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The primary hurdles for small interference RNA (siRNA) in clinical use are targeted and cytosolic delivery. To overcome both challenges, we have established a novel platform based on phage display, called NNJA. In this approach, a lysosomal cathepsin substrate is engineered within the flexible loops of PIII, that is displaying a unique random sequence at its N-terminus. NNJA library selection targeting cell-expressed targets should yield specific peptides localized in the cytoplasm. That is because phage internalization and subsequent localization to lysosome, upon peptide binding to the cell expressed target, will result in cleavage of PIII, rendering phage non-infective. Such phage will be eliminated from the selected pool and only peptide-phage that escapes lysosomes will advance to the next round. Proof of concept studies with the NNJA library demonstrated cytosolic localization of selected peptide-phage and peptide-siRNA, confirmed through confocal microscopy. More importantly, conjugation of siHPRT to monomeric or multimeric NNJA peptides resulted in significant reduction in HPRT mRNA in various cell types without significant cytotoxicity. Sequence similarity and clustering analysis from NGS dataset provide insights into sequence composition facilitating cell penetration. NNJA platform offers a highly efficient peptide discovery engine for targeted delivery of oligonucleotides to cytosol.
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Péptidos de Penetración Celular , Biblioteca de Péptidos , ARN Interferente Pequeño , Péptidos de Penetración Celular/metabolismo , Péptidos de Penetración Celular/química , Humanos , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Lisosomas/metabolismo , Técnicas de Visualización de Superficie Celular/métodos , Citosol/metabolismoRESUMEN
BACKGROUND: Cervical cancer (CC) remains a significant clinical challenge, even though its fatality rate has been declining in recent years. Particularly in developing countries, the prognosis for CC patients continues to be suboptimal despite numerous therapeutic advances. METHODS: Using The Cancer Genome Atlas database, we extracted CC-related data. From this, 52 methylation-related genes (MRGs) were identified, leading to the selection of a 10 long non-coding RNA (lncRNA) signature co-expressed with these MRGs. R programming was employed to filter out the methylation-associated lncRNAs. Through univariate, least absolute shrinkage and selection operator (i.e. LASSO) and multivariate Cox regression analysis, an MRG-associated lncRNA model was constructed. The established risk model was further assessed via the Kaplan-Meier method, principal component analysis, functional enrichment annotation and a nomogram. Furthermore, we explored the potential of this model with respect to guiding immune therapeutic interventions and predicting drug sensitivities. RESULTS: The derived 10-lncRNA signature, linked with MRGs, emerged as an independent prognostic factor. Segmenting patients based on their immunotherapy responses allowed for enhanced differentiation between patient subsets. Lastly, we highlighted potential compounds for distinguishing CC subtypes. CONCLUSIONS: The risk model, associated with MRG-linked lncRNA, holds promise in forecasting clinical outcomes and gauging the efficacy of immunotherapies for CC patients.
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Adenina/análogos & derivados , ARN Largo no Codificante , Neoplasias del Cuello Uterino , Humanos , Femenino , Pronóstico , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/terapia , ARN Largo no Codificante/genética , InmunoterapiaRESUMEN
Clear cell renal cell carcinoma (ccRCC) stands as a challenging subtype of kidney cancer, frequently complicating patient prognosis due to factors like postsurgical recurrences or late-stage diagnoses. In this study, we employed bioinformatics to investigate lactylation modifications in ccRCC, focusing on the TCGA-KIRC cohort. Out of 328 lactylation-associated genes, 31 emerged as differentially expressed, with 16 showing a marked correlation with overall survival. These genes exhibited strong protein-protein interactions and significant expression correlations. Intriguingly, a notable loss of gene copy numbers suggests potential implications in tumor progression. Utilizing unsupervised clustering, KIRC samples were grouped into two distinct subcategories, each showcasing different survival outcomes. While pathway enrichment highlighted an aggressive, inflammation-driven profile for subgroup 2, subgroup 1 was characterized by metabolic prominence. Furthermore, subgroup 2 presented an intensified inflammatory response, hinting at potential immune exhaustion. Capitalizing on machine learning, we developed a risk model using the TCGA-KIRC dataset, efficiently categorizing ccRCC patients into high- and low-risk clusters. Notably, those in the low-risk group indicated a more favorable survival trajectory. Clinical evaluations further corroborated these findings, linking better outcomes with reduced risk scores. Additionally, observed mutation patterns allude to a potential association between elevated risk scores and cytokine storms. TIDE analysis illuminated possible immunotherapeutic benefits for the low-risk group, underscored by an evident rise in microsatellite instability. Finally, our drug sensitivity evaluations revealed distinct therapeutic responses between the groups. In summary, this research underscores the pivotal role of lactylation modifications in ccRCC and introduces a promising prognostic model. These revelations pave the way for enhanced prognostic precision, presenting a promising path toward personalized treatment strategies and enriching our comprehension of the multifaceted molecular landscape of the disease.
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Carcinoma de Células Renales , Carcinoma , Neoplasias Renales , Humanos , Pronóstico , Factores de RiesgoRESUMEN
Abnormal or excessive accumulation of adipose tissue leads to a condition called obesity. Long-term positive energy balance arises when energy intake surpasses energy expenditure, which increases the risk of metabolic and other chronic diseases, such as atherosclerosis. In industrialized countries, the prevalence of coronary heart disease is positively correlated with the human development index. Atherosclerotic cardiovascular disease (ACD) is among the primary causes of death on a global scale. There is evidence to support the notion that individuals from varied socioeconomic origins may experience varying mortality effects as a result of high blood pressure, high blood sugar, raised cholesterol levels, and high body mass index (BMI). However, it is believed that changes in the concentration of trace elements in the human body are the main contributors to the development of some diseases and the transition from a healthy to a diseased state. Metal trace elements, non-metal trace elements, and the sampling site will be examined to determine whether trace elements can aid in the diagnosis of atherosclerosis. This article will discuss whether trace elements, discussed under three sections of metal trace elements, non-metal trace elements, and the sampling site, can participate in the diagnosis of atherosclerosis.
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Aim: Coronary heart disease is a major cause of mortality in developed and developing countries. Changes in the trace element concentration in the human body are one of the main reasons for the transition of the human body from a healthy to a diseased state. In this meta-analysis, we have studied the relationship between the reduction in serum zinc ion concentration and coronary heart disease. Methods: We used PubMed and Cochrane (as of June 30, 2021) databases for the literature search. Per the requirements of this systematic review, case-control studies involving serum zinc ion concentration and coronary heart disease were searched, and the quality of the included studies was evaluated before the meta-analysis. Results: A total of 3,981 cases were found across seven articles. The standard mean deviation (SMD) of serum zinc ion concentration was -0.22 [-0.28, -0.15], z = 6.52, and P < 0.05 indicated that the difference was statistically significant. The forest plot results show that I 2 = 34% < 50%, and the Q test showed P=0.17 > 0.1. These results suggest a lack of heterogeneity among the selected articles. Results from the funnel chart indicated that this study was free from publication bias. Conclusion: The results of this meta-analysis reveal that a decrease in serum zinc ion concentration is related to the occurrence of coronary heart disease. Clinically, monitoring the serum zinc ion levels is proven to be of great significance for patients with coronary heart disease.
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Coronary atherosclerosis is a potentially chronic circulatory condition that endangers human health. The biological cause underpinning cardiovascular disease is coronary atherosclerosis, and acute cardiovascular events can develop due to thrombosis, platelet aggregation, and unstable atherosclerotic plaque rupture. Coronary atherosclerosis is progressive, and three specific changes appear, with fat spots and stripes, atherosclerosis and thin-walled fiber atherosclerosis, and then complex changes in arteries. The progression and severity of cardiovascular disease are correlated with various levels of calcium accumulation in the coronary artery. The therapy and diagnosis of coronary atherosclerosis benefit from the initial assessment of the size and degree of calcification. This article will discuss the new progress in the early diagnosis of coronary atherosclerosis in terms of three aspects: imaging, gene and protein markers, and trace elements. This study intends to present the latest methods for diagnosing patients with early atherosclerosis through a literature review.
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Aterosclerosis , Enfermedades Cardiovasculares , Enfermedad de la Arteria Coronaria , Placa Aterosclerótica , Aterosclerosis/diagnóstico , Angiografía Coronaria , Enfermedad de la Arteria Coronaria/diagnóstico , Vasos Coronarios , Diagnóstico Precoz , Humanos , Placa Aterosclerótica/diagnóstico por imagen , Factores de RiesgoRESUMEN
Objective: Our study's goal was to find out acute myocardial infarction (AMI) patients' NUMB gene expression patterns and to evaluate its role as a diagnostic marker for AMI detection. Methods: Peripheral blood was drawn from 124 individuals who had an AMI and 115 patients who had stable coronary artery disease (SCAD). The real-time quantitative polymerase chain reaction was used to measure the mRNA expression level of the NUMB gene in peripheral blood. Results: The AMI group's NUMB gene expression was 0.906 (0.181-0.954), whereas the SCAD group's expression was 1.024 (0.207-1.127). However, the AMI group had 0.885 times lower NUMB mRNA expression than the SCAD group (P < 0.05). Conclusion: Multivariate logistic regression evaluation found that lower NUMB expression was correlated with an increased risk of coronary artery disease. However, age and fasting plasma glucose levels were not associated with decreased NUMB expression.
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Objective: To explore the cardioprotective effects of exercise-derived ß-aminoisobutyric (BAIBA) on cardiomyocyte apoptosis and energy metabolism in a rat model of heart failure (HF). Methods: In male Sprague-Dawley rats (8-week-old), myocardial infarction (MI) was used to induce HF by ligating the left anterior descending branch of the coronary artery. In the Sham group, the coronary artery was threaded but not ligated. After HF development, Sham and HF rats were exercised 60 min daily, 5 days/week on a treadmill for 8 weeks (50-60% maximal intensity) and exercise-induced cardiac remodeling after MI were assessed using echocardiography, hematoxylin and eosin (H&E), Masson's Trichrome, and TUNEL staining for the detection of apoptosis-associated factors in cardiac tissue. High-throughput sequencing and mass spectrometry were used to measure BAIBA production and to explore its cardioprotective effects and molecular actions. To further characterize the cardioprotective effects of BAIBA, an in vitro model of apoptosis was generated by applying H2 O 2 to H9C2 cells to induce mitochondrial dysfunction. In addition, cells were transfected with either a miR-208b analog or a miR-208b inhibitor. Apoptosis-related proteins were detected by Western Blotting (WB). ATP production was also assessed by luminometry. After administration of BAIBA and Compound C, the expression of proteins related to apoptosis, mitochondrial function, lipid uptake, and ß-oxidative were determined. Changes in the levels of reactive oxygen species (ROS) were assessed by fluorescence microscopy. In addition, alterations in membrane potential (δψm) were obtained by confocal microscopy. Results: Rats with HF after MI are accompanied by mitochondrial dysfunction, metabolic stress and apoptosis. Reduced expression of apoptosis-related proteins was observed, together with increased ATP production and reduced mitochondrial dysfunction in the exercised compared with the Sham (non-exercised) HF group. Importantly, exercise increased the production of BAIBA, irrespective of the presence of HF. To assess whether BAIBA had similar effects to exercise in ameliorating HF-induced adverse cardiac remodeling, rats were treated with 75 mg/kg/ day of BAIBA and we found BAIBA had a similar cardioprotective effect. Transcriptomic analyses found that the expression of miR-208b was increased after BAIBA administration, and subsequent transfection with an miR-208b analog ameliorated both the expression of apoptosis-related proteins and energy metabolism in H2O2-treated H9C2 cells. In combining transcriptomic with metabolomic analyses, we identified AMPK as a downstream target for BAIBA in attenuating metabolic stress in HF. Further cell experiments confirmed that BAIBA increased AMPK phosphorylation and had a cardioprotective effect on downstream fatty acid uptake, oxidative efficiency, and mitochondrial function, which was prevented by the AMPK inhibitor Compound C. Conclusion: Exercise-generated BAIBA can reduce cardiomyocyte metabolic stress and apoptosis induced by mitochondrial dysfunction through the miR-208b/AMPK pathway.
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Ginsenoside Re (Re) is the main component of "Zhenyuan Capsule" (ZYC), which was wildly used in clinic in China for adjunctive treatment of coronary heart disease (CHD) and type II diabetes (T2DM). Nonalcoholic fatty liver disease (NAFLD) is one of the most important complications of T2DM, as well as an important risk factor of CHD. The aim of the present study was to investigate the effects of Re on NAFLD in db/db mice, one of the most recognized gene deficient animal models on T2DM. Sixteen db/db mice and sixteen wild-type mice were divided into four groups and orally administered Re or placebo in equal volume. According to the results, Re showed no obvious effect on blood glucose, lipids, or body weight of db/db mice. Histology pictures of hepatic tissue showed that Re did not improve steatosis, too. However, some evidence suggested that hepatic injury in db/db mice was attenuated by Re administering. Collagen deposition and aminotransferase elevation were significantly downregulated in the DB + Re group compared to those in the DB Group. The mechanisms of the protect effects of Re represented in db/db mice with NAFLD might be inhibiting oxidative stress and the reupregulation of peroxisome proliferator-activated receptor γ (pparγ) expression. The results of this study indicated that ZYC might be able to help T2DM patients with NAFLD to control the progress of NAFLD as an alternation of thiazolidinediones, synthetic agonists of PPARγ, whose side effects and adverse events should not be ignored.
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Since 2015, 170 small molecules, 60 antibody-based entities, 12 peptides, and 15 gene- or cell-therapies have been approved by FDA for diverse disease indications. Recent advancement in medicine is facilitated by identification of new targets and mechanisms of actions, advancement in discovery and development platforms, and the emergence of novel technologies. Early disease detection, precision intervention, and personalized treatments have revolutionized patient care in the last decade. In this review, we provide a comprehensive overview of current and emerging therapeutic modalities developed in the recent years. We focus on nine diseases in three major therapeutics areas, diabetes, autoimmune, and neurological disorders. The pathogenesis of each disease at physiological and molecular levels is discussed and recently approved drugs as well as drugs in the clinic are presented.
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Enfermedades Autoinmunes/terapia , Diabetes Mellitus/terapia , Enfermedades del Sistema Nervioso/terapia , Enfermedades Autoinmunes/patología , Diabetes Mellitus/patología , Humanos , Enfermedades del Sistema Nervioso/patologíaRESUMEN
Around 77 new oncology drugs were approved by the FDA in the past five years; however, most cancers remain untreated. Small molecules and antibodies are dominant therapeutic modalities in oncology. Antibody-drug conjugates, bispecific antibodies, peptides, cell, and gene-therapies are emerging to address the unmet patient need. Advancement in the discovery and development platforms, identification of novel targets, and emergence of new technologies have greatly expanded the treatment options for patients. Here, we provide an overview of various therapeutic modalities and the current treatment options in oncology, and an in-depth discussion of the therapeutics in the preclinical stage for the treatment of breast cancer, lung cancer, and multiple myeloma.
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Anticuerpos Monoclonales/uso terapéutico , Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Terapia Genética/métodos , Inmunoconjugados/uso terapéutico , Neoplasias/terapia , Bibliotecas de Moléculas Pequeñas/uso terapéutico , Humanos , Inmunoterapia/métodos , Inmunoterapia/estadística & datos numéricos , Inmunoterapia/tendencias , Oncología Médica/métodos , Oncología Médica/estadística & datos numéricos , Oncología Médica/tendencias , Terapia Molecular Dirigida/métodos , Terapia Molecular Dirigida/estadística & datos numéricos , Terapia Molecular Dirigida/tendencias , Neoplasias/clasificaciónRESUMEN
With the development of modern chemistry and biology, non-proteinogenic amino acids (NPAAs) have become a powerful tool for developing peptide-based drug candidates. Drug-like properties of peptidic medicines, due to the smaller size and simpler structure compared to large proteins, can be changed fundamentally by introducing NPAAs in its sequence. While peptides composed of natural amino acids can be used as drug candidates, the majority have shown to be less stable in biological conditions. The impact of NPAA incorporation can be extremely beneficial in improving the stability, potency, permeability, and bioavailability of peptide-based therapies. Conversely, undesired effects such as toxicity or immunogenicity should also be considered. The impact of NPAAs in the development of peptide-based therapeutics is reviewed in this article. Further, numerous examples of peptides containing NPAAs are presented to highlight the ongoing development in peptide-based therapeutics.
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Aminoácidos/química , Desarrollo de Medicamentos , Descubrimiento de Drogas , Fragmentos de Péptidos/uso terapéutico , Animales , Humanos , Fragmentos de Péptidos/químicaRESUMEN
Viral infectious diseases have resulted in millions of deaths throughout history and have created a significant public healthcare burden. Tremendous efforts have been placed by the scientific communities, health officials and government organizations to detect, treat, and prevent viral infection. However, the complicated life cycle and rapid genetic mutations of viruses demand continuous development of novel medicines with high efficacy and safety profiles. Peptides provide a promising outlook as a tool to combat the spread and re-emergence of viral infection. This article provides an overview of five viral infectious diseases with high global prevalence: influenza, chronic hepatitis B, acquired immunodeficiency syndrome, severe acute respiratory syndrome, and coronavirus disease 2019. The current and potential peptide-based therapies, vaccines, and diagnostics for each disease are discussed.
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Antivirales/farmacología , Péptidos/farmacología , Vacunas Virales/farmacología , Virosis/tratamiento farmacológico , Síndrome de Inmunodeficiencia Adquirida/diagnóstico , Síndrome de Inmunodeficiencia Adquirida/tratamiento farmacológico , Animales , COVID-19/prevención & control , Vacunas contra la COVID-19/farmacología , Hepatitis B Crónica/diagnóstico , Hepatitis B Crónica/tratamiento farmacológico , Humanos , Vacunas contra la Influenza/farmacología , Gripe Humana/tratamiento farmacológico , Gripe Humana/prevención & control , Síndrome Respiratorio Agudo Grave/diagnóstico , Síndrome Respiratorio Agudo Grave/tratamiento farmacológico , Virosis/prevención & control , Tratamiento Farmacológico de COVID-19RESUMEN
RATIONALE: Hypertrophic cardiomyopathy (HCM) is an inherited myocardial disease and a common cause of sudden cardiac death, heart failure, atrial fibrillation and stroke. In families affected by HCM, genotyping is useful for identifying susceptible relatives. In the present study, we investigated the disease-causing mutations in a three-generation Chinese family with HCM using whole exome sequencing (WES). PATIENT CONCERNS: The proband, a 50-year-old man, was diagnosed with HCM at the age of 41 years. He presented with an asymmetric hypertrophic interventricular septum and a maximum interventricular septum thickness of 18.04âmm. His third elder sister, niece and daughter were also clinically affected by HCM. DIAGNOSIS: Autosomal dominant HCM. INTERVENTIONS: Seven family members, including 4 affected members, accepted WES. The genetic variants were subsequently called using Genome Analysis Toolkit and annotated using the InterVar program. Following frequency filtration by the Genome Aggregation Database, the variants were evaluated using an in-house bioinformatics analysis pipeline. OUTCOMES: HCM was transmitted as an autosomal dominant trait in the family. An extremely rare stop gained mutation, rs796925245 (g.1:201359630G>A, c.835C>T, p.Gln279Ter) in the troponin T2 (TNNT2) gene was identified as the disease-causing mutation. The stop gained mutation was predicted to result in a truncated troponin T protein in cardiac sarcomere. An adolescent family member who had normal echocardiographic measurements was found to carry the same disease-causing mutation. LESSONS: A novel nonsense TNNT2 mutation was identified as the HCM-causing mutation in this Chinese pedigree. Since HCM shows a low penetrance by clinical criteria in adolescents, the adolescent mutation carrier, who is still clinically unaffected, should be offered routine follow-ups and sport activity recommendations to prevent adverse events including sudden cardiac death in the future.
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Cardiomiopatía Hipertrófica Familiar/genética , Troponina T/genética , Adolescente , Adulto , Anciano , Pueblo Asiatico/genética , Cardiomiopatía Hipertrófica Familiar/complicaciones , Cardiomiopatía Hipertrófica Familiar/diagnóstico , Codón sin Sentido , Muerte Súbita Cardíaca/etiología , Ecocardiografía/métodos , Femenino , Humanos , Hipertrofia/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Linaje , Fenotipo , Tabique Interventricular/patología , Secuenciación del Exoma/métodosRESUMEN
The cell membrane is a complex and highly regulated system that is composed of lipid bilayer and proteins. One of the main functions of the cell membrane is the regulation of cell entry. Cell-penetrating peptides (CPPs) are defined as peptides that can cross the plasma membrane and deliver their cargo inside the cell. The uptake of a peptide is determined by its sequence and biophysicochemical properties. At the same time, the uptake mechanism and efficiency are shown to be dependent on local peptide concentration, cell membrane lipid composition, characteristics of the cargo, and experimental methodology, suggesting that a highly efficient CPP in one system might not be as productive in another. To better understand the dependence of CPPs on the experimental system, we present a review of the in vitro assays that have been employed in the literature to evaluate CPPs and CPP-cargos. Our comprehensive review suggests that utilization of orthogonal assays will be more effective for deciphering the true ability of CPPs to translocate through the membrane and enter the cell cytoplasm.
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Membrana Celular/metabolismo , Péptidos de Penetración Celular/química , Péptidos de Penetración Celular/metabolismo , Animales , Membrana Celular/química , Membrana Celular/fisiología , Permeabilidad de la Membrana Celular/fisiología , Péptidos de Penetración Celular/genética , Sistemas de Liberación de Medicamentos/métodos , Endocitosis/fisiología , Humanos , Membrana Dobles de Lípidos/química , Lípidos de la Membrana/química , Lípidos de la Membrana/metabolismo , Transporte de ProteínasRESUMEN
PURPOSE: To accurately distinguish benign from malignant pulmonary nodules with CT based on partial structures of 3D U-Net integrated with Capsule Networks (CapNets) and provide a reference for the early diagnosis of lung cancer. METHOD: The dataset consisted of 1177 samples (benign/malignant: 414/763) from 997 patients provided by collaborating hospital. All nodules were biopsy or surgery proven, and pathologic results were regarded as the "golden standard". This study utilized partial U-Net to capture the low-level (edge, corner, etc.) information and CapNets to preserve high-level (semantic information) information of nodules. For CapNets, each capsule had a 4 × 4 matrix representing the pose and an activation probability representing the presence of an object. Furthermore, we chose accuracy (ACC), area under the curve (AUC), sensitivity (SE) and specificity (SP) to evaluate the generalization of the proposed architecture and compared its identification performance with 3D U-Net and experienced radiologists. RESULTS: The AUC of our architecture (0.84) was superior to that (0.81) of the original 3D U-Net (p = 0.04, DeLong's test). Moreover, ACC (84.5 %) and SE (92.9 %) of our model were clearly higher than radiologists' ACC (81.0 %) and SE (84.3 %) at the optimal operating point. However, SP (70 %) of our model was slightly lower than radiologists' SP (75 %), which might be the result of class imbalance with limited benign samples involved for algorithm training. CONCLUSIONS: Our architecture showed a high performance for identifying benign and malignant pulmonary nodules, indicating the improved model has a promising application in clinic.
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Aprendizaje Profundo , Neoplasias Pulmonares , Nódulo Pulmonar Solitario , Humanos , Neoplasias Pulmonares/diagnóstico por imagen , Interpretación de Imagen Radiográfica Asistida por Computador , Nódulo Pulmonar Solitario/diagnóstico por imagen , Tomografía Computarizada por Rayos XRESUMEN
The progressive loss of cardiomyocytes caused by cell death leads to cardiac dysfunction and heart failure (HF). Rapamycin has been shown to be cardioprotective in pressureoverloaded and ischemic heart diseases by regulating the mechanistic target of rapamycin (mTOR) signaling network. However, the impact of rapamycin on cardiomyocyte death in chronic HF remains undetermined. Therefore, in the current study we addressed this issue using a rat myocardial infarction (MI)induced chronic HF model induced by ligating the coronary artery. Following surgery, rats were randomly divided into six groups, including the sham, vehicle and rapamycinoperated groups, at 8 or 12 weeks postMI. A period of 4 weeks after MI induction, the rats were treated with rapamycin (1.4 mgkgday) or vehicle for 4 weeks. Cardiac function was determined using echocardiography, the rats were subsequently euthanized and myocardial tissues were harvested for histological and biochemical analyses. In the cell culture experiments with H9c2 rat cardiomyocytes, apoptosis was induced using angiotensin II (100 nM; 24 h). Cardiomyocyte apoptosis and autophagy were assessed via measuring apoptosis and autophagyassociated proteins. The activities of mTOR complex 1 (mTORC1) and mTORC2 were evaluated using the phosphorylation states of ribosomal S6 protein and Akt, respectively. The activity of the endoplasmic reticulum (ER) stress pathway was determined using the levels of GRP78, caspase12, phosphoJNK and DDIT3. Echocardiographic and histological measurements indicated that rapamycin treatment improved cardiac function and inhibited cardiac remodeling at 8 weeks postMI. Additionally, rapamycin prevented cardiomyocyte apoptosis and promoted autophagy at 8 weeks postMI. Rapamycin treatment for 4 weeks inhibited the mTOR and ER stress pathways. Furthermore, rapamycin prevented angiotensin IIinduced H9c2 cell apoptosis and promoted autophagy by inhibiting the mTORC1 and ER stress pathways. These results demonstrated that rapamycin reduced cardiomyocyte apoptosis and promoted cardiomyocyte autophagy, by regulating the crosstalk between the mTOR and ER stress pathways in chronic HF.
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Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Insuficiencia Cardíaca/etiología , Insuficiencia Cardíaca/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Transducción de Señal/efectos de los fármacos , Sirolimus/farmacología , Serina-Treonina Quinasas TOR/metabolismo , Angiotensina II/metabolismo , Animales , Línea Celular , Ecocardiografía , Estrés del Retículo Endoplásmico/efectos de los fármacos , Técnica del Anticuerpo Fluorescente , Insuficiencia Cardíaca/diagnóstico , Masculino , RatasRESUMEN
BACKGROUND: Currently, many methodological approaches have been developed to assess peripheral endothelial function. However, a development of the noninvasive and automated technique for routinely assessing endothelial function is still required. We evaluated the potential value of a new method to measure peripheral endothelial function with reactive hyperemia peripheral arterial volume (RH-PAV) in patients with chest pain. METHODS: We used a novel oximeter-like probe to detect the peripheral arterial volume (PAV) of the finger and compared it with brachial flow-mediated dilation (FMD) performed in 93 consecutive patients with chest pain. The RH-PAV index was defined as the ratio of the digital pulse volume during reactive hyperemia relative to the baseline. RESULTS: Ninety-three patients (53 men, 58 ± 5 years) completed the study, and 53 patients demonstrated coronary artery disease (CAD) following scheduled coronary angiography. There was a moderate linear relationship between PAV and FMD (r = 0.69, p < 0.01). Similar to FMD, PAV was more impaired in patients who have more cardiovascular risk factors (CRFs). The subjects with CAD had lower PAV and FMD, compared with those without CAD (1.05 ± 0.23 VS. 1.41 ± 0.37, p < 0.01; 6.7% ± 2.9% VS. 10.4% ± 2.9%, p < 0.01, respectively), and the relationships between FMD and PAV were also significant in both CAD (r = 0.54, p < 0.01) and non-CAD (r = 0.62, p < 0.01) patients. CONCLUSIONS: Endothelial function of digital artery assessed with the novel PAV method demonstrated a profile similar to that of brachial artery measured with FMD. The hyperemia PAV was decreased by factors which were considered to impair endothelial function, suggesting that PAV has the potential to be a novel method to study endothelial function.
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Determinación del Volumen Sanguíneo/métodos , Volumen Sanguíneo , Arteria Braquial/fisiopatología , Enfermedades Cardiovasculares/diagnóstico , Endotelio Vascular/fisiopatología , Dedos/irrigación sanguínea , Fotopletismografía/métodos , Adulto , Anciano , Biomarcadores/sangre , Determinación del Volumen Sanguíneo/instrumentación , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/fisiopatología , Estudios de Factibilidad , Femenino , Hemoglobinas/metabolismo , Humanos , Hiperemia/fisiopatología , Masculino , Persona de Mediana Edad , Fotopletismografía/instrumentación , Proyectos Piloto , Valor Predictivo de las Pruebas , Flujo Pulsátil , Flujo Sanguíneo Regional , VasodilataciónRESUMEN
Ghrelin, a growth hormone-releasing peptide, potentially improves cardiac function, but the mechanisms remain unclear. In the study, the rat heart failure (HF) model was established by ligating the left anterior descending coronary artery (LAD) and treated with ghrelin (100µg/kg, subcutaneous injection, bid); neonatal rat cardiomyocytes were cultured and stimulated with Ang II (0.1µM) and ghrelin(0.1µM) to explore the underlying mechanism of ghrelin in myocardial remodeling. Hemodynamic changes and serum brain natriuretic peptide (BNP) concentrations were measured to assess cardiac function. Left ventricular mass index (LVMI), hematoxylin and eosin (H&E) staining, and Masson's trichrome staining were performed to evaluate myocardial fibrosis. Interestingly, ghrelin significantly improved cardiac function by inhibiting fibrous tissue proliferation. To further explore the mechanisms by which ghrelin interferes with myocardial fibrosis, the levels of activin A (Act A) and its blocker-follistatin (FS) were examined by immunohistochemistry; Act A levels were significantly increased in the myocardial infarction (MI), and ghrelin administeration downregulated Act A expression. In contrast, FS expression showed no significant change in all experimental groups. Furthermore, ghrelin decreased Ang II-induced Act A expression with no effect on FS expression in primary rat cardiomyocytes in vitro (real-time quantitative PCR and ELISA). Thus, ghrelin corrected the Act A/FS imbalance. Finally, Act A treated cultured primary rat cardiac fibroblasts (CFs) showed increased proliferation [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay] and enhanced expressions of type I and type III collagen (Col I and Col III) (real-time quantitative PCR). These data suggest that ghrelin inhibits myocardial fibrosis, attenuates left ventricular remodeling, and eventually improves cardiac function by adjusting Act A/FS imbalance.
Asunto(s)
Folistatina/metabolismo , Ghrelina/farmacología , Insuficiencia Cardíaca , Subunidades beta de Inhibinas/metabolismo , Infarto del Miocardio , Miocardio/metabolismo , Animales , Femenino , Fibrosis , Insuficiencia Cardíaca/tratamiento farmacológico , Insuficiencia Cardíaca/etiología , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/patología , Infarto del Miocardio/complicaciones , Infarto del Miocardio/tratamiento farmacológico , Infarto del Miocardio/metabolismo , Infarto del Miocardio/patología , Miocardio/patología , Ratas , Ratas WistarRESUMEN
Although it is known that the susceptibility of mouse spermatozoa to freezing-thawing varies greatly with genetic background, the underlying mechanisms remain to be elucidated. In this study, to map genetic regions responsible for the susceptibility of spermatozoa to freezing-thawing, we performed in vitro fertilization using spermatozoa from recombinant inbred mice derived from the C57BL/6J and DBA/2J strains, whose spermatozoa showed distinct fertilization abilities after freezing. Genome-wide interval mapping identified two suggestive quantitative trait loci (QTL) associated with fertilization on chromosomes 1 and 11. The strongest QTL on chromosome 11 included 70 genes at 59.237260-61.324742 Mb and another QTL on chromosome 1 included 43 genes at 153.969506-158.217850 Mb. These regions included at least 15 genes involved with testicular expression and possibly with capacitation or sperm motility. Specifically, the Abl2 gene on chromosome 1, which may affect subcellular actin distribution, had polymorphisms between C57BL/6J and DBA/2J that caused at least three amino acid substitutions. A correlation analysis using recombinant inbred strains revealed that the fertilization rate was strongly correlated with the capacitation rate of frozen-thawed spermatozoa after preincubation. This result is consistent with the fact that C57BL/6J frozen-thawed spermatozoa recover their fertilization capacity following treatment with methyl-ß-cyclodextrin to enhance sperm capacitation. Thus, our data provide important clues to the molecular mechanisms underlying cryodamage to mouse spermatozoa.