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1.
Mol Immunol ; 133: 86-94, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33636433

RESUMO

Regulatory B cells (Bregs), a subset of B lymphocytes discovered in the past few decades, have the capacity to suppress the immune response and dampen inflammation by secreting cytokines (IL-10 and TGF-ß). Whether Bregs are involved in Trichinella spiralis infection and the phenotypic characteristics of these cells after infection are still unknown. We investigated the phenotype of and dynamic changes in IL-10-producing Bregs in Trichinella spiralis infection in BALB/c mice. We used multicolour fluorescence immunostaining of microwave-treated paraffin sections to investigate the number of Bregs in T. spiralis infection. Flow cytometry (FCM) was used to determine the frequency of Bregs and related subgroups and cytokines in the spleen and mesenteric lymph nodes (MLNs). High levels of IL-10 were detected in the spleen and MLNs of mice after infection with T. spiralis. Furthermore, the frequencies of IL-10-producing CD19+CD1dhighCD5+ regulatory B cells and CD19+ cells were increased during T. spiralis infection. We also showed that the induced phenotype was similar to that of transitional type 2 marginal zone precursor B cells (T-MZP) cells after T. spiralis infection in mice. This study is the first demonstration of the expansion of Bregs following T. spiralis infection.

2.
J Clin Lab Anal ; : e23722, 2021 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-33543801

RESUMO

BACKGROUND: Myasthenia gravis (MG) is an autoimmune disease mediated by acetylcholine receptor antibodies. Exosomes were shown to be involved in the immune modulation and autoimmune diseases. However, the expression and function of exosomal long noncoding RNAs (lncRNAs) in MG are still unclear. METHODS: We conducted high-throughput sequencing to detect the lncRNA profiles of serum exosomes in 6 MG patients (2 grade I, 2 grade IIa, and 2 grade IIb) and 6 healthy controls (HC). Then, differentially expressed (DE) lncRNAs with the greatest difference between the MG and HC groups were selected for further quantitative real-time polymerase chain reaction (qRT-PCR) validation in additional 30 MG patients and 10 HC. The DE lncRNAs were used to construct the coding/noncoding network and perform enrichment analysis. RESULTS: We identified 378 significantly upregulated and 348 significantly downregulated lncRNAs in MG patients compared with HC. The top 5 lncRNAs (NR_104677.1, ENST00000583253.1, NR_046098.1, NR_022008.1, and ENST00000581362.1) were validated and shown to be significantly increased in the serum exosome of MG, and the expression level of NR_046098.1 significantly increased with the MG grading. Enrichment analysis showed that DE genes mainly participated in the basic biological regulation of MG and immune-related pathways, such as autoimmune thyroid disease pathway and T-cell receptor signaling pathway. A specific lncRNA-miRNA-mRNA regulatory network associated with the 5 lncRNAs, 14 MG-related miRNAs and 30 mRNAs was constructed. CONCLUSIONS: We conducted a comprehensive analysis of exosomal lncRNAs to reveal potential biomarkers for the MG diagnosis and severity assessment.

3.
Cancer Manag Res ; 12: 11005-11014, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33173334

RESUMO

Aim: This study aimed to investigate the effect of long-chain non-coding SOX21-AS1 on the proliferation and migration of breast cancer (BC) cells through the PI3K/AKT signaling pathway. Methods: Eighty-eight BC and adjacent tissues were collected, and BC cells and normal breast epithelial cells were purchased. LncRNA SOX21-AS1 expression in tissues and cells was detected by RT-PCR. miR-NC, si-SOX21-AS1, and Sh-SOX21-AS1 were transfected into BC cells. The PI3K/AKT signaling pathway was interfered with L740Y-P (activator of the PI3K/AKT pathway) and LY294002 (inhibitor of the PI3K/AKT pathway) in BC cells. The SOX21-AS1 expression in BC tissues and cells was tested by qRT-PCR, and the expression levels of p-PI3K, p-AKT, N-cadherin, E-cadherin, and vimentin were detected by WB. Results: SOX21-AS1 was highly expressed in BC, and the p-PI3K and p-AKT levels were also high. Cell experiments showed that inhibiting SOX21-AS1 expression could inhibit the proliferation, invasion, migration, and EMT of BC cells, and up-regulating its expression could promote the proliferation, invasion, migration, and EMT of ovarian cancer cells. The tumor-forming experiment in nude mice was consistent with the results in vitro. 740Y-P intervention could reverse the inhibition effect of Si-SOX21-AS1 on BC cell proliferation, invasion, migration, and EMT, while LY294002 intervention could reverse the promotion effect of Sh-SOX21-AS1 on BC cell proliferation, invasion, migration, and EMT. Conclusion: SOX21-AS1 is highly expressed in BC tissues. Silencing BC expression can inhibit the proliferation, invasion, migration, and EMT of cells by inhibiting the PI3K/AKT signaling pathway, which may be a new target for diagnosis and treatment.

4.
Vet Microbiol ; 249: 108834, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32919197

RESUMO

The H9N2 avian influenza virus (AIV) causes serious economic losses to the poultry industry every year. Vaccines that induce a mucosal immune response may be successful against influenza virus infection because its transmission occurs primarily in the mucosa. To develop novel and potent oral vaccines based on Lactobacillus plantarum (L. plantarum) to control the spread of AIV in poultry industry, in the present study, we constructed and expressed fusions of the influenza antigens NP and M2 with the Salmonella Typhimurium flagellinprotein FliC on the surface of L. plantarum. Oral immunization of chicks was performed, and serum antibodies, mucosal antibodies, and specific cellular immunity were detected. Immunizing chicks with avian influenza virus was evaluated. The results showed high levels of IgG in addition to high levels of secretory immunoglobulin A (sIgA) in chickens orally administered recombinant L. plantarum. In addition, the fusion may significantly increase the levels of NP- and M2-specific T cell-mediated immunity in the case of mucosal administration of NC8-pSIP409-pgsA'-NP-M2-FliC. Recombinant NC8-pSIP409-pgsA'-NP-M2-FliC mediated effectively protected chickens against influenza virus and reduced virus titers in the lung. Our study outcomes indicate that the expression of influenza NP-M2 and a mucosal adjuvant (FliC), by L. plantarum could generate a mucosal vaccine candidate for animals in the future to defend against AIVs.

5.
Parasitol Res ; 119(9): 2885-2895, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32715344

RESUMO

Chicken coccidiosis is a protozoan parasitic disease that leads to considerable economic losses in the poultry industry. In this study, we used invasive Lactobacillus plantarum (L.P) expressing the FnBPA protein as a novel bacterial carrier for DNA delivery into epithelial cells to develop a live oral DNA vaccine. A fusion DNA vaccine co-expressing EtMIC2 and chicken IL-18 (chIL-18) was constructed and then delivered to the host by invasive L.P. Its efficacy against Eimeria tenella challenge was evaluated in chickens by examining the relative weight gain rate; caecal lesion score; OPG; anti-coccidial index (ACI); levels of EtMIC2 antibody, FnBPA, IL-4, IL-18, IFN-γ and SIgA; and proliferation ability and percentages of CD4+ and CD8+ splenocytes. The experimental results showed that chickens immunized with invasive L.P carrying the eukaryotic expression vector pValac-EtMIC2 (pValac-EtMIC2/pSIP409-FnBPA) had markedly improved immune protection against challenge compared with that of chickens immunized with non-invasive L.P (pValac-EtMIC2/pSIP409). However, invasive L.P co-expressing EtMIC2 with the chIL-18 vector exhibited the highest protection efficiency against E. tenella. These results indicate that invasive Lactobacillus-expressing FnBPA improved humoural and cellular immunity and enhanced resistance to E. tenella. The DNA vaccine delivered by invasive Lactobacillus provides a new concept and method for the prevention of E. tenella.


Assuntos
Antígeno 12E7/metabolismo , Coccidiose/veterinária , Eimeria tenella/imunologia , Interleucina-18/metabolismo , Lactobacillus plantarum/metabolismo , Vacinas Protozoárias/imunologia , Vacinas de DNA/imunologia , Animais , Ceco/parasitologia , Galinhas/parasitologia , Coccidiose/parasitologia , Eimeria tenella/genética , Imunidade Celular/imunologia , Imunoglobulina A Secretora/genética , Lactobacillus plantarum/genética , Doenças das Aves Domésticas/parasitologia , Doenças das Aves Domésticas/prevenção & controle , Vacinação/veterinária , Ganho de Peso
7.
Vet Parasitol ; 283: 109161, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32526607

RESUMO

Coccidiosis is an intestinal parasitic disease that is caused by Eimeria tenella and other species, and it seriously restricts the economic development of the broiler breeding industry. In this study, a recombinant Lactobacillus plantarum with an invasive effect was constructed, and it expressed the TA4-AMA1 protein of E. tenella. After oral immunization with recombinant L. plantarum, specific humoral and mucosal immune levels were measured by indirect ELISA, and the differentiation of T cells was analysed by flow cytometry. After challenge with sporulated oocysts, the body weight, oocyst shedding and cecum lesions of the chicken were evaluated. The results indicated that chickens immunized with recombinant invasive L. plantarum produced higher levels of specific antibodies in the serum than did the non-immunized controls, and the secretory IgA (sIgA) levels were increased in the intestinal washes compared to those of the controls (P < 0.05). Flow cytometry showed that recombinant invasive L. plantarum significantly stimulated T cell differentiation compared to the PBS group (P < 0.01, P < 0.001), and a higher proportion of CD4+ and CD8+ T cells were detected in peripheral blood. Moreover, the lesion scores and histopathological caecum sections showed that immunizing chickens with recombinant invasive L. plantarum can significantly relieve pathological damage in the cecum (P < 0.01), and the relative body weight gain was 89.64 %, which was higher than the 79.83 % gain in the chickens immunized with non-invasive L. plantarum. After the challenge, faeces from ten chickens in each group were collected between 4 and 7 days, and the oocysts per gram (OPG) was determined by the McMaster technique. The data indicated that oocysts in the faeces of chickens immunized with the recombinant invasive L. plantarum were significantly lower than those of the controls (P < 0.01). The results suggest that recombinant invasive L. plantarum effectively activated immune responses against E. tenella infection and can be used as a candidate vaccine against E. tenella infection.

8.
J Air Waste Manag Assoc ; 70(9): 904-914, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32412866

RESUMO

In this study, an effective circulating system was developed to remove heavy metals in medical waste incineration (MWI) fly ash. MWI fly ash (MWIFA)-column experiments were performed to remove Cu, Pb, Zn, Cd, and Ni from MWIFA using EDTA disodium (Na2EDTA). Iron-column experiments were conducted to study the removal effect of zero-valent iron on the five heavy metals from washing wastewater. Toxicity Characteristic Leaching Procedure (TCLP) test method was employed to evaluate heavy metals toxicity of MWIFA residues generated after 0-0.2 mol/L Na2EDTA solution treated. After being washed by 0.2 mol/L Na2EDTA solution, TCLP leaching values of Cu, Pb, Zn, Cd, and Ni were the lowest and satisfied the standard (GB 5085.3-2007), and the leaching values were 58.4 ± 2.0 mg/L, 2.81 ± 0.14 mg/L, 64.3 ± 4.0 mg/L, 0.156 ± 0.005 mg/L, 0.381 ± 0.006 mg/L. Concentrations of Cu, Pb, Zn, Cd, and Ni in iron-column effluent were reduced by 99.7%, 91.6%, 91.6%, 75.4%, and 75.7%, respectively. Na2EDTA was recovered and recycled to the removal of heavy metals from MWIFA. Comparing new Na2EDTA solution with recycled Na2EDTA solution, recycled Na2EDTA and water could be reutilized to dispose MWIFA. The removal efficiencies of Cu, Pb, Zn, Cd, and Ni by recycled 0.2 mol/L Na2EDTA solution were 67.1%, 68.8%, 63.2%, 73.9%, 50.7%, respectively, the removal efficiencies using recovered Na2EDTA decreased by 2.6%, 3.9%, 3.3%, 4.2%, and 1.6%, respectively. Successive batch experiments were also conducted to evaluate industrialization potential and reusable times for recycled Na2EDTA. After four recirculation cycles, extraction efficiencies of Pb and Cd (removal efficiency at different cycles divided by removal efficiency of new Na2EDTA) declined toward 80%. Results from this research indicated that this circulating system possessed industrialization potential. Implications: An effective circulating system was developed to remove heavy metals in MWI fly ash (MWIFA). Integration of Na2EDTA with Fe0 promoted the removal of heavy metals from MWIFA. Na2EDTA, NaCl and water were stepwise extracted from iron-column effluent, respectively. Recovered Na2EDTA can still effectively remove heavy metals from MWIFA. Results from this research indicated that this circulating system possessed industrialization potential.

9.
Vet Parasitol ; 280: 109068, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32278937

RESUMO

Interleukin-4 (IL-4), an immunomodulatory cytokine derived from activated T lymphocytes were shown to regulate Th2-type immune responses. It plays an important role in anti-parasitic infections. In this study, a recombinant plasmid was designed using murine IL-4 co-expressed with pgsA anchor system of Lactobacillus plantarum NC8 and was delivered by live Lactobacillus plantarum NC8, which exhibited an enhanced immunogenicity in protection of BALB/c mice from infection with Trichinella spiralis. The results showed that the levels of serum IgG1 and mucosal secretory IgA (sIgA) were both increased significantly in mice orally inoculated with NC8-pSIP409-pgsA-mIL-4, and the Th2 phenotype immune response was up-regulated. A 29.9 % reduction in adult worm burden at 7 days post-infection (dpi) and 83.3 % reduction in muscle larvae burden at 28 dpi were observed in immune-stimulated mice with NC8-pSIP409-pgsA-mIL-4. Moreover, weight loss and pathological changes were also improved in mice of NC8-pSIP409-pgsA-mIL-4 group. Taken together, it suggests that NC8-pSIP409-pgsA-mIL-4 could improve the intestinal mucosal immunity and promoted the elimination of the adult worm in Trichinella-infected mice. This study laid the foundation for the development of a novel vaccines against Trichinellosis.


Assuntos
Interleucina-4/administração & dosagem , Lactobacillus plantarum/química , Probióticos/administração & dosagem , Trichinella spiralis/efeitos dos fármacos , Triquinelose/prevenção & controle , Administração Oral , Animais , Feminino , Interleucina-4/farmacologia , Lactobacillus plantarum/genética , Camundongos , Camundongos Endogâmicos BALB C , Microrganismos Geneticamente Modificados/química , Microrganismos Geneticamente Modificados/genética , Probióticos/química , Probióticos/farmacologia , Triquinelose/parasitologia
10.
PLoS Pathog ; 16(3): e1008429, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32208449

RESUMO

Chromatin dynamics regulated by epigenetic modification is crucial in genome stability and gene expression. Various epigenetic mechanisms have been identified in the pathogenesis of human diseases. Here, we examined the effects of ten epigenetic agents on pseudorabies virus (PRV) infection by using GFP-reporter assays. Inhibitors of bromodomain protein 4 (BRD4), which receives much more attention in cancer than viral infection, was found to exhibit substantial anti-viral activity against PRV as well as a range of DNA and RNA viruses. We further demonstrated that BRD4 inhibition boosted a robust innate immune response. BRD4 inhibition also de-compacted chromatin structure and induced the DNA damage response, thereby triggering the activation of cGAS-mediated innate immunity and increasing host resistance to viral infection both in vitro and in vivo. Mechanistically, the inhibitory effect of BRD4 inhibition on viral infection was mainly attributed to the attenuation of viral attachment. Our findings reveal a unique mechanism through which BRD4 inhibition restrains viral infection and points to its potent therapeutic value for viral infectious diseases.


Assuntos
Proteínas de Ciclo Celular/imunologia , Dano ao DNA/imunologia , Vírus de DNA/imunologia , Imunidade Inata , Proteínas Nucleares/imunologia , Vírus de RNA/imunologia , Fatores de Transcrição/imunologia , Células A549 , Animais , Chlorocebus aethiops , Infecções por Vírus de DNA/imunologia , Cães , Feminino , Células HEK293 , Células HeLa , Humanos , Células Madin Darby de Rim Canino , Camundongos , Camundongos Endogâmicos BALB C , Células NIH 3T3 , Células RAW 264.7 , Infecções por Vírus de RNA/imunologia , Suínos , Células Vero
11.
Parasit Vectors ; 13(1): 56, 2020 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-32046772

RESUMO

BACKGROUND: Eimeria spp. are responsible for chicken coccidiosis which is the most important enteric protozoan disease resulting in tremendous economic losses in the poultry industry. Understanding the interaction between the avian cecal microbiota and coccidia is of interest in the development of alternative treatments that do not rely on chemotherapeutics and do not lead to drug resistance. METHODS: We utilized 16S rRNA gene sequencing to detect the dynamics of the cecal microbial community in AA broilers challenged with Eimeria tenella. Histopathological analysis of the cecum was also conducted. RESULTS: We found that microbial shifts occur during the infection. Lactobacillus, Faecalibacterium, Ruminococcaceae UCG-013, Romboutsia and Shuttleworthia decreased in abundance. However, the opportunistic pathogens Enterococcus and Streptococcus increased in abundance over time in response to the infection. CONCLUSIONS: Eimeria tenella disrupts the integrity of the cecal microbiota and could promote the establishment and growth of potentially pathogenic bacteria. Defining bacterial populations affected by coccidial infection might help identify bacterial markers for intestinal disease as well as populations or species that could be beneficial in maintaining and restoring gut homeostasis during and after infection with E. tenella.


Assuntos
Galinhas , Coccidiose/veterinária , Eimeria tenella , Microbioma Gastrointestinal/genética , Doenças das Aves Domésticas/parasitologia , Animais , Bactérias/classificação , Bactérias/isolamento & purificação , Ceco/microbiologia , Ceco/parasitologia , Ceco/patologia , Galinhas/microbiologia , Galinhas/parasitologia , Coccidiose/terapia , Eimeria tenella/genética , Eimeria tenella/parasitologia , Metagenômica , RNA Ribossômico 16S/genética
12.
J Microbiol Biotechnol ; 30(4): 515-525, 2020 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-31838830

RESUMO

Interferon (IFN)-λ plays an essential role in mucosal cells which exhibit strong antiviral activity. Lactobacillus plantarum (L. plantarum) has substantial application potential in the food and medical industries because of its probiotic properties. Alphacoronaviruses, especially porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), cause high morbidity and mortality in piglets resulting in economic loss. Co-infection by these two viruses is becoming increasingly frequent. Therefore, it is particularly important to develop a new drug to prevent diarrhea infected with mixed viruses in piglets. In this study, we first constructed an anchored expression vector with CWA (C-terminal cell wall anchor) on L. plantarum. Second, we constructed two recombinant L. plantarum strains that anchored IFN-λ3 via pgsA (N-terminal transmembrane anchor) and CWA. Third, we demonstrated that both recombinant strains possess strong antiviral effects against coronavirus infection in the intestinal porcine epithelial cell line J2 (IPEC-J2). However, recombinant L. plantarum with the CWA anchor exhibited a more powerful antiviral effect than recombinant L. plantarum with pgsA. Consistent with this finding, Lb.plantarum-pSIP-409-IFN-λ3-CWA enhanced the expression levels of IFN-stimulated genes (ISGs) (ISG15, OASL, and Mx1) in IPEC-J2 cells more than did recombinant Lb.plantarum-pSIP-409-pgsA'-IFN-λ3. Our study verifies that recombinant L. plantarum inhibits PEDV and TGEV infection in IPEC-J2 cells, which may offer great potential for use as a novel oral antiviral agent in therapeutic applications for combating porcine epidemic diarrhea and transmissible gastroenteritis. This study is the first to show that recombinant L. plantarum suppresses PEDV and TGEV infection of IPEC-J2 cells.


Assuntos
Infecções por Coronavirus/veterinária , Gastroenterite Suína Transmissível/prevenção & controle , Interferons/administração & dosagem , Lactobacillus plantarum/genética , Doenças dos Suínos/prevenção & controle , Animais , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/virologia , Células Epiteliais/imunologia , Células Epiteliais/virologia , Feminino , Gastroenterite Suína Transmissível/genética , Gastroenterite Suína Transmissível/imunologia , Gastroenterite Suína Transmissível/virologia , Expressão Gênica , Interferons/genética , Interferons/imunologia , Lactobacillus plantarum/metabolismo , Masculino , Vírus da Diarreia Epidêmica Suína/fisiologia , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/imunologia , Doenças dos Suínos/virologia , Vírus da Gastroenterite Transmissível/fisiologia
13.
Int J Biol Macromol ; 151: 1181-1193, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-31743714

RESUMO

Interferon-inducible transmembrane proteins (IFITMs) restrict infection by several viruses, such as influenza A virus, West Nile virus and dengue virus. It has not been determined whether porcine IFITMs (pIFITMs) inhibit infection by pseudorabies virus (PRV), an enveloped, double-stranded DNA virus, which is the etiological agent of Aujeszky's disease in pigs. Here, we report that PRV infection elicited pIFITM1 expression in PK15 porcine kidney epithelial cells and 3D4/21 alveolar macrophages. pIFITM2 and pIFITM3 expression was only elevated in PK15 cells during PRV infection. Depletion of pIFITM1 using RNA interference, either in PK15 or in 3D4/21 cells, enhanced PRV infection while overexpression of pIFITM1 had the opposite effect. Knockdown of pIFITM2 and pIFITM3 did not influence PRV infection, suggesting that pIFITM2 and pIFITM3 are independent of PRV infection. PRV-induced pIFITM1 expression was dependent on the cGAS/STING/TBK1/IRF3 innate immune pathway and interferon-alpha receptor-1, suggesting that pIFITM1 is up-regulated by the type I interferon signaling pathway. The anti-PRV role of pIFITM1 was inhibited upon PRV entry. Our data demonstrate that pIFITM1 is a host restriction factor that inhibits PRV entry that may shed light on a strategy for prevention of PRV infection.

14.
Front Public Health ; 8: 587298, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33490017

RESUMO

Persistent infection with the carcinogenic human papillomavirus (HPV) is a prerequisite for the progression of cervical lesions and cancer. A growing body of research has focused on the functional role of the vaginal microbiota in the persistence of HPV infection. Understanding the microbial composition and structure in women with high-risk (hr)-HPV infection may help reveal associations between the vaginal microbiota and HPV infection, and identify potential biomarkers. Our study investigated the vaginal microbial community in women with and without hr-HPV infection, by using 16s rRNA gene sequencing. We found that microbial perturbations occurred in the early phase of hr-HPV infection. Lactobacillus and Sporolactobacillus were decreased, while bacteria related to bacterial vaginosis (BV), such as Gardnerella, Prevotella, Dialister, Slackia, Actinomyces, Porphyromonas, Peptoniphilus, Anaerococcus, Peptostreptococcus, Streptococcus, Ureaplasma, Megasphaera, and Mycoplasma were increased. Our results could offer insights into the correlations between hr-HPV and the vaginal microbiota in the early infection period, and provide indications that the predominance of some BV-associated bacteria during hr-HPV infection may increase the risk for cervical neoplasia.

15.
Front Microbiol ; 10: 2663, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31798571

RESUMO

Aeromonas veronii is an emerging aquatic pathogen causing hemorrhagic septicemia in humans and animals. Probiotic is an effective strategy for controlling enteric infections through reducing intestinal colonization by pathogens. Here we report that the consumption of Bacillus velezensis regulated the intestinal innate immune response and decreased the degree of intestinal inflammation damage caused by the A. veronii in Crucian carp. In this study, we isolated four strains of B. velezensis, named C-11, S-22, L-17 and S-14 from apparently healthy Crucian carp, which exerted a broad-spectrum antimicrobial activity inhibiting both Gram-positive and Gram-negative bacteria especially the fish pathogens. B. velezensis isolates showed typical Bacillus characteristics by endospore staining, physiological and biochemical test, enzyme activity analysis (amylase, protease, and lipase), and molecular identification. Here, Bacillus-containing dietary was orally administrated to Crucian carp for 8 weeks before A. veronii challenge. Immunological parameters and the expression of immune-related genes were measured at 2, 4, 6, 8, and 10 weeks post-administration. The results showed that B. velezensis was found to promote the increase in the phagocytic activities of peripheral blood leukocytes (PBLs) and head kidney leukocytes (HKLs), as well as the increase in interleukin 1ß (IL-1ß), IL-10 and tumor necrosis factor α (TNF-α) concentration of serum. Lysozyme levels (113.76 U/mL), ACP activity (25.32 U/mL), AKP activity (130.08 U/mL), and SOD activity (240.63 U/mL) were maximum (P < 0.05) in the B. velezensis C-11 treated group at 8 week. Our results showed that Crucian carp fed with the diet containing B. velezensis C-11 and S-22 developed a strong immune response with significantly higher (P < 0.05) levels of IgM in samples of serum, mucus of skin and intestine compared to B. velezensis L-17 and S-14 groups. Moreover, B. velezensis spores appeared to show no toxicity and damage in fish, which could inhabit the gut of Crucian carp. B. velezensis restrained up-regulation of pro-inflammation cytokines (IL-1ß, IFN-γ, and TNF-α) mRNA levels in the intestine and head kidney at final stage of administration, and the expression of IL-10 was increased throughout the 10-week trial. A. veronii infection increased the population of inflammatory cells in the intestinal villi in the controls. In contrast, numerous goblet cells and few inflammatory cells infiltrated the mucosa in the B. velezensis groups after challenge with A. veronii. Compared with A. veronii group, B. velezensis could safeguard the integrity of intestinal villi. The highest post-challenge survival rate (75.0%) was recorded in B. velezensis C-11 group. The present data suggest that probiotic B. velezensis act as a potential gut-targeted therapy regimens to protecting fish from pathogenic bacteria infection. IMPORTANCE: In this work, four Bacillus velezensis strains isolated from apparently healthy Crucian carp, which exhibited a broad-spectrum antibacterial activity especially the fish pathogens. Administration of B. velezensis induced the enhancement of the intestinal innate immune response through reducing intestinal colonization by pathogens. The isolation and characterization would help better understand probiotic can be recognized as an alternative of antimicrobial drugs protecting human and animal health.

16.
World J Gastroenterol ; 25(24): 3044-3055, 2019 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-31293340

RESUMO

BACKGROUND: The formation of liver fibrosis is mainly caused by the activation of hepatic stellate cells (HSCs) and the imbalance of extracellular matrix (ECM) production and degradation. The treatment of liver fibrosis mainly includes removing the cause, inhibiting the activation of HSCs, and inhibiting inflammation. NOD-like receptor (NLR) family, caspase activation and recruitment domain (CARD) domain containing 5/NOD27/CLR16.1 (NLRC5) is a highly conserved member of the NLR family and is involved in inflammation and immune responses by regulating various signaling pathways such as nuclear factor-κB (NF-κB) signaling. It has been found that NLRC5 plays an important role in liver fibrosis, but its specific effect and possible mechanism remain to be fully elucidated. AIM: To investigate the role of NLRC5 in the activation and reversion of HSCs induced with transforming growth factor-ß (TGF-ß) and MDI, and to explore its relationship with liver fibrosis. METHODS: A total of 24 male C57BL/6 mice were randomly divided into three groups, including normal, fibrosis, and recovery groups. Twenty-four hours after a liver fibrosis and spontaneous reversion model was established, the mice were sacrificed and pathological examination of liver tissue was performed to observe the degree of liver fibrosis in each group. LX-2 cells were cultured in vitro and treated with TGF-ß1 and MDI. Real-time quantitative PCR (qPCR) and Western blot were used to analyze the expression levels of NLRC5, α-smooth muscle actin (α-SMA), and collagen type I alpha1 (Col1a1) in each group. The activity of NF-κB in each group of cells transfected with NLRC5-siRNA was detected. RESULTS: Compared with the normal mice, the expression level of NLRC5 increased significantly (P < 0.01) in the fibrosis group, but decreased significantly in the recovery group (P < 0.01). In in vitro experiments, the content of NLRC5 was enhanced after TGF-ß1 stimulation and decreased to a lower level when treated with MDI (P < 0.01). The expression of α-SMA and Col1a1 proteins and mRNAs in TGF-ß1-mediated cells was suppressed by transfection with NLRC5-siRNA (P < 0.01). Western blot analysis showed that the expression of NF-κB p65 protein and phosphorylated IκBα (p-IκBα) was increased in the liver of mice in the fibrosis group but decreased in the recovery group (P < 0.01), and the protein level of nuclear p65 and p-IκBα was significantly increased after treatment with NLRC5-siRNA (P < 0.01). CONCLUSION: NLRC5 may play a key role in the development and reversal of hepatic fibrosis through the NF-κB signaling pathway, and it is expected to be one of the clinical therapeutic targets.


Assuntos
Células Estreladas do Fígado/patologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Cirrose Hepática Experimental/patologia , NF-kappa B/metabolismo , Animais , Tetracloreto de Carbono/toxicidade , Linhagem Celular , Matriz Extracelular/patologia , Técnicas de Silenciamento de Genes , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Fígado/citologia , Fígado/patologia , Cirrose Hepática Experimental/induzido quimicamente , Masculino , Camundongos , RNA Interferente Pequeno/metabolismo , Proteínas Recombinantes/metabolismo , Transdução de Sinais , Fator de Crescimento Transformador beta1/metabolismo
17.
J Nanosci Nanotechnol ; 19(7): 4058-4063, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30764970

RESUMO

A facile one-pot hydrothermal method for fabricating nitrogen-doped carbon dots (N-CDs) was developed by using citric acid as a carbon source and guanidine carbonate as a nitrogen and carbon source. X-ray diffraction, X-ray photoelectron spectroscopy, and Fourier transform infrared spectra indicated that the N-CDs were rich in elemental nitrogen. They had excellent stability in the presence of various salt concentrations and under UV irradiation. The N-CDs exhibited high quantum yields (52%), as well as down-conversion and up-conversion photoluminescence. The N-CD photoluminescence was quenched in the presence of Hg2+, while nearly no intensity changes were observed when in the presence of Na+, Mg2+, Mn2+, Zn2+, Ni2+, Cu2+, Ba2+, Cd2+ or Ca2+. The binding constant (KSV) and detection limit were also determined.

18.
Virus Res ; 263: 64-72, 2019 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-30611823

RESUMO

To evaluate the efficiency of preventing pathogenic avian influenza by vaccination with recombinant Lactobacillus plantarum (L. plantarum) that expresses conserved antigens, the mucosal and systemic immune responses in animals vaccinated with recombinant L. plantarum NC8-409-1 (NC8-pSIP409-pgsA'-HA2) and NC8-409-2 (NC8-pSIP409-pgsA'-3M2e-HA2) were evaluated. Our results showed that recombinant L. plantarum NC8-409-1 and NC8-409-2 could substantially stimulate the specific IgA titer in the intestine and the specific IgG antibody titer in the serum. We also found that recombinant L. plantarum induced increases in the number of B220+ IgA+ cells in Peyer's patches (PPs), in lymphocyte proliferation and in the number of IFN-γ+ producing CD8+ T cells after immunization in mice. Most importantly, the mice that were vaccinated with recombinant L. plantarum NC8-409-2 and NC8-409-1 were to some extent protected against infection challenge with the H9N2 and H1N1 viruses. In particular, NC8-409-2 provided up to 80% protection against the H9N2 virus, and NC8-409-1 provided up to 60% protection. Lung tissue pathology was also reduced. Therefore, recombinant L. plantarum NC8-409-2 and NC8-409-1 may be candidate oral vaccines against bird flu.


Assuntos
Portadores de Fármacos , Hemaglutininas Virais/imunologia , Vacinas contra Influenza/imunologia , Lactobacillus plantarum/genética , Infecções por Orthomyxoviridae/prevenção & controle , Proteínas Recombinantes de Fusão/imunologia , Proteínas da Matriz Viral/imunologia , Animais , Anticorpos Antivirais/análise , Anticorpos Antivirais/sangue , Sangue/imunologia , Linfócitos T CD8-Positivos/imunologia , Proliferação de Células , Modelos Animais de Doenças , Hemaglutininas Virais/genética , Imunoglobulina A/análise , Imunoglobulina G/sangue , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H9N2/imunologia , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Interferon gama/metabolismo , Intestinos/imunologia , Pulmão/patologia , Camundongos , Proteínas Recombinantes de Fusão/genética , Análise de Sobrevida , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Proteínas da Matriz Viral/genética
19.
Inorg Chem ; 58(2): 1309-1316, 2019 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-30620578

RESUMO

Six analogue compounds with the general formula [Fe2( xL)5(NCS)4]· yMeOH ( x = o-Cl, y = 3 for compound 1; x = m-Cl, y = 5 for 2; x = p-Cl, y = 1 for 3; x = o-Me, y = 2 for 4; x = m-Me, y = 2 for 5; x = p-Me, y = 3 for 6; L = N-phenylmethylene-4-amino-1,2,4-triazole) were synthesized. The two Fe(II) ions are triply bridged by the triazole groups of three xL ligands and each Fe(II) is further capped with two NCS- groups and one more xL ligand. These compounds show regular patterns in their magnetic properties that depend on the positions the substituent groups (-Cl or -Me) ride, i.e., ortho-substituted compounds 1 and 4 undergo complete one-step spin crossover (SCO), while meta-substituted compounds 2 and 5 display incomplete one-step SCO with lower transition temperatures, and para-substituted compounds 3 and 6 are in the high-spin states in all temperature ranges. Structural analyses reveal that the molecular geometry and intermolecular interactions of these compounds, which should account for the differences in magnetic properties, are obviously depend on the positions of substituent groups (steric effect), despite them being electron-withdrawing chlorine or electron-donating methyl, whereas theoretical calculations confirm that the electronic effects of substituent groups exert no effect on the magnetic properties.

20.
Parasitology ; 146(4): 521-526, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30427300

RESUMO

The present study was undertaken to investigate the antiparasitic activity of extracellular products of Streptomyces albus. Bioactivity-guided isolation of chloroform extracts affording a compound showing potent activity. The structure of the compound was elucidated as salinomycin (SAL) by EI-MS, 1H NMR and 13C NMR. In vitro test showed that SAL has potent anti-parasitic efficacy against theronts of Ichthyophthirius multifiliis with 10 min, 1, 2, 3 and 4 h (effective concentration) EC50 (95% confidence intervals) of 2.12 (2.22-2.02), 1.93 (1.98-1.88), 1.42 (1.47-1.37), 1.35 (1.41-1.31) and 1.11 (1.21-1.01) mg L-1. In vitro antiparasitic assays revealed that SAL could be 100% effective against I. multifiliis encysted tomonts at a concentration of 8.0 mg L-1. In vivo test demonstrated that the number of I. multifiliis trophonts on Erythroculter ilishaeformis treated with SAL was markedly lower than that of control group at 10 days after exposed to theronts (P < 0.05). In the control group, 80% mortality was observed owing to heavy I. multifiliis infection at 10 days. On the other hand, only 30.0% mortality was recorded in the group treated with 8.0 mg L-1 SAL. The median lethal dose (LD50) of SAL for E. ilishaeformis was 32.9 mg L-1.


Assuntos
Antiprotozoários/farmacologia , Infecções por Cilióforos/veterinária , Cyprinidae , Doenças dos Peixes/tratamento farmacológico , Hymenostomatida/efeitos dos fármacos , Piranos/farmacologia , Streptomyces/química , Animais , Infecções por Cilióforos/tratamento farmacológico
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