RESUMO
Atherosclerosis treatments by gene regulation are garnering attention, yet delivery of gene cargoes to atherosclerotic plaques remains inefficient. Here, we demonstrate that assembly of therapeutic oligonucleotides into a three-dimensional spherical nucleic acid nanostructure improves their systemic delivery to the plaque and the treatment of atherosclerosis. This noncationic nanoparticle contains a shell of microRNA-146a oligonucleotides, which regulate the NF-κB pathway, for achieving transfection-free cellular entry. Upon an intravenous injection into apolipoprotein E knockout mice fed with a high-cholesterol diet, this nanoparticle naturally targets class A scavenger receptor on plaque macrophages and endothelial cells, contributing to elevated delivery to the plaques (â¼1.2% of the injected dose). Repeated injections of the nanoparticle modulate genes related to immune response and vascular inflammation, leading to reduced and stabilized plaques but without inducing severe toxicity. Our nanoparticle offers a safe and effective treatment of atherosclerosis and reveals the promise of nucleic acid nanotechnology for cardiovascular disease.
Assuntos
Aterosclerose , MicroRNAs , Nanopartículas , Placa Aterosclerótica , Animais , Aterosclerose/tratamento farmacológico , Aterosclerose/genética , Células Endoteliais/metabolismo , Camundongos , Camundongos Knockout , MicroRNAs/genética , MicroRNAs/uso terapêutico , NF-kappa B/genética , NF-kappa B/metabolismo , Nanopartículas/química , Nanopartículas/uso terapêutico , Oligonucleotídeos/uso terapêutico , Placa Aterosclerótica/metabolismo , Receptores Depuradores/metabolismoRESUMO
With the increasing energy demand, oil is still an important fuel source worldwide. The chemical flooding process is used in petroleum engineering to increase the recovery of residual oil. As a promising enhanced oil-recovery technology, polymer flooding still faces some challenges in achieving this goal. The stability of a polymer solution is easily affected by the harsh reservoir conditions of high temperature and high salt, and the influence of the external environment such as high salinity, high valence cations, pH value, temperature and its own structure is highlighted. This article also involves the introduction of commonly used nanoparticles, whose unique properties are used to improve the performance of polymers under harsh conditions. The mechanism of nanoparticle improvement on polymer properties is discussed, that is, how the interaction between them improves the viscosity, shear stability, heat-resistance and salt-tolerant performance of the polymer. Nanoparticle-polymer fluids exhibit properties that they cannot exhibit by themselves. The positive effects of nanoparticle-polymer fluids on reducing interfacial tension and improving the wettability of reservoir rock in tertiary oil recovery are introduced, and the stability of nanoparticle-polymer fluid is described. While analyzing and evaluating the research on nanoparticle-polymer fluid, indicating the obstacles and challenges that still exist at this stage, future research work on nanoparticle-polymer fluid is proposed.
Assuntos
Nanopartículas , Petróleo , Polímeros/química , Fenômenos Químicos , Viscosidade , Nanopartículas/químicaRESUMO
We present a self-therapeutic nanoparticle for topical delivery to epidermal keratinocytes to prevent and treat psoriasis. Devoid of known chemical or biological antipsoriatic drugs, this sub-15 nm nanoparticle contains a 3 nm gold core and a shell of 1000 Da polyethylene glycol strands modified with 30% octadecyl chains. When it is applied to imiquimod-induced psoriasis mice without an excipient, the nanoparticle can cross the stratum corneum and preferentially enter keratinocytes. Applying the nanoparticles concurrently with imiquimod prevents psoriasis and downregulates genes that are enriched in the downstream of the interleukin-17 signaling pathway and linked to epidermis hyperproliferation and inflammation. Applying the nanoparticles after psoriasis is established treats the psoriatic skin as effectively as standard steroid and vitamin D analog-based therapy but without hair loss and skin wrinkling. The nanoparticles do not accumulate in major organs or induce long-term toxicity. Our nanoparticle offers a simple, safe, and effective alternative for treating psoriasis.
Assuntos
Nanopartículas Metálicas , Nanopartículas , Psoríase , Animais , Modelos Animais de Doenças , Ouro , Imiquimode , Queratinócitos , Camundongos , Psoríase/tratamento farmacológicoRESUMO
Acid rain is a global environmental problem that mobilizes heavy metals in soils, while the distribution and geochemical fraction of heavy metals during acid rain infiltration in heterogeneous soils are still unclear. In this study, we performed column experiments to investigate the distribution and geochemical fraction of Cu, Pb, Ni and Cd in heterogeneously layered soils during continuous acid rain infiltration. Chloride ion used as a conservative tracer was found to be uniformly distributed during acid rain infiltration, showing insignificant preferential flow effects in the column. In contrast, however, the distribution of heavy metals was highly non-uniform, especially in the silty soil at the lower part of the column, indicating a heterogeneous distribution of adsorption capacity. In addition, in the control experiments with neutral rain infiltration, uniform distribution of heavy metals was observed, indicating that the heterogeneous distribution of adsorption coefficient during acid rain infiltration was mainly caused by different pH buffering capacities. A numerical model considering water flow and solute transport was developed, where the average water-solid distribution coefficient (Kd) in Layer 2 was only 1.5-12.5% of that in Layer 1 during acid rain infiltration. The model could predict the variation of heavy metal concentrations in soil with the majority of error less than 35%, confirming that different Kd induced the heterogeneous distribution of heavy metals. In addition, the geochemical fraction of heavy metals in the upper coarse sand layer remained stable, while the acid-extractable fractions in the lower loam and silt loam layer gradually increased. Our findings suggest that soil heterogeneity, especially chemical heterogeneity affected by rainfall acidity, has an important influence on the infiltration, migration and geochemical fraction of heavy metals in soils. This study could help guide the risk assessment of heavy metal-contaminated sites that were polluted by acid rain or landfill leachate.
Assuntos
Chuva Ácida , Metais Pesados , Poluentes do Solo , Poluentes Químicos da Água , Cádmio , China , Cloretos , Monitoramento Ambiental , Chumbo , Metais Pesados/análise , Areia , Solo , Poluentes do Solo/análise , ÁguaRESUMO
In diabetes-induced complications, inflammatory-mediated endothelial dysfunction is the core of disease progression. Evidence shows that kakonein, an isoflavone common in Pueraria, can effectively treat diabetes and its complications. Therefore, we explored whether kakonein protects cardiovascular endothelial function by inhibiting inflammatory responses. In this study, C57BL/6J mice were injected with streptozocin to establish a diabetes model and treated with kakonein or metformin for 7 days. The protective effect of kakonein on cardiovascular endothelial junctions and NLRP3 inflammasome activation was verified through immunofluorescence and ELISA assay. In addition, the regulation of autophagy on the NLRP3 inflammasome was investigated through Western blot, immunofluorescence and RT-qPCR. Results showed that kakonein restored the function of endothelial junctions and inhibited the assembly and activation of the NLRP3 inflammasome. Interestingly, kakonein decreased the expression of NLRP3 inflammasome protein by not reducing the transcriptional levels of NLRP3 and caspase-1. Kakonein activated autophagy in an AMPK-dependent manner, which reduced the activation of the NLRP3 inflammasome. In addition, kakonein inhibited both hyperglycaemia-induced cardiovascular endothelial junction dysfunction and NLRP3 inflammasome activation, similar to autophagy agonist. Our findings indicated that kakonein exerts a protective effect on hyperglycaemia-induced chronic vascular disease by regulating the NLRP3 inflammasome through autophagy.
Assuntos
Angiopatias Diabéticas/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Endotélio Vascular/efeitos dos fármacos , Isoflavonas/uso terapêutico , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Vasodilatadores/uso terapêutico , Quinases Proteína-Quinases Ativadas por AMP/metabolismo , Animais , Autofagia , Células Cultivadas , Angiopatias Diabéticas/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Endotélio Vascular/metabolismo , Inflamassomos/metabolismo , Isoflavonas/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteólise , Vasodilatadores/farmacologiaRESUMO
Quorum sensing (QS) plays an important role in the intensive communication between plants and microbes in the rhizosphere during the phytoremediation. This study explored the influence of the root exudates of hyperaccumulator Sedum alfredii on Pseudomonas aeruginosa based on QS. The effects of the components of root exudates, genes expression and transcription regulation of QS system (especially the las system) in Pseudomonas aeruginosa wild-type strain (WT) and rhl system mutant strain (ΔrhlI) were systematically analyzed and discussed. The WT and ΔrhlI exposed to gradient root exudates (0×, 1×, 2×, 5× and 10×) showed a concentration-corrective inhibition on protease production, with the inhibition rates of 51.4-74.5% and 31.2-50.0%, respectively. Among the components of the root exudates of Sedum alfredii, only thymol had an inhibition effects to the root exudates on the activity of protease and elastase. The inhibition rates of 50 µmol/L thymol on protease and elastase in WT were 44.7% and 24.3%, respectively, which was consistent with the variation in ΔrhlI. The gene expression of lasB declined 36.0% under the 1× root exudate treatment and 73.0% under the 50 µmol/L thymol treatment. Meanwhile, there was no significant impact on N-3-oxo-dodecanoyl-L-homoserine lactone signal production and the gene expression of lasI and lasR. Therefore, thymol from Sedum alfredii root exudates could inhibit the formation of protease and elastase in Pseudomonas aeruginosa by suppressing the expression of lasB, without any significant influence on the main las system as a potential natural QS inhibitor.
Assuntos
Exsudatos de Plantas/toxicidade , Pseudomonas aeruginosa/efeitos dos fármacos , Percepção de Quorum/efeitos dos fármacos , Sedum , 4-Butirolactona/análogos & derivados , Proteínas de Bactérias/metabolismo , Exsudatos e Transudatos/metabolismo , Regulação Bacteriana da Expressão Gênica , Pseudomonas aeruginosa/metabolismoRESUMO
Advances in nanotechnology have empowered the design of bionanomaterials by assembling different types of natural biomolecules (e.g., nucleic acids, proteins, and lipids) as building blocks into nanoparticles (NPs) of 1-100 nm in diameter. Such bionanomaterials form the basis of useful nanomedicine applications, such as targeted delivery, gene regulation, molecular diagnostics, and immunomodulation. To achieve optimal performance in these applications, it is imperative that the NPs be delivered effectively to the organs, tissues, and cells of interest. A rational approach to facilitating the delivery of NPs is to develop a detailed and comprehensive understanding in their fundamental interactions with the biological system (or nano-bio interactions). Rigorous nano-bio research can provide mechanistic insights for circumventing the bottlenecks associated with inefficient and nonspecific delivery of NPs, catalyzing the clinical translation of nanomedicines. Cationic liposomes and lipid NPs are conventional carriers of therapeutic cargoes into cells due to their high ability to penetrate the cell membrane, a barrier comprised by an anionic phospholipid bilayer. Yet, cationic NPs tend to cause cytotoxicity and immune responses that may hamper their clinical translation. Contrary to cationic NPs, non-cationic NPs (be they near-neutral or anionic in surface charge) generally exhibit higher biocompatibility but enter mammalian cells in much less pronounced amounts. Intriguingly, some types of non-cationic NPs exhibit high biocompatibility and cellular uptake properties, all attractive features for intracellular delivery. In this Account, we present our studies of the interactions of non-cationic bionanomaterials with cells (or nano-cell interactions). To start with, we introduce the use of near-neutral poly(ethylene glycol)-coated NPs for probing the roles of two rarely explored physicochemical parameters on cellular uptake, namely, extracellular compression and alkylation. We next present the nano-cell interactions of two representative types of anionic bionanomaterials that effectively enter mammalian cells and have found widespread applications in the past decade, including DNA-coated NPs and polydopamine (PDA)-coated NPs. In our cell-based studies, we dissect the route of intracellular trafficking, pathway proteins that dictate cellular uptake, and trafficking of NPs. We further touch on our recent quantitative analysis of the cellular-level distribution of NPs in various organs and tissues of diseased animal models. Our results offer important design rules of NPs for achieving effective intracellular delivery and may even guide us to explore nanomedicine applications that we did not conceive before, such as using DNA-coated NPs for targeting atherosclerotic plaques and PDA-coated plasmonic nanoworms for photothermal killing of cancer cells. We conclude with our perspectives in elucidating nano-bio interactions via a reductionist approach, calling for closer attention to the role of functional groups and more refined studies on the organelle-level distribution of NPs and the genetic basis of in vivo distribution of NPs.
Assuntos
Transporte Biológico/fisiologia , Nanopartículas/metabolismo , Animais , Linhagem Celular Tumoral , DNA/química , Endocitose/fisiologia , Feminino , Ouro/química , Humanos , Indóis/química , Masculino , Camundongos , Polietilenoglicóis/química , Polímeros/químicaRESUMO
Posterior capsule opacification (PCO) is a common long-term complication of modern cataract surgery. The epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs) is a crucial process in the development of PCO. The purpose of this study is to investigate the role of microRNA-34a (miR-34a) in the regulation of EMT and its target gene. Human LECs were treated with TGFß2 to induce EMT as a model for PCO. The mRNA levels of miR-34a and EMT markers were examined by real-time quantitative polymerase chain reaction (qPCR). The expression level of miR-34a was downregulated, whereas that of Notch1 was upregulated in TGFß2-induced EMT of LECs. Overexpression of miR-34a by transfection with miR-34a inhibited EMT of LECs and reduced the expression of Notch1; while, inhibition of miR-34a upregulated the expression of both Notch1 and its ligand Jagged1 in LECs. Luciferase reporter assays revealed that Notch1 gene was direct target of miR-34a. Moreover, DAPT, a specific inhibitor of Notch signaling pathway, reversed LEC-EMT. In addition, the expression level of miR-34a was downregulated, whereas that of Notch1 was upregulated in capsular opacification from cataract samples. MiR-34a can negatively regulate EMT of LECs by targeting Notch1. Therefore, miR-34a/Notch1 could serve as a potential therapeutic approach for the treatment of PCO.
Assuntos
Opacificação da Cápsula/metabolismo , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Cristalino/efeitos dos fármacos , MicroRNAs/fisiologia , Cápsula Posterior do Cristalino/metabolismo , Receptor Notch1/metabolismo , Adulto , Idoso , Western Blotting , Opacificação da Cápsula/patologia , Células Cultivadas , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Cristalino/metabolismo , Masculino , Pessoa de Meia-Idade , Cápsula Posterior do Cristalino/patologia , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Receptor Notch1/genética , Transfecção , Fator de Crescimento Transformador beta2/farmacologiaRESUMO
OBJECTIVE: To detect pathogenic gene variants in two Chinese families with cone-rod dystrophy(CORD). METHODS: After the informed consent and comprehensive ophthalmic examinations for the patients, 3 mL peripheral blood was taken from the patients' blood vessel and DNA was extracted. The DNA was sequenced by whole-exome sequencing technology and variants were analyzed. RESULTS: Two novel compound heterozygous AIPL1 variants were detected in two patients, which were c.923T to C (p.L308P) and c.421C to T (p.Q141X) variants in Family 1, c.572T to C (p.L191P) and c.421C to T (p.Q141X) in Family 2. CONCLUSION: The results supported that AIPL1 gene variants are the main cause of the two CORD families. Whole-exome sequencing technology is a useful tool in the clinical differentiated diagnosis and genetic counseling for CORD patients.
Assuntos
Proteínas de Transporte/genética , Distrofias de Cones e Bastonetes/genética , Proteínas do Olho/genética , Proteínas Adaptadoras de Transdução de Sinal , Povo Asiático , Humanos , Mutação , Linhagem , Sequenciamento do ExomaRESUMO
Purpose: Adenoid cystic carcinoma (ACC) in the lacrimal gland is a rare malignancy. P16 is encoded by the CDKN2A gene, which is recognized as a tumor suppressor due to its inactivation in many types of tumors. However, p16 overexpression is also linked to adverse tumor parameters. These contradictory observations have also been confirmed in ACCs in the salivary glands. Furthermore, evidence of human papilloma virus (HPV) infection is found in a proportion of ACCs in the salivary glands. P16 is often overexpressed in HPV-related squamous cell carcinoma in parallel. To our knowledge, the role of p16 and HPV in ACCs in the lacrimal gland is still unknown. Methods: Twenty-one ACCs in the lacrimal gland and ten matched healthy lacrimal glands were studied. P16 was detected with immunohistochemistry (IHC), and HPV was detected with in situ hybridization (ISH) and PCR in all cases. Other cell cycle proteins were also detected with IHC, including cyclin D1 and Ki67. The methylation status of the p16 promoter was detected with methylation-specific PCR (MSP) to further investigate the regulation of p16 expression. Results: The expression rates of p16 (47.6%, 10/21), cyclin D1 (100%, 21/21), and Ki67 (52.4%, 11/21) were increased in ACCs compared to healthy lacrimal glands (negative). The results showed p16 expression was limited to the inner ductal epithelial cells in the majority of the tubular and cribriform patterns. In solid ACCs, p16 was uniformly positive. HPV was negative in all 21 cases with ISH and PCR. P16 overexpression was associated with cyclin D1 overexpression (p=0.013). Only 13 cases were tested successfully with MSP. The expression rate of p16 methylation was 23.1% (3/13) of the ACCs. Compared with primary ACCs, recurrent ACCs showed higher p16, cyclin D1, and Ki67 expression (p=0.011, p=0.026, p=0.049, respectively). Conclusions: In summary, p16 overexpression was cell-type dependent in ACCs in the lacrimal gland, while HPV infection was negative. P16 overexpression was unrelated to HPV infection. The mechanism of p16 overexpression needs to be further investigated in ACCs in the lacrimal gland.
Assuntos
Carcinoma Adenoide Cístico/genética , Ciclina D1/genética , Inibidor de Quinase Dependente de Ciclina p18/genética , Neoplasias Oculares/genética , Regulação Neoplásica da Expressão Gênica , Antígeno Ki-67/genética , Adolescente , Adulto , Idoso , Carcinoma Adenoide Cístico/metabolismo , Carcinoma Adenoide Cístico/mortalidade , Carcinoma Adenoide Cístico/patologia , Estudos de Casos e Controles , Ciclina D1/metabolismo , Inibidor p16 de Quinase Dependente de Ciclina , Inibidor de Quinase Dependente de Ciclina p18/metabolismo , Neoplasias Oculares/metabolismo , Neoplasias Oculares/mortalidade , Neoplasias Oculares/patologia , Feminino , Humanos , Antígeno Ki-67/metabolismo , Aparelho Lacrimal/metabolismo , Aparelho Lacrimal/patologia , Masculino , Pessoa de Meia-Idade , Papillomaviridae , Infecções por Papillomavirus , Análise de SobrevidaRESUMO
BACKGROUND: Pulmonary arterial smooth muscle cell (PASMC) proliferation in response to hypoxia plays an important role in the vascular remodelling that occurs in hypoxic pulmonary hypertension. MicroRNAs (miRs) are emerging as important regulators in the progression of pulmonary hypertension. In this study, we investigated whether the expression of miR-17-5p is modulated by hypoxia and is involved in the hypoxia-induced proliferation of PASMCs. METHODS: Human PASMCs were cultured under hypoxic conditions. miR-17-5p expression was determined by real-time RT-PCR. A BrdU incorporation assay and time-lapse recording were utilized to determine cell proliferation and migration. RESULTS: PASMC proliferation was increased by moderate hypoxia (3% oxygen) but was reduced by severe hypoxia (0.1% oxygen) after 48 h. Moderate hypoxia induced miR-17-5p expression. Overexpression of miR-17-5p by transfection with miR-17-5p enhanced cell proliferation and migration in normoxia, whereas knockdown of miR-17-5p with anti-miR-17-5p inhibitors significantly reduced cell proliferation and migration. The expression of miR-17-5p target genes, specifically phosphatase and tensin homologue (PTEN) and cyclin-dependent kinase inhibitor 1 (p21WAF1/Cip1, p21), was reduced under moderate hypoxia in PASMCs. Under normoxia, overexpression of miR-17-5p in PASMCs reduced the expression of PTEN and p21. CONCLUSION: Our data indicate that miR-17-5p might play a significant role in hypoxia-induced pulmonary vascular smooth muscle cell proliferation by regulating multiple gene targets, including PTEN and p21, and that miR-17-5p could be a novel therapeutic target for the management of hypoxia-induced PH.
Assuntos
Proliferação de Células/fisiologia , Inibidor de Quinase Dependente de Ciclina p21/fisiologia , MicroRNAs/fisiologia , Miócitos de Músculo Liso/metabolismo , PTEN Fosfo-Hidrolase/biossíntese , Artéria Pulmonar/metabolismo , Hipóxia Celular/fisiologia , Movimento Celular/fisiologia , Células Cultivadas , Inibidor de Quinase Dependente de Ciclina p21/antagonistas & inibidores , Humanos , MicroRNAs/antagonistas & inibidores , Músculo Liso Vascular/citologia , Músculo Liso Vascular/metabolismo , Artéria Pulmonar/citologia , Regulação para Cima/fisiologiaRESUMO
PURPOSE: To investigate the effects of hyaluronic acid (HA) on the inflammation of corneal fibroblasts induced by lipopolysaccharide (LPS). METHODS: Primary rabbit corneal keratocytes were isolated with collagenase. The keratocytes were cultured in a serum-containing medium to induce corneal fibroblasts, which represented the wound repair phenotype of corneal keratocytes. Corneal fibroblasts were treated with LPS with or without 4-methylumbelliferone (4-MU) / high molecular weight hyaluronic acid (HMWHA). The gene expression was evaluated via real-time PCR, immunofluorescence, and western blot. The release of inflammatory cytokines and HA was determined by ELISA. RESULTS: Three types of hyaluronan synthase (HAS) were detected in corneal fibroblasts. LPS stimulation caused the up-regulation of HAS1 and HAS2 expression in corneal fibroblasts. LPS-induced HAS2 expression was significantly inhibited by 4-MU, and accompanied by decreased HA release by the corneal fibroblasts. In the corneal fibroblasts, 4-MU reduced the LPS-stimulated up-regulation of inflammatory cytokines including IL-1, IL-6, IL-8, TNF-α, and also attenuated the LPS-induced up-regulation of inflammatory related receptors including TLR2, TLR4, CD44, and CXCR1. HMWHA treatment resulted in a significant decline in the expression of IL-6, IL-8, TLR4, and CXCR1 responded to LPS stimulation. Consistent with mRNA expression of level, the up-regulation of the release of IL-6 and IL-8 induced by LPS in corneal fibroblasts was significantly attenuated by 4-MU and HMWHA. The LPS-induced expression of IL-8 and its receptor CXCR1 at both the mRNA and protein level were significantly attenuated by 4-MU and HMWHA. CONCLUSION: The inhibitor of HA synthesis 4-MU, and HMWHA successfully reduced LPS-induced inflammation in corneal fibroblasts. The mechanism might be via the inhibition of LPS-induced TLR4 up-regulation.
Assuntos
Ceratócitos da Córnea/patologia , Substância Própria/patologia , Ácido Hialurônico/farmacologia , Ceratite/tratamento farmacológico , Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/farmacologia , Animais , Western Blotting , Células Cultivadas , Ceratócitos da Córnea/efeitos dos fármacos , Ceratócitos da Córnea/metabolismo , Substância Própria/efeitos dos fármacos , Substância Própria/metabolismo , Citocinas/biossíntese , Citocinas/genética , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Regulação da Expressão Gênica/efeitos dos fármacos , Ácido Hialurônico/química , Imuno-Histoquímica , Ceratite/metabolismo , Ceratite/patologia , Lipopolissacarídeos/toxicidade , Peso Molecular , RNA/genética , Coelhos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Purpose: Corneal injury (CI) resulting in corneal opacity remains a clinical challenge. Exosomes (Exos) derived from bone marrow mesenchymal stem cells (BMSCs) have been proven effective in repairing various tissue injuries and are also considered excellent drug carriers due to their biological properties. Recently, microRNA-29b (miR-29b) was found to play an important role in the autophagy regulation which correlates with cell inflammation and fibrosis. However, the effects of miR-29b and autophagy on CI remain unclear. To find better treatments for CI, we used Exos to carry miR-29b and investigated its effects in the treatment of CI. Methods: BMSCs were transfected with miR-29b-3p agomir/antagomir and negative controls (NCs) to obtain Exos-29b-ago, Exos-29b-anta, and Exos-NC. C57BL/6J mice that underwent CI surgeries were treated with Exos-29b-ago, Exos-29b-anta, Exos-NC, or PBS. The autophagy, inflammation, and fibrosis of the cornea were estimated by slit-lamp, hematoxylin and eosin (H&E) staining, immunofluorescence, RTâqPCR, and Western blot. The effects of miR-29b-3p on autophagy and inflammation in immortalized human corneal epithelial cells (iHCECs) were also investigated. Results: Compared to PBS, Exos-29b-ago, Exos-29b-anta, and Exos-NC all could ameliorate corneal inflammation and fibrosis. However, Exos-29b-ago, which accumulated a large amount of miR-29b-3p, exerted excellent potency via autophagy activation by inhibiting the PI3K/AKT/mTOR pathway and further inhibited corneal inflammation via the mTOR/NF-κB/IL-1ß pathway. After Exos-29b-ago treatment, the expressions of collagen type III, α-smooth muscle actin, fibronectin, and vimentin were significantly decreased than in other groups. In addition, overexpression of miR-29b-3p prevented iHCECs from autophagy impairment and inflammatory injury. Conclusions: Exos from BMSCs carrying miR-29b-3p can significantly improve the therapeutic effect on CI via activating autophagy and further inhibiting corneal inflammation and fibrosis.
Assuntos
Autofagia , Lesões da Córnea , Modelos Animais de Doenças , Exossomos , Células-Tronco Mesenquimais , Camundongos Endogâmicos C57BL , MicroRNAs , Animais , MicroRNAs/genética , Exossomos/metabolismo , Exossomos/transplante , Células-Tronco Mesenquimais/metabolismo , Camundongos , Lesões da Córnea/metabolismo , Lesões da Córnea/genética , Lesões da Córnea/terapia , Portadores de Fármacos , Inflamação/metabolismo , Masculino , Células Cultivadas , Humanos , Western BlottingRESUMO
Purpose: This study aimed to explore the effects of elevated KDM4D expression and potential therapeutic effects of Lycium barbarum polysaccharide (LBP) on pterygium. Methods: The expression levels of KDM4D in the primary pterygium (n = 29) and normal conjunctiva (n = 14) were detected by immunohistochemistry. The effects of KDM4D on pterygium fibroblasts were detected by the CCK-8 assay, liquid chromatography-mass spectrometry assay, flow cytometry, and scratch wound healing assay. The relative expression of KDM4D in pterygium fibroblasts stimulated by interleukin (IL)-1ß, IL-6, IL-8, and LBP was detected by quantitative real-time PCR and Western blot. The effects of LBP on pterygium fibroblasts were detected using flow cytometry and scratch wound healing assays. Results: The expression level of KDM4D in pterygium was higher than that in normal conjunctiva. KDM4D increased the cell viability of pterygium fibroblasts. The differentially expressed genes identified in the LM-MS assay enriched in "actin filament organization" and "apoptosis." KDM4D promoted migration and inhibited apoptosis of pterygium fibroblasts in vitro. Inflammatory cytokines, including IL-1ß, IL-6, and IL-8, enhanced the expression of KDM4D in pterygium fibroblasts. LBP inhibited the expression of KDM4D in pterygium fibroblasts and decreased their cell viability. Moreover, LBP attenuated the KDM4D effects on migration and apoptosis of pterygium fibroblasts. Conclusions: Elevated KDM4D expression is a risk factor for pterygium formation. LBP inhibits the expression of KDM4D in pterygium fibroblasts and may be a potential drug for delaying pterygium development.
Assuntos
Túnica Conjuntiva/anormalidades , Medicamentos de Ervas Chinesas , Pterígio , Humanos , Pterígio/tratamento farmacológico , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/metabolismo , Histona Desmetilases com o Domínio Jumonji/metabolismoRESUMO
BACKGROUND: Mutations in the KIF21A gene are detected in the patients with congenital fibrosis of the extraocular muscles. Mutations in the PAX6 gene are detected in the patients with congenital aniridia. CASE PRESENTATION: Herein we report a boy with both congenital fibrosis of extraocular muscles and aniridia. Sequence analysis of his KIF21A and PAX6 genes reveals a 1-bp deletion (c.745delC) in the PAX6 gene and a missense mutation of c.2860C > T (p.Arg954Trp) in KIF21A. CONCLUSIONS: This study demonstrates that the occurrence of independent mutations in more than a single gene in a patient may lead to a complex phenotype.
Assuntos
Aniridia/genética , Proteínas do Olho/genética , Deleção de Genes , Proteínas de Homeodomínio/genética , Cinesinas/genética , Músculos Oculomotores/patologia , Fatores de Transcrição Box Pareados/genética , Proteínas Repressoras/genética , Aniridia/patologia , Criança , Éxons , Fibrose , Humanos , Masculino , Mutação de Sentido Incorreto , Fator de Transcrição PAX6 , Linhagem , FenótipoRESUMO
PURPOSE: To determine the genetic origin of disease in four Chinese families with blepharophimosis syndrome. METHODS: Four Han Chinese families with blepharophimosis syndrome were ascertained and patients underwent complete physical and ophthalmic examinations. Blood samples were collected and genomic DNA was extracted. Sequence analysis of the forkhead transcriptional factor 2 (FOXL2) gene was performed by direct sequencing and mutations were analyzed. RESULTS: Three mutations in FOXL2 were found in four families, including c.672_701dup30 (p.Ala224_Ala234dup10), c.313C>A (p.N105H), and c.430G>T (p.R144W). The c.672_701dup30 (p.Ala224_Ala234dup10) mutation was reported previously and predicted to result in expansions of the polyalanine tract. The mutations of c.313C>A (p. N105H) and c.430G>T (p.R144W) are two novel missense mutations. CONCLUSIONS: Our study further supports the view that the expansion of the polyalanine tract is the hotspot of mutations within FOXL2. The two novel missense mutations detected in this study will expand the mutation spectrum of the FOXL2 gene and contribute to the research on the molecular pathogenesis of FOXL2.
Assuntos
Povo Asiático/genética , Blefarofimose/genética , Fatores de Transcrição Forkhead/genética , Predisposição Genética para Doença , Mutação/genética , Anormalidades da Pele/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , China , Análise Mutacional de DNA , Feminino , Proteína Forkhead Box L2 , Fatores de Transcrição Forkhead/química , Humanos , Masculino , Dados de Sequência Molecular , Linhagem , Polimorfismo de Nucleotídeo Único/genética , Alinhamento de Sequência , Anormalidades UrogenitaisRESUMO
This research investigated the potential use of 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT) as a biocide in aircraft fuel systems, which is rarely studied due to the unique properties of such systems. The study assessed the effectiveness of CMIT against three microbial isolates using minimum inhibitory concentrations and bacteriostatic tests, and showed that CMIT had good activity against them. Electrochemical studies were conducted to determine the impact of CMIT on the 7B04 aluminum alloy, which demonstrated that CMIT acted as a cathodic inhibitor and exhibited certain levels of short-term and long-term corrosion inhibition effects at concentrations of 100 mg L-1 and 60 mg L-1, respectively. Additionally, the research provided insights into the mechanisms governing microbial problems by studying the reaction of CMIT with glutathione and sulfate. Overall, the study suggested that CMIT may be a useful biocide in aircraft fuel systems and provided important information on its efficacy and mechanism of action.
RESUMO
Inflammatory diseases are key contributors to high mortality globally and adversely affect the quality of life. Current treatments include corticosteroids or nonsteroidal anti-inflammatories that may cause systemic toxicity and biologics that may increase the risk of infection. Composite nanoparticles that bear not only the drug payload but also targeting ligands for delivery to inflammation sites at lowered systemic toxicity are established in the nanomedicine field, but their relatively large size often leads to systemic clearance. Metal-based nanoparticles with intrinsic anti-inflammatory properties represent attractive alternatives. They are not only designed to be compact for crossing biological barriers (with the nanoparticle serving as a dual carrier and drug), but also support label-free tracking of their interactions with cells. The review commences with an outline of the common inflammatory diseases, inflammatory pathways involved, and conventional drug-loaded nanoparticles for anti-inflammation. Next, the review features the emerging applications of self-therapeutic metal-based nanoparticles (e.g., gold, coper oxide, platinum, ceria, and zinc oxide) for managing inflammatory diseases in animals over the past three years, focusing on therapeutic outcomes and anti-inflammatory mechanisms. The review concludes with an outlook on the biodistribution, long-term toxicity, and clinical translation of self-therapeutic metal-based nanoparticles.
RESUMO
Purpose: To identify the molecular background of eyelid sebaceous gland carcinomas (SCs), we conducted the integrated whole-exome sequencing and transcriptome sequencing for eyelid SCs in this study. Methods: The genetic alterations were studied by whole-exome sequencing, and the messenger RNA expression was studied using Oxford Nanopore Technologies (ONT) in five paired fresh eyelid SC tissues and adjacent normal tissues. Integrated analysis of exome and transcriptomic information was conducted for filtering candidate driver genes. Protein-protein interaction (PPI) network of filtered candidate genes was analyzed by STRING. The protein expression was verified by immunohistochemistry in 29 eyelid SCs and 17 compared normal sebaceous gland tissues. Results: The average numbers of pathogenic somatic single-nucleotide variants (SNVs) and indels in eyelid SCs were 75 and 28, respectively. Tumor protein p53 (TP53), zinc finger protein 750 (ZNF750), filaggrin 2 (FLG2), valosin-containing protein (VCP), and zinc finger protein 717 (ZNF717) were recurrent mutated genes. A mean of 844 differentially expressed genes (DEGs) were upregulated, and 1401 DEGs were downregulated in SC samples. The intersection of DEG-based pathways and mutation-based pathways was mainly involved in microbial infection and inflammation, immunodeficiency, cancer, lipid metabolism, and the other pathways. The intersection of DEGs and mutated genes consisted of 55 genes, of which 15 genes formed a PPI network with 4 clusters. The PPI cluster composed of scavenger receptor class B member 1 (SCARB1), peroxisome proliferator-activated receptor γ (PPARG), peroxisome proliferator-activated receptor γ coactivator 1α (PPARGC1A) was involved in cholesterol metabolism. The expression of SCARB1 protein was found to be increased, whereas that of PPARG protein was decreased in eyelid SCs compared to that in the normal sebaceous glands. Conclusions: Increased SCARB1 and decreased PPARG indicated that dysregulation of cholesterol metabolism might be involved in carcinogenesis of eyelid SCs. Translational Relevance: The malfunction in cholesterol metabolism might advance our knowledge of the carcinogenesis of eyelid SCs.
Assuntos
Carcinoma , Neoplasias Palpebrais , Humanos , Transcriptoma/genética , Exoma/genética , Glândulas Sebáceas/metabolismo , Glândulas Sebáceas/patologia , PPAR gama/genética , PPAR gama/metabolismo , Sequenciamento do Exoma , Neoplasias Palpebrais/genética , Neoplasias Palpebrais/metabolismo , Neoplasias Palpebrais/patologia , Pálpebras , Carcinogênese/genética , Carcinogênese/metabolismo , Carcinogênese/patologia , Metabolismo dos Lipídeos/genética , Carcinoma/genética , Carcinoma/metabolismo , Carcinoma/patologia , Colesterol/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismoRESUMO
Tissue engineering (TE) cornea is one of the most potential alternatives to the shortage of corneal donors in cornea transplantation. Sodium alginate (SA) hydrogel is commonly used as scaffold in TE. Herein, we present an approach to construct a composite hydrogel, which with SA fiber skeleton structure for shape retention and gelatin surface modification for water retention. The light transmittance, water retention rate, and swelling rate of hydrogels were characterized, and the tensile mechanical properties were also investigated. Keratinocytes were treated with material extract liquor and the results showed that the gelatin modified SA hydrogel has good cytocompatibility. Furthermore, human corneal stromal fibroblasts (HCSFs) from the lenticules were implanted on the surface of gels, and the SA-gelatin hydrogel significantly improved the adhesion and spreading of HCSFs. Finally, we discussed the improvement and application prospect of the composite hydrogel as cornea equivalents.