Implications on growth performance, glucose metabolism, PI3K/AKT pathway, intestinal flora induced by dietary taurine in a high-carbohydrate diet for grass carp (Ctenopharyngodon idella).

An 8-week experiment was performed to investigate the influence on growth performance, plasma biochemistry, glucose metabolism and the insulin pathway of supplementation of dietary taurine to a high-carbohydrate diet for grass carp. In this study, fish were fed diets at one of two carbohydrate levels, 31·49 % (positive control) or 38·61 % (T00). The high-carbohydrate basal diet (T00), without taurine, was supplemented with 0·05 % (T05), 0·10 % (T10), 0·15 % (T15) or 0·20 % (T20) taurine, resulting in six isonitrogenous (30·37 %) and isolipidic (2·37 %) experimental diets. The experimental results showed that optimal taurine level improved significantly weight gain, specific growth rate (SGR), feed utilisation, reduced plasma total cholesterol levels, TAG and promoted insulin-like growth factor level. Glucokinase, pyruvate kinase and phosphoenolpyruvate carboxykinase activities showed a quadratic function model with increasing dietary taurine level, while hexokinase, fatty acid synthetase activities exhibited a positive linear trend. Optimal taurine supplementation in high-carbohydrate diet upregulated insulin receptor (Ir), insulin receptor substrate (Irs1), phosphatidylinositol 3-kinase (pi3k), protein kinase B (akt1), glycogen synthase kinase 3 ß (gs3kß) mRNA level and downregulated insulin-like growth factor (igf-1), insulin-like growth factor 1 receptor (igf-1R) and Fork head transcription factor 1 (foxo1) mRNA level. The above results suggested that optimal taurine level could improve growth performance, hepatic capacity for glycolipid metabolism and insulin sensitivity, thus enhancing the utilisation of carbohydrates in grass carp. Based on SGR, dietary optimal tributyrin taurine supplementation in grass carp was estimated to be 0·08 %.

An integrated study of glutamine alleviates enteritis induced by glycinin in hybrid groupers using transcriptomics, proteomics and microRNA analyses.

Glutamine has been used to improve intestinal development and immunity in fish. We previously found that dietary glutamine enhances growth and alleviates enteritis in juvenile hybrid groupers (Epinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂). This study aimed to further reveal the protective role of glutamine on glycinin-induced enteritis by integrating transcriptome, proteome, and microRNA analyses. Three isonitrogenous and isolipidic trial diets were formulated: a diet containing 10% glycinin (11S group), 10% glycinin diet supplemented with 2% alanine-glutamine (Gln group), and a diet containing neither glycinin nor alanine-glutamine (fishmeal, FM group). Each experimental diet was fed to triplicate hybrid grouper groups for 8 weeks. The analysis of intestinal transcriptomic and proteomics revealed a total of 570 differentially expressed genes (DEGs) and 169 differentially expressed proteins (DEPs) in the 11S and FM comparison group. Similarly, a total of 626 DEGs and 165 DEPs were identified in the Gln and 11S comparison group. Integration of transcriptome and proteome showed that 117 DEGs showed consistent expression patterns at both the transcriptional and translational levels in the Gln and 11S comparison group. These DEGs showed significant enrichment in pathways associated with intestinal epithelial barrier function, such as extracellular matrix (ECM)-receptor interaction, tight junction, and cell adhesion molecules (P < 0.05). Further, the expression levels of genes (myosin-11, cortactin, tenascin, major histocompatibility complex class I and II) related to these pathways above were significantly upregulated at both the transcriptional and translational levels (P < 0.05). The microRNA results showed that the expression levels of miR-212 (target genes colla1 and colla2) and miR-18a-5p (target gene colla1) in fish fed Gln group were significantly lower compared to the 11S group fish (P < 0.05). In conclusion, ECM-receptor interaction, tight junction, and cell adhesion molecules pathways play a key role in glutamine alleviation of hybrid grouper enteritis induced by high-dose glycinin, in which miRNAs and target mRNAs/proteins participated cooperatively. Our findings provide valuable insights into the RNAs and protein profiles, contributing to a deeper understanding of the underlying mechanism for fish enteritis.

[Advantage analysis of flow-through cell method in quality evaluation of Chinese patent medicine: a case study of Danshen Tablets].

To explore the quality consistency evaluation method for multi-component traditional Chinese medicine and establish a dissolution evaluation method suitable for the characteristics of multi-component Chinese patent medicine, this study discussed the characteristics and advantages of the flow-through cell method in the dissolution evaluation of Chinese patent medicine by comparing the impact of the small cup method and the flow-through cell method on the dissolution behavior of water-soluble and lipid-soluble major active components of Danshen Tablets. Dissolution tests were performed using the small cup method as described in the 2020 edition of the Chinese Pharmacopoeia and the newly introduced flow-through cell method(closed-loop method) with water solution containing 0.5% SDS as dissolution medium. Cumulative dissolution curves of the water-soluble component salvianolic acid B and the lipid-soluble component tanshinone Ⅱ_A in Danshen Tablets were plotted, and fitting and similarity analysis of the dissolution models was conducted to identify the characteristics and advantages of the flow-through cell method. For the small cup method, 150 mL of water containing 0.5% SDS was used as the dissolution medium, with a rotation speed of 75 r·min~(-1) and a temperature of(37±0.5) ℃, and 3 mL of samples were taken at 15, 30 min, 1, 2, and 4 h, with fresh dissolution medium added at the same temperature and volume. For the flow-through cell method, a closed-loop system was used. Danshen Tablets were placed in the flow-through cell with approximately 6.7 g of glass beads, and 150 mL of water containing 0.5% SDS was used as the dissolution medium. The flow rate was set at 20 mL·min~(-1), and the temperature and sampling were the same as the small cup method. The results showed that compared with the small cup method, the flow-through cell method had stronger discriminative power and higher sensitivity in distinguishing the dissolution behavior of the two components, and could better reflect the differences in formulation quality, especially for water-insoluble lipid-soluble components. Given that there were no essential differences in the in vitro release kinetics between the two methods, the flow-through cell method could not only replace the traditional small cup method but also better guide the formulation development and identify quality issues of formulations.

From nitrate to NO: potential effects of nitrate-reducing bacteria on systemic health and disease.

Current research has described improving multisystem disease and organ function through dietary nitrate (DN) supplementation. They have provided some evidence that these floras with nitrate (NO3-) reductase are mediators of the underlying mechanism. Symbiotic bacteria with nitrate reductase activity (NRA) are found in the human digestive tract, including the mouth, esophagus and gastrointestinal tract (GT). Nitrate in food can be converted to nitrite under the tongue or in the stomach by these symbiotic bacteria. Then, nitrite is transformed to nitric oxide (NO) by non-enzymatic synthesis. NO is currently recognized as a potent bioactive agent with biological activities, such as vasodilation, regulation of cardiomyocyte function, neurotransmission, suppression of platelet agglutination, and prevention of vascular smooth muscle cell proliferation. NO also can be produced through the conventional L-arginine-NO synthase (L-NOS) pathway, whereas endogenous NO production by L-arginine is inhibited under hypoxia-ischemia or disease conditions. In contrast, exogenous NO3-/NO2-/NO activity is enhanced and becomes a practical supplemental pathway for NO in the body, playing an essential role in various physiological activities. Moreover, many diseases (such as metabolic or geriatric diseases) are primarily associated with disorders of endogenous NO synthesis, and NO generation from the exogenous NO3-/NO2-/NO route can partially alleviate the disease progression. The imbalance of NO in the body may be one of the potential mechanisms of disease development. Therefore, the impact of these floras with nitrate reductase on host systemic health through exogenous NO3-/NO2-/NO pathway production of NO or direct regulation of floras ecological balance is essential (e.g., regulation of body homeostasis, amelioration of diseases, etc.). This review summarizes the bacteria with nitrate reductase in humans, emphasizing the relationship between the metabolic processes of this microflora and host systemic health and disease. The potential effects of nitrate reduction bacteria on human health and disease were also highlighted in disease models from different human systems, including digestive, cardiovascular, endocrine, nervous, respiratory, and urinary systems, providing innovative ideas for future disease diagnosis and treatment based on nitrate reduction bacteria.

Glutamine improves growth and intestinal health in juvenile hybrid groupers fed high-dose glycinin.

Glutamine addition can improve immunity and intestinal development in fish. This study examined the protective roles of glutamine on growth suppression and enteritis induced by glycinin in juvenile hybrid groupers (female Epinephelus fuscoguttatus × male Epinephelus lanceolatus). The experiment set four isonitrogenous and isolipidic trial diets: a diet containing 10% glycinin (11S), 10% of 11S diet supplemented with 1% or 2% alanine-glutamine (1% or 2% Ala-Gln), and a diet containing neither 11S nor Ala-Gln (FM). A feeding trial was conducted in hybrid grouper for 8 weeks. Weight gain and specific growth rates in Groups 1% and 2% Ala-Gln were significantly higher than those of the 11S group but were similar to those of the FM group. The intestinal muscular layer thickness, plica height and width of the 2% Ala-Gln group were significantly higher than those of Group 11S. The enterocyte proliferation efficiency of the 11S group was significantly lower compared to other groups. Compared with the 11S group, Groups 1% and 2% Ala-Gln fish had increased intestinal lysozyme activities, complement 3 and immunoglobulin M as well as cathelicidin contents. The mRNA levels of tnf-α, il-1ß, ifn-α, and hsp70 genes were more downregulated in Groups 1% and 2% Ala-Gln than in Group 11S. Compared with FM group, fish from the 11S group had significantly lower mRNA levels of myd88, ikkß, and nf-κb p65 genes. These three values in the 2% Ala-Gln group were significantly lower than those in Group 11S but not significantly different from those of Group FM. The relative abundance of Vibrio in Group 11S was higher than that in Groups FM and 2% Ala-Gln. Intestinal glutamine, glutaminase, glutamic acid, α-ketoglutarate, malate dehydrogenase and ATP contents were higher in Groups 1% and 2% Ala-Gln than in Group 11S. These results suggest that glutamine is a useful feed additive to enhance growth and intestinal immunity, alleviate inflammation, and modulate gut microbiota in hybrid grouper fed high-dose glycinin.

Quality Marker Discovery and Quality Evaluation of Eucommia ulmoides Pollen Using UPLC-QTOF-MS Combined with a DPPH-HPLC Antioxidant Activity Screening Method.

Pollen, as an important component of Eucommia ulmoides (EUP), is rich in nutrients and is receiving increasing attention. At present, there are no reports on research related to the chemical composition and quality standards of EUP, and there are significant quality differences and counterfeit phenomena in the market. This study used a UPLC-QTOF-MS system to identify 49 chemical components in EUP for the first time. In the second step, 2,2-diphenyl-1-picrylhydrazyl (DPPH)-HPLC antioxidant activity screening technology was used to identify the main active components of EUP, quercetin-3-O-sophoroside (QSH), quercetin-3-O-sambubioside (QSB), and quercetin 3-O-neohesperidoside (QNH), and their purification, preparation, and structure identification were carried out. Third, molecular docking was used to predict the activity of these components. Fourth, the intracellular ROS generation model of RAW264.7 induced by H2O2 was used to verify and evaluate the activity of candidate active ingredients to determine their feasibility as Q-markers. Finally, a quality control method for EUP was constructed using the three selected components as Q-markers. The identification of chemical components and the discovery, prediction, and confirmation of characteristic Q-markers in EUP provide important references for better research on EUP and the effective evaluation and control of its quality. This approach provides a new model for the quality control of novel foods or dietary supplements.

Dietary flaxseed oil and vitamin E improve semen quality via propionic acid metabolism.

Introduction: Flaxseed oil (FO) and vitamin E (VE) both have antioxidant effects on sperm. The present study investigated the effects of dietary supplementation with FO and/or VE on semen quality. Methods: 16 fertile Simmental bulls were selected and randomly divided into 4 groups (n = 4): the control group (control diet), FO group (control diet containing 24 g/kg FO), VE group (control diet containing 150 mg/kg VE) and FOVE group (control diet containing 150 mg/kg VE and 24 g/kg FO), and the trial lasted 10 weeks. Results: The results showed that the addition of FO independently can increase sperm motion parameters, the levels of catalase (CAT), glutathione peroxidase (GSH-Px), testosterone (T) and estradiol (E2), while reduce oxidative stress in seminal plasma (P < 0.05). Supplement of VE independently can increased the motility, motility parameters, CAT and superoxide dismutase (SOD) levels, and reduce oxidative stress in seminal plasma (P < 0.05). There was an interaction effect of FO × VE on motility and reactive oxygen species (ROS), while GSH-Px and ROS were affected by week × VE 2-way interaction, levels of T and E2 were also affected by the dietary FO × week interaction (P < 0.05). The triple interaction effects of FO, VE and week were significant for malondialdehyde (MDA) (P < 0.05). Compared with the control group, sperm from the FOVE group had a significantly higher in vitro fertilization (IVF) rate, and subsequent embryos had increased developmental ability with reduced ROS levels at the eight-cell stage, then increased adenosine triphosphate (ATP) content and gene expression levels of CAT, CDX2, Nanog, and SOD at the blastocyst stage (P < 0.05). Metabolomic and transcriptomic results indicated that dietary supplementation of FO and VE increased the expression of the metabolite aconitic acid, as well as the expression of ABAT and AHDHA genes. Conclusion: With in-silico analysis, it can be concluded that the effects of dietary FO and VE on improving semen quality and embryo development may be related to increased aconitic acid via the ABAT and AHDHA genes involved in the propionic acid metabolism pathway.

The isolation, identification, whole-genome sequencing of Clostridium butyricum LV1 and its effects on growth performance, immune response, and disease-resistance of Litopenaeus vannamei.

In this study, a strain of Clostridium butyricum was isolated from the intestine of Litopenaeus vannamei with the method of anaerobic microbial isolation and culture. Next, the probiotic properties of LV1 were evaluated with susceptibility tests, tolerance tests, and whole genome sequencing in vivo and in vitro, followed by the analysis of the effect of LV1 on the growth performance, immune response, and disease resistance of Litopenaeus vannamei. According to the results, the 16 S rDNA sequence of LV1 was 100% homolofgous to the reference sequence of Clostridium butyricum. Moreover, LV1 was resistant to several antibiotics including amikacin, streptomycin, and gentamicin and highly tolerated artificial gastric and artificial intestinal fluids. The whole genome of LV1 was 4625,068 bp in size and included 4336 coding genes. Among these genes, GO, KEGG, and COG databases exhibited the highest number of genes annotated to metabolic pathway classes and 105 genes annotated as glycoside hydrolases. Meanwhile, 176 virulence genes were predicted. The use of diets supplemented with 1.2 × 109 CFU/kg of LV1 live cells significantly increased the weight gain and specific growth rates of Litopenaeus vannamei and the activity of serum superoxide dismutase, glutathione peroxidase, acid phosphatase, and alkaline phosphatase (P < 0.05). Meanwhile, the use of these diets markedly improved the relative expression of intestinal immunity- and growth-related genes. In conclusion, LV1 has excellent probiotic properties. Specifically, the addition of 1.2 × 109 CFU/kg of LV1 live cells to the diet improved the growth performance, immune response, and disease-resistance of Litopenaeus vannamei.

Analyzing the Role of Septin9 Gene Methylation in the Diagnosis and Treatment of Primary Liver Cancer in the Elderly.

Context: Because the early symptoms of primary hepatocellular carcinoma (PHC) aren't significant, it's difficult to diagnose it by routine inspection clinically, and if the lesion's diameter is small, less than 2.0 cm, false negatives can occur in pathological examinations. Researchers need to actively search for more diagnostic methods. Objective: The study intended to detect and analyze the value of plasma Septin9 gene methylation for the diagnosis and therapeutic monitoring of PHC in older adults. Design: The research team performed a prospective controlled study. Setting: The study took place at the First Hospital of Qiqihar, an Affiliated Qiqihar Hospital at Southern Medical University in Qiqihar, China. Participants: Participants were 32 patients with PHC and 28 with cholangiocarcinoma (CCA) who had been admitted to the hospital between January 2021 and July 2022 and 40 healthy individuals. Groups: The research team divided participants into three groups: (1) patients with PHC, the PHC group; (2) patients with CCA, the CCA group; and (3) healthy individuals, the control group. Outcome Measures: The research team: (1) determined the positive expression rate of Septin9 gene methylation; (2) measured liver function indicators-alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum total bilirubin (TBIL), direct bilirubin (DBIL), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (GGT), albumin (ALB); and (3) measured tumor markers-alpha-fetoprotein (AFP), carbohydrate antigen (CA) 199, CA125, and CA153. The team also: (1) established a binary logistic regression model based on levels of GGT and plasma Septin9 gene methylation to analyze risk factors and diagnosis accuracy, (2) created a receiver operating characteristic (ROC) curve to analyze diagnostic values; and (3) during followup, analyzed the negative conversion rate of Septin9 gene methylation in participants. Results: The positive expression rate of Septin9 methylation in the PHC group was significantly lower than that that of the CCA group and significantly higher than that of the control group (P < .05). The PHC group's ALT, AST, TBIL, DBIL, ALP, and GGT were significantly higher than those of the control group but significantly lower than those of the CCA group (all P < .05). PHC group's ALB was significantly lower than that of the control group (P < .05). The PHC group's AFP, CA199, and CA125 were significantly higher than those of the control group, and the PHC group's CA199 and CA125 were significantly lower than those in the CCA group (all P < .05). The positive expression of Septin9 gene methylation and the high expression of GGT were risk factors for PHC (OR>1, P < .05). The AUC of the Septin9 gene methylation, the GGT level, and the combined detection of both variables (all AUC > 0.70), suggests that the variables have a diagnostic value in the detection of PHC, with the combined detection having the highest value. The negative conversion rate after surgery of Septin9 gene methylation was 87.10%, for 27 out of 31 participants in the PHC and CCA groups (χ2 = 29.405, P < .001). Conclusion: Plasma Septin9 gene methylation is a sensitive molecular marker for the diagnosis and therapeutic monitoring of older adults with PHC, and combined with the serum GGT level, has a high diagnostic efficiency, which may reflect the treatment status of patients.

Salidroside alleviates dexamethasone-induced inhibition of bone formation via transforming growth factor-beta/Smad2/3 signaling pathway.

Glucocorticoid-induced osteoporosis is the third epidemic osteoporosis following postmenopausal and senileosteoporosis. According to one study, salidroside made ovariectomized rats' bones strong. Salidroside's potential for treating glucocorticoid-induced osteoporosis remains unproven. This study aimed to investigate the protective effect and mechanism of salidroside on dexamethasone-induced osteogenic differentiation and bone formation in MC3T3-E1 cells and zebrafish. The study proved that salindroside had no harmful impact on MC3T3E1 cells. Salidroside significantly relieved dexamethasone-induced inhibition of ALP (alkaline phosphatase) activity and mineralization in MC3T3-E1 cells, and promoted osteogenic differentiation of cells. Salidroside increased the expression of osteopontin (OPN), runt-related transcription factor 2 (Runx2), osterix (Osx), transforming growth factor-beta (TGF-ß) proteins and promoted the phosphorylation of Smad2/3 in MC3T3-E1 cells treated with dexamethasone. In addition, the effect of salidroside in relieving dexamethasone-induced inhibition of osteogenic differentiation in MC3T3-E1 cells can be blocked by TGF-ß receptor type I/II inhibitor (LY2109761). At the same time, we found that salidroside significantly alleviated the inhibition of dexamethasone-induced bone formation in zebrafish and promoted the mineralization of zebrafish skulls. LY2109761 reversed the protective impact of salidroside on dexamethasone-mediated bone impairment in zebrafish. These findings suggested that salidroside alleviated dexamethasone-induced inhibition of osteogenic differentiation and bone formation via TGF-ß/Smad2/3 signaling pathway.

Dietary phosphorus supplementation in the diet of Pacific white shrimp (Litopenaeus vannamei) alleviated the adverse impacts caused by high Clostridium autoethanogenum protein.

The study evaluated the effects of dietary phosphorus supplementation on the fishmeal replacement with Clostridium autoethanogenum protein (CAP) in the diet of L. vannamei. Four isonitrogenous and isolipid diets were formulated: the PC diet contains 25% fishmeal, the NC, P1 and P2 diets were replaced 40% fishmeal with CAP and supplemented with 0, 0.8 and 1.6% NaH2PO4 respectively (equivalent to dietary phosphorus level of 0.96%, 1.12% and 1.27%). Sampling and V. parahaemolyticus challenge test were conducted after 50-day-feeding (initial shrimp weight 1.79 ± 0.02 g). The results showed that there were no significant differences in the growth performance of shrimp among the 4 groups. The expressions of dorsal in the gut were significantly lower in shrimp fed the P1 and P2 diets than shrimp fed the NC diet and the expression of peroxinectin in the gut was lower in shrimp fed the NC diet than others. The cumulative mortality of shrimp after V. parahaemolyticus challenge was significantly lower in shrimp fed the P2 diet than those fed the NC diet. After the challenge, genes expressions related to the prophenoloxidase activating system (proPO, lgbp, ppaf) were inhibited in the hepatopancreas of shrimp fed NC diet but activated in shrimp fed the P1 diet compared to those fed the PC diet. The AKP and T-AOC activities were higher in shrimp fed the P2 diet than those fed the other diets. The thickness of muscle layer of shrimp fed the P1 diet was thicker than that in the other groups, and significant stress damage happened in the midgut of the shrimp fed the NC diet. The abundance of Pseudoalteromonas, Haloferula and Ruegeria in shrimp fed the P1 diet was higher than those fed the other diets, while Vibrio in shrimp fed the P2 diet was higher than those fed the other diets. This indicated that a low fishmeal diet with dietary phosphorus level of 1.12% could improve the histology, enhance immune response, and increase the abundance of beneficial bacteria in the gut of shrimp. The low fishmeal diet with dietary phosphorus level of 1.27% could improve disease resistance and antioxidant capacity, but there was a possibility of damage to the gut histology as well as increasing abundance of Vibrio in the gut microbiota of shrimp.

Supplementing artemisinin positively influences growth, antioxidant capacity, immune response, gut health and disease resistance against Vibrio parahaemolyticus in Litopenaeus vannamei fed cottonseed protein concentrate meal diets.

Artemisinin (ART) is a kind of Chinese herbal medicine worth exploring, which obtains various physiological activities. In order to study the prebiotic effect of ART on Litopenaeus vannamei fed cottonseed protein concentrate meal diets, six groups of isonitrogenous and isolipid diets were prepared (including the fish meal control group, FM; cottonseed protein concentrate replacing 30% fishmeal protein and supplementing ART groups: ART0, ART0.3, ART0.6, ART0.9, and ART1.2). The feeding trials was lasted for 56 days. The results showed that the final body weight, weight gain and specific growth rate of the ART0.6 group were the highest, yet the feed coefficient rate of the ART0.6 group was the lowest significantly (P < 0.05). There was no significant difference in survival rate among treatments (P > 0.05). In serum, the content of malondialdehyde in ART0 group was the highest (P < 0.05); the activities of superoxide dismutase, catalase, phenol oxidase and lysozyme increased firstly and then decreased among the ARTs groups (P < 0.05). The activities of intestinal digestive enzymes (including the trypsin, lipase and amylase) showed an upward trend among the ARTs groups (P < 0.05). The histological sections showed that the intestinal muscle thickness, fold height and fold width in the FM group were significantly better than those in the ART0 group; while the mentioned above morphological indexes in the ART0 group were significantly lowest among the ARTs groups (P < 0.05). Sequencing of intestinal microbiota suggested that the microbial richness indexes firstly increased and then decreased (P < 0.05); the bacterial community structure of each treatment group was almost close; the relative abundance of pathogenic bacteria decreased significantly (P < 0.05), such as the Proteobacteria and Cyanobacteria at phylum level, besides the Vibrio and Candidatus Bacilloplasma at genus level. In intestinal tissue, the relative expression levels of TOLL1, TRAF6 and Pehaeidih3 showed up-regulated trends, while the expression of Crustin and LZM firstly up-regulated and then down-regulated (P < 0.05). The challenge experiment suggested that the cumulative mortality of FM group was significantly lower than that of ART0 group; besides the cumulative mortality firstly increased and then decreased between the ARTs groups (P < 0.05). In conclusion, the dietary supplementation of ART can improve the growth, antioxidant capacity, immune response, gut health and disease resistance of the shrimp. To be considered as a dietary immune enhancer, the recommended supplementation level of ART in shrimp's cottonseed protein concentrate meal diets is 0.43%.

Baogong decoction treats endometritis in mice by regulating uterine microbiota structure and metabolites.

Endometritis is persistent inflammation caused by bacteria, which can lead to infertility. Although traditional Chinese medicine (TCM) has been used to treat endometritis, the underlying mechanism is still unclear. Here, Baogong Decoction (BGD), a TCM compound, was used to treat mouse endometritis induced by Escherichia coli (E. coli), and then 16S rRNA sequencing and non-targeted metabolomics were used to investigate the change of uterine microbiota and metabolomes in serum and uterine after BGD treatment. Finally, the therapeutic effect of potential metabolites for treating mouse endometritis screened by combined omics analyses was verified using pathological model. The results showed that BGD treatment could effectively treat endometritis associated with the increasing relative abundance of Firmicutes, Bacteroides, Lactobacillus and Lactococcus, and the decreasing relative abundance of Cupriavidus and Proteobacteria. 133 and 130 metabolites were found to be potential biomarkers in serum and uterine tissue respectively. In serum and tissues, dehydroepiandrosterone (DHEA) and catechol were significantly increased in the BGD treatment versus the inflammation group. Results of combined omics analyses demonstrated that DHEA was positively correlated with changes in microbiota. Results of pathological model demonstrated that DHEA could cure endometritis effectively associated with the decreasing infiltration of inflammatory cells and expression of inflammatory factors in the uterus. In summary, our results demonstrated that BGD could cure endometritis in mice by modulating the structure of the uterine microbiota and its metabolites, in which DHEA may be one of the main components of the therapeutic effect of BGD.

Compound glycyrrhiza oral solution alleviates oxidative stress and inflammation by regulating SRC/MAPK pathway in chronic obstructive pulmonary disease.

OBJECTIVES: Patients with chronic obstructive pulmonary disease (COPD) suffer from persistent cough and breathlessness, which can be ameliorated by the Chinese herbal medicine glycyrrhiza. Furthermore, the SRC/MAPK pathway is activated in the process of oxidative stress and inflammation, which afflict COPD progression. Thus, this research aimed at dissecting the mechanism of compound glycyrrhiza oral solution (CGOS) relieving oxidative stress and inflammation in COPD via the SRC/MAPK pathway. METHODS: After a COPD rat model was established using lipopolysaccharide and cigarette smoke, rats underwent intragastric administration with CGOS and intratracheal injection with LV-NC and LV-SRC lentivirus into lungs. Then, pulmonary function-related indexes were evaluated, followed by analyses of arterial blood and inflammatory cell number in prepared bronchoalveolar lavage fluids. Meanwhile, the contents of oxidative stress-related indicators (malondialdehyde, 3NT, 8-Isoprostane, glutathione, NO, and SOD) in pulmonary tissues were measured, along with RT-qPCR and ELISA detection of the expression of inflammatory factors (TNF-α, IL-1ß, IL-4, and IL-10). Moreover, western blot assay was utilized to assess p-SRC/SRC and p-p38/p38 ratios in pulmonary tissues. RESULTS: CGOS treatment enhanced PaO2 and reduced PaCO2 in COPD rats, accompanied by declines in the number of total cells, neutrophils, and macrophages. CGOS improved pulmonary function, decreased malondialdehyde, 3NT, 8-Isoprostane, TNF-α, and IL-1ß levels, and increased GSH, NO, IL-4, and IL-10 levels and SOD activity. Mechanistically, CGOS suppressed the SRC/MAPK pathway, and SRC overexpression reversed the alleviating function of CGOS in COPD rats. CONCLUSIONS: In conclusion, CGOS might alleviate oxidative stress and inflammation in COPD rats by inhibiting the SRC/MAPK pathway.

Effects of the Clostridium butyricum on growth performance, antioxidant capacity, immunity and disease resistance of Litopenaeus Vannamei fed with cottonseed protein concentrate (CPC) replacement of fishmeal in diet.

Clostridium butyricum (CB) is a gram-positive bacterium that secretes short-chain fatty acids such as butyric acid and so on. An 8-week feeding trial was conducted to investigate the effects of CB on the growth performance, antioxidant capacity, immunity and resistance to Vibrio parahaemolyticus in Litopenaeus Vannamei fed with cottonseed protein concentrate (CPC) replacement of fishmeal. Six iso-nitrogenous (40%) and iso-lipidic (6%) diets were formulated including a positive control group (PC, 25% fishmeal), a negative control group (NC, CPC replaced 30% of fishmeal protein), and 0.03% (C1, 3 × 108 CFU/kg), 0.12% (C2, 1.2 × 109 CFU/kg), 0.48% (C3, 4.8 × 109 CFU/kg) and 1.92% (C4, 1.92 × 1010 CFU/kg) CB were supplemented on the negative control group (NC). After the feeding trial, the remaining shrimp in each treatment group were subjected to a challenge experiment with Vibrio parahaemolyticus. The results indicated that weight gain rate (WGR), specific growth rate (SGR) in C4 group were significantly lower than those in PC and C2 groups (P < 0.05); the feed conversion ratio (FCR) was significantly higher than that of PC and C2 groups (P < 0.05). There was no significant difference in survival rate (SR) among all groups (P > 0.05). Compared to the PC and NC groups, the total superoxide capacity, superoxide dismutase and lysozyme were significantly higher in the C4 group (P < 0.05); the glutathione peroxidase, acid phosphatase and alkaline phosphatase were significantly higher in the C3 group (P < 0.05); and the malondialdehyde was significantly lower in the C4 group (P < 0.05). The relative mRNA expressions of Toll receptor (TLR), innate immune deficiency gene (IMD), penaiedin3a (Pen3) were significantly down-regulated in the NC group than those in the PC group (P < 0.05). In addition, the relative mRNA expressions of TLR, IMD and Pen3 were significantly up-regulated in all groups supplemented with CB than those in the NC group (P < 0.05). Moreover, the cumulative mortality rate in the NC group was not significantly different from the PC group (P > 0.05) and was significantly higher than those in the C3 and C4 groups (P < 0.05). In conclusion, the CB supplementation on the basis of CPC replacement of 30% fishmeal protein enhanced significantly the antioxidant capacity, immunity and disease resistance of shrimp and improved its growth performance. Therefore, considering the factors of the growth, immunity and disease resistance, the CB supplementation of 0.12%-0.48% (1.2 × 109 CFU/kg-4.8 × 109 CFU/kg) was recommended in the diet of L. vannamei based on the results of this experiment.

Interactive Effects on Motor Performance of Mindfulness, Performance Under Pressure, Self-Talk, and Motor Task Characteristics.

High mindfulness individuals have been found to perform better on motor tasks under various conditions, but it is unknown whether mindfulness and performance relate when performing under pressure or using different types of self-talk with different motor tasks. In this study, 46 male participants (Mage = 21.4, SD = 1.72 years) with high mindfulness (n = 23) and low mindfulness (n = 23) performed dart-throwing and two-hand coordination tasks under pressure and non-pressure conditions and when using instructional and unrelated self-talk. First, on the two-hand coordination task, a three-way mixed ANOVA found: (a) a significant 3-way interaction in which a significantly poorer performance occurred under pressure (vs. without pressure), with low (vs. high) mindfulness and when using unrelated (vs. instructional) self-talk and (b) a significant interaction in which, both under pressure and not, both high and low mindfulness participants performed comparably when using instructional (vs. unrelated) self-talk. Second, on the dart-throwing task, mindfulness interacted with self-talk such that both high and low mindfulness participants performed better when using instructional self-talk, and pressure interacted with self-talk such that participants using instructional (vs. unrelated) self-talk performed better in both pressure and non-pressure conditions. We concluded that instructional self-talk was a useful cognitive strategy, perhaps particularly in pressure conditions and regardless of the degree of mindfulness, and its effectiveness extended to two different motor tasks. We discussed the theoretical implications of these findings, in terms of attention theory, self-talk, and motor control; and we highlighted our study's limitations and practical applications and gave recommendations for future research.

Combined Effect of Flaxseed Oil and Vitamin E Supplementation During Bull Semen Cryopreservation on Sperm Characteristics.

The present study was performed to investigate the effects of supplementing flaxseed oil (FO) or vitamin E (VE) or their combination to an extender for Simmental bull semen cryopreservation. In experiment 1, different concentrations of FO (0, 10, 100, and 1000 ng/mL) and VE (0.05, 0.1, and 0.2 mg/mL) were added to the extenders. In experiment 2, FO, VE, and FO + VE were added and a control group was included. Sperm viability, motility, motion parameters, acrosome integrity and membrane integrity, endogenous antioxidant indices, reactive oxygen species, and malondialdehyde levels were evaluated after semen thawing. A higher percentage of viability, motion parameters, endogenous antioxidant indices, and membrane integrity was observed after supplementation with 10 ng/mL FO or 0.1 mg/mL VE compared with the control group (p < 0.05). Also, combined supplementation of 10 ng/mL FO +0.1 mg/mL VE further improved the quality of frozen-thawed sperm by regulating viability, motion parameters, membrane integrity, and endogenous antioxidant indices compared with the FO or VE alone (p < 0.05). These results indicated that FO (10 ng/mL) + VE (0.1 mg/mL) could further improve the protective effects on bull sperm post-thaw.

Exploring the efficacy of tumor electric field therapy against glioblastoma: An in vivo and in vitro study.

AIMS: Tumor electric fields therapy (TTFields) is emerging as a novel anti-cancer physiotherapy. Despite recent breakthroughs of TTFields in glioma treatment, the average survival time for glioblastoma patients with TTFields is <2 years, even when used in conjugation with traditional anti-cancer therapies. To optimize TTFields-afforded efficacy against glioblastoma, we investigated the cancer cell-killing effects of various TTFields paradigms using in vitro and in vivo models of glioblastoma. METHODS: For in vitro studies, the U251 glioma cell line or primary cell cultures prepared from 20 glioblastoma patients were treated with the tumor electric field treatment (TEFT) system. Cell number, volume, and proliferation were measured after TEFT at different frequencies (100, 150, 180, 200, or 220 kHz), durations (24, 48, or 72 h), field strengths (1.0, 1.5, or 2.2V/cm), and output modes (fixed or random sequence output). A transwell system was used to evaluate the influence of TEFT on the invasiveness of primary glioblastoma cells. For in vivo studies, the therapeutic effect and safety profiles of random sequence electric field therapy in glioblastoma-transplanted rats were assessed by calculating tumor size and survival time and evaluating peripheral immunobiological and blood parameters, respectively. RESULTS: In the in vitro settings, TEFT was robustly effective in suppressing cell proliferation of both the U251 glioma cell line and primary glioblastoma cell cultures. The anti-proliferation effects of TEFT were frequency- and "dose" (field strength and duration)-dependent, and contingent on the field sequence output mode, with the random sequence mode (TEFT-R) being more effective than the fixed sequence mode (TEFT-F). Genetic tests were performed in 11 of 20 primary glioblastoma cultures, and 6 different genetic traits were identified them. However, TEFT exhibited comparable anti-proliferation effects in all primary cultures regardless of their genetic traits. TEFT also inhibited the invasiveness of primary glioblastoma cells in transwell experiments. In the in vivo rat model of glioblastoma brain transplantation, treatment with TEFT-F or TEFT-R at frequency of 200 kHz and field strength of 2.2V/cm for 14 days significantly reduced tumor volume by 42.63% (TEFT-F vs. control, p = 0.0002) and 63.60% (TEFT-R vs. control, p < 0.0001), and prolonged animal survival time by 30.15% (TEFT-F vs. control, p = 0.0415) and 69.85% (TEFT-R vs. control, p = 0.0064), respectively. The tumor-bearing rats appeared to be well tolerable to TEFT therapies, showing only moderate increases in blood levels of creatine and red blood cells. Adverse skin reactions were common for TEFT-treated rats; however, skin reactions were curable by local treatment. CONCLUSION: Tumor electric field treatment at optimal frequency, strength, and output mode markedly inhibits the cell viability, proliferation, and invasiveness of primary glioblastoma cells in vitro independent of different genetic traits of the cells. Moreover, a random sequence electric field output confers considerable anti-cancer effects against glioblastoma in vivo. Thus, TTFields are a promising physiotherapy for glioblastoma and warrants further investigation.

Dietary supplementation of ß-conglycinin, with or without sodium butyrate on the growth, immune response and intestinal health of hybrid grouper.

We investigated the effects of low and high doses of ß-conglycinin and the ameliorative effects of sodium butyrate (based on high-dose ß-conglycinin) on the growth performance, serum immunity, distal intestinal histopathology, and gene, protein expression related to intestinal health in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂). The results revealed that the instantaneous growth rate (IGR) of grouper significantly increased, decreased, and increased in the low-dose ß-conglycinin (bL), high-level ß-conglycinin (bH) and high-level ß-conglycinin plus sodium butyrate (bH-NaB), respectively. The feed coefficient ratio (FCR) was significantly increased in the bH and bH-NaB, serum levels of IFN-γ, IL-1ß, and TNF-α were upregulated in the bH. The intestinal diameter/fold height ratio was significantly increased in the bH. Furthermore, there were increases in nitric oxide (NO), total nitric oxide synthase (total NOS), and peroxynitrite anion (ONOO-) in the bH, and decreases in total NOS and ONOO- in the bH-NaB. In the distal intestine, IL-1ß and TGF-ß1 mRNA levels were downregulated and upregulated, respective in the bL. The mRNA levels of TNF-α and IL-6 were upregulated in the bH, and downregulated in the bH-NaB, respectively. Occludin, claudin3 and ZO-3 mRNA levels were upregulated in the bL, downregulated in the bH and then upregulated in the bH-NaB. No significant differences were observed in the mRNA levels of IFN-γ and jam4. And the p-PI3K p85Tyr458/total PI3K p85 value was significantly increased in the bH and then decreased in the bH-NaB, and the total Akt value was significantly increased in the bH. These indicate ß-conglycinin has a regulatory effect on serum immunity and affect distal intestinal development by modulating distal intestinal injury-related parameters. Within the distal intestinal tract, low- and high-dose ß-conglycinin differentially affect immune responses and tight junctions in the distal intestine, which eventually manifests as a reduction in growth performance. Supplementing feed with sodium butyrate might represent an effective approach for enhancing serum immunity, and protects the intestines from damage caused by high-dose ß-conglycinin.