Pharmacokinetics of florfenicol in blunt-snout bream (Megalobrama amblycephala) at two water temperatures with single-dose oral administration.

The pharmacokinetics and residue elimination of florfenicol (FFC) and its metabolite florfenicol amine (FFA) were studied in healthy blunt-snout bream (Megalobrama amblycephala, 50 ± 10 g). The study was conducted with a single-dose (25 mg/kg) oral administration at a water temperature of 18 or 28°C, while in the residue elimination study, fish were administered at 25 mg/kg daily for three consecutive days by oral gavage to determine the withdrawal period (WDT) at 28°C. The FFC and FFA levels in plasma and tissues (liver, kidneys and muscle) were analysed using high-performance liquid chromatography (HPLC). A no-compartment model was used to analyse the concentration versus time data of M. amblycephala. In the two groups at 18 and 28°C, the maximum plasma concentration (Cmax ) of FFC was 5.89 and 6.21 µg/ml, while the time to reach Cmax (Tmax ) was 5.97 and 2.84 hr, respectively. These suggested that higher temperature absorbed more drug and more quickly at M. amblycephala. And the elimination half-life (T1/2 kß ) of FFC was calculated as 26.75 and 16.14 hr, while the total body clearance (CL) was 0.09 and 0.15 L kg-1  hr-1 , and the areas under the concentration-time curves (AUCs) were 265.87 and 163.31 µg hr/ml, respectively. The difference demonstrated that the elimination rate of FFC in M. amblycephala at 28°C was more quickly than that at 18°C. The results of FFA showed the same trend in tissues of M. amblycephala. After multiple oral doses (25 mg/kg daily for 3 days), the k (eliminate rate constant) of FFA in M. amblycephala muscle was 0.017, the C0 (initial concentration) was 3.07 mg/kg, and the WDT was 10 days (water temperature 28°C).

Regulatory Effect of Bacillus subtilis on Cytokines of Dendritic Cells in Grass Carp (Ctenopharyngodon Idella).

Bacillus subtilis is a common group of probiotics that have been widely used in the feed industry as they can increase host resistance to pathogens and balance the immune response. However, the regulatory mechanism of Bacillus subtilis on the host immune system remains unclear in teleosts. In this study, we isolated and enriched dendritic cells from white blood cells (WBCs), and then stimulated them with Bacillus subtilis. Morphological features, specific biological functions, and authorized functional molecular markers were used in the identification of dendritic cells. Subsequently, we collected stimulated cells at 0, 4, and 18 h, and then constructed and sequenced the transcriptomic libraries. A transcriptome analysis showed that 2557 genes were up-regulated and 1708 were down-regulated at 4 h compared with the control group (|Fold Change| ≥ 4), and 1131 genes were up-regulated and 1769 were down-regulated between the cells collected at 18 h and 4 h (|Fold Change| ≥ 4). Gene Ontology (GO) annotations suggested many differentially expressed genes (DEGs) (p < 0.05 and |Fold Change| ≥ 4) were involved in immune-related biological functions including immune system progress, cytokine receptor binding, and cytokine binding. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the cytokine⁻cytokine receptor interaction pathways were significantly enriched at both time points (p < 0.05), which may play a key role in the response to stimulation. Furthermore, mRNA expression level examination of several pro-inflammatory cytokines and anti-inflammatory cytokines genes by quantitative real-time polymerase chain reaction (qRT-PCR) indicated that their expressions can be significantly increased in Bacillus subtili, which suggest that Bacillus subtilis can balance immune response and tolerance. This study provides dendritic cell (DC)-specific transcriptome data in grass carp by Bacillus subtilis stimulation, allowing us to illustrate the molecular mechanism of the DC-mediated immune response triggered by probiotics in grass carp.

Glucagon-like peptide-1 attenuates liver injury in the rat diabetes model.

The aim of present study is to examine the effect of glucagon-like peptide-1(GLP-1) on diabetes-induced liver injury and explore detailed mechanisms of GLP-1 hepatoprotective effect. 150 male Sprague-Dawley rats were randomly assigned into three groups with equal number, including Sham group, diabetes group and GLP-1 intervention group. Diabetes rat model was performed with intraperitoneal injection of streptozotocin (STZ, 65mg/kg). Fasting blood-glucose of rat model was assessed at 72h after STZ injection to verify diabetes rat model. Rats in Sham group were normally fed. Rats in GLP-1 intervention group received 2 ng/kg GLP-1 intervention, at 2, 4, 6 and 8 weeks after intervention, TUNEL staining were performed to examine apoptosis of liver tissue. PCR and Western blot were performed to examine insulin, GLP-1R, autophagy-associated gene and HDAC-1. Compared with diabetes group, insulin expression of GLP-1 intervention group increased significantly (P<0.05). TUNEL staining at different time showed apoptosis levels of liver tissues were reduced gradually after GLP-1 intervention (P<0.05). Compared with diabetes groups, the expressions of BCL2 and GLP-1R were increased, while the levels of caspase3 and LC3 were reduced in GLP-1 intervention group (P<0.05). GLP-1 treatment decreased levels of phosphorylated AKT, phosphorylated ERK1/2, and HDAC6 in liver tissues (P<0.05). GLP-1 treatment alleviated diabetes-induced liver injury via regulating autophagy. The mechanism of GLP-1 hepatoprotective effect could be via GLP-1R-ERK1/2-HDAC6 signaling pathway.

Oxidative stress and free radical damage in patients with acute dipterex poisoning.

OBJECTIVE: To investigate whether acute dipterex poisoning (ADP) may cause oxidative stress and free radical damage in the bodies of acute dipterex poisoning patients (ADPPs), and to explore the mechanisms by which ADP may cause oxidative stress and free radical damage. METHODS: Fifty ADPPs and fifty healthy adult volunteers (HAVs) whose ages, gender and others were matched with the ADPPs were enrolled in a randomized controlled study, in which concentrations of nitric oxide (NO), vitamin C (VC), vitamin E (VE) and beta-carotene (beta-CAR) in plasma as well as concentration of lipoperoxide (LPO), and activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and acetylcholinesterase (AChE) in erythrocytes were determined by spectrophotometric analytical methods. RESULTS: Compared with the average values of experimental parameters in the HAVs group, the average values of plasma NO and erythrocyte LPO in the ADPPs group were significantly increased (P<0.0001), while those of plasma VC, VE and beta-CAR as well as erythrocyte SOD, CAT, GPX and AChE in the ADPPs group were significantly decreased (P<0.0001). Bivariate correlation analysis and partial correlation analysis suggested that when NO and LPO values were increased, and VC, VE, beta-CAR, SOD, CAT and GPX values were decreased in the ADPPs, AChE value was decreased gradually in the ADPPs (P<0.001-0.0001). Reliability analysis of experimental parameters reflecting oxidative stress and free radical damage in the ADPPs showed that the reliability coefficient (8 items) alpha=0.6909, and the standardized item alpha=0.8574. CONCLUSION: The findings in the present study suggest that ADP can cause oxidative stress and free radical damage, and inhibit markedly erythrocyte acetylcholinesterase activity in ADPPs.

Influence of hypertension, lipometabolism disorders, obesity and other lifestyles on spontaneous intracerebral hemorrhage.

OBJECTIVE: To investigate whether hypertension, abnormal lipometabolism, obesity, cigarette smoking and alcohol drinking affect the intracerebral hemorrhagic volumes (IHV) in patients with spontaneous intracerebral hemorrhage (SIHP), and to explore the roles of these factors in spontaneous intracerebral hemorrhage (SIH). METHODS: Five hundred patients with acute SIH and 200 healthy adult volunteers (HAV) were enrolled in a study of independently randomized controlled design, in which the levels of systolic pressure (SP) and diastolic pressure (DP), and total cholesterol (TCH), triacylglycerols (triglycerides, TG), high density lipoprotein cholesterol (HDL-CH), low density lipoprotein cholesterol (LDL-CH) in serum as well as the level of erythrocytic membrane cholesterol (EM-CH) were measured, and the body mass index (BMI), daily cigarette smoking consumption (DCSC) and daily pure alcohol consumption (DPAC) were calculated. RESULTS: Compared with the average parameters in the HAV group, those of SP, DP, TG, LDL-CH and BMI in the SIHP group were significantly increased (P < 0.0001), while those of HDL-CH and EM-CH were significantly decreased (P < 0.0001). The linear regression and correlation analysis showed that with increased SP, DP, LDL-CH, BMI, DCSC, DPAC and aging as well as decreased HDL-CH and EM-CH, the IHV levels in SIHP were increased gradually (P < 0.0001-0.01). The linear stepwise regression analysis suggested that there existed a close correlation among the values of SP, DP, TCH, TG, HDL-CH, LDL-CH, EM-CH, BMI, DCSC, DPAC, age and IHV of the SIH patients, and that Y = -12.4583 + 0.1127SP -1.1977EM-CH + 0.9788LDL-CH + 0.2477BMI + 0.0382DCSC + 0.0248DP, P < 0.0001 approximately 0.05. CONCLUSIONS: The findings in the present study suggest that significantly increased systolic and diastolic pressure, low density lipoprotein cholesterol, body mass index and daily cigarette smoking consumption, and significantly decreased erythrocytic membrane cholesterol may be likely the main factors affecting intracerebral hemorrhagic volumes in patients with acute spontaneous intracerebral hemorrhage.