Effects of in ovo feeding and dietary addition oils on growth performance and immune function of broiler chickens.

This study aimed to investigate the effects of in ovo feeding (IOF) and dietary addition (DA) oils on growth, development and immune function of broiler chickens. In experiment 1, a total of 500 eggs were randomly assigned to 3 treatments: non-injected group (CON) with 100 eggs; soybean oil injected group (SO) with 200 eggs and linseed oil injected group (LO) with 200 eggs. Results showed that there were no detrimental effects of IOF of oils on embryonic development. In experiment 2, a two factor experimental design was adopted. After hatching, 120 chicks which came from each oil-injected group were divided into 2 treatments with 6 replicates, and chickens were fed soybean oil diet and linseed oil diet, respectively. The results showed that DA linseed oil increased final body weight (FBW) of broilers at d 21 post hatch, IOF of linseed oil decreased average daily feed intake (ADFI) and feed conversion ratio (FCR) of broilers from d 1 to 21 (P < 0.05), while the plasma leptin level of 21-day-old broilers was increased by IOF or DA linseed oil (P < 0.05). Main effect analysis showed that DA linseed oil increased the spleen index and mRNA expression of IFN-γ in spleen of broilers at 7 d of age (P < 0.05). IOF of linseed oil upregulated the mRNA expression of IFN-γ in the spleen of chicks at 1 d and mRNA expression of IL-2 and IL-4 in spleen of broilers at 21 d (P < 0.05), and the interaction effect showed that IOF and DA linseed oil synergically increased the expression of IL-2 and IL-4 in spleen of broilers at 21 d. Compared with SO group, LO increased the Shannon index of hatching-day cecum microflora (P < 0.05). Principal co-ordinates analysis (PcoA) showed that LO group clearly separated from CON and SO groups. Finally, Spearman correlation analysis also manifested that Alkalicoccus was significantly correlated with spleen index and mRNA expression of IL-2, and Phreatobacter was significantly correlated with the mRNA expression of IL-2 and IFN-γ in spleen, Acinetobacter had a positive correlation with thymus index (P < 0.05). In conclusion, IOF of linseed oil reduced the ADFI and FCR of broilers and increased the species diversity and changed the structure of cecal microflora of chicken embryos at the 19th day of incubation (E19). Immune function of broilers spleen was also regulated by IOF and DA linseed oil.

The ginger extract could improve diabetic retinopathy by inhibiting the expression of e/iNOS and G6PDH, apoptosis, inflammation, and angiogenesis.

Diabetic retinopathy is a complication of diabetes, caused by high blood sugar levels damaging the retina. It is the result of damage to the small blood vessels and neurons of the retina. Ginger and its phytochemical compounds can improve oxidative damage and inflammation. However, the effects of this plant on ocular expression G6PDH and e/iNOS, eye cell apoptosis, and angiogenesis are not well known in this tissue. Therefore, the aim of this study was to evaluate the therapeutic potential of ginger extract on rats with type 2 diabetic retinopathy. Thirty-two Wistar rats were randomly divided into four controlled and treated groups. The serum level of metabolic factors such as lipid profiles, insulin and glucose, and the level of oxidative biomarkers along with the TNF-α level in eye tissue were measured. The expression of NF-κB, VEGF, BAX, Bcl-2, caspase-3, e/iNOS, and G6PDH in eye tissue was measured. Serum levels of lipid profiles, glucose, and insulin, oxidative and inflammatory markers were significantly increased in the diabetic group compared to control. While, treatment with ginger extract could significantly improve these factors in diabetic rats. Moreover, the ocular expression of e/iNOS, G6PDH, VEGF, NF-κB, and genes involved in apoptosis was changed in diabetic rats. However, treatment with ginger extract could ameliorate these changes in the diabetic-treated group. It can be concluded that ginger extract could improve diabetic retinopathy by inhibiting oxidative damage, inflammation, iNOS, VEGF, apoptosis, and improving eNOS and G6PDH. PRACTICAL APPLICATIONS: Microvascular complications of diabetes such as retinopathy can be one of the main causes of disability in people with diabetes. Chronic hyperglycemia, oxidative stress, inflammation, and apoptosis cause diabetic retinopathy through retinal damage. Ginger, on the other hand, is an available, inexpensive, and uncomplicated medicinal plant that contains more than 20 different phytochemicals, such as gingerol and shogaol, which have anti-inflammatory, antioxidant, antihypertensive, hypoglycemic, and hypolipidemic properties. The results of our study showed well that the ginger extract could improve diabetic retinopathy by inhibiting the expression of e/iNOS and G6PDH and oxidative damage, apoptosis, inflammation, and angiogenesis. Therefore, ginger and its compounds can be a good option to improve the complications of diabetes.

Potato tillage method is associated with soil microbial communities, soil chemical properties, and potato yield.

Intensive potato continuous cropping (IPCC) results in low potato yields compared with non-intensive potato continuous cropping (PCC) and potato-maize rotation (PMRC). However, it is still unclear whether the degree of potato continuous cropping obstacle is related to the soil environment formed by the previous crop. To investigate the effect of planting potatoes and planting maize after harvesting the spring potatoes on soil chemical properties and soil microbial community structure, an experiment was carried out in the same origin soil environment over a period of seven years: (a) PCC, i.e., spring planting; (b) IPCC, i.e., autumn and spring planting (IPCC); (c) PMRC, i.e., spring potatoes and summer maize (PMRC), and (d) fallow (CK). We confirmed that the potato yield under PMRC was significantly higher than that under PCC and IPCC. Under IPCC, soil total phosphorus content was significantly higher than other treatments, whereas ammonium nitrogen content was the lowest. Compared with PCC and IPCC, PMRC had a higher ammonium nitrogen content and lower total phosphorus content. The significantly different fungal taxa in IPCC (Glomerellales, Plectosphaerella, Thelebolales) may threaten the health of the plant and positive correlated with soil total phosphorus, while other microbial taxa in PMRC (Bacillales, Polythrincium, Helotiales) can mainly promotes plant nitrogen uptake and protects plants against diseases. The PMRC-promoting taxa were positively correlated with the ammonium nitrogen content and negative correlated with soil total phosphorus content. In summary, the cropping systems might have affected potato yields by changed soil microorganism community structures - especially fungal community structures - and by the chemical properties of the soils that also depends on microbes.

Study on the interactive effect of deoxynivalenol and Clostridium perfringens on the jejunal health of broiler chickens.

With global warming and ban on antibiotics, it occurs occasionally that deoxynivalenol (DON) together with Clostridium perfringens impairs the gut health of broiler chickens. However, the interactive effect of DON and C. perfringens on intestinal health is still unknown. A total of 120 one-day-old Arbor Acres broilers were randomly distributed to 4 groups. Birds were gavaged with C. perfringens (8 × 108 CFU/d per bird) or sterile medium and fed a DON diet (0 or 5 mg of DON per kg diet) to investigate the interactive effects. The main effect analysis showed that DON diet significantly downregulated (P < 0.05) the mRNA expression of mucin-2, B-cell lymphoma-2-associated X, and cysteinyl aspartate-specific proteinase-3 of jejunal mucosa; decreased (P < 0.05) the indexes of ACE, Chao1, Shannon, and Simpson; and also decreased the relative abundance of the phylum Bacteroidete and the genera Lactococcus in jejunal contents of broilers chickens. Meanwhile, C. perfringens significantly increased (P < 0.05) crypt depth; decreased (P < 0.05) the ratio of villi height to crypt depth, the activity of jejunal diamine oxidase, and the relative abundance of Lactococcus; and upregulated (P < 0.05) the relative expression of B-cell lymphoma-2 and cysteinyl aspartate-specific proteinase-8. Furthermore, the interactions between DON and C. perfringens were most significant (P < 0.05) in the mRNA expression of lipopolysaccharide-induced TNF factor (LITAF) and TLR-4, the abundance of the genera Lactococcus in jejunal contents, and butyric acid concentrations in cecal contents of birds. Finally, Spearman correlation analysis suggested that the most negative correlations (P < 0.05) with the abundance of the genera except Lactobacillus were observed within the mRNA expression of LITAF. The abundance of Lactococcus had a positive correlation (P < 0.05) with the expression of Caspase-3. Most genera except Lactobacillus negatively correlated (P < 0.05) with acetic acid, butyric acid, and total short-chain fatty acids. In conclusion, dietary deoxynivalenol and C. perfringens challenge had a harmful effect on the jejunal health and should be carefully monitored in broiler production.

Accurate discrimination of Gastrodia elata from different geographical origins using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis.

Gastrodia elata from different geographical origins varies in quality and pharmacological activity. This study focused on the classification and identification of Gastrodia elata from six producing areas using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis. Before recognition analysis, a principal component analysis was applied to ascertain the discrimination possibility with high-performance liquid chromatography fingerprints. And then, boosting partial least-squares discriminant analysis and conventional partial least-squares discriminant analysis were applied in this study. Experimental results indicated that the adaptive iteratively reweighted penalized least-squares algorithm could eliminate the baseline drift of high-performance liquid chromatography chromatograms effectively. And compared with partial least-squares discriminant analysis, the total recognition rates using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis for the calibration sets and prediction sets were improved from 94 to 100% and 86 to 97%, respectively. In conclusion, high-performance liquid chromatography combined with boosting partial least-squares discriminant analysis, which has such advantages as effective, specific, accurate, non-polluting, has an edge for discrimination of traditional Chinese medicine from different geographical origins. And the proposed methodology is a useful tool to classify and identify Gastrodia elata from different geographical origins.

Identification of amentoflavone as a potent highly selective PARP-1 inhibitor and its potentiation on carboplatin in human non-small cell lung cancer.

BACKGROUND: Nuclear protein poly (ADP-ribose) polymerase-1 (PARP-1) is a key enzyme in the repair of DNA and is a promising target in the development of chemosensitizers. This study first investigated the inhibitory effects of amentoflavone (AMF) and its derivatives on PARP-1 and the potentiation of AMF on carboplatin (CBP) in non-small cell lung cancer (NSCLC). PURPOSE: This study aims to evaluate the inhibitory effect of AMF against PARP-1 and its potentiation on CBP in lung cancer both in vitro and in vivo. STUDY DESIGN: The inhibitory effect of AMF on PARP-1 was investigated using molecular docking and cell-free model of PARP-1 assay. Its potentiation on CBP in lung cancer was also evaluated. METHODS: Fluorescence resonance energy transfer assay was used to detect the inhibitory effects of AMF and its analogues on PARP-1. Molecular docking was employed to predict the binding mode of AMF and PARP-1. MTT assay, isobologram analysis, Hoechst staining, and Annexin V-PI double staining were used to confirm the potentiation of AMF on CBP in vitro. siRNA (PARP-1)-A549 cells were used to reveal the action target of AMF. Western blot analysis, immunohistochemistry, and Tunnel assay were employed to evaluate the potentiation of AMF on CBP in A549 xenograft mice. RESULTS: AMF and its analogues exerted excellent inhibitory effects on PARP-1 with IC50 values ranging from 0.198  µM to 0.409  µM. Docking experiment showed that AMF can stably bind to PARP-1 with a comparable binding energy to olaparib. AMF can decrease the expression of PAR induced by H2O2in vitro. AMF synergistically increased the CBP anti-proliferative effect in A549. However, its potentiation nearly disappeared when the cells were transfected with siRNAs against PARP-1. Oral administration of AMF (100  mg/kg), combined with CBP, remarkably inhibited A549 tumor growth and ki67 expression, and increased apoptosis compared with CBP-alone group. CONCLUSION: All results suggest that AMF can be a potential PARP-1 inhibitor and a candidate adjuvant agent to boost the anticancer effect of CBP in NSCLC.

Rapid screening of non-steroidal anti-inflammatory drugs illegally added in anti-rheumatic herbal supplements and herbal remedies by portable ion mobility spectrometry.

In this work, for the first time, a high-performance ion mobility spectrometry with electrospray ionization (ESI-HPIMS) method has been employed as a rapid screening tool for the detection of acetaminophen, ibuprofen, naproxen, diclofenac sodium and indomethacin illegally added in anti-rheumatic herbal supplements and herbal remedies. Samples were dissolved and filtered through a 0.45µm microporous membrane, then the filtrate was directly injected into the high-performance ion mobility spectrometry for analysis. Using this approach, the screening of illegal additions can be accomplished in as rapid as two to three minutes with no pretreatment required. The proposed method provided a LOD of 0.06-0.33µgmL-1, as well as a good seperation of the five NSAIDs. The precision of the method was 0.1-0.4% (repeatability, n=6) and 0.9-3.3% (reproducibility, n=3). The proposed method appeared to be simple, rapid and highly specific, thus could be effective for the in-situ screening of NSAIDs in anti-rheumatic herbal supplements and herbal remedies.

Graphene oxide adsorbent based dispersive solid phase extraction coupled with multi-pretreatment clean-up for analysis of trace aflatoxins in traditional proprietary Chinese medicines.

Graphene oxide (GO)-based dispersive solid phase extraction (D-SPE) method combined with multi-step preparation has been proposed for the evaluation of trace aflatoxins in proprietary Chinese medicines (PCM). After being extracted by methanol, the sample was purified based on multi-step preparation, including dehydration with MgSO4/NaCl and cleanup with neutral alumina. Then GO was used as an adsorbent in D-SPE method for further preconcentration of aflatoxins prior to high performance liquid chromatography-fluorescence detection. The selected conditions were investigated. The Box-Behnken design (BBD) was used to optimize factors affecting adsorption procedure. Under the optimized conditions, good linear relationships had been achieved with the correlation coefficient (R2) varying from 0.9904 to 0.9990. The LODs and LOQs were ranging from 0.020 to 0.041ng/mL and 0.061 to 0.125ng/mL, respectively. The results of the recoveries were 74.0-102.7% for the four aflatoxins, while the precisions from 1.8% to 7.2% were obtained, which indicated that the method was suitable for the analysis of aflatoxins in PCM.

STAT3 as a potential therapeutic target in ALDH+ and CD44+/CD24+ stem cell-like pancreatic cancer cells.

Persistent activation of signal transducers and activators of transcription 3 (STAT3) is commonly detected in many types of cancer including pancreatic cancer. Whether STAT3 is activated in stem cell-like pancreatic cancer cells and the effect of STAT3 inhibition, is still unknown. Flow cytometry was used to isolate pancreatic cancer stem-like cells which are identified by both aldehyde dehydrogenase (ALDH)-positive (ALDH+) as well as cluster of differentiation (CD) 44-positive/CD24-positive subpopulations (CD44+/CD24+). STAT3 activation and the effects of STAT3 inhibition by STAT3 inhibitors, LLL12, FLLL32, and Stattic in ALDH+ and CD44+/CD24+ cells were examined. Our results showed that ALDH+ and CD44+/CD24+ pancreatic cancer stem-like cells expressed higher levels of phosphorylated STAT3, an active form of STAT3, compared to ALDH-negative (ALDH-) and CD44-negative/CD24-negative (CD44-/CD24-) pancreatic cancer cells, suggesting that STAT3 is activated in pancreatic cancer stem-like cells. Small molecular STAT3 inhibitors inhibited STAT3 phosphorylation, STAT3 downstream target gene expression, cell viability, and tumorsphere formation in ALDH+ and CD44+/CD24+ cells. Our results indicate that STAT3 is a novel therapeutic target in pancreatic cancer stem-like cells and inhibition of activated STAT3 in these cells by STAT3 inhibitors may offer an effective treatment for pancreatic cancer.

Nucleocytoplasmic trafficking of the Syk protein tyrosine kinase.

The protein tyrosine kinase Syk couples the B-cell receptor (BCR) for antigen to multiple intracellular signaling pathways and also modulates cellular responses to inducers of oxidative stress in a receptor-independent fashion. In B cells, Syk is found in both the nuclear and cytoplasmic compartments but contains no recognizable nuclear localization or export signals. Through the analysis of a series of deletion mutants, we identified the presence of an unconventional shuttling sequence near the junction of the catalytic domain and the linker B region that accounts for Syk's subcellular localization. This localization is altered following prolonged engagement of the BCR, which causes Syk to be excluded from the nucleus. Nuclear exclusion requires the receptor-mediated activation of protein kinase C and new protein synthesis. Both of these processes also potentiate the activation of caspase 3 in cells in response to oxidative stress in a manner that is dependent on the localization of Syk outside of the nucleus. In contrast, restriction of Syk to the nucleus greatly diminishes the stress-induced activation of caspase 3.