The objective of this study was to investigate the rumen degradation characteristics of grain amaranth hay (Amaranthus hypochondriacus) at four different growth stages. The aim of this study was to evaluate the nutritional value of grain amaranth hay at different growth stages by chemical composition, in vivo digestibility, and in situ degradability. Three Boer goats with permanent ruminal fistulas were selected in this study. Amaranthus hay at four different growth stages (squaring stage (SS), initial bloom stage (IS), full-bloom stage (FS) and mature stage (MS)) was crushed and placed into nylon bags. Each sample was set up with three replicates, and two parallel samples were set up in fistulas at each time point. The rumen degradation rates of dry matter (DM), crude protein (CP), neutral detergent fibre (NDF) and acid detergent fibre (ADF) were determined at 0, 6, 12, 24, 36, 48 and 72 h. The results were as follows: (1) The concentration of CP in SS was the highest and was significantly higher than that in other stages (p < 0.05), whereas the contents of NDF and ADF gradually increased with the extension of the growing period and reached a maximum in MS; (2)The degradation of CP in the rumen at 72 h of SS and IS was more than 80%. Compared with other stages, the effective degradability of CP was highest in SS (p < 0.05) and reached 87.05% at 72 h, and the degradation rate was the lowest in MS; and (3) The effective degradability of NDF in IS was the highest (p < 0.05) and reached 69.326% at 72 h. The effective degradability of ADF in MS was the highest (p < 0.05) and reached 65.728% at 72 h. The effective degradability of DM and CP in SS was the highest. In conclusion, among the four stages, IS was superior in chemical composition and rumen degradability characteristics.
To explore the genetic basis for a child featuring global developmental delay.DNA was extracted from peripheral blood sample taken from the patient and subjected to whole exome sequencing. Suspected variants were verified by Sanger sequencing of his family members.A heterozygous c.239T>C (p.Ile80Thr) variant of the GNB1 gene was detected in the proband, which was a verified to be de novo in origin.The heterozygous c.239T>C (p.Ile80Thr) variant of the GNB1 gene probably underlay the disease in this child.
Regulation of the expression of the prepro-TRH (ppTRH) gene by epidermal growth factor (EGF) was investigated. The i.p. injection of EGF significantly stimulated hypothalamic ppTRH messenger RNA levels in rats. To clarify whether this stimulatory effect of EGF could be exerted at the level of gene transcription, the 5'-flanking region (-1893/+127) of the mouse ppTRH gene fused to a luciferase reporter gene was transiently transfected into pituitary GH4C1 cells, and the effect of EGF on gene transcription was measured by a luciferase assay. EGF stimulated ppTRH gene promoter activity in a time- and dose-dependent manner. Deletion analysis revealed that two different regions of the promoter, between -254 and -218 [EGF response element-1 (EGFRE1)] and between -130 and -84 (EGFRE2) were required for full stimulation by EGF. The two EGFREs possessed putative binding sequences for the transcription factor Sp1, and they functioned cooperatively in heterologous promoters. Nuclear extracts from GH4C1 cells specifically bound those two EGFREs in gel retardation assays. Two protein-DNA complexes were found on EGFRE1, whereas four complexes were observed on EGFRE2. Although the binding of nuclear extracts to EGFRE1 was competed for by the consensus Sp1 binding sequence, the complexes on EGFRE1 were not supershifted by an Sp1 antibody. Formation of the slower migrating protein complex on EGFRE1 was prevented by EDTA, suggesting that one of the EGFRE1-binding proteins might be an Sp1-related zinc finger protein. Competition and supershift experiments demonstrated that the EGFRE2-binding protein showing that the slowest migration possessed a characteristic similar to that of Sp1. Selective mutations of the Sp1-binding site in EGFRE2 markedly diminished the EGF-induced stimulation. These results suggest that EGF may function as a positive regulator of ppTRH gene expression, and that the stimulatory effect may be mediated through a cooperative interaction between Sp1 or Sp1-related proteins and additional factors that bind to two separate DNA regions.
Excessive utilization of chemical fertilizers degrades the quality of medicinal plants and soil. Bio-organic fertilizers (BOFs) including microbial inoculants and microalgae have garnered considerable attention as potential substitutes for chemical fertilizer to enhance yield. In this study, a field experiment was conducted to investigate the effects of BOF partially substituting chemical fertilizer on the growth and quality of medicinal plant Polygala tenuifolia. The growth parameters, bioactive component contents, soil properties and composition of rhizosphere microorganisms were measured. The results indicated that substituting 40% of chemical fertilizer with microalgae showed the most pronounced growth-promoting effect, leading to a 29.30% increase in underground biomass and a 19.72% increase in 3,6'-disinapoylsucrose (DISS) content. Substituting 20% of chemical fertilizer with microalgae improved soil quality, significantly increasing soil organic matter content by 15.68% (p<0.05). Microalgae addition significantly affected the rhizosphere bacterial community composition of P. tenuifolia, reducing the relative abundance of Cladosporium by 33.33% and 57.93%, while increasing the relative abundance of Chloroflexi by 31.06% and 38.27%, under 20% and 40% chemical fertilizer reduction, respectively. The relative abundance of Chloroflexi positively correlated with both the underground biomass and DISS content (p<0.05), indicating that microalgae may stimulate Chloroflexi species associated with carbon cycling, thereby enhancing soil fertility, nutrient absorption, and ultimately leading to increased biomass accumulation and production of bioactive components in P. tenuifolia. In addition, there was no significant difference in underground growth and bioactive component contents between reduced chemical fertilizer dosage combined with solid microbial inoculant (SMI) and polyglutamic microbial inoculant (PMI), compared with 100% chemical fertilizer. Correlation analysis revealed that PMI could increase soil phosphorus availability through Streptomyces recruitment. In conclusion, our findings demonstrated that bio-organic fertilizers can partially substitute chemical fertilizer to improve soil properties and microorganisms, enhancing the growth and quality of P. tenuifolia. This provides a theoretical basis for increasing medicinal plant productivity under chemical fertilizer reduction.
After interpreting the reasons for protecting the ethnic minority′s cultural rights,this paper attempts to advocate a subject paradigm in people-oriented horizon in order to stress that the starting point and the end-result of protecting the ethnic minority′s cultural rights consists in human development and human happiness.Therefore,the disputes in the process of protection are defined and the legalized and systemized model of protecting the ethnic minority′s cultural rights is preliminarily constructed.
Presenting exogenous antigens on virus-like particles (VLPs) through 'plug-and-display' decoration strategies based on SpyTag/SpyCatcher isopeptide bonding have emerged as attractive technology for vaccine synthesis. However, whether the position of ligation site in VLPs will impose effects on immunogenicity and physiochemical properties of the synthetic vaccine remains rarely investigated. Here in the present work, the well-established hepatitis B core (HBc) protein was used as chassis to construct dual-antigen influenza nanovaccines, with the conserved epitope peptides derived from extracellular domain of matrix protein M2 (M2e) and hemagglutinin (HA) as target antigens. The M2e antigen was genetically fused to the HBc in the MIR region, together with the SpyTag peptide, which was fused either in the MIR region or at the N-terminal of the protein, so that a recombinant HA antigen (rHA) linked to SpyCatcher can be displayed on it, at two different localizations. Both synthetic nanovaccines showed ability in inducing strong M2e and rHA-specific antibodies and cellular immunogenicity; nevertheless, the one in which rHA was conjugated by N-terminal Tag ligation, was superior to another one synthesized by linking the rHA to MIR region SpyTagged-HBc in all aspects, including higher antigen-specific immunogenicity responses, lower anti-HBc carrier antibody, as well as better dispersion stability. Surface charge and hydrophobicity properties of the two synthetic nanovaccines were analyzed, results revealed that linking the rHA to MIR region SpyTagged-HBc lead to more significant and disadvantageous alteration in physiochemical properties of the HBc chassis.This study will expand our knowledge on 'plug-and-display' decoration strategies and provide helpful guidance for the rational design of HBc-VLPs based modular vaccines by using SpyTag/Catcher synthesis.
Increasing studies have shown that obesity is the primary cause of cardiovascular diseases, non-alcoholic fatty liver diseases, type 2 diabetes, and a variety of cancers. The dysfunction of gut microbiota was proved to result in obesity. Recent research indicated ANGPTL4 was a key regulator in lipid metabolism and a circulating medium for gut microbiota and fat deposition. The present study was conducted to investigate the alteration of gut microbiota and ANGPTL4 expression in the gastrointestinal tract of mice treated by the high-fat diet. Ten C57BL/6J mice were randomly allocated to two groups and fed with a high-fat diet (HFD) containing 60% fat or a normal-fat diet (Control) containing 10% fat. The segments of ileum and colon were collected for the determination of ANGPTL4 expression by RT-qPCR and immunohistochemical analysis while the ileal and colonic contents were collected for 16S rRNA gene sequencing. The results showed HFD significantly increased mice body weight, epididymal fat weight, perirenal fat weight, liver weight, and the lipid content in the liver ( P < 0.05). The relative expression of ANGPTL4 and the ANGPTL4-positive cells in the ileum and colon of mice was significantly increased by HFD treatment. Furthermore, 16S rRNA gene sequencing of the ileal and colonic microbiota suggested that HFD treatment changed the composition of the gut microbiota. The ratio of Firmicutes to Bacteroidetes and the abundance of Allobaculum was significantly higher in the HFD group than in the Control group while the abundance of Adlercreutzia, Bifidobacterium, Prevotellaceae UCG-001 , and Ruminococcus was significantly decreased. Interestingly, the abundance of Allobaculum was positively correlated with the expression of ANGPTL4. These findings provide a theoretical foundation for the development of strategies to control the obesity and related diseases by the regulation of ANGPTL4 and gut microbiota.
Hyaluronic acid (HA) is a large non-sulfated glycosaminoglycan that is the main component of the extracellular matrix (ECM). Because of its strong and diversified functions applied in broad fields, HA has been widely studied and reported previously. The molecular properties of HA and its derivatives, including a wide range of molecular weights but distinct effects on cells, moisture retention and anti-aging, and CD44 targeting, promised its role as a popular participant in tissue engineering, wound healing, cancer treatment, ophthalmology, and cosmetics. In recent years, HA and its derivatives have played an increasingly important role in the aforementioned biomedical fields in the formulation of coatings, nanoparticles, and hydrogels. This article highlights recent efforts in converting HA to smart formulation, such as multifunctional coatings, targeted nanoparticles, or injectable hydrogels, which are used in advanced biomedical application.