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促黄体激素（LH） 1

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半滑舌鳎 1

卵母细胞去核 1

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实时荧光定量PCR 1

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Effects of enucleation method on

Chengcheng ZHAO, Junsong SHI, Rong ZHOU, Ranbiao MAI, Lvhua LUO, Xiaoyan HE, Hongmei JI, Gengyuan CAI, Dewu LIU, Enqin ZHENG, Zhenfang WU, Zicong LI

《农业科学与工程前沿（英文）》 2019年 第6卷 第1期   页码 61-65 doi: 10.15302/J-FASE-2018227

摘要：

Enucleation is a crucial procedure for mammalian somatic cell nuclear transfer (SCNT), especially for domestic animal cloning. Oocytes of domestic animals such as pigs and cattle contain dark lipid droplets that hinder localization and removal of the nucleus. Using an oocyte enucleation technique that can obtain a high enucleation rate but has minimal negative effects on the reprogramming potential of oocyte for cloning is beneficial for enhancing the outcome of SCNT. In this study, we compared the pig cloning efficiency resulting from blind aspiration-based (BA-B) enucleation and spindle imaging system-assisted (SIS-A) enucleation, and compared the pig SCNT success rate associated with BA-B enucleation and blind aspiration plus post-enucleation staining-based (BAPPS-B) enucleation. SIS-A enucleation achieved a significantly higher oocyte enucleation success rate and tended to obtain a higher full term development rate of SCNT embryos than BA-B enucleation. BAPPS-B enucleation also obtained significantly higher as well as full term development efficiency of cloned porcine embryos than BA-B enucleation. These data indicate that SIS-A and BAPPS-B enucleation are better approaches for pig SCNT than BA-B enucleation.

关键词： cloning     enucleation     pig     SCNT    

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EFFICIENT CONTAMINANT REMOVAL FROM LIQUID DIGESTATE OF PIG MANURE BY CHEMICAL PRECIPITATION AND CO MINERALIZATION

《农业科学与工程前沿（英文）》 2023年 第10卷 第3期   页码 479-491 doi: 10.15302/J-FASE-2023480

摘要：

● LFD was treated by fly ash-based chemical precipitation and CO2 mineralization.

关键词： anaerobic digestion     chemical oxygen demand     fly ash     ion removal     total phosphate    

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EFFECT OF EXOGENOUS ADDITIVES ON HEAVY METAL PASSIVATION AND NITROGEN RETENTION IN PIG MANURE COMPOSTING

《农业科学与工程前沿（英文）》 doi: 10.15302/J-FASE-2023487

摘要：

● Research on heavy metal passivation and nitrogen emissions is necessary for the pig industry.

关键词： additives     composting     heavy metals passivation     nitrogen retention     pig manure    

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Factors influencing the somatic cell nuclear transfer efficiency in pigs

Yong JIN, Manling ZHANG, Xinrong JU, Shuang LIANG, Qiang XIONG, Lihua ZHAO, Xiaowei NIE, Daorong HOU, Qiang LIU, Junzheng WANG, Chenyu WANG, Xiaokang LI, Lining ZHANG, Xiaorui LIU, Ying WANG, Haiyuan YANG, Yifan DAI, Rongfeng LI

《农业科学与工程前沿（英文）》 2019年 第6卷 第1期   页码 73-80 doi: 10.15302/J-FASE-2018220

摘要：

Using a data set from our laboratory, we assessed the effects of several factors on pig cloning efficiency. The results demonstrated that cells at high confluence (>90%) used as donor cell resulted in higher pregnancy rate, delivery rate and overall cloning efficiency (number of live offspring born per reconstructed embryo transferred to recipients) compared with the cells at 60% to 79% confluence and 80% to 89% confluence. Cells with four, five and six passages compromised the pregnancy and delivery rates compared with first passage cells. The number of blastocysts transferred by somatic cell nuclear transfer (SCNT) did not significantly affect the cloning efficiency, but transfer of blastocyst derived from culture 5 d after SCNT achieved a significantly higher pregnancy rate compared with one to two cell SCNT embryos from overnight culture. The highest pregnancy rate, delivery rate and the largest litter size were obtained when Bama Miniature pig fibroblasts were used as donor cells and Landrace/Yorkshire hybrid gilts were used as recipients. Recipients treated with chemicals for estrus synchronization had higher pregnancy rates compared with untreated recipients. Our data might be helpful for improving SCNT efficiency in pigs.

关键词： blastocyst     donor cell     estrus synchronization     pregnancy rate     pig cloning     somatic cell nuclear transfer    

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AMMONIA DISPERSION FROM MULTI-FLOOR VERSUS STANDARD SINGLE-FLOOR PIG PRODUCTION FACILITIES BASED ON COMPUTATIONAL

《农业科学与工程前沿（英文）》 2023年 第10卷 第3期   页码 374-389 doi: 10.15302/J-FASE-2023501

摘要：

● NH3 dispersion from a multi-floor pig building was compared to a single-floor building.

关键词： pig building     computational fluid dynamics     ammonia     dispersion    

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Measurement of trimethylamine concentration and evaluation of pig meat natural quality by a spectrophotometric

Mohammod Abdul HAMID,Xi WANG,Xiangdong DING,Chuduan WANG,Xingbo ZHAO

《农业科学与工程前沿（英文）》 2014年 第1卷 第3期   页码 236-241 doi: 10.15302/J-FASE-2014034

摘要： Pig meat off-flavor is attributed to trimethylamine (TMA) concentration, and it is considered as the precursor of the fishy off-flavor problems. In this study, TMA concentrations in pig meat were determined, and the interactions with breed and gender effects were discussed. In addition, the TMA threshold for meat off-flavor and pig meat natural quality was measured in relation to meat storage and movement, and the influential factors including the pig breed and storage time were discussed. The results indicated positive effects on the precursor of the fishy off-flavor and the TMA threshold. Native breeds were found to have lower TMA concentrations than European breeds ( <0.01), and females and castrated males had significantly lower TMA concentration than males ( <0.01), The threshold concentration of TMA when meat was classed as off-flavored was 25 μg·g , and this occurred after 35–38 h of storage. The natural qualities, such as appearance, flavor, color and overall acceptable scores declined significantly after 4 days in storage ( <0.01). It is concluded that pig meat off-flavor, breed and gender were essential factors affecting flavor for meat breeding programs, and storage time is important for pig meat natural quality.

关键词： pig meat     trimethylamine (TMA)     influential factor    

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Reprogramming of the pig primordial germ cells into pluripotent stem cells: a brief review

Qijing LEI, Qin PAN, Shuai YU, Na LI, Shulin CHEN, Kuldip SIDHU, Jinlian HUA

《农业科学与工程前沿（英文）》 2019年 第6卷 第1期   页码 28-32 doi: 10.15302/J-FASE-2018222

摘要：

Primordial germ cells (PGCs) are regarded as unipotent cells that can produce only either spermatogonia or oocytes. However, PGCs can be converted into the pluripotent state by first dedifferentiation to embryonic germ cells and then by reprogramming to induce them to become pluripotent stem cells (iPSCs). These two stages can be completely implemented with mouse cells. However, authentic porcine iPSCs have not been established. Here, we discuss recent advances in the stem cell field for obtaining iPSCs from PGCs. This knowledge will provide some clues which will contribute to the regulation of reprogramming to pluripotency in farm species.

关键词： pig     pluripotent stem cells     primordial germ cells     reprogramming    

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人类遗传病的家系收集疾病基因定位克隆与疾病基因功能的研究

夏家辉

《中国工程科学》 2000年 第2卷 第11期   页码 1-11

摘要：

介绍了中国医学遗传学国家重点实验室在遗传病家系收集、疾病基因定位、疾病基因克隆和疾病基因功能研究方面的研究工作。用细胞遗传学G显带技术于1975年发现了一条与鼻咽癌相关的标记染色体t（1；3）（q44；p11）；1981年将睾丸决定基因（TDF）定位于Yp11.32带；1991年以来收集遗传病家系345种共590个；1996年用显微切割、PCR、微克隆技术克隆了EXT2基因；1998年用基因家族-候选疾病基因克隆方法克隆了遗传性神经性耳聋基因GJB3；1999年用连锁分析和全基因组扫描将一种遗传性弥漫性浅表性光敏性汗孔角化症定位于12q23.2带，并在基因功能研究中发现了一个新的细胞内转运蛋白。

关键词： 遗传病家系     基因定位和克隆     基因家族-候选疾病基因克隆     基因组扫描     基因功能研究    

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Genome-wide association study of the backfat thickness trait in two pig populations

Dandan ZHU,Xiaolei LIU,Rothschild MAX,Zhiwu ZHANG,Shuhong ZHAO,Bin FAN

《农业科学与工程前沿（英文）》 2014年 第1卷 第2期   页码 91-95 doi: 10.15302/J-FASE-2014005

摘要： Backfat thickness is a good predictor of carcass lean content, an economically important trait, and a main breeding target in pig improvement. In this study, the candidate genes and genomic regions associated with the tenth rib backfat thickness trait were identified in two independent pig populations, using a genome-wide association study of porcine 60K SNP genotype data applying the compressed mixed linear model (CMLM) statistical method. For each population, 30 most significant single-nucleotide polymorphisms (SNPs) were selected and SNP annotation implemented using Build 10.2. In the first population, 25 significant SNPs were distributed on seven chromosomes, and SNPs on SSC1 and SSC7 showed great significance for fat deposition. The most significant SNP (ALGA0006623) was located on SSC1, upstream of the gene. In the second population, 27 significant SNPs were recognized by annotation, and 12 SNPs on SSC12 were related to fat deposition. Two haplotype blocks, M1GA0016251-MARC0075799 and ALGA0065251-MARC0014203-M1GA0016298-ALGA0065308, were detected in significant regions where the and genes were identified as contributing to fat metabolism. The results indicated that genetic mechanism regulating backfat thickness is complex, and that genome-wide associations can be affected by populations with different genetic backgrounds.

关键词： backfat thickness     SNP chip     genome-wide association study     compressed mixed linear model     pig    

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Detection of genomic signatures for pig hairlessness using high-density SNP data

Ying SU,Yi LONG,Xinjun LIAO,Huashui AI,Zhiyan ZHANG,Bin YANG,Shijun XIAO,Jianhong TANG,Wenshui XIN,Lusheng HUANG,Jun REN,Nengshui DING

《农业科学与工程前沿（英文）》 2014年 第1卷 第4期   页码 307-313 doi: 10.15302/J-FASE-2014039

摘要： Hair provides thermal regulation for mammals and protects the skin from wounds, bites and ultraviolet (UV) radiation, and is important in adaptation to volatile environments. Pigs in nature are divided into hairy and hairless, which provide a good model for deciphering the molecular mechanisms of hairlessness. We conducted a genomic scan for genetically differentiated regions between hairy and hairless pigs using 60K SNP data, with the aim to better understand the genetic basis for the hairless phenotype in pigs. A total of 38405 SNPs in 498 animals from 36 diverse breeds were used to detect genomic signatures for pig hairlessness by estimating between-population ( ) values. Seven diversifying signatures between Yucatan hairless pig and hairy pigs were identified on pig chromosomes (SSC) 1, 3, 7, 8, 10, 11 and 16, and the biological functions of two notable genes, and were revealed. When Mexican hairless pigs were contrasted with hairypigs, strong signatures were detected on SSC1 and SSC10, which harbor two functionally plausible genes, and . KEGG pathway analysis showed a subset of overrepresented genes involved in the T cell receptor signaling pathway, MAPK signaling pathway and the tight junction pathways. All of these pathways may be important in local adaptability of hairless pigs. The potential mechanisms underlying the hairless phenotype in pigs are reported for the first time. and are interesting candidate genes for the hairless phenotype in Yucatan hairless and Mexico hairless pigs, respectively. , , and as well as other genes involved in the MAPK and T cell receptor signaling pathways may be important in environmental adaption by improved tolerance to UV damage in hairless pigs. These findings improve our understanding of the genetic basis for inherited hairlessness in pigs.

关键词： hairlessness     pig     selective sweeping    

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Factors affecting early embryonic development in cattle: relevance for bovine cloning

Yanna DANG, Kun ZHANG

《农业科学与工程前沿（英文）》 2019年 第6卷 第1期   页码 33-41 doi: 10.15302/J-FASE-2018228

摘要：

Female infertility represents a major challenge for improving the production efficiency in the dairy industry. Historically, fertility has declined whereas milk yield has increased tremendously due to intensive genetic selection. evidence reveals about 60% pregnancy loss takes place during the first month following fertilization. Meanwhile, early embryo development is significant for somatic cell nuclear transfer in cattle as a large proportion of cloned embryos fail to develop beyond peri-implantation stage. Oocyte quality is of utmost importance for the early embryo to develop to term for both fertilized and cloned embryos. Epigenetic reprogramming is a key process occurring after fertilization and critical roles of epigenetic modifiers during preimplantation development are now clear. Incomplete epigenetic reprogramming is believed to be a major limitation to cloning efficiency. Treatment of cloned embryos with epigenetic modifying drugs (e.g., Trichostatin A) could greatly improve cloning efficiency in both mice and cattle. Recently, the rapid progress in high-throughput sequencing technologies has enabled detailed deciphering of the molecular mechanisms underlying these events. The robust efficiency of genomic editing tools also presents an alternative approach to the functional annotation of genes critical to early development.

关键词： bovine cloning     embryo development     somatic cell nuclear transfer     X-inactive specific transcript    

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Validation of

Youfen Xu, Zong Li, Ruyin Liu, Hongxia Liang, Zhisheng Yu, Hongxun Zhang

《环境科学与工程前沿（英文）》 2020年 第14卷 第4期 doi: 10.1007/s11783-020-1246-9

摘要： Abstract • Pig feces is the predominant excrement produced by animal husbandry in China. • The PF, Pig-1-BacTaqMan, and Pig-2-BacTaqMan MST assays showed better performance. • The pig-specific MST assays can contribute to managing the pig fecal pollution. In China, pig feces is the predominant source of excrement produced by animal husbandry. Improper use or direct discharge of pig feces can result in contamination of natural water systems. Microbial source tracking (MST) technology can identify the sources of fecal pollution in environmental water, and contribute to the management of pig fecal pollution by local environmental protection agencies. However, the accuracy of such assays can be context-dependent, and they have not been comprehensively evaluated under Chinese conditions. We aimed to compare the performance of five previously reported pig-specific MST assays (PF, Pig-Bac1SYBR, Pig-Bac2SYBR, Pig-1-BacTaqMan, and Pig-2-BacTaqMan, which are based on Bacteroidales 16S rRNA gene markers) and apply them in two rivers of North China. We collected a total of 173 fecal samples from pigs, cows, goats, chickens, humans, and horses across China. The PF assay optimized in this study showed outstanding qualitative performance and achieved 100% specificity and sensitivity. However, the two SYBR green qPCR assays (Pig-Bac1SYBR and Pig-Bac2SYBR) cross-reacted with most non-pig fecal samples. In contrast, both the Pig-1-BacTaqMan and Pig-2-BacTaqMan assays gave 100% specificity and sensitivity. Of these, the Pig-2-BacTaqMan assay showed higher reproducibility. Our results regarding the specificity of these pig-specific MST assays differ from those reported in Thailand, Japan, and America. Using the PF and Pig-2-BacTaqMan assays, a field test comparing the levels of pig fecal pollution in rivers near a pig farm before and after comprehensive environmental pollution governance indicated that pig fecal pollution was effectively controlled at this location.

关键词： Microbial source tracking     Pig fecal pollution     16S rRNA gene markers     Pig-specific Bacteroidales    

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半滑舌鳎促黄体激素基因克隆和表达分析及其血清浓度测定

柳学周,史宝,王珊珊,徐永江,李晓晓

《中国工程科学》 2014年 第16卷 第9期   页码 50-60

摘要：

利用RACE技术首次克隆了半滑舌鳎脑垂体中促黄体激素（LH）基因cDNA全长序列。该基因全长670 bp，开放阅读框为477 bp，编码158 个氨基酸。与其他脊椎动物的LH成熟肽氨基酸序列同源性比较表明：半滑舌鳎LH与鲽形目和鲈形目同源性高58 %～68 %。另外，半滑舌鳎LH含有12 个保守的半胱氨酸残基和1 个N-糖基化位点（19～21 NQT）。实时荧光定量组织表达分析表明，LH mRNA在脑、垂体、卵巢等组织中表达，垂体中表达量最丰富，其他组织尤以脑、性腺表达量较高，推测半滑舌鳎LH可能具有广泛的垂体外生理功能；实时荧光定量PCR方法对繁殖周期雌性半滑舌鳎LH表达水平进行测定，结果表明，LH mRNA在Ⅱ～Ⅵ各繁殖周期的脑、垂体、卵巢3 种组织都有表达，但表达水平有差异，在Ⅳ、Ⅴ期表达量最丰富，说明LH主要促进卵母细胞最终成熟及排卵。利用125I 标记的放射免疫测定（RIA）技术检测繁殖周期半滑舌鳎血清LH浓度，结果表明LH在Ⅴ期含量最丰富，也说明LH主要促进卵母细胞最终成熟及排卵。

关键词： 半滑舌鳎     促黄体激素（LH）     cDNA克隆     实时荧光定量PCR     放射免疫测定    

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全球首例猪心脏移植人体手术

Sarah C. P. Williams

《工程（英文）》 2022年 第16卷 第9期   页码 6-8 doi: 10.1016/j.eng.2022.07.006

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基于细胞内应变评估的卵母细胞去核方法及其在机器人化克隆中的应用 Article

孙明竹, 刘曜玮, 崔茂盛, 赵启立, 赵相飞, 张艺荻, 黄晶晶, 卢桂章, 赵新

《工程（英文）》 2023年 第24卷 第5期   页码 73-83 doi: 10.1016/j.eng.2022.04.016

摘要：

自1996 年第一只克隆羊诞生，克隆技术由于在动物育种方面的巨大潜力而得到了很大关注。体细胞核移植技术是动物克隆的核心，其操作过程非常复杂，不可避免地造成细胞内损伤。通常来说，只有不到1%的重构胚胎能够发育成克隆动物，克隆的低成功率是制约其广泛应用的主要原因之一。为此，本文提
出了一种基于细胞内应变评估的卵母细胞去核方法，以减少体细胞核移植中的潜在细胞内损伤，从而提高克隆成功率。首先，根据细胞内速度场计算细胞内应变，进而将内应变作为评价标准来改进去核操作；在此基础上，开发了一套机器人批量体细胞核移植系统，用于实现机器人化克隆。实验结果表明，本文方法将体细胞核移植后的胚胎囊胚率从10.0%提高到20.8%，并成功获得17 头克隆小猪。这是世界上首批由机器人完成克隆操作获得的克隆动物。对比手动核移植操作，本文将克隆成功率从平均0.73%提升到了2.50%，大幅提升了克隆效率。与此同时，基于细胞内应变的去核方法有望应用于其他生物操作，并建立一种通用的减少细胞内损伤的细胞操作规范。

关键词： 卵母细胞去核     机器人化克隆     细胞内应变     细胞内损伤     光流法    

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标题 作者 时间 类型 操作

Effects of enucleation method on

Chengcheng ZHAO, Junsong SHI, Rong ZHOU, Ranbiao MAI, Lvhua LUO, Xiaoyan HE, Hongmei JI, Gengyuan CAI, Dewu LIU, Enqin ZHENG, Zhenfang WU, Zicong LI

期刊论文

EFFICIENT CONTAMINANT REMOVAL FROM LIQUID DIGESTATE OF PIG MANURE BY CHEMICAL PRECIPITATION AND CO MINERALIZATION

期刊论文

EFFECT OF EXOGENOUS ADDITIVES ON HEAVY METAL PASSIVATION AND NITROGEN RETENTION IN PIG MANURE COMPOSTING

期刊论文

Factors influencing the somatic cell nuclear transfer efficiency in pigs

Yong JIN, Manling ZHANG, Xinrong JU, Shuang LIANG, Qiang XIONG, Lihua ZHAO, Xiaowei NIE, Daorong HOU, Qiang LIU, Junzheng WANG, Chenyu WANG, Xiaokang LI, Lining ZHANG, Xiaorui LIU, Ying WANG, Haiyuan YANG, Yifan DAI, Rongfeng LI

期刊论文

AMMONIA DISPERSION FROM MULTI-FLOOR VERSUS STANDARD SINGLE-FLOOR PIG PRODUCTION FACILITIES BASED ON COMPUTATIONAL

期刊论文

Measurement of trimethylamine concentration and evaluation of pig meat natural quality by a spectrophotometric

Mohammod Abdul HAMID,Xi WANG,Xiangdong DING,Chuduan WANG,Xingbo ZHAO

期刊论文

Reprogramming of the pig primordial germ cells into pluripotent stem cells: a brief review

Qijing LEI, Qin PAN, Shuai YU, Na LI, Shulin CHEN, Kuldip SIDHU, Jinlian HUA

期刊论文

人类遗传病的家系收集疾病基因定位克隆与疾病基因功能的研究

夏家辉

期刊论文

Genome-wide association study of the backfat thickness trait in two pig populations

Dandan ZHU,Xiaolei LIU,Rothschild MAX,Zhiwu ZHANG,Shuhong ZHAO,Bin FAN

期刊论文

Detection of genomic signatures for pig hairlessness using high-density SNP data

Ying SU,Yi LONG,Xinjun LIAO,Huashui AI,Zhiyan ZHANG,Bin YANG,Shijun XIAO,Jianhong TANG,Wenshui XIN,Lusheng HUANG,Jun REN,Nengshui DING

期刊论文

Factors affecting early embryonic development in cattle: relevance for bovine cloning

Yanna DANG, Kun ZHANG

期刊论文

Validation of

Youfen Xu, Zong Li, Ruyin Liu, Hongxia Liang, Zhisheng Yu, Hongxun Zhang

期刊论文

半滑舌鳎促黄体激素基因克隆和表达分析及其血清浓度测定

柳学周,史宝,王珊珊,徐永江,李晓晓

期刊论文

全球首例猪心脏移植人体手术

Sarah C. P. Williams

期刊论文

基于细胞内应变评估的卵母细胞去核方法及其在机器人化克隆中的应用

孙明竹, 刘曜玮, 崔茂盛, 赵启立, 赵相飞, 张艺荻, 黄晶晶, 卢桂章, 赵新

期刊论文