Revealing the role of calcium ion intercalation of hydrated vanadium oxides for aqueous zinc-ion batteries

Exploring suitable high-capacity V_(2)O_(5)-based cathode materials is essential for the rapid advancement of aqueous zinc ion batteries(ZIBs).However,the typical problem of slow Zn^(2+)diffusion kinetics has severely limited the feasibility of such materials.In this work,unique hydrated vanadates(CaVO,BaVO)were obtained by intercalation of Ca^(2+)or Ba^(2+)into hydrated vanadium pentoxide.In the CaVO//Zn and BaVO//Zn batteries systems,the former delivered up to a 489.8 mAh g^(-1)discharge specific capacity at 0.1 A g^(-1).Moreover,the remarkable energy density of 370.07 Wh kg^(-1)and favorable cycling stability yard outperform BaVO,pure V_(2)O_(5),and many reported cathodes of similar ionic intercalation compounds.In addition,pseudocapacitance analysis,galvanostatic intermittent titration(GITT)tests,and Trasatti analysis revealed the high capacitance contribution and Zn^(2+)diffusion coefficient of CaVO,while an in-depth investigation based on EIS elucidated the reasons for the better electrochemical performance of CaVO.Notably,ex-situ XRD,XPS,and TEM tests further demonstrated the Zn^(2+)insertion/extraction and Zn-storage mechanism that occurred during the cycle in the CaVO//Zn battery system.This work provides new insights into the intercalation of similar divalent cations in vanadium oxides and offers new solutions for designing cathodes for high-capacity aqueous ZIBs.

Organoids revealed:morphological analysis of the profound next generation in-vitro model with artificial intelligence

In modern terminology,“organoids”refer to cells that grow in a specific three-dimensional(3D)environment in vitro,sharing similar structures with their source organs or tissues.Observing themorphology or growth characteristics of organoids through a microscope is a commonly used method of organoid analysis.However,it is difficult,time-consuming,and inaccurate to screen and analyze organoids only manually,a problem which cannot be easily solved with traditional technology.Artificial intelligence(AI)technology has proven to be effective in many biological and medical research fields,especially in the analysis of single-cell or hematoxylin/eosin stained tissue slices.When used to analyze organoids,AI should also provide more efficient,quantitative,accurate,and fast solutions.In this review,we will first briefly outline the application areas of organoids and then discuss the shortcomings of traditional organoid measurement and analysis methods.Secondly,we will summarize the development from machine learning to deep learning and the advantages of the latter,and then describe how to utilize a convolutional neural network to solve the challenges in organoid observation and analysis.Finally,we will discuss the limitations of current AI used in organoid research,as well as opportunities and future research directions.

Perilipin2 inhibits the replication of hepatitis B virus deoxyribonucleic acid by regulating autophagy under high-fat conditions

BACKGROUND Chronic hepatitis B virus(HBV)infection is often associated with increased lipid deposition in hepatocytes.However,when combined with non-alcoholic fatty liver disease or hyperlipidemia,it tends to have a lower HBV deoxyribonucleic acid(DNA)load.The relationship between lipid metabolism and HBV DNA replication and its underlying mechanisms are not well understood.AIM To investigate the relationship between lipid metabolism and HBV DNA replication and its underlying mechanisms.METHODS 1603 HBsAg-seropositive patients were included in the study.We first explored the relationship between patients'lipid levels,hepatic steatosis,and HBV DNA load.Also,we constructed an HBV infection combined with a hepatic steatosis cell model in vitro by fatty acid stimulation of HepG2.2.15 cells to validate the effect of lipid metabolism on HBV DNA replication in vitro.By knocking down and overexpressing Plin2,we observed whether Plin2 regulates autophagy and HBV replication.By inhibiting both Plin2 and cellular autophagy under high lipid stimulation,we examined whether the Plin2-autophagy pathway regulates HBV replication.RESULTS The results revealed that serum triglyceride levels,high-density lipoprotein levels,and hepatic steatosis ratio were significantly lower in the HBV-DNA high load group.Logistic regression analysis indicated that hepatic steatosis and serum triglyceride levels were negatively correlated with HBV-DNA load.Stratified analysis by HBeAg showed significant negative correlations between HBV-DNA load and hepatic steatosis ratio in both HBeAgpositive and HBeAg-negative groups.An in vitro cell model was developed by stimulating HepG2.2.15 cells with palmitic acid and oleic acid to study the relationship between HBV-DNA load and lipid metabolism.The results of the in vitro experiments suggested that fatty acid treatment increased lipid droplet deposition and decreased the expression of cell supernatant HBsAg,HBeAg,and HBV DNA load.Western blot and polymerase chain reaction analysis showed that fatty acid stimulation significantly induced Plin2 protein expression and inhibited the expression of hepatocyte autophagy proteins.Inhibition of Plin2 protein expression under fatty acid stimulation reversed the reduction in HBsAg and HBeAg expression and HBV DNA load induced by fatty acid stimulation and the inhibition of cellular autophagy.Knocking down Plin2 and blocking autophagy with 3-methyladenine(3-MA)inhibited HBV DNA replication.CONCLUSION In conclusion,lipid metabolism is a significant factor affecting HBV load in patients with HBV infection.The in vitro experiments established that fatty acid stimulation inhibits HBV replication via the Plin2-autophagy pathway.

Influencing factors for hepatic fat accumulation in patients with type 2 diabetes mellitus

BACKGROUND Non-alcoholic fatty liver disease has become the most common chronic liver disease worldwide,which originates from the accumulation of triglyceride(TG)in the liver.Patients with type 2 diabetes mellitus(T2DM)are considered to have a predisposition to hepatic steatosis.However,the influencing factors for hepatic fat accumulation in T2DM patients remain unclear.AIM To investigate the influencing factors for hepatic fat accumulation in T2DM patients.METHODS We enrolled 329 T2DM patients admitted to the Endocrinology Department of the First Affiliated Hospital of Soochow University,who underwent MR mDIXONQuant examination to quantify the hepatic fat fraction(HFF).According to body mass index(BMI),the patients were divided into normal weight,overweight,and obese groups.The differences in general statistics,biochemical parameters,islet function,and HFF were compared among the three groups.The associations between HFF and other parameters and the influences of various parameters on the severity of hepatic fat accumulation were analyzed.RESULTS The HFF of T2DM patients gradually increased in the normal weight,overweight,and obese groups(P<0.05).Spearman correlation analysis showed that in T2DM patients,HFF was negatively correlated with age and high-density lipoprotein cholesterol(P<0.05),whereas it was positively correlated with BMI,waist-hip ratio,fasting plasma glucose,alanine aminotransferase(ALT),aspartate aminotransferase,bilirubin,glutamyl transpeptidase,lactate dehydrogenase,albumin(ALB),uric acid(UA),total cholesterol,TG,low-density lipoprotein cholesterol(LDL-C),C-reactive protein,free triiodothyronine,fasting insulin,fasting C-peptide,and homeostasis model assessment of insulin resistance(P<0.05).Multiple linear regression analysis showed significant positive influences of BMI,ALT,LDL-C,UA,and ALB on HFF in T2DM patients(P<0.05).Binary logistic regression analysis showed that BMI,ALT,ALB,and LDL-C were independent risk factors for moderate to severe fatty liver in T2DM patients,and obesity increased the risk of being complicated with moderate to severe fatty liver by 4.03 times(P<0.05).CONCLUSION The HFF of T2DM patients increases with BMI.Higher BMI,ALT,ALB,and LDLC are independent risk factors for moderate to severe fatty liver in T2DM patients.

Influence of Sedimentation on Crystallization of Charged Colloidal Particles

The method of density matching between the solid and liquid phases is often adopted to effectively eliminate the effect of sedimentation of suspensions on dynamic behavior of a colloidal system. Experiments on crystallization of charged colloidal microspheres with di- ameter of 98 nm dispersed in density-matched and -unmatched media （mixtures of H20 and D20 in proper proportion） are compared to examine the influence of sedimentation. Reflection spectra of colloidal suspensions were used to monitor the crystallization process. Results showed that the crystal size of the density-unmatched （namely, in the presence of sedimentation） sample grew faster than that of the density-matched （in the absence of sedi- mentation） case at the initial stage of the crystallization, and then the latter overtook and outstripped the former. To explain these observations, we assume that in the settling of crystals sedimentation facilitates result in more particles getting into the crystal structures. However, as the crystals increase to varying sizes, the settling velocities become large and hydrodynamic friction strips off some particles from the delicate crystal structures. Overall, the sedimentation appears to accelerate the crystal size growth initially and then retard the growth. In addition, the crystal structures formed under microgravity were more closely packed than that in normal gravity.

Serum von Willebrand factor for early diagnosis of lung adenocarcinoma in patients with type 2 diabetes mellitus

BACKGROUND The elevation of plasma von Willebrand factor(vWF)has been proposed to be a predictor of lung cancer.Type 2 diabetes mellitus(T2DM)causes endothelial activation,resulting in the secretion of vWF.However,the role of vWF in patients with T2DM complicated with lung cancer remains unclear.AIM To investigate the clinical value of serum vWF as a tumor marker in patients with T2DM combined with lung adenocarcinoma in situ(AIS).METHODS This study enrolled 43 patients with T2DM combined with lung AIS(T2DM+AIS group),43 patients with T2DM alone(T2DM group),43 patients with lung AIS alone(AIS group),and 43 healthy volunteers(control group).The serum levels of vWF,insulin-like growth factor 1,and insulin-like growth factor binding protein 3 were determined.Multiple linear stepwise regression was performed to determine the correlations among variables.RESULTS Serum concentration of vWF in the T2DM+AIS group was significantly higher than those in the T2DM,AIS,and control groups(P<0.05).Serum vWF levels in the T2DM and AIS groups were significantly higher than that in the control group(P<0.05).There was no significant difference in serum vWF level between the T2DM and AIS groups.In the T2DM+AIS group,serum vWF was independently associated and positively correlated with serum levels of insulinlike growth factor 1 and insulin-like growth factor binding protein 3(P<0.05).CONCLUSION Serum vWF level may represent a novel biomarker for the early diagnosis of lung AIS.

Characterization of personal exposure concentration of fine particles for adults and children exposed to high ambient concentrations in Beijing, China

In China, the health risk from overexposure to particles is becoming an important public health concern. To investigate daily exposure characteristics to PM 2.5 with high ambient concentration in urban area, a personal exposure study was conducted for school children, and office workers in Beijing, China. For all participants （N = 114）, the mean personal 24-hr exposure concentration was 102.5, 14.7, 0.093, 0.528, 0.934, 0.174 and 0.703 μg/m 3 for PM 2.5 , black carbon, Mn, Al, Ca, Pb, and Fe. Children’s exposure concentrations of PM 2.5 were 4–5 times higher than those in related studies. The ambient concentration of PM 2.5 （128.5 μg/m 3 ） was significantly higher than the personal exposure concentration （P 0.05）, and exceed the reference concentration （25 μg/m 3 ） of WHO air quality guideline. Good correlation relationships and significant differences were identified between ambient concentration and personal exposure concentration. The relationships indicate that the ambient concentration is the main factor influencing personal exposure concentration, but is not a good indicator of personal exposure concentration. Outdoor activities （commute mode, exposure to heating, workday or weekend travel） influenced personal exposure concentrations significantly, but the magnitude of the influence from indoor activities （exposure to cooking） was masked by the high ambient concentrations.

钯在硝酸高铈铵溶液中的溶解工艺及宏观动力学研究

钯(Pd)是重要的战略金属,具有优异的理化性质,广泛应用于石油化工、汽车制造、航空航天和电子信息等领域。因其自然储量低,难以满足社会发展的需求,研究含钯二次资源回收技术,实现金属钯的循环利用具有重要意义。本工作针对传统湿法回收钯技术存在的酸耗高、NO_(x)排放量大等问题,提出了环境友好的硝酸高铈铵浸出新体系,考察了浸出剂浓度、酸度、添加剂浓度、反应温度、搅拌速度等因素对钯粉在硝酸高铈铵溶液中的溶解率的影响。结果表明,金属钯在混合体系中的溶解受到Pd表面吸附的显著影响。在没有Cl^(-)的情况下,金属钯表面吸附氧或者吸附NO_(3)^(-)形成了表面钝化,阻碍了Ce^(4+)与Pd原子的反应,5 h溶出率仅为0.2%。添加Cl^(-)后,通过竞争吸附能够有效破坏表面钝化层,构成突破点,从而加速钯的溶解反应。NO_(3)^(-)与Cl^(-)构成竞争吸附,更高浓度的硝酸根需要更高浓度的Cl^(-)启动溶解反应。优选浸出反应条件如下:硝酸高铈铵浓度1mol/L、硝酸浓度1 mol/L、盐酸浓度0.03 mol/L、反应温度80℃、搅拌速度200 r/min,在该条件下反应1.5 h钯粉的溶解率达到100%。钯粉在硝酸高铈铵溶液中的溶解过程符合化学反应控制的反应核缩减模型,表观活化能为58.7 kJ/mol,动力学方程为1-(1-x)^(1/3)=2264806e^(-58732/RT)t。

AtMCM10 promotes DNA replication-coupled nucleosome assembly in Arabidopsis

Minichromosome Maintenance protein 10(MCM10)is essential for DNA replication initiation and DNA elongation in yeasts and animals.Although the functions of MCM10 in DNA replication and repair have been well documented,the detailed mechanisms for MCM10 in these processes are not well known.Here,we identified AtMCM10 gene through a forward genetic screening for releasing a silenced marker gene.Although plant MCM10 possesses a similar crystal structure as animal MCM10,AtMCM10 is not essential for plant growth or development in Arabidopsis.AtMCM10 can directly bind to histone H3-H4 and promotes nucleosome assembly in vitro.The nucleosome density is decreased in Atmcm10,and most of the nucleosome density decreased regions in Atmcm10 are also regulated by newly synthesized histone chaperone Chromatin Assembly Factor-1(CAF-1).Loss of both AtMCM10 and CAF-1 is embryo lethal,indicating that AtM CM10 and CAF-1 are indispensable for replication-coupled nucleosome assembly.AtMCM10 interacts with both new and parental histones.Atmcm10 mutants have lower H3.1abundance and reduced H3K27me1/3 levels with releasing some silenced transposons.We propose that AtM CM10 deposits new and parental histones during nucleosome assembly,maintaining proper epigenetic modifications and genome stability during DNA replication.

植物RNA聚合酶V的染色质滞留机制

生命体遗传信息从DNA到RNA的传递需要由一类多亚基的RNA聚合酶来完成.在真核生物中,有3种保守的RNA聚合酶,即Pol Ⅰ (RNA Polymerase Ⅰ, RNA聚合酶I)、Pol Ⅱ和Pol Ⅲ,分别负责核糖体RNA、信使RNA以及转运RNA的合成[1].近年来,人们在植物中发现了另外两种植物特有的RNA聚合酶, Pol Ⅳ和Pol Ⅴ,丰富并扩展了人们对RNA聚合酶的认知[2,3].

纳米Co/rGO磁性复合吸附材料的制备及对Cu^2+的吸附性能

以改进Hummer法制备的薄片状氧化石墨烯(GO)为载体和模板负载钴离子,然后采用原位还原法制得纳米金属Co/石墨烯磁性复合吸附材料(Co/rGO),并将其应用于对Cu^2+的吸附和脱除,以期为高效可复用的铜离子脱除剂的合成与应用提供指导。实验结果证实,Co/rGO复合材料具有超顺磁性,能够很方便的使用磁铁进行分离并在无磁场情况下振荡分散。Co/rGO复合材料对Cu^2+具有稳定的吸附/脱附性能,实验条件下对Cu^2+的最大吸附容量达到117.5 mg/g且5 min内实现吸附平衡,远优于其原料GO的60 min吸附容量27.6 mg/g。本工作系统考察了NaOH加入量、络合剂种类、溶剂种类等关键因素对Co粒子在rGO载体上形貌和分布特性的影响,比较了不同合成条件下的复合材料对Cu^2+吸附效果的影响,并对优选条件下制备的Co/rGO复合材料进行了FT-IR,XRD,SEM表征。研究结果表明,纳米Co/rGO磁性材料对Cu^2+的吸附过程更符合Freundlich模型,属于多层吸附。室温下吸附焓ΔH=17.81 kJ/mol,吸附反应平衡常数K^θ=3.65。当初始Cu^2+浓度为39.22 mg/L时,对Cu^2+的吸附率为93.47%,五次吸附/脱附循环后吸附容量仍保持在初始值的94%,每次吸附后溶液中残余Cu^2+浓度均满足钴电解液对杂质铜离子的浓度去除要求(5 mg/L)或GB 8978-1996污水综合排放标准3级(2 mg/L),有望在相关领域发挥作用。

Expression of pathogenesis-related genes in cotton roots in response to Verticillium dahliae PAMP molecules

Verticillium wilt disease becomes a major threat to many economically important crops. It is unclear whether and how plant immunity takes place during cotton-Verticillium interaction due to the lack of marker genes. Taking advantage of cotton （Gossypium hirsutum） genome, we discovered pathogenesis-related （PR） gene families, which have been widely used as markers of immune responses in plants. To profile the expression of G. hirsutum PR genes in the process of plant immunity, we treated cotton roots with two immunogenic peptides, fig22 and nlp20 known as pathogen-associated molecular patterns, as well as three Verticillium dahliae-derived peptides, nlp20vd2, nlp23vd3, and nlp23vd4 which are highly identical to nlp20. Quantitative real-time PCR results revealed that 14 G. hirsutum PR gene （GhPR） families were induced or suppressed independently in response to fig22, nip20, nlp20va2, nlp23vd3, and nlp23vd4. Most GhPR genes are expressed highest at 3 h post incubation of immunogenic peptides. Compared to fig22 and nlp20, nlp20vd2 is more effective to trigger up-regulated expression of GhPR genes. Notably, both nlp23vd3 and nlp23vd4 are able to induce GhPR gene up-regulation, although they do not induce necrosis on cotton leaves. Thus, our results provide marker genes and new immunogenic peptides for further investigation of cotton-V, dahliae interaction.

有机单体空间构型对共轭微孔聚合物催化性能的影响

以TEMPO自由基功能化2,5-二溴-N-(2,2,6,6-四甲基哌啶)苯甲酰胺为有机单体,与平面三角构型(D3h对称性)的1,3,5-三乙炔苯和四面体构型(Td对称性)的四(4-乙炔基苯)甲烷分别通过Sonogashira-Hagihara偶联反应获得CMP-3-TEMPO和CMP-4-TEMPO。扫描电子显微镜照片(SEM)显示两种TEMPO功能化CMPs形貌上有显著差异,CMP-3-TEMPO为不规则的棒状和块状微米聚合物,CMP-4-TEMPO为尺寸较均一的微米球聚合物。固态电子顺磁共振谱(EPR)证明了CMP-3-TEMPO和CMP-4-TEMPO均含有TEMPO自由基。两种CMPs催化剂均可将5-羟甲基糠醛(5-HMF)选择性催化氧化成2,5-二甲酰基呋喃(2,5-DFF),但在催化效率和循环利用率方面存在显著差异。有机配体的空间立体构型对CMPs的形貌和催化性能有重要影响,选择具有空间立体构型的有机单体,所合成的CMP-4-TEMPO具有更稳定的结构,催化循环性能更优异。

Recognition of H3K9me1 by maize RNA-directed DNA methylation factor SHH2

RNA-directed DNA methylation(Rd DM) is a plant-specific de novo DNA methylation pathway,which has extensive cross-talk with histone modifications. Here, we report that the maize RdDM regulator SAWADEE HOMEODOMAIN HOMOLOG 2(SHH2) is an H3 K9 me1 reader. Our structural studies reveal that H3 K9 me1 recognition is achieved by recognition of the methyl group via a classic aromatic cage and hydrogen-bonding and salt-bridge interactions with the free protons of the mono-methyllysine. The di-and tri-methylation states disrupt the polar interactions, decreasing the binding affinity. Our study reveals a monomethyllysine recognition mechanism which potentially links RdDM to H3 K9 me1 in maize.

Conservation management decreases surface runoff and soil erosion

Conservation management practices e including agroforestry,cover cropping,no-till,reduced tillage,and residue return e have been applied for decades to control surface runoff and soil erosion,yet results have not been integrated and evaluated across cropping systems.In this study we collected data comparing agricultural production with and without conservation management strategies.We used a bootstrap resampling analysis to explore interactions between practice type,soil texture,surface runoff,and soil erosion.We then used a correlation analysis to relate changes in surface runoff and soil erosion to 13 other soil health and agronomic indicators,including soil organic carbon,soil aggregation,infil-tration,porosity,subsurface leaching,and cash crop yield.Across all conservation management practices,surface runoff and erosion had respective mean decreases of 67%and 80%compared with controls.Use of cover cropping provided the largest decreases in erosion and surface runoff,thus emphasizing the importance of maintaining continuous vegetative cover on soils.Coarse-and medium-textured soils had greater decreases in both erosion and runoff than fine-textured soils.Changes in surface runoff and soil erosion under conservation management were highly correlated with soil organic carbon,aggregation,porosity,infiltration,leaching,and yield,showing that conservation practices help drive important in-teractions between these different facets of soil health.This study offers the first large-scale comparison of how different conservation agriculture practices reduce surface runoff and soil erosion,and at the same time provides new insight into how these interactions influence the improvement or loss of soil health。

The H3K9me2-binding protein AGDP3 limits DNA methylation and transcriptional gene silencing in Arabidopsis

DNA methylation,a conserved epigenetic mark,is critical for tuning temporal and spatial gene expression.The Arabidopsis thaliana DNA glycosylase/lyase REPRESSOR OF SILENCING 1(ROS1)initiates active DNA demethylation and is required to prevent DNA hypermethylation at thousands of genomic loci.However,how ROS1 is recruited to specific loci is not well understood.Here,we report the discovery of Arabidopsis AGENET Domain Containing Protein 3(AGDP3)as a cellular factor that is required to prevent gene silencing and DNA hypermethylation.AGDP3 binds to H3K9me2 marks in its target DNA via its AGD12 cassette.Analysis of the crystal structure of the AGD12 cassette of AGDP3 in complex with an H3K9me2 peptide revealed that dimethylated H3 K9 and unmodified H3 K4 are specifically anchored into two different surface pockets.A histidine residue located in the methyllysine binding aromatic cage provides AGDP3 with pH-dependent H3K9me2 binding capacity.Our results uncover a molecular mechanism for the regulation of DNA demethylation by the gene silencing mark H3K9me2.

Interactions of Lump and Solitons to Generalized(2 + 1)-Dimensional Ito Systems

The(2 + 1)-dimensional Ito equation is extended to a general form including some nonintegrable effects via introducing generalized bilinear operators. It is pointed out that the nonintegrable(2 + 1)-dimensional Ito equation contains lump solutions and interaction solutions between lump and stripe solitons. The result shows that the lump soliton will be swallowed or arisen by a stripe soliton in a fixed time. Furthermore, by the interaction between a lump and a paired resonant stripe soliton, the lump will be transformed to an instanton or a rogue wave.

Effects of phosphatidylinositol 3-kinase inhibitor on humancervical carcinoma cells in vitro

Phosphatidylinositol 3-kinase(PI3K)is a crucial cell survival pathway implicated in tumorigenesis because of its role in stimulating cell proliferation and suppressing apoptosis.This study was to investigate the regulation of proliferation and apoptosis by LY294002,an inhibitor of PI3K in cervical cancer cells and the expression of FLICE-like inhibitory protein(c-FLIP)in vitro.Human cervical cancer HeLa cells were used in this experiment and cultured.The cultured cells were treated with LY294002 at different concentrations(10,25,50 and 100µmol/L)for 6,12,24,and 48 h before harvesting for evaluation.Cell viability was measured by 3-(4,5)-dimethylthiazol(-2-y1)-3,5-di-phenyltetrazoliumbromide(MTT)assay.Apoptosis was analyzed byflow cytometry.The expression of c-FLIP was detected by Western blot.Cell viability was inhibited by LY294002 significantly(P<0.05).Flow cytometry analysis revealed that cell apoptosis was significantly increased in the presence of LY294002 as compared with the control group.Although the expression of c-FLIP was increased in a short time,the expression of c-FLIP was markedly suppressed after the treatment of LY294002 for 48 h.These results suggested that the PI3K/Akt signal pathway might be involved in the regulation of cell apoptosis in cervical cancer cells.Moreover,the regulation of c-FLIP expression through PI3K/Akt signal pathway in cervical cancer cells was observed in vitro.

Comparison of gene expression responses of zebrafish larvae to Vibrio parahaemolyticus infection by static immersion and caudal vein microinjection

infection model for studying inflammation and the innate immune response.We investigated the genes and signaling pathways activated by static immersion and caudal vein microinjection infection using transcriptome profiling and reverse-transcription quantitative PCR(RT-qPCR)to compare the innate immune response in 3 days post-fertilization(dpf)zebrafish larvae infected by Vibrio parahaemolyticus Vp13 strain.The median lethal dose(LD50)values at 96 h following immersion and microinjection were 3.63×107 CFU/mL and 5.76×102 CFU/nL,respectively.An innate immune response was initiated after 2 h of incubation with the respective LD50 for each infection method.Six hundred and two genes in the immersion group and 359 genes in the microinjection group were activated and differentially expressed post-infection.Sixty-three Gene Ontology(GO)terms and four Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways were significantly enriched in the immersion group,compared with only three GO terms and no KEGG pathways in the microinjection group.Two genes,tnfb and ccl20a.3,were significantly up-regulated in both groups.We speculated that immersion infection may affect initial dorsal determination,cytochromes,and fatty acid-binding proteins,as well as inflammation,while microinjection infection may mainly directly affect the immune response.Infection with doses>LD50(1.09×109 CFU/mL and 1.09×103 CFU/nL by immersion and microinjection,respectively)caused more significant up-regulation of il11a,tnfa,tnfb,il1b,ccl34a.4,ccl20a.3,irak3,cxcl18b,and ccl35.1,suggesting that in addition to the classical innate immunity genes tnfa,tnfb,il1b,and il6,the genes il11a,ccl34a.4,ccl20a.3,cxcl18b,and ccl35.1 were also important for defending against Vp13 infection.These findings highlight the genes involved in the responses of zebrafish to Vp13 infection via different routes and doses,and thus provide the basis for further analyses of immune response signaling pathways.