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Molecular cloning and identification of mouse epididymis-specific gene mHongl, the homologue of rat HongrES1

Asian Journal of Andrology 2012年第4期14卷 626-634页

作者： shuang-gang hu Han Du Guang-Xin Yao Yong-Lian Zhang Shanghai Key Laboratory for Molecular Andrology State Key Laboratory of Molecular BiologyInstitute of Biochemistry and Cell BiologyShanghai Institutes for Biologica SciencesChinese Academy of SciencesShanghai 200031China Shanghai Institute of Planned Parenthood Research Shanghai 200031China

Previous studies have shown that rat epididymis-specific gene HongrES1 plays important roles in sperm capacitation and fertility. In this study, we cloned the mouse homologue gene by sequence alignment and RT-PCR methods and designated it as mHongl. The mHongl gene is located on chromosome 12p14, spanning five exons. The cDNA sequence consists of 1257 nucleotides and encodes a 419 amino-acid protein with a predicted N-terminal signal peptide of 20 amino acids. The mHongl mRNA shows similarity with HongrES1 in the expression patterns：（i） specific expression in epididymal tissue, especially in the cauda region; and （ii） androgen-dependence but testicular fluid factor independence. Its protein product shows 71% similarity with HongrES 1 and contains a classical serpin domain as does HongrES1. A polyclonal antibody against mHongl with high specificity and sensitivity was raised. Like HongrES1, the mHongl protein shows a checker-board expression pattern in the epididymal epithelium and is secreted into the epididymal lumen. The mHongl protein shows higher glycosylation than HongrES1. Although both of them are deposited onto the sperm head surface, mHongl is localized to the equatorial segment, which is different from that of HongrES 1. The mHongl protein can be removed from the sperm membrane by high ionic strength and therefore can be classed as an extrinsic membrane protein. Collectively, we conclude that mHongl is the homologue of HongrES1 and the present work paves the way for establishing animal models to elucidate the precise functions of HongrES1 and mHongl.

关键词： androgen epididymal secretory protein epididymis glycosylation sperm maturation

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Isolation and characterization of detergent-resistant membranes from rat spermatozoa

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Asian Journal of Andrology 2014年第5期16卷 790-791页

作者： shuang-gang hu Xiang-Qi Li Chun-hua Tang Yun Sun Yong-Lian Zhang Department of Reproductive Medicine Renji Hospital School of Medicine Shanghai Jiaotong University Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics Shanghai China Shanghai Key Laboratory for Molecular Andrology State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Dear Editor We present herein the isolation and characterization of detergent-resistant membranes （DRMs） from rat spermatozoa. This is the first study on rat sperm DRMs. Detergent-resistant membranes including lipid raft and caveolae are cholesterol- and sphingolipid-rich microdomains of the plasma membrane that concentrate components of certain signal transduction pathways. They are receiving increasing attention as devices involved in the regulation of membrane fluidity and membrane protein trafficking. Until now, DRMs have been identified in spermatozoa from different species, including human, mouse, porcine and bovine. Increasing evidence has shown that these microdomains play important roles during sperm maturation in the epididymis and serve as platforms for the assembly of key recognition molecules that are involved in sperm-zona pellucida interaction on the sperm surface during capacitation. However, until now, no any studies on rat sperm DRMs have been reported. In the present work, we focused on the biochemical isolation and characterization of rat sperm DRMs.

关键词： Isolation characterization detergent-resistant

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The expression of the new epididymal luminal protein of PDZ domain containing 1 is decreased in asthenozoospermia

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Asian Journal of Andrology 2018年第2期20卷 154-159页

作者： A-Juan Liang Gui-Shuan Wang Ping Ping shuang-gang hu Yu Lin Yi Ma Zheng-Zheng Duan Han-Shu Wang Fei Sun International Peace Maternity and Child Health Hospital Shanghai Key Laboratory for Reproductive Medicine Institute of Embryo-Fetal Original Adult Disease School of Medicine Shanghai Jiao Tong University Shanghai 200030 China Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics Center for Reproductive Medicine Renji Hospital School of Medicine Shanghai Jiao Tong University Shanghai 200135 China Medical School Institute of Reproductive Medicine Nantong University Nantong 226001 China

Spermatozoa are not mature until they transit the epididymis where they acquire motility and the ability to fertilize an egg through sequential modifications. The epididymis has three functional regions, caput, corpus, and cauda, and the luminal proteins of the epididymis play important roles in the above modifications. However, the proteins with differential enrichment between the caput and cauda are still largely unknown. To reveal the functions of the caput and cauda during sperm maturation, luminal proteins from caput and cauda of mice were analyzed by isobaric tag for relative and absolute quantitation （iTRAQ）. Overall, 128 differentially enriched proteins were found, of which 46 were caput enriched and 82 were cauda enriched. Bioinformatic analysis showed that lipid metabolism was active in the caput; while anion- and cation-binding activity and phosphorus and organophosphate metabolism were active in the cauda. A new epididymal luminal protein, the caput-enriched PDZ domain containing 1 （Pdzkl）, also named Na^＋/H^＋ exchange regulatory cofactor 3 （NHERF3）, which plays a critical role in cholesterol metabolism and carnitine transport, was found in the lipid metabolism. Western blotting and immunofluorescence analyses showed that Pdzkl was expressed in the epididymis but not in the testis, and localized at the middle piece of the sperm tail. Pdzkl protein level was also reduced in the spermatozoa in case of asthenozoospermic patients compared with that in normozoospermic men, suggesting that Pdzkl may participate in sperm maturation regulation and may be associated with male infertility. These results may provide new insights into the mechanisms of sperm maturation and male infertility.

关键词： asthenozoospermia epididymis fertility PDZ domain containing 1 sperm

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精子表面的GPI锚定蛋白

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生殖与避孕 2008年第1期28卷 36-40页

作者：邹美胡双纲胡建民沈阳农业大学沈阳110161 中国科学院上海生命科学研究院生物化学与细胞生物学研究所上海200031

糖基化磷脂酰肌醇锚定蛋白(GPI-APs)是一类通过其羧基末端的糖基化磷脂酰肌醇结构锚定于真核细胞膜表面而不跨越其磷脂膜双层的蛋白。精子上的GPI-APs是一类重要的精子表面蛋白,已报道的有十几种,功能多样,在精子的发生、成熟、获能、... 详细信息

糖基化磷脂酰肌醇锚定蛋白(GPI-APs)是一类通过其羧基末端的糖基化磷脂酰肌醇结构锚定于真核细胞膜表面而不跨越其磷脂膜双层的蛋白。精子上的GPI-APs是一类重要的精子表面蛋白,已报道的有十几种,功能多样,在精子的发生、成熟、获能、受精以及免疫保护等方面起着重要作用。这类蛋白有些可能是某些男性不育症潜在的诊断和治疗靶点,有些则是有前景的免疫避孕候选靶标。

关键词：糖基化磷脂酰肌醇锚定蛋白 GPI-APs 精子免疫避孕

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Monoclonal Antibodies Reveal Novel Localization of SPAG11E in Spermatozoa and the Antifertility Potential of SPAG11E Motifs

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Reproductive and Developmental Medicine 2018年第4期2卷 191-200,I0001-I0005页

作者： Zhi-Kai Wang Yi-Ting Yang Xin-Yu Chen shuang-gang hu Ping Zhu Wan-Xiang Xu Li Ma He-Guo Yu hua Diao Yong-Lian Zhang Clinical Laboratory(Hospital of the SIPPR)and NHC Key Laboratory of Reproduction Regulation Shanghai Institute of Planned Parenthood ResearchShanghai 200032China Core Facility of Basic Medical Sciences School of MedicineShanghai Jiao Tong UniversityShanghai 200025China Shanghai Key Laboratory of Molecular Andrology State Key Laboratory of Molecular BiologyInstitute of Biochemistry and Cell BiologyChinese Academy of SciencesShanghai 200031China

Objective: SPAG11E is the first β-defensin that has been reported to activate Ca;uptake and sperm motility. However, the exact subcellular localization and interaction of SPAG11E with sperm remain controversial because of the lack of qualified antibody tools. SPAG11E is also a potential male antifertility target because SPAG11E fragment conjugated with a carrier protein exhibits male contraceptive vaccine ***, the fine B-cell epitope motifs of SPAG11E have not been analyzed, which hampered further exploration of the potential ***: Polyclonal and monoclonal antibodies（mcAbs） of mature SPAG11E were raised and qualified with Western blotting. Subcellular localization of SPAG11E was revealed by Western blotting, immunohistochemistry staining, and electron microscopy. B-cell epitopes of rat SPAG11E were mapped by Western blotting using polyclonal and mcAbs. Based on the conservation of the identified epitope motifs between rat and mouse SPAG11E, antifertility potential of the epitope motifs was evaluated by the offspring of the males compromised with specific ***: SPAG11E antibodies of high quality were obtained and all B-cell epitope motifs of rat SPAG 11 E were mapped, in which conserved epitope motifs of SPAG11E in various species were discovered. The epitope motifs recognized by mcAbs were identified respectively. With mcAbs, rat SPAG11E was proved to be expressed in the caput region of epididymis. A novel finding was that SPAG 11 E was located in the flagella and nuclei of sperm as revealed by immunoelectron microscopy. In addition, the males treated with mcAbs（3#-1 and 10#B4） showed apparently fewer ***: SPAG11E revealed a β-defensin with novel localization in sperm flagellum and nucleus with qualified antibodies. All B-cell epitope motifs of rat SPAG11E were determined, and the antifertility potential was proved by corresponding mcAbs.

关键词： Antibody Contraception Epitope SPAG11E Sperm

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Effect of Nb-V microalloying on the hot deformation behavior of medium Mn steels

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International Journal of Minerals,Metallurgy and Materials 2025年第2期32卷 360-368页

作者： Yumeng Wang Qinyi Guo Bin hu Haiwen Luo School of Metallurgical and Ecological Engineering University of Science and Technology BeijingBeijing 100083China

The influence of Nb-V microalloying on the hot deformation behavior and microstructures of medium Mn steel(MMS)was investigated by uniaxial hot compression *** establishing the constitutive equations for simulating the measured flow curves,we successfully constructed deformation activation energy(Q)maps and processing maps for identifying the region of flow *** concluded the following consequences of Nb-V alloying for MMS.(i)The critical strain increases and the increment diminishes with the increasing deformation temperature,suggesting that NbC precipitates more efficiently retard dynamic recrystallization(DRX)in MMS compared with solute Nb.(ii)The deformation activation energy of MMS is significantly increased and even higher than that of some reported high Mn steels,suggesting that its ability to retard DRX is greater than that of the high Mn content.(iii)The hot workability of MMS is improved by narrowing the hot processing window for the unstable flow stress,in which fine recrystallized and coarse unrecrystallized grains are present.

关键词： medium manganese steel niobium-vanadium microalloying flow behavior dynamic recrystallization hot workability

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Co/Co_(7)Fe_(3)heterostructures with controllable alloying degree on carbon spheres as bifunctional electrocatalyst forrechargeable zinc-air batteries

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International Journal of Minerals,Metallurgy and Materials 2025年第2期32卷 476-487页

作者： Junkang Chen Yongyue Zhuang Yanxin Qiao Yu Zhang Aihua Yuan hu Zhou School of Materials Science and Engineering Jiangsu University of Science and TechnologyZhenjiang 212003China School of Environmental and Chemical Engineering Jiangsu University of Science and TechnologyZhenjiang 212003China

Exploring efficient and nonprecious metal electrocatalysts of oxygen reduction reaction(ORR)and oxygen evolution reaction(OER)is crucial for developing rechargeable zinc-air batteries(ZABs).Herein,an alloying-degree control strategy was employed to fabricate nitrogen-doped carbon sphere(NCS)decorated with dual-phase Co/Co_(7)Fe_(3)heterojunctions(CoFe@NCS).The phase composition of materials has been adjusted by controlling the alloying *** optimal CoFe_(0.08)@NCS electrocatalyst displays a half-wave potential of 0.80 V for ORR and an overpotential of 283 mV at 10 mA·cm^(-2)for OER in an alkaline *** intriguing bifunctional electrocatalytic activity and durability is attributed to the hierarchically porous structure and interfacial electron coupling of highly-active Co_(7)Fe_(3)alloy and metallic Co *** the CoFe_(0.08)@NCS material is used as air-cathode catalyst of rechargeable liquid-state zinc-air battery(ZAB),the device shows a high peak power-density(157 mW·cm^(-2))and maintains a stable voltage gap over 150 h,outperforming those of the benchmark(Pt/C+RuO_(2))-based *** particular,the as-fabricated solid-state flexible ZAB delivers a reliable compatibility under different bending *** work provides a promising strategy to develop metal/alloy-based electrocatalysts for the application in renewable energy conversion technologies.

关键词： bifunctional electrocatalysts oxygen reduction reaction oxygen evolution reaction zinc-air battery metal/alloy carbon sphere

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Photo‑Energized MoS_(2)/CNT Cathode for High‑Performance Li–CO_(2)Batteries in a Wide‑Temperature Range

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Nano-Micro Letters 2025年第1期17卷 160-175页

作者： Tingsong hu Wenyi Lian Kang hu Qiuju Li Xueliang Cui Tengyu Yao Laifa Shen Jiangsu Key Laboratory of Materials and Technologies for Energy Storage College of Materials Science and TechnologyNanjing University of Aeronautics and AstronauticsNanjing 210016People’s Republic of China

Li–CO_(2) batteries are considered promising energy storage systems in extreme environments such as Mars;however,severe performance degradation will occur at a subzero temperature owning to the sluggish reaction ***,a photo-energized strategy adopting sustainable solar energy in wide working temperature range Li–CO_(2) battery was achieved with a binder-free MoS_(2)/carbon nanotube(CNT)photo-electrode as *** unique layered structure and excellent photoelectric properties of MoS_(2) facilitate the abundant generation and rapid transfer of photo-excited carriers,which accelerate the CO_(2) reduction and Li_(2)CO_(3) decomposition upon *** illuminated battery at room temperature exhibited high discharge voltage of 2.95 V and mitigated charge voltage of 3.27 V,attaining superior energy efficiency of 90.2%and excellent cycling stability of over 120 *** at an extremely low temperature of−30℃,the battery with same electrolyte can still deliver a small polarization of 0.45 V by the photoelectric and photothermal synergistic mechanism of MoS_(2)/CNT *** work demonstrates the promising potential of the photo-energized wide working temperature range Li–CO_(2) battery in addressing the obstacle of charge overpotential and energy efficiency.

关键词： Li-CO_(2)batteries Photo-energized Wide operation-temperature Kinetics MoS_(2)

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CALCULATION OF NAMBU MECHANICS

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Journal of Computational Mathematics 2006年第3期24卷 444-450页

作者： shuang-hu Wang The State Key Laboratory of Computational Physics Institute of Applied Physics and Computational Mathematics Beijing 100088 China

In this paper, a fundamental fact that Nambu mechanics is source free was proved. Based on this property, and via the idea of prolongation, finite dimensional Nambu system was prolonged to difference jet bundle. Structure preserving numerical methods of Nambu equations were established. Numerical experiments were presented at last to demonstrate advantages of the structure preserving schemes.

关键词： Nambu equations Differential forms Difference schemes Structure preserving methods.

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反复IVF失败患者卵泡液代谢组学的检测分析

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生殖与避孕 2012年第4期32卷 248-253页

作者：夏兰赵晓明孙赟洪燕郑菊芬向祖琼姚宁赵磊文郭毅胡双纲上海交通大学医学院附属仁济医院生殖医学科上海200001

目的:探讨与卵子发育潜能相关的代谢物质。方法:运用气相色谱-质谱仪(GC-MS)回顾性分析13例反复IVF失败(RIF组)和15例同期第1次行IVF-ET成功妊娠患者(对照组)的卵泡液代谢组学。结果:用偏最小二乘法分析样本主成分,结果显示RIF组和对照... 详细信息

目的:探讨与卵子发育潜能相关的代谢物质。方法:运用气相色谱-质谱仪(GC-MS)回顾性分析13例反复IVF失败(RIF组)和15例同期第1次行IVF-ET成功妊娠患者(对照组)的卵泡液代谢组学。结果:用偏最小二乘法分析样本主成分,结果显示RIF组和对照组的主成分积分值集中分布于2个不同区域,且反复IVF失败组较对照组有11种氨基酸含量显著升高,而二元羧酸、胆固醇及部分有机酸含量显著降低。结论:升高的缬氨酸、苏氨酸、异亮氨酸、半胱氨酸、丝氨酸、脯氨酸、丙氨酸、苯丙氨酸、赖氨酸、甲硫氨酸、鸟氨酸,和降低的二元羧酸、胆固醇可能成为预测卵子发育潜能的代谢标志物。

关键词：反复IVF失败卵泡液代谢组学胚胎发育潜能

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