II型猪圆环病毒检测试剂盒的研制

猪圆环病毒(Porcine circovirus, PCV)属圆环病毒科圆环病毒属成员[1]。PCV分两种血清型,即PCV 1 和 PCV 2,其中 PCV 1 由 Tischer 等[2] 于1974年检测到,对猪没有致病性;PCV 2 可引起部分断奶仔猪和育肥猪的断奶仔猪多系统综合征( postweaning multisystemic wasting syndrome,PMWS)[3]。目前,该病呈世界分布,给养猪业造成巨大的经济损失。本研究通过分析已发表的 PCV 2的基因序列,找出 PCV 2 的特异性片段,研制了检测PCV 2的试剂盒。检测临床疑似病料,结果表明该试剂盒使用方便、快速、敏感、特异,符合国内的养殖场…     

Determination of the relative positions of amino acids by partial specific cleavages of end-labeled proteins

We have developed a new method for obtaining information about protein sequences that uses an approach analogous to that used to determine DNA sequences. In essence, three steps are involved. First, a detectable label is attached exclusively to the amino terminus of a polypeptide. Next, the labeled chain is subjected to partial specific cleavage in a way that produces roughly equimolar amounts of fragments of different sizes. Cleavages for methionine, tryptophan, arginine, aspartyl-proline bonds, and asparaginyl-glycine bonds have been employed. Lastly, the labeled fragments are separated according to size by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The distribution of target amino acids along the polypeptide chain can be deduced from the specific pattern of labeled bands by reading the "ladder" in the same way that DNA sequencing gels are read. Although the method can be conducted with a radioactive label, we have chosen to use a fluorescent label. We have applied the method successfully to the three subunit chains of two different fibrinogens.     

The polysaccharides from Ganoderma lucidum: Are they always inhibitors on human hepatocarcinoma cells?

The antitumor activity of intracellular polysaccharides from submerged fermentation of Ganoderma lucidum was investigated focusing on the inhibition on human liver cancer cells. The polysaccharides inhibited human hepatocarcinoma cell HepG2 during earlier phase with lower dosage but obviously became less functional in later phase regardless of the dosage applied. However, apoptosis of the drugged HepG2 cells appeared in later incubation phase with high dosage, and the apoptosis could be enhanced by supplemental dose of the intracellular polysaccharides. Nevertheless, the intracellular polysaccharides inhibited other human hepatocarcinoma cells such as BEL-7402 and Huh-7 but luckily stimulated human normal liver cell L02 only in a positive dose- and time-dependent manner; so did the sulfated extracellular polysaccharides when it inhibited HepG2 and L02 cells. However, the toxicity of sulfated extracellular polysaccharides to L02 cells can be eliminated by the intracellular polysaccharides.     

油菜田害虫调查及蚜虫防治药剂筛选

田间调查试验表明,合肥市油菜田害虫的发生种类有15种,主要害虫为蚜虫类、油菜潜叶蝇Phytomyza horticola、小菜蛾Plutella maculipennis和菜青虫Pieris rapae。其中,以蚜虫类危害最重,全生育期在冬、春季各有一个峰值。油菜苗期以萝卜蚜、小菜蛾危害较重,后期以桃蚜Myzus persicae、油菜潜叶蝇危害较重。蚜虫防治药剂筛选试验表明,70%吡虫啉水分散粒剂防治蚜虫药效高,持效期长,且对油菜安全,10%烯啶虫胺可溶液剂和25%吡蚜酮可湿性粉剂防效也较好,可作为70%吡虫啉水分散粒剂的轮换药剂。     

长江口棘头梅童鱼食物组成和摄食习性的季节变

棘头梅童鱼是长江口水域的常见小型经济鱼类,具有一定的捕捞价值.为了解长江口棘头梅童鱼食物组成和摄食习性的季节变化,本研究利用胃含物分析法对2013年11月—2014年8月在长江口水域捕获的棘头梅童鱼样本进行了分析.结果表明：长江口棘头梅童鱼全年摄食的饵料生物有8目30种,其中虾类(游泳亚目)是主要饵料生物,相对重要性指数百分比(IRI%)为38.5,优势指数(Ip)为79.1;其次为糠虾目和磷虾目.全年饵料生物中的主要优势种类为葛氏长臂虾、安氏白虾、脊尾白虾、长额刺糠虾、短额刺糠虾、光背节鞭水虱和中华哲水蚤等.饵料生物优势种存在季节差异,春季和夏季的主要优势种均为葛氏长臂虾、安氏白虾和短额刺糠虾,秋季主要优势种为长额刺糠虾、短额刺糠虾和脊尾白虾,冬季主要优势种为葛氏长臂虾、中华哲水蚤和中华假磷虾.长江口棘头梅童鱼全年空胃率为10.4%,冬季最高,秋季最低;全年平均胃饱满系数为0.6%,冬季最低,春季最高,摄食习性存在季节变化.     

不同盐胁迫对柳枝稷生物量、品质和光合生理的影响

为明确不同盐胁迫对柳枝稷生物量、品质及光合生理的影响,以无盐胁迫作为对照(CK),选取0.40%Na Cl、0.80%Na2SO4和0.80%Na HCO3进行了土柱试验。结果表明:(1)与CK相比,Na Cl、Na2SO4、Na HCO3胁迫下柳枝稷地上生物量、地下生物量、总生物量、籽粒产量及根冠比均显著降低(P0.05),总生物量分别降低49.39%、60.52%、76.45%,Na HCO3对柳枝稷的生长抑制作用最强,Na Cl最弱;(2)Na Cl胁迫下柳枝稷地上生物质灰分含量显著增高14.89%,Na2SO4胁迫下硫(S)含量显著增高262.32%,纤维素含量显著降低13.71%,Na HCO3胁迫下钾(K)含量显著增高54.95%,半纤维素含量显著增高10.87%,灰分和S含量的增高不利于柳枝稷地上生物质的燃烧利用,纤维素含量的降低和半纤维素含量的增高不利于其转化利用;(3)Na Cl、Na2SO4、Na HCO3胁迫下柳枝稷叶片净光合速率(Pn)分别显著降低21.89%、29.54%和24.59%,气孔限制因素可能是其光合作用受到抑制、生物量下降的关键因素。     

海滨滨麦叶片和根对不同厚度沙埋的生理响应差异分析

以烟台海岸抗风沙植物滨麦为研究材料,通过对不同厚度沙埋下其叶片和根部抗氧化酶活力(超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT))、丙二醛(MDA)含量和渗透调节物含量变化的分析,探讨了叶片和根部对沙埋生理响应的差异。试验按滨麦成株株高(约40 cm)对其进行了轻度沙埋(在株高1/4处)、中度沙埋(2/4处)和重度沙埋(3/4处)。在沙埋第6天,分别测定了不同厚度沙埋处理下,植株各段叶片和根抗氧化酶活力、MDA和渗透调节物含量。结果表明,轻度和中度沙埋均加速植株生长。与对照相比,经轻度、中度沙埋处理6 d,叶片平均MDA含量增加,在重度沙埋下降低。不同厚度沙埋6 d,叶片平均SOD活力和脯氨酸含量增加,而CAT活力、可溶性糖和可溶性蛋白质含量下降。但不同厚度沙埋均使沙上叶片MDA、脯氨酸、可溶性蛋白质含量和SOD和CAT活力增加,尤其是叶片顶部增加最为明显,使沙下叶片MDA、可溶性糖、可溶性蛋白质含量和CAT活力下降,导致同株沙上和沙下叶片MDA、脯氨酸、可溶性糖、可溶性蛋白质含量和SOD和CAT活力差异显著(P0.05)。与叶片相比,根中MDA、可溶性蛋白质含量和SOD和CAT活力较低,而POD活力和可溶性糖含量较高并与叶片差异显著(P0.05)。不同厚度沙埋6 d,滨麦根中MDA和可溶性蛋白质含量变化较小,可溶性糖含量和CAT、POD、SOD活力略有降低。研究表明,滨麦根和叶片对不同厚度沙埋的生理响应不同。沙埋直接作用于叶片并诱发叶内氧自由基积累,但叶片通过快速激活的抗氧化酶保护系统(CAT、SOD)维持氧自由基代谢平衡,以及渗透调节物(脯氨酸、可溶性糖)的积累维护细胞水分代谢平衡,并满足能量的需求和快速生长。但在不同厚度沙埋下,由于根系不受沙埋直接影响而生理变化较小,并且还维持较低的膜脂过氧化水平,这可能是根能维持正常的吸水输水功能并在沙埋处理过程中和沙埋后地上叶片快速生长摆脱沙埋的重要物质基础。     

菊黄东方鲀、暗纹东方鲀及其杂交F1代肌肉营养成分的比较分析

为了了解菊黄东方鲀(Takifugu flavidus)、暗纹东方鲀(T.obscurus)及其杂交F1代的肌肉营养特征,利用生物化学方法,从每类实验样本中取9尾对其肌肉中的粗蛋白、粗脂肪、水分、粗灰分和氨基酸成分进行了测定和分析。结果显示:(1)杂交F1代在生长方面具有明显的杂交优势,与亲本之间存在着显著差异(P0.05),杂交F1代的体重为其亲本的1.48~1.77倍;(2)杂交F1代肌肉水分含量与其母本含量相近,但粗脂肪含量均较亲本少(P0.05),粗蛋白含量则与亲本差异不显著(P0.05);(3)除色氨酸和胱氨酸外,16种氨基酸均在肌肉样本中被检测到,除甲硫氨酸外,其余15种氨基酸间含量均存在着显著性差异(P0.05)。菊黄东方鲀(♀)×暗纹东方鲀(♂)杂交F1代的总氨基酸含量最高,而暗纹东方鲀(♀)×菊黄东方鲀(♂)F1代总氨基酸含量则介于两亲本之间。对其必需氨基酸总量进行分析发现,菊黄东方鲀与其正反杂交F1代之间均存在显著性差异(P0.05),而暗纹东方鲀与其正反杂交F1代之间差异不显著(P0.05);(4)肌肉营养品质评价结果表明,菊黄东方鲀(♀)×暗纹东方鲀(♂)杂交F1代的鲜味氨基酸含量为26.68%,明显高于双亲样本(菊黄东方鲀22.28%、暗纹东方鲀25.20%),而暗纹东方鲀(♀)×菊黄东方鲀(♂)F1代的鲜味氨基酸总量(23.30%)较其父本偏高,但低于其母本。研究结果表明,杂交东方鲀的肌肉营养综合了双亲的优良特性,特别是是菊黄东方鲀(♀)×暗纹东方鲀(♂)杂交F1代,拥有最高的鲜味氨基酸含量,具有推广价值。     

Time to wake up: No impact of COMT Val158Met gene variation on circadian preferences,arousal regulation and sleep

Dopamine has been implicated in the regulation of sleep–wake states and the circadian rhythm. However, there is no consensus on the impact of two established dopaminergic gene variants: the catechol-O-methyltransferase Val158Met (COMT Val158Met; rs4680) and the dopamine D4 receptor Exon III variable-number-of-tandem-repeat polymorphism (DRD4 VNTR). Pursuing a multi-method approach, we examined their potential effects on circadian preferences, arousal regulation and sleep. Subjects underwent a 7-day actigraphy assessment (SenseWear Pro3), a 20-minute resting EEG (analyzed using VIGALL 2.0) and a body mass index (BMI) assessment. Further, they completed the Morningness–Eveningness Questionnaire (MEQ), the Epworth Sleepiness Scale (ESS) and the Pittsburgh Sleep Quality Index (PSQI). The sample comprised 4625 subjects (19–82 years) genotyped for COMT Val158Met, and 689 elderly subjects (64–82 years) genotyped for DRD4 VNTR. The number of subjects varied across phenotypes. Power calculations revealed a minimum required phenotypic variance explained by genotype ranging between 0.5% and 1.5% for COMT Val158Met and between 3.3% and 6.0% for DRD4 VNTR. Analyses did not reveal significant genotype effects on MEQ, ESS, PSQI, BMI, actigraphy and EEG variables. Additionally, we found no compelling evidence in sex- and age-stratified subsamples. Few associations surpassed the threshold of nominal significance (p < .05), providing some indication for a link between DRD4 VNTR and daytime sleepiness. Taken together, in light of the statistical power obtained in the present study, our data particularly suggest no impact of the COMT Val158Met polymorphism on circadian preferences, arousal regulation and sleep. The suggestive link between DRD4 VNTR and daytime sleepiness, on the other hand, might be worth investigation in a sample enriched with younger adults.     

Ca2+ Signaling and Cell Death Induced by Protriptyline in HepG2 Human Hepatoma Cells

The effect of protriptyline on Ca²⁺ physiology in human hepatoma is unclear. This study explored the effect of protriptyline on [Ca²⁺]_i and cytotoxicity in HepG2 human hepatoma cells. Protriptyline (50–150 μM) evoked [Ca²⁺]_i rises. The Ca²⁺ entry was inhibited by removal of Ca²⁺. Protriptyline‐induced Ca²⁺ entry was confirmed by Mn²⁺‐induced quench of fura‐2 fluorescence. Except nifedipine, econazole, SKF96365, GF109203X, and phorbol 12‐myristate 13 acetate did not inhibit Ca²⁺ entry. Treatment with the endoplasmic reticulum Ca²⁺ pump inhibitor 2,5‐di‐tert‐butylhydroquinone (BHQ) inhibited 40% of protriptyline‐induced response. Treatment with protriptyline abolished BHQ‐induced response. Inhibition of phospholipase C (PLC) suppressed protriptyline‐evoked response by 70%. At 20–40 μM, protriptyline killed cells which was not reversed by the Ca²⁺ chelator 1,2‐bis(2‐aminophenoxy)ethane‐N,N,N′,N′‐tetraacetic acid‐acetoxymethyl ester (BAPTA/AM). Together, in HepG2 cells, protriptyline induced [Ca²⁺]_i rises that involved Ca²⁺ entry through nifedipine‐sensitive Ca²⁺ channels and PLC‐dependent Ca²⁺ release from endoplasmic reticulum. Protriptyline induced Ca²⁺‐independent cell death.