Cuscuta chinensis flavonoids alleviate ovarian damage in offspring female mice induced by BPA exposure during pregnancy by regulating the central carbon metabolism pathway.

Pregnancy is a sensitive window period for bisphenol A (BPA) exposure. BPA can pass through the placenta and cause reproductive damage in offspring female mice. Even BPA that is not metabolized during lactation can be passed through milk. Cuscuta chinensis flavonoids (CCFs) can alleviate reproductive damage caused by BPA, but the mechanism of action is unclear. To investigate the potential mitigating impact of CCFs on ovarian damage resulting from BPA exposure during pregnancy, we administered BPA and CCFs to pregnant mice during the gestational period spanning from 0.5 to 17.5 days. Aseptic collection of serum and ovaries from female mice was conducted on postnatal day 21 (PND21). Serum hormone levels and tissue receptor levels were quantified utilizing ELISA and PCR, while ovaries underwent sequencing and analysis through transcriptomics and metabolomics techniques. Additionally, the assessment of ovarian oxidative stress levels was carried out as part of the comprehensive analysis. The results showed that CCFs administration mitigated the adverse effects induced by BPA exposure on ovarian index, hormone levels, receptor expression, and mRNA expression levels in female offspring mice. The joint analysis of transcriptome and metabolome revealed 48 enriched pathways in positive ion mode and 44 enriched pathways in negative ion mode. Among them, the central carbon metabolism pathway is significantly regulated by BPA and CCFs. The screened sequencing results were verified through qPCR and biochemical kits. In this study, CCFs may participate in the central carbon metabolism pathway by reducing the expression of Kit proto-oncogene (Kit), hexokinase 1 gene (Hk1) and pyruvate kinase M (Pkm) mRNA and increasing the expression of h-ras proto-oncogene (Hras), sirtuin 3 (Sirt3), sirtuin 6 (Sirt6) and TP53 induced glycolysis regulatory phosphatase gene (Tigar) mRNA, thereby resisting the effects of BPA on the body. At the same time, the metabolic levels of D-Fructose 1,6-bisphosphate and L-Asparagine tend to be stable. Moreover, CCFs demonstrated a capacity to diminish the BPA-induced escalation in reactive oxygen species (ROS) and malondialdehyde (MDA). Simultaneously, it exhibited the ability to elevate levels of glutathione (GSH) and catalase (CAT), thereby effectively preventing peroxidation. In summary, CCFs alleviate BPA-induced ovarian damage in offspring female mice by regulating the central carbon metabolism pathway. This study will improve the information on BPA reproductive damage antagonist drugs and provide a theoretical basis for protecting animal reproductive health.

Salvia miltiorrhiza polysaccharide mitigates AFB1-induced liver injury in rabbits.

Aflatoxin B1 (AFB1) is one of the common dietary contaminants worldwide, which can harm the liver of humans and animals. Salvia miltiorrhiza polysaccharide (SMP) is a natural plant-derived polysaccharide with numerous pharmacological activities, including hepatoprotective properties. The purpose of this study is to explore the intervention effect of SMP on AFB1-induced liver injury and its underlying mechanisms in rabbits. The rabbits were administered AFB1 (25 µg/kg/feed) and or treatment with SMP (300, 600, 900 mg/kg/feed) for 42 days. The results showed that SMP effectively alleviated the negative impact of AFB1 on rabbits' productivity by increasing average daily weight gain (ADG) and feed conversion rate (FCR). SMP reduced aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) levels in serum, ameliorating AFB1-induced hepatic pathological changes. Additionally, SMP enhanced superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) activity, and inhibited reactive oxygen species (ROS), malondialdehyde (MDA), 4-Hydroxynonenal (4-HNE), interleukin-1ß (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression, thus mitigating AFB1-induced oxidative stress and inflammatory responses. Moreover, SMP upregulated the expression of nuclear factor E2 related factor 2 (Nrf2), heme oxygenase 1 (HO-1), NADPH quinone oxidoreductase 1 (NQO1) and B-cell lymphoma 2 (Bcl2) while downregulating kelch like ECH associated protein 1 (Keap1), cytochrome c (cyt.c), caspase9, caspase3, and Bcl-2-associated X protein (Bax) expression, thereby inhibiting AFB1-induced hepatocyte apoptosis. Consequently, our findings conclude that SMP can mitigate AFB1-induced liver damage by activating the Nrf2/HO-1 pathway and inhibiting mitochondria-dependent apoptotic pathway in rabbits.

The Structural Characterization of a Polysaccharide from the Dried Root of Salvia miltiorrhiza and Its Use as a Vaccine Adjuvant to Induce Humoral and Cellular Immune Responses.

In order to supplement the research gap concerning Salvia miltiorrhiza polysaccharide extracted from Danshen in NMR analysis, and to clarify its immune enhancement effect as an adjuvant, we isolated and purified SMPD-2, which is composed of nine monosaccharides such as Ara, Gal, and Glc from Danshen. Its weight average molecular weight was 37.30 ± 0.096 KDa. The main chain was mainly composed of →4)-α-D-Galp-(1→, →3,6)-ß-D-Glcp-(1→ and a small amount of α-L-Araf-(1→. After the subcutaneous injection of SMPD-2 as an adjuvant to OVA in mice, we found that it enhanced the immune response by activating DCs from lymph nodes, increasing OVA-specific antibody secretion, stimulating spleen lymphocyte activation, and showing good biosafety. In conclusion, SMPD-2 could be a promising candidate for an adjuvant.

Aflatoxin B1 triggers apoptosis in rabbit hepatocytes via mediating oxidative stress and switching on the mitochondrial apoptosis pathway.

Aflatoxin B1 (AFB1) is considered the most toxic carcinogenic compound, and exposure to AFB1 is highly associated with hepatocellular carcinoma. The aim of this study was to investigate the effects of different doses of AFB1 on growth performance and the liver of rabbits, as well as explore its underlying mechanisms. A total of eighty 30-day-old meat rabbits were randomly divided into four treatments. The control group was fed a pollution-free diet, while the AFL, AFM, and AFH groups were fed contaminated diets containing 13 µg/kg, 19 µg/kg, and 25 µg/kg of AFB1, respectively. The results showed that AFB1 had detrimental effects on the production performance of rabbits, resulting in decreased weight gain. Additionally, AFB1 exposure was associated with increased activity of Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT), as well as decreased levels of total protein (TP) and albumin (ALB) in the serum. AFB1 induced the production of reactive oxygen species (ROS) and malondialdehyde (MDA) while inhibiting the activity of glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) activity in liver tissues. AFB1 decreased the mRNA transcription and protein expression of nuclear factor-erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NAD(P)H dehydrogenase quinone-1 (NQO-1). AFB1 not only decreased the contents of cytochrome P4501A2 (CYP1A2), cytochrome P4502A6 (CYP2A6) and cytochrome P4503A4 (CYP3A4) but also increased the content of AFB1-DNA adducts in the liver. Furthermore, AFB1 enhanced the expression of cytochrome c (cyt-c), caspase-9, caspase-3, and Bcl-2-associated X protein (Bax), while inhibiting the expression of B-cell lymphoma 2 (Bcl-2). Therefore, we demonstrated that AFB1 triggered apoptosis in rabbit hepatocytes via mediating oxidative stress and switching on the mitochondrial apoptosis pathway, and decreased rabbit performance.

Cuscuta chinensis flavonoids reducing oxidative stress of the improve sperm damage in bisphenol A exposed mice offspring.

Bisphenol A (BPA) is a common environmental endocrine disruptor, and overexposure is a threat to male reproduction. Although studies have confirmed that BPA exposure causes a decrease in sperm quality in offspring, the dosage used, and the underlying mechanism is not clear. The purpose of this study is to investigate whether Cuscuta chinensis flavonoids (CCFs) can antagonize or alleviate BPA-induced reproductive injury by analyzing the processes associated with BPA's impairment of sperm quality. BPA and 40 mg/kg bw/day of CCFs were administered to the dams at gestation day (GD) 0.5-17.5. Testicles and serum of male mice are collected on postnatal day 56 (PND56), and spermatozoa are collected to detect relevant indicators. Our results showed that compared with the BPA group, CCFs could significantly increase the serum contents of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone (T) in males at PND 56, as well as the transcription levels of estrogen receptor alpha (ERα), steroidogenic acute regulatory protein (StAR) and Cytochrome P450 family 11, subfamily A, and member 1 (CYP11A1). CCFs also significantly inhibit the production of reactive oxygen species (ROS), reduce oxidative stress, increase mitochondrial membrane potential, and reduce sperm apoptosis. It also has a certain regulatory effect on sperm telomere length and mitochondrial DNA copy number. These results suggest that CCFs can increase reproductive hormone and receptor levels in adult males by regulating the expression of oxidative stress correlated factors, and ultimately mitigate the negative effects of BPA on sperm quality in male mice.

Salvia miltiorrhiza polysaccharides alleviate florfenicol-induced inflammation and oxidative stress in chick livers by regulating phagosome signaling pathway.

Florfenicol (FFC) is a commonly used antibiotic in animal breeding, especially in broiler breeding. Previous studies found that FFC could affect the liver function of chickens. However, the mechanisms underlying the effects of FFC on liver function are still not completely clear. Moreover, the research on drugs that antagonize FFC hepatotoxicity is relatively lacking. Salvia miltiorrhiza polysaccharides (SMPs) have been proved to have obvious liver protection effects. Therefore, we exposed chicks to FFC at the clinically recommended dose of 0.15 g/L. At the same time, 0.15 g/L FFC and 5 g/L SMPs were given to another group of chicks. After 5 days of continuous administration, the livers of chicks from different treatment groups were sequenced by transcriptome and proteome. Based on the analysis of sequencing data, we also focused on the detection of inflammation and oxidation indicators related to the phagosome signaling pathway with significant enrichment of differential factors in the livers of chicks. The results showed that some significantly differentially expressed genes and proteins induced by FFC were enriched in the phagosome signaling pathway, and they increased the expression levels of inflammatory factors and peroxides. However, SMPs intervention significantly reversed the tendency of FFC to alter phagosome signaling pathways and reduced the expression levels of inflammatory factors and peroxides. In conclusion, FFC caused liver inflammation and oxidative stress in chicks by regulating the phagosome signaling pathway. Meanwhile, SMPs could improve the adverse effects of FFC on the phagosome signaling pathway. This study provided new insights into the ameliorative effects and mechanisms of SMPs on hepatotoxicity of FFC.

Transcriptome and Metabolome Analyses Reveal Perfluorooctanoic Acid-Induced Kidney Injury by Interfering with PPAR Signaling Pathway.

Perfluorooctanoic acid (PFOA) is widely used in aviation science and technology, transportation, electronics, kitchenware, and other household products. It is stable in the environment and has potential nephrotoxicity. To investigate the effect of PFOA exposure during pregnancy on the kidneys of offspring mice, a total of 20 mice at day 0 of gestation were randomly divided into two groups (10 mice in each group), and each group was administered 0.2 mL of PFOA at a dose of 3.5 mg/kg or deionized water by gavage during gestation. The kidney weight, kidney index, histopathological observation, serum biochemistry, transcriptomics, and metabolomics of the kidneys of the 35-day offspring mice were analyzed. In addition, malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) levels in the kidney were measured. Transcriptome analysis results showed that 387 genes were up-regulated and 283 genes were down-regulated compared with the control group. These differentially expressed genes (DEGs) were mainly concentrated in the peroxisome-proliferator-activated receptor (PPAR) signaling pathway and circadian rhythm. Compared with the control group, 64 and 73 metabolites were up- and down-regulated, respectively, in the PFOA group. The altered metabolites were mainly enriched in the biosynthesis of unsaturated fatty acids. PFOA can affect the expression levels of circadian rhythm-related genes in the kidneys of offspring mice, and this change is influenced by the PPAR signaling pathway. PFOA causes oxidative stress in the kidneys, which is responsible for significant changes in metabolites associated with the biosynthesis of unsaturated fatty acids.

Salvia miltiorrhiza polysaccharides ameliorates Staphylococcus aureus-induced mastitis in rats by inhibiting activation of the NF-κB and MAPK signaling pathways.

The lactation capacity of dairy cows is critical to the productivity of the animals. Mastitis is a disease that directly affects the lactation capacity of cows. Staphylococcus aureus (S. aureus) is one of the most important pathogens that causes mastitis in dairy cows. The anti-inflammatory effect of Salvia miltiorrhiza polysaccharides (SMPs) has been demonstrated in mice and chickens. However, the effectiveness of SMPs in preventing and treating mastitis is unclear. Therefore, the purpose of this study was to explore the protective effect and mechanism of SMPs on mastitis caused by S. aureus. S. aureus was used to induce mastitis in rats, and three doses of SMPs (87.5, 175, 350 mg/kg, BW/d) were administered as treatments. The bacterial load, histopathology, and myeloperoxidase (MPO) and N-acetyl-ß-D-glucosaminidase (NAGase) activities of mammary glands were observed and measured. Cytokines, including interleukin (IL)-1ß, interleukin (IL)-6, and tumor necrosis factor α (TNF-α), were examined by qRT-PCR and ELISA. Key proteins in the NF-κB and MAPK signaling pathways were analyzed by Western blotting. The results showed that SMP supplementation could significantly reduce the colonization of S. aureus and the recruitment of inflammatory cells in mammary glands. S. aureus-induced gene transcription and protein expression of IL-1ß, IL-6, and TNF-α were significantly suppressed in mammary glands. In addition, the increase in NF-κB and MAPK protein phosphorylation was inhibited by SMPs. These results revealed that supplementation with SMPs protected the mammary gland of rats against damage caused by S. aureus and alleviated the inflammatory response. This study provides a certain experimental basis for the treatment of S. aureus-induced mastitis with SMPs in the future.

Transcriptomics and metabolomics revealed the molecular mechanism of the toxic effect of mancozeb on liver of mice.

Mancozeb (MCZ), a broad-spectrum fungicide, has been widely used in crops (tomatoes and potatoes) in the past few decades, resulting in its bioaccumulation in the food web. However, the mechanism of MCZ on liver injury has not been reported yet. This study combined transcriptomics and metabolomics to explore the potential mechanism of MCZ on liver injury. MCZ group was given 100 mg/kg MCZ every day, and the C group was given 0.2 mL of deionized water every day. One hundred mg/kg MCZ led to unclear hepatocyte structure and hemorrhagic inflammatory cell infiltration. Transcriptomics and metabolomics analyses showed that the MCZ group resulted in 326 differentially expressed genes (DEGs) and 179 differential metabolites. Joint analysis showed that DEGs and differential metabolites were mainly enriched in the adenosine monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway. We found that MCZ could increase the content of reactive oxygen species (ROS) and reduce the activities of superoxide dismutase (SOD) and catalase (CAT). The contents of DNA methyltransferases (DNMT1, DNMT3A, and DNMT3B) in the liver decreased significantly, and the state of DNA methylation was significantly higher than the control (C) group (p < 0.05). Our results suggest that AMPK and mitogen­activated protein kinase (MAPK) signaling pathways play an important role in MCZ-induced liver injury and are the key mechanisms for understanding the hepatotoxicity of MCZ.

Effects of early florfenicol exposure on glutathione signaling pathway and PPAR signaling pathway in chick liver.

Florfenicol (FFC) is a common antibiotic for animals. The nonstandard and excessive use of FFC can cause veterinary drug residues in animals, pollute soil and marine environment, and even threaten human health. Therefore, it is necessary to study the toxicity and side effects of FFC on animals. Our previous studies have proved that FFC can cause liver injury in chicks, but there are few in-depth studies on the mechanism of FFC causing liver injury at the level of signaling pathway in chicks. Therefore, transcriptome and proteome sequencing were performed and combined analysis was performed. Sequencing results showed that 1989 genes and 917 proteins were significantly changed in chick livers after FFC exposure. These genes and proteins are related to redox, glutathione transferase activity and lipid metabolism. There are 9 significantly different genes and 7 significantly different proteins in glutathione signaling pathway. Oxidative stress may occur in the liver of chicks through the change of activation state of glutathione signaling pathway. And there are 13 significantly different genes and 18 significantly different proteins in PPAR signaling pathway. The changes of PPAR signaling pathway may induce lipid metabolism disorder in liver. The verification results of qPCR and PRM were consistent with the sequencing results. We also detected GSH-Px, GSH, GST, TG, TC and ANDP levels in liver. These changes of biochemical indicators directly confirmed oxidative stress and lipid metabolism disorders were occurred in the livers of chicks treated by FFC. In conclusion, FFC could induce liver injury in chicks by regulating the expression levels of significantly different genes and proteins in glutathione signaling pathway and PPAR signaling pathway.

Salvia miltiorrhiza polysaccharides alleviates florfenicol induced kidney injury in chicks via inhibiting oxidative stress and apoptosis.

Florfenicol (FFC) is a commonly used antibiotic in animal husbandry, which is easy to cause organs damage in a variety of animals. It has been proved to have nephrotoxicity and affect the yield and quality of meat products. Salvia miltiorrhiza polysaccharides (SMPs) have been proved to have the pharmacological effects of regulating immunity and protecting the liver of animals, and its alleviative effect on renal injury is unclear. In order to investigate the alleviating effect of SMPs on drug nephrotoxicity and determine its potential molecular mechanism, we took chicks as the research object, FFC as the induced drug, and established the model by adding SMPs in drinking water. The chicks were randomly divided into control group, FFC model group (0.15 g/L FFC), FFC + low, medium and high dose of SMPs groups (0.15 g/L FFC + 1.25, 2.5, 5 g/L SMPs) and SMPs group (5 g/L SMPs). The results showed that, SMPs increased the average weight gain and renal index of chicks, alleviated the pathological changes of renal structure induced by FFC, decreased the contents of uric acid, blood urea nitrogen and creatinine in serum and malondialdehyde in renal tissue, increased the levels of glutathione, superoxide dismutase and catalase in renal tissue, up-regulated the relative expression levels of nuclear factor erythroid 2 related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and nicotinamide adenine dinucleotide phosphate: quinone oxidoreductase-1 (NQO-1) mRNA and protein, and down-regulated the relative expression levels of p53, Caspase-3 and Caspase-6 mRNA and protein and the apoptosis rate of renal histiocytes. It is concluded that SMPs could significantly alleviate the renal injury induced by FFC, and its mechanism may be related to improving renal antioxidant capacity and inhibiting abnormal apoptosis of renal histiocytes.

Florfenicol causes excessive lipid peroxidation and apoptosis induced renal injury in broilers.

In order to study the effects and mechanism of florfenicol (FFC) on the kidney function of broilers, 180 1-day-old broilers were randomly divided into 6 groups, 30 in each group. Except for the control group, different doses of FFC were added to drinking water in the other 5 groups (0.15 g/L, 0.3 g/L, 0.6 g/L, 1.2 g/L and 1.8 g/L). After continuous administration for 5 days, renal histopathological changes, serum renal function indicators, renal peroxidation products and antioxidant factors, and apoptotic factors were detected in broilers aged 21 and 42 days. The results showed that compared with the control group, the kidney tissue structure was disordered, the glomerulus was atrophic, the cystic cavity was enlarged, and the epithelial cells of renal tubules were seriously vacuolated in broilers of treatment groups. And with the growth of broilers, the kidney injury of broilers in the low-dose FFC group was relieved. FFC significantly increased the contents of uric acid (UA), blood urea nitrogen (BUN), creatinine (CRE) in serum and malondialdehyde (MDA) in kidney of broilers, but significantly reduced the levels of glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) in kidney. FFC significantly inhibited the mRNA relative transcriptional levels of nuclear factor-erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and nicotinamide adenine dinucleotide phosphate: quinone oxidoreductase-1 (NQO-1), and increased the mRNA and protein expression levels of p53, Caspase-3 and Caspase-6 in kidney tissue of broilers. It is concluded that FFC has certain nephrotoxicity to broilers, and its effect on kidney is dose-dependent and reversible. FFC causes intense lipid peroxidation in broiler kidney by inhibiting the expression of related factors in the downstream signal pathway of Nrf2. FFC can also up-regulate the expression of pro-apoptotic factors and accelerate the abnormal apoptosis of renal cells, thus seriously affecting the renal function of broilers.

Synergistic use of florfenicol and Salvia miltiorrhiza polysaccharide can enhance immune responses in broilers.

To explore the effect of florfenicol (FFC) combined with Salvia miltiorrhiza polysaccharide (SMPs) on immune function of Broilers. One hundred and twenty-one-day-old chicks were chosen and divided into 6 groups. The group A received standard basal diet only, the group B received a basal diet with FFC (0.15 g/L diet), and the group C, D, E received a basal diet with FFC (0.15 g/L diet) and SMPs (1.25 g/L, 2.5 g/L, 5 g/L diet)，the group F received a basal diet with SMPs (5 g/L diet). FFC can significantly inhibit the growth performance of broilers, but has no significant damage to the immune function of broilers. The combination of FFC and SMPs can improve the growth performance of broilers, increase the number of leukocyte subtypes in blood (P < 0.05), increase the number of Newcastle disease (ND) and avian influenza (AI) antibodies in blood, the number of immunoglobulins, and the content of cytokines (P < 0.05). In addition, it significantly improve the lymphocyte conversion rate of broiler peripheral blood (P < 0.05). So that, synergistic use of FFC and SMPs can enhance immune responses in Broilers.

Florfenicol induces oxidative stress and hepatocyte apoptosis in broilers via Nrf2 pathway.

In order to explore the mechanism of liver injury induced by florfenicol (FFC) in broilers, one hundred and twenty broilers were randomly divided into six groups, twenty broilers in each group. Except for control group, the other five groups were given different doses of FFC (0.15 g/L, 0.3 g/L, 0.6 g/L, 1.2 g/L and 1.8 g/L) in drinking water. After five days of continuous use, blood was collected from the subpterional vein and the chickens' liver were obtained. Chicken weight gain and liver indices were calculated; blood routine analysis was performed; the oxidative stress and apoptosis of hepatocytes was detected. The results showed that compared with the control group, except for 0.15 g/L FFC, the other doses of FFC significantly decreased the weight gain, white blood cell (WBC) and platelet (PLT) contents in blood, 0.3 g/mL FFC and 1.8 g/L FFC significantly reduced the content of hemoglobin (RGB) (P < 0.05); all doses of FFC significant decreased red blood cell (RBC) increased Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) contents in serum of chickens (P < 0.05), and significantly decreased the contents of albumin (ALB) and total protein (TP) in serum (P < 0.05), but had no significant effect on alkaline phosphatase (ALP) contents(P > 0.05). FFC significantly increased malondialdehyde (MDA) content in serum and liver tissues, but decreased glutathione (GSH), Superoxide dismutase (SOD) and catalase (CAT) content (P < 0.05), and significantly inhibited the mRNA transcription and protein expression of antioxidant proteins nuclear factor-erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and NAD(P)H dehydrogenase quinone-1 (NQO-1)(P < 0.05). FFC also inhibited the content and the transcription level of cytochrome P4501A1(CYP1A1) and CYP2H1 in liver (P < 0.05). At the same time, FFC significantly promoted the apoptotic rate of hepatocytes and the mRNA transcription and protein expression of caspase-3 and caspase-6 (P < 0.05). With the increase of FFC concentration, liver injury became more and more serious, which affected liver function in chickens by inhibiting enzyme activity in Nrf2-ARE pathway to increase oxidative stress and promoting apoptotic protein expression to accelerate hepatocyte apoptosis.

Cuscuta chinensis flavonoids alleviate bisphenol A-induced apoptosis of testicular cells in male mice offspring.

Bisphenol A (BPA) is a widespread environmental endocrine disruptor that has multiple effects on reproductive organ development. To investigate the effect of Cuscuta chinensis flavonoids (CCFs) on testicular apoptosis induced by BPA in male mice offspring, pregnant mice were administered intragastrically with BPA and CCF at gestation day (GD) 0.5-17.5. The testes of male offspring (F1 males) were collected at post-natal day (PND) 21 and PND 56 for the detection of related indicators. The results showed that compared with the BPA group, the testicular index in CCF groups was significantly increased at PND 21 (p < .01). For the mice of different concentrations of CCF groups, the expression levels of bax, caspase-9 and caspase-7 proteins were significantly decreased at PND 21 and PND 56, while the expression level of bcl-2 protein was significantly increased, and testicular apoptotic cells were also decreased significantly (p < .01 or p < .05). Forty mg/kg CCF has no significant difference compared with the control group. The results indicated that CCF could protect the testis development of F1 male mice by alleviating the apoptosis of testicular cells induced by BPA.

Lonicera flos and Curcuma longa L. extracts improve growth performance, antioxidant capacity and immune response in broiler chickens.

Alternatives to antibiotics are urgently needed to maintain broiler growth and health. The present study was conducted to evaluate the effects of Lonicera flos and Curcuma longa L. extracts (LCE) as antibiotic substitutes on growth performance, antioxidant capacity and immune response in broilers. A total of 480 one-day-old female broilers (WOD168) were allocated to 3 treatments with 5 replicates of 32 birds for 35 days. The 3 treatments were: an antibiotic-free basal diet (control, CON), CON +50 mg/kg spectinomycin hydrochloride and 25 mg/kg lincomycin hydrochloride (ANT), CON +500 mg/kg LCE (LCE). During the entire experimental period, supplementation of ANT and LCE increased (p < 0.01) average daily gain (ADG) and decreased (p < 0.05) feed conversion ratio (FCR), thereby resulting in greater final body weight (BW) compared with CON. Dietary LCE supplementation increased (p < 0.05) serum (glutathione peroxidase) GSH-Px, (superoxide dismutase) SOD and total antioxidant capacity (T-AOC) activities, and decreased (p < 0.05) serum malonaldehyde (MDA) concentration at day 35 compared with CON. There was no significant difference in serum catalase (CAT) activity among treatments. Birds in LCE group had lower (p < 0.05) MDA concentration and higher SOD activity in liver than those in CON and ANT groups at day 35. Birds in LCE group had higher (p < 0.05) phagocytic index and serum antibody titers to Newcastle disease virus (NDV) than those in CON group. Lower (p < 0.05) concentrations of pro-inflammatory cytokines and higher (p < 0.05) concentrations of anti-inflammatory cytokines in serum and liver were observed in birds fed LCE diet than those fed CON diet. In conclusion, dietary supplementation of LCE improved growth performance by enhancing antioxidant capacity, strengthening immune system and alleviating inflammation, which has potential as antibiotic alternatives.

The effect of Lonicerae flos and Rhizoma curcumae longae extract on the intestinal development and function of broilers.

This study was conducted to explore effects of Lonicerae flos and Rhomoma curcumae longae extracts (LR) on intestinal function of broilers. Three hundred broiler chickens were randomly assigned to the following 5 groups. The control group were fed the basal diet; the antibiotic group were fed the basal diet supplemented with spectinomycin hydrochloride (50 million units/ton) + lincomycin hydrochloride (25 g/ton); the LRH, LRM and LRL groups were fed the basal diet supplemented with a high dose (750 g/ton of feed), normal dose (500 g/ton of feed), or low dose (250 g/ton of feed) of LR, respectively. The changes of intestinal structure, intestinal digestive enzyme activities, antioxidant enzyme activities, inflammatory cytokines, and bacterial abundances in the colon and cecum contents were determined. The results indicated that compared with the control group and the antibiotic group, LR significantly increased the villus length/crypt depth (VCR) of the intestine, and significantly inhibited oxidative stress and inflammatory responses in the broiler intestine. In addition, LR regulated intestinal function by increasing the abundance of the intestinal microorganisms in broilers. In conclusion, LR improved antioxidant capacity, intestinal morphology, and microorganisms, and inhibited inflammatory response. The effect of high and medium doses of LR was better than lower doses.

Biological Surface Layer Formation on Bioceramic Particles for Protein Adsorption.

In the biomedical fields of bone regenerative therapy, the immobilization of proteins on the bioceramic particles to maintain their highly ordered structures is significantly important. In this review, we comprehensively discussed the importance of the specific surface layer, which can be called "non-apatitic layer", affecting the immobilization of proteins on particles such as hydroxyapatite and amorphous silica. It was suggested that the water molecules and ions contained in the non-apatitic layer can determine and control the protein immobilization states. In amorphous silica particles, the direct interactions between proteins and silanol groups make it difficult to immobilize the proteins and maintain their highly ordered structures. Thus, the importance of the formation of a surface layer consisting of water molecules and ions (i.e., a non-apatitic layer) on the particle surfaces for immobilizing proteins and maintaining their highly ordered structures was suggested and described. In particular, chlorine-containing amorphous silica particles were also described, which can effectively form the surface layer of protein immobilization carriers. The design of the bio-interactive and bio-compatible surfaces for protein immobilization while maintaining the highly ordered structures will improve cell adhesion and tissue formation, thereby contributing to the construction of social infrastructures to support super-aged society.

Direct Immobilization of Folic Acid Molecules on Hydroxyapatite Nanoparticles with Substitution and Coordination Phenomena.

We successfully synthesized folic acid (FA) immobilized hydroxyapatite (HA) nanoparticles without using a mediative reagent (e.g., silane coupling agent), and the immobilization states were evaluated and discussed. The HA nanoparticles with higher biocompatibility have two different planes, namely, c- and m-planes. These plane surfaces are rich in phosphate groups (P-site) and Ca2+ ions (C-site), respectively. We suggested that during the synthesis of the HA nanoparticles, the P-site substitution and C-site coordination with the addition of organic molecules containing -COO- ions can occur. Thus, it is possible to simultaneously immobilize two molecules to one HA nanoparticle. In this study, we successfully synthesized FA-immobilized HA nanoparticles by P-site substitution and C-site coordination reactions, which were named as substitution type and coordination type. In the substitution type, when FA was reacted with HA during the nucleation stage, the PO43- ions of HA decreased as the FA ratio of coverage surface area increased, and the crystalline phase was changed significantly from the Ca deficient HA to the carbonated HA phase. Accordingly, it was indicated that FA was immobilized on HA by the P-site substitution. In the coordination type, since FA was reacted with HA after the completion of crystal growth, the crystalline phase was changed slightly as the FA ratio of coverage surface area increased, indicating that FA was immobilized on HA by the C-site coordination. From the above, we controlled the FA immobilization states on the HA nanoparticles by the P-site substitution and the C-site coordination through the FA addition timing in the synthesis. Since the -COO- ions in FA could be selectively substituted with the P-site in HA, it is possible to directly coordinate the foreign organic molecules to the Ca2+ ions in HA. Therefore, the immobilization technique of this study is expected to achieve two different drug molecules with diagnosis and therapy functions (i.e., theranostics) on one nanoparticle.

Effects of herbal dregs supplementation of Salvia miltiorrhiza and Isatidis Radix residues improved production performance and gut microbiota abundance in late-phase laying hens.

The present study was designed to evaluate the effect of a mixture of Chinese medicinal residues (CMRs) consisting of Salvia miltiorrhiza residues (SMR) and Isatidis Radix residues (IRR) on productive performance, egg quality, serum lipid and hormone levels, liver and blood antioxidant capacity, oviduct inflammation levels, and gut microbiota in the late-laying stage. A total of 288 fifty-four-week-old BaShang long-tailed hens were divided into four groups. The feed trial period was 8 weeks. The control group was fed the basic diet as a CCMR group, supplemented with 3, 4, and 6% for the experimental groups LCMR, MCMR, and HCMR. The egg production rate of the MCMR group was 8.1% higher than that of the CCMR group (p < 0.05). Serum triglyceride (TG) levels of hens of the CMR-supplemented group were significantly decreased than those of the CCMR group (p < 0.05). The group supplemented with different levels of CMR had significantly higher serum HDL-C levels compared with the control group (p < 0.05). Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were remarkably increased for the LCMR and MCMR groups and significantly decreased for the HCMR group compared to CCMR (p < 0.05). Serum and liver glutathione peroxidase (GSH-PX) activities were significantly increased, and malondialdehyde (MDA) levels were significantly decreased in the MCMR group compared to the CCMR group (p < 0.05). The expression levels of tubal inflammatory factor markers (IL-4, IL-1ß, TNF-α) in the MCMR and HCMR groups were consistent with the pathological findings of the sections. As for cecal microbiota, supplementation with CMR affected the alpha diversity of the cecum microbiome at the genus level. The Shannon index was significantly higher in the MCMR group than in the CCMR and HCMR groups (p < 0.05). Supplementation with different levels of CMR mainly regulated the ratio of intestinal Firmicutes to Bacteroidetes and the abundance of phyla such as Proteobacteria. In addition, CMR supplementation at different levels in the diet enriched lipid-metabolizing bacteria, such as Bacteroides and Ruminococcus_gnavus_group. Furthermore, according to linear discriminant analysis (LDA) effect size (LEfSe) analysis, the MCMR group showed an increase in the number of short-chain fatty acid-producing bacteria Romboutsia and fiber-degrading specialized bacteria Monoglobus. Therefore, supplementation of appropriate amounts of CMR to the diet of laying hens enhanced reproductive hormone levels, hepatic antioxidant capacity, and lipid metabolism, alleviated the levels of oviductal inflammatory factors, and modulated the abundance structure of bacterial flora to improve the late-laying performance and egg quality. The results of the current study showed that CMR is a beneficial feed supplement for chickens when added in moderation.