酪蝇蛹对其成虫的引诱效果

室内采用Y型嗅觉仪测定酪蝇蛹、蛹壳及蛹的正己烷提取液对酪蝇成虫的引诱效果.结果表明:酪蝇的蛹、蛹的正己烷提取液对雌、雄成虫有明显的引诱,随着蛹量的增加其对雄虫的引诱效果增加,对雌虫的引诱效果下降;提取液浓度为5～12.5头当量/mL时,对雌、雄成虫有明显的引诱效果,但在浓度为2.5头当量/mL时对雄虫有明显的驱避效果,雌性蛹壳对两性成虫有明显的引诱效果,而雄性蛹壳只对雄虫有明显的引诱效果,对雌虫有明显的驱避效果.     

酶法制备海洋活性肽及其功能活性研究进展

海洋生物活性肽(Marine biological active peptide)是从海洋生物中提取的具有优化机体代谢环境、有益于机体健康的一类多肽。酶法制备海洋生物活性肽是目前最常用的制备方法,是通过适当的蛋白酶水解海洋生物蛋白来制备生物活性肽的一种方法。海洋生物活性肽在降血压、抗氧化、抗凝血及抗菌等方面效果显著,对治疗和预防疾病具有巨大潜力。介绍海洋生物活性肽在酶解制备及其生物学功能方面国内外研究进展,为进一步开展海洋活性多肽研究提供参考。     

异斑酷大蚕蛾幼虫、蛹和成虫的性别鉴定

本文描述了一种准确、快速鉴定异斑酷大蚕蛾末龄幼虫、蛹和成虫性别的方法。通过对蛹和成虫性别特征比较可知,雌虫生殖孔位于第8和第9腹节,且第8腹节不愈合,雄虫生殖孔仅位于第9腹节上,且生殖孔两侧有瘤状突起。雌性末龄幼虫头壳的蜕裂线附近浅色区域较多,雄性浅色区域较少,且雌性后唇基区域的颜色较雄性更浅。雌雄成虫差异较大,雄虫触角羽状,前翅一般无透明斑或透明斑很小;雌虫为近丝状的羽状触角,前翅有3个较大且连续排列的透明斑。经过对30对雌雄蛹的测量后,发现在蛹长、蛹宽与蛹重等体型数据上,雌雄蛹之间都存在显著性差异（P＜0.05）。     

云南保山地区两种胡蜂食用虫态营养成分分析及评价

为明确云南保山地区广泛分布和普遍食用的凹纹胡蜂（VespavelutinaaurariaSmith）和金环胡蜂（Vespa mandarinia Smith）的营养成分及差异,以野外采集的两种胡蜂为材料,采用常压直接干燥法、高温灼烧法、分光光度法等方法测定了凹纹胡蜂幼虫、预蛹和初蛹3个食用虫态及金环胡蜂初蛹虫态的水分、灰分、粗蛋白、氨基酸和总糖等营养成分含量,并对其进行分析和评价。结果表明,凹纹胡蜂幼虫、预蛹和初蛹3个虫态的水分含量分别为67.44%～75.71%、72.76%～73.28%、73.06%～74.20%;灰分含量分别为2.93%～3.77%、1.09%～2.38%、1.06%～2.57%;蛋白质含量分别为15.833%～17.326%、17.918%～18.471%、18.509%～19.529%;总糖含量分别为22.34%～25.44%、19.56%～21.19%、12.89%～19.43%;各食用虫态的营养成分含量与样品采样地和虫态关系密切,不同采样地的同一食用虫态或同一采样地的不同食用虫态的营养成分含量均具有显著差异。金环胡蜂初蛹的水分、灰分、蛋白质和总糖含量分别...     

抗肿瘤小分子多肽的研究进展

抗肿瘤小分子多肽具有分子量小、低毒性、高活性、易于穿透肿瘤细胞等特点,一些抗肿瘤小分子多肽已进入临床研究,成为肿瘤药物研发的新热点。本文从抗肿瘤小分子多肽的不同来源途径、结构改造及生物活性等方面,对其近年来的研究进展作简要概述。     

丝带凤蝶滞育与非滞育蛹及其成虫的形态学观察

丝带凤蝶Sericinus montelus是一种有开发价值的观赏昆虫,以蛹滞育越冬,成虫存在多型现象。本研究从体色、个体大小和蛹腹部刺突长度等方面比较了丝带凤蝶滞育与非滞育蛹及其成虫的形态差异。与非滞育蛹相比,滞育蛹体色较深,触角末端的淡黄色与体色差异明显,3日龄滞育蛹腹部第9节刺突长度是3日龄非滞育蛹的4倍左右,这些差异可以用于该虫蛹滞育早期判别。滞育蛹羽化成虫的翅展和尾突长度显著小于非滞育蛹羽化成虫,且腹部及翅面斑纹也存在明显的差异,这些差异与丝带凤蝶春型、夏型成虫的描述相一致,表明丝带凤蝶成虫季节多型是与滞育相关联的。     

沿海榆毡蚧的生物学特性及防治

园林重要害虫沿海榆毡蚧Eriococcus costatus(Danzig)在山西太谷1年发生1代,以2龄若虫及预蛹越冬。雌虫经卵、1龄若虫、2龄若虫、成虫完成生活史。雄虫则经卵、1龄若虫、2龄若虫、预蛹、冬前蛹(无翅型)、无翅雄成虫和部分若虫到冬后化蛹(有翅型)、有翅雄成虫,完成其生活史。影响该虫种群动态的主要生态因子包括温度、湿度、食料、天敌等,其中湿度在初孵若虫期是最主要的影响因子。4月上中旬为该虫的最佳防治时期,杀虫剂宜采用内吸性、触杀型的,且脂溶性效果较好。     

菜青虫不同虫态及虫龄的多酚氧化酶性质比较

初步比较了菜青虫Pieris rapae (L.)不同虫态及虫龄的多酚氧化酶(polyphenol oxidase)的性质。结果表明,不同虫态及虫龄的多酚氧化酶活力有很大的不同,其中以5龄幼虫酶活力最高,蛹酶活力最低。酶活力大小依次为： 5龄幼虫> 预蛹>4龄幼虫>3龄幼虫>蛹。以邻苯二酚为底物,研究了pH对多酚氧化酶活力的影响,结果表明酶催化底物氧化均有一个最适pH区域,不同虫态及虫龄的多酚氧化酶最适pH基本相同,其值为7.0±0.2 。不同虫态及虫龄的多酚氧化酶催化底物氧化的最适温度有很大的差异。3龄、4龄、5龄幼虫、预蛹和蛹的多酚氧化酶活力的最适温度分别为36.0±0.5℃、38.5±1.0℃、43.0±1.0℃、45.5±1.0℃和50.0±1.5℃,说明蛹期的多酚氧化酶活力的最适温度较高。进一步比较催化底物氧化反应的活化能,测得3龄、4龄、5龄幼虫、预蛹和蛹的多酚氧化酶活化能分别为：4 3.10±0.28、36.50±0.27、25.79±0.32、30.10±0.21和58.88±0.39 kJ/mol 。通过测定不同虫态及虫龄的多酚氧化酶催化邻苯二酚和L-多巴氧化反应的动力学特征参数,比较了不同虫期的酶对底物的亲和力。     

梨小食心虫蛹重对成虫繁殖力和寿命及下一代幼虫发育的影响

【目的】本文旨在明确营养状况不同造成的梨小食心虫Grapholitha molesta(Busck)雌、雄蛹重量差异对其羽化的成虫产卵量、产卵期、寿命及下一代(F1)幼虫发育的影响。【方法】室内条件下,通过不同的饲养方法,获得个体重量不同的梨小食心虫雌、雄蛹,待其羽化交配后,记录其产卵量、产卵时间和成虫寿命;卵孵化前后,分别测量卵和初孵幼虫大小,计算卵孵化率,统计幼虫发育历期。【结果】雌蛹重量对梨小食心虫的成虫产卵量影响显著,其重量与产卵量呈正相关(y=15.505x-59.292);同一条件下,雌蛹与雄蛹重量也呈正相关(y=0.823x-0.538)。同时,雌蛹重量对成虫产卵期影响也较大,蛹重大的个体羽化的雌虫比蛹重小的个体羽化的雌虫产卵高峰期提前1 d;较重、中等和较轻蛹羽化出的雌虫个体每天产卵量高于10粒/雌的时间分别为9~10,7和5~6 d;产卵量高于5粒/雌的时间分别为12~13,9和6~7 d。而雄蛹重量对产卵量、雄成虫寿命影响没有明显影响。较轻的蛹羽化的雌成虫寿命比较重蛹羽化的雌成虫短2~3 d;而雄蛹重量对其羽化的雄成虫寿命影响没有明显规律。雌、雄蛹重量对其羽化成虫的卵孵化率、卵和初孵幼虫的大小影响均不显著,对F_1幼虫发育历期影响也不显著。【结论】梨小食心虫雌蛹重对羽化成虫的产卵量和产卵期等影响显著,田间防治时应注意在不同条件下完成发育的个体,尤其是雌虫,由于营养差异引起的个体大小对随后种群增长的影响。     

第一代棉红铃虫幼虫化蛹习性初步观察

<正> 近几年,我们在开展棉红铃虫Pectinophoragossypiella(Saunders)综合防治研究的过程中,发现第一代红铃虫形成虫花后,除一部分幼虫留在虫花内化蛹外,相当数量的幼虫不知去向。查阅有关资料,多数记载第一代红铃虫幼虫老熟后一般留在虫花内化蛹;少数资料介绍,主要留在虫花内化蛹,少量也可入土化蛹。为此,我们从1981年开始,对第一代红铃虫幼虫化蛹习性作了初步观察。     

Baculoviral infection reduces the expression of four allergen proteins of silkworm pupa

Silkworm (Bombyx mori) larvae are widely used to express exogenous proteins. Moreover, some silkworm pupal proteins can be used as drug‐loading materials for selfexpressed oral tolerance drugs. However, several proteins expressed in silkworm pupae cause severe allergic reactions in humans and animals. Interestingly, some baculovirus vectors have been shown to alter the host gene and its expression in insect cells, but this has not been confirmed in silkworm. Here, we analyzed the effects of infection with an empty B. mori baculovirus (BmNPV) vector on silkworm pupal protein expression. Using a proteomics approach, the allergens thiol peroxiredoxin (Jafrac1), 27‐kDa glycoprotein (p27k), arginine kinase, and paramyosin as well as 32 additional differentially expressed proteins were identified. Downregulation of the messenger RNA expression of the four known allergens was observed after BmNPV infection; subsequent changes in protein expression were confirmed by the western blot analysis using polyclonal antibodies prepared with recombinant proteins of the four allergens. Collectively, these data indicate that the four known allergens of silkworm pupae can be reduced by infection ith an empty BmNPV vector to increase the safety of silkworm pupa‐based exogenous protein expression and drug delivery of oral pharmaceuticals. In addition, the four recombinant allergen proteins may contribute to the diagnosis of allergic diseases of silkworm pupa.     

培养基和栽培方式对蛹虫草子实体活性成分的影响

采用麦粒（W）、麦粒加蚕蛹粉（WW）、大米（R）、大米加蚕蛹粉（RW）的配料栽培方式,以及蚕蛹（CY）活体接种栽培方式培养蛹虫草子实体,比较蛹虫草子实体中多糖及核苷、游离糖醇和小分子糖类含量。结果表明：培养基和栽培方式影响蛹虫草多糖含量及其单糖组成和分子量分布,多糖含量最高的为蚕蛹活体接种培养的处理（CY）,多糖含量为6.19%,其他处理的多糖含量仅为CY的44.4%-62.8%;蚕蛹（CY）上培养的蛹虫草子实体中核苷类成分含量最高,在麦粒上培养获得的子实体中核苷含量显著高于在大米上的处理,并且麦粒添加蚕蛹粉后培养的子实体中尿苷、鸟苷、N⁶-(2-羟乙基)腺苷含量显著上升,大米添加蚕蛹粉后培养的子实体中尿苷、虫草素、N⁶-(2-羟乙基)腺苷含量显著上升;配料栽培的4个处理,其子实体中海藻糖含量为20.07%-23.40%,蚕蛹活体接种的处理（CY）海藻糖含量显著降低,为8.41%;添加蚕蛹粉后子实体中甘露醇含量显著降低。对各成分含量的数据进行归一化处理后进行蛹虫草品质的综合评价,在蚕蛹上培养的蛹虫草综合评分最高,麦粒为主要培养基培养的蛹虫草品质好于大米为主要培养基的,且添加蚕蛹粉的处理优于未添加的处理。     

Expression, purification and characterization of human GM-CSF using silkworm pupae (Bombyx mori) as a bioreactor

To date, many recombinant proteins have been expressed in Bombyx mori cells or silkworm larvae, apart from in pupae. Silkworm pupae may be more suitable for the expression of heterologous proteins as a bioreactor. If maintained at an appropriate temperature, silkworm pupae could be inoculated with recombinant baculovirus for the expression of a protein of interest. In this study, human granulocyte-macrophage colony-stimulating factor was successfully expressed in silkworm pupae using B. mori nucleopolyhedrovirus, purified and characterized with respect to its physico-chemical properties. The target protein expressed had an apparent molecular mass of 29 kDa and an isoelectric point of 5.1. The protein was purified using three chromatographic steps with a final recovery of 10.3%. Finally, approximately 3.5mg of the protein was obtained with a biological activity of up to 8.4 x 10(6) cfu mg(-1). The results of this study suggest that silkworm pupae represent a convenient and low-cost bioreactor for the expression of heterologous proteins.     

柞蚕核型多角体病毒基因表达载体系统的构建与基因表达

柞蚕是一种野外饲养的经济昆虫,主要分布在我国东北部山区。柞蚕以蛹滞育越冬,其蚕蛹个大、容易固定、保存时间长、无须饲养、容易运输等优点。利用柞蚕蛹作为生物反应器生产外来蛋白质,可以大规模机械化生产,减少操作上的烦琐和劳动力。本文利用柞蚕核型多角体病毒(AnpeNPV)作为基因表达载体,在柞蚕培养细胞(AnPe细胞)和柞蚕蛹中成功地表达了β-半乳糖苷酶基因（LacZ）,并利用AnPe细胞筛选、纯化获得了AnpeLacZ重组病毒。该重组病毒的β-半乳糖苷酶产量,在TC-100(含10%FBS)培养的AnPe细胞中最高酶活性为感染后第12天的40.9 units/ml,在SF-900Ⅱ培养的AnPe细胞中最高酶活性为感染后第18天的59.9 units/ml,后者表达量稍高,但时间滞后。AnpeLacZ在5℃保存了7个月的柞蚕蛹中,感染后第15天酶活性达到最高,雌蛹14.3 units/g,雄蛹11.7 units/g,雌蛹比雄蛹略高。结果显示,柞蚕核型多角体病毒和柞蚕蛹可以作为一个可以机械化大规模生产的新型杆状病毒基因表达载体系统开发和利用。     

Silkworm pupae powder ingestion increases fat metabolism in swim-trained rats

[Purpose]

Many researchers are trying to solve the metabolic syndrome by utilizing a variety of nutritional control and exercise. Of those, silkworm pupae peptides are known to inhibit the synthesis of fat. Therefore, we examine the effect of fat metabolism by supplying silkworm pupae (SP) for 5-week in swim-trained rats.

[Methods]

Animals were divided into four groups as a group (n = 32) fed a normal diet (CO) with exercise training (CE); a group fed a silkworm pupa diet (SPC) with an exercise training (SPE), respectively.

[Results]

Abdominal fat pads (abdominal and epididymal) weight were lowest in SPE. The serum triglyceride, total cholesterol concentrations were lower in the SP and the SPE. HDL-cholesterol, however, was not different between groups. Liver AMPK (AMP-activated protein kinase) was increased in the CE and the SPE. Liver PPAR-α (Peroxisome proliferator-activated receptor alpha) was increased in the SPC and SPE. L-FABP (liver fatty acids binding protein) was increased by SP ingestion. Liver CPT-1 (carnitine palmitoyltransferase-1) protein expression was increased by exercise training only.

[Conclusion]

In the present study showed that the silkworm pupae intake and/or swimming exercise training activates fat metabolism to reduce the concentration of serum lipids. Thus, the silkworm pupae intake leads to a reduction in fat storage, this is considered to be effective in the inhibition of the metabolic syndrome.     

Expression,purification and characterization of yeast protein disulfide isomerase produced by a recombinant baculovirus-mediated silkworm, <Emphasis Type="Italic">Bombyx mori</Emphasis>, pupae expression system

Protein disulfide isomerase (PDI) is a multifunctional polypeptide presents in the endoplasmic reticulum of the cell. Silkworm (Bombyx mori) pupae were used as hosts to produce recombinant PDI (rPDI). The concentration-dependent chaperone activity of rPDI was evidenced by the inhibition of the aggregation of rhodanese. Approximately 297 μg rPDI was purified from a single silkworm pupa. Results of rPDI treated with endoglycosidase H and N-glycanase, PNGase F, indicate that non-N-glycosylated rPDI (occupying 90%) and N-glycosylated rPDI are expressed in the silkworm expression system. The difference in glycosylation between silkworm pupae and yeast is discussed.     

Cuticle laccase of the silkworm,Bombyx mori: Purification,gene identification and presence of its inactive precursor in the cuticle.

Laccase is a multi-copper enzyme found in variety of organisms including plants, fungi and bacteria. In insects, laccase is thought to play an important role in cuticle sclerotization with its ability to catalyze the oxidation of phenolic compounds to their corresponding quinones. From the newly ecdysed pupae of the silkworm, Bombyx mori, we purified a dimer form of cuticular laccase with 70-kDa polypeptides. Mass spectrometric analysis of the tryptic fragments and cDNA sequence analysis revealed that the gene for the purified laccase (BmLaccase2) is an ortholog of laccase2, one of the multiple laccase genes found in insect genomes. BmLaccase2 is highly expressed in the epidermis prior to ecdysis, suggesting that the BmLaccase2 protein accumulates before ecdysis. However, the cuticle of newly ecdysed pupa does not have laccase activity, and the activity only becomes detectable several hours after ecdysis. These data suggest that cuticle laccase is synthesized as an inactive precursor, which is later activated after ecdysis. We also found that urea-solubilized cuticle protein extract contains an inactive form of laccase that can be activated by trypsin treatment.     

Purification and characterization of a novel immunomodulatory hexapeptide from alcalase hydrolysate of ultramicro-pretreated silkworm (Bombyx mori) pupa protein

Silkworm (Bombyx mori) pupa protein is one potential source of insect protein for use in food. Immunomodulatory peptides are specific protein fragments that can positively influence human health. Here, we purified a novel immunomodulatory hexapeptide from the alcalase hydrolysate of ultramicro-pretreated silkworm pupa proteinusing Sephadex gel filtration chromatography and reverse-phase high-performance liquid chromatography (RP-HPLC). The peptide sequence was determined by liquid chromatography–electrospray ionization– tandem mass spectrometry (LC-ESI-MS/MS). The results showed that the molecular mass of the purified peptide was 656.17 Da, and the amino acid sequence was Pro-Asn-Pro-Asn-Thr-Asn (PNPNTN). Splenocyte proliferation was 87.35% in the presence of 100 μg/ml of purified peptide. The splenocyte proliferation could be promoted upto 248.4% at 100 μg/ml of PNPNTN after induction by Concanavalin A (Con A). PNPNTN was stable in the presence of the gastrointestinal proteases pepsin and trypsin and at temperatures up to120°C. Taken together, these results show that this novel immunomodulatory hexapeptide from silkworm pupae has potential therapeutic value as an immunomodulatory component of functional food.     

Molecular characterization, tissue distribution, subcellular localization and actin-sequestering function of a thymosin protein from silkworm

We identified a novel gene encoding a Bombyx mori thymosin (BmTHY) protein from a cDNA library of silkworm pupae, which has an open reading frame (ORF) of 399 bp encoding 132 amino acids. It was found by bioinformatics that BmTHY gene consisted of three exons and two introns and BmTHY was highly homologous to thymosin betas (Tβ). BmTHY has a conserved motif LKHTET with only one amino acid difference from LKKTET, which is involved in Tβ binding to actin. A His-tagged BmTHY fusion protein (rBmTHY) with a molecular weight of approximately 18.4 kDa was expressed and purified to homogeneity. The purified fusion protein was used to produce anti-rBmTHY polyclonal antibodies in a New Zealand rabbit. Subcellular localization revealed that BmTHY can be found in both Bm5 cell (a silkworm ovary cell line) nucleus and cytoplasm but is primarily located in the nucleus. Western blotting and real-time RT-PCR showed that during silkworm developmental stages, BmTHY expression levels are highest in moth, followed by instar larvae, and are lowest in pupa and egg. BmTHY mRNA was universally distributed in most of fifth-instar larvae tissues (except testis). However, BmTHY was expressed in the head, ovary and epidermis during the larvae stage. BmTHY formed complexes with actin monomer, inhibited actin polymerization and cross-linked to actin. All the results indicated BmTHY might be an actin-sequestering protein and participate in silkworm development.     

Functional specificity of guanosine 3':5'-monophosphate-dependent and adenosine 3':5'-monophosphate-dependent protein kinases from silkworm.

Adenosine 3':5'-monophosphate-dependent protein kinase partially purified from silkworm pupae shows identical functional activities with those of mammalian protein kinases; the insect and mammalian kinases are completely exchangeable in the phosphorylation of muscle glycogen phosphorylase kinase and glycogen synthetase resulting in the activation and inactivation of the respective enzymes. In contrast, guanosine 3':5'-monophosphate-dependent protein kinase obtained from the same organism is totally inactive in this role and phosphorylates different, mainly seryl and some threonyl, residues of acceptor proteins. Substrates of the latter kinase intimately involved in the regulation of biological processes have remained unknown.