亚致死浓度溴虫腈和毒死蜱对等钳蠊螨生长繁殖和解毒酶的影响

【目的】害虫综合治理中,化学防治对天敌资源也具有一定的杀伤力。为缓解这一矛盾,本研究旨在探究溴虫腈和毒死蜱亚致死浓度处理对等钳蠊螨Blattisocius dentriticus生长繁殖及解毒酶活性和基因表达量的影响。【方法】利用药膜法处理2-3日龄等钳蠊螨成螨24 h,测定溴虫腈和毒死蜱的亚致死剂量;测定和比较LC_(10)和LC_(30)剂量这两种药剂处理后F_0和F_1代的产卵量、产卵期、卵孵化率和雌成螨寿命等生物学特性的变化;通过酶活力分析和RT-PCR分别测定LC_(10), LC_(30)和LC_(50)剂量下这两种药剂处理后钳蠊螨成螨体内谷胱甘肽-S-转移酶(glutathione S-transferase, GST)、细胞色素P450(cytochrome P450, CYP450)、羧酸酯酶(carboxylesterase,CarE)酶活力及其基因表达量变化。【结果】溴虫腈和毒死蜱处理24 h对等钳蠊螨成螨的LC_(50)分别为42.56 mg/L和72.42 mg/L。溴虫腈和毒死蜱LC_(10)和LC_(30)剂量处理等钳蠊螨雌成螨后,与对照(清水处理)相比,仅F_0代雌成螨寿命和产卵期显著缩短(P0.05),而产卵量和卵孵化率无明变化。酶活力测定结果发现,GST, CYP450和CarE的活力在溴虫腈和毒死蜱LC_(10)和LC_(30)剂量处理后无明显变化,而LC_(50)剂量下,上述3种酶的活力均显著增加(P0.05)。基因表达结果表明,溴虫腈LC_(10), LC_(30)和LC_(50)剂量处理下2个GST基因(BdGST3和BdGST6)、3个CYP450基因(BdCYP2-4)和5个CarE基因(BdCarE1-5)表达均显著上调;在毒死蜱这3个剂量处理下3个GST基因(BdGST1,BdGST3和BdGST4)、3个CYP450基因(BdCYP2,BdCYP5和BdCYP6)和2个CarE基因(BdCarE1和BdCarE2)表达量均显著上调。【结论】结果表明,LC_(10)和LC_(30)剂量的溴虫腈和毒死蜱亚致死剂量会抑制F_0代雌成螨的生长繁殖;LC_(10), LC_(30)和LC_(50)剂量下这两种药剂可诱导等钳蠊螨GST, CarE和CYP450基因表达;LC_(50)剂量能明显诱导等钳蠊螨体内GST, CarE和CYP450活性上升。该研究为等钳蠊螨抗性品系的筛选及田间应用提供了理论依据。     

嘌呤拟似物对豚鼠胃窦环行肌自发性收缩及电活动的影响

为探讨非肾上腺素能非胆碱能神经递质对胃窦环行肌功能的调节作用,在离体胃平滑肌上观察了嘌呤拟似物对胃窦环行肌自发性收缩活动和电活动的影响。电活动用传统的细胞内微电极记录,并和收缩活动同步描记于多道生理记录仪。结果表明,嘌呤能P2Y受体激动剂,三磷酸腺苷(ATP)和2-methylthio ATP(2-MeSATP)均增强胃窦平滑肌的收缩活动,但不影响电活动,而且ATP和2-MeSATP对胃平滑肌收缩活动的增强作用可被嘌呤能P2Y受体阻断剂,reactive blue-2和苏拉明(suramin)所阻断。用100μmol／L α,β-MeATP引起的脱敏感使P2X受体被阻断,ATP增强胃窦平滑肌收缩活动的效应不受影响。嘌呤能P2X受体激动剂,α,β-MeATP明显抑制胃窦环行肌自发性收缩活动,同时使膜电位明显超极化。ATP对胃窦平滑肌的收缩作用不被L型钙通道阻断剂尼卡地平(nicardipine)阻断,但细胞外用无钙液灌流时这种效应则完全被阻断。用前列腺素合成抑制剂消炎痛预先处理20min后,ATP和2-MeSATP仍能增强胃窦平滑肌的自发性收缩活动。以上结果提示：(1)ATP和2-MeSATP通过嘌呤能P2Y受体增强胃窦平滑肌的自发性收缩活动,而α,β-MeATP或β,γ-MeATP通过嘌呤能P2X受体使膜电位超极化,引起胃窦平滑肌的舒张;(2)ATP和2-MeSATP增强胃窦平滑肌自发性收缩活动的效应依赖于细胞外钙,但钙离子进入细胞的途径并不是电压依赖性钙通道;(3)ATP和2-MeSATP增强胃窦平滑肌自发性收缩活动的效应不通过前列腺素介导。     

SAHA Inhibits Somatic Hyperalgesia Induced by Stress Combined with Orofacial Inflammation Through Targeting Different Spinal 5-HT Receptor Subtypes

Neurochemical Research - Epigenetic regulation of gene expression has been implicated in the development of chronic pain. However, little is known about whether this regulation is involved in the...     

Electrochemical aptasensor using the tripropylamine oxidation to probe intramolecular displacement between target and complementary nucleotide for protein array

Tripropylamine (TPA) has different oxidation efficiency at double stranded (ds)-and single stranded (ss)-DNA-modified electrodes. Using this property, a simple but sensitive biosensor using TPA oxidation to probe the intramolecular displacement was constructed with the analysis of lysozyme as model for the first time. After the complementary ss-DNA strand of anti-lysozyme aptamer was immobilized onto gold electrode via gold-thiol bond, the incubation with the aptamer resulted in the formation of ds-DNA. Lysozyme (in 10 μL sample) binding with aptamer displaced the complementary strand because of the high affinity of lysozyme and its aptamer, corresponding to the dissociation of the ds-DNA. The modified electrode was swept in 20mM TPA solution from 0.2 to 0.95 V. The difference in oxidation current was used to quantify the content of lysozyme with a linear range from 1.0 pM to 1.1 nM. That means 10 amol or 6.0 × 10(6) lysozyme molecules can be detected. Because the signal is produced from the preconcentrated TPA at the electrode surface, the high sensitivity is achieved over the single site labelling strategy. The proposed method is simple, stable, specific, and time-saving while the complicated sample pre-treatment and the labelling to the DNA strand are avoided. The biosensor was validated by the analysis of the diluted egg white sample directly. The recovery and reproducibility were 93.3-100% and 1.4-4.2%, respectively.     

Retracted: Long noncoding RNA MEG3 inhibits proliferation and migration but induces autophagy by regulation of Sirt7 and PI3K/AKT/mTOR pathway in glioma cells

Glioma is a common primary brain tumor with high mortality rate and poor prognosis. Long noncoding RNA maternally expressed gene 3 (MEG3) is a tumor suppressor in diverse cancer types. However, the role of MEG3 in glioma remains unclear. We aimed to explore the effects of MEG3 on U251 cells as well as the underlying mechanisms. U251 cells were stably transfected with different recombined plasmids to overexpress or silence MEG3. Effects of aberrantly expressed MEG3 on cell viability, migration, apoptosis, expressions of apoptosis-associated and autophagy-associated proteins, and phosphorylated levels of key kinases in the PI3K/AKT/mTOR pathway were all evaluated. Then, messenger RNA (mRNA) and protein expression of Sirt7 in cells abnormally expressing MEG3 were estimated. In addition, effects of abnormally expressed MEG3 and Sirt7 on U251 cells were determined to reveal the underlying mechanism of MEG3-associated modulation. Cell viability and migration were significantly reduced by MEG3 overexpression whereas cell apoptosis as well as Bax and cleaved caspase-3/-9 proteins were obviously induced. Beclin-1 and LC3-II/LC3-I were upregulated and p62 was downregulated in MEG3 overexpressed cells. In addition, the autophagy pharmacological inhibitor (3-methyladenine, 3-MA) affected the effect of MEG3 overexpression on cell proliferation. Furthermore, the phosphorylated levels of key kinases in the PI3K/AKT/mTOR pathway were all reduced by MEG3 overexpression. Sirt7 was positively regulated by MEG3 expression, and effects of MEG3 overexpression on U251 cells were ameliorated by Sirt7 silence. MEG3 suppressed cell proliferation and migration but promoted autophagy in U251 cells through positively regulating Sirt7, involving in the inhibition of the PI3K/AKT/mTOR pathway.     

高山植物黄管秦艽cDNA 文库构建与表达序列标签(EST) 分析

黄管秦艽( Gentiana officinalis) 是一种重要的藏药高山植物, 本研究构建了该物种开花期的cDNA 文库。经检测达到中等cDNA 文库水平, 文库滴度为1 . 2×107 pfu􊄯ml , 重组率95.9% , 插入片段平均长度大于500 bp。对343 个随机挑选的重组克隆进行部分测序, 获得的ESTs 经编辑后共有181 条有效序列。经生物信息学方法分析181 条表达序列标签(EST) 代表144 个单克隆序列, 其中55 个与已鉴定的基因同源, 35 个序列与未鉴定的EST 匹配, 54 个未找到同源序列; 后两者共有89 个EST 序列未发现功能相似的蛋白。对已鉴定的EST进行功能分析发现, 相关基因主要编码以下蛋白: 与蛋白表达相关的占35%; 光合作用相关的占22%; 新陈代谢相关的占18%; 抗性相关的占11%; 质膜运输和细胞分裂相关的分别占5% ; 染色体变化和细胞信号转导的分别占2%。根据有效EST 序列设计引物, 通过RT-PCR 进一验证了所得EST 的准确性。这些研究结果为将来研究黄管秦艽的功能基因以及该物种与相关物种的群体遗传学、进化生物学等方面提供了基础。     

Plane of vertebral movement eliciting muscle lengthening history in the low back influences the decrease in muscle spindle responsiveness of the cat

Proprioceptive feedback is thought to play a significant role in controlling both lumbopelvic and intervertebral orientations. In the lumbar spine, a vertebra's positional history along the dorsal-ventral axis has been shown to alter the position, movement, and velocity sensitivity of muscle spindles in the multifidus and longissimus muscles. These effects appear due to muscle history. Because spinal motion segments have up to 6 degrees of freedom for movement, we were interested in whether the axis along which the history is applied differentially affects paraspinal muscle spindles. We tested the null hypothesis that the loading axis, which creates a vertebra's positional history, has no effect on a lumbar muscle spindle's subsequent response to vertebral position or movement. Identical displacements were applied along three orthogonal axes directly at the L(6) spinous process using a feedback motor system under displacement control. Single-unit nerve activity was recorded from 60 muscle spindle afferents in teased filaments from L(6) dorsal rootlets innervating intact longissimus or multifidus muscles of deeply anesthetized cats. Muscle lengthening histories along the caudal-cranial and dorsal-ventral axis, compared with the left-right axis, produced significantly greater reductions in spindle responses to vertebral position and movement. The spinal anatomy suggested that the effect of a lengthening history is greatest when that history had occurred along an axis lying within the anatomical plane of the facet joint. Speculation is made that the interaction between normal spinal mechanics and the inherent thixotropic property of muscle spindles poses a challenge for feedback and feedforward motor control of the lumbar spine.     

Isolation and characterization of microsatellite DNA primers in Juniperus przewalskii Kom (Cupressaceae)

Juniperus przewalskii (Cupressaceae) is a dominant tree species endemic to the northeast Qinghai-Tibetan Plateau. This species plays an important role in maintaining the arid ecosystem in this region. However, natural distributions of this species have been declined. In order to develop effective conservation methods, it is important to know the distribution of the genetic diversity within and among populations. In this study, we developed nine new microsatellite loci for this species. We used the combining biotin capture method to enrich AG/CT/AC/GGT microsatellites. The polymorphisms of each locus were further assessed in 12 individuals from four geographically distant populations. The number of alleles per locus varied from three to six and expected heterozygosity ranged from 0.58 to 0.70. These loci together provide a useful tool to investigate the genetic diversity of this species. In addition, all markers have been crossly checked in the other four congeneric species.     

Development of 10 microsatellite loci for <Emphasis Type="Italic">Rheum tanguticum</Emphasis> (Polygonaceae)

Rhubarb is an important Traditional Chinese Medicine. However, the wild resource has been declining. In order to design appropriate conservation methods for the official species across their natural distributions, it is important to characterize their genetic diversity. Here, we describe the development of 10 new microsatellite loci for AC/TG/CCA in Rheum tanguticum. The microsatellites were enriched using the combined biotin capture method. The polymorphism of each locus was further assessed in 12 individuals from four geographically distinct populations of this species. The number of alleles ranged from three to seven and the expected heterozygosity ranged from 0.53 to 0.73. All markers have been checked in the other three species in the genus and two of them together comprise the official medicinal rhubarb resource with R. tanguticum. These microsatellite markers could provide a useful tool for genetic and conservation studies of the rhubarb species.