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A 314-bp SINE Insertion in the ZNF2 promoter region may act as a repressor related to regulation of fat deposition in pigs
GU Hao, DU Zhan-yu, Eduard MURANI, Enrico D’ALESSANDRO, CHEN Cai, WANG Xiao-yan, MAO Jiu-de, Klaus WIMMERS, SONG Cheng-yi
2023, 22 (2): 526-536.   DOI: 10.1016/j.jia.2022.08.128
Abstract336)      PDF in ScienceDirect      

Retrotransposons, a type of DNA fragment that can mobilize itself on genome, can generate genetic variations and develop for molecular markers based on the insertion polymorphism.  Zinc finger proteins (ZNFs) are among the most abundant proteins in eukaryotic animals, and their functions are extraordinarily diverse and particularly important in gene regulation.  In the current study, bioinformatic prediction was performed to screen for retrotransposon insertion polymorphisms (RIPs) in six ZNF genes (ZNF2, ZNF3, ZNF7, ZNF8, ZNF10 and ZNF12).  Six RIPs in these ZNFs, including one short interspersed nuclear element (SINE) RIP in intron 1 and one long interspersed nuclear element 1 (L1) RIP in intron 3 of ZNF2, one SINE RIP in 5´ flanking region and one SINE RIP in intron 2 of ZNF3, one SINE RIP in 3´ UTR of ZNF7 and one L1 RIP in intron 2 of ZNF12, were discovered and their presence was confirmed by PCR.  The impact of the SINE RIP in the first intron of ZNF2, which is close to the core promoter of ZNF2, on the gene activity was investigated by dual-luciferase assay in three cell lines.  Our results showed that the SINE insertion in the intron 1 of ZNF2 repressed the core promoter activity extremely significantly (P<0.01) in cervical cancer cells and porcine primary embryonic fibroblasts (HeLa and PEF), thus SINE may act as a repressor.  This SINE RIP also significantly (P<0.05) affected the corrected back fat thickness in Yorkshire pigs.  The corrected back fat thickness of individuals with SINE insertion in the first intron of ZNF2 was significantly (P<0.05) higher than that of individuals without SINE insertion.  In summary, our data suggested that RIPs play important roles in the genetic variations of these ZNF genes and SINE RIP in the intron 1 of ZNF2 may provide a useful molecular marker for the screening of fat deposition in the pig breeding.

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Two new SINE insertion polymorphisms in pig Vertnin (VRTN) gene revealed by comparative genomic alignment
ZHENG Yao, CHEN Cai, CHEN Wei, WANG Xiao-yan, WANG Wei, GAO Bo, Klaus WIMMERS, MAO Jiu-de, SONG Cheng-yi
2020, 19 (10): 2514-2522.   DOI: 10.1016/S2095-3119(20)63255-5
Abstract125)      PDF in ScienceDirect      
Despite one SINE retrotransposon insertion polymorphism (sRTIP) in the vertebrae development-associated (VRTN) gene was identified in pigs, the structural variations (SVs) in VRTN gene and its proximal flank regions were largely unknown.  VRTN genic and flanking sequences from 14 breeds were assembled or downloaded from whole-genome shotgun contings (WGS) database, and aligned to identify the SVs with Clustalx, and retrotransposons in VRTN gene were annotated by RepeatMasker, the splicing patterns of VRTN gene were predicted by Genescan, and large SVs were evaluated by PCR.  A total of 12 small SVs and three large SVs in intron of VRTN, derived from SINE insertion polymorphisms, were identified, and two of them (VRTN-sRTIP2 and VRTN-sRTIP3) were not reported before.  These VRTN-sRTIPs may affect the splicing patterns of VRTN.  They displayed polymorphisms in most detected eight breeds.  VRTN-sRTIP2 and VRTN-sRTIP3 showed Hardy-Weinberg equilibrium distributions in most populations except the Chinese local Erhualian pigs, while VRTN-sRTIP1 showed genetic equilibrium in Erhualian pigs.  Three VRTN-sRTIPs were identified, and displayed polymorphisms in pigs, and two of them were not reported before.  These SVs provide a useful molecular markers for genetic analysis in pigs, and offer new information to facilitate the understanding the SVs of VRTN gene and their putative roles in the variation of vertebral number.
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Pharmacokinetics of oral ethanamizuril solution in chickens
CHENG Pei-pei, HU Xing-xing, WANG Chun-mei, LIU Ying-chun, WANG Mi, ZHANG Ke-yu, FEI Chenzhong, ZHANG Li-fang, WANG Xiao-yang, ZHENG Wen-li, XUE Fei-qun
2018, 17 (12): 2783-2789.   DOI: 10.1016/S2095-3119(18)62056-8
Abstract289)      PDF (895KB)(794)      
Ethanamizuril (EZL) is a novel triazine anticoccidial compound that has high anticoccidial activity in chickens.  In order to treat coccidiosis rationally in poultry, a detection method was developed for ethanamizuril in broiler plasma, and then the pharmacokinetics studies were performed in broilers after oral administration of different dose levels.  Ethanamizuril was administered as single oral doses at low (0.67 mg kg–1 BW), medium (1.33 mg kg–1 BW) and high (6.67 mg kg–1 BW) levels in which the medium dose was that recommended in clinics.  Plasma concentrations of ethanamizuril were determined using ultra-high performance liquid chromatography and the data were analyzed with a non-compartmental model.  Peak plasma concentrations of ethanamizuril were (2.16±0.57), (3.91±0.71), and (23.71±5.02) mg L–1 at (5.17±1.80), (4.60±2.12), and (4.60±2.12) h, respectively.  The terminal elimination half-lives (t1/2λz) for ethanamizuril were (10.84±2.59), (10.66±2.47), and (13.34±3.10) h after oral administration at low, medium and high doses, respectively.  The areas under the concentration-time curve (AUC0–t) were (37.68±6.87), (73.19±9.18), and (485.76±125.10) mg L–1 h with mean residence times (MRT0–t) of (14.79±3.03), (15.57±3.69), and (20.22±4.01) h at the 3 dosages, respectively.  Ethanamizuril was absorbed rapidly and eliminated slowly.  A comparison across the dose range indicated that the time to reach peak plasma concentration (Tmax) values were similar while peak plasma concentration (Cmax) and AUC0–t were positively correlated with increasing dosages.  This study of the pharmacokinetics of an ethanamizuril solution in chickens provides a theoretical basis for the rational use in the clinic.
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Determination of Tetracyclines and Their Epimers in Agricultural Soil Fertilized with Swine Manure by Ultra-High-Performance Liquid Chromatography Tandem Mass Spectrometry
ZHENG Wen-li, ZHANG Li-fang, ZHANG Ke-yu, WANG Xiao-yang, XUE Fei-qun
2012, 12 (7): 1189-1198.   DOI: 10.1016/S1671-2927(00)8646
Abstract1579)      PDF in ScienceDirect      
A rapid, sensitive and specific ultra-high-performance liquid chromatography tandem mass spectrometry (UPLC-MS) method was developed for the analysis of tetracycline antibiotics, including tetracycline (TC), oxytetracycline (OTC), chlortetracycline (CTC) and their 4-epimers (4-epiTCs) in agricultural soil fertilized with swine manure. Soil samples were extracted and cleaned-up with 10 mL EDTA-McIlvaine buffer solution (pH 4.0), then cleaned-up and pre-concentrated using the Oasis MAX cartridge and then eluted with 1 mL solution by mixing formic acid, methanol and water at a ratio of 2:15:83 (v/v/v). The purified samples were separated by an ACQUITY UPLC BEH C18 column using acetonitrile and water containing 0.1% formic acid mobile phase and detected by a single quadrupole MS. The limits of detection for the soil extraction method (LODsoil) ranged from 0.6-2.5 μg kg-1 with recoveries from 23.3-159.2%. Finally, the method was applied to an agricultural field in an area with intensive pig-fattening farming. Tetracyclines were detected in soil from 2.8 to 42.4 μg kg-1 soil. These results demonstrate that soil from swine farms can become severely contaminated with tetracycline antibiotics and their metabolites.
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