晶体氨基酸替代鱼粉蛋白对半滑舌鳎稚鱼消化酶和代谢酶活力的影响

为研究人工微颗粒饲料中晶体氨基酸替代鱼粉蛋白对半滑舌鳎（Cynoglossus semilaevis Gnther）稚鱼消化酶和代谢酶活力的影响, 以晶体氨基酸混合物分别替代0%、25%、50%、75%和100%鱼粉蛋白（0%CAA、25%CAA、50%CAA、75%CAA和100%CAA）, 在25%替代水平设计棕榈酸甘油酯包被晶体氨基酸混合物组（C-25%CAA）, 配制实验微颗粒饲料。每种微颗粒饲料随机投喂三组实验鱼初始体重（0.0940.02） g, 35日龄, 每组实验鱼放养150尾, 养殖周期28d。研究结果表明, 在不包膜条件下, 各处理组的胰蛋白酶活力随替代水平的升高显著下降（P0.05）, 且包膜处理组（C-25%CAA）与全鱼粉组无显著差异（P0.05）。肠段胰蛋白酶活力与胰段胰蛋白酶活力比值随氨基酸替代水平的升高显著下降（P0.05）, 鱼粉组显著高于75%和100%处理组（P0.05）, 但与25%处理组、包膜处理组（C-25%CAA）和50%处理组无显著差异（P0.05）。各处理组淀粉酶活力随替代水平的升高显著上升（P0.05）。亮氨酸氨肽酶（LA）和碱性磷酸酶（AP）活力（肠段与刷状缘）均随替代水平的升高显著下降（P0.05）, 包膜处理组（C-25%CAA）与全鱼粉组无显著差异（P0.05）。谷丙转氨酶（GPT/ALT）和谷草转氨酶（GOT/AST）活力随替代水平的升高显著上升（P0.05）, 50%、75%和100%处理组显著高于鱼粉组、25%处理组和包膜处理组（C-25%CAA）。研究结果显示, 饲料中晶体氨基酸显著影响了半滑舌鳎稚鱼的消化酶和代谢酶活力, 而且在25%的替代水平下, 与未包膜组相比, 包膜处理组能显著促进半滑舌鳎稚鱼消化系统的发育。       

2010年CLSI头孢菌素折点改变对产ESBLs奇异变形杆菌药物敏感性结果评估和分析

目的评估2009年CLSI M100-S19及2010年CLSI M100-S20文件中头孢他啶（CAZ）、头孢噻肟（CTX）和头孢唑啉（CFZ）最低抑菌浓度新旧折点变化对产ESBLs奇异变形杆菌药物敏感性试验结果的影响。方法对临床分离33株奇异变形杆菌进行产ESBLs菌株的确证试验,琼脂稀释法检测最低抑菌浓度（MIC）,根据药物敏感性结果分别对产ESBLs奇异变形杆菌和非产ESBLs奇异变形杆菌在S19和S20新旧折点下CAZ、CTX和CFZ三种药物的敏感性以及ESBLs阳性菌株分布率进行界定。结果 33株奇异变形杆菌中,产ESBLs菌株6株（18.2%）,由旧折点下耐药率50%（CAZ）、50%（CTX）和66.7%（CFZ）分别上升为新折点下66.7%（CAZ）、100%（CTX）和100%（CFZ）,敏感率由旧折点下33.3%（CAZ）和50%（CTX）分别下降为新折点下16.7%（CAZ）和0%（CTX）,CFZ在新旧折点下均为0,新旧折点标准下药敏结果分布率的差异均有统计学意义。结论对CTX、CFZ,按照CLSI S20新折点琼脂稀释法MICs折点标准判读药敏结果与ESBLs表型检测结果具有高度一致性,临床可以根据药敏结果选择用药,无需进行产ESBLs检测。但对于CAZ,尚需要进一步深入研究和评价。     

黄体酮对3T3-L1脂肪前体细胞成脂分化后细胞中UCP2基因表达的影响

目的：观察体外培养条件下3T3-L1脂肪前体细胞诱导分化成的成熟脂肪细胞中解偶联蛋白2（UCP2）mRNA表达水平及黄体酮对其表达的影响。方法：体外培养3T3-L1脂肪细胞,在诱导3T3．L1脂肪细胞分化成熟后,经不同黄体酮浓度10μm／25μM／50μM／75μM/100μM刺激后,抽提总RNA,用RT—PCR检测UCP2mRNA的表达。结果：黄体酮会促进成熟脂肪细胞中UCP2mRNA的表达,（P〈0．05）其中25μM浓度刺激下UCP2mRNA表达量最高。结论：体外培养中,黄体酮对成熟脂肪细胞中UCP2mRNA的表达与调控具有一定的影响。     

俄罗斯大果沙棘（Hippophae rhamnoides L.）种子萌发特性

俄罗斯丘依斯克大果沙棘（Hippophae rhamnoides L.）为优良的沙棘引进种。经测定,俄罗斯大果沙棘丘依斯克种子的千粒重为17.75g,与其它沙棘属的种子相比,其千粒重较大,为其2倍左右。四唑（TTC,1.0%）染色测种子的生活力的结果表明：有生活力的种子占97.75%,说明俄罗斯大果沙棘种子饱满度好;染色结果与对比发芽实验的结果很接近,说明用四唑染色来测定俄罗斯大果沙棘种子的发芽能力是较准确的方法。在水、气适宜的条件下,分别研究了温度分别为15、20、25、30℃,光照强度分别为25%、50%、75%、100%的不同处理对丘依斯克种子萌发的影响,并采用胚根生长量和全株生物量对沙棘种子发芽效果进行了评价,提出了沙棘种子发芽的最适温度和光强。结果表明,在25℃条件下,种子萌发最早,发芽势高达33.0%±4.76%,发芽率高达95.5%±1.5%,且主根生长量和生物量最大,分别为（4.5±0.09）cm和（0.137±0.002）g;75%光强最适合沙棘种子萌发,种子萌发最早,发芽势高达61.5%±1.7%,发芽率高达86.0%±1.1%,且主根生长量和生物量最大,分别为（3.7±0.2）cm和（0.108±0.004）g。因此,在直播营造大果沙棘林时,应首选25℃的温度条件,同时,建议进行适度遮荫处理。     

两步串联层析法纯化鼠抗人CD80单克隆抗体4E5

采用阴离子交换与凝胶过滤两步串联层析法,纯化了小鼠腹水来源的CD80阻断型单克隆抗体4E5。腹水样品经离心、过滤预处理后,在Tris-HCl缓冲溶液(pH8.0, 50mmol/L)条件下上阴离子交换柱对目的单抗进行捕集,采用0-0.5 mol/L NaCl浓度分步洗脱;含目的单抗的洗脱馏分再上凝胶过滤柱纯化,用PB缓冲溶液（pH7.2, 20mmol/L）洗脱,获得目的单抗4E5,其生物学活性高、纯度大于95%,抗体总回收率达61%。     

氮素形态配比对桔梗硝酸盐和亚硝酸盐动态积累及营养品质的影响

以桔梗（Platycodon grandiflorum）为试验材料,通过盆栽试验研究了等氮条件下6种氮素形态及铵硝氮配比(NH⁺₄ N/NO^-₃ N=100∶0、75∶25、50∶50、25∶75、0∶100、CO(NH₂)₂)对桔梗根中硝酸盐、亚硝酸盐动态积累以及营养、药用品质的影响。结果显示：（1）桔梗根中硝酸盐及亚硝酸盐积累量以铵硝比为25∶75处理下最低;硝酸盐积累量随栽培时间的增长呈上升趋势,尤其在10月采收时显著增加,亚硝酸盐变化趋势则与之相反。（2）桔梗根中Vc含量在全硝态氮处理下最高,可溶性多糖含量在铵硝比为50∶50处理下最高,而可溶性蛋白及总游离氨基酸含量均在铵硝比为75∶25处理下达到最大值。（3）桔梗根中N、Cu、Mn、Zn积累量在酰胺态氮处理下最高,其Fe、Mg、Cu积累量在铵硝比为75∶25处理下最大。（4）桔梗根中总黄酮含量随营养液中硝态氮比例增加而呈下降趋势,并在酰胺态氮处理下达到最大;桔梗多糖及桔梗总皂苷含量均在铵硝比为25∶75处理下有最大值。研究发现,在铵硝比为25∶75处理下,桔梗根中硝酸盐及亚硝酸盐含量最低,桔梗多糖及总皂苷积累量最高,且Vc、游离氨基酸等品质指标含量也较高,有利于桔梗品质的提升;由于10月采收时桔梗根中硝酸盐含量显著增高,桔梗采收前不宜大量追施氮肥。     

旱盐互作对冬小麦幼苗生长及其抗逆生理特性的影响

采用水培方法,以不同浓度的PEG-6000（0、8.3%、12.6%（W/V））和NaCl（0、25、50 mmol·L^-1）溶液模拟不同程度的干旱胁迫及盐胁迫,研究了盐分对干旱胁迫条件下冬小麦沧-6001幼苗生长及其抗逆生理特性的影响.结果表明：在8.3%或12.6% PEG-6000处理条件下,添加25 mmol·L^-1NaCl均使植株干物质积累和植株含水量比单一PEG处理增加,同时叶片可溶性糖和可溶性蛋白质含量增加,丙二醛和脯氨酸含量下降,植株各部位Na⁺含量升高、K⁺含量下降;在12.6% PEG-6000处理条件下,添加50 mmol·L^-1NaCl对植株的胁迫效应高于单一PEG处理.表明在干旱胁迫条件下,加入适量盐分可缓解干旱胁迫对冬小麦幼苗生长的抑制.     

氮素形态及配比对毛竹和青冈实生苗生长特性的影响

为探明毛竹及青冈对铵态氮和硝态氮的偏好吸收特征,通过对毛竹及青冈实生苗的水培试验,设置5种铵硝比(100∶0、75∶25、50∶50、25∶75、0∶100)的营养液处理,测定上述两种植物的叶片SPAD值、根系发育参数、根系中硝酸还原酶活性(NRA)。毛竹实生苗在铵硝比75∶25处理下,其SPAD值、根长、根表面积、NRA显著高于其他处理(P0.05)。青冈实生苗在铵硝比25∶75处理下,其根长、根表面积显著高于其他处理(P 0.05); SPAD值在铵硝比0∶100处理下显著高于其他处理(P0.01)。青冈根系中NRA随营养液中硝态氮浓度增加呈线型上升趋势,但毛竹苗培养过程中则无此相关关系。综上,不同氮素形态及配比处理下,毛竹苗在铵态氮为主要氮源时生长优势更明显,青冈苗则对硝态氮同化利用能力更强。     

大型绿藻浒苔转化表达系统选择标记的筛选

主要研究了条浒苔对抗生素氯霉素和除草剂Basta的敏感性,以确定适合的阳性选择标记基因。应用不同浓度氯霉素(0、25、50、75、100、125μg/ml)和Basta(0、5、12.5、25、37.5、50μg/ml)对不同发育时期条浒苔细胞存活率影响进行了测定。实验结果表明:不同发育时期条浒苔对氯霉素和Basta的敏感性不同。其中最大浓度125μg/ml浓度的氯霉素在15d内对条浒苔孢子和小苗两个不同发育时期的细胞均难以达到全部杀死效果,相对存活率仍分别为1%和20%;而Basta对条浒苔孢子和小苗均具有很强的杀生作用,其中5μg/ml浓度的Basta在3d内可将条浒苔孢子全部杀死,12.5μg/ml浓度下约一周时间可以将浒苔小苗全部致死。本实验结果提示bar基因有可能成为浒苔基因工程较理想的选择标记基因。     

N-(1-萘乙酰基)-,N’-(4-氨替吡啉基)硫脲对小麦幼苗的几种生理效应

小麦（Triticumaestivum）品种温－2540的种子用0．1％的HgCl2灭菌10min，每份取300粒，分别用N－（1－萘乙酸基）－，N’－（4－氨替吡啉基）硫脲（NAT，先用少量二甲亚砜溶解后，再溶于蒸馏水中）和NAA各50、10、1．0、0．1mg·L－1及蒸馏水浸种17h后，播于铺有3层滤纸的培养皿中，加等量蒸馏水，于25℃培养箱中萌发。胚芽鞘伸长至一定长度后转移到培养缸的尼龙网上，于人工光照箱中进行水培（Hoagland完全营养液，25℃，每天光照11h，光照度1400lx）。苗展开叶片时，喷施与浸种相对应浓度的药液。种子萌发72h时，随机取30粒，用精…     

Nutritional value of waxy barleys for broiler chickens

Winter wheat was compared with a barley which has a normal amylopectin/amylose starch ratio, and waxy barleys which have essentially 100% amylopectin starch. In one trial, broiler chicks were fed on one of seven diets which had grain portions of all wheat (100W), 50% wheat–50% Compana (normal) barley (50W–50C), 25% wheat–75% Compana (25W–75C), 100% Compana (0W–100C), 50% wheat–50% Wapana (waxy) barley (50W–50Wa), 25% wheat–75% Wapana (25W–75Wa), or 100% Wapana (0W–100Wa). Body weights (g) at 7 weeks were (100W) 1659, (50W–50C) 1523, (25W–75C) 1401, (0W–100C) 1396, (50W–50Wa) 1503, (25W–75Wa) 1403 and (0W–100Wa) 1293. Dressing percentage was not affected by diet but mortality was greater on 0W–100C. Metabolizable energy (kJ/g dry matter) values determined on non-laying Leghorn hens were 12.93 ± 0.38 and 12.76 ± 0.46 for the Compana and Wapana barley, respectively. In a second trial, addition of a food-grade diastase at 2 g per kg of feed did not affect body weight on wheat-based diets, increased weights over corresponding control diets for Wapana-based diets but gave inconsistent results for Washunopana, a hull-less waxy barley with an active α-amylase-like activity on the starch granules. Group feed values indicated that enzyme supplements improved feed efficiency for barley diets but metabolizable energy values were similar for diets with or without diastase additions.     

Hepatic radiation injury in the rat.

The whole livers of rats were exposed intraoperatively to graded doses (0 to 75 Gy) of 137Cs gamma radiation. At various times (0 to 155 days) after liver irradiation, minimally invasive, nondestructive tests (rose bengal retention and plasma alkaline phosphatase, glutamic-oxaloacetic acid transaminase, glutamic-pyruvic transaminase) were performed on at least half the surviving animals in each dose group to assess developing liver injury. Liver histology was done on animals sacrificed 96 to 100 days after irradiation. Radiation damage to the stomach killed approximately 50% of the animals 30 to 60 days after exposure to doses of 25 Gy or higher. These deaths were significantly reduced when care was taken to shield the stomach during irradiation. Stomach injury did not, however, appreciably affect liver function as measured by rose bengal retention. Whole-liver irradiation to 15 Gy resulted in reduced liver size and minimal histological changes, but did not result in increased rose bengal retention or plasma alkaline phosphatase concentration. The next highest dose group studied (25 Gy) showed significant histological abnormalities and liver injury as measured by increased rose bengal retention and liver enzymes. The latent period for development of hepatic injury, as measured by increased rose bengal retention, was 35 to 42 days and was relatively invariant over the 25- to 75-Gy dose range. Hepatic vein lesions and cellular necrosis were the most prominent histological lesions observed in 25-Gy-irradiated liver.     

The effect of seminal plasma on alpaca sperm function

In order to advance the development of assisted reproductive technologies in alpacas and other Camelids, the objective of this study was to explore the role of seminal plasma concentration on motility and functional integrity of alpaca sperm. Sixteen male alpacas > 3 y of age were used. In Experiment 1, epididymal sperm were incubated for 0 to 6 h in 0, 10, 25, 50, or 100% seminal plasma and motility was assessed. In Experiment 2, epididymal sperm were incubated in 0, 10, or 100% seminal plasma for 3 h and motility, acrosome integrity and DNA integrity were assessed. In Experiment 3, ejaculated sperm were incubated in 10, 25, 50, or 100% seminal plasma for 0 to 6 h and motility assessed. In Experiment 4, ejaculated sperm were incubated in 10 or 100% seminal plasma for 3 h and motility, acrosome integrity, DNA integrity, and viability were assessed. Epididymal and ejaculated sperm maintained motility longer when incubated in the presence of 10% seminal plasma compared to 0, 25, 50, or 100% seminal plasma (P < 0.001). The mean ± SEM percentage of epididymal sperm with intact acrosomes was less (P < 0.001) in samples incubated in 0% seminal plasma (39.4 ± 3.73) compared to 10% (75.3 ± 1.20) or 100% (77.4 ± 0.90) within 1 h after incubation. However, DNA integrity of ejaculated and epididymal sperm was not significantly affected by seminal plasma concentration. The mean viability of ejaculated sperm was reduced in the presence of 100 (12.7 ± 2.33) compared to 10% (36.2 ± 4.68) seminal plasma (P < 0.001) within 1 h of incubation. We concluded that alpaca semen should be diluted to a final concentration of 10% seminal plasma to prolong motility, preserve acrosome integrity, and maintain viability of sperm.     

Automated determination of tramadol enantiomers in human plasma using solid-phase extraction in combination with chiral liquid chromatography

A sensitive and automated method for the separation and individual determination of tramadol enantiomers in plasma has been developed using solid-phase extraction (SPE) on disposable extraction cartridges (DECs) in combination with chiral liquid chromatography (LC). The SPE operations were performed automatically by means of a sample processor equipped with a robotic arm (ASPEC system). The DEC filled with ethyl silica (50 mg) was first conditioned with methanol and phosphate buffer, pH 7.4 A 1.0-ml volume of plasma was then applied on the DEC. The washing step was performed with the same buffer. The analytes were eluted with 0.15 ml of methanol, and 0.35 ml of phosphate buffer, pH 6.0, containing sodium perchlorate (0.2 M) were added to the extract before injection into the LC system. The enantiomeric separation of tramadol was achieved using a Chiralcel OD-R column containing cellulose tris-(3,5-dimethylphenylcarbamate) as chiral stationary phase. The mobile phase was a mixture of phosphate buffer, pH 6.0, containing sodium perchlorate (0.2 M) and acetonitrile (75:25). The mobile-phase pH and the NaClO₄ concentration were optimized with respect to enantiomeric resolution. The method developed was validated. Recoveries for both enantiomers of tramadol were about 100%. The method was found to be linear in the 2.5–150 ng/ml concentration range [r²=0.999 for (+)- and (−)-tramadol]. The repeatability and intermediate precision at a concentration of 50 ng/ml were 6.5 and 8.7% for (+)-tramadol and 6.1 and 7.6% for (−)-tramadol, respectively.     

Determination of cimetidine in human plasma by high-performance liquid chromatography following liquid-liquid extraction

A new method is described for the determination of cimetidine in human plasma. The drug and internal standard (ranitidine) were separated on a Nucleosil C₁₈ 5 μm (25 × 4.6 mm I.D.) column using a mobile phase of acetonitrile-phosphate buffer, pH 6.2 (25:75, v/v) containing 2.5 g/l heptane sulphonic acid. The mobile phase was delivered at a flow-rate of 0.9 ml/min, detection was by ultraviolet absorption at 228 nm and concentrations were calculated on the basis of peak areas. The drugs were extracted from alkaline plasma into ethyl acetate using a salting out procedure which involved the addition of 100 ml of a saturated solution of K₂CO₃ to each 250-μl plasma aliquot. The method was validated over the concentration ranges 50–3000 ng/ml and 100–7000 ng/ml for two separate studies. Mean coefficients of variation were less than 6% for both intra- and inter-assay in both studies and recoveries varied between 71 and 81%. The method was successfully applied to the determination of cimetidine in plasma for a pharmacokinetic study.     

Effect of carbamazepine and gabapentin on excitability in the trigeminal subnucleus caudalis of neonatal rats using a voltage-sensitive dye imaging technique

Background

The antiepileptic drugs carbamazepine and gabapentin are effective in treating neuropathic pain and trigeminal neuralgia. In the present study, to analyze the effects of carbamazepine and gabapentin on neuronal excitation in the spinal trigeminal subnucleus caudalis (Sp5c) in the medulla oblongata, we recorded temporal changes in nociceptive afferent activity in the Sp5c of trigeminal nerve-attached brainstem slices of neonatal rats using a voltage-sensitive dye imaging technique.

Results

Electrical stimulation of the trigeminal nerve rootlet evoked changes in the fluorescence intensity of dye in the Sp5c. The optical signals were composed of two phases, a fast component with a sharp peak followed by a long-lasting component with a period of more than 500 ms. This evoked excitation was not influenced by administration of carbamazepine (10, 100 and 1,000 μM) or gabapentin (1 and 10 μM), but was increased by administration of 100 μM gabapentin. This evoked excitation was increased further in low Mg²⁺ (0.8 mM) conditions, and this effect of low Mg²⁺ concentration was antagonized by 30 μM DL-2-amino-5-phosphonopentanoic acid (AP5), a N-methyl-d-aspartate (NMDA) receptor blocker. The increased excitation in low Mg²⁺ conditions was also antagonized by carbamazepine (1,000 μM) and gabapentin (100 μM).

Conclusion

Carbamazepine and gabapentin did not decrease electrically evoked excitation in the Sp5c in control conditions. Further excitation in low Mg²⁺ conditions was antagonized by the NMDA receptor blocker AP5. Carbamazepine and gabapentin had similar effects to AP5 on evoked excitation in the Sp5c in low Mg²⁺ conditions. Thus, we concluded that carbamazepine and gabapentin may act by blocking NMDA receptors in the Sp5c, which contributes to its anti-hypersensitivity in neuropathic pain.     

卡马西平降解菌的筛选及降解特性研究

药品和个人护理品类污染物日益成为新兴污染物研究的重点, 药品卡马西平因具有多种药效被广泛使用, 在环境中频繁被检出, 且浓度较高, 不易去除, 通常作为环境中药品和个人护理品污染状况的指示化合物。本研究从某制药厂的污水处理厂中分离到一株细菌HY-7, 能以卡马西平为唯一碳源、氮源和能源生长, 通过生理生化以及16S rDNA、gyrB基因序列分析鉴定并命名为Acinetobacter sp. HY-7。该菌株生长和降解卡马西平的最适条件为25°C和pH 6.0, 经HPLC分析10 d内能将初始浓度为20 mg/L的卡马西平降解48%。菌株HY-7还能以邻苯二酚、吲哚、萘、蒽等芳香族化合物为唯一碳源生长。     

Selective inhibition of brain Na,K-ATPase by drugs

The effect of drugs from the class of cardiac (methyldigoxin, verapamil, propranolol), antiepileptic (carbamazepine), sedative (diazepam) and antihistaminic (promethazine) drugs on Na,K-ATPase activity of plasma membranes was studied in rat brain synaptosomes. Methyldigoxin in a concentration of 0.1 mmol/l inhibits enzyme activity by 80 %. Verapamil, propranolol and promethazine in concentrations of 20, 20 and 2 mmol/l respectively, entirely inhibit the ATPase activity. Carbamazepine and diazepam in concentrations of 0.02-60 mmol/l have no effect on the activity of this enzyme. According to the drug concentrations that inhibit 50 % of enzyme activity (IC(50)), the potency can be listed in the following order: methyldigoxin promethazine verapamil ? propranolol. From the inhibition of commercially available purified Na,K-ATPase isolated from porcine cerebral cortex in the presence of chosen drugs, as well as from kinetic studies on synaptosomal plasma membranes, it may be concluded that the drugs inhibit enzyme activity, partly by acting directly on the enzyme proteins. Propranolol, verapamil and promethazine inhibitions acted in an uncompetitive manner. The results suggest that these three drugs may contribute to neurological dysfunctions and indicate the necessity to take into consideration the side effects of the investigated drugs during the treatment of various pathological conditions.     

The anaerobic degradability of thermoplastic starch: polyvinyl alcohol blends: potential biodegradable food packaging materials

A systematic study on the anaerobic degradability of a series of starch:polyvinyl alcohol (TPS:PVOH) blends was performed to determine their fate upon disposal in either anaerobic digesters or bioreactor landfills. The aims of the study were to measure the rate and extent of solubilisation of the plastics. The extent of substrate solubilisation on a COD basis reached 60% for a 90:10 (w/w) blend of TPS:PVOH, 40% for 75:25, 30% for 50:50 and 15% for PVOH only. The rate of substrate solubilisation was most rapid for the 90:10 blend (0.041 h(-1)) and decreased with the amount of starch in the blend in the following order 0.034 h(-1)(75:25); 0.023 h(-1)(50:50). The total solids that remained after 900 h were 10 wt.% (90:10); 23 wt.% (75:25); 55 wt.% (50:50); 90 wt.% (0:100). Starch containing substrates produced a higher concentration of volatile fatty acids (VFAs) and biogas, compared to the 0:100 substrate. The major outcome was that PVOH inhibited the degradation of the starch from the blend.