期刊：
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY,2025年15:1585293 ISSN：2235-2988
通讯作者：
Chen, Jiayi;Wang, KJ
作者机构：
[Chen, Jiayi; Liu, Huixin; Wang, Kaijun] Changsha Med Univ, Hunan Prov Key Lab Tradit Chinese Med Agr Biogenom, Changsha, Peoples R China.;[Liu, Huixin] Sichuan Acad Chinese Med Sci, Anim Expt Ctr, Chengdu, Peoples R China.;[Liu, Huixin; Wang, Chenchen; Wei, Xiaofang; He, Yang] Guangxi Univ, Coll Anim Sci & Technol, Key Lab Anim Breeding Dis Control & Prevent, Nanning, Peoples R China.;[Wang, Kaijun] Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.;[Liu, Huixin] Sichuan Inst Tradit Chinese Med, Expt Anim Ctr, Chengdu, Sichuan, Peoples R China.
通讯机构：
[Chen, JY; Wang, KJ ] C;Changsha Med Univ, Hunan Prov Key Lab Tradit Chinese Med Agr Biogenom, Changsha, Peoples R China.;Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.
关键词：
infectious bronchitis virus;traditional Chinese medicine inheritance computing platform;vitro and in vivo experiments;liquid chromatography-mass spectrometry;network pharmacology
摘要：
INTRODUCTION: Infectious bronchitis virus (IBV) imposes severe economic burdens on the poultry industry, and current treatments face challenges in efficacy and sustainability, necessitating the development of novel therapeutic strategies. To address this, this study employed the Traditional Chinese Medicine Inheritance Computing Platform (TCMICS) to collect clinical prescriptions for IBV treatment, based on which two optimized versions of the traditional Chinese medicine Maxing Shigan Decoction (MXSG), namely MXSG-mix1 and MXSG-mix2, were designed. In vitro cell culture and in vivo chicken model experiments were then carried out, including egg testing, clinical symptom observation, immune function analysis, and viral load quantification, to assess the antiviral activity of the optimized formulations. METHODS: To explore the underlying mechanisms, liquid chromatography-mass spectrometry (LC-MS) was combined with network pharmacology to identify 28 active compounds in MXSG-mix and 47 key genes involved in viral replication, inflammation, and apoptosis pathways. Furthermore, molecular docking and RT-qPCR were performed, which confirmed that MXSG-mix downregulated BCL2 expression and interacted with AKT1 and CASP3, thus inhibiting IBV-induced cell apoptosis. RESULTS AND DISCUSSION: The results showed that both MXSG-mix1 and MXSG-mix2 exhibited superior anti-IBV activity compared to traditional MXSG, effectively reducing viral load and improving immune responses in vivo. In conclusion, integrating TCMICS, LC-MS, and network pharmacology offers a novel paradigm for developing veterinary TCM formulations. The optimized MXSG-mix shows potential as an effective, multi-target therapeutic against IBV, providing valuable insights for future anti-viral drug development in poultry medicine.
摘要：
Acrorus tatarinowii Schott (ATS), a traditional medicinal herb with neuroprotective potential, requires optimized harvest timing to maximize its bioactive efficacy. This study integrated HS-SPME-GC-MS, network pharmacology, and molecular dynamics simulations to evaluate seasonal variations in volatile components and their therapeutic relevance for neurodegenerative diseases. Monthly samples (March – December) revealed that the essential oil yield peaked in November (2.6 %) and declined by December. Phenylpropanoids dominated the volatile profile (e.g., β-Asarone, 19.32–35.81 %), showing progressive accumulation, while sesquiterpenes peaked in August. Network pharmacology identified 37 bioactive compounds targeting neurodegenerative pathways, with ESR1, SRC, and MAPK3 as core targets. Molecular docking highlighted β-eudesmol (Ki = 8.2 μM), (+)-tau-muurolol, and β-Asarone as high-affinity ligands for ESR1. Dynamics simulations confirmed stable binding (RMSD <0.4 nm) and favorable free energy (ΔG ≤-90.17 kJ/mol). The seasonal analysis, which aligned with pharmacopoeial guidelines, indicated that autumn (optimal essential oil yield) and winter (high β-Asarone content) are the ideal harvest periods. This multidisciplinary approach offers a scientific foundation for industrial-scale ATS cultivation, ensuring consistent quality for neurotherapeutic applications.
Acrorus tatarinowii Schott (ATS), a traditional medicinal herb with neuroprotective potential, requires optimized harvest timing to maximize its bioactive efficacy. This study integrated HS-SPME-GC-MS, network pharmacology, and molecular dynamics simulations to evaluate seasonal variations in volatile components and their therapeutic relevance for neurodegenerative diseases. Monthly samples (March – December) revealed that the essential oil yield peaked in November (2.6 %) and declined by December. Phenylpropanoids dominated the volatile profile (e.g., β-Asarone, 19.32–35.81 %), showing progressive accumulation, while sesquiterpenes peaked in August. Network pharmacology identified 37 bioactive compounds targeting neurodegenerative pathways, with ESR1, SRC, and MAPK3 as core targets. Molecular docking highlighted β-eudesmol (Ki = 8.2 μM), (+)-tau-muurolol, and β-Asarone as high-affinity ligands for ESR1. Dynamics simulations confirmed stable binding (RMSD <0.4 nm) and favorable free energy (ΔG ≤-90.17 kJ/mol). The seasonal analysis, which aligned with pharmacopoeial guidelines, indicated that autumn (optimal essential oil yield) and winter (high β-Asarone content) are the ideal harvest periods. This multidisciplinary approach offers a scientific foundation for industrial-scale ATS cultivation, ensuring consistent quality for neurotherapeutic applications.
通讯机构：
[Feng, ZX; Yu, YF ] N;Nanjing Agr Univ, Coll Vet Med, Nanjing 210095, Peoples R China.
关键词：
ERA;Mycoplasma contamination;PCR;qPCR
摘要：
Mycoplasma contamination poses a persistent challenge in cell culture systems and the production of cell-derived biological products, including vaccines and therapeutic drugs. Current detection methods suffer from several limitations: they risk false-negative results due to incomplete species coverage, exhibit high detection limits in molecular assays, and prove time-consuming while lacking sensitivity for certain fastidious mycoplasma species that grow poorly in vitro , as outlined in pharmacopeial testing standards. To address these issues, we developed three improved detection methods—PCR, enzymatic recombinase amplification (ERA), and quantitative PCR (qPCR)—using universal primer pairs targeting conserved regions across 143 mycoplasma species. All three methods demonstrated exceptional specificity, accurately identifying 16 different Mycoplasma species (including those specified in the European Pharmacopoeia [2021] and Japanese Pharmacopoeia [JP16]) while showing no cross-reactivity with common cell culture contaminants (bacteria, viruses, or fungi). Sensitivity testing using a Spiroplasma 16S-23S spacer fragment plasmid revealed detection limits of 10ˆ1 copies for PCR, 10ˆ0 copies for ERA, and an impressive 10ˆ-1 copies for qPCR. Validation studies showed 100% agreement with pharmacopeial gold-standard methods, with our novel methods achieving 17% (PCR), 36.8% (ERA), and 40.6% (qPCR) higher detection rates than conventional approaches and outperforming comparable commercial products. Notably, all three methods significantly reduced testing time to just 1-2 hours. Our results demonstrate substantial improvements in specificity, sensitivity, and detectable species range compared to existing methods. Among these approaches, qPCR emerges as particularly promising due to its superior sensitivity and rapid turnaround, making it ideally suited for quality control in biological product manufacturing and research applications. This advancement represents a significant step forward in mycoplasma contamination monitoring for the biotechnology and pharmaceutical industries.
Mycoplasma contamination poses a persistent challenge in cell culture systems and the production of cell-derived biological products, including vaccines and therapeutic drugs. Current detection methods suffer from several limitations: they risk false-negative results due to incomplete species coverage, exhibit high detection limits in molecular assays, and prove time-consuming while lacking sensitivity for certain fastidious mycoplasma species that grow poorly in vitro , as outlined in pharmacopeial testing standards. To address these issues, we developed three improved detection methods—PCR, enzymatic recombinase amplification (ERA), and quantitative PCR (qPCR)—using universal primer pairs targeting conserved regions across 143 mycoplasma species. All three methods demonstrated exceptional specificity, accurately identifying 16 different Mycoplasma species (including those specified in the European Pharmacopoeia [2021] and Japanese Pharmacopoeia [JP16]) while showing no cross-reactivity with common cell culture contaminants (bacteria, viruses, or fungi). Sensitivity testing using a Spiroplasma 16S-23S spacer fragment plasmid revealed detection limits of 10ˆ1 copies for PCR, 10ˆ0 copies for ERA, and an impressive 10ˆ-1 copies for qPCR. Validation studies showed 100% agreement with pharmacopeial gold-standard methods, with our novel methods achieving 17% (PCR), 36.8% (ERA), and 40.6% (qPCR) higher detection rates than conventional approaches and outperforming comparable commercial products. Notably, all three methods significantly reduced testing time to just 1-2 hours. Our results demonstrate substantial improvements in specificity, sensitivity, and detectable species range compared to existing methods. Among these approaches, qPCR emerges as particularly promising due to its superior sensitivity and rapid turnaround, making it ideally suited for quality control in biological product manufacturing and research applications. This advancement represents a significant step forward in mycoplasma contamination monitoring for the biotechnology and pharmaceutical industries.
通讯机构：
[Ge, M ] H;Hunan Agr Univ, Coll Vet Med, Changsha 410125, Peoples R China.
关键词：
Porcine circovirus 2;ELISpot;Memory B cells;Immune memory;Vaccine
摘要：
Porcine Circovirus 2 (PCV2) vaccination plays a crucial role in preventing porcine circovirus-associated disease (PCVAD). Nevertheless, pig farms face significant challenges in evaluating vaccination efficacy due to the inability of PCV2 vaccines to achieve sterilizing immunity and the variability among vaccine manufacturers. These challenges are further compounded by the limitations of conventional antibody detection methods, which fail to distinguish between maternally-derived antibodies (MDAs) and vaccine-induced antibodies. The accurate evaluation and selection of PCV2 vaccines is critical for the swine industry. The present study aimed to develop an Enzyme-linked immunospot (ELISpot) assay for directly detecting PCV2-specific memory B cells. This approach was used to assess the presence of PCV2-specific memory B cells in piglets with high levels of MDA vaccinated with different PCV2 vaccines, thus enabling the evaluation of vaccine immunogenicity at the cellular level. Furthermore, antibody levels and the viremia status were analyzed using Enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qPCR) respectively to provide a comprehensive assessment of the ELISpot assay potential for evaluating the vaccine immunogenicity of PCV2 vaccines. The findings revealed that the optimal conditions for the developed ELISpot assay included stimulation with R848 at a final concentration of 1 µg·mL⁻¹ for three days, a PCV2 Cap protein coating concentration of 1.25 µg·mL⁻¹, a biotinylated goat anti-pig IgG antibody concentration of 5 µg·mL⁻¹, and an HRP-streptavidin concentration of 0.25 µg·mL⁻¹. In high MDA piglets immunized with different vaccines, serum antibody detection showed that PCV2 antibody levels declined continuously over time in all vaccinated and saline-injected control groups, demonstrating similar trends. In contrast, ELISpot analysis demonstrated a significant increase in PCV2-specific memory B cell levels in all three vaccinated groups compared to the saline-injected group. Among the vaccines tested, Vaccine A induced the highest levels of specific memory B cells, followed by Vaccine B. This was consistent with the lower PCV2 infection rates and viremia levels observed in Vaccine A and Vaccine B groups, compared to Vaccine C and saline-injected control groups. We established an ELISpot assay to quantify PCV2-specific memory B cells, revealing that vaccinated piglets with high MDA levels developed robust memory B cell responses. However, levels of PCV2 IgG antibodies in vaccinated piglets remained statistically indistinguishable from control piglets. These findings demonstrate that ELISpot-based profiling of PCV2-specific memory B cells overcomes the confounding effects of MDA in vaccine efficacy assessments. This approach reliably reflects the humoral immune response induced by vaccination and its relevance in combating natural PCV2 infection, providing valuable guidance for preventing and controlling PCVAD.
摘要：
Gamma-aminobutyric acid receptors (GABARs) primarily function by suppressing inflammatory responses, modulating neuronal excitability, and maintaining intracellular homeostasis, whereas N-methyl-D-aspartate receptors (NMDARs) play a key role in mediating pathological processes through the regulation of excitatory neurotransmission and immune responses. Viral infections have the capacity to modify the expression and functionality of these receptors, either directly or indirectly, thereby contributing to dysregulation within the neurological and immune systems and triggering a range of disease states. This review offers a comprehensive analysis of the mechanisms through which various viral infections interact with GABARs and NMDARs, emphasizing the possible intricate roles these receptors play in viral pathogenesis. Additionally, it underscores their potential as therapeutic targets for antiviral interventions, particularly in addressing immune dysregulation and neurological disorders.
通讯机构：
[Ren, G ; Ren, G; Zhang, Y ] N;[Zhao, YX ; Zhao, SH] Y;Northwest A&F Univ, Minist Agr, Key Lab Anim Biotechnol, Xianyang 712100, Peoples R China.;Yazhouwan Natl Lab, Huanjin Rd, Sanya 572024, Hainan, Peoples R China.;Northwest A&F Univ, Coll Anim Sci & Technol, Yangling 712100, Peoples R China.
关键词：
3D interaction;RNA Pol II;SPDs;ZGA pace;interspecies comparison;zygotic genome activation
摘要：
Zygotic genome activation (ZGA) occurs at distinct stages across mammals, with mice initiating ZGA at the 2-cell stage and bovines and humans activating the process in the 4- to 8-cell stages. RNA polymerase II (RNA Pol II) gradually initiates ZGA in mice, but regulation in late-ZGA species remains unclear. Here, RNA Pol II profiling in bovine embryos identified strong intergenic clusters that boost minor ZGA gene expression via chromatin interactions and are named super RNA Pol II domains (SPDs). CRISPRi perturbation of SPDs in bovine embryos decreases the expression of minor ZGA genes, whereas the knockdown of these genes disrupts major ZGA and embryogenesis. Rapid enhancement of minor ZGA genes also occurs in human embryos. Alternatively, mouse and porcine oocytes precociously express these minor ZGA genes without SPDs. Thus, SPDs appear to be an adaptation in bovine embryos, promoting minor ZGA gene expression to comparable levels as early-ZGA species, illuminating species-specific regulation of ZGA timing.
Zygotic genome activation (ZGA) occurs at distinct stages across mammals, with mice initiating ZGA at the 2-cell stage and bovines and humans activating the process in the 4- to 8-cell stages. RNA polymerase II (RNA Pol II) gradually initiates ZGA in mice, but regulation in late-ZGA species remains unclear. Here, RNA Pol II profiling in bovine embryos identified strong intergenic clusters that boost minor ZGA gene expression via chromatin interactions and are named super RNA Pol II domains (SPDs). CRISPRi perturbation of SPDs in bovine embryos decreases the expression of minor ZGA genes, whereas the knockdown of these genes disrupts major ZGA and embryogenesis. Rapid enhancement of minor ZGA genes also occurs in human embryos. Alternatively, mouse and porcine oocytes precociously express these minor ZGA genes without SPDs. Thus, SPDs appear to be an adaptation in bovine embryos, promoting minor ZGA gene expression to comparable levels as early-ZGA species, illuminating species-specific regulation of ZGA timing.
作者机构：
[Wu, X; Wu, Xiang; Fu, Yi-Tian] Cent South Univ, Xiangya Sch Basic Med Sci, Dept Parasitol, Changsha 410013, Hunan, Peoples R China.;[Fu, Yi-Tian; Deng, Yuan-Ping; Liu, Guo-Hua; Peng, Yan-Yan; Wang, Hui-Mei] Hunan Agr Univ, Coll Vet Med, Res Ctr Parasites & Vectors, Changsha 410128, Hunan, Peoples R China.;[Xie, Yue] Sichuan Agr Univ, Coll Vet Med, Dept Parasitol, Chengdu 611130, Sichuan, Peoples R China.
通讯机构：
[Wu, X ; Fu, YT] C;[Fu, YT ] H;Cent South Univ, Xiangya Sch Basic Med Sci, Dept Parasitol, Changsha 410013, Hunan, Peoples R China.;Hunan Agr Univ, Coll Vet Med, Res Ctr Parasites & Vectors, Changsha 410128, Hunan, Peoples R China.
关键词：
Head lice;Prevalence;Impact factor;Schoolchildren;Duplex PCR
摘要：
BACKGROUND: Head lice (Pediculus humanus capitis) infestation is a worldwide public health concern, especially in school-aged children. However, its main impact factors and genetic characteristics remain poorly understood in China. Hence, the purpose of the study was to explore the precise association between multiple impact factors and head lice infestation, characterize the genetic variation of the head lice, and develop a sensitive and specific mitochondrial (mt) DNA duplex polymerase chain reaction (PCR) for accurately distinguishing clades A and B. METHODS: A cross-sectional study was conducted in Hunan Province, central China from January 2018 to July 2023. A total of 9254 schoolchildren from 48 primary schools in each administrative region were examined for head lice. Impact factors for infestation were analyzed using the data collected by a questionnaire. The mt cytb gene sequences of head lice collected in the current study were used for sequence analysis, then were added to the global pool to conduct the phylogenetic analyses. Primers designed on the basis of this gene sequence were used in duplex PCR to diagnose head lice clades A and B by amplicon size. RESULTS: Head lice infestation was found in 93.8% (45/48) of the primary schools included in the study. Overall, 6.8% (630/9254) of the examined schoolchildren harbored head lice, with 94.6% (596/630) being girls. A total of 2132 adult head lice were collected from 630 infested cases. The impact factors for head lice infestation included gender, school location, family situation, per capita income, study mode, and hair washing per week (p < 0.01). However, season and age were not considered as impact factors for head lice prevalence (p > 0.05). Phylogenetic analysis based on mt cytb gene sequences showed that head lice are classified into two clades (A and B), with clade B being more dominant in Hunan Province, central China. The newly developed duplex PCR was able to differentiate clades A from B in China with 100% sensitivity and specificity. CONCLUSIONS: Our findings revealed that head lice infestation is mostly associated with poverty and poor hygiene in Hunan Province, central China. It is crucial to consider the simultaneous surveillance of head lice infestation in schoolchildren in regions with low level of socioeconomic status; however, datasets from other provinces are warranted to confirm the findings. It further showed that clades A and B are common in central China and that the latter has emerged and become the dominant one.
摘要：
Aeromonas hydrophila is an important zoonotic bacterium that is related to multiple diseases in humans, terrestrial animals, and aquatic animals. Bacterial septicemia caused by A. hydrophila often results in high mortality and severe economic losses. Antibiotics, the major approach to dealing with bacterial infections, are limited due to the occurrence of antibiotic resistance. Anti-virulence strategies provide a promising approach to combat resistant bacterial infections. Here, growth curves, hemolysis, biofilm formation, and animal studies were performed to investigate the effect of fisetin against A. hydrophila. Moreover, RNA-seq technology was employed to determine the potent mechanism of fisetin. The results showed that fisetin could dose-dependently reduce the hemolytic activities mediated by aerolysin and hinder biofilm formation. Animal studies showed that treatment with 50 mg/kg fisetin could remarkably reduce the mortality to 40% in the infected group compared with fish in the fisetin-free group. Further, transcriptome analysis demonstrated that there were 565 differentially expressed genes (DEGs) after treatment with 16 μg/mL fisetin. Fisetin significantly impacted the pathways related to oxidative phosphorylation, the citrate cycle, and virulence factor regulation. Furthermore, 159 virulence-related genes were influenced after fisetin treatment. Collectively, these findings revealed that fisetin could mitigate the pathogenicity of A. hydrophila by affecting oxidative phosphorylation and the citrate cycle pathway as well as inhibiting the production of virulence factors. The study not only identified a powerful substance for managing A. hydrophila-associated diseases in aquaculture but also clarified the mechanism of plant medicines in controlling diseases caused by bacterial pathogens.
通讯机构：
[Liu, M ; Huang, P ] H;[Huang, P] Y;Hunan Agr Univ, Coll Anim Sci & Technol, Changsha 410128, Peoples R China.;Yuelushan Lab, Changsha 410000, Peoples R China.;Hunan Agr Univ, Coll Vet, Changsha 410128, Hunan, Peoples R China.
摘要：
The perinatal period (from late gestation to the neonatal stage) in ruminants is a critical phase for fetal organ maturation, where ecological succession of gastrointestinal microbial communities significantly impacts livestock production efficiency. However, research remains insufficient regarding the distribution patterns and functional annotation of microbial communities across different gastrointestinal compartments during this period. This study characterized early microbiota dynamics in Hutianshi Goats using 16S rRNA sequencing (4 fetal goats at 90 ± 10 gestational days) and metagenomics (3 7-day-old goatkids). The fetal goat group generated 852,694 valid reads, yielding 688,277 high-quality reads after chimera removal for downstream analysis. The 7-day-old goat kids group produced 1,081,588,182 final valid reads, after data processing and assembly, 8,561,345 contigs were generated. Gene prediction identified 6,095,352 genes. Multi-database annotations (NR, KEGG, CAZy, etc.) revealed functional potential and antimicrobial resistance traits. The public release of this dataset facilitates academic understanding of microbial community dynamics and host-microbe interactions during this developmental stage, providing both theoretical foundations and data resources for ruminant developmental biology and precision breeding regulation.
摘要：
Gelsemium elegans a plant of the Loganiaceae family, is highly toxic and contains many alkaloids of potential pharmacological value. Koumine, is the most abundant component of G. elegans , and belongs to monoterpene indole alkaloids and is valued in medical research as it has important anti-inflammatory and anxiolytic properties. The biosynthesis pathway of this compound has been little studied, and is poorly understood, limiting the ability to improve koumine production via breeding or synthetic biology. To investigate G. elegans biosynthesis of koumine, its genomes was sequenced and assembled (331.82 Mb) and a comprehensive transcriptome cDNA library from different tissues and hormone treatments constructed and sequenced using PacBio (10.9 Gb subreads). Using liquid chromatography-mass spectrometry techniques, we identified 29 alkaloids in extracts of G. elegans and analyzed their synthesis and accumulation in different tissues and this data was compared to the transcriptomic data to identify 20 candidate genes likely to be involved in the synthesis of koumine. Finally, a preliminary validation of the functions of two candidate genes GeLAMT and GeTDC were performed and found that both proteins catalyze the production of products in koumine biosynthesis. This data provides a rich molecular resource for the study of G. elegans , as well as the first functional validation of genes in G. elegans , that will help to inform further MIA biosynthetic pathway studies.
Gelsemium elegans a plant of the Loganiaceae family, is highly toxic and contains many alkaloids of potential pharmacological value. Koumine, is the most abundant component of G. elegans , and belongs to monoterpene indole alkaloids and is valued in medical research as it has important anti-inflammatory and anxiolytic properties. The biosynthesis pathway of this compound has been little studied, and is poorly understood, limiting the ability to improve koumine production via breeding or synthetic biology. To investigate G. elegans biosynthesis of koumine, its genomes was sequenced and assembled (331.82 Mb) and a comprehensive transcriptome cDNA library from different tissues and hormone treatments constructed and sequenced using PacBio (10.9 Gb subreads). Using liquid chromatography-mass spectrometry techniques, we identified 29 alkaloids in extracts of G. elegans and analyzed their synthesis and accumulation in different tissues and this data was compared to the transcriptomic data to identify 20 candidate genes likely to be involved in the synthesis of koumine. Finally, a preliminary validation of the functions of two candidate genes GeLAMT and GeTDC were performed and found that both proteins catalyze the production of products in koumine biosynthesis. This data provides a rich molecular resource for the study of G. elegans , as well as the first functional validation of genes in G. elegans , that will help to inform further MIA biosynthetic pathway studies.
摘要：
BACKGROUND: Gelsemium elegans (G. elegans) is widely recognized as one of the most toxic plants globally, particularly harmful to humans. Some reports indicate that it is non-toxic to pigs and even has a growth-promoting effect; however, the underlying reasons for this paradox remain unclear. METHODS: Gelsenicine is the main toxic component of G. elegans. This study characterized gelsenicine-induced toxicity using electrophysiological recordings, molecular dynamic simulations, c-Fos immunostaining, and multi-omics technologies. Additionally, we conducted a comprehensive analysis comparing the toxic effects of gelsenicine across various animal species through examinations of tissue distribution, blood gas analysis, metabonomics, and behavioral tests. RESULTS: We demonstrated that gelsenicine-induced hypoxia leads to respiratory depression in mice by enhancing the effect of gamma-aminobutyric acid (GABA) on GABA receptors (GABARs). Glycine significantly ameliorated hypoxia and improved the survival of gelsenicine-poisoned mice. Under gelsenicine-induced hypoxic conditions, N-methyl-D-aspartate (NMDA) receptor function and mitochondrial energy metabolism processes were perturbed, resulting in neuronal excitotoxicity. Finally, we confirmed that pigs could tolerate hypoxia and were resistant to gelsenicine toxicity due to high concentrations of circulating glycine and low levels of NMDA receptors (NMDARs) in the hippocampus. CONCLUSIONS: These findings suggest that hypoxic protection should be considered as a potential therapeutic strategy for gelsenicine poisoning. Our study contributes to preventing potential risks posed by G. elegans poisoning to human and animal health.
作者机构：
[Liyun Yuan] College of Agronomy, Xiangyang Polytechnic, Xiangyang, China;[You Huang; Chaoyang Ma; Lijuan Zhu; Li Kong; Chunlin Huang; Wenjiang Yang; Jiayu He; Mingqi Yang; Lin Huang; Jine Yi] Hunan Engineering Research Center of Livestock and Poultry Health Care, College of Veterinary Medicine, Hunan Agricultural University, Changsha, China
通讯机构：
[Jine Yi] H;Hunan Engineering Research Center of Livestock and Poultry Health Care, College of Veterinary Medicine, Hunan Agricultural University, Changsha, China
摘要：
As a conventional immunosuppressive drug, cyclophosphamide (CYP) exhibits strong hepatotoxicity in clinical applications. Betulinic acid (BA) is a natural triterpenoid that protects against liver damage. However, the underlying mechanism has not yet been elucidated. The purpose of this study was to evaluate the ameliorative effects of BA on CYP-induced hepatotoxicity and further clarify the underlying mechanism. BA pretreatment mitigated CYP-induced liver oxidative damage by alleviating histopathological lesions, reducing reactive oxygen species (ROS) accumulation, and restoring the mRNA expression of antioxidant enzymes (Cu-Sod, Mn-Sod, Cat, and Gsh-Px). BA treatment also suppressed CYP-induced oxidative stress by activating the NRF2 pathway and inhibiting the MAPK signaling pathway. Moreover, BA attenuated CYP-triggered hepatic apoptosis by suppressing excessive mitochondrial fission, boosting mitochondrial fusion, and ameliorating pro-apoptotic protein expression (CASP9 and the ratio of BCL-2/BAX) by blocking the oxidative stress-activated mitochondrial apoptotic pathway. Furthermore, PD98059 (an inhibitor of ERK) and/or BA abated CYP-provoked hepatotoxicity by inhibiting the ERK-MAPK and mitochondrial apoptotic pathways, implying that deactivation of the ERK-mediated mitochondrial apoptotic pathway contributed to the hepatoprotective efficacy of BA against CYP-induced oxidative stress. Therefore, BA could be used as a complementary medicine in patients undergoing CYP treatment owing to its hepatoprotective effects.
摘要：
IN BRIEF: Postovulatory oocyte aging significantly compromises oocyte quality, resulting in fertilization failure, abnormal embryonic development, and unfavorable outcomes in assisted reproductive technology (ART). Epigallocatechin-3-gallate (EGCG), a polyphenolic compound found in green tea, demonstrates the ability to effectively reduce excessive oxidative stress in oocytes and improve the quality of aged oocytes both in vitro and in vivo, offering promising potential for enhancing the success of ART and efficiency in livestock breeding. ABSTRACT: Oxidative stress-mediated postovulatory aging (POA) significantly compromises oocyte quality and impairs subsequent embryonic developmental competence, thereby reducing the efficiency of assisted reproductive technologies (ART) and livestock breeding. As the most abundant polyphenolic compound in green tea, epigallocatechin-3-gallate (EGCG) has demonstrated notable antioxidant activity. However, the mechanisms by which EGCG modulates POA remain largely unclear. This study aimed to investigate whether EGCG delays POA both in vitro and in vivo by alleviating oxidative stress. During in vitro aging, metaphase II (MII) stage mouse oocytes were treated with various concentrations of EGCG for 12 h. EGCG treatment attenuated abnormal spindle formation and restored mitochondrial function. Furthermore, EGCG reduced reactive oxygen species levels and apoptosis, thereby mitigating oxidative damage associated with postovulatory aging. Notably, these improvements led to significantly enhanced embryonic developmental potential. In the in vivo experiments, mice received daily EGCG injections for 6 consecutive days. The results demonstrated that EGCG significantly improved oocyte quality during POA and alleviated adverse pregnancy outcomes. Taken together, our findings suggest that EGCG is a promising agent for preventing postovulatory oocyte aging and provides a basis for further strategies aimed at improving the success of ART and livestock breeding.
摘要：
Objective: Spirometra mansoni is a crucial zoonotic parasite. Its larvae are more harmful than adult worms due to their ability to migrate through the host's tissues and organs. Therefore, it is necessary to establish an animal model of spargana for observing pathological changes and exploring diagnostic techniques. Methods: In this study, we infected Kunming mice and cats without any pathogens by feeding sparganum (with the scolex and neck) in order to understand the infection cycle of S. mansoni and explore the preservation host of sparganosis. The infection of S. mansoni was determined by fecal detection and enzyme-linked immunosorbent assay (ELISA). Results: In the model of cats, the eggs of S. mansoni were found in the feces ten days after the infection. The serum-specific IgG antibodies against S. mansoni were positive in experimental groups (mice and cats), and after sixty days, the S. mansoni worms isolated from experimental groups were collected. Conclusion: In conclusion, the experimental results show that mice and cats can be stably infected with S. mansoni through feeding sparganum (with the scolex and neck). The infection method of this study has the potential to establish a practical model for investigating the diagnostic process of S. mansoni, laying the groundwork for application and development. ELISA was used to diagnose mice and cats infected with sparganosis mansoni, providing a case for non-invasive identification of animal sparganosis.
摘要：
Toxoplasma gondii is a widespread protozoan with a complex life cycle, characterized by transitions between various hosts and developmental stages, each tailored to a specific niche within its host. However, the regulatory mechanisms governing these life cycle transitions are not well understood. In this study, we investigated the AP2 factor AP2X-1, which is expressed during the tachyzoite and bradyzoite stages but decreases in the mature merozoite stage. Knockout of ap2X-1 significantly impaired tachyzoite invasion and replication while increasing the frequency of bradyzoite differentiation. As a component associated with the HDAC3/MORC complex, loss of ap2X-1 led to the upregulation of bradyzoite- and sexual stage-specific genes. Single-cell sequencing revealed that the ap2X-1 knockout strain exhibited a mixed population of tachyzoite-, bradyzoite-, merozoite-, and sporozoite-like parasites. Cleavage under targets and tagmentation analysis revealed a substantial overlap between AP2X-1 and the HDAC3/MORC complex at the promoters of bradyzoite- and sexual stage-specific genes. Additionally, assay for transposase-accessible chromatin with high-throughput sequencing analysis demonstrated that AP2X-1 influences chromatin compaction and accessibility, suggesting that AP2X-1 may modulate the function of the HDAC3/MORC complex to facilitate the repression of bradyzoite differentiation and sexual commitment. Loss of ap2X-1 resulted in significant attenuation of T. gondii virulence and decreased brain cyst formation in vivo. These findings identify AP2X-1 as a critical negative regulator of T. gondii sexual development.IMPORTANCEToxoplasma gondii undergoes a complex life cycle characterized by alternating developmental stages. The genetic reprogramming mechanisms driving these stage transitions remain largely unknown. In this study, we identified the AP2 factor AP2X-1 as a critical regulator important for T. gondii growth and life cycle progression. Our findings suggest that AP2X-1 functions as a repressor by modulating the function or influencing the association of the HDAC3/MORC complex at the promoters of bradyzoite- and sexual stage-specific genes, leading to chromatin compaction, restricting DNA accessibility and thereby repressing the transcription of genes required for bradyzoite formation and sexual commitment. Deletion of ap2X-1 significantly reduced T. gondii virulence and its ability to form brain cysts. These findings reveal a previously unknown regulatory pathway controlling sexual development in T. gondii, providing new insights into its underlying mechanisms.
通讯机构：
[Wang, ND ] H;Hunan Agr Univ, Hunan Prov Key Lab Prot Engn Anim Vaccines, Lab Funct Prote LFP, Changsha, Peoples R China.;Hunan Agr Univ, Coll Vet Med, Res Ctr Reverse Vaccinol RCRV, Changsha, Peoples R China.
关键词：
Cap;HnRNP K;Mitochondrial apoptosis;Porcine circovirus type 3
摘要：
Porcine circovirus type 3 (PCV3) is a globally emerging circovirus affecting pigs and other animals. The capsid protein (Cap) is the sole structural protein of PCV, with a crucial role in virus evolution and pathogenesis. Through interactions with host factors, Cap enables viral entry, transport, and replication while modifying various cellular processes. Cap protein-induced apoptosis has important implications for viral pathogenesis, but remains poorly defined. Herein, we demonstrated for the first time that PCV3 Cap induced cell cycle arrest of PK-15 cells in S-phase and initiated apoptosis via a mitochondrial Caspase-9-dependent pathway. Truncation analysis localized the apoptotic determinant to the N-terminal 1-34 aa of PCV3 Cap and heterogeneous nuclear ribonucleoprotein K (hnRNP K) was identified as a host protein that binds to PCV3 Cap. Overexpression of hnRNP K reduced PCV3 Cap-induced release of Cyt-c into the cytoplasm, implying a regulatory role in apoptosis. Based on structural modelling and molecular docking, amino acids at sites 24 and 27 of Cap from PCV3 variants, which define genotypes (PCV3a/b/c), affected binding with hnRNP K. Specifically, PCV3c Cap (V24/K27 and V24/R27) had higher affinity than PCV3a Cap (A24/R27) or PCV3b Cap (A24/K27), consistent with its superior apoptosis-inducing capacity compared to PCV3a/b variants, highlighting the importance of Cap interactions with hnRNP K. In summary, we identified novel molecular determinants of PCV3 pathogenesis that will inform development of vaccines and diagnostics.
Porcine circovirus type 3 (PCV3) is a globally emerging circovirus affecting pigs and other animals. The capsid protein (Cap) is the sole structural protein of PCV, with a crucial role in virus evolution and pathogenesis. Through interactions with host factors, Cap enables viral entry, transport, and replication while modifying various cellular processes. Cap protein-induced apoptosis has important implications for viral pathogenesis, but remains poorly defined. Herein, we demonstrated for the first time that PCV3 Cap induced cell cycle arrest of PK-15 cells in S-phase and initiated apoptosis via a mitochondrial Caspase-9-dependent pathway. Truncation analysis localized the apoptotic determinant to the N-terminal 1-34 aa of PCV3 Cap and heterogeneous nuclear ribonucleoprotein K (hnRNP K) was identified as a host protein that binds to PCV3 Cap. Overexpression of hnRNP K reduced PCV3 Cap-induced release of Cyt-c into the cytoplasm, implying a regulatory role in apoptosis. Based on structural modelling and molecular docking, amino acids at sites 24 and 27 of Cap from PCV3 variants, which define genotypes (PCV3a/b/c), affected binding with hnRNP K. Specifically, PCV3c Cap (V24/K27 and V24/R27) had higher affinity than PCV3a Cap (A24/R27) or PCV3b Cap (A24/K27), consistent with its superior apoptosis-inducing capacity compared to PCV3a/b variants, highlighting the importance of Cap interactions with hnRNP K. In summary, we identified novel molecular determinants of PCV3 pathogenesis that will inform development of vaccines and diagnostics.
摘要：
Avian gout (AG) is detrimental to the survival and production performance of poultry and effective drugs are lacking. Caulis sinomenii has shown clinical efficacy against arthritis and may have potential value in AG prevention and treatment. In the present study, the components and targets of C. sinomenii and AG-related targets were identified using relevant databases. The common targets, target interactions, and signaling pathways involved in the prevention and treatment of AG by C. sinomenii were determined using software to explore the potential mechanisms of action. Sixteen components of C. sinomenii, eight of which were active ingredients with 351 targets and 2993 AG-related targets, were identified using several databases. A total of 156 common targets were associated with 202 biological processes and 34 pathways. Toll-like receptor 4 (TLR4) and prostaglandin endoperoxide synthase 2 were core targets. These targets may exert therapeutic effects on AG through four pathways: the nucleotide-binding oligomerization domain (NOD)-like receptor, mammalian target of rapamycin, TLR, and mitogen-activated protein kinase signaling pathways. In summary, C. sinomenii has potential therapeutic efficacy against AG through multicomponent, multi-target, and multi-pathway mechanisms.
摘要：
Citrinin (CTN) is commonly found in animal feed and stored grains and poses a serious threat to human and animal health. Formation of the IP3R1-GRP75-VDAC1 complex has been shown to play a key role in intestinal defense against harmful stimuli, but the mechanism of its action in CTN-exposure-induced enterotoxicity is not clear. Therefore, the aim of this study was to investigate the role of the IP3R1-GRP75-VDAC1 complex in CTN-exposure-induced intestinal and IPEC-J2 monolayer cell damage in mice. It was shown that CTN exposure triggered intestinal cell pyroptosis and increased IP3R1-GRP75-VDAC1 complex formation as well as mitochondrial levels of calcium ions and mitochondrial reactive oxygen species (mtROS). And mtROS is considered to be a key factor in cellular pyroptosis. Therefore, the removal of mtROS by using Mito-Tempo was found to attenuate CTN-exposure-induced cellular pyroptosis but failed to attenuate mitochondrial calcium ion overload. However, silencing of GRP75 alleviated CTN-exposure-induced increases in the level of mtROS, mitochondrial calcium ions, and subsequent cellular pyroptosis. Therefore, this study confirms that CTN exposure induces cellular juxtaposition in intestinal tissues and points out that mitochondrial oxidative stress mediated by the IP3R1-GRP75-VDAC1 complex is a key mechanism by which CTN exposure triggers intestinal cellular pyroptosis.
摘要：
The T-2 toxin, originating from a Fusarium species, is a mycotoxin that can adversely affect animal health. Melatonin (MT) is a natural hormone recognized for its properties that reduce inflammation and act as an antioxidant. However, MT's capacity to alleviate intestinal harm from T-2 toxin remains incompletely explored. Employing postweaning piglets, this research investigates MT's prophylactic impact on T-2 toxin-induced enterotoxicity. The results indicate that MT improved growth performance in piglets exposed to T-2 toxins while also enhancing intestinal barrier function. Such effects probably stem from MT's ability to reduce colonic oxidative stress and inflammation. Further findings suggest that these changes are closely associated with MT-induced remodeling of intestinal microbiota and an increase in short-chain fatty acid (SCFA) levels in the intestine. MT therefore alleviates T-2 toxin intestinal damage; gut microbiota are the key to this process.
