MicroRNA-187 inhibits pentylenetetrazol-induced neuronal apoptosis and alleviates development of epilepsy in epileptic rats by regulating SPRY1 expression

Purpose: To explore the role of microRNA-187 on the pathological process of epilepsy. Methods: The seizure score of epileptic rats was evaluated according to Racine’s scale. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to determine the expression levels of microRNA-187 (miR-187). Western blot technique was conducted to assess the expression levels of caspase 3, B-cell lymphoma-2 (BCL-2), and poly (ADP-ribose) polymerase (PARP)] and activation of phosphatase and tensin homolog (PTEN)/PI3K/AKT cascade. Caspase 3 colorimetric assay kit was employed to evaluate the activity of caspase 3. Dual-luciferase reporter gene system was used to explore the regulating mechanisms of miR-187 and protein sprouty homolog 1 gene (SPRY1). Results: The results showed that miR-187 was aberrantly downregulated in the hippocampus regions of pentylenetetrazol (PTZ)-treated rats compared to normal rats (p < 0.05). Furthermore, PTZ promoted caspase 3-dependent neuronal apoptosis by increasing the expression of pro-apoptosis protein PARP and decreasing the expression levels of BCL-2 in rats. On the other hand, overexpression of miR-187 downregulated SPRY1 as well as PTEN (p < 0.05), thereby activating the downstream PI3K/AKT signaling pathway. Notably, the effects of upregulated miR-187 on neuronal apoptosis and epilepsy development in PTZ-induced rats was reversed by the concomitant overexpression of SPRY1 (p < 0.05). Conclusion: The results of this research show that overexpressed miR-187 alleviates the development of PTZ-induced neuronal apoptosis and epilepsy in epileptic rat models by regulating SPRY1 expression. These findings can hopefully be beneficial for the discovery of new therapeutic strategies for epilepsy treatment

A Dopa Decarboxylase Modulating the Immune Response of Scallop Chlamys farreri

BACKGROUND: Dopa decarboxylase (DDC) is a pyridoxal 5-phosphate (PLP)-dependent enzyme that catalyzes the decarboxylation of L-Dopa to dopamine, and involved in complex neuroendocrine-immune regulatory network. The function for DDC in the immunomodulation remains unclear in invertebrate. METHODOLOGY: The full-length cDNA encoding DDC (designated CfDDC) was cloned from mollusc scallop Chlamys farreri. It contained an open reading frame encoding a polypeptide of 560 amino acids. The CfDDC mRNA transcripts could be detected in all the tested tissues, including the immune tissues haemocytes and hepatopancreas. After scallops were treated with LPS stimulation, the mRNA expression level of CfDDC in haemocytes increased significantly (5.5-fold, P<0.05) at 3 h and reached the peak at 12 h (9.8-fold, P<0.05), and then recovered to the baseline level. The recombinant protein of CfDDC (rCfDDC) was expressed in Escherichia coli BL21 (DE3)-Transetta, and 1 mg rCfDDC could catalyze the production of 1.651±0.22 ng dopamine within 1 h in vitro. When the haemocytes were incubated with rCfDDC-coated agarose beads, the haemocyte encapsulation to the beads was increased significantly from 70% at 6 h to 93% at 24 h in vitro in comparison with that in the control (23% at 6 h to 25% at 24 h), and the increased haemocyte encapsulation was repressed by the addition of rCfDDC antibody (which is acquired via immunization 6-week old rats with rCfDDC). After the injection of DDC inhibitor methyldopa, the ROS level in haemocytes of scallops was decreased significantly to 0.41-fold (P<0.05) of blank group at 12 h and 0.47-fold (P<0.05) at 24 h, respectively. CONCLUSIONS: These results collectively suggested that CfDDC, as a homologue of DDC in scallop, modulated the immune responses such as haemocytes encapsulation as well as the ROS level through its catalytic activity, functioning as an indispensable immunomodulating enzyme in the neuroendocrine-immune regulatory network of mollusc

Novi VP2/VP3 rekombinantni senekavirus A izoliran u sjevernoj Kini

Senecavirus A (SVA), previously called the Seneca Valley virus, is the only member of the genus Senecavirus within the family Picornaviridae. This virus was discovered as a serendipitous finding in 2002 and named Seneca Valley virus 001 (SVV-001). SVA is an emerging pathogen that can cause vesicular lesions and epidemic transient neonatal a sharp decline in swine. In this study, an SVA strain was isolated from a pig herd in Shandong Province in China and identified as SVA-CH-SDFX-2022. The full-length genome was 7282 nucleotides (nt) in length and contained a single open reading frame (ORF), excluding the poly (A) tails of the SVA isolates. Phylogenetic analysis showed that the isolate shares its genomic organization, resembling and sharing high nucleotide identities of 90.5% to 99.6%, with other previously reported SVA isolates. The strain was proved by in vitro characterization and the results demonstrate that the virus has robust growth ability in vitro. The recombination event of the SVA-CH-SDFX-2022 isolate was found and occurred between nts 1836 and 2710, which included the region of the VP2 (partial), and VP3 (partial) genes. It shows the importance of faster vaccine development and a better understanding of virus infection and spread because of increased infection rates and huge economic losses. This novel incursion has substantial implications for the regional control of vesicular transboundary diseases, and will be available for further study of the epidemiology of porcine SVA. Our findings provide useful data for studying SVA in pigs.Senekavirus A (SVA), prije nazivan virusom doline Seneca Valley, jedini je pripadnik roda senekavirusa u porodici Picornaviridae. Virus je slučajno otkriven 2002. i nazvan virusom doline Seneca 001 (SVV-001). SVA je novi patogen koji može uzrokovati vezikularne lezije i prolaznu epidemiju novorođene prasadi s naglim gubicima u proizvodnji. U ovom je istraživanju soj SVA izoliran u populaciji svinja iz provincije Shandong u Kini i identificiran kao SVA-CHSDFX-2022. Kompletni genom izolata SVA imao je 7282 nukleotida (nt) u dužini i sadržavao je jedan otvoreni okvir za očitavanje (ORF), bez poli-A repova. Filogenetska je analiza pokazala da izolat u velikoj mjeri sadržava genomsku organizaciju i nukleotidne identitete, od 90,5 % do 99,6 %, s drugim poznatim SVA izolatima. Karakterizacija virusa je pokazala da ima veliku sposobnost rasta in vitro. Pronađena je rekombinacija izolata SVA-CH-SDFX-između nukleotida 1836 i 2710 što je uključilo regiju gena VP2 (parcijalno) i gena VP3 (parcijalno). Zbog visoke stope infektivnosti i golemih ekonomskih gubitaka važan je brži razvoj cjepiva i bolje razumijevanje zaraze. Rezultati ovog istraživanja pružaju korisne podatke za proučavanje SVA virusa, posebno s obzirom na njegovu epidemiologiju u svinja i regionalnu prekograničnu kontrolu vezikularnih bolesti

Analiza genskih varijacija rekombinantnog soja dobivenog iz triju linija virusa-2 reproduktivnog i respiratornog sindroma svinja

Since the rise of the porcine reproductive and respiratory syndrome virus (PRRSV) in China, gene mutations have frequently occurred. To understand the current prevalence and evolution of PRRSV in Shandong Province, 1,528 samples suspected of PRRSV were collected from local pig farms of different sizes. The complete genome sequence of the PRRSV strain SDLY-27 was determined by next-generation sequencing (NGS) technology. The genomic sequence of SDLY-27 was 15,363 nucleotides (nt) in length, comparative analysis of the whole genome sequence suggested that the homology between SDLY 27 and 81 PRRSV strains from China and other countries in genbank was 61.9 ~ 96.4%. This study is the first to detect recombinants from multiple recombination events among the Lineage 8 (JXA1-like strains), Lineage 5 (RespPRRSV-MLV and VR2332 strains) and Sublineage 1.5 (NADC34-like strains) in Shandong, China, and provides new data for the epidemiological study of PRRSV in China. This study enriches the epidemiological data on PRRSV in Shandong Province, China. It provides an important reference for the development of new vaccines and for the prevention and control of PRRSV in China.Usporedno sa širenjem virusa reproduktivnog i respiratornog sindroma svinja (PRRSV) u Kini, sve su češće bile i njegove genske mutacije. Kako bi se ustanovila trenutačna prevalencija i evolucija PRRSV-a u pokrajini Shandong, s lokalnih farmi prikupljeno je 1528 uzoraka svinja različitih kategorija za koje je postojala sumnja na zarazu PRRSVom. Kompletan genomski slijed soja SDLY-27 PRRSV-a određen je tehnologijom sekvenciranja sljedeće generacije (NGS). Slijed je imao dužinu od 15 363 nukleotida (nt), a komparativna analiza cijeloga genomskog slijeda uputila je na to da je homolognost između sojeva SDLY 27 i 81 PRRSV-a iz Kine i uzoraka u banci gena iz drugih zemalja 61,9~96,4%. Ovo je prvo istraživanje koje je otkrilo rekombinantne sojeve iz višestrukih rekombinacija među linijama 8 (sojevi nalik na JXA1), 5 (sojevi RespPRRSV-MLV i VR2332) i podlinije 1,5 (sojevi nalik na NADC34) u Shandongu, Kina.Kao takvo, istraživanje pruža nove podatke o epidemiologiji PRRSV-a u Kini, posebno u pokrajini Shandong, a ujedno predstavlja i važnu referenciju za razvoj novih cjepiva te prevenciju i kontrolu bolesti uzrokovane navedenim virusom

Potential molecular mechanisms of Erlongjiaonang action in idiopathic sudden hearing loss: A network pharmacology and molecular docking analyses

BackgroundIdiopathic sudden hearing loss (ISHL) is characterized by sudden unexplainable and unilateral hearing loss as a clinically emergent symptom. The use of the herb Erlongjiaonang (ELJN) in traditional Chinese medicine is known to effectively control and cure ISHL. This study explored the underlying molecular mechanisms using network pharmacology and molecular docking analyses.MethodThe Traditional Chinese Medicine System Pharmacological database and the Swiss Target Prediction database were searched for the identification of ELJN constituents and potential gene targets, respectively, while ISHL-related gene abnormality was assessed using the Online Mendelian Inheritance in Man and Gene Card databases. The interaction of ELJN gene targets with ISHL genes was obtained after these databases were cross-screened, and a drug component–intersecting target network was constructed, and the gene ontology (GO) terms, Kyoto Encyclopedia of Genes and Genomes, and protein–protein interaction networks were analyzed. Cytoscape software tools were used to map the active components–crossover target–signaling pathway network and screened targets were then validated by establishing molecular docking with the corresponding components.ResultErlongjiaonang contains 85 components and 250 corresponding gene targets, while ISHL has 714 disease-related targets, resulting in 66 cross-targets. The bioinformatical analyses revealed these 66 cross-targets, including isorhamnetin and formononetin on NOS3 expression, baicalein on AKT1 activity, and kaempferol and quercetin on NOS3 and AKT1 activity, as potential ELJN-induced anti-ISHL targets.ConclusionThis study uncovered potential ELJN gene targets and molecular signaling pathways in the control of ISHL, providing a molecular basis for further investigation of the anti-ISHL activity of ELJN

A universal optical modulator for synthetic topologically tuneable structured matter

Topologically structured matter, such as metasurfaces and metamaterials, have given rise to impressive photonic functionality, fuelling diverse applications from microscopy and holography to encryption and communication. Presently these solutions are limited by their largely static nature and preset functionality, hindering applications that demand dynamic photonic systems with reconfigurable topologies. Here we demonstrate a universal optical modulator that implements topologically tuneable structured matter as virtual pixels derived from cascading low functionality tuneable devices, altering the paradigm of phase and amplitude control to encompass arbitrary spatially varying retarders in a synthetic structured matter device. Our approach opens unprecedented functionality that is user-defined with high flexibility, allowing our synthetic structured matter to act as an information carrier, beam generator, analyser, and corrector, opening an exciting path to tuneable topologies of light and matter

Aberrant host immune response induced by highly virulent PRRSV identified by digital gene expression tag profiling

<p>Abstract</p> <p>Background</p> <p>There was a large scale outbreak of the highly pathogenic porcine reproductive and respiratory syndrome (PRRS) in China and Vietnam during 2006 and 2007 that resulted in unusually high morbidity and mortality among pigs of all ages. The mechanisms underlying the molecular pathogenesis of the highly virulent PRRS virus (H-PRRSV) remains unknown. Therefore, the relationship between pulmonary gene expression profiles after H-PRRSV infection and infection pathology were analyzed in this study using high-throughput deep sequencing and histopathology.</p> <p>Results</p> <p>H-PRRSV infection resulted in severe lung pathology. The results indicate that aberrant host innate immune responses to H-PRRSV and induction of an anti-apoptotic state could be responsible for the aggressive replication and dissemination of H-PRRSV. Prolific rapid replication of H-PRRSV could have triggered aberrant sustained expression of pro-inflammatory cytokines and chemokines leading to a markedly robust inflammatory response compounded by significant cell death and increased oxidative damage. The end result was severe tissue damage and high pathogenicity.</p> <p>Conclusions</p> <p>The systems analysis utilized in this study provides a comprehensive basis for better understanding the pathogenesis of H-PRRSV. Furthermore, it allows the genetic components involved in H-PRRSV resistance/susceptibility in swine populations to be identified.</p

C-Type Lectin in Chlamys farreri (CfLec-1) Mediating Immune Recognition and Opsonization

Background: C-type lectins are a superfamily of Ca 2+ dependent carbohydrate-recognition proteins that play significant diverse roles in nonself-recognition and clearance of invaders. Though they are well characterized in vertebrates, the study of the potential function and mechanism of C-type lectins in invertebrate immunity is still in its infancy. Methodology: A C-type lectin (CfLec-1) from scallop Chlamys farreri, a dominant cultured mollusk species in China, was selected to investigate its mRNA expression, localization and the possible functions in innate immunity in the present study. After scallop was stimulated by three typical PAMPs, the mRNA expression of CfLec-1 in hemocytes was poles apart. It was significantly up-regulated (p,0.01) after scallops were stimulated by LPS or b-glucan, but significantly down-regulated (p,0.01) after PGN stimulation. The binding ability of recombinant CfLec-1 (designated as rCfLec-1) towards eight PAMPs was investigated subsequently by PAMPs microarray, which revealed rCfLec-1 could bind LPS, PGN and mannan in vitro, indicating CfLec-1 served as a PRR involved in the pathogen recognition. Immunofluorescence assay with polyclonal antibody specific for CfLec-1 revealed that CfLec-1 was mainly located in the mantle and gill of the scallop. CfLec-1 could bind to the surface of scallop hemocytes and recruited hemocytes to enhance their encapsulation in vitro, and this process could be specifically blocked by anti-rCfLec-1 antibody. Meanwhile, rCfLec-1 could also enhance the phagocytic activity of scallop hemocytes against Escherichia coli

Understanding PRRSV Infection in Porcine Lung Based on Genome-Wide Transcriptome Response Identified by Deep Sequencing

Porcine reproductive and respiratory syndrome (PRRS) has been one of the most economically important diseases affecting swine industry worldwide and causes great economic losses each year. PRRS virus (PRRSV) replicates mainly in porcine alveolar macrophages (PAMs) and dendritic cells (DCs) and develops persistent infections, antibody-dependent enhancement (ADE), interstitial pneumonia and immunosuppression. But the molecular mechanisms of PRRSV infection still are poorly understood. Here we report on the first genome-wide host transcriptional responses to classical North American type PRRSV (N-PRRSV) strain CH 1a infection using Solexa/Illumina's digital gene expression (DGE) system, a tag-based high-throughput transcriptome sequencing method, and analyse systematically the relationship between pulmonary gene expression profiles after N-PRRSV infection and infection pathology. Our results suggest that N-PRRSV appeared to utilize multiple strategies for its replication and spread in infected pigs, including subverting host innate immune response, inducing an anti-apoptotic and anti-inflammatory state as well as developing ADE. Upregulation expression of virus-induced pro-inflammatory cytokines, chemokines, adhesion molecules and inflammatory enzymes and inflammatory cells, antibodies, complement activation were likely to result in the development of inflammatory responses during N-PRRSV infection processes. N-PRRSV-induced immunosuppression might be mediated by apoptosis of infected cells, which caused depletion of immune cells and induced an anti-inflammatory cytokine response in which they were unable to eradicate the primary infection. Our systems analysis will benefit for better understanding the molecular pathogenesis of N-PRRSV infection, developing novel antiviral therapies and identifying genetic components for swine resistance/susceptibility to PRRS

Evidence for cleavage of the metalloprotease Vsm from Vibrio splendidus strain JZ6 by an M20 peptidase (PepT-like protein) at low temperature

Metalloprotease Vsm is a major extracellular virulence factor of Vibrio splendidus. The toxicity of Vsm from V. splendidus strain JZ6 has been characterized, and production of this virulence factor proved to be temperature-regulated. The present study provides evidence that two forms (JZE1 and JZE2) of Vsm protein exist in extracellular products (ECPs) of strain JZ6, and a significant conversion of these two forms was detected by SDS-PAGE and immunoblotting analyses of samples obtained from cells grown at 4, 10, 16, 20, 24 and 28 ℃. Mass spectroscopy confirmed that JZE1 was composed only of the peptidase_M4 domain of Vsm, and JZE2 contained both the PepSY domain and the peptidase_M4 domain. An M20 peptidase T-like protein (PepTL) was screened from the transcriptome data of strain JZ6, which was considered as a crucial molecule to produce the active Vsm (JZE1) by cleavage of the propeptide. Similar to that of Vsm, PepTL mRNA accumulation was highest at 4 ℃ (836.82-fold of that at 28 ℃), decreased with increasing of temperature and reached its lowest level at 28 ℃. Deletion of the gene encoding the PepTL resulted in a mutant strain that did not produce the JZE1 cleavage product. The peptidase activity of PepTL recombinant protein (rPepTL) was confirmed by cleaving the Vsm in ECPs with an in vitro degradation reaction. These results demonstrate that PepTL participates in activating Vsm in strain JZ6 by proteolytic cleavage at low temperature