SUMMARY: Coreopsis tinctoria Nutt. (C. tinctoria Nutt.) can protect diabetic kidneys, but the mechanisms are unclear. This work is to investigate the potential mechanisms of C. tinctoria Nutt. in the treatment of diabetic nephropathy based on network pharmacology analysis of its active ingredients. Twelve small molecular compounds of C. tinctoria Nutt. and targets related to diabetic nephropathy were docked by Discovery Studio 3.0. DAVID database was used for GO enrichment and KEGG pathway analysis. Cytoscape 3.6.1 was used to construct active ingredient-target network. Cell viability was detected with MTT. Glucose consumption was analyzed with glucose oxidase method. Protein expression was measured with Western blot and immunofluorescence. Electron microscopy observed autophagosomes. The core active ingredients of C. tinctoria Nutt. included heriguard, flavanomarein, maritimein, and marein. Twenty-one core targets of the 43 potential targets were PYGM, TLR2, RAF1, PRKAA2, GPR119, INS, CSF2, TNF, IAPP, AKR1B1, GSK3B, SYK, NFKB2, ESR2, CDK2, FGFR1, HTRA1, AMY2A, CAMK4, GCK, and ABL2. These 21 core targets were significantly enriched in 50 signaling pathways. Thirty- four signaling pathways were closely related to diabetic nephropathy, of which the top pathways were PI3K/AKT, insulin, and mTOR, and insulin resistance. The enriched GO terms included biological processes of protein phosphorylation, and the positive regulation of PI3K signaling and cytokine secretion; cellular components of cytosol, extracellular region, and extracellular space; and molecular function of protein kinase activity, ATP binding, and non-membrane spanning protein tyrosine kinase activity. In vitro experiments found that marein increased the expression of phosphorylated AKT/AKT in human renal glomerular endothelial cells of an insulin resistance model induced by high glucose, as well as increased and decreased, respectively, the levels of the microtubule-associated proteins, LC3 and P62. C. tinctoria Nutt. has many active ingredients, with main ingredients of heriguard, flavanomarein, maritimein, and marein, and may exert anti-diabetic nephropathy effect through various signaling pathways and targets.

RESUMEN: Coreopsis tinctoria Nutt. (C. tinctoria Nutt.) puede proteger riñones diabéticos, sin embargo los mecanismos son desconocidos. Este trabajo se realizó para investigar los potenciales mecanismos de C. tinctoria Nutt. en el tratamiento de la nefropatía diabética basado en el análisis de farmacología en red de sus principios activos. Doce compuestos moleculares pequeños de C. tinctoria Nutt. y los objetivos relacionados con la nefropatía diabética fueron acoplados por Discovery Studio 3.0. La base de datos DAVID se utilizó para el enriquecimiento GO y el análisis de la vía KEGG. Se usó Cytoscape 3.6.1 para construir una red de ingrediente-objetivo activa. La viabili- dad celular se detectó mediante MTT. El consumo de glucosa se analizó con el método de glucosa oxidasa. La expresión proteica fue determinada mediante Western blot e inmunofluorescencia. En la microscopía electrónica se observó autofagosomas. Los principales ingredientes activos de C. tinctoria Nutt. incluyeron heriguard, flavanomarein, maritimin y marein. Veintiún de los 43 objetivos potenciales fueron PYGM, TLR2, RAF1, PRKAA2, GPR119, INS, CSF2, TNF, IAPP, AKR1B1, GSK3B, SYK, NFKB2, ESR2, CDK2, FGFR1, HTRA1, AMY2A, CAMK4, GCK y ABL2. Estos 21 objetivos principales se enriquecieron significativamente en 50 vías de señalización. Treinta y cuatro vías de señalización estuvieron estrechamente relacionadas con la nefropatía diabética, de las cuales las principales vías fueron PI3K/ AKT, insulina y mTOR, y resistencia a la insulina. Los términos GO enriquecidos incluyeron procesos biológicos de fosforilación proteica, la regulación positiva de la señalización de PI3K y la secreción de citoquinas; componentes celulares del citosol, región extracelular y espacio extracelular; y la función molecular de la actividad de la proteína quinasa, la unión de ATP y la actividad de la proteína tirosina quinasa que no se extiende por la membrana. Los experimentos in vitro encontraron que la mareína aumentaba la expresión de AKT/AKT fosforilada en células endoteliales glomerulares renales humanas en un modelo de resistencia a la insulina inducida por niveles elevados de glucosa, así como aumentaron y disminuyeron respectivamente, los niveles de las proteínas asociadas a los microtúbulos, LC3 y P62. C. tinctoria Nutt. tiene muchos principios activos, con ingredientes principales de heriguard, flavanomarein, maritimain y marein, y puede ejercer un efecto de nefropatía antidiabética a través de distintass vías de señalización y objetivos.

Abstract Background: Curcuma alismatifolia is an ornamental cultivar with several varieties introduced into China from Thailand within the past fifteen years. Curcuma alismatifolia is widely used as cut flowers and potted flowers and is also used in flower beds. Questions and/or Hypotheses: However, limited genetic and genomic information is available for this species, which has impeded studies on the enhancement of its ornamental value and stress resistance. Studied species Curcuma alismatifolia Study site and dates: Zhejiang, 2018 Methods: single-cell sequencing technology of PCR and polyacrylamide gel electrophoresis Results: (A/T)n accounted for 43.6 % of the SSRs, the maximum proportion, and mononucleotide and trinucleotide repeats were the two most abundant repeat types. A total of 3,637 primer pairs flanking SSR sequences were successfully designed, and 70 sets of primers were randomly selected for validation in 10 varieties. Forty-one (59 %) of the 70 primer pairs successfully amplified alleles, of which 35 were identified as polymorphic markers and used to assess the level of genetic diversity and genetic relationships among the 10 varieties. The genetic diversity analysis showed that the number of alleles (Na) at each locus ranged from 2 to 8, with an average of 3.97, and that PIC had a mean of 0.524 and ranged from 0.095 to 0.795. Conclusions: The genetic distance between 10 varieties varied from 0.30 to 0.96, and the dendrogram clustered all varieties into three groups.

Resumen Antecedentes: Curcuma alismatifolia es un cultivar ornamental con muchasvarias variedades introducidas en China desde Tailandia en los últimos quince años. C. alismatifolia se usa como flor cortada, flor en maceta o para hacer ramilletes de flores. Preguntas y/o Hipótesis: Se dispone de información genética y genómica limitada para esta especie, lo que impide estudios sobre el aumento de su valor ornamental y la resistencia al estrés. Especies estudiadas: Curcuma alismatifolia Lugar de estudio y fechas: Zhejiang, 2018 Métodos: Tecnología de secuenciación unicelular de PCR y electroforesis en gel de poliacrilamida. Resultados: (A/T)n representó el 43.6 % de los SSR, la proporción máxima, y las repeticiones de mononucleótidos y trinucleótidos fueron los dos tipos de repeticiones más abundantes. Se diseñaron con éxito un total de 3,637 pares de cebadores que flanquean las secuencias de SSR y se seleccionaron al azar 70 conjuntos de cebadores para su validación en 10 variedades. 41 (59 %) de los 70 pares de cebadores amplificaron con éxito los alelos, de los cuales 35 se identificaron como marcadores polimórficos y se utilizaron para evaluar el nivel de diversidad genética y las relaciones genéticas entre las 10 variedades. El análisis de diversidad genética mostró que el número de alelos (Na) en cada locus varió de 2 a 8, con un promedio de 3.97, y que el PIC tenía una media de 0.524 y osciló entre 0.095 y 0.795. Conclusiones: La distancia genética entre las 10 variedades varió de 0.30 a 0.96 y en el dendrograma variedades forman tres grupos.

Abstract Cordyceps militaris is a commonly used edible and medicinal fungus in East Asia. Preparation of the mycelium and fermentation liquid by submerged fermentation using additives such as Panax ginseng for skin whitening applications has attracted wide attention. In the present study, we screened traditional Chinese medicines as additives to the liquid fermentation medium for C. militaris and performed systematical research on the changes of bioactivity and ginsenoside levels during the fermentation process. Total polysaccharide contents, cordycepin content, and polysaccharide molecular weight in the fermentation products were measured by UV spectrophotometry, high-performance liquid chromatography-diode array detection (HPLC/DAD), and gel permeation chromatography-multi angle laser light scattering (GPC-MALLS), respectively. In addition, the ginsenosides Rb1, Re, Rd, Rg1, and pseudoginsenoside F11 in the fermentation products were quantitatively analyzed by ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS). Panax ginseng significantly increased the cordycepin and total sugar contents, and the average molecular weight of polysaccharides in the fermentation liquid and the mycelium of C. militaris. At the same time, C. militaris enhanced the production of ginsenoside pseudoginsenoside F11, which is a unique component of Panax quinquefolium. The research provides scientific evidence for the application of C. militaris and P. ginseng as raw materials for food and medicines.

Our objective is to evaluate the accuracy of three algorithms in differentiating the origins of outflow tract ventricular arrhythmias (OTVAs). This study involved 110 consecutive patients with OTVAs for whom a standard 12-lead surface electrocardiogram (ECG) showed typical left bundle branch block morphology with an inferior axis. All the ECG tracings were retrospectively analyzed using the following three recently published ECG algorithms: 1) the transitional zone (TZ) index, 2) the V2 transition ratio, and 3) V2 R wave duration and R/S wave amplitude indices. Considering all patients, the V2 transition ratio had the highest sensitivity (92.3%), while the R wave duration and R/S wave amplitude indices in V2 had the highest specificity (93.9%). The latter finding had a maximal area under the ROC curve of 0.925. In patients with left ventricular (LV) rotation, the V2 transition ratio had the highest sensitivity (94.1%), while the R wave duration and R/S wave amplitude indices in V2 had the highest specificity (87.5%). The former finding had a maximal area under the ROC curve of 0.892. All three published ECG algorithms are effective in differentiating the origin of OTVAs, while the V2 transition ratio, and the V2 R wave duration and R/S wave amplitude indices are the most sensitive and specific algorithms, respectively. Amongst all of the patients, the V2 R wave duration and R/S wave amplitude algorithm had the maximal area under the ROC curve, but in patients with LV rotation the V2 transition ratio algorithm had the maximum area under the ROC curve.

The aim of the present study was to determine the effect of the combination of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and adriamycin (ADM) on the human breast cancer cell line MCF-7 and to identify potential mechanisms of apoptosis. Cell viability was analyzed by the MTT assay and the synergistic effect was assessed by the Webb coefficient. Apoptosis was quantified using the annexin V-FITC and propidium iodide staining flow cytometry. The mRNA expression of TRAIL receptors was measured by RT-PCR. Changes in the quantities of Bax and caspase-9 proteins were determined by Western blot. MCF-7 cells were relatively resistant to TRAIL (IC50 >10 µg/mL), while MCF-7 cells were sensitive to ADM (IC50 <10 µg/mL). A subtoxic concentration of ADM (0.5 µg/mL) combined with 0.1, 1, or 10 µg/mL TRAIL had a synergistic cytotoxic effect on MCF-7 cells, which was more marked with the combination of TRAIL (0.1 µg/mL) and ADM (0.5 µg/mL). In addition, the combined treatment with TRAIL and ADM significantly increased cell apoptosis from 9.8% (TRAIL) or 17% (ADM) to 38.7%, resulting in a synergistic apoptotic effect, which is proposed to be mediated by up-regulation of DR4 and DR5 mRNA expression and increased expression of Bax and caspase-9 proteins. These results suggest that the combination of TRAIL and ADM might be a promising therapy for breast cancer.

We report the structure-activity relationship (SAR) of forty Sansalvamide A (San A) derivatives against human pancreatic ductal adenicarcinoma cell line PL-45. Our comprehensive evaluation of these compounds utilizes: cytotoxicity based SAR, molecular modeling, and ApoAlert Annexin V apoptosis detection to evaluate these potent compounds. Compared to current pancreatic cancer drugs, these San A analogs are structurally unique, and are potentially cytotoxic. Our comprehensive studies including molecular modeling show that a single N-methyl, a single D-amino acid (D-aa) or a single N -methyl D-aa appears to be critical for presenting the active conformation of San A peptide derivates to its biological target, and show that the San A peptide derivative 8 has the ability to selectively induce apoptosis. Thus, showing that this class of compounds are promising chemotherapeutic agents that will eliminate cells in an orderly manner without eliciting an undesired immune response.

Se describe la relación estructura-actividad (REA) de cuarenta derivados de Sansalvamida A (San A) frente a la línea celular PL-45 de adenocarcinoma ductal de páncreas humano. Para la evaluación de estos compuestos se hizo uso de: determinación de la citotoxicidad basada en REA, modelado molecular y detección de apoptosis ApoAlert Annexina V. Comparados a los medicamentos actualmente en uso contra cáncer pancreático, estos análogos de San A de estructura novedosa presentaron actividad citotóxica importante. Nuestros estudios que incluyen modelado molecular muestran que ya sea un solo grupo N-metilo, o un D-aminoácido (D-aa) o un N-metil D-aa pueden ser críticos para presentar la conformación activa del derivado peptídico de San A en su blanco biológico; además muestran que el derivado 8 tiene la habilidad de inducir selectivamente apoptosis. De esta forma, se muestra que esta clase de compuestos son agentes quimioterapéuticos prometedores que eliminan células de manera ordenada, sin presentar una respuesta inmune indeseable.

The balance of body fluids is critical to health and the development of diseases. Although quite a few review papers have shown that several mechanisms, including hormonal and behavioral regulation, play an important role in body fluid homeostasis in adults, there is limited information on the development of regulatory mechanisms for fetal body fluid balance. Hormonal, renal, and behavioral control of body fluids function to some extent in utero. Hormonal mechanisms including the renin-angiotensin system, aldosterone, and vasopressin are involved in modifying fetal renal excretion, reabsorption of sodium and water, and regulation of vascular volume. In utero behavioral changes, such as fetal swallowing, have been suggested to be early functional development in response to dipsogens. Since diseases, such as hypertension, can be traced to fetal origin, it is important to understand the development of fetal regulatory mechanisms for body fluid homeostasis in this early stage of life. This review focuses on fetal hormonal, behavioral, and renal development related to regulation of body fluids in utero.

The role of acetylcholine in the central and peripheral nervous systems is well established in adults. Cholinergic modulation of vascular functions and body fluid balance has been extensively studied. In the embryo-fetus, cholinergic receptors are widespread in the peripheral and central systems, including smooth muscle and the epithelial lining of the cardiovascular, digestive, and urinary systems, as well as in the brain. Fetal nicotine and muscarinic receptors develop in a pattern (e.g., amount and distribution) related to gestational periods. Cholinergic mechanisms have been found to be relatively intact and functional in the control of vascular homeostasis during fetal life in utero at least during the last third of gestation. This review focuses on the development of fetal nicotine and muscarinic receptors, and provides information indicating that central cholinergic systems are well developed in the control of fetal blood pressure and body fluid balance before birth. Therefore, the development of cholinergic systems in utero plays an important role in fetal vascular regulation, gastrointestinal motility, and urinary control.