Determination of ethyl glucuronide in hair samples of Chinese people by protein precipitation (PPT) and large volume injection-gas chromatography-tandem mass spectrometry (LVI-GC/MS/MS)

Ethyl glucuronide (EtG) has been shown to be a suitable marker of excessive alcohol consumption. Determination of EtG in hair samples may help to differentiate social drinkers from alcoholics, and this testing can be widely used in forensic science, treatment programs, workplaces, military bases as well as driving ability test to provide legal proof of drinking. A method for determination of EtG in hair samples using large volume injection-gas chromatography-tandem mass spectrometry (LVI-GC/MS/MS) was developed and validated. Hair samples (in 1 mL deionized water) were ultrasonicated for 1h and incubated overnight; these samples were then deproteinated to remove impurities and derivatisated with 15 μL of pyridine and 30 μL of BSTFA. EtG was detected using GC/MS/MS in multiple-reaction monitoring mode. This method exhibited good linearity: y=0.0036 x+0.0437, R2=0.9993, the limit of detection and the limit of quantification were 5 pg/mg and 10 pg/mg, respectively. The extraction recoveries were more than 60%, and the inter-day and intra-day relative standard deviations (RSD) were less than 15%. This method has been applied to the analysis of EtG in hair samples from 21 Chinese subjects. The results for samples obtained from all of those who were teetotallers were negative, and the results for the other 15 samples ranged from 10 to 78 pg/mg, except for one negative sample. These data are the basis for interpretation of alcohol abuse.     

猴D型反转录病毒巢式PCR检测方法的建立

目的建立SRV-1巢式PCR检测方法并进行初步应用。方法针对SRV-1env基因的保守区序列,设计特异性引物,以感染SRV-1 Raji细胞提取出的含有前病毒DNA的基因组DNA为模板,进行巢式PCR反应。扩增产物测序后与GenBank报道的序列进行同源比对。将DNA样本进行10倍梯度稀释,以检测巢式PCR反应的灵敏度。使用该方法对正常Raji细胞以及感染SIV、STLV的外周血淋巴细胞DNA样本进行扩增,检测该方法的特异性。用建立的巢式PCR方法检测40份储存猴血标本。结果使用巢式PCR扩增出的特异片段经测序分析,结果证实与GenBank报道的序列一致。所建立的巢式PCR检测法检测限度可达1.5×10-3ng/μL,而且方法特异。用此方法检测40份猴血标本,未检测到阳性标本。结论初步建立SRV-1的巢式PCR检测方法,该方法灵敏、特异,为SRV-1的检测提供了一个快速、有效的手段。     

Changes in the biological activity of agropyrene on combined application with humus substances in the cultivation ofScenedesmus obliquus (TURP.) krüger

The work is concerned with the question of the effect of humic acids on the biological action of agropyrene, the effective substance from the rootstock of Agropyron repens (L.) P. BEAUV. The test organism was the alga Scenedesmus obliquus (TURP.) KRÜGER, cultivated in mineral solution. The basic biological test was the determination of the number and size of the cells of this alga. It was found that the way in which agropyrene acts on combined application with humus acids depends on which humus fraction is used. It can display a synergic action if humic acid is used, since this substance probably facilitates the penetration of agropyrene into the cells. On the other hand, fulvic acid counteracts the activity of agropyrene and this most probably by mutual binding and blocking of active groups and links.     

Calcium spikes in a leech nonspiking neuron

The NS neurons are nonspiking cells, present as pairs in each midbody ganglion of the leech nervous system, which display a very extensive arborization. They were shown to regulate the coactivation of motoneurons. Here we have investigated the electrophysiological properties of these neurons under the hypothesis that transmission along the extensive neurites requires the aid of voltage-dependent conductances. The results indicate that NS neurons respond to electrical stimulation with a spike-like event, which was not an all-or-none but rather a graded phenomenon that depended on the intensity and duration of the electrical stimulus. The spike-like response was activated at a membrane potential of approximately −50 mV; its amplitude was a logarithmic function of the extracellular Ca²⁺ concentration and was unaffected by a broad range of changes in the extracellular Na⁺ concentration; intracellular application of tetraethylammonium (TEA) caused a large increase in its amplitude and duration. These data indicate that NS neurons bear voltage-dependent low-threshold Ca²⁺ and TEA-sensitive K⁺ conductances that could contribute to shaping synaptic signals, or transmission along the extensive neuritic tree.     

Coexistence of diploid, triploid and tetraploid crucian carp (Carassius auratus) in natural waters

Background

Crucian carp (abbreviated CC) belongs to the genus of Carassius within the family of Cyprinidae. It has been one of the most important freshwater species for Chinese aquaculture and is especially abundant in the Dongting water system of Hunan province. CC used to be considered as all diploid forms. However, coexistence of diploid (abbreviated 2nCC), triploid (abbreviated 3nCC) and tetraploid crucian carp (abbreviated 4nCC) population of the Dongting water system was first found by our recently researches.

Results

We examined the ploidy level and compared biological characteristics in different ploidy CC. In reproductive mode, 2nCC was bisexual generative and 4nCC generated all-female offspring by gynogenesis. However, 3nCC generated progenies in two different ways. 3nCC produced bisexual triploid offspring fertilized with 3nCC spermatozoa, while it produced all-female triploid offspring by gynogenesis when its ova were activated by heterogenous spermatozoa. The complete mitochondrial DNA of three different ploidy fishes was sequenced and analyzed, suggesting no significant differences. Interestingly, microchromosomes were found only in 3nCC, which were concluded to be the result of hybridization. Allogenetic DNA fragments of Sox genes were obtained in 3nCC and 4nCC, which were absent in 2nCC. Phylogenetics analysis based on Sox4 gene indicated 3nCC and 4nCC formed a separate group from 2nCC.

Conclusions

In summary, this is the first report of the co-existence of three types of different ploidy crucian carps in natural waters in China. It was proved that the coexistence of different ploidy CC was reproductively maintained. We further hypothesized that 3nCC and 4nCC were allopolyploids that resulted from hybridization. The different ploidy CC population we obtained in this study possesses great significance for the study of polyploidization and the evolution of vertebrates.     

葡萄Alfin like转录因子家族的鉴定和表达分析

Alfin like (AL) 转录因子家族对高盐、低温、干旱等非生物胁迫反应具有重要的调控作用。该研究采用同源比对的方法检索鉴定葡萄AL转录因子家族基因、分析其生物信息学特性,并采用qRT PCR方法分析非生物胁迫下AL基因的表达特征,以探究葡萄AL基因在非生物逆境胁迫中的功能。 结果表明：(1)在葡萄中共鉴定出6个AL基因家族成员,分别命名为VvAL1~VvAL6,且6个成员分别分布在6条染色体上。(2)葡萄中AL转录因子具有高度保守的DUF3594结构域和PHD结构域,各家族成员均含有5个外显子和4个内含子;上游启动子区域分析发现大量植物激素与非生物胁迫响应相关的顺式作用元件。(3)基因芯片表达模式分析显示,盐、干旱、ABA胁迫以及低温(5 ℃)处理均显著影响葡萄AL家族基因(VvAL1~VvAL6)的表达。(4)qRT PCR检测显示,不同胁迫处理下AL基因在葡萄叶片中的表达水平不同;ABA处理下葡萄AL转录因子家族基因表达量较对照均显著下调,但在PEG处理下差异不显著;在盐胁迫处理下,VvAL2、VvAL4、VvAL5基因的表达量均显著上调,分别是对照的23倍、8.5倍和10.5倍,而VvAL1和VvAL6基因的表达量均显著下调,分别是对照的33倍和25倍。研究发现,葡萄AL转录因子家族与植物激素和非生物胁迫密切相关,尤其是该家族基因强烈响应高盐胁迫。     

"观察SO2对植物的影响"实验的改进

介绍了对人民教育出版社的高中生物学必修教材第2册103页,实验12“观察二氧化硫对植物的影响”的部分改进。利用洗净的金龙鱼牌5L塑料油瓶和洗净的空矿泉水瓶等废旧物资进行实验操作。材料易于寻找,成本低廉,培养了学生“变废为宝”的节约思想;用NaOH溶液吸收装置内的SO2,避免了环境污染,培养了学生的环保意识。     

The genome of the marine medaka Oryzias melastigma

Marine medaka (Oryzias melastigma) is considered to be a useful fish model for marine and estuarine ecotoxicology studies and has good potential for field‐based population genomics because of its geographical distribution in Asian estuarine and coastal areas. In this study, we present the first whole‐genome draft of O. melastigma. The genome assembly consists of 8,602 scaffolds (N50 = 23.737 Mb) and a total genome length of 779.4 Mb. A total of 23,528 genes were predicted, and 12,670 gene families shared with three teleost species (Japanese medaka, mangrove killifish and zebrafish) were identified. Genome analyses revealed that the O. melastigma genome is highly heterozygous and contains a large number of repeat sequences. This assembly represents a useful genomic resource for fish scientists.