Exploring the potential of omics technologies in medicinal plant research: a review in Colombia / Explorando el potencial de las tecnologías ómicas en la investigación de plantas medicinales: una revisión en Colombia

This review presents advances in the implementation of high - throughput se quencing and its application to the knowledge of medicinal plants. We conducted a bibliographic search of papers published in PubMed, Science Direct, Google Scholar, Scopus, and Web of Science databases and analyzed the obtained data using VOSviewer (versi on 1.6.19). Given that medicinal plants are a source of specialized metabolites with immense therapeutic values and important pharmacological properties, plant researchers around the world have turned their attention toward them and have begun to examine t hem widely. Recent advances in sequencing technologies have reduced cost and time demands and accelerated medicinal plant research. Such research leverages full genome sequencing, as well as RNA (ribonucleic acid) sequencing and the analysis of the transcr iptome, to identify molecular markers of species and functional genes that control key biological traits, as well as to understand the biosynthetic pathways of bioactive metabolites and regulatory mechanisms of environmental responses. As such, the omics ( e.g., transcriptomics, metabolomics, proteomics, and genomics, among others) have been widely applied within the study of medicinal plants, although their usage in Colombia is still few and, in some areas, scarce. (185)

El extracto de cloroformo (CE) y las fracciones obtenidas de las raíces de Aldama arenaria se evaluaron para determinar su actividad antiproliferativa in vitro contra 10 líneas ce lulares tumorales humanas [leucemia (K - 562), mama (MCF - 7), ovario que expresa un fenotipo resistente a múltiples fármacos (NCI/ADR - RES), melanoma (UACC - 62), pulmón (NCI - H460), próstata (PC - 3), colon (HT29), ovario (OVCAR - 3), glioma (U251) y riñón (786 - 0)]. CE presentó actividad antiproliferativa débil a moderada (log GI 50 medio 1.07), mientras que las fracciones 3 y 4, enriquecidas con diterpenos de tipo pimarane [ent - pimara - 8 (14), ácido 15 - dien - 19 - oico y ent - 8(14),15 - pimaradien - 3 ß - ol], presentaron activid ad moderada a potente para la mayoría de las líneas celulares, con un log GI 50 medio de 0.62 y 0.59, respectivamente. Los resultados mostraron una acción antiproliferativa in vitro prometedora de las muestras obtenidas de A. arenaria , con los mejores resul tados para NCI/ADR - RES, HT29 y OVCAR - 3, y valores de TGI que van desde 5.95 a 28.71 µg.mL - 1, demostrando que los compuestos de esta clase pueden ser prototipos potenciales para el descubrimiento de nuevos agentes terapéuticos

Protective effect of trans-nerolidol on vascular endothelial cell injury induced by lipopolysaccharides / Efectos protectores del trans-nerolidol en lesiones inducidas por lipopolisacáridos en células endoteliales vasculares

This article aimed to discuss the protection of trans - nerolidol on vascular endothelial cells (ECs) injured by lipopolysac charides. ECs were divided into four groups: normal, model, low and high dose trans - nerolidol treatment groups. The cell survival rate and the contents of NO in the cell culture supernatant were determined. The protein expression and transcript level of pe roxisome proliferator - activated receptor - γ (PPARγ), endothelial nitric oxide synthase (eNOS), and inducible nitric oxide synthase (iNOS) were determined by western blotting and RT - PCR respectively. Compared with the normal group, cell livability, protein e xpression and mRNA transcript level of PPARγ and eNOS decreased, NO contents, protein expression and mRNA transcript tlevel of iNOS increased in model group significantly. Compared with model group, all the changes recovered in different degree in treatmen t groups. Hence, it was concluded that trans - nerolidol can alleviate the ECs injuryby the regulation of iNOS/eNOS through activating PPARγ in a dose - dependent manner

Este artículo tiene como objetivo discutir la protección del trans - nerolidol en las células endoteliales vasculares (CE) dañadas por lipopolisacáridos. Las CE se di vidieron en cuatro grupos: normal, modelo, grupos de tratamiento con trans - nerolidol de baja y alta dosis. Se determinó la tasa de supervivencia de las células y los contenidos de óxido nítrico (NO) en el sobrenadante del cultivo celular. La expresión de p roteínas y el nivel de transcripción del receptor activado por proliferadores de peroxisomas - γ (PPARγ), el óxido nítrico sint et asa endotelial (eNOS) y el óxido nítrico sint et asa inducible (iNOS) se determinaron mediante western blot y RT - PCR, respectivamen te. En comparación con el grupo normal, la viabilidad celular, la expresión de proteínas y el nivel de transcripción de PPARγ y eNOS disminuyeron, los contenidos de NO, la expresión de proteínas y el nivel de transcripción de iNOS aumentaron significativam ente en el grupo modelo. En comparación con el grupo modelo, todos los cambios se recuperaron en diferentes grados en los grupos de tratamiento. Por lo tanto, se concluyó que el trans - nerolidol puede aliviar el daño en las CE regulando iNOS/eNOS a través d e la activación de PPARγ de manera dependiente de la dosis.

Diagnóstico prenatal de clorhidrorrea congénita: reporte de caso / Prenatal diagnosis of congenital chloride diarrhea: A case report

La clorhidrorrea congénita es un trastorno genético infrecuente pero importante caracterizado por una alteración grave del balance hidroelectrolítico como resultado de un defecto en la absorción intestinal de cloruros. Los niños afectados presentan diarrea persistente, deshidratación y malnutrición; el control médico y del desarrollo son complejos. Mejorar la detección prenatal es esencial para facilitar la atención del paciente, las intervenciones tempranas y el asesoramiento genético informado. Sin embargo, a pesar de los avances de la medicina, la naturaleza compleja y la escasa frecuencia de esta entidad, constituyen un desafío para la detección prenatal. En este estudio, se reporta el caso de una embarazada donde los estudios por imágenes de resonancia magnética fetales identificaron en forma efectiva las características típicas de la clorhidrorrea congénita. Se proveen conocimientos sobre las complejidades del diagnóstico y se sugieren caminos para las estrategias de detección temprana de esta enfermedad.

Congenital chloride diarrhea (CCD) is a rare but significant genetic disorder characterized by severe electrolyte imbalances resulting from impaired intestinal chloride absorption. Affected children experience persistent diarrhea, dehydration, and malnutrition, complicating medical and developmental care. The enhancement of prenatal detection is crucial for improved patient management, early interventions, and informed genetic counseling. However, despite advancements in medicine, the complex nature and rarity of CCD make prenatal detection challenging. In this study, we report a fetal case where prenatal magnetic resonance imaging (MRI) effectively identified the distinctive characteristics of CCD, providing insights into the complexities of diagnosis and suggesting avenues for enhanced early detection strategies.

Prevalencia y análisis clinicopatológico de pacientes con mutación en el gen PIK3CA con cáncer de mama avanzado Rh+/Her2- en el Instituto Oncológico Nacional De Panamá

Antecedentes: La vía de señalización de la fosfoisitol 3-quinasa (PI3K), que promueve el crecimiento y el metabolismo de las células cancerosas, es la vía mutada con mayor frecuencia en el cáncer de mama y es asociada con quimio resistencia y mal pronóstico. En este estudio presentamos el primer análisis en población panameña y de la región, con ataciones precisas de la mutación PIK3CA, las características clinicopatológicas y pronóstico. Métodos: Estudio exploratorio, donde se recolectaron prospectivamente tumores de 74 pacientes con cáncer de mama metastásico RH+/Her2- del Instituto Oncológico Nacional entre 2022 y 2023. Se realizó un ensayo de PCR en tiempo real para análisis de mutación en ADN extraído del material tumoral fijado en formalina e incluido en parafina para detectar mutaciones en los exones 1, 4, 7, 9 y 20 del gen PIK3CA. Resultados: La mediana de edad de las pacientes estudiadas fue 59 años. La mutación en PIK3CA se encontró en 33.8% (25/74) de pacientes con cáncer de mama, entre ellas 44% fueron mutaciones en el exón 20, 38% en el exón 9, 13% en el exón 4 y 5% en el exón 1. Se observó una correlación significativa entre la mutación y el  tener historia de cáncer en la familia (p= 0.005), y en pacientes postmepáusicas (P = 0.045).  encontramos asociación entre la mutación y el tipo histológico, grado, tamaño tumoral ni estatus axilar al momento del diagnóstico. La mediana de supervivencia libre de progresión  se alcanzó en ambos grupos y tampoco demostró una diferencia significativa. Conclusión: La prevalencia de la mutación es relativamente alta comparada con escenarios internacionales, puede ofrecer una ventaja para elegir las mejores opciones de tratamiento por lo que debe evaluarse de forma rutinaria durante las intervenciones clínicas. (provisto por Infomedic International)

Background: The phosphoisitol 3-kinase (PI3K) signaling pathway, which promotes cancer cell growth and metabolism, is the most frequently mutated pathway in breast cancer and is associated with chemoresistance and poor progsis. In this study we present the first analysis in Panamanian and regional population, with precise antations of the PIK3CA mutation, clinicopathological characteristics and progsis. Methods: Exploratory study, where tumors were prospectively collected from 74 patients with RH+/Her2- metastatic breast cancer from the Instituto Oncológico Nacional between 2022 and 2023. A real-time PCR assay for mutation analysis was performed on DNA extracted from formalin-fixed, paraffin-embedded tumor material to detect mutations in exons 1, 4, 7, 9 and 20 of the PIK3CA gene. Results: The median age of the patients studied was 59 years. The mutation in PIK3CA was found in 33.8% (25/74) of patients with breast cancer, among them 44% were mutations in exon 20, 38% in exon 9, 13% in exon 4 and 5% in exon 1. A significant correlation was observed between the mutation and having  history of cancer in the family (P = 0.005), and in postmepausal patients (P = 0.045). We found  association between the mutation and histologic type, grade, tumor size or axillary status at diagsis. Median progression-free survival was t reached in both groups and did t show a significant difference. Conclusion: The prevalence of the mutation is relatively high compared to international settings, it may offer an advantage in choosing the best treatment options and should be routinely evaluated during clinical interventions. (provided by Infomedic International)

Analysis and 3D Imaging of Morphometric - Mineral Structure Changes in Skulls of Male and Female Guinea Pigs During and After Developmental Period

SUMMARY: Even though morphometric and mineral studies related to the guinea pig (Cavia porcellus) skull have been carried out, this study is the first attempt to evaluate all developmental stages of male and female guinea pigs. This study aims to this study is to create 3D modeling of CT images obtained from the skulls of male and female guinea pigs during the developmental period (prepuberty and the period between puberty and adulthood) and following periods (young adulthood and old adulthood), to analyze some biometric bone data such as volume, surface area and length, and to assess the developmental analysis of the mineral matter change in their skulls. The CT-scanned skulls were transferred to 3D Slicer (5.0.2), which is used for 3D modeling. The surface area and volume were calculated by measuring the measurement points on the models. In addition, the XRF device was used to show elemental ratio changes during different developmental stages. According to metric measurements, a gradual increase was observed during the life period. The metric measurements of the skull bone had a higher measurement value in male guinea pigs than in their female counterparts. While Ca/P ratio increased up to the third group and partially decreased in the fourth group in males, it gradually increased from the first group to the fourth group in females. This study revealed that puberty, adulthood and sex were effective in the physical and chemical characterization of skull bone structure in guinea pigs.

Aunque se han realizado estudios morfométricos y de minerales relacionados con el cráneo del cobayo (Cavia porcellus), esta investigación es el primer intento de evaluar las etapas de desarrollo de cobayos machos y hembras. El objetivo de este estudio fue crear un modelado 3D de imágenes de tomografía computarizada obtenidas de los cráneos de cobayos machos y hembras durante el período de desarrollo (prepubertad y el período entre la pubertad y la edad adulta) y los períodos siguientes (edad adulta joven y edad adulta mayor), para analizar algunos datos biométricos de los huesos, como el volumen, la superficie y la longitud, y además, analizar el cambio de materia mineral en sus cráneos durante el desarrollo. Los cráneos escaneados se transfirieron a 3D Slicer (5.0.2), que se utiliza para el modelado 3D. El área de superficie y el volumen se calcularon midiendo los puntos de medición en los modelos. Además, se utilizó el dispositivo XRF para mostrar los cambios en las proporciones elementales durante diferentes etapas de desarrollo. Según mediciones métricas, se observó un aumento gradual durante el período de vida. Las medidas métricas del hueso del cráneo tuvieron un valor de medición más alto en los cobayos machos que en las hembras. Mientras que la relación Ca/P aumentó hasta el tercer grupo y disminuyó parcialmente en el cuarto grupo en los machos y aumentó gradualmente del primer grupo al cuarto grupo en las hembras. Este estudio reveló que la pubertad, la edad adulta y el sexo fueron efectivos en la caracterización física y química de la estructura ósea del cráneo en cobayos.

The lncRNA-MALAT1/miR-330-3p Axis Promotes Growth and Metastasis of Gastric Cancer Through Regulation of Vascular Endothelial Growth Factor A: A Genetic and Cell Evaluation Study

SUMMARY: Overexpression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in various tumor tissues and cell lines was found to promote tumor cell proliferation, migration, and invasion. However, the role of MALAT1 in gastric cancer (GC) is still unclear. We aimed to investigate the correlation between long-chain non-coding RNAs (lncRNAs), MALAT1, MicroRNAs (miRNA) and vascular endothelial growth factor A (VEGFA) in gastric cancer and to disclose underlying mechanism. The correlation between MALAT1 levels and clinical features was analyzed by bioinformatics data and human samples. The expression of MALAT1 was down regulated in AGS cells to detect the cell proliferation, migration, and invasion characteristics, as well as the effects on signal pathways. Furthermore, we validated the role of MALAT1/miR-330-3p axis in GC by dual luciferase reporter gene assays. Expression of MALAT1 was higher in cancer tissues than in para-cancerous tissues. The high MALAT1 level predicted malignancy and worse prognosis. Down-regulation of MALAT1 expression in AGS cells inhibited cell proliferation, migration, and invasion by targeting VEGFA. By dual luciferase reporter gene assay and miR-330-3p inhibitor treatment, we demonstrate that MALAT1 sponged miR-330-3p in GC, leading to VEGFA upregulation and activation of the mTOR signaling pathway. The MALAT1/miR-330-3p axis regulates VEGFA through the mTOR signaling pathway and promotes the growth and metastasis of gastric cancer.

Se descubrió que la sobreexpresión del transcrito 1 de adenocarcinoma de pulmón asociado a metástasis (MALAT1) en varios tejidos tumorales y líneas celulares promueve la proliferación, migración e invasión de células tumorales. Sin embargo, el papel de MALAT1 en el cáncer gástrico (CG) aún no está claro. Nuestro objetivo fue investigar la correlación entre los ARN no codificantes de cadena larga (lncRNA), MALAT1, los microARN (miARN) y el factor de crecimiento endotelial vascular A (VEGFA) en el cáncer gástrico y revelar el mecanismo subyacente. La correlación entre los niveles de MALAT1 y las características clínicas se analizó mediante datos bioinformáticos y muestras humanas. La expresión de MALAT1 se reguló negativamente en las células AGS para detectar las características de proliferación, migración e invasión celular, así como los efectos sobre las vías de señales. Además, validamos el papel del eje MALAT1/miR- 330-3p en GC mediante ensayos de genes indicadores de luciferasa dual. La expresión de MALAT1 fue mayor en tejidos cancerosos que en tejidos paracancerosos. El alto nivel de MALAT1 predijo malignidad y peor pronóstico. La regulación negativa de la expresión de MALAT1 en células AGS inhibió la proliferación, migración e invasión celular al apuntar a VEGFA. Mediante un ensayo de gen indicador de luciferasa dual y un tratamiento con inhibidor de miR-330-3p, demostramos que MALAT1 esponjaba miR-330-3p en GC, lo que lleva a la regulación positiva de VEGFA y la activación de la vía de señalización mTOR. El eje MALAT1/miR-330-3p regula VEGFA a través de la vía de señalización mTOR y promueve el crecimiento y la metástasis del cáncer gástrico.

Frequência e distribuição do gene da α-actinina-3 e sua associação em atletas brasileiras de voleibol / Frequency and distribution of the α -actinin-3 gene and its association in brazilian volleyball athletes

Introduction: The ACTN3 gene encodes the α-actinin-3 protein in the Z lines of the sarcomere, which anchors the actin protein in the contractile apparatus, present exclusively in type II muscle fibers, presenting greater glycolytic capacity, which is essential for sports with high-energy actions. intensity and short duration as is the case with Volleyball. Objective: To verify the frequency and distribution of the ACTN3 gene, RR and RX genotypes that express α-actinin-3 (EX α-actinin-3), and XX genotype that do not express α-actinin-3 (NE α-actinin-3) and its association with Brazilian volleyball athletes. Materials and Methods: Nine-seven (97) athletes from the women's volleyball super league took part in the study. Body mass, height and age were evaluated to characterize the sample. Salivary samples were analyzed using (PCR) in real time, to determine the genotypes, and, to verify the association of the genotype with the status of volleyball athlete in the three categories (National Teams, Brazilian National Team and Brazilian Olympic Team), the test was carried out Chi-square of independence (χ²). To obtain the odds ratio of the outcome, a log linear regression analysis was performed. All tests were carried out using the JAMOVI 2.4 (2023) statistical software. Results: Among the athletes in the sample competing in the National Teams competition, 91.8% have the EX-α-actinin-3 genotype. When we consider Brazilian National Team competitions, 93.7% have the EX-α-actinin-3 genotype. Athletes who play for the Brazilian Olimpic Team, 100% of the sample have the EX-α-actinin-3 genotype. Considering that in the world population, the frequency is 80%, it is possible to verify that as you approach the athletes who participate in the women's team there is a greater participation of athletes with the EX-α-actinin-3 genotype. Furthermore, there was an association between the genotypes that EX α-actinin-3 and the National category, with the status of elite athlete, where (χ²) obtained the p value (0.023) and the rate ratio (2.71) for the outcome of the genotypes (EX α-actinin-3) being elite athletes. Conclusion: The athlete's genetic characteristics, environment, nutrition, physical, technical and tactical preparation are some of the factors that contribute to sports performance. However, the results of the present study suggest that athletes with RR and RX genotypes that express α-actinin-3, present in type II muscle fibers, seem to confer an advantage when playing high-performance volleyball.

Acondroplasia severa con retraso del desarrollo y acantosis nigricans (SADDAN) / Severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN)

RESUMEN La acondroplasia severa con retraso del desarrollo y acantosis nigricans (SADDAN) es una rara y letal displasia esquelética. Presentamos el primer caso detectado en Perú, en un infante de 13 meses con características fenotípicas de macrocefalia relativa, tórax estrecho, extremidades micromélicas y piel en acordeón; asimismo, un marcado retraso del desarrollo psicomotor en todos los hitos (prueba peruana) y acantosis nigricans. El paciente tuvo mala evolución clínica caracterizada por crisis convulsivas recurrentes, dificultad respiratoria progresiva, y falleció por insuficiencia respiratoria concomitante a neumonía. Esta entidad requiere del acceso a exámenes específicos como el panel de displasias esqueléticas, la cual no es parte de la oferta en la mayoría de los hospitales del Perú. Se requiere una mayor atención las enfermedades raras, a fin de proveer diagnósticos e información oportuna a los involucrados.

ABSTRACT Severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN) is a rare and lethal skeletal dysplasia. We present the first case detected in Peru, in a 13-month-old infant with phenotypic characteristics of relative macrocephaly, narrow thorax, micromelic extremities and accordion skin; likewise, a marked delay in psychomotor development in all milestones (Peruvian test), and acanthosis nigricans. The patient had a poor clinical evolution characterized by recurrent seizures, progressive respiratory difficulty, dying from respiratory failure concomitant to pneumonia. This entity requires access to specific exams such as the skeletal dysplasia panel, which is not part of the offering in most hospitals in Peru. Greater attention is required for rare diseases, to provide timely diagnoses and information to those involved.

Evaluación de Protocolos de Autotrasplante Dental Guiado y sus Tasas de Supervivencia y Éxito. Una Revisión Sistemática

Actualmente los factores que influyen en la supervivencia de los dientes trasplantados han podido ser controlados con el uso de la tecnología. El autotrasplante dental guiado ha logrado más predictibilidad y eficiencia, además, h a reducido los tiempos de transferencia desde la extracción hasta el trasplante. El objetivo de esta revisión es describir los protocolos de autotrasplante dental guiado, sus tasas de supervivencia y éxito publicados en la literatura actual. Esta revisió n fue realizada siguiendo la pauta PRISMA. La búsqueda se realizó en MEDLINE, Google Académico, ScienceDirect y SciELO, con los términos "autotransplant", "autotransplantation", "autotransplanting", "dental", "tooth", "teeth", "guided", "guide" con filtro de publicación de 10 años. Se realizó evaluación de riesgo de sesgo mediante pautas Joanna Briggs Institute (JBI) a los estudios, incluyendo en esta revisión sólo con riesgo medio y bajo. Los datos de cada artículo se tabularon en una tabla realizada en el procesador de texto en línea "Google Docs". Diez estudios cumplieron los criterios mencionados y fueron incluidos. Fueron evaluados un total de 37 pacientes entre 9 a 64 años. Los dientes donantes más frecuentes fueron premolares y terceros molares. En la mayoría de los casos los pacientes se encontraban sin antecedentes mórbidos de relevancia. El éxito fue evaluado mediante diversos criterios clínicos y radiográficos. Por otro lado, la supervivencia fue evaluada durante los seguimientos respecto a la permanencia del diente en boca. Este tipo de tratamiento no es muy conocido y los estudios incluidos fueron escasos, por otro lado, estos son de bajo nivel de evidencia (reportes de casos y serie de casos). Los protocolos evaluados difieren en algunas características, sin embargo, todos logran altas tasas de supervivencia y éxito. Igualmente, se presentan algunos fracasos, dónde los dientes debieron ser extraídos por movilidad e inflamación.

Currently, the factors that influence the survival of transplanted teeth have been controlled with the use of technology. Guided dental autotransplantation has achieved greater predictability and efficiency, and has also reduced transfer times from extraction to transplantation. The aim of this review is to describe the protocols of guided dental autotransplantation, their survival and success rates published in the current literature. This review was performed following the PRISMA guideline. The search was carried out in MEDLINE, Google Scholar, ScienceDirect and SciELO, with the terms "autotransplant", "autotransplantation", "autotransplanting", "autotransplanting", "dental", "tooth", "teeth", "guided", "guide" with a 10-year publication filter. Studies were assessed for risk of bias using Joanna Briggs Institute (JBI) guidelines, including only medium and low risk studies in this review. The data for each article were tabulated in a table created in the online word processor "Google Docs". Ten studies met the selection criteria and were included. A total of 37 patients between 9 and 64 years of age were evaluated. The most frequent donor teeth were premolars and third molars. In most cases the patients had no relevant morbid history. Success was evaluated by means of various clinical and radiographic criteria. On the other hand, survival was evaluated during the follow-ups with respect to the permanence of the tooth in the mouth. This type of treatment is not very well known, and the studies included were scarce; on the other hand, they are of a low level of evidence (case reports and case series). The evaluated protocols differ in some characteristics, however, all of them achieve high survival and success rates. There are also some failures, where teeth had to be extracted due to mobility and inflammation.

Silencing ANGPT2 alleviates ulcerative colitis by regulating autophagy-mediated NLRP3 inflammasome inactivation via the mTOR signaling pathway

Ulcerative colitis (UC) is a difficult intestinal disease characterized by inflammation, and its mechanism is complex and diverse. Angiopoietin-like protein 2 (ANGPT2) plays an important regulatory role in inflammatory diseases. However, the role of ANGPT2 in UC has not been reported so far. After exploring the expression level of ANGPT2 in serum of UC patients, the reaction mechanism of ANGPT2 was investigated in dextran sodium sulfate (DSS)-induced UC mice. After ANGPT2 expression was suppressed, the clinical symptoms and pathological changes of UC mice were detected. Colonic infiltration, oxidative stress, and colonic mucosal barrier in UC mice were evaluated utilizing immunohistochemistry, immunofluorescence, and related kits. Finally, western blot was applied for the estimation of mTOR signaling pathway and NLRP3 inflammasome-related proteins. ANGPT2 silencing improved clinical symptoms and pathological changes, alleviated colonic inflammatory infiltration and oxidative stress, and maintained the colonic mucosal barrier in DSS-induced UC mice. The regulatory effect of ANGPT2 on UC disease might occur by regulating the mTOR signaling pathway and thus affecting autophagy-mediated NLRP3 inflammasome inactivation. ANGPT2 silencing alleviated UC by regulating autophagy-mediated NLRP3 inflammasome inactivation via the mTOR signaling pathway.

Coenzyme Q10 prevents RANKL-induced osteoclastogenesis by promoting autophagy via inactivation of the PI3K/AKT/mTOR and MAPK pathways

Coenzyme Q10 (CoQ10) is a potent antioxidant that is implicated in the inhibition of osteoclastogenesis, but the underlying mechanism has not been determined. We explored the underlying molecular mechanisms involved in this process. RAW264.7 cells received receptor activator of NF-κB ligand (RANKL) and CoQ10, after which the differentiation and viability of osteoclasts were assessed. After the cells were treated with CoQ10 and/or H2O2 and RANKL, the levels of reactive oxygen species (ROS) and proteins involved in the PI3K/AKT/mTOR and MAPK pathways and autophagy were tested. Moreover, after the cells were pretreated with or without inhibitors of the two pathways or with the mitophagy agonist, the levels of autophagy-related proteins and osteoclast markers were measured. CoQ10 significantly decreased the number of TRAP-positive cells and the level of ROS but had no significant impact on cell viability. The relative phosphorylation levels of PI3K, AKT, mTOR, ERK, and p38 were significantly reduced, but the levels of FOXO3/LC3/Beclin1 were significantly augmented. Moreover, the levels of FOXO3/LC3/Beclin1 were significantly increased by the inhibitors and mitophagy agonist, while the levels of osteoclast markers showed the opposite results. Our data showed that CoQ10 prevented RANKL-induced osteoclastogenesis by promoting autophagy via inactivation of the PI3K/AKT/mTOR and MAPK pathways in RAW264.7 cells.

25-hydroxycholesterol aggravates oxygen-glucose deprivation/reoxygenation-induced pyroptosis through promoting activation of NLRP3 inflammasome in H9C2 cardiomyocytes

25-hydroxycholesterol (25-HC) plays a role in the regulation of cell survival and immunity. However, the effect of 25-HC on myocardial ischemia/reperfusion (MI/R) injury remains unknown. Our present study aimed to investigate whether 25-HC aggravated MI/R injury through NLRP3 inflammasome-mediated pyroptosis. The overlapping differentially expressed genes (DEGs) in MI/R were identified from the GSE775, GSE45818, GSE58486, and GSE46395 datasets in Gene Expression Omnibus (GEO) database. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted using the database of Annotation, Visualization and Integration Discovery (DAVID). The protein-protein interaction (PPI) network of the overlapping DEGs was established using the Search Tool for the Retrieval of Interacting Genes (STRING) database. These bioinformatics analyses indicated that cholesterol 25-hydroxylase (CH25H) was one of the crucial genes in MI/R injury. The oxygen-glucose deprivation/reoxygenation (OGD/R) cell model was established to simulate MI/R injury. Western blot and RT-qPCR analysis demonstrated that CH25H was significantly upregulated in OGD/R-stimulated H9C2 cardiomyocytes. Moreover, knockdown of CH25H inhibited the OGD/R-induced pyroptosis and nod-like receptor protein 3 (NLRP3) inflammasome activation, as demonstrated by cell counting kit-8 (CCK8), lactate dehydrogenase (LDH), RT-qPCR, and western blotting assays. Conversely, 25-HC, which is synthesized by CH25H, promoted activation of NLRP3 inflammasome in OGD/R-stimulated H9C2 cardiomyocytes. In addition, the NLRP3 inhibitor BAY11-7082 attenuated 25-HC-induced H9C2 cell injury and pyroptosis under OGD/R condition. In conclusion, 25-HC could aggravate OGD/R-induced pyroptosis through promoting activation of NLRP3 inflammasome in H9C2 cells.

Small Molecules Accelerate Skin Wound Healing: Shikonin Efficacy and Mechanism of Action in Mice / Las Moléculas Pequeñas Aceleran la Curación de Heridas en la Piel: Eficacia de Shikonin y Mecanismo de Acción en Ratones

SUMMARY: The objective of this study was to investigate the therapeutic wound healing potential and molecular mechanisms of shikonin as small molecules in vitro. A mouse burn model was used to explore the potential therapeutic effect of shikonin; we traced proliferating cells in vivo to locate the active area of skin cell proliferation. Through the results of conventional pathological staining, we found that shikonin has a good effect on the treatment of burned skin and promoted the normal distribution of skin keratin at the damaged site. At the same time, shikonin also promoted the proliferation of skin cells at the damaged site; importantly, we found a significant increase in the number of fibroblasts at the damaged site treated with shikonin. Most importantly, shikonin promotes fibroblasts to repair skin wounds by regulating the PI3K/AKT signaling pathway. This study shows that shikonin can effectively promote the proliferation of skin cell, and local injection of fibroblasts in burned skin can play a certain therapeutic role.

El objetivo de este trabajo fue investigar el potencial terapéutico de cicatrización de heridas y los mecanismos moleculares de la shikonina como moléculas pequeñas in vitro. Se utilizó un modelo de quemaduras en ratones para explorar el posible efecto terapéutico de la shikonina; Rastreamos las células en proliferación in vivo para localizar el área activa de proliferación de células de la piel. A través de los resultados de la tinción para patología convencional, encontramos que la shikonina tiene un buen efecto en el tratamiento de la piel quemada y promueve la distribución normal de la queratina de la piel en el sitio dañado. Al mismo tiempo, la shikonina también promovió la proliferación de células de la piel en el sitio dañado. Es importante destacar que encontramos un aumento significativo en la cantidad de fibroblastos en el sitio dañado tratado con shikonina. Lo más importante es que la shikonina promueve la función reparadora de fibroblastos en las heridas de la piel regulando la vía de señalización PI3K/ AKT. Este estudio muestra que la shikonina puede promover eficazmente la proliferación de células de la piel y que la inyección local de fibroblastos en la piel quemada puede desempeñar un cierto papel terapéutico.

Intoxicación severa por 3,4-metilendioximetanfetamina (MDMA). Reporte de caso

La intoxicación por 3,4-metilendioximetanfetamina (MDMA), ha tenido un dramático resurgimiento desde 1980, se ha extendido por gran parte de los Estados Unidos, Europa y América, ha sido ampliamente utilizada como drogas con fines recreativos, actualmente Las catinonas sintéticas se venden como "euforizantes legales" para eludir las leyes existentes, lo que resulta en toxicidad grave y muertes. Presentamos un caso clínico de un adulto joven, quien debuto con intoxicación aguda severa MDMA, con falla multiorgánica, el cual se realizó atención y manejo agudo de la intoxicación en el servicio de urgencias, con posterior manejo de complicaciones en la unidad de cuidados intensivo y finalmente con sobrevida a pesar del mal pronóstico.

3,4-methylenedioxymethamphetamine (MDMA) intoxication, has had a dramatic resurgence since 1980, has spread throughout much of the United States, Europe and America, has been widely used as a recreational drug, currently Synthetic cathinones are they sell as "legal highs" to circumvent existing laws, resulting in severe toxicity and deaths. We present a clinical case of a young adult, who debuted with severe acute MDMA poisoning, with multiple organ failure, who underwent care and acute management of the poisoning in the emergency department, with subsequent management of complications in the intensive care unit and finally with survival despite the poor prognosis.

Expression of miR-210-3p in the aqueous humor of patients with age-related cataracts and its effect on human lens epithelial cell injury induced by hydrogen peroxide / Expressão de miR-210-3p no humor aquoso de pacientes com catarata relacionada à idade e seu efeito na lesão de células epiteliais do cristalino humano induzida por peróxido de hidrogênio

ABSTRACT Purpose: The regulatory effect of microRNA on diseases has been confirmed. This study aimed to evaluate the expression of microRNA-210-3p in age-related cataracts and assess the effect of abnormal miR-210-3p expressions on H2O2-induced SAR01/04 cells. Methods: Reverse-transcription quantitative polymerase chain reaction method was performed to assess the levels of miR-210-3p in aqueous humor samples. Receiver operating characteristic analysis was employed to assess the discrimination ability of miR-210-3p between patients with age-related cataracts and healthy people, and Pearson correlation analysis was used to identify the correlation between miR-210-3p and oxidative stress indices such as superoxide dismutase, glutathione peroxidase, malonaldehyde. Cell counting kit-8 assay and Transwell assay were used to estimate the biological function of H2O2-induced age-related cataract cell model. The levels of oxidative stress indices such as superoxide dismutase, glutathione peroxidase, and malonaldehyde were measured to evaluate the degree of oxidative stress damage in the age-related cataract cell model. The relationship between miR-210-3p and its target gene was verified by luciferase reporter gene analysis. Results: The miR-210-3p expression was elevated in the aqueous humor of patients with age-related cataracts. A high miR-210-3p expression showed a high diagnostic value for age-related cataracts and was significantly associated with the level of oxidative stress markers in patients with age-related cataracts. The inhibition of miR-210-3p can reverse oxidative stress stimulation and adverse effects on H2O2-induced cell function. Conclusions: The results suggested that miR-210-3p could promote cell viability, cell migration, and oxidative stress by targeting autophagy-related gene 7 in in vitro age-related cataract cell model.

RESUMO Objetivo: O efeito regulador do microRNA em doenças tem sido confirmado, e este artigo tentou avaliar a expressão do microRNA-210-3p na catarata relacionada à idade e avaliar o efeito da expressão anormal do miR-210-3p em células SAR01/04 induzidas por H2O2. Métodos: O método de transcrição reversa seguida de reação em cadeia da polimerase (RT-PCR) quantitativa foi realizado para avaliar os níveis de miR-210-3p em amostras de humor aquoso. Análise de características operacionais do receptor foi feita para avaliar a capacidade de discriminação do miR-210-3p entre pacientes com catarata relacionada à idade e pessoas saudáveis. A análise de correlação de Pearson identificou a correlação do miR-210-3p e índices de estresse oxidativo, como superóxido dismutase, glutationa peroxidase, malonaldeído. O ensaio de contagem de células kit-8 (cck-8) e o ensaio no sistema Transwell foram utilizados para estimar a função biológica do formato de células de catarata relacionada com a idade induzida por H2O2. Os níveis de índices de estresse oxidativo como superóxido dismutase, glutationa peroxidase e malonaldeído foram detectados para avaliar o grau de dano do estresse oxidativo em formato de células de catarata relacionada à idade. A relação entre miR-210-3p e seu gene alvo foi verificada por análise do gene repórter luciferase. Resultados: A expressão miR-210-3p foi elevada no humor aquoso de pacientes com catarata relacionada à idade. A expressão miR-210-3p altamente expressiva mostrou alto valor diagnóstico para catarata relacionada à idade e foi significativamente associado ao nível de marcadores de estresse oxidativo em pacientes com catarata relacionada à idade. A inibição de miR-210-3p pode reverter a estimulação do estresse oxidativo e os efeitos adversos da função celular induzida por H2O2. Conclusões: Esses dados sugeriram que a expressão miR-210-3p poderia promover a viabilidade celular, migração celular e estresse oxidativo ao direcionar genes ATG 7 relacionados à autofagia em modelo in vitro de células de catarata relacionadas à idade.

Performance analysis of pretrained convolutional neural network models for ophthalmological disease classification / Análise de desempenho de modelos de rede neural convolucional pré-treinados para classificação de doenças oftalmológicas

ABSTRACT Purpose: This study aimed to evaluate the classification performance of pretrained convolutional neural network models or architectures using fundus image dataset containing eight disease labels. Methods: A publicly available ocular disease intelligent recognition database has been used for the diagnosis of eight diseases. This ocular disease intelligent recognition database has a total of 10,000 fundus images from both eyes of 5,000 patients for the following eight diseases: healthy, diabetic retinopathy, glaucoma, cataract, age-related macular degeneration, hypertension, myopia, and others. Ocular disease classification performances were investigated by constructing three pretrained convolutional neural network architectures including VGG16, Inceptionv3, and ResNet50 models with adaptive moment optimizer. These models were implemented in Google Colab, which made the task straight-forward without spending hours installing the environment and supporting libraries. To evaluate the effectiveness of the models, the dataset was divided into 70%, 10%, and 20% for training, validation, and testing, respectively. For each classification, the training images were augmented to 10,000 fundus images. Results: ResNet50 achieved an accuracy of 97.1%; sensitivity, 78.5%; specificity, 98.5%; and precision, 79.7%, and had the best area under the curve and final score to classify cataract (area under the curve = 0.964, final score = 0.903). By contrast, VGG16 achieved an accuracy of 96.2%; sensitivity, 56.9%; specificity, 99.2%; precision, 84.1%; area under the curve, 0.949; and final score, 0.857. Conclusions: These results demonstrate the ability of the pretrained convolutional neural network architectures to identify ophthalmological diseases from fundus images. ResNet50 can be a good architecture to solve problems in disease detection and classification of glaucoma, cataract, hypertension, and myopia; Inceptionv3 for age-related macular degeneration, and other disease; and VGG16 for normal and diabetic retinopathy.

RESUMO Objetivo: Avaliar o desempenho de classificação de modelos ou arquiteturas de rede neural convolucional pré--treinadas usando um conjunto de dados de imagem de fundo de olho contendo oito rótulos de doenças diferentes. Métodos: Neste artigo, o conjunto de dados de reconhecimento inteligente de doenças oculares publicamente disponível foi usado para o diagnóstico de oito rótulos de doenças diferentes. O banco de dados de reconhecimento inteligente de doenças oculares tem um total de 10.000 imagens de fundo de olho de ambos os olhos de 5.000 pacientes para oito categorias que contêm rótulos saudáveis, retinopatia diabética, glaucoma, catarata, degeneração macular relacionada à idade, hipertensão, miopia, outros. Investigamos o desempenho da classificação de doenças oculares construindo três arquiteturas de rede neural convolucional pré-treinadas diferentes, incluindo os modelos VGG16, Inceptionv3 e ResNet50 com otimizador de Momento Adaptativo. Esses modelos foram implementados no Google Colab o que facilitou a tarefa sem gastar horas instalando o ambiente e suportando bibliotecas. Para avaliar a eficácia dos modelos, o conjunto de dados é dividido em 70% para treinamento, 10% para validação e os 20% restantes utilizados para teste. As imagens de treinamento foram expandidas para 10.000 imagens de fundo de olho para cada tal. Resultados: Observou-se que o modelo ResNet50 alcançou acurácia de 97,1%, sensibilidade de 78,5%, especificidade de 98,5% e precisão de 79,7% e teve a melhor área sob a curva e pontuação final para classificar a categoria da catarata (área sob a curva=0,964, final=0,903). Em contraste, o modelo VGG16 alcançou uma precisão de 96,2%, sensibilidade de 56,9%, especificidade de 99,2% e precisão de 84,1%, área sob a curva 0,949 e pontuação final de 0,857. Conclusão: Esses resultados demonstram a capacidade das arquiteturas de rede neural convolucional pré-treinadas em identificar doenças oftalmológicas a partir de imagens de fundo de olho. ResNet50 pode ser uma boa solução para resolver problemas na detecção e classificação de doenças como glaucoma, catarata, hipertensão e miopia; Inceptionv3 para degeneração macular relacionada à idade e outras doenças; e VGG16 para retinopatia normal e diabética.

Excesso de peso pré-gestacional e ácidos graxos poli-insaturados no leite humano: modelo teórico de causalidade / Pre-gestational overweight and polyunsaturated fatty acids in human milk: theoretical causality model

Resumo Inúmeros estudos têm se detido na avaliação da associação entre o excesso de peso pré-gestacional e os ácidos graxos poli-insaturados no leite humano. Todavia, diante da complexidade de fatores de risco potencialmente confundidores, é recomendável a utilização de ferramentas gráficas para identificar possíveis vieses. O objetivo deste artigo é propor um modelo teórico de causalidade utilizando o gráfico acíclico direcionado entre o excesso de peso pré-gestacional e os ácidos graxos poli-insaturados no leite humano. Foi realizada ampla revisão da literatura para identificar as variáveis com relações causais com a exposição e/ou desfecho. A escolha das variáveis para ajuste seguiu o algoritmo gráfico que compreende seis critérios para a seleção de um conjunto mínimo de variáveis potencialmente confundidoras. Condições socioeconômicas, intervalo interpartal, idade materna e padrão de consumo alimentar foram as variáveis ajustadas a fim de se estimar o efeito total do excesso de peso pré-gestacional sobre o conteúdo dos ácidos graxos poli-insaturados no leite humano. O conjunto mínimo de variáveis encontrado pelo presente estudo pode ser utilizado na análise de outros estudos que avaliem essa associação.

Abstract A number of studies have focused on the evaluation of the relationship between pre-pregnancy overweight and polyunsaturated fatty acids content in human milk. However, given the complexity of potentially confounding risk factors, the use of graphical tools is recommended to identify possible biases. This article aims to propose a theoretical model of causality using the directed acyclic graph between pre-pregnancy overweight and polyunsaturated fatty acids content in human milk. Methods: An extensive literature review was performed to identify variables with causal relationships with exposure and/or outcome. The choice of variables for adjustment followed the graphic algorithm that comprises six criteria for selecting a minimum set of potentially confounding variables. Socioeconomic conditions, interpartum interval, maternal age and food consumption pattern were the variables that would have to be adjusted in order to estimate the total effect of pre-pregnancy overweight on polyunsaturated fatty acids content in human milk. The minimum set of variables found in the present study can be used in the analysis of other studies that evaluate this association.

Standardized Centella asiatica extract ECa 233 alleviates pain hypersensitivity by modulating P2X3 in trigeminal neuropathic pain

Abstract During oral surgery and temporomandibular joint repositioning, pain hypersensitivity often occurs due to irritation or inflammation of the nerve endings in the orofacial region. Objective: This study aimed to investigate the effects of ECa 233, a Centella asiatica-standardized extract, on the development of mechanical hyperalgesia and allodynia induced by chronic constriction injury of the infraorbital nerve in mice. Methodology: The right infraorbital nerves of the mice were ligated. Oral carbamazepine (20 mg/kg) or ECa 233 (30, 100, or 300 mg/kg) was administered daily for 21 days. Von Frey and air-puff tests were performed on both sides of the whisker pad on days 0, 7, 14, and 21. Thereafter, the expression of purinergic receptor subtype 3 (P2X3) and voltage-gated sodium channel 1.7 (NaV1.7), a transmembrane protein, in the trigeminal ganglion and c-fos immunoreactivity-positive neurons in the trigeminal nucleus caudalis was assessed. Results: After 21 days of infraorbital nerve ligation, the mice showed allodynia- and hyperalgesia-like behavior, P2X3 and NaV1.7 were upregulated in the trigeminal ganglion, and nociceptive activity increased in the trigeminal nucleus caudalis. However, the oral administration of carbamazepine (20 mg/kg), ECa 233 (100 mg/kg), or ECa 233 (300 mg/kg) mitigated these effects. Nevertheless, ECa 233 failed to affect NaV1.7 protein expression. Conclusion: Carbamazepine and ECa 233 can prevent pain hypersensitivity in mice. Considering the side effects of the long-term use of carbamazepine, ECa 233 monotherapy or combined ECa 233 and carbamazepine therapy can be used as an alternative for regulating the development of hypersensitivity in trigeminal pain. However, further detailed clinical studies should be conducted to provide comprehensive information on the use of ECa 233.

Insulin-like growth factor binding protein 3 promoter variant (rs2854744) is associated with nonalcoholic fatty liver disease

ABSTRACT Objective: Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease and a growing global epidemic. In NAFLD, liver fat surpasses 5% of hepatocytes without the secondary causes of lipid accumulation or excessive alcohol consumption. Given the link between NAFLD and insulin resistance, the possible association between the rs2854744 (−202 G>T) promoter polymorphism of insulin-like growth factor binding protein 3 (IGFBP3) gene and NAFLD was investigated in this study. Materials and methods: In this genetic case-control association study, the IGFBP3 rs2854744 genotypes of 315 unrelated individuals, including 156 patients with biopsy-proven NAFLD and 159 controls, were determined using polymerase chain reaction/restriction fragment length polymorphism analyses. Results: The "GT+TT" genotype of the IGFBP3 rs2854744 polymorphism, compared with the "GG" genotype, was associated with a 2.7-fold increased risk of NAFLD after adjustment for confounding factors (P = 0.009; odds ratio [OR] = 2.71; 95% confidence interval [CI] = 1.19-3.18). Additionally, the IGFBP3 rs2854744 "T" allele, in comparison with the "G" allele, was significantly overrepresented in NAFLD patients than the controls (P = 0.008; OR = 1.85; 95%CI = 1.23-2.94). Conclusion: Our findings first indicated that the IGFBP3 rs2854744 "GT+TT" genotype is a marker of increased NAFLD susceptibility; however, it needs to be supported by further investigations in other populations.

Hsa_circ_0013561 promotes progression of nasopharyngeal carcinoma by activating JAK2/STAT3 signaling pathway

Abstract Objective Nasopharyngeal Carcinoma (NPC) is a malignancy of epithelium of epithelium of the nasopharynx, with the highest incidence of otolaryngeal malignancies. A growing number of studies confirm that Circular RNA (circRNA) plays an important role in tumor development, including Hsa_circ_0013561. This study aims to elucidate the process and mechanism of NPC regulation hsa_circ_0013561. Methods In this study, circRNA expression nodes and subcellular localization in NPC tissues were analyzed by fluorescence in situ hybridization. The expression of hsa_circ_0013561 in NPC cells was further clarified by RT-qPCR. At the same time, the lentivirus vector interfered by hsa_circ_0013561 was constructed and transfected. The cell proliferation was detected by CCK-8 method, EdU assay and plate cloning assay. The cell cycle and apoptosis were detected by flow cytometry, and the cell migration ability was detected by wound healing assay and Transwell assay. Western blotting examined the expression of apoptosis, Epithelial-Mesenchymal Transition (EMT)-associated proteins, and Janus Kinase/Signal Transductor and Activator of Transcription (JAK/STAT) signaling pathway-related proteins. Results The results showed that the expression of hsa_circ_0013561 in NPC samples was significantly upregulated and hsa_circ_0013561 localized in the cytoplasm. After down-regulating hsa_circ_0013561 expression, it significantly inhibited the proliferation and metastasis ability of NPC, inhibited EMT progression, and promoted apoptosis. Further studies showed that interference hsa_circ_0013561 significantly inhibited JAK2/STAT3 signaling pathway activation and induced the expression of apoptosis-related proteins. Conclusion In summary, we found that hsa_circ_0013561 is a pro-tumor circRNA in NPC, which can reduce the activation of JAK2/STAT3 pathway by knocking down hsa_circ_0013561, thereby slowing down the malignant progression of NPC. Oxford Centre for Evidence-Based Medicine 2011 Levels of Evidence Level 4.