Effect of miRNA-33 on ABCA1/ABCG1 expression in RAW264.7 macrophage-derived foam cells after Hcy treatment

AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model. Oil red O staining was used to determine whether the model was established successfully. miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h. The intracellular lipid droplets were observed by Oil red O staining. The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot. The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05). The intracellular cholesterol content was increased gradually (P<0.05), and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group. Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05). No diffe-rence of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells.     

Effect of 4-chloro-2-methylphenoxyacetic acid and 2,4-dimethylphenol on human erythrocytes

The effects of exposure of human erythrocytes to different concentrations of 4-chloro-2-methylphenoxyacetic acid (MCPA) and its metabolite—2,4-dimethylphenol (2,4-DMP) were studied. The investigations concerned mainly the content of glutathione (GSH and GSSG), glutathione peroxidase (GSH-Px), glutathione transferase (GST), and the level of adenine energy charge (AEC). Reactive oxygen species (ROS) such as hydroxyl radical, superoxide anion, hydrogen peroxide, and nitric oxide are produced during normal processes in the cell. Under normal conditions, antioxidant systems of the cell minimize damage caused by ROS. When ROS generation increases to an extent that it overcomes the cellular antioxidant systems, the result is oxidative stress. We observed that MCPA and 2,4-DMP decreased the level of GSH in erythrocytes in comparison with control. MCPA did not affect glutathione peroxidase and glutathione transferase activity, while 2,4-DMP increased their activity. 2,4-DMP decreased the level of ATP and increased the content of ADP and AMP, leading to the fall of the level of AEC. MCPA and 2,4-DMP transform hemoglobin into methemoglobin, thus preventing oxygen transport. Comparison of the toxicity of MCPA and 2,4-DMP revealed that the most prominent changes occurred in human erythrocytes incubated with 2,4-DMP.     

Guanidinoacetic acid supplementation in broiler chickens fed on corn-soybean diets affects performance in the finisher period and energy metabolites in breast muscle independent of diet nutrient density

(1) Guanidinoacetic acid (GAA) is the single immediate endogenous precursor of creatine (Cr). It was hypothesised that dietary GAA would have different effects on performance and energy metabolites in breast muscle depending on the nutrient density (ND) of corn-soybean-based diets.

(2) A total of 540 one-day-old male Ross 308 broilers were allocated to 9 dietary treatments with 6 replicates (10 birds each) in a 3 × 3 factorial arrangement with three levels of ND (low, 2800; medium, 2950 and high, 3100 kcal metabolizable energy (ME)/kg; and with the other nutrients being constant relative to ME) and supplemented with three levels of GAA (0, 0.6 and 1.2 g/kg) in a 42-d feeding trial.

(3) In the starter and grower periods, increasing levels of ND improved body weight (BW), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR), with the exception of ADFI in the starter period. GAA supplementation did not affect performance characteristics. All performance indicators responded markedly to increasing ND in the finisher period, whereas the highest GAA level reduced ADFI compared to the unsupplemented control (156 vs. 162 g/d) and concomitantly FCR (1.81 vs. 1.93). No interactive effects were noted for any performance trait. The high ND diet resulted in more breast meat yield on d42, associated with higher fat content and darker colour compared to the other ND levels. The GAA supplementation did not affect carcass and breast traits. At the end of the experiment, Cr was elevated when feeding GAA at 1.2 g/kg (5455 vs. 4338 mg/kg fresh muscle).

(4) To conclude, ND had a substantial effect on performance and carcass traits, whereas any effect of GAA was limited to FCR in the finisher period and independent of diet ND level.     

Lipid-rich bovine serum albumin improves the viability and hatching ability of porcine blastocysts produced in vitro

The effects of lipid-rich bovine serum albumin (LR-BSA) on the development of porcine blastocysts produced in vitro were examined. Addition of 0.5 to 5 mg/ml LR-BSA to porcine blastocyst medium (PBM) from Day 5 (Day 0 = in vitro fertilization) significantly increased the hatching rates of blastocysts on Day 7 and the total cell numbers in Day-7 blastocysts. When Day-5 blastocysts were cultured with PBM alone, PBM containing LR-BSA, recombinant human serum albumin or fatty acid-free BSA, addition of LR-BSA significantly enhanced hatching rates and the cell number in blastocysts that survived compared with other treatments. The diameter, ATP content and numbers of both inner cell mass and total cells in Day-6 and Day-7 blastocysts cultured with PBM containing LR-BSA were significantly higher than in blastocysts cultured with PBM alone, whereas LR-BSA had no effect on mitochondrial membrane potential. The mRNA levels of enzymes involved in fatty acid metabolism and β-oxidation (ACSL1, ACSL3, CPT1, CPT2 and KAT) in Day-7 blastocysts were significantly upregulated by the addition of LR-BSA. The results indicated that LR-BSA enhanced hatching ability and quality of porcine blastocysts produced in vitro, as determined by ATP content, blastocyst diameter and expression levels of the specific genes, suggesting that the stimulatory effects of LR-BSA arise from lipids bound to albumin.     

土壤微生物膜对风沙土固沙保水特性的影响

土壤微生物膜胞外聚合物具有较强的黏结性,为增强风沙土抗蚀性,提高风沙土保水性提供了新途径。鉴于此,该研究采用室内控制试验设置6个微生物菌剂施用水平(0、1、3、5、7和10 g/kg),以期阐明土壤微生物膜对风沙土特性的影响。研究结果表明:1)施用微生物菌剂可产生土壤微生物膜,并胶结风沙土颗粒。2)试验结束时(第49 d),与对照组相比,不同菌剂施用处理的土壤容重降低0.54%～8.88%,孔隙度提高0.39%～3.91%,含水率显著提高0.11%～0.25%(P0.05),大团聚体质量分数增加5.59%～25.01%,土壤pH值由7.16显著增加至8.32～9.23(P0.05)。3)土壤多糖含量对土壤特性的解释率为47%,是影响土壤特性的关键因素。土壤微生物膜能够有效改善风沙土特性,增强风沙土抗蚀性,提高风沙土保水性,但较高的微生物菌剂施用量会增加土壤pH值,造成土壤盐碱化,建议微生物菌剂施用水平为1～5 g/kg。研究成果有利于深入理解微生物膜固沙保水机制,并可为沙化土地防治新技术研发提供重要理论依据和科技支撑。     

Purinergic control of the quail rectum: modulation of adenosine 5'-triphosphate-mediated contraction with acetylcholine

Electrical field stimulation (EFS) induces frequency-dependent contractions of the longitudinal muscle of isolated quail rectum which were sensitive to tetrodotoxin. The aim of the present study was to investigate whether purinergic neurons are implicated in the response to nerve stimulation. The shape of the EFS-induced contractile response was different depending on stimulus frequency; low frequencies (0.5-2 Hz) induced fast monophasic contractions with a small subsequent relaxation; whereas higher frequencies (5-50 Hz) induced biphasic contractile response that comprised fast initial component (as in case of low frequency) and a slow delayed contractile component in addition to the relaxation that follows the fast contractile component. Prior application of atropine (10 microM) completely abolished the slow delayed component but significantly enhanced the fast initial contractile component. Physostigmine (1-10 microM) significantly enhanced the slow delayed component with an inhibitory effect on the initial fast component. The nonspecific purinergic receptor antagonist, suramin (100-500 microM) significantly inhibited the fast initial contractile component with no significant effect on the slow delayed one. Complete blockade of the fast component was achieved by prior application of a combination consisted of suramin (50 microM) and pyridoxicalphosphate-6-azophenyl 2',4'-disulphonic acid tetrasodium (PPADS; 10 microM). Exogenous applications of adenosine 5'-triphosphate and acetylcholine (10 microM each), produced contractile responses that mimicked those induced by EFS. These data suggest that ATP is the main noncholinergic excitatory transmitter controlling the contractile activity of the quail rectum; and that its action could be modulated by acetylcholine.     

胞外多聚物在污泥颗粒化过程中的作用研究

本文研究了胞外多聚物(ECP)在污泥颗粒化过程中的作用。试验采用小试UASB反应器,以人工葡萄糖配水作为进料。测定结果表明,在颗粒污泥培养过程中,随着颗粒污泥的形成、生长和成熟,污泥ECP含量逐渐下降,从开始的6.36减小到30mg/gVSS。当反应器酸化、污泥性能变差或运行不稳定时,ECP含量则表现为急剧上升,达70mg/gVSS。这些结果说明污泥颗粒化过程中ECP所起的作用不大。对有关ECP的影响及作用机理,以及污泥颗粒化机制,文章进行了分析讨论。