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Porcine deltacoronavirus (PDCoV), an emerging enteropathogenic coronavirus, is a serious threat to piglets and has zoonotic potential. Here, we aimed to further explore the role of aminopeptidase N (APN) as a receptor for PDCoV and test the inhibitory effect of a chimeric APN protein strategy on PDCoV infection. PK-15 cells and LLC-PK1 cells expressing chimeric APN were selected and infected with PDCoV. Viral replication was significantly decreased in these chimeric APN cells compared with that in control group cells. To further characterize the effect of the chimeric APN strategy on PDCoV infection in vitro, primary intestinal epithelial cells isolated from chimeric APN pigs were inoculated with PDCoV. Viral challenge of these cells led to decreased PDCoV infection. More importantly, virally challenged chimeric APN neonatal piglets displayed reduced viral load, significantly fewer microscopic lesions in the intestinal tissue, and no diarrhea. Taken together, these findings deepen our understanding of the mechanism of PDCoV infection and provide a valuable model for the production of disease-resistant animals. IMPORTANCE: Porcine deltacoronavirus (PDCoV), an emerging enteropathogenic coronavirus, causes diarrhea in piglets and possesses the potential to infect humans. However, there are currently no effective measures for the prevention or control of PDCoV infection. Here, we have developed PK-15 cells, LLC-PK1 cells, and primary intestinal epithelial cells expressing chimeric APN, and viral challenge of these cells led to decreased PDCoV infection. Furthermore, virally challenged chimeric APN neonatal piglets displayed reduced viral load, significantly fewer microscopic lesions in the intestinal tissue, and no diarrhea. These data show that chimeric APN is a promising strategy to combat PDCoV infection.
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Animais Recém-Nascidos , Antígenos CD13 , Infecções por Coronavirus , Deltacoronavirus , Doenças dos Suínos , Replicação Viral , Animais , Suínos , Antígenos CD13/genética , Antígenos CD13/metabolismo , Doenças dos Suínos/virologia , Deltacoronavirus/genética , Infecções por Coronavirus/virologia , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/prevenção & controle , Carga Viral , Edição de Genes/métodos , Linhagem Celular , Células Epiteliais/virologia , Diarreia/virologiaRESUMO
BACKGROUND: Elevated plasma Lp-PLA2 (lipoprotein-associated phospholipase A2) activity is closely associated with an increased risk of cardiovascular events. However, whether and how Lp-PLA2 is directly involved in the pathogenesis of atherosclerosis is still unclear. To examine the hypothesis that Lp-PLA2 could be a potential preventative target of atherosclerosis, we generated Lp-PLA2 knockout rabbits and investigated the pathophysiological functions of Lp-PLA2. METHODS: Lp-PLA2 knockout rabbits were generated using CRISPR/Cas9 system to assess the role of Lp-PLA2 in plasma lipids regulation and identify its underlying molecular mechanisms. Homozygous knockout rabbits along with wild-type rabbits were fed a cholesterol-rich diet for up to 14 weeks and their atherosclerotic lesions were compared. Moreover, the effects of Lp-PLA2 deficiency on the key cellular behaviors in atherosclerosis were assessed in vitro. RESULTS: When rabbits were fed a standard diet, Lp-PLA2 deficiency led to a significant reduction in plasma lipids. The decreased protein levels of SREBP2 (sterol regulatory element-binding protein 2) and HMGCR (3-hydroxy-3-methylglutaryl coenzyme A reductase) in livers of homozygous knockout rabbits indicated that the cholesterol biosynthetic pathway was impaired with Lp-PLA2 deficiency. In vitro experiments further demonstrated that intracellular Lp-PLA2 efficiently enhanced SREBP2-related cholesterol biosynthesis signaling independently of INSIGs (insulin-induced genes). When fed a cholesterol-rich diet, homozygous knockout rabbits exhibited consistently lower level of hypercholesterolemia, and their aortic atherosclerosis lesions were significantly reduced by 60.2% compared with those of wild-type rabbits. The lesions of homozygous knockout rabbits were characterized by reduced macrophages and the expression of inflammatory cytokines. Macrophages of homozygous knockout rabbits were insensitive to M1 polarization and showed reduced DiI-labeled lipoprotein uptake capacity compared with wild-type macrophages. Lp-PLA2 deficiency also inhibited the adhesion between monocytes and endothelial cells. CONCLUSIONS: These results demonstrate that Lp-PLA2 plays a causal role in regulating blood lipid homeostasis and Lp-PLA2 deficiency protects against dietary cholesterol-induced atherosclerosis in rabbits. Lp-PLA2 could be a potential target for the prevention of atherosclerosis.
Assuntos
Aterosclerose , Hiperlipidemias , Animais , Coelhos , 1-Alquil-2-acetilglicerofosfocolina Esterase/genética , Lipoproteína(a) , Fosfolipases , Células Endoteliais/metabolismo , Aterosclerose/genética , Aterosclerose/prevenção & controle , Lipídeos , ColesterolRESUMO
BACKGROUND: Numerous previous research have established the need for spiritual care among patients with cancer globally. Nevertheless, there was limited research, primarily qualitative, on the spiritual care needs of Chinese inpatients with advanced breast cancer. Furthermore, the need for spiritual care was rarely explored using the Kano model. To better understand the spiritual care needs and attributes characteristics of inpatients with advanced breast cancer, this study examined the Kano model. METHODS: A descriptive cross-sectional design study was conducted in the oncology departments of three tertiary grade-A hospitals in China from October 2022 to May 2023. To guarantee high-quality reporting of the study, the Strengthening the Reporting of Observational Studies in Epidemiology Checklist was used. Data on the demographic characteristics questionnaire, the Nurse Spiritual Therapeutics Scale (NSTS), and the Kano model-based Nurse Spiritual Therapeutics Attributes Scale (K-NSTAs) were collected through convenience sampling. The Kano model, descriptive statistics, two independent samples t-tests, and one-way analysis of variance were used to analyze the data. RESULTS: The overall score for spiritual care needs was 31.16 ± 7.85. The two dimensions with the highest average scores, "create a good atmosphere" (3.16 ± 0.95), and the lowest average scores, "help religious practice" (1.72 ± 0.73). The 12 items were distributed as follows: three attractive attributes were located in Reserving Area IV; five one-dimensional attributes were distributed as follows: three one-dimensional attributes were located in Predominance Area I, and two were found in Improving Area II; two must-be attributes were located in Improving Area II; and two indifference attributes were located in Secondary Improving Area III. CONCLUSION: The Chinese inpatients with advanced breast cancer had a middle level of spiritual care needs, which need to be further improved. Spiritual care needs attributes were defined, sorted, categorized, and optimized accurately and perfectly by the Kano model. And "create a good atmosphere" and "share self-perception" were primarily one-dimensional and must-be attributes. In contrast, the items in the dimensions of "share self-perception" and "help thinking" were principally attractive attributes. Nursing administrators are advised to optimize attractive attributes and transform indifference attributes by consolidating must-be and one-dimensional attributes, which will enable them to take targeted spiritual care measures based on each patient's characteristics and unique personality traits.
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Neoplasias da Mama , Terapias Espirituais , Feminino , Humanos , Neoplasias da Mama/patologia , Neoplasias da Mama/psicologia , Neoplasias da Mama/terapia , China , Estudos Transversais , Pacientes Internados/psicologia , Espiritualidade , Inquéritos e QuestionáriosRESUMO
OBJECTIVES: To explore the postoperative kinesophobia of patients after percutaneous coronary intervention (PCI) and its related factors. BACKGROUND: Percutaneous coronary intervention is an effective method to treat coronary heart disease (CHD), and cardiac rehabilitation is an important auxiliary method after PCI. However, the compliance of patients with cardiac rehabilitation after PCI is not good, among which kinesophobia is an important influencing factor. DESIGN: A descriptive cross-sectional design was implemented, and the high-quality reporting of the study adhered to the Strengthening the Reporting of Observational Studies in Epidemiology Statement. METHODS: In total, 351 inpatients who underwent PCI in three tertiary grade-A hospitals in China were selected by convenient sampling method. We use one-way ANOVA and multiple linear regression analysis to determine the relevant related factors. RESULTS: The kinesophobia of patients after PCI was negatively correlated with chronic illness resource utilization and sense of personal mastery, and positively correlated with illness perception. Education level, clinical classification of CHD, exercise habits, chronic illness resource utilization, illness perception and sense of personal mastery entered the regression equation, which could explain 78.1% of the total variation. CONCLUSION: The level of kinesiophobia of patients after PCI is high. Education level, clinical classification of CHD, exercise habits, chronic illness resource utilization, illness perception and sense of personal mastery are the related factors of kinesiophobia of patients after PCI. RELEVANCE TO CLINICAL PRACTICE: By reducing the level of exercise fear of patients after PCI, patients are more likely to accept and adhere to the cardiac rehabilitation plan, thus improving their prognosis and improving their quality of life. PATIENT OR PUBLIC CONTRIBUTION: The patient underwent PCI in the research hospital. Researchers screen them according to the inclusion criteria and invite them to participate in this study. If they meet the requirements, participants will answer the research questionnaire face to face after signing the informed consent form.
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Hemophilia B is an X-linked recessive bleeding disorder caused by abnormalities in the coagulation factor IX gene. Without prophylactic treatment, patients experience frequent spontaneous bleeding episodes. Well-characterized animal models are valuable for determining the pathobiology of the disease and testing novel therapeutic innovations. Here, we generated a porcine model of hemophilia B using a combination of CRISPR/Cas9 and somatic cell nuclear transfer. Moreover, we tested the possibility of hemophilia B therapy by gene insertion. Frequent spontaneous joint bleeding episodes that occurred in hemophilia B pigs allowed a thorough investigation of the pathological process of hemophilic arthropathy. In contrast to the hemophilia B pigs, which showed a severe bleeding tendency and joint damage, the transgenic pigs carrying human coagulation factor IX exhibited a partial improvement of bleeding. In summary, this study not only offers a translational hemophilia B model for exploring the pathological process of hemophilic arthropathy but also provides a possibility for the permanent correction of hemophilia in the future by genome editing in situ.
Assuntos
Hemofilia A , Hemofilia B , Animais , Sistemas CRISPR-Cas , Fator IX/genética , Hemofilia A/genética , Hemofilia B/genética , Hemofilia B/terapia , Hemorragia/genética , Humanos , SuínosRESUMO
Cytidine base editors (CBEs) have been demonstrated to be useful for precisely inducing C:G-to-T:A base mutations in various organisms. In this study, we showed that the BE4-Gam system induced the targeted C-to-T base conversion in porcine blastocysts at an efficiency of 66.7-71.4% via the injection of a single sgRNA targeting a xeno-antigen-related gene and BE4-Gam mRNA. Furthermore, the efficiency of simultaneous three gene base conversion via the injection of three targeting sgRNAs and BE4-Gam mRNA into porcine parthenogenetic embryos was 18.1%. We also obtained beta-1,4-N-acetyl-galactosaminyl transferase 2, alpha-1,3-galactosyltransferase, and cytidine monophosphate-N-acetylneuraminic acid hydroxylase deficient pig by somatic cell nuclear transfer, which exhibited significantly decreased activity. In addition, a new CBE version (termed AncBE4max) was used to edit genes in blastocysts and porcine fibroblasts (PFFs) for the first time. While this new version demonstrated a three genes base-editing rate of 71.4% at the porcine GGTA1, B4galNT2, and CMAH loci, it increased the frequency of bystander edits, which ranged from 17.8 to 71.4%. In this study, we efficiently and precisely mutated bases in porcine blastocysts and PFFs using CBEs and successfully generated C-to-T and C-to-G mutations in pigs. These results suggest that CBEs provide a more simple and efficient method for improving economic traits, reducing the breeding cycle, and increasing disease tolerance in pigs, thus aiding in the development of human disease models.
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Citidina/genética , Edição de Genes/métodos , Suínos/genética , Animais , Blastocisto/metabolismo , Sistemas CRISPR-Cas , Galactosiltransferases/genética , Vetores Genéticos/genética , Oxigenases de Função Mista/genética , Mutagênese , N-Acetilgalactosaminiltransferases/genética , RNA Guia de Cinetoplastídeos/genética , Suínos/embriologiaRESUMO
Classical swine fever (CSF) caused by classical swine fever virus (CSFV) is one of the most detrimental diseases, and leads to significant economic losses in the swine industry. Despite efforts by many government authorities to stamp out the disease from national pig populations, the disease remains widespread. Here, antiviral small hairpin RNAs (shRNAs) were selected and then inserted at the porcine Rosa26 (pRosa26) locus via a CRISPR/Cas9-mediated knock-in strategy. Finally, anti-CSFV transgenic (TG) pigs were produced by somatic nuclear transfer (SCNT). Notably, in vitro and in vivo viral challenge assays further demonstrated that these TG pigs could effectively limit the replication of CSFV and reduce CSFV-associated clinical signs and mortality, and disease resistance could be stably transmitted to the F1-generation. Altogether, our work demonstrated that RNA interference (RNAi) technology combining CRISPR/Cas9 technology offered the possibility to produce TG animal with improved resistance to viral infection. The use of these TG pigs can reduce CSF-related economic losses and this antiviral strategy may be useful for future antiviral research.
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Antivirais , Peste Suína Clássica/prevenção & controle , Engenharia Genética/métodos , Animais , Animais Geneticamente Modificados , Vírus da Febre Suína Clássica , SuínosRESUMO
To investigate the therapeutic effects of cyclodextrin on the development of atherosclerosis in rabbits, we evaluated the effects of (2-hydroxypropyl)-ß-cyclodextrin (HPßCD) therapy on the organ coefficient, lipid profiles, inflammatory cytokines, and atherosclerotic plaques in rabbits fed a high-fat diet. Our results demonstrated that HPßCD therapy reduced plasma triglyceride levels and inflammatory cytokine levels but increased plasma high-density lipoprotein cholesterol levels. HPßCD therapy produced a significant decrease in the atherosclerotic lesion area and reduced macrophage and collagen content in the lesions. The expression levels of inflammatory genes in aortic plaques were significantly reduced by HPßCD treatment, but the expression of ATP-binding cassette (ABC) transporters A1 (ABCA1) and G1 (ABCG1) in aortic plaques and livers increased significantly. HPßCD therapy may produce additional antiatherosclerotic benefits likely via increasing high-density lipoprotein cholesterol levels.
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2-Hidroxipropil-beta-Ciclodextrina/farmacologia , Anti-Inflamatórios/farmacologia , Aorta/efeitos dos fármacos , Doenças da Aorta/prevenção & controle , Aterosclerose/prevenção & controle , HDL-Colesterol/sangue , Citocinas/sangue , Mediadores da Inflamação/sangue , Transportador 1 de Cassete de Ligação de ATP/metabolismo , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Aorta/metabolismo , Aorta/patologia , Doenças da Aorta/sangue , Doenças da Aorta/patologia , Aterosclerose/sangue , Aterosclerose/patologia , Biomarcadores/sangue , Dieta Hiperlipídica , Modelos Animais de Doenças , Progressão da Doença , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Placa Aterosclerótica , Coelhos , Regulação para CimaRESUMO
BACKGROUND: Arachidonic acid (AA) has potent pro-apoptotic effects on cancer cells at a low concentration and on macrophages at a very high concentration. However, the effects of AA on the macrophage cell cycle and related signaling pathways have not been fully investigated. Herein we aim to observe the effect of AA on macrophages cell cycle. RESULTS: AA exposure reduced the viability and number of macrophages in a dose- and time-dependent manner. The reduction in RAW264.7 cell viability was not caused by apoptosis, as indicated by caspase-3 and activated caspase-3 detection. Further research illustrated that AA exposure induced RAW264.7 cell cycle arrested at S phase, and some cell cycle-regulated proteins were altered accordingly. Moreover, JNK signaling was stimulated by AA, and the stimulation was partially reversed by a JNK signaling inhibitor in accordance with cell cycle-related factors. In addition, nuclear and total Foxo1/3a and phosphorylated Foxo1/3a were elevated by AA in a dose- and time-dependent manner, and this elevation was suppressed by the JNK signaling inhibitor. CONCLUSION: Our study demonstrated that AA inhibits macrophage viability by inducing S phase cell cycle arrest. The JNK signaling pathway and the downstream FoxO transcription factors are involved in AA-induced RAW264.7 cell cycle arrest.
Assuntos
Ácido Araquidônico/administração & dosagem , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Proteína Forkhead Box O1/genética , Macrófagos/efeitos dos fármacos , Animais , Caspase 3/genética , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , MAP Quinase Quinase 4/genética , Camundongos , Fosforilação , Células RAW 264.7 , Fase S/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
CRISPR/Cas9 has emerged as one of the most popular genome editing tools due to its simple design and high efficiency in multiple species. Myostatin (MSTN) negatively regulates skeletal muscle growth and mutations in myostatin cause double-muscled phenotype in various animals. Here, we generated myostatin mutation in Erhualian pigs using a combination of CRISPR/Cas9 and somatic cell nuclear transfer. The protein level of myostatin precursor decreased dramatically in mutant cloned piglets. Unlike myostatin knockout Landrace, which often encountered health issues and died shortly after birth, Erhualian pigs harboring homozygous mutations were viable. Moreover, myostatin knockout Erhualian pigs exhibited partial double-muscled phenotype such as prominent muscular protrusion, wider back and hip compared with wild-type piglets. Genome editing in Chinese indigenous pig breeds thus holds great promise not only for improving growth performance, but also for protecting endangered genetic resources.
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Animais Geneticamente Modificados/genética , Sistemas CRISPR-Cas , Mutação , Miostatina/genética , Suínos/genética , Animais , Feminino , Técnicas de Inativação de Genes , Homozigoto , Músculo Esquelético/fisiologia , Técnicas de Transferência Nuclear , Transfecção/métodosRESUMO
BACKGROUND: The consumption of n-3 polyunsaturated fatty acids (PUFAs) is important to human health, especially in cases of cardiovascular disease. Although beneficial effects of n-3 PUFAs have been observed in a number of studies, the mechanisms involved in these effects have yet to be discovered. METHODS: We generated hfat-1 transgenic pigs with traditional somatic cell nuclear transfer (SCNT) technology. The fatty acid composition in ear tissue of pigs were detected with gas chromatography. The cholesterol, triglycerides (TAG) and inflammation mediators in circulation were investigated. RESULTS: The hfat-1 transgenic pigs were developed which accumulate high levels of n-3 PUFAs than wild-types pigs. Gas chromatography results demonstrated that the total n-3 PUFAs in the ear tissues of the transgenic founders were 2-fold higher than the wild-type pigs. A lipid analysis demonstrated that the levels of TAG in the transgenic pigs were decreased significantly. The basal levels of the inflammation mediators tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in transgenic pigs were inhibited markedly compared with the wild-type pigs. CONCLUSIONS: These results suggest that n-3 PUFAs accumulation in vivo may have beneficial effects on vascular and hfat-1 transgenic pigs may be a useful tool for investigating the involved mechanisms.
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Animais Geneticamente Modificados , Caderinas/genética , Ácidos Graxos Ômega-3/farmacologia , Inflamação/dietoterapia , Triglicerídeos/sangue , Animais , Quimiocina CCL2/genética , Colesterol/sangue , Colesterol/genética , HDL-Colesterol/sangue , HDL-Colesterol/genética , Ácidos Graxos Ômega-3/farmacocinética , Feminino , Humanos , Inflamação/genética , Interleucina-6/genética , Masculino , Sus scrofa , Triglicerídeos/genética , Fator de Necrose Tumoral alfa/genéticaRESUMO
Lipoprotein-associated phospholipase A 2 (Lp-PLA2) is associated with the risk of vascular disease. It circulates in human blood predominantly in association with low-density lipoprotein cholesterol (LDL-C) and hydrolyses oxidized phospholipids into pro-inflammatory products. However, in the mouse circulation, it predominantly binds to high-density lipoprotein cholesterol (HDL-C) and exhibits anti-inflammatory properties. To further investigate the effects of Lp-PLA2 in the circulation, we generated over-expressed Lp-PLA2 transgenic swine. The eukaryotic expression plasmid of porcine Lp-PLA2 which driven by EF1α promoter was constructed and generate transgenic swine via SCNT. The expression and activity of Lp-PLA2 in transgenic swine were evaluated, and the total cholesterol (TC), HDL-C, LDL-C and triglyceride (TG) levels in the fasting and fed states were also assessed. Compared with wild-type swine controls, the transgenic swine exhibited elevated Lp-PLA2 mRNA levels and activities, and the activity did not depend on the feeding state. The TC, HDL-C and LDL-C levels were not significantly increased. There was no change in the TG levels in the fasting state between transgenic and control pigs. However, in the fed state, the TG levels of transgenic swine were slightly increased compared with the control pigs and were significantly elevated compared with the fasting state. In addition, inflammatory gene (interleukin [IL]-6, monocyte chemotactic protein [MCP]-1 and tumor necrosis factor [TNF]-α) mRNA levels in peripheral blood mononuclear cells (PBMCs) were significantly increased. The results demonstrated that Lp-PLA2 is associated with triglycerides which may be helpful for understanding the relationship of this protein with cardiovascular disease.
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1-Alquil-2-acetilglicerofosfocolina Esterase/genética , 1-Alquil-2-acetilglicerofosfocolina Esterase/imunologia , Citocinas/imunologia , Leucócitos Mononucleares/imunologia , Suínos/genética , Suínos/imunologia , Animais , Animais Geneticamente Modificados/genética , Animais Geneticamente Modificados/imunologia , Células Cultivadas , Regulação para Cima/genéticaRESUMO
OBJECTIVE: To observe the effect of Chuanhuang No.1 Recipe (CHR) on renal function and micro-inflammation in phase 3 chronic kidney disease (CKD) patients. METHODS: Totally 60 phase 3 CKD patients were randomly assigned to the treatment group (treated by CHR) and the control group (treated by Losartan Potassium), 30 in each group. All patients received basic treatment. Patients in the treatment group took CHR decoction, 400 mL each time, one dose per day, while those in the control group took Losartan Potassium, 50-100 mg per day. All medication lasted for 24 weeks. Changes of serum creatinine (SCr), blood urea nitrogen (BUN), estimated glomerular filtration rate (eGFR), serum uric acid (UA), 24 h urinary protein excretion (24 h U-pro), urinary microalbumin (U-Alb), high-sensitivity C-reactive protein (hs-CRP), serum tumor necrosis factor (TNF)-alpha, and serum IL-6 were detected and compared before and after treatment. Efficacy was also compared. RESULTS: Compared with before treatment, SCr and BUN significantly decreased in the treatment group (P<0.05, P<0.01); eGFR in- creased (P<0.05). Only UA obviously decreased in the control group (P<0.05), but with no obvious change in SCr, BUN, or eGFR. Compared with before treatment, 24 h U-pro decreased after treatment in the treatment group (P<0.05), but with less decreased level when compared with the control group. U- Alb was also significantly decreased in the control group (P<0.01). There was statistical difference in 24 h U-pro and U-Alb between the two groups after treatment (P<0.05). Compared with before treatment, hs-CRP obviously decreased after treatment in the two groups, but serum levels of TNF-alpha and IL-6 obviously decreased only in the treatment group (P<0.05). The total effective rate was obviously higher in the treatment group than in the control group (70.00% vs. 43.33%, P<0.01). CONCLUSION: CHR could efficiently improve the renal function of phase 3 CKD patients and alleviate the micro-inflammation.
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Medicamentos de Ervas Chinesas/uso terapêutico , Insuficiência Renal Crônica/tratamento farmacológico , Adulto , Nitrogênio da Ureia Sanguínea , Proteína C-Reativa/metabolismo , Feminino , Humanos , Inflamação , Interleucina-6/metabolismo , Losartan/uso terapêutico , Masculino , Pessoa de Meia-Idade , Fitoterapia , Fator de Necrose Tumoral alfa/metabolismo , UreiaRESUMO
The developmental ability among embryos produced by three different techniques were examined: there were no significant differences in the developmental rate in porcine embryos produced by in vitro fertilization (IVF) and first generation of somatic cell nucleus transfer (SCNT), but the developmental rate dropped sharply at the 2- to four-cell stage in recloned (second generation of SCNT) embryos. In most recloned embryos, Oct4 and Klf4 were under-expressed at all stages, whereas Sox2 and Nanog were over-expressed at the two-cell stage. In contrast, Nanog was absent in IVF and SCNT embryos at the two-cell stage. The recloned embryos were treated with valproic acid to enhance developmental capacity and this led to an increase in the rate of blastocyst formation and total cell number compared with the findings for untreated recloned embryos (29.8 vs. 12.4 %, 39 vs. 25, respectively, p < 0.05).
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Perfilação da Expressão Gênica/métodos , Suínos/embriologia , Animais , Fertilização in vitro , Técnicas de Transferência Nuclear , Ácido Valproico/metabolismoRESUMO
Pseudorabies virus (PRV), an alphaherpesvirus, is a neglected zoonotic pathogen. Dectin-1 sensing of ß-glucan (BG) induces trained immunity, which can possibly form a new strategy for the prevention of viral infection. However, alphaherpesvirus including PRV have received little to no investigation in the context of trained immunity. Here, we found that BG pretreatment improved the survival rate, weight loss outcomes, alleviated histological injury and decreased PRV copy number of tissues in PRV-infected mice. Type I interferons (IFNs) including IFN-α/ß levels in serum were significantly increased by BG. However, these effects were abrogated in the presence of Dectin-1 antagonist. Dectin-1-mediated effect of BG was also confirmed in porcine and murine macrophages. These results suggested that BG have effects on type I IFNs with antiviral property involved in Dectin-1. In piglets, oral or injected immunization with BG and PRV vaccine could significantly elevated the level of PRV-specific IgG and type I IFNs. And it also increased the antibody levels of porcine reproductive and respiratory syndrome virus vaccine and classical swine fever vaccine that were later immunized, indicating a broad-spectrum effect on improving vaccine immunity. On the premise that the cost was greatly reducing, the immunological effect of oral was better than injection administration. Our findings highlighted that BG induced type I IFNs related antiviral effect against PRV involved in Dectin-1 and potential application value as a feed additive to help control the spread of PRV and future emerging viruses.
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Herpesvirus Suídeo 1 , Interferon Tipo I , Lectinas Tipo C , Pseudorraiva , beta-Glucanas , Animais , beta-Glucanas/farmacologia , beta-Glucanas/administração & dosagem , Camundongos , Suínos , Lectinas Tipo C/imunologia , Pseudorraiva/imunologia , Pseudorraiva/prevenção & controle , Interferon Tipo I/imunologia , Herpesvirus Suídeo 1/imunologia , Herpesvirus Suídeo 1/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/efeitos dos fármacos , Antivirais/farmacologia , Vacinas Virais/imunologia , FemininoRESUMO
Charcot-Marie-Tooth type 2A (CMT2A) is a single-gene motor sensory neuropathy caused by Mfn2 mutation. It is generally believed that CMT2A involves mitochondrial fusion disruption. However, how Mfn2 mutation mediates the mitochondrial membrane fusion loss and its further pathogenic mechanisms remain unclear. Here, in vivo and in vitro mouse models harboring the Mfn2R364W, Mfn2G176S and Mfn2H165R mutations were constructed. Mitochondrial membrane fusion and fission proteins analysis showed that Mfn2R364W, Mfn2G176S, and Mfn2H165R/+ mutations maintain the expression of Mfn2, but promote Drp1 upregulation and Opa1 hydrolytic cleavage. In Mfn2H165R/H165R mutation, Mfn2, Drp1, and Opa1 all play a role in inducing mitochondrial fragmentation, and the mitochondrial aggregation is affected by Mfn2 loss. Further research into the pathogenesis of CMT2A showed these three mutations all induce mitochondria-mediated apoptosis, and mitochondrial oxidative phosphorylation damage. Overall, loss of overall fusion activity affects mitochondrial DNA (mtDNA) stability and causes mitochondrial loss and dysfunction, ultimately leading to CMT2A disease. Interestingly, the differences in the pathogenesis of CMT2A between Mfn2R364W, Mfn2G176S, Mfn2H165R/+ and Mfn2H165R/H165R mutations, including the distribution of Mfn2 and mitochondria, the expression of mitochondrial outer membrane-associated proteins (Bax, VDAC1 and AIF), and the enzyme activity of mitochondrial complex I, are related to the expression of Mfn2.
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BACKGROUND: The significance of problem-solving ability has been confirmed in numerous studies worldwide, highlighting its role in enhancing the skills of nursing interns and reducing psychological pressure. However, existing research indicates that the problem-solving ability of nursing interns urgently needs to be further improved. Limited research has been conducted on the problem-solving ability of nursing interns, and the correlations among problem-solving ability, future time perspective, and future work self of Chinese nursing interns are unclear. OBJECTIVES: To investigate problem-solving ability, future time perspective, and future work self among the Chinese nursing interns, and to examine the relationships among these variables. Additionally, the study aims to explore the mediating role of future work self between problem-solving ability and future time perspective. METHODS: A cross-sectional and correlational design was employed, adhering to the quality reporting conformed to the STROBE Checklist. From May 8, 2023, to February 15, 2024, 1,251 nursing interns were recruited from 15 tertiary grade-A hospitals across six cities in China. The Demographic Characteristics Questionnaire, Social Problem-Solving Inventory, Future Time Perspective Inventory, and Future Work Self Scale were used. The data was analyzed using descriptive statistics, univariate, correlation, and process plug-in mediation effect analyses. RESULTS: The total scores for problem-solving ability, future time perspective, and future work self were 64.39 ± 18.55, 45.08 ± 11.37, and 16.92 ± 5.28, respectively. Problem-solving ability was positively correlated with future time perspective (r = 0.638, p < 0.001) and future work self (r = 0.625, p < 0.001). Additionally, future work self partially mediated mediating role between problem-solving ability and future time perspective, accounting for 39.7% of the total effect. CONCLUSION: The problem-solving ability, future time perspective, and future work self among the Chinese nursing interns were relatively moderate, indicating a need for improvement. It is suggested that nursing managers and educators should actively implement career management and planning programs. By enhancing the future time perspective and future work self of nursing interns, their problem-solving ability can be improved. This, in turn, will facilitate their adaptation to clinical work, enhance the quality of nursing care, and promote the development of their nursing profession.
Assuntos
Resolução de Problemas , Humanos , Feminino , Masculino , China , Estudos Transversais , Adulto , Inquéritos e Questionários , Internato e Residência , Estudantes de Enfermagem/psicologia , Adulto Jovem , População do Leste AsiáticoRESUMO
BACKGROUND: During the Omicron pandemic, clinical first-line nurses played a crucial role in healthcare. Their innovative behavior enhanced the quality of nursing and served as a vital factor in driving the sustainable development of the nursing discipline and healthcare industry. Many previous studies have confirmed the significance of nurses' innovative behavior worldwide. However, the correlations among innovative behaviors, organizational innovation climate, self-transcendence, and their mediating roles in Chinese clinical first-line nurses need further research. METHODS: A cross-sectional study was conducted, and the quality reporting conformed to the STROBE Checklist. From March 2022 to February 2023, a convenience sample of 1,058 Chinese clinical first-line nurses was recruited from seven tertiary grade-A hospitals of Tianjin city in Northern China. The Demographic Characteristics Questionnaire, Nurse Innovative Behavior Scale (NIBS), Nurse Organizational Innovation Climate Scale, and the Self-Transcendence Scale were used. The data was analyzed using descriptive statistics, correlation, and process plug-in mediation effect analyses. RESULTS: The total scores of innovative behavior, organizational innovation climate, and self-transcendence were 33.19 ± 6.71, 68.88 ± 12.76, and 41.25 ± 7.83, respectively. Innovative behavior was positively correlated with the organizational innovation climate (r = 0.583, p < 0.01) and self-transcendence (r = 0.635, p < 0.01). Self-transcendence partially mediated mediating role between innovative behavior and organizational innovation climate, accounting for 41.7%. CONCLUSION: The innovative behavior, organizational innovation climate, and self-transcendence among the first-line nurses during the Omicron pandemic were relatively moderate, which needs improving. Organizational innovation climate can directly affect the innovative behavior among Chinese clinical first-line nurses and indirectly through the mediating role of self-transcendence. It is recommended that nursing managers adjust their management strategies and techniques based on the unique characteristics of nurses during the pandemic. This includes fostering a positive and inclusive environment for organizational innovation, nurturing nurses' motivation and awareness for innovation, enhancing their ability to gather information effectively, overcoming negative emotions resulting from the pandemic, and promoting personal growth. These efforts will ultimately enhance nursing quality and satisfaction during the Omicron pandemic.
Assuntos
COVID-19 , Inovação Organizacional , Humanos , COVID-19/epidemiologia , China/epidemiologia , Estudos Transversais , Adulto , Feminino , Masculino , Inquéritos e Questionários , Pandemias , Enfermeiras e Enfermeiros/psicologia , SARS-CoV-2 , Cultura Organizacional , Pessoa de Meia-Idade , População do Leste AsiáticoRESUMO
Porcine reproductive and respiratory syndrome (PRRS) is a globally prevalent contagious disease caused by the positive-strand RNA PRRS virus (PRRSV), resulting in substantial economic losses in the swine industry. Modifying the CD163 SRCR5 domain, either through deletion or substitution, can eff1ectively confer resistance to PRRSV infection in pigs. However, large fragment modifications in pigs inevitably raise concerns about potential adverse effects on growth performance. Reducing the impact of genetic modifications on normal physiological functions is a promising direction for developing PRRSV-resistant pigs. In the current study, we identified a specific functional amino acid in CD163 that influences PRRSV proliferation. Viral infection experiments conducted on Marc145 and PK-15 CD163 cells illustrated that the mE535G or corresponding pE529G mutations markedly inhibited highly pathogenic PRRSV (HP-PRRSV) proliferation by preventing viral binding and entry. Furthermore, individual viral challenge tests revealed that pigs with the E529G mutation had viral loads two orders of magnitude lower than wild-type (WT) pigs, confirming effective resistance to HP-PRRSV. Examination of the physiological indicators and scavenger function of CD163 verified no significant differences between the WT and E529G pigs. These findings suggest that E529G pigs can be used for breeding PRRSV-resistant pigs, providing novel insights into controlling future PRRSV outbreaks.
Assuntos
Antígenos CD , Antígenos de Diferenciação Mielomonocítica , Mutação Puntual , Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Receptores de Superfície Celular , Animais , Suínos , Síndrome Respiratória e Reprodutiva Suína/genética , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Antígenos de Diferenciação Mielomonocítica/genética , Antígenos de Diferenciação Mielomonocítica/metabolismo , Antígenos CD/genética , Antígenos CD/metabolismo , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Animais Geneticamente Modificados/genética , Linhagem CelularRESUMO
BACKGROUND: Apoptosis is recognized as an important mechanism in contrast-induced nephropathy (CIN). As tetramethylpyrazine (TMP) has been recently found to be renoprotective and anti-apoptotic in multiple kidney injuries, we hypothesized that TMP would prevent CIN. METHODS: An experimental model of CIN was established in rats. Serum creatinine, blood urea nitrogen, plasma cystatin C, urinary N-acetyl-ß-glucosaminidase, and urinary γ-glutamyl transpeptidase were measured to evaluate kidney function. Apoptosis was assessed by transmission electron microscopy, transferase-mediated deoxyuridine triphosphate nick end-labeling staining, and poly-ADP-ribose polymerase cleavage. Fork-head box O1 transcriptional factor (FoxO1) mRNA expression was evaluated by quantitative real-time PCR. Phospho-p38 mitogen-activated protein kinase (MAPK) protein expression was assessed by immunohistochemistry and Western blotting. RESULTS: TMP significantly attenuated the resulting renal dysfunction and renal tubular cell apo-ptosis. Mechanistically, TMP decreased the expression of phospho-p38 MAPK protein and attenuated the increased FoxO1 mRNA and nuclear protein expression. In addition, TMP inhibited inducible nitric oxide synthase and Bax protein expression while it upregulated Bcl-2. CONCLUSION: In summary, this study demonstrated the protective role of TMP against CIN and indicated the effects of TMP may be mediated by the inhibition of p38 MAPK and FoxO1 pathways. Thus, TMP may be a new potential therapeutic agent to prevent CIN.