Osteocalcin activates lipophagy via the ADPN-AMPK/PPARα-mTOR signaling pathway in chicken embryonic hepatocyte.

Fatty liver hemorrhage syndrome (FLHS) is the leading cause of noninfectious mortality in caged layers worldwide. Osteocalcin (OCN) is a protein secreted by osteoblasts, and its undercarboxylated form (ucOCN) acts as a multifunctional hormone that protects laying hens from FLHS. Lipophagy is a form of selective autophagy that breaks down lipid droplets (LDs) through lysosomes, and defective lipophagy is associated with FLHS. The aim of this study was to investigate the effects of ucOCN on the lipophagy of chicken embryonic hepatocytes and associated the function of the adiponectin (ADPN) signaling pathway. In this study, chicken embryonic hepatocytes were divided into 5 groups: control (CONT), fat emulsion (FE, 10% FE, v/v), FE with ucOCN at 1 ng/mL (FE-LOCN), 3 ng/mL (FE-MOCN), and 9 ng/mL (FE-HOCN). In addition, 4 µM AdipoRon, an adiponectin receptor agonist, was used to investigate the function of ADPN. The results showed that compared with CONT group, FE promoted the levels of phosphorylation of mammalian target of rapamycin (p-mTOR) (P < 0.05) and decreased the mRNA expression of ADNP receptors (AdipoR1 and AdipoR2). Compared with FE group, 3 and 9 ng/mL ucOCN inhibited the levels of autophagy adaptor p62 and p-mTOR (P < 0.05), increased the ratios of LC3-II/LC3-I (P < 0.05) and phosphorylated adenosine 5'-monophosphate-activated protein kinase (p-AMPK)/AMPK (P < 0.05), as well as the levels of peroxisome proliferator-activated receptor α (PPAR-α) and ADPN (P < 0.05). In addition, ucOCN at the tested concentrations increased the colocalization of LC3 and LDs in fatty hepatocytes. Administrated 4 µM AdipoRon activated AdipoR1 and AidpoR2 mRNA expression (P < 0.05), decreased the concentrations of triglyceride (P < 0.05), without effects on cell viability (P > 0.05). AdipoRon also increased the LC3-II/LC3-I ratio (P < 0.05) and the levels of p-AMPK/AMPK and PPAR-α (P < 0.05). In conclusion, the results reveal that ucOCN regulates lipid metabolism by activating lipophagy via the ADPN-AMPK/PPARα-mTOR signaling pathway in chicken embryonic hepatocytes. The results may provide new insights for controlling FLHS in laying hens.

Osteocalcin reduces fat accumulation and inflammatory reaction by inhibiting ROS-JNK signal pathway in chicken embryonic hepatocytes.

Osteocalcin (OCN) has a function in preventing fatty liver hemorrhagic syndrome (FLHS) in poultry. The aim of this study was to investigate the effects of OCN on fat emulsion stimulated chicken embryonic hepatocytes and related signaling pathways. The primary chicken embryonic hepatocytes were isolated from the incubated 15-day (E15) pathogen free eggs and cultured with dulbecco's modified eagle medium (DMEM). After the hepatocyte density reached 80%, the cells were divided into 5 groups: control group (CONT), fat emulsion group (FE, 10% FE, v/v), FE with ucOCN at 1 ng/mL (FE-LOCN), 3 ng/mL (FE-MOCN), and 9 ng/mL (FE-HOCN). In addition, 2 mM N-Acetyl-L-cysteine (NAC) a reactive oxygen species (ROS) scavenger, and 5 µM SP600125, a Jun N-terminal kinase (JNK) inhibitor, were added separately in to the DMEM with 10% FE to test effects of FE on the function of ROS-JNK signal pathway. The number of hepatocytes, cell ultra-microstructure, viability, and apoptosis were detected after 48 h treatment, and the protein expressions and enzyme concentrations were detected after 72 h treatment. The results showed that, compared to the control group, FE increased the triglyceride (TG) concentration and lipid droplets (LDs) in chicken embryonic hepatocytes (P < 0.05), and induced hepatocytic edema with obviously mitochondrial swelling, membrane damage, and cristae rupture. FE also decreased ATP concentration, increased ROS concentrations and mitochondrial DNA (mtDNA) copy number, promoted inflammatory interleukin-1 (IL-1), IL-6, tumor necrosis factor-alpha (TNF-α) concentrations and hepatocytic apoptosis rate, and raised phospho-c-Jun N-terminal kinase (p-JNK) protein expressions. Compared to the FE group, ucOCN significantly increased hepatocyte viability, reduced hepatocytic TG concentrations and LDs numbers, and alleviated hepatocytic edema and mitochondrial swelling. Furthermore, ucOCN significantly decreased ROS concentrations, increased ATP concentrations, reduced IL-1, IL-6, TNF-α concentrations and hepatocytic apoptosis rate, and inhibited p-JNK protein expressions (P < 0.05). NAC had the similar functions of ucOCN reduced the ROS concentration and inhibited the TNF-α protein expression and p-JNK/JNK ration. Similarly, SP600125 reduced p-JNK/JNK protein expression, IL-1, IL-6, TNF-α, and TG concentrations without effects on ROS concentration and hepatocytic apoptosis. These results suggest that ucOCN alleviates FE-induced mitochondrial damage, cellular edema, and apoptosis of hepatocytes. These results reveal that the functions of ucOCN in reducing fat accumulation and inflammatory reaction in chicken embryonic hepatocytes are mostly via inhibiting the ROS-JNK signal pathway.

Bacillus subtilis inhibits intestinal inflammation and oxidative stress by regulating gut flora and related metabolites in laying hens.

Bacillus subtilis is one of the most popular commercial probiotics used in farm animal production. However, its potential mechanisms are not very clear. The aim of this study was to investigate the effects of dietary Bacillus subtilis on intestinal histomorphology, innate immunity, microbiota composition, transcriptomics, and related metabolomics. Twenty-four 48-week-old Lohman Pink-shell laying hens were randomly divided into two groups: a basic diet and the basic diet supplemented with Bacillus subtilis (0.5 g/kg) for a 9-week experiment. At the end of the experiment, tissues of the duodenum, ileum, and jejunum as well as cecal content of each bird were collected for microstructure, PCR, transcriptome, metabolome, and 16S rRNA analyses. The results showed that dietary Bacillus subtilis supplement had no effect on the intestinal microstructure. However, Bacillus subtilis increased mRNA expression of tight junction protein occludin (P < 0.05), while reduced mRNA expression of lipopolysaccharide-induced TNF factor (P < 0.01) in the duodenum. Moreover, transcriptomic results indicated that most of Bacillus subtilis supplement-induced differential genes were associated with inflammation and immunity, including cytochrome b-245 beta chain, transferrin, and purinergic receptor P2X 7, resulting in a decrease in Malondialdehyde level (P < 0.05) in the duodenum. In addition, at the genus level, Bacillus subtilis supplement enriched the potential beneficial bacteria, Candidatus_Soleaferrea (P = 0.02) but inhibited the harmful bacteria including Lachnospiraceae_FCS020_group, Ruminiclostridium, Lachnospiraceae_UCG-010, and Oxalobacter. Metabolomic results revealed that N-Acetylneuraminic acid and ADP were increased by fed Bacillus subtilis. These results suggest that dietary Bacillus subtilis could inhibit gut inflammation and improve antioxidative status and barrier integrity of the duodenum via regulating gut microbial composition in laying hens.

[Electroencephalogram power development of cognitive function at age 7 to 12 years: a comparative study between attention deficit hyperactivity disorder and healthy children].

Objective: To compare brain electrical cognitive tasks and brain development between study about 7 to 12 years old attention deficit hyperactivity disorder (ADHD) and normal children. Method: Prospectic case-control study was used. A total of 110 children with ADHD (63 boys and 47 girls) and 116 normal children (66 boys and 50 girls), were enrolled in this study. The electroencephalogram (EEG) was recorded when attention tasks were conducted, the EEG power was extracted from the original data and comparatively analyzed the absolute power (θ, α, ß spectrum) and relative power (θ/total, α/total, θ/α, θ/ß). Result: (1) Absolute power: ADHD children θ absolute power was higher than that of normal children in Pz lead ((52±28)vs. (40±30)µV2, t=3.906, P<0.05), with statistical significance. (2) Relative power: θ/total, θ/α, θ/ß in ADHD are higher than normal children(0.23±0.07 vs. 0.20±0.05, 1.35±0.76 vs. 1.00±0.56, 4.75±2.49 vs. 3.56±2.08, t=2.900 and 3.954 and 3.901, P=0.004 and 0.000 and 0.000), α/total in ADHD is lower (0.21±0.09 vs. 0.24±0.10, t=-2.517, P=0.013). (3) The comparative study of the development of EEG power θ/ß between ADHD and normal children showed age-related correlation in both groups (r=-0.378 and -0.398, P=0.000 for both). Conclusion: ADHD children's EEG power on slow spectrum was higher than that of the normal children, it was more significant in the parietal region than in frontal region. With the increase of age, the θ relative power in ADHD and normal children gradually declined, in the normal children it linearly related, but in ADHD there was no significant regularity. θ/ß can be used as a sensitive index to assess ADHD children's cognitive function.

[Psychopathological characteristics in patients with deviation of nasal septum: a preliminary analysis].

Objective: To investigate the psychopathological characteristics in patients with deviation of nasal septum. Methods: Between May 2015 and December 2015, fourty-four patients with deviated nasal septum and 37 patients with vocal cord polyp as control were included in this study. Psychological characteristics were evaluated by a series of questionnaire instruments including symptom checklist-90 (SCL-90), self-rating depression scale (SDS) and self-rating anxiety scale (SAS). Visual analogue scale (VAS) and rhinomanometry through front nostril were used to evaluate nasal symptom. The correlation between psychological characteristics and nasal symptom was evaluated. SPSS 20.0 software was used to analyze the data. Results: The SCL-90 score in nasal septal deviation group was 130.4±48.3. The total score and total average score of SCL-90 had no significant difference between nasal septal deviation group and the Chinese standard or control group(t value was 0.469, 0.112, 1.575, 1.564, respectively, all P>0.05). The scores of somatization, depression and anxiety factors in nasal septal deviation group were higher than control group (t value was 2.380, 2.133, 1.969, respectively, all P<0.05). The proportion of positive patients in these three factors between nasal septal deviation group and control group had significant differences (χ2 value was 11.585, 9.610, 5.429, respectively, all P<0.05). The scores of SDS and SAS in nasal septal deviation group were 46.0±10.6 and 43.0±10.2, which were higher than that in the Chinese standard and control group (t value was 5.342, 6.236, 1.476, 3.013, respectively, all P<0.05). There were 9 patients companying with depression or anxiety (20.5%, 20.5%, respectively) and 5 patients companying with depression and anxiety in nasal septal deviation group (11.4%). There were positive correlation not only between the scores of SDS and the depression factor of SCL-90 but also between the scores of SAS and the anxiety factor of SCL-90 (Z=0.415, P=0.005, Z=0.445, P=0.002, respectively). The scores of SDS and SAS had positive correlation (Z=0.392, P=0.008). The VAS score of nasal obstruction was 6.0±3.2. The rhinomanometry in inspiratory and expiratory phase were (0.202±0.140) kPa·S/cm3 and (0.230±0.161) kPa·S/cm3. Besides the positive correlation between the rhinomanometry in inspiratory phase and SDS (Z=0.332, P=0.045), the psychological scores, including SCL-90 score, depression, anxiety factors score, SAS and SDS, had no correlation with VAS scores and rhinomanometry (r value was -0.030, -0.052, -0.026, 0.107, 0.185, 0.066, 0.160, 0.203, respectively, all P>0.05). Conclusions: High prevalence of depression and anxiety is found in patients with deviation of nasal septum. The SCL-90 score is consistent with SDS and SAS. Besides the positive correlation between the rhinomanometry in inspiratory phase and SDS, the psychological scores (SCL-90 score, depression, anxiety factors score, SAS and SDS) have no correlation with VAS score and rhinomanometry.

Association between serum 25-hydroxyvitamin D levels measured 24 hours after delivery and postpartum depression.

OBJECTIVE: To assess the possible relationship between serum levels of 25[OH]D (25-hydroxyvitamin D) collected 24 hours after delivery and postpartum depression in a Chinese cohort sample. DESIGN: Cohort study. SETTING: One city hospital in Beijing, China. POPULATION: Women delivering a full-term, singleton, live-born infant at one city hospital in Beijing between August 2013 and November 2013. METHODS: Women were enrolled immediately postpartum. A blood sample was obtained 24-48 hours after childbirth to test serum levels of 25[OH]D. Participation consisted of a visit to an obstetric unit 3 months after delivery. MAIN OUTCOME MEASURE: At 3 months' postpartum, women were screened for depression using the Edinburgh Postnatal Depression Scale (EPDS). The primary outcome measure was a prespecified EPDS score of ≥12. RESULTS: During the study period, 323 women were admitted. In all, 248 agreed to enrol and 213 completed 3 months' follow-up (21 were lost to follow-up and 14 withdrew). Of the 213 women who were included, 26 (12.2%) were considered to meet criteria for postpartum depression. Serum 25[OH]D levels in women with no postpartum depression were significantly higher than those in women with postpartum depression (P < 0.0001). Based on the receiver operating characteristic curve, the optimal cutoff value for serum 25[OH]D level as an indicator for screening for postpartum depression was estimated to be 10.2 ng/ml, with an area under the curve of 0.801 (95%CI 0.704-0.896). In multivariate analysis, there was an increased risk of postpartum depression associated with 25[OH]D levels ≤10.2 ng/ml (OR 7.17, 95%CI 3.81-12.94; P < 0.0001) after adjusting for possible confounders. CONCLUSION: Our study demonstrated that lower serum 25[OH]D levels were associated with postpartum depression. This association was independent of other possible variables.

Prognostic value of plasma neuroendocrine biomarkers in patients with acute ischaemic stroke.

Inflammation and activation of the neuroendocrine systems comprise important aspects of stroke pathophysiology. The present study investigated whether baseline plasma brain natriuretic peptide (BNP), N-terminal pro-brain natriuretic peptide (NT-proBNP), cortisol and copeptin levels on admission can predict short-term outcomes and mortality after acute ischaemic stroke. The study group consisted of 189 patients who had their first acute ischaemic stroke. Plasma levels of BNP, NT-proBNP, cortisol and copeptin were evaluated to determine their value with respect to predicting functional outcome and mortality within 3 months. As a result of cardiovascular and neurological investigations (including imaging techniques), lesion size, stroke subtype classification and clinical outcome after 3 months were determined. Plasma levels of BNP, NT-proBNP, cortisol and copeptin were associated with stroke severity, as well as short-term functional outcomes. After adjusting for all other significant outcome predictors, NT-proBNP, cortisol and copeptin remained as independent outcome predictors. In the receiver operating characteristic curve analysis, the biomarker panel (including BNP, NT-proBNP, cortisol and copeptin) predicted functional outcome and death within 90 days significantly more efficiently than the National Institute of Health Stroke Scale (NIHSS) or the biomarker alone. Copeptin showed a significantly greater discriminatory ability as a single biomarker compared to BNP, NT-proBNP, cortisol and NIHSS score. These results suggest that a biomarker panel may add valuable and time-sensitive prognostic information in the early evaluation of acute ischaemic stroke. This may provide a channel for interventional therapy in acute stroke.

Molecular characterization of foot-and-mouth disease virus isolated from ruminants in Taiwan in 1999-2000.

In 1999, 10 sporadic outbreaks of cattle foot-and-mouth disease (FMD) occurred in Taiwan. By the time, infection was limited to the Chinese yellow cattle (a native species of beef cattle in Mainland China), which did not develop vesicular lesions under field conditions. Five viruses isolates obtained from individual farms were confirmed to be the serotype O FMD virus (O/Taiwan/1999). During January-February 2000, however, this virus has spread to dairy cattle and goat herds, causing severe mortality in goat kids and vesicular lesions in dairy cattle. Partial nucleotide sequence of the capsid coding gene 1D (VP1) was determined for the virus isolates obtained in this study. Phylogenetic analysis of the VP1 sequences indicated that the O/Taiwan/1999 viruses shared 95-97% similarities to the virus strains isolated from the Middle East and India. The species susceptibility of the O/Taiwan/1999 virus was experimentally studied in several species of susceptible animals, showing that the virus did cause generalized lesions in dairy cattle and pigs, however, it would not cause vesicular lesions on the Chinese yellow cattle and the adult goats. These studies suggested that the O/Taiwan/1999 virus was a novel FMD virus of Taiwan and it presented various levels of susceptibility in cattle species.

Neoplastic response in Japanese medaka and channel catfish exposed to N-methyl-N'-nitro-N-nitrosoguanidine.

Japanese medaka (Oryzias latipes) and channel catfish (Ictalurus punctatus) were investigated for carcinogenic response following a 28-day, 3 x/wk pulse exposure to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Five-wk-old medaka were exposed at concentrations of 0, 0.5, and 1.0 mg/L, and 5-mo-old catfish at concentrations of 0, 0.1, and 0.5 mg/L. In medaka, a total of 19 tumors including 2 branchioblastomas, 6 thyroid follicular adenomas and 1 adenocarcinoma, and 11 subcutaneous fibrosarcomas were observed in 16 of 96 MNNG-exposed fish. In catfish, a total of 37 tumors including 4 squamous cell carcinomas and 16 papillomas, 3 lipomas, 1 fibroma, 1 osteosarcoma, 4 branchioblastomas, 6 thymic epithelial tumors, and 2 generalized lymphosarcomas were observed in 34 of 172 MNNG-exposed fish. The induction of neoplasms in medaka was primarily in the gill, thyroid, and subcutis of the cervical and trunk regions, whereas in catfish skin, thymus, oro-pharynx, and hemopoietic tissues were also commonly affected. In both species, the neoplastic response was considered to be related to direct exposure of the tissues to MNNG. Some of these tumors have not been reported in the literature in either natural or experimental fish. The results also suggest species-specific differences in carcinogenic response following MNNG exposure.