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学者姓名：朱婷
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Abstract ：
Duck adenovirus 3 (DAdV-3) causes liver damage and bleeding, with morbidity rates ranging from 40 to 55% and mortality rates between 35 and 43%. Co-infection with other pathogens complicates disease control, significantly impacting the duck breeding industry. Currently, there have been no effective vaccines or treatments for DAdV-3. Therefore, rapid, specific, and sensitive detection methods are crucial for preventing and controlling this virus. Our study developed a lateral flow strip (LFS) detection method using recombinase polymerase amplification (RPA) and CRISPR/Cas12a. The RPA-CRISPR/Cas12a-LFS method, performed at 37 degrees C, allowed for result visualization without sophisticated equipment. It targeted the DAdV-3 Fiber-2 gene and achieved a detection limit of 3.0 gene copies. Additionally, this method demonstrated high specificity, with no cross-reactivity to eight other avian viruses. The reaction time of RPA-CRISPR/Cas12a-LFS is only 45 min. Analysis of 95 waterfowl samples showed 98.95% consistency and agreement with quantitative polymerase chain reaction using the Fiber-2 RPA-CRISPR/Cas12a-LFS method. These findings highlighted the potential of this user-friendly, rapid, sensitive, and accurate detection method for on-site DAdV-3 detection.
Keyword ：
CRISPR/Cas12a CRISPR/Cas12a DAdV-3 DAdV-3 LFS LFS on-site detection on-site detection RPA RPA
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| GB/T 7714 | Liang, Qi-Zhang , Chen, Wei , Bi, Yuhai et al. Recombinase polymerase amplification combined with CRISPR/Cas12a technology for rapid on-site detection of duck adenovirus 3 [J]. | FRONTIERS IN MICROBIOLOGY , 2025 , 16 . |
| MLA | Liang, Qi-Zhang et al. "Recombinase polymerase amplification combined with CRISPR/Cas12a technology for rapid on-site detection of duck adenovirus 3" . | FRONTIERS IN MICROBIOLOGY 16 (2025) . |
| APA | Liang, Qi-Zhang , Chen, Wei , Bi, Yuhai , Wang, Weiwei , Liu, Rong-Chang , Fu, Qiu-Ling et al. Recombinase polymerase amplification combined with CRISPR/Cas12a technology for rapid on-site detection of duck adenovirus 3 . | FRONTIERS IN MICROBIOLOGY , 2025 , 16 . |
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Background The spleen is a highly organized lymphoid organ with a critical role in antimicrobial immune responses. Pseudorabies virus (PRV) is widely recognized for its ability to induce immunosuppression, with the spleen being one of the primary parenchymal organs of target. Viral infections often disrupt endoplasmic reticulum (ER) homeostasis, leading to ER stress and subsequent apoptosis. This study aimed to investigate the relationship between PRV-induced spleen damage and ER stress. Results Both classical (Min-A) and variant (SX-2018) PRV strains caused significant histopathological damage in the mouse spleen, including marked reductions in CD8(+) T cell populations and increased lymphocyte apoptosis. Further analyses revealed that PRV infection triggered ER stress and activated the PERK-eIF2 alpha-ATF4-CHOP signaling pathway in the spleen. Notably, treatment with the ER stress inhibitor 4-phenylbutyric acid(4-PBA) mitigated lymphocyte depletion and improved survival rates in PRV-infected mice. Conclusions PRV infection leads to lymphocyte depletion in mouse spleens, closely associated with ER stress and apoptosis.
Keyword ：
4-PBA 4-PBA Apoptosis Apoptosis CD8(+) t cell CD8(+) t cell ER stress ER stress PRV PRV Spleen Spleen
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| GB/T 7714 | Liu, Kun , Xin, Hangkuo , Meng, Kuiyang et al. Pseudorabies virus infection induces lymphocyte depletion associated with cellular apoptosis and endoplasmic reticulum stress in the mouse spleen [J]. | BMC VETERINARY RESEARCH , 2025 , 21 (1) . |
| MLA | Liu, Kun et al. "Pseudorabies virus infection induces lymphocyte depletion associated with cellular apoptosis and endoplasmic reticulum stress in the mouse spleen" . | BMC VETERINARY RESEARCH 21 . 1 (2025) . |
| APA | Liu, Kun , Xin, Hangkuo , Meng, Kuiyang , Zhao, Long , Lin, Shengyu , Chen, Wei et al. Pseudorabies virus infection induces lymphocyte depletion associated with cellular apoptosis and endoplasmic reticulum stress in the mouse spleen . | BMC VETERINARY RESEARCH , 2025 , 21 (1) . |
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CD56作为NK细胞的重要表面分子，在NK细胞的发育、亚群分型和功能发挥等方面发挥重要的作用。因此，为了便于筛选不同亚群的猪NK细胞并探究其生物学功能，本研究制备了猪CD56单克隆抗体并对其抗原表位进行了初步鉴定。首先，通过生物信息软件预测并选取猪CD56蛋白胞外区的优势抗原表位的氨基酸序列(50-499aa),利用原核表达系统表达猪CD56胞外区的优势抗原表位蛋白并进行纯化和复性，之后，将猪CD56重组蛋白免疫BALB/c小鼠，通过细胞融合、亚克隆等试验，获得一株稳定分泌猪CD56单克隆抗体的杂交瘤细胞株，命名为1G8。Western blot结果显示，制备的猪CD56单抗可以与猪脑和脾总蛋白发生特异性结合；为了进一步验证CD56单抗的特异性，作者分离了猪外周血NK细胞，以制备的猪CD56单抗作为一抗进行间接免疫荧光试验，结果显示，制备的单抗能够与NK细胞膜特异性结合。最后，为了进一步探究单抗识别的抗原表位，作者将选取的猪CD56基因进一步截短为四个截短体(50-192 aa、193-294 aa、295-397 aa、398-499 aa)并构建真核表达重组质粒，然后转染至293T细胞进行真核表达，Western blot和间接免疫荧光结果显示，制备的CD56单抗识别的抗原表位为猪CD56蛋白的193-294 aa。综上，本研究成功制备了猪CD56单克隆抗体，并对其特异性及抗原表位进行了初步鉴定，对猪NK细胞亚群的筛选及探究猪NK细胞的生物学功能具有重要意义。
Keyword ：
CD56 CD56 NK细胞 NK细胞 单克隆抗体 单克隆抗体 抗原表位 抗原表位 猪 猪
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| GB/T 7714 | 辛航阔 , 谢青青 , 刘坤 et al. 一株猪CD56单克隆抗体的制备及抗原表位的初步鉴定 [J]. | 畜牧兽医学报 , 2024 , 55 (06) : 2662-2671 . |
| MLA | 辛航阔 et al. "一株猪CD56单克隆抗体的制备及抗原表位的初步鉴定" . | 畜牧兽医学报 55 . 06 (2024) : 2662-2671 . |
| APA | 辛航阔 , 谢青青 , 刘坤 , 林圣宇 , 陈炜 , 郑小惠 et al. 一株猪CD56单克隆抗体的制备及抗原表位的初步鉴定 . | 畜牧兽医学报 , 2024 , 55 (06) , 2662-2671 . |
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【目的】建立新型番鸭细小病毒（New-genotype muscovy duck parvovirus, N-MDPV）荧光RPA恒温检测方法，为基层提供可视化快速检测技术手段。【方法】以N-MDPV的VP3基因保守片段为靶点，使用EXO荧光探针特定结合VP3基因保守片段，设计特异的RPA扩增引物并利用重组酶聚合酶扩增技术扩增目的基因，从而建立一种荧光RPA恒温检测N-MDPV的方法，确定反应体系的最佳反应时间和温度，分析该方法特异性和灵敏性；对收集的病料进行核酸检测，并与传统PCR和病毒分离鉴定方法检测结果进行比较。【结果】建立的荧光RPA恒温检测方法最佳反应温度为39℃，最佳反应时间为30 min；灵敏性高，最低核酸检测限度可达10 fg·μL-1；对新型番鸭细小病毒核酸进行特异性扩增，结果显示对鸭腺病毒3型（Duck adenovirus type 3, DAdV-3）、禽腺病毒4型（Fowl adenovirus serotype 4, FAdV-4）、鸭圆环病毒（Duck circovirus, DuCV）、鸭瘟病毒（Duck plague virus, DPV）、鸭病毒性肝炎病毒（Duck hepatitis virus, DHV）、鸭坦布苏病毒（Duck tembusu virus, DTMUV）和新型鸭呼肠孤病毒（Novel duck reovirus, NDRV）的核酸均未有发生交叉反应，特异性良好。利用本研究建立的RPA快检方法、传统PCR方法以及病毒分离鉴定方法对收集的38份鸭组织病料核酸样品进行检测，结果显示阳性率分别为36.8%(14/38)、36.8%(14/38)和31.6%(12/38);RPA检测后呈阳性的样品经PCR方法检测与病毒分离鉴定方法检测均呈现阳性，阳性符合率为100%。【结论】该方法可以很好地应用于缺乏相应检测设备的基层进行新型番鸭细小病毒的大规模临床样本检测，为新型番鸭细小病毒的可视化快速检测提供技术手段。
Keyword ：
RPA恒温检测 RPA恒温检测 新型番鸭细小病毒 新型番鸭细小病毒 重组酶聚合酶扩增技术 重组酶聚合酶扩增技术
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| GB/T 7714 | 陈炜 , 梁齐章 , 张佳雪 et al. 新型番鸭细小病毒荧光RPA恒温检测方法的建立 [J]. | 福建农业学报 , 2024 , 39 (09) : 1044-1050 . |
| MLA | 陈炜 et al. "新型番鸭细小病毒荧光RPA恒温检测方法的建立" . | 福建农业学报 39 . 09 (2024) : 1044-1050 . |
| APA | 陈炜 , 梁齐章 , 张佳雪 , 焦文龙 , 林永强 , 刘荣昌 et al. 新型番鸭细小病毒荧光RPA恒温检测方法的建立 . | 福建农业学报 , 2024 , 39 (09) , 1044-1050 . |
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BackgroundDuck circovirus (DuCV) infections commonly induce immunosuppression and secondary infections in ducks, resulting in significant economic losses in the duck breeding industry. Currently, effective vaccines and treatments for DuCV have been lacking. Therefore, rapid, specific, and sensitive detection methods are crucial for preventing and controlling DuCV.MethodsA lateral flow strip (LFS) detection method was developed using recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 12a (Cas12a). The RPA-CRISPR/Cas12a-LFS targeted the DuCV replication protein (Rep) and was operated at 37 degrees C and allowed for visual interpretation without requiring sophisticated equipment.ResultsThe results revealed that the reaction time of RPA-CRISPR/Cas12a-LFS is only 45 min. This method achieved a low detection limit of 2.6 gene copies. Importantly, this method demonstrated high specificity and no cross-reactivity with six other avian viruses. In a study involving 97 waterfowl samples, the Rep RPA-CRISPR/Cas12a-LFS showed 100% consistency and agreement with real-time quantitative polymerase chain reaction.ConclusionThese findings underscored the potential of this user-friendly, rapid, sensitive, and accurate detection method for on-site DuCV detection.
Keyword ：
CRISPR/Cas12a CRISPR/Cas12a Duck circovirus (DuCV) Duck circovirus (DuCV) Lateral flow strip (LFS) Lateral flow strip (LFS) On-site detection On-site detection Recombinase polymerase amplification (RPA) Recombinase polymerase amplification (RPA)
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| GB/T 7714 | Liang, Qi-Zhang , Chen, Wei , Liu, Rong-Chang et al. CRISPR/Cas12a and recombinase polymerase amplification-based rapid on-site nucleic acid detection of duck circovirus [J]. | VIROLOGY JOURNAL , 2024 , 21 (1) . |
| MLA | Liang, Qi-Zhang et al. "CRISPR/Cas12a and recombinase polymerase amplification-based rapid on-site nucleic acid detection of duck circovirus" . | VIROLOGY JOURNAL 21 . 1 (2024) . |
| APA | Liang, Qi-Zhang , Chen, Wei , Liu, Rong-Chang , Fu, Qiu-Ling , Fu, Guang-Hua , Cheng, Long-Fei et al. CRISPR/Cas12a and recombinase polymerase amplification-based rapid on-site nucleic acid detection of duck circovirus . | VIROLOGY JOURNAL , 2024 , 21 (1) . |
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Prion diseases are a group of neurodegenerative diseases characterized by mitochondrial dysfunction and neuronal death. Mitophagy is a selective form of macroautophagy that clears injured mitochondria. Prohibitin 2 (PHB2) has been identified as a novel inner membrane mitophagy receptor that mediates mitophagy. However, the role of PHB2 in prion diseases remains unclear. In this study, we isolated primary cortical neurons from rats and used the neurotoxic prion peptide PrP106-126 as a cell model for prion diseases. We examined the role of PHB2 in PrP106-126-induced mitophagy using Western blotting and immunofluorescence microscopy and assessed the function of PHB2 in PrP106-126-induced neuronal death using the cell viability assay and the TUNEL assay. The results showed that PrP106-126 induced mitochondrial morphological abnormalities and mitophagy in primary cortical neurons. PHB2 was found to be indispensable for PrP106-126-induced mitophagy and was involved in the accumulation of PINK1 and recruitment of Parkin to mitochondria in primary neurons. Additionally, PHB2 depletion exacerbated neuronal cell death induced by PrP106-126, whereas the overexpression of PHB2 alleviated PrP106-126 neuronal toxicity. Taken together, this study demonstrated that PHB2 is indispensable for PINK1/Parkin-mediated mitophagy in PrP106-126-treated neurons and protects neurons against the neurotoxicity of the prion peptide.
Keyword ：
mitophagy mitophagy neuronal death neuronal death PHB2 PHB2 PINK1/Parkin PINK1/Parkin prion disease prion disease prion peptide prion peptide PrP106-126 PrP106-126
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| GB/T 7714 | Zheng, Xiaohui , Liu, Kun , Xie, Qingqing et al. PHB2 Alleviates Neurotoxicity of Prion Peptide PrP106-126 via PINK1/Parkin-Dependent Mitophagy [J]. | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES , 2023 , 24 (21) . |
| MLA | Zheng, Xiaohui et al. "PHB2 Alleviates Neurotoxicity of Prion Peptide PrP106-126 via PINK1/Parkin-Dependent Mitophagy" . | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 24 . 21 (2023) . |
| APA | Zheng, Xiaohui , Liu, Kun , Xie, Qingqing , Xin, Hangkuo , Chen, Wei , Lin, Shengyu et al. PHB2 Alleviates Neurotoxicity of Prion Peptide PrP106-126 via PINK1/Parkin-Dependent Mitophagy . | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES , 2023 , 24 (21) . |
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神经退行性疾病是一种以神经元发生进行性变性和坏死为基础的中枢神经系性疾病，其普遍特征是错误折叠蛋白质的积累和线粒体损伤。线粒体作为细胞能量产出的中心，是神经元的主要能量来源，对维持神经元的结构和功能至关重要。受损的线粒体导致细胞中的三磷酸腺苷(ATP)供给不足和氧化应激损伤，甚至引起细胞死亡。线粒体自噬是细胞通过自噬-溶酶体途径选择性地清除衰老或受损线粒体的过程，是线粒体质量控制机制的重要组成部分，在维持细胞稳态方面发挥重要的作用。诸多研究表明，线粒体自噬与神经退行性疾病的发生和发展密不可分，激活线粒体自噬或改善线粒体自噬异常能在一定程度上缓解错误折叠蛋白积聚导致的神经损伤。笔者就线粒体自噬的发生机制、线粒体自噬的调控及其在神经退行性疾病发生发展中的作用进行综述，以期为神经退行性疾病的研究和治疗提供参考。
Keyword ：
神经损伤 神经损伤 神经退行性疾病 神经退行性疾病 线粒体 线粒体 线粒体自噬 线粒体自噬
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| GB/T 7714 | 郑小惠 , 刘坤 , 辛航阔 et al. 线粒体自噬在神经退行性疾病中调控的研究进展 [J]. | 中国畜牧兽医 , 2023 , 50 (02) : 490-499 . |
| MLA | 郑小惠 et al. "线粒体自噬在神经退行性疾病中调控的研究进展" . | 中国畜牧兽医 50 . 02 (2023) : 490-499 . |
| APA | 郑小惠 , 刘坤 , 辛航阔 , 谢青青 , 朱婷 . 线粒体自噬在神经退行性疾病中调控的研究进展 . | 中国畜牧兽医 , 2023 , 50 (02) , 490-499 . |
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为探究热休克蛋白70家族成员葡萄糖调节蛋白78(GRP78)在伪狂犬病病毒（PRV）感染宿主过程中的作用，本研究利用PRV Min-A株感染PK-15细胞后，采用western blot和间接免疫荧光试验（IFA）分别检测GRP78蛋白的表达和亚细胞定位情况，western blot结果显示，与对照组相比，PRV感染组的GRP78蛋白水平未见明显变化；IFA结果显示，PRV感染组细胞膜上GRP78的荧光强度明显增加。进一步通过生物素标记细胞膜蛋白，然后利用磁珠分离细胞膜蛋白，经western blot检测结果显示，PRV感染组细胞膜中GRP78的蛋白水平显著高于未感染PRV的对照组（P<0.05）。可见PRV感染不影响细胞内GRP78蛋白的表达，但能够增加GRP78在细胞膜中的含量。利用GRP78多克隆抗体（pAb）与PK-15细胞共孵育1 h后感染PRV，经空斑试验检测细胞上清中的病毒滴度，结果显示，与对照组相比，GRP78 p Ab孵育组的病毒滴度未见明显变化，表明GRP78并不是PRV侵入宿主细胞的关键因子。通过IFA检测GRP78与PRV在细胞中的共定位情况，结果显示，GRP78与PRV在细胞膜上无共定位，但二者在细胞质中有共定位。分别在PK-15细胞中下调或者过表达GRP78后感染PRV，采用western blot和空斑试验分别检测细胞内PRV的蛋白合成和细胞上清中的病毒滴度，结果显示，GRP78下调后PRV蛋白和病毒滴度显著低于对照组（P<0.05），而过表达GRP78组的病毒滴度明显高于对照组，表明GRP78参与调节PRV的释放。本研究首次表明GRP78作为一种重要的宿主因子参与调节PRV从宿主细胞内的释放，该结果为进一步研究PRV与宿主细胞的相互作用提供了参考依据。
Keyword ：
伪狂犬病病毒 伪狂犬病病毒 感染 感染 细胞膜 细胞膜 葡萄糖调节蛋白78 葡萄糖调节蛋白78
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| GB/T 7714 | 薛晓暖 , 郑小惠 , 辛航阔 et al. GRP78在伪狂犬病病毒感染过程中的作用研究 [J]. | 中国预防兽医学报 , 2022 , 44 (10) : 1039-1044,1058 . |
| MLA | 薛晓暖 et al. "GRP78在伪狂犬病病毒感染过程中的作用研究" . | 中国预防兽医学报 44 . 10 (2022) : 1039-1044,1058 . |
| APA | 薛晓暖 , 郑小惠 , 辛航阔 , 谢青青 , 刘坤 , 祁保民 et al. GRP78在伪狂犬病病毒感染过程中的作用研究 . | 中国预防兽医学报 , 2022 , 44 (10) , 1039-1044,1058 . |
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Viruses have evolved multiple strategies to manipulate their host's translational machinery for the synthesis of viral proteins. A common viral target is the alpha subunit of eukaryotic initiation factor 2 (eIF2 alpha). In this study, we show that global protein synthesis was increased but the eIF2 alpha phosphorylation level was markedly decreased in porcine kidney 15 (PK15) cells infected with pseudorabies virus (PRV), a swine herpesvirus. An increase in the eIF2 alpha phosphorylation level by salubrinal treatment or transfection of constructs expressing wild-type eIF2 alpha or an eIF2 alpha phosphomimetic [eIF2 alpha(S51D)] attenuated global protein synthesis and suppressed PRV replication. To explore the mechanism involved in the inhibition of eIF2 alpha phosphorylation during PRV infection, we examined the phosphorylation status of protein kinase R-like endoplasmic reticulum kinase (PERK) and double-stranded RNA-dependent protein kinase R (PKR), two kinases that regulate eIF2 alpha phosphorylation during infection with numerous viruses. We found that the level of neither phosphorylated (p)-PERK nor p-PKR was altered in PRV-infected cells or the lungs of infected mice. However, the expression of growth arrest and DNA damage-inducible protein 34 (GADD34), which promotes eIF2 alpha dephosphorylation by recruiting protein phosphatase 1 (PP1), was significantly induced both in vivo and in vitro. Knockdown of GADD34 and inhibition of PP1 activity by okadaic acid treatment led to increased eIF2 alpha phosphorylation but significantly suppressed global protein synthesis and inhibited PRV replication. Collectively, these results demonstrated that PRV induces GADD34 expression to promote eIF2 alpha dephosphorylation, thereby maintaining de novo protein synthesis and facilitating viral replication.
Keyword ：
eIF2 alpha eIF2 alpha GADD34 GADD34 host translation host translation phosphorylation phosphorylation PRV PRV
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| GB/T 7714 | Zhu, Ting , Jiang, Xueli , Xin, Hangkuo et al. GADD34-mediated dephosphorylation of eIF2α facilitates pseudorabies virus replication by maintaining de novo protein synthesis [J]. | VETERINARY RESEARCH , 2021 , 52 (1) . |
| MLA | Zhu, Ting et al. "GADD34-mediated dephosphorylation of eIF2α facilitates pseudorabies virus replication by maintaining de novo protein synthesis" . | VETERINARY RESEARCH 52 . 1 (2021) . |
| APA | Zhu, Ting , Jiang, Xueli , Xin, Hangkuo , Zheng, Xiaohui , Xue, Xiaonuan , Chen, Ji-Long et al. GADD34-mediated dephosphorylation of eIF2α facilitates pseudorabies virus replication by maintaining de novo protein synthesis . | VETERINARY RESEARCH , 2021 , 52 (1) . |
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本研究从疑似暴发伪狂犬病(pseudorabies,PR)猪场死亡的仔猪中分离到1株病毒,经组织病理学剖检、PCR试验、Western-blot分析,将其鉴定为伪狂犬病病毒(pseudorabies virus,PRV),遂被命名为SX-2018,该病毒在MDCK细胞上具有较强的适应性。将SX-2018株gE基因的序列与国内外21株PRV参考序列进行同源性比较,结果显示,SX-2018株gE基因与国内外参考毒株的核苷酸序列同源性为97.1%～99.9%,氨基酸同源性为94.5%～100%。遗传进化分析显示,SX-2012株与我国近几年新分离的猪伪狂犬变异毒株处于同一分支;并且与经典株相比,gE蛋白在第48位和第492位各有1个天冬氨酸的插入。为了进一步探讨SX-2018毒株的致病性,将SX-2018株和Min-A株分别感染BALB/c小鼠,分析两组小鼠的发病特征和组织损伤情况;结果显示,SX-2018组的小鼠死亡时间早于Min-A组;并且Min-A组小鼠的肺脏主要表现为间质性肺炎,而SX-2018组的小鼠以出血性肺炎为主。荧光定量PCR分别检测两组小鼠肺脏内PRV gI基因和炎症相关因子的变化;结果显示,SX-2018组小鼠肺组织内的gI基因以及炎症相关因子的mRNA水平显著地高于Min-A组小鼠。以上结果表明,本文成功分离到1株PRV变异毒株,并且与PRV经典株Min-A株相比,新分离的变异毒株可以诱导不同程度的病理损伤和炎症反应。
Keyword ：
gE gE 伪狂犬病病毒 伪狂犬病病毒 分离鉴定 分离鉴定 变异株 变异株 致病性分析 致病性分析
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| GB/T 7714 | 薛晓暖 , 姜雪梨 , 陈吉龙 et al. 猪伪狂犬病病毒SX-2018变异株的分离鉴定及其致病性研究 [J]. | 中国兽医科学 , 2021 , 51 (07) : 857-864 . |
| MLA | 薛晓暖 et al. "猪伪狂犬病病毒SX-2018变异株的分离鉴定及其致病性研究" . | 中国兽医科学 51 . 07 (2021) : 857-864 . |
| APA | 薛晓暖 , 姜雪梨 , 陈吉龙 , 祁保民 , 辛航阔 , 郑小惠 et al. 猪伪狂犬病病毒SX-2018变异株的分离鉴定及其致病性研究 . | 中国兽医科学 , 2021 , 51 (07) , 857-864 . |
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