Novel strategies in HPV­16­related cervical cancer treatment: An in vitro study of combined siRNA-E5 with oxaliplatin and ifosfamide chemotherapy.

BACKGROUND: Cervical cancer, primarily caused by HPV infection, remains a global health concern. Current treatments face challenges including drug resistance and toxicity. This study investigates combining E5-siRNA with chemotherapy drugs, Oxaliplatin and Ifosfamide, to enhance treatment efficacy in HPV-16 positive cervical cancer cells, targeting E5 oncoprotein to overcome limitations of existing therapies. METHODS: The CaSki cervical cancer cell line was transfected with E5-siRNA, and subsequently treated with Oxaliplatin/Ifosfamide. Quantitative real-time PCR was employed to assess the expression of related genes including p53, MMP2, Nanog, and Caspases. Cell apoptosis, cell cycle progression, and cell viability were evaluated using Annexin V/PI staining, DAPI staining, and MTT test, respectively. Furthermore, stemness ability was determined through a colony formation assay, and cell motility was assessed by wound healing assay. RESULTS: E5-siRNA transfection significantly reduced E5 mRNA expression in CaSki cells compared to the control group. The MTT assay revealed that monotherapy with E5-siRNA, Oxaliplatin, or Ifosfamide had moderate effects on cell viability. However, combination therapy showed synergistic effects, reducing the IC50 of Oxaliplatin from 11.42 × 10-8 M (45.36 µg/ml) to 6.71 × 10-8 M (26.66 µg/ml) and Ifosfamide from 12.52 × 10-5 M (32.7 µg/ml) to 8.206 × 10-5 M (21.43 µg/ml). Flow cytometry analysis demonstrated a significant increase in apoptosis for combination treatments, with apoptosis rates rising from 11.02 % (Oxaliplatin alone) and 16.98 % (Ifosfamide alone) to 24.8 % (Oxaliplatin + E5-siRNA) and 34.9 % (Ifosfamide + E5-siRNA). The sub-G1 cell population increased from 15.7 % (Oxaliplatin alone) and 18 % (Ifosfamide alone) to 21.9 % (Oxaliplatin + E5-siRNA) and 27.1 % (Ifosfamide + E5-siRNA), indicating cell cycle arrest. The colony formation assay revealed a substantial decrease in the number of colonies following combination treatment. qRT-PCR analysis showed decreased expression of stemness-related genes CD44 and Nanog, and migration-related genes MMP2 and CXCL8 in the combination groups. Apoptosis-related genes Casp-3, Casp-9, and pP53 showed increased expression following combination therapy, while BAX expression increased and BCL2 expression decreased relative to the control. CONCLUSION: The study demonstrates that combining E5-siRNA with Oxaliplatin or Ifosfamide enhances the efficacy of chemotherapy in HPV-16 positive cervical cancer cells. This synergistic approach effectively targets multiple aspects of cancer cell behavior, including proliferation, apoptosis, migration, and stemness. The findings suggest that this combination strategy could potentially allow for lower chemotherapy doses, thereby reducing toxicity while maintaining therapeutic efficacy. This research provides valuable insights into targeting HPV E5 as a complementary approach to existing therapies focused on E6 and E7 oncoproteins, opening new avenues for combination therapies in cervical cancer treatment.

Irisin in thyroid diseases.

Irisin, a hormone-like adipo-myokine, has garnered considerable attention in recent years for its potential impact in metabolic diseases. Its physiological effects are similar to those of thyroid hormones, prompting numerous investigations into potential correlations and interactions between irisin and thyroid function through various in vitro and animal experiments. However, existing studies suggest that the relationship between irisin and thyroid diseases is highly complex and multifaceted. In this paper, we have summarized the research results on serum irisin and thyroid function, providing an overview of advancements and constraints in current research on irisin and thyroid hormones. The aim is to offer insights and directions for future clinical trials in this field.

A novel stoploss mutation CYB5R3 c.906A>G(p.*302Trpext*42) involved in the pathogenesis of hereditary methemoglobinemia.

Recessive congenital methemoglobinemia (RCM) is a hereditary autosomal disorder with an extremely low incidence rate. Here, we report a case of methemoglobinemia type I in a patient with congenital persistent cyanosis. The condition was attributed to a novel compound heterozygous mutation in CYB5R3, characterized by elevated methemoglobin levels (13.4 % of total hemoglobin) and undetectable NADH cytochrome b5 reductase (CYB5R3) activity. Whole-exome sequencing (WES) revealed two heterozygous mutations in CYB5R3: a previously reported pathogenic missense mutation c.611G>A(p.Cys204Tyr) inherited from the father, and a novel stop codon mutation c.906A>G(p.*302Trpext*42) from the mother, the latter mutation assessed as likely pathogenic according to ACMG guidelines. In cells overexpressing the CYB5R3 c.906A>G mutant construct, the CYB5R3 mRNA level was significantly lower than in cells overexpressing the wild-type (WT) CYB5R3 construct. However, there was no significant difference in protein expression levels between the mutant and WT constructs. Notably, an additional protein band of approximately 55 kDa was detected in the mutant cells. Immunofluorescence localization showed that, compared to wild-type CYB5R3, the subcellular localization of the CYB5R3 p.*302Trpext*42 mutant protein did not show significant changes and remained distributed in the endoplasmic reticulum and mitochondria. However, the c.906A>G(p.*302Trpext*42) mutation resulted in increased intracellular reactive oxygen species (ROS) levels and decreased NAD+/NADH ratio, suggesting impaired CYB5R3 function and implicating this novel mutation as likely pathogenic.

MiR-6721-5p as a natural regulator of Meta-VCL is upregulated in the serum of patients with coronary artery disease.

Background: Coronary artery disease (CAD), the leading cause of mortality globally, arises from atherosclerotic blockage of the coronary arteries. Meta-vinculin (meta-VCL), a large spliced isoform of VCL, co-localizes in muscular adhesive structures and plays significant roles in cardiac physiology and pathophysiology. This study aimed to identify microRNAs (miRNAs) regulating meta-VCL expression and investigate the expression alterations of the miRNAs of interest and meta-VCL as potential biomarkers in the serum of CAD patients. Methods: Bioinformatics tools were employed to select miRNAs targeting meta-VCL. Cell-based ectopic expression analysis and a dual-luciferase assay were used to examine the interactions between miRNAs and meta-VCL. An ELISA assessed the concentrations of interleukin-6 (IL-6), IL-10, and tumor necrosis factor-α (TNF-α). MiRNA and meta-VCL expression patterns and biomarker suitability were evaluated in serum samples from CAD and non-CAD individuals using real-time PCR. A cardiac cell-line data set and CAD blood exosome samples were analyzed using bioinformatics and ROC curve analyses, respectively. Results: miR-6721-5p directly interacted with the putative target sites at the 3'-UTR of meta-VCL and regulated its expression. IL-10 and TNF-α concentrations, which may act as anti-inflammatory factors, decreased following miR-6721-5p upregulation and meta-VCL downregulation. Bioinformatics and experimental expression analyses confirmed downregulated meta-VCL expression and upregulated miR-6721-5p expression in CAD samples. ROC curve analysis yielded an AUC score of 0.705 (P = 0.018), indicating the potential suitability of miR-6721-5p as a biomarker for CAD. Conclusions: miR-6721-5p plays a regulatory role in meta-VCL expression and may contribute to CAD development by reducing anti-inflammatory factors. These findings suggest that miR-6721-5p could serve as a novel biomarker in the pathogenesis of CAD.

Structural Maintenance of Chromosomes Complexes.

The Structural Maintenance of Chromosomes (SMC) protein complexes are DNA-binding molecular machines required to shape chromosomes into functional units and to safeguard the genome through cell division. These ring-shaped multi-subunit protein complexes, which are present in all kingdoms of life, achieve this by organizing chromosomes in three-dimensional space. Mechanistically, the SMC complexes hydrolyze ATP to either stably entrap DNA molecules within their lumen, or rapidly reel DNA into large loops, which allow them to link two stretches of DNA in cis or trans. In this chapter, the canonical structure of the SMC complexes is first introduced, followed by a description of the composition and general functions of the main types of eukaryotic and prokaryotic SMC complexes. Thereafter, the current model for how SMC complexes perform in vitro DNA loop extrusion is presented. Lastly, chromosome loop formation by SMC complexes is introduced, and how the DNA loop extrusion mechanism contributes to chromosome looping by SMC complexes in cells is discussed.

Photoelectrochemical sensor for nitrite determination based on the etching of BiOCl/Zn0.5Cd0.5S.

A rapid photoelectrochemical (PEC) sensor was constructed for nitrite detection in food based on the one-step chemical etching strategy of BiOCl/Zn0.5Cd0.5S (BOC/ZCS) nanocomposites by nitrite. BOC/ZCS heterojunction was prepared by a simple coprecipitation method, and it was found that BOC/ZCS showed significant photoelectrochemical (PEC) activity. The results of this study confirmed that the decrease in the photocurrent of the sensor was linked to the etching of ZCS by nitrite under acidic conditions. Under optimized conditions, the BOC/ZCS-based PEC sensor showed good analytical properties for detecting nitrite, with linear ranges of 1-100 µM and 100-600 µM. The detection limit of the sensor was 0.41 µM (S/N = 3). Excellent repeatability, reproducibility, low background noise, and immunity to interference were demonstrated using the proposed system, and satisfactory results were achieved for the nitrite assay using real samples. These results demonstrate a new method for nitrite detection developed using the proposed PEC sensor.

The construction and experimental verification of a 6-LncRNA model based on Lactic acid metabolism in the tumor microenvironment of Wilms tumor.

BACKGROUND: Lactic acid (LA) can promote the malignant progression of tumors through the crosstalk with the tumor microenvironment (TME). However, the function of long non-coding RNAs (lncRNAs) related to LA metabolism in Wilms tumor (WT) remains unclear. METHODS: Gene expression data and clinical data of WT patients were collected from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). Through the ESTIMATE algorithm and Pearson correlation analysis, lncRNAs related to tumor immunity and LA metabolism were screened. Subsequently, Cox regression analysis and Lasso Cox regression analysis were used to construct a model. Furthermore, candidate genes were identified and a competitive endogenous RNA (ceRNA) network was conducted to explore the specific mechanism of characteristic genes. Finally, based on the strong clinical relevance of UNC5B-AS1, its expression and function were experimentally verified. RESULTS: The immune score and stromal score were found to be closely related to the prognosis of WT. Eventually, a prognostic model (TME-LA-LM) consisting of 6 lncRNAs was successfully identified. The model demonstrated favorable predictive ability and accuracy, with significant variation in immune infiltration and drug susceptibility observed between risk groups. Additionally, the study revealed the involvement of 2 candidate genes and 5 microRNAs (miRNAs) in the tumor's development. Notably, UNC5B-AS1 was highly expressed and found to promote the proliferation and migration of tumor cells. CONCLUSION: This study, for the first time, elucidated the prognostic signatures of WT using lncRNAs related to TME and LA metabolism. The fundings of this research offer valuable insights for future studies on immunotherapy, personalized chemotherapy and mechanism research.

Comparative evaluation of single and multiple exposure to PM2.5 in respirable air on cardiac physiology, structure and function in a Wistar rat model.

Many studies have shown the negative relationship between long term exposure to PM2.5 and cardiac dysfunction. Recently, studies have shown that even a single exposure of PM2.5 from air sample in permissible range can induce very mild cardiac pathological changes. In the present study, we revisited the toxic effect of PM2.5 on rat heart by adopting single and multiple exposure durations. Female Wistar rats were exposed to PM2.5 at a concentration of 250 µg/m3 daily for 3 hr for single (1 day) and multiple (7, 14, 21 days) durations. The major pathological changes noted in 21 days exposed myocardium comprised of an elevated ST segment (the segment between the S wave and the T wave), development of cardiac fibrosis, hypertrophy, cardiac injury, tissue inflammation and declined cardiac function. With 14 days exposed heart, the electrocardiograms (ECG),data showed insignificantly declined heart rate and an increased QT (the time from the start of the Q wave to the end of the T wave) interval along with mild fibrosis, hypertrophy and lesser number of TUNEL positive cells. On the other hand, single- and 7-days exposure to PM2.5 did not impart any significant changes in the myocardium. To determine the reversibility potential of PM2.5 induced cardiotoxicity, a washout period of 24 hours was adopted and all observed changes in the myocardium were reversed till day 7, but not in 14- and 21-days exposed samples. Based on the above findings we concluded that PM2.5 associated cardiac dysfunction is the cumulative outcome of ineffective cardiac adaptive and repair process that accumulate additively over the time due to prolonged exposure durations.

A methodological framework for estimating ambient PM2.5 particulate matter concentrations in the UK.

Scientific evidence sustains PM2.5 particles' inhalation may generate harmful impacts on human beings' health; therefore, their monitoring in ambient air is of paramount relevance in terms of public health. Due to the limited number of fixed stations within the air quality monitoring networks, development of methodological frameworks to model ambient air PM2.5 particles is primordial to providing additional information on PM2.5 exposure and its trends. In this sense, this work aims to offer a global easily-applicable tool to estimate ambient air PM2.5 as a function of meteorological conditions using a multivariate analysis. Daily PM2.5 data measured by 84 fixed monitoring stations and meteorological data from ERA5 (ECMWF Reanalysis v5) reanalysis daily based data between 2000 and 2021 across the United Kingdom were attended to develop the suggested approach. Data from January 2017 to December 2020 were employed to build a mathematical expression that related the dependent variable (PM2.5) to predictor ones (sea-level pressure, planetary boundary layer height, temperature, precipitation, wind direction and speed), while 2021 data tested the model. Evaluation indicators evidenced a good performance of model (maximum values of RMSE, MAE and MAPE: 1.80 µg/m3, 3.24 µg/m3, and 20.63%, respectively), compiling the current legislation's requirements for modelling ambient air PM2.5 concentrations. A retrospective analysis of meteorological features allowed estimating ambient air PM2.5 concentrations from 2000 to 2021. The highest PM2.5 concentrations relapsed in the Mid- and Southlands, while Northlands sustained the lowest concentrations.

Adult refugees' perspectives on the impact of trauma and post-migration hardships on learning.

Background: Research shows that adult refugees' well-being and future in the reception country heavily depend on successfully learning the host language. However, we know little about how adult learners from refugee backgrounds experience the impact of trauma and adversity on their learning.Objective: The current study aims to investigate the perspectives of adult refugee learners on whether and how trauma and other adversity affect their learning.Methods: We conducted in-depth interviews with 22 adult refugees (10 women) attending the Norwegian Introduction Programme (NIP). The participants came from six Middle Eastern, Central Asian, and African countries. Two questionnaires were included, one about past stressful life events (SLESQ-Revised), and one about mental health symptoms and current psychological distress following potentially traumatic experiences (PCL-5).Results: Participants held varying beliefs about trauma's impact on learning: that it had a constant impact, that it was situational, or that it had no impact. Other aspects they brought up as having an essential effect on learning and school attendance include psychological burdens from past and present school experiences, and post-migration hardships such as loneliness, depression, ongoing violence, and negative social control. Post-migration trauma and hardships exacerbated the burden of previous trauma and were frequently associated with a greater negative influence on learning.Conclusion: This study adds new insights from adult refugee learners themselves into how post-migration hardships as well as trauma can impact their learning, and the importance of recognising their struggles. A safe space is required for refugees to open up about their difficulties in life and with learning. This knowledge can be used to enhance teaching practices, foster better teacher-student relationships, and inform policy-making decisions, ultimately benefiting both individuals and society.

Adult refugee learners' own perspectives on the impact of trauma on learning varied from constant to situational to no impact at all.Other factors identified as impacting learning and school attendance included, amongst others, psychological burdens from past and present school experiences, ongoing violence, forced family separation, and negative social control.Post-migration trauma and hardships were frequently associated with a greater negative influence on learning than the burden of previous trauma.

Homocysteine decreases VEGF, EGF, and TrkB levels and increases CCL5/RANTES in the hippocampus: Neuroprotective effects of rivastigmine and ibuprofen.

Homocysteine (Hcy) is produced through methionine transmethylation. Elevated Hcy levels are termed Hyperhomocysteinemia (HHcy) and represent a risk factor for neurodegenerative conditions such as Alzheimer's disease. This study aimed to explore the impact of mild HHcy and the neuroprotective effects of ibuprofen and rivastigmine via immunohistochemical analysis of glial markers (Iba-1 and GFAP). Additionally, we assessed levels of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), chemokine ligand 5 (CCL5/RANTES), CX3C chemokine ligand 1 (CX3CL1), and the NGF/p75NTR/tropomyosin kinase B (TrkB) pathway in the hippocampus of adult rats. Mild chronic HHcy was induced chemically in Wistar rats by subcutaneous administration of Hcy (4 mg/kg body weight) twice daily for 30 days. Rivastigmine (0.5 mg/kg) and ibuprofen (40 mg/kg) were administered intraperitoneally once daily. Results revealed elevated levels of CCL5/RANTES and reduced levels of VEGF, EGF, and TrkB in the hippocampus of HHcy-exposed rats. Rivastigmine mitigated the neurotoxic effects of HHcy by increasing TrkB and VEGF levels. Conversely, ibuprofen attenuated CCL5/RANTES levels against the neurotoxicity of HHcy, significantly reducing this chemokine's levels. HHcy-induced neurochemical impairment in the hippocampus may jeopardize neurogenesis, synapse formation, axonal transport, and inflammatory balance, leading to neurodegeneration. Treatments with rivastigmine and ibuprofen alleviated some of these detrimental effects. Reversing HHcy-induced damage through these compounds could serve as a potential neuroprotective strategy against brain damage.

Understanding the emergence of highly pathogenic avian influenza A virus H5N1 in pinnipeds: An evolutionary approach.

Highly pathogenic influenza A virus (HPIAV) H5N1 within the genetic clade 2.3.4.4b has emerged in wild birds in different regions of the world, leading to the death of >70 million birds. When these strains spread to pinniped species a remarkable mortality has also been observed. A detailed genetic characterization of HPIAV isolated from pinnipeds is essential to understand the potential spread of these viruses to other mammalian species, including humans. To gain insight into these matters a detailed phylogenetic analysis of HPIAV H5N1 2.3.4.4b strains isolated from pinniped species was performed. The results of these studies revealed multiple transmission events from birds to pinnipeds in all world regions. Different evolutionary histories of different genes of HPIAV H5N1 2.3.4.4b strains gave rise to the viruses infecting pinnipeds in different regions of the world. European strains isolated from pinnipeds represent a completely different genetic lineage from strains isolated from South American ones. All strains isolated from pinnipeds bear characteristics of a highly pathogenic form for of avian influenza in poultry. Amino acid substitutions, previously shown to confer an adaptive advantage for infecting mammals, were observed in different genes in all pinniped species studied.

Identification of immune-related hub genes in spinal cord injury.

OBJECTIVES: Immune regulation is a pivotal factor in the pathogenesis and repair of spinal cord injury (SCI). This study aims to explore potential immune center genes associated with spinal cord injury. METHODS: The public data set GSE151371 was obtained from the GEO database. The R software package "limma" was used to identify differentially expressed genes (DEGs) in SCI. GO, KEGG and GSEA pathway analyses were performed using the DEGs. The key module genes related to spinal cord injury were selected through WGCNA analysis. Overlapping genes were extracted from WGCNA, DEGs, and immune-related genes. LASSO analysis was employed to identify central genes associated with SCI immunity. Pearson correlation analysis assessed the correlation between hub genes and immune cells in SCI. In addition, we further investigated the hub genes' expression, diagnostic potential, function, and targeted drugs. RESULTS: We have identified three immunity-related hub genes (ABHD5, EDNRB, EDN3). Immune infiltration analysis showed that the hub gene was significantly associated with resting NK cells, M2 macrophages, and monocytes in the immune microenvironment of SCI. ROC analysis demonstrated that these hub genes have favorable diagnostic performance for SCI. Functional analysis revealed that ABHD5 is primarily associated with lipid metabolism pathways, while EDN3 and EDNRB are mainly involved in endothelin, downstream GPCR signaling, and ERK signaling transduction. In addition, we identified six potential targeted drugs based on our findings. CONCLUSIONS: ABHD5, EDNRB, and EDN3 are involved in processes such as SCI progression or repair through immunomodulation and deserve further study.

From silos to synergy: a consortium approach to air pollution and public health in Abu Dhabi.

Financial resources alone cannot guarantee effective public health policy. In Abu Dhabi, massive economic growth in the desert climate resulted in concentrated urbanization and led to challenges in the regulation of air pollution. The Environment Agency in Abu Dhabi commissioned us to scope the regulatory challenges for air pollution. Part of this project relied on the participation and involvement of key stakeholders. We found three barriers: (1) limited appreciation of uncertainties in risk estimates and discussion on the importance of considering control costs and the societal trade-offs between health and wealth inherent in such decisions, (2) compartmentalization of efforts, and (3) challenges to decide how to prioritize risks in policy agendas. We propose a consortium-like approach that brings stakeholders together and places risk, uncertainty, and tradeoffs between health and wealth at the forefront of decision-making. Expected outcomes include improved collaboration and information sharing, strategic prioritization of emission controls, and a better understanding and consideration of uncertainty to guide future public health research.

Placebo-controlled efficacy of 5-HT3 antagonists for postoperative nausea and vomiting prophylaxis in supratentorial craniotomies: A systematic review and comparative meta-analysis of randomized clinical trials.

BACKGROUND: Postoperative nausea and vomiting (PONV) are common and distressing complications following neurosurgical procedures, affecting up to 73 % of patients undergoing craniotomy. Therefore, we aimed to assess the placebo-controlled efficacy of 5-HT3 antagonists to prevent PONV following supratentorial craniotomies. METHODS: We searched Medline, Web of Science, and Embase databases following PRISMA guidelines for RCTs comparing the outcomes of prophylactic use of 5-HT3 antagonists with placebo to prevent PONV following supratentorial craniotomy. We pooled odds ratios (OR) with 95 % confidence intervals with a random-effects model. I2 statistics was used to assess heterogeneity. RESULTS: Five RCTs, comprising 347 patients, of which 145 received a placebo, were included. The analysis identified a lower likelihood of early postoperative vomiting in 5-HT3 antagonists group (OR=0.47; 95 % CI: 0.24-0.91, p<0.05; I2=7 %), a lower likelihood of vomit within the 24-h period in 5-HT3 antagonists group (OR=0.27; 95 % CI: 0.15-0.48, p<0.01; I2=40 %), a lower likelihood of nausea within the 24-h period in 5-HT3 antagonists group (OR=0.47; 95 % CI: 0.28-0.72, p<0.01; I2=34 %), and a lower likelihood of rescue interventions in 5-HT3 antagonists group (OR = 0.18; 95 % CI: 0.10-0.34; I2 = 0 %. Subgroup analyses focusing on ondansetron also identified a lower likelihood of nausea and vomiting within the 24-h period in the 5-HT3 antagonist group. CONCLUSION: This systematic review and meta-analysis identified that 5-HT3 antagonists are effective in preventing PONV in the postoperative period following supratentorial craniotomy when compared to placebo. Our findings provide synthesized and robust evidence derived from randomized studies to support the use of 5-HT3 antagonists in clinical practice.

RFC2 may contribute to the pathogenicity of Williams syndrome revealed in a zebrafish model.

Williams syndrome (WS) is a rare multisystemic disorder caused by recurrent microdeletions on 7q11.23, characterized by intellectual disability, distinctive craniofacial and dental features, and cardiovascular problems. Previous studies have explored the roles of individual genes within these microdeletions in contributing to WS phenotypes. Here, we report five patients with WS with 1.4 Mb-1.5 Mb microdeletions that include RFC2, as well as one patient with a 167 kb microdeletion involving RFC2 and six patients with intragenic variants within RFC2. To investigate the potential involvement of RFC2 in WS pathogenicity, we generate a rfc2 knockout (KO) zebrafish using CRISPR-Cas9 technology. Additionally, we generate a KO zebrafish of its paralog gene, rfc5, to better understand the functions of these RFC genes in development and disease. Both rfc2 and rfc5 KO zebrafish exhibit similar phenotypes reminiscent of WS, including small head and brain, jaw and dental defects, and vascular problems. RNA-seq analysis reveals that genes associated with neural cell survival and differentiation are specifically affected in rfc2 KO zebrafish. In addition, heterozygous rfc2 KO adult zebrafish demonstrate an anxiety-like behavior with increased social cohesion. These results suggest that RFC2 may contribute to the pathogenicity of Williams syndrome, as evidenced by the zebrafish model.

YTHDF1-regulated ALOX5 in retinal pigment epithelial cells under hypoxia enhances VEGF expression and promotes viability, migration, and angiogenesis of vascular endothelial cells.

Upregulation of vascular endothelial growth factor (VEGF) and enhanced angiogenesis have been implicated in the severe progression of age-related macular degeneration (AMD). Abnormal arachidonate 5-lipoxygenase (ALOX5) is associated with AMD pathogenesis. However, no reports have shown the causal role of ALOX5 in angiogenesis during AMD. In the present study, ARPE-19 cells were exposed to hypoxia, an inducer of VEGF expression. Potential proteins implicated in AMD progression were predicted using bioinformatics. RNA affinity antisense purification-mass spectrometry (RAP-MS) was applied to identify the binding proteins of ALOX5 3'UTR. Expression of ALOX5 and YTH N6-methyladenosine RNA-binding protein 1 (YTHDF1) was detected by qRT-PCR and western blotting. VEGF expression and secretion were assessed by immunofluorescence and ELISA, respectively. The chicken embryo chorioallantoic membrane (CAM) was used to analyze the effect of ALOX5 on angiogenesis. RNA stability was assayed using the Actinomycin D assay. The results show that hypoxia promoted cell growth and increased VEGF expression in ARPE-19 cells. ALOX5 was associated with AMD progression, and hypoxia upregulated ALOX5 expression in ARPE-19 cells. ALOX5 silencing reduced VEGF expression induced by hypoxia in ARPE-19 cells. Moreover, the conditioned medium of ALOX5-silenced ARPE-19 cells could suppress the viability and migration of HUVECs and diminish angiogenesis in the CAM. Furthermore, YTHDF1 was validated to bind to ALOX5 3'UTR, and YTHDF1 promoted ALOX5 expression by elevating the stability of ALOX5 mRNA. In conclusion, our findings demonstrate that YTHDF1-regulated ALOX5 increases VEGF expression in hypoxia-exposed ARPE-19 cells and enhances the viability, migration, and angiogenesis of vascular endothelial cells.

LncRNA TUG1 regulates mir-34a-5p / SIRT6 to participate in benzene-induced hematotoxicity through PI3K / AKT /mTOR signaling pathway.

LncRNA TUG1 plays pivotal roles in various diseases. However, its exact roles in benzene - induced hematotoxicity remain unclear. Herein, we aimed to investigate the role and mechanism of TUG1 in hematoxic injuries caused by benzene. In the current study, TUG1 was found dramatically decreased in WBCs of benzene exposure workers and negatively correlated with benzene exposure duration and urine SPMA. In vitro assays demonstrated that TUG1 overexpression attenuated 1,4-BQ-caused suppression of cell viability and proliferation, and promotion of ROS generation and apoptosis via PI3K/AKT/mTOR pathway. Bioinformatic prediction and molecular assay validated miR-34a-5p was negatively regulated by TUG1. The miR-34a-5p was upregulated in 1,4-BQ treated cells and downregulated in TUG1 overexpression cells. Moreover, miR-34a-5p upregulation partially reversed the protective effects of TUG1 overexpression on 1,4-BQ - caused cytotoxicity. Furthermore, SIRT6 was a downstream target gene of miR-34a-5p, whose expression was reduced in miR-34a-5p upregulation cells and elevated in TUG1 overexpression cells. Upregulated SIRT6 could counteract accelerated cytotoxicity mediated by miR-34a-5p upregulation after 1,4-BQ treatment. Taken together, our study revealed that the critical role of the TUG1/miR-34a-5p/SIRT6 axis in benzene-caused hematotoxicity, and provided scientific basis for further understanding the epigenetic regulatory mechanisms underlying benzene hematotoxicity.

Circ_0104873 promotes osteoarthritis progression via miR-875-5p/NOTCH3/Notch signaling pathway.

Osteoarthritis (OA) is the most common joint disease with high prevalence and incidence. Increasing reports has indicated that circular RNAs (circRNAs) are implicated in OA progression. Nevertheless, the roles and functions of most circRNAs in OA remain to be elucidated. In this study, we emphatically discussed circ-IQGAP1 (circ_0104873) in OA. Firstly, we discovered that circ_0104873 was dramatically overexpressed during osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Several functional assays demonstrated that circ_0104873 inhibition repressed BMSCs proliferation and osteogenic differentiation. Moreover, mechanism assays also revealed that circ_0104873 sponged microRNA-875-5p (miR-875-5p) to up-regulate notch receptor 3 (NOTCH3), thereby activating the Notch signaling pathway. Rescue assays disclosed that circ_0104873 contributed to the development of OA via targeting miR-875-5p/NOTCH3 axis. In conclusion, circ_0104873 promoted the progression of OA by miR-875-5p/NOTCH3/Notch signaling pathway, which might provide a promising target for OA treatment.