Plant microRNAs (miRNAs), a class of about 21-nucleotide-long small noncoding RNAs (ncRNAs), generally act as key regulators of their target genes by guiding mRNA cleavage or translational ...repression. Recent researches have reported that miRNAs can interact with pathway-related structural genes, transcription factors (TFs) and noncoding RNAs (ncRNAs) to form regulatory network in secondary metabolite (SM) biosynthetic pathways. The validated interactions can better reflect the real regulatory mechanism of miRNA and convert miRNAs into more efficient tools to control the production of precious SMs. However, there is no systematic review available on this topic especially in plants, particularly model plants and crops. Here, we firstly overviewed the critical secondary metabolic pathways in plants, especially those biosynthetic pathways related structural genes which were well-studied and representative SMs, including phenylpropanoids, terpenoids, alkaloids. Principally, we summarized miRNAs involved in the biosynthesis of SMs, miRNA-target modules and their regulation on the mentioned pathways in the last decade. Significantly, the modules included miRNA-structural gene, miRNA-TF, miRNA-ncRNA interaction pairs carried out by target validation or functional confirmation. This knowledge will promote understanding the sophisticated miRNA-mediated gene regulatory network of SM biosynthesis, and drive the development of synthetic biology.
•TLR13a and TLR13b genes were identified from tilapia (Oreochromis niloticus).•Expressions of TLR13a and TLR13b mRNAs were altered after Streptococcus agalactiae challenge.•The TLR13a and TLR13b ...mRNAs were detected in various embryonic developmental stages.•Both OnTLR13a and OnTLR13b could interact with OnMyd88.•Both OnTLR13a and TLR13b can combine with S. agalactiae 23S rRNA.
Toll-like receptors (TLRs) play a crucial role in the innate immune system, which is the first line of defence against pathogens and pathogenic products in fish. In the present study, we cloned the full-length cDNA and genome sequences of two TLR13 s (OnTLR13a, OnTLR13b) from Nile tilapia (Oreochromis niloticus). TLR family motifs, i.e., the leucine-rich repeat (LRR) domains and Toll/interleukin (IL)-1 receptor (TIR) domains, were conserved in the putative proteins OnTLR13a and OnTLR13b, with fifteen LRR domains and one TIR domain. Four exons and three introns were identified in the OnTLR13a genome sequence, and three exons and two introns were identified in the OnTLR13b genome sequence. In healthy Nile tilapia tissues, OnTLR13a and OnTLR13b were ubiquitously expressed in all 11 tested tissues/organs. The highest expression levels were observed in the spleen (OnTLR13a) and blood (OnTLR13b), and the lowest expression levels were observed in the liver (OnTLR13a) and stomach (OnTLR13b). The expression level of OnTLR13b at 5.5 days postfertilization (dpf) was significantly higher than that at the other 8 time points (2.5, 3.5, 4.5, 5, 6, 6.5, 7.5 and 8.5 dpf). Upon stimulation with an intraperitoneal injection of 200 μL (107 CFU/mL) Streptococcus agalactiae, the expression levels of OnTLR13a and OnTLR13b were significantly upregulated in the intestine and gill. After cotransfection with MyD88, OnTLR13a significantly increased MyD88-dependent NF-κB activation in 293 T cells. However, OnTLR13b significantly impaired MyD88-dependent NF-κB activation. In addition, TLR13a slightly increased MyD88-dependent AP-1 activation, and TLR13b significantly increased MyD88-dependent AP-1 activation. TLR13a significantly increased MyD88-dependent interferon-β (IFN-β) activation, and TLR13b had no effect on MyD88-dependent IFN-β activation. These findings suggest that although the deduced protein structure of OnTLR13 is evolutionarily conserved between OnTLR13 and other TLR members, its signal transduction function is markedly different. Co-immunoprecipitation (Co-IP) assays showed that both OnTLR13a and OnTLR13b could interact with OnMyD88. RNA pulldown assays showed that TLR13a and TLR13b could combine with the 23S rRNA of S. agalactiae. These results indicate that TLR13a and TLR13b play important roles in the innate immune response against bacterial infection in Nile tilapia.
Tripartite motif (TRIM) proteins play a regulatory function in cancer, cell apoptosis and innate immunity. To understand the role of TRIM39 in Nile tilapia (Oreochromis niloticus), TRIM39 cDNA was ...isolated. The total length of TRIM39 cDNA was 5025 bp. The deduced OnTRIM39 protein contains 549 amino acids and has conserved domains of the TRIM family, which are the RING, B‐box, coiled‐coil and PRY‐SPRY domains. OnTRIM39 mRNA was widely expressed in various tissues. After challenge with Streptococcus agalactiae and stimulation with polyinosinic polycytidylic acid poly (I:C) and lipopolysaccharides (LPS), the amount of OnTRIM39 transcript was changed in various tested tissues. OnTRIM39 overexpression increased NF‐κB activity. OnTRIM39 was present in the cytoplasm. Mass spectrometry of proteins pulled down with recombinant OnTRIM39 showed that 250 proteins potentially interact with OnTRIM39. The authors selected I3K4I3 from the 250 candidate proteins to verify its interaction with TRIM39. They also selected I3KL45, a member of the same 14‐3‐3 protein family, to verify its interaction with TRIM39. The results of pull‐down assays showed that OnTRIM39 interacted with both I3K413 and I3KL45. These results contribute to further study of the innate immune mechanism of tilapia.
The nucleotide-binding oligomerization domain proteins NOD1, NOD2 and NLRC3 are cytoplasmic pattern recognition receptors (PRRs) of the Nod-like receptor (NLR) family. In the present study, the Nile ...tilapia (Oreochromis niloticus) NOD1 (ntNOD1), NOD2 (ntNOD2) and NLRC3 (ntNLRC3) genes were cloned and characterized. The full-length ntNOD1, ntNOD2 and ntNLRC3 genes were 3924, 3886 and 4574 bp, encoding 941, 986 and 1130 amino acids, respectively. The three Nod-like receptors have a NACHT domain and a C-terminal leucine-rich repeat (LRR) domain. In addition, ntNOD1 and ntNOD2 have a N-terminal CARD domain (ntNOD2 has two). Phylogenetic analysis showed that the three NLRs are highly conserved. Tissue expression analysis of the three receptors revealed that the highest mRNA and protein levels of ntNOD1, ntNOD2 and ntNLRC3 were in the spleen. The expression patterns of NLRs during embryonic development showed that the expression levels of ntNOD2 and ntNLRC3 significantly increased from 2 to 8 days post-fertilization (dpf). The expression levels of ntNOD1 significantly increased from 2 to 6 dpf, decreased at 7 dpf and then increased at 8 dpf. Upon stimulation with an intraperitoneal injection of Streptococcus agalactiae, expression levels of the ntNOD1, ntNOD2 and ntNLRC3 mRNA and protein were clearly altered in the blood, spleen, kidney, intestine and gill. Furthermore, after cotransfection with an NF-κB reporter plasmid, NF-κB activation in ntNOD1-overexpressing 293T cells significantly increased compared with that in control cells, before or after i-EDPA-stimulation. By contrast, compared with control, ntNOD2 and ntNLRC3 had no effect on NF-κB activation in 293T cells, when their potential ligands were not stimulated. However, after MDP-stimulation, ntNOD2 and ntNLRC3 overexpression increased NF-κB activation in 293T cells. NOD1 and NLRC3 were uniformly distributed throughout the cytoplasm in 293T cells, whereas NOD2 was distributed throughout the cytoplasm and nucleus. Our results indicate that the three Nod-like receptors are functionally conserved and may play pivotal roles in defense against pathogens such as Streptococcus agalactiae.
•NOD1, NOD2 and NLRC3 cDNAs were isolated from tilapia Oreochromis niloticus.•NOD1, NOD2 and NLRC3 mRNAs and proteins expresses ubiquitously with predominant expression in spleen.•Expressions of NOD1, NOD2 and NLRC3 mRNAs and proteins were up-regulated after Streptococcus agalactiae challenge.•The ntNOD1, ntNOD2, ntNLRC3 mRNAs were detected in various embryonic developmental stages.•The ntNOD1, ntNOD2 and ntNLRC3 genes were positively involved in the NF-κB-mediated signaling pathway.
In this study, we cloned the complementary (c)DNA sequences of tumour necrosis factor receptor (TNFR)‐associated factor 3 (traf3) in Nile tilapia, Oreochromis niloticus. The expression patterns of ...the traf3 gene were investigated and preliminary functional analyses were performed. In healthy fish, traf3 transcript was broadly expressed in all examined tissues, with the highest expression level in the blood and the lowest in the liver. The traf3 gene reached its highest expression at 8 days post‐fertilisation (dpf) during embryonic development. Moreover, we found that expression of traf3 was clearly altered following stimulation with Streptococcus agalactiae in vivo and that traf3 could be induced by lipopolysaccharides (LPS), Poly I: C and S. agalactiae WC1535 in Nile tilapia macrophages. Overexpression in 293T cells showed that Traf3 protein was mainly distributed in the cytoplasm and could significantly increase nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) activation. Taken together, these results implied that traf3 could play important roles in the immune response to pathogen invasion.
The recognition of microbial pathogens, which is mediated by pattern recognition receptors (PRRs), is critical to the initiation of innate immune responses. In the present study, we isolated the ...full-length cDNA and genomic DNA sequences of the MDA5, LGP2 and MAVS genes in Nile tilapia, termed OnMDA5, OnLGP2 and OnMAVS. The OnMDA5 gene encodes 974 amino acids and contains two caspase-associated recruitment domains (CARDs), a DExDc domain (DExD/H box-containing domain), a HELICc (helicase superfamily C-terminal) domain and a C-terminal regulatory domain (RD). The OnLGP2 gene encodes 679 amino acids and contains a DExDc, a HELICc and an RD. The OnMAVS gene encodes 556 amino acids and contains a CARD, a proline-rich domain, a transmembrane helix domain and a putative TRAF2-binding motif (269PVQDT273). Phylogenetic analyses showed that all three genes from Nile tilapia were clustered together with their counterparts from other teleost fishes. Real-time PCR analyses showed that all three genes were constitutively expressed in all examined tissues in Nile tilapia. OnMDA5 presented the highest expression level in the blood and the lowest expression level in the liver, while OnMAVS presented the highest expression level in the kidney. The highest expression level of OnLGP2 was detected in the liver. An examination of the expression patterns of these RIG-I-like receptors (RLRs) during embryonic development showed that the highest expression levels of OnMDA5 occurred at 2 days postfertilization (dpf), and the expression significantly decreased from 3 to 8 dpf. The expression levels of OnLGP2 significantly increased from 4 to 8 dpf. The expression levels of OnMAVS mRNA were stable from 2 to 8 dpf. Upon stimulation by intraperitoneal injection of Streptococcus agalactiae, the expression levels of OnMDA5 were first downregulated and then upregulated in the blood, gill and spleen. In the intestine and kidney, the expression of OnMDA5 was first upregulated, then downregulated, and then upregulated again. The expression of OnLGP2 was upregulated in the kidney and intestine, and the expression of OnMAVS was upregulated in the spleen. Overexpression of OnMAVS increased NF-κB activation in 293 T cells (p < 0.05), and after cotransfection with OnMDA5, the OnMAVS-dependent NF-κB activation was slightly increased (p > 0.05), after cotransfection with OnLGP2, the OnMAVS-dependent NF-κB activation was significantly decreased (p < 0.05). These findings suggest that, although the deduced protein structure of OnMDA5 is evolutionarily conserved with the structures of other RLR members, its signal transduction function is markedly different. The results also suggest that OnLGP2 has a negative regulatory effect on the OnMAVS gene. OnMDA5 and OnMAVS were uniformly distributed throughout the cytoplasm in 293 T cells, whereas OnLGP2 was distributed throughout the cytoplasm and nucleus. These results are helpful for clarifying the innate immune response against bacterial infection in Nile tilapia.
•MDA5, LGP2 and MAVS genes were isolated from tilapia Oreochromis niloticus.•MDA5, LGP2 and MAVS mRNAs expressed ubiquitously with predominant expression in spleen.•Expressions of MDA5, LGP2 and MAVS mRNAs were altered after Streptococcus agalactiae challenge.•The MDA5, LGP2 and MAVS mRNAs were detected in various embryonic developmental stages.•OnLGP2 could significantly decrease the OnMAVS-dependent NF-κB activation.
The association between major histocompatibility complex (MHC) class IIA polymorphisms and the severity of infection by Streptococcus agalactiae was investigated using 40 susceptible and 40 resistant ...individuals of Nile tilapia Oreochromis niloticus. Twenty‐five alleles were identified from 80 individuals, which belong to 22 major allele types. High polymorphism of mhcIIa gene and at least two loci were discovered in O. niloticus. In peptide‐binding region (PBR) and non‐PBR, the ratio of nonsynonymous substitution (dN) to synonymous substitution (dS) was 1.294 (>1) and 1.240 (>1), suggesting that the loci are evolving under positive balancing selection. Association analysis showed that the allele, orni‐daa*0501, was significantly associated with resistance to S. agalactiae, while the alleles, orni‐daa*1101, orni‐daa*1301, orni‐daa*1401 and orni‐daa*1201, were associated with susceptibility to S. agalactiae. To confirm these correlations, another independent challenge experiment was performed in the Huizhou population of the O. niloticus. The frequency distribution showed that the orni‐daa*1101 allele was significantly more frequent in the Huizhou‐Susceptible group (HZ‐SG) than in the Huizhou‐Resistant group (HZ‐RG) (P < 0.05), which was consistent with the first challenge. However, orni‐daa*0501 did not present in HZ‐SG and HZ‐RG and the distribution frequencies of the orni‐daa*1201, orni‐daa*1301 and orni‐daa*1401 alleles were not significantly more frequent in HZ‐SG than in HZ‐RG. These results indicate that the orni‐daa*1101 allele confers susceptibility to S. agalactia infection. These results suggest that the diversity of exon 2 of mcaIIa alleles could be used to explore the association between disease susceptibility or resistance and the multiformity of mcaIIa and to achieve the molecular‐assisted selection of O. niloticus with enhanced disease resistance.
The traditional Chinese medicine classic herbal formula
decoction (LDXGD) is widely used for hypertensive patients in China.
This systematic review and meta-analysis aimed to evaluate the efficacy ...and safety of LDXGD for hypertension.
PubMed, EMBASE, Cochrane Central Register of Controlled Trials, Chinese Biomedical Literature Database, Chinese Scientific Journal Database, Chinese National Knowledge Infrastructure, and Wanfang Database were searched up to February 7, 2017 for randomized control trials in treating hypertension.
Nine trials were identified. Compared with antihypertensive drugs, Longdanxiegan decoction plus antihypertensive drugs (LPAD) significantly improved systolic blood pressure (BP) (
= 138; MD = -4.82 mmHg; 95% CI: -7.89 to -1.76;
= 0.002), diastolic BP (
= 138; MD = -2.42 mmHg; 95% CI: -3.22 to -1.62;
< 0.00001), categorical BP (
= 509; RR: 1.26; 95% CI: 1.17 to 1.36;
< 0.00001), hypertension related symptoms (
= 509; RR: 1.31; 95% CI: 1.15 to 1.49;
< 0.0001), and heart rate (
= 138; MD = -2.40 bpm; 95% CI: -4.23 to -0.56;
= 0.01). Beneficial effects but no statistically significant reduction in total cholesterol (
= 138; MD = -0.11 mmol/l; 95% CI: -0.65 to 0.44;
= 0.71), or triglyceride (
= 138; MD = -0.20 mmol/l; 95% CI: -0.46 to 0.07;
= 0.14) was observed in LPAD. Compared with antihypertensive drugs, LDXGD used alone significantly improved systolic BP, diastolic BP, and hypertension related symptoms. But there was no difference between LDXGD and antihypertensive drugs on categorical BP (
= 120; RR: 1.09; 95% CI: 0.96 to 1.23;
= 0.18). The safety of LDXGD were still unclear.
Due to poor methodological quality of the included trials, as well as potential reporting bias, our review found no conclusive evidence for the effectiveness of LDXGD in treating hypertension. The potential beneficial effects and safety of LDXGD should be assessed in future properly designed trials.
Background
The impact of hepatic venous anatomic variations on hepatic resection and transplantation is the least understood aspect of liver surgery.
Methods
A prospective three-dimensional computed ...tomography study was undertaken on 200 consecutive subjects with normal livers to determine the prevalence of surgically significant hepatic venous anatomic variations.
Results
The prevailing pattern of the three hepatic veins in these subjects was a right hepatic vein (RHV) and a common trunk for the middle (MHV) and left (LHV) hepatic veins (122/200, 61%). The remaining patients had the RHV, MHV, and LHV draining independently into the inferior vena cava (IVC). In 39% of patients, the RHV was small and was compensated by a large right inferior hepatic vein (21.0%), an accessory RHV (8.5%) or a well-developed MHV (6.5%). A segment 4 vein was seen in 51.5% of patients. This segment 4 vein joined the LHV (26%), the MHV (17.5%), or the IVC (8%). An umbilical vein and a segment 4 vein were seen in 3.5% of patients. These two veins joined either the LHV (2.0%) or the MHV (1.5%).
Conclusions
Knowing the variations of hepatic veins before surgery is useful during both partial hepatectomy and donor operations for living related liver transplantation.
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