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BACKGROUND: The first line treatment for moderate to severe active thyroid associated ophthalmopathy is glucocorticoid pulse therapy, but for patients with contraindications to hormone therapy or hormone resistance, it is urgent to find a suitable treatment plan. AIMS: To find a reliable alternative to hormone pulse therapy for thyroid associated ophthalmopathy by comparing the efficacy with first-line treatment regimens. METHODS: Search PubMed, Ovid, Web of science, Cochrane library, and Clinical Trials.gov for randomized controlled trials on the treatment of thyroid associated ophthalmopathy published as of July 7, 2024. Quality evaluation and Bayesian network analysis were conducted using RevMan 5.3 software, STATA15.0 software, and ADDIS 1.16.8 software. RESULTS: A total of 666 patients were included in 11 studies and 8 interventions. Network analysis showed that the three interventions of mycophenolate mofetil combined with glucocorticoids, Teprotumumab and 99Tc-MDP were superior to glucocorticoid pulse therapy in improving clinical activity scores and proptosis. The regimen of glucocorticoids combined with statins can improve the quality of life score and diplopia score of patients. Neither methotrexate combined with glucocorticoids nor rituximab alone showed additional advantages when compared with glucocorticoid pulse therapy. CONCLUSION: Mycophenolate mofetil combined with glucocorticoid therapy is very beneficial for moderate to severe active thyroid associated ophthalmopathy. Mycophenolate mofetil may be a good choice when patients have contraindications to hormone use or hormone resistance. Teprotumumab is very promising and may be able to avoid patients undergoing orbital decompression surgery. The durability and safety of its long-term efficacy need to be further observed.
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Teorema de Bayes , Glucocorticoides , Oftalmopatia de Graves , Humanos , Oftalmopatia de Graves/tratamento farmacológico , Oftalmopatia de Graves/diagnóstico , Glucocorticoides/uso terapêutico , Glucocorticoides/administração & dosagem , Metanálise em Rede , Qualidade de Vida , Anticorpos Monoclonais HumanizadosRESUMO
Objective To analyze the serological characteristics and clinical significance of IgG anti-D and anti-C combined antibodies and anti-E and anti-c combined antibodies in patients negative for RhDC and RhEc antigens. Methods The clinical data and laboratory results of 12 cases with two types of irregular antibodies were recorded and analyzed, including age, sex, history of blood transfusion/pregnancy, ABO and RhD blood group identification, Rh antigen typing, irregular antibody screening, antibody-specific identification, absorption-elution tests, antibody titer determination and cross-matching tests. Results Among the 12 patients, the mean age was 51.4±16.9 years. Nine patients had a history of blood transfusion; eight patients had a history of pregnancy; five patients had both. Serological tests showed positive antibody screening and incompatible cross-matching. The results of antibody-specific identification and absorption-elution tests showed the presence of both IgG anti-D and anti-C antibodies in three patients, with anti-D titers at 16-32, and anti-C titers at 8-16. Nine patients had both IgG anti-E and anti-c antibodies, with the titers of anti-E and anti-c antibodies at 8-16. From the patients with combined anti-D and anti-C antibodies, suspended red blood cells of ABO identical type, RhD negative and other Rh antigens as ccee were selected. From patients with combined anti-E and anti-c antibodies, suspended red blood cells of ABO identical type, RhD positive and other Rh antigens as CCee were selected. Cross-matching blood test results showed no agglutination or hemolysis in saline, polycoagulant and anti-human globulin media. Conclusion Blood transfusion and/or pregnancy are the primary causes of irregular antibodies in two Rh systems, leading to positive antibody screening and cross-match incompatibility. Routine compatibility transfusion of Rh antigens, based on ABO homotypic blood transfusion, is of great value and significance for the safety of clinical blood transfusion and the prevention of hemolytic disease of the newborn.
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Incompatibilidade de Grupos Sanguíneos , Tipagem e Reações Cruzadas Sanguíneas , Isoanticorpos , Sistema do Grupo Sanguíneo Rh-Hr , Humanos , Sistema do Grupo Sanguíneo Rh-Hr/imunologia , Feminino , Pessoa de Meia-Idade , Masculino , Adulto , Incompatibilidade de Grupos Sanguíneos/imunologia , Isoanticorpos/sangue , Isoanticorpos/imunologia , Idoso , Gravidez , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Sistema ABO de Grupos Sanguíneos/imunologia , Transfusão de SangueRESUMO
A critical shortage of donor corneas exists worldwide. Hydrogel patches with a biological architecture and functions that simulate those of native corneas have garnered considerable attention. This study introduces a stromal structure replicating corneal patch (SRCP) composed of a decellularized cornea-templated nanotubular skeleton, recombinant human collagen, and methacrylated gelatin, exhibiting a similar ultrastructure and transmittance (above 80 %) to natural cornea. The SRCP is superior to the conventional recombinant human collagen patch in terms of biomechanical properties and resistance to enzymatic degradation. Additionally, SRCP promotes corneal epithelial and stromal cell migration while preventing the trans-differentiation of stromal cells into myofibroblasts. When applied to an ocular surface (37 °C), SRCP releases methacrylated gelatin, which robustly binds SRCP to the corneal stroma after activation by 405 nm light. Compared to gelatin-based photocurable hydrogel, the SRCP better supports the restoration of normal corneal curvature and withstands deformation under an elevated intraocular pressure (100 mmHg). In an in vivo deep anterior-corneal defect model, SRCP facilitated epithelial healing and vision recovery within 2 weeks, maintained graft structural stability, and inhibited stromal scarring at 4 weeks post-operation. The ideal performance of the SRCP makes it a promising humanized corneal equivalent for sutureless clinical applications.
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OBJECTIVES: To investigate the ultrasound (US) characteristics of metastatic malignancies (MM) in the major salivary glands and to assess the diagnostic value of the close relationship with the glandular capsule in identifying MM. METHODS: From January 2016 and April 2022, 122 patients with major salivary gland malignancies, including 20 patients with MM and 102 patients with primary malignancies (PM) confirmed by histopathological examination, were enrolled in this study. Their clinicopathologic and US data were recorded and analyzed. The diagnostic performance of the close relationship with the glandular capsule for differentiating MM from PM was analyzed. RESULTS: The mean age of MM were older than that of PM (59.50 ± 14.57 vs. 49.96 ± 15.73, p = 0.013). Compared with PM patients, MM were associated with a higher prevalence of local pain symptoms (p = 0.007) and abnormal facial nerve function (p < 0.001). MM were also more frequently characterized by unclear borders, rough margins, irregular shapes, heterogeneous internal echos, absence of cystic areas, presence of calcifications, close relationship with the glandular capsule, and US-reported positive cervical lymph nodes (all p < 0.05). The close relationship with the glandular capsule showed to be a good indicator in distinguishing between MM and PM, with an area under the receiver operating characteristic curve of 0.863, a sensitivity of 100%, a specificity of 72.5%, and an accuracy of 92.2%. Positive and negative predictive were calculated at 41.7% and 100%, respectively. CONCLUSIONS: The US finding of a close relationship with the glandular capsule is a highly sensitive diagnostic indicator for MM. Following this finding, US-guided needle biopsy should be recommended to further confirm the diagnosis.
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Keratomycosis, caused by pathogenic fungi, is an intractable blinding eye disease. Corneal penetration is an essential requirement for conventional antifungal medications to address keratomycosis. Due to the distinctive anatomical and physiological structure of the cornea, the therapeutic efficacy is hampered by the inadequate penetration capacity. Despite the emergence of diverse antifungal drug delivery systems and advanced antifungal nanomaterials, it has remained challenging to achieve corneal penetration over the past decade. This study fabricates a penetrative ionic organic molecular cage-based nanozyme (OMCzyme) for treating keratomycosis. The synthesis of OMCzyme involved two steps. Initially, the ionic OMC is synthesized by a [2+3] cycloimination reaction of triformylphloroglucinol and 2,3-diaminopropionic acid. Subsequently, OMCzyme is fabricated by coordination of Fe2⺠with carboxyl anions and phenolic hydroxyls in the organic cage, and further deposition of silver nanoparticles on the surface of OMC-Fe complex. The as-prepared OMCzyme demonstrates excellent water dispersion, peroxidase-like activity, in vitro and in vivo biocompatibility, and corneal penetration. Notably, the nanozyme displays targeted antifungal activity, effectively combating Fusarium solani with negligible cytotoxicity toward human corneal epithelial cells. The hybrid mimic is further demonstrated to be effective in treating keratomycosis in mice, indicating the potential of OMCzyme for curing fungal infectious diseases.
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Purpose: This study aims to elucidate the calcitonin gene-related peptide (CGRP) mediation and primary mechanism of corneal sensory nerves on tear production of the lacrimal gland. Methods: Mouse corneal denervation models were constructed through surgical axotomy, pharmacologic treatment with capsaicin or resiniferatoxin, and Trpv1-Cre/DTR mice with diphtheria toxin injection. The capsaicin-treated mice received subconjunctival injection of CGRP or substance P, while the normal C57BL/6J mice were administered with CGRP receptor antagonist BIBN-4096. Furthermore, double immunostaining of c-FOS+ and choline acetyltransferase was used to evaluate the activation of the superior salivatory nucleus (SSN). Mouse lacrimal glands were collected for transcriptomic sequencing and subsequent RNA and protein expression analysis. Results: The corneal denervated mice exhibited a significant reduction in corneal sensitivity and tear secretion. In capsaicin-treated mice, tear secretion decreased to 2.5 ± 0.5 mm compared to 6.3 ± 0.9 mm in control mice (P < 0.0001). However, exogenous administration of CGRP in capsaicin-treated mice increased tear secretion from 2.6 ± 0.5 mm to 4.5 ± 0.5 mm (P = 0.0009), while BIBN-4096 treatment reduced tear secretion to 3.4 ± 0.5 mm when compared to 7.3 ± 0.7 mm in control mice (P = 0.0022). Furthermore, c-FOS+ cell number in the SSN increased by twofold (P = 0.0168) after CGRP administration compared with capsaicin-treated mice. In addition, the expressions of CCNA2, Ki67, PCNA, and CDK1 in acinar cells of the lacrimal gland were impaired by corneal denervation and alleviated by CGRP administration. Conclusions: CGRP released by corneal sensory nerves mediates tear secretion of the lacrimal gland, providing a new strategy for improving tear secretion in patients with neurotrophic keratitis.
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Peptídeo Relacionado com Gene de Calcitonina , Aparelho Lacrimal , Animais , Camundongos , Capsaicina , Genes fos , Camundongos Endogâmicos C57BL , Proteínas Proto-Oncogênicas c-fosRESUMO
How to evaluate the resilience level and change trend of supply chain is an important research direction in current supply chain management practice. This paper proposes a new method of supply chain resilience assessment based on hesitant fuzzy linguistic term set (HFLTS) and matter element extension theory. Firstly, based on the research status quo at home and abroad, a low-carbon enterprise supply chain resilience assessment index system is established, which includes six first-level indicators and corresponding 21 second-level indicators of product supply resilience, resource resilience, partner resilience, information response resilience, financial resilience and knowledge resilience. Secondly, HFLTS was used to collect expert opinions and Ordered Weighted Arithmetic (OWA) to calculate the expert composite language, by which the fuzzy evaluation matrix of supply chain resilience assessment indicators was obtained. Once again, the resilience indicator weights are determined based on a game-theoretic portfolio assignment method combining the best-worst method (BWM) and the CRITIC method. Finally, the nearness degree function is combined with the extension comprehensive evaluation method to improve the matter element extension model, and the supply chain resilience assessment model of low-carbon enterprises based on the game theory combination assignment-improved matter element extension is established. Taking X low-carbon enterprise as an example, the evaluation results show that the supply chain resilience level of this enterprise is II, and the eigenvalue of the grade variable is 2.69, and the supply chain resilience is shifting to III, and the supply chain resilience is shifting to III, which indicates that the supply chain resilience of this enterprise is being enhanced. Therefore, the improved matter element extension not only ensures the accuracy of the evaluation results, but also has higher prediction accuracy.
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Lógica Fuzzy , Resiliência Psicológica , LinguísticaRESUMO
Neurotrophic keratopathy (NK) is a challenging disease with the reduced innervation to the cornea. To establish a genetic and stable mouse model of NK, we utilized the TRPV1-DTR mice with intraperitoneal injection of diphtheria toxin (DT) to selectively eliminate TRPV1 neurons. After DT administration, the mice exhibited robust ablation of TRPV1 neurons in the trigeminal ganglion, accompanied with reduced corneal sensation and nerve density, as well as the decreased calcitonin-gene-related peptide (CGRP) and substance P levels. According to disease progression of TRPV1 neuronal ablation, tear secretion was reduced from day 3, which followed by corneal epithelial punctate lesions from day 7. From day 11 to day 16, the mice exhibited persistent corneal epithelial defects and stromal edema. By day 21, corneal ulceration and stromal melting were observed with the abundant inflammatory cell infiltration, corneal neovascularization, and enhanced cell apoptosis. Moreover, subconjunctival injection of CGRP delayed the NK progression with the characteristics of reduced severe corneal epithelial lesions and corneal inflammation. In addition, the impairments of conjunctival goblet cells, lacrimal gland, and meibomian gland were identified by the diminished expression of MUC5AC, AQP5, and PPARγ, respectively. Therefore, these results suggest that the TRPV1-DTR mice may serve as a reliable animal model for the research of NK pathogenesis.
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Distrofias Hereditárias da Córnea , Ceratite , Doenças do Nervo Trigêmeo , Camundongos , Animais , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Córnea/metabolismo , Neurônios/metabolismo , Modelos Animais de Doenças , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismoRESUMO
Purpose: To reveal the role of transient receptor potential cation subfamily M member 8 (TRPM8) channels in herpes simplex keratitis (HSK). Methods: HSK models were established using TRPM8 knockout (TRPM8-/-) mice and their wild-type (WT) littermates. The infected corneas were graded and harvested to evaluate the mRNA levels of inflammatory factors through quantitative real-time polymerase chain reaction (RT-PCR), as well as the infiltration of inflammatory cells through immunofluorescence staining and flow cytometry. Viral titers were determined by plaque assay and absolute quantitative method. RNA-sequencing was conducted to elucidate the transcriptome of corneal epithelium in response to TRPM8 knockout after infection. The anti-inflammatory effect of TRPM8 agonist menthol was documented via subconjunctival administration. Results: Compared to their wild-type counterparts, TRPM8-deficient mice exhibited exacerbated infection symptoms and thicker corneas in HSK models. Infection in TRPM8-deficient mice resulted in significant lymphocyte infiltration, primarily consisting of Ly6G+ CD11b+ cells. Additionally, TRPM8-deficient mice displayed increased levels of corneal viral titers after infection, along with decreased expression of interferon-stimulated genes (ISGs). Subconjunctival administration of menthol effectively alleviated infection-induced symptoms and Ly6G+ CD11b+ cell infiltration in herpes simplex virus type 1 (HSV-1)-treated mice. Conclusions: TRPM8 promoted host resistance to HSV-1 infection by suppressing the accumulation of Ly6G+ CD11b+ cells and virus replication. These findings suggest that targeting TRPM8 could be valuable for therapeutic interventions against HSV-1 infections.
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Herpes Simples , Ceratite Herpética , Canais de Cátion TRPM , Animais , Camundongos , Carga Viral , Mentol , Ceratite Herpética/tratamento farmacológico , Córnea , Canais de Cátion TRPM/genéticaRESUMO
Endothelial keratoplasty is the main surgical procedure for treating corneal endothelial dysfunction (CED), which is limited by the global shortage of donor corneas. Herein, we developed and evaluated the modified thermoplastic polyurethane (M-TPU) films with gelatin-glycidyl methacrylate to replace the corneal endothelial function and maintain corneal transparency. The films displayed comparable light transmission characteristics with normal corneas and clinically favorable mechanical properties for surgical manipulation. After surface modification, the hydrophilicity and biocompatibility of M-TPU films were significantly improved. In the rabbit CED model, the M-TPU implants exhibited firm adhesion to the exposed stromal surface. The rabbit corneal transparency and thickness could be restored completely within 1 week of M-TPU film implantation. There was no significant inflammatory reaction and immune rejection during the follow-up of 1 month. Proteomic analysis suggested that the complement inhibition, the increase of mineral absorption, and the decrease of P53 apoptosis signaling pathway and lysine degradation might be beneficial in maintaining the corneal transparency. Overall, our study demonstrated the potential of M-TPU films as artificial implants for the replacement of corneal endothelial function to restore corneal thickness and transparency.
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Poliuretanos , Proteômica , Animais , Coelhos , Endotélio Corneano/cirurgia , Córnea , Próteses e ImplantesRESUMO
OBJECTIVES: Autoimmune hepatitis (AIH) is an aberrant autoimmune condition mediated by T cell abnormality, which may cause fulminant liver failure and persistent liver injury. This study aimed to disclose the histopathological and functional engagement of interleukin (IL)-26, a potent inflammation mediator, in AIH disease progression. METHODS: We conducted immunohistochemical staining on liver biopsy samples to evaluate intrahepatic expression of IL-26. Cellular sources of hepatic IL-26 were detected by confocal microscopy. Flow cytometry was employed to determine the immunological alterations of CD4+ and CD8+ T cells following in vitro IL-26 treatment on primary peripheral blood mononuclear cells from healthy controls. RESULTS: Statistically significant increase in IL-26 level was observed in AIH (n = 48) liver samples in comparison with patients having chronic hepatitis B (n = 25), nonalcoholic fatty liver disease (n = 18), and healthy donors for living donor liver transplantation (n = 10). The number of intrahepatic IL-26+ cells was positively correlated with histological and serological severity. An immunofluorescence staining indicated that liver-infiltrating CD4+ T cells, CD8+ T cells, and CD68+ macrophages orchestrated IL-26 secretion in AIH. Both CD4+ and CD8+ T cells demonstrated effective activation, lytic, and proinflammatory functions upon IL-26 stimulation. CONCLUSION: We observed elevated IL-26 in AIH liver which promoted T cell activation and cytotoxic capacity, indicating a therapeutic potential of IL-26 intervention in AIH.
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Hepatite Autoimune , Transplante de Fígado , Humanos , Linfócitos T CD8-Positivos/metabolismo , Linfócitos T CD8-Positivos/patologia , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/patologia , Fígado/patologia , Doadores VivosRESUMO
Purpose: To reveal the role of cold-sensing transient receptor potential melastatin 8 (TRPM8) channels in corneal epithelial wound healing. Methods: Cold sensitivity, tear production, corneal thickness, and corneal opacity assessments were used to evaluate the effect of Trpm8 knockout on the ocular surface. A corneal epithelial wounding model was generated by scraping the corneal epithelium once or multiple times using C57BL/6J (wild-type [WT]) and Trpm8-/- mice. The processes of corneal epithelial repair and corneal epitheliopathy were observed and recorded. Corneas were collected for sequencing, immunofluorescence staining, hematoxylin and eosin staining, and quantitative PCR. Results: The perception of coldness, basal tear secretion, and corneal thickness were decreased in young Trpm8-/- mice compared with those in WT mice, except for the corneal sensitivity. Corneal opacity and increased corneal thickness were observed in aged Trpm8-/- mice. TRPM8 deficiency promoted corneal epithelial wound closure, consistent with the observed increase in Ki67-positive epithelial cells, and the pharmacological activation of TRPM8 in WT mice delayed corneal re-epithelization. After subjecting mice to multiple injuries, squamous metaplasia emerged in Trpm8-/- corneas, as verified by cytokeratin-1 and small proline-rich protein 1B-positive staining. The IFN-ß and IFN-γ signaling pathways were significantly activated in Trpm8-/- mice, which was confirmed based on the up-regulated expression of the key mediators, signal transducer and activator of transcription-1 and phosphor-signal transducer and activator of transcription-1, as well as the induction of IFN-stimulated genes, compared with levels in WT mice. Conclusions: In corneal wound healing, the loss of TRPM8 function could promote epithelial repair, but predispose the cornea to epithelial lesions.
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Lesões da Córnea , Opacidade da Córnea , Epitélio Corneano , Canais de Cátion TRPM , Camundongos , Animais , Camundongos Endogâmicos C57BL , Lesões da Córnea/metabolismo , Córnea/metabolismo , Epitélio Corneano/metabolismo , Cicatrização/fisiologia , Opacidade da Córnea/metabolismo , Canais de Cátion TRPM/genética , Canais de Cátion TRPM/metabolismoRESUMO
Dry eye is a multifactorial disease that causes dryness, inflammation and damage of ocular surface. Subcutaneous injection of the muscarinic cholinergic antagonist scopolamine under desiccating stress reduces tear production and induces dry eye symptoms in mice. However, the expression profile and pathogenic changes of the lacrimal gland remain incompletely understood. In the present study, we performed comparative transcriptomic analysis of lacrimal glands from the control and scopolamine-treated mice. Primary analysis identified 677 upregulated genes and 269 downregulated genes in the lacrimal gland of mice with scopolamine treatment. Unexpectedly, KEGG pathway and hierarchical clustering analysis showed the enrichment of "DNA replication" and "cell cycle" categories in the upregulated genes. Subsequently, we confirmed that the acinar cells were the major proliferating cells of lacrimal gland, which exhibited significant increasing of the proliferating cell nuclear antigen (PCNA) expression after scopolamine treatment, accompanied with the upregulation of DNA damage marker γ-H2AX. More importantly, both prophylactic and therapeutic administration of the cyclin-dependent kinase (CDK) inhibitor AT7519 rescued the tear reduction and alleviated dry eye severity in the scopolamine-treated mice, including corneal epithelial barrier function, lacrimal and corneal inflammation, and conjunctival goblet cell density. Therefore, we conclude that aberrant acinar cell proliferation is involved in the scopolamine-induced tear reduction and dry eye onset, which can be improved by AT7519 treatment.
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Síndromes do Olho Seco , Aparelho Lacrimal , Camundongos , Animais , Escopolamina/toxicidade , Síndromes do Olho Seco/metabolismo , Aparelho Lacrimal/metabolismo , Lágrimas/metabolismo , Proliferação de Células , Inflamação/metabolismo , Modelos Animais de DoençasRESUMO
Corneal transparency and integrity are essential for obtaining good vision; nevertheless, squamous metaplasia (SQM) of ocular epithelium is a kind of serious blinding corneal diseases, without therapeutic medication in clinic. Here, we found that deficiency of the autoimmune regulator (AIRE) in corneas spontaneously developed corneal plaques. Using corneal abrasion model, we revealed that deletion of Aire not only resulted in delayed corneal re-epithelialization, but also promoted a cell-fate transition from transparent corneal epithelium to keratinized epithelium, histopathologically characterized with SQM based on the transcriptomic analysis. Mechanistically, Aire-deficient corneas led to the heightened Type I interferon (IFN-I)/STAT1 signaling after abrasion. Pharmacological blockade of IFN-I/JAK/STAT1 signaling in Aire-knockout (KO) corneas not only accelerated epithelial wound healing, but also alleviated corneal plaques and SQM. Collectively, our findings revealed critical roles of AIRE in governing corneal epithelial homeostasis and pathologic keratinization, and further identified IFN-I/STAT1 signaling as a potential target for treating ocular surface diseases with SQM, and even for treating pathological scenarios related to SQM in other tissues.
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Carcinoma de Células Escamosas , Epitélio Corneano , Interferon Tipo I , Camundongos , Animais , Córnea/patologia , Epitélio Corneano/patologia , Metaplasia/patologia , Fator de Transcrição STAT1/genéticaRESUMO
AIM: To investigate the influence of dexmedetomidine (Dex) on cerebral ischemia/reperfusion (I/R)-injured rat neuronal cells by regulating the Sphk1/S1P pathway. METHODS: The rats were divided into the following groups, with 18 rats in each group categorized on the basis of random number tables: sham (Sham), I/R (I/R), Dex, Sphk1 inhibitor (PF-543), and Dex together with the Sphk1 agonist phorbol-12-myristate-13-acetate (Dex+PMA). The neurological functions of the rats were assessed by the Longa scoring system at 24 h post reperfusion. The area of brain infarction was inspected using 2,3,5-triphenyltetrazolium chloride staining, and the water content of brain tissue was determined by the dry-wet weight method. The morphology of neurons in the CA1 region of the rat hippocampus was inspected using Nissl staining, while the apoptosis of neurons in this region was detected by terminal-deoxynucleotidyl transferase mediated nick end labeling staining. The Sphk1 and S1P protein levels were determined by immunofluorescence and western blotting, respectively. RESULTS: Compared to the I/R group, rats in the Dex, PF-543, and Dex+PMA groups had a significantly lower neurological function score, as well as lower brain water content and a decreased infarction area. Moreover, the apoptotic index of the neurons and the Sphk1 and S1P levels in the hippocampal CA1 region were significantly lower in these groups (p<0.05). PMA, an agonist of Sphk1, was able to reverse the protective effects of Dex on I/R-induced neuronal cell injury. CONCLUSION: Dex could protect cerebral I/R-induced neuronal cell injury by suppressing the Sphk1/S1P signaling pathway.
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Isquemia Encefálica , Dexmedetomidina , Traumatismo por Reperfusão , Animais , Ratos , Apoptose , Isquemia Encefálica/tratamento farmacológico , Isquemia Encefálica/metabolismo , Região CA1 Hipocampal/metabolismo , Infarto Cerebral , Dexmedetomidina/farmacologia , Neurônios/metabolismo , Ratos Sprague-Dawley , Reperfusão , Traumatismo por Reperfusão/prevenção & controle , Traumatismo por Reperfusão/metabolismo , Transdução de SinaisRESUMO
BACKGROUND: Glycosylation exhibits crucial effect on hepatocellular carcinoma (HCC) progression. Long non-coding RNAs (lncRNAs) are involved in multilevel regulation of gene transcription during tumor development. The purpose of this study is to clarify the potential mechanism that HOTAIR modulates hepatocellular carcinoma progression by activating FUT8/core-fucosylated Hsp90/MUC1/STAT3 feedback loop via JAK1/STAT3 cascade. METHODS: qRT-PCR was used to show the differential expression of genes. Functional experiments were used to measure the malignancy of HCC cells. ChIP and co-IP assays showed the directly interaction of the key molecules. Xenografts was conducted to show the in vivo effects. RESULTS: Upregulation of FUT8 showed closely correlation with HCC progression. Core-fucosylation of Hsp90 stabilized MUC1 binding to the downstream p-STAT3, which involved in the activation of JAK1/STAT3 cascade. STAT3 was identified as the regulator of FUT8 and MUC1 transcription, while FUT8 and MUC1 impacted STAT3 level both in nuclear and cytoplasm. HOTAIR recruited P300 to efficiently bind with STAT3. The transcript complex co-modulated the transcrption of FUT8 and MUC1. Moreover, highly HOTAIR expression also exhibited closely correlation with HCC progression. CONCLUSIONS: FUT8 triggered core-fucosylated-Hsp90/MUC1/P300-HOTAIR-STAT3 cascade via JAK1/STAT3 pathway, which exhibited as positive feedback loop during HCC progression.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , RNA Longo não Codificante , Humanos , Carcinoma Hepatocelular/patologia , Glicosilação , Neoplasias Hepáticas/patologia , Retroalimentação , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Proliferação de Células , Janus Quinase 1/genética , Janus Quinase 1/metabolismo , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Mucina-1/genética , Mucina-1/metabolismoRESUMO
Myocardial infarction (MI) is one of the important factors leading to death in today's society. Therefore, to study the related mechanism of MI and reduce myocardial ischemia-reperfusion injury is an important link to reduce MI injury. MI mice in vivo and cell model in vitro were constructed. The cardiac function and MI area of mice were detected, and myocardial tissue injury was detected by HE staining. ALAS2 expression in mice myocardial tissue was detected by IHC. The expressions of lncRNA-SNHG8, METTL3, PTBP1 and ALAS2 in myocardial tissue or cardiomyocytes were detected by qRT-PCR assay. MTT assay was used to measured viability of cardiomyocytes. The oxidative stress level in myocardial tissue or cardiomyocytes was detected by ELISA assay and ROS assay. RIP-qPCR and RNA pulldown assays determined the interaction between METTL3 and lncRNA-SNHG8, as well as PTBP1 and ALAS2. lncRNA-SNHG8 knockdown in MI mice was reduced myocardial infarction size, alleviated myocardial tissue injury and oxidative stress, and inhibited ALAS2 expression in myocardial tissue. RNA pulldown and RIP assays showed that lncRNA-SNHG8 binged with PTBP1 and PTBP1 interacted with ALAS2 mRNA. Knockdown of lncRNA-SNHG8, METTL3 or PTBP1 in MI cells enhanced viability of myocardial cells, attenuated ROS release and MDA level, increased SOD level, alleviated oxidative stress. ALAS overexpression attenuated the corresponding effect of knockdown of lncRNA-SNHG8 and/or PTBP1 on MI cells. In sum, our paper is demonstrated for the first time that METTL3 can promote lncRNA-SNHG8 through m6A modification, thereby regulating ALAS2 to induce oxidative stress and aggravate myocardial injury.
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MicroRNAs , Infarto do Miocárdio , RNA Longo não Codificante , Camundongos , Animais , MicroRNAs/genética , RNA Longo não Codificante/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Infarto do Miocárdio/metabolismo , Estresse Oxidativo , Ribonucleoproteínas Nucleares Heterogêneas/metabolismo , Proteína de Ligação a Regiões Ricas em Polipirimidinas/genética , Proteína de Ligação a Regiões Ricas em Polipirimidinas/metabolismo , Metiltransferases/metabolismoRESUMO
The present study was aimed to investigate the role and mechanism of glutaminolysis of cardiac fibroblasts (CFs) in hypertension-induced myocardial fibrosis. C57BL/6J mice were administered with a chronic infusion of angiotensin II (Ang II, 1.6 mg/kg per d) with a micro-osmotic pump to induce myocardial fibrosis. Masson staining was used to evaluate myocardial fibrosis. The mice were intraperitoneally injected with BPTES (12.5 mg/kg), a glutaminase 1 (GLS1)-specific inhibitor, to inhibit glutaminolysis simultaneously. Immunohistochemistry and Western blot were used to detect protein expression levels of GLS1, Collagen I and Collagen III in cardiac tissue. Neonatal Sprague-Dawley (SD) rat CFs were treated with 4 mmol/L glutamine (Gln) or BPTES (5 μmol/L) with or without Ang II (0.4 μmol/L) stimulation. The CFs were also treated with 2 mmol/L α-ketoglutarate (α-KG) under the stimulation of Ang II and BPTES. Wound healing test and CCK-8 were used to detect CFs migration and proliferation respectively. RT-qPCR and Western blot were used to detect mRNA and protein expression levels of GLS1, Collagen I and Collagen III. The results showed that blood pressure, heart weight and myocardial fibrosis were increased in Ang II-treated mice, and GLS1 expression in cardiac tissue was also significantly up-regulated. Gln significantly promoted the proliferation, migration, mRNA and protein expression of GLS1, Collagen I and Collagen III in the CFs with or without Ang II stimulation, whereas BPTES significantly decreased the above indices in the CFs. α-KG supplementation reversed the inhibitory effect of BPTES on the CFs under Ang II stimulation. Furthermore, in vivo intraperitoneal injection of BPTES alleviated cardiac fibrosis of Ang II-treated mice. In conclusion, glutaminolysis plays an important role in the process of cardiac fibrosis induced by Ang II. Targeted inhibition of glutaminolysis may be a new strategy for the treatment of myocardial fibrosis.
Assuntos
Ratos , Camundongos , Animais , Ratos Sprague-Dawley , Angiotensina II/farmacologia , Fibroblastos , Camundongos Endogâmicos C57BL , Fibrose , Colágeno/farmacologia , Colágeno Tipo I/metabolismo , RNA Mensageiro/metabolismo , Miocárdio/patologiaRESUMO
Objective: To understand the knowledge, attitude, and current status of vaccination of herpes zoster vaccination among urban residents aged 25 years and above in China. Methods: In August to October 2022, a convenience sampling method was used to survey residents aged 25 years and above at 36 community centers in 9 cities across China. Questionnaires were used to collect basic information, knowledge, and attitude toward herpes zoster and its vaccination, as well as vaccination status and reasons for non-vaccination among residents. Results: A total of 2 864 urban residents were included in the study. The total score of residents' cognition of herpes zoster and its vaccine was 3.01±2.08, and the total score of their attitude was 18.25±2.76. Factors such as being male (β=-0.45, P<0.001), older than 40-59 years (β=-0.34, P=0.023) or ≥60 years (β=-0.68, P<0.001), married (β=-0.69, P=0.002) were negatively associated with knowledge score. The educational level of high school or secondary school (β=0.44, P=0.036), college (β=0.65, P=0.006), bachelor's degree and above (β=1.20, P<0.001), annual net household income ≥120 000 Yuan in 2021 (β=0.42, P=0.020), having urban employee medical insurance (β=0.62, P=0.030), having public or commercial medical insurance (β=0.65, P=0.033), and having a history of chickenpox (β=0.29, P=0.025) were positively associated with knowledge scores. Being male (β=-0.38, P=0.008) and not remembering a history of chickenpox (β=-0.49, P=0.012) were negatively associated with attitude scores. Annual net household income in 2021 was between 40 000-80 000 Yuan (β=0.44, P=0.032) or between 80 000-120 000 Yuan (β=0.62, P=0.002) or ≥120 000 Yuan (β=0.93, P<0.001), and a history of herpes zoster (β=0.59, P=0.004) were positively associated with attitude scores. Of the 2 864 residents surveyed, only 29 (1.01%) had received the herpes zoster vaccine, with a vaccination rate of 1.70% for those aged 50 years and above, with the main reason for non-vaccination being lack of knowledge about the herpes zoster vaccine, followed by the high price. 42.67% of the population said they would consider getting the herpes zoster vaccine in the future. Conclusion: Low knowledge of herpes zoster and its vaccine, positive attitudes towards the preventive effects of herpes zoster and its vaccine, and extremely low vaccination rates among the urban population in China call for multiple measures to strengthen health education and vaccination recommendations for residents, especially for the elderly, low-education and low-income populations.