Musashi-2 in Malignant Tumors

Since the discovery of the Musashi (MSI) protein, its ability to affect the mitosis of Drosophila progenitor cells has garnered significant interest among scientists. In the following 20 years, it has lived up to expectations. A substantial body of evidence has demonstrated that it is closely related to the development, metastasis, migration, and drug resistance of malignant tumors. In recent years, research on the MSI protein has advanced, and many novel viewpoints and drug resistance attempts have been derived; for example, tumor protein p53 mutations and MSI-binding proteins lead to resistance to protein arginine N-methyltransferase 5-targeted therapy in lymphoma patients. Moreover, the high expression of MSI2 in pancreatic cancer might suppress its development and progression. As a significant member of the MSI family, MSI2 is closely associated with multiple malignant tumors, including hematological disorders, common abdominal tumors, and other tumor types (e.g., glioblastoma, breast cancer). MSI2 is highly expressed in the majority of tumors and is related to a poor disease prognosis. However, its specific expression levels and regulatory mechanisms may differ based on the tumor type. This review summarizes the research progress related to MSI2 in recent years, including its occurrence, migration mechanism, and drug resistance, as well as the prospect of developing tumor immunosuppressants and biomarkers.


Structure of sulfatides. Sulfatides are composed of galactosylceramide (GalCer) and a polar sulfated carbohydrate sulfate group. Sulfatides contain various structures, including different acyl chain lengths and the ceramide moiety, which can be hydroxylated at the α-2 carbon of the fatty acid. The 2-hydroxyl group is shown in green. Figure created in ChemDraw (RRID: SCR_016768) Professional version 22.0 (PerkinElmer, Waltham, MA, USA).
Synthesis and degradation of sulfatides. Sulfatide synthesis begins in the endoplasmic reticulum by the addition of galactose from UDP-galactose to ceramide, catalyzed by UGT8. After transport of GalCer to the Golgi apparatus, sulfatides are finally synthesized by addition of the sulfate group from PAPS catalyzed by the enzyme GAL3ST1. Sulfatide degradation takes place in the lysosomes, where ARSA hydrolyzes the sulfate group. This reaction requires the action of a non-enzymatic proteinaceous cofactor SapB that removes sulfatides from the membranes and makes accessible to ARSA. GALC catalyzes the hydrolysis of galactose from GalCer to ceramide. UDP-galactose, uridine diphosphate-galactose; UGT8, ceramide galactosyltransferase; GalCer, galactosylceramide; PAPS, 3′-phosphoadenosine-5′-phosphosulfate; GAL3ST1, cerebroside sulfotransferase; ARSA, arylsulfatase A; SapB, saposin B; GALC, galactocerebrosidase. Figure created in ChemDraw (RRID: SCR_016768) Professional version 22.0 (PerkinElmer, Waltham, MA, USA).
Model depicting sulfatide-mediated activation of type II NKT cells reducing liver damage by regulating the proinflammatory cascade of type I NKT cells. Sulfatide activates type II NKT cells, initiating an anti-inflammatory response through the secretion of IL-10, which promotes tolerization in mDCs and inhibits the activation of type I NKT cells. In contrast, type I NKT cells, activated by autolipid or foreign antigens, such as α-GalCer and microbial lipids, or through TLRs/cytokine-mediated pathways, trigger a rapid proinflammatory response involving secretion of OPN, Fas/FasL and IFN-γ. APC, antigen-presenting cells; pDC, plasmacytoid dendritic cells; mDC, myeloid dendritic cells; NKT, natural killer T; IL-10, interleukin-10; IFN-γ, interferon-gamma; TLR, toll-like receptor; OPN, osteopontin; FasL, Fas ligand. Figure created in ChemDraw (RRID: SCR_016768) Professional version 22.0 (PerkinElmer, Waltham, MA, USA).
The Role of Sulfatides in Liver Health and Disease

Sulfatides or 3-O-sulfogalactosylceramide are negatively charged sulfated glycosphingolipids abundant in the brain and kidneys and play crucial roles in nerve impulse conduction and urinary pH regulation. Sulfatides are present in the liver, specifically in the biliary tract. Sulfatides are self-lipid antigens presented by cholangiocytes to activate cluster of differentiation 1d (CD1d)-restricted type II natural killer T (NKT) cells. These cells are involved in alcohol-related liver disease (ArLD) and ischemic liver injury and exert anti-inflammatory effects by regulating the activity of pro-inflammatory type I NKT cells. Loss of sulfatides has been implicated in the chronic inflammatory disorder of the liver known as primary sclerosing cholangitis (PSC); bile ducts deficient in sulfatides increase their permeability, resulting in the spread of bile into the liver parenchyma. Previous studies have shown elevated levels of sulfatides in hepatocellular carcinoma (HCC), where sulfatides could act as adhesive molecules that contribute to cancer metastasis. We have recently demonstrated how loss of function of GAL3ST1, a limiting enzyme involved in sulfatide synthesis, reduces tumorigenic capacity in cholangiocarcinoma (CCA) cells. The biological function of sulfatides in the liver is still unclear; however, this review aims to summarize the existing findings on the topic.


The Impact of Selenium on the Physiological Activity of Yeast Cells Saccharomyces cerevisiae ATCC 7090 and Rhodotorula glutinis CCY 20-2-26

January 2025

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Background This study investigated the selenium-binding capacity of the biomass of two yeast strains, Saccharomyces cerevisiae American Type Culture Collection (ATCC) 7090 and Rhodotorula glutinis CCY 20-2-26. Methods The studies carried out methods of bioaccumulation by yeast biomass. Inorganic selenium was added to the culture media as an aqueous solution of Na2SeO3 at concentrations ranging from 0 to 40 mg Se⁴⁺/L. Results The addition of selenium at concentrations >0.5 mg/L significantly reduced biomass yield compared with the control in the case of S. cerevisiae. A significant reduction in the biomass of R. glutinis was observed only at selenium doses >30 mg/L. The study found that for S. cerevisiae, cultivation should occur for 24 h in a medium with an initial selenium concentration of 20 mg/L to achieve the most efficient selenium accumulation by the yeast biomass. Under these conditions, the yeast could accumulate 4.27 mg Se⁴⁺/g. For the red yeast R. glutinis, optimal selenium binding conditions were achieved by cultivating for 48 h in a medium with an initial selenium ion concentration of 40 mg/L. This yeast strain was more resistant to high selenium doses, accumulating 7.53 mg Se⁴⁺/L at the highest tested dose (40 mg Se⁴⁺/L). Selenium supplementation of the medium from 20 mg Se⁴⁺/L and cultivation for 72 h caused significant changes in the morphology of S. cerevisiae cells (e.g., increased surface area compared with the control). Selenium doses of 20–40 mg/L after 48 h of cultivation significantly reduced the surface area compared with the control results for R. glutinis cells. Conclusions Selenium significantly impacted carotenoid pigment production, with levels decreasing as the selenium concentration in the medium increased. Furthermore, selenium in the tested concentration range increased protein content in the cellular biomass but did not affect intracellular lipid production.


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33.5 days

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