犬恶丝虫核苷二磷酸激酶(NDPK)基因的原核表达及其免疫荧光定位

对犬恶丝虫核苷二磷酸激酶(NDPK)基因的基本特征进行探究,并评估其诊断价值,旨在为犬恶丝虫病的诊断提供依据。本研究原核表达了Di-NDPK,通过生物信息学、免疫印迹和免疫荧光组化分析了该蛋白的基本特征,通过检测犬恶丝虫阳性血清评估了rDi-NDPK的诊断价值。免疫印迹显示,Di-NDPK具有良好的反应原性,免疫荧光组化显示,Di-NDPK在犬恶丝虫的侧索及肠上皮细胞和肠腔中分布。间接ELISA显示,该方法的敏感性为66.7%(16/24),特异性为38.9%(14/36),其中与犬细粒棘球蚴病阳性血清、犬钩虫病阳性血清以及犬弓首蛔虫病阳性血清发生交叉反应。Di-NDPK是一个分泌蛋白,但不适合作为犬恶丝虫病的诊断候选抗原。     

Nucleoside by‐product dietary supplementation influences blood chemistry,immune response,oxidative stress resistance and intestinal morphology of juvenile amberjack,Seriola dumerili

We explored the influence of nucleoside by‐products (NBP) on blood chemistry, immune response, oxidative stress and intestinal morphology of Seriola dumerili. Four experimental diets were formulated where diets 1–3 are semipurified and supplemented with liquid NBP at concentration of 0 (D1, negative control), 30 (D2) and 90 (D3) g/kg. Diet 4 (D4) is a fishmeal‐based positive control diet. Each diet was randomly allocated to triplicate groups of fish for 50 days. The results revealed a significant influence of NBP supplementations on total cholesterol, blood urea nitrogen and triglyceride level. Lysozyme activity (LA), total serum protein (TSP) and peroxidase activity (PA) also increased with NBP supplementation as compared to the negative control. TSP, PA and LA were numerically higher in diet groups D2 and D4, respectively. Anterior enterocyte height (hE) was higher in diet group D4, while NBP‐supplemented groups showed intermediate values with D1 group and negative control group showed significantly lowest value. Supplementation of NBP also increased anterior fold height (hF) and microvillus height and posterior hF, hE and microvillus height compared with D1. Finally, we concluded that NBP can be used as a cheaper nucleotide/nucleoside source for amberjack. Supplementation of more than 30 g/kg in diets improved blood chemistry, non‐specific immunity, oxidative stress resistance and intestinal morphology of amberjack.     

嘌呤核苷磷酸化酶温敏型表达载体的构建、表达及应用于腺苷转化的研究

通过双酶切方法将重组质粒上大肠杆菌嘌呤核苷磷酸化酶(PNPase)基因克隆到pBV220表达载体上,构建了温敏型PNPase的表达载体pBV 220-PNP,转化大肠杆菌DH5α并使之表达.SDS-PAGE电泳结果显示出明显的特异表达条带,其分子量约为26 kDa.在此基础上研究了不同诱导条件对蛋白表达量的影响.分别以肌苷和尿苷为核糖供体研究重组菌转化腺苷的条件,发现肌苷最适合作为核糖供体,在30 mmol.L-1pH 7.5的Na2HPO4-KH2PO4缓冲液中,以30 mmol.L-1肌苷和10 mmol.L-1腺嘌呤作底物,加入0.5%基因工程湿菌体60℃反应3 h,腺苷的转化率为85.57%.     

周期性“饥饿-再投喂”对大口黑鲈幼鱼补偿生长的影响

在水温(26.5±1.2)℃下,将体质量(45.70±0.56)g的大口黑鲈Micropterus salmoides幼鱼分为4组,饲养在采光大棚内的30m2水泥池中,每池40尾,D0组每天饱食投喂2次,D1、D2、D3组分别投喂6d、5d、4d,饥饿1d、2d、3d,持续8周,每组3个重复池塘,研究周期性“饥饿-再投喂”对大口黑鲈幼鱼补偿生长的影响。结果发现,随着每周饥饿天数的增加,大口黑鲈的末体质量和增重率均不同程度下降,但D1组与D0组无显著差异(P>0.05)。大口黑鲈的日摄食量随饥饿时间的增加而显著提高(P<0.05),但各组间无显著差异(P>0.05)。D1组的饲料利用效率和蛋白质效率与D0组无显著差异(P<0.05);而D2和D3组显著低于D0组(P<0.05)。试验结束时,鱼体粗蛋白和粗脂肪含量随饥饿时间增加而下降。D3组鱼的胃和肠道蛋白酶活性显著低于D0组(P<0.05);D1组的胃和肠道脂肪酶活性较高;D1和D2组胃淀粉酶活性较高,肠道淀粉酶活性则随着饥饿时间增加显著上升(P<0.05)。饥饿使大口黑鲈血清甘油三酯和总胆固醇含量降低,血清谷草转氨酶和谷丙转氨酶活性增加。血清生长激素水平随着饥饿时间增加而上升,类胰岛素生长因子-Ⅰ水平则下降(P<0.05)。结果表明:每周投喂6d饥饿1d的大口黑鲈饲料转化效率较高,实现了完全补偿生长,可供集约化养殖的科学投喂参考。     

灵芝有效化学成分的研究

本文从灵芝有效化学成分的结构，提取分离方法，结构鉴定，生理活性及应用价值等方面进行了概述。     

An in vitro biomechanical comparison of a limited-contact dynamic compression plate fixation with a dynamic compression plate fixation of osteotomized equine third metacarpal bones

OBJECTIVES: To compare the monotonic biomechanical properties and fatigue life of a broad, limited contact, dynamic compression plate (LC-DCP) fixation with a broad, dynamic compression plate (DCP) fixation to repair osteotomized equine 3rd metacarpal (MC3) bones. STUDY DESIGN: In vitro biomechanical testing of paired cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by 1 of 2 methods for fracture fixation. ANIMAL POPULATION: Twelve pairs of adult equine cadaveric MC3 bones. METHODS: Twelve pairs of equine MC3 were divided into 3 test groups (4 pairs each) for (1) 4-point bending single cycle to failure testing, (2) 4-point bending cyclic fatigue testing, and (3) torsional single cycle to failure testing. An LC-DCP (8-hole, 4.5 mm) was applied to the dorsal surface of 1 randomly selected bone from each pair. One DCP (8-hole, 4.5 mm broad) was applied dorsally to the contralateral bone from each pair. All plates and screws were applied using standard AO/ASIF techniques to MC3 bones that had mid-diaphyseal osteotomies. Mean test variable values for each method were compared using a paired t-test within each group. Significance was set at P<.05. RESULTS: The mean 4-point bending yield load, yield bending moment, composite rigidity, failure load, and failure bending moment of LC-DCP fixation were significantly greater (P<.01) than those of broad DCP fixation. Mean cycles to failure for 4-point bending was significantly (P<.001) greater for broad DCP fixation compared with broad LC-DCP fixation. Mean yield load, mean composite rigidity, and mean failure load in torsion was significantly (P<.02) greater for broad LC-DCP fixation compared with broad DCP fixation. CONCLUSION: Broad LC-DCP offers increased stability in static overload testing, however, it offers significantly less stability in cyclic fatigue testing. CLINICAL RELEVANCE: The clinical relevance of the cyclic fatigue data supports the conclusion that the broad DCP fixation is biomechanically superior to the broad LC-DCP fixation in osteotomized equine MC3 bones despite the results of the static overload testing.     

An in vitro evaluation of plate luting using osteotomized equine third metacarpal bones with a limited contact-dynamic compression plate

OBJECTIVES: To evaluate the effects of plate luting on the biomechanical properties of a broad limited contact-dynamic compression plate (LC-DCP) fixation to repair osteotomized equine 3rd metacarpal (MC3) bones. STUDY DESIGN: In vitro biomechanical testing of paired cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by LC-DCP fixation, with 1 of the pair luted with polymethylmethacrylate (PMMA). ANIMAL POPULATION: Ten pairs of adult equine cadaveric MC3 bones. METHODS: Ten pairs of equine MC3 were divided into 2 test groups (5 pairs each) for (1) palmarodorsal 4-point bending single cycle to failure testing and (2) palmarodorsal 4-point bending cyclic fatigue testing. The LC-DCP (8 hole, 4.5 mm) was applied to the dorsal surface of each pair of MC3 bones. All plates and screws were applied using standard AO/ASIF techniques. All MC3 bones had mid-diaphyseal osteotomies. One of the matched pairs of LC-DCP-MC3 constructs were randomly chosen to be luted with PMMA. Mean test variable values for each method were compared using a paired t-test within each group; significance was set at P<.05. RESULTS: Mean palmarodorsal 4-point bending yield bending moment, failure bending moment of the LC-DCP fixation with luting was not significantly different (P>.05) than those of the LC-DCP fixation without luting. Mean cycles to failure for palmarodorsal 4-point bending was significantly (P<.0003) greater, with a 7.2-fold increase, for the LC-DCP fixation with luting compared with the LC-DCP fixation without luting. CONCLUSION: Luting the broad LC-DCP with PMMA in the fixation osteotomized equine MC3 bones increases the fatigue life of cyclic loading for palmarodorsal 4-point bending under the in vitro conditions studied. CLINICAL RELEVANCE: The cyclic fatigue data supports the conclusion that luted broad LC-DCP fixation is biomechanically superior to the non-luted broad LC-DCP fixation in osteotomized equine MC3 bones.     

An in vitro biomechanical comparison of a prototype equine metacarpal dynamic compression plate fixation with double dynamic compression plate fixation of osteotomized equine third metacarpal bones

OBJECTIVES: To compare the monotonic biomechanical properties of a prototype equine third metacarpal dynamic compression plate (EM-DCP) fixation with a double broad dynamic compression plate (DCP) fixation to repair osteotomized equine third metacarpal (MC3) bones. STUDY DESIGN: In vitro biomechanical testing of paired cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by 1 of 2 methods for fracture fixation. POPULATION: Twelve pairs of adult equine cadaveric MC3 bones. METHODS: Twelve pairs of equine MC3 were divided into 3 test groups (4 pairs each) for (1) 4-point bending single cycle to failure testing, (2) 4-point bending cyclic fatigue testing, and (3) torsional testing. The EM-DCP (10-hole, 4.5 mm) was applied to the dorsal surface of one randomly selected bone from each pair. Two DCPs, 1 dorsally (10-hole, 4.5 mm broad) and 1 laterally (9-hole, 4.5 mm broad) were applied to the contralateral bone from each pair. All plates and screws were applied using standard AO/ASIF techniques to MC3 bones that had mid-diaphyseal osteotomies. Mean test variable values for each method were compared using a paired t-test within each group. Significance was set at P<.05. RESULTS: Mean 4-point bending yield load, yield bending moment, bending composite rigidity, failure load and failure bending moment of the EM-DCP fixation were significantly greater (P<.0001) than those of the double broad DCP fixation. Mean cycles to failure in 4-point bending of the EM-DCP fixation was significantly greater (P<.0008) than that of the double broad DCP fixation. Mean yield load, composite rigidity, and failure load in torsion of the EM-DCP fixation were significantly greater (P<.0035) than that of the double broad DCP fixation. CONCLUSION: The EM-DCP provides increased stability in both static overload testing and cyclic fatigue testing. CLINICAL RELEVANCE: Results of this in vitro study support the conclusion that the prototype EM-DCP fixation is biomechanically superior to the double broad DCP fixation for the stabilization of osteotomized equine MC3.     

Differential aphicidal effects of chitinase inhibitors on the polyphagous homopteran Myzus persicae (Sulzer)

Four chitinase inhibitors, cyclo-(Proline-Tyrosine), cyclo-(Histidine-Proline), allosamidin and psammaplin A, were selected for in vitro feeding experiments with the peach-potato aphid, Myzus persicae (Sulzer), under controlled photoperiod and temperature conditions. Artificial diets were used to provide chitinase inhibitors at 10, 50 and 100 microg mL(-1) to M. persicae. Except for cyclo-(Proline-Tyrosine), which did not modify aphid demographic parameters, chitinase inhibitors induced differential aphicidal effects on M. persicae. At all doses, cyclo-(Histidine-Proline) induced significant effects affecting daily fecundity, intrinsic rate of natural increase (r(m)) and doubling time of population. When compared with the control diet, allosamidin decreased nymph survival and daily fecundity, increasing the doubling time of population from 1 to 1.5 days. Psammaplin A was the most toxic inhibitor when delivered via artificial diet, as it induced the death of all aphids reared at 50 and 100 microg mL(-1). The results demonstrate the potential use of chitinase inhibitors as aphid management tools.     

Development of LVL frame structures using glued metal plate joints II: strength properties and failure behavior under lateral loading

In past years high priority was given to developing a seismic design for wood structures, including research on the response of wood structures to earthquakes. In this study a new type of portal frame with relatively large span was developed for the traditional Japanese wooden houses with large openings at the front to strengthen the structure. Stainless steel plates coated with zinc and glued with epoxy adhesives on laminated veneer lumber (LVL) members, composed of Douglas fir veneer and bonded with phenolformaldehyde resin, were used. The connection between the frame's beam and columns and between the columns and groundsills was mechanical, with bolts. The subject of this research was to analyze strength properties and failure behavior of glued LVL metal joints used as structural components and to evaluate the response of LVL portal frames under cyclic lateral loading. The results show that portal frames using glued LVL metal plates have a good multiplier for the shear walls and may be applied to traditional Japanese structures. The equivalent viscous damping provided good energy dissipation in the frames. The joints displayed good mechanical behavior during tests; moreover, the structures demonstrated high strength, stiffness, and ductility, which are necessary for a seismic design.Part of this paper was presented at the 47th annual meeting of the Japan Wood Research Society, Kouchi, April 1997; and at the 5th world conference on timber engineering. Montreux, Switzerland, August 1998