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重组多杀性巴氏杆菌毒素介导FGFRL1诱导Hela细胞增殖研究

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农业生物技术学报 2024年第8期32卷 1899-1908页

作者：林忠森赵勤袁建林伍锐颜其贵文翼平黄小波杜森焱曹三杰四川农业大学动物医学院/猪病研究中心/国家级动物类实验教学示范中心/农业农村部兽用药物与兽医诊断技术四川科学观测站

成纤维细胞生长因子受体样因子1 (fibroblast growth factor receptor-like 1, FGFRL1)作为成纤维细胞生长因子受体家族成员，在调控细胞增殖、分化上起作用。多杀性巴氏杆菌毒素(Pasteurella multocida toxin, PMT)是多杀性巴氏杆菌(P.... 详细信息

成纤维细胞生长因子受体样因子1 (fibroblast growth factor receptor-like 1, FGFRL1)作为成纤维细胞生长因子受体家族成员，在调控细胞增殖、分化上起作用。多杀性巴氏杆菌毒素(Pasteurella multocida toxin, PMT)是多杀性巴氏杆菌(P. multocida, Pm)产生的重要毒力因子，属于一种促有丝分裂原蛋白，在多种细胞上引起细胞增殖，导致组织增生、占位，使其他正常组织细胞生存受限而死亡，但目前PMT引起促增殖作用的机制以及介导的因子均不清晰。前期CRISPR/Cas9全基因文库筛选发现FGFRL1提高了重组PMT (recombinant PMT, rPMT)蛋白染毒处理的Hela细胞存活率。为探究PMT在Hela细胞上的促增殖作用是否通过FGFRL1介导，本研究分别使用0、0.01、0.1、1.0μg/mL的rPMT蛋白处理正常及FGFRL1基因缺失的Hela细胞各48 h,CCK-8 (Cell Counting Kit-8)法检测细胞增殖率、Ed U-488标记法检测细胞DNA复制情况、划痕试验法检测增殖细胞的迁移率。结果显示，在rPMT浓度为0.1μg/mL处理48 h时，缺失FGFRL1的Hela细胞增殖显著下降(P<0.05)，而划痕恢复率与正常Hela细胞相比下降接近70%(P<0.05)。说明FGFRL1的缺失减弱了rPMT对Hela的促增殖作用。利用实时荧光定量PCR检测rPMT处理后Hela细胞磷脂酰肌醇3激酶/丝氨酸-苏氨酸蛋白激酶(phosphatidylinositol 3kinase/serine-threonine protein kinase, PI3K-AKT)通路上游因子变化水平、Western blot检测该通路关键激活因子AKT的磷酸化水平进行初步验证。结果显示：r PMT处理后PI3K-AKT通路上游因子转录水平显著升高(P<0.05)，缺失FGFRL1的Hela细胞AKT与磷酸化AKT水平极显著低于正常Hela细胞(P<0.01)，说明PI3K-AKT通路激活受到抑制。由此可以初步确认PMT可能通过FGFRL1靶向PI3K-AKT通路引起Hela细胞增殖。本研究丰富了PMT促增殖作用的理论基础，初步探索了PMT引起细胞增殖的通路激活方式，为进一步揭示PMT的促增殖作用提供参考。

关键词：成纤维细胞生长因子受体样因子1(FGFRL1) 重组多杀性巴氏杆菌毒素(rPMT) Hela细胞细胞增殖 PI3K-AKT通路

ArcA Gene Expression Level of Actinobacillus pleuropneumoniae in Anaerobic Culture

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畜牧兽医学报 2009年第S1期40卷 34-38页

作者： DUAN Li-li WEN Xin-tian cao san-jie HUANG Xiao-bo Laboratory of Animal Infectious Disease and Microarray Key Laboratory of Animal Disease and Human Health of Sichuan ProvinceCollege of Animal MedicineSichuan Agricultural University

In the present study,mRNA levels of the ArcA gene of Actinobacillus pleuropneumoniae serotype1 were measured during response to stress by growing under anaerobic *** levels were measured by semi-quantitative RT-PCR using the housekeeping gene recF as an internal *** expression of ArcA was undetectable until about 3 hours under standard culture conditions,but it was promptly and highly expressed in anaerobic *** results are consistent with ArcA being a potential virulence gene of Actinobacillus pleuropneumoniae,and likely playing an important role in pathogenesis caused by this organism.

关键词： semi-quantitative RT-PCR Actinobacillus pleuropneumoniae ArcA gene anaerobic stress

低浓度猪链球菌溶血素诱导猪肾细胞凋亡方式研究

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农业生物技术学报 2024年第2期32卷 355-366页

作者：戴世馨赵勤徐魁牟欣然杜森焱伍锐文翼平黄小波颜其贵曹三杰四川农业大学动物医学院/猪病研究中心/国家级动物类实验教学示范中心/农业农村部兽用药物与兽医诊断技术四川科学观测站

猪链球菌(Streptococcus suis)是一种危害严重的人畜共患病原菌，猪链球菌溶血素(suilysin, Sly)在猪链球菌的侵袭和致病中起关键作用，对多种类型的细胞具有毒性作用，但目前关于Sly的细胞毒性机制研究较少。为了探讨低浓度(0.3～0.6μ... 详细信息

猪链球菌(Streptococcus suis)是一种危害严重的人畜共患病原菌，猪链球菌溶血素(suilysin, Sly)在猪链球菌的侵袭和致病中起关键作用，对多种类型的细胞具有毒性作用，但目前关于Sly的细胞毒性机制研究较少。为了探讨低浓度(0.3～0.6μg/mL) Sly诱导猪肾细胞PK-15(porcine kidney-15)的死亡方式，本研究首先采用原核表达并纯化获得重组猪链球菌溶血素(recombinant suilysin, rSly)，通过乳酸脱氢酶(lactate dehydrogenase, LDH)释放检测rSly对PK-15细胞的毒性作用，并针对4种常见程序性细胞死亡方式，利用qPCR、间接免疫荧光、Werstern blot和流式细胞术对rSly作用于PK-15细胞12及24 h诱导的细胞死亡方式进行验证。结果显示，成功表达具有较高溶血活性(溶血单位为214)的rSly毒素蛋白，确定其对PK-15细胞的毒性作用呈浓度依赖性，并且发现rSly作用PK-15细胞12和24 h均引起PK-15细胞凋亡相关基因—半胱氨酸天冬氨酸蛋白酶3(cysteine aspartic acid specific protease 3, Caspase 3)、Caspase 9的转录水平显著升高(P<0.05)，细胞凋亡率与r Sly浓度呈正相关，0.6μg/mL rSly作用PK-15细胞24 h，细胞凋亡率从9.35%增至49.9%；自噬关键基因—微管相关蛋白轻链3B(microtubule associated protein 1 light chain 3β, LC3β)基因转录水平无明显变化且无自噬小体形成；焦亡关键基因—胃泌素D(gasdermin D,GSDMD)的转录水平无明显变化且GSDMD蛋白未发生明显切割；坏死性凋亡相关基因下调，且磷酸化混合系列蛋白激酶样结构域蛋白(phosphorylated mixed lineage kinase domain-like protein, p-MLKL)表达水平受到抑制。上述结果提示低浓度rSly诱导PK-15细胞死亡的方式主要是凋亡。本研究丰富了Sly毒性作用的细胞模型，有助于揭示参加猪链球菌感染发展的分子机制。

关键词：重组猪链球菌溶血素(rSly) 猪肾细胞(PK-15) 细胞毒性细胞死亡方式细胞凋亡

Identification and Detection of Actinobacillus pleuropneumoniae in Infected and Subclinically Infected Pigs by Multiplex PCR Based on the Genes ApxIVA and OmlA

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Agricultural Sciences in China 2006年第2期5卷 146-154页

作者： XIAO Guo-sheng cao san-jie DUAN Li-li WEN Xin-tian MA Xiao-ping CHEN Hua-mei Laboratory of Animal Infectious Disease and Microarray/Key Laboratory of Animal Disease and Human Health of Sichuan Province College of Animal Science and Veterinary Medicine Sichuan Agricultural University Ya＇an 625014 P.R.China

PCRs based on different genes of Actinobacillus pleuropneumoniae have been developed for detecting and identifying A. pleuropneumoniae. Some of them could amplify positive fragments from the phylogenetically closely related species bacteria. To improve veracity and specificity of PCR, a species-specific multiplex PCR assay was developed to identify and detect A. pleuropneumoniae, based on the 3＇-terminus of the species-specific apxlVA gene and the already existing species-specific primers in the omlA gene. Both 346-bp and 950-bp fragments could be simultaneously amplified from all A. pleuropneumoniae reference strains and isolates, and the species specificity of the assay was evaluated with a collection of ten strains representing eight different species bacteria including species normally found in the respiratory tracts of swine. All of these strains turned out negative in the multiplex PCR. All sequences of products of multiplex PCR randomly sampled were also correct. The sensitivity of the multiplex PCR was determined to be 10 pg of A. pleuropneumoniae DNA. The multiplex PCR and bacterial isolation were compared to determine their sensitivities by using experimentally infected pigs and clinical disease pigs. The multiplex PCR was more sensitive than bacterial isolation. The multiplex PCR was also evaluated on mixed bacterial cultures from clinical healthy pigs. 26/100 （26%） of the subclinically infected pigs were detected from clinical healthy pigs. The results indicate that the multiplex PCR assay is a sensitive, highly specific, and effective diagnostic tool for identification and detection of A. pleuropneumoniae.

关键词： multiplex PCR Actinobacillus pleuropneumoniae pig bacteria apxIVA and omlA genes

A regression approach for seismic first-break picking

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Petroleum Science 2024年第3期21卷 1584-1596页

作者： Huan Yuan san-Yi Yuan jie Wu Wen-Jing sang Yu-He Zhao CNPC Key Laboratory of Geophysical Exploration China University of Petroleum(Beijing)Beijing 102249China Research Institute of Petroleum Exploration&Development—Northwest(NWGI) Key Laboratory of reservoir characterization of CNPCPetroChinaLanzhou 730020GansuChina

The picking efficiency of seismic first breaks(FBs)has been greatly accelerated by deep learning(DL)***,the picking accuracy and efficiency of DL methods still face huge challenges in low signal-to-noise ratio(SNR)*** address this issue,we propose a regression approach to pick FBs based on bidirectional long short-term memory(Bi LSTM)neural network by learning the implicit Eikonal equation of 3D inhomogeneous media with rugged topography in the target *** employ a regressive model that represents the relationships among the elevation of shots,offset and the elevation of receivers with their seismic traveltime to predict the unknown FBs,from common-shot gathers with sparsely distributed *** from image segmentation methods which automatically extract image features and classify FBs from seismic data,the proposed method can learn the inner relationship between field geometry and *** addition,the predicted results by the regressive model are continuous values of FBs rather than the discrete ones of the binary *** picking results of synthetic data shows that the proposed method has low dependence on label data,and can obtain reliable and similar predicted results using two types of label data with large *** picking results of9380 shots for 3D seismic data generated by vibroseis indicate that the proposed method can still accurately predict FBs in low SNR *** subsequent stacked profiles further illustrate the reliability and effectiveness of the proposed *** results of model data and field seismic data demonstrate that the proposed regression method is a robust first-break picker with high potential for field application.

关键词： First-break picking Low signal-to-noiseratio Regression BiLSTM Traveltime Geometry Noisy seismic data

SG-NeRF:Sparse-Input Generalized Neural Radiance Fields for Novel View Synthesis

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Journal of Computer Science & Technology 2024年第4期39卷 785-797页

作者： Kuo Xu jie Li Zhen-Qiang Li Yang-jie cao School of Cyber Science and Engineering Zhengzhou UniversityZhengzhou 450002China Intelligent Big Data System(iBDSys)Lab Shanghai Jiao Tong UniversityShanghai 200240China School of Computer and Artificial Intelligence Zhengzhou UniversityZhengzhou 450001China

Traditional neural radiance fields for rendering novel views require intensive input images and pre-scene optimization,which limits their practical *** propose a generalization method to infer scenes from input images and perform high-quality rendering without pre-scene optimization named SG-NeRF(Sparse-Input Generalized Neural Radiance Fields).Firstly,we construct an improved multi-view stereo structure based on the convolutional attention and multi-level fusion mechanism to obtain the geometric features and appearance features of the scene from the sparse input images,and then these features are aggregated by multi-head attention as the input of the neural radiance *** strategy of utilizing neural radiance fields to decode scene features instead of mapping positions and orientations enables our method to perform cross-scene training as well as inference,thus enabling neural radiance fields to generalize for novel view synthesis on unseen *** tested the generalization ability on DTU dataset,and our PSNR(peak signal-to-noise ratio)improved by 3.14 compared with the baseline method under the same input *** addition,if the scene has dense input views available,the average PSNR can be improved by 1.04 through further refinement training in a short time,and a higher quality rendering effect can be obtained.

关键词： Neural Radiance Fields(NeRF) multi-view stereo(MVS) new view synthesis(NVS)

Impact pressure of waves generated by landslides on bank slopes

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Journal of Mountain Science 2024年第3期21卷 918-931页

作者： cao Ting WANG Pingyi QIU Zhenfeng LIU jie Key Laboratory of Hydraulic and Waterway Engineering of Ministry of Education Chongqing Jiaotong UniversityChongqing 400074China Engineering Research Center of Diagnosis Technology and Instruments of Hydro-Construction Chongqing Jiaotong UniversityChongqing 400074China

Impulse waves that are generated by landslides in narrow reservoir areas threaten the stability of buildings and bank *** discuss the action process and evolution law of the wave pressure on bank slopes,a three-dimensional physical model test that considers impulse waves generated by landslides was performed,and factors including landslide width,thickness,slope angles of the sliding surface,and bank slope angle were *** on wave forms on the bank slopes,wave pressure curve characteristics,and peak value,the action process of wave pressure could be divided into the following stages:maximum pulsating pressure stage,wave impact stage(when waves break),and stationary pulsation *** was found that wave breaking is dependent on the value of the surf similarity parameterξ.The distribution pattern of impact pressure decays linearly on both sides of the maximum impact pressure point,and the attenuation degree decreases when it attains 40%of the maximum ***,it is proposed that the prediction formula for the maximum effective impact pressure of the bank slope be related to the reciprocal of wave steepness,relative water depth,and slope *** prediction formula provides strong theoretical support for early safety warning and for predicting the bank slope under impulse waves generated by landslides.

关键词： Model test Impact pressure Action stage Breaking discrimination Distribution model

Efficient construction of calcium fluoride nanoaggregates for enhanced water vapor adsorption

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Particuology 2024年第6期89卷 191-197页

作者： Dan-Lei Yang Jing cao Rong-Kun Liu jie-Xin Wang School of Life Sciences and Chemical Engineering Jiangsu Second Normal UniversityNanjing210013China State Key Laboratory of Organic-Inorganic Composites Beijing University of Chemical TechnologyBeijing 100029China Research Center of the Ministry of Education for High Gravity Engineering and Technology Beijing University of Chemical TechnologyBeijing100029China Petrochemical Research Institute PetroChinaBeijing102206China

Calcium fluoride(CaF_(2))is an ideal adsorbent for the dehydration of gaseous hydrogen fluoride(HF)containing water *** this work,a novel CaF_(2)absorbent,spherical CaF_(2)nanoaggregates(NAs)with a closely packed structure,was proposed and efficiently fabricated by spray drying *** the building blocks of CaF_(2)NAs,the CaF_(2)nanoparticles(NPs)were prepared by the addition of excess calcium ions(Ca^(2+))or fluorine ions(F−)in the *** results indicated that the CaF_(2)NPs synthesized by excess Ca^(2+)and the corresponding NAs exhibited much better water vapor adsorption properties than their counterparts by excess F−,owing to higher zeta *** importantly,whether excess Ca^(2+)or F^(−),CaF_(2)NAs had further enhanced water vapor adsorption capacity compared to primary CaF_(2)NPs,possibly owing to their unique nano-micro secondary structures and higher surface *** work has great potential in the development of high-performance absorbents for separating moisture from corrosive gas HF.

关键词： CaF_(2)nanoaggregates Water vapor adsorption Spray drying

Influence of the penetration of adjacent X-section cast-in-place concrete(XCC)pile on the existing XCC pile in sand

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Journal of Zhejiang University-Science A(Applied Physics & Engineering) 2024年第7期25卷 557-572页

作者： Peng ZHOU Jianhui XU Changjie XU Guangwei cao jie CUP Xuanming DING School of Civil Engineering and Architecture State Key Laboratory of Performance Monitoring and Protecting of Rail Transit InfrastructureEast China Jiaotong UniversityNanchang330013China Guangdong Key Laboratory of Earthquake Engineering and Application Technology Earthquake Engineering Research&Test CenterGuangzhou UniversityGuangzhou510006China Key Laboratory of New Technology for the Construction of Cities in Mountain Areas College of Civil EngineeringChongqing UniversityChongqing400045China Zijin School of Geology and Mining Fuzhou UniversityFuzhou350116China

A series of small-scale 1g X-section cast-in-place concrete(XCC)pile-penetration model tests were conducted to study the effects of soil density and pile geometry on the lateral responses of an existing pile and the variations in surrounding soil *** results showed that the bending patterns of existing XCC piles varied with penetration *** lateral response of the existing pile was sensitive to the change in relative density and pile *** example,the bending moment of the existing pile increased along with these *** development of the radial stressσ′r/σ′v0 of the soil around an existing pile showed different trends at various depths during the penetration of the adjacent ***,the change in radial stress during the penetration of the XCC pile did not exhibit the“h/R effect”that was observed in the free-field soil,due to the shielding effect of the existing *** peak value of radial stressσ′r_max/σ′v0 decreased exponentially as the radial distance r/R *** attenuation ofσ′r_max/σ′v0 with r/R in the loose sand was faster than in the medium-dense or dense ***σ′r_max/σ′v0 at the same soil location increased with the cross-section geometry parameter.

关键词： X-section cast-in-place concrete(XCC)pile Test Penetration sand Lateral response Radial stress