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BACKGROUND: It has been found that abnormal apoptosis of bone tissue cells induced by abnormal iron metabolism plays an important role in the progression of osteoporosis. OBJECTIVE: To investigate the effect of naringin on iron metabolism and cell apoptosis in bone tissue of rats with osteoporosis. METHODS: Fifty 2-month-old female Sprague-Dawley rats were randomly divided into five groups with 10 rats in each group: sham group, osteoporosis group, naringin low-dose group, naringin high-dose group, and naringin high-dose+DKK-1 group. Except for the sham group, rat models of osteoporosis were established by removing bilateral ovarian tissues in the other groups. At 8 weeks after modeling, rats in the naringin low- and high-dose groups were given 100 and 400 mg/kg/d naringenin by gavage, respectively, and rats in the naringenin high dose+DKK-1 group were given 400 mg/kg/d naringin by gavage and subcutaneous injection of 25 mg/kg/d DKK-1, an inhibitor of the Wnt1 signaling pathway, for 7 consecutive days. Relevant indexes were detected after administration. RESULTS AND CONCLUSION: Compared with the osteoporosis group, naringin could enhance the bone mineral density and serum calcium and superoxide dismutase levels in rats (P < 0.05), and reduce the serum levels of osteocalcin, malondialdehyde, and phosphorus (P < 0.05), while DKK-1 could partially inhibit the interventional effect of naringin (P < 0.05). Results from Micro-CT scanning, hematoxylin-eosin and TUNEL staining showed that compared with the osteoporosis group, naringin significantly improved bone microstructure and reduced the rate of cell apoptosis, while DKK-1 partially inhibited the interventional effect of naringin. Immunofluorescence staining results showed that compared with the osteoporosis group, naringin could reduce the oxygen content, anti-tartaric acid phosphatase expression, and elevate the expression of alkaline phosphatase in active tibia tissues (P < 0.05), while DKK-1 could partially inhibit the interventional effect of naringin (P < 0.05). Results from Prussian blue staining and immunohistochemical staining showed that compared with the osteoporosis group, naringin reduced iron deposition in bone and liver tissues as well as the expression of transferrin receptor 1 (P < 0.05), and elevated the protein expression of ferroportin 1 (P < 0.05) in bone tissue, and DKK-1 partially inhibited the intervention of naringin (P < 0.05). PCR and western blot assay of tibia specimens showed that compared with the osteoporosis group, naringin decreased the expression of anti-tartrate acid phosphatase, transferrin receptor 1 and Bax (P < 0.05), and elevated the expression of alkaline phosphatase, ferroportin 1, Bcl-2, Wnt1 and β-catenin (P < 0.05), while DKK- 1 partially inhibited the interfering effect of naringin (P < 0.05). To conclude, naringin inhibits the progression of osteoporosis by reducing iron deposition and apoptosis rate in bone tissue, which may be related to the activation of the Wnt1 signaling pathway. [ABSTRACT FROM AUTHOR]

Objective: To explore the restorative effects and mechanisms of neural stem cell (NSC) transplantation on ischemic brain injury based on the Wnt signaling pathway. Methods: Out of 102 male KM mice, 15 were randomly selected as the control group without any intervention, while the remaining 87 underwent middle cerebral artery occlusion (MCAO) using the Zea-Longa suture method. Seven mice that did not successfully model MCAO were excluded, leaving 80 mice that successfully underwent MCAO, randomized into two groups: the Ischemic Brain Injury group (n = 40) receiving 10 μL of sterile PBS solution injected into the lateral ventricle, and the Ischemic Brain Injury + NSCs Transplantation group (n = 40) receiving 10 μL of NSCs suspension injected into the lateral ventricle. Results: Compared to the ischemic brain injury group, mice in the Ischemic Brain Injury + NSCs Transplantation group exhibited significantly alleviated edema in the middle cerebral artery supply area, with neurons displaying more normal morphological characteristics and fewer signs of degeneration and necrosis. The mice with NSC transplantation had significantly smaller infarct volume than those in the ischemic brain injury group (p < 0.05). The mice with NSC transplantation showed significantly lower Zea-Longa scores and a lower proportion of TUNEL-positive cells compared to those in the ischemic brain injury group (p < 0.05). Conclusion: NSC transplantation can significantly inhibit neuronal apoptosis in the ischemic region of mice with ischemic brain injury, alleviate brain tissue edema, reduce infarct volume, and improve neurological function. The mechanism may be related to Wnt signaling pathway activation. [ABSTRACT FROM AUTHOR]

This study aims to explore the molecular mechanisms of miR-206-3p in regulating Hsp90aa1 and its involvement in the central nervous system (CNS) injury in heat stroke. Weighted gene co-expression network analysis (WGCNA) was performed on the GSE64778 dataset of heat stroke to identify module genes most closely associated with disease characteristics. Through the selection of key genes and predicting upstream miRNAs using RNAInter and miRWalk databases, the regulatory relationship between miR-206-3p and Hsp90aa1 was determined. Through in vitro experiments, various methods, including bioinformatics analysis, dual-luciferase reporter gene assay, RIP experiment, and RNA pull-down experiment, were utilized to validate this regulatory relationship. Furthermore, functional experiments, including CCK-8 assay to test neuron cell viability and flow cytometry to assess neuron apoptosis levels, confirmed the role of miR-206-3p. Transmission electron microscopy, real-time quantitative PCR, DCFH-DA staining, and ATP assay were employed to verify neuronal mitochondrial damage. Heat stroke rat models were constructed, and mNSS scoring and cresyl violet staining were utilized to assess neural functional impairment. Biochemical experiments were conducted to evaluate inflammation, brain water content, and histopathological changes in brain tissue using H&E staining. TUNEL staining was applied to detect neuronal apoptosis in brain tissue. RT-qPCR and Western blot were performed to measure gene and protein expression levels, further validating the regulatory relationship in vivo. Bioinformatics analysis indicated that miR-206-3p regulation of Hsp90aa1 may be involved in CNS injury in heat stroke. In vivo, animal experiments demonstrated that miR-206-3p and Hsp90aa1 co-localized in neurons of the rat hippocampal CA3 region, and with prolonged heat stress, the expression of miR-206-3p gradually increased while the expression of Hsp90aa1 gradually decreased. Further in vitro cellular mechanism validation and functional experiments confirmed that miR-206-3p could inhibit neuronal cell viability and promote apoptosis and mitochondrial damage by targeting Hsp90aa1. In vivo, experiments confirmed that miR-206-3p promotes CNS injury in heat stroke. This study revealed the regulatory relationship between miR-206-3p and Hsp90aa1, suggesting that miR-206-3p could regulate the expression of Hsp90aa1, inhibit neuronal cell viability, and promote apoptosis, thereby contributing to CNS injury in heat stroke. [ABSTRACT FROM AUTHOR]

Introduction: Gliomas, particularly glioblastomas (GBM), are highly aggressive with a poor prognosis and low survival rate. Currently, deoxyelephantopin (DET) has shown promising anti-inflammatory and anti-tumor effects. Using clinical prognostic analysis, molecular docking, and network pharmacology, this study aims to explore the primary targets and signaling pathways to identify novel GBM treatment approaches. Methods: Using PharmMapper, the chemical structure of DET was examined for possible targets after being acquired from PubChem. GBM-related targets were obtained through multi-omics approaches. A protein-protein interaction (PPI) network was constructed using Cytoscape and STRING, and target binding was evaluated through molecular docking. Enrichment analysis was conducted using Metascape. The effects of DET on GBM cell invasion, apoptosis, and proliferation were assessed through in vitro assays, including Transwell, EDU, CCK8, and flow cytometry. Western blot analysis was performed to examine the components of the PI3K/AKT signaling pathway. Results: Among the sixty-four shared targets identified, JUN and CCND1 were the most frequently observed. Enrichment analysis demonstrated that DET influenced the MAPK and PI3K/AKT signaling pathways. In Transwell assays, DET significantly inhibited the invasive behavior of glioma cells. Western blot analysis further confirmed the downregulation of EGFR, JUN, and PI3K/AKT. Conclusion: DET inhibits GBM cell invasion, proliferation, and apoptosis via modulating the PI3K/AKT signaling pathway, highlighting its potential as a novel therapeutic strategy for GBM treatment. [ABSTRACT FROM AUTHOR]

BACKGROUND: Pinoresinol diglucoside promotes bone formation and bone matrix synthesis and accelerates bone tissue repair. However, the mechanism of action and effects of this compound in osteoblasts need to be further explored. OBJECTIVE: To investigate the effect and mechanism of action of pinoresinol diglucoside on dexamethasone-treated osteoblasts based on the Wnt/β-catenin signaling pathway. METHODS: Different concentrations of dexamethasone groups and pinoresinol diglucoside groups were set to treat osteoblasts for 24 hours, and the optimal intervention concentrations were screened. Osteoblasts were treated with dexamethasone, pinoresinol diglucoside and inhibitor XAV-939. Then, control group, dexamethasone group, XVA-939 group, pinoresinol diglucoside group, pinoresinol diglucoside+XVA-939 group were set up. Cell counting kit-8 assay was used to detect cell activity. Alkaline phosphatase activity and caspase3/7 enzyme activity in cells were detected. Annexin V/PI staining and EdU assay were used to detect cell apoptosis and proliferation. Real-time qPCR and western blot were used to detect the mRNA and protein expression levels of Wnt3a, β-catenin, c-myc, osteocalcin, and type I collagen, respectively. RESULTS AND CONCLUSION: After dexamethasone and pinoresinol diglucoside intervened in osteoblasts for 24 hours, 10 μmol/L dexamethasone was found to be the optimal intervention concentration for cell inhibition, and cell proliferation was most pronounced at a concentration of pinoresinol diglucoside of 100 μmol/L. Compared with the dexamethasone group, alkaline phosphatase activity was significantly enhanced (P < 0.05) and caspase3/7 enzyme activity was significantly reduced (P < 0.05) in the pinoresinol diglucoside group. Annexin V/PI staining and cell proliferation assay by EdU method showed that pinoresinol diglucoside inhibited apoptosis and promoted proliferation of osteoblasts after dexamethasone intervention. The mRNA and protein expression levels of Wnt3a, β-catenin, c-myc, osteocalcin, and type I collagen were significantly higher in the pinoresinol diglucoside group and pinoresinol diglucoside+XVA-939 group compared with the dexamethasone and XVA-939 groups (P < 0.05). To conclude, pinoresinol diglucoside can inhibit osteoblast apoptosis after dexamethasone intervention, protect osteoblast activity and promote osteoblast proliferation by activating the Wnt/β-catenin signaling pathway, which may play a role in delaying steroid-induced osteonecrosis of the femoral head . [ABSTRACT FROM AUTHOR]

BACKGROUND: How to improve the hematopoietic microenvironment is a hot topic in the treatment of aplastic anemia. OBJECTIVE: To explore the action mechanism of Angelica sinensis polysaccharide in the treatment of aplastic anemia by combining GEO sequencing analysis, network pharmacology, and experimental validation. METHODS: Aplastic anemia-related differentially expressed genes were obtained from GEO database, and gene ontology and gene set enrichment analysis were performed. The active components and targets of Angelica sinensis polysaccharide were obtained by combining the literature with PubChem, SwissTargetPrediction, and PharmMapper databases. After the intersection targets were taken, STRING and Cytoscape were used to construct proteinprotein interaction network, and gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis was performed. Mouse model of aplastic anemia was established, and the effect and action mechanism of Angelica sinensis polysaccharide on aplastic anemia were verified by blood cell analyzer, flow cytometry, ELISA, immunohistochemistry, immunofluorescence, and western blot assay. RESULTS AND CONCLUSION: (1) A total of 834 differentially expressed genes were screened, which were involved in biological processes such as cell development, hematopoiesis, and myeloid cell differentiation. (2) 347 targets related to Angelica sinensis polysaccharide were retrieved and 77 potential therapeutic genes were screened. Among them, the degree values of angiogenic, apoptotic, and immune-related factors such as VEGFA, EGLN1, Bcl-2, interferon-γ, interleukin-2, interleukin-4, and interleukin-6 were significant. (3) KEGG pathway enrichment analysis revealed that the therapeutic targets were mainly enriched in Th17 cell differentiation, NK-related cytotoxicity, cell adhesion factors such as interferon-γ, interleukin-2, and interleukin-4 related signaling pathways. (4) Animal experiments showed that Angelica sinensis polysaccharide significantly improved bone marrow haematopoiesis, increased peripheral blood cell, and bone marrow single nucleated cell counts, and improved the survival rate of mice. Compared with the model group, mice in the Angelica sinensis polysaccharide group showed a significant decrease in the ratio of Th1/Th2 cells (P < 0.01), a decrease in the expression level of interferon-γ (P < 0.01), an increase in the level of interleukin-4 (P < 0.05), a significant increase in the level of VEGFA (P < 0.01), a significant decrease in EGLN1 (P < 0.01), a significant decrease in apoptosis rate of bone marrow single nucleated cells and reactive oxygen species level (P < 0.01), and a significant increase in cleaved Caspase-3 protein expression (P < 0.01), and a significant decrease in Bcl-2/Bax ratio (P < 0.01). (5) These findings show that Angelica sinensis polysaccharide can improve hematopoiesis of aplastic anemia mice by regulating aberrant T-cell subsets and promoting angiogenesis to improve hematopoietic microenvironment, and inhibiting apoptosis of bone marrow mononuclear cells. [ABSTRACT FROM AUTHOR]

Mitochondrial transcription factor B1 (TFBIM) is mainly involved in mitochondrial DNA transcription and related to the development of hepatocellular carcinoma and breast cancer. However, its role in colon cancer is unclear. In this study, the expression level of TFBIM in colon cancer and its prognosis were analyzed based on TCGA and other databases and IHC assays. Differentially expressed genes (DEGs) were screened and subjected to the analysis of functional enrichment, mutation analysis, immune cell infiltration, and drug sensitivity analysis. CCK-8 and flow cytometry were used to detect the effects of overexpression of TFB1M on the viability, apoptosis and cell cycle distribution of colon cancer cells. Our results showed that the expression level of TFBIM was significantly up-regulated in colon cancer, and its expression level was correlated with the N stage and TNM stage (P<0.05). The prognosis of colorectal cancer patients in the high TFBIM expression group was worse. Functional enrichment results showed that TFBIM was related to leukocyte-mediated immunity, immunoglobulin production and other signaling pathways. Mutation results showed that high-frequency mutated genes, such as ZFHX4, RYR2, PIK3CA and FAT4, had significantly higher mutation frequencies in the TFBIM high-expression group (all P<0.05). In addition, the expression level of TFB1M was significantly higher in the PIK3CA and FAT4 mutation groups (all P<0.05). Immune infiltration results showed that the percentage of CD4 memory activated T cells was increased in the TFB1M high expression group, while the percentage of Treg cells was reduced. The drug sensitivity results showed that patients in the TFBIM high expression group might be more sensitive to Tozasertib, cytarabine, vincristine, etc., while patients in the TFBIM low expression group might be more sensitive to Dasatinib, JQ1, ERK_2440, etc. The results of cellular experiments showed that over-expression of TFB1M enhanced viability, reduced apoptosis and increased the percentage of S-phase and G2/M-phase cells in colon cancer cells. Altogether, the results indicated that TFBIM might play a key role in the growth of colon cancer cells by regulating immune cell infiltration and function. [ABSTRACT FROM AUTHOR]

Background: Among all the gynaecologic malignancies, ovarian cancer (OV) has the highest mortality rate, and herbal plants have been regarded as an alternative strategy for ovarian cancer therapy. Accumulating evidence indicates that the aboveground part of Eclipta prostrata (L.) L., known as Ecliptae Herba (EH), has anti-cancer activity and can contribute to the management of gynaecological disorders treatment. However, the application and the mechanism of EH against OV remain unexplored. Methods: A cell proliferation assay was first performed to validate the inhibitory effect of EH extracts on ovarian cancer cells. Network pharmacology was then employed to recognize potential bioactive compounds and target genes of EH. A Venn diagram was built to obtain the common targets of EH and OV and the protein-protein interaction (PPI) network of EH-OV correlated target genes was established by STRING. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment approaches were adopted to further reveal the molecular mechanism. Drug-compound-target-pathwaydisease network of EH in OV treatment was constructed. The expression of the predicted targets, cell cycle changes and cell apoptosis levels were analyzed to confirm the predicted mechanism of EH. Results: EH extracts suppressed the proliferation of human ovarian cancer cells in a concentration- and time-dependent manner. Ten bioactive compounds and 290 compound-associated targets were collected for EH-OV common targets exploration. A total of 74 EH-OV correlated genes were gathered and enrichment analysis identified phosphatidylinositide 3-kinases (PI3K)/AKT signaling pathway as the main machinery. Drug-compound-target-pathway-disease network indicated the polypharmacological effect of EH. Finally, EH extracts were confirmed to inhibit cell proliferation and induce cell apoptosis of ovarian cancer cells by inactivating PI3K/AKT signaling pathway. Conclusions: This study demonstrated the anti-ovarian cancer activity of whole EH extracts and indicated that EH should be considered as a potential therapeutic strategy for ovarian cancer. [ABSTRACT FROM AUTHOR]

Donafenib is an improved version of sorafenib in which deuterium is substituted into the drug's chemical structure, enhancing its stability and antitumor activity. Donafenib exhibits enhanced antitumor activity and better tolerance than sorafenib in preclinical and clinical studies. However, the specific mechanism of its effect on hepatocellular carcinoma has not been reported. Iron deposition is a cell death pattern caused by disturbances in iron metabolism. Apoptosis is a form of programmed cell death. They may interact with each other during cell death. This study mainly explores the potential mechanism of donafenib activating the p53 signaling pathway, inducing iron deposition, and enhancing cell apoptosis in hepatocellular carcinoma. Hepa1-6 and Huh7 cells were treated with various concentrations of donafenib. Scratch healing and pore migration tests were conducted. Analyze apoptosis through flow cytometry and TUNEL fluorescence labeling. RNA sequencing was conducted on both untreated and donafenib-treated Huh7 cells. The key proteins involved in ferroptosis (SLC7A11, GPX4) and apoptosis (caspase3, caspase8, Bax, Bcl-2, p53) were then evaluated using immunoblotting and immunohistochemical staining. Reactive oxygen species (ROS) levels in the cancer cells were measured. Donafenib treatment resulted in a dose-dependent decrease in the proliferation, migration, and invasion capabilities of cancer cells. There was an increase in apoptosis rates and ROS accumulation, and a reduction in tumor volume. The key proteins involved in ferroptosis and apoptosis underwent significant changes. Donafenib activates the p53 signaling pathway, induce ferroptosis, and enhance apoptosis, suggesting its potential as an effective therapeutic agent for HCC. [ABSTRACT FROM AUTHOR]

Background: Alternative splicing is essential for the physiological and pathological development of the inner ear. Disruptions in this process can result in both syndromic and non-syndromic forms of hearing loss. DHX38, a DEAH box RNA helicase, is integral to pre-mRNA splicing regulation and plays critical roles in development, cell differentiation, and stem cell maintenance. However, its specific role in inner ear development remains undefined. Here, we utilized a dhx38 knockout zebrafish model to monitor the ear morphology and elucidate a crucial role for DHX38 in the development of the zebrafish inner ear. Methods: Bright-field morphological analysis and in situ hybridization were performed to observe ear morphology changes. Immunofluorescence and semi-quantitative RT-PCR were employed to test apoptotic cells and abnormal splicing. Results: The dhx38-/- mutant zebrafish showed significant inner ear impairments, including decrescent otocysts, absent semicircular canal protrusion, and smaller otoliths. These structural abnormalities were accompanied by substantial DNA damage and p53-dependent apoptosis within the inner ear cells. Alternative splicing analysis showed that genes related to DNA damage repair and inner ear morphogenesis are abnormal in dhx38 knockout mutants. In summary, we suggest that dhx38 promotes cell survival during the inner ear development of zebrafish by ensuring the correct splicing of genes related to DNA damage repair. [ABSTRACT FROM AUTHOR]

Osteosarcoma (OS) is a malignant bone sarcoma arising from mesenchymal stem cells. The biological role of Acyl-CoA synthetase long-chain family member 4 (ACSL4), recently identified as an oncogene in numerous tumor types, remains largely unclear in OS. In this study, we investigated the expression of ACSL4 in OS tissues using immunohistochemistry staining (IHC) staining of a human tissue microarray and in OS cells by qPCR assay. Our findings revealed a significant up-regulation of ACSL4 in both OS tissues and cells. To further understand its biological effects, we conducted a series of loss-of-function experiments using ACSL4-depleted MNNG/HOS and U-2OS cell lines, focusing on OS cell proliferation, migration, and apoptosis in vitro. Our results demonstrated that ACSL4 knockdown remarkably suppressed OS cell proliferation, arrested cells in the G2 phase, induced cell apoptosis, and inhibited cell migration. Additionally, a subcutaneous xenograft mice model was established to validate the in vivo impact of ACSL4, revealing ACSL4 silencing impaired tumor growth in the OS xenograft mice. Additionally, we discovered that ACSL4 could regulate the phosphorylation level of Smad2 through cooperative interactions, and treatment with a TGF-β inhibitor weakened the promoting effects of ACSL4 overexpression. In short, ACSL4 regulated OS progression by modulating TGF-β/Smad2 signaling pathway. These findings underscore ACSL4 as a promising therapeutic target for OS patients and contribute novel insights into the pathogenesis of OS. [ABSTRACT FROM AUTHOR]

Intracerebral hemorrhage (ICH) is a common cerebrovascular disease characterized by a high incidence, disability rate, and mortality. Epigallocatechin gallate (EGCG), a key catechin compound found in green tea, has received increasing attention for its potential neuroprotective and therapeutic effects in neurological disorders. Studies have indicated that EGCG may influence various signaling pathways and molecular targets, including the inhibition of oxidative stress, reduction of inflammatory responses, suppression of cell apoptosis, regulation of cell survival, and enhancement of autophagy. Although the exact mechanism of action of EGCG is not fully understood, it has become a focal point of research in various disciplines due to its promising potential. This study aims to investigate the effects of EGCG on oxidative stress, iron deposition, and cell apoptosis in rats with ICH, as well as to uncover the underlying mechanisms. An ICH rat model was created to simulate cerebral hemorrhage, while an in vitro model utilizing primary cortical neurons was developed. The neurons were pre-treated with EGCG before being exposed to Erastin and RSL3 to induce iron death. The levels of oxidative stress, iron deposition, and cell apoptosis were evaluated in both models. In the ICH model, EGCG was discovered to enhance the activation of the Keap1/P62/Nrf2 signaling pathway in both in vivo and in vitro studies. Furthermore, EGCG significantly elevated the levels of GPX4 and XCT proteins, as well as the nuclear expression of Nrf2. It was noted that the Nrf2 inhibitor ML385 partially decreased the expression of these proteins. Through the activation of the Keap1/P62/Nrf2 pathway, EGCG inhibits inflammation, oxidative stress and iron deposition in rats with cerebral hemorrhage. EGCG inhibits oxidative stress, iron deposition and apoptosis in rats with ICH by activating Keap1/P62/Nrf2 pathway. [ABSTRACT FROM AUTHOR]

BACKGROUND: It has been found that vascular endothelial growth factor 165 and bone morphogenetic proteins interact with each other during hypoxiareoxygenation and are involved in the repair process of osteoblast injury by regulating the activation of intracellular signaling pathways. OBJECTIVE: To further investigate the relationship between vascular endothelial growth factor 165/bone morphogenetic protein and hypoxic-reoxygenated osteoblast injury. METHODS: Osteoblasts were selected and the hypoxic-reoxygenated injury model was established. Vascular endothelial growth factor 165 and bone morphogenetic protein expressions at mRNA and protein levels were detected by real-time PCR and western blot before and after modeling. After modeling, osteoblasts were given different concentrations of vascular endothelial growth factor 165 and bone morphogenetic protein 2 (10, 20, 40 ng/mL). Cell proliferation was detected by cell counting kit-8 method and apoptosis was detected by DAPI at 12, 24, 36, 48, and 72 hours after treatment. RESULTS AND CONCLUSION: Compared with before modeling, the mRNA and protein expressions of vascular endothelial growth factor 165 and bone morphogenetic protein 2 in osteoblasts after modeling were significantly decreased (P < 0.05). The proliferation rate of osteoblasts was significantly increased with the increase of vascular endothelial growth factor 165 concentration (P < 0.05), while the apoptosis rate of osteoblasts decreased significantly with the increase of vascular endothelial growth factor 165 concentration (P < 0.05). The proliferation rate of osteoblast was significantly increased with the increase of bone morphogenetic protein 2 concentration (P < 0.05), while the apoptosis rate of osteoblast decreased significantly with the increase of bone morphogenetic protein 2 concentration (P < 0.05). To conclude, vascular endothelial growth factor 165 and bone morphogenetic protein are lowly expressed in hypoxicreoxygenated osteoblast injury, and treatment with vascular endothelial growth factor 165 and bone morphogenetic protein can reduce the injury of hypoxicreoxygenated osteoblast in a concentration-dependent manner, suggesting that vascular endothelial growth factor 165 and bone morphogenetic protein have a significant protective effect against the injury of hypoxic-reoxygenated osteoblasts. [ABSTRACT FROM AUTHOR]

Objective To investigate the inhibitory effect and elucidate the molecular mechanism of 6-gingerol on human multiple myeloma cells. Methods The human multiple myeloma cell lines RPMI 8226 and ARH77 were cultured in vitro, followed by treatment with varying concentrations (50, 100, 200, 300, 400 μmol/L) of 6-gingerol. The inhibitory effect on cell proliferation was assessed using the CCK- 8 assay. Flow cytometry was employed to evaluate cell apoptosis and cycle distribution. Additionally, qRT-PCR and Western blotting techniques were utilized to analyze gene and protein expression levels. Results The proliferation of RPMI 8226 and ARH77 cells was dose- and time-dependently inhibited by 6-gingerol, leading to the induction of apoptosis with statistically significant differences (P < 0.05). Further mechanistic investigations revealed that treatment with 6-gingerol arrested RPMI 8226 cells in the G0/G1 phase, resulting in a significant increase in Bax levels and a decrease in Bcl-2 mRNA and c-Myc mRNA levels (P < 0.05). Additionally, it significantly upregulated the expression of Bax, Cleaved-PARP, Cleaved- caspase3, P53, and p-AKT proteins while down regulating the expression of Bcl-2 protein (P < 0.05). Conclusions The compound 6-Gingerol exhibits inhibitory effects on the proliferation and induction of apoptosis in MM cells, as well as cell cycle arrest at the G0/G1 phase. Its mechanism of action is likely associated with the suppression of the AKT signaling pathway, downregulation of Bcl-2 family protein expression, and inhibition of c-Myc expression. [ABSTRACT FROM AUTHOR]

Background: This study aims to improve the solubility and the toxicity of Bufonis venenum, and finally enhance the therapeutic outcomes of hepatocellular carcinoma (HCC). Methods: The cholesterol-free liposomes simultaneously encapsulate bufadienolides and indolealkylamines (Non-Cholesterol-Bufonis Venenum Extract-Liposome, Non-Chol-BVE-LP) was prepared by the thin-film evaporation technique. In vitro , the cytotoxicity, cell apoptosis study, cellular uptake and hemolysis studies were evaluated in HepG2 cells. In vivo , the biodistribution and anti-tumor activity studies were conducted in BALB/C mice with HepG2 cells. Results: The liposomes showed good size distribution, encapsulation efficiency drug loading capacity and slower drug release. Non-Chol-BVE-LP had higher cytotoxicity on HepG2 cells and induced more apoptosis on HepG2 Cells compared with BVE. In addition, the liposomes could accumulate in tumor by passive targeting, thus facilitating the anti-tumor effects. In vivo , Non-Chol-BVE-LP showed equivalent anti-tumor efficacy to the first-line anti-HCC drug sorafenib. Conclusion: The study provided new ideas for the development and clinical application of Bufonis venenum related formulation and offered new drug for the treatment of HCC. [ABSTRACT FROM AUTHOR]

Fabry disease, a lysosomal storage disease, is an uncommon X-linked recessive genetic disorder stemming from abnormalities in the alpha-galactosidase gene (GLA) that codes human alpha-Galactosidase A (α-Gal A). To date, over 800 GLA mutations have been found to cause Fabry disease (FD). Continued enhancement of the GLA mutation spectrum will contribute to a deeper recognition and underlying mechanisms of FD. In this study, a 27-year-old male proband exhibited a typical phenotype of Fabry disease. Subsequently, family screening for Fabry disease was conducted, and high-throughput sequencing was employed to identify the mutated gene. The three-level structure of the mutated protein was analyzed, and its subcellular localization and enzymatic activity were determined. Apoptosis was assessed in GLA mutant cell lines to confirm the functional effects. As a result, a new mutation, c.777_778del (p. Gly261Leufs*3), in the GLA gene was identified. The mutation caused a frameshift during translation and the premature appearance of a termination codon, which led to a partial deletion of the domain in C-terminal region and altered the protein’s tertiary structure. In vitro experiments revealed a significant reduction of the enzymatic activity in mutant cells. The expression was noticeably decreased at the mRNA and protein levels in mutant cell lines. Additionally, the subcellular localization of α-Gal A changed from a homogeneous distribution to punctate aggregation in the cytoplasm. GLA mutant cells exhibited significantly higher levels of apoptosis compared to wild-type cells. [ABSTRACT FROM AUTHOR]

Objective: To investigate influences of echinacoside (ECH) on myocardial cell apoptosis, inflammatory injury and mammalian target of rapamycin (mTOR)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in rats with coronary heart disease (CHD). Methods: A total of 12 SD rats were taken as control group (NC group), 48 CHD rat models were successfully constructed, and randomly divided into model (Model) group, ECH group (50 mg/kg), mTOR activator MHY1485 group (10 mg/kg), ECH+MHY1485 group (50 mg/kg ECH+10 mg/kg MHY1485). After 4 weeks, electrocardiogram was recorded through biological function experimental system. Kits were used to detect myocardial C-reactive protein (CRP) and IL-6 levels. Myocardial pathology and apoptosis were measured by HE and TUNEL staining. qRT-PCR was used to detect myocardial Bcl-2, Bax mRNA expressions. Western blot was used to detect myocardial mTOR/STAT3 pathway protein expression. Results: Compared with NC group, Model group rats had obvious necrosis, edema and inflammatory cell infiltration, ST segment deviation, IL-6, CRP levels, cardiomyocyte apoptosis rate, Bax mRNA expression, mTOR and p-STAT3/STAT3 protein levels were significantly increased (P< 0.05), Bcl-2 mRNA expression was significantly decreased (P<0.05). Compared with Model group, myocardial injury in ECH group was alleviated, ST segment shift, IL-6, CRP levels, cardiomyocyte apoptosis rate, Bax mRNA expression, mTOR and p-STAT3/STAT3 protein levels were were significantly decreased (P<0.05), Bcl-2 mRNA expression was significantly increased (P<0.05). The above indexes of MHY1485 group rats showed opposite trend. MHY1485 eliminates beneficial effects of ECH on CHD rats. Conclusion: ECH may reduce myocardial inflammatory injury and apoptosis in CHD rats by down-regulating mTOR/STAT3 pathway. [ABSTRACT FROM AUTHOR]

It is unknown what part Polyphyllin VII plays in HumanPapillomavirus (HPV)-positive cervical cancer, despite its recognized inhibitory effects on several types of cancer. The main focus of this work is to examine how Polyphyllin VII affects the proliferation and death of HPV-positive cervical cancer cells. To begin with, Hela and Caski cells were treated with varying doses of Polyphyllin VII. Subsequently, cell counting kit-8 (CCK-8) was used to evaluate the viability. Cell proliferation, cloning, cycle and apoptosis were analysed by flow cytometry and Western blot analysis was used to look at protein expression related to the phosphatidylinositol 3-kinase/Protein Kinase B (PI3K/Akt) pathway. The results showed that medium and high concentrations of Polyphyllin VII treatment of HPV-positive cells caused a notable decline in cell viability and crystal violet number, and prevented cells from transitioning from G1 phase to S phase. The rate of apoptosis also increased significantly, and there was a noticeable decline in the expression of p-PI3K/PI3K and p-Akt/Akt. Thus, it is concluded that medium and high concentrations of Polyphyllin VII inhibit the proliferation of HPV-positive cells and cause both apoptosis and cell cycle halt. Polyphyllin VII impedes the proliferation of HPV-positive cervical cancer cells through the PI3K/Akt pathway. [ABSTRACT FROM AUTHOR]

Background: The most common type of lung cancer is non-small cell lung cancer (NSCLC), accounting for 85% of all cases. Programmed cell death (PCD), an important regulatory mechanism for cell survival and homeostasis, has become increasingly prominent in cancer research in recent years. As such, exploring the role of PCD in NSCLC may help uncover new mechanisms for therapeutic targets. Methods: We utilized the GEO database and TCGA NSCLC gene data to screen for co-expressed genes. To delve deeper, single-cell sequencing combined with spatial transcriptomics was employed to study the intrinsic mechanisms of programmed cell death in cells and their interaction with the tumor microenvironment. Furthermore, Mendelian randomization was applied to screen for causally related genes. Prognostic models were constructed using various machine learning algorithms, and multi-cohort multi-omics analyses were conducted to screen for genes. In vitro experiments were then carried out to reveal the biological functions of the genes and their relationship with apoptosis. Results: Cells with high programmed cell death activity primarily activate pathways related to apoptosis, cell migration, and hypoxia, while also exhibiting strong interactions with smooth muscle cells in the tumor microenvironment. Based on a set of programmed cell death genes, the prognostic model NSCLCPCD demonstrates strong predictive capabilities. Moreover, laboratory experiments confirm that SLC7A5 promotes the proliferation of NSCLC cells, and the knockout of SLC7A5 significantly increases tumor cell apoptosis. Conclusions: Our data indicate that programmed cell death is predominantly associated with pathways related to apoptosis, tumor metastasis, and hypoxia. Additionally, it suggests that SLC7A5 is a significant risk indicator for the prognosis of non-small cell lung cancer (NSCLC) and may serve as an effective target for enhancing apoptosis in NSCLC tumor cells. [ABSTRACT FROM AUTHOR]