Effects of Designed Herbal Formula on Growth Performance, Blood Indices, Organ Traits, and Cecum Microbiology in Broilers.

The objective of this study was to investigate the effect of the designed herbal formula (DHF) on growth performance, blood indices, organ traits, and cecum microbiology in broilers. A total of 96 male broilers of 1 d were selected and randomly assigned to two groups with six replicates of eight broilers each. The control (CON) and the basal diet containing 1.0% DHF (Astragali radix, Atractylodes macrocephala Koidz., Isatis tinctoria Linnaeus, and Citri reticulatae pericarpium, 2:1:1:2) were fed separately. The experiment was conducted for 35 days. The results showed that the DHF diet increased body weight and decreased the feed conversion ratio (FCR) (p < 0.05). At 21 days, the spleen, thymus, lymphocytes, and thrombocytes were increased (p < 0.05), and pancreas, duodenum, heterophils, and mean corpuscular hemoglobin (MCH) were decreased (p < 0.05). At 35 days, the heart, pancreas, white blood cell, heterophils, hemoglobin, MCH and mean corpuscular hemoglobin concentration (MCHC) were decreased, while lymphocytes and middle cells were increased (p < 0.05). The results of microbial diversity analysis showed that the DHF diet decreased the microbial diversity of the cecum. Firmicutes and Bacteroidetes were the dominant phyla, where the DHF diet increased the relative abundances of Bacteroides uniformis, Bacteroides vulgatus, and Faecalibacterium prausnitzii, and then decreased the relative abundance of Shigella sonnei. In conclusion, DHF played a positive role in improving the growth performance, immune performance, and relative abundance of Bacteroides uniformis, Bacteroides vulgatus, and Faecalibacterium prausnitzii in cecum microbiology in broilers, and has the potential to be used as a novel feed additive.

Characterization of the Endophytic Bacillus subtilis KRS015 Strain for Its Biocontrol Efficacy Against Verticillium dahliae.

Endophytes play important roles in promoting plant growth and controlling plant diseases. Verticillium wilt is a vascular wilt disease caused by Verticillium dahliae, a widely distributed soilborne pathogen that causes significant economic losses on cotton each year. In this study, an endophyte KRS015, isolated from the seed of the Verticillium wilt-resistant Gossypium hirsutum 'Zhongzhimian No. 2', was identified as Bacillus subtilis by morphological, phylogenetic, physiological, and biochemical analyses. The volatile organic compounds (VOCs) produced by KRS015 or its cell-free fermentation extract had significant antagonistic effects on various pathogenic fungi, including V. dahliae. KRS015 reduced Verticillium wilt index and colonization of V. dahliae in treated cotton seedlings significantly; the disease reduction rate was ∼62%. KRS015 also promoted plant growth, potentially mediated by the growth-related cotton genes GhACL5 and GhCPD-3. The cell-free fermentation extract of KRS015 triggered a hypersensitivity response, including reactive oxygen species (ROS) and expression of resistance-related plant genes. VOCs from KRS015 also inhibited germination of conidia and the mycelial growth of V. dahliae, and were mediated by growth and development-related genes such as VdHapX, VdMcm1, Vdpf, and Vel1. These results suggest that KRS015 is a potential agent for controlling Verticillium wilt and promoting growth of cotton.

Sleep deprivation affects sex hormones secretion by regulating the expression of the circadian clock gene in the hypothalamus and pituitary via the PI3K/Akt signaling pathway in pregnant rats.

Sleep deprivation (SD) has many deleterious health effects and occurs in more than 70% of pregnant women. However, the changes in sex hormones and relevant mechanisms after SD have not been well clarified. The aim of the present study was to explore the effects of SD on the secretion of sex hormones and the underlying mechanisms. Twelve pregnant Wistar rats were divided into control (CON, n = 6) and SD (n = 6) groups. Pregnant rats in the SD group were deprived of sleep for 18 h, and allowed free rest for 6 h, and then the above procedures were repeated until delivery. The CON group lived in a 12 h light/dark light cycle environment. Estradiol (E2) and progesterone (P4) levels were detected by enzyme-linked immunosorbent assay (ELISA), and the expression of circadian clock genes, Bmal1, Clock and Per2, in hypothalamus and pituitary gland tissues were evaluated by immunohistochemistry (IHC) and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The PI3K and Akt phosphorylation levels in the hypothalamic and pituitary tissues were determined by Western blot. The results showed that, compared with the CON group, the SD group exhibited significantly reduced serum E2 and P4 levels, down-regulated Bmal1, Clock and Per2 expression, as well as decreased phosphorylation levels of PI3K and Akt. But there was no significant difference of the total PI3K and Akt protein expression levels between the two groups. These results suggest that SD might affect the expression of the circadian clock genes in the hypothalamus and pituitary via PI3K/Akt pathway, and subsequently regulate the secretion of sex hormones in the pregnant rats, which hints the important roles of SD-induced changes of serum sex hormone levels in the pregnant rats.

Cold atmospheric plasmas target breast cancer stemness via modulating AQP3-19Y mediated AQP3-5K and FOXO1 K48-ubiquitination.

Cold atmospheric plasma (CAP) is selective against many cancers with little side effect, yet its molecular mechanism remains unclear. Through whole transcriptome sequencing followed by assays in vitro, in vivo and using clinical samples, we propose CAP as a promising onco-therapy targeting cancer stemness via the AQP3/FOXO1 axis. CAP-generated reactive species penetrated cells via AQP3 and suppressed RPS6KA3, a shared kinase of AQP3 and FOXO1. Reduced AQP3-19Y phosphorylation suppressed SCAF11-mediated AQP3-5K K48-ubiquitination that led to sabotaged FOXO1 stability. Inhibited FOXO1 phosphorylation retarded its regulatory activities in maintaining cancer stemness including ALDH1 and IL6. Enhanced anti-cancer efficacy was observed through combining CAP with Atorvastatin in vitro and in vivo. We propose CAP as a 'selective' onco-therapeutic against cancer stemness, with the AQP3/FOXO1 axis being one molecular mechanism. We report SCAF11 as an E3 ubiquitin ligase of both AQP3 and FOXO1, identify AQP3-5K as an AQP3 K48-ubiquitination site, and emphasize the essential role of AQP3-19Y in this process. We reposition Atorvastatin into the onco-therapeutic portfolio by synergizing it with CAP towards enhanced efficacy. We anticipate the efficacy of CAP in targeting malignancies of high stemness alone or as an adjuvant therapy towards the hope of ultimate cancer cure.

Proteomics Analysis of Serum Proteins in Gestational Diabetes.

OBJECTIVE: The purpose of this study was to screen serum proteins for biomarkers of gestational diabetes mellitus (GDM) and to investigate its pathogenesis by analyzing the differences in serum proteomics between pregnant women with GDM and healthy pregnant women. METHODS: Patients who were admitted to the First Affiliated Hospital of Fujian Medical University from June 2019 to January 2020 were included. According to the medical history and the results of the 75 g oral glucose tolerance test (OGTT), they were divided into the normal pregnant women group and GDM pregnant women group. The serum of two groups of patients was collected. High performance liquid chromatography-mass spectrometry was used to identify differentially expressed serum proteins between pregnant women with GDM and healthy pregnant women, and bioinformatics analysis was then performed on the identified proteins. RESULTS: A total of 1152 quantifiable proteins were detected; among them, 15 were upregulated in serum of GDM pregnant women, while 26 were downregulated. The subsequent parallel reaction monitoring (PRM) assay validated the expression levels of 12 out of 41 differentially expressed proteins. Moreover, bioinformatics analysis revealed that the differentially expressed proteins are involved in multiple biological processes and signaling pathways related to the lipid metabolism, glycan degradation, immune response, and platelet aggregation. CONCLUSIONS: This study identified 41 serum proteins with differential expression between pregnant women with GDM and healthy pregnant women, providing new candidate molecules for elucidating GDM pathogenesis and screening therapeutic targets.

Rhizosphere Microbiomes of Potato Cultivated under Bacillus subtilis Treatment Influence the Quality of Potato Tubers.

Plants serve as a niche for the growth and proliferation of a diversity of microorganisms. Soil microorganisms, which closely interact with plants, are increasingly being recognized as factors important to plant health. In this study, we explored the use of high-throughput DNA sequencing of the fungal ITS and bacterial 16S for characterization of the fungal and bacterial microbiomes following biocontrol treatment (DT) with Bacillus subtilis strain Bv17 relative to treatments without biocontrol (DC) during the potato growth cycle at three time points. A total of 5631 operational taxonomic units (OTUs) were identified from the 16S data, and 2236 OTUs were identified from the ITS data. The number of bacterial and fungal OTU in DT was higher than in DC and gradually increased during potato growth. In addition, indices such as Ace, Chao, Shannon, and Simpson were higher in DT than in DC, indicating greater richness and community diversity in soil following the biocontrol treatment. Additionally, the potato tuber yields improved without a measurable change in the bacterial communities following the B. subtilis strain Bv17 treatment. These results suggest that soil microbial communities in the rhizosphere are differentially affected by the biocontrol treatment while improving potato yield, providing a strong basis for biocontrol utilization in crop production.

Cantharidin-loaded biomimetic MOF nanoparticle cascade to enhance the Fenton reaction based on amplified photothermal therapy.

The treatment efficiency of the Fenton reaction is expected to be greatly restricted due to problems such as inefficient delivery of Fenton catalysis, limited H2O2 concentration and uneven tumour tissue. Accurate photothermal therapy (PTT) could improve the efficiency of Fenton catalysis to some extent by raising the temperature. However, the heat shock response (HSR) of tumour cells caused by PTT and Fenton reaction would attenuate the treatment effect. In this study, we developed an iron ions-mediated Fenton reaction combined with a PTT treatment platform based on a metal-organic framework, i.e., PPy-CTD@MIL-100@MPCM nanoparticles (PCMM NPs), and further explored the inhibitory effect of PCMM NPs on the heat shock response (HSR). PCMM NPs could accumulate in tumour tissue via the coated macrophage cell membranes (MPCMs) to target inflammatory tissues. The photothermal effect of polypyrrole (PPy) accelerated the release of cantharidin (CTD) and iron ions loaded in the PCMM NPs. CTD, as an HSR inhibitor, could inhibit this response of tumour cells and improve the effect of PTT. Meanwhile, the heat generated during the PTT process could improve the efficiency of the Fenton reaction. This study suggested that PCMM NPs could serve as a combined treatment platform to enhance the Fenton reaction based on amplified photothermal therapy.

Cu/Zn superoxide dismutase (VdSOD1) mediates reactive oxygen species detoxification and modulates virulence in Verticillium dahliae.

The accumulation of reactive oxygen species (ROS) is a widespread defence mechanism in higher plants against pathogen attack and sometimes is the cause of cell death that facilitates attack by necrotrophic pathogens. Plant pathogens use superoxide dismutase (SOD) to scavenge ROS derived from their own metabolism or generated from host defence. The significance and roles of SODs in the vascular plant pathogen Verticillium dahliae are unclear. Our previous study showed a significant upregulation of Cu/Zn-SOD1 (VdSOD1) in cotton tissues following V. dahliae infection, suggesting that it may play a role in pathogen virulence. Here, we constructed VdSOD1 deletion mutants (ΔSOD1) and investigated its function in scavenging ROS and promoting pathogen virulence. ΔSOD1 had normal growth and conidiation but exhibited significantly higher sensitivity to the intracellular ROS generator menadione. Despite lacking a signal peptide, assays in vitro by western blot and in vivo by confocal microscopy revealed that secretion of VdSOD1 is dependent on the Golgi reassembly stacking protein (VdGRASP). Both menadione-treated ΔSOD1 and cotton roots infected with ΔSOD1 accumulated more O2- and less H2 O2 than with the wildtype strain. The absence of a functioning VdSOD1 significantly reduced symptom severity and pathogen colonization in both cotton and Nicotiana benthamiana. VdSOD1 is nonessential for growth or viability of V. dahliae, but is involved in the detoxification of both intracellular ROS and host-generated extracellular ROS, and contributes significantly to virulence in V. dahliae.

Potato resistant starch inhibits diet-induced obesity by modifying the composition of intestinal microbiota and their metabolites in obese mice.

Potato resistant starch type 3 (PRS) is helpful for weight-loss. To investigate the regulatory effects of PRS on high-fat diet (HFD)-induced obesity, different doses of PRS (5%, 15% and 25%) were fed to mice for 12 weeks. Metabolic syndrome related to obesity, intestinal microbiota composition and its metabolites as well as the relationship among them were studied. Results showed that PRS could regulate HFD-induced metabolic syndrome in a dose dependent manner; promote the proliferation of intestinal cells and expression of tight junction proteins, such as Occludin and zonula occludens (ZO)-1; reduce the Firmicutes/Bacteroidetes (F/B) rate; regulate the relative abundance of intestinal microbiota, such as Bifidobacterium, Ruminococcus, Bacteroides and Coprococcus; and promote the production of microbial metabolites, such as propionic acid and acetic acid. Besides, the alteration in the intestinal microbiota composition and metabolites were significantly correlated. It could be concluded that propionic acid and acetic acid were the two dominant metabolites of Bifidobacterium, Ruminococcus, Bacteroides, and Coprococcus, which contributed to the anti-obesity potential of PRS, metabolic syndrome alleviation, and intestinal barrier dysfunction.

Screening of pesticide residues in Traditional Chinese Medicines using modified QuEChERS sample preparation procedure and LC-MS/MS analysis.

A robust and high-throughput method was developed for the determination of 108 pesticide residues in Traditional Chinese Medicines (TCMs) simultaneously using a combination of UHPLC-MS/MS analysis and the modified QuEChERS method. Extraction was carried out in acetonitrile containing 0.75% (v/v) acetic acid with ultrasonication for 15 min; MgSO4 and C18 were used as the dispersive-solid phase extraction sorbents. The method exhibited good linearity (r2 > 0.9901), in addition to good selectivity, precision and repeatability. More than 92% of pesticides exhibited high rates or recovery in the 70-120% range. This method showed high sensitivity, with Limits of Quantitation in the 0.01-20 ng/mL range in Cortex Moutan, and 0.01-50 ng/mL in the other TCMs. The method was employed for the analysis of 39 real samples from different habitats, and pesticides were detected in 92.3% of the samples, with 26 pesticides being detected in these three TCMs. More than four pesticides were detected in a third of the samples. Among them, tebuconazole was detected in all the three TCMs with 0.22-22.02 µg/kg concentration, which was lower than the provisions in GB 2763-2019 (50 µg/kg). In addition, the paclobutrazol detection rate in Ophiopogon japonicus was 100%, and the detected concentrations of 9 samples exceeded the Maximum Residue Levels defined for vegetables (50 µg/kg). Considering there are no regulations that govern the limits of pesticide residues in the three TCMs in China, we recommend the acceleration of efforts to introduce appropriate regulations.

Comparative Study of the Nutritional Properties of 67 Potato Cultivars (Solanum tuberosum L.) Grown in China Using the Nutrient-Rich Foods (NRF11.3) Index.

The overall nutritional properties of tubers from 67 potato cultivars were systematically evaluated in this study by adopting the Nutrient-Rich Foods (NRF11.3) Index Model. The macronutrients including dry matter, crude protein, total dietary fiber, and starch contents were found to be in the range of 14.8-30.5 g/100 g fresh weight, 5.71-12.0, 1.99-3.39, and 56.0-75.5 g/100 g dry weight, respectively. Additionally, the amounts of vitamin C, K and Fe were 22.6-86.6, 1457-3111, and 1.40-5.06 mg/100 g dry weight, respectively. The NRF11.3 index model has a score of 66.4-102 per 100 kcal for male and 70.8-107 per 100 kcal for female over 18 years old. This model was utilized to determine the macrocomponents and micronutrients of diverse potato cultivars and aid in comprehensive nutritional study on potato as a desirable raw material for staple food processing to human nutrition and daily intake.

Recent mycotoxin survey data and advanced mycotoxin detection techniques reported from China: a review.

Mycotoxin contamination in agro-food systems has been a serious concern over the last few decades in China, where the Ministry of Health has set maximum limits for mycotoxins in different agro-products. Overall survey data show that aflatoxin contamination in infant cereals, edible oils, raw milk, ginger and its related products are far below Chinese regulatory limits. The absence of aflatoxin M1 contamination in infant milk powders indicates a high standard of control. Aflatoxins in liquorice roots and lotus seeds have been reported for the first time. For deoxynivalenol, high levels were found in wheat grown in the Yangtze Delta region, which is more prone to rainfall, supporting Fusarium infection. The emerging mycotoxins beauvericins and enniatins have been reported in the medicinal herbs in China. Ochratoxin A in wine was below the European Union regulatory limits, but fumonisins in maize need to be monitored and future regulatory control considered. Overall from all the survey data analysed in this review, it can be concluded that 92% of the samples analysed had mycotoxin levels below the Chinese regulatory limits. In terms of detection techniques in recent years, immuno-based assays have been developed largely due to their excellent sensitivity and ease of use. Assays targeting multiple mycotoxins like aflatoxins, ochratoxin A, zearalenone and deoxynivalenol have been reported using microarrays and suspension arrays targeting in particular maize, rice and peanuts. Aptamer-based assays against ochratoxin A and aflatoxins B1 and B2 have been developed involving fluorescence detection; and surface plasmon resonance immunosensors have been developed targeting wine, maize, wheat, wild rye, hay and peanut oil with high sensitivity (> 0.025 ng l(-1)). Commercialisation of these technologies is much needed for wider usage in the coming years.