Abstract: The concept of higher education is the accumulation and philosophical implication of China’s excellent traditional culture, as well as the humanistic spirit and philosophical thought with Chinese characteristics. However, the current concept of higher education has problems, such as inaccurate target positioning, emphasizing theoretical teaching, neglecting practice and being difficult to adapt to market and social needs. How to solve these problems and promote the concept of higher education to glow with new vitality in the new era is currently an urgent issue to be solved. Therefore, this article focused on traditional Chinese philosophical culture and explored the interactive relationship between traditional Chinese philosophical culture and higher education concepts. This article attempted to analyze the relationship between traditional Chinese philosophical culture and higher education concepts from the perspective of their interaction, significance and ways to achieve interaction. The study found that the interaction between traditional Chinese philosophical culture and higher education concepts is of great significance, and it was an important starting point for educational reform to demonstrate new achievements in the new era. This article hoped that the study of the interactive relationship between the two can help people better understand the concept of higher education, better grasp the significance of higher education concepts in the new era from both knowledge and behavior, and provide more theoretical basis for higher education reform. Abstract Chinas China s characteristics However positioning teaching needs solved Therefore behavior

Resumo: O conceito de ensino superior é o acúmulo e a implicação filosófica da excelente cultura tradicional da China, bem como o espírito humanístico e o pensamento filosófico com características chinesas. No entanto, o conceito atual de ensino superior tem problemas, como o posicionamento de metas imprecisas, a ênfase no ensino teórico, a negligência da prática e a dificuldade de adaptação às necessidades sociais e do mercado. Como resolver esses problemas e promover o conceito de ensino superior, para que brilhe com nova vitalidade, na nova era, é uma questão urgente a ser resolvida neste momento. Portanto, este artigo se concentrou na cultura filosófica tradicional chinesa e explorou a relação interativa entre a cultura filosófica tradicional chinesa e os conceitos de ensino superior. O texto procurou analisar a relação entre a cultura filosófica tradicional chinesa e os conceitos de ensino superior, a partir da perspectiva de sua interação, importância e formas de alcançar a interação. O estudo constatou que a interação entre a cultura filosófica tradicional chinesa e os conceitos de ensino superior é de grande importância, e foi um importante ponto de partida para a reforma educacional demonstrar novas conquistas, na nova era. Com este artigo, espera-se que o estudo da relação interativa entre ambos possa ajudar as pessoas a entenderem melhor o conceito de ensino superior, compreender melhor a importância dos conceitos de ensino superior, na nova era, tanto do ponto de vista do conhecimento quanto do comportamento, e fornecer mais base teórica para a reforma do ensino superior. Resumo China chinesas entanto imprecisas teórico mercado vitalidade era momento Portanto conquistas esperase espera comportamento

Abstract Objectives Myocardial Infarction (MI) is the leading cause of chronic heart failure. Previous studies have suggested that Vav3, a receptor protein tyrosine kinase signal transducer, is associated with a variety of cellular signaling processes such as cell morphology regulation and cell transformation with oncogenic activity. However, the mechanism of Vav3-mediated MI development requires further investigation. Method Here, The authors established an MI rat model by ligating the anterior descending branch of the left coronary artery, and an MI cell model by treating cardiomyocytes with H2O2. Microarray analysis was conducted to identify genes with differential expression in heart tissues relevant to MI occurrence and development. Vav3 was thus selected for further investigation. Results Vav3 downregulation was observed in MI heart tissue and H2O2-treated cardiomyocytes. Administration of Lentiviral Vav3 (LV-VAV3) in MI rats upregulated Vav3 expression in MI heart tissue. Restoration of Vav3 expression reduced infarct area and ameliorated cardiac function in MI rats. Cardiac inflammation, apoptosis, and upregulation of NFκB signal in heart tissue of MI animals were assessed using ELISA, TUNEL staining, real-time PCR, and WB. Vav3 overexpression reduced cardiac inflammation and apoptosis and inhibited NFκB expression and activation. Betulinic Acid (BA) was then used to re-activate NFκB in Vav3-overexpressed and H2O2-induced cardiomyocytes. The expression of P50 and P65, as well as nuclear P65, was significantly increased by BA exposure. Conclusions Vav3 might serve as a target to reduce ischemia damage by suppressing the inflammation and apoptosis of cardiomyocytes. (MI failure Vav transducer activity However Vav3mediated Vavmediated mediated investigation Here artery H2O2 HO H O H2O2treated HOtreated treated LVVAV3 LVVAV LV VAV3 VAV (LV-VAV3 ELISA staining realtime real time PCR WB activation (BA reactivate re activate Vav3overexpressed Vavoverexpressed overexpressed H2O2induced HOinduced induced P P5 P65 exposure H2O (LV-VAV P6

Abstract Background Currently, only a few studies have described the general characteristics of patients with primary Sjögren's syndrome (pSS) who tested negatives for anti-SSA and anti-SSB antibodies. We aimed to further investigate the clinical characteristics of these patients in a large sample. Methods Data from patients with pSS who were treated at a tertiary hospital in China between 2013 and 2022 were retrospectively analyzed. Clinical characteristics of the patients were compared between those with and without anti-SSA and anti-SSB antibody negativity. Factors associated with anti-SSA and anti-SSB negativity were identified by logistic regression analysis. Results Overall, 934 patients with pSS were included in this study, among whom 299 (32.0%) tested negative for anti-SSA and anti-SSB antibodies. Compared with patients testing positive for anti-SSA or anti-SSB antibodies, that testing negative for the two antibodies had a lower proportion of females (75.3% vs. 90.6%, p < 0.001) and thrombocytopenia (6.7% vs. 13.6%, p = 0.002), but a higher proportion of abnormal Schirmer I tests (96.0% vs. 89.1%, p = 0.001) and interstitial lung disease (ILD) (59.2% vs. 28.8%, p = 0.001). Anti-SSA and anti-SSB negativity was positively associated with male sex (odds ratio [OR] = 1.86, 95% confidence interval [CI]: 1.05, 3.31), abnormal Schirmer I tests (OR = 2.85, 95% CI: 1.24, 6.53), and ILD (OR = 2.54, 95% CI: 1.67, 3.85). However, it was negatively related to thrombocytopenia (OR = 0.47, 95% CI: 0.24, 0.95). Conclusion Approximately one third of pSS patients had anti-SSA and anti-SSB negativity. pSS patients testing negative for anti-SSA and anti-SSB showed a higher risk of abnormal Schirmer I tests and ILD, but a lower risk of thrombocytopenia. Currently Sjögrens Sjögren s (pSS antiSSA anti SSA antiSSB SSB sample 201 202 analyzed analysis Overall 93 study 29 32.0% 320 32 0 (32.0% 75.3% 753 75 3 (75.3 vs 906 90 6 90.6% 0.001 0001 001 6.7% 67 7 (6.7 136 13 13.6% 0.002, 0002 0.002 , 002 0.002) 96.0% 960 96 (96.0 891 89 1 89.1% (ILD 59.2% 592 59 2 (59.2 288 28 8 28.8% 0.001. . AntiSSA Anti odds OR [OR 186 86 1.86 95 CI [CI] 105 05 1.05 3.31, 331 3.31 31 3.31) 285 85 2.85 124 24 1.24 6.53, 653 6.53 53 6.53) 254 54 2.54 167 1.67 3.85. 385 3.85 3.85) However 047 47 0.47 024 0.24 0.95. 095 0.95 0.95) 20 9 32.0 (32.0 75.3 (75. 90.6 0.00 000 00 6.7 (6. 13.6 96.0 (96. 89.1 59.2 5 (59. 28.8 18 1.8 [CI 10 1.0 33 3.3 2.8 12 1.2 65 6.5 25 2.5 16 1.6 38 3.8 04 4 0.4 02 0.2 09 0.9 32. (32. 75. (75 90. 0.0 6. (6 13. 96. (96 89. 59. (59 28. 1. 3. 2. 0. (32 (7 ( (9 (5 (3

Abstract Objectives: Lung cancer was one of the most common malignancies around the world. It has great significance in to search for the mechanism of occurrence and development of lung cancer. LIM Domain Binding protein 2 (LDB2) belongs to the LIM-domain binding family, it can be used as a binding protein that combined with other transcription factors to form the transcription complex for regulating the expression of target genes. The expression of microRNA-96-5p (miR-96-5p) has been investigated in various tumors. The aim of this study is to investigate the potential role of LDB2 and miR-96-5p in lung cancer. Methods: Real-time quantitative PCR was applied to detect the expression of LDB2 and miR-96-5p. The proliferation, invasion, and metastasis of H1299 cells were analyzed by CCK8, transwell, and wound healing assay after LDB2 or miR-96-5p transfection. Luciferase activities assay and western blot were used to reveal the targeted regulation between LDB2 and miR-96-5p. Results: Here the authors found LDB2 was down-regulated in lung cancer tissues and negatively correlated with miR-96-5p expression, it could promote or inhibit the proliferation, invasion and metastasis of H1299 cells after LDB2 knockdown or overexpression and regulate the expression of cyclinD1, MMP9, Bcl-2, and Bax via ERK1/2 signaling pathway. Furthermore, miR-96-5p exerted its function by directly binding to 3′-UTR of LDB2 and regulating expression of LDB2. miR-96-5p could promote the proliferation, invasion, and metastasis of H1299 cells. Conclusion: These findings demonstrate that LDB2 can act as a new regulator to inhibit cell proliferation, invasion, and metastasis via the ERK1/2 signaling pathway, and miR-96-5p may be a potential promising molecular by targeting LDB2 in lung cancer. Objectives world LDB (LDB2 LIMdomain domain family genes microRNA965p microRNAp microRNA 96 5p p miR965p miRp miR (miR-96-5p tumors Methods Realtime Real time miR965p. 5p. proliferation H H129 CCK8 CCK transwell transfection Results downregulated down regulated cyclinD1 cyclinD MMP9 MMP Bcl2, Bcl2 Bcl 2, Bcl-2 ERK12 ERK ERK1 ERK1/ pathway Furthermore 3′UTR 3UTR UTR 3′ 3 Conclusion (LDB 9 H12 Bcl- H1

Abstract Objective Immunotherapy has been proven to improve the prognosis of patients with advanced malignancy but has shown limited efficacy in patients with Colorectal Cancer (CRC). Increasing evidence suggests that butyrate, a bacterial metabolite, enhances the efficacy of cancer therapies by modulating immune responses. Here, the effect and the mechanism of butyrate on anti-PD-L1 therapy were investigated in CRC. Methods The expression of PD-L1 and STAT1, and the lysine acetylation of STAT1 in CRC cells were observed after treatment with butyrate (2, 5, and 10 mM) for 24h or butyrate (5 mM) for 8, 16, and 24h. Site-directed mutations of STAT1 (K410R or K413R) were introduced to determine the role of STAT1 acetylation in modulating PD-L1 expression. The effect of butyrate on the cytotoxicity of CD8+ T-cells against CRC cells with or without PD-L1 overexpression was explored in vitro and in vivo. Results Butyrate could suppress IFN-γ-induced PD-L1 up-regulation in CRC cells in a dose- and time-dependent way. Butyrate promoted the lysine acetylation of STAT1 to reduce STAT1 expression. Non-acetylated mutant STAT1 not only ameliorated butyrate-induced suppression of lysine acetylation and nuclear translocation of STAT1 but also blocked the effect of butyrate on PD-L1. Butyrate attenuated the IFN-γ-induced impairment of CD8+ T-cell cytotoxicity against CRC cells. Meanwhile, butyrate suppressed CRC tumor growth by enhancing CD8+ T-cell infiltration. However, directly overexpressing PD-L1 in CRC cells could abolish the effect of butyrate. Conclusion Butyrate strengthens the immune response to CRC cells by suppressing PD-L1 expression via acetylation of STAT1. . (CRC) metabolite responses Here antiPDL1 antiPDL anti PD L1 L anti-PD-L PDL1 PDL PD-L STAT 2, 2 (2 5 1 mM h ( 8 16 Sitedirected Site directed K410R KR K R K413R CD8 CD Tcells T vivo IFNγinduced IFN γ induced upregulation up regulation dose timedependent time dependent way Nonacetylated Non acetylated butyrateinduced PDL1. L1. Tcell cell Meanwhile infiltration However (CRC

ABSTRACT Salt-tolerant ‘Chuanqiao No.1’ and salt-sensitive ‘Chuanqiao No.2’ Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) were used as materials. Effects of ethephon on physiological characteristics of Tartary buckwheat under salt stress were studied by different concentrations of ethephon (ETH) under 150 mM NaCl stress. Ethephon treatment could improve seed germination rate of both varieties under salt stress, and the effect of ethephon treatment on salt-sensitive variety was better. Ethephon treatment could improve physiological characteristics of Tartary buckwheat seedlings under salt stress, and appropriate concentrations of ETH root treatment in salt-tolerant and salt-sensitive variety were 1.0 and 1.5 mM, and for ETH spraying were 0.4 and 0.6 mM. The effect of ETH spraying on improving physiological characteristics of Tartary buckwheat under salt stress was better than that of ETH root treatment. Salt tolerance gene FtNHX1 reached its maximum expression level at 24 h, which was increased by 277.77% and 251.17% in salt-tolerant and salt-sensitive variety compared with salt stress under ETH root treatment, while under ETH spraying treatment, which was increased by 232.39% and 190.91% in salt-tolerant and salt-sensitive variety compared with salt stress. Salt tolerance gene FtSOS1 reached its maximum expression levels at 12 h, which was increased by 60.94% and 98.75% in salt-tolerant and salt- sensitive variety compared with salt stress under ETH root treatment, while under ETH spraying treatment, which was increased by 35.98% and 67.63% in salt-tolerant and salt-sensitive variety compared with salt stress.

Abstract To observe effect and safety of different doses of ketorolac tromethamine in patient controlled intravenous analgesia (PCIA) after hip arthroplasty. 60 hip arthroplasty patients injected with 0.5 mg/kg ketorolac tromethamine before anesthesia were randomly divided into group A, B and C in which the patients were injected 2, 3 and 4 mg/kg ketorolac tromethamine after operation respectively. Compared with group A, MAP and HR in group B and group C were lower at T1 (3 h after operation) and T2 (6 h after operation). In group A, MAP and HR at T3 (24 h after operation) and T4 (48 h after operation) were lower than at T1. Compared with group A, NRS scores of group B and C were significantly lower during the analgesia. Compared with group A, ramesay sedation scores in group B and group C were significantly higher at T1 and T2; Harris hip scores of group B and group C were significantly higher at T3 and T4; the total number of pushing PCA pump and dezocine dosage were significantly decreased in group B and group C. Preemptive analgesia combined with postoperative analgesia on patients after hip replacement was appropriate.

Abstract Background: Pulmonary hypertension (PH) is a frequent complication of systemic sclerosis (SSc) and is currently one of the primary causes of death in patients with this disease. We conducted a systematic review and meta-analysis to assess the association between PH and mortality in patients with SSc to verify trends in mortality in patients with SSc-associated PH. Methods: We searched the PubMed and Embase databases for published studies on SSc-associated PH from inception to May 2021. All cohort studies in which mortality and/or survival for SSc-associated PH were reported were included in the analysis. The outcome parameters were pooled and analyzed using a random-effects model via generic inverse-variance weighting in conventional and cumulative meta-analysis. Results: The literature search identified 1161 citations, and the full texts of 54 studies were examined. Sixteen articles, with a total of 7857 patients with SSc and 1140 patients with SSc-associated PH, were included in the metaanalysis. Patients with SSc-associated PH had a higher pooled risk of mortality than patients with SSc without PH (risk ratio = 3.12; 95% confidence interval: [2.44, 3.98]). Conclusions: This meta-analysis revealed a higher mortality in patients with SSc-associated PH. PH was a significant predictor of death in patients with SSc. Thus, early diagnosis and treatment of PH are important in patients with SSc.

Abstract Genetically modified soybean strains are trade monitoring object by China. Currently, the genetically modified soybean MON89788 has been approved in China to be imported as processing raw material. Therefore, there is an urgent need to establish a method to quantitatively assess MON89788. This study used droplet digital PCR technology to quantitatively detect MON89788. The results showed that the genomic DNA concentration and its copy number of genetically modified soybeans showed a certain linear relationship. The formula was y = 2.5967x + 3.1437, where R2 = 0.999. When the same 4.5% genetically modified sample was contained, the results of the droplet digital PCR ratio and linear methods were 5.88% and 3.599%, respectively. Comparing the two results showed that the droplet digital PCR ratio method has a larger error than the linear relationship method. The novel droplet digital PCR linear relationship method established in this study has high sensitivity and specificity, and thus is a good prospect for quantitative research.

RESUMO Objetivo: Este estudo teve como objetivo avaliar o efeito da cobertura contínua do olho doente de pacientes com coriorretinopatia serosa central por 48 horas. Métodos: Este estudo retrospectivo, caso-controle, incluiu 32 pacientes com coriorretinopatia serosa central, dos quais 17 receberam tratamento de cobertura contínua por 48 horas no olho doente com gaze médica como grupo de tratamento e 15 foram acompanhados como grupo de observação. Todos os pacientes não receberam nenhum outro tratamento ou medicamento. O logaritmo do ângulo mínimo de resolução da acuidade visual melhor corrigida (LogMar), a espessura macular da retina e o valor médio da raiz quadrada da densidade da amplitude no primeiro anel do eletroretinograma multifocal foram examinados antes e após o tratamento por 48 horas, respectivamente. Resultados: Após o tratamento contínuo, a acuidade visual melhor corrigida pela escala logMar foi de 0, 31 ± 0, 18 no grupo de tratamento e 0, 56 ± 0, 37 no grupo de observação (p=0, 019). A espessura macular da retina foi de 461 ± 43 µm no grupo de tratamento e 498 ± 50 µm no grupo de observação (p=0,032). O valor médio da raiz quadrada da densidade de amplitude no primeiro anel do eletroretinograma multifocal foi de 32,5 ± 5,3 nV/deg2 no grupo com cobertura e foi de 26,6 ± 4,3 NV/deg2 no grupo de observação (p=0,002). Conclusões: O tratamento de cobertura contínua no olho doente, durante 48 horas, apresentou efeitos positivos na acuidade visual melhor corrigida, na espessura e na função macular da retina no tratamento da coriorretinopatia serosa central.

ABSTRACT Purpose: To assess the effect of continuously covering the sick eye affected with central serous chorioretinopathy for 48 h. Methods: This retrospective, case-control study involved 32 central serous chorioretinopathy patients categorized in the treatment group composed of 17 sick eye that received continuous covering treatment for 48 h with a medical gauze and the observation group composed of 15 of these patients who were followed up. None of the patients received any other treatments or medications. The logarithm of the minimal angle of resolution (logMAR) best-corrected visual acuity, macular retinal thickness, and the root mean square value of the amplitude density in the first ring of multifocal electroretinogram were examined before and after the 48-h treatment. Results: After the continuous treatment, the logMAR best-corrected visual acuities were 0.31 ± 0.18 and 0.56 ± 0.37 in the treatment and observation groups, respectively (p=0.019). The macular retinal thicknesses were 461 ± 43 µm and 498 ± 50 µm in the treatment and observation groups, respectively (p=0.032). The root mean square values of the amplitude density in the first ring of multifocal electroretinogram were 32.5 ± 5.3 nV/deg2 and 26.6 ± 4.3 nV/deg2 in the treatment and observation groups, respectively (p=0.002). Conclusions: The continuous application of the covering treatment for 48 h on the sick eye showed positive outcomes with respect to the best-corrected visual acuity, macular retinal thickness, and macular retina functions in the treatment of central serous chorioretinopathy.

Resumo Fundamento: A doença cardiovascular é a principal causa de morte em todo o mundo. A apoptose mediada por hipóxia em cardiomiócitos é uma das principais causas de distúrbios cardiovasculares. O tratamento com a proteína do fator de crescimento endotelial vascular (VEGF, do inglês vascular endothelial growth factor) foi testado, mas as dificuldades operacionais limitaram seu uso. Entretanto, com os avanços da terapia gênica, aumentou o interesse na terapia gênica baseada no VEGF em doenças cardiovasculares. No entanto, o mecanismo preciso pelo qual a reposição de VEGF resgata os danos pós-hipóxia em cardiomiócitos não é conhecido. Objetivos: Investigar o efeito da expressão de VEGF121 pós-hipóxia utilizando cardiomiócitos de ratos neonatos. Métodos: Cardiomiócitos isolados de ratos neonatos foram utilizados para estabelecer um modelo in vitro de lesão cardíaca induzida por hipóxia. O efeito da superexpressão de VEGF, isolado ou em conjunto com inibidores de moléculas pequenas que têm como alvo os canais de cálcio, receptores sensíveis ao cálcio (CaSR, do inglês calcium-sensitive receptors) e calpaína, no crescimento e proliferação celular em lesão de cardiomiócitos induzidos por hipóxia, foram determinados com ensaio de MTT, coloração TUNEL, coloração com Anexina V/PI, lactato desidrogenase e atividade da caspase. Para análise estatística, um valor de p<0,05 foi considerado significativo. Resultados: Verificou-se que o efeito do VEGF121 foi mediado por CaSR e calpaína, mas não foi dependente dos canais de cálcio. Conclusões: Nossos resultados, mesmo em um ambiente in vitro, estabelecem as bases para uma validação futura e testes pré-clínicos da terapia gênica baseada em VEGF em doenças cardiovasculares.

Abstract Background: Cardiovascular disease is the major cause of death worldwide. Hypoxia-mediated apoptosis in cardiomyocytes is a major cause of cardiovascular disorders. Treatment with vascular endothelial growth factor (VEGF) protein has been tested but operational difficulties have limited its use. However, with the advancements of gene therapy, interest has risen in VEGF-based gene therapy in cardiovascular disorders. However, the precise mechanism by which VEGF replenishment rescues post-hypoxia damage in cardiomyocytes is not known. Objectives: To investigate the effect of post-hypoxia VEGF121 expression using neonatal rat cardiomyocytes. Methods: Cardiomyocytes isolated from neonatal rats were used to establish an in vitro model of hypoxia-induced cardiac injury. The effect of VEGF overexpression, alone or in combination with small-molecule inhibitors targeting calcium channel, calcium sensitive receptors (CaSR), and calpain on cell growth and proliferation on hypoxia-induced cardiomyocyte injury were determined using an MTT assay, TUNEL staining, Annexin V/PI staining, lactate dehydrogenase and caspase activity. For statistical analysis, a value of P<0.05 was considered to be significant. Results: The effect of VEGF121 was found to be mediated by CaSR and calpain but was not dependent on calcium channels. Conclusions: Our findings, even though using an in vitro setting, lay the foundation for future validation and pre-clinical testing of VEGF-based gene therapy in cardiovascular diseases.

Fibrosis caused by the increase in extracellular matrix in cardiac fibroblasts plays an important role in the occurrence and development of atrial fibrillation (AF). The aim of this study was to investigate the role of hsa-miR-4443 in AF, human cardiac fibroblast (HCFB) proliferation, and extracellular matrix remodeling. TaqMan Stem-loop miRNA assay was used to measure hsa-miR-4443 expression in patients with persistent AF (n=123) and healthy controls (n=100). Patients with AF were confirmed to have atrial fibrosis by late gadolinium enhancement. At the cellular level, after hsa-miR-4443 mimic and inhibitor were transfected with HCFBs, proliferation, apoptosis, migration, and invasion were analyzed. Lastly, hsa-miR-4443-targeted gene and transforming growth factor (TGF)-β1/α-SMA/collagen pathway were evaluated by dual-luciferase reporter assay and western blot, respectively. In patients with AF, hsa-miR-4443 decreased significantly and collagen metabolism level increased significantly. Logistic regression analysis showed that low hsa-miR-4443 level was a risk factor of AF (P<0.001). The receiver operating characteristic curve revealed that hsa-miR-4443 was useful for predicting AF (area under the curve: 0.828, sensitivity: 0.71, specificity: 0.78, P<0.001). In HCFBs, hsa-miR-4443 targeted thrombospondin-1 (THBS1) and downregulated TGF-β1/α-SMA/collagen pathway. The inhibition of hsa-miR-4443 expression promoted HCFB proliferation, migration, invasion, myofibroblast differentiation, and collagen production. The significant reduction of hsa-miR-4443 can be used as a biomarker for AF. hsa-miR-4443 protected AF by targeting THBS1 and regulated TGF-β1/α-SMA/collagen pathway to inhibit HCFB proliferation and collagen synthesis.

Naringenin (NAR) is a major flavanone in citrus fruits that has multiple pharmacological attributes such as anticancer and antiatherogenic. This study aims to investigate the mechanism of NAR in high-fat-diet (HFD)-induced atherosclerosis (AS) in apolipoprotein E-knockout (ApoE-/-) mice. A HFD-induced AS ApoE-/- mouse model was established. The mice were treated with HFD, different doses of NAR and simvastatin (Simv). After drug treatment, the levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), superoxide dismutase (SOD), and alanine aminotransferase (ALT) were determined. The expression of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was detected using qRT-PCR and enzyme-linked immunosorbent assay. The plaque area of the aorta of AS mice was determined using oil red O staining. Western blot analysis was applied to measure the levels of autophagy-related proteins [protein 1 light chain 3B (LC3B), beclin 1, and p62]. The TC, TG, LDL-C, TNF-α, ALT, and MDA levels were significantly increased while the HDL-C, SOD, and GSH-Px levels were decreased in the HFD-induced AS ApoE-/- mice. NAR treatment reversed the expression of the above indicators in mice. After they were treated with different doses of NAR, the LC3B and beclin 1 levels were improved while the p62 protein level was decreased. This study suggested that NAR could promote cell autophagy to improve HFD-induced AS in ApoE-/- mice.

Resumo Introdução e objetivos Os mecanismos de neurotoxicidade dos anestésicos locais são complexos. A apoptose e a autofagia são mecanismos altamente organizados que mantêm a homeostase celular durante o estresse. Estudos revelam que a ativação da autofagia atua como mecanismo de proteção in vitro. Não está claro se a autofagia também desempenha essa função in vivo. O objetivo deste estudo foi analisar o papel da autofagia na neurotoxicidade induzida por anestésico local e esclarecer o mecanismo dessa neurotoxicidade utilizando um modelo de injeção intratecal em ratos. Métodos Dezoito ratos Sprague‐Dawley machos adultos saudáveis foram divididos aleatoriamente em três grupos. Antes de receber a injeção intratecal de bupivacaína a 1%, cada rato recebeu injeção intraperitoneal de veículo ou rapamicina (1 mg.kg‐1) uma vez ao dia durante 3 dias. As alterações patológicas foram examinadas por coloração com Hematoxilina e Eosina (HE). A apoptose foi analisada por coloração com o método dUTP Nick‐End Labeling (TUNEL) mediado por TdT. A expressão de caspase‐3, Beclin1 e LC3 foram examinadas por coloração Imunohistoquímica (IHQ). A expressão de Beclin1 e LC3 e a razão LC3‐II/LC3‐I foram detectadas por análise de western blot. Resultados Após a injeção intratecal de bupivacaína, ocorreu lesão patológica nos neurônios da medula espinhal e os níveis de apoptose e caspase‐3 aumentaram. A ativação da autofagia causada pela rapamicina mitigou de forma expressiva as alterações patológicas e diminuiu os níveis de apoptose e caspase‐3, aumentando a expressão de LC3 e Beclin1 e a razão LC3‐II/LC3‐I. Conclusões O aumento da autofagia diminui a apoptose dependente da caspase‐3 e melhora a sobrevivência neuronal in vivo. A ativação da autofagia pode ser uma estratégia terapêutica potencial para a neurotoxicidade induzida por anestésicos locais.

Abstract Background and objectives The mechanisms by which local anesthetics cause neurotoxicity are very complicated. Apoptosis and autophagy are highly coordinated mechanisms that maintain cellular homeostasis against stress. Studies have shown that autophagy activation serves as a protective mechanism in vitro. However, whether it also plays the same role in vivo is unclear. The aim of this study was to explore the role of autophagy in local anesthetic-induced neurotoxicity and to elucidate the mechanism of neurotoxicity in an intrathecally injected rat model. Methods Eighteen healthy adult male Sprague-Dawley rats were randomly divided into three groups. Before receiving an intrathecal injection of 1% bupivacaine, each rat received an intraperitoneal injection of vehicle or rapamycin (1 mg.kg-1) once a day for 3 days. The pathological changes were examined by Haematoxylin and Eosin (HE) staining. Apoptosis was analysed by TdT-mediated dUTP Nick-End Labelling (TUNEL) staining. Caspase-3, Beclin1 and LC3 expression was examined by Immunohistochemical (IHC) staining. Beclin1 and LC3 expression and the LC3-II/LC3-I ratio were detected by western blot analysis. Results After bupivacaine was injected intrathecally, pathological damage occurred in spinal cord neurons, and the levels of apoptosis and caspase-3 increased. Enhancement of autophagy with rapamycin markedly alleviated the pathological changes and decreased the levels of apoptosis and caspase-3 while increasing the expression of LC3 and Beclin1 and the ratio of LC3-II to LC3-I. Conclusions Enhancement of autophagy decreases caspase-3-dependent apoptosis and improves neuronal survivalin vivo. Activation of autophagy may be a potential therapeutic strategy for local anaesthetic-induced neurotoxicity.