枇杷果实发育过程中山梨醇含量及相关酶活性的变化

以果型和品质差异较大的黄肉枇杷(Eriobotrya japonica Lindl.)‘解放钟’和白肉枇杷‘白梨’的果实为试材,测定果实发育过程中山梨醇、果糖、葡萄糖含量及与山梨醇代谢相关酶--山梨醇脱氢酶(Sorbitol dehydrogenase,SDH)和山梨醇氧化酶(Sorbitol oxidase,SOX)活性的变化,并探讨山梨醇代谢与枇杷果实生长发育和品质形成的关系。结果表明:2个枇杷品种果实中山梨醇含量均随果实发育进程呈下降趋势,且‘白梨’枇杷果实中山梨醇含量高于‘解放钟’。在花后35~80 d,2个枇杷品种幼果中葡萄糖和果糖含量均较低且相对稳定,而花后95~140 d果实中葡萄糖和果糖含量逐渐上升,并在花后125~140 d果实成熟期急剧增加而达到峰值;果实中葡萄糖含量在2个品种间差异不显著(p0.05),花后125~140 d的果实成熟期,‘白梨’果实中果糖含量显著高于‘解放钟’(p0.05)。果实发育过程中SDH和SOX的活性均呈先降后升的变化趋势。说明2个枇杷品种果实发育过程中山梨醇、果糖、葡萄糖含量及SDH和山SOX的活性表现出相似的变化规律,但果实山梨醇含量与SDH和SOX的活性在果实膨大期均存在显著差异(p0.05),这可能是黄肉枇杷与白肉枇杷品质差异的关键。     

Effect of garlicin on viral myocarditis in mice

AIM: To investigate the effect of garlicin on mouse viral myocarditis and to explore the possible mechanisms. METHODS: Forty BALB/c mice were randomly divided into 4 groups: control group, viral myocarditis group, low-dose garlicin group and high-dose garlicin group. The morphological changes of the myocardium were observed with HE staining. The levels of lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and creatine kinase (CK) were measured by colorimetric method. The protein levels of Bcl-2, Bax, cleaved-caspase-9, cleaved-caspase-3, cleaved-PARP and cleaved-caspase-8 in the myocardial tissues were determined by Western blot. RESULTS: The morphological observation with HE staining showed that garlicin inhibited the necrosis of the myocardium and infiltration of inflammatory cells. Compared with viral myocarditis group, garlicin dose-dependently decreased the levels of LDL, AST and CK. Moreover, garlicin treatment decreased the protein levels of Bax, cleaved-caspase-9, cleaved-caspase-3 and cleaved-PARP, accompanied with an increase in Bcl-2 expression. However, garlicin had no effect on the protein level of cleaved-caspase-8. CONCLUSION: Garlicin effectively attenuates the myocardial damage in mice with viral myocarditis through inhibition of intrinsic apoptosis pathway.     

Assessment of Lactate Dehydrogenase,Alkaline Phosphatase and Aspartate Aminotransferase Activities in Cow’s Milk as an Indicator of Subclinical Mastitis

This study examined the activities of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in the milk of lactating Holstein cows in association with subclinical mastitis (SCM). A total of 94 milk samples were collected from 58 lactating dairy cows representing stages of lactation from the second to the tenth week after calving. Those which were classified as positive by California mastitis test (CMT) were deemed to have subclinical mastitis. All the milk samples were skimmed by centrifugation at 10 000g at 0 degrees C and were used for enzyme activities estimations. The mean activities of LDH and ALP were higher in the milk from udders with SCM than in the milk from healthy udders (p < 0.05). There were no significant differences in AST values. The maximum agreement rates between the CMT results and LDH and ALP values were seen at thresholds of > 180 IU/L and > 40 IU/L respectively (kappa values 0.65 and 0.79, respectively). However, the sensitivity of the tests for identifying SCM at these thresholds was higher for ALP (96.4%) than for LDH (68.5%). In this study, LDH and ALP tests were standardized for cow's milk and results showed that only the ALP test was reliable in the early diagnosis of subclinical mastitis.     

玉米大斑病菌甘油醛-3-磷酸脱氢酶基因克隆和表达分析

利用RT-PCR技术克隆玉米大斑病菌(Setosphaeria turcica)的GAPDH基因StGAPDH。该基因c DNA全长1 014 bp,编码337个氨基酸;蛋白质分子量36 505.44,理论pI值6.72,属弱酸、亲水、稳定蛋白,无跨膜结构域和信号肽。亚细胞定位于细胞质、线粒体和细胞核的概率分别为43.5%、30.4%和21.7%;氨基酸序列高度保守,在亲缘关系上与玉米小斑病菌(Bipolaris maydis)和水稻胡麻斑病菌(Bipolaris oryzae)最近。荧光定量PCR分析表明,在病菌接种抗病玉米自交系3 d时,StGAPDH基因表达量显著高于接种5～10 d(P<0.05);接种感病玉米自交系时,接种10 d的StGAPDH基因表达量显著高于接种3～7 d(P<0.05)。     

Molecular cloning,characterization and nutritional regulation of key enzymes required for the effective utilization of marine wax esters by Atlantic salmon (Salmo salar L.)

Previous studies had shown that wax ester‐rich lipid extracted from calanoid copepods could be a useful alternative to fish oil as a provider of long‐chain n‐3 polyunsaturated fatty acids in diets for use in salmon aquaculture. Effective utilization of wax ester requires digestion and metabolism in the intestine with the fatty alcohol component being oxidized to fatty acid in intestinal cells through the combined activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). We studied wax ester utilization in Atlantic salmon using a candidate gene approach, focusing on ADH and ALDH as sequence information was available for these genes, including fish sequences, facilitating isolation of the cDNAs. Here, we report on the isolation and cloning of full‐length cDNAs for ADH3 and ALDH3a2 genes from salmon intestinal tissue. Functional characterization by heterologous expression in the yeast, Saccharomyces cerevisiae, showed the products of these cDNAs had long‐chain ADH and ALDH enzyme activities. Thus, ADH3 was capable of oxidizing long‐chain fatty alcohol, and ALDH3a2 was capable of oxidizing long‐chain fatty aldehyde to the corresponding fatty acid. The genes were highly expressed in intestinal tissue, particularly pyloric caeca, but their expression was not increased in salmon fed dietary copepod oil in comparison to fish fed fish oil.     

纤维素酶催化水解和氧化机制的研究进展

人类对纤维素的开发与利用还非常有限,未能完全搞清天然纤维素的生物降解机制是其一个重要原因.自从C1-Cx假说提出以来,又有较多的降解理论出现对其改进,纤维素酶水解机制不能完全解释天然纤维素的降解,大量文献证实天然纤维素降解存在氧化机制.本文主要介绍水解与氧化机制中具代表性酶协同降解模型、纤维二糖水解酶(CBH)协同作用假说、羟基自由基的氧化机制和纤维二糖脱氢酶途径等不同降解理论,将水解和氧化两种机制结合起来给出其降解途径,也许能够较为容易理解纤维素的降解具体过程.     

Enzyme activity levels associated with the production of glycerol as an antifreeze in liver of rainbow smelt (Osmerus mordax)

Rainbow smelt (Osmerus mordax) tolerate temperatures close to the freezing point of sea water, in part, through the use of glycerol as an antifreeze. Potential mechanisms for glycerol production by liver were assessed by comparing activities of key enzym es of carbohydrate and amino acid metabolism in rainbow smelt to those in Atlantic tomcod (Microgadus tomcod) and smooth flounder (Liopsetta putmani). The latter two species inhabit the same environment but do not maintain high levels of blood glycerol. The enzyme profile of liver from rainbow smelt is substantially different from those of the other species and is poised for glycerol production. With respect to carbohydrate metabolism, glycerol-3-phosphate dehydrogenase activity in rainbow smelt liver was 156 µmoles min^-1 g^-1, a level which was 28 and 12-fold higher than activities in tomcod and flounder liver, respectively. Glycerol-3-phosphatase activity in smelt liver was 1.95 µmol min^-1 g^-1. This activity was 2.7 and 5.4-fold higher than those in tomcod and flounder liver, respectively. As such, the production of glycerol appears to be dependent upon the concerted action of glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase. The enzyme profile also suggests that amino acids are a potential source of carbon for glycerol. Aspartate aminotransferase activity in rainbow smelt was 7 to 14-fold higher in comparison to tomcod and flounder liver, respectively. Activities of alanine aminotransferase and glutamate dehydrogenase in liver were between 2 and 3-fold higher in rainbow smelt than in the other two species. Finally, it is shown that in vitro preparations of smelt liver sections produce glycerol at 0°C lending support to the concept that liver is a site of glycerol synthesis in vivo.     

2,3-丁二醇脱氢酶基因(BudC)过表达菌株的构建和初步鉴定

旨在过表达BudC进而上调2,3-丁二醇（2,3-Butanediol, 2,3-BD）的合成,通过基因工程手段构建整合载体pR1SW-BudC,经PCR和酶切双重验证后将其电转化至产酸克雷伯氏菌(Klebsiella oxytoca) HD79。结果经卡那霉素筛选,获得一株菌株K. oxytoca HD79-01。以原始菌株为对照,摇瓶发酵检测2,3-BD含量和相关酶活,q RT-PCR检测BudC表达差异,最终BudC在HD79-01中转录水平的表达量为HD79的45.25倍（p<0.01）。摇瓶发酵显示2,3-BD的最高产量、转化率和生产强度分别提高了24.54%、10.97%、45.08%（分别为30.818 ± 0.003 g/L、0.253 ± 0.013 g/g、0.43 ± 0.01 g/(L·h)）,酶活提高了3.61倍(0.563 ± 0.003 U/mg)。因此,2,3-丁二醇脱氢酶基因(BudC)过表达菌株构建不仅成功的提高了2,3-BD的产量也为实现2,3-BD的工业化生产奠定基础。     

黄瓜叶面喷施硅肥对果实糖分和维生素C积累及相关酶的影响

研究了黄瓜叶面喷施硅肥对其生长、产量、果实糖分、维生素C及相关酶活性和基因表达量的影响。结果表明：叶面喷施硅肥显著促进了黄瓜生长,增加了单株产量,提高了果实中碱性转化酶（NI）、蔗糖磷酸合成酶（SPS）、蔗糖合成酶（SS）及半乳糖内脂脱氢酶（GaILDH）活性,提高了果实果糖、葡萄糖和维生素C含量。其中,1.420 g · L^-1单硅酸处理效应最显著,其果糖、葡萄糖和维生素C含量分别较对照高13.79%、18.08%和75.11%。实时荧光定量PCR分析表明,叶面喷施单硅酸的黄瓜果实碱性转化酶–3（NI-3）、蔗糖磷酸合成酶（SPS）和半乳糖内脂脱氢酶（GaILDH）基因表达量显著提高。     

五汁饮对急性酒精中毒大鼠肝脏的保护机制研究

目的 研究五汁饮对急性酒精中毒大鼠肝脏的保护作用。方法 SD大鼠分为5组,分别为空白组、模型组、葛花组、五汁饮低剂量组及高剂量组,每组10只。预防性给药 7 d后,空白组蒸馏水灌胃,其余各组按照1.5 mL/100 g的52度红星二锅头给予灌胃,每日1次,连续1周,建立急性酒精性肝损伤模型。测定血清谷丙转氨酶（ALT）、谷草转氨酶（AST）含量;测定肝脏匀浆中谷胱甘肽（GSH）、超氧化物歧化酶（SOD）、丙二醛（MDA）、乙醇脱氢酶（ADH）含量。结果 与模型组相比,葛花组、五汁饮高剂量组ALT含量降低（P<0.05）,五汁饮高、低剂量组AST含量降低（P<0.01）;五汁饮高剂量组ADH含量高于模型组（P<0.05）,葛花组、五汁饮低剂量组与模型组没有统计学差异（P>0.05）;葛花组、五汁饮高、低剂量组GSH、SOD含量高于模型组（P<0.01,P<0.05）,葛花组、五汁饮高、低剂量组均可显著降低MDA含量（P<0.01）。结论 五汁饮保肝作用可能通过激活ADH加快酒精代谢及SOD、GSH的抗氧化作用来实现。