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| 1 | Acrylamide exposure impairs blood-cerebrospinal fluid barrier function显示文摘Previous studies show that chronic acrylamide exposure leads to central and peripheral neuropathy. However, the underlying mechanisms remained unclear. In this study, we examined the permeability of the blood-cerebrospinal fluid barrier, and its ability to secrete transthyretin and transport leptin of rats exposed to acrylamide for 7, 14, 21 or 28 days. Transthyretin levels in cerebrospinal fluid began to decline on day 7 after acrylamide exposure. The sodium fluorescein level in cerebrospinal fluid was increased on day 14 after exposure. Evans blue concentration in cerebrospinal fluid was increased and the cerebrospinal fluid/serum leptin ratio was decreased on days 21 and 28 after exposure. In comparison, the cerebrospinal fluid/serum albumin ratio was increased on day 28 after exposure. Our findings show that acrylamide exposure damages the blood-cerebrospinal fluid barrier and impairs secretory and transport functions. These changes may underlie acrylamide-induced neurotoxicity. | Xue Yao Licheng Yan Lin Yao Weijun Guan Fanxu Zeng Fuyuan Cao Yanshu Zhang | 2014 | Neural Regeneration Research2014，9，5： | 4 |
| 2 | Establishment of cell lines with porcine spermatogonial stem cell properties显示文摘Background:Spermatogonial stem cells(SSCs)are capable of both self-renewal and differentiation to mature functional spermatozoa,being the only adult stem cells in the males that can transmit genetic information to the next generation.Porcine SSCs hold great value in transgenic pig production and in establishment of porcine models for regenerative medicine.However,studies and applications of porcine SSCs have been greatly hampered by the low number of SSCs in the testis as well as the lack of an ideal stable long-term culture system to propagate porcine SSCs perpetually.Results:In the present study,by lentiviral transduction of plasmids expressing the simian virus 40(SV40)large T antigen into porcine primary SSCs,we developed two immortalized cell lines with porcine SSC attributes.The established cell lines,with the expression of porcine SSC and germ cell markers UCHL1,PLZF,THY1,VASA and DAZL,could respond to retinoic acid(RA),and could colonize the recipient mouse testis without tumor formation after transplantation.The cell lines displayed infinite proliferation potential,and have now been cultured for more than 7 months and passaged for over 35 times without morphological abnormalities.Conclusions:We have for the first time established porcine SSC lines that could provide abundant cell sources for mechanistic studies on porcine SSC self-renewal and differentiation,thereby facilitating development of an optimal long-term culture system for porcine primary SSCs and their application to animal husbandry and medicine. | Yi Zheng Tongying Feng Pengfei Zhang Peipei Lei Fuyuan Li Wenxian Zeng | 2020 | Journal of Animal Science and Biotechnology2020，11，3： | 2 |
| 3 | Single-cell RNA-sequencing reveals the dynamic process and novel markers in porcine spermatogenesis显示文摘Background:Spermatogenesis is the process by which male gametes are formed from spermatogonial stem cells and it is essential for the reliable transmission of genetic information between generations.To date,the dynamic transcriptional changes of defined populations of male germ cells in pigs have not been reported.Results:To characterize the atlas of porcine spermatogenesis,we profiled the transcriptomes of~16,966 testicular cells from a 150-day-old pig testis through single-cell RNA-sequencing(scRNA-seq).The scRNA-seq analysis identified spermatogonia,spermatocytes,spermatids and three somatic cell types in porcine testes.The functional enrichment analysis demonstrated that these cell types played diverse roles in porcine spermatogenesis.The accuracy of the defined porcine germ cell types was further validated by comparing the data from scRNA-seq with those from bulk RNA-seq.Since we delineated four distinct spermatogonial subsets,we further identified CD99 and PODXL2 as novel cell surface markers for undifferentiated and differentiating spermatogonia,respectively.Conclusions:The present study has for the first time analyzed the transcriptome of male germ cells and somatic cells in porcine testes through scRNA-seq.Four subsets of spermatogonia were identified and two novel cell surface markers were discovered,which would be helpful for studies on spermatogonial differentiation in pigs.The datasets offer valuable information on porcine spermatogenesis,and pave the way for identification of key molecular markers involved in development of male germ cells. | Lingkai Zhang Fuyuan Li Peipei Lei Ming Guo Ruifang Liu Ling Wang Taiyong Yu Yinghua Lv Tao Zhang Wenxian Zeng Hongzhao Lu Yi Zheng | 2022 | Journal of Animal Science and Biotechnology2022，13，2： | 2 |
| 4 | Transcriptome-wide Dynamics of m^(6)A mRNA Methylation During Porcine Spermatogenesis显示文摘Spermatogenesis is a continual process that occurs in the testes,in which diploid spermatogonial stem cells(SSCs)differentiate and generate haploid spermatozoa.This highly efficient and intricate process is orchestrated at multiple levels.N^(6)-methyladenosine(m^(6)A),an epigenetic modification prevalent in mRNAs,is implicated in the transcriptional regulation during spermatogenesis.However,the dynamics of m^(6)A modification in non-rodent mammalian species remains unclear.Here,we systematically investigated the profile and role of m^(6)A during spermatogenesis in pigs.By analyzing the transcriptomic distribution of m^(6)A in spermatogonia,spermatocytes,and round spermatids,we identified a globally conserved m^(6)A pattern between porcine and murine genes with spermatogenic function.We found that m^(6)A was enriched in a group of genes that specifically encode the metabolic enzymes and regulators.In addition,transcriptomes in porcine male germ cells could be subjected to the m^(6)A modification.Our data show that m^(6)A plays the regulatory roles during spermatogenesis in pigs,which is similar to that in mice.Illustrations of this point are three genes(SETDB1,FOXO1,and FOXO3)that are crucial to the determination of the fate of SSCs.To the best of our knowledge,this study for the first time uncovers the expression profile and role of m^(6)A during spermatogenesis in large animals and provides insights into the intricate transcriptional regulation underlying the lifelong male fertility in non-rodent mammalian species. | Zidong Liu Xiaoxu Chen Pengfei Zhang Fuyuan Li Lingkai Zhang Xueliang Li Tao Huang Yi Zheng Taiyong Yu Tao Zhang Wenxian Zeng Hongzhao Lu Yinghua Lv | 2023 | Genomics, Proteomics & Bioinformatics2023，21，4： | 1 |
| 5 | Molecular Cloning and Bioinformatics Analysis of crp Gene in Vibrio alginolyticus显示文摘[Objectives]The crp gene in Vibrio alginolyticus was studied.[Methods]The crp gene of V.alginolyticus HY9901 was cloned and analyzed by bioinformatics.[Results]The ORF of the crp gene is 633 bp long and the predicted amino acid sequence encompasses 210 amino acid residues.The physicochemical property analysis indicated that the chemical formula of CRP is C_(1051)H_(1704)N_(290)O_(308)S_(10) with a molecular weight of 23.6514 kDa,and its theoretical pI is 7.74.Besides,the protein is stable and hydrophilic.The protein had three protein kinase C phosphorylation site,three casein kinase II phosphorylation site,one N-terminal myristoylation site.The BLAST analysis on the sequence revealed high homology with CRPs in other Vibrio species,and particularly the sequence shares about a homology of 99.52%with the CRP in V.parahaemolyticus.The SWISS-MODEL software simulated the subunit tertiary structural model of the CRP,and the similarity with template 3hif.1.A was 95.71%.[Conclusions]This study provides a reference for the search for efficient protective antigens against vibrosis. | Zhiqing WEI Weijie ZHANG Linlin YIN Haiyun FENG Junlin WANG Fuyuan ZENG Xing XIAO Huanying PANG | 2022 | Agricultural Biotechnology2022，11，2： | 0 |
| 6 | Bioinformatics Analysis of DNA-binding Response Regulator PhoP in Vibrio alginolyticus显示文摘[Objectives]To amplify the DNA-binding response regulator PhoP in Vibrio alginolyticus and analyze its sequence characteristics and subunit structure.[Methods]According to the sequence of the DNA-binding response regulator PhoP in V.alginolyticus,a pair of specific primers was designed for PCR amplification,and the bioinformatics of the sequence amplified was analyzed.Using MEGA 5.0 software,the phoP phylogenetic tree was constructed by the neighbor-joining method.Using SWISS-MODEL software,the three-dimensional structural model of the PhoP subunit was simulated.[Results]The full-length phoP gene was 732 bp,encoding a total of 243 amino acids.The predicted theoretical molecular weight of the protein is about 27.67 kD,and the isoelectric point is 5.09.The prediction results of protein subcellular localization,SignalP 4.0,TMHMM Server 2.0 and SoftBerry-Psite show that PhoP is located in the cytoplasm,and is stable and hydrophobic;there is a signal peptide cleavage site between amino acids 29 and 30,and there is no transmembrane region.The amino acid sequence contains one Asn-glycosylation site,one protein kinase C phosphorylation site,seven casein kinase II phosphorylation sites,one tyrosine kinase phosphorylation site,three myristoylation sites,and seven C-terminal microbody targeting signal sites.The PhoP of V.alginolyticus has high homology with that of Vibrio campbellii.The PhoP subunit of V.alginolyticus has similar configuration to the single-subunit RegX3 protein of Mycobacterium tuberculosis.[Conclusions]This study has a positive effect on the prevention and control of vibriosis and the improvement of the current aquatic economic animal breeding environment. | Yifan WU Junlin WANG Fuyuan ZENG Weijie ZHANG Xing XIAO Huanying PANG | 2022 | Asian Agricultural Research2022，14，3： | 0 |
| 7 | Cloning and Bioinformatics Analysis of TpiA Gene of Vibrio alginolyticus HY9901显示文摘[Objectives]To clone and analyze the TpiA gene of Vibrio alginolyticus HY9901.[Methods]According to the TpiA gene sequence of V.alginolyticus,a pair of specific primers was designed,and its full length was amplified by PCR.[Results]The full length of TpiA gene is 771 bp,encoding 256 amino acid residues in total,and the NCBI accession number is OM906798.According to the deduced amino acid sequence,its molecular weight was predicted to be about 26.97548 kDa,and its isoelectric point was 4.78.The amino acid sequence of the N-terminal signal peptide structure was predicted,and it was found that there was no obvious signal peptide cleavage site,no signal peptide,and no transmembrane region;the amino acid sequence contained 3 N-glycosylation sites,4 protein kinase C phosphorylation sites,2 casein kinase II phosphorylation sites,6 N-myristoylation sites,7 microbody C-terminal target signal site,and 1 triose phosphate isomerase active site.The prediction results of protein subcellular localization showed that TpiA may be located in mitochondria or cytoplasm,with probability of 39.1%and 34.8%,respectively.The amino acid sequence of the TpiA gene of V.alginolyticus shared 98.83%-99.61%homology with other Vibrio species,and it was clustered into the same subfamily with Vibrio parahaemolyticus and had a close relationship.In the secondary structure prediction,the proportions ofα-helix,random coil and extended chain were 44.53%,41.41%and 14.06%,respectively,and the similarity of its tertiary structure model to template 1aw1.1.A was 85.16%.[Conclusions]This study is intended to provide a basis for further research on the role of TpiA gene in the type III secretion system and related research on antibiotic resistance. | Ling ZUO Liangchuan CHEN Shuai YANG Xing XIAO Fuyuan ZENG Junlin WANG Weijie ZHANG Huanying PANG | 2022 | Asian Agricultural Research2022，14，5： | 0 |
| 8 | Cloning and Bioinformatics Analysis of pepck Gene in Vibrio alginolyticus显示文摘[Objectives]To clone the pepck gene of Vibrio alginolyticus strain HY9901 and analyze its sequence by bioinformatics.[Methods]According to the complete gene sequence of V.alginolyticus on GenBank,specific primers were designed to amplify the target gene pepck by PCR.The sequence of the pepck gene was analyzed using bioinformatics.The phylogenic tree of pepck gene and the corresponding single-subunit three-dimensional structure were constructed.[Results]The pepck gene of V.alginolyticus strain HY9901 has a full length of 1629 bp,with theoretical molecular weight of 60.12 kD.The prediction results show that there is no signal peptide or transmembrane region at the N-terminus of the sequence,the amino acid sequence contains 11 phosphorylation sites of casein kinase II.The prediction results of protein subcellular localization indicate that PEPEK protein is localized in the cytoplasm.The protein is stable and hydrophobic.The tertiary structure of the PEPCK protein of V.alginolyticus is similar to that of Vibrio parahaemolyticus.It is predicted that PEPCK has a major functional domain PEPCK_ATP.In the secondary structure,alpha helix,random coil,and extended strand accounted for 21.96%,52.03%and 26.01%,respectively.The PEPCK homology between V.alginolyticus and Vibrio diabolicus is as high as 99%.[Conclusions]This study lays the foundation for further understanding the function of pepck gene in V.alginolyticus. | Fuyuan ZENG Yin ZHAO Shihui ZHOU Chuanhao PAN Miao XIE Huanying PANG | 2020 | Asian Agricultural Research2020，12，8： | 0 |
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