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Production of Transgenic Mice by Type-A Spermatogonia-Mediated Gene Transfer

Agricultural Sciences in China 2011年第3期10卷 431-437页

作者： ju hui-ming BAI Li-jing REN Hong-yan MU Yu-lian YANG Shu-lin LI Kui Key Laboratory for Farm Animal Genetic Resources and Utilization Ministry of Agriculture~Institute of Animal Science Chinese Academyof Agricultural Sciences Beijing 100193 P.R. China College of Veterinary Medicine Yangzhou University Yangzhou 225009 P.R. China

Type-A spermatogonia first appear at between 3-7 d postnatally in mice and are the only immortalized diploid cells that reproduce in adulthood in these animals. In our current study, we explored the feasibility of producing stable transgenic mice using these cells. Enhanced pEGFP-N1 plasmids were suspended in ExGen500 transfection reagent and injected at different angles into the testes of 7-d-old male ICR mice. The resulting type-A spermatogonia-mediated gene transfer （TASMGT） mice were then mated with normal females at different stages of sexual maturity （6, 12, and 24 wk）. The integration and expression of the introduced EGFP gene was evaluated in the F1 transgenic offspring by PCR and Southern blotting analysis. The foreign gene integration rates for a low-dose group （15 μL gene suspension injected into each testis） and a high-dose group （30 μL suspensions injected） at the three stages of female sexual maturity tested were 11.76% （2/17）, 14.29% （3/21）, and 11.11% （2/18）, and 5% （1/20）, 5.56% （1/18）, and 0 （0/17）, respectively. The average integration rates for these two dose groups were 12.5% （7/56） and 3.64% （2/55）, respectively, which was a significant difference （P0.05）. Semi-quantitative RT-PCR analysis further showed that the introduced GFP gene was expressed in 3/9 integration mice. In addition, GFP expression was observed in the sperm cells from the TASMGT mice, and also in the embryos and F2 pups from the F1 generation transgenic mice. Hence, although the foreign gene integration rate for TASMGT is not high and the transgenic offspring show as yet unexplained defects, our results indicate that this method is a potentially feasible and reproducible new approach to creating transgenic mice.

关键词： type-A spermatogonia transgenic mice enhanced green fluorescent protein type-A spermatogonia-mediated gene transfer

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The Establishment of Double-Transgenic Mice that Co-Express the appA and MxA Genes Mediated by Type A Spermatogonia In vivo

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Journal of Integrative Agriculture 2014年第12期13卷 2741-2749页

作者： BAI Li-jing ju hui-ming MU Yu-lian YANG Shu-lin REN Hong-yan AO Hong WANG Chu-duan LI Kui State Key Laboratory for Animal Nutrition Ministry of Agriculture/Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation Ministry of Agriculture/Institute of Animal Science Chinese Academy of Agricultural Sciences College of Animal Science China Agricultural University College of Veterinary Medicine Yangzhou University

Type A spermatogonial stem cells are the only immortal diploid cells in the postnatal animal that undergo self-renewal through the lifetime of an animal and transmit genes to subsequent generations. In this paper, the generation and characterization of double-transgenic mice co-expressing the Escherichia coli appA gene and human MxA gene generated via the in vivo transfection of type A spermatogonial cells were reported for the ifrst time. The dicistronic expression vector pcDNA-appA-MxA（AMP） and ExGen500 transfection reagent were injected into the testicular tissue of 7-d-old male ICR mice. The mice that underwent testis-mediated gene transfer were mated with wild-type female mice, and the integration and expression of the foreign genes in the offspring were evaluated. Transgenic mice that co-expressed appA and MxA showed a gene integration rate of 8.89%（16/180）. The transgenic mice were environmentally friendly, as the amount of phosphorous remaining in the manure was reduced by as much as 11.1%by the appA gene （P〈0.05）;these animals also exhibited a strong anti-viral phenotype.

关键词： type A spermatogonial stem cells dicistronic expression vector co-expression transgenic mice

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GPS: graph contrastive learning via multi-scale augmented views from adversarial pooling

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Science China(Information Sciences) 2025年第1期68卷 145-158页

作者： Wei ju Yiyang GU Zhengyang MAO Ziyue QIAO Yifang QIN Xiao LUO hui XIONG ming ZHANG School of Computer Science National Key Laboratory for Multimedia Information ProcessingPeking University Artificial Intelligence Thrust The Hong Kong University of Science and Technology Department of Computer Science University of California

Self-supervised graph representation learning has recently shown considerable promise in a range of fields, including bioinformatics and social networks. A large number of graph contrastive learning approaches have shown promising performance for representation learning on graphs, which train models by maximizing agreement between original graphs and their augmented views(i.e., positive views). Unfortunately, these methods usually involve pre-defined augmentation strategies based on the knowledge of human experts. Moreover, these strategies may fail to generate challenging positive views to provide sufficient supervision signals. In this paper, we present a novel approach named graph pooling contrast(GPS) to address these *** by the fact that graph pooling can adaptively coarsen the graph with the removal of redundancy, we rethink graph pooling and leverage it to automatically generate multi-scale positive views with varying emphasis on providing challenging positives and preserving semantics, i.e., strongly-augmented view and weakly-augmented view. Then, we incorporate both views into a joint contrastive learning framework with similarity learning and consistency learning, where our pooling module is adversarially trained with respect to the encoder for adversarial robustness. Experiments on twelve datasets on both graph classification and transfer learning tasks verify the superiority of the proposed method over its counterparts.

关键词： graph representation learning graph neural networks graph contrastive learning graph augmentations graph pooling

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Bufalin Suppresses Esophageal Squamous Cell Carcinoma Progression through Protein Arginine Methyltransferase-6/AKT/Mammalian Target of Rapamycin Pathway Inhibition

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World Journal of Traditional Chinese Medicine 2025年第01期 148-156页

作者： Yao-Ke Jia Qian-Qian ju hui Shi Xin-Xin Si Department of Clinical Medicine Nanjing Medical University Department of Clinical Medicine Medical College of Nantong University Department of Thoracic Surgery School of Medicine Shanghai Chest Hospital Shanghai Jiao Tong University Department of Medical Biology School of Pharmacy Jiangsu Ocean University

Objective: Esophageal squamous cell carcinoma(ESCC), a commonly encountered malignant tumor in the gastrointestinal tract, poses a significant health burden. Bufalin, a pharmacologically active molecule, has been shown to exhibit antitumor activity against various types of cancers. This study investigates the molecular mechanism underpinning the effects of bufalin on ESCCs. Materials and Methods: The impact of bufalin on the proliferation and migration of ESCC cells was evaluated through the utilization of the Cell Counting Kit-8(CCK-8), scratch assay, and transwell assay. In addition, ribonucleic acid(RNA) sequencing was performed to identify genes that were abnormally expressed in response to bufalin treatment. Western blotting was utilized to ascertain the expression levels of protein arginine methyltransferase-6(PRMT6), phosphorylated AKT(p-AKT), and phosphorylated mammalian target of rapamycin(p-m TOR). Cell transfection was then performed to observe the rescue effect of PRMT6 on bufalin. Results: Bufalin displayed a significant time-dependent inhibition of the proliferation, migration, and invasion of ECA109 cells. An RNA sequencing(RNA-seq) analysis revealed that PRMT6 expression was downregulated in the cells treated with bufalin. PRMT6 promoted the proliferation, migration, and invasive potential of ECA109 cells. The overexpression of PRMT6 boosted p-AKT and p-m TOR levels in ECA109 cells and reversed bufalin inhibition. Conclusions: These findings indicate that bufalin exerts its inhibitory effects on ESCCs through the PRMT6/AKT/m TOR signaling pathway. These findings lay the groundwork for bufalin as a promising therapeutic candidate for the treatment of ESCC.

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整合型诱导表达猪生长激素(pGH)载体的构建及表达研究

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畜牧兽医学报 2013年第7期44卷 1008-1013页

作者：鞠辉明白立景姜星李奎扬州大学兽医学院扬州225009 中国农业科学院北京畜牧兽医研究所/农业部畜禽遗传资源与利用重点开放实验室北京100193

旨在构建整合型诱导表达猪生长激素(Porcine growth hormone,pGH)载体,在细胞水平验证其诱导效率及表达效率。从pTRE-GH12载体上扩增TRE-GH片段,通过Sal I酶切位点连接到pCAGGS-rtTA载体中,构建重组载体pCAGGS-rtTA-TRE-GH12(下称pTTGH),将pTTGH质粒转染猪PK15细胞,经G418筛选后,在培养液中添加诱导底物强力霉素(Doxycycline,DOX)后不同时间段通过实时荧光定量PCR(QRT-PCR)测定细胞内pGH mRNA的表达;在培养液中添加不同浓度的的DOX,通过QRT-PCR及Western blot测定细胞内pGH的表达变化。酶切及测序结果表明成功构建了pTTGH载体;QRT-PCR结果表明,和对照组相比,试验组细胞培养液中添加诱导底物后外源GH基因表达在36h时最高;在一定浓度范围内,外源GH表达水平和添加DOX的浓度呈现出正相关性,而正常细胞组中GH的表达水平不受DOX添加与否及量的影响。通过条带灰度分析Western杂交结果验证了培养基中一定范围内DOX浓度和pGH表达量呈现出正相关。试验结果表明,本研究成功构建了整合型诱导表达GH载体并能实现GH的可控表达,本研究为以后制备可控表达GH转基因动物、进一步研究GH对机体影响奠定基础。

关键词：诱导表达猪生长激素整合型真核表达载体

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克隆波尔山羊的微卫星DNA鉴定

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中国药科大学学报 2005年第1期36卷 69-72页

作者：刘建唐慧林杨跃飞鞠辉明成勇国家质量监督检验检疫总局动植物检疫实验所北京100029 中国药科大学生命科学与技术学院南京210009 扬州大学兽医学院扬州225009

目的 :利用微卫星DNA技术进行体细胞克隆后获得个体的鉴定。方法 :提取正常山羊、供核波尔山羊、本地受体山羊及克隆个体的基因组DNA ,通过设计微卫星DNA多态PCR引物扩增微卫星DNA序列并对之进行微卫星DNA鉴定。结论 :通过微卫星DNA序... 详细信息

目的 :利用微卫星DNA技术进行体细胞克隆后获得个体的鉴定。方法 :提取正常山羊、供核波尔山羊、本地受体山羊及克隆个体的基因组DNA ,通过设计微卫星DNA多态PCR引物扩增微卫星DNA序列并对之进行微卫星DNA鉴定。结论 :通过微卫星DNA序列的扩增 ,证明克隆个体为供核波尔山羊的克隆。

关键词：体细胞克隆微卫星DNA分析鉴定

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Catalytic Performance of Carbon Smoke over Ag-LSCF Composite Catalysts

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Journal of Wuhan University of Technology(Materials Science) 2025年第1期40卷 30-34页

作者： GUO Guanlun HAN ming LU Shaomin YU Jing ju Hongling 无 Hubei Key Laboratory of Advanced Technology for Automotive Components&Hubei Collaborative Innovation Center for Automotive Components Technology Wuhan University of TechnologyWuhan 430070China College of Mechanical and Transportation Southwest Forestry UniversityKunming 650000China

To improve the catalytic performance of La_(0.6)Sr_(0.4)Co_(0.2)Fe_(0.8)O_(3)(LSCF)towards carbon soot,we utilized the impregnation method to incorporate Ag into the prepared LSCF *** conducted a series of characterization tests and evaluated the soot catalytic activity of the composite catalyst by comparing it with the LaCoO_(3) group,LaFeO_(3) group,and catalyst-free *** results indicate that the Ag-LSCF composite catalyst exhibits the highest soot catalytic activity,with the characteristic temperature values of 376.3,431.1,and 473.9℃at 10%,50%,and 90%carbon soot conversion,*** values are 24.8,20.2,and 23.1℃lower than those of the LSCF *** also shows that LSCF can improve the catalytic activity of soot after compounding with Ag,and reflects the necessity of using catalysts in soot combustion *** characterization and BET test show that Ag-LSCF has more abundant surface-adsorbed oxygen species,larger specific surface area and pore volume than LSCF,which also proves that Ag-LSCF has higher soot catalytic activity.

关键词： metallic composites carbon smoke oxidation perovskite catalyst soot

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Automatic and Rapid Throughflow-Based Optimal Design Method for Multi-Stage Axial-Flow Compressors

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Journal of Thermal Science 2025年第1期34卷 77-91页

作者： LEI Fan ZHANG Xiawen ju Yaping ZHANG Chuhua Department of Fluid Machinery and Engineering School of Energy and Power EngineeringXi'an Jiaotong UniversityXi'an 710049China State Key Laboratory for Strength and Vibration of Mechanical Structures Xi'an Jiaotong UniversityXi'an 710049China Production and Environmental Department China Huaneng Group.Co.Ltd.Beijing 100031China

Throughflow design has the advantages of less time consumption and large optimization space,and thus is the corner stone of advanced design system of multi-stage axial-flow *** majority of relevant studies were limited to the throughflow inverse designs,and quite few works have been till now devoted to the throughflow optimal *** this work,an automatic and rapid throughflow-based optimal design method is proposed for axial-flow compressors in which a throughflow inverse design solver is embedded in optimal genetic algorithm to improve the design efficiency of axial-flow *** types of design parameters in the throughflow inverse design of axial-flow compressors,i.e.,swirl and shroud curve,are simultaneously used to optimize both the blade shape and flow *** proposed method is validated by the redesign optimization of the benchmark axial-flow compressor NASA Stage 35,and the CFD predictions show that the throughflow-based optimization leads to 1.18% efficiency benefit at design *** proposed method is then utilized to the two-dimensional throughflow optimal design of a large-scale 6.5-stage axial-flow industrial *** optimal design results are confirmed by CFD predictions,indicating that the proposed method can effectively improve the design adiabatic efficiency of the compressors by 1.09% within a few minutes on desk-top *** throughflow design implications are also obtained for advanced axial-flow industrial *** work could enhance the capability of throughflow design method and has engineering application value to explore the throughflow optimization space of multi-stage axial-flow compressors.

关键词： multi-stage axial-flow compressor throughflow design empirical correlations genetic algorithm aerodynamic optimization

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Identifying adipocyte-derived exosomal miRNAs as potential novel prognostic markers for radiotherapy of esophageal squamous cell carcinoma

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World Journal of Gastrointestinal Oncology 2025年第2期17卷 211-229页

作者： Yang-Yang Ge Xiao-Chun Xia An-Qing Wu Chen-Ying Ma Ling-hui Yu ju-Ying Zhou Department of Radiation Oncology The First Affiliated Hospital of Soochow UniversitySuzhou 215006Jiangsu ProvinceChina Department of Radiotherapy The Affiliated Tumor Hospital of Nantong UniversityNantong 226361Jiangsu ProvinceChina School of Radiation Medicine and Protection Soochow UniversitySuzhou 215123Jiangsu ProvinceChina Department of Brachytherapy The Affiliated Tumor Hospital of Nantong UniversityNantong 226361Jiangsu ProvinceChina

BACKGROUND Radiation resistance limits radiotherapy efficacy in esophageal squamous cell carcinoma(ESCC).The tumor microenvironment,particularly adipocytes,plays a role in promoting cancer *** vesicles and microRNAs(miRNAs)regulate gene expression and hold prognostic potential for esophageal *** radioresistance mechanisms and identifying radiosensitization targets can help enhance radiotherapy efficacy for esophageal *** To investigate the potential role of miRNAs derived from adipocyte exosomes as prognostic markers for radiotherapy efficacy in *** Free adipocytes were isolated from human thoracic adipose tissue.A co-culture model of adipocytes and ESCC cells was established to observe colony formation and cell survival *** cell apoptosis was assessed by flow *** Blot and immunofluorescence assays were performed to evaluate DNA damage in ESCC cells ***-derived exosomes were isolated by ultracentrifugation and identified by electron microscopy.A similar set of experiments was performed on ESCC cells to analyze cell survival,apoptosis,and DNA damage post-radiation *** from adipose tissue and serum exosomes from ESCC patients pre-and post-radiotherapy were subjected to high-throughput miRNA-sequencing and validated using real-time quantitative polymerase chain *** correlation between potential target miRNAs and the short-term prognosis of radiotherapy in ESCC was evaluated by receiver operating characteristic curve *** Co-culturing adipocytes with ESCC cells enhanced radioresistance,as evidenced by increased colony *** co-culture reduced ESCC cell apoptosis and DNA damage ***-derived exosomes similarly conferred radioresistance in ESCC cells,decreasing apoptosis and DNA damage *** miRNA-sequencing identified miR-660-5p in serum and adipose tissue ***

关键词： Esophageal squamous cell carcinoma Adipocyte Exosomes MicroRNA Radiotherapy

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猪microRNA-378-1过表达载体构建及细胞水平验证

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农业生物技术学报 2012年第8期20卷 942-947页

作者：李莹鞠辉明白立景李奎敖红中国农业科学院北京畜牧兽医研究所

microRNA对细胞的增殖和分化有着重要的调节作用,从大白猪(Susscrofa)基因组DNA上扩增microRNA-378-1前体序列,经XhoⅠ酶切后回收相应片段,连接到pEGP-miR载体中,构建重组载体pEGP-miR-378-1,转染猪胎儿成纤维细胞系(PEF),通过荧光观察... 详细信息

microRNA对细胞的增殖和分化有着重要的调节作用,从大白猪(Susscrofa)基因组DNA上扩增microRNA-378-1前体序列,经XhoⅠ酶切后回收相应片段,连接到pEGP-miR载体中,构建重组载体pEGP-miR-378-1,转染猪胎儿成纤维细胞系(PEF),通过荧光观察转染效率及报告基因的表达效率,利用实时荧光定量PCR(QRT-PCR)测定细胞内microRNA-378-1的表达活性。结果表明,成功地构建了重组载体pEGP-miR-378-1;荧光观察转染后的细胞,显示有较高的转染效率,报告基因有较高的表达效率;QRT-PCR结果显示,实验组和对照组microRNA-378-1表达显著提高,实验组和对照组细胞间差异极显著(P<0.01);为制备转基因动物研究microRNA-378-1功能提供技术平台。

关键词： microRNA-378-1 microRNA 真核表达载体

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