Schizothorax prenanti (S. prenanti), an important species of economical fish in Southwest China, is susceptible to Aeromonas hydrophila (Ah). To understand the immune response to Ah, the ...transcriptome profiling of spleen of S. prenanti was analyzed after Ah infection. A total of 6, 213 different expression genes (DEGs) were obtained, including 3, 066 up-regulated DEGs and 3, 147 down-regulated DEGs. These DEGs were annotated by KEGG and GO databases, so that the immune-related DEGs (IRDs) can be identified and classified. Then, the interesting IRDs were screened to build heat map, and the reliability of the transcriptome data was validated by qPCR. In order to clarify the mechanism of signal transduction in the anti-bacterial immunity, the signaling pathway initiated by TLRs was predicted. In this pathway, TLR25 and TLR5 mediate the NF-κB and AP-1 signals via MyD88-dependent pathway. Meanwhile, the type I IFN (IFNα/β) induced by IRF1 and IRF3/7 may play an important role in the anti-bacterial immunity. In conclusion, this study preliminarily provides insights into the mechanism of signal transduction after Ah infection in S. prenanti, which contributes to exploring the complex anti-bacterial immunity.
•A series of immune-related DEGs were acquired by the transcriptome analysis.•TLR25 and TLR5 may mediate the NF-κB and AP-1 signals via MyD88-dependent pathway in the anti-bacterial immunity of fish.•The type I IFNs induced by IRFs play an important role in the anti-bacterial immunity of fish.•The transcriptome analysis reveals the functional difference of fish TLRs in anti-bacterial immunity.
The high performance and safety of lithium-sulfur (Li-S) batteries are the focus of research in recent years. The separator is an important part of the battery, whose performance stability directly ...affects the performance of the battery. What’s more, there are still many defects in the commercial polyolefin separator. In this work, a polyethylene/polyetherimide (PE/PEI) composite separator is designed by surface coating and non-solvent-induced phase separation (NIPS) method. The film breakage temperature and anti-puncture strength of the PE/PEI composite separator (181.96 °C, 7.90 N) are higher than those of the commercial PE separator (153.00 °C, 6.20 N), which will greatly ensure the safety problems caused by internal self-heating and external short circuit of Li-S batteries. The PE/PEI composite separator–based Li-S battery delivers a high initial specific capacity of 1265 mAh g
−1
at 0.2 C and exhibits excellent rate capability compared with the commercial PE separator. Therefore, the PE/PEI composite separator is a candidate separator for the high-performance and high-safety Li-S battery.
Follicle-stimulating hormone (FSH) has been newly demonstrated to play a great role in promoting fat accumulation, providing a potential to target FSH for controlling fat accumulation and treating ...obesity. A short, 13-amino acid of FSHβ (FSHβ13AA) was indentified to be the FSH receptor-binding epitope in both humans and mice. By conservation analysis, we found such FSHβ13AA is highly conserved across species. Accordingly, we designed a new FSH antigen by synthesizing a tandem of FSHβ13AA (LVYKDPARPNIQK) and then conjugating it to ovalbumin (FSHβ13AA-T-OVA). Then, we tested its efficacy in suppressing fat accumulation in both ovariectomized and intact mouse models. Vaccination with this novel antigen emulsified in mild adjuvant, Specol, was highly effective in preventing ovariectomy-induced body weight gain and fat accumulation in mice (P < 0.01). Mechanistically, FSH vaccination treatment inhibited lipid biosynthesis by inactivating PPARγ adipogenic signaling pathway and simultaneously enhanced adipocyte themogenesis via upregulating UCP1 expression in both visceral and subcutaneous adipose tissues. Moreover, injection of this novel FSH vaccine also substantially reduced (P < 0.05) fat accumulation in both intact male and female mice. These actions result from the specific binding of the generated antibody to the β-subunit to block its action, rather than lowering the circulating levels of FSH, as evidenced by nearly no alterations in serum FSH levels in mice following FSH vaccination. Overall, we developed a novel FSH antigen and vaccine, and demonstrated it is highly efficacious in suppressing fat accumulation.
•A novel FSH vaccine was developed based on the FSH receptor-binding epitope of FSH β-subunit.•Vaccination with this vaccine was highly effective in preventing ovariectomy-induced fat accumulation in mice.•Vaccination with this vaccine was also highly effective in preventing fat accumulation in intact mice fed on normal chow.•FSH vaccination suppresses fat accumulation through inactivating PPARγ and upregulating UCP1 signaling in adipose tissues.
Mammal Toll-like receptor 5 (TLR5) can directly recognize bacterial flagellin, initiate the inflammatory signaling cascades and trigger body immune system to clear the “non-self” substances. In ...teleosts, TLR5 has presented more complexes not only in increasing the molecular types, but also in elevating the functional diversity. In this study, we identified two TLR5 family members in Schizothorax prenanti, named as spTLR5-1 and spTLR5-2. The complete coding sequence (CDS) of spTLR5-1 is 2622 bp, encoding 873 amino acids, while the complete CDS of spTLR5-2 is 2640 bp, encoding 879 amino acids. Phylogenetic analysis showed that spTLR5-1 and spTLR5-2 were clustered to the TLR5 of schizothorax richardsonii and Cyprinus carpio respectively. The 3D structure analysis exhibited that the α-helix, β-sheet, and the ligand binding site of spTLR5-1, spTLR5-2 and human TLR5 have large differences. The spTLR5-1 and spTLR5-2 had extensively expressed in various tissues, including the higher expression in liver, spleen and head kidney. Both the expression levels of spTLR5-1 and spTLR5-2 were significantly up-regulated after Aeromonas hydrophila (A. hydrophila) challenge. And, the downstream genes, such as AP-1, IKK-α, NF-kB, IL-1β, IL-8 and TNF-α, were also significantly up-regulated after A. hydrophila challenge. Apart from that, the luciferase reporter assay demonstrated that the co-transfection of spTLR5-1 or spTLR5-2 into HEK293T cells showed the significantly increased NF-kB luciferase activity after flagellin stimulation. In conclusion, our results reveal that both two molecular types of fish TLR5 may commonly mediate the recognition of flagellin and the activation of the downstream inflammatory signaling molecules.
•spTLR5-1 and spTLR5-2 have similar structures and localize on the cell membrane.•Both spTLR5-1 and spTLR5-2 could respond to Aeromonas hydrophila challenge.•Both spTLR5-1 and spTLR5-2 could significantly activated NF-κB signaling pathway upon flagellin stimulation.
The molecular pathogenesis of myelodysplastic syndromes (MDS) has not been completely elucidated. Deregulation of expression of some microRNA has been implicated in hematological disorders including ...MDS. An oncogenic role for miR-22 was recently suggested in MDS in which the expression of miR-22 was increased. However, there were corroborations showing that it could be a tumor suppressor in acute myeloid leukemia. In this study, we examined the expression levels of circulating miR-22 in the plasma of patients with MDS and evaluated its significance in clinical context. The diagnosis of MDS was made according to the WHO classifications and subtypes included 8 cases of RA, 14 cases of RCMD and 6 cases of RARS/RCMD-RS, 10 cases RAEB1 and 7 cases RAEB2. Twenty cases of healthy donors were enrolled as normal control. Written informed consent for sample collection was obtained from all subjects enrolled. EDTA-anticoagulated peripheral blood samples were collected and centrifuged. Circulating microRNAs were purified from the plasma with miRNeasy serum kit (Qiagen). Expression of miR-22 was reverse- transcripted and measured with stem-loop real time quantitative polymerase chain reaction assay. First of all, we proved that the expression of miR-22 was detectable in the plasma of patients with MDS as well as of healthy donors. Our result showed that the expression levels of circulating miR-22 in MDS patients were higher than that in healthy donors (medians 1.360 vs 1.000, P<0.05), among which 13 patients (37.1%) showed marked increase of miR-22 expression (>2 times). Due to the high heterogeneity of MDS and relatively smaller sample size, the subtypes were grouped into two major categories based on IPSS: the low-risk group (including RA, RCMD and RARS/RCMD-RS, totally 18 cases) and the high-risk group (RAEB1 and RAEB2, 17 cases in total). Relative levels of circulating miR-22 were higher in the high-risk group than in the low-risk group (medians= 1.081 vs 2.317, P<0.05). A follow-up study revealed an association of the expression levels of miR-22 with prognosis. Patients with increased expression of miR-22 had poorer clinical response (Chi-square analysis, P<0.05) and lower overall survival rate (Kaplan-Meier analysis, P<0.05). We compared the levels of miR-22 in the plasma with that of mononuclear cells of bone marrow in some MDS patients and found that the relative levels of miR-22 in the plasma and bone marrow cells were not directly co-related (P>0.05), indicating it could be an independent prognostic factor. In summary, this study confirmed an oncogenic presence of miR-22 in MDS. Given the fact that there is a lack of blastic cells in the bone marrow of patients with low-risk MDS but their plasma or serum is easy to obtain, the detection of circulating miR-22 may be of value as a molecular marker in the prognosis of MDS.
No relevant conflicts of interest to declare.
The nucleotide-binding oligomerization domain 2 (NOD2) has been identified as an important sensor for microorganic invasion in both mammals and teleost fishes. In this study, two splicing variants of ...NOD2 (NOD2-v1 and NOD2-v2) were identified as truncating the functional domains of wild-type NOD2 in the teleost fish Schizothorax prenanti. NOD2-v1 included an intron sequence that terminated within the third leucine-rich repeat (LRR) domain, while NOD2-v2 incorporated an insertion of one and half intron sequences and truncated within the second caspase activation and recruitment domain (CARD). NOD2, NOD2-v1 and NOD2-v2 genes were ubiquitously expressed. All three genes positively responded to exposure of Aeromonas hydrophila and lipopolysaccharide stimulation in varying degrees. Using luciferase activity assays in HEK293T cells, our results revealed that NOD2 activated the NF-κB signal and recognized muramyl dipeptide (MDP). NOD2-v1 exhibited deficiency in the LRR domains and could not sense MDP, but maintained the ability to activate NF-κB and enhanced NOD2-mediated MDP recognition. Given the significant change to the functional structure, NOD2-v2 lost its capacity for NF-κB activation, but interestingly repressed NOD2-mediated MDP sensing and NF-κB activation, and even NOD2-v1-induced NF-κB activation. Altogether, our study reveals a novel pattern of signal regulation by splicing variants in teleost fishes.
•Human-like NOD2 splicing variants were firstly identified in S. prenanti, named NOD2, NOD2-v1 and NOD2-v2.•NOD2 and NOD2-v1 but NOD2-v2 can induce NF-κB activation.•NOD2-v1 and NOD2-v2 lose the function of sensing specific ligand MDP.•NOD2-v2 represses NOD2-mediated MDP sensing and NF-κB activation, and even NOD2-v1-induced NF-κB activation.
Quantification real-time PCR (qRT-PCR) is a common method in analysis of gene expression, but the stable reference genes for the normalization analysis have not been appreciated before identifying ...expression pattern of genes in teleost fishes. In this study, we selected eight candidate reference genes (18S, Actin, EF-1α, 40S, B2M, TUBA, UBCE and GAPDH) basing on transcriptome analysis and the traditional housekeeping genes, and analyzed the stability of the reference genes in spleen, head kidney and head kidney leukocytes (HKL) after pathogen challenge in Schizothorax prenanti (S. prenanti). Three common programs (geNorm, NormFinder and Bestkeeper) were used to evaluate the stability of the candidate reference genes. Two reference genes, Actin and EF-1α presented higher stability, while 18S and GAPDH were the lower stable genes, both in in vitro and in vivo. An important immune gene, toll-like receptor 22a (TLR22a), was selected to validate the stability of the proposed reference genes (Actin and EF-1α) across different experiment treatments. The results reveal that Actin and EF-1α are quite suitable reference genes for the normalization analysis. Otherwise, using the most stable gene Actin to validate the reliable of transcriptome data showed the high correlation between the fold change of transcriptome data and qRT-PCR data. In conclusion, our study not only acquired the suitable reference gene for the qRT-PCR assay under specific experiment condition, but also provided a comprehensive method to evaluate and validate the reference gene based on transcriptome analysis in teleost fishes.
•The stable reference genes were firstly analyzed in Schizothorax prenanti after pathogen challenge, based on RNA-seq.•Actin and EF-1α were considered as the suitable reference genes, stability: Actin > EF-1α.•Our study provided a comprehensive method to evaluate and validate the reference gene based on transcriptome analysis in teleost fishes.
Enterovirus 71 (EV-A71) and Coxsackievirus A16 (CV-A16) are the two most common pathogens causing hand, foot, and mouth disease (HFMD). Previously, we obtained one candidate live attenuated strain ...each for EV-A71 and CV-A16; here, we evaluated the safety and immunogenicity of a combinedlive vaccine against EV-A71 and CV-A16 generated from these two candidate strains. Rhesus monkeys were intramuscularly treated with a live combinationvaccine against both EV-A71 and CV-A16 or with either vaccine alone. No fever or atypical clinical signs were observed in any animals. Monkeys vaccinated with the combinationlive vaccine presented no notable pathological changes in the brain, spinal cord, lung, and liver; in contrast, these regions showed inflammatory cell infiltration in monkeys treated with EV-A71 alone or CV-A16 alone. Weak viremia was detected in plasma after inoculation with the combinationvaccine; however, the duration of viral shedding in feces was increased. Biochemical studies revealed a slight increase in aspartate aminotransferase levels in monkeys inoculated with the live combination vaccine; however, histopathological findings did not attribute this change to liver damage. We also found that the live combinationvaccine induced a dual humoral immune response. Cytokine analysis indicated that the combined EV-A71/CV-A16 vaccine significantly down-regulated interleukin-8 production. Here, we have demonstrated that the live attenuated EV-A71/CV-A16 vaccine was safe and could trigger a dual specific immune response. However, its immune protection efficacy requires further investigation.
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