Search

Hedyotis diffusa Willd. (HDW), a traditional Chinese medicinal plant, exhibits a variety of pharmacological effects and has anticancer potential for a wide range of cancer types; Ferroptosis is a non-apoptosis-regulated cell death induced by iron accumulation and subsequent lipid peroxidation; and there is currently an increasing interest in the therapeutic role of ferroptosis in cancer. However, the effects of HDW on bladder cancer and its underlying molecular mechanisms remain largely unknown. In this study, a combination of in vivo and in vitro experiments, network pharmacology and data mining methods were used to investigate the effects of HDW on BLCA. The results showed that HDW exerted its anticancer activity by inducing ferroptosis in bladder cancer cells. Subsequently, we demonstrated for the first time that HDW induced ferroptosis in vitro and in vivo. To further explore the possible targets of HDW-induced ferroptosis in bladder cancer, we performed network pharmacological analyses, transcriptomic analyses, and single-cell analyses; through integrative analyses, we identified three key pivotal genes associated with iron death, CAV1, VEGFA, and JUN.Mechanistically, we showed that CAV1, VEGFA and JUN are key determinants of HDW-induced ferroptosis in BLCA. Knockdown of target genes altered the anticancer effects of HDW in 5637 and T24 cells. In conclusion, our data show for the first time that HDW exerts its anticancer effects on BLCA through CAV1, VEGFA and JUN gene-induced ferroptosis. This is expected to provide a promising compound for bladder cancer therapy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Diabetic retinopathy (DR) has been proven to be a leading cause of blindness. This study aimed to investigate the effect of Yes-associated protein 1 (YAP1) on the hypoxia-induced DR mice retinal microvascular endothelial cells (MRMECs) model. The hypoxia-induced DR MRMECs model was generated by treating in hypoxia circumstance (5 % CO 2 and 3 % O 2 ) for 48 h. This study constructed YAP1 overexpression and taurine-upregulated gene 1 (TUG1) silencing lentiviral vectors, both of which were used to infect the DR MRMECs model. Quantitative real-time PCR (qRT-PCR) was used to amplify the YAP1, TUG1, vascular endothelial growth factor A (VEGFA), and miR-144-3p gene. Western blot was used to identify the expression of YAP1 and VEGFA. The CCK-8 assay was used to evaluate proliferation and the flow cytometry assay was used to determine apoptosis of MRMECs. Cell migration and tube formation were also evaluated. The results showed that YAP1 overexpression and TUG1 silencing lentivirus were successfully constructed. YAP1 overexpression significantly promoted, but TUG1 silence inhibited cell proliferation and migration compared to DR MRMECs model (P<0.05). YAP1 markedly promoted TUG1/VEGFA and reduced miR-144-3p gene transcription compared to those of the DR MRMECs model (P<0.05). YAP1 overexpression and TUG1 silence demonstrated the opposite effects on VEGFA expression. YAP1 overexpression obviously promoted tube formation of MRMECs. In conclusion, overexpression of YAP1 promoted cell proliferation, cell migration, TUG1 and VEGFA expression, and reduced the transcription of the miR-144-3p gene in DR MRMECs. Overexpression of YAP1 aggravated the progress of DR in MRMECs by activating the TUG1/miR-144-3p/VEGFA signaling pathway., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The objective was to address the repercussion of adding titanium dioxide nanotubes (TiO 2 -nt) into high-viscosity conventional glass ionomer cement (GIC) on the biological properties of pre-odontoblastic cells (MDPC-23) challenged by lipopolysaccharides (LPS - 2 μg/mL). TiO 2 -nt was added to Ketac Molar EasyMix at 3, 5, 7 %, whereas unblended GIC served as control. Analyses included proliferation (n=6; 24, 48, 72 h), metabolism (MTT; n=6; 24, 48, 72 h); morphology laser microscopy (n=3; 24, 48, 72 h); proteome assessments IL-1β, IL-6, IL-10, VEGF, TNF-α (n=3; 12, 18 h); mRNA levels (RT-PCR) of Il-1β, Il-6, Il-10, VEGF, TNF-α (n=3; 12, 18 h) and DSPP (n=3; 24, 72, 120 h). Data analysis included Shapiro-Wilk, Levene, and generalized linear models (α=0.05). Results demonstrated that cell proliferation increased over time for all groups, and was not impacted by TiO 2 -nt (p>0.05). GIC groups displayed lower MTT values compared to cells cultured without GIC discs (p=0.019); disregarding the presence of TiO 2 . Remarkably, TiO 2 -nt reversed the effect of GIC, reducing the levels of selected biomarkers. LPS treatment modified the expression of the immune-inflammatory markers by MDPC-23 cells (p<0.0001). Morphological analysis did not reveal distinctions for any of the studied. TiO 2 -nt modulated immune-inflammatory and dentin marker expression by MDPC-23 cells cultured on conventional GIC discs, and did not affect cell morphology/viability, regardless LPS exposure. In conclusion, TiO 2 -nt may become a reliable clinical strategy to encourage pulp tissue repair., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kamila Rosamilia Kantovitz reports financial support was provided by State of Sao Paulo Research Foundation. Maria Davoli Meyer reports financial support was provided by State of Sao Paulo Research Foundation. Kamila Rosamilia Kantovitz reports administrative support and equipment, drugs, or supplies were provided by São Leopoldo Mandic College. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The hydroxyapatite orbital implantation is widely used to treat orbital malformation, but delayed postoperative angiogenesis can hinder conjunctival wound healing, potentially leading to implant exposure and prolapse. Low-intensity laser therapy (LLLT) is recognized for its ability to promote tissue regeneration, reduce inflammation, and alleviate pain. This study aims to explore the specific mechanism of miRNAs-VEGFA pathway regulation in early vascularization after orbital implant placement induced by LLLT. A hydroxyapatite orbital implant model was established and treated with LLLT. Vascular tissues surrounding the ocular prosthesis were extracted for high-throughput sequencing to identify differentially expressed miRNAs. miRNAs predicted to bind with VEGFA were selected for validation. GO and KEGG analyses were performed to reveal the functional enrichment of target genes regulated by these miRNAs. Dual luciferase assay, qRT-PCR, and Western blotting were used to verify the targeting relationship between miR-90 and VEGFA. The effects of miR-90 on rabbit microvascular endothelial cell function were assessed through CCK-8 assay, scratch test, and tube formation assay. High-throughput sequencing revealed 32 differentially expressed miRNAs, with 8 upregulated and 24 downregulated. miR-90 was predicted to have a high binding score and expression abundance with VEGFA and was confirmed to regulate VEGFA expression. In vitro functional tests showed that miR-90 inhibited rabbit microvascular endothelial cell proliferation, migration, and tube formation. This study is the first to demonstrate that LLLT regulates ocular prosthesis angiogenesis via the miR-90/VEGFA pathway, providing a new target for treating vascular-dependent diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Pathological neovascularization is a hallmark of many vision-threatening diseases. However, some patients exhibit poor responses to current anti-VEGF therapies due to resistance and limited efficacy. Recent studies have highlighted the roles of noncoding RNAs in various biological processes, paving the way for RNA-based therapeutics. In this study, we report a marked down-regulation of miR-205 under pathological conditions. miR-205 potently inhibits endothelial cell functions critical for pathological neovascularization, including proliferation, migration, and tube formation. Furthermore, miR-205 strengthens the endothelial barrier, thereby reducing vascular leakage. In mouse models of retinal and choroidal neovascularization, miR-205 administration effectively suppresses abnormal blood vessel formation and leakage. Mechanistically, miR-205 directly targets VEGFA and ANGPT2, which are key drivers of pathological neovascularization. To improve delivery, we successfully loaded miR-205 into exosomes derived from mesenchymal stem cells. This innovative approach avoids cytotoxicity while preserving therapeutic efficacy in both cellular and animal models. Collectively, our findings highlight miR-205 as a promising therapeutic for ocular neovascularization, with exosome delivery offering a novel and efficient strategy for treating vision-threatening vascular diseases., Competing Interests: Declarations. Ethics approval and consent to participate: All animal experiments were conducted according to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and approved by the Experimental Animal Center of Nanjing Medical University. Consent for publication: All authors agree to be published. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Over the past few years, biomaterial-based periodontal tissue engineering has gained popularity. An ideal biomaterial for treating periodontal defects is expected to stimulate periodontal-derived cells, allowing them to contribute most efficiently to tissue reconstruction. The present study focuses on evaluating the in vitro behavior of human periodontal ligament-derived stromal cells (hPDL-MSCs) when cultured on gelatin/Polycaprolactone prototype (GPP) and volume-stable collagen matrix (VSCM). Cells were cultured onto the GPP, VSCM, or tissue culture plate (TCP) for 3, 7, and 14 days. Cell morphology, adhesion, proliferation/viability, the gene expression of Collagen type I, alpha1 (COL1A1), Vascular endothelial growth factor A (VEGF-A), Periostin (POSTN), Cementum protein 1 (CEMP1), Cementum attachment protein (CAP), Interleukin 8 (IL-8) and Osteocalcin (OCN), and the levels of VEGF-A and IL-8 proteins were investigated. hPDL-MSCs attached to both biomaterials exhibited a different morphology compared to TCP. GPP exhibited stronger capabilities in enhancing cell viability and metabolic activity compared to VSCM. In most cases, the expression of all investigated genes, except POSTN, was stimulated by both materials, with GPP having a superior effect on COL1A1 and VEGF-A, and VSCM on OCN. The IL-8 protein production was slightly higher in cells grown on VSCM. GPP also exhibited the ability to absorb VEGF-A protein. The gene expression of POSTN was promoted by GPP and slightly suppressed by VSCM. In summary, our findings indicate that GPP electrospun nanofibers effectively promote the functional performance of PDLSCs in periodontal regeneration, particularly in the periodontal ligament and cementum compartment.

Cerebral aneurysms (CA) are critical conditions often associated with oxidative stress in vascular endothelial cells (VECs). The enzyme lactate dehydrogenase A (LDHA) plays a crucial role in glycolysis and lactate metabolism, processes implicated in the pathogenesis of aneurysms. Understanding these molecular mechanisms can inform the development of novel therapeutic targets. This study investigated the role of lactate metabolism and lactate-related genes, particularly LDHA and vascular endothelial growth factor A (VEGFA) genes, in VECs during oxidative stress. Using the GSE26969 dataset, we identified differential expression of lactate-related genes and performed functional enrichment analysis, revealing significant associations with glycolysis and lactate metabolic pathways. To induce oxidative stress, VECs were treated with H2O2, and the expression of LDHA and VEGFA was analyzed using quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting (WB) assays. Under oxygen-glucose deprivation/reperfusion (OGD/R) conditions, the effects of LDHA overexpression and VEGFA knockdown on cell viability and apoptosis were evaluated. Immunoprecipitation combined with western blotting was used to detect the lactylation status of LDHA following OGD/R stimulation and treatment with lactic acid (LA) and 2-deoxyglucose (2-DG). Our results indicated that oxidative stress modulates LDHA expression, glucose uptake, and lactate production, suggesting a metabolic shift towards glycolysis. LDHA overexpression improved cell survival and reduced apoptosis, while VEGFA knockdown had the opposite effect. Additionally, 2-DG treatment reduced LDHA lactylation and apoptosis. Our findings demonstrated that LDHA plays a critical role in the oxidative stress response of VECs, highlighting the potential therapeutic value of targeting glycolysis in CA. This study contributes to the understanding of metabolic adaptations in vascular pathologies and suggests new avenues for therapeutic intervention in CA management.

Angiogenesis is a significant character of lung adenocarcinoma (LUAD) and is an important reason leading to high mortality rates of LUAD patients. However, the molecular mechanisms of lncRNAs regulating the angiogenesis in LUAD have not been fully elucidated. Here we show lncRNA chromatin-associated RNA 10 (CAR10) was upregulated in the tumor tissue of patients with LUAD and enhanced tumor metastasis. Mechanistically, CAR10 could bind to Lactate Dehydrogenase A (LDHA) protein to regulate the phosphorylation and acetylation of LDHA and increase the dimerization of LDHA to promote its nuclear translocation, which increased the H3K79 methylation in Vascular Endothelial Growth Factor A (VEGFA) and Vascular Endothelial Growth Factor C (VEGFC) gene interval. CAR10 induced microvascular formation in vivo and in vitro by regulating LDHA-VEGFA/C axis. In addition, MYC and TP53 bonded to the promotor of CAR10 and reverse regulated its expression in LUAD cells. CAR10 regulates post-translational modification of LDHA and increases the H3K79 methylation of VEGFA/VEGFC to promote angiogenesis of LUAD, which is a potential therapeutic target for LUAD., Competing Interests: Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Introduction: The role of mast cells (MCs) in clear cell renal carcinoma (ccRCC) is unclear, and comprehensive single-cell studies of ccRCC MCs have not yet been performed., Methods: To investigate the heterogeneity and effects of MCs in ccRCC, we studied single-cell transcriptomes from four ccRCC patients, integrating both single-cell sequencing and bulk tissue sequencing data from online sequencing databases, followed by validation via spatial transcriptomics and multiplex immunohistochemistry (mIHC)., Results: We identified four MC signature genes (TPSB2, TPSAB1, CPA3, and HPGDS). MC density was significantly greater in ccRCC tissues than in normal tissues, but MC activation characteristics were not significantly different between ccRCC and normal tissues. Activated and resting MCs were defined as having high and low expression of MC receptors and mediators, respectively, whereas proliferating MCs had high expression of proliferation-related genes. The overall percentage of activated MCs in ccRCC tissues did not change significantly but shifted toward a more activated subpopulation (VEGFA + MCs), with a concomitant decrease in proliferative MCs (TNF + MCs) and resting MCs. An analysis of the ratio of TNF + /VEGFA + MCs in tumors revealed that MCs exerted antitumor effects on ccRCC. However, VEGFA + MC was produced in large quantities in ccRCC tissues and promoted tumor angiogenesis compared with adjacent normal tissues, which aroused our concern. In addition, MC signature genes were associated with a better prognosis in the KIRC patient cohort in the TCGA database, which is consistent with our findings. Furthermore, the highest level of IL1B expression was observed in macrophages in ccRCC samples, and spatial transcriptome analysis revealed the colocalization of VEGFA + MCs with IL1B + macrophages at the tumor-normal interface., Discussion: In conclusion, this study revealed increased MC density in ccRCC. Although the proportion of activated MCs was not significantly altered in ccRCC tissues compared with normal tissues, this finding highlights a shift in the MC phenotype from CTSG high MCs to more activated VEGFA+MCs, providing a potential therapeutic target for inhibiting ccRCC progression., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025 Song, Jiao, Qin, Zhang, Qin and Ma.)

Background: Cancer-targeted therapies are progressively pivotal in oncological care. Observational studies underscore the emergence of cancer therapy-related cardiovascular toxicity (CTR-CVT), impacting patient outcomes. We aimed to investigate the causal relationship between different types of cancer-targeted therapies and cardiovascular disease (CVD) outcomes through a two-sample Mendelian randomization (MR) study., Methods: This genome-wide association study was conducted using a two-sample Mendelian randomization framework. Genetic instruments for drug target gene expression were extracted from the eQTLGen consortium (31684 individuals, 37 cohorts). Genome-wide association study (GWAS) summary statistics for 19 cardiovascular diseases were derived from the FinnGen database. Primary analysis was carried out using the summary-data-based MR (SMR) method, with sensitivity analysis for validation. Colocalization analysis identifies shared causal variants between exposure eQTLs and CVD-associated single-nucleotide polymorphisms (SNPs)., Results: Among the 39 drug target genes, 8 were identified with detectable cis-eQTLs and were subsequently validated through positive control analysis for further investigation. In the SMR and sensitivity analyses, genetically proxied VEGFA inhibition showed significantly strong association with stroke (odds ratio [OR] = 1.17, 95% confidence interval [CI] = 1.09-1.26, p = 1.33 × 10 - 5 ). Additionally, the inhibition of FGFR1, FLT1, and MAP2K2 exhibited suggestive association with corresponding cardiovascular disease outcomes. Nevertheless, only VEGFA expression and stroke shared a causal variant (93.6%), whereas FGFR1, MAP2K2, and FLT1 did not share causal variants with corresponding cardiovascular diseases in the colocalization analysis., Conclusions: This genetic association study revealed evidence supporting the genetic association between the use of VEGFA inhibitors and increased stroke risk, highlighting the need for enhanced pharmacovigilance. These findings underscore the delicate balance between cardiovascular toxicity risk and the benefits of cancer-targeted therapy., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare that they have no competing interests., (© 2024. The Author(s).)

Triple-negative breast cancer (TNBC) remains a challenging subtype due to its aggressive nature and limited treatment options. This study investigated the potential synergistic effects of Korean mistletoe lectin ( Viscum album L. var. coloratum agglutinin, VCA) and cisplatin on MDA-MB-231 TNBC cells using both 2D and 3D culture models. In 2D cultures, the combination of VCA and cisplatin synergistically inhibited cell proliferation, induced apoptosis, and arrested the cell cycle at the G2/M phase. Also, the combination treatment significantly reduced cell migration and invasion. Gene expression analysis showed significant changes in specific genes related to apoptosis (Bax, Bcl-2), metastasis (MMP-2, MMP-9), and EMT (E-cadherin, N-cadherin). Three-dimensional spheroid models corroborated these findings, demonstrating enhanced cytotoxicity and reduced invasion with the combination treatment. Significantly, the 3D models exhibited differential drug sensitivity compared to 2D cultures, emphasizing the importance of utilizing physiologically relevant models in preclinical studies. The combination treatment also reduced the expression of angiogenesis-related factors VEGF-A and HIF-1α. This comprehensive study provides substantial evidence for the potential of VCA and cisplatin combination therapy in TNBC treatment and underscores the significance of integrating 2D and 3D models in preclinical cancer research.

Abdominal aortic aneurysm (AAA) is a severe cardiovascular disease (CVD) that is partly attributable to endothelial dysfunction, inflammatory response, and angiogenesis. G protein-coupled receptor 4 (GPR4), a proton-sensitive G protein-coupled receptor that is abundantly expressed in vascular endothelial cells, has been associated with numerous physiological functions. Nevertheless, its potential involvement in the development of AAA remains unexplored. In this study, we examined the impact of GPR4 deletion on the development of AAA in ApoE-deficient mice. The mice were categorized into four distinct groups: the ApoE-/- with saline group, the ApoE-/-GPR4-/- with saline group, the ApoE-/- with Ang II group, and the ApoE-/-GPR4-/- with Ang II group. AAA were induced in the ApoE-/- mice through the perfusion of angiotensin II (Ang II). Notably, GPR4 was substantially elevated in the AAA tissues from both human subjects and experimental mice. The deletion of GPR4 substantially decreased the formation of Ang II-induced AAA, damages to elastin, and the expression of aortic inflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α), as well as vascular endothelial growth factor A/vascular endothelial growth factor receptor 2 (VEGF-A/VEGF-R2), in ApoE-/- mice. Human aortic endothelial cells (HAECs) were transfected with lenti-viral GPR4 shRNA and subsequently stimulated with Ang II. Our findings indicate that the knockout of GPR4 attenuated Ang II-induced angiogenic tube formation in HAECs by decreasing the expression of VEGF-A and VEGF-R2. Furthermore, GPR4 knockout also hindered the activation of specificity protein-1 (SP-1) by reducing its expression and transcriptional activity. Notably, the overexpression of SP-1 reversed the inhibitory effects of GPR4 knockout on angiogenic tube formation and the expression of VEGF-A/VEGF-R2. This suggests that the protective effects of GPR4 knockout are achieved through the inhibition of SP-1. In summary, the absence of GPR4 impeded AAA formation, indicating that GPR4 could potentially serve as a therapeutic target for AAA., (© 2025 Wiley Periodicals LLC.)

Gliomas are common brain tumors. Despite extensive research, the 5-year survival rate of glioma remains low. Many studies have reported that circular RNAs (circRNAs) play a role in promoting the malignant progression of glioma; however, the role of circ&#95;0059914 in this process remains unclear. In this study, we aimed to investigate the function and underlying mechanism of circ&#95;0059914 in glioma. Western blotting and qRT-PCR were used to determine the levels of circ&#95;0059914, miR-1249, VEGFA, N-cadherin, vimentin, Snail, and EIF4A3. EDU and colony formation assays were conducted to evaluate cell proliferation. Transwell assays were used to explore cell migration and invasion and tube formation assays were used to analyze angiogenesis. RNA immunoprecipitation (RIP) and dual-luciferase reporter assays were used to explore the relationship between EIF4A3, circ&#95;0059914, miR-1249, and VEGFA. A xenograft tumor assay was performed to determine the role of circ&#95;0059914 in vivo. Circ&#95;0059914 expression was upregulated in gliomas. Knockdown of gliomal circ&#95;0059914 expression reduced the proliferation, migration, invasion, epithelial-mesenchymal transition (EMT), angiogenesis, and growth of glioma cells in vivo. Circ&#95;0059914 sponged miR-1249, and miR-1249 inhibition reversed the circ&#95;0059914 knockdown-mediated effects in glioma cells. VEGFA was found to be a target gene of miR1249; overexpression of VEGFA reversed the effect of miR-1249 up-regulation in glioma. Finally, EIF4A3 increased the expression of circ&#95;0059914. EIF4A3-induced circ&#95;0059914 expression plays a role in promoting glioma via the miR-1249/VEGFA axis., Competing Interests: Declarations. Ethics Approval and Consent Participate: The present experiment was approved (approval no. 2022PS780K) by the Medical Ethics Committee of Shengjing Hospital (Shenyang, China). Consent for Publication: Not applicable. Competing Interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Atherosclerosis (AS) is a vascular disease associated with endothelial damage, plaque formation, and retinal neovascularization (RNV), leading to visual impairment. Research indicates that vascular endothelial dysfunction, lipid deposition, and inflammatory responses contribute to the formation of plaques and atherosclerotic lesions. Among the common complications, studies have shown that RNV and the molecular mechanisms of AS hold significant clinical importance. In this study, we identified the overexpression of the gene heat shock protein 90 (HSP90) through transcriptome sequencing. Subsequent protein expression analysis and inhibition experiments in corresponding animal models confirmed the crucial role of HSP90 in the modulation of this disease. Research findings revealed an increase in the expression of HSP90, HMGA2, Snail family transcriptional repressor 2 gene (SNAI2), CXC chemokine receptor 4 (CXCR4), and vascular endothelial growth factor (VEGF) in atherosclerotic mouse tissues. Inhibition of HSP90 expression reduced vascular neovascularization and downregulated the expression of HMGA2 and VEGF. Given that HSP90 can promote HMGA2 expression, which, in turn, facilitates angiogenesis, we conducted lentiviral infection experiments on primary retinal endothelial cells obtained from atherosclerotic mice, confirming the regulatory role of HSP90 in modulating HMGA2 expression through the SNAI2/CXCR4 signaling pathway and its involvement in retinal endothelial neovascularization. In conclusion, our study highlights the significant regulatory role of HSP90 in AS-induced RNV, providing a new target for disease treatment. Furthermore, this research extensively explores the mechanism of HSP90 in regulating RNV and associated signaling pathways, offering novel insights and laying a solid foundation for future studies in this disease domain., (© 2024 Federation of European Biochemical Societies.)

This systematic review and meta-analysis aimed to identify the association between genetic polymorphisms and tendon and ligament injuries in adolescent and adult athletes of multiple competition sports. The PubMed, Web of Science, EBSCO, Cochrane Library, and MEDLINE databases were searched until July 7, 2023. Eligible articles included genetic studies on tendon and ligament injuries and comparisons between injured and non-injured athletes. This review included 31 articles, comprising 1,687 injury cases and 2,227 controls, from a meta-analysis of 12 articles. We identified 144 candidate gene polymorphisms (only single nucleotide polymorphisms were identified). The meta-analyses included vascular endothelial growth factor A (VEGFA) rs699947, collagen type I alpha 1 rs1800012, collagen type V alpha 1 rs12722, and matrix metalloproteinase 3 rs679620. The VEGFA rs699947 polymorphism showed a lower risk of injuries in athletes with the C allele ([C vs. A]: OR=0.80, 95% CI: 0.65-0.98, I 2 = 3.82%, p =0.03). The risk of these injuries were not affected by other polymorphisms. In conclusion, the VEGFA rs699947 polymorphism is associated with the risk of tendon and ligament injuries in athletes. This study provides insights into genetic variations that contribute to our understanding of the risk factors for such injuries in athletes., Competing Interests: There is no conflict of interest for any author., (Thieme. All rights reserved.)

Introduction: Hypoxia-inducible factor 1α [HIF1α] regulates gene expression, allowing the organism to respond to low oxygen levels. Meanwhile, astrocytes participate in inflammatory processes and are associated with neurotoxic chemicals that can increase stroke volume, contributing considerably to the devastating effects of a stroke., Objective: To evaluate whether Hif-1α ablation from the central nervous system is implicated in motor dysfunction and ischemic brain damage following stroke. Furthermore, to explore if Hif-1α ablation affects the therapeutic impact of NeuroD1 gene-based therapy., Methods: Endothelin-1 [ET-1] was injected to induce ischemic stroke in mice. Both wild-type and Hypoxia-inducible factor 1α conditional knockout [ Hif-1α CKO] mice were used. The effect of Hif-1α ablation was assessed by the neuron numbers, astrocyte activity, vascular endothelial growth factor [VEGF] expression, and behavioral tests. Moreover, western blot, ELISA, and RNA sequencing were used. Then, we used pAAV2/9-GfaABC1D-NeuroD1-P2A-EGFP-WPRE injection to examine the impact of NeuroD1 in Hif-1α CKO mice following ischemic stroke., Results: We found that following stroke, motor dysfunction significantly increased in Hif-1α CKO mice. Furthermore, elevation of apoptosis and activation in both microglia and astrocytes were observed, consequently up-regulating neuroinflammation. Meanwhile, Hif-1α ablation significantly decreased the efficiency of NeuroD1 gene-based therapy., Conclusion: Our findings demonstrate that Hif-1α ablation from the nervous system is implicated in ischemic stroke pathogenesis mainly by increasing neuron cell death and inducing astrocytes as well as decreasing the efficiency of NeuroD1. These data support the idea that manipulating HIF-1α is a viable therapeutic for ischemic stroke.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are among the most prevalent complications associated with cardiac surgery involving extracorporeal circulation (ECC), contributing to adverse outcomes and representing a significant impediment to successful cardiac surgical procedures. Vascular endothelial growth factor (VEGF) is implicated in the etiology of ALI/ARDS; however, its precise role remains a subject of debate due to the presence of somewhat contradictory findings in the literature, necessitating further investigation. To date, numerous studies have explored the role of VEGF in the pathophysiology of ALI/ARDS, with ongoing discussions regarding whether VEGF exerts a protective or detrimental effect. The genetic polymorphism of the VEGF gene is a significant factor in the development of ALI/ARDS. Research has indicated that the prevalence of the VEGF polymorphic gene is markedly higher in postoperative cardiac surgery patients who develop ALI/ARDS compared to the general population. Furthermore, the mortality rate among patients possessing the VEGF polymorphic gene is significantly elevated. Concurrently, it has been demonstrated that ARDS patients who are positive for the VEGF polymorphism exhibit a reduction in VEGF levels within alveolar lavage fluid, which correlates with an exacerbation of lung injury. The present paper provides a comprehensive review of the genetic polymorphisms of VEGF and their implications in the pathophysiological alterations observed in postoperative cardiac surgery patients with ALI/ARDS, thereby offering novel insights and evidence to further elucidate the mechanisms underlying ALI/ARDS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Vascular endothelial growth factor A 165 (VEGF-A 165 ) and VEGF receptor 2 (KDR) are important mediators of angiogenesis. We aimed to express the soluble KDR ligand-binding domain (sKDR1-3) and evaluate its interaction with the VEGF-A 165 receptor-binding domain (VEGFA 165 -RBD). sKDR1-3 DNA was designed and subcloned into pPinkα-HC plasmid. The cassette was transfected into the Pichia pink™ 4 genome by homologous recombination. We optimized the expression of sKDR1-3 under the induction of different methanol concentrations. VEGFA 165 -RBD was expressed in E. coli BL21 harboring pET28a( +)─VEGFA 165 -RBD vector under induction with IPTG with/without lactose. Interaction and biological activity of sKDR1-3 and VEGFA 165 -RBD were investigated by ELISA and anti-proliferation tests. sKDR1-3 migrated on SDS-PAGE gel as a 35-180 kDa protein due to glycosylation. The relative expression level of sKDR1-3 under 1% methanol was higher than 0.5% and 4% methanol induction. IPTG and cysteine were suitable for induction and refolding of VEGFA 165 -RBD. 25 ng sKDR1-3 and 20 ng VEGFA 165 -RBD showed strong binding. sKDR1-3 bound to VEGFA 165 -RBD and VEGF-A 165 with dissociation constants of 0.148 and 0.2 nM, respectively. 4-10 nM concentrations of sKDR1-3 inhibited the proliferation of HUVE cells induced by 5 nM VEGFA 165 -RBD. In consideration, sKDR1-3 in the nanomolar concentration range, is a promising anticancer drug to inhibit angiogenesis., Competing Interests: Declarations. Competing Interests: The authors have not disclosed any competing interests., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Pre-eclampsia is a placental disorder that affects 3-5% of all pregnancies and is a leading cause of maternal and fetal morbidity worldwide 1,2 . With no drug available to slow disease progression, engineering ionizable lipid nanoparticles (LNPs) for extrahepatic messenger RNA (mRNA) delivery to the placenta is an attractive therapeutic option for pre-eclampsia. Here we use high-throughput screening to evaluate a library of 98 LNP formulations in vivo and identify a placenta-tropic LNP (LNP 55) that mediates more than 100-fold greater mRNA delivery to the placenta in pregnant mice than a formulation based on the Food and Drug Administration-approved Onpattro LNP (DLin-MC3-DMA) 3 . We propose an endogenous targeting mechanism based on β 2 -glycoprotein I adsorption that enables LNP delivery to the placenta. In both inflammation- and hypoxia-induced models of pre-eclampsia, a single administration of LNP 55 encapsulating vascular endothelial growth factor (VEGF) mRNA resolves maternal hypertension until the end of gestation. In addition, with our VEGF mRNA LNP 55 therapeutic, we demonstrate improvements in fetal health and partially restore placental vasculature, the local and systemic immune landscape and serum levels of soluble Fms-like tyrosine kinase-1, a clinical biomarker of pre-eclampsia 1 . Together, these results demonstrate the potential of this mRNA LNP platform for treating placental disorders such as pre-eclampsia., Competing Interests: Competing interests: K.L.S., H.C.S., H.C.G. and M.J.M. have filed a patent application based on this work. M.J.M. is an inventor on a patent related to this work filed by the Trustees of the University of Pennsylvania (PCT/US20/56252). D.W. is an inventor on several patents related to this work filed by the Trustees of the University of Pennsylvania (11/990,646; 13/ 585,517; 13/839,023; 13/839,155; 14/456,302; 15/339,363; 16/299,202). The remaining authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Hypoxia is common in tumors and is associated with cancer progression and drug resistance, driven, at least in part, by genetic instability. Little is known on how hypoxia affects Translesion DNA Synthesis (TLS), in which error-prone DNA polymerases bypass lesions, thereby maintaining DNA continuity at the price of increased mutations. Here we show that under acute hypoxia, PCNA monoubiquitination, a key step in TLS, and expression of error-prone DNA polymerases increased under regulation of the HIF1α transcription factor. Knocking-down expression of DNA polymerase η, or using PCNA ubiquitination-resistant cells, inhibited genomic DNA replication specifically under hypoxia, and iPOND analysis revealed massive recruitment of TLS DNA polymerases to nascent DNA under hypoxia, uncovering a dramatic involvement of error-prone DNA polymerases in genomic replication. Of note, expression of TLS-polymerases correlates with VEGFA (primary HIF1α target) in a database of renal cell carcinoma, a cancer which accumulates HIF1α. Our results suggest that the tumor microenvironment can lead the cell to forgo, to some extent, the fast and accurate canonical DNA polymerases, for the more flexible and robust, but low-fidelity TLS DNA polymerases. This might endow cancer cells with resilience to overcome replication stress, and mutability to escape the immune system and chemotherapeutic drugs., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Aim: This study aims to examine the gene expression and DNA methylation patterns of angiogenic factors in the placentae of Indian women who underwent assisted reproductive technology (ART) procedures and their association with maternal one-carbon metabolites and birth outcome., Methods: Placental gene expression and DNA methylation of angiogenic factors ( VEGF , PlGF , FLT-1 , KDR ) in Indian women who underwent ART procedures ( n  = 64) and women who conceived naturally (Non-ART) ( n  = 93) was investigated using RT-qPCR and Epitect Methyl-II PCR assay kits. Maternal plasma one-carbon metabolites were assessed by CMIA technology., Result: Gene expression of FLT-1 and KDR was higher ( p  < 0.05) in the ART placentae. Placental global DNA methylation levels were higher ( p  < 0.01) and DNA methylation levels of VEGF promoter were lower ( p  < 0.05) in ART compared to non-ART women. Maternal plasma folate and vitamin B 12 levels were higher ( p  < 0.01) in the ART group. Gene expression of PlGF was negatively associated with maternal plasma folate ( p  < 0.05) whereas KDR was positively associated with maternal plasma homocysteine ( p  < 0.05). Gene expression of KDR was positively associated with chest circumference of the baby ( p  < 0.05)., Conclusion: Hypomethylation of VEGF and increased expression of FLT-1 and KDR was observed in the placentae of women who underwent ART procedure.

Retinal neovascularization (RNV) is a key pathological process in many blinding disorders. This study aims to investigate the potential mechanisms of heme oxygenase-1 (HO-1) on ferroptosis during RNV. Through bioinformatics analysis, differentially expressed ferroptosis-related genes were identified in the oxygen-induced retinopathy (OIR) mouse model. Ferroptosis was assessed in the OIR model and the human retinal microvascular endothelial cells (HRECs) with the treatment of H 2 O 2 . The mRNA and protein levels were measured through RT-qPCR and western blot. Lipid peroxidation was assessed through C11-BODIPY staining. HO-1 expression was knocked down by intravitreal injection with a self-complementary adeno-associated virus in the OIR model and small interfering RNA in HRECs. The pathological neovascular area and avascular area were assessed through immunofluorescent staining. The cellular functions of HRECs were evaluated with migration and tube formation assays. Our results demonstrated that HO-1 was significantly upregulated in the OIR model. 4-HNE upregulation and GPX4 downregulation were observed in the OIR model. The H 2 O 2 -induced oxidative stress resulted in lipid peroxidation, GPX4 downregulation, and mitochondrial morphology changes in HRECs. HO-1 knockdown induced GPX4 upregulation, and decreased lipid peroxidation in vitro and in vivo. Furthermore, HO-1 inhibition reduced pathological RNV in the OIR model and attenuated migration and tube formation in HRECs. Treatment with 6-OHDA restored the decrease of VEGFA, migration, and tube formation caused by HO-1 knockdown in HRECs. Overall, HO-1-mediated ferroptosis can regulate RNV through the COX2/VEGFA signal axis. These findings suggest that targeting HO-1 may serve as a promising approach for treating retinal neovascular diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Ovarian dysfunction is a major factor leading to female infertility. Understanding how to improve or reshape ovarian function has become an important entry point for preventing and treating female infertility caused by ovarian dysfunction. Here, plant-derived compounds are screened for in vitro activity upon ovarian organoids derived from feeder-free female germline stem cells. Tetrahydroxy stilbene glucoside (TSG) is found to promote the development of ovarian organoids. Single nucleus transcriptome sequencing and spatial transcriptome sequencing are used to establish a comprehensive spatiotemporal map to elucidate the role of TSG in ovarian organoid development, encompassing cell types and subtypes, transcription factors, pseudo-time sequence, and cell communication dynamics. This analysis indicates that TSG promotes ovarian organoid development through the vascular endothelial growth factor A-Eph receptor B2 ligand-receptor pair between granulosa cells and oocytes. This study has enhanced the understanding of the mechanisms of ovarian organoid development, establishes a technical platform for screening compounds for treating infertility and related diseases, and lays a foundation for clinically applying plant-derived compounds., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Nowadays, the use of monoclonal antibodies to target angiogenic signalling pathways is common, but, unfortunately, the clinical activity of these agents is limited. Thus, the development of approaches targeting multiple pathways for anti-angiogenic effect will lead to increase the clinical benefit. For this purpose, oleuropein, hesperidin, piperine, proanthocyanidins and retinoic acid, which have previously been proven to be bioactive components, anti-angiogenic performances were experimentally tested in retinal pigment epithelial cells. Bioactive ingredients were applied to retinal pigment epithelial cells at varying doses for 48 h, and then IC 50 doses were calculated by MTT analysis. VEGFA and VEGFR1 protein levels were measured by ELISA analysis. Using the RT-PCR technique, the mRNA expression levels of EGF, EGFR, PDGF, PDGFR-β and HIF1A were analysed. Among all bioactive compounds, the bevacizumab group showed the best anti-VEGF activity, followed by the proanthocyanidins and piperine groups. Piperine group showed EGF, PDGF and HIF1A expressions; proanthocyanidins, on the other hand, reduced EGFR, PDGF and HIF1A expressions. Retinoic acid showed an angiogenic effect by increasing VEGFA protein levels and EGF and PDGFR-β expressions. Although hesperidin increased VEGFA protein levels, it decreased EGF, PDGF, PFGFR-β and HIF1A expression levels. Among the bioactive components stated in previous studies to have anti-angiogenic properties, only proanthocyanidins and piperin had anti-VEGF properties. Considering that angiogenesis does not proceed only through VEGF, this study investigated and reported for the first time the effects of relevant bioactive components on angiogenesis through different mechanisms in retinal pigment epithelial cells., (© 2025 The Author(s). Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)

Apigenin, a dietary flavonoid, has gained increasing attention for its potential therapeutic applications in radiation protection and radiosensitization. Ionizing radiation (IR) can harm healthy cells, but as radiotherapy remains crucial in cancer treatment. Owing to the remarkable application of radiotherapy in the treatment of cancers, it is vital to protect healthy cells from radiation hazards while increasing the sensitivity of cancer cells to radiation. This article reviews the current understanding of apigenin's radioprotective and radiosensitive properties with a focuses on the involved signaling pathways and key molecular targets. When exposed to irradiation, apigenin reduces inflammation via cyclooxygenase-2 inhibition and modulates proapoptotic and antiapoptotic biomarkers. Apigenin's radical scavenging abilities and antioxidant enhancement mitigate oxidative DNA damage. It inhibits radiation-induced mammalian target of rapamycin activation, vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP), and STAT3 expression, while promoting AMPK, autophagy, and apoptosis, suggesting potential in cancer prevention. As a radiosensitizer, apigenin inhibits tumor growth by inducing apoptosis, suppressing VEGF-C, tumor necrosis factor alpha, and STAT3, reducing MMP-2/9 activity, and inhibiting cancer cell glucose uptake. Cellular and animal studies support apigenin's radioprotective and anticancer potential, making it a potential candidate for further research. Investigation into apigenin's therapeutic efficacy in diverse cancer types and radiation damage is essential., (© 2024 International Union of Biochemistry and Molecular Biology.)

Background/aim: Glioblastoma is the most malignant brain tumor, and despite advances in treatment, survival rates are still dismal. Therefore, a comprehensive understanding of the underlying molecular mechanisms of glioblastoma is needed. This study suggests potential therapeutic targets in glioblastoma that may provide new therapeutic insights., Materials and Methods: To identify hub genes in glioblastoma, three datasets were selected from the GEO database. After screening DEGs using GEO2R, GO and KEGG analyses were performed using DAVID. The PPI network was visualized using Cytoscape and 7 hub genes were extracted. The prognostic potential of 7 hub genes was investigated using the Gliovis and GEPIA2 databases., Results: In total, 176 up-regulated and 263 down-regulated genes were identified. From the PPI network, 7 hub genes were identified including CAMK2A, DLG4, SNAP25, SYT1, MYC, FN1, and VEGFA. Out of the 7 hub genes identified, FN1 and VEGFA have been associated with a poor prognosis in glioblastoma based on the survival analysis., Conclusion: This study suggests that high levels of FN1 and VEGFA expression are associated with a poor prognosis in glioblastoma and that both genes are promising targets for glioblastoma therapy. Bioinformatics analysis of DEGs revealed putative targets that might reveal the molecular mechanisms underlying glioblastoma., (Copyright © 2025, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Tumor-derived exosomes are highly correlated with tumor progression and angiogenesis. This study was designed to probe the role of tumor-derived exosomal miR-1247-3p in mediating the angiogenesis in bladder cancer. Exosomes isolation from the culture medium of normal or bladder cancer cell lines was performed using a differential centrifugation method. miR-1247-3p expression in exosomes and cells was detected by quantitative real-time PCR (qRT-PCR). The effect of exosomes on the angiogenesis of human umbilical vein endothelial cells (HUVECs) was assessed using cell counting kit-8 (CCK-8), transwell and tube formation assays. The interaction between miR-1247-3p and forkhead box protein O1 (FOXO1) was studied using luciferase reporter and RNA pull down assays. Exosomes were successfully isolated from T24, UM-UC-3, and SV-HUC-1 cells, as confirmed by corresponding identifications. Functional experiments revealed that exosomes derived from T24 and UM-UC-3 cells significantly enhanced the abilities of proliferation, migration, angiogenesis, and vascular endothelial-derived growth factor (VEGF) secretion in HUVECs. miR-1247-3p was highly expressed in exosomes derived from T24 and UM-UC-3 cells, and exosomes derived from miR-1247-3p inhibitor-transfected cells reduced HUVEC viability, migration, tube formation, and VEGF level. FOXO1 was confirmed as a direct target of miR-1247-3p. Rescue assays suggested that the effect of miR-1247-3p inhibition on the viability, migration, and angiogenesis of HUVECs was partly abrogated by the knockdown of FOXO1. Our data suggest that miR-1247-3p is up-regulated in tumor-derived exosomes, thereby inhibiting FOXO1 expression and facilitating angiogenesis in bladder cancer.

Hepatocellular carcinoma (HCC) remains a lethal malignancy with limited treatment options. Recent discoveries have highlighted the pivotal role of miRNAs in HCC progression. We previously reported that the expression of miR-200b-3p was decreased in HCC cells and exosomal miR-200b-3p from hepatocytes inhibited angiogenesis by suppressing the expression of the endothelial transcription factor ERG (erythroblast transformation-specific (ETS)-related gene), leading to the hypothesis that the delivery of this miRNA may inhibit angiogenesis and suppress HCC growth in vivo. Here, we tested this hypothesis by using human HCC inoculation models. First, we transfected the human HepG2 HCC cells and established a stable cell line that overexpressed a high level of miR-200b-3p. When miR-200b-3p-overexpressing cells were injected into severe combined immunedeficiency (SCID)-beige mice, tumor growth was significantly reduced compared to tumors of control cells, with a reduction in the expression of ERG and vascular endothelial growth factor (VEGF) and subsequent angiogenesis. Intra-tumoral injection of exosomes containing high levels of miR-200b-3p also reduced the growth of parental HepG2 tumors with reduced ERG and VEGF expression and angiogenesis. These results validate the inhibitory role of miR-200b-3p in tumor angiogenesis, thereby suppressing HCC tumor growth, and provide a novel insight into its potential therapeutic application., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Diabetic foot ulcer (DFU) is a common but devastating complication of diabetes mellitus and might ultimately lead to amputation. Elucidating the regulatory mechanism of wound healing in DFU is quite important for developing DFU management strategies. Here, we show, mecenchymal stem cell (MSC)-derived exosomes promoted the proliferation, migration and angiogenesis of high glucose-treated endothelial cells and reduced cell apoptosis. These effects were further enhanced by MSC-derived exosomes carrying circMYO9B overexpression. Mechanistically, circMYO9B promoted the translocation of hnRNPU from nucleus to cytoplasm and consequently destabilized CBL, thereby reducing the ubiquitination and degradation of KDM1A to promote VEGFA expression in endothelial cells. MSC-derived exosomes carrying circMYO9B promotes angiogenesis and thus accelerates diabetic wound healing through regulating the hnRNPU/CBL/KDM1A/VEGFA axis, indicating potential therapeutic targets and strategies for DFU treatment., Competing Interests: Competing interests: The authors declare no competing interests. Ethics approval and consent to participate: All donors in this study were informed and provided written consent, and our study was approved by the Ethics Committee of the Third Xiangya Hospital Central South University. Animal procedures were approved by the Animal Care and Use Committee of the Third Xiangya Hospital Central South University., (© 2024. The Author(s).)

Idiopathic pulmonary fibrosis (IPF) is a chronic pulmonary disease with unknown pathogenesis and poor prognosis. PANoptosis, a newly identified form of inflammatory programmed cell death, has been implicated in various inflammatory lung diseases. This study aimed to identify differentially expressed PANoptosis-related genes (PRDEGs) associated with immune infiltration and prognosis in IPF, while also establishing a novel prognostic prediction model. A total of 63 PRDEGs were identified from GSE110147 dataset, with 31 exhibiting consistent expression trends in GSE213001. Enrichment analysis indicated that the majority of these PRDEGs were enriched in inflammatory and immune-related pathways. Three key PRDEGs-NLRP3, ATM, and VEGFA-were selected through univariate and multivariable Cox regression analyses. The prognostic prediction model developed from these key PRDEGs demonstrated robust predictive performance. Furthermore, the expression of most PRDEGs was positively correlated with pro-inflammatory immune cells, including macrophages, neutrophils, and CD4 + T cells. Validation of the expression levels of these key PRDEGs was conducted in fibrotic mouse lung tissue. This study suggests that PANoptosis plays a role in IPF, potentially linked to the infiltration of pro-inflammatory immune cells, and may influence disease progression through the regulation of inflammatory immune signaling pathway. A new prognostic prediction model for IPF based on PRDEGs was successfully developed., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study investigated the role of C-X-C motif chemokine receptor 6 (CXCR6) in colorectal cancer (CRC). It was found that lower CXCR6 expression is correlates with poorer prognostic outcomes, suggesting that CXCR6 may inhibit tumor progression and thus improve patient outcomes. Silencing CXCR6 in CRC cell lines SW620 and CT-26 resulted in significantly enhanced migration and invasion, as demonstrated by wound healing and transwell assays. Further analysis revealed that CXCR6 activity is associated with activation of the VEGFA/PI3K/AKT/mTOR signaling pathway. Inhibition of this pathway through VEGFA siRNA and pathway-specific inhibitors reversed the effects of CXCR6 silencing on cell migration and invasion. Moreover, xenograft experiments showed that silencing CXCR6 led to increased tumor growth and upregulated proteins associated with the extracellular matrix and epithelial-mesenchymal transition. These findings were validated through immunohistochemical, immunofluorescence, and Western blot analyses. This study highlights CXCR6's critical role in CRC and its potential as a therapeutic target to manage cancer progression., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: The simultaneous differentiation of human pluripotent stem cells (hPSCs) into both endodermal and mesodermal lineages is crucial for developing complex, vascularized tissues, yet poses significant challenges. This study explores a method for co-differentiation of mesoderm and endoderm, and their subsequent differentiation into pancreatic progenitors (PP) with endothelial cells (EC)., Methods: Two hPSC lines were utilized. By manipulating WNT signaling, we optimized co-differentiation protocols of mesoderm and endoderm through adjusting the concentrations of CHIR99021 and mTeSR1. Subsequently, mesoderm and endoderm were differentiated into vascularized pancreatic progenitors (vPP) by adding VEGFA. The differentiation characteristics and potential of vPPs were analyzed via transcriptome sequencing and functional assays., Results: A low-dose CHIR99021 in combination with mTeSR1 yielded approximately 30% mesodermal and 70% endodermal cells. Introduction of VEGFA significantly enhanced EC differentiation without compromising PP formation, increasing the EC proportion to 13.9%. Transcriptomic analyses confirmed the effectiveness of our protocol, showing up-regulation of mesodermal and endothelial markers, alongside enhanced metabolic pathways. Functional assays demonstrated that vPPs could efficiently differentiate into insulin-producing β-cells, as evidenced by increased expression of β-cell markers and insulin secretion., Conclusion: Our findings provide a robust method for generating vPPs, which holds significant promise for regenerative medicine applications, particularly in diabetes treatment., Competing Interests: Declarations. Ethical approval and consent to participate: This study does not involve human participants or animals. Use of human pluripotent cell lines (UC and H1) was approved by the ethics committee of Shenzhen Hospital, Beijing University of Chinese Medicine in March 2021 (SZLDH2021LSYA-010). Consent for publication: Not applicable. Artificial intelligence: The authors declare that they have not use AI-generated work in this manuscript. Competing interests: The authors have declared no competing interests., (© 2024. The Author(s).)

Background: Mesenchymal stem cells may have neuroprotective and tissue regenerative capabilities and the potential to rescue retinal degeneration in chorioretinal diseases including myopic chorioretinal atrophy. Transplantation of human (allogeneic) adipose tissue-derived mesenchymal stem cell (adMSC) suspensions has been clinically conducted to treat retinal degenerative diseases. However, serious side effects including proliferative vitreoretinopathy and epiretinal membrane formation have been reported. PharmaBio Corporation fabricated novel adMSC sheets with a Bruch's membrane-like structure using our original method, potentially overcoming these problems. We evaluated the characteristics of newly developed adMSC sheets named PAL-222 and assessed their safety and efficacy in rats with congenital retinal degeneration (RCS rats) to obtain the proof-of-concept for the first-in-human clinical trial for myopic chorioretinal atrophy., Methods: We measured the viability of cells obtained from PAL-222, examined cell surface antigens by flow cytometry, measured the vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) secretory ability, examined the expression of types I and IV collagen and elastin by immunostaining. We performed a transwell in vitro migration assay to evaluate durability and similarity to retinal pigment epithelium (RPE) and checked in vitro tumorigenicity. In an in vivo experiment, we transplanted PAL-222 into the subretinal space of RCS rats and evaluated the safety and efficacy., Results: Viability of cells obtained from PAL-222 was 88.1%. The rate of positive markers such as CD90, CD73, CD105 and CD44 exceeded 90%; that of the negative markers such as CD34, CD11b, CD19, CD45 and HLA-DR was less than 2%. PAL-222 secreted significant amounts of VEGF and PEDF and expressed types I and IV collagen and elastin. The migration assay showed that PAL-222 preserved the sheet structure without cell migration. No chromosomal aberration or colony formation was observed in in vitro tumorigenicity tests. PAL-222 transplantation suppressed the progression of retinal degeneration by preserving the outer nuclear layer without negative changes in RCS rats, suggesting a retinoprotective effect., Conclusions: We confirmed the efficacy and safety of PAL-222 and are currently conducting a clinical trial to treat myopic chorioretinal atrophy. Transplantation of these novel adMSC sheets may be a promising therapy for myopic chorioretinal atrophy., Trial Registration: ClinicalTrials.gov, Identifier: NCT05658237. URL: https://classic., Clinicaltrials: gov/ct2/show/NCT05658237 ., Competing Interests: Declarations. Ethics approval and consent to participate: All study animals were treated according to the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research. The Animal Experiment Committee of Shin Nippon Biomedical Laboratories, Ltd., (2438 Miyanouracho, Kagoshima City, Kagoshima Prefecture) approved the study in advance ((1) Title of the approved project; Single Subretinal Dose Retinal Protection Efficacy Study for PAL-222 in RCS Rats, (2) Name of the institutional approval committee or unit; The Animal Experiment Committee of Shin Nippon Biomedical Laboratories, Ltd., (3) Approval number; SBL469-010, (4) Date of approval; 5/23/2022). The testing facility was accredited by AAALAC International. Human cells were managed according to the tenets of the Declaration of Helsinki. This study does not report on or involve the use of any human data. Consent for publication: Not applicable. Competing interests: NN and TY are advisors and shareholders of PharmaBio Corporation. Funders played a role in the conduct of the study, collection of data, management of the study, analysis of data, interpretation of data, or preparation of the manuscript., (© 2024. The Author(s).)

Background: Mitochondrial DNA (mtDNA) pathogenic variants have been reported in several solid tumors including ovarian cancer (OC), the most lethal gynecologic malignancy, and raised interest as they potentially induce mitochondrial dysfunction and rewiring of cellular metabolism. Despite advances in recent years, functional characterization of mtDNA variants in cancer and their possible modulation of drug response remain largely uncharted., Methods: Here, we characterized mtDNA variants in OC patient derived xenografts (PDX) and investigated their impact on cancer cells at multiple levels., Results: Genetic analysis revealed that mtDNA variants predicted as pathogenic, mainly involving complex I and IV genes, were present in all but one PDX (n = 20) at different levels of heteroplasmy, including 7 PDXs with homoplasmic variants. Functional analyses demonstrated that pathogenic mtDNA variants impacted on respiratory complexes activity and subunits abundance as well as on mitochondrial morphology. Moreover, PDX cells bearing homoplasmic mtDNA variants behaved as glucose-addicted and could barely survive glucose starvation in vitro. RNA-seq analysis indicated that mtDNA mutated (heteroplasmy > 50%) PDXs were endowed with upregulated glycolysis and other pathways connected with cancer metabolism. These findings led us to investigate whether pathogenic mtDNA variants correlated with response to anti-VEGF therapy, since the latter was shown to reduce glucose availability in tumors. Strikingly, PDXs bearing homoplasmic pathogenic mtDNA variants associated with improved survival upon anti-VEGF treatment in mice, compared with mtDNA wild type or low heteroplasmy PDXs., Conclusions: These results hint at mtDNA variants as potential biomarkers of response to antiangiogenic drugs., Competing Interests: Declarations. Ethics approval and consent to participate: All procedures involving animals and their care adhere to institutional guidelines that comply with national and international laws and policies (EEC Council Directive 2010/63/EU, OJ L 276, 20.10.2010). The animal study was reviewed and approved by the Italian Ministry of Health (Authorization n. 617/2016-PR), in accordance with the Declaration of Helsinki. Consent for publication: Not applicable. Competing interests: The authors declares that they have no competing interests., (© 2024. The Author(s).)

When a lower limb is injured, the most delicate organs that are at risk of harm are the lungs. Among the flavonoids, quercetin is a significant component that is found in apples and onions in the highest proportions. Numerous biological actions, including as anti-inflammatory, antioxidant, and anti-cancer properties, have been linked to quercetin. To investigate the impact of quercetin on lung injury induced by skeletal muscle ischemia reperfusion (IR) injury. Three equal groups of twenty-four adult rats were used: control, Ischemia-reperfusion (IR) group and IR group treated with quercetin. Rats in (IR) group were exposed to ischemia by ligation of femoral artery for 2h then after removal of the clamp, reperfusion was estabilished for another 24h. IR group treated with quercetin, rats were underwent hind limb IR as described in group II then were given quercetin that was administered at a dose of 20mg/kg intraperitoneally. Measurement of cytokines in serum, MDA in tissue homogenate and VEGF in serum and tissue homogrnate in addition to mRNA expression level and detection of protein level of both sirtuin-1(SIRT1) and NF-κB were assessed at the end of experiment. Histological and immunohistochemical assessment of the lungs were also carried. IR group showed notable rise of inflammatory cytokines such as IL-1β, IL-6 and TNF-α in addition to high level of VEGF and MDA in IR group when compared to the IR group treated with quercetin. Also, gene expression and protein level of SIRT1 were reduced while NF-κB mRNA expression and protein level were significantly upregulated in IR group compared to IR group treated with quercetin. Histologically, IR group indicated marked histological alterations of lung tissue. Also, IR showed strong brownish cytoplasmic immunostaining for iNOS and abundance of Ki67-positive cells. These alterations were significantly reversed in IR group treated with quercetin. Biochemical and immunohistochemical findings of this study demonstrate that quercetin administration have protective effects against lung injury induced by lower limb IR., Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Coronary artery disease (CAD) is a life-threatening condition caused by the chronic gradual narrowing of the lumen of the blood vessels of the heart by atherosclerotic plaque with a strong genetic component. The aim of our study was to investigate the association between the VEGFA polymorphism rs2010963 and myocardial infarction in patients with type 2 diabetes, as well as the expression of VEGFA. A total of 1589 unrelated Caucasians with T2DM lasting longer than 10 years were divided into two groups: case group subjects with MI (484) and a control group without a history of CAD (1105). A total of 25 endarterectomy sequesters were immunohistochemically stained to assess VEGFA expression. The rs2010963 polymorphism of the VEGFA gene was genotyped using a KBioscience Ltd. competitive allele-specific fluorescence-based PCR (KASPar) assay. The C allele was significantly more common in the case group according to the dominant model of inheritance (CC + CG vs. GG) (OR: 1.32; 95% CI: 1.05-1.66; p = 0.0197). A statistically significantly higher numerical areal density of VEGFA-positive cells was found in subjects with the C allele (CC + CG genotypes) in comparison to the GG genotype (117 ± 35/mm 2 vs. 58 ± 21/mm 2 ; p < 0.001). To conclude, the rs2010963 polymorphism is a potential genetic risk factor for myocardial infarction in Slovenian patients with T2DM.

Hepatocellular carcinoma (HCC) is a highly aggressive malignant tumor with a poor prognosis. This research aimed to investigate the role of F13B in HCC and its underlying mechanisms. Through comprehensive bioinformatics analysis of the GSE120123 and The Cancer Genome Atlas (TCGA)-Liver hepatocellular carcinoma (LIHC) datasets, we identified 220 overlapping prognosis-related genes. Eight key genes, including the previously unreported CCDC170 and F13B in HCC, were identified through Least Absolute Shrinkage and Selection Operator (LASSO)-Cox regression analysis. F13B emerged as a significant prognostic factor in HCC, warranting further investigation in subsequent analyses. In vitro experiments showed that F13B expression was notably reduced in HCC cell lines and tissues, particularly in Huh-7 and SMMC-7721 cells. Overexpression of F13B inhibited cell invasion, migration, and proliferation, while its knockdown produced the opposite effect. A lactate dehydrogenase (LDH) activity assay in human umbilical vein endothelial cells (HUVECs) demonstrated that F13B overexpression reduced vascular endothelial growth factor (VEGF)-induced cytotoxicity, whereas knockdown increased it. Further analysis revealed that F13B negatively regulates VEGFA expression, affecting HUVEC proliferation. In HUVECs, F13B overexpression reversed VEGF-induced upregulation of key angiogenesis markers, including phospho-VEGF receptor 2 (p-VEGFR2), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), as well as AKT/mTOR signaling proteins, phospho-Akt (p-AKT), and phospho-mTOR (p-mTOR). Additionally, F13B negatively regulated VEGFA and hypoxia-inducible factor 1 A (HIF1A) under hypoxic conditions, counteracting the hypoxia-induced increase in cell viability. These findings suggest that F13B regulates angiogenesis through the HIF-1α/VEGF pathway and plays a crucial role in HCC progression. Our results highlight the potential of F13B as a therapeutic target in HCC, providing novel insights into the molecular mechanisms of HCC and its prognostic significance.

Leptin, an immune-regulating protein, enhances the maturation of dendritic cells (DCs). We previously demonstrated that leptin and lipopolysaccharide (LPS) promote the expression of co-stimulatory molecules on the surface of DCs. Leptin/LPS-treated DCs increased T cell responses against 4T1 breast cancer in mice. Therefore, in the present study, we investigate the effects of a DC vaccine treated with leptin and LPS on the genes involved in tumor metastasis, angiogenesis, and related cytokines in a mouse model of breast cancer. Tumor induction was achieved through subcutaneous injection of 4T1 cells into syngeneic mice. On days 12 and 19, the mouse groups received the DC vaccine treated with leptin and a combination of leptin and LPS. After sacrificing the mice on day 26, the levels of IL-6 and IL-33 in the serum were assayed using the ELISA technique, and the expression levels of the VEGF, CCL2, MMP9, and CCL5 genes in the tumors were measured by Real-Time PCR. Compared to untreated tumor-bearing mice, the leptin-treated mature DC (mDC) group exhibited a significant reduction in the expression of MMP9 (0.33-fold, p = 0.01) and CCL5 (0.81-fold, p = 0.02). The leptin-LPS-treated mDC group showed decreased expression of genes involved in metastasis and tumor growth, including VEGF (0.72-fold, p = 0.03), MMP9 (0.26-fold, p = 0.001), and CCL5 (0.3-fold, p = 0.006), indicating more efficient prevention of metastasis. The CCL2 gene expression levels in both treatment groups showed a slight decreasing trend, but these changes were not statistically significant. The leptin-treated mDC group reduced IL-6 production by approximately 16% (p = 0.02), while treatment with the leptin-LPS-treated mDC significantly decreased IL-6 production by approximately 22% (p = 0.01) and increased IL-33 production by approximately 42% (p = 0.03). The findings of the present study indicate that the leptin-LPS-treated mDC vaccine group reduced the expression of genes and cytokines involved in metastasis and angiogenesis, demonstrating greater efficacy compared to the leptin-treated mDC vaccine group., Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Curcumin, found in turmeric rhizomes (Curcuma longa L.), has been widely studied for its potential health benefits, including anti-inflammatory, antioxidant, and wound-healing properties. However, due to its low bioavailability and unfavorable pharmacokinetics, analogous compounds have been developed to obtain better biopharmaceutical characteristics and enhanced biological effects. In this study, we evaluated the activity of curcumin and three of its synthetic analogues (DMAD, DMAM, and RI75) on the viability and differentiation of a pre-osteoblastic cell line (MC3T3-E1). We also assessed the expression of key genes involved in tissue regeneration: vascular endothelial growth factor (vegf), stromal-derived growth factor 1 (SDF-1/CXCL12), and runt-related transcription factor 2 (runx2). The cells were treated with curcumin and the three analogues at concentrations of 10, 30, or 50 μM. All tested analogues and curcumin exhibited moderate to no cell toxicity compared to the cells treated under standard conditions across all concentrations after 24, 48, and 72 hours. Only the RI75 analogue showed upregulation of SDF-1, a crucial factor in tissue regeneration. Compared to curcumin, the DMAM and RI75 analogues also upregulated runx2 and vegf, both associated with osteodifferentiation. The RI75 analogue demonstrated greater mineralization than curcumin, and both promoted more nodule formation than the untreated control. Our data suggest that the curcumin analogue RI75 at 50 μM presents similar toxicity but enhanced biological activity compared to natural curcumin, making it a promising substance for material biomodifications.

Aims: Oestrogen exerts beneficial cardiovascular effects by binding to specific receptors on various cells to activate nuclear and non-nuclear actions. Oestrogen receptor α (ERα) non-nuclear signalling confers protection against heart failure remodelling, involving myocardial cyclic guanosine monophosphate (cGMP)-cGMP-dependent protein kinase G (PKG) activation; however, its tissue-specific role remains elusive. Herein, we examine the cell type-specific role of ERα non-nuclear signalling in oestrogen-conferred protection against heart failure., Methods and Results: We first assessed the tissue-specific impacts of ERα on the cardiac benefits derived from oestrogen, utilizing endothelial ERα deletion (ERαf/f/Tie2Cre+) and myocyte ERα deletion (ERαf/f/αMHCCre+) female mice. Female mice were ovariectomized and the effect of estradiol (E2) was assessed in hearts exposed to 3 weeks of pressure overload [transverse aortic constriction (TAC)]. E2 failed to improve cardiac function in ERαf/f/Tie2Cre+ TAC hearts but provided benefits in ERαf/f/αMHCCre+ TAC hearts, indicating that endothelial ERα is essential. We next assessed the role of non-nuclear signalling in endothelial cells (ECs), employing animals with endothelial-specific inactivation of ERα non-nuclear signalling (ERαKI/KI/Tie2Cre+). Female ovariectomized mice were supplemented with E2 and subjected to 3-week TAC. ERαKI/KI/Tie2Cre+TAC hearts revealed exacerbated cardiac dysfunction and reduced myocardial PKG activity as compared to littermate TAC hearts, which were associated with attenuated myocardial induction of vascular endothelial growth factor (VEGF) and angiogenesis as assessed by CD31-stained capillary density. This phenotype of ERαKI/KI/Tie2Cre+was rescued by myocardial PKG activation from chronic treatment with a soluble guanylate cyclase (sGC) stimulator. We performed co-culture experiments to determine endothelial-cardiomyocyte interactions. VEGF induction by E2 in cardiac myocytes required a co-existence of intact endothelial ERα signalling in a nitric oxide synthase-dependent manner. On the other hand, VEGF was induced in myocytes directly with an sGC stimulator in the absence of ECs., Conclusion: An endothelial oestrogen-myocardial cGMP axis stimulates angiogenic response and improves cardiac performance during pressure overload., Competing Interests: Conflict of interest: none declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of the European Society of Cardiology.)

This narrative review explores the relationship between genetics and elite endurance athletes, summarizes the current literature, highlights some novel findings, and provides a physiological basis for understanding the mechanistic effects of genetics in sport. Key genetic markers include ACTN3 R577X (muscle fiber composition), ACE I/D (cardiovascular efficiency), and polymorphisms in PPARA , VEGFA , and ADRB2 , influencing energy metabolism, angiogenesis, and cardiovascular function. This review underscores the benefits of a multi-omics approach to better understand the complex interactions between genetic polymorphisms and physiological traits. It also addresses long-standing issues such as small sample sizes in studies and the heterogeneity in heritability estimates influenced by factors like sex. Understanding the mechanistic relationship between genetics and endurance performance can lead to personalized training strategies, injury prevention, and improved health outcomes. Future studies should focus on standardized classification of sports, replication studies involving diverse populations, and establishing solid physiological associations between polymorphisms and endurance traits to advance the field of sports genetics.

Background: Aneurysms are one of the most common and yet devastating cerebrovascular diseases after rupture. Despite several decades of scientific advancements including the expansion of the endovascular capabilities and noninvasive imaging modalities, no medical treatment exists to date. This failure is likely largely attributed to the complex and multifactorial nature of aneurysm pathophysiology. Recent research has increasingly implicated vascular endothelial growth factor (VEGF) in the development and rupture of intracranial aneurysms. Regarded as one of the most potent inducers of angiogenesis, it is a key factor in vascular wall maintenance, inflammation, and regulation of vascular permeability. Whether abnormal VEGF expression is directly related to aneurysm development or acting merely as an acute phase reactant remains uncertain. No review of the current-state-of-evidence on this topic exists yet., Methods and Results: A systematic literature search was performed following PRISMA guidelines May 2024 that queried PubMed (1946-2024), Wiley Cochrane Library: Central Register of Controlled Trials (1898-2024), Thompson Reuters Web of Science: Citation Index (1900-2024), and Google Scholar (1946-2024). Inclusion criteria encompassed human and animal studies that investigated the relationship of intracranial aneurysm and VEGF. Several human and animal models revealed significantly elevated expression of VEGF in intracranial aneurysm tissue, along with greater levels in the cerebrospinal fluid and systemically. Overexpression has been shown to inhibit endothelial cell migration, proliferation, and induce cell apoptosis. Recently, genetic polymorphisms of VEGF have also been shown to significantly correlate with the presence of intracranial aneurysms, establishing the first genetic link between the two., Conclusions: Despite lacking definitive evidence of a causal relationship, the wealth of supporting data substantiates VEGF as a promising topic for future investigation into aneurysm pathophysiology and as a potential therapeutic target.

The development of different phenotypes of coronary artery (CA) lesions is regulated via many various factors, such as pro-inflammatory agents, zinc-dependent endopeptidases, growth factors and circulating microRNAs (miRs). To evaluate the expression levels of miR-34a, miR-145 and miR-222, tumor necrosis factor α (TNF-α), matrix metalloproteinases (MMP-1, -9, -13 and -14) and vascular endothelial growth factor (VEGF) in patients with different phenotypes of coronary artery disease (CAD): ischemia/angina with non-obstructive coronary arteries (INOCA/ANOCA) and obstructive CAD (oCAD) compared with a control group. This cross-sectional observational study included 157 subjects with a verified CAD diagnosis (51 patients with INOCA, 76 patients with oCAD and 30 healthy volunteers). The expression of miR-34a, miR-145 and miR-222 (RT-PCR) and the levels of VEGF, TNF-α, MMP-1, MMP-9, MMP-13 and MMP-14 (ELISA) were estimated in plasma samples. A higher concentration of MMP-9 was found in oCAD-group samples compared to the INOCA/ANOCA group. The INOCA/ANOCA group was characterized by higher levels of TNF-α. Based on multivariate regression analysis, a mathematical model predicting the type of CA lesion was constructed. MiR-145 was the independent predictor of INOCA/ANOCA ( p = 0.006). Changes in concentrations of MMP-9 and MMP-14 were found in both investigated CAD groups, with MMP-9 levels being significantly higher in obstructive CAD samples than in INOCA/ANOCA, which confirms the role of inflammation in the development of atherosclerosis. A multivariate regression analysis allowed us to achieve a model that can predict the phenotype of stable CAD, and MiR-145 can be assumed as an independent predictor of INOCA/ANOCA.

This genetic association study including 120 patients with type 2 diabetes mellitus (T2DM) and 166 non-diabetic individuals aimed to investigate the association of polymorphisms in the genes GSTM1 and GSTT1 (gene deletion), GSTP1 (rs1695), ACE (rs4646994), ACE2 (rs2285666), VEGF-A (rs28357093), and MTHFR (rs1801133) with the development of T2DM in the population of Goiás, Brazil. Additionally, the combined effects of these polymorphisms and the possible differences between sexes in susceptibility to the disease were evaluated. Finally, machine learning models were integrated to select the main risk characteristics for the T2DM diagnosis. Risk associations were found for the GSTT1-null genotype in the non-stratified sample and females, and for mutant C allele of the VEGF-A rs28357093 polymorphism in the non-stratified sample. Furthermore, an association of heterozygous (AG) and mutant (GG) GSTP1 genotypes was observed when combined with GSTT1-null. Machine learning approaches corroborated the results found. Therefore, these results suggested that GSTT1 and GSTP1 polymorphisms may contribute to T2DM susceptibility in a Brazilian sample.

Background: Hypoxia negatively affects the behavior, growth, reproduction and survival of fish, causing serious economic losses to aquaculture. Rainbow trout (Oncorhynchus mykiss), an important economic fish worldwide, belongs to a hypoxia-sensitive fish species, however, little is known about the regulatory mechanism of microRNAs (miRNAs) under hypoxia stress., Results: Rainbow trout were subjected to hypoxia stress for 3 h (H3h&#95;L), 12 h (H12h&#95;L), 24 h (H24h&#95;L) and 3 h reoxygenation (R3h&#95;L) to systemically evaluate the changes of miRNA expression profiles in liver, and functions of sha-miR-92a&#95;L + 2R + 4 were investigated. We found 17, 144, 57 and 55 differentially expressed (DE) miRNAs in the H3h&#95;L vs. control (N&#95;L), H12h&#95;L vs. N&#95;L, H24h&#95;L vs. N&#95;L and R3h&#95;L vs. N&#95;L comparisons, respectively. Enrichment analysis revealed that the targets of DE miRNAs were significantly enriched in HIF signaling pathway, VEGF signaling pathway, FoxO signaling pathway and glycolysis/gluconeogenesis. Through miRNA-mRNA interaction and weighted gene co-expression network analysis (WGCNA), five key DE miRNAs (sha-miR-92a&#95;L + 2R + 4, ssa-miR-128-3p, ssa-miR-101b-3p&#95;R + 1, ola-miR-199a-5p&#95;R + 2 and tni-miR-199&#95;1ss18CG) were identified, which can target at least two hypoxia-responsive genes, such as vegfaa, ho, glut1a and junb. Functional analysis found that sha-miR-92a&#95;L + 2R + 4 directly regulated vegfaa expression by targeting its 3'-UTR, overexpression of sha-miR-92a&#95;L + 2R + 4 significantly decreased vegfaa expression in rainbow trout liver cells, while opposite results were obtained after transfection of sha-miR-92a&#95;L + 2R + 4 inhibitor. Furthermore, overexpressed sha-miR-92a&#95;L + 2R + 4 promoted rainbow trout liver cell proliferation and inhibited apoptosis., Conclusion: This study deepens our understanding of the crucial roles of miRNAs under hypoxia stress in rainbow trout. These results can contribute to devise strategies for improving rainbow trout survival rate and aquaculture production during hypoxia stress and help speeding up the selective breeding of hypoxia-tolerant rainbow trout., Competing Interests: Declarations. Ethics approval and consent to participate: The animal study was reviewed and approved by the Faculty Animal Policy and Welfare Committee of Gansu Agricultural University (Lanzhou, China; Ethic approval file No. GSAU-Eth-AST-2021-004). All experimental procedures and sample collection methods were performed in accordance with approved guidelines and regulations to ensure animal welfare. Meanwhile, the study is in accordance with ARRIVE guidelines. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

To develop in vitro respiratory models, it is crucial to identify the factors involved in epithelial cell differentiation. In this study, we comprehensively analyzed the effects of air-liquid interface (ALI) culture on epithelial cell differentiation using single-cell RNA sequencing (scRNA-seq). ALI culture induced a pronounced shift in cell composition, marked by a fivefold increase in ciliated cells and a reduction of more than half in basal cells. Transcriptional signatures associated with epithelial cell differentiation, analyzed using iPathwayGuide software, revealed the downregulation of VEGFA and upregulation of CDKN1A as key signals for epithelial differentiation. Our findings highlight the efficacy of the ALI culture for replicating the human lung airway epithelium and provide valuable insights into the crucial factors that influence human ciliated cell differentiation.

Purpose: Intravitreal delivery of therapeutic transgenes to the retina via engineered viral vectors can provide sustained local concentrations of therapeutic proteins and thus potentially reduce the treatment burden and improve long-term vision outcomes for patients with neovascular (wet) age-related macular degeneration (AMD), diabetic macular edema (DME), and diabetic retinopathy., Methods: We performed directed evolution in nonhuman primates (NHP) to invent an adeno-associated viral (AAV) variant (R100) with the capacity to cross vitreoretinal barriers and transduce all regions and layers of the retina following intravitreal injection. We then engineered 4D-150, an R100-based genetic medicine carrying 2 therapeutic transgenes: a codon-optimized sequence encoding aflibercept, a recombinant protein that inhibits VEGF-A, VEGF-B, and PlGF, and a microRNA sequence that inhibits expression of VEGF-C. Transduction, transgene expression, and biological activity were characterized in human retinal cells in vitro and in NHPs., Results: R100 demonstrated superior retinal cell transduction in vitro and in vivo compared to AAV2, a commonly used wild-type AAV serotype in retinal gene therapies. Transduction of human retinal pigment epithelial cells in vitro by 4D-150 resulted in dose-dependent transgene expression and corresponding reductions in VEGF-A and VEGF-C. Intravitreal administration of 4D-150 to NHPs was well tolerated and led to robust retinal expression of both transgenes. In a primate model of laser-induced choroidal neovascularization, 4D-150 completely prevented clinically relevant angiogenic lesions at all tested doses., Conclusions: These findings support further development of 4D-150. Clinical trials are underway to establish the safety and efficacy of 4D-150 in individuals with wet AMD and DME.

Ischemic stroke is a sudden onset of neurological deficit resulting from a blockage in cerebral blood vessels, which can lead to brain tissue damage, chronic disability, and increased risk of mortality. Secretome from hypoxic mesenchymal stem cells (SH-MSC) is a potential therapy to improve neurological deficit by increasing the expression of vascular endothelial growth factor (VEGF) and reducing glial fibrillary acidic protein (GFAP). These effects can reduce the infarction area of ischemic stroke. Therefore, the aim of this study was to analyze the effect of 150 μL and 300 μL SH-MSC injection on VEGF and GFAP expression as well as the improvement of infarction area in ischemic stroke animal model. A post-test-only experimental design with consecutive sampling was used, with Rattus norvegicus as subjects. Stromal mesenchymal stem cells (S-MSCs) were isolated from the umbilical cords of rats at 21 days of gestation. Secretome production by the S- MSCs was induced under a hypoxic condition, and subsequently isolated. The resultant secretome was administered to rats subjected to middle cerebral artery occlusion (MCAO) at doses of 150 μL (P1 group) and 300 μL (P2 group). The results showed that the infarction area was reduced in P1 ( p <0.001) and P2 groups ( p <0.001). SH-MSC at a dose of 300 μL increased the expression of VEGF ( p =0.028) and reduced the expression of GFAP ( p =0.001). In conclusion, secretome from hypoxic S-MSC could potentially improve ischemic stroke by upregulating VEGF expression and downregulating GFAP expression., Competing Interests: All the authors declare that there are no conflicts of interest., (© 2024 The Author(s).)

Chronic limb-threatening ischemia (CLTI) is the most severe manifestation of peripheral arterial disease (PAD) and imposes a significantly high burden due to its high risk of mortality and amputation. Revascularization is the first-line treatment for CLTI; however, the amputation rate remains high, and approximately one-third of patients are not eligible for this treatment. Therefore, there is an urgent need for more effective therapeutic strategies. The aim of this study was to investigate the effects and mechanisms of human umbilical vein endothelial cells (HUVECs)-derived exosomes on neovascularization and the degree of necrosis in a hindlimb ischemia model and to study the biological processes underlying their mechanisms. This is an in vivo experimental study with a post-test-only control group design. Forty BALB/c mice were randomized to receive injections of exosomes, conditioned media, and phosphate-buffered saline (PBS) one day after unilateral double ligation. A sham-operated group was also included as a control. Capillary density, arteriole lumen diameter, and histopathological necrosis were measured after seven days, while clinical necrosis was observed daily. MicroRNA profiling, in silico analysis, and transcriptomic analysis of vascular endothelial growth factor (VEGF) mRNA expression were performed to determine the possible biological processes. No amputation was found in the exosome group, as well as in the conditioned media and sham-operated groups, compared to three out of seven mice (43%) in the PBS group. The capillary density was higher in the exosome than in the PBS group ( p  = 0.026). The arteriole lumen diameter in the exosome group was larger than in the PBS ( p  = 0.033) and sham-operated ( p  = 0.034) groups. The scores of clinical necrosis and histopathological necrosis in the exosome group were lower than the PBS group ( p  = 0.005), while the histopathological necrosis scores were also lower but statistically insignificant. In silico analysis showed improvement in neovascularization and necrosis, possibly through energy regulation, PI3 K/AKT and TGF-β activation, the ubiquitin-proteasome system, and tyrosine kinases receptors. HUVEC exosomes were associated with lower VEGF mRNA expression, which may indicate a more effective compensatory mechanism under ischemic conditions. The exosome group had the lowest VEGF mRNA expression compared to other groups, although the difference was not statistically significant. This study highlights that HUVECs-derived exosomes improve neovascularization and decrease necrosis in a hindlimb ischemia mice model, potentially by modulating several possible mechanisms., Competing Interests: The authors declare no potential conflict of interest with respect to the research, authorship, and/or publication of this article., (© 2024 The Author(s).)

Objective To investigate the effect of serum containing Xinfeng capsule (XFC) on the angiogenesis of human umbilical vein endothelial cells (HUVEC) induced by rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) and its mechanism of action. Methods An in vitro co-culture model of RA-FLS and HUVEC was established. Serum containing XFC was prepared by oral gavage of SD rats. CCK-8 was used to screen the optimal co-culture ratio and XFC serum concentration. The lncRNA HOTAIR overexpression plasmid (pcDNA3.1-lncRNA HOTAIR), along with the negative control group, were constructed and transfected into RA-FLS. The experiments were done in HUVEC control group, model group (co-culture of HUVEC and RA-FLS), XFC group (co-culture of RA-FLS treated with 200 mL/L XFC), HOTAIR negative control group (co-culture of RA-FLS transfected with pcDNA3.1-NC), HOTAIR overexpression group (co-culture of RA-FLS transfected with pcDNA3.1-lncRNA HOTAIR), and XFC-treated HOTAIR overexpression group (co-culture of RA-FLS transfected with pcDNA3.1-lncRNA HOTAIR and treated with 200 mL/L XFC). The proliferation ability of HUVEC was detected by CCK-8 method. The migration ability of HUVEC was detected by Transwell TM method. The tube formation ability of HUVEC was detected by tubule formation assay. The expression of CD34 and CD105 in HUVEC was detected by flow cytometry. The expressions of lncRNA HOTAIR, miR-126-3p, phosphatidylinositol 3-kinase (PI3K), PI3K receptor 2 (PIK3R2), AKT, vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) mRNA in HUVEC were detected by real-time quantitative PCR. The protein expressions of PI3K, AKT, p-AKT, VEGF, and bFGF in HUVEC were detected by Western blot and immunofluorescence technique. Results The results of CCK-8 method showed that the optimal treatment ratio and time of RA-FLS and HUVEC co-culture were 5:1 and 48 h respectively. The optimal intervention concentration and time of XFC were 200 mL/L and 48 h. Compared with the control group, the proliferation, migration, tube-forming ability and CD34 and CD105 levels of HUVEC in the model group were significantly improved, the expressions of lncRNA HOTAIR, PIK3R2, VEGF, bFGF, PI3K, AKT and p-AKT were significantly upregulated, and miR-126-3p was significantly downregulated. Compared with the model group, the proliferation, migration, tube-forming ability and CD34 and CD105 levels of HUVEC in the XFC group were significantly decreased, the expressions of lncRNA HOTAIR, PIK3R2, VEGF, bFGF, PI3K, AKT and p-AKT were significantly downregulated, while the expression of miR-126-3p was significantly upregulated. Compared with the HOTAIR negative control group, in the HOTAIR overexpression group, the proliferation, migration, tube-forming ability and CD34 and CD105 levels of HUVECs were significantly increased, the expressions of lncRNA HOTAIR, PIK3R2, VEGF, bFGF, PI3K, AKT and p-AKT were significantly upregulated, and the expression of miR-126-3p was significantly downregulated. Compared with the HOTAIR overexpression group, the proliferation, migration, tube-forming ability and CD34 and CD105 levels of HUVECs in the HOTAIR overexpression group treated with XFC were significantly downregulated, the expressions of lncRNA HOTAIR, PIK3R2, VEGF, bFGF, PI3K, AKT and p-AKT were significantly downregulated, and the expression of miR-126-3p was significantly upregulated. Conclusion XFC-containing serum may play a therapeutic role by inhibiting the expression of lncRNA HOTAIR/PI3K/AKT pathway, reducing the expression levels of VEGF and bFGF, and alleviating synovial angiogenesis induced by RA-FLS to exert therapeutic effect.