Effects of Active Dry Yeast Supplementation in In Vitro and In Vivo Nutrient Digestibility, Rumen Fermentation, and Bacterial Community.

This study assessed the impact of active dry yeast (ADY) on nutrient digestibility and rumen fermentation, using both in vitro and in vivo experiments with lambs. In vitro, ADYs were incubated with rumen fluid and a substrate mixture to assess gas production, pH, volatile fatty acid (VFA) profiles, and lactate concentration. In vivo, Hu lambs were randomly assigned to five dietary treatments: a control group and four groups receiving one of two dosages of either Vistacell or Procreatin7. Growth performance, nutrient digestibility, rumen fermentation parameters, and bacterial community composition were measured. Pro enhanced the propionate molar proportion while it decreased the n-butyrate molar proportion. Vis reduced the lactate concentration in vitro. In the in vivo experiment, Vis increased the propionate molar proportion and the Succinivibrionaceae_UCG-001 abundance while it decreased the n-butyrate molar proportion and the Lachnospiraceae_ND3007 abundance. Additionally, Vis showed a greater impact on improving the NDF digestibility and total VFA concentration in vivo compared to Pro. Overall, the effects of ADYs on rumen fermentation were found to vary depending on the specific ADY used, with Vis being the most suitable for lamb growth. It was observed that Vis promoted propionate fermentation and Succinivibrionaceae_UCG-001 abundance at the expense of reduced n-butyrate fermentation and Lachnospiraceae_ND3007 abundance. Importantly, differences were noted between the outcomes of the in vitro and in vivo experiments concerning the effects of ADYs on rumen fermentation, highlighting the need for caution when generalizing batch culture results to the in vivo effects of ADYs.

MiR-122-5p regulates erastin-induced ferroptosis via CS in nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) is a tumor that occurs in the nasopharynx. Although advances in detection and treatment have improved the prognosis of NPC the treatment of advanced NPC remains challenging. Here, we explored the effect of microRNA (miR)-122-5p on erastin-induced ferroptosis in NPC cells and the role of ferroptosis in the development of NPC. The effect of miR-122-5p silencing and overexpression and the effect of citrate synthase on erastin-induced lipid peroxidation in NPC cells was analyzed by measuring the amounts of malondialdehyde, Fe2+, glutathione, and reactive oxygen species and the morphological alterations of mitochondria. The malignant biological behavior of NPC cells was examined by cell counting kit-8, EDU, colony formation, Transwell, and wound healing assays. The effects of miR-122-5p on cell proliferation and migration associated with ferroptosis were examined in vivo in a mouse model of NPC generated by subcutaneous injection of NPC cells. We found that erastin induced ferroptosis in NPC cells. miR-122-5p overexpression inhibited CS, thereby promoting erastin-induced ferroptosis in NPC cells and decreasing NPC cell proliferation, migration, and invasion.

Enhanced catalytic ozonation performance by CuOx nanoclusters/TiO2 nanotube and an insight into the catalytic mechanism.

Highly reactive nanoclusters of metal oxides are extremely difficult to be synthesized due to their thermodynamic instability. For the first time, CuOx nanoclusters supported on anatase TiO2 nanotubes (NT) with many defects as anchoring sites were successfully prepared. Although the copper loading reached as high as 2.5 %, the size of CuOx nanoclusters in the sample of 2.5 %CuOx/NT were mainly around 1.0 nm. The aggregation of copper species during the calcination process was undoubtedly hampered by the anchoring effects of the abundant defects in NT support. Due to the highly exposed undercoordinated atoms of CuOx nanoclusters, the mixed valences of copper, and the strong interface interaction between CuOx nanoclusters and NT support, 2.5 %CuOx/NT-catalyzed ozonation showed the highest pseudo-first-order reaction rate constant of 8.5 × 10-2 min-1, 2.2 and 4.0 times that of NT-catalyzed ozonation and ozonation alone, respectively. Finally, the catalytic mechanism was revealed by both experiments and density functional theory calculations (DFT). The results demonstrated that the undercoordinated Cu in CuOx/NT could highly promote the adsorption of ozone with a high adsorption energy of -125.16 eV and the adsorbed ozone was activated immediately, tending to dissociate into a O2 molecule and a surface O atom. Thus, abundant reactive oxygen species, e.g., hydroxyl radical (·OH), superoxide radical (·O2-) and singlet oxygen (1O2), could be generated via chain reactions. Especially, ·OH mainly contributed to the removal of ibuprofen pollutants. This work sheds a light on the design and preparation of highly reactive nanoclusters of metal oxide catalysts for catalytic ozonation of refractory organic pollutants.

Identification of key genes and immune infiltration based on weighted gene co-expression network analysis in vestibular schwannoma.

Vestibular schwannomas are the most common tumors of the cerebellopontine angle, but their pathogenesis is still unclear. This study aimed to explore the molecular mechanisms and potential therapeutic target biomarkers in vestibular schwannoma. Two datasets (GSE141801 and GSE54934) were downloaded from the Gene Expression Omnibus database. Weighted gene coexpression network analysis was performed to find the key modules associated with vestibular schwannoma (VS). Functional enrichment analysis was applied to evaluate the gene enrichment signaling pathway in key modules. Protein-protein interaction networks in key modules were constructed using the STRING website. Hub genes were identified by intersecting candidate hub genes in protein-protein interaction network and candidate hub genes in key modules. Single-sample gene set enrichment analysis was utilized to quantify the abundance of tumor-infiltrating immune cells in VSs and normal control nerves. A Random forest classifier was developed based on hub genes identified in this study and validated on an independent dataset (GSE108524). Results of immune cell infiltration were also validated on GSE108524 by gene set enrichment analysis. Eight genes from coexpression modules were identified as hub genes, that is, CCND1, CAV1, GLI1, SOX9, LY86, TLR3, TREM2, and C3AR1, which might be potential therapeutic targets for VS. We also found that there were distinct differences in the infiltration levels of immune cells between VSs and normal control nerves. Overall, our findings may be useful for investigating the mechanisms underlying VS and provide noteworthy directions for future research.

MiR-223-3p regulates the eosinophil degranulation and enhances the inflammation in allergic rhinitis by targeting FBXW7.

OBJECTIVES: MiR-223-3p is a multifunctional microRNA regulated by multiple transcription factors and plays a critical role in inflammation. This paper was designed to investigate the regulatory role and mechanism of miR-223-3p in eosinophils degranulation and allergic rhinitis (AR) inflammation. METHODS: OVA sensitized AR mouse model and EOL-1 cells model were established. RT-qPCR and FISH were performed to detect the miR-223-3p expression. ELISA and WB were utilized to evaluate mRNA and protein expression. HE staining and transmission electron microscopy were applied to observe the morphological changes in nasal mucosa. Flow cytometry and immunofluorescence staining were performed to measure the proportion of eosinophils and eosinophilic major basic protein expression. The targeting relationship between miR-223-3p and FBXW7 was verified by bioinformatic analysis and dual-luciferase reporter gene assay. The expression of FBXW7 was detected by immunohistochemistry. RESULTS: The level of miR-223-3p in nasal mucosa was significantly up-regulated in AR group. The expression of miR-223-3p, ECP, MBP, and EPO were increased in EOL-1 cells, further increasing the miR-223-3p level could promote the ECP and EPO mRNA expression. Upregulation of miR-223-3p increased eosinophils granule protein expression, aggravated mucosal destruction and enhanced AR inflammation. Luciferase assay verified miR-223-3p directly target the 3'-UTR of FBXW7. In vitro, overexpression of FBXW7 could reverse the increase in MBP expression caused by the up-regulation of miR-223-3p. In vivo, knockdown of FBXW7 could reverse the down-regulation in granule protein level caused by the down-regulation of miR-223-3p, thereby aggravating AR inflammation. CONCLUSION: Collected evidence elucidated that miR-223-3p could regulate the eosinophil degranulation and enhances the inflammation in AR by targeting FBXW7. The miR-223-3p/FBXW7 axis may provide a novel approach for AR treatment.

The role of microRNAs in ferroptosis.

Ferroptosis is a newly discovered type of programmed cell death, which is closely related to the imbalance of iron metabolism and oxidative stress. Ferroptosis has become an important research topic in the fields of cardiomyopathy, tumors, neuronal injury disorders, and ischemia perfusion disorders. As an important part of non-coding RNA, microRNAs regulate various metabolic pathways in the human body at the post-transcriptional level and play a crucial role in the occurrence and development of many diseases. The present review introduces the mechanisms of ferroptosis and describes the relevant pathways by which microRNAs affect cardiomyopathy, tumors, neuronal injury disorders and ischemia perfusion disorders through regulating ferroptosis. In addition, it provides important insights into ferroptosis-related microRNAs, aiming to uncover new methods for treatment of the above diseases, and discusses new ideas for the implementation of possible microRNA-based ferroptosis-targeted therapies in the future.

The association between hearing threshold and urinary personal care and consumer product metabolites in middle-aged and elderly people from the USA.

Endocrine disruptors have been reported to be associated with hearing ability. However, the association between personal care and consumer product chemicals, known as commonly detected endocrine disruptors, and age-related hearing loss still remains unclear. This study aimed to examine the association between exposure to 7 personal care and consumer product chemicals and hearing thresholds in middle-aged and elderly people. A nationally representative cross-sectional study was performed. Eight hundred forty-five adults aged over 45 from the National Health and Nutrition Examination Survey (NHANES) were included in this study. Bayesian kernel machine regression (BKMR) and the k-medoid cluster analysis were used to evaluate the mixture effect of exposure to 7 chemicals on pure-tone average (PTA). Exposure to these chemicals was negatively associated with PTA. 2,5-Dichlorophenol had the greatest contribution to the mixture effect. The mixture effect was stronger in women, elderly people. Four pooled clusters were identified according to 7 chemicals exposures. Cluster 4 (high TCS exposure) showed a lower HFPTA (P = 0.00258) than cluster 3 (the lowest exposure cluster, as a reference). Our study provides evidence that exposure to personal care and consumer product chemicals might be inversely associated with PTA. More studies are needed to fully understand the association of exposure to these chemicals with hearing threshold.

The Inverted-U Relationship Between Dietary Inflammatory Potential and Hearing Loss Among Adults Aged 20 Years and Over in the United States: A Cross-Sectional Study.

PURPOSE: The purpose of this study was to examine the relationship between dietary inflammatory index and objective hearing loss (HL). MATERIALS AND METHODS: A cross-sectional analysis of a nationally representative sample of participants was performed based on data in National Health and Nutrition Examination Survey (NHANES) (2009-2016). HL was defined as pure tone averages >25 dB at 500, 1000, and 2000 Hz (low frequency); 3000, 4000, 6000, and 8000 Hz (high frequency) in either ear. The energy-adjusted dietary inflammatory index (E-DII) score was calculated for each participant based on two 24-h dietary recalls to assess diet-associated inflammation. Multivariable logistic regression was used to examine the linear relationship between HL and E-DII score or E-DII quartiles. Restricted cubic spline was applied to identify any non-linear associations of the E-DII score with hearing loss. Subgroup analyses were performed by age and gender to explore the moderating roles of these factors. Akaike's Information Criterion (AIC) values were used to select the better-fitted model among linear and non-linear models. RESULTS: An inverted-U shaped relationship with low-frequency hearing loss (LFHL) was identified for the E-DII score (P-nonlinear =0.023) after adjustment for potential confounders. But significant linear or nonlinear association between E-DII score and high-frequency hearing loss (HFHL) was not found. CONCLUSION: E-DII score had inverted-U relationship with LFHL. Both pro-inflammatory diet and anti-inflammatory diet seemed to be associated with a decreased risk of LFHL compared to diet that was neither pro-inflammatory diet nor anti-inflammatory diet.

Second Primary Malignancy in Patients with Hypopharyngeal Carcinoma: A SEER-Based Study.

BACKGROUND: A population-based analysis of the risk of secondary primary malignancy (SPM) in patients with hypopharyngeal carcinoma (HPC) has been lacking in the literature. Therefore, we conducted this study to determine the risk factors and assess the effects of SPM on the overall survival (OS) and cancer-specific survival (CSS) of patients with HPC. METHODS: Data on selected patients diagnosed with HPC from the Surveillance, Epidemiology and End Results (SEER) database between 1973 and 2015 were examined through logistic regression, Cox regression and nomogram methods. RESULTS: The overall risk of SPM in patients with HPC was higher than that in the general population (SIR: 2.77; P < 0.05). The specific-site, including the oral cavity, pharynx, digestive system, respiratory system and endocrine system, had a relatively higher risk of SPM. The overall risks of the subgroup of people 55-75 years of age and all subgroups of sex, race and latency were significantly elevated. In addition, patients with HPC were more likely to have been diagnosed in 2010-2015 (vs 2004-2009; P = 0.002), to be unmarried (vs married; P = 0.008), to have distant metastasis (vs no metastasis; P = 0.016) and to have had no surgery for the first tumor (vs surgery for the first tumor; P = 0.021), and these aspects were associated with a significantly elevated risk of developing SPM. SPM was independently associated with better OS and CSS. The OS and CSS in patients with HPC with SPM were better than those in patients without SPM (log rank P < 0.0001). The C indexes of the nomogram constructed with ten influencing factors including SPM were 0.681:0.699 for OS and 0.705:0.724 for CSS (training cohort:validation cohort). CONCLUSION: Although the overall risk of SPM in patients with HPC was elevated, SPM did not decrease the OS and CSS in patients with HPC. This finding is inconsistent with clinical observations and thus requires further research and exploration. It possibly because HPC might have a shorter survival time, or the follow-up time was not long enough.

MicroRNA-155-5p regulates the Th1/Th2 cytokines expression and the apoptosis of group 2 innate lymphoid cells via targeting TP53INP1 in allergic rhinitis.

As a key component of innate immunity, group 2 innate lymphoid cells (ILC2s) play a key role in Allergic rhinitis (AR). We previously demonstrated that both miR-155-5p and ILC2s are overexpressed in the nasal mucosa of AR patients, but the underlying mechanism remains unclear. At present study, we revealed that miR-155-5p was highly expressed in ILC2s of AR patients. Moreover, miR-155-5p promoted the secretion of Th2 cytokines of ILC2s, while inhibited the secretion of Th1 cytokines and the apoptosis of ILC2s. Meanwhile, the TP53INP1 expression was poorly expressed in ILC2s of AR patients. A dual luciferase reporter assay demonstrated that TP53INP1 was a direct target of miR-155-5p, and its expression was inversely associated with miR-155-5p in ILC2s. Furthermore, TP53INP1 inhibited the secretion of Th2 cytokines of ILC2s, while promoted the secretion of Th1 cytokines and the apoptosis of ILC2s. Notably, rescue experiments demonstrated that overexpression of TP53INP1 could partially reverse the effect of miR-155-5p on ILC2s. Taken together, these findings suggested that miR-155-5p aggravated the inflammatory response of AR dominated by ILC2s via targeting TP53INP1, which may aid in the development of novel therapeutic agents for AR.

Rapid and Visual Detection of Vibrio parahaemolyticus in Aquatic Foods Using blaCARB-17 Gene-Based Loop-Mediated Isothermal Amplification with Lateral Flow Dipstick (LAMP-LFD).

Vibrio parahaemolyticus is recognized as one of the most important foodborne pathogens responsible for gastroenteritis in humans. The blaCARB-17 gene is an intrinsic ß-lactamase gene and a novel species-specific genetic marker of V. parahaemolyticus. In this study, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) was developed targeting this blaCARB-17 gene. The specificity of LAMP-LFD was ascertained by detecting V. parahaemolyticus ATCC 17802 and seven other non-V. parahaemolyticus strains. Finally, the practicability of LAMP-LFD was confirmed by detection with V. parahaemolyticus-contaminated samples and natural food samples. The results showed that the optimized reaction parameters of LAMP are as follows: 2.4 mmol/l Mg2+, 0.96 mmol/l dNTPs, 4.8 U Bst DNA polymerase, and an 8:1 ratio of inner primer to outer primer, at 63°C for 40 min. The optimized reaction time of the LFD assay is 60 min. Cross-reactivity analysis with the seven non-V. parahaemolyticus strains showed that LAMP-LFD was exclusively specific for V. parahaemolyticus. The detection limit of LAMP-LFD for V. parahaemolyticus genomic DNA was 2.1 × 10-4 ng/µl, corresponding to 630 fg/reaction and displaying a sensitivity that is 100-fold higher than that of conventional PCR. LAMP-LFD in a spiking study revealed a detection limit of approximately 6 CFU/ml, which was similar with conventional PCR. The developed LAMP-LFD specifically identified the 10 V. parahaemolyticus isolates from 30 seafood samples, suggesting that this LAMP-LFD may be a suitable diagnostic method for detecting V. parahaemolyticus in aquatic foods.

Isolation, Molecular Characterization and Antibiotic Susceptibility Pattern of Vibrio parahaemolyticus from Aquatic Products in the Southern Fujian Coast, China.

Vibrio parahaemolyticus is a major gastroenteritis-causing pathogen in many Asian countries. Antimicrobial resistance in V. parahaemolyticus has been recognized as a critical threat to food safety. In this study, we determined the prevalence and incidence of antimicrobial resistance in V. parahaemolyticus in the southern Fujian coast, China. A total of 62 isolates were confirmed in retail aquatic products from June to October of 2018. The serotype O3:K6 strains, the virulence genes tdh and trh, antibiotic susceptibility and molecular typing were investigated. Then plasmid profiling analysis and curing experiment were performed for multidrug-resistant strains. The results showed that the total occurrence of V. parahaemolyticus was 31% out of 200 samples. Five strains (8.1%) out of 62 isolates were identified as the V. parahaemolyticus O3:K6 pandemic clone. A large majority of isolates exhibited higher resistance to penicillin (77.4%), oxacillin (71%), ampicillin (66.1%) and vancomycin (59.7%). Seventy-one percent (44/62) of the isolates exhibited multiple antimicrobial resistance. All 62 isolates were grouped into 7 clusters by randomly amplified polymorphic DNA, and most of the isolates (80.6%) were distributed within cluster A. Plasmids were detected in approximately 75% of the isolates, and seven different profiles were observed. Seventy-six percent (25/33) of the isolates carrying the plasmids were eliminated by 0.006% SDS incubated at 42°C, a sublethal condition. The occurrence of multidrug-resistant strains could be an indication of the excessive use of antibiotics in aquaculture farming. The rational use of antimicrobial agents and the surveillance of antibiotic administration may reduce the acquisition of resistance by microorganisms in aquatic ecosystems.

Micro-RNA-137 Inhibits Tau Hyperphosphorylation in Alzheimer's Disease and Targets the CACNA1C Gene in Transgenic Mice and Human Neuroblastoma SH-SY5Y Cells.

BACKGROUND Alzheimer's disease (AD) results in cognitive impairment. The calcium voltage-gated channel subunit alpha-1 C CACNA1C gene encodes an alpha-1 C subunit of L-type calcium channel (LTCC). The aim of this study was to investigate the role of micro-RNA-137 (miR-137) and the CACNA1C gene in APPswe/PS1ΔE9 (APP/PS1) double-transgenic AD mice and in human neuroblastoma SH-SY5Y cells. MATERIAL AND METHODS Six-month-old APP/PS1 double-transgenic AD mice (N=6) and age-matched normal C57BL/6 mice (N=6) underwent a Morris water maze (MWM) test, expression levels of amyloid-ß (Aß), LTCC, the CACNA1C gene, and miR-137 were measured in the rat hippocampus and cerebral cortex in both groups of mice. A luciferase assay was used to evaluate the effect of miR-137 on the expression of CACNA1C in SH-SY5Y human neuroblastoma SH-SY5Y cells. Western blotting was used to detect the CACNA1C, phosphorylated-tau (p-tau), and Aß proteins. RESULTS In APP/PS1 transgenic AD mice, spatial learning and memory was significantly reduced, levels of Aß1-40 and Aß1-42 were increased in the serum, hippocampus, and cerebral cortex, expression levels of miR-137 were reduced, expression of CACNA1C protein was increased in the hippocampus and cerebral cortex, compared with normal control mice. miR-137 regulated the expression of the CACNA1C gene. Increased expression levels of p-tau (Ser202, Ser396, and Ser404) induced by Aß1-42 were inhibited by miR-137 mimics in SH-SY5Y human neuroblastoma cells in vitro. CONCLUSIONS In a transgenic mouse model of AD, miR-137 and expression of the CACNA1C gene inhibited the hyperphosphorylation of tau protein.

Benign paroxysmal positional vertigo in irradiated nasopharyngeal carcinoma survivors.

Purpose. It has been assumed that postirradiated nasopharyngeal carcinoma (NPC) patients are prone to benign paroxysmal positional vertigo (BPPV). The purpose of this study was to better understand this clinical entity. Materials and Methods. From September 2003 to June 2011, we conducted a retrospective study of 11 irradiated NPC patients with BPPV in our institute. During the same period, 11 irradiated NPC patients without BPPV were randomly selected and enrolled as the control group. All medical records of these patients were evaluated. Results. The risk of BPPV rises significantly when the patient undergoes radiotherapy (RT) twice and the threshold radiation dose is >120 Gy (P = 0.027). The occurrence of postirradiated BPPV was significantly related to incidences of otitis media and sensorineural hearing loss (SNHL) (P = 0.011 and 0.009, resp.). All the patients responded well to repositioning maneuvers. Conclusion. A second course of RT, postirradiated otitis media, or SNHL is associated with the potential risk of radiation-induced BPPV. Repositioning maneuvers were safe and effective for relief of this disease.

[Extraction of organochlorine pesticides using ASE from wastewater irrigated agricultural soil].

The extraction efficiency of accelerated solvent extraction (ASE) was studied for organochlorine pesticide residuals in soil and compared with that of Soxhlet. The results showed that the efficiency of ASE was generally better than Soxhlet when used for DDT measurement, and equivalent to Soxhlet for BHCs. With ASE, organochlorine pesticide residues were detected in wastewater irrigated and non-wastewater irrigated soils from Tianjin. alpha-BHC, beta-BHC, delta-BHC, gamma-BHC, p,p'-DDE, p,p'-DDD, p,p'-DDT, o,p'-DDT ranged from 7.5 to 71.1 ng/g in wastewater irrigated vegetable and maize field and from 3.0 to 16.5 ng/g in wastewater irrigated paddy field. The eight pesticide residues from non-wastewater irrigated vegetable and maize field was in a range of 3.1-17.6 ng/g.