Screening and validation of lymph node metastasis risk-factor genes in papillary thyroid carcinoma.

Background: Although most papillary thyroid carcinoma (PTC) cases have a good prognosis, some PTCs are more aggressive and are often accompanied by lymph node (LN) metastasis, a high recurrence rate, and poor prognosis. Distinguishing highly invasive metastatic PTC is an urgent problem that needs to be addressed clinically. We analyzed a microarray of metastasized PTC and validated it using quantitative reverse transcription PCR (RT-qPCR) and immunohistochemistry to identify biomarkers that can be used to assess the risk of PTC metastasis. Methods: The microarray of metastasized PTC was screened using the Gene Expression Omnibus (GEO) database. The differences between cancer and normal tissues were analyzed using the official GEO tool: GEO2R. Gene expression profile data (GEPIA) were used to verify the expression of differential genes in large samples and to analyze their correlation. The Kaplan-Meier plotter (KM-plotter) database was used for the analysis of genes potentially related to survival. RT-qPCR was used to check the expression of risk factor genes in pathological sections from PTC patients with clinical LN metastasis. Immunohistochemistry was used to verify the expression of core risk-associated genes. Results: Fourteen PTC metastasis-associated genes were identified. In metastasized PTC, CLDN1, LRP4, LRRK2, and TENM1 were highly expressed, whereas DIO1, DPP6, HGD, IPCEF1, MT1F, SLC26A4, SLC26A7, SPX, TFF3, and TPO were expressed at low levels, compared to expression in normal tissues. DIO1, HGD, SLC26A4, and TPO were found to be the core risk genes in the PTC metastatic risk set. Results based on clinical samples showed that the expression differences for metastasis risk-associated genes were consistent with the bioinformatics analysis results. Conclusions: Fourteen differentially expressed genes (CLDN1, LRP4, LRRK2, TENM1, DIO1, DPP6, HGD, IPCEF1, MT1F, SLC26A4, SLC26A7, SPX, TFF3, TPO) are associated with an increased risk of PTC metastasis, and DIO1, HGD, SLC26A4, and TPO are the key risk-associated genes in this set that might affect the occurrence and development of PTC through iodine metabolism. These genes could provide a reference for clinical metastatic PTC risk evaluation and treatment.

Water-extracted Lonicera japonica polysaccharide attenuates allergic rhinitis by regulating NLRP3-IL-17 signaling axis.

In this study, the immunomodulatory effects and mechanism of action of a novel water-extracted Lonicera japonica polysaccharide (WLJP) on allergic rhinitis (AR) was investigated. For the efficacy of WLJP, behavioral symptoms (rubbing and sneezing), serum inflammatory factors, pathological damage, splenic T cell differentiation, gut microbiota imbalance, and protein analysis of the nasal mucosa and colon were assessed. WLJP and the NLRP3 inhibitor, CY-09, were co-evaluated in the AR model established using LPS + IFN-γ-induced THP-1 cells. The WLJP group showed decreased serum inflammatory factors, eosinophils, goblet cells, NLRP3 inflammasomes, splenic Th17 cell differentiation, and expression of IL-17, p-p65, and gut NLRP3 in the nasal mucosa while maintaining gut microbiota balance, repairing the mechanical barrier, and significantly improving AR behavioral symptoms. In vitro interaction analysis showed a significant interaction between CY-09 and WLJP. In conclusion, WLJP improves AR by repairing the gut barrier and inhibiting NLRP3 inflammasome-driven inflammation and the Th17 immune response.

Inhibition of reinstatement of alcohol-induced conditioned place preference in mice by Lonicera japonica polysaccharide.

Alcohol is one of the most commonly used addictive substances. Addiction memory is a necessary part of the mechanism underlying drug addiction. Lonicera japonica and its extract can mitigate organ damage caused by alcohol. In this study, Lonicera japonica polysaccharide (LJP) was extracted, and its effect on alcohol addiction memory was investigated. LJP inhibited the cue-induced reinstatement of conditioned place preference and elevated Tuj1 and DCX levels in the hippocampus to suppress neuronal damage. LJP also reduced the hippocampal glutamate (Glu) levels, alcohol-induced abnormal enhancement of the addiction memory, and decreased VPS34 phosphorylation and hyper activation of autophagy pathways in the hippocampus of alcohol-dependent mice. Our results suggest that LJP is a potential functional food to treat or ameliorate alcohol-induced addiction and that VPS34 is a potential target for modulating alcohol cravings.

Research progress on the protective effects of aucubin in neurological diseases.

CONTEXT: Aucubin (AU), an iridoid glycoside that is one of the active constituents of Eucommia ulmoides Oliv. (EUO) (Eucommiaceae), a traditional Chinese medicine, has been extensively studied in the management of neurological diseases (NDs). However, a comprehensive review of its effects and mechanisms in this regard is currently not available. OBJECTIVE: To compile the protective effects and mechanisms of AU in NDs and provide a basis for further research. METHODS: We used 'aucubin' as the 'All Fields' or 'MeSH' in PubMed, Web of Science and China National Knowledge Infrastructure without any limitation to search all relevant articles as comprehensively as possible; we selected the articles on AU treatment of NDs for summary. RESULTS: Studies reviewed herein reported that AU improved the symptoms or prognosis of Parkinson's disease, Alzheimer's disease, intracerebral haemorrhage, diabetic encephalopathy, epilepsy, anxiety and depression, and traumatic brain injury. The pharmacological mechanisms involved in repairing neuronal loss were postulated to include increasing γ-aminobutyric acid (GABA) content in the synapse, promoting differentiation of neural precursor cells into GABAergic neurons, providing antioxidant and anti-neuroinflammation activities, as well as enhancing autophagy and anti-apoptotic actions. DISCUSSION AND CONCLUSIONS: The protective effects of AU on some NDs have been confirmed. According to the pharmacological effects, AU is also highly likely to have protective effects on other NDs, which can be realized by further in vivo and in vitro basic research, and clinical trials. In the future, AU may be used for clinical prevention or treatment of patients with neurological diseases.

Tailoring surface morphology of heterostructured iron-based Fenton catalyst for highly improved catalytic activity.

Due to the great limitation of glass forming ability, precisely controlling the chemical compositions of metallic glasses (MGs) still dramatically inhibits their widespread applications in wastewater remediation. Here, heterostructured catalysts were exploited by rapid annealing of Fe-based MGs and subsequent ball milling (BM) as advanced alternatives for amorphous counterparts in Fenton-like process. It was found that the surface characteristics tailored by ball milling enable more chemically active sites due to its enlarged specific surface area, surface defects and nanosized amorphous oxide layer that significantly enhance surface-catalyzed reaction in Fenton-like process. On the other hand, high-temperature annealing induced grain growth and electrochemical potential difference induced effect of galvanic cells in multiple crystalline phases (e.g. α-Fe (Si), Fe2B and Fe3Si) further provide an important contribution to high efficiency of electron transfer in heterostructured catalysts. Since the multiphase heterostructure is easily formed by a high-temperature annealing of MGs/amorphous-crystalline composite alloys, this work aims to provide an advanced alternative of MG catalyst without the elemental limitation of glass forming ability for wastewater remediation.

Comparison of a New Optical Biometer That Combines Scheimpflug Imaging With Partial Coherence Interferometry With That of an Optical Biometer Based on Swept-Source Optical Coherence Tomography and Placido-Disk Topography.

PURPOSE: To evaluate measurement precision and to compare the Pentacam AXL (Oculus Optikgeräte, Wetzlar, German), a new optical biometer based on Scheimpflug imaging and partial coherence interferometry (PCI) with that of the OA-2000 biometer (Tomey, Nagoya, Japan), which combines swept-source optical coherence tomography (SS-OCT) and Placido-disk topography. METHODS: Axial length (AL), central corneal thickness (CCT), anterior chamber depth (ACD), aqueous depth (AQD), mean keratometry (Km), astigmatism vectors J0, J45, and corneal diameter (CD) were measured in triplicate by two technical operators. Within-subject standard deviation (Sw), repeatability and reproducibility (2.77 Sw), coefficient of variation (CoV), and intraclass correlation coefficient (ICC) were used to assess the Pentacam AXL intra-observer repeatability and inter-observer reproducibility. Paired t-test and Bland-Altman plots were used to determine the agreement between the two biometers. RESULTS: The new optical biometer had high intra-observer repeatability [all parameters evaluated had low CoV (<0.71%) and high ICC (>0.88)]. Inter-observer reproducibility was also excellent, with high ICC (>0.95) and low CoV (<0.52%). The 95% LoA between the new biometer and OA-2000 were insignificant for most of the parameters evaluated, especially for AL. However, the measurement agreement was moderate for CCT. CONCLUSIONS: Intra-observer repeatability and inter-observer reproducibility were excellent for all parameters evaluated using the new optical biometer based on Scheimpflug imaging and PCI. There was a high agreement between the two devices and hence could be clinically interchangeable for the measurement of most ocular parameters.

Hepatitis C virus core protein-induced miR-93-5p up-regulation inhibits interferon signaling pathway by targeting IFNAR1.

AIM: To investigate the mechanism by which hepatitis C virus (HCV) core protein-induced miR-93-5p up-regulation regulates the interferon (IFN) signaling pathway. METHODS: HCV-1b core protein was exogenously expressed in Huh7 cells using pcDNA3.1 (+) vector. The expression of miR-93-5p and interferon receptor 1 (IFNAR1) was measured using quantitative reverse transcription-polymerase chain reaction and Western blot. The protein expression and phosphorylation level of STAT1 were evaluated by Western blot. The overexpression and silencing of miR-93-5p and IFNAR1 were performed using miR-93-5p agomir and antagomir, and pcDNA3.1-IFNAR1 and IFNAR1 siRNA, respectively. Luciferase assay was used to identify whether IFNAR1 is a target of miR-93-5p. Cellular experiments were also conducted. RESULTS: Serum miR-93-5p level was increased in patients with HCV-1b infection and decreased to normal level after HCV-1b clearance, but persistently increased in those with pegylated interferon-α resistance, compared with healthy subjects. Serum miR-93-5p expression had an AUC value of 0.8359 in distinguishing patients with pegylated interferon-α resistance from those with pegylated interferon-α sensitivity. HCV-1b core protein increased miR-93-5p expression and induced inactivation of the IFN signaling pathway in Huh7 cells. Furthermore, IFNAR1 was identified as a direct target of miR-93-5p, and IFNAR1 restore could rescue miR-93-5p-reduced STAT1 phosphorylation, suggesting that the miR-93-5p-IFNAR1 axis regulates the IFN signaling pathway. CONCLUSION: HCV-1b core protein-induced miR-93-5p up-regulation inhibits the IFN signaling pathway by directly targeting IFNAR1, and the miR-93-5p-IFNAR1 axis regulates STAT1 phosphorylation. This axis may be a potential therapeutic target for HCV-1b infection.

Metabolism studies on prim-O-glucosylcimifugin and cimifugin in human liver microsomes by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry.

Prim-O-glucosylcimifugin (PGCN) and cimifugin (CN) are major constituents of Radix Saposhnikoviae that have antipyretic, analgesic and anti-inflammatory pharmacological activities. However, there were few reports with respect to the metabolism of PGCN and CN in vitro. In this paper, we describe a strategy using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) for fast analysis of the metabolic profile of PGCN and CN in human liver microsomes. In total, five phase I metabolites of PGCN, seven phase I metabolites and two phase II metabolites of CN were identified in the incubation of human liver microsomes. The results revealed that the main phase I metabolic pathways of PGCN were hydroxylation and hydrolysis reactions. The phase I metabolic pathways of CN were found to be hydroxylation, demethylation and dehydrogenation. Meanwhile, the results indicated that O-glucuronidation was the major metabolic pathway of CN in phase II metabolism. The specific UDP-glucuronosyltransferase (UGT) enzymes responsible for CN glucuronidation metabolites were identified using recombinant UGT enzymes. The results indicated that UGT1A1, UGT1A9, UGT2B4 and UGT2B7 might play major roles in the glucuronidation of CN. Overall, this study may be useful for the investigation of metabolic mechanism of PGCN and CN, and it can provide reference and evidence for further pharmacodynamic experiments. Copyright © 2016 John Wiley & Sons, Ltd.

Simultaneous Determination of Five Components in Aster tataricus by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry.

A novel quantitative method using high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry was developed for simultaneous determination of the important active constituents including shionone, luteolin, quercetin, kaempferol and ferulic acid in Aster tataricus from different habitats. The separation was performed on a C18 column with acidified aqueous acetonitrile gradients. Quantification of the analytes was achieved by the use of a hybrid quadrupole spectrometer. Multiple reaction monitoring scanning was employed with positive and negative modes at the same time in a single run. The validated results of the method indicated that the method was simple, rapid, specific and reliable. The results demonstrated that the quantitative difference in content of five active compounds was useful not only for chemotaxonomy of numerous samples from different sources but also for the standardization and differentiation of several similar samples. It was the first time to report a UPLC-ESI-MS-MS method for determination of five components in A. tataricus extract. Simultaneous quantification of bioactive components by UPLC-ESI-MS could be a well-acceptable strategy to control the quality of A. tataricus extract comprehensively.

Tentative identification of new metabolites of cnidilin by liquid chromatography-mass spectrometry.

Cnidilin is one of the major bioactive constituents of Radix Angelicae dahuricae. The present study was designed to characterize and interpret the structures of metabolites in rats after oral administration of cnidilin at a dose of 48mg/kg body weight. Metabolite identification was accomplished using a predictive multiple reaction monitoring-information-dependent acquisition-enhanced product ion (pMRM-IDA-EPI) scan and precursor ion scan-information-dependent acquisition-enhanced product ion (PREC-IDA-EPI) scan in positive ion mode. Comparing the changes in protonated molecular masses, MS/MS spectra and retention times with the parent drug, 18 metabolites were identified. The result shows that the metabolic pathways contain deisopentenyl, combination with glucose, hydroxylation, oxidation, demethylation and addition reaction. The study identified 18 metabolites, analyzed and summarized the fragmentation regularities of mass spectra of 8-methoxy-5-hydroxy psoralen. The study provides a new pathway to discovery new compounds, new fragmentation regularities and the mode of metabolites.

LC-MS/MS determination and pharmacokinetic study of seven flavonoids in rat plasma after oral administration of Cirsium japonicum DC. extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Cirsium japonicum DC., a Traditional Chinese Medicine, has the curative effect of antihemorrhagic and antitumor. Pharmacological studies prove that the curative effect may relate to the flavonoids. A simple and rapid resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was first developed and validated for the quantification of seven flavonoids including pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and apigenin in rat plasma after oral administration of Cirsium japonicum DC. extract. MATERIALS AND METHODS: The chromatographic separation was achieved on a C18 column with gradient elution by using a mixture of 0.1% formic acid aqueous solution and methanol as the mobile phase at a flow rate of 0.8mL/min. A tandem mass spectrometric detection was conducted using multiple-reaction monitoring (MRM) via an electrospray ionization (ESI) source in positive and negative ionization mode simultaneously. Samples were pre-treated by a single-step protein precipitation with methanol, and sulfamethoxazole was used as internal standard (IS). RESULTS: The optimized mass transition ion-pairs (m/z) for quantization were 623.4/315.2 for pectolinarin, 593.3/285.1 for linarin, 315.3/300.2 for pectolinarigenin, 301.2/286.2 for hispidulin, 301.2/258.2 for diosmetin, 283.0/267.9 for acacetin, 269.0/117.0 for apigenin and 252.2/155.8 for IS. After oral administration of 6mL/kg Cirsium japonicum DC. extract in rats, the maximum plasma concentration (Cmax) of pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and apigenin were 876.77±97.34ng/mL, 86.79±1.70ng/mL, 6.13±0.12ng/mL, 32.85±2.50ng/mL, 37.2±2.04ng/mL, 19.02±1.29ng/mL and 148.26±20.63ng/mL, respectively. The time to reach the maximum plasma concentration (Tmax) was 5min for pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and 360min for apigenin. The intra-day and inter-day precisions (RSD%) for seven compounds were less than 13.16% and 7.77% and the accuracy (RE%) range from -7.92% to 14.77%. CONCLUSIONS: This is the first research on the pharmacokinetic study of bioactive components in rat plasma after oral administration of Cirsium japonicum DC. extract. The results provided a meaningful basis for better understanding the absorption mechanism of Cirsium japonicum DC. and evaluating the clinical application of this herb medicine.