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Establishment of cell lines with porcine spermatogonial stem cell properties

Journal of Animal Science and Biotechnology 2020年第3期11卷 678-689页

作者： Yi Zheng tongying feng Pengfei Zhang Peipei Lei Fuyuan Li Wenxian Zeng Key Laboratory for Animal Genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100ShaanxiChina

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.

关键词： Immortalization Pig Self-renewal Spermatogonial stem cells SV40 large T antigen

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Rational Construction of Hierarchically Porous Fe–Co/N-Doped Carbon/rGO Composites for Broadband Microwave Absorption

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Nano-Micro Letters 2019年第4期11卷 337-352页

作者： Shanshan Wang Yingchun Xu Ruru Fu Huanhuan Zhu Qingze Jiao tongying feng Caihong feng Daxin Shi Hansheng Li Yun Zhao School of Chemistry and Chemical Engineering Beijing Institute of TechnologyZhuhai 100081People’s Republic of China School of Materials and the Environment Beijing Institute of TechnologyZhuhai 519085People’s Republic of China

Developing lightweight and broadband microwave absorbers for dealing with serious electromagnetic radiation pollution is a great challenge.Here,a novel Fe-Co/N-doped carbon/reduced graphene oxide(Fe-Co/NC/rGO)composite with hierarchically porous structure was designed and synthetized by in situ growth of Fe-doped Cobased metal organic frameworks(Co-MOF)on the sheets of porous cocoon-like rGO followed by calcination.The Fe-Co/NC composites are homogeneously distributed on the sheets of porous rGO.The Fe-Co/NC/rGO composite with multiple components(Fe/Co/NC/rGO)causes magnetic loss,dielectric loss,resistance loss,interfacial polarization,and good impedance matching.The hierarchically porous structure of the Fe-Co/NC/rGO enhances the multiple reflections and scattering of microwaves.Compared with the Co/NC and Fe-Co/NC,the hierarchically porous Fe-Co/NC/rGO composite exhibits much better microwave absorption performances due to the rational composition and porous structural design.Its minimum reflection loss(RLmin)reaches?43.26 dB at 11.28 GHz with a thickness of 2.5 mm,and the effective absorption frequency(RL≤?10 dB)is up to 9.12 GHz(8.88-18 GHz)with the same thickness of 2.5 mm.Moreover,the widest effective bandwidth of 9.29 GHz occurs at a thickness of 2.63 mm.This work provides a lightweight and broadband microwave absorbing material while offering a new idea to design excellent microwave absorbers with multicomponent and hierarchically porous structures.

关键词： Fe-doped Co-MOF Hierarchically porous rGO Broadband Microwave absorption performance

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Meclofenamic acid represses spermatogonial proliferation through modulating m^6A RNA modification

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Journal of Animal Science and Biotechnology 2019年第4期10卷 919-930页

作者： Tao Huang Jiayin Guo Yinghua Lv Yi Zheng tongying feng Qiang Gao Wenxian Zeng Key Laboratory of Animal Genetics Breeding and Reproduction of Shaanxi Province College of Animal Science and Technology Northwest A&F University

Background: N6-Methyladenosine(m^6A), the most prevalent modification in mammalian m RNA, plays important roles in numerous biological processes. Several m^6A associated proteins such as methyltransferase like 3(METTL3),methyltransferase like 14(METTL14), α-ketoglutarate-dependent dioxygenase Alk B homolog 5(ALKBH5) and YTH domain containing 2(YTHDC2) are involved in the regulation of spermatogenesis and oogenesis. However, the role of the first detected m^6A demethylase, fat mass and obesity associate protein(FTO), in germ cells remains elusive.Elucidation of FTO roles in the regulation of germ cell fate will provide novel insights into the mammalian reproduction.Methods: Mouse GC-1 spg cells were treated with the ester form of meclofenamic acid(MA2) to inhibit the demethylase activity of FTO. The cellular m^6A and m^6Amlevel were analyzed through high performance liquid chromatography combined with tandem mass spectrometry(HPLC/MS-MS). The cell apoptosis was detected via TUNEL and flow cytometry. The cell proliferation was detected through Ed U and western blot. The m RNA level of core cyclin dependent kinases(CDKs) was quantified via q-PCR. RNA decay assay were performed to detect RNA stability. Dual fluorescence assay was conducted to study whether MA2 affects the expression of CDK2 dependent on the m^6A modification at 3’UTR.Results: MA2 significantly increased the cellular m^6A level and down-regulated the expression of CDK1, CDK2, CDK6 and Cd C25 a, resulting in arrest of G1/S transition and decrease of cell proliferation. MA2 downregulated CDK2 m RNA stability. Additionally, mutation of the predicted m^6A sites in the Cdk2–3’UTR could mitigated the degradation of CDK2 m RNA after MA2 treatment.Conclusion: MA2 affected CDKs expression through the m^6A-dependent m RNA degradation pathway, and thus repressed spermatogonial proliferation.

关键词： Cell cycle FTO Meclofenamic acid N6-methyladenosine Spermatogonial proliferation

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Profiling of miRNAs in porcine Sertoli cells

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Journal of Animal Science and Biotechnology 2021年第1期12卷 236-249页

作者： Xiaoxu Chen Yi Zheng Xueliang Li Qiang Gao tongying feng Pengfei Zhang Mingzhi Liao Xiu’e Tian Hongzhao Lu Wenxian Zeng College of Biological Science and Engineering Shaanxi University of TechnologyHanzhong 723001China Key Laboratory of Animal Genetics Breeding and Reproduction of the Ministry of AgricultureCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100ShaanxiChina College of Life Science Northwest A&F UniversityYangling 712100ShaanxiChina.

Background: Sertoli cells(SCs) create a specialized environment to support and dictate spermatogenesis.MicroRNAs(miRNAs), a kind of ~ 22 nt small noncoding RNAs, have been reported to be highly abundant in mouse SCs and play critical roles in spermatogenesis. However, the miRNAs of porcine SCs remain largely unknown.Methods: We isolated porcine SCs and conducted small RNA sequencing. By comparing miRNAs in germ cells, we systematically analyzed the miRNA expression pattern of porcine SCs. We screened the highly enriched SC miRNAs and predicted their functions by Gene Ontology analysis. The dual luciferase assay was used to elucidate the regulation of tumor necrosis factor receptor(TNFR)-associated factor 3(TRAF3) by ssc-miR-149.Results: The analysis showed that 18 miRNAs were highly expressed in SCs and 15 miRNAs were highly expressed in germ cells. These miRNAs were predicted to mediate SC and germ cell functions. In addition, ssc-miR-149 played critical roles in SCs by targeting TRAF3.Conclusion: Our findings provide novel insights into the miRNA expression pattern and their regulatory roles of porcine SCs.

关键词： MicroRNAs Pig Sertoli cell Spermatogenesis TRAF3

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基于应用型大学的“以学生为中心”的无机化学教学设计与实践

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云南化工 2023年第6期50卷 200-204页

作者：冯彤英吴灵魏永春马文英北京理工大学珠海学院材料与环境学院广东珠海519088

“以学生为中心”就是注重学生的学习动力及思维方式,关注学生的学习过程、提高学生的学习兴趣和学习能力。基于这一理念,针对应用型本科学校—北京理工大学珠海学院无机化学课程教学中普遍存在的“痛点”问题,从教学模式的调整、教学... 详细信息

“以学生为中心”就是注重学生的学习动力及思维方式,关注学生的学习过程、提高学生的学习兴趣和学习能力。基于这一理念,针对应用型本科学校—北京理工大学珠海学院无机化学课程教学中普遍存在的“痛点”问题,从教学模式的调整、教学内容的优化与教学资源的建设、教学活动的创新设计,以及考核方式的完善等几个方面进行了教学改革,以期解决目前无机化学教学中出现的问题,达到培养学生能力的目的。

关键词：以学生为中心无机化学教学问题教学设计过程考核

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大熊猫脐带间充质干细胞外泌体的分离鉴定及miRNAs富集分析

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兽类学报 2023年第1期43卷 33-40页

作者：安俊辉李媛王东辉陈佳松李红艳梁小虎封同英蔡志刚侯蓉曾长军刘玉良四川农业大学动物科技学院畜禽遗传资源发掘与创新利用四川省重点实验室成都611130 成都大熊猫繁育研究基地四川省濒危野生动物保护生物学重点实验室成都610081 四川省大熊猫科学研究院成都610081

大熊猫脐带间充质干细胞(Umbilical cord mesenchymal stem cells,UC-MSCs)通过旁分泌所释放的外泌体在大熊猫保健与疾病治疗方面具有一定的应用前景。本研究旨在建立大熊猫UC-MSCs外泌体分离方法,开展生物学特征分析和分子鉴定,并研究U... 详细信息

大熊猫脐带间充质干细胞(Umbilical cord mesenchymal stem cells,UC-MSCs)通过旁分泌所释放的外泌体在大熊猫保健与疾病治疗方面具有一定的应用前景。本研究旨在建立大熊猫UC-MSCs外泌体分离方法,开展生物学特征分析和分子鉴定,并研究UC-MSCs外泌体中miRNAs的种类与功能。采用超速离心法从大熊猫UCMSCs培养上清中成功分离外泌体,通过透射电子显微镜进行形态学观察,纳米颗粒跟踪分析仪检测粒径大小,蛋白免疫印迹法检测特异性分子标记表达。采用Small RNA测序技术对UC-MSCs外泌体中的miRNAs进行鉴定,并对其靶基因进行了预测与功能分析。结果显示,大熊猫UC-MSCs外泌体呈圆形杯托状结构,直径为(79.15±4.81)nm,外泌体标志蛋白CD81与TSG101呈阳性表达而CALNEXIN呈阴性表达。大熊猫UC-MSCs外泌体中的miRNA主要为miR-148-3p(30.28%)与miR-21-5p(21.72%)。本研究首次从大熊猫UC-MSCs培养上清中分离出外泌体,并对其所含的miRNAs进行富集分析及功能预测,为大熊猫UC-MSCs外泌体后续的功能研究及应用奠定基础。

关键词：大熊猫脐带间充质干细胞外泌体 miRNA

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独立学院物理化学实验教学实践与探索

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广东化工 2013年第18期40卷 164-165页

作者：冯彤英吕京美詹世景北京理工大学珠海学院广东珠海519088

论文结合独立学院的特点、培养目标,从实验内容、教学方式、考核内容等方面探讨了物理化学实验的教学实践以及教学过程中出现的问题。

关键词：独立学院物理化学实验教学实践

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浅析独立学院大学生自主学习能力的培养——以无机化学课程教学为例

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广东化工 2016年第12期43卷 286-287页

作者：冯彤英马文英魏永春北京理工大学珠海学院广东珠海519088

文章针对独立学院的特点,结合无机化学教学实践从学习兴趣、课程结构、教学手段、教育技术等方面探讨大学生自主学习能力的培养。

关键词：独立学院无机化学自主学习能力

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基于应用型人才培养的无机化学实验教学改革探索

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广东化工 2019年第1期46卷 196-197页

作者：冯彤英魏永春詹世景吕京美马文英蔡阳伦北京理工大学珠海学院广东珠海519085

应用型本科人才培养已成为高等教育发展的必然趋势,本文基于应用型人才培养要求,从教学内容、教学方法以及考核机制三个方面对现行的无机化学实验教学进行了改革探索。教学实践表明,这些改革有利于促进学生学习无机化学实验的兴趣和积... 详细信息

应用型本科人才培养已成为高等教育发展的必然趋势,本文基于应用型人才培养要求,从教学内容、教学方法以及考核机制三个方面对现行的无机化学实验教学进行了改革探索。教学实践表明,这些改革有利于促进学生学习无机化学实验的兴趣和积极性、提高实验教师的教学质量以及增强学生的专业技能及动手能力。

关键词：无机化学实验应用型人才教学探索

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Judgement on Extended Hepatectomy for Primary Liver Cancer

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The Chinese-German Journal of Clinical Oncology 2003年第3期2卷 136-139,189页

作者： YANGJiamei tongying CHENHan XUfeng KANTong WUMengchao EasternHepatobiliarySurgeryHospital SecondMilitaryMedicalUniversityShanghai200438China

Objective: To expore the indications and safety of extended hepatectomy for primary liver cancer (PLC). Methods: From Nov. 2000 to Oct. 2002, 33 patients with PLC received extended hepatectomy, 26 of whom were complicated with liver cirrhosis. Preoperative findings, intraoperative management and the outcome of postoperative recovery were analyzed and evaluated. Results: Operative mortality was zero in this group and postoperative morbidity of complications was 33.3%, and all complications were cured after proper treatment. Conclusion: Extended hepatectomy for PLC was safe and viable for the patients who had enough reserve of liver function if the incised liver volume was less than 50% of the liver and the supporting treatment of the remnant liver was given as well as the postoperative complications were managed appropriately.

关键词：肝切除术原发性肝癌切除范围手术治疗

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