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Calcification of bones is the critical process of bone development in birds, which is very important for sustaining the normal biological function of bones. Light is one of the vital factors affecting bone development, but whether light intensity affects bone calcification and the underlying mechanism is still unknown. In this study, we used duck sternum as a model to analyze the calcification process under different light regimes. In addition, the underlying mechanism was also illustrated by integrating metabolomics and transcriptome methods. The experiment lasted from 14 to 51 d of duck age. The control group (LP1) kept light intensity 2 lx during the whole experiment. The two light supplement groups (LP2, LP3) were given light with the intensity of 70 lx at different time (14-29 d for LP2, 14-43 d for LP3). Samples were collected at 52 d of duck age. Sternal calcification analysis showed no significant difference in proportion of area of cartilage matrix and trabecular bone in keel tissue among the 3 groups, but the degree of keel calcification in LP3 was higher than in the other 2 groups. Serum metabolomics showed 32 and 28 differentially accumulated metabolites (DAMs) in the 2 comparison groups, LP1 vs. LP3 and LP1 vs. LP2, respectively. Carboxylic acids and derivatives were the most abundant among the DAMs. Sternal transcriptome analysis showed 231 differentially expressed genes (DEGs), including 177 upregulated genes and 54 downregulated genes in group LP1 vs. LP3, and 22 DEGs in group LP1 vs. LP2. Protein-protein interaction (PPI) network analysis on DEGs between LP1 and LP3 showed that genes BTRC, GLI1, BMP4, and FOS were in the core position of the interaction network, and are also involved in bone development. KEGG pathway analysis of DAMs and DEGs showed that differences in Hedgehog signaling pathway, MAPK signaling pathway, apoptosis, energy metabolism, and amino acid metabolism following light treatment seem likely to have contributed to the observed difference in calcification of duck sternum., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

To explored the difference of goose fatty liver formation induced-by different types of sugar from the intestinal physiology and the gut microflora, an integrated analysis of intestinal physiology and gut microbiota metagenomes was performed using samples collected from the geese including the normal-feeding geese and the overfed geese which were overfed with maize flour or overfeeding dietary supplementation with 10% sugar (glucose, fructose or sucrose, respectively), respectively. The results showed that the foie gras weight of the fructose group and the sucrose group was heavier (P < 0.05) than other groups. Compared with the control group, the ileum weight was significantly higher (P < 0.01), and the cecum weight was significantly lower in the sugar treatment groups (P < 0.001). Compared with the control group, the ratio of villi height to crypt depth in the fructose group was the highest in jejunum (P < 0.05); the trypsin activity of the ileum was higher in the fructose group and the sucrose group (P < 0.05). At the phylum level, Firmicutes, Proteobacteria and Bacteroidetes were the main intestinal flora of geese; and the abundance of Firmicutes in the jejunum was higher in the sugar treatment groups than that of the maize flour group. At the genus level, the abundance of Lactobacillus in the jejunum was higher (P < 0.05) in the sugar treatment groups than that of the maize flour group. In conclusion, forced-feeding diet supplementation with sugar induced stronger digestion and absorption capacity, increased the abundance of Firmicutes and Bacteroidetes and the abundance of Lactobacillus (especially fructose and sucrose) in the gut. So, the fructose and sucrose had higher induction on hepatic steatosis in goose fatty liver formation., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

To have a better understanding of how the "gut-liver axis" mediates the lipid deposition in the liver, a comparison of overfeeding influence on intestine physiology and microbiota between Gang Goose and Tianfu Meat Goose was performed in this study. After force-feeding, compared with Gang Goose, Tianfu Meat Goose had better fat storage capacity in liver (397.94 vs. 166.54 for foie gras weight (g), P < 0.05; 6.37 vs. 2.92% for the ratio of liver to body, P < 0.05; 60.01 vs. 46.64% for fat content, P < 0.05) and the less subcutaneous adipose tissue weight (1240.96 g vs. 1440.46 g, P < 0.05). After force-feeding, the digestion-absorption capacity of Tianfu Meat Goose was higher than that of Gang Goose (5.56 vs. 3.64 and 4.63 vs. 3.68 for the ratio of villus height to crypt depth in duodenum and ileum, respectively, P < 0.05; 1394.96 vs. 782.59 and 1314.76 vs. 766.17 for the invertase activity (U/mg-prot), in duodenum and ileum, respectively, P < 0.05; 6038.36 vs. 3088.29 and 4645.29 vs. 3927.61 for the activity of maltase (U/mg-prot), in duodenum and ileum, respectively, P < 0.05). Force-feeding decreased the gene expression of Escherichia coli in the ileum of Tianfu Meat Goose; force-feeding increased the number of gut microbiota Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction band in Tianfu Meat Goose and decreased the number in Gang Goose. In conclusion, compared with Gang Goose, the lipid deposition in the liver and the intestine digestion-absorption capacity and stability were higher in Tianfu Meat Goose. Thereby, Tianfu Meat Goose is the better breed for foie gras production for prolonged force-feeding; Gang Goose possesses better fat storage capacity in subcutaneous adipose tissue. However, Gang Goose has lower gut stability responding to force-feeding, so Gang Goose is suited to force-feeding in a short time to gain the body weight and subcutaneous fat as an overfed duck for roast duck., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) produced by hepatocellular carcinoma (HCC) cells are responsible for the growth of HCC cells. Accumulating evidence shows that insulin-like growth factor-binding protein-3 (IGFBP-3) suppresses HCC cell proliferation in both IGF-dependent and independent manners. It's unknown, however, whether treatment with exogenous IGFBP-3 inhibits bFGF and PDGF production in HCC cells. The present study demonstrates that IGFBP-3 suppressed IGF-1-induced bFGF and PDGF expression while it does not affect their expression in the absence of IGF-1. To delineate the underlying mechanism, western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1 (EGR1) is involved in IGFBP-3 regulation of bFGF and PDGF. IGFBP-3 inhibition of type 1 insulin-like growth factor receptor (IGF1R), ERK and AKT activation is IGF-1-dependent. Furthermore, transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1, bFGF and PDGF expression. In conclusion, these findings suggest that IGFBP-3 suppresses transcription of EGR1 and its target genes bFGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation. It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation, suggesting that IGFBP-3 could be a target for the treatment of HCC., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Unlabelled: Phosphatidylinositol-3 kinases (PI3K)-Protein kinase B (Akt)-mammalian target of rapamycin (mTOR) pathway plays an important role in the synthesis and secretion of triacylglycerol. However, the mechanism of PI3K-Akt-mTOR pathway in regulating lipid metabolism of goose liver was poorly understood. The purpose of this study was to determine how PI3K-Akt-mTOR pathway regulating lipid metabolic homeostasis in goose hepatocytes. Goose primary hepatocytes were treated with different PI3K-Akt-mTOR signal inhibitors (LY294002, rapamycin and NVP-BEZ235) for 24 h. The results showed that these inhibitors evidently inhibited PI3K-Akt-mTOR downstream signaling. Meanwhile, these PI3K-Akt-mTOR inhibitors reduced intracellular lipid accumulation, decreased the mRNA expression and protein content of genes involved in the de novo fatty acid synthesis, while increased the transcriptional and protein level of key factors involved in fatty acid oxidation and very low density lipoprotein (VLDL) assembly and secretion., Conclusion: These findings suggested that the reduction of lipids accumulation induced-by inhibiting PI3K-Akt-mTOR pathway was closely linked to the decrease of lipogenesis, the increase of fatty acids oxidation, and the increase of VLDL assembly and secretion in goose hepatocytes.

Cryopreservation technology regarding banded coral shrimp (Stenopus hispidus) embryos is important as it could improve cultivation and preservation of the species. The development of this technology is to reduce collections of this species from the wild, thus preventing damage to coral reefs. This study investigated the tolerance of different developmental stages of S. hispidus embryos in response to low temperature in the presence or absence of cryoprotectant. Embryos undergoing three stages of embryonic development (eye-formation, heart beat and pre-hatch stage) were selected and exposed to 5, 0 and -5°C in cryoprotectant solutions of 1 or 2M methanol for 1, 2, 4, 6, 8, 16 and 32h. Embryo survival was evaluated based on their hatching percentage. In experiments on the effect of different concentrations of methanol on chilling sensitivity of embryos, it was determined that methanol at 1M methanol reduced the chilling sensitivities of embryos most effectively when compared to the other tested concentrations. Experiments regarding the chilling sensitivity of embryos in different developmental stages indicated that pre-hatch stage embryos were more resistant to subzero temperatures than early stage embryos; they tolerated the 32h exposure at 5 and 0°C without a loss in survival. The study also indicated that late stage embryos are considered to be resistant to chilling, and that pre-late stage embryos are better candidate for cryopreservation., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Myostatin (MSTN) plays a key role in the negative regulation of muscle growth and development during embryogenesis. The MSTN genes have different genetic characteristics in vertebrates: sole gene in mammals, gene duplication in fish, and alternative splicing in birds. To investigate the alternative splicing sites and developmental expression patterns of the duck MSTN genes, the mRNA and genome sequences were cloned, and the expression patterns were detected during breast muscle and leg muscle development by real-time PCR. In our study, four alternatively spliced forms of MSTN mRNA were found in the developing skeletal muscle of Peking duck, including two novel alternatively spliced transcripts, MSTN-c and MSTN-d. As a result of alternative splicing at the common GT-AG processing sites, MSTN-b and MSTN-c retained only the N-terminal TGFβ-propeptide superfamily domains. However, MSTN-d was not missing these domains, in contrast to MSTN-a. The real-time PCR results showed that there was no significant difference between breast muscle and leg muscle in MSTN-a mRNA expression, also in MSTN-b and MSTN-c. MSTN-a and MSTN-b have significant higher expressions than MSTN-c and MSTN-d, suggesting that they play the major role during embryo muscle development., (Copyright © 2011 Elsevier Inc. All rights reserved.)

In this study, we investigated the role of liver X receptor (LXR) activation in hepatic assembly and in the secretion of very low density lipoprotein-triglycerides in goose primary hepatocytes. Goose primary hepatocytes were isolated and treated with the LXR agonist T0901317. Total triglyceride accumulation, intracellular and extracellular triglyceride concentrations, extracellular very low density lipoprotein concentration, and gene expression levels of LXRα, microsomal triglyceride transfer protein, acyl coenzyme A:diacylglycerol acyltransferase (DGAT) 1, and DGAT2 were measured in primary hepatocytes. We found a dose-dependent upregulation of total and intracellular TG accumulation when using 0, 0.01, 0.1, 1, and 10 μM T0901317, but the extracellular triglyceride and very low density lipoprotein concentrations were dose dependent only when the T0901317 concentration was below 1 μM; as compared with 1 μM T0901317, 10 μM T0901317 had an inhibiting effect (P < 0.05). The mRNA levels of all the detected genes increased in the presence of T0901317. The change in LXRα and DGAT1 was dose dependent, and the mRNA levels of microsomal triglyceride transfer protein and DGAT2 increased with a T0901317 concentration up to 1 μM, but decreased when treated with 10 μM T0901317 (P < 0.05). In conclusion, the secretion of very low density lipoprotein plays a role in pharmacologically activating the LXR-induced development of hepatocellular steatosis in geese.

Electrospray ionization (ESI) has been an indispensable ion generation technique for mass spectrometric analysis of biopolymers such as intact proteins and protein digests operated at atmospheric pressure. Since its advent in 1998, atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI) quickly became a popular alternative for the analysis of peptides. Although AP-MALDI sources typically share the same vacuum interface and ion transmission hardware with ESI, it is generally found that ESI is superior in detection sensitivity. Here we present a method based on solid phase extraction and elution with surface-functionalized diamond nanocrystals (which we previously referred to as "SPEED") that not only streamlines AP-MALDI mass spectrometric analyses of peptides and other small biomolecules under typical operational conditions but also outruns ESI in ultimate detectable concentration by at least one order of magnitude.

Background: Endobronchial neodymium:yttrium aluminum garnet (Nd:YAG) laser therapy has been shown to be safe and effective in palliating large airway malignancies. The role of multimodal therapy with stenting, brachytherapy, chemotherapy, and radiotherapy together with Nd:YAG laser therapy is unclear., Aim: This study aims to confirm the safety and effectiveness of Nd:YAG laser therapy in the contemporary setting and to investigate the effectiveness of multimodal therapy compared with laser alone., Methods: One hundred fifty-three Nd:YAG laser treatments on 110 patients between 1999 and 2004 were reviewed retrospectively. Symptom scores for dyspnoea, hemoptysis, and cough before and after the procedure were compared. Survival and time to reintervention were analyzed using the Kaplan-Meier method. Multimodality treatment was compared with Nd:YAG laser therapy alone to determine differences in survival and time to reintervention. p values less than 0.05 were considered significant., Results: There were no operative mortalities directly caused by laser intervention, although 30-day mortality was 6.5%. Deaths were attributable to progression of advanced neoplastic processes rather than laser intervention, and 6.5% of patients had some postoperative morbidity. After Nd:YAG laser intervention, 76% of patients reported improvement to dyspnoea, 94% for hemoptysis, and 75% for cough. Median survival after Nd:YAG laser treatment was 6.64 months; 21% of patients required repeated laser treatment. Compared with Nd:YAG laser treatment alone, multimodality treatments significantly prolonged median time to reintervention by 1.7 months (p = 0.002) and prolonged median survival by 4.9 months (p < 0.001) in patients with NSCLC., Conclusion: Nd:YAG laser intervention is safe and effective for palliation of endobronchial malignancies. In most cases, it only needs to be performed once. Compared with Nd:YAG laser therapy alone, multimodal treatment prolonged survival.

In order to understand the changes in protein dynamics that occur in the final stages of protein folding, we have used neutron scattering to probe the differences between a protein in its folded state and the molten globule states. The internal dynamics of bovine alpha-lactalbumin (BLA) and its molten globules (MBLA) have been examined using incoherent, quasielastic neutron scattering (IQNS). The IQNS results show length scale dependent, pico-second dynamics changes on length scales from 3.3 to 60 A studied. On shorter-length scales, the non-exchangeable protons undergo jump motions over potential barriers, as those involved in side-chain rotamer changes. The mean potential barrier to local jump motions is higher in BLA than in MBLA, as might be expected. On longer length scales, the protons undergo spatially restricted diffusive motions with the diffusive motions being more restricted in BLA than in MBLA. Both BLA and MBLA have similar mean square amplitudes of high frequency motions comparable to the chemical bond vibrational motions. Bond vibrational motions thus do not change significantly upon folding. Interestingly, the quasielastic scattering intensities show pronounced maxima for both BLA and MBLA, suggesting that "clusters" of atoms are moving collectively within the proteins on picosecond time scales. The correlation length, or "the cluster size", of such atom clusters moving collectively is dramatically reduced in the molten globules with the correlation length being 6.9 A in MBLA shorter than that of 18 A in BLA. Such collective motions may be important for the stability of the folded state, and may influence the protein folding pathways from the molten globules., (Copyright 2000 Academic Press.)