Screening and characterization estrogen receptor ligands from Arnebia euchroma (Royle) Johnst. via affinity ultrafiltration LC-MS and molecular docking.

Arnebiae Radix (dried root of Arnebia euchroma (Royle) Johnst.) is a traditional Chinese medicine (TCM) used to treat macular eruptions, measles, sore throat, carbuncles, burns, skin ulcers, and inflammations. The Arnebiae Radix extract can exert anti-breast cancer effects through various mechanisms of action. This study aimed to rapidly screen potential estrogen receptor (estrogen receptor α and estrogen receptor ß) ligands from the Arnebiae Radix extract. In this study, an analytical method based on affinity ultrafiltration coupled with UHPLC-Q-Exactive Orbitrap mass spectrometry was established for rapidly screening and identifying estrogen receptor ligands. Then, bindings of the components to the active site of estrogen receptor (estrogen receptor α and estrogen receptor ß) were investigated via molecular docking. Moreover, surface plasmon resonance (SPR) experiments with six compounds were performed to verify the affinity. As a result, a total of 21 ligands were screened from Arnebiae Radix using affinity ultrafiltration. Among them, 14 and 10 compounds from Arnebiae Radix showed affinity with estrogen receptor α and estrogen receptor ß, respectively. All of those ligands could have a good affinity for the multiple amino acid residues of the estrogen receptor based on molecular docking. In addition, six compounds display the great affinity by SPR. The method established in the study could be used to rapidly screen estrogen receptor ligands in Traditional Chinese medicine. The results demonstrated that the affinity ultrafiltration-UHPLC-Q-Exactive Orbitrap mass spectrometry method not only aids in the interpretation of the potential bioactive components and possible mechanisms of action of Arnebiae Radix but also provides a further effective basis for the quality control of this valuable herb medicine.

A Systematic Method for the Identification of Aporphine Alkaloid Constituents in Sabia schumanniana Diels Using UHPLC-Q-Exactive Orbitrap/Mass Spectrometry

Sabia schumanniana Diels (SSD) is a plant whose stems are used in traditional folk medicine for the treatment of lumbago and arthralgia. Previous studies have revealed chemical constituents of SSD, including triterpenoids and aporphine alkaloids. Aporphine alkaloids contain a variety of active components, which might facilitate the effective treatment of lumbago and arthralgia. However, only 5-oxoaporphine (fuseine) has been discovered in SSD to date. In this study, we sought to systematically identify the aporphine alkaloids in SSD. We established a fast and reliable method for the detection and identification of these aporphine alkaloids based on ultra-high-performance liquid chromatography (UHPLC)-Q-Exactive-Orbitrap/mass spectrometry combined with parallel reaction monitoring (PRM). We separated all of the analyzed samples using a Thermo Scientific Hypersil GOLD™ aQ C18 column (100 mm × 2.1 mm, 1.9 µm). Finally, we identified a total of 70 compounds by using data such as retention times and diagnostic ions. No fewer than 69 of these SSD aporphine alkaloids have been reported here for the first time. These findings may assist in future studies concerning this plant and will ultimately contribute to the research and development of new drugs.

A prospective study of total urinary deoxynivalenol in adolescents in Shanghai, China.

There is growing interest in deoxynivalenol (DON) exposure during puberty because experimental evidence shows that DON-exposed to adolescents are more sensitive to DON and have limited detoxification ability. Nevertheless, there have been few surveys of DON exposure for adolescents in China. Furthermore, little is known about the effects of collection times on risk exposure. In the study, we estimated the risk of DON exposure for adolescents in Shanghai and explore the effects of collection time, areas, sex and BMI on intake estimates. A total of 315 adolescents aged 14-16 years, including 161 boys and 154 girls, were recruited. Urine samples were collected for three consecutive days and digested with ß-glucuronidase and then purified using a DON-immunoaffinity column (IAC). Total deoxynivalenol levels were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in combination with a stable isotope dilution assay (SIDA). DON was detected in 945 morning urine samples taken from 315 individuals for three consecutive days. The mean estimated dietary intake of DON did not exceed the tolerable daily intake (TDI) of l µg/kg·bw/day, showing that exposure of adolescents in Shanghai is not of concern. However, 10-20% of probable daily intake (PDI) values exceed the TDI, indicating potential adverse effects. In addition, the DON concentration at the population level did not differ for urine samples collected at different times except for those of overweight adolescents. Therefore, assessment of exposure to DON by monitoring the morning urine of a healthy adolescent, except for overweight people, provides an appropriate estimate of exposure and related risk at the population level, but intake estimates for individuals are uncertain; these could be used to assess exposure of adolescents to DON rapidly and effectively for epidemiological investigations.

Identification of Metabolites of Aurantio-Obtusin in Rats Using Ultra-High-Performance Liquid Chromatography-Q-Exactive Orbitrap Mass Spectrometry with Parallel Reaction Monitoring.

Aurantio-obtusin (AO) is a major anthraquinone compound isolated from Cassiae Semen or Duhaldea nervosa, which possesses diverse pharmacological effects. Previous studies have shown that it has a good effect on lowering blood lipids and treating various diseases. A few studies have also reported about its metabolites. A rapid and reliable method using ultra-high-performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry and multiple data-processing technologies was established to investigate the metabolites of AO in the plasma and various tissues of rats, including the heart, liver, spleen, lung, kidneys, and brain. Finally, a total of 36 metabolites were identified in the plasma of rats, which could be very beneficial for understanding the effective form of AO metabolites leading to new drug discovery. The result demonstrated that this strategy, especially parallel reaction monitoring, has shown a wide range of applications in the identification of metabolites.

miR-1293 Suppresses Tumor Malignancy by Targeting Hydrocyanic Oxidase 2: Therapeutic Potential of a miR-1293/Hydrocyanic Oxidase 2 Axis in Renal Cell Carcinoma.

Renal cell carcinoma (RCC) is a common cancer, and extensive research suggests that microRNA may play an important role in the progression of RCC. The emphasis of this article was to reveal the function and mechanism of microRNA-1293(miR-1293) in the development of RCC tumors. First, the authors carried out bioinformatics analysis. The differential expression of miR-1293 in RCC tumor and normal cells was analyzed using the data from The Cancer Genome Atlas database, and Kaplan-Meier survival analysis was carried out to test the survival rate. Subsequently, the miR-1293 expression in RCC cell lines was examined by quantitative real-time PCR. Then Cell counting kit-8 and Transwell assays were executed to detect the function of miR-1293 in RCC. Bioinformatics prediction, western blotting, and dual-luciferase reporter assay were set to check the target gene of miR-1293. Finally, they conducted rescue experiments to verify whether the regulation of miR-1293 on the biological function of RCC cells was achieved by regulating hydrocyanic oxidase 2 (HAO2). Bioinformatics results showed that miR-1293 was highly expressed in RCC, and the miR-1293 high-expression group showed a lower survival rate than the miR-1293 low-expression group, which suggested that the high expression of miR-1293 was related to unfavorable prognosis in RCC. Subsequent assays evidenced that upregulation of miR-1293 expression significantly increased the cell viability and promoted cell migration and invasion in RCC. Silencing miR-1293 expression showed opposite results. Furthermore, HAO2 was confirmed to be a direct target gene of miR-1293 by dual-luciferase reporter assay, and miR-1293 negatively regulated the expression of HAO2. Moreover, rescue experiments evidenced that miR-1293 reduced the cell viability, invasion, and migration of RCC by regulating HAO2. In sum, miR-1293 can regulate the viability, invasion, and migration of RCC tumor cells by targeting HAO2, suggesting that miR-1293 can be used as a new biomarker for clinical treatment of RCC.

Fabrication of FeOx-Decorated on Ethanol Pretreated Ti Grids for Asymmetric Supercapacitors.

FeOx grown on titanium grids were synthesized by a hydrothermal approach and their electrochemical properties were investigated. Among all FeOx materials, FeOOH-decorated Ti grid with ethanol dried (Ti@FeOOH-1) exhibits a higher specific capacitance (159.7 F g-1 at a current density of 0.5 A g-1) than that of Fe2O3-decorated Ti grid with ethanol dried (Ti@Fe2O3-1) (51.9 F g-1 at a current density of 0.5 A g-1). However, the capacitance retention of Ti@Fe2O3-1 after 1000 cycles is about 83.3% of the initial capacitance while Ti@FeOOH-1 only keeps cycling performance with 67.7% retention. Considering the importance of high specific capacitance for supercapacitor, we chose Ti@FeOOH-1 as the electrode material in our experiment. An asymmetric supercapacitor was successfully fabricated using Ti@MnO2 as the positive electrode and Ti@FeOOH-1 as the negative electrode. The tests show that the asymmetric supercapacitor has high specific capacitance (77 F g-1 at a current density of 1 A g-1) and excellent cycling ability (97.5% retention after 1000 cycles). The synthesis of FeOx grown on titanium grids brings new probability to design a potential negative electrode material for supercapacitors.

Tuning Hierarchical Ferric Nanostructures-Decorated Diatomite for Supercapacitors.

FeOOH nanosheets on porous diatomite have been successfully prepared by a facile two-step hydrothermal approach for supercapacitors, and then α-Fe2O3 and γ-Fe2O3 nanostructures are obtained via calcination under different atmospheres and temperatures. The morphologies and structures of all the samples are investigated in detail to make the hierarchical architecture clear. Besides, systemic tests are carried out in 1 M Na2SO4 electrolyte to characterize the electrochemical properties of these materials. Among the iron-related composite electrodes, diatomite@FeOOH owns the highest specific capacitance (157.9 F g-1 at a current density of 0.5 A g-1) and best cycling performance (98.95% retention after 1000 cycles), which is considered to be a potential material for high-performance supercapacitors. Furthermore, the synthesizing strategy can be extended to the preparation of other metallic oxide-derived functional materials towards energy storage and conversion.

Protective effects of BMSCs in combination with erythropoietin in bronchopulmonary dysplasia-induced lung injury.

Bronchopulmonary dysplasia (BPD) is the most common type of chronic lung disease in infancy, for which no effective therapy is currently available. The aim of the present study was to investigate the effect of treatment with bone marrow mesenchymal stem cells (BMSCs) in combination with recombinant human erythropoietin (rHuEPO) on BPD­induced mouse lung injury, and discuss the underlying mechanism. The BPD model was established by the exposure of neonatal mice to continuous high oxygen exposure for 14 days, following which 1x106 BMSCs and 5,000 U/kg rHuEPO were injected into the mice 1 h prior to and 7 days following exposure to hyperoxia. The animals received four treatments in total (n=10 in each group). After 14 days, the body weights, airway structure, and levels of matrix metalloproteinase­9 (MMP­9) and vascular endothelial growth factor (VEGF) were detected using histological and immunohistochemical analyses. The effect on cell differentiation was observed by examining the presence of platelet endothelial cell adhesion molecule (PECAM) and VEGF using immunofluorescence. Compared with the administration of BMSCs alone, the body weight, airway structure, and the levels of MMP­9 and VEGF were significantly improved in the BMSCs/rHuEPO group. The results of the present study demonstrated that the intravenous injection of BMSCs significantly improved lung damage in the hyperoxia­exposed neonatal mouse model. Furthermore, the injection of BMSCs in combination with intraperitoneal injection of rHuEPO had a more marked effect, compared with BMSCs alone, and the mechanism may be mediated by the promoting effects of BMSCs and EPO. The results of the present study provided information, which may assist in future clinical trials.

siRNA Suppression of NEDD9 Inhibits Proliferation and Enhances Apoptosis in Renal Cell Carcinoma.

Renal cell carcinoma (RCC) is the most lethal of all genitourinary malignancies. NEDD9/HEF1/Cas-L is a member of the Cas protein family and is known as a biomarker in multiple cancer types. In this study, we demonstrate for the first time that NEDD9 was upregulated in RCC tissue and cell lines. Immunohistochemical analysis and quantitative RT-PCR analysis showed low expression of NEDD9 in normal renal tissues and high expression in RCC tissues. In addition, in vitro experiments show that expression of NEDD9 was upregulated in RCC cell lines. Through MTT assay, we observed that NEDD9 knockdown inhibited cell proliferation. Furthermore, flow cytometry analysis showed that NEDD9 downregulation induced apoptosis. Together, our data suggest that abnormal NEDD9 protein expression may be a marker for RCC, and NEDD9 knockdown suppresses cell growth.

The Presence of Intramitochondrial Yolk-crystals in Oocytes of Hypophysectomized Bullfrog Tadpoles*: (intramitochondrial yolk-crystal/tadpole oocyte (Rana)/hypophysectomized tadpole).

Ovaries of hypophysectomized Rana catesbeiana tadpoles. weighing I to 14 g, were prepared for electron microscopic study. The oocytes are at the growth phase, ranging from 50 to 190 µm in diameter. The observation on these oocytes has revealed the presence of intramitochondrial yolk-crystals but not cytoplasmic yolk platelets. The crystalline structure, situated within the intracristal space, consists of a hexagonal array of dense particles about 50 Å in diameter and 72 Å in periodicity. Our data agree with those reported in oocytes of intact ranid species. According to literatures, crystals of intramitochondrial yolk and of cytoplasmic yolk platelets show similar ultrasturctures. The precursor of cytoplasmic yolk platelets in adult Xenopus oocytes is known to be synthesized in the estrogen-stimulated liver and incorporated via circulation into oocytes by gonadotropin-dependent micropinocytosis. The present finding suggests that the intramitochondrial yolk could be formed within oocytes, independently of the pituitary control.