Erythropoietin attenuates cerebral ischemia-induced cognitive dysfunction through stimulation of hippocampal CREB phosphorylation in C57BL/6 mice

AIM: To investigate the neuroprotective effect of erythropoietin (EPO) on cognitive dysfunction and neuronal injury in hippocampal CA1 region induced by cerebral ischemia in mice. METHODS: Male C57BL/6 green fluorescent protein-transgenic mice were randomly divided into 3 groups: sham operation group (sham), ischemia/reperfusion group (I/R) and EPO-treated group. Transient cerebral global ischemia was induced by bilateral common carotid artery occlusion (2-VO). The step-down test was used to measure the capacity of learning and memory of the animals in each group. Nissl staining was applied to examine the neuronal number in hippocampal CA1 region. The expression of phosphorylated cAMP response element-binding protein (pCREB) was determined by Western blotting. Alterations of dendritic morphology in hippocampal CA1 region were evaluated using laser scanning confocal microscopy and Neurolucida software. RESULTS: Transient cerebral ischemia caused deficits of spatial learning and memory, and delayed neuronal death and loss of dendritic spines in hippocampal CA1 region were also obvious. The EPO treatment significantly improved the cognitive function in cerebral ischemic mice, and the protein expression of pCREB was obviously increased. At the same time, neuronal death and loss of dendritic spines were reduced in hippocampal CA1 region. CONCLUSION: Erythropoietin increases the protein expression of pCREB, and reduces neuronal death and loss of dendritic spines. These processes may be responsible for erythropoietin-mediated neuroprotective effects and the improvement of cognitive function in cerebral ischemic mice.     

梭梭14-3-3蛋白基因HaFT-9的克隆及特性分析

【目的】分析梭梭14-3-3蛋白基因HaFT-9在不同胁迫下的表达模式,解析HaFT-9蛋白结合特性,为研究梭梭抗逆分子机制提供理论基础。【方法】以梭梭cDNA为模板克隆14-3-3蛋白基因HaFT-9,利用qRT-PCR技术对该基因在不同胁迫下的表达模式进行分析,并通过酵母双杂交实验验证HaFT-9蛋白结合特性。【结果】克隆得到ORF长度为789 bp的梭梭14-3-3蛋白基因,命名为HaFT-9。梭梭幼苗在20% PEG6000、200 mmol/L NaCl和100 μmol/L ABA处理下,HaFT-9的表达量均在胁迫6 h时达到峰值,在20 μmol/L IAA处理下,HaFT-9的表达量在胁迫1 h时达到峰值。梭梭幼苗经40、55℃和55、65℃模拟地表高温胁迫,高温胁迫适应性试验中的常温复原阶段HaFT-9均上调表达。酵母双杂交实验表明HaFT-9可与自身形成同源二聚体。【结论】克隆得到HaFT-9基因(登录号为API85525.1),该基因可响应干旱胁迫、盐胁迫、ABA和IAA处理,并与高温胁迫适应性相关,HaFT-9蛋白能与自身互作形成同源二聚体。     

半胱胺对绒山羊瘤胃及盲肠内环境的影响

本试验选用4头6月龄、平均体重为16kg左右、安装永久性瘤胃瘘管和盲肠瘘管的辽宁绒山羊,平均分为半胱胺组和对照组,观察辽宁绒山羊瘤胃及盲肠内环境的变化。结果表明:半胱胺组瘤胃pH较对照组差异不显著(P0.05)。半胱胺组瘤胃NH3-N浓度、瘤胃蛋白浓度较对照组差异显著(P0.05);盲肠pH、蛋白浓度半胱胺组较对照组均差异不显著(P0.05)。本试验结果提示,添加半胱胺后可改变瘤胃内环境,有利于瘤胃微生物蛋白的合成。     

The influences of ambient temperature and crude protein levels on performance and serum biochemical parameters in broilers

This study was undertaken to investigate the effects of ambient temperature, crude protein levels and their interaction on performance and serum biochemical parameters of broiler chickens. A total of 216 Arbor Acre broiler chickens (108 males and 108 females) were used in a 2 × 3 factorial arrangement and randomly reared at two temperatures (normal temperature: 23 °C; daily cyclic high temperature: 28–32 °C) and fed on three diets with different crude protein levels (153.3, 183.3 or 213.3 g/kg, with constant essential amino acids) from 28 to 42 days of age. Daily cyclic high ambient temperature decreased final body weight, average daily weight gain, average daily feed intake and serum total protein contents (p < 0.001, p < 0.001, p < 0.001, p = 0.008 respectively), but increased feed/gain, mortality, respiratory rate, rectal temperature, serum uric acid contents and serum creatine kinase activity (p = 0.008, p = 0.003, p < 0.0001, p < 0.0001, p < 0.0001, p = 0.003 respectively), irrespective of crude protein levels. At the ambient temperature, reducing crude protein levels resulted in an increase in feed/gain (p < 0.001), but a decrease in serum total protein and uric acid contents. Only serum creatine kinase activity in broiler chickens was interacted by daily cyclic high ambient temperature and dietary crude protein levels (p = 0.003). These results indicated that daily cyclic high ambient temperature had a great effect on performance and serum biochemical parameters in broiler chickens, whereas dietary crude protein levels affected them partially.     

甘薯可溶性蛋白、叶绿素及ATP含量变化与品种抗旱性关系的研究

 对水分胁迫下甘薯叶片部分物质和能量代谢指标的研究结果表明 ,水分胁迫下叶片中可溶性蛋白含量明显增加 ;叶绿素a、叶绿素b、总叶绿素含量及叶绿素a/b比值与对照相比均有所下降 ;ATP含量有增有减 ,但品种抗旱性愈强 ,ATP含量愈高。叶片中可溶性蛋白含量、叶绿素a/b比值、ATP含量占对照百分率与品种抗旱性间的相关系数r分别为 0 .896 8、- 0 .85 0 9和 0 .82 0 0 ,P <0 .0 1。因此 ,这些指标可用于甘薯不同品种抗旱性的评定     

稻瘟菌基因组规模分泌蛋白的预测分析

利用信号肽分析软件SignalP v3．0，亚细胞器蛋白定位软件TargetP v1．01，GPI-锚定位点软件big—PI Predictor和跨膜螺旋结构软件THMM-2．0这4个软件分析预测稻瘟菌12595个蛋白序列中有1134个分泌蛋白。编码这些蛋白的可读框（ORF）最小值为78bp，最大值为7849bp，平均1231bp。引导它们的信号肽长度介于15—45个氨基酸之间，平均为21个氨基酸。435个分泌蛋白具有功能描述，主要是与代谢有关的酶类。分析了其中降解细胞壁组分和与致病相关的酶类，提示在稻瘟菌侵染水稻不同阶段产生的关键酶及基因的功能需进一步研究。     

小麦热激蛋白转录因子C家族基因的结构及其在干旱胁迫中的表达

小麦热激蛋白转录因子在热胁迫和耐热性产生的过程中发挥重要作用。为进一步了解小麦热激蛋白转录因子C亚家族,本研究采用全基因组信息保守结构域比对和进化聚类分析,共鉴定了24个小麦TaHsfC基因,并对TaHsfC亚家族成员染色体定位、基因结构、亚细胞定位、对应蛋白质的氨基酸特点、基因表达进行了分析。结果表明,24个TaHsfCs主要分布在5条染色体上,其中A基因组中有7个,B基因组中9个,D基因组中有6个,另外两个所在染色体位置不清楚。TaHsfCs含有0～2个内含子,其中 TaHsfC1亚族基因含有1～2个内含子, TaHsfC2亚族基因则含有0～1个内含子。聚类分析表明,小麦TaHsfC成员共分为 TaHsfC1和 TaHsfC2两个亚族。24个TaHsfCs成员全部定位于细胞核。在15%PEG模拟干旱条件下,干旱敏感品种矮抗58和耐旱品种晋麦47中有10个TaHsfC基因显著上调表达,其中 TaHsfC2亚族基因的表达量上调倍数均高于 TaHsfC1亚族基因(其中 TaHsfC2j和 TaHsfC2k未在晋麦47中检测到表达量)。本研究可为探索小麦热激蛋白转录因子C家族基因在小麦抗旱中的分子机制提供一定的理论支撑。     

The Distribution of Soluble Copper- and Zinc-Binding Proteins in Liver and Kidney of Chronic Copper-Poisoned Goats

The distribution of copper and zinc among the soluble proteins in the liver and kidney from chronic copper-poisoned goats was examined after gel filtration of the proteins. The concentrations of copper in the liver and kidney cortex from five experimentally copper-poisoned goats were: 550–810 µg/g liver and 190–420 µg/g kidney cortex (wet weight). In general the copper-binding proteins from, both the liver and kidney samples were separated into two different fractions with approximate molecular weights (m.w.) of > 65,000 and 10,000, respectively. From the liver samples, varying amounts of copper were eluted in a fourth fraction with m.w. < 2,000. In the majority of kidney samples the dominating copper-binding protein fraction was the high molecular weight fraction. Absolute amounts of copper recovered in the metallothionein-like protein fraction were nearly the samt for all samples investigated. The distribution of zinc-binding proteins in both liver and kidney samples was nearly the same. The high molecular weight fraction dominated, and no zinc was bound to metallothionein-like proteins.     

小麦白粉菌ADP/ATP载体蛋白基因的克隆及表达特征分析

 小麦白粉病是小麦上的一种重要病害。研究小麦白粉菌致病相关基因及其在致病过程中的作用,对于丰富小麦白粉菌致病的分子机制和筛选防治新靶标具有重要意义。前期转录组测序结果提示一个小麦白粉菌ADP/ATP载体蛋白（ADP/ATP carrier protein,AACP）在小麦与白粉菌互作时上调表达。本研究利用cDNA末端快速克隆技术（rapid\|amplification of cDNA ends,RACE）获得了长945 bp具有完整开放阅读框的小麦白粉菌ADP/ATP carrier基因序列,暂命名为BgtAACPx（GenBank登录号为MF197857）该基因编码314个氨基酸残基,利用相关软件进行生物信息分析,结果表明该蛋白为疏水性,含有6个跨膜区,亚细胞定位在线粒体上。与已有小麦白粉菌AACP蛋白的同源性为97%,是一个新的蛋白,同时构建了与其同源蛋白的遗传进化树。定量结果表明该基因在小麦白粉菌吸器期（48 h）、次生菌丝（72 h）至白粉菌刚产孢子又未形成孢子堆（120 h）这段时期高表达,提示该基因可能参与小麦白粉菌对小麦的致病过程。     

Soy protein concentrate as a substitute for fish meal in diets for juvenile Acanthopagrus schlegelii: effects on growth,phosphorus discharge and digestive enzyme activity

The current high demand and cost of fish meal (FM) necessities the evaluation of alternative plant protein ingredients in diets of farmed marine fish. A 56‐day feeding trial was performed to study the effects of replacement of FM with soy protein concentrate (SPC) in diets of Acanthopagrus schlegelii. Diets were prepared at levels of 0%, 30%, 47.5%, 65%, 82.5% and 100% SPC, respectively, replacing FM. The results indicated no significant differences (p > .05) in % weight gain (WG) and specific growth rate of fish fed S30% to S82.5% diets compared with the control diet while, further inclusion at 100% significantly depressed growth performance. SPC inclusion and phytase supplementation significantly affected the phosphorous discharge (P‐load) showing lowest value (3.83 ± 0.53 g/kg WG) in S100% compared to control (14.79 ± 0.37 g/kg WG) and in fish fed S30% diet (13.24 ± 0.89 g/kg WG) (p < .05). The results of this study showed that FM could be substituted up to 82.5% by SPC in the diet of Acanthopagrus schlegelii fingerlings (5.53 ± 0.12 g) without any adverse effects. Phytase supplementation SPC based diets could be effective in reducing the phosphorus load in the aquatic culture environment.