Molecular characterization and expression analysis of IL-22 and its two receptors genes in yellow catfish (Pelteobagrus filvidraco) in response to Edwardsiella ictaluri challenge.

Interleukin (IL)-22, as a member of the interleukin (IL)-10 family, is an important mediator between the immune cells and epithelial tissues during infection and inflammation. This study reported the characterization and mRNA expression patterns of Pf_IL-22 gene and its cell surface-associated receptors Pf_IL-22RA1 and soluble Pf_IL-22RA2 genes in yellow catfish (Pelteobagrus filvidraco). The open reading frames (ORFs) of the Pf_IL-22, Pf_IL-22RA1 and Pf_IL-22RA2 genes were 546 bp, 1740 bp and 690 bp in length, encoding 181, 579 and 229 amino acids, respectively. Alignments of the deduced amino acid sequences present that the Pf_IL-22 has a conserved IL-10 family signature motif, and the Pf_IL-22RA1 and Pf_IL-22RA2 have two conserved fibronectin type-III domains. Quantitative real-time PCR (qPCR) analyses showed that the Pf_IL-22 and Pf_IL-22RA1 mRNAs were highly expressed in mucosal tissues such as the fin, gill, intestine, skin mucus and stomach, and were weakly expressed in the kidney, liver and head kidney of adult yellow catfish, indicating that the Pf_IL-22 transcripts may be mainly produced by mucosal immune cells/tissues in healthy yellow catfish. The mRNA expression levels of the Pf_IL-22RA2 gene were high in the muscle and liver, and were relatively low in the spleen and kidney. The mRNA expression levels of the Pf_IL-22 and its two receptor genes were significantly up-regulated in both mucosal tissues (gill, hindgut, and skin mucus) and systemic immune tissues (spleen, head kidney and blood) after Edwardsiella ictaluri challenge. These results indicated that the Pf_IL-22 and its two receptors genes might play an important role in the innate immune defense against bacterial invasion.

The effect of magnetic coupling along the magnetic axis on slow magnetic relaxation in DyIII complexes with D5h configuration based on an aggregation-induced-emission-active ligand.

The adjustment of crystal symmetry and intramolecular magnetic coupling is of great importance for the construction of high-performance single-molecule magnets. By using an aggregation-induced-emission-active pyridine-carbohydrazone-based Schiff base ligand and phosphine oxides, four dinuclear and one one-dimensional DyIII-based complexes, [Dy2(TPE-pc)2(Bu3PO)2Cl2]·2CH3CN·2H2O (1), [Dy2(TPE-pc)2(Cy3PO)2Cl2] (2), [Dy2(TPE-pc)2(MePA)2Cl2]·2CH3OH (3), [Dy2(TPE-pc)2(Ph3PO)2Cl2]2 (4) and [Dy2(TPE-pc)2(DPPO)Cl2]n (5) (H2TPE-pc = (E)-N'-(2-hydroxy-5-(1,2,2-triphenylvinyl)benzylidene)picolinohydrazide, MePA = N-phenyl-N',N''-bis(morpholinyl) phosphoric triamide, DPPO = piperazine-1,4-diylbis(diphenyl phosphine oxide)), were isolated. All complexes are made up of an enol oxygen-bridged Dy2 unit, where DyIII ions possess a pentagonal bipyramidal geometry with pseudo D5h symmetry. Magnetic measurements reveal that intramolecular DyIII-DyIII couplings are ferromagnetic and all complexes display a significant slow magnetic relaxation phenomenon below 30 K under a zero dc field. Ab initio calculations indicate that the anisotropic magnetic axes of all DyIII ions are approximately perpendicular to the higher-order symmetric axes in all complexes, and that DyIII-DyIII magnetic couplings along the magnetic axes effectively suppress the ground state quantum tunneling effect of magnetization and promote the occurrence of slow magnetic relaxation. Raman relaxation prevails in all complexes. In addition, the H2TPE-pc ligand shows an aggregation-induced emission (AIE) effect; however, all complexes exhibit an aggregation-caused quenching (ACQ) phenomenon.

Effect of coordinated anions on ferromagnetically coupled Dy2 zero-field single-molecule magnets.

A new hydrazone Schiff base ligand was condensed from 2-hydroxy-3-methoxybenzaldehyde and pyrimidine-4-carbohydrazide {H2L = (E)-N'-(2-hydroxy-3-methoxybenzylidene)pyrimidine-4-carbohydrazide}, which was used to assemble two new Dy2 complexes Dy2L2(DMF)2(NO3)2 (1) and Dy2L2(DMF)2(AcO)2 (2). Notably, the coordinated anions have a subtle effect on the coordination configurations of the Dy3+ ions and the magnetic properties of the two Dy2 complexes. The Dy3+ ions in 1 and 2 have the same N2O5 coordination environment but show the triangular dodecahedron and the biaugmented trigonal prism coordination configurations, respectively. Magnetic measurements revealed that both 1 and 2 have intramolecular ferromagnetic interactions between the Dy3+ ions and show single-molecule magnet behaviors at 0 Oe, with Ueff/k values of 58.2 K for 1 and 59.9 K for 2. These magnetic properties may be explained by theoretical calculations.

Establishment and characterization of a fin cell line from blunt snout bream, Megalobrama amblycephala.

This study established and characterized a new cell line (MAF) from the fin of blunt snout bream (Megalobrama amblycephala), a freshwater fish cultivated in China. MAF cells proliferated well in medium 199 supplemented with 10 % fetal bovine serum at 28 °C and have been subcultured more than 95 times in almost a year. MAF cells were revived at 90-95 % viability after 3-6 months of storage in liquid nitrogen. Karyotyping indicated that the modal chromosome number of MAF cells was 48. The MAF cell line consisted predominantly of fibroblastic and epithelial-like cells from M. amblycephala, which was confirmed by immunofluorescence and mitochondrial 12s rRNA sequencing. Viral susceptibility tests showed that MAF cells were susceptible to infection by snakehead rhabdovirus, spring viremia carp virus, and channel catfish virus, which was demonstrated by the presence of cytopathic effect, high viral titers, and PCR products. Bacterial cytotoxicity studies showed that extracellular products from Aeromonas hydrophila were toxic to MAF cells. Cu²âº was also cytotoxic to MAF cells, and the 24-h IC50 value was 144.48 µmol/l. When MAF cells were transfected with pEGFP-N1 plasmid, bright fluorescent signals were observed, and the transfection efficiency reached up to 5 %. These results suggest that the MAF cell line may provide a valuable tool for studying virus pathogenesis, as well as cytotoxicity testing and genetic manipulation studies.

Value of UGT2B7-161 Single Nucleotide Polymorphism in Predicting the Risk of Cardiotoxicity in HER-2 Positive Breast Cancer Patients Who Underwent Pertuzumab Combined with Trastuzumab Therapy by PSL.

Objective: To explore the value of uridine diphosphate-glucuronosyltransferase 2B7 (UGT2B7)-161 single nucleotide polymorphism in predicting the occurrence of cardiotoxicity in Chinese human epidermal growth factor 2 (HER-2) positive breast cancer patients who underwent pertuzumab combined with trastuzumab Therapy. Methods: Fifty patients with HER-2 positive breast cancer who planned to receive trastuzumab and pertuzumab therapy for more than four cycles were enrolled in this study, and blood samples were collected. Thirty healthy volunteers of matching ages were selected as the control group. Myocardial parameters such as global work index, global effective work, and global wasted work were measured before treatment (M0) and at the end of four cycles of treatment month three (M3). Blood samples were collected from patients at the M0 stage, and polymorphisms of the UGT2B7-161 gene were detected to evaluate the predictive ability of different gene phenotypes on the myocardial drug toxicity injury. Results: There were 35 myocardial work decreased events among 50 patients. There were 24 (47.3%), 15 (40.8%), and 11 (11.8%) patients carrying UGT2B7-161 CC, CT, and TT genotypes, respectively. The occurrence of myocardial work decreased was decreased in UGT2B7-161 TT and CT genotypes (12.5%) compared with CC genotype (41.7%) with statistical significance (P < 0.001). Multivariate logistic regression model analysis exhibited that UGT2B7-161 genotypes, body mass index, and cardiac troponin I were independent factors influencing the risk of cardiotoxicity. Conclusion: UGT2B7-161 single nucleotide polymorphism is a potential predictive factor for cardiotoxicity in HER-2 positive breast cancer patients receiving trastuzumab and pertuzumab dual-targeted drug therapy.

Interleukin-9 promotes intestinal barrier injury of sepsis: a translational research.

BACKGROUND: Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. Intestinal mucosal barrier injury is one of the important manifestations of sepsis. Interleukin-9 (IL-9) and IL-9-producing CD4(+) T cells were emerging pro-inflammatory mediators with development of intestinal injury. However, it is unclear whether IL-9 is related to the intestinal barrier injury of sepsis. METHODS: To investigate the roles of IL-9-producing CD4(+) T cells and IL-9 in the process of barrier injury in sepsis, serum IL-9-producing CD4(+) T cell percentages, IL-9, and D-lactate levels were measured in septic patients and controls. The markers of barrier function in serum and intestinal tissue were also collected in septic rats. Moreover, the barrier injury degree and survival rate of septic rats were also investigated after increasing or interfering with IL-9 expression. RESULTS: The serum IL-9-producing CD4(+) T cell percentages, IL-9, and D-lactate levels were significantly higher in septic patients or rats than those in controls. IL-9-producing CD4(+) T cells and IL-9 levels were positively correlated with D-lactate levels and had a high predictive value of 28-day mortality in septic patients. The non-survivors had significantly higher serum T cell percentages, IL-9, and D-lactate levels compared with survivors. In septic rats, IL-9 increased the expression levels of D-lactate, whereas that decreased the expression levels of zonula occludens 1. Moreover, the barrier injury was aggravated or alleviated by increasing or interfering with IL-9 expression, respectively. Survival rate analysis also showed that IL-9 decreased the 14-day survival rate of septic rats. CONCLUSION: IL-9 is closely related to intestinal mucosal barrier injury and mortality in sepsis. IL-9 blockade has the potential to improve the barrier injury in sepsis. TRIAL REGISTRATION: The study was registered at ClinicalTrials.gov (ID: NCT03791866, Date: December 2018).

Risk factors for quinolone-resistant Escherichia coli infection: a systematic review and meta-analysis.

Background: Antimicrobial resistance to quinolone is rising worldwide, especially in Escherichia coli causing various infections. Although many studies have been conducted to identify the risk factors for quinolone-resistant Escherichia coli (QREC) infection, the results are inconsistent and have not been systematically reported. The aim of the present study is to conduct a systematic review and meta-analysis to evaluate the potential risk factors for QREC infection. Methods: A systematic search was performed to collect published data in the EMBASE, PubMed, and the Cochrane Library up to April 2019. Risk factors were analyzed using the pooled odds ratio (ORs) with 95% confidence interval (CIs). Results: Twenty-seven trials involving 67,019 participants were included in the present study. The following risk factors associated with QREC infection were identified: (1) male (OR = 1.41), (2) hepatic cirrhosis (OR = 2.05), (3) diabetes mellitus (OR = 1.62), (4) cardiovascular disease (OR = 1.76), (5) neurogenic bladder (OR = 8.66), (6) renal dysfunction (OR = 2.47), (7) transplantation (OR = 2.37), (8) urinary tract infection (OR = 2.79) and urinary tract abnormality (OR = 1.85), (9) dementia (OR = 5.83), (10) heart failure (OR = 5.63), (11) neurologic disease (OR = 2.80), (12) immunosuppressive drugs (OR = 2.02), (13) urinary catheter (OR = 4.39), (14) nursing home resident (OR = 4.63), (15) prior surgery (OR = 2.54), (16) quinolones (OR = 7.67), (17) other antibiotics (OR = 2.74), (18) hospitalization (OR = 2.06) and (19) nosocomial infection acquisition (OR = 2.35). Conclusions: QREC infection was associated with nineteen risk factors including prior quinolones use, hospitalization, and several comorbidities. Reducing exposure to these risk factors and modification of antibiotic use are important to prevent quinolone resistance.

Risk of Hepatocellular Carcinoma Remains High in Patients with HBV-Related Decompensated Cirrhosis and Long-Term Antiviral Therapy.

Background: This study aimed to evaluate the risk factors of HCC development in patients with hepatitis B virus (HBV)-related DC and who underwent long-term antiviral therapy. Methods: Data from 308 patients with HBV-related DC and long-term antiviral therapy were collected and retrospectively reviewed. Cox regression analysis was used to analyze independent risk factors of HCC development. Results: Data from 129 patients with definite records were analyzed. The median follow-up time was 5 years (range, 1 to 8 years). At the end of the follow-up, 41 (31.8%) patients developed HCC, and the time from DC diagnosis to HCC incidence who received antiviral therapy was 4.4 years (range, 1-7 years). The incidence of HCC was higher in males (30/78, 38.5%) than in females (11/51, 21.6%) (P = 0.04). Patients who developed HCC were significantly older than those who did not develop HCC (P < 0.01). The incidence of HCC in patients receiving nucleoside analogues, nucleotide analogues, and combination therapy was 34.7%, 38.1%, and 33.3%, respectively, and the difference showed no significant differences (P = 0.95). Multivariate Cox regression analysis demonstrated that male gender and age ≥50 years are independent risk factors of HCC development (OR = 2.987 and 2.408; 95% CI (1.301-6.858) and (1.126-5.149); P = 0.01 and 0.02, respectively). Conclusion: The risk of HCC remains to be high in patients with HBV-related DC, especially in males aged ≥50 years.

The molecular characterization, expression pattern and alternative initiation of Megalobrama amblycephala Hif prolyl hydroxylase Phd1.

HIF prolyl hydroxylase 1 (PHD1) functions in prolyl hydroxylation on mammal hypoxia-inducible factors (HIF), important transcription factors involved in hypoxia, however the roles of Phd1 in fish remain unclear. In this study, the full-length cDNA and promoter sequences of blunt snout bream (Megalobrama amblycephala) phd1 gene were isolated by a modified RACE strategy. The phd1 cDNA was 2672 bp for encoding 481 amino acid residues. In silico assays indicated that phd1 had 5 exons, and a 348 bp CpG island in the exon1, and several transcription factor binding sites (CAAT box, HRE, ARNT, FOX, etc) were also found on the promoter. The quantitative real-time PCR results suggested that phd1 mRNA was constitutively expressed in all detected tissues, and higher in the blood, brain and heart in normoxia, but significantly decreased after hypoxia in all detected tissues except for gill. Western blot assays indicated that two Phd1 isoforms were generated by alternative translation initiation. Moreover, these two isoforms were both localized in the nucleus, therein only the senior isoform promoted cell proliferation. Taken together, the present study firstly describes the functions of M. amblycephala two Phd1 isoforms in hypoxia and cell proliferation.

Molecular cloning and expression analysis of interleukin-1ß and interleukin-1 receptor type I genes in yellow catfish (Pelteobagrus fulvidraco): Responses to challenge of Edwardsiella ictaluri.

Interleukin-1ß (IL-1ß) is one of the pivotal early pro-inflammatory cytokines, which play important roles in regulating immune response and inducing a series of inflammatory reactions to infections. Interleukin-1 type I receptor (IL-1RI) is a receptor of the IL-1ß that can mediate IL-1-dependent activation. In this study, partial cDNA sequences of the Pf_IL-1ß and Pf_IL-1RI genes were cloned from yellow catfish (Pelteobagrus fulvidraco). The open reading frames (ORF) of Pf_IL-1ß and Pf_IL-1RI genes encode putative peptides of 280 and 543 amino acids, respectively. The deduced amino acid sequences of these two genes shared highly conserved structures with those from other teleosts. Quantitative real-time PCR results showed that the Pf_IL-1ß mRNA had relatively high expression levels in trunk kidney and blood, and the Pf_IL-1RI mRNA was highly expressed in blood and had relatively high expression level in liver. Ontogenetic expression analyses indicate that the Pf_IL-1ß and Pf_IL-1RI genes may play important roles during the embryonic developmental stages. The mRNA expression levels of Pf_IL-1ß and Pf_IL-1RI genes were up-regulated in the trunk kidney, head kidney, blood, spleen, heart and liver after Edwardsiella ictaluri challenge. Western blot analyses showed that Pf_IL-1ß protein was highly expressed in the spleen and head kidney, but not in the fin of adult individuals. These results suggest that the Pf_IL-1ß and Pf_IL-1RI genes may play significant roles in the immune regulation and defense against E. ictaluri in the yellow catfish.