家蚕微粒子病母蛾序贯抽样检验方法研究

本文根据统计学理论,设计了一套家蚕微粒子病母蛾序贯抽样检验方案,用此方案进行的抽样检验,不仅保证了将不合格蚕种判为合格蚕种的概率控制在1.5％以内,而且大大减少了将合格蚕种判为不合格蚕种的概率．并且镜检蛾数一般随总体病蛾率ρ而变化。当ρ较低或较高（相对于0.5％）时,需检验的数量就较少.本方案的实际检验工作量并未比现行检验方案有明显增加.     

高效液相色谱检测聚乙二醇化天花粉蛋白方法比较

目的：建立聚乙二醇化天花粉蛋白纯度的检测方法。方法：使用分子筛色谱法和反相高效液相色谱法测定了聚乙二醇天花粉蛋白。结果：聚乙二醇天花粉蛋白纯度分别为100％和98．14％。结论：高效液相色谱法简单易行、准确、灵敏度高,适用于聚乙二醇化天花粉蛋白纯度检测。     

基于混合效应模型和分位数回归的长白落叶松枝下高模型构建

枝下高是反映树冠特征的重要指标,准确预测枝下高对森林的经营管理和提高林分生产具有重要意义。本研究采用非线性回归构建枝下高广义基础模型,再进一步扩展到混合效应模型和分位数回归模型,通过“留一法”检验对模型的预测能力进行评价和比较。此外,使用4种抽样设计和不同抽样大小对枝下高模型进行校正,选择最佳的模型校正方案。结果表明：基于包含树高、胸径、林分每公顷断面积和优势木平均高的枝下高广义模型、扩展后的混合效应模型以及三分位数组合模型的预测精度均显著提高,混合效应模型略优于三分位数组合模型,最佳抽样校正方案为抽5株平均木。因此,推荐在实践应用中使用混合效应模型,抽5株样地平均木校正预测枝下高。     

m~*-v幂模型在检验昆虫种群空间分布中的应用

近年来,不少学者提出了各种检验昆虫种群空间分布的模型．本文提出一种新的模型：m~*-v幂模型──m~*＝avb,m~*与v呈幂关系．实例研究验证表明该模型具有适用范围更广和可行性大的优点．同时,根据m~*-v幂模型,推导出与其相配套的序贯抽样方程、最大抽样数量公式和最适抽样单元大小等公式．     

药物效用鉴定的计数二次抽样检验条件推断

本文给出了制定计数二次抽样检验方案,对未用的同类药品进行随机抽样,利用抽样结果,对剩下的N-n个样品的性能进行推断,得到判断药品是否失效的有效方法及相应的结论。     

四种主要大豆食叶害虫种群空间分布型及其应用研究

通过分层随机与连片调查获得114个样本．利用微机分别对豆天蛾卵和豆天蛾．银纹夜蛾．棉铃虫．豆灰蝶及混合种群的幼虫．进行了4种频次分布型检验和6项聚集度指标的测定．结果表明．上述害虫在豆田内均属零集分布．中分析了聚集原因．提出了“Z”字型10样点,每样点以1／3m双行为单位的抽样方法．确立了在两种允许误差下的抽样数量．进行了序贯抽样分析。     

竹小蜂幼虫空间分布型及抽样技术研究

竹小蜂幼虫空间分布型属于负二项分布;聚集度各指标测定结果表明,幼虫是聚集分布;经测定M^*=6．2836 0．3028^ˉx,屑于聚集分布。并用刀切法和F偏离度检验法对M^*进行了估计和检验测定为聚集分布。中分析并计算了林间调查的理论抽样数和序贯抽样决策,为今后该虫林间调查与防治提供了科学的依据。     

沙棘木蠹蛾的幼虫空间分布与抽样技术

利用常用的聚集度指标 ,对沙棘木蠹蛾HolcocerushippophaecolusHua ,Chou ,FangetChen幼虫的分布进行了分析。结果显示 ,该幼虫的空间分布格局为聚集型分布。通过Iwao的m - x回归检验方程 ,确定了其最适抽样数和最佳样方大小。采用对角线、之字型、五点式、棋盘式和平行线等 5种抽样方法对沙棘木蠹蛾幼虫的抽样技术分别进行了研究 ,结果表明 :最佳的抽样方法为棋盘式。     

我国HIV传播耐药监测中一类重要问题的处理（英文）

对于HIV传播耐药,WHO给出了截断序贯抽样方法来判别耐药水平,要求的最大样本量是47.但是,中国当前HIV流行水平较低,最大样本量很多时候无法达到,导致不能给出耐药水平的准确判断.为了解决这个问题,本文借助贝叶斯估计,给出了一种方法加于改进.借助于该方法,最大样本147被减小到38.做为该方法的应用,对于WHO抽样方法无发判断的新疆和湖南地区,我们分别给出了低水平耐药和中水平讨药的判定.同时,对于WHO抽样方法能够判断的地区,我们的方法也给出了相同的结论.本文的方法可以应用到样本量不够充足的地区,是WHO抽样方法的改进.     

桑蚕种样本孵化率偏差控制方法研究

桑蚕种孵化率是蚕种质量检验的重要指标,控制样本间孵化率偏差是保证孵化率准确性的关键．通过理论分析,给出了样本孵化率偏差的控制方法．在置信水平1-α下,两样本孵化率最大随机偏差约为u_(α/2)(p(1-p)(1/n_1 1/n_2))．并通过实际调查,与理论方法进行了具体分析与比较．并为蚕业生产提供了一套控制孵化率偏差的可行性方案．     

A new method of pebrine inspection of silkworm egg using multiprimer PCR

Using a mixture of several PCR primers, we evaluated whether multiprimer PCR is practically useful for the early and simultaneous detection of several kinds of microsporidia that cause silkworm pebrine. When genomic DNA extracted from silkworm eggs infected with Nosema bombycis was used as the DNA template, the specific DNA sequences were amplified by multiprimer PCR. In addition, similar results were obtained even when genomic DNA extracted from silkworms infected with N. bombycis was used as the DNA template. These findings suggest that multiprimer PCR using several primers designed for this study is practically useful for pebrine inspection of silkworm eggs.     

基于EB1基因的环介导等温扩增（LAMP）法检测感染家蚕微粒子病的蚕卵

【目的】家蚕 Bombyx mori 微粒子病一直影响着蚕种业的健康发展,快速而准确地检测出寄生于蚕卵中的家蚕微孢子虫 Nosema bombycis 对于有效控制家蚕微粒子病危害意义重大。【方法】环介导等温扩增（loop-mediated isothermal amplification, LAMP）法是一种快速、灵敏、特异的DNA体外恒温扩增方法,本文基于LAMP检测法的原理,依据家蚕微孢子虫孢子繁殖复制相关的 EB1 基因（GenBank登录号：KF421134.1）设计LAMP引物,对LAMP反应体系的最佳反应温度、内外引物浓度比、检测反应的特异性、灵敏度等进行研究,建立了一种检测蚕卵感染家蚕微粒子病的LAMP检测方法。【结果】结果表明,基于EB1 基因建立的LAMP检测方法在63℃恒温下在1.5 h内就可完成对样品的有效检测,本LAMP法对家蚕微孢子虫孢子DNA的检测灵敏度为5.0×10^-3 ng/μL,对EB1-pMD^TM19-T质粒标准品的检测灵敏度为1.0×10² copies/μL,同时对人工感染家蚕微粒子病单个母蛾产下蚕卵的1/8卵圈量或1粒蚕卵均能检出阳性结果。上述结果都分别应用凝胶电泳法、恒温荧光检测仪以及SYBR GreenⅠ显色肉眼观察法得到同步判定。【结论】本研究建立的基于EB1基因LAMP法可用于蚕卵微粒子病的检测,LAMP法为蚕种质量检验及成品卵微粒子病现场检疫提供了新技术。     

Genetic characterization of Iranian native Bombyx mori strains using amplified fragment length polymorphism markers

Genetic relationships within and among seven Iranian native silkworm strains was determined by DNA fingerprinting by using amplified fragment length polymorphism (AFLP) markers. In total, 189 informative AFLP markers were generated and analyzed. Estimates of Nei's gene diversity for all loci in individual strains showed a higher degree of genetic similarity within each studied strain. The highest and the least degrees of gene diversity were related to Khorasan Pink (h = 0.1804) and Baghdadi (h = 0.1412) strains, respectively. The unweighted pair-group method with arithmetic average dendrogram revealed seven strains of silkworm, Bombyx mori (L.), resolving into two major clusters. The highest degree of genetic similarity was related to Baghdadi and Harati White, and the least degree was related to Guilan Orange and Harati Yellow. The genetic similarity estimated within and among silkworms could be explained by the pedigrees, historical and geographical distribution of the strains, effective population size, inbreeding rate, selection intensity, and gene flow. This study revealed that the variability of DNA fingerprints within and among silkworm strains could provide an essential basis for breeders in planning crossbreeding strategies to produce potentially hetrotic hybrids in addition to contributing in conservation programs.     

Stimulation of replicative DNA synthesis by eukaryotic proteins bound to single-stranded DNA]

The paper deals with the effect of the single-strand (ss) DNA-binding proteins (SSB-proteins) from the Ehrlich ascites tumor (EAT) cells and from the eggs of silkworm, as well as the mouse serum blood proteins, having preferential affinity to ss DNA, on the DNA replicative synthesis in the EAT cells permeable for the macromolecules, and, for the silkworm proteins and on the DNA replicative synthesis in the nuclei from the eggs of silkworm proteins and on the DNA replicative synthesis in the nuclei from the eggs of silkworm permeable for macromolecules. SSB-proteins of EAT to considerable extent stimulated the DNA synthesis. At the same time, the other proteins (from the silkworm and from the serum) activated the DNA synthesis in the permeable cells to the less extent. It was found that SSB-proteins from the silkworm had a 1.5-13 fold stimulating effect on the DNA replicative synthesis in the homologous system (in the permeable nuclei). If the permeability for the macromolecules of the cells and nuclei treatment with Triton X-100 may be different, it is supposed that the activation of the DNA synthesis by the exogenous proteins depends on the homologous system of the DNA replicative complex. It is possible that the effect of the serum proteins on the DNA synthesis is connected with the masking of the ss regions of DNA which inhibited DNA-polymerase alpha. Perhaps the mechanisms of the activation of the DNA replicative synthesis by the proteins in vitro with the purified DNA polymerase alpha and in vivo are of different nature and are conditioned by homology of the deoxyribonucleoproteins.     

Kinetic effect of silkworm hemolymph on the delayed host cell death in an insect cell-baculovirus system

The kinetic effect of silkworm hemolymph on host cell viability during a baculovirus-induced insect cell death process was investigated. Host cell viability after viral infection is important for replication of the baculovirus DNA containing a recombinant gene and expression of the cloned gene. The baculovirus-induced insect cell death process can be divided into a delay phase and a first-order death phase, which are characterized by a delay time (t(d)) and a specific death rate (k(d)), respectively. For 0-10% silkworm hemolymph in the media, higher concentrations resulted in longer delay times and lower specific death rates. By adding 10% silkworm hemolymph, the delay time increased from 72 to 164 h, and the specific death rate was reduced from 13.8 x 10(-)(3) to 6.0 x 10(-)(3) h(-)(1). In addition, host cell viability correlated with DNA fragmentation, which is the biochemical hallmark of apoptosis. This indicates that the silkworm hemolymph inhibits the baculovirus-induced insect cell apoptosis. However, the silkworm hemolymph did not affect the number of hypothetical targets, which represents host cell susceptibility to the baculovirus. The concentration of fetal bovine serum (FBS) in the medium did not affect the delay time, while lower concentrations of silkworm hemolymph resulted in shorter delay times. This means that the substance which increases the longevity of the host cell is not in the FBS but in the silkworm hemolymph.     

超声造影和超声弹性成像在甲状腺良恶性结节鉴别诊断中的临床价值

目的:探讨超声造影与超声弹性成像鉴别诊断甲状腺良恶性结节的临床价值。方法:回顾性分析2011年1月-2013年6月我院经病理证实的128例甲状腺占位性病变患者(160个结节)的超声影像学资料,其中恶性结节68个,良性占位92个,评估实时超声造影与超声弹性成像诊断甲状腺良性与恶性结节的敏感性、特异性、准确率、阳性预测值及阴性预测值。结果:甲状腺良性结节超声造影检查以快进慢出、高增强为主;恶性结节以慢进快出、低增强为主。超声造影诊断甲状腺良、恶性结节的灵敏度、特异度与阳性预测值、阴性预测值及其诊断符合率分别为91.18%、92.39%、91.18%、93.41%、91.88%;超声弹性成像分别为89.71%、90.22%、87.14%、92.22%、90.00%,联合检查分别为94.12%、95.65%、94.12%、95.65%、95.00%,均高于常规超声的57.35%、72.83%、60.94%、69.79%、65.63%,比较差异有统计学意义(P0.05);联合检查灵敏度、符合率明显高于超声造影与超声弹性成像单一检查,比较差异有统计学意义(P0.05)。结论:超声造影与超声弹性成像在鉴别诊断甲状腺良恶性结节中均具有较高的应用价值,两种方法联合检查灵敏度及准确性更高。     

Silkworm hemolymph inhibits baculovirus-induced insect cell apoptosis

The effect of silkworm hemolymph on baculovirus-induced insect cell apoptosis was investigated. The addition of silkworm hemolymph into the culture medium either before or during the baculovirus infection increased the host cell longevity; however, its addition after the infection was less effective. This can be explained by the higher transfer rate of silkworm hemolymph which is caused by endocytosis during the virus internalization step. The delayed cell death due to silkworm hemolymph was not caused by an inhibition of the virus attachment and internalization steps. The apoptosis was analyzed using DNA fragmentation and TUNEL assays, and the resulting data confirm that silkworm hemolymph inhibits baculovirus-induced insect cell apoptosis.     

Introduction of foreign genes into silkworm eggs by electroporation and its application in transgenic vector test

Up to now, the most practical method for introducingforeign gene into insect eggs is the microinjection as thatwas utilized in Drosophila melanogaster [1–3]. The dis-covery of transposable elements including: P, Minos,Hermes, mariner, hobo, piggyBac, etc…     

基于RAPD分析的中国野桑蚕和家蚕遗传多样性和系统发育关系研究

对具代表性的中国11个地区的野桑蚕Bombyx mandarina进行了随机引物扩增多态性DNA(RAPD)分析,结果表明：不同地区的野桑蚕的遗传距离较大,最大为0.465(安康-镇江),最小的也有0.209(武汉-合肥);同一地区不同个体野桑蚕的遗传距离也较大,最大为0.318,最小的为0.144。它们均远大于家蚕B.mori品种内个体间的遗传距离(最大为0.068、最小为0.015),甚至超过了家蚕品种间的遗传距离(最大为0.258、最小为0.197)。这表明中国野桑蚕是一个十分混杂的、遗传多样性非常丰富的群体。另外,在大多数情况下野桑蚕的遗传距离表现出与空间距离正相关。遗传距离和系统发育分析结果显示陕西一带的野桑蚕成分复杂,有重要的演化意义。     

Microsatellite markers application on domesticated silkworm and wild silkworm

Abstract Twenty-seven sets of simple sequence repeat (SSR) primers were developed through hybridizing of (CA)n, (CT)n and (GT)n and sequencing the positive clones in libraries constructed by using p50 silkworm strain. Of those primer pairs, 26 sets of SSR primers amplified well in two regional wild silkworm strains. Ten domesticated silkworm strains and two regional wild silkworm strains were used for comparing the polymorphisms and for constructing a phylogenetic tree employing the UPGAM method. The result showed that the genetic distances within Japanese strains are closer than those of Chinese strains. And this result also implies that Japanese strains diverged from domesticated silkworm later than Chinese strains. According to the clustering result, the domesticated silkworm is firstly clustered in one class, but could be classified into two groups. Within a strain, the individual polymorphism of wild silkworm was significantly higher in abundance than those of domesticated silkworm. The S SR primers of domesticated silkworm could be used in genetic studies for wild silkworm.