Dietary Zn Deficiency Inhibits Cell Proliferation via the GPR39-Mediated Suppression of the PI3K/AKT/mTOR Signaling Pathway in the Jejunum of Broilers.

A prior investigation revealed that a lack of Zinc (Zn) could hinder intestinal cell proliferation in broiler chickens; however, the mechanisms responsible for this effect remain unclear. We aimed to investigate the possible mechanisms of dietary Zn deficiency in inhibiting the jejunal cell proliferation of broilers. For this study, a total of 112 chickens (21 days old) were randomly divided into two treatments (seven replicate cages per treatment, eight chickens per replicate cage): the control group (CON) and the Zn deficiency group. The duration of feeding was 21 d. Chickens in the control group were provided with a basal diet containing an extra addition of 40 mg Zn/kg in the form of Zn sulfate, whereas chickens in the Zn deficiency group were given the basal diet with no Zn supplementation. The results indicated that, in comparison to the CON, Zn deficiency increased (p < 0.05) the duodenal and jejunal crypt depth (CD) of broilers on d 28 and jejunal and ileal CD on d 35, and decreased (p < 0.05) the duodenal, jejunal, and ileal villus height/crypt depth (VH/CD) on d 28 and the jejunal VH, jejunal and ileal villus surface area, and VH/CD on d 35. Furthermore, Zn deficiency decreased (p < 0.0001) the number of proliferating cell nuclear antigen-positive cells and downregulated (p < 0.01) the mRNA or protein expression levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K, phosphorylated serine-threonine kinase (AKT), phosphorylated mechanistic target of rapamycin (mTOR), G protein-coupled receptor 39 (GPR39), and extracellular-regulated protein kinase, but upregulated (p < 0.05) the mRNA or protein expression levels of P38 mitogen-activated protein kinase, c-jun N-terminal kinase (JNK) 1 and JNK2, and phosphorylated protein kinase C in the jejunum of the broilers on d 42. It was concluded that dietary Zn deficiency inhibited cell proliferation possibly via the GPR39-mediated suppression of the PI3K/AKT/mTOR signaling pathway in the jejunum of broilers.

Dietary copper requirement of broilers fed a corn-soybean meal diet during 22-42 d of age.

This research was to assess the dietary copper (Cu) requirement of broiler chickens fed a practical corn-soybean meal diet during 22-42 d of age. A total of 288 numbered Arbor Acres male broilers at 22 d of age were randomly allotted 6 treatments with 8 replicate cages (6 broilers per cage) per treatment. Broilers were fed a Cu-unsupplemented corn-soybean meal basal diet (control, containing 7.36 mg Cu/kg) or the basal diet added with 3, 6, 9, 12, or 15 mg Cu/kg from CuSO4·5H2O for 21 d. Quadratic, asymptotic and broken-line models were fitted and the best fitted models were selected to determine dietary Cu requirements. The results revealed that the contents of Cu in serum and liver, mRNA expression levels of Cu- and zinc-containing superoxide dismutase (CuZnSOD) in liver and monoamine oxidase b (MAO B) in heart, as well as protein expression level of CuZnSOD in liver were affected (P < 0.05) by supplemental Cu levels, and the above indices varied linearly and quadratically (P < 0.05) with increasing Cu levels. Dietary Cu requirements assessed according to the best fitted broken-line models (P < 0.05) of the above indexes were 10.45-13.81 mg/kg. It was concluded that mRNA expression levels of CuZnSOD in liver and MAO B in heart, as well as liver CuZnSOD protein expression level were new specific sensitive biomarkers for estimating dietary Cu requirements, and the dietary Cu requirement was recommended to be 14 mg/kg to support Cu metabolic needs related to key Cu-containing enzymes in broilers fed the corn-soybean meal diet during 22-42 d of age, which was higher than the dietary Cu requirement (8 mg/kg) for broilers at the corresponding stage suggested by the Chinese Feeding Standard of Chicken.

The chemical characteristics of different sodium iron ethylenediaminetetraacetate sources and their relative bioavailabilities for broilers fed with a conventional corn-soybean meal diet.

BACKGROUND: Our previous studies demonstrated that divalent organic iron (Fe) proteinate sources with higher complexation or chelation strengths as expressed by the greater quotient of formation (Qf) values displayed higher Fe bioavailabilities for broilers. Sodium iron ethylenediaminetetraacetate (NaFeEDTA) is a trivalent organic Fe source with the strongest chelating ligand EDTA. However, the bioavailability of Fe when administered as NaFeEDTA in broilers and other agricultural animals remains untested. Herein, the chemical characteristics of 12 NaFeEDTA products were determined. Of these, one feed grade NaFeEDTA (Qf = 2.07 × 108), one food grade NaFeEDTA (Qf = 3.31 × 108), and one Fe proteinate with an extremely strong chelation strength (Fe-Prot ES, Qf value = 8,590) were selected. Their bioavailabilities relative to Fe sulfate (FeSO4·7H2O) for broilers fed with a conventional corn-soybean meal diet were evaluated during d 1 to 21 by investigating the effects of the above Fe sources and added Fe levels on the growth performance, hematological indices, Fe contents, activities and gene expressions of Fe-containing enzymes in various tissues of broilers. RESULTS: NaFeEDTA sources varied greatly in their chemical characteristics. Plasma Fe concentration (PI), transferrin saturation (TS), liver Fe content, succinate dehydrogenase (SDH) activities in liver, heart, and kidney, catalase (CAT) activity in liver, and SDH mRNA expressions in liver and kidney increased linearly (P < 0.05) with increasing levels of Fe supplementation. However, differences among Fe sources were detected (P < 0.05) only for PI, liver Fe content, CAT activity in liver, SDH activities in heart and kidney, and SDH mRNA expressions in liver and kidney. Based on slope ratios from multiple linear regressions of the above indices on daily dietary analyzed Fe intake, the average bioavailabilities of Fe-Prot ES, feed grade NaFeEDTA, and food grade NaFeEDTA relative to the inorganic FeSO4·7H2O (100%) for broilers were 139%, 155%, and 166%, respectively. CONCLUSIONS: The bioavailabilities of organic Fe sources relative to FeSO4·7H2O were closely related to their Qf values, and NaFeEDTA sources with higher Qf values showed higher Fe bioavailabilities for broilers fed with a conventional corn-soybean meal diet.

Phenylboronic acid-functionalized magnetic metal-organic framework nanoparticles for magnetic solid phase extraction of five benzoylurea insecticides.

This research involves the construction of a phenylboronic acid-functionalized magnetic UiO-66 metal-organic framework (MOF) nanoparticle (CPBA@UiO-66@Fe3O4). Its design is primarily for the magnetic solid phase extraction (MSPE) of benzoylurea insecticides. An organic ligand, 2-amino terephthalic acid (2-ATPA), facilitated the introduction of amino groups while keeping the original crystal structure of UiO-66 intact. The constructed UiO-66 MOF showcases a porous structure and extensive surface area, thereby providing an optimal platform for further functionalization. The employment of 4-carboxylphenylboronic acid as a modifier notably amplified the extraction efficiency for benzoylureas. This improvement was due to the formation of B-N coordination and other secondary interactions. By integrating this with high-performance liquid chromatography (HPLC), we established a quantitative analytical method for benzoylurea insecticides. This method achieved a wide linear range (2.5-500 µg L-1 or 5-500 µg L-1), satisfactory recoveries (83.3-95.1%), and acceptable limits of detection (LODs: 0.3-1.0 µg L-1). The developed method proved successful when applied to six tea infusion samples, representing China's six major tea categories. Semi-fermented and light-fermented tea samples demonstrated relatively higher spiking recoveries.

Dietary protocatechuic acid ameliorates ileal mucosal barrier injury and inflammatory response and improves intestinal microbiota composition in Yellow chickens challenged with Salmonella typhimurium.

Salmonella typhimurium (ST) is a common foodborne pathogen that severely affects the health of humans and livestock. Protocatechuic acid (PCA) has been shown to possess anti-inflammatory and anti-bacterial functions. Chickens were used to investigate the effect of PCA on the gut health infected with ST. A total of one hundred eighty, 1-d-old birds were randomly allocated into 3 treatments, each with 6 replicates per treatment and 10 chicks per replicate. Broiler chicks in the control and ST treatment were fed a basal diet, and birds in the PCA+ST treatment received the basal diet with 600 mg/kg PCA. On d 14 and 16 of the trial, broilers in ST and PCA+ST treatments received an oral dose of ST, while broilers in CON received an equal amount of PBS. The data were analyzed by the one-way ANOVA. Dietary PCA increased (P < 0.05) final body weight, average daily gain, and feed to gain ratio in ST-challenged Yellow broilers. Protocatechuic acid significantly alleviated ST-induced intestinal mucosal injury reflected in the decreased (P < 0.05) plasma activity of diamine oxidase and ileal apoptosis, with increased (P < 0.05) ileal villus height and villus height/crypt depth. Protocatechuic acid treatment significantly decreased (P < 0.05) ST-induced proinflammatory cytokine (Interleukin-1ß, Interleukin-6, Tumor necrosis factor-α, and Interferon-ß) content in ileum. Meanwhile, PCA treatment significantly increased (P < 0.05) the transcript abundances of claudin 1 (CLDN1), zonula occludens-1 (ZO-1), and mucin 2 (MUC2) in ileum, all related to the intestinal barrier in ST-challenged Yellow broilers. Additionally, PCA also increased (P < 0.05) the diversity and richness of the cecal microflora as reflected by reduced (P < 0.05) abundance of Bacteroidota, Proteobacteria and Escherichia-Shigella, and increased (P < 0.05) abundance of Firmicutes and Lactobacillus in ST-challenged Yellow broilers. These findings indicate that PCA relieves ST-induced loss weight, intestinal barrier injury, inflammatory response, and improves intestinal microbiota composition in Yellow broilers.

The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers.

Our previous study demonstrated that the zinc (Zn) proteinate with moderate chelation strength (Zn-Prot M) enhanced the Zn absorption in the small intestine partially via increasing the expression of some Zn and amino acid transporters in the duodenum of broilers. However, it remains unknown whether the Zn-Prot M could also regulate the expression of related transporters in the jejunum and ileum of broilers in the above enhancement of Zn absorption. The present study was conducted to investigate the effect of the Zn-Prot M on the expression of related transporters in the jejunum and ileum of broilers compared to the Zn sulfate (ZnS). Zinc-deficient broilers (13-d-old) were fed with the Zn-unsupplemented basal diets (control) or the basal diets supplemented with 60 mg Zn/kg as ZnS or Zn-Prot M for 26 d. The results showed that in the jejunum, compared to the control, supplementation of the organic or inorganic Zn increased (P < 0.05) mRNA and protein expression of b0,+-type amino acid transporter (rBAT), Zn transporter 10 (ZnT10), and peptide-transporter 1 (PepT1) mRNA expression and Zn transporter 7 (ZnT7) protein expression on d 28, while y+L-type amino transporter 2 (y+LAT2) mRNA and protein expression, and protein expression of ZnT7 and ZnT10 on 28 d and zrt-irt-like protein 3 (ZIP3) and zrt-irt-like protein 5 (ZIP5) on d 39 were higher (P < 0.05) for Zn-Prot M than for ZnS. In the ileum, Zn addition regardless of Zn source up-regulated (P < 0.05) mRNA expression of Zn transporter 9 (ZnT9) and ZIP3, ZIP5, and y+LAT2 protein expression on d 28, and PepT1 mRNA and protein expression, ZIP3 and y+LAT2 mRNA expression and ZnT10 protein expression on d 39. Furthermore, Zn transporter 4 (ZnT4) and ZnT9 mRNA expression and Zn transporter 1 (ZnT1) protein expression on d 28, and y+LAT2 mRNA expression and ZnT10 and PepT1 protein expression on d 39 were higher (P < 0.05) for Zn-Prot M than for ZnS. It was concluded that the Zn-Prot M enhanced the expression of the ZnT1, ZnT4, ZnT9, ZnT10, ZIP3, ZIP5, y+LAT2, and PepT1 in the jejunum or ileum of broilers compared to the ZnS.

Dual-mode detection of fluorine-containing pesticides (bifenthrin, flufenoxuron, diflubenzuron) via ratiometric fluorescence and the Tyndall Effect of fluorescent organic nanoparticles.

The Tyndall Effect assay (TEA) has been applied into colorimetric metal ion detection since 2019. However, the TEA-based sensor for pesticide detection has never been reported till now. Herein, a facile fluorescent organic nanoparticle (FON)-based sensor is firstly developed for fluorine-containing pesticide detection through ratiometric fluorescence assay (FLA) and TEA. For FLA, the intensity of the second-order Tyndall scattering peak (STS590nm) and the fluorescence peak of the FON-based sensor would increase and remain unchanged respectively when adding bifenthrin, flufenoxuron, and diflubenzuron. The detection limits were respectively 9.34, 6.91, and 3.60 µg/kg. For TEA, the increased STS590nm intensity displayed a bright and visible light beam. An economical, simple, and portable device was then constructed to visually monitor the analytes. The sensor was successfully used to detect the analytes in teas through FLA and TEA with the recoveries and RSD ranging from 86.27-100.00 %, and 0.00-5.68 %, respectively.

Evaluation of dietary metabolizable energy concentrations on meat quality and lipid metabolism-related gene expression in yellow-feathered chickens.

This study evaluated the effects of different dietary metabolizable energy (ME) concentrations on the meat quality, carcass traits, volatile flavour and lipid metabolism-related gene expression levels in yellow-feathered chickens. In total, 600 Huxu female chickens aged 90 days were randomly assigned to six dietary treatments, each with 10 replicates of 10 birds. During the finisher phase, the birds were fed diets containing 2880 (low), 2940, 3000, 3060, 3120 and 3180 (high) kcal ME/kg. The results showed that the average daily gain of chickens increased as the dietary ME concentration increased, while the feed to gain improved (p < 0.05), and the intramuscular fat content of breast muscle increased (p < 0.05). The energy concentration had no effect on the breast muscle pH (45 min and 24 h), colour parameter (L*) or percentage of drip loss (p > 0.05), but the shear force values decreased significantly (p < 0.05). The diameter and area of the breast muscle fiber decreased and the muscle fibre density increased as the dietary ME concentration increased (p < 0.05). The highest ME concentration (3180 kcal) increased the percentages of aldehydes (hexanal, heptanal, 2,4-nonadienal, octanal, nonanal and 2-decenal), alcohols (2-nonen-1-ol, trans-2-undecen-1-ol, 7-hexadecenal, 2-hexyl-1-decanoal and n-nonadecanol-1,3,7,11-trimethyl-1-dodecanol), alkanes (2,6-dimethyl-heptadecane) and carboxylic acids (9-hexadecenoic acid), but reduced the percentages of octadecanal, octadecane, heneicosane and tetradecanal (p < 0.05). In addition, the mRNA gene expression levels of fatty acid-binding protein 3 and apolipoprotein B were significantly upregulated in the liver, whereas that of cholesteryl ester transfer protein was significantly downregulated. In conclusion, increasing the ME diet to 3180 kcal/kg significantly improved the quality and flavour of the meat from yellow-feathered broilers. our finding may help poultry producers to improve the taste of meat by regulating genes related to lipid metabolism, thereby achieving the flavour and taste characteristics preferred by consumers.

Transforaminal Versus Interlaminar Approach of Full-Endoscopic Lumbar Discectomy Under Local Anesthesia for L5/S1 Disc Herniation: A Randomized Controlled Trial.

BACKGROUND: Local anesthesia is feasible for both transforaminal and interlaminar approaches in percutaneous endoscopic lumbar discectomy (PELD). However, the optimal approach for PELD has not yet been established at the L5/S1 segment under local anesthesia with 1% lidocaine. OBJECTIVES: In this study, we compared the transforaminal approach with the interlaminar approach of PELD under local anesthesia for L5/S1 disc herniation (DH). STUDY DESIGN: This was a prospective randomized clinical trial. METHODS: From January 2019 to March 2020, 91 consecutive patients with L5/S1 DH who planned to undergo PELD in our unit were randomized to the transforaminal endoscopic lumbar discectomy (TELD, n = 46) or interlaminar endoscopic lumbar discectomy (IELD, n = 45). Both procedures were performed under local anesthesia with 1% lidocaine. The clinical outcomes were assessed as the Visual Analog Scale (VAS) score, Oswestry Disability Index (ODI) score, and modified MacNab criteria. Patient satisfaction surveys and surgical complications were also recorded and analyzed. RESULTS: Compared to the IELD group, the TELD group had a shorter operative time and postoperative bed rest time (P < 0.001) but a longer radiation time (P < 0.001) and lower VAS scores for intraoperative back pain (P < 0.001) and leg pain (P < 0.001). At the postoperative follow-up, there were no significant differences between the 2 groups in the VAS scores, ODI scores, or modified MacNab criteria. The surveys showed a significantly higher satisfaction rate in the TELD group than in the IELD group (P = 0.014). Six patients in the IELD group (13.3%) needed extra intravenous injections of sufentanil because of intense pain during the procedure. In the IELD group, there were 2 cases of neuropathic pain after surgery. LIMITATIONS: Due to the study was included in a single spine center with a relatively small population and its relatively short-term follow-up, the study is not generalizable. CONCLUSIONS: Both TELD and IELD can provide good clinical outcomes for L5/S1 DH under local anesthesia with 1% lidocaine. TELD was superior to IELD in terms of surgical-related experience and complications.

Network pharmacology-based strategy for predicting therapy targets of Sanqi and Huangjing in diabetes mellitus.

BACKGROUND: A comprehensive literature search shows that Sanqi and Huangjing (SQHJ) can improve diabetes treatment in vivo and in vitro, respectively. However, the combined effects of SQHJ on diabetes mellitus (DM) are still unclear. AIM: To explore the potential mechanism of Panax notoginseng (Sanqi in Chinese) and Polygonati Rhizoma (Huangjing in Chinese) for the treatment of DM using network pharmacology. METHODS: The active components of SQHJ and targets were predicted and screened by network pharmacology through oral bioavailability and drug-likeness filtration using the Traditional Chinese Medicine Systems Pharmacology Analysis Platform database. The potential targets for the treatment of DM were identified according to the DisGeNET database. A comparative analysis was performed to investigate the overlapping genes between active component targets and DM treatment-related targets. We constructed networks of the active component-target and target pathways of SQHJ using Cytoscape software and then analyzed the gene functions. Using the STRING database to perform an interaction analysis among overlapping genes and a topological analysis, the interactions between potential targets were identified. Gene Ontology (GO) function analyses and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted in DAVID. RESULTS: We screened 18 active components from 157 SQHJ components, 187 potential targets for active components and 115 overlapping genes for active components and DM. The network pharmacology analysis revealed that quercetin, beta-sitosterol, baicalein, etc. were the major active components. The mechanism underlying the SQHJ intervention effects in DM may involve nine core targets (TP53, AKT1, CASP3, TNF, interleukin-6, PTGS2, MMP9, JUN, and MAPK1). The screening and enrichment analysis revealed that the treatment of DM using SQHJ primarily involved 16 GO enriched terms and 13 related pathways. CONCLUSION: SQHJ treatment for DM targets TP53, AKT1, CASP3, and TNF and participates in pathways in leishmaniasis and cancer.

Uncovering the effects and mechanism of Danggui Shaoyao San intervention on primary dysmenorrhea by serum metabolomics approach.

Danggui Shaoyao San (DSS) is a well-known prescription for relieving primary dysmenorrhea (PD) of women in China. However, its pharmacological mechanism has not been thoroughly uncovered. Here, an integrative UPLC-Q-TOF-MS-based serum metabolomics approach coupled with multivariate data analysis has been proposed to investigate the effects and mechanism of DSS on estradiol benzoate and oxytocin-induced PD rats. 31 potential biomarkers of PD were screened and identified, mainly involving phenylalanine, tyrosine and tryptophan biosynthesis, glycerophospholipid metabolism, primary bile acid biosynthesis, and the occurrence of PD could destroy biological homeostasis in vivo by monitoring these pathways. After DSS treatment, 18 identified different metabolites were restored to the nomal state in varying degrees and could be potential biomarkers contributing to the treatment of DSS. These findings implyed that DSS exhibited a therapeutic effect on PD rats through regulating multiple abnormal pathways. Of note, this study discovered some potential biomarkers related to PD for the first time, such as L-tyrosine, glycocholic acid, citric acid, palmitoylcarnitine, cholesterol. It preliminarily proved the pathophysiology of PD and action mechanisms of DSS on PD, and provided a novel insight into the effectiveness of DSS on PD.

Organic zinc with moderate chelation strength enhances zinc absorption in the small intestine and expression of related transporters in the duodenum of broilers.

Our previous study demonstrated that the absorption of zinc (Zn) from the organic Zn proteinate with moderate chelation strength was significantly higher than that of Zn from the inorganic Zn sulfate in the in situ ligated duodenal segment of broilers, but the underlying mechanisms are unknown. The present study aimed to determine the effect of organic Zn with moderate chelation strength and inorganic Zn on the Zn absorption in the small intestine and the expression of related transporters in the duodenum of broilers. The Zn-deficient broilers (13 days old) were fed with the Zn-unsupplemented basal diets (control) containing 25.72 and 25.64 mg Zn/kg by analysis or the basal diets supplemented with 60 mg Zn/kg as the Zn sulfate or the Zn proteinate with moderate chelation strength (Zn-Prot M) for 26 days. The results showed that the plasma Zn contents from the hepatic portal vein of broilers at 28 days and 39 days of age were increased (p < 0.05) by Zn addition and greater (p < 0.05) in the Zn-Prot M than in the Zn sulfate. On d 28, Zn addition upregulated (p < 0.05) mRNA expression of zinc transporter 1 (ZnT1), Zrt-irt-like protein 5 (ZIP5), y + L-type amino transporter 2 (y + LAT2) and b0,+-type amino acid transporter (rBAT), zinc transporter 4 (ZnT4) protein expression, and zinc transporter 9 (ZnT9) mRNA and protein expression in the duodenum. Moreover, ZnT9 mRNA expression, ZnT4, ZIP5, and rBAT protein expression, zinc transporter 7 (ZnT7), and y + LAT2 mRNA and protein expression in the duodenum of broilers on 28 days were higher (p < 0.05) in the Zn-Prot M than in the Zn sulfate. On d 39, supplemental Zn increased (p < 0.05) peptide-transporter 1 (PepT1) mRNA expression and y + LAT2 protein expression, while the mRNA expression of ZnT7 and Zrt-irt-like protein 3 (ZIP3) were higher (p < 0.05) for the Zn-Prot M than for the Zn sulfate in the duodenum. It was concluded that the Zn-Prot M enhanced the Zn absorption in the small intestine partially via upregulating the expression of ZnT4, ZnT7, ZnT9, ZIP3, ZIP5, y + LAT2, and rBAT in the duodenum of broilers.

The Effect of Bone Morphogenetic Protein 2 or Extracellular Signal-Regulated Kinase 1 Silencing on Phosphorus Utilization and Related Parameters in Primary Broiler Osteoblasts.

Two experiments were conducted to study the effect of bone morphogenetic protein 2 (BMP2) or extracellular signal-regulated kinase 1 (ERK1) silencing on phosphorus (P) utilization and related parameters in primary broiler osteoblasts. Experiment 1 was carried out to select the most efficacious siRNAs against BMP2 or ERK1 for the subsequent experiment. In experiment 2, with or without the siRNA against BMP2 or ERK1, primary broiler osteoblasts were incubated in the medium supplemented with 0.0 or 2.0 mmol/L of P as NaH2PO4 for 12 days. The osteoblastic P utilization and related parameters were determined. The results showed that the si980 and si1003 were the most effective (P < 0.05) in inhibiting BMP2 and ERK1 expressions, respectively. The BMP2 silencing reduced (P < 0.004) the osteoblastic P retention rate, alkaline phosphatase (ALP) activity, BMP2 mRNA and protein expressions. Supplemental P increased (P = 0.0008) ALP activity. Significant interactions (P < 0.04) between the gene silencing and supplemental P level were observed in both mineralization formation and bone gal protein (BGP) content. The BMP2 silencing decreased (P < 0.05) mineralization formation at both 0.0 and 2.0 mmol/L of added P levels, but the decreased degree was greater at 2.0 mmol/L of added P level, while BMP2 silencing reduced (P < 0.05) BGP content at only 2.0 mmol/L of added P level. The ERK1 silencing decreased (P < 0.004) mineralization formation, ALP activity, BGP content, ERK1 mRNA, ERK1 and p-ERK1 protein expressions. Supplemental P increased (P < 0.03) mineralization formation, ALP activity, BGP content and p-ERK1 protein expression, but inhibited (P = 0.014) ERK1 protein expression. There was an interaction (P < 0.03) between the gene silencing and supplemental P level in the osteoblastic P retention rate. The ERK1 silencing decreased (P < 0.05) it regardless of 0.0 or 2.0 mmol/L of added P level, but the reduced degree was greater at 2.0 mmol/L of added P level. It was concluded that either BMP2 or ERK1 silencing suppressed P utilization, and thus either of them participated in regulating P utilization in primary broiler osteoblasts.

High-throughput UPLC-Q-TOF-MS/MS coupled with multivariable data processing approach for the rapid screening and characterization of chemical constituents and potential bioactive compounds from Danggui Shaoyao San.

Danggui Shaoyao San (DSS), a herbal formula, has been widely used for decades in China to treat senile dementia and dysmenorrhea. Here, an integrative high-throughput UPLC-Q-TOF-MS/MS method coupled with a multivariable data processing approach was established for rapidly screening and identifying chemical constituents and potential bioactive compounds from DSS. Through the comparison with mass fragment ions, relevant literature, and in-house reference material database coupled with MS cleavage mechanism, 150 chemical constituents, mainly including triterpenoids, flavonoids, phathalides, and organic acids, were tentatively characterized. Most of them were identified for the first time. Then, principal component analysis was used to evaluate the differences in chemical profiles between groups, whereas the variable importance of the projection (VIP) spectrum (VIP > 1) and the trend plot of orthogonal partial least squares discriminant analysis were applied to intuitively screen the candidate variables present only in the dosed group. Consequently, by comparison with all the characterized components in vitro, 23 potential bioactive compounds were successfully identified, comprising 5 triterpenoids, 4 phathalides, 4 flavonoids, 4 organic acids, 3 lactones, and 3 other compounds, which were present in various medicinal materials, reflecting a synergistic mechanism. This work developed a rapid, reliable, and robust approach for comprehensive characterization of the chemical components and potential bioactive compounds of DSS, providing solid data for further research on pharmacodynamic substances and pharmacological mechanisms of DSS.

Dentin Matrix Protein 1 Silencing Inhibits Phosphorus Utilization in Primary Cultured Tibial Osteoblasts of Broiler Chicks.

Three experiments were carried out in the present study to investigate whether dentin matrix protein 1 (DMP1) was involved in regulating phosphorus (P) metabolic utilization in primary cultured tibial osteoblasts of broiler chicks. Experiment 1 was conducted to select the optimal osteogenic inductive culture medium and the optimal induction time in primary cultured tibial osteoblasts of broiler chicks. In experiment 2, the siRNAs against DMP1 were designed, synthesized and transfected into primary cultured tibial osteoblasts of broiler chicks, and then the inhibitory efficiencies of siRNAs against DMP1 were determined, and the most efficacious siRNA was selected to be used for the DMP1 silencing. In experiment 3, with or without siRNA against DMP1, primary cultured tibial osteoblasts of broiler chicks were treated with the medium supplemented with 0.0, 1.0 or 2.0 mmol/L of P as NaH2PO4 for 12 days. The P metabolic utilization-related parameters were measured. The results showed that the osteogenic induced medium 2 and 12 days of the optimal induction time were selected; Among the designed siRNAs, the si340 was the most effective (P < 0.05) in inhibiting the DMP1 expression; DMP1 silencing decreased (P < 0.05) the expressions of DMP1 mRNA and protein, P retention rate, mineralization formation, alkaline phosphatase activity and bone gla-protein content in tibial osteoblasts at all of added P levels. It is concluded that DMP1 silencing inhibited P utilization, and thus DMP1 was involved in regulating P metabolic utilization in primary cultured tibial osteoblasts of broiler chicks, which provides a novel insight into the regulation of the P utilization in the bone of broilers, and will contribute to develop feasible strategies to improve the bone P utilization efficiency of broilers so as to decrease its excretion.

Dietary fiber modulates abdominal fat deposition associated with cecal microbiota and metabolites in yellow chickens.

Excessive deposition of abdominal fat is a public concern in the yellow chicken industry related to human nutrition. The common practice of nutritionists is to increase the fiber content in feed to control abdominal fat deposition of chickens. Corncob meal (CCM) is the cheapest ingredient widely used in animal diets. The possible effects of CCM on chicken abdominal fat deposition and the possible mechanism involving cecal microbiota remain unknown. The objectives of this study were to investigate the effects of CCM in modulating abdominal fat deposition and the role of the cecal microbiota and their metabolites. A total of 200 ninety-day-old Huxu female chickens were divided into 2 dietary treatments, each with 10 replicates of 10 birds, and were fed two finisher diets, from 90 to 135 d. The diets were a typical corn-soybean control diet (CON) and that diet with CCM partially replacing corn and corn gluten meal. Results showed that the CCM diet markedly decreased live weight and abdominal fat percentage (P < 0.05); chickens fed the CCM diet exhibited lower (P < 0.01) expression in abdominal fat of fatty acid binding protein 4 (FABP4), stearoyl-CoA desaturase (SCD), fatty acid synthase (FAS), and peroxisome proliferator-activated receptor γ (PPARγ) but higher (P < 0.05) expression of estrogen receptor alpha (ESR1). The CCM increased the abundance of Akkermansia (P < 0.05) and markedly reduced the relative cecal abundance of Phascolarctobacterium (P < 0.01), Rikenellaceae (P < 0.05), and Faecalibacterium (P < 0.01). The metabolomic and biochemical analyses demonstrated that the CCM diet increased (P < 0.05) the concentrations of butyrate in cecal contents. The majority of the metabolites in cecal digesta with differences in abundance were organic acids. The CCM diet increased (P < 0.05) contents of (R)-5-diphosphomevalote, pantothenic acid, 2-epi-5-epi-valiolone 7-phosphate, D-ribose 5-diphosphate, arbutin 6-phosphate, D-ribitol 5-phosphate, undecanoic acid, nicotinic acid, 4-methyl-2-oxovaleric acid, while decreasing (P < 0.05) those of oleic acid, glutaric acid, adipic acid, suberic acid, and L-fuculose 1-phosphate. In conclusion, these findings demonstrated that the dietary CCM treatment significantly decreased abdominal fat and altered the cecal microbiota and metabolite profiles of the yellow chickens.

Activity Guided Isolation of Phenolic Compositions from Anneslea fragrans Wall. and Their Cytoprotective Effect against Hydrogen Peroxide Induced Oxidative Stress in HepG2 Cells.

Anneslea fragrans Wall., commonly known as "Pangpo Tea", is traditionally used as a folk medicine and healthy tea for the treatment of liver and intestine diseases. The aim of this study was to purify the antioxidative and cytoprotective polyphenols from A. fragrans leaves. After fractionation with polar and nonpolar organic solvents, the fractions of aqueous ethanol extract were evaluated for their total phenolic (TPC) and flavonoid contents (TFC) and antioxidant activities (DPPH, ABTS, and FRAP assays). The n-butanol fraction (BF) showed the highest TPC and TFC with the strongest antioxidant activity. The bio-guided chromatography of BF led to the purification of six flavonoids (1-6) and one benzoquinolethanoid (7). The structures of these compounds were determined by NMR and MS techniques. Compound 6 had the strongest antioxidant capacity, which was followed by 5 and 2. The protective effect of the isolated compounds on hydrogen peroxide (H2O2)-induced oxidative stress in HepG2 cells revealed that the compounds 5 and 6 exhibited better protective effects by inhibiting ROS productions, having no significant difference with vitamin C (p > 0.05), whereas 6 showed the best anti-apoptosis activity. The results suggest that A. fragrans could serve as a valuable antioxidant phytochemical source for developing functional food and health nutraceutical products.

High dietary copper induces oxidative stress and leads to decreased egg quality and reproductive performance of Chinese Yellow broiler breeder hens.

The objective of this study was to investigate the effects of dietary copper (Cu) on production, egg quality, and hatchability of Chinese Yellow broiler breeder hens and growth performance of their offspring. A total of 576 30-week-old hens were randomly allotted into 6 groups, each with 6 replicates (8 cages for each replicate with 2 birds per cage). The basal diet contained 3.50 mg/kg Cu, and the other 5 treatment diets contained 8.5, 13.5, 23.5 43.5, and 83.5 mg/kg Cu, respectively, additionally supplemented with Cu on the basal diet. The trial lasted for 15 wk. Qualified egg rate of birds fed 23.5 or 83.5 mg/kg Cu was significantly decreased (P < 0.05) compared with those given 3.5, 8.5, or 13.5 mg/kg Cu. Plasma malondialdehyde concentration showed quadratic effect (P = 0.002) which that decreased first then increased with dietary Cu increased. Highest values of Cu content and hepatic activity of Cu-ATPase occurred in hens fed 83.5 mg/kg dietary Cu with linear (P = 0.001) and quadratic (P = 0.001) effects. Shell strength and proportion on 18th day of live embryos of hens fed 13.5 mg/kg Cu were the greatest compared with other groups respectively (P < 0.05); rate of qualified eggs for hatch and hatchability of fertilized eggs of hens fed 83.5 mg/kg Cu were the least (P < 0.05). In conclusion, both inadequate (3.5 mg/kg diet) and excess (83.5 mg/kg) of dietary Cu can induce oxidative stress in hens and lead to decreased egg quality. Hatchability and growth performance of offspring were decreased when breeder hens were fed excess Cu in spite of greater hatching weight. The appropriate dietary Cu level for Chinese Yellow broiler breeder hens during the egg-laying period is 15.7 to 21.2 mg/kg (1.81-2.44 mg Cu fed per day) when based on Cu level and Cu-ATPase activity in the liver. This dietary Cu requirement is approximately doubled (∼40 mg/kg, ∼4.60 mg Cu per bird per day) for maximal response of eggshell thickness.

Humic acid functionalized hyperbranched polytriazine based dispersive solid-phase extraction for acaricides determination in tea matrix.

Hyperbranched polytriazine functionalized with humic acid was prepared and developed as new sorbents for dispersive solid-phase extraction of three acaricides (clofentezine, fenpyroximate, and pyridaben) in tea samples combined with high-performance liquid chromatography detection. The sorbents were characterized by scanning electron microscopy, energy dispersive spectroscopy, Zeta-potential, and Fourier transform infrared spectroscopy. The extraction parameters (extraction time, ionic strength, desorption conditions) were optimized. The adsorption mechanism was evaluated utilizing Fourier transform infrared spectra. Under optimum conditions, satisfactory analytical performances were achieved, which included high precision (1.33-9.62%), low limits of detection (0.19-3.54 µg/L), and wide linear range (2.5-500 µg/L) for the analysis of the acaricides. Moreover, the proposed method proved highly effective for the determination of acaricides in tea samples, with the relative recoveries in the range of 65.20-108.13% and relative standard deviations < 9.87%. The method has great application potential for the detection of acaricides in tea samples.

Effects of dietary perilla seed oil supplementation on lipid metabolism, meat quality, and fatty acid profiles in Yellow-feathered chickens.

This study evaluated the effect of the dietary replacement of 1% lard (CT) with 1% perilla oil (PO), 0.9% perilla oil + 0.1% anise oil (PA), or 0.9% perilla oil + 0.1% ginger oil (PG) on indices of lipid metabolism, antioxidant capacity, meat quality, and fatty acid profiles from Yellow-feathered chickens at day 63. Compared with the CT chickens, those given perilla oil had decreased (P < 0.05) plasma lipid levels including triglycerides (TG), total cholesterol (TCH), and low-density lipoprotein cholesterol (LDL-C). Hepatic TG, TCH levels, and fatty acid synthase activity were also decreased (P < 0.05) in chickens fed diets containing perilla oil. Abdominal fat percentage was significantly decreased in birds fed the PG compared to CT diets. Birds fed the PA or PG diets had increased (P < 0.05) hepatic total SOD, glutathione peroxidase, and glutathione-S-transferase than in chickens given PO alone. In addition, the content of reduced glutathione (GSH) in breast muscle was lower (P < 0.05) in birds fed PO compared with those given PG, and the reverse was true for content of malondialdehyde. Compared with the CT diet, the PO diet decreased breast muscle shear values and increased yellowness (b*) of breast muscle (P < 0.05). Birds fed the PA or PG diets had meat with better overall acceptability than those fed the CT diet. Chickens fed perilla oil diets exhibited higher contents of α-linolenic acid (C18:3n-3), DHA (22:6n-3), polyunsaturated fatty acids, and n-3 fatty acids, together with a lower content of myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), total saturated fatty acids, and n-6/n-3 ratio compared to controls (P < 0.05). These findings indicate that perilla oil has the potential to decrease lipid-related indices and improve fatty acid profiles of breast meat in chickens without adverse effect on antioxidant status or meat quality; this was even better when perilla oil was given together with anise oil or ginger oil.