天麻糖复合物抗衰老作用的实验研究

天麻糖复合物是从兰科植物天麻的根茎中提取的水溶性物质,经柱色谱将其分为两个组分,分别命名为：Con.1,Con.2;考察灌胃后两个组分对小鼠血清中超氧化物岐化酶活性和过氧化脂质含量的影响,同时探讨两个组分对小鼠和果蝇的抗衰老作用及其机制.结果Con.1 0.1g/kg.d能明显提高小鼠血清中T-SOD活性,降低血清中过氧化脂质降解产物（MDA）含量;Con.1可延长果蝇的平均寿命和最高寿命,因此Con.1具有明显的抗衰老作用。但Con.2影响结果不明量.     

产1,3-丙二醇新型重组大肠杆菌的构建

利用PCR技术从大肠杆菌(Escherichia coli )中扩增出1.16 kb的编码1,3-丙二醇氧化还原酶同工酶的基因yqhD,将其连接到表达载体pEtac,得到重组载体pEtac-yqhD,重组载体在大肠杆菌JM109中得到高效表达。SDS_PAGE分析显示融合表达产物的分子量均为43 kD,同核酸序列测定所推导的值相符。对含有yqh-D的基因工程菌进行表达研究表明：37 ℃,以1.0 mmol /L IPTG诱导4 h,1,3-丙二醇氧化还原酶同工酶的酶活力达到120 u/mg蛋白,而对照菌株的酶活力为0.5 u/mg蛋白。再将含甘油脱水酶基因dhaB和含1,3-丙二醇氧化还原酶同工酶基因yqhD的重组质粒共转化大肠杆菌JM109得到重组大肠杆菌JM109（pUCtac-dhaB, pEtac-yqhD）,该菌株在好氧条件下,以1.0mmol/L IPTG诱导可将50 g/L甘油转化为38.0 g/L 1,3-丙二醇。首次发现1,3-丙二醇氧化还原酶同工酶在好氧条件下表现出较高的活性。     

四倍体长穗偃麦草(Agropyron elongatum 4x)染色体组构成的生化和SSR标记分析

应用等电聚焦(IEF)和SDS PAGE方法分析了二倍体长穗偃麦草(Agropyronelongatum2x)和四倍体长穗偃麦草(Ag.elongatum4x)的16种同工酶和3种贮藏蛋白的电泳图谱。结果表明,只有两种同工酶在四倍体和二倍体长穗偃麦草之间表现相同的酶谱,该类型仅占分析标记总数的10.5%;而10种同工酶和3种贮藏蛋白的电泳图谱在四倍体中除了具有全部二倍体的谱带以外,还有自己独特的条带,该类型最多,占分析标记总数的63.2%;另外5种同工酶在二倍体和四倍体之间只有部分条带相同,同时具有各自特异的条带,该类型占分析标记总数的26.3%。由此推测,四倍体长穗偃麦草可能是一个异源四倍体,即只含有一个起源于二倍体类型的染色体组Ee,而另一个染色体组在所分析的生化标记上明显不同于近缘种中的St、J和N染色体组,其起源尚待进一步研究。进一步用小麦的SSR引物对二倍体和四倍体长穗偃麦草进行扩增,结果表明大多数SSR引物在四倍体中既能扩出与二倍体相同的条带,同时还有其特异的条带,这一结果验证了由生化标记得出的四倍体长穗偃麦草是异源四倍体的初步结论。     

Synthesis and evaluation of novel N–H and N-substituted indole-2- and 3-carboxamide derivatives as antioxidants agents

We have previously reported on the synthesis of novel indole derivatives where some compounds showed significant antioxidant activity. Here, we report the synthesis of novel N–H and N-substituted indole-2- and 3-carboxamide derivatives and investigated their antioxidant role in order to identify structural characteristics responsible for activity. Although all compounds showed a strong inhibitory (95–100%) effect on superoxide anion (SOD) only compounds 4, 5 and 6 showed simliar potency for the inhibition of lipid peroxidation (81–94%) which revealed that compounds 4, 5 and 6 possessed highly potent antioxidant properties. Substitution in the 1-position of the indole ring caused the significant differences between the activity results regarding lipid peroxidation inhibition.     

Acidotoxicity and acid-sensing ion channels contribute to motoneuron degeneration

Amyotrophic lateral sclerosis (ALS) is a fatal neurological condition with no cure. Mitochondrial dysfunction, Ca²⁺ overloading and local hypoxic/ischemic environments have been implicated in the pathophysiology of ALS and are conditions that may initiate metabolic acidosis in the affected tissue. We tested the hypothesis that acidotoxicity and acid-sensing ion channels (ASICs) are involved in the pathophysiology of ALS. We found that motoneurons were selectively vulnerable to acidotoxicity in vitro, and that acidotoxicity was partially reduced in asic1a-deficient motoneuron cultures. Cross-breeding of SOD1^G93A ALS mice with asic1a-deficient mice delayed the onset and progression of motor dysfunction in SOD1 mice. Interestingly, we also noted a strong increase in ASIC2 expression in motoneurons of SOD1 mice and sporadic ALS patients during disease progression. Pharmacological pan-inhibition of ASIC channels with the lipophilic amiloride derivative, 5-(N,N-dimethyl)-amiloride hydrochloride, potently protected cultured motoneurons against acidotoxicity, and, given post-symptom onset, significantly improved lifespan, motor performance and motoneuron survival in SOD1 mice. Together, our data provide strong evidence for the involvement of acidotoxicity and ASIC channels in motoneuron degeneration, and highlight the potential of ASIC inhibitors as a new treatment approach for ALS.     

Isolation and Structure of Ascochlorin and Its Analogs

Ascochlorin and its analogs were isolated from the filter cake of the fermented broth of a fungus, Ascochyta viciae Libert; the compounds obtained were ascochlorin, LL–Z1272 δ, LL–Z1272 ε, ascofuranone, ascofuranol and a new analog, C₂₃H₃₁ClO₅. Some of these prenyl phenols show hypolipidemic activity in both normolipidemic and hyperlipidemic rats. Details of the structure determination for ascochlorin are presented. The structure of the new analog was elucidated as 4′-hydroxy-5′-hydroascochIorin.     

Molecular dynamics of a far positioned SOD1 mutant V14M reveals pathogenic misfolding behavior

Human superoxide dismutase (Cu/Zn SOD1) is a homodimeric enzyme. Mutations in Cu/Zn SOD1 causes a familial form of amyotrophic lateral sclerosis (fALS), and aggregation of mutant SOD1 has been proposed to play a role in neurodegeneration. Though a majority of the mutations are point substitutions, there are a few changes that result in amino acid deletions or truncations of the polypeptide. These pathogenic mutations are scattered throughout the three-dimensional structure of the dimeric enzyme, which creates a puzzling pattern to investigate the molecular determinants of fALS. The most common hypothesis proposed that the misfolding of SOD1 mutants are primarily triggered by decreased affinity for metal ions. However, this hypothesis is challenging, as a significant number of disease-causing mutations are located far away from the metal-binding site and dimer interface. So in the present study, we have investigated the influence of such a far positioned pathogenic mutation, V14M, in altering the stability and folding of the Cu/Zn SOD1. Though the location of Val14 is far positioned, it has a vital role in the stability of SOD1 by preserving its hydrophobic cluster at one end of the β barrel domain. We have performed MD simulations of the V14M mutant for 80 ns timescale. The results reveal the fact that irrespective of its location, V14M mutation triggers a conformational change that is more similar to that of the metal-deficient holo form and could resemble an intermediate state in the folding reaction which results in protein misfolding and aggregation.     

Structure,gene expression,and evolution of primate copper chaperone for superoxide dismutase

Copper chaperone for superoxide dismutase (CCS) is essential for transporting copper ion to Cu,Zn-superoxide dismutase (Cu,Zn-SOD). We cloned cDNAs for six primate species' CCSs. The total number of amino acid residues of primate CCSs is 274. Similarities between primates were over 96%. Important residues for the CCS function were well conserved. A phylogenetic tree of CCSs and Cu,Zn-SODs from various organisms showed that these two proteins were derived from a common ancestor, diverging very early on during eukaryote evolution. The high frequency of nonsynonymous substitutions was found in the lineage to Old World monkeys and apes. Expression of the CCS gene in various tissues of Japanese monkey was found to be high in the liver and adrenal gland, followed by the kidney and small intestine. Such expressional pattern was similar with that of Cu,Zn-SOD gene (Fukuhara et al., 2002).     

阔叶红松混交林林隙大小和掘根微立地对小气候的影响

在小兴安岭阔叶红松混交林2.55 hm²样地内,选取由掘根倒木形成且具有坑和丘微立地的3个代表性林隙,并以空旷地和郁闭林分为对照,利用多通道HOBO自动气象站于2011年7-9月测定了不同大小林隙中心和丘顶部的光合有效辐射(PAR)、气温、相对湿度以及林隙中心的总辐射和降水量,比较了不同月份不同大小林隙中心和丘顶部微气候因子的差异,分析了不同大小林隙中心微气候因子的月变化以及不同大小林隙在典型天气条件下林隙中心和丘顶部微气候因子的日变化.结果表明: 3个不同大小林隙的月均PAR和月均气温排序是大林隙>中林隙>小林隙,月均相对湿度排序是小林隙>中林隙>大林隙;同一林隙中,丘顶部月均PAR和月均气温大于林隙中心,月均相对湿度为林隙中心>丘顶部;不同大小林隙和对照月均总辐射与月均气温均为7月>8月>9月,空旷地>大林隙>中林隙>小林隙>郁闭林分,月均相对湿度为郁闭林分>小林隙>中林隙>大林隙>空旷地.郁闭林分与各林隙以及与空旷地之间的月均相对湿度差异均显著;7-9月总降水量按照空旷地、大林隙、中林隙、小林隙、郁闭林分的次序依次递减;无论晴天与阴天,丘顶部日均PAR和日均气温都大于林隙中心,日均相对湿度则相反;无论丘顶部还是林隙中心,晴天日均PAR和日均气温都大于阴天,日均相对湿度则为阴天>晴天.