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| 1 | Establishment of a transgenic mouse model with liver-specific expression of secretory immunoglobulin D显示文摘Mutation of mevalonate kinase(MVK) is thought to account for most cases of hyperimmunoglobulinemia D syndrome(HIDS) with recurrent fever.However,its mechanism and the relationship between elevated serum immunoglobulin D(IgD) and the clinical features of HIDS are unclear.In this study,we generated by fusion PCR a vector to express high levels of chimeric secretory IgD(csIgD) specifically in the liver.We then generated seven founder lines of transgenic mice by co-microinjection,and verified them using genomic PCR and Southern blotting.We detected the expression of csIgD by reverse transcription PCR,quantitative PCR,western blotting,and enzyme-linked immunosorbent assays.We demonstrated that csIgD could be specifically and stably expressed in the liver.We used flow cytometry to show that overexpression of csIgD in the bone marrow and spleen cells had no effect on B cell development.Morphologic and anatomical observation of the transgenic mice revealed skin damage,hepatosplenomegaly,and nephromegaly in some transgenic mice;in these mice,pathological sections showed high levels of cell necrosis and protein-like sediments in the liver,spleen,and kidney.We demonstrated that the genomic insertion sites of the transgenes did not disrupt the MVK gene on mouse chromosome 5.This transgenic mouse will be useful to explore the pathogenesis of HIDS. | WANG Ping WEI ZhiGuo YAN BoWen HUANG Tan GOU KeMian DAI YunPing ZHENG Min WANG MeiLi CHENG XueQian WANG XiFeng XU Chen SUN Yi | 2012 | Science China(Life Sciences)2012，55，3： | 6 |
| 2 | Pathological features of COVID-19-associated lung injury:a preliminary proteomics report based on clinical samples显示文摘The COVID-19 pandemic has emerged as a global health emergency due to its association with severe pneumonia and relative high mortality.However,the molecular characteristics and pathological features underlying COVID-19 pneumonia remain largely unknown.To characterize molecular mechanisms underlying COVID-19 pathogenesis in the lung tissue using a proteomic approach,fresh lung tissues were obtained from newly deceased patients with COVID-19 pneumonia.After virus inactivation,a quantitative proteomic approach combined with bioinformatics analysis was used to detect proteomic changes in the SARS-CoV-2-infected lung tissues.We identified significant differentially expressed proteins involved in a variety of fundamental biological processes including cellular metabolism,blood coagulation,immune response,angiogenesis,and cell microenvironment regulation.Several inflammatory factors were upregulated,which was possibly caused by the activation of NF-κB signaling.Extensive dysregulation of the lung proteome in response to SARS-CoV-2 infection was discovered.Our results systematically outlined the molecular pathological features in terms of the lung response to SARS-CoV-2 infection,and provided the scientific basis for the therapeutic target that is urgently needed to control the COVID-19 pandemic. | Ling Leng Ruiyuan Cao Jie Ma Danlei Mou Yunping Zhu Wei Li Luye Lv Dunqin Gao Shikun Zhang Feng Gong Lei Zhao Bintao Qiu Haiping Xiang Zhongjie Hu Yingmei Feng Yan Dai Jiang Zhao Zhihong Wu Hongjun Li Wu Zhong | 2020 | Signal Transduction and Targeted Therapy2020，5，1： | 4 |
| 3 | DNA methylation status of H19 and Xist genes in lungs of somatic cell nuclear transfer bovines显示文摘In somatic cell nuclear transfer (SCNT) technologies,the donor cell’s nuclei need to be epigenetically reprogrammed for embryonic development. The incomplete reprogramming of donor cell nuclei has been implicated as a primary reason for the low efficiency of SCNT. DNA methylation is a major epige-netic modification of the genome that regulates crucial aspects of genome function,including estab-lishment of genomic imprinting. In order to make sure whether the DNA methylation reprogramming is efficient in SCNT animals,we analyzed the DNA methylation status of two imprinting genes,H19 and Xist,in lungs of deceased SCNT bovines that died within 48 h of birth using bisulfite sequencing analysis. Our findings demonstrated that cloned bovines showed significantly lower DNA methylation of H19 than controls (P<0.05),and three tested CpGs sites (1,2,3) exhibited unmethylation in one cloned bovine (9C3); however,Xist showed similar DNA methylation levels between clones and con-trols,and both showed hypermethylation (96.11% and 86.67%). | CHEN Jie LI DongJie LIU YanQin ZHANG Cui DAI YunPing LI ShiJie LI Ning | 2008 | Chinese Science Bulletin2008，53，13： | 3 |
| 4 | Production of human lysozyme-transgenic cloned porcine embryos by somatic nuclear transfer显示文摘Due to their physiology and organ size, pigs have significant potential as human disease models and as organ transplantation donors. Genetic modification of pigs could provide benefits for both agriculture and human medicine. In this study, five fetal pig fibroblast cell lines from two species (Wuzhishan and Landrace pigs) were transfected using double-marked human lysozyme (HLY) plasmids (pBC1-HLY-GFP-NEO) by a liposome-mediated method. The ratio of green fluorescent protein (GFP)-expressing cells was >95% in sw7, sw8, slw3 and slw6 cell lines, but only 49.3% in slw9 cells. Cells from the four highly transgenic lines were used as nuclear donors to construct embryos, which were then cultured after fusion and activation by electric stimulation. The rate of cleavage was 76.7%, 48 h after activation. After 7 days, 18.5% of cleaved eggs had developed to the blastocyst stage and 93.3% of blastocysts were GFP-positive. These results indicate that transgenic fetal pig flbroblast cell lines could be obtained by a liposome-mediated method, though the transfection efficiency varied between cell lines. Reconstructed embryos derived from transgenic cells could successfully develop into blastocysts, most of which were GFP-positive. | Qiuyan Li Hengxi Wei Ying Guo Yan Li Rui Zhao Yufang Ma Zhengquan Yu Bo Tang Lei Zhang Yunping Dai Ning Li | 2009 | Progress in Natural Science:Materials International2009，19，6： | 2 |
| 5 | Advances in genetic engineering of domestic animals显示文摘Global population will increase to over nine billion by 2050 with the doubling in demand for meat and milk. To overcome this challenge, it is necessary to breed highly ef ficient and productive livestock. Furthermore,livestock are also excellent models for human diseases and ideal bioreactors to produce pharmaceutical proteins.Thus, genetic engineering of domestic animals presents a critical and valuable tool to address these agricultural and biomedical applications. Overall, genetic engineering has evolved through three stages in history: transgenesis, gene targeting, and gene editing. Since the birth of the first transgenic pig, genetic engineering in livestock has been advancing slowly due to inherent technical limitations. A major breakthrough has been the advent of somatic cell nuclear transfer, which, for the first time, provided the technical ability to produce site-specific genome-modi fied domestic animals. However, the low efficiency of gene targeting events in somatic cells prohibits its wide use in agricultural and biomedical applications. Recently, rapid progress in tools and methods of genome engineering has been made, allowing genetic editing from mutation of a single base pair to the deletion of entire chromosomes.Here, we review the major advances of genetic engineering in domestic animals with emphasis placed on the introduction of latest designer nucleases. | Shaohua WANG Kun ZHANG Yunping DAI | 2016 | Frontiers of Agricultural Science and Engineering2016，3，1： | 1 |
| 6 | Apoptosis of transgenic cloned and recloned bovine blastocysts显示文摘Apoptosis plays an important role in preimplantation embryonic development. Investigating mechanisms of apoptosis can provide useful information for obtaining high-quality embryos and help to improve cloning effciency. Here, we investigated the incidence of blastomere apoptosis in transgenic blastocysts generated by somatic cell nuclear transfer (SCNT) and recloning using a terminal deoxynucleotidyl transferase-mediated d-UTP nick end-labeling (TUNEL) assay. Transgenic recloned embryos were the second generation SCNT embryos derived from the somatic cells of a transgenic SCNT calf. The blastocyst rate of transgenic SCNT embryos was lower than that of nontransgenic SCNT embryos. The incidence of apoptosis in transgenic SCNT embryos was higher than that of nontransgenic SCNT embryos. The blastocyst rate and the incidence of apoptosis in transgenic recloned embryos were similar to nontransgenic SCNT embryos. The process of donor cell transfection and drug selection may decrease the developmental capacity of transgenic SCNT embryos. Serial cloning did not influence the developmental capacity of transgenic recloned embryos. | Guojie Sun Rong Li Yunping Dai Haiping Wang Lili Wang Ying Liu Fangrong Ding Hengxi Wei Ning Li | 2009 | Progress in Natural Science:Materials International2009，19，7： | 1 |
| 7 | Hausdorff Dimensions of Entropy Sets of Dynamical Systems with Positive Entropy显示文摘 | XIONGPING DAI YUNPING JIANG | 2005 | Journal of Statistical Physics2005，，2： | 1 |
| 8 | Effect of leptin on oocyte maturation and subsequent pregnancy rate of cloned embryos reconstructed by somatic cell nuclear transfer in pigs显示文摘由原子转移(SCNT ) 有的体的房间克隆猪在基本研究,人的药和农业生产的宽应用。到改善克隆效率,二基本成熟媒介的效果, NCSU-23 和 TCM199,被比较,并且 TCM199 为有 leptin 的下列实验被选择。我们系统地在 vitro 成熟(IVM ) 在卵母细胞上学习了 leptin 补充的效果,在激活的单性生殖的联盟者(PA ) 和 SCNT 胚胎的 vitro 开发并且在在胚胎转移(et ) 以后的 SCNT 胚胎的 vivo 开发。结果证明进成熟媒介的 100 或 200 ng/ml leptin 的补充极大地没影响 PA 和 SCNT 的卵母细胞,或劈开率的原子成熟(P >
0.05 ) 。当 100 或 200 ng/ml leptin 在 PA 胚囊被加到房间的成熟媒介,和数字时, PA 和 SCNT 胚胎的胚囊率显著地被改进也被改进(P <
0.05 ) 。在 SCNT 的胚囊的房间的数字被改进,当 100 ng/ml leptin 被增加时(P <
0.05 ) 。而且,到 IVM 媒介的 100 或 200 ng/ml leptin 的补充可以在猪克隆改进怀孕率和交货率。 | Hengxi Wei Qiuyan Li Jun Li Yan Li Yunping Dai Yufang Ma Kai Xue Ning Li | 2008 | Progress in Natural Science:Materials International2008，18，12： | 1 |
| 9 | Expression of chromatin modification genes in organs of cloned cattle that died within hours after birth显示文摘Cloning by somatic nuclear transfer is an inefficient process in which many of the cloned ani- mals died shortly after birth and displayed organ ab- normalities. In an effort to determine the possible genetic causes of neonatal death and organ abnor- malities, we have examined expression patterns of four genes that modified chromatin (DNMT1, PCAF,MeCP2 and EED) in six organs (heart, liver, spleen, lung, kidney and brain) of both neonatal death cloned bovines (n=9) and normal control calves produced by artificial insemination (AI) using real-time quantitative RT-PCR. The effect of the age of the fibroblast donor cell on the gene expression profiles was also investigated. Aberrant expressions of DNMT1 and PCAF were found in some studied tissues, but the expression of MeCP2 and EED had similar levels to those of the normal controls. The expression of DNMT1 showed a higher level in heart, liver and brain of both cloned bovines. A higher ex- pression level of PCAF was seen in heart and liver of both cloned bovines, but a lower level was seen only in spleen of adult fibroblast (AF) cell-derived clones. Our results suggest that aberrant expression in gene that modified chromatins were found in cloned bovine tissues of neonatal death. Because DNMT1 and PCAF play an important role in DNA methylation and histone acetylation on nuclear chromatin respectively, and normal expression of DNMT1 and PCAF is needed for precious reprogramming of donor nuclear, the aberrant transcription patterns of DNMT1 andPCAF in these clones may contribute to the defects of organs reported in neonatal death of clones. | LI Shijie LIAN Zhengxing LI Dongjie YU Shuyang ZHANG Lei DAI Yunping LI Rong FEI Jing LI Ning | 2006 | Chinese Science Bulletin2006，51，2： | 1 |
| 10 | Over-expression of CD163, CD169, and CD151 is not sufficient to improve the susceptibility to porcine reproductive and respiratory syndrome virus infection in transgenic mice显示文摘Porcine reproductive and respiratory syndrome virus(PRRSV)is a major pathogen that causes reproductive failure and respiratory disease in pigs,resulting in devastating economic losses worldwide[1].Porcine alveolar macrophages(PAMs)are the primary target cells of PRRSV[2],and the putative receptors,including CD163,CD169,and CD151,play key roles during infection[3–6].However,the understanding of PRRSV infection and pathogenesis is | Zhengzhi Cui Shuaishuai Niu Jingjing Liu Lei Xu Yunping Dai Ning Li Youmin Kang Linlin Zhang Lei Zhou Shuyang Yu | 2017 | Science Bulletin2017，62，24： | 1 |
| 11 | Seasonal influence on the development of parthenotes and cloned embryos in pigs显示文摘Seasonal influence on the developmental ability of porcine embryos was studied by comparisons of the number of cumulus-oocytes-complexes (COCs) per ovary pair and meiotic maturation ability of oocytes in different seasons; and by the observations on developmental competence of the parthenogenetic oocytes and cloned embryos collected from different seasons. We found that the number of COCs per ovary pairs was significantly higher in spring compared to the other seasons (P<0.05). However, no significant difference was found in the rate of oocytes progressing to MⅡ in four seasons (P<0.05). In addition, the rate of blastocyst formation of parthenogenetic embryos in summer was obviously declined compared to the other seasons (P<0.05), and an increased blastocyst rate of cloned porcine embryos was found in spring compared to autumn and winter (P<0.05). The results suggest that there should be a seasonal influence on the developmental competence of porcine embryos. | Zhang Kun Zhang Yunhai Pan Dengke Ma Yufang Wei Hengxi Zhang Jian Li Xuyang Wang Shaohua Li Yan Dai Yunping Li Ning | 2007 | Progress in Natural Science:Materials International2007，17，10： | 0 |
| 12 | A Novel Retrograde AAV Variant for Functional Manipulation of Cortical Projection Neurons in Mice and Monkeys显示文摘Retrograde adeno-associated viruses(AAVs)are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks.However,few retro-grade AAV capsids have been shown to offer access to cor-tical projection neurons across different species and enable the manipulation of neural function in non-human primates(NHPs).Here,we report the development of a novel retro-grade AAV capsid,AAV-DJ8R,which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques.In addition,intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations.Moreover,AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen.These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations. | Yefei Chen Jingyi Wang Jing Liu Jianbang Lin Yunping Lin Jinyao Nie Qi Yue Chunshan Deng Xiaofei Qi Yuantao Li Ji Dai Zhonghua Lu | 2024 | Neuroscience Bulletin2024，40，1： | 0 |
| 13 | Generation of CRISPR/Cas9-mediated lactoferrin-targeted mice by pronuclear injection of plasmid pX330显示文摘Lactoferrin is a member of the transferrin family of multifunctional iron binding glycoproteins.While numerous physiological functions have been described for lactoferrin,the mechanisms underlying these functions are not clear.To further study the functions and mechanisms of lactoferrin,we modified the lactoferrin promoter of mice using the CRISPR/Cas9 system to reduce or eliminate lactoferrin expression.Seven mice with lactoferrin promoter mutations were obtained with an efficiency of 24%(7/29)by injecting the plasmid pX330,expressing a small guide RNA and human codonoptimized SpCas9,into fertilized eggs of mice.Plasmid integration and off-targeting of pX330 were not detected.These results confirmed that pronuclear injection of a circular plasmid is a feasible and efficient method for targeted mutagenesis in mice. | Mengxu GE Fei LIU Fei CHANG Zhaolin SUN Jing FEI Ying GUO Yunping DAI Zhengquan YU Yaofeng ZHAO Ning LI Qingyong MENG | 2015 | Frontiers of Agricultural Science and Engineering2015，2，3： | 0 |
| 14 | Clone of Chinese Jinan red-cross yellow cattle and evaluation of reproductive characteristics of cloned calf显示文摘Somatic cell clone technology is a viable ap- proach to preserving endangered livestock and wildlife ge- netic resources. In the present research, somatic cell nuclear transfer (SCNT) was performed using granulose cells from the critical endangered Chinese red-cross yellow cattle as donor cells. A total of 211 oocytes were manipulated and 166 (79%) of them were successfully enucleated. 112 (67.4%) SCNT embryos were reconstructed, 94 (83%) of them cleaved, and 48 (43%) of them developed to blastocyst stage. SCNT blastocysts were transferred to 6 Holstein recipients, and 2 (33%) of them were found to be pregnant. One of them maintained to term and delivered a calf, whereas another aborted. Effect of different fusion buffer (mannitol vs. Zim- merman fusion buffer) and different activation methods (calcium ionophore+6-DMAP vs. cycloheximide+CB) on fu- sion rate and development of SCNT embryos were investi- gated. The results indicated that: (i) on condition of two DC pulses of 2.5 kV/cm for 10 μs each, fusion rates were higher in mannitol solution than in Zimmerman fusion buffer (71% vs. 61%, respectively, p<0.05), but the blastocysts rates did not differ between two treatments (36% vs. 39%, p>0.05 ); (ii) There was no significant difference in development rates to the blastocyst stage for SCNT embryos activated by calcium ionophore+6-DMAP or by cycloheximide+CB (42% vs. 46%, respectively, p>0.05). Microsatellite DNA analysis examining 28 loci confirmed that the cloned calf was genetically identi- cal to the donor Jinan red-cross yellow cattle and different from the recipient females. Growth and reproductive per- formance of cloned cow were evaluated, and there were no difference i cross-red n it between cloned and normal control Jinan yellow cattle. Furthermore, the cloned yellow cow has delivered a healthy yellow calf. | DAI Yunping LIAN Zhengxing ZHU Huabin GONG Guochun WANG Lili WANG Haiping ZHAO Zhihui ZHU Qinghong FEI Jing LI Ning | 2005 | Chinese Science Bulletin2005，50，22： | 0 |
| 15 | Aberrant histone H4 acetylation in dead somatic cell-cloned calves显示文摘In somatic cell-cloned animals, inefficient epigenetic reprogramming can result in an inappropriate gene expression and histone H4 acetylation is one of the key epigenetic modifications regulating gene expression. In this study, we investigated the levels of histone H4 acetylation of 11 development-related genes and expression levels of 19 genes in lungs of three normal control calves and nine aber- rant somatic cell-cloned calves. The results showed that nine studied genes had decreased acetylation levels in aberrant clones (p < 0.05) and two genes had no significant variations (p > 0.05). Whereas 13 genes had significantly decreased expression (p < 0.05) in aberrant clones, five genes showed no significant differences between controls and clones (p > 0.05), and only one gene had higher expression level in clones (p < 0.05). Furthermore, FGFR, GHR, HGFR and IGF1 genes showed lowered levels of both histone H4 acetylation and expression in aberrant clones than in controls, and the level of histone H4 acetylation was even more lowered in aberrant clones than those in controls. It was suggested that the lower levels of histone H4 acetylation in aberrant clones caused by the previous memory of cell differentiation might not support enough chromatin reprogramming, thus affecting appropriate gene expressions, and growth and development of the cloned calves. To our knowledge, this is the first study on how histone H4 acetylation affects gene expression in organs of somatic cell-cloned calves. | Lei Zhang Shaohua Wang Qiang Li Xiangdong Ding Yunping Dai Ning Li | 2008 | Progress in Natural Science:Materials International2008，18，7： | 0 |
| 16 | Expression of recombinant human butyrylcholinesterase in the milk of transgenic mice显示文摘Butyrylcholinesterase(BCHE)is a natural bioscavenger that protects humans against organophosphate toxicity.Due to the limited yield of human BCHE(hBCHE)when purifying from human plasma,it is necessary to find an alternative method to produce this protein.One potential method is to produce transgenic livestock that make modified milk containing high concentration of hBCHE.In this study,we cloned the hBCHEgene into a human lactoferrin(hLF)bacterial artificial chromosome(BAC)construct to make a hLFhBCHE BAC construct.Subsequently,we injected the BAC construct into pronuclei of mouse fertilized embryos and generated transgenic mice.Expression analysis showed that recombinant hBCHE(rhBCHE)was expressed efficiently in the mammary gland of the transgenic mice and the concentration of rhBCHE in the milk of individual mice ranged from 7612 to 15928 mg·L^(–1).Protein function tests showed that rhBCHE has the same enzymatic activity as the native hBCHE.Our results pave the way for making transgenic livestock to produce large quantities of rhBCHE. | Dan LU Shengzhe SHANG Shen LIU Ying WU Fangfang WU Tan TAN Qiuyan LI Yunping DAI Xiaoxiang HU Yaofeng ZHAO Ning LI | 2014 | Frontiers of Agricultural Science and Engineering2014，1，3： | 0 |
| 17 | Depletion of conventional mature B cells and compromised specific antibody response in bovine immunoglobulin μ heavy-chain transgenic mice显示文摘In this study,we introduced the bovine immunoglobulinμheavy-chain gene(the orphaned gene on BTA11)into mouse germline cells.Bovine IgM was highly expressed in selected transgenic lines,and it largely inhibited rearrangements of the endogenous immunoglobulin heavy chain(IgH)genes in these lines.The forced expression of bovine IgM resulted in reduced numbers of pro-and pre-B cells but increased the number of immature B cells in the transgenic mice.Bovine IgM-expressing B cells can migrate from the bone marrow to the spleen,but most of the cells are arrested at the T1 transitional B cell stage,leading to a significantly lower number of T2 transitional and mature B cells in the spleen.Although the serum concentrations of endogenous IgM and IgG in the transgenic mice were significantly decreased,the IgA levels were slightly increased compared to the WT mice.The bovine IgM level in the serum was only one-tenth to one-fifth of that of endogenous mouse IgM,suggesting that most of the serum immunoglobulin were contributed by endogenous IgH gene-expressing B cells.These transgenic mice also exhibited a lower frequency of unique complementarity determining region 3(CDR3)sequences in their VH repertoire and Vκrepertoire but exhibited an increased frequency of unique CDR3 in their Vλrepertoire.Compared to the WT mice,the transgenic mice had a significantly higher percentage of mouse IgMexpressing B cells that expressedλchains.Finally,we showed that the transgenic mice were deficient in a specific antibody response to antigen stimulation. | Min ZHANG Xueqian CHENG Dan CHU Jingwen LIANG Yi SUN Li MA Beilei XU Min ZHENG Meili WANG Liming REN Xiaoxiang HU Qingyong MENG Ran ZHANG Ying GUO Yunping DAI Robert AITKEN Ning LI Yaofeng ZHAO | 2014 | Frontiers of Agricultural Science and Engineering2014，1，2： | 0 |
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