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大肠杆菌多酚氧化酶的分子克隆及异源高效表达

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中国饲料 2024年第5期 26-31页

作者：邓卉余丹邹成义范景胜李斌屈东倪青松郑钰嘉陈瑾四川省畜牧科学研究院动物遗传育种四川省重点实验室四川成都610066

菜籽粕、棉籽粕等非粮饲料资源存在的酚类化合物芥子碱、棉酚等抗营养因子,严重影响了其饲喂价值。本课题组筛选出的一株大肠杆菌属芥子碱降解菌SDB2已被证实能通过分泌多酚氧化酶来降解芥子碱和棉酚,本试验旨在运用基因工程技术克隆SDB... 详细信息

菜籽粕、棉籽粕等非粮饲料资源存在的酚类化合物芥子碱、棉酚等抗营养因子,严重影响了其饲喂价值。本课题组筛选出的一株大肠杆菌属芥子碱降解菌SDB2已被证实能通过分泌多酚氧化酶来降解芥子碱和棉酚,本试验旨在运用基因工程技术克隆SDB2多酚氧化酶基因,实现其高效表达,为SDB2多酚氧化酶在饲料工业上的规模化应用奠定基础。从大肠杆菌SDB2中克隆出多酚氧化酶基因,将其与质粒pET28a连接后转化至大肠杆菌BL21感受态细胞中培养,通过PCR鉴定及质粒酶切验证方法构建重组菌株,同时对重组克隆基因进行生物信息学分析。采用“2×2”交叉试验设计,以温度和诱导剂IPTG浓度两个影响因素诱导SDB2多酚氧化酶基因在重组菌株中高效表达,经SDS-PAGE和WB双重验证后,确立最佳诱导条件,最终纯化出目标酶蛋白。结果表明:(1)成功克隆SDB2多酚氧化酶基因,构建出异源表达重组菌株。(2)克隆出的SDB2重组多酚氧化酶基因含目标核苷酸741个,总编码氨基酸263个(含标签氨基酸20个),氨基酸组成的蛋白相对分子质量为28499.0,理论等电点为6.94,据此预测出SDB2重组多酚氧化酶三维结构模型。(3)确立SDB2多酚氧化酶异源高效表达的最佳诱导条件为温度37℃,IPTG浓度1 mmol,该条件下表达出大量可溶性酶蛋白,有利于工业化生产。本试验条件下,大肠杆菌SDB2多酚氧化酶成功实现了分子克隆及异源高效表达,为我国非粮饲料资源实现高效利用提供了技术参考。

关键词：大肠杆菌多酚氧化酶克隆高效表达

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Ginsenoside Rg1 protects against ischemia-induced neuron damage by regulating the rno-miRNA-27a-3p/PPARγaxis

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BIOCELL 2023年第7期47卷 1583-1594页

作者： YUE GUAN TINGTING ZHANG JIANAN YU JIAWEI LIU WENYUAN LI yujia zheng JIALE WANG YUE LIU FENGGUO ZHAI Department of Clinical Medicine Heilongjiang Nursing CollegeHarbin150001China Department of Neurology Hongqi Hospital of Mudanjiang Medical UniversityMudanjiang157011China Department of Pharmacology Mudanjiang Medical UniversityMudanjiang157011China Department of Organic Chemistry Mudanjiang Medical UniversityMudanjiang157011China Department of Anatomy Mudanjiang Medical UniversityMudanjiang157011China Department of Rehabilitation Science Kobe University Graduate School of Health SciencesKobe654-0142Japan Institute of Natural Medicine Mudanjiang Medical UniversityMudanjiang157011China

A preliminary miRNA screening showed that expression levels of rno-miRNA-27a-3p were significantly increased in the serum and brain tissues of rats undergoing cerebral *** recent years,there is evidence of the protective capacity of the saponins extracted from panax ginseng and its primary active ingredient ginsenosideRg1oncerebral ischemic ***:Fetal rat neurons(FRNs)were cultured in glucose-and-serumfree medium and exposed to hypoxia to establish a cerebral ischemia model in vitro(oxygen and glucose deprivation model,OGD).Antioxidant indexes(CAT,SOD),inflammatory markers(MPO,TNF-αand IL-6),and the expression of apoptosis and proliferation associated proteins(NF kB-p65,Caspase 3-cleaved,BCL-2)were ***:Pre-treatment of Rg1(30–100μg/mL)could effectively inhibit the decline of antioxidant indexes(CAT,SOD)and increase in inflammatory markers(MPO,TNF-αand IL-6),and effectively inhibited the apoptosis in FRNs induced by OGD in a gradient-dependent *** mechanism analysis showed that the role of Rg1 in protecting against ischemia-induced neuron damage depends on its indirect up-regulation of PPAR protein via suppression of ***,these effects of Rg1 could be reversed by exogenous rno-miRNA-27a-3p and PPAR gene silencing in FRNs exposed to ***:To summarize,our study demonstrates that Rg1 could effectively attenuate neuronal damage caused by cerebral ischemia via the rno-miRNA-27a-3p/PPARγ***,clarification of the novel mechanism will certainly improve our previous understanding of the role of Rg1 and enhancing its level in treatments for alleviating ischemic brain injury.

关键词： Ginsenoside Rg1 rno-miRNA-27a-3p PPARγ Cerebral ischemia Neuron OGD

Functional characterization of Bmcap in uric acid metabolism in the silkworm

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Insect Science 2024年第1期31卷 147-156页

作者： Linmeng Tang Dehong Yang Zhiwei Liu Yaohui Wang Xu Yang yujia Liu Dongbin Chen zheng Tang Yongping Huang Key Laboratory of Insect Developmental and Evolutionary Biology Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghaiChina CAS Center for Excellence in Biotic Interactions University of Chinese Academy of SciencesBeijingChina Central Research Institute Shanghai Pharmaceuticals Holding Co.Ltd.ShanghaiChina Departments of Neonatology International Peace Maternity and Child Health Hospital of China Welfare InstitutionSchool of MedicineShanghai Jiao Tong UniversityShanghaiChina

After a millennium of domestication,numerous silkworm mutants have emerged that exhibit transparent epidermis,which is caused by abnormally low levels of uric *** identified the Bombyx mori gene Bmcap(BMSK0003832)as the homolog of cappuccino,a subunit of the biogenesis of lysosome-related organelles complex-1(BLOC-1)that has been extensively characterized in human,mouse,and insect species,by analyzing the amino acid sequences of putative purine metabolism *** the clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9 system,we disrupted Bmcap,resulting in decreased uric acid levels in the silkworm epidermis and a translucent skin *** the Bmcap mutant,the purine metabolism,nitrogen metabolism,pyrimidine metabolism,and membrane system were altered compared to the wild *** of lysosome-related organelle complex genes play a role in the pigmentation and biogenesis of lysosome-related organelles(LROs)in platelets,melanocytes,and *** exhibit unique morphologies and functions in various tissues and *** of the Bmcap mutant will enhance our understanding of the uric acid metabolic pathway in silkworms,and this mutant offers a valuable silkworm model for LRO studies.

关键词： BLOC-1 subunit Bombyx mori cappuccino translucent mutation uric acid

Low-frequency Stimulation at the Subiculum Prevents Extensive Secondary Epileptogenesis in Temporal Lobe Epilepsy

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Neuroscience Bulletin 2024年第5期40卷 564-576页

作者： yujia Shen Yiwei Gong Xiaoli Da Shajing Gao Shuo Zhang Minjuan Sun Yuanzhi Yang Xiaoyun Qiu Menghan Li Yang zheng Fan Fei Yi Wang Zhong Chen Cenglin Xu Key Laboratory of Neuropharmacology and Translational Medicine of Zhejiang Province School of Pharmaceutical ScienceZhejiang Chinese Medical UniversityHangzhou310053China Institute of Pharmacology and Toxicology NHC and CAMS Key Laboratory of Medical NeurobiologyCollege of Pharmaceutical SciencesZhejiang UniversityHangzhou310058China Department of Neurology First Affiliated Hospital of Zhejiang Chinese Medical UniversityHangzhou310003China Department of Pharmacy The First Affiliated Hospital of Zhejiang Chinese Medical UniversityHangzhou310003China

Secondary epileptogenesis is characterized by increased epileptic susceptibility and a tendency to generate epileptiform activities outside the primary *** is one of the major resultants of pharmacoresistance and failure of surgical outcomes in epilepsy,but still lacks effective ***,we aimed to test the effects of low-frequency stimulation(LFS)at the subiculum for secondary epileptogenesis in a mouse ***,secondary epileptogenesis was simulated at regions both contralateral and ipsilateral to the primary focus by applying successive kindling *** kindled at the right CA3 showed higher seizure susceptibilities at both the contralateral CA3 and the ipsilateral entorhinal cortex and had accelerated kindling processes compared with naive *** at the ipsilateral subiculum during the primary kindling progress at the right CA3 effectively prevented secondary epileptogenesis at both the contralateral CA3 and the ipsilateral entorhinal cortex,characterized by decreased seizure susceptibilities and a retarded kindling process at those secondary *** application along with the primary epileptogenesis was ***,the effects of LFS on secondary epileptogenesis were associated with its inhibitory effect at the secondary focus through interfering with the enhancement of synaptic connections between the primary and secondary *** results imply that LFS at the subiculum is an effective preventive strategy for extensive secondary epileptogenesis in temporal lobe epilepsy and present the subiculum as a target with potential translational importance.

关键词： Secondary epileptogenesis Low-frequency stimulation Subiculum Temporal lobe epilepsy

PIF4 interacts with ABI4 to serve as a transcriptional activator complex to promote seed dormancy by enhancing ABA biosynthesis and signaling

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Journal of Integrative Plant Biology 2024年第5期66卷 909-927页

作者： Xiaofeng Luo yujia Dai Baoshan Xian Jiahui Xu Ranran Zhang Muhammad Saad Rehmani Chuan zheng Xiaoting Zhao Kaitao Mao Xiaotong Ren Shaowei Wei Lei Wang Juan He Weiming Tan Junbo Du Weiguo Liu Shu Yuan Kai Shu Shaanxi Key Laboratory of Qinling Ecological Intelligent Monitoring and Protection School of Ecology and EnvironmentNorthwestern Polytechnical UniversityXi'an 710129China Research&Development Institute of Northwestern Polytechnical University in ShenzhenShenzhen 518057China College of Agronomy and Biotechnology China Agricultural UniversityBeijing 100193China Institute of Ecological Agriculture Sichuan Agricultural UniversityChengdu 611130China College of Resources Sichuan Agricultural UniversityChengdu 611130China

Transcriptional regulation plays a key role in the control of seed dormancy,and many transcription factors(TFs)have been ***,the mechanisms underlying the interactions between different TFs within a transcriptional complex regulating seed dormancy remain largely ***,we showed that TF PHYTOCHROME-INTERACTING FACTOR4(PIF4)physically interacted with the abscisic acid(ABA)signaling responsive TF ABSCISIC ACID INSENSITIVE4(ABI4)to act as a transcriptional complex to promote ABA biosynthesis and signaling,finally deepening primary seed *** pif4 and abi4 single mutants exhibited a decreased primary seed dormancy phenotype,with a synergistic effect in the pif4/abi4 double ***4 binds to ABI4 to form a heterodimer,and ABI4 stabilizes PIF4 at the protein level,whereas PIF4 does not affect the protein stabilization of ***,both TFs independently and synergistically promoted the expression of ABI4 and NCED6,a key gene for ABA *** genetic evidence is also consistent with the phenotypic,physiological and biochemical analysis ***,this study revealed a transcriptional regulatory cascade in which the PIF4–ABI4 transcriptional activator complex synergistically enhanced seed dormancy by facilitating ABA biosynthesis and signaling.

关键词： ABA ABI4 PIF4 seed dormancy and germination transcriptional complex

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Multi-omics joint analysis revealed the metabolic profile of retroperitoneal liposarcoma

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Frontiers of Medicine 2024年第2期18卷 375-393页

作者： Fu’an Xie yujia Niu Lanlan Lian Yue Wang Aobo Zhuang Guangting Yan Yantao Ren Xiaobing Chen Mengmeng Xiao Xi Li Zhe Xi Gen Zhang Dongmei Qin Kunrong Yang Zhigang zheng Quan Zhang Xiaogang Xia Peng Li Lingwei Gu Ting Wu Chenghua Luo Shu-Hai Lin Wengang Li Cancer Research Center Xiang’an Hospital of Xiamen UniversitySchool of MedicineXiamen UniversityXiamen361102China State Key Laboratory of Cellular Stress Biology School of Life SciencesFaculty of Medicine and Life SciencesXiamen UniversityXiamen361102China Xiamen University Research Center of Retroperitoneal Tumor Committee of Oncology Society of Chinese Medical Association Xiamen UniversityXiamen361102China Department of Laboratory Medicine Xiang’an Hospital of Xiamen UniversitySchool of MedicineXiamen UniversityXiamen361102China Department of Retroperitoneal Tumor Surgery Peking University International HospitalBeijing102206China School of Public Health Harvard UniversityBostonMA02115USA Laboratory of Biochemistry and Molecular Biology Research Department of Clinical LaboratoryFujian Medical University Cancer HospitalFuzhou350014China Surgery Department Affiliated Fuzhou First Hospital of Fujian Medical UniversityFuzhou350009China National Institute for Data Science in Health and Medicine Xiamen UniversityXiamen361102China Department of Hepatobiliary Surgery Xiang’an Hospital of Xiamen UniversitySchool of MedicineXiamen UniversityXiamen361102China

Retroperitoneal liposarcoma(RLPS)is the main subtype of retroperitoneal soft sarcoma(RSTS)and has a poor prognosis and few treatment options,except for *** proteomic and metabolic profiles of RLPS have remained *** aim of our study was to reveal the metabolic profile of ***,we performed proteomic analysis(n=10),metabolomic analysis(n=51),and lipidomic analysis(n=50)of retroperitoneal dedifferentiated liposarcoma(RDDLPS)and retroperitoneal well-differentiated liposarcoma(RWDLPS)tissue and paired adjacent adipose tissue obtained during *** analysis mainly revealed that glycolysis,purine metabolism,pyrimidine metabolism and phospholipid formation were upregulated in both RDDLPS and RWDLPS tissue compared with the adjacent adipose tissue,whereas the tricarboxylic acid(TCA)cycle,lipid absorption and synthesis,fatty acid degradation and biosynthesis,as well as glycine,serine,and threonine metabolism were *** particular importance,the glycolytic inhibitor 2-deoxy-D-glucose and pentose phosphate pathway(PPP)inhibitor RRX-001 significantly promoted the antitumor effects of the MDM2 inhibitor RG7112 and CDK4 inhibitor *** study not only describes the metabolic profiles of RDDLPS and RWDLPS,but also offers potential therapeutic targets and strategies for RLPS.

关键词： RLPS proteomics metabolomics lipidomics metabolism

TNF-α-induced Tim-3 expression marks the dysfunction of infiltrating natural killer cells in human esophageal cancer

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TNF-α-induced Tim-3 expression marks the dysfunction of inf...

第十三届全国免疫学学术大会

作者： yujia zheng Yi Zhang Lan Huang Biotherapy Center the First Affiliated Hospital of Zhengzhou University

Objective: Impairment of natural killer(NK) cell activity is an important mechanism of tumor immunoevasion. T-cell immunoglobulin domain and mucin domain-3(Tim-3) is an activation-induced inhibitory molecule, inducing effector lymphocyte exhaustion in chronic viral infection and cancers. However, its function in NK cells in human esophageal cancer remains ***: We prospectively collected peripheral blood and tumor samples from 53 patients with esophageal cancer. Peripheral and tumor-infiltrating NK cells were analyzed for Tim-3, Annexin V, CD69, CD107 a and IFN-g expression by flow cytometry. Quantitative real-time PCR was used to test relative mRNA expression of IFN-g, granzyme B, perforin and NKG2 D in sorted Tim-3+ NK cells and Tim-3-NK cells, respectively. NK cells isolated from healthy donors were treated with recombinant TNF-α to induce Tim-3 expression. Tim-3 and TNF-α mRNA levels in tumor tissues were measured in both humans and mice. Finally, associations between NK cell frequencies with pathological parameters were ***: We observed up-regulation of Tim-3 expression on NK cells from esophageal cancer patients, especially at the tumor site. Furthermore, tumor-infiltrating NK cells with high Tim-3 expression exhibited a phenotype with enhanced dysfunction. In vitro, Tim-3 expression on NK cells isolated from blood of healthy donors can be induced by recombinant TNF-α via NF-κB pathway. In both animal models and patients, the Tim-3 level was positively correlated with TNF-α expression in esophageal cancer tissues. Finally, higher Tim-3 level on tumor-infiltrating NK cells is correlated with big tumor size, nodal status and poor stage in patients with esophageal ***: Taken together, Tim-3 may play a crucial role to induce NK cell dysfunction in tumor microenvironmentand could serve as a potential biomarker for prognosis of esophageal cancer.

关键词： Tumor microenvironment NK cells Tim-3 TNF-α esophageal cancer

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结合自训练模型的命名实体识别方法

山东大学学报（工学版） 2024年第2期54卷 96-102页

作者：肖伟郑更生陈钰佳武汉工程大学计算机科学与工程学院、人工智能学院湖北武汉430205 智能机器人湖北省重点实验室湖北武汉430205

针对命名实体识别数据集中存在某些实体类别样本过少,使模型学习该类别特征能力较差,导致整体性能较低的问题,提出结合自训练模型的命名实体识别方法。利用已有的命名实体识别数据集训练一个教师模型,通过改进的文本相似度函数搜寻与原... 详细信息

针对命名实体识别数据集中存在某些实体类别样本过少,使模型学习该类别特征能力较差,导致整体性能较低的问题,提出结合自训练模型的命名实体识别方法。利用已有的命名实体识别数据集训练一个教师模型,通过改进的文本相似度函数搜寻与原数据集最相似的无标签文本,利用教师模型对无标签文本生成伪标签,并将伪标签与有标签数据集混合重新训练一个学生模型用于下游的命名实体识别任务。试验结果表明,相较基线模型,该方法在公共数据集MSRA、CONLL03和法律实体识别数据集上取得更优的性能。

关键词：命名实体识别自训练文本相似度自然语言处理少样本

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均相与多相催化剂在CO_(2)环加成反应中的研究进展

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燃料化学学报（中英文） 2024年第1期52卷 105-130页

作者：李宁宁刘轩博张宇航王雨佳常涛朱正河北工程大学材料科学与工程学院邯郸市CO_(2)催化转化重点实验室河北邯郸056038 河北省杂环化合物重点实验室河北邯郸056005

二氧化碳(CO_(2))是一种主要的人为温室气体,主要由化学、热电和钢铁工业以及运输等部门产生。大气层中CO_(2)浓度的增加是导致诸多环境问题的主要原因,如全球变暖、海平面上升和全球气温升高。然而,CO_(2)作为一种可再生、廉价和无毒... 详细信息

二氧化碳(CO_(2))是一种主要的人为温室气体,主要由化学、热电和钢铁工业以及运输等部门产生。大气层中CO_(2)浓度的增加是导致诸多环境问题的主要原因,如全球变暖、海平面上升和全球气温升高。然而,CO_(2)作为一种可再生、廉价和无毒的化学原料,可用来生产具有高附加值的化学品,进而降低碳浓度。五元环碳酸酯由于其优越的物理化学特性,如高沸点、高偶极矩和生物降解等性能而被广泛应用。由环氧化合物和CO_(2)合成环碳酸酯是迄今为止研究较多的方法。然而,由于CO_(2)的高热稳定性和动力学惰性,使其作为反应原料需要大量的能量投入,可能导致的结果是CO_(2)浓度是一个净增长过程。因此,利用CO_(2)作为C1构筑单元是一个长期的挑战。本工作基于CO_(2)固定反应机制,概述了各种类型的均相和多相催化剂在CO_(2)固定反应合成精细化学品环状碳酸酯中的研究进展,包括有机催化剂、离子液体、金属有机框架化合物、多孔有机聚合物等。目前,几乎所有类别催化剂均可以在室温和低压力下,以实验室规模成功地使用纯CO_(2)将其固定到末端环氧化合物上,对于非末端环氧化合物通过更高的温度和压力以实现相应转化。同时,分析了催化剂在多取代环氧化合物或生物衍生环氧化合物转化、低浓度CO_(2)转化和实现工业化三个方面所面临的挑战,并提出了未来相关研究努力的方向。

关键词： CO_(2) 环氧化合物催化剂环加成反应环碳酸酯

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