作者机构：
[Xu, Min-Jun; Liu, Guo-Hua; Zhu, Xing-Quan; Zhou, Dong-Hui] Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou, Gansu, Peoples R China.;[Liu, Guo-Hua; Zhu, Xing-Quan] Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.;[Gasser, Robin B.; Mohandas, Namitha] Univ Melbourne, Fac Vet Sci, Parkville, Vic 3052, Australia.;[Otranto, Domenico] Univ Bari, Dipartimento Sanita Pubbl & Zootecnia, Bari, Italy.;[Shen, Ji-Long] Anhui Med Univ, Dept Pathogen Biol, Hefei, Anhui, Peoples R China.
通讯机构：
[Liu, Guo-Hua] C;Chinese Acad Agr Sci, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou Vet Res Inst, Lanzhou, Gansu, Peoples R China.
摘要：
Human thelaziosis is an underestimated parasitic disease caused by Thelazia species (Spirurida: Thelaziidae). The oriental eyeworm, Thelazia callipaeda, infects a range of mammalian definitive hosts, including canids, felids and humans. Although this zoonotic parasite is of socio-economic significance in Asian countries, its genetics, epidemiology and biology are poorly understood. Mitochondrial (mt) DNA is known to provide useful genetic markers to underpin fundamental investigations, but no mt genome had been characterized for any members of the family Thelaziidae. In the present study, we sequenced and characterized the mt genome of T. callipaeda. This AT-rich (74.6%) mt genome (13,668 bp) is circular and contains 12 protein-coding genes, 22 transfer RNA genes and two ribosomal RNA genes, but lacks an atp8 gene. All protein-coding genes are transcribed in the same direction; the gene order is the same as those of Dirofilaria immitis and Setaria digitata (Onchocercidae), but distinct from Dracunculus medinensis (Dracunculidae) and Heliconema longissimum (Physalopteridae). Phylogenetic analyses of the concatenated amino acid sequence data for all 12 protein-coding genes by Bayesian inference (BI) showed that T. callipaeda (Thelaziidae) is related to the family Onchocercidae. This is the first mt genome of any member of the family Thelaziidae and should represent a new source of genetic markers for studying the epidemiology, ecology, population genetics and systematics of this parasite of humans and other mammals.
作者：
Wei, P.;Sun, E. C.;Liu, N. H.;Yang, T.;Xu, Q. Y.;...
期刊：
Archives of Virology,2013年158(5):1099-1104 ISSN：0304-8608
通讯作者：
Wu, D. L.
作者机构：
[Xu, Q. Y.; Sun, E. C.; Yang, T.; Wang, W. S.; Li, J. P.; Wu, D. L.; Qin, Y. L.; Feng, Y. F.; Liu, N. H.; Zhao, J.; Wei, P.; Zhang, C. Y.] Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Vet Biotechnol, Key Lab Vet Publ Hlth,Minist Agr, Harbin 150001, Peoples R China.;[Sun, E. C.; Wang, W. S.; Li, J. P.; Qin, Y. L.; Zhao, J.; Wei, P.] Chinese Acad Agr Sci, Grad Sch, Beijing 100081, Peoples R China.;[Yang, T.; Feng, Y. F.] Northeast Agr Univ, Coll Anim Med, Harbin 150030, Peoples R China.;[Zhang, C. Y.] HuNan Agr Univ, Coll Anim Med, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Wu, D. L.] C;Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Vet Biotechnol, Key Lab Vet Publ Hlth,Minist Agr, Harbin 150001, Peoples R China.
摘要：
Bluetongue virus (BTV) VP2 is an important antigenic protein that can be used for the differential diagnosis of different BTV serotypes. Here, we generated a serotype-specific monoclonal antibody (mab) against BTV1. A series of peptides synthesized based on the amino acid sequence of BTV1 VP2 were screened to define 115AQPLKVGL122 as the minimal linear peptide epitope recognized by mab 4B6. Using an immunofluorescence assay (IFA), we found that mab 4B6 reacted strongly with BTV1, but did not react with other BTV serotypes (BTV2-24). The 4B6 will serve as a novel reagent in the development of diagnostic tests for BTV1 infection.
摘要：
The barbel chub (Squaliobarbus curriculus), a kind of small commercial fish, is widespread in China. In this study, we sequenced the mitochondrial genome of the barbel chub from the Xiangjiang River. The total length of the mitochondrial genome is 16,619 bp, with the base composition of 31.19% A, 25.01% T, 27.68% C, and 16.12% G. The organization and arrangement of these genes are the same as that found in the teleosts, including 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes, and a major noncoding control region (D-loop region). Compared with the S. curriculus collected from Jiangsu province, there were 29 mutation sites in the mitogenome sequence of Xiangjiang S. curriculus. All the mutation sites were transitions and mainly occurred in protein-coding genes (21/29), two mutation sites occurred in transfer RNA, two occurred in ribosomal RNA, and four occurred in D-loop region. Among the 21 mutation sites in protein-coding genes, 6 mutation sites resulted in amino acid mutation in ND2, ATPase6, COX3, ND4, and Cytb genes, while the others were synonymous substitutions. These results indicated that there was genetic variation in different geographical populations of S. curriculus.
摘要：
Outbreaks of classical swine fever (CSF) have caused serious economic consequences in China. Phylogenetic analysis based on full-length E2 gene sequences showed that five classical swine fever virus (CSFV) isolates collected from Hunan province in 2011 and 2012, together with seven other isolates from neighboring provinces, Guangdong (5) and Guangxi (2), could be classified as a new subgenotype 2.1c, which may have been endemic in the south of China for at least fourteen years. Subgenotype 2.1c isolates share 90.2-94.9% and 89.9-93.8% nucleotide sequence similarity separately with those of subgenotype 2.1a and 2.1b in E2 gene, which are lower than the nucleotide identities between subgenotype 2.1a and 2.1b (91.1-95.7%). Further analysis based on a partial E2 gene sequence (216 nt) indicated that subgenotype 2.1c isolates are also circulating in Thailand. Alignment of E2 amino acid sequences showed that subgenotype 2.1c isolates exhibit a SPA -> TPV substitution at positions 777 and 779 compared with subgenotypes 2.1a and 2.1b. (C) 2013 Elsevier B.V. All rights reserved.
作者机构：
[Suo, Siqingaowa; Cao, Liyan; Ren, Yudong; Ding, Fan; Dou, Xiujing; Li, Guangxing; Zhu, Jiayi; Ren, Xiaofeng; Zhu, Weijuan] Northeast Agr Univ, Coll Vet Med, Harbin 150030, Peoples R China.;[Bu, Ri-e; Zhu, Jiayi; Ren, Xiaofeng] Inner Mongolia Univ Nationalities, Sch Life Sci, Tongliao, Inner Mongolia, Peoples R China.;[Yang, Qing] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Peoples R China.;[Zhao, Jianguo] Chinese Acad Sci, Inst Zool, State Key Lab Reprod Biol, Beijing 100101, Peoples R China.;[Ding, Fan] Chinese Armed Police Forest Force Command, Dept Hlth, Beijing 100097, Peoples R China.
通讯机构：
[Ren, Xiaofeng] N;Northeast Agr Univ, Coll Vet Med, 59 Mucai St, Harbin 150030, Peoples R China.
关键词：
PRV;VP6 gene;Cloning;Expression
摘要：
The context and purpose of the study included 1) bacterial expression of viral protein 6 (VP6) of porcine rotavirus (PRV) and generation of rabbit polyclonal antiserum to the VP6 protein; 3) establishment of a discrimination ELISA to distinguish PRV from a panel of other porcine viruses. The VP6 gene of PRV isolate DN30209 amplified by reverse transcription-PCR was 1356 bp containing a complete open reading frame (ORF) encoding 397 amino acids. Sequence comparison and phylogenetic analysis indicated that PRV DN30209 may belong to group A of rotavirus. Bacterially expressed VP6 was expressed in E.coli and anti-VP6 antibody was capable of distinguishing PRV from Porcine transmissible gastroenteritis virus, Porcine epidemic diarrhea virus, Porcine circovirus type II, Porcine reproductive and respiratory syndrome virus, Porcine pseudorabies virus and Porcine parvovirus. PRV VP6 expressed in E. coli can be used to generate antibodies in rabbit; anti-VP6 serum antibody can be used as good diagnostic reagents for detection of PRV.
摘要：
High-titer serologically detected male (SDM) antibody fragments are essential for specific binding to the SDM antigen and promoting its application. The A8 clone previously obtained from an original phage antibody library was further affinity-matured by light- and high-chain shuffling respectively, to generate the end product B9 clone. The binding capacity of B9 phage Fabs to male splenocytes doubled the value of its parental A8 clone (determined using ELISA). Based on immunofluorescent staining, B9-Fabs mainly bound to the surface antigen of male splenocytes and recognized testicular cells. The resulting B9-Fabs detected a single protein (approximately 40 kDa determined using Western blot analysis of male splenocytes and testis); its high SDM antigen binding ability might have been because of mutation sites and varied lengths of the amino acid sequences in the complementarity determining regions-3 of the kappa and Fd chains. The new recombinant clones of Fab that were phage-enhanced using chain shuffling were candidate molecules for investigating molecular mechanisms of SDM antigens specific binding and applications. (C) 2013 Elsevier Inc. All rights reserved.
摘要：
Objective: To investigate the effect of gossypol acetic acid (GA) on proliferation and apoptosis of macrophage cell line RAW264.7 and further understand the possible underlying mechanism responsible for GA-induced cell apoptosis. Methods: RAW264.7 cells were treated with GA (25-35μmol/L) for 24h. Concomitantly, the cytotoxicity was determined by using MTT assay, and apoptotic cells were identified by TUNEL assay, AO/EB staining and flow cytometry. Meanwhile, mitochondrial membrane potential (ΔΨm) with Rhodamine 123 and reactive oxygen species (ROS) with DCFH-DA were analyzed by fluorescence spectrofluorimeter, respectively. In addition, the expressions of caspase-3 and caspase-9 were assessed by Western Blot assay. Finally, the GA-induced cell apoptosis was evaluated by flow cytometry in the present of caspase inhibitors Z-VAD-FMK and Ac-LEHD-FMK , respectively. Results: GA significantly inhibited the proliferation of RAW264.7 cells in a dose-dependent manner, caused an obvious cell apoptosis and a loss of ΔΨm in RAW264.7 cells. Moreover, the ROS production in cells was elevated, and the levels of activated caspase-3 and caspase-9 were up-regulated in a dose-dependent manner. Notably, GA-induced cell apoptosis can be markedly inhibited by caspase inhibitors, respectively.Conclusion: These results suggest that GA-induced RAW264.7 cell apoptosis may be mediated via a caspase-dependent mitochondrial signaling pathway.
摘要：
This study investigated the inhibitory effects of sanguinarine (SA) on PKC-CPI-17 pathway in rat intestinal smooth muscle cells (ISMC). Previous studies indicate that the inhibitory effects of SA on ISMC contraction are possibly mediated by the Ca2+ influx. ISMC was treated with 1 mu M SA for 24 h remarkably inhibited the mRNA expression of m(2) and m(3) receptors. ISMC treated with 1 or 3 mu M SA for 30 min significantly decreased the mRNA expression of PKC-delta, PKC-epsilon, PKC-eta, and CPI-17. 1 mu M SA could markedly inhibit carbachol (CCh)-mediated increase PKC-delta, PKC-eta, and CPI-17 mRNA but had no effect in PKC-epsilon.Treatment of ISMC with SA (1 mu M, 30 min) caused a decrease in protein expression of PKC-delta. However, the expression of CPI-17 was significantly inhibited in a time-dependent manner. These results demonstrate that the inhibitory effect of SA is coupled with alteration of PKC-mediated signal transduction and intracellular Ca2+ concentration. (C) 2013 Elsevier Ltd. All rights reserved.
关键词：
Hybrid of Squaliobarbus curriculus (female) x Ctenopharyngodon idella (male);mitochondrial;genome
摘要：
In this work, we reported the complete mitochondrial DNA sequence of the hybrid of Squaliobarbus curriculus (female symbol) x Ctenopharyngodon idella (male symbol), which was obtained by artificial hybridization. The total length of the mitochondrial genome is 16,616 bp, with the base composition of 31.15% A, 25.02% T, 27.66% C, and 16.17% G. It contains 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes, and a major non-coding control region (D-loop region). The arrangement of these genes is the same as that found in the teleosts. All the protein initiation codons are ATG, except for COX1 that begins with GTG. The complete mitogenome of the hybrid of S. curriculus (female symbol) x C. idella (male symbol) provides an important data set for the study in genetic mechanism.
摘要：
Spirocerca lupi is a life-threating parasitic nematode of dogs that has a cosmopolitan distribution but is most prevalent in tropical and subtropical countries. Despite its veterinary importance in canids, the epidemiology, molecular ecology and population genetics of this parasite still remain unexplored. The complete mitochondrial (mt) genome of S. lupi was amplified in four overlapping long fragments using primers designed based on partial cox 1, rrn S, cox 2 and nad 2 sequences. Phylogenetic re-construction of 13 spirurid species (including S. lupi) was carried out using Bayesian inference (BI) based on concatenated amino acid sequence datasets. The complete mt genome sequence of S. lupi is 13,780 bp in length, including 12 protein-coding genes, 22 transfer RNA genes and two ribosomal RNA genes, but lacks the atp 8 gene. The gene arrangement is identical to that of Thelazia callipaeda (Thelaziidae) and Setaria digitata (Onchocercidae), but distinct from that of Dracunculus medinensis (Dracunculidae) and Heliconema longissimum (Physalopteridae). All genes are transcribed in the same direction and have a nucleotide composition high in A and T. The content of A + T is 73.73% for S. lupi, in accordance with mt genomes of other spirurid nematodes sequenced to date. Phylogenetic analyses using concatenated amino acid sequences of the 12 protein-coding genes by BI showed that the S. lupi (Thelaziidae) is closely related to the families Setariidae and Onchocercidae. The present study determined the complete mt genome sequence of S. lupi. These new mt genome dataset should provide novel mtDNA markers for studying the molecular epidemiology and population genetics of this parasite, and should have implications for the molecular diagnosis, prevention and control of spirocercosis in dogs and other canids.
作者机构：
[Gong, Qian Long; Xue, Li Qun; Wang, Nai Dong; Yu, Xing Long; Deng, Zhi Bang; Luo, Wei; Yuan, An Wen] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Xue, Li Qun] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.
关键词：
Porcine circovirus;PCV2b transmission;contact and vertical infection;Kunming mice
摘要：
To investigate porcine circovirus type 2b (PCV2b) transmission by contact and vertical infection in Kunming mice (an outbred mouse stock deriving from Swiss albino mice with a high ratio of gene heterozygosis), four mice in cage 6 were inoculated with PCV2b and 25 mice without any treatment were placed into cages 1 to 5 (five mice in each cage). Seven days after being infected, the PCV2-binoculated mice were co-mingled with non-inoculated mice from cages 1 to 5 successively at 7, 14, 21, 28 and 35 days post infection (dpi), respectively, for 3 days. In addition, eleven pregnant mice were injected with PCV2b. Samples were collected from non-inoculated mice and three newborn mice from each litter for PCV2b detection by polymerase chain reaction (PCR) and immunohistochemistry (IHC). The PCR results showed that PCV2b transmission rate among mice in cages 1, 2, 3, 4 and 5 was 0/5, 2/5, 5/5, 5/5 and 1/5, respectively. PCV2b antigen signals generally appeared in most organs of the non-inoculated mice in which viruses were detected by PCR. PCV2b DNA was also detected in newborn mice of PCV2b-infected litters, and viral antigen signals were observed in their organs as well. PCV2b was transmitted in Kunming mice by contact, and it also caused vertical infection through the placenta.
摘要：
Ticks are blood-sucking ectoparasites of great medical and veterinary significance that can transmit bacteria, protozoa, fungi and viruses, and cause a variety of human and animal diseases worldwide. In the present study, we sequenced the complete mitochondrial (mt) genome of Rhipicephalus sanguineus from China (RSC) and compared with that of R. sanguineus from USA (RSU). Nucleotide sequence difference in the full mt genome was 11.23% between RSC and RSU. For the 13 protein-coding genes, comparison revealed sequence divergences at both the nucleotide (9.34-15.65%) and amino acid (2.54-19.23%) levels between RSC and RSU. In addition, sequence comparison of the conserved mt cox1 and cytb genes among multiple individual R. sanguineus revealed substantial nucleotide differences between RSC and RSU but limited sequence variation within RSC. Phylogenetic analysis of ticks based on the amino acid sequence data of 13 protein-coding genes revealed that R. sanguineus from China and R. sanguineus from USA represent sister taxa (likely separate species). Taken together, the findings support the recently proposal that R. sanguineus tick may represents a species complex of at least two closely related species.
摘要：
Oesophagostomum spp., commonly known as 'nodule worms', is one of the important emerging nematode zoonoses worldwide. In the present study, we sequenced the complete mitochondrial (mt) genomes of two small ruminant nodule worms, Oesophagostomum asperum and Oesophagostomum columbianum, and compared them with that of pig nodule worms (Oesophagostomum dentatum and Oesophagostomum quadrispinulatum). The complete mt genomes of O. asperum and O. columbianum were 13,672 and 13,561 bp, respectively. Both mt genomes were circular, and consisted of 36 genes, including 12 genes coding for proteins, 2 genes for rRNA, and 22 genes for tRNA. The gene content and arrangement are identical to that of pig nodule worms. The availability of full mtDNA sequences of O. asperum and O. columbianum provide useful information for studying population genetics of Oesophagostomum spp., molecular epidemiology and control of O. asperum and O. columbianum infection in small ruminants. (C) 2013 Elsevier B. V. All rights reserved.
