Effect of mild hypothermia on energy metabolism and hydroxyl radical production after cerebral ischemia/reperfusion in gerbils

AIM: To study the effect of mild hypothermia on energy metabolism and hydroxyl radical production as well as delayed neuronal death (DND) in hippocampus during cerebral ischemia/reperfusion in gerbils.METHODS: Forebrain ischemia was induced by occluding bilateral common carotid arteries with aneurysm clamps for 10 min in gerbils. The DND in hippocampal CA1 sector was assessed by histological examination, and hydroxyl radical, ATP (adenosine triphosphate), ADP (adenosine diphosphate),AMP (adenosine monophosphate) levels were determined by high performance liquid chromatography with electrochemical or ultraviolet detection. RESULTS: The number of survival neuronal in hippocampal CA1 sector in mild hypothermia + I/R group was more than that in I/R group after ischemia/reperfusion 96 h. The content of 2,3-DHBA (2,3- dihydroxybenzoic acid) in hippocampus in I/R group was much higher than those in sham operation and mild hypothermia + I/R group after reperfusion 6 h (P＜0.01), but there were no significant differences in 2,3-DHBA outputs among 3 groups 48 h and 96 h after reperfusion. There were no obvious differences in ATP, ADP, AMP level in hippocampus among 3 groups 6 h after reperfusion. However, the content of ATP,ADP,AMP in mild hypothermia + I/R group was much higher than those in I/R group 48 and 96 h after reperfusion (P＜0.01).CONCLUSION: Mild hypothermia can reduced DND by improving the cerebral energy metabolism during forebrain ischemia/reperfusion in gerbils.     

组合填料对水质、罗非鱼生长及水体微生物群落功能多样性的影响

采用组合填料(全塑性夹片和维纶醛化丝)作为生物膜载体,以土著微生物为生物膜菌源,研究其对水质、罗非鱼生长及水体微生物群落功能多样性的影响。结果表明:组合填料可促进水体中NO_2~-、DO、IC含量显著升高,使水体中pH、NO_3~-含量显著下降,同时有使水体中NH_4~+、TOC含量下降的趋势;和对照组相比,组合填料组罗非鱼增重量显著升高,饵料系数显著降低,填料悬挂密度越高,增重效果越明显;组合填料能显著降低水体细菌数量,而对水体微生物多样性无显著影响;组合填料明显提高了水体微生物对糖类、醇类、酯类的利用率,降低了水体微生物对胺类和酸类的利用率。RDA分析表明,组合填料组和对照组之间的水体微生物代谢多样性存在一定的差异,而DO、pH和硝酸盐是影响处理组和对照组水体微生物代谢多样性差异的主要环境因子,显示组合填料在一定程度上可有效改善水质,促进罗非鱼生长,降低水体细菌数量,影响水体微生物群落功能多样性。     

Physiological and biochemical parameters for egg quality determination in lake trout,Salmo trutta lacustris

The present study investigated the relationships between egg viability and ovarian fluid composition, egg physiology and egg metabolism in lake trout, Salmo trutta lacustris, to obtain biomarkers for egg quality determination. The ovarian fluid pH, protein levels and activities of aspartate aminotransferase and -d-glucuronidase were significantly correlated with egg viability expressed as the number of eyed stage embryos. Regression models demonstrated that an ovarian fluid pH between 8.44 and 8.57, protein levels below 235.56 mg 100 ml^–1ovarian fluid, aspartate aminotransferase activity below 31.65 m min^–1 l^–1ovarian fluid and -d-glucuronidase activity below 8.62 m min^–1 l^–1 ovarian fluid characterized egg batches with high viability (80%).The increase in the egg wet weight during water hardening was also significantly correlated with the number of eyed stage embryos, and egg batches with high egg viability (80%) increased in wet weight by 13% during water hardening.From the investigated metabolic parameters the number of eyed stage embryos was significantly correlated with activities of NADP-dependent isocitrate dehydrogenase (egg viability 80% at 2.07 nM min^–1 mg^–1 protein) and NAD-dependent malate dehydrogenase (egg viability 80% at 47.25 nM min^–1 mg^–1 protein), with the respiration rate (egg viability 80% at 8.71 nM min^–1 mg^–1 protein), with the ratio of NADH to NAD levels (egg viability 80% 0.872), with the levels of free, non-esterified fatty acids (egg viability 80% 72.34 g mg^–1 protein), and the ratio of non esterified to esterified fatty acids (egg viability 80% at 0.749). Also, subjective and visual control methods were described to distinguish between batches with viable and non viable eggs.     

Are Primary Cultures of Trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) Ventricular Cardiomyocytes Metabolically Viable?

The objectives of this study were to establish a protocol for the isolation of metabolically viable ventricular cardiomyocytes from the rainbow trout and to determine which measures may best reflect viability 24 h after isolation. Cardiomyocytes were isolated by enzymatic digestion and maintained in cell suspension. Viability was assessed using Trypan blue dye exclusion, ATP content, oxygen consumption and lactate dehydrogenase (LDH) leakage into the medium. Viability, assessed by these measures did not significantly change over the time period of this study. ATP content did correlate significantly with oxygen consumption but not with Trypan blue exclusion. We conclude that primary cultured cardiomyocytes remain metabolically viable for at least 24 h after isolation. Also, it appears that ATP content and oxygen consumption most adequately reflect metabolic cell viability. To be confident with a culture, however, a combination of viability measures is necessary when isolating cardiomyocytes from fish.     

复合益生菌对眼斑双锯鱼(Amphiprion ocellaris)肠道消化酶、菌群结构以及形态的影响

为探讨饲料中添加复合益生菌对眼斑双锯鱼(Amphiprion ocellaris)肠道消化酶、菌群结构及形态的影响,将135尾初始体重(0.79±0.01)g的眼斑双锯鱼随机分为3组,每组3个重复,分别投喂复合益生菌有效活菌添加量为0%(对照组)、3%(L3组)和6%(L6组)的饲料,进行为期28 d的养殖试验。结果显示,复合益生菌能提升试验鱼肠道蛋白酶的活性,其中L3组肠道蛋白酶显著提升(P<0.05);能显著降低试验鱼肠道淀粉酶活性(P<0.05);能降低脂肪酶活性(P>0.05)。随着饲料中益生菌添加量的增加,试验鱼肠道中变形菌门丰度开始增多,拟杆菌门丰度降低。饲料中益生菌添加量的增加显著降低试验鱼肠道肌层厚度并显著增加试验鱼肠道绒毛高度(P<0.05)。可见,饲料中添加3%复合益生菌可改善眼斑双锯鱼肠道消化酶活性,提高肠道微生物群落多样性,改善肠道组织形态和功能。     

高温和低氧胁迫对两种规格刺参半致死时间及生理机能的影响

通过实验生态学方法,针对2种规格的刺参(Apostichopus japonicus)(大:28.00~36.00 g;小:9.00~13.00 g),测定温度为(25.0±0.5)℃溶解氧(DO)水平为(1.0±0.1)mg/L条件下刺参的半致死时间(LT_(50));比较分析了DO水平分别为(1.0±0.1)mg/L、(3.0±0.1)mg/L及6.5 mg/L(正常DO含量)条件下2种规格刺参的昼夜代谢水平;监测了DO水平分别为(1.0±0.1)mg/L和(3.0±0.1)mg/L时低氧胁迫24 h及复氧72 h期间,2种规格刺参体腔液和呼吸树中的谷胱甘肽(GSH)含量、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性及体腔液中皮质醇水平的变化,以探究夏季高温期(25.0±0.5)℃低氧胁迫对不同规格刺参半致死时间及生理机能的影响。结果显示:当DO浓度为1 mg/L时,大规格刺参半致死时间(LT_(50))为33.37 h,小规格刺参为28.84 h,2种规格刺参的代谢水平显著低于常氧对照组,夜间代谢强度高于白天,且小规格刺参的代谢强度高于大规格刺参。1 mg/L低氧胁迫期间,大规格刺参体腔液及2种规格刺参呼吸树中上述4种抗氧化指标的变化趋势大致相同,与对照组相比,随着低氧暴露时间的延长,GSH含量下降,SOD和T-AOC活力降低,CAT活力升高;然而胁迫结束时其GSH含量、CAT、SOD和T-AOC活力均与对照组无显著差异(P0.05)。解除胁迫复氧72 h后,2种规格刺参体腔液的4种抗氧化指标均恢复到对照组水平;而呼吸树的这4种指标则未完全恢复。3 mg/L低氧胁迫期间,2种规格刺参体腔液和呼吸树的GSH含量、CAT、SOD和T-AOC活力的变化趋势与1 mg/L组大体一致,复氧72 h后体腔液的各项指标恢复到对照组水平;除T-AOC,呼吸树的其余3种指标亦完全恢复。低氧胁迫后小规格刺参的恢复能力高于大规格刺参。在DO为1 mg/L和3 mg/L条件下,胁迫结束时2种规格刺参体腔液中的皮质醇含量显著高于对照组(P0.05),且大规格刺参皮质醇含量高于小规格,复氧后二者均恢复到对照水平。结果表明,低氧胁迫持续时间不超过其半致死时间(本研究以24h为例),刺参可通过自身调节减轻机体的氧化损伤;一旦超出半致死时间,将产生不可逆损伤,最终导致死亡。     

Study of the Protective Effects in PEPC Transgenic Rice

The diurnal course of chlorophyll fluorescence parameter and active oxygen metabolism of flag leaves in PEPC transgenic and untransformed rice Kitaake were studied. The results showed that the photosynthetic rate under high light intensity has been increased by 50% and photoinhibition of photosynthesis in PEPC transgenic was alleviated after the introduction of PEPC gene from maize into rice. It was demonstrated that the increment of photosynthesis in PEPC transgenic was related to the introduction of PEPC gene using specific inhibitor of PEPC. Photoinhibition of photosynthesis in different genotypes exists at noon under natural condition. PEPC transgenic rice exhibited a less decrease in Fv/Fm, a less photoinhibition and a higher efficiency of light energy conversion to chemical energy and lower thermal energy dissipation.These results provided the physiological basis on the mechanism of tolerance to photoinhibition and rice breeding with high photosynthetic efficiency.     

壳聚糖诱导脐橙果实抗病性、水杨酸及活性氧代谢变化

【目的】探讨壳聚糖处理对脐橙果实抗病性的诱导效果,以及对果实内源水杨酸(SA)含量和活性氧代谢的影响。【方法】以脐橙果实为试材,采收后不同时间用2%壳聚糖溶液浸泡处理1min,测定壳聚糖处理对果实青霉病抗病性、水杨酸含量和活性氧代谢的影响。【结果】壳聚糖处理显著降低脐橙果实接种发病率的升高和病斑直径的扩展(P0.01),诱导2d时抗病效果最显著。2%壳聚糖处理果实果皮内游离态SA和过氧化氢(H2O2)含量明显高于对照果实;壳聚糖处理提高脐橙果实过氧化物酶(POD)、谷胱甘肽还原酶(GR)活性,抑制过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性,延缓果实还原型抗坏血酸(AsA)含量的下降,引起诱导初期还原型谷胱甘肽(GSH)的积累。【结论】2%壳聚糖处理可以诱导脐橙果实对青霉病的抗病性,其诱导的脐橙果实抗病反应涉及果皮内SA含量和活性氧水平的调节以及抗氧化酶活性的变化。     

Improvement of flesh quality in rainbow trout (Oncorhynchus mykiss) fed supranutritional dietary selenium yeast is associated with the inhibited muscle protein degradation

Supranutritional dietary selenium (Se) has been demonstrated beneficial for fish health, however, its effects on fish flesh quality remain unclear. This study investigated the effects of supranutritional dietary Se on the flesh quality of rainbow trout (Oncorhynchus mykiss) and their potential mechanism. Fish were fed a basal diet supplemented with or without graded Se yeast for 10 weeks. Results showed that Se supplementation significantly increased fillet crude protein and enhanced fillet water‐holding capacity as well as fillet firmness. Supplementing with both 2 and 4 mg/kg Se significantly downregulated the expressions of two autophagy–lysosome‐related genes (autophagy‐related 12‐like and gamma‐aminobutyric acid type A receptor‐associated protein‐like 1) in fish muscle, while supplementing with 4 mg/kg Se also significantly downregulated the expression of two ubiquitin–proteasome‐related genes (muscle RING finger 2 and F‐box protein 25). Correlation analysis indicated that the improved fillet quality parameters were closely correlated with the expressions of these differentially expressed genes. This study revealed that dietary Se was effective for the improvement of rainbow trout flesh quality, and the improved fish flesh quality was associated with the inhibited protein degradation in fish muscle.     

幼建鲤磷缺乏的组织病理观察

为研究饲料不同磷水平对吉富罗非鱼营养代谢及肠道微生物的影响,实验以磷酸二氢钙作为磷源,配制成总磷含量为0.26%(低磷组)、0.81%(适磷组)和1.51%(高磷组)的3组等氮等能饲料,每种饲料为一个处理,每个处理设置4个重复,每个重复30尾鱼,用3种饲料分别饲喂初始体重为(8.42±0.09) g的吉富罗非鱼8周。结果显示,适磷组吉富罗非鱼的增重率和特定生长率显著高于低磷组和高磷组,饲料系数在适磷组最低。脏体比、肝体比和肥满度随饲料磷水平的升高呈逐渐下降趋势。随着饲料磷水平的增加,吉富罗非鱼干物质、粗蛋白、粗脂肪、磷的表观消化率显著降低。对筛选到的差异代谢物进行KEGG注释和富集分析发现,饲料磷缺乏时下调的差异代谢物主要富集的代谢通路为氰胺酸代谢、葡萄糖酸酯的生物合成、丙氨酸、天冬氨酸和谷氨酸代谢,上调的差异代谢物主要富集的代谢通路为脂肪酸合成;当饲料磷过量时下调的差异代谢物主要富集代谢通路为精氨酸和脯氨酸代谢、苯丙酸的生物合成,上调的差异代谢物主要富集代谢通路为氨基糖和核苷酸糖的代谢。Ace、Chao1和Shannon指数表明,吉富罗非鱼肠道菌群丰度和多样性随着饲料磷水平的升高呈现升高趋势。厚壁菌门、放线菌门、变形菌门和拟杆菌门为吉富罗非鱼肠道菌群中的优势菌门。罗姆布茨菌属、鲸杆菌属等有益菌属的丰度在适磷组最高,分枝杆菌属和拟杆菌属的丰度随饲料磷含量的增加呈下降趋势。本研究表明,饲料中适量的磷能提高吉富罗非鱼对饲料的表观消化率,磷缺乏和过量会抑制吉富罗非鱼的氨基酸代谢,磷缺乏会加快脂肪酸合成进程;饲料中适量磷可以提高菌群丰度和多样性,有利于肠道健康。