HCV CE2融合蛋白在家蚕培养细胞、蚕幼虫中的表达及其初步应用

目的:高效表达HCV CE2融合蛋白并进行初步应用. 方法:将2a型HCV全长CE2基因的cDNA重组于质粒pBacPAK8,构建重组转移载体pBacPAK-CE2,与经线性化修饰的家蚕核型多角体病毒(BmNPV)DNA共转染家蚕培养细胞株,构建重组病毒. 双酶切及PCR鉴定重组病毒基因组中目的片段的表达. 重组病毒感染BmN细胞株及五龄家蚕幼虫后,SDS-PAGE分析细胞培养上清、细胞抽提物及幼虫体液样品中,HCV CE2融合蛋白的特异性条带. 并用间接ELISA法初步检测表达产物的生物活性. 结果:双酶切及PCR鉴定重组病毒基因组含有约1.6 kb的目的片段,重组病毒感染后的BmN细胞培养上清、细胞抽提物及幼虫体液样品中,均可见一Mr约90×103的特异性条带;用间接ELISA检测证明表达产物具有较好的免疫原性. 结论:HCV CE2融合基因在家蚕培养细胞及蚕幼虫中获得了高效表达,并具有生物活性,为进一步进行疫苗的研究及临床诊断试剂的开发奠定基础.     

Cutaneous larva migrans with folliculitis: a new clinical presentation of this infestation

Two patients (a 35-year-old woman and a 33-year-old man, just returned from trips to Jamaica and Barbados, respectively) presented with cutaneous larva migrans with folliculitis, a rare and atypical clinical presentation of this infestation. Histopathological examination revealed a follicular and perifollicular infiltrate prevalently consisting of lymphocytes and neutrophils, with numerous eosinophils. Both patients were successfully treated with oral albendazole.     

Photochemical effect of silver nanoparticles on flesh fly larval biological system

With the progress of nanoscience and its applications, silver nanoparticles (AgNPs) have become one of the most interesting nanoparticles owing to their use in different fields. However, the excessive use of AgNPs and its products may cause toxicity in both the environment and in human health. The main goal of this research is to study the toxic and photochemical effects of AgNPs against Sarcophaga argyrostoma larvae through ultrastructure, morphological change, and DNA damage. Treating midgut epithelium with AgNPs led to many alterations in dark conditions, disintegrated epithelium, swollen cells, and shrunken nucleus. Organelles appeared in a loose manner and mitochondria were without cristae, endoplasmic reticulum had dark spots, and peritrophic membrane was loose in appearance. Fatty tissues were vacuolized and muscle fibers lacked normal striations and had many gaps and lysosomal bodies. In the light conditions, the epithelium appeared with detached cells and many vacuoles, organelles were ruptured with many gaps in between, and secretory vesicles were scattered. Peritrophic membrane disappeared. Muscles collapsed and vacuolized loosed fatty tissues were detected. On the other hand, control larvae epithelium appeared regularly distinct, with organelles intact and muscles had clear normal striations. Data showed that AgNPs caused ultrastructural and morphological changes of the external cuticle of the 4th instar larvae along with a significant effect on DNA damage that occurred after the larval treatment, reflecting the toxicity of AgNPs.     

外界条件对虫草菌有性型发生的影响

采用不同的温度,保湿和光照等处理,对自然界中已感染亚香棒虫草菌的湖南棒蝠蛾进行培养试验。结果表明,给予一定的外界条件,能促使虫草正常生长发育并诱发菌僵虫形成有性型。     

灭幼脲对淡色库蚊幼虫发育的影响

在淡色库蚊发育过程中,用半数致死浓度的灭幼脲处理其二令幼虫,让其发育到四令,再用双向电泳分析受处理的幼虫蛋白和正常二令、四令幼虫蛋白。经比较发现受处理的幼虫的蛋白组份明显少于正常;其多肽点分布与正常相似,但不同于二令幼虫。这提示灭幼脲能抑制幼虫蛋白合成及其生长,但似乎不能抑制其发育。     

吸水链霉菌应城变种种内融合子FR－008产生的新抗生素灭蚊幼的研究

本文报道吸水链霉菌应城变种种内融合子ＦＲ－００８产生的新抗生素（简称ＦＲ－００８抗生素）对３种蚊虫幼虫的灭蚊活性以及蚊龄、温度、紫外线对ＦＲ－００８抗生素灭蚊活性的影响。实验证明：ＦＲ－００８抗生素对３种蚊虫幼虫均有较强的致死作用。对３龄致倦库蚊、白纹伊蚊和中华按蚊幼虫的２４ｈＬＤ＿（５０）分别为１．３３０９μｇ／ｍｌ、２．０１１３μｇ／ｍｌ和２．８３１７μｇ／ｍｌ。不同虫龄对ＦＲ－００８抗生素的敏感性不同，１龄至２龄幼虫比４龄初幼虫敏感。ＦＲ－００８抗生素，在１９℃～３１℃之间，随温度升高灭蚊效果更好。紫外线照射可显著降低ＦＲ－００８抗生素的灭蚊活性。     

以旋毛虫成虫感染小鼠的研究

本文以旋毛虫成虫为感染期感染小鼠获得成功。用２００条旋毛虫成虫经口感染小鼠，在鼠的肠内发现成虫，肌肉内发现幼虫。实验结果显示，２日龄成虫感染的１０只鼠肠内幼虫数６～５７条，平均３０．８条，肌肉内幼虫数１７～７３条，平均３８．１条。７日龄成虫感染的１０只鼠肠内检获成虫数２～３４条，平均１８．４条，肌肉内幼虫数０～５５条，平均２５．４条。１５日龄成虫感染的１０只鼠肠内成虫数０～８条，平均２；２条２只鼠的肌肉内有幼虫，分别是２和３条。３０日龄成虫感染的小鼠肠内和肌肉内未见成虫和幼虫。因此，可以认为在旋毛虫生活史过程中，旋毛虫成虫亦能起感染期作用感染宿主。     

INAUGURAL ARTICLE by a Recently Elected Academy Member:Candidate ionotropic taste receptors in the Drosophila larva

We examine in Drosophila a group of ∼35 ionotropic receptors (IRs), the IR20a clade, about which remarkably little is known. Of 28 genes analyzed, GAL4 drivers representing 11 showed expression in the larva. Eight drivers labeled neurons of the pharynx, a taste organ, and three labeled neurons of the body wall that may be chemosensory. Expression was not observed in neurons of one taste organ, the terminal organ, although these neurons express many drivers of the Gr (Gustatory receptor) family. For most drivers of the IR20a clade, we observed expression in a single pair of cells in the animal, with limited coexpression, and only a fraction of pharyngeal neurons are labeled. The organization of IR20a clade expression thus appears different from the organization of the Gr family or the Odor receptor (Or) family in the larva. A remarkable feature of the larval pharynx is that some of its organs are incorporated into the adult pharynx, and several drivers of this clade are expressed in the pharynx of both larvae and adults. Different IR drivers show different developmental dynamics across the larval stages, either increasing or decreasing. Among neurons expressing drivers in the pharynx, two projection patterns can be distinguished in the CNS. Neurons exhibiting these two kinds of projection patterns may activate different circuits, possibly signaling the presence of cues with different valence. Taken together, the simplest interpretation of our results is that the IR20a clade encodes a class of larval taste receptors.Olfaction and taste are mediated by receptors of widely diverse families (1, 2). Studies of receptor expression have been critical to our understanding of chemosensory perception. Historically, the identification of several classes of receptors has been based largely on their expression patterns, with functional validation not becoming available until years later. Studies of receptor expression have informed our understanding of the principles of chemosensory coding. In some cases, analysis of receptor expression has suggested, and subsequently revealed, complex and elegant mechanisms of receptor gene regulation. Finally, in many cases, elucidation of receptor expression patterns has allowed chemosensory stimuli of particular ecological, evolutionary, or behavioral significance to be assigned to individual receptors.The Drosophila larva offers major advantages as an organism in which to study the molecular and cellular basis of taste. The larval taste system is relatively simple and can be investigated with incisive molecular and genetic approaches. Understanding the molecular and cellular mechanisms by which Drosophila larvae evaluate potential food sources may suggest means of manipulating the feeding of other insect larvae, some of which consume agricultural crops and collectively cause immense damage to the world’s agricultural output (3).The head of the Drosophila larva contains three external chemosensory organs (4) (Fig. 1). The dorsal organ (DO) is innervated by the dendrites of 21 olfactory neurons and nine gustatory neurons. The terminal organ (TO) and ventral organ contain the dendrites of ∼21 and approximately seven gustatory neurons, respectively.Open in a separate windowFig. 1.Chemosensory organs in the larval head and pharynx. VO, ventral organ. We have depicted the VPS as anterior to the DPS, but they are close and their apparent relative positions depend on the viewing angle. The DPO is more difficult to identify than the other organs, and its position relative to the DPS and PPS may depend on the larval stage; we have not depicted neural processes for it.There are also internal chemosensory organs lining the pharynx, each existing as a bilaterally symmetrical pair: the dorsal, ventral, and posterior pharyngeal sensilla (DPS, VPS, and PPS, respectively) (5–7) (Fig. 1). Each organ contains ∼17, 16, and 6 neurons, respectively, most of which are likely to be gustatory (5). Another organ, the dorsal pharyngeal organ (DPO), contains five neurons (3, 5, 8). A variety of other neurons in the body wall of the thorax and abdomen, and at the posterior tip of the larva, are also likely to be chemosensory (9–11).The Gustatory receptor (Gr) family comprises 60 genes (12, 13). Expression analysis of the Gr genes using the GAL4-UAS system has shown that 39 of the predicted proteins are likely to be expressed in the TO, DPS, VPS, or PPS of the larva (11, 14, 15). However, a receptor-to-neuron map of the TO neurons suggested that many TO neurons did not express any Gr genes, consistent with the notion that some larval taste neurons may express other kinds of taste receptors (15).The Ionotropic receptor (IR) family comprises 60 genes, of which members of one clade encode odor receptors (16). Another clade of 35 IR genes, called the IR20a clade, was recently shown to be expressed in gustatory neurons of Drosophila adults (17). Analysis of GAL4 drivers of 28 genes of the clade revealed expression of 16 drivers in adult taste neurons, collectively representing all taste organs of the fly. Virtually nothing is known of their expression in larvae.Here, we carry out a systematic expression analysis of the IR20a clade in the larval gustatory system. We find that 11 of the GAL4 drivers show expression in larval gustatory organs. Seven drivers are expressed in the DPS, with different drivers expressing in different DPS neurons, and one of these drivers is also expressed in the VPS. Another driver is expressed in the DPO; another is expressed in nonneuronal cells of the TO; and three are expressed in the body wall, where they are associated with sensory hairs, sensory cones, and trachea. The neurons that express the drivers show different projection patterns in the larval CNS. Some drivers show dynamic expression patterns over the course of development. The simplest interpretation of the results is that the IR20a clade encodes a class of larval taste receptors.     

松鼠单蚤亚洲亚种三龄幼虫形态描述

对松鼠单蚤亚洲亚种（Ｍｏｎｏｐｓｙｌｌｕｓ ｓｃｉｕｒｏｒｕｍ ａｓｉａｔｉｃｕｓ）三种幼虫的形态进行描述。大鄂８齿,前头刚毛６对,后头刚毛６对。自第一胸节至第９腹节主刚毛列毛数依次为８（２）,８（２）,８（２）,１０（２）,１０（２）,１０（２）,１０（２）,１０（２）,１０（２）,１０（２）和１２根。第１０腹节肛梳２例,前列８－１２根后列１４根。肛柱宽短,末端钝圆,基部有８或９根支柱毛。与同