Development of Hypoallergenic Derivatives of Cra a 1 with B Cell Epitope Deletion and T Cell Epitope Retention.

Tropomyosin was reported as an important allergen in Crassostrea angulata and designated as Cra a 1. The localization of the T cell epitopes and the reduction of the immunoreactivity of Cra a 1 are still lacking. In this study, four T cell epitopes were identified by using wild-type Cra a 1 (wtCra a 1)-immunized mouse splenocytes cultured with synthetic peptides. The immunoreactivity was maintained after chemical denaturation treatment, indicating that the linear epitope is an immunodominant epitope of wtCra a 1. Furthermore, the hypoallergenic derivative (mCra a 1) was developed by the deletion of linear B cell epitopes and retention of T cell epitopes. mCra a 1 could stimulate CD4+T cell proliferation and upregulate interleukin-10 secretion. Overall, basophil activation by mCra a 1 was low, but its ability to induce T cell proliferation was retained, suggesting that mCra a 1 may serve as a viable candidate for treating oyster allergy.

Potential mechanisms of cancer stem-like progenitor T-cell bio-behaviours.

In situations involving continuous exposure to antigens, such as chronic infections or cancer, antigen-specific CD8+ T cells can become dysfunctional or exhausted. This change is marked by increased expression levels of inhibitory receptors (PD-1 and Tim-3). Stem-like progenitor exhausted (Tpex) cells, a subset of exhausted cells that express TCF-1 and are mainly found in the lymph nodes, demonstrate the ability to self-renew and exhibit a high rate of proliferation. Tpex cells can further differentiate into transitional intermediate exhausted (Tex-int) cells and terminally exhausted (Tex-term) cells. Alternatively, they can directly differentiate into Tex-term cells. Tpex cells are the predominant subset that respond to immune checkpoint inhibitors (ICI), making them a prime candidate for improving the efficacy of ICI therapy. This review article aimed to present the latest developments in the field of Tpex formation, expansion, and differentiation in the context of cancer, as well as their responses to ICIs in cancer immunotherapy. Consequently, it may be possible to develop novel treatments that exclusively target Tpex cells, thus improving overall treatment outcomes. KEY POINTS: Tpex cells are located in lymph nodes and TLS. Several pathways control the differentiation trajectories of Tpex cells, including epigenetic factors, transcription factors, cytokines, age, sex, etc.

NR4A1 transcriptionally regulates the differentiation of stem-like CD8+ T cells in the tumor microenvironment.

CD8+ T cells are rendered exhausted in tumor and chronic infection. Among heterogeneous exhausted T cells, a subpopulation of progenitor-like (Tpex) cells have been found important for long-term tumor or pathogen control and are also the main responders in immunotherapy. Using an RFP reporter mouse for the orphan nuclear receptor NR4A1, originally characterized as critical in T cell dysfunction, we discover that the reporter is highly expressed in Tpex cells in tumor and chronic infection. Enforced expression of Nr4a1 promotes Tpex cell accumulation, whereas tumor control is improved after Nr4a1 deletion, associated with increased effector function but decreased long-term maintenance of CD8+ T cells. Integrating chromatin immunoprecipitation sequencing (ChIP-seq) and RNA sequencing (RNA-seq) analysis, NR4A1 is found to bind and promote the expression of Tpex-related genes, as well as suppress terminal differentiation-associated genes. This study therefore has identified a key role of NR4A1 in Tpex regulation and provides a promising target for immunotherapy.

Visible-light-induced N-alkylation of anilines with 4-hydroxybutan-2-one.

The synthesis of amines through N-alkylation is particularly attractive. Herein, a strategy for visible-light-induced N-alkylation of anilines with 4-hydroxybutan-2-one was developed in the presence of NH4Br, which avoid the use of metals, bases and ligands. In addition, gram-scale experiments proved that the system has the potential to be scaled.

Metabolomics analysis of physicochemical properties associated with freshness degradation in frozen Antarctic krill (Euphausia superba).

This study aimed to determine the effect of different frozen temperatures during storage on the quality of Antarctic krill (Euphausia superba) and assess the change at the metabolite level via a combination of physicochemical property analysis, liquid chromatography-tandem mass spectrometry (LC-MS) based non-targeted metabolomics profiling. Regarding samples stored at -20 °C, the expressions of 7055 metabolites were elevated, while 2313 were downregulated. Lipids and lipid molecules had the highest proportion of differential metabolites. A total of 432 discriminatory metabolites with Kyoto Encyclopedia of Genes and Genomes (KEGG) IDs was obtained. We also observed that the concentrations of differential bitter free amino acids (FAAs) and oxidation products of arachidonic and linoleic acid increased. Moreover, as the storage temperature increased, the freshness, umami, and sweetness components were considerably reduced. Furthermore, results indicated that the color, pH and water-holding capacity (WHC) were potential indicators of quality deterioration, while inosinic acid was a probable biomarker for umami degradation of frozen Antarctic krill. In conclusion, this study demonstrates that storage at lower temperatures can be beneficial for maintaining the freshness of Antarctic krill from macro and micro perspectives.

Mutation of Disulfide Bond Sites Reduces the Immunoreactivity of Cra a 4 by Changing the Structural Characteristics.

Sarcoplasmic calcium-binding protein (Cra a 4) from Crassostrea angulata belongs to the EF-hand superfamily, and understanding of its structure-allergenicity relationship is still insufficient. In this study, chemical denaturants were used to destroy the structure of Cra a 4, showing that disruption of the structure reduced its IgG-/IgE-binding activity. To explore which critical amino acid site affects the allergenicity of Cra a 4, the mutants were obtained by site-directed mutations in the disulfide bonds site (C97), conformational epitopes (I105, D114), or Ca2+-binding region (D106, D110) and their IgG-/IgE-binding activity was reduced significantly using serological tests. Notably, C97A had the lowest immunoreactivity. In addition, two conformational epitopes of Cra 4 were verified. Meanwhile, the increase of the α-helical content, surface hydrophobicity, and surface electrostatic potential of C97A affected its allergenicity. Overall, the understanding of the structure-allergenicity relationship of Cra a 4 allowed the development of a hypoallergenic mutant.

Notch Signaling Inhibition Alleviates Allergies Caused by Antarctic Krill Tropomyosin through Improving Th1/Th2 Imbalance and Modulating Gut Microbiota.

Antarctic krill tropomyosin (AkTM) has been shown in mice to cause IgE-mediated food allergy. The objective of this work was to investigate the role of Notch signaling in AkTM-sensitized mice, as well as to determine the changes in gut microbiota composition and short-chain fatty acids (SCFAs) in the allergic mice. An AkTM-induced food allergy mouse model was built and N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) was used as an γ-secretase inhibitor to inhibit the activation of Notch signaling. Food allergy indices, some key transcription factors, histologic alterations in the small intestine, and changes in gut microbiota composition were examined. The results showed that DAPT inhibited Notch signaling, which reduced AkTM-specific IgE, suppressed mast cell degranulation, decreased IL-4 but increased IFN-γ production, and alleviated allergic symptoms. Quantitative real-time PCR and Western blotting analyses revealed that expressions of Hes-1, Gata3, and IL-4 were down-regulated after DAPT treatment, accompanied by increases in T-bet and IFN-γ, indicating that Notch signaling was active in AkTM-sensitized mice and blocking it could reverse the Th1/Th2 imbalance. Expressions of key transcription factors revealed that Notch signaling could promote Th2 cell differentiation in sensitized mice. Furthermore, 16S rRNA sequencing results revealed that AkTM could alter the diversity and composition of gut microbiota in mice, leading to increases in inflammation-inducing bacteria such as Enterococcus and Escherichia-Shigella. Correlation analysis indicated that reduced SCFA concentrations in AkTM-allergic mice may be related to decreases in certain SCFA-producing bacteria, such as Clostridia_UCG-014. The changes in gut microbiota and SCFAs could be partially restored by DAPT treatment. Our findings showed that inhibiting Notch signaling could alleviate AkTM-induced food allergy by correcting Th1/Th2 imbalance and modulating the gut microbiota.

Metal-Free Nitrogen-Doped Mesoporous Carbons for the Mild and Selective Synthesis of Pyrroles from Nitroarenes via Cascade Reaction.

The cascade synthesis of pyrroles from nitroarenes is an attractive alternative strategy. However, metal catalysts and relatively high temperatures cover the existing reported catalytic systems for this strategy. The development of nonmetallic heterogeneous catalytic systems for the one-pot synthesis of pyrrole from nitroarenes under mild conditions is both worthwhile and challenging. Herein, we describe an exceptionally efficient method for the synthesis of N-substituted pyrroles by the reductive coupling of nitroarenes and diketones over heterogeneous metal-free catalysts under mild conditions. Nonmetallic NC-X catalysts with high activity were prepared from the pyrolysis of well-defined ligands via simple sacrificing hard template methods. Hydrazine hydrate, formic acid, and molecular hydrogen can all be used as reducing agents in the hydrogenation/Paal-Knorr reaction sequence to efficiently synthesize various N-substituted pyrroles, including drugs and bioactive molecules. The catalytic system was featured with good tolerance to sensitive functional groups and no side reactions such as dehalogenation and aromatics hydrogenation. Hammett correlation studies have shown that the electron-donating substituents are beneficial for the one-pot synthesis of N-substituted pyrroles. The results established that the outstanding performance of the catalyst is mainly attributed to the contribution of graphitic N in the catalyst as well as the promotion effect of the mesoporous structure on the reaction.

Fe single atoms encapsulated in N, P-codoped carbon nanosheets with enhanced peroxidase-like activity for colorimetric detection of methimazole.

Excessive use of antithyroid drug methimazole (MMI) in pharmaceutical samples can cause hypothyroidism and symptoms of metabolic decline. Hence, it is urgent to develop rapid, low cost and accurate colorimetric method with peroxidase-like nanozymes for determination of MMI in medical, nutrition and pharmaceutical studies. Herein, Fe single atoms were facilely encapsulated into N, P-codoped carbon nanosheets (Fe SAs/NP-CSs) by a simple pyrolysis strategy, as certified by a series of characterizations. UV-vis absorption spectroscopy was employed to illustrate the high peroxidase-mimicking activity of the resultant Fe SAs/NP-CSs nanozyme through the typical catalysis of 3,3',5,5'-tetramethylbenzidine (TMB) oxidation. The catalytic mechanism was scrutionously investigated by the fluorescence spectroscopy and electron paramagnetic resonance (EPR) tests. Additionally, the introduced MMI had the ability to reduce the oxidation of TMB (termed oxTMB) as a peroxidase inhibitor, coupled by fading the blue color. By virtue of the above findings, a visual colorimetric sensor was established for dual detection of methimazole (MMI) with a linear scope of 5-50 mM and a LOD of 1.57 mM, coupled by assay of H2O2 at a linear range of 3-50 mM. According to the irreversible oxidation of the drug, its screening with acceptable results was achieved on the sensing platform even in commercial tablets The Fe SAs/NP-CSs nanozyme holds great potential for clinical diagnosis and drug analysis.

Determination of Eugenol Residues in Fish Tissue, Transport, and Temporary Water of Aquatic Product by Gas Chromatography-Tandem Mass Spectrometry with Application of the Electrospun Nanofibrous Membrane.

Using gas chromatography-tandem mass spectrometry and electrospun nanofibrous membrane, we developed and validated a simple, rapid, and sensitive methodology for quantifying eugenol residues in fish tissue and water samples. Fish tissue extract and water samples (315 samples) collected from three southeastern China provinces (Shanghai, Zhejiang, and Fujian), originating from eight provinces of Zhejiang, Jiangsu, Shandong, Guangdong, Fujian, Anhui, Shanghai, and Jiangxi, from April 2021 to April 2023 were filtered with an electrospun nanofiber membrane, extracted with trichloromethane/n-hexane, and directly concentrated to dry after simple purification. An internal standard of p-terphenyl in n-hexane and 5-µL injection volumes of the solutions was used to analyze eugenol via internal calibration with a minimum concentration of 0.5 µg/L in water samples and 0.1 µg/kg in aquatic product samples. The highest amount of eugenol was detected in Fujian province, possibly due to the higher temperature during transportation, while the lowest amount was found in Shanghai, which mainly uses temporary fish-culture devices. This is a fast, inexpensive, and effective method for testing large quantities of fish water and meat samples.

BCL6 promotes a stem-like CD8+ T cell program in cancer via antagonizing BLIMP1.

Overcoming CD8+ T cell exhaustion is critical in cancer immunotherapy. Recently, an intratumor stem/progenitor-like CD8+ T cell (Tprog cell) population that mediates the persistence of antitumor responses has been defined, which can further develop into a terminally differentiated CD8+ T cell (Tterm cell) subpopulation with potent cytotoxic functions. Tprog cells are the main responders to immune checkpoint blockade therapies, yet how extrinsic signals via transcription factors control Tprog cell generation and persistence in tumors is unclear. Here, we found that BCL6 inhibits tumor-specific Tterm cell generation from Tprog cell downstream of TCF1. We show that Bcl6 deficiency reduced the persistence of Tprog cells, without affecting their generation, thus abrogating long-term tumor control. High-level BCL6 expression was observed in tumor-specific T cells in draining lymph nodes (LNs) and was associated with T cell exhaustion. This was observed in TOX+TCF1+ Tprog cells in both LNs and tumors. BCL6 expression in CD8+ T cells was up-regulated by TGF-ß-SMAD2 signaling but down-regulated by the IL-2-STAT5 pathway. Mechanistically, BCL6 transcriptionally repressed the expression of Tterm cell-associated genes and induced those of Tprog cell-related genes, in a manner antagonistic to BLIMP1. Prdm1 deficiency also promoted the Tprog cell program and greatly improved the efficacy of anti-PD-1 therapy. Thus, we identified the TGF-ß-BCL6 and IL-2-BLIMP1 antagonistic pathways in regulation of antitumor CD8+ T cells, which may benefit the development of long-lasting and effective cancer immunotherapy.

Advances in mRNA-Based Cancer Vaccines.

Cancer is a leading cause of death worldwide, accounting for millions of deaths every year. Immunotherapy is a groundbreaking approach for treating cancer through harnessing the power of the immune system to target and eliminate cancer cells. Cancer vaccines, one immunotherapy approach, have shown promise in preclinical settings, but researchers have struggled to reproduce these results in clinical settings. However, with the maturity of mRNA technology and its success in tackling the recent coronavirus disease 2019 (COVID-19) pandemic, cancer vaccines are expected to regain attention. In this review, we focused on the recent progress made in mRNA-based cancer vaccines over the past five years. The mechanism of action of mRNA vaccines, advancements in neoantigen discovery, adjuvant identification, and delivery materials are summarized and reviewed. In addition, we also provide a detailed overview of current clinical trials involving mRNA cancer vaccines. Lastly, we offer an insight into future considerations for the application of mRNA vaccines in cancer immunotherapy. This review will help researchers to understand the advances in mRNA-based cancer vaccines and explore new dimensions for potential immunotherapy approaches.

Safety, tolerability, and immunogenicity of a CpG/Alum adjuvanted SARS-CoV-2 recombinant protein vaccine (ZR202-CoV) in healthy adults: Preliminary report of a phase 1, randomized, double-blind, placebo-controlled, dose-escalation trial.

This was a phase 1 dose-escalation study of ZR202-CoV, a recombinant protein vaccine candidate containing a pre-fusion format of the spike (S)-protein (S-trimer) combined with the dual-adjuvant system of Alum/CpG. A total of 230 participants were screened and 72 healthy adults aged 18-59 years were enrolled and randomized to receive two doses at a 28-day interval of three different ZR202-CoV formulations or normal saline. We assessed the safety for 28 days after each vaccination and collected blood samples for immunogenicity evaluation. All formulations of ZR202-CoV were well-tolerated, with no observed solicited adverse events ≥ Grade 3 within 7 days after vaccination. No unsolicited adverse events ≥ Grade 3, or serious adverse events related to vaccination occurred as determined by the investigator. After the first dose, detectable immune responses were observed in all subjects. All subjects that received ZR202-CoV seroconverted at 14 days after the second dose by S-binding IgG antibody, pseudovirus and live-virus based neutralizing antibody assays. S-binding response (GMCs: 2708.7 ~ 4050.0 BAU/mL) and neutralizing activity by pseudovirus (GMCs: 363.1 ~ 627.0 IU/mL) and live virus SARS-CoV-2 (GMT: 101.7 ~ 175.0) peaked at 14 days after the second dose of ZR202-CoV. The magnitudes of immune responses compared favorably with COVID-19 vaccines with reported protective efficacy.

[Comparison of therapeutic effects of arthroscopic popliteal cyst internal drainage and capsular wall resection].

OBJECTIVE: To investigate efficacy between arthroscopic popliteal cyst drainage and arthroscopic popliteal cyst resection. METHODS: From January 2013 to June 2021, 54 patients with popliteal cyst (Rausching-Lindgren gradeⅠto Ⅲ) were treated with arthroscopic surgery. There were 24 males and 30 females. The age ranged from 44 to 72 years old, with a mean of (62.67±6.08) years old. The course of the disease ranged from 1 to 72 months, with a mean of(15±14) months. Twenty-four patients (group A) were underwent arthroscopic internal drainage of popliteal cyst. Thirty patients (group B) were underwent arthroscopic resection of popliteal cyst. Preoperative main symptoms included knee pain, swelling, walking pain, popliteal swelling, popliteal mass and so on. After 1, 3, 6 months and 1, 2 years of surgery, routine outpatient follow-up was conducted to observe and compare the surgical time, bleeding volume, preoperative and postoperative visual analog scale (VAS), knee Lysholm score, and complications between two groups. RESULTS: All incisions healed at one stage after operation. All 54 patients were followed up, and the duration ranged from 6 months to 2 years, with an average of (13.89±4.29) months. There was no intraoperative vascular or nerve injury. Operation time and intraoperative blood loss of the two groups:group A of (62.08±9.55) min and (8.00±1.69) ml, group B of (69.50±6.99) min and (8.70±2.00) ml. Popliteal pain, swelling, limitation of flexion and extension were significantly relieved after operation. VAS before and one month after operation between two groups:group A of 5.38±1.21 and 2.63±0.71, group B of 5.60±1.26 and 2.80±0.81. Lysholm scores of knee joint before and 6 months after operation:group A of 62.59±4.99 and 89.74±2.90, group B of 63.87±3.23 and 89.02±2.35. Knee joint function improved significantly in both groups. In group A, 4 cases had popliteal cyst at 3 months after operation, and 2 cases had small isolated cyst at 1 year after operation. There was no recurrence of cyst in group B. CONCLUSION: The results between two arthroscopic treatments of popliteal cyst are satisfactory, and there is no significant difference in the amount of blood loss, safety, postoperative pain VAS score and knee function recovery. It is suggested that arthroscopic resection of the cyst wall should be performed when the technique is mature, especially for large cysts and septal cysts.

Acetyl-coenzyme A acetyltransferase 1 promotes brown adipogenesis by activating the AMPK-PGC1α signaling pathway.

Brown adipose tissue (BAT) is thermogenic, expressing high levels of uncoupling protein-1 to convert nutrient energy to heat energy, bypassing ATP synthesis. BAT is a promising therapeutic target for treatment of obesity and type 2 diabetes since it converts fatty acids into heat but mechanisms controlling brown adipogenesis remain unclear. Knockdown of acetyl-Coenzyme A acetyltransferase 1 (ACAT1) in C3H10T1/2 cells suppressed brown adipocyte maturation during the current study and ACAT1 overexpression promoted brown adipocyte maturation. The downstream target of AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor gamma coactivator-1-α (PGC1α), was involved in the action of ACAT1 on brown adipocyte maturation. ACAT1 overexpression enhanced AMPK phosphorylation and promoted PGC1α expression. It is suggested that ACAT1 promotes brown adipocyte maturation by activating the AMPK-PGC1α signaling pathway.

Recent Advances in Sources, Migration, Public Health, and Surveillance of Bisphenol A and Its Structural Analogs in Canned Foods.

The occurrence of bisphenol A (BPA) and its structural analogs, known as endocrine disruptors is widely reported. Consumers could be exposed to these chemicals through canned foods, leading to health risks. Considerable advances have occurred in the pathogenic mechanism, migration law, and analytical methodologies for these compounds in canned foods. However, the confusion and controversies on sources, migration, and health impacts have plagued researchers. This review aimed to provide insights and perspectives on sources, migration, effects on human health, and surveillance of these chemicals in canned food products. Current trends in the determination of BPA and its structural analogs have focused on mass spectroscopy and electrochemical sensor techniques. Several factors, including pH, time, temperature, and volume of the headspace in canned foods, could affect the migration of the chemicals. Moreover, it is necessary to quantify the proportion of them originating from the can material used in canned product manufacturing. In addition, adverse reaction research about exposure to low doses and combined exposure with other food contaminants will be required. We strongly believe that the information presented in this paper will assist in highlighting the research needs on these chemicals in canned foods for future risk evaluations.

Low level TGF-ß1-treated Umbilical mesenchymal stem cells attenuates microgliosis and neuropathic pain in chronic constriction injury by exosomes/lncRNA UCA1／miR-96-5p／FOXO3a.

Neuropathic pain is a chronic pain state that usually caused by injuries in peripheral or central nerve. Inhibition of spinal microglial response is a promising treatment of neuropathic pain caused by peripheral nerve injury. In recent years, mesenchymal stem cells (MSCs) that characterized with multipotent ability have been widely studied for disease treatment. TGF-ß1 is a well-known regulatory cytokine that participate in the response to cell stress and is closely correlated with the function of nerve system as well as MSC differentiation. This work aimed to determine the effects of exosomes that extracted from TGF-ß1-induced umbilical mesenchymal stem cells (hUCSMCs) on the neuropathic pain. In this work, we established a rat model of chronic constriction injury (CCI) of the sciatic nerve and LPS-induced microglia cell model. The hUCSMCs cell surface biomarker was identified by flow cytometry. Exosomes that extracted from TGF-ß1-treated hUCSMCs were characterized by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA) and used for treatment. We observed that TGF-ß1 upregulates the level of lncRNA UCA1 (UCA1) in hUCMSC-derived exosomes. Treatment with exosomal lncRNA UCA1 (UCA1) alleviated the neuropathic pain, microgliosis, and production of inflammatory mediator both in vivo and in vitro. UCA1 directly interact with the miR-96-5p, and the miR-96-5p acts as sponge of FOXO3a. Knockdown of UCA1 upregulated the level of miR-96-5p and downregulated the FOXO3a expression, which could be recovered by inhibition of miR-96-5p. In summary, the TGF-ß1-stimulated exosomal UCA1 from hUCMSCs alleviates the neuropathic pain and microgliosis. These findings may provide novel evidence for treatment of neuropathic pain caused by chronic constriction injury.

Study on the Sensitization and Antigenic Epitopes of Tropomyosin from Antarctic Krill (Euphausia superba).

Antarctic krill (Euphausia superba), a shrimp-like marine crustacean, has become a beneficial source of high-quality animal protein. Meanwhile, a special focus has been placed on its potential sensitization issue. In this study, a 35 kDa protein was purified and identified to be Antarctic krill tropomyosin (AkTM) by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The purified TM showed a strong IgE-binding capacity to shrimp/crab-allergic patients' sera, indicating that TM is the primary allergen in Antarctic krill. Simulated gastrointestinal digestion revealed that the digestion stability of TM to pepsin was higher than that to trypsin. The strong degranulation triggered by TM in RBL-2H3 cells suggested that AkTM has a strong sensitization capacity. The TM-sensitized BALB/c mice displayed severe anaphylactic symptoms; high levels of TM-specific IgE, sIgG1, and histamine; and increased IL-4, indicating that AkTM could provoke IgE-mediated allergic reactions. Bioinformatics prediction, indirect competition ELISA, and mast cell degranulation assay were used to map the antigenic epitopes of AkTM. Finally, nine peptides of T43-58, T88-101, T111-125, T133-143, T144-155, T183-197, T223-236, T249-261, and T263-281 were identified as the linear epitopes of AkTM. The findings may help us develop efficient food processing techniques to reduce krill allergy and gain a deeper comprehension of the allergenicity of krill allergens.

Screening and Identification of Goat-Milk-Derived Lactic Acid Bacteria with Bacteriocin-like Activity and Probiotic Potentials.

In the current study, we screened 46 isolates of lactic acid bacteria (LAB) derived from goat milk for bacteriocin producers that can inhibit common foodborne pathogens (Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus). The following three strains that showed antimicrobial activity against all indicators were identified: Enterococcus faecalis DH9003 and DH9012, and Lactococcus lactis DH9011. Their antimicrobial products exhibited typical bacteriocin characteristics, such as heat stability and proteinase nature. The bacteriostatic activity of concentrated bacteriocins produced by these LAB was observed at low concentrations (half-minimum inhibitory concentration [MIC50] and 4MIC50), whereas complete inhibition activity against Listeria monocytogenes was detected at high concentrations (16MIC50) of the two Enterococcus faecalis strains (DH9003 and DH9012). Furthermore, the probiotic potentials of the three strains were investigated and described. The results revealed that none of the strains had hemolytic activity, whereas all: were sensitive to ampicillin (50 mg/mL) and streptomycin sulfate (100 mg/mL); were resistant to bile, artificial simulated intestinal tract, and gastric juice at different pH levels (2.5, 3.0, 3.5); and had ß-galactosidase activity. Furthermore, all strains exhibited an auto-aggregating phenotype, with self-aggregation ranging from 30% to 55%. DH9003 and DH9012 co-aggregated well with Listeria monocytogenes and Escherichia coli (52.6% and 63.2%, 68.5% and 57.6%, respectively), whereas DH9011 co-aggregated poorly with Listeria monocytogenes (15.6%) and did not co-aggregate with Escherichia coli. Furthermore, our results showed that all three isolates exhibited strong antibacterial activity, tolerance to bile and simulated gastrointestinal environments, adhesion capability, and safety. Finally, DH9003 was selected and used for gavage in rats. By observing the pathological characteristics of rat intestinal and liver tissue sections, DH9003 showed no harmful effects on the intestine and liver of rats, but rather resulted in a denser and longer intestinal mucosa, as well as improving the intestinal mucosa of rats. Considering their substantial prospective applications, we concluded that these three isolates are potential probiotic candidates.