摘要：
Objective: To investigate the reproductive toxicity and cytotoxicity of T-2 toxin, which is a mycotoxin, and to explore its potential apoptotic induction mechanism. Methods: Ovarian granulosa cells of rats were treated with T-2 toxin (1-100 nM) for 24 h. The cytotoxicity was assessed with MTT bioassay; apoptotic cells were identified microscopically by chromatin condensation and nuclear fragmentation with Hoechst 33258; mitochondrial membrane potential with hodamine 123 and reactive reactive oxygen species (ROS) with 2',7'-dichlorofluoresceinacetate (DCFH-DA) was analyzed by fluorometry; p53 and other apoptosis-related proteins such as Bax, Bcl-2, caspase-3, caspase-9 were determined by Western blot analysis, and related mRNA expressions were determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The caspase activity was measured by cleavage of the caspase substrate (Ac-DEVD-pNA for caspase-3. Ac-LEHD- pNA for caspase-9). Results: T-2 toxin inhibited the growth of granulosa cells in a concentration-dependent way. The result of Hoechst 33258 staining indicated that T-2 toxin induces granulosa cells apoptosis based on the typical apoptotic morphological changes. Subsequently, we found that T-2 toxin treatment induced ROS accumulation in granulosa cells, resulting in reduction of mitochondrial transmembrane potential. The induction of cell apoptosis was caused by the upregulation of p53, Bax, Bcl-2, Bax/Bcl-2 ration, and the activation of the caspases pathways. T-2 toxin-induced apoptotic granulosa cells significantly decreased through the use of antioxidant Trolox. Conclusion: These data suggest a possible underlying molecular mechanism for T-2 toxin that induces the apoptosis signaling pathway in rat granulosa cells by regulation of ROS-mediated mitochondrial pathway. (C) 2011 Elsevier Ireland Ltd. All rights reserved.
作者机构：
[Song, Hui-Qun; Huang, Si-Yang; Lin, Rui-Qing; Liu, Guo-Hua; Zhu, Xing-Quan; Tan, Hong-Wei] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, State Key Lab Vet Etiol Biol, Lanzhou, Gansu, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.;[Dong, Xia; Tan, Hong-Wei] Yunnan Agr Univ, Eastern Bee Res Inst, Kunming, Yunnan Prov, Peoples R China.;[Yuan, Zi-Guo; Lin, Rui-Qing] S China Agr Univ, Coll Vet Med, Guangzhou, Guangdong, Peoples R China.;[Zhao, Guang-Hui] NW A&F Univ, Coll Vet Med, Yangling, Shaanxi Prov, Peoples R China.
通讯机构：
[Tan, Hong-Wei] C;Chinese Acad Agr Sci, Lanzhou Vet Res Inst, State Key Lab Vet Etiol Biol, Lanzhou, Gansu, Peoples R China.
关键词：
Transfer RNA;Genomics;Invertebrate genomics;Hymenoptera;Honey bees;Ribosomal RNA;Mitochondria;Mitochondrial DNA
摘要：
In the present study, we determined the complete mitochondrial DNA (mtDNA) sequence of Apis cerana, the Asiatic cavity-nesting honeybee. We present here an analysis of features of its gene content and genome organization in comparison with Apis mellifera to assess the variation within the genus Apis and among main groups of Hymenoptera. The size of the entire mt genome of A. cerana is 15,895 bp, containing 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA (tRNA) genes and one control region. These genes are transcribed from both strands and have a nucleotide composition high in A and T. The contents of A+T of the complete genomes are 83.96% for A. cerana. The AT bias had a significant effect on both the codon usage pattern and amino acid composition of proteins. There are a total of 3672 codons in all 13 protein-coding genes, excluding termination codons. The most frequently used amino acid is Leu (15.52%), followed by Ile (12.85%), Phe (10.10%), Ser (9.15%) and Met (8.96%). Intergenic regions in the mt genome of A. cerana are 705 bp in total. The order and orientation of the gene arrangement pattern is identical to that of A. mellifera, except for the position of the tRNA-Ser(AGN) gene. Phylogenetic analyses using concatenated amino acid sequences of 13 protein-coding genes, with three different computational algorithms (NJ, MP and ML), all revealed two distinct groups with high statistical support, indicating that A. cerana and A. mellifera are two separate species, consistent with results of previous morphological and molecular studies. The complete mtDNA sequence of A. cerana provides additional genetic markers for studying population genetics, systematics and phylogeographics of honeybees.
摘要：
The prevalence of Enterococcus faecalis was investigated in diseased swine in Hunan Province, China between February 2007 and February 2011. A total of 385 diseased swine dissected in prosectorium from 10 representative administrative regions in Hunan Province were examined for the presence of E. faecalis. Forty-two out of 385 (10.91%) diseased swine were found to be infected with E. faecalis by bacteria isolations. E. faecalis was mainly recovered from the lung and joint cavity. Identification of the organisms was based on biochemical assays and sequencing of the partial 16S rRNA. The results of the present investigation have implications for the ongoing control of E. faecalis infections in swine and humans in Hunan Province, China.
通讯机构：
[Yuan, Hui] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
关键词：
DNA damage;Gossypol;Oxidase;Sertoli cells
摘要：
The study was designated to explore the toxic effects of gossypol on piglet sertoli cells. Sertoli cellswere isolated from piglet testes using a two-step enzyme digestion and followed by differential plating.Piglet sertoli cells were cultured and classified into five groups, that is, group A, the control withoutgossypol, group B with 2.5 μg/ml gossypol, group C with 5 μg/ml gossypol, group D with 10 μg/mlgossypol and group E with 20 μg/ml gossypol. We found that sertoli cells’ growth was inhibited bygossypol at dose 2.5 μg/ml when compared with the control group. The oxidase activity of sertoli cellalso decreased at 2.5 μg/m gossypol. Moreover, DNA damage of sertoli cells was observed at 5 μg/mlgossypol. Putting this into consideration, our study suggests that exposure of gossypol to sertoli cellsleads to an inhibition of sertoli cell growth and oxidase activity of sertoli cells at a low concentration,whereas gossypol results in DNA damage of sertoli cells at a higher concentration.Keywords: Gossypol, sertoli cells, oxidase, DNA damage
摘要：
The study was designed to explore the toxic effects of arsanilic acid on piglet Sertoli cells. Sertoli cells were isolated from piglet testes using a two-step enzyme digestion followed by differential plating. Piglet Sertoli cells were cultured and classified into the following five groups: group A, the control without arsanilic acid treatment; group B, cultured with 5 mu M arsanilic acid; group C, cultured with 50 mu M arsanilic acid; group D, cultured with 0.5 mM arsanilic acid; and group E, cultured with 5 mM arsanilic acid. We found that Sertoli cell growth was inhibited by arsanilic acid at 0.5 mM compared with the control, group A. The oxidase activity of Sertoli cells was decreased by arsanilic acid at 0.5 mM as evidenced by the observations that arsanilic acid increased MDA content but decreased the SOD and GSH-Px activities of Sertoli cells. Moreover, 50 mu M of arsanilic acid was observed to cause DNA damage in Sertoli cells. The results of our study suggest that exposure of Sertoli cells to arsanilic acid leads to induction of oxidative stress and inhibition of cell growth at a high concentration, while arsanilic acid causes DNA damage in Sertoli cells at a low concentration.
期刊：
Rapid Communications in Mass Spectrometry,2011年25(2):341-348 ISSN：0951-4198
通讯作者：
Yuan, Zong-Hui
作者机构：
[Chen, Dong-Mei; Wang, Xu; Liu, Zhao-Ying; Yuan, Zong-Hui; Tao, Yan-Fei] Huazhong Agr Univ, Coll Vet Med, HZAU MAO Key Lab Food Safety Evaluat, Natl Reference Lab Vet Drug Residues, Wuhan 430070, Hubei, Peoples R China.;[Liu, Zhao-Ying] Hunan Agr Univ, Fac Vet, Changsha 410128, Hunan, Peoples R China.;[Yuan, Zong-Hui] Huazhong Agr Univ, Coll Vet Med, HZAU MAO Key Lab Food Safety Evaluat, Natl Reference Lab Vet Drug Residues, Shizishan St, Wuhan 430070, Hubei, Peoples R China.
通讯机构：
[Yuan, Zong-Hui] H;Huazhong Agr Univ, Coll Vet Med, HZAU MAO Key Lab Food Safety Evaluat, Natl Reference Lab Vet Drug Residues, Shizishan St, Wuhan 430070, Hubei, Peoples R China.
摘要：
Carbadox (methyl-3-(2-quinoxalinylmethylene)-carbazate-N-1, N-4-dioxide) is a chemotherapeutic growth promoter added to feed for starter pigs. In this work, the metabolism of carbadox in rat, pig and chicken liver microsomes has been studied firstly. The incubation mixtures were then processed and analyzed for metabolites with a sensitive and reliable method based on high-performance liquid chromatography combined with hybrid ion trap/time-of-flight mass spectrometry (LC/MS-IT-TOF). With the help of chromatographic behavior and accurate mass measurements, it is possible to rapidly and reliably characterize the metabolites of carbadox. The structural elucidations of these metabolites were performed by comparing the changes in the accurate molecular masses and fragment ions generated from precursor ions with those of parent drug. The present results showed that the metabolism of carbadox in liver microsomes had qualitative species-difference. A total of seven metabolites were identified in rat liver microsomes. Five metabolites (Cb1-Cb3, Cb5, Cb7) were observed in pig and chicken liver microsomes. In addition, metabolite Cb6 was also detected in chicken liver microsomes. The peak areas of the metabolites in the three species are different. For the formations of Cb1, Cb2, Cb5 and Cb6, the rank order was rat>chicken>pig; Cb3; pig similar to chicken>rat. Cb1, Cb2 and Cb3 have been previously reported, whereas the other four metabolites were novel. The N -> O group reduction and hydroxylation followed by N -> O group reduction were the main metabolic pathways for carbadox in the three species. Copyright (C) 2010 John Wiley & Sons, Ltd.
