The differential impact of face distractors on visual working memory across encoding and delay stages.

External distractions often occur when information must be retained in visual working memory (VWM)-a crucial element in cognitive processing and everyday activities. However, the distraction effects can differ if they occur during the encoding rather than the delay stages. Previous research on these effects used simple stimuli (e.g., color and orientation) rather than considering distractions caused by real-world stimuli on VWM. In the present study, participants performed a facial VWM task under different distraction conditions across the encoding and delay stages to elucidate the mechanisms of distraction resistance in the context of complex real-world stimuli. VWM performance was significantly impaired by delay-stage but not encoding-stage distractors (Experiment 1). In addition, the delay distraction effect arose primarily due to the absence of distractor process at the encoding stage rather than the presence of a distractor during the delay stage (Experiment 2). Finally, the impairment in the delay-distraction condition was not due to the abrupt appearance of distractors (Experiment 3). Taken together, these findings indicate that the processing mechanisms previously established for resisting distractions in VWM using simple stimuli can be extended to more complex real-world stimuli, such as faces.

ATG5 nonautophagically regulates inflammation and differentiation in mouse embryonic stem cells.

Embryonic stem cells (ESCs), with abilities of infinite proliferation (self-renewal) and to differentiate into distinct cell types (pluripotency), show attenuated inflammatory response against cytokines or pathogens, which is recognized as a unique characteristic of ESCs compared with somatic cells. However, the underlying molecular mechanisms remain unclear, and whether the attenuated inflammatory state is involved in ESC differentiation is completely unknown. Our recent study demonstrated that macroautophagy/autophagy-related protein ATG5 inhibits the inflammatory response of mouse ESCs (MmESCs) by promoting the degradation of BTRC/ß-TrCP1 and further the downregulation of NFKB/NF-κB signaling. In addition, maintenance of an attenuated inflammation status in MmESCs is required for their differentiation. In conclusion, ATG5 is a key regulator for the regulation of inflammatory response and differentiation of MmESCs.

ATG5 attenuates inflammatory signaling in mouse embryonic stem cells to control differentiation.

Attenuated inflammatory response is a property of embryonic stem cells (ESCs). However, the underlying mechanisms are unclear. Moreover, whether the attenuated inflammatory status is involved in ESC differentiation is also unknown. Here, we found that autophagy-related protein ATG5 is essential for both attenuated inflammatory response and differentiation of mouse ESCs and that attenuation of inflammatory signaling is required for mouse ESC differentiation. Mechanistically, ATG5 recruits FBXW7 to promote ubiquitination and proteasome-mediated degradation of ß-TrCP1, resulting in the inhibition of nuclear factor κB (NF-κB) signaling and inflammatory response. Moreover, differentiation defects observed in ATG5-depleted mouse ESCs are due to ß-TrCP1 accumulation and hyperactivation of NF-κB signaling, as loss of ß-TrCP1 and inhibition of NF-κB signaling rescued the differentiation defects. Therefore, this study reveals a previously uncharacterized mechanism maintaining the attenuated inflammatory response in mouse ESCs and further expands the understanding of the biological roles of ATG5.

A universal design of restructured dimer antigens: Development of a superior vaccine against the paramyxovirus in transgenic rice.

The development of vaccines, which induce effective immune responses while ensuring safety and affordability, remains a substantial challenge. In this study, we proposed a vaccine model of a restructured "head-to-tail" dimer to efficiently stimulate B cell response. We also demonstrate the feasibility of using this model to develop a paramyxovirus vaccine through a low-cost rice endosperm expression system. Crystal structure and small-angle X-ray scattering data showed that the restructured hemagglutinin-neuraminidase (HN) formed tetramers with fully exposed quadruple receptor binding domains and neutralizing epitopes. In comparison with the original HN antigen and three traditional commercial whole virus vaccines, the restructured HN facilitated critical epitope exposure and initiated a faster and more potent immune response. Two-dose immunization with 0.5 µg of the restructured antigen (equivalent to one-127th of a rice grain) and one-dose with 5 µg completely protected chickens against a lethal challenge of the virus. These results demonstrate that the restructured HN from transgenic rice seeds is safe, effective, low-dose useful, and inexpensive. We provide a plant platform and a simple restructured model for highly effective vaccine development.

Polymersomes in Drug Delivery─From Experiment to Computational Modeling.

Polymersomes, composed of amphiphilic block copolymers, are self-assembled vesicles that have gained attention as potential drug delivery systems due to their good biocompatibility, stability, and versatility. Various experimental techniques have been employed to characterize the self-assembly behaviors and properties of polymersomes. However, they have limitations in revealing molecular details and underlying mechanisms. Computational modeling techniques have emerged as powerful tools to complement experimental studies and enabled researchers to examine drug delivery mechanisms at molecular resolution. This review aims to provide a comprehensive overview of the state of the art in the field of polymersome-based drug delivery systems, with an emphasis on insights gained from both experimental and computational studies. Specifically, we focus on polymersome morphologies, self-assembly kinetics, fusion and fission, behaviors in flow, as well as drug encapsulation and release mechanisms. Furthermore, we also identify existing challenges and limitations in this rapidly evolving field and suggest possible directions for future research.

Rice-produced classical swine fever virus glycoprotein E2 with herringbone-dimer design to enhance immune responses.

Pestiviruses, including classical swine fever virus, remain a concern for global animal health and are responsible for major economic losses of livestock worldwide. Despite high levels of vaccination, currently available commercial vaccines are limited by safety concerns, moderate efficacy, and required high doses. The development of new vaccines is therefore essential. Vaccine efforts should focus on optimizing antigen presentation to enhance immune responses. Here, we describe a simple herringbone-dimer strategy for efficient vaccine design, using the classical swine fever virus E2 expressed in a rice endosperm as an example. The expression of rE2 protein was identified, with the rE2 antigen accumulating to 480 mg/kg. Immunological assays in mice, rabbits, and pigs showed high antigenicity of rE2. Two immunizations with 284 ng of the rE2 vaccine or one shot with 5.12 µg provided effective protection in pigs without interference from pre-existing antibodies. Crystal structure and small-angle X-ray scattering results confirmed the stable herringbone dimeric conformation, which had two fully exposed duplex receptor binding domains. Our results demonstrated that rice endosperm is a promising platform for precise vaccine design, and this strategy can be universally applied to other Flaviviridae virus vaccines.

Safety assessment of organic micropollutants in reclaimed water: Chemical analyses, ecological risk assessments, and in vivo endocrine-disrupting studies.

Reclaimed water from municipal wastewater has great potential in mitigating the water resource crisis, while the inevitable residue of organic micropollutants (OMPs) challenges the safety of reclaimed water reuse. Limited information was available regarding the overall adverse effects of mixed OMPs in reclaimed water, especially the endocrine-disrupting effects on living organisms. Herein, chemical monitoring in two municipal wastewater treatment plants showed that 31 of 32 candidate OMPs including polycyclic aromatic hydrocarbons (PAHs), phenols, pharmaceuticals and personal care products (PPCPs) were detected in reclaimed water, with a concentration ranging from ng/L to µg/L. Then, based on the risk quotient value, phenol, bisphenol A, tetracycline, and carbamazepine were ranked as high ecological risks. Most PAHs and PPCPs were quantified as medium and low risks, respectively. More importantly, using aquatic vertebrate zebrafish as an in vivo model, the endocrine-disrupting potentials of OMP mixtures were comprehensively characterized. We found that a realistic exposure to reclaimed water induced estrogen-like endocrine disruption and hyperthyroidism in zebrafish, abnormal expression of genes along the hypothalamus-pituitary-thyroid (-gonad) axes, reproductive impairment, and transgenerational toxicity. Based on the chemical analyses, risk quotient calculations, and biotoxicity characterization, this study contributed to understanding the ecological risks of reclaimed water and developing the control standards for OMPs. In addition, application of the zebrafish model in this study also highlighted the significance of in vivo biotoxicity test in water quality evaluation.

The effect of sad mood on early sensory event-related potentials to task-irrelevant faces.

It has been shown that the perceiver's mood affects the perception of emotional faces, but it is not known how mood affects preattentive brain responses to emotional facial expressions. To examine the question, we experimentally induced sad and neutral mood in healthy adults before presenting them with task-irrelevant pictures of faces while an electroencephalography was recorded. Sad, happy, and neutral faces were presented to the participants in an ignore oddball condition. Differential responses (emotional - neutral) for the P1, N170, and P2 amplitudes were extracted and compared between neutral and sad mood conditions. Emotional facial expressions modulated all the components, and an interaction effect of expression by mood was found for P1: an emotional modulation to happy faces, which was found in neutral mood condition, disappeared in sad mood condition. For N170 and P2, we found larger response amplitudes for both emotional faces, regardless of the mood. The results add to the previous behavioral findings showing that mood already affects low-level cortical feature encoding of task-irrelevant faces.

Pain modulates early sensory brain responses to task-irrelevant emotional faces.

BACKGROUND: Pain can have a significant impact on an individual's life, as it has both cognitive and affective consequences. However, our understanding of how pain affects social cognition is limited. Previous studies have shown that pain, as an alarm stimulus, can disrupt cognitive processing when focal attention is required, but whether pain also affects task-irrelevant perceptual processing remains unclear. METHODS: We examined the effect of laboratory-induced pain on event-related potentials (ERPs) to neutral, sad and happy faces before, during and after a cold pressor pain. ERPs reflecting different stages of visual processing (P1, N170 and P2) were analysed. RESULTS: Pain decreased the P1 amplitude for happy faces and increased the N170 amplitude for happy and sad faces compared to the pre-pain phase. The effect of pain on N170 was also observed in the post-pain phase. The P2 component was not affected by pain. CONCLUSIONS: Our results suggest that pain alters both featural (P1) and structural face-sensitive (N170) visual encoding of emotional faces, even when the faces are irrelevant to the task. While the effect of pain on initial feature encoding seemed to be disruptive and specific to happy faces, later processing stages showed long-lasting and increased activity for both sad and happy emotional faces. SIGNIFICANCE: The observed alterations in face perception due to pain may have consequences for real-life interactions, as fast and automatic encoding of facial emotions is important for social interactions.

Alterations in working memory maintenance of fearful face distractors in depressed participants: An ERP study.

Task-irrelevant threatening faces (e.g., fearful) are difficult to filter from visual working memory (VWM), but the difficulty in filtering non-threatening negative faces (e.g., sad) is not known. Depressive symptoms could also potentially affect the ability to filter different emotional faces. We tested the filtering of task-irrelevant sad and fearful faces by depressed and control participants performing a color-change detection task. The VWM storage of distractors was indicated by contralateral delay activity, a specific event-related potential index for the number of objects stored in VWM during the maintenance phase. The control group did not store sad face distractors, but they automatically stored fearful face distractors, suggesting that threatening faces are specifically difficult to filter from VWM in non-depressed individuals. By contrast, depressed participants showed no additional consumption of VWM resources for either the distractor condition or the non-distractor condition, possibly suggesting that neither fearful nor sad face distractors were maintained in VWM. Our control group results confirm previous findings of a threat-related filtering difficulty in the normal population while also suggesting that task-irrelevant non-threatening negative faces do not automatically load into VWM. The novel finding of the lack of negative distractors within VWM storage in participants with depressive symptoms may reflect a decreased overall responsiveness to negative facial stimuli. Future studies should investigate the mechanisms underlying distractor filtering in depressed populations.

Preparation and evaluation of chitosan- and hyaluronic acid-grafted pullulan succinate films for skin wound healing.

A novel wound dressing that possesses antibacterial properties and accelerates skin wound repair was developed by physically blending hyaluronic acid-grafted pullulan succinate (HA-st-Pu) with chitosan (CS). The HA-st-Pu polymer was synthesized and characterized, and then CS/HA-st-Pu film dressings were prepared by a freeze-drying method. The novel film wound dressings exhibited a three-dimensional cavity structure under scanning electron microscopy (SEM) and a better swelling ratio than CS, HA and Pu alone, absorbing a large amount of liquid and effectively maintaining the moist environment of the wound. CS/HA-st-Pu materials had no cytotoxicity and increased cell proliferation when coincubated with L929 cells. Moreover, CS/HA-st-Pu wound dressings exhibited a certain antibacterial capability against E. coli and S. aureus. In rat skin wound healing, CS/HA-st-Pu film dressings outperformed both the control and market band-aid groups with respect to the reduction of inflammation and acceleration of wound closure.

Characterization of Neutralizing Monoclonal Antibodies and Identification of a Novel Conserved C-Terminal Linear Epitope on the Hemagglutinin Protein of the H9N2 Avian Influenza Virus.

The H9N2 avian influenza virus (AIV) remains a serious threat to the global poultry industry and public health. The hemagglutinin (HA) protein is an essential protective antigen of AIVs and a major target of neutralizing antibodies and vaccines. Therefore, in this study, we used rice-derived HA protein as an immunogen to generate monoclonal antibodies (mAbs) and screened them using an immunoperoxidase monolayer assay and indirect enzyme-linked immunosorbent assay. Eight mAbs reacted well with the recombinant H9N2 AIV and HA protein, four of which exhibited potent inhibitory activity against hemagglutination, while three showed remarkable neutralization capacities. Western blotting confirmed that two mAbs bound to the HA protein. Linear epitopes were identified using the mAbs; a novel linear epitope, 480HKCDDQCM487, was identified. Structural analysis revealed that the novel linear epitope is located at the C-terminus of HA2 near the disulfide bond-linked HA1 and HA2. Alignment of the amino acid sequences showed that the epitope was highly conserved among multiple H9N2 AIV strains. The results of this study provide novel insights for refining vaccine and diagnostic strategies and expand our understanding of the immune response against AIV.

Early- and whole-life exposures to florfenicol disrupts lipid metabolism and induces obesogenic effects in zebrafish (Danio rerio).

Florfenicol (FF), a widely used veterinary antibiotic, has been frequently detected in both aquatic environments and human body fluids. As a result, there is a growing concern on its health risks. Previous studies have revealed various toxicities of FF on animals, while there are relatively limited researches on its metabolic toxicity. Herein, by employing zebrafish as an in vivo model, endpoints at multiple levels of biological organization were measured to investigate the metabolic toxicity, especially disturbances on lipid metabolism, of this emerging pollutant. Our results indicated that early-life exposure (from 2 h past fertilization (hpf) to 15 days past fertilization (dpf)) to FF significantly increased body mass index (BMI) values, staining areas of visceral lipids, and triacylglycerol (TAG) and total cholesterol (TC) contents of larvae. Further, by analyzing expression patterns of genes encoding key proteins regulating lipid metabolism, our data suggested that promoted intestinal absorption and hepatic de novo synthesis of lipids, suppressed TAG decomposition, and inhibited FFA oxidation all contributed to TAG accumulation in larvae. Following whole-life exposure (from 2 hpf to 120 dpf), BMI values, TAG and TC contents all increased significantly in males, and significant increases of hepatic TAG levels were also observed in females. Moreover, FF exposure interfered with lipid homeostasis of males and females in a gender-specific pattern. Our study revealed the obesogenic effects of FF at environmentally relevant concentrations (1, 10, and 100 µg/L) and therefore will benefit assessment of its health risks. Additionally, our results showed that FF exposure caused a more pronounced obesogenic effect in zebrafish larvae than adults, as suggested by significant increases of all endpoints at individual, tissular, and molecular levels in larvae. Therefore, our study also advances the application of zebrafish larval model in assessing metabolic toxicity of chemicals, due to the higher susceptibility of larvae than adults.

Design and evaluation of glimepiride hydrogel for transdermal delivery.

The solubility of glimepiride (GM) was improved from 1.6 µg/mL to 22.0 mg/mL when GM and meglumine (MU) complexes were prepared. Therefore, transdermal hydrogels of GM Carbopol (GM-CP) and GM hydroxypropyl methylcellulose pullulan (GM-HPMC-Pu) were prepared successfully utilizing the improved drug solubility by GM-MU. Based on a single factor experiment and response surface methodology, two kinds of hydrogel formulations were optimized by drug release studies in vitro. The optimized GM-CP hydrogel was composed of GM, a mixture of azone and oleic acid (1:1, 2.6%, v/v) and carbopol 940 (1%, w/v). The GM-HPMC-Pu hydrogel was developed using GM, HPMC (3.5%, w/v), Pu (1.5%, w/v), glycerol (5%, v/v), azone (2.9%, v/v) and oleic acid (2.6%, v/v). The study of hydrogels in vivo was performed using rabbits. The results indicated that the drug could sustain release from GM-CP or GM-HPMC-Pu hydrogel and maintain the high plasma concentration for 48 h. Compared with commercial GM tablets, the relative bioavailability of GM-CP and GM-HPMC-Pu hydrogel reached 48% and 133%, respectively. Moreover, the drug release in vitro could well predict its absorption in vivo. There was a good correlation (R2≥0.966) in GM hydrogel between the drug release in vitro and transdermal absorption in vivo. Therefore, a novel GM hydrogel dosage form may be considered to design.

An Improved Immunochromatographic Strip Based on Plant-Derived E2 for Detection of Antibodies against Classical Swine Fever Virus.

Vaccination is an effective method to control the spread of classical swine fever virus (CSFV), which is a major cause of economic losses to the swine industry. Although serological detection assays are commonly used to assess immune status, current methods for monitoring of antibodies (Abs) are time-consuming, expensive, and require cell culture and virus manipulation. To address these problems, the E2 protein of CSFV was expressed in transgenic rice seeds as a labeled antigen for the development of an immunochromatographic test strip (ICTS) for rapid, precise, and cost-effective detection of Abs. The ICTS has a reasonable sensitivity of 1:128,000 for detection of serum Abs against CSFV and no cross-reactivity with Abs of other porcine viruses. The similarity of the results between the proposed ICTS and a commercial enzyme-linked immunosorbent assay was 94.1% (128/136) for detection of serum Abs from immunized animals and 92.3% (72/78) for detection of maternally derived Abs. The proposed assay was successfully used to monitor Abs against E2 of both pigs and rabbits immunized with a live attenuated vaccine or an E2 subunit vaccine. The results confirmed that the ICTS can be applied to detect Ab levels in animals with different immunological backgrounds. The ICTS based on plant-derived E2 is a relatively inexpensive, rapid, and accurate assay for detection of Abs against CSFV and avoids the risk of contamination by animal products. IMPORTANCE The E2 protein of classical swine fever virus (CSFV) was expressed in transgenic rice endosperms as a diagnostic antigen for use with a rapid colloidal gold assay for the detection of antibodies (Abs) against CSFV. This improved test was used to monitor Abs against the E2 protein in both pigs and rabbits immunized with a live attenuated vaccine or E2 subunit vaccine. The assay successfully detected Ab levels in serum samples from piglets with different immunological backgrounds. In contrast to current E2 protein-based diagnostic methods using Escherichia coli or insect cells as expression systems, plant-derived E2 avoids the limitations of low immunogenicity of eukaryotic expression systems and potential contamination of fetal bovine serum with bovine viral diarrhea virus in cell culture.

Risk and molecular mechanisms for boscalid resistance in Penicillium digitatum.

The succinate dehydrogenase inhibitor (SDHI) fungicide boscalid is an excellent broad-spectrum fungicide but has not been registered in China to control Penicillium digitatum, the causal agent of green mold of citrus. The present study evaluated the risk and molecular mechanisms for boscalid resistance in P. digitatum. Resistance induction with four arbitrarily selected sensitive isolates of P. digitatum by ultraviolet (UV) irradiation on conidia plated on boscalid-amended potato dextrose agar (PDA) and consecutive growing on boscalid-amended PDA produced five highly resistant isolates with EC50 values greater than 1000 µg/mL and two resistant isolates with EC50 lower than 200 µg/mL. Boscalid resistance of the five mutants with EC50 values above 1000 µg/mL was stable after successive transfers on PDA for 16 generations. However, for the other two mutants with EC50 lower than 200 µg/mL, the EC50 values decreased significantly after successive transfers. There was significant cross-resistance between boscalid and carboxin (r = 0.925, P < 0.001), but no significant cross-resistance was detected between boscalid and fludioxonil (r = 0.533，P = 0.095) or between boscalid and prochloraz (r = -0.543，P = 0.088). The seven resistant mutants varied greatly in the mycelia growth, sporulation, pathogenicity, and sensitivities to exogenous stresses including NaCl, salicylhydroxamic acid (SHAM), and H2O2. Alignment of the deduced amino acid sequence showed that there was no point mutation in the target enzyme succinate dehydrogenase (Sdh) subunits SdhA, SdhC, or SdhD in each of the seven resistant mutants, and the mutation of a conserved histidine residue to tyrosine (H243Y) in the subunit SdhB (i.e., iron­sulfur protein) occurred in only three highly resistant isolates. Molecular docking indicated that mutation H243Y could not prevent the binding of boscalid into the quinone-binding site of SDH in the presence of the heme moiety. However, for SDH without the heme moiety, boscalid could bind into a deeper site with a much higher affinity, and the mutation H243Y spatially blocked the docking of boscalid into the deeper site. This may be the molecular mechanism for boscalid resistance caused by SdhB-H243Y mutation.

[Expression of NDV HN protein in rice and development of a semi-quantitative rapid method for detection of antibodies].

The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log2, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.

miR-148-3p inhibits gastric cancer cell malignant phenotypes and chemotherapy resistance by targeting Bcl2.

Gastric cancer(GC) is the fourth most common cancer in the world. This work was designed to explore the biological effects of miR-148-3p on GC. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was utilized to analyze the mRNA expression of miR-148-3p in GC cell lines. The mimics and inhibitors of miR-148-3p, was carefully transfected into GC cells to up-regulate or down-regulate miR-148-3p expression. Observe the effect on miR-148-3p expression change to GC cell proliferation, colony formation, tumorigenesis, chemotherapy sensitivity, trans-well migration and invasion. Use online database tool to predict the miR-148-3p promising targets, and be verified via RT-qPCR, Western blot and luciferase report. We found that miR-148-3p expression level in GC cells was markedly down-regulated (P <0.05), as compared with human normal gastric mucosal cells GES-1. Otherwise, miR-148-3p overexpression could effectively inhibit the cell proliferation, cell cycle progress, colony formation, anti-apoptosis, anti-migration and anti-invasion in gastric cancer cells, whereas miR-148-3p inhibition exhibited the opposite phenomenon (P<0.05). Further research revealed that Bcl2 set as a direct downstream target of miR-148-3p. Our study firstly confirmed that, miR-148-3p might play a crucial role in tumorigenesis, as well as development of gastric cancer by targeting Bcl2, and could become a promising target for gastric cancer treatment.

A novel linear epitope at the C-terminal region of the classical swine fever virus E2 protein elicits neutralizing activity.

Classical swine fever virus (CSFV) is a member of the genus Pestivirus, which causes serious economic losses. The re-emergence of the disease in Japan in 2018 has increased awareness of CSFV. In this study, Balb/c mice were immunized with plant-derived E2 protein, and four monoclonal antibodies (mAbs) 4B11, 7B3, 11A5 and 6F3 were generated. Two of these mAbs, 4B11 and 7B3, effectively blocked CSFV infection of PK-15 cells. Both mAbs recognized a novel linear epitope, 256CLIGNTTVKVHASDER271. The neutralizing ability of anti-CSFV serum decreased 63%, when pre-incubated with the linear peptide at 200 µg/mL. Structural analysis showed that this linear epitope is present at the border of Domain C and Domain D on the surface of the E2 protein. Alignment of amino acid sequences showed that the epitope was conserved in different subgroups of CSFV but not in other members of the Pestivirus genus. Consistently with the analysis above, this epitope distinguished antibodies against CSFV from those against bovine viral diarrhea virus (BVDV). Our study provides an ideal candidate peptide for new vaccine design and differential diagnosis of CSFV. These findings will contribute to the control and eradication of classical swine fever.