Abstract Background The Three-River-Source Region of the Qinghai-Tibet Plateau is a hyperendemic focus for echinococcosis, with Echinococcus granulosus, E. multilocularis, and E. shiquicus circulating between definitive canid hosts (dogs and foxes) and intermediate hosts (livestock and rodents). However, the extent of environmental contamination by Echinococcus eggs remains understudied and poses significant risks to human and animal health. Methods From 2019 to 2021, we collected 631 canid fecal samples (296 from dogs and 335 from foxes) and 398 adjacent soil samples across endemic counties in the Three-River-Source Region of the Qinghai-Tibet Plateau. Multiplex real-time PCR was employed to detect Echinococcus species DNA in feces and soil samples. Results The overall Echinococcus prevalence in canid feces was 7.13% (45/631), with 3.01% in E. multilocularis (19/631), 2.06% in E. granulosus (13/631), and 2.06% in E. shiquicus (13/631). Foxes presented increased E. multilocularis (3.88%, 13/335) and E. shiquicus (2.69%, 9/335) infections, whereas dogs presented increased E. granulosus prevalence (2.70%, 8/296). Soil contamination with Echinococcus species was detected in 2.51% (10/398) of the samples. The primary contaminants were E. multilocularis and E. shiquicus (1.01% each, 4/398), whereas E. granulosus was less frequent (0.50%, 2/398). Moreover, the soil near fox feces was contaminated with both E. multilocularis and E. shiquicus, whereas the dog-associated soil was contaminated with all three species. Conclusions This study suggests widespread environmental deposition of Echinococcus eggs on the Qinghai-Tibet Plateau, driven by canid defecation. If these eggs remain viable, their persistence in soil would indicate a potential zoonotic transmission risk, highlighting the need for integrated control strategies targeting both domestic and wild canids. Graphical abstract

Abstract Zika virus (ZIKV), a mosquito-borne flavivirus, has emerged as a public health concern. Although asymptomatic or mild febrile illness in adults, ZIKV infection can lead to severe outcomes in pregnant women, including fetal microcephaly and congenital Zika syndrome. A hallmark of ZIKV pathogenesis is the virus-induced remodeling of the endoplasmic reticulum into specialized replication organelles (ROs), which function as spatially organized hubs that actively couple viral RNA synthesis with the host translation machinery. These specialized ROs enhance viral genome amplification, prioritizes viral mRNA translation, and minimizes the cytosolic exposure of immunostimulatory viral double-stranded RNA intermediates. This review explores how this spatial coupling underlies the unique tropism of ZIKV towards neural progenitor cells, a characteristic that sets it apart from other neurotropic flaviviruses. In particular, ZIKV adaptations that simultaneously promote viral replication and disrupt host neurogenesis are critically discussed.

Colistin resistance genes have attracted increasing global attention due to their plasmid-mediated transmissibility. To clarify the transmission dynamics of colistin resistance genes in the endangered crested ibis (Nipponia nippon) and its environment, surveillance for mcr-1 was conducted at a captive breeding center in Deqing, Zhejiang, China, from 2017 to 2024. Genomic sequencing was used to analyze the phylogenetic relationships among the Enterobacteriaceae isolates harboring these resistance genes. A total of 2660 fecal samples of crested ibises were collected seasonally. There was only one episode of mcr-1 occurrence with a total of 144 mcr-1-positive strains in crested ibises. The prevalence was first detected at 8% in April 2021, surged to 25% in July, and then declined to 10.6% in October. Additionally, mcr-1-harboring Escherichia coli isolates were also found in feces from wild egrets (33%), the staff member's dog (8.3%), and effluent sewage, but not in the staff member, loach samples, or water sources. Phylogenetic analysis showed that the egret-derived isolate BLM86 and the crested ibis-derived isolate ZHM12 belonged to the same ST58-cgST28300 type; no core-genome SNP differences were detected. All 36-deeper analyzed mcr-1-positive isolates displayed multidrug resistance, with four distinct mcr-1-containing plasmid types identified: IncI2 (20/36, 55.6%), IncX4 (19.4%), IncP1 (11.1%), and IncHI2 (13.9%). Phylogenetic analysis indicated that several of these crested ibis-derived isolates were closely related to those from egrets, implying potential environmental transmission routes to both the crested ibis and egrets. Mcr-1 was no longer detected in the crested ibis population after implementation of biosecurity interventions. Our findings demonstrate that wild egrets could act as potential vectors for the environmental transmission of mcr-1 to captive crested ibises, highlighting the importance of interspecies barriers in antimicrobial resistance (AMR) containment.

Alveolar echinococcosis (AE) is a severe and life-threatening parasitic disease caused by Echinococcus multilocularis. Liver fibrosis is a significant pathological feature of advanced AE, characterized by the excessive production and accumulation of extracellular matrix (ECM). However, the precise underlying mechanism remains largely unknown. In this study, we show that the long noncoding RNA gm40262, predominantly expressed in hepatic stellate cells (HSCs), is upregulated in AE. Interestingly, its knockdown leads to liver fibrosis resolution, accompanied by a substantial suppression of parasite growth. Gm40262 functions by targeting miR-193b-5p to activate HSCs and stimulate their proliferation in a TGF-β-dependent manner, thereby promoting ECM production by upregulating Col1α1. Moreover, gm40262 is also involved in inflammation through the gm40262-miR-193b-5p-TLR4 axis. Our findings suggest that gm40262 plays a pivotal role in parasite-induced liver fibrosis through multiple mechanisms, highlighting its potential as a therapeutic target for hepatic fibrosis. IMPORTANCE:Echinococcus multilocularis is a tiny parasite with significant medical implications. The chronic parasitism of E. multilocularis in the liver generally leads to liver fibrosis, but the underlying mechanisms are poorly understood. We herein show that gm40262, a long noncoding RNA predominantly expressed in hepatic stellate cells (HSCs), is involved in hepatic fibrogenesis during infection by activating HSCs and promoting extracellular matrix production. The gm40262-orchestrating fibrogenesis occurs through the gm40262-miR-193b-5p-TLR4 and gm40262-miR-193b-5p-Col1α1 axes. The knockdown of gm40262 remarkably alleviates liver fibrosis, with decreased parasite growth. Our findings reveal a key role of gm40262 in liver fibrosis during E. multilocularis infection, rendering it a therapeutic target for hepatic fibrosis.

Echinococcosis, a serious zoonotic parasitic disease caused by tapeworms of the genus Echinococcus, is clinically characterized by a long latent period of up to 10 years. An accurate diagnosis is critical for the efficient management and treatment of patients. The aim of this study was to identify robust diagnostic signatures for echinococcosis. Using co-immunoprecipitation and RNA sequencing (RNA-seq), we comparatively profiled the Argonaute 2-binding microRNAs (abmiRNAs) in hepatocytes of Echinococcus multilocularis-infected mice. Using receiver operating characteristic curve (ROC) analysis, we established quantitative PCR (qPCR) assays based on circulating liver-specific abmiRNAs and their combinations, and further validated these assays using blinded serum samples from infected mice and patients. Three abmiRNAs were identified as being predominantly expressed in the liver: miR-192-5p, miR-122-5p and miR-21a-5p. While these three abmiRNAs are upregulated in liver cancer and hepatitis virus infections, we found that all circulating abmiRNAs were significantly and gradually downregulated as the Emultilocularis infection progressed; however, they were rapidly upregulated following anthelmintic treatment. The qPCR assays targeting these circulating abmiRNAs and their combinations showed high sensitivity and specificity. Individual circulating abmiRNAs and their combinations accurately distinguished infections in both mice (n = 50) and humans (n = 117), with the combination of miR-192-5p and miR-122-5p being particularly effective in distinguishing infections. This combination was also sensitive to anthelmintic treatment. These results suggest that circulating miR-192-5p and miR-122-5p are serum signatures for the diagnosis and prognostic management of echinococcosis.

Parasite diseases pose a significant threat due to the serious harm they cause to hosts, and developing affordable drugs for treatment presents enormous challenges. "Target repurposing" has emerged as a promising strategy to overcome this difficulty by leveraging the drug targets for human diseases and matching them with homologs in parasites. The PI3K/AKT signaling pathway, known for its crucial role in regulating malignant cell proliferation, migration, and invasion, has been validated as a drug target for cancer. Interestingly, the PI3K/AKT signaling pathway is also involved in parasite development, infection, and survival within the hosts. During infection, parasites can exploit the host's PI3K/AKT signaling, which influences immune cell apoptosis, macrophage activation, regulatory T cell functions, and the communication between immune cells, thus facilitating chronic infection. The immunoregulatory functions of the PI3K/AKT signaling make its inhibitors promising candidates for the control of parasite diseases. This review mainly outlines the characteristics and functions of the PI3K/AKT signaling pathway during parasite infection and further provides an overview of the drugs targeting the PI3K/AKT signaling pathway for treating parasite diseases.

Citation: Li R, Zheng Y and Cho WC (2024) Editorial: Insights in RNA: 2022. Front. Genet. 15:1382435. doi: 10.3389/fgene.2024.1382435

Eimeria spp.can invade the different intestines of chickens.Among them, Eimeria acervulina (E.acervulina, Ea) is the most virulent, and it is characterized by disruption of the intestinal nutrient uptake mechanism

本实验室前期构建的基于毒力基因改造的减毒单增李斯特菌(LM-1)被证实是一种较为安全的疫苗载体.为构建表达鸡柔嫩艾美耳球虫(E.tenella)毒力蛋白MIC3的重组减毒单增李斯特菌(LM)疫苗候选株并评估其对雏鸡的免疫保护效果,本研究在LM-1基础上采用同源重组的方法将MIC3蛋白关键功能区(aa159～aa252)编码基因(475 bp～756 bp)定点整合至LM-1基因组中,并与LM溶血素O(LLO,其编码基因为Rly)融合表达获得重组LM-1-MIC3菌株并经PCR鉴定;western blot检测LM-1-MIC3中MIC3和LLO的表达;测定细菌体外生长曲线检测LM-1-MIC3的体外生长能力;将野生菌株EGO-e、LM-1、LM-1-MIC3分别感染雏鸡72 h和144 h后,分别剖杀,取各组雏鸡肝脏和脾脏,经菌落计数对雏鸡肝脾载菌量进行测定并评估LM-1-MIC3在雏鸡体内的增殖能力;将野生株EGD-e、LM-1和LM-1-MIC3感染雏鸡并结合临床症状及致死率评估LM-1-MIC3对雏鸡的安全性;将LM-1-MIC3免疫雏鸡并建立E.tenella感染模型,研究其对雏鸡抗球虫感染的免疫保护效果.PCR鉴定结果显示正确构建重组菌株LM-1-MIC3;western blot结果显示LM-1-MIC3分泌的融合蛋白LLO-MIC3能够在培养上清液中大量表达;细菌体外培养结果显示LM-1-MIC3体外生长能力与EGD-e和LM-1无显著差异,表明融合LLO的MIC3不会影响LM-1的生长;雏鸡肝脾载菌量测定结果显示,与EGD-e组相比,各时间段LM-1-MIC3组雏鸡肝脾载菌量均极显著降低(P＜0.01),感染LM-1-MIC3后雏鸡均存活且无明显临床症状,表明LM-1-MIC3对雏鸡致病力高度减弱,可以作为减毒疫苗载体;免疫保护试验结果显示在攻虫10 d后,LM-1-MIC3免疫组雏鸡的存活率高于LM-1免疫组及PBS对照组,且该组鸡无明显临床症状,剖杀后盲肠病变评分极显著低于LM-1和PBS组(P＜0.001),表明LM-1-MIC3对雏鸡有良好的免疫保护效果.本研究构建了重组减毒株LM-1-MIC3,并证实该重组菌对雏鸡具有较好的免疫保护效果,为减毒LM载体疫苗在防控重要动物传染病中的研究与应用奠定了科学基础.

Through prokaryotic expression,antibody preparation and tissue distribution of RNA binding protein EWS(EmEWS)gene of Echinococcus multilocularis(E.multilocularis),the gene characteristics and binding ability of EmEWS with RNA were preliminarily explored.A pair of primers were designed ac-cording to the sequence from WormBase.EmEWS gene was amplified by RT-PCR and a recombinant ex-pression plasmid was constructed.The recombinant EmEWS protein was induced expressed and purified.EmEWS polyclonal antibody was prepared and identified by Western blot,real-time fluorescence quantita-tive PCR,immunofluorescence assay and dual fluorescence assay.The results showed that the full length of EmEWS gene was 1 485 bp,encoding 495 amino acids.The EmEWS gene was highly expressed in the pri-mary cells of E.multilocularis and lowest in the protoscoleces.Bioinformatics analysis showed that EWS of Echinococcus granulosus was 98.3％similar to EmEWS,and the RBD domain of EmEWS was highly con-served in tapeworms.The size of recombinant EmEWS protein was about 54 ku,and the prepared poly-clonal antibody could specifically recognize the native EmEWS protein.Immunoflurescence showed that EmEWS mainly distributed on the cell membrane.Double fluorescence assay demonstrated that EmEWS could bind to UG-rich RNA sequences.Taken together,it is speculated that EmEWS may be involved in the formation of small RNA as a transcription regulator and contribute to nuclear protein transport.

Heat shock protein 60 (HSP60) is an unique member of the heat shock protein family, being involved in parasite infections. To cope with harsh environments where parasites live, HSP60s are indispensable and involved in a variety of biological processes. HSP60s have relative low similarity among parasites, but their ATPase /Mg2+ active sites are highly conserved. The interactions of HSP60s with signaling pathway regulators in immune cells suggest a crucial role in immune responses, rendering them a potential therapeutic target. This paper reviews the current understandings of HSP60s in parasitic helminths in aspects of molecular characteristics, immunoregulatory responses and HSP60-based therapeutics.

Cyst echinococcosis, caused by Echinococcus granulosus, remains a zoonotic disease posing a great threat to public health and meat production industry. Sheep infected with E. granulosus show relatively high abundance of egr-miR-71 in the sera, but its role is unknown. Using bioinformatics and cell migration and Transwell assays, we comparatively analyzed the proteomes and cell invasion of sheep PBMCs in response to egr-miR-71 overexpression. The results showed that the egr-miR-71 induced a total of 157 proteins being differentially expressed and mainly involved in immune responses. In sheep PBMCs, egr-miRNA-71 overexpression induced significant downregulation of macrophage migration inhibitory factor (MIF) and accordingly promoted cell migration and invasion compared with the control. The results will provide a clue for further investigation of a role of circulating egr-miR-71 in immune responses during E. granulosus infection.

Chronic parasite infections in the liver pose a global threat to human and animal health, often occurring with liver fibrosis that leads to cirrhosis, liver failure, and even cancer. Hepatic fibrogenesis is a complex yet reversible process of tissue repair and is associated with various factors, including immune cells, microenvironment, gut microbiome, and interactions of the different liver cells. As a profibrogenic or antifibrogenic driver, microRNAs (miRNAs) are closely involved in parasite-induced hepatic fibrosis. This article updates the current understanding of the roles of miRNAs in hepatic fibrogenesis by parasite infections and discusses the strategies using miRNAs as candidates for diagnostics and therapeutics.

Tapeworms of the genus Dibothriocephalus are widely distributed throughout the world, some of which are agents of human diphyllobothriasis, one of the most important fish-borne zoonoses caused by a cestode parasite. Genomic and transcriptomic data can be used to develop future diagnostic tools and epidemiological studies. The present work focuses on a comparative analysis of the transcriptomes of adult and plerocercoid D. dendriticus and the identification of their differentially expressed genes (DEGs). Transcriptome assembly and analysis yielded and annotated 35,129 unigenes, noting that 16,568 (47%) unigenes were not annotated in known databases, which may indicate a unique set of expressed transcripts for D. dendriticus. A total of 8022 differentially expressed transcripts were identified, including 3225 upregulated and 4797 downregulated differentially expressed transcripts from the plerocercoid and adult animals. The analysis of DEGs has shown that among the most differentially expressed genes, there are important genes characteristic of each stage. Thus, several genes are characteristic of D. dendriticus plerocercoids, including fatty acid-binding protein and ferritin. Among the most highly expressed DEGs of the adult stage of D. dendriticus is the Kunitz-type serine protease inhibitor, in two putative isoforms. The analyses of GO and KEGG metabolic pathways revealed that a large number of the DEGs of D. dendriticus are associated with the biosynthesis of various substances such as arginine and folate, as well as with various metabolic pathways such as galactose metabolism, selenocompound metabolism, and phosphonate and phosphinate metabolism. This will contribute to further research aimed at identifying targets for new generation drugs and the development of specific vaccines.

Liver fibrosis is one of the histopathological characters during Echinococcus multilocularis infection. The activation of hepatic stellate cells (HSCs) is a key event in the development of liver fibrosis. However, the molecular mechanism of HSC activation in the E. multilocularis infection-induced liver fibrosis remains largely unclear. Here, we reported that mmu-miR-342-3p was most dominantly expressed in HSCs and was upregulated in the HSCs in response to E. multilocularis infection. We further showed that mmu-miR-342-3p was able to bind to the 3' UTR of the Zbtb7a gene and regulated its expression. Moreover, mmu-miR-342-3p expression was negatively correlated with its target gene Zbtb7a in HSCs during E. multilocularis infection. Knockdown of mmu-miR-342-3p promoted the expression of Gfap in the activated HSCs in vitro. In the E. multilocularis-infected mice, knockdown of mmu-miR-342-3p suppressed the expression of α-Sma, Col1α1, and TGF-β but promoted the expression of Gfap. Therefore, mmu-miR-342-3p is a key regulator for activation of HSCs, and inhibiting mmu-miR-342-3p to suppressed Zbtb7a-mediated TGF-β signaling in activated HSCs could be a novel strategy to treat liver fibrosis induced by E. multilocularis.

Cystic echinococcosis (CE) is a global zoonotic disease caused by Echinococcus granulosus, posing a great threat to human and animal health. MiRNAs are small regulatory noncoding RNA involved in the pathogenesis of parasitic diseases, possibly via exosomes. Egr-miR-71 has been identified as one of the miRNAs in the blood of CE patients, but its secretory characteristics and functions remains unclear. Herein, we studied the secretory and biological activity of exosomal egr-miR-71 and its immunoregulatory functions in sheep peripheral blood mononuclear cells (PBMCs). Our results showed that egr-miR-71 was enriched in the exosome secreted by protoscoleces with biological activity. These egr-miR-71-containing exosomes were easily internalized and then induced the dysregulation of cytokines (IL-10 and TNF-α), nitric oxide (NO) and key components (CD14 and IRF5) in the LPS/TLR4 pathway in the coincubated sheep PBMCs. Similarly, egr-miR-71 overexpression also altered the immune functions but exhibited obvious differences in regulation of the cytokines and key components, preferably inhibiting proinflammatory cytokines (IL-1α, IL-1β and TNF-α). These results demonstrate that exosomal egr-miR-71 is bioactive and capacity of immunomodulation of PBMCs, potentially being involved in immune responses during E. granulosus infection.

[目的]掌握浙江省德清县驯养朱鹮大肠杆菌的耐药性、耐药基因与分子特征,为治疗朱鹮大肠杆菌感染提供基础数据.[方法]采集该地驯养朱鹮新鲜粪便样本,采用分离培养、形态观察、生化鉴定及16S rDNA序列分析鉴定大肠杆菌;进而采用肉汤稀释法检测环丙沙星、庆大霉素、丁胺卡那等7种抗菌药对源自不同朱鹮的大肠杆菌分离株菌株的最小抑菌浓度(minimal inhibitory concentration,MIC);选择代表性菌株,通过PCR和测序鉴定其携带qnrS1、gyrA、gyrB等9种喹诺酮类耐药基因的情况,并分析其蛋白关键氨基酸位点突变与耐药性的关系,采用接合转移分析耐药质粒水平转移情况及其与耐药性的关系,应用卡方检验(Chi-square test)和费歇尔精确检验(Fisher exact test)分析分离株耐药表型与朱鹮年龄、耐药基因的相关性.[结果]本研究共采集了 98只朱鹮的粪便,经分离鉴定均获得了大肠杆菌;源自不同朱鹮的98株大肠杆菌对喹诺酮类环丙沙星呈现高度耐药(耐药率为65.3％,64/98),对其余6种药物高度敏感(敏感率均＞90％),朱鹮年龄与环丙沙星耐药性极显著相关(P＜0.01);选取的31株分离株喹诺酮类耐药基因marR、gyrA、parC和gyrB的突变株占比在3.2％～80.6％之间,携带质粒耐药基因qnrS1的菌株占比22.6％(7/31);耐药基因阳性菌株占比93.5％(29/31)并分布于6个耐药基因型,其中,marR/gyrA/parC10 株、marR/qnrS1 6 株、marR 6 株、gyrA/parC3 株、marR/gyrA 3 株、marR/gyrB/qnrS11株;环丙沙星耐药表型与耐药基因显著相关(P=0.026),gyrA和parC单基因或联合突变均与喹诺酮耐药显著相关(P=0.038);大肠杆菌分离株qnrS1基因一次接合成功率为28.6％(2/7),并可导致受体菌对喹诺酮类药物的耐药性增加.[结论]本研究鉴定了朱鹮群体大肠杆菌耐药性,并明确了朱鹮源大肠杆菌对喹诺酮类耐药的主要原因是gyrA和parC单基因或联合突变并携带可水平转移的耐药基因qnrS1,本研究结果为人工驯养朱鹮救护和耐药性监测提供参考依据,提示应重视朱鹮大肠杆菌对喹诺酮类药物的耐药性.

目的 通过扩增多房棘球绦虫(Echinococcus multilocularis)RNA结合蛋白28(RBP28)基因并构建重组表达质粒,诱导表达并纯化重组RBP28蛋白,探索多房棘球绦虫RBP28的分子特征.方法 根据WormBase数据库RBP28基因序列设计特异性引物;RT-PCR扩增RBP28基因并构建重组表达质粒,诱导表达并纯化重组RBP28蛋白,免疫新西兰大白兔,制备多克隆抗体;用Western blot检测多克隆抗体特异性,并通过免疫荧光观察RBP28在多房棘球蚴及其细胞中的分布;利用免疫共沉淀技术,获得与RBP28互作的蛋白沉淀产物后,对二者进行蛋白银染和质谱鉴定.结果 RBP28编码基因大小约为1 245 bp;重组RBP28相对分子质量约为Mr 50 000;Western blot结果表明,多克隆抗体能够特异性识别重组蛋白RBP28及多房棘球蚴全蛋白中的天然RBP28蛋白,但与猪带绦虫、泡状带绦虫和细粒棘球绦虫无明显的抗原交叉反应;免疫荧光定位表明,RBP28主要分布在多房棘球蚴的生发层,在多房棘球蚴细胞则主要分布于细胞质中;对银染和质谱鉴定结果比较分析,获得了 7种可能与RBP28相互作用的蛋白质.结论 本研究确定了 RBP28蛋白编码基因的大小、分子量及其多克隆抗体的特异性.通过免疫荧光定位,发现RBP28主要分布在多房棘球蚴的生发层,并利用质谱鉴定获得了 7种与RBP28有互相作用的蛋白质,为研究多房棘球绦虫病的致病机制奠定了基础.

目的 建立EdU(5-ethynyl-2,-deoxyuridine)标记多房棘球蚴体内成体干细胞(neoblasts)的方法,用于观察其细胞形态和分布特点.方法 采用0.25％胰蛋白酶消化法从多房棘球蚴包囊中分离原代细胞,分别对原代细胞和多房棘球蚴进行EdU染色,利用激光共聚焦显微镜检测EdU阳性细胞.结果 用建立的EdU标记方法检查多房棘球蚴体内成体干细胞,EdU阳性细胞体积较小,细胞核较大,呈球形或近球形.在成熟的原头节头节中,EdU阳性细胞主要分布在其中后端,呈分散分布,且原头节头节是否外翻在EdU阳性细胞数量上无明显不同.同时,大多数EdU阳性细胞也表达磷酸化组蛋白H3.结论 建立的EdU标记多房棘球蚴成体干细胞的方法可靠,简单,易操作,为该细胞的功能研究提供了新的技术手段.