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The ubiquitin ligase Nedd4-2 mediates the regulation of PepT2 by mTORC1 in bovine mammary epithelial cells

Animal Nutrition 2022年第3期10卷 12-18页

作者： Caihong Wang Fengqi Zhao Jianxin Liu Hongyun liu College of Animal Science Zhejiang UniversityHangzhou310058China Department of Animal Science and Veterinary Sciences University of VermontBurlingtonVT05405USA

Peptide transporter 2(PepT2)transports short peptides from the blood into bovine mammary epithelial cells(BMEC)to stimulate milk protein synthesis.Despite the fact that the effect of PepT2 is acknowledged in BMEC,little is known about its regulation.This study was completed to investigate the role of mammalian target of the rapamycin(mTOR)signaling in regulating the expression and function of PepT2 in BMEC.The regulation of PepT2 by mTOR in BMEC was studied in vitro using peptide transport assay,gene silencing,Western blot.The membrane expression of PepT2 and the uptake of b-Ala-Lys-N-7-amino-4-methylcoumarin-3-acetic acid(b-Ala-Lys-AMCA),a model dipeptide,in BMEC were reduced by rapamycin(a mTOR inhibitor)and silencing of either mTOR complex 1(mTORC1)or mTOR complex 2(mTORC2),stimulated by DEP domain-containing mTOR-interacting protein(DEPTOR,endogenous inhibitor of mTORC1 and mTORC2)silencing.The trafficking of PepT2 to the membrane and the uptake of b-Ala-Lys-AMCA was promoted by neuronal precursor cell-expressed developmentally down-regulated 4 isoform 2(Nedd4-2)silencing.The effects of knockdown of mTORC1,but not mTORC2,on cell membrane expression and transport activity of PepT2 was abolished by Nedd4-2 silencing.With immunofluorescence staining,PepT2 was identified to be interacting with Nedd4-2.The Nedd4-2 expression and the interaction between PepT2 and Nedd4-2 was increased through mTORC1 knockdown,indicating an increased ubiquitination of PepT2.The results revealed that mTORC1 can regulate the expression and function of PepT2 through Nedd4-2 in BMEC.

关键词： mTORC1 mTORC2 Nedd4-2 PepT2 Peptide uptake BMEC

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Constraints on the utilization of cereal straw in lactating dairy cows:A review from the perspective of systems biology

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Animal Nutrition 2022年第2期9卷 240-248页

作者： Bing Wang Huizeng Sun Diming Wang Hongyun liu Jianxin Liu Institute of Dairy Science MoE Key Laboratory of Molecular Animal NutritionCollege of Animal SciencesZhejiang UniversityHangzhouChina State Key Laboratory of Animal Nutrition College of Animal Science and TechnologyChina Agricultural UniversityBeijingChina

Cereal straw,a human inedible crop byproduct,can be used as a roughage source in ruminants.However,the nutrition density and palatability are very low,limiting its efficient utilization in animal production.This review aims to systematically provide an overview of the limitations of cereal straws,which is crucial for developing new strategies to enhance the efficient use of cereal straws by lactating dairy cows.Evolutionary molecular biology makes it possible to comprehensively understand the limitations of using cereal straw as a roughage source in dairy cows by different techniques,e.g.,multi-omics.Main constraints for utilization of cereal straw and stover in lactating dairy cows include low contents of easily fermented carbohydrates(pectin)and essential amino acids(Met,Phe,and branched-chain amino acids),high content of lignin and silica,and low nutrient digestibility.These cause insufficient supply of the precursors for milk synthesis and result in increased loss of nutrients in feces and urine.Several molecular mechanisms are revealed by multi-omics techniques,including changed amino acid and glucose metabolism,altered rumen microbial composition and function,and differential expression of miRNAs,mRNA,and protein in multi-organs that are associated with milk synthesis.These can be targets of approaches to improve the utilization of cereal straw by dairy cows.In addition,much attention should be given to the efficient countermeasures,including pretreatments by fibrolytic enzymes or steam explosion,dietary formulations such as supplement of pectin,methionine,and branched-chain amino acids,and feeding with other functional feedstuffs,which may improve the feeding and economic value of cereal straw for lactating dairy cows.The newly revealed functional genes(such as BAG3 in the rumen,PC in the liver,CSN1S2 in the mammary gland)and biomarkers(hippuric acid)as well as the integrative signaling and metabolic pathways(phenylalanine metabolism)related to the shortages of cereal straws could be used as nutritional or

关键词： Amino acid metabolism Cereal straw Glucose utilization Metabolites Rumen microbiome

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Heat stress affects dairy cow health status through blood oxygen availability

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Journal of Animal Science and Biotechnology 2023年第6期14卷 2663-2673页

作者： Jia Zeng Jie Cai Diming Wang Hongyun liu Huizeng Sun Jianxin Liu Key Laboratory of Dairy Cow Genetic Improvement and Milk Quality Research of Zhejiang Province College of Animal SciencesZhejiang UniversityHangzhouChina Ministry of Education Key Laboratory of Molecular Animal Nutrition Zhejiang UniversityHangzhouChina

Background Rises in global warming and extreme weather occurrence make the risk of heat stress(HS)induced by high ambient temperatures more likely in high-yielding dairy cows,resulting in low milk quality and yield.In ani-mals,oxygen is involved in many physiological and metabolic processes,but the effects of HS on oxygen metabolism remain unclear.Thus,the current study aimed to investigate how oxygen metabolism plays a role in health status of dairy cows by measuring the milk yield,milk composition,and blood biochemical variables of cows under different levels of HS:none(No-HS),mild(Mild-HS),and moderate HS(Mod-HS).Results The HS significantly increased rectal temperature(Ptreat<0.01)and respiration rate(Ptreat<0.01).Under Mod-HS,greater Na+(P<0.05)and lower total CO_(2),and pH(P<0.05)were observed relative to those under No-HS and Mild-HS.Oxygen concentrations in both coccygeal artery and mammary vein(Ptreat<0.01)were lower under Mod-HS than under No-HS.Coccygeal vein concentrations of heat shock protein 90(HSP90)(P<0.05)increased during Mod-HS compared with those in cows under No-HS.Malondialdehyde increased during Mod-HS,and glu-tathione peroxidase(P<0.01)increased during Mild-HS.Coccygeal vein concentrations of vascular endothelial growth factor(P<0.01),heme oxygenase-1(P<0.01),and hypoxia-inducible factor 1α(P<0.01)were greater in cows under Mod-HS than those under No-HS.Red blood cell count(P<0.01)and hemoglobin concentration(P<0.01)were lower in the coccygeal vein of dairy cows under Mild-and Mod-HS than those of cows under No-HS.Conclusions Exposure to HS negatively impacts the health status and lactation performance of dairy cows by limit-ing oxygen metabolism and transportation.However,the specific mechanism by which HS affects mammary function in cows remains unclear and requires further exploration.

关键词： Dairy cow Health status Heat stress Lactation performance Oxygen metabolism

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Blood neutrophil extracellular traps:a novel target for the assessment of mammary health in transition dairy cows

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Journal of Animal Science and Biotechnology 2023年第3期14卷 1068-1079页

作者： Luyi Jiang Huizeng Sun Fengfei Gu Jin He Fengqi Zhao Jianxin Liu Institute of Dairy Science College of Animal SciencesZhejiang UniversityHangzhouChina Ministry of Education Key Laboratory of Molecular Animal Nutrition Zhejiang UniversityHangzhouChina Institute of Animal Genetics and Reproduction College of Animal SciencesZhejiang UniversityHangzhouChina Department of Animal&Veterinary Sciences University of VermontBurlingtonUSA

Background:Mammary health is important for transition dairy cows and has been well recognized to exert decisive effects on animal welfare.However,the factors influencing mammary health are still unclear.Differential somatic cell count(DSCC)could reflect the mastitis risk since it is the percentage of neutrophils plus lymphocytes in total somatic cells and could be reflective of mammary health of dairy cows.This work aimed to investigate the assessment and prognosis of the health of transition cows based on blood neutrophil extracellular traps(NETs).Results:Eighty-four transition Holstein dairy cows were selected.The serum was sampled in all the animals at week 1 pre-and postpartum,and milk was sampled at week 1 postpartum.Based on the DSCC in milk at week 1,cows with lower(7.4%±4.07%,n=15)and higher(83.3%±1.21%,n=15)DSCCs were selected.High DSCC cows had higher levels of red blood cell counts(P<0.05),hemoglobin(P=0.07),and hematocrit(P=0.05),higher concentrations of serum oxidative variables[reactive oxygen species(P<0.05),malondialdehyde(P<0.05),protein carbonyl(P<0.05),and 8-hydroxy-2-deoxyguanosine(P=0.07)],higher levels of serum and milk NETs(P<0.05)and blood-milk barrier indicators,including serumβ-casein(P=0.05)and milk immunoglobulin G2(P=0.09),than those of low DSCC cows.In addition,lower concentrations of serum nutrient metabolites(cholesterol and albumin)(P<0.05)and a lower level of serum deoxyribonuclease I(P=0.09)were observed in high DSCC cows than in low DSCC cows.Among the assessments performed using levels of the three prepartum serum parameters(NETs,deoxyribonuclease I andβ-casein),the area under the curve(0.973)of NETs was the highest.In addition,the sensitivity(1.00)and specificity(0.93)were observed for the discrimination of these cows using NETs levels with a critical value of 32.2 ng/mL(P<0.05).Conclusions:The formation of NETs in blood in transition dairy cows may damage the integrity of the blood-milk barrier and thereby increase the risk for mastitis in postpartum cows.

关键词： Blood-milk barrier Differential somatic cell count Mastitis risk Neutrophil extracellular traps

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Maximum Power Point Tracking With Fractional Order High Pass Filter for Proton Exchange Membrane Fuel Cell

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IEEE/CAA Journal of Automatica Sinica 2017年第1期4卷 70-79页

作者： Jianxin Liu Tiebiao Zhao YangQuan Chen School of Mechanical Engineering Xihua University University of California

Proton exchange membrane fuel cell(PEMFC) is widely recognized as a potentially renewable and green energy source based on hydrogen. Maximum power point tracking(MPPT) is one of the most important working conditions to be considered. In order to improve the performance such as convergence and robustness under disturbance and uncertainty,a fractional order high pass filter(FOHPF) is applied for the MPPT controller design based on the traditional extremum seeking control(ESC). The controller is designed with integerorder integrator(IO-I) and low pass filter(IO-LPF) together with fractional order high pass filter(FOHPF), by substituting the normal HPF in the original ESC system. With this FOHPF ESC,better convergence and smoother performance are achieved while maintaining the robust specifications. First, tracking stability is discussed under the commensurate-order condition. Then,simulation results are included to validate the proposed new FOHPF ESC scheme under disturbance. Finally, comparison results between FOHPF ESC and the traditional ESC method are also provided.

关键词： Extremum seeking control(ESC) fractional order high pass filter(FOHPF) fuel cell fractional controller stability maximum power point tracking(MPPT)

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Linear stability theory with the equivalent spanwise wavenumber correction in 3D boundary layers

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Applied Mathematics and Mechanics(English Edition) 2019年第3期40卷 407-420页

作者： Runjie SONG Shaolong ZHANG Jianxin Liu Laboratory for High-Speed Aerodynamics Tianjin University Institute of Fluid Physics China Academy of Engineering Physics

The prediction on small disturbance propagation in complex three-dimensional(3D) boundary layers is of great significance in transition prediction methodology, especially in the aircraft design. In this paper, the linear stability theory(LST) with the equivalent spanwise wavenumber correction(ESWC) is proposed in order to accurately predict the linear evolution of a disturbance in a kind of boundary layer flow with a vital variation in the spanwise direction. The LST with the ESWC takes not only the scale of the mean flow with the significant variation but also the wavenumber evolution of the disturbance itself. Compared with the conventional LST, the results obtained by the new method are in excellent agreement with those of the numerical simulations. The LST with the ESWC is an effective method on the prediction of the disturbance evolution in 3D boundary layers, which improves the prediction of the LST in the applications to complex 3D boundary layers greatly.

关键词： three-dimensional(3D)boundary layer prediction of disturbance propagation equivalent spanwise wavenumber linear stability theory(LST) numerical simulation

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Gut health benefit and application of postbiotics in animal production

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Journal of Animal Science and Biotechnology 2022年第4期13卷 999-1010页

作者： Yifan Zhong Shanshan Wang Hanqiu Di Zhaoxi Deng Jianxin Liu Haifeng Wang College of Animal Science Zhejiang UniversityThe Key Laboratory of Molecular Animal NutritionMinistry of EducationHangzhou 310058China

Gut homeostasis is of importance to host health and imbalance of the gut usually leads to disorders or diseases for both human and animal.Postbiotics have been applied in manipulating of gut health,and utilization of postbiotics threads new lights into the host health.Compared with the application of probiotics,the characteristics such as stability and safety of postbiotics make it a potential alternative to probiotics.Studies have reported the beneficial effects of components derived from postbiotics,mainly through the mechanisms including inhibition of pathogens,strengthen gut barrier,and/or regulation of immunity of the host.In this review,we summarized the characteristics of postbiotics,main compounds of postbiotics,potential mechanisms in gut health,and their application in animal production.

关键词： Animal production Gut health Postbiotics

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Improvement for expansion of parabolized stability equation method in boundary layer stability analysis

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Applied Mathematics and Mechanics(English Edition) 2018年第12期39卷 1737-1754页

作者： Yufeng HAN Jianxin Liu Jisheng LUO Department of Mechanics Tianjin UniversityTianjin 300072China Tianjin Key Laboratory of Modern Engineering Mechanics Tianjin 300072China

An improved expansion of the parabolized stability equation(iEPSE) method is proposed for the accurate linear instability prediction in boundary layers. It is a local eigenvalue problem, and the streamwise wavenumber α and its streamwise gradient dα/dx are unknown variables. This eigenvalue problem is solved for the eigenvalue dα/dx with an initial α, and the correction of α is performed with the conservation relation used in the PSE. The i EPSE is validated in several compressible and incompressible boundary layers. The computational results show that the prediction accuracy of the i EPSE is significantly higher than that of the ESPE, and it is in excellent agreement with the PSE which is regarded as the baseline for comparison. In addition, the unphysical multiple eigenmode problem in the EPSE is solved by using the i EPSE. As a local non-parallel stability analysis tool, the i EPSE has great potential application in the eNtransition prediction in general three-dimensional boundary layers.

关键词： parabolized stability equation(PSE) instability non-parallelism boundary layer

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Dietary supplementation with N-carbamoylglutamate initiated from the prepartum stage improves lactation performance of postpartum dairy cows

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Animal Nutrition 2021年第1期7卷 232-238页

作者： Fengfei Gu Chao Miao Luyi Jiang Diming Wang Hongyun liu Jianxin Liu Institute of Dairy Science MoE Key Laboratory of Molecular Animal NutritionCollege of Animal SciencesZhejiang UniversityHangzhou310058China

The objective of this study was to investigate the effects of supplementing N-carbamoylglutamate(NCG),an Arg enhancer,on amino acid(AA)supply and utilization and productive performance of early-lactating dairy cows.Thirty multiparous Chinese Holstein dairy cows were randomly divided into con-trol(CON,n=15)and NCG(CON diet supplemented with NCG at 20 g/d per cow,n=15)groups at 4 wk before calving.Diets were offered individually in tie-stalls,and NCG was supplemented by top-dress feeding onto total mixed ration for the NCG group.The experiment lasted until wk 10 after calving.Dry matter intake tended to be higher(P=0.06),and yields of milk(P<0.01),milk protein(P<0.01),and milk fat(P<0.01)were higher in the NCG-cows than in the CON-cows.Plasma activities of aspartate aminotransferase(P<0.01),alanine aminotransferase(P=0.03),and plasma level ofβ-hydroxybutyrate(P=0.04)were lower in the NCG-cows than in the CON-cows,whereas plasma glucose(P=0.05)and nitric oxide(NO,P<0.01)concentrations were higher.Coccygeal vein concentrations of Cys(P<0.01),Pro(P<0.01),Tyr(P=0.05),most essential AA except Thr and His(P<0.01),total essential AA(P<0.01),and total AA(P<0.01)were higher in the NCG-cows than in the CON-cows.The arterial supply of all AA was greater in the NCG-cows than in the CON-cows.The NCG-cows had higher mammary plasma flow of AA(P=0.04)and clearance rate of Cys(P<0.01),Pro(P<0.01)and Asp(P<0.01),and higher ratios of uptake to output of Met(P=0.05),Lys(P<0.01),Cys(P=0.01),Pro(P=0.03),and Asp(P=0.01).In summary,addition of NCG initiated from the prepartum period improved the lactation performance of postpartum dairy cows,which might attribute to greater Arg and NO concentrations,as well as improved AA supply and utilization,liver function,and feed intake in these cows.

关键词： N-carbamoylglutamate Arginine Transition cow Lactation performance

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Automatic discovery of stateful variables in network protocol software based on replay analysis

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Frontiers of Information Technology & Electronic Engineering 2023年第3期24卷 403-416页

作者： jianxin HUANG Bo YU Runhao liu Jinshu SU College of Computer Science and Technology National University of Defense TechnologyChangsha 410073China Academy of Military Science Beijing 100091China

Network protocol software is usually characterized by complicated functions and a vast state space.In this type of program,a massive number of stateful variables that are used to represent the evolution of the states and store some information about the sessions are prone to potentialﬂaws caused by violations of protocol specification requirements and program logic.Discovering such variables is significant in discovering and exploiting vulnerabilities in protocol software,and still needs massive manual verifications.In this paper,we propose a novel method that could automatically discover the use of stateful variables in network protocol software.The core idea is that a stateful variable features information of the communication entities and the software states,so it will exist in the form of a global or static variable during program execution.Based on recording and replaying a protocol program’s execution,varieties of variables in the life cycle can be tracked with the technique of dynamic instrument.We draw up some rules from multiple dimensions by taking full advantage of the existing vulnerability knowledge to determine whether the data stored in critical memory areas have stateful characteristics.We also implement a prototype system that can discover stateful variables automatically and then perform it on nine programs in Pro FuzzBench and two complex real-world software programs.With the help of available open-source code,the evaluation results show that the average true positive rate(TPR)can reach 82%and the average precision can be approximately up to 96%.

关键词： Stateful variables Network protocol software Program analysis technology Network security

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