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1.
J Agric Food Chem ; 71(22): 8400-8412, 2023 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-37246803

RESUMO

The oriental fruit fly, Bactrocera dorsalis, is a damaging insect pest for many vegetable and fruit crops that has evolved severe chemical insecticide resistance, including organophosphorus, neonicotinoid, pyrethroid, and macrolides. Hence, it is important to elucidate its detoxification mechanism to improve its management and mitigate resource destruction. Glutathione S-transferase (GST) is a critical secondary phase enzyme that plays multiple detoxification functions against xenobiotics. In this study, we identified several BdGSTs by characterizing their potential relationships with five insecticides using inducible and tissue-specific expression pattern analyses. We found that an antenna-abundant BdGSTd8 responded to four different classes of insecticides. Subsequently, our immunohistochemical and immunogold staining analysis further confirmed that BdGSTd8 was primarily located in the antenna. Our investigations also confirmed that BdGSTd8 possesses the capability to enhance cell viability by directly interacting with malathion and chlorpyrifos, which clarified the function of antenna-abundant GST in B. dorsalis. Altogether, these findings enrich our understanding of GST molecular characteristics in B. dorsalis and provide new insights into the detoxification of superfluous xenobiotics in the insect antenna.


Assuntos
Inseticidas , Tephritidae , Animais , Inseticidas/farmacologia , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Xenobióticos , Compostos Organofosforados , Tephritidae/genética , Tephritidae/metabolismo
2.
Huan Jing Ke Xue ; 44(3): 1440-1451, 2023 Mar 08.
Artigo em Chinês | MEDLINE | ID: mdl-36922205

RESUMO

To analyze spatial-temporal variation in groundwater quality and contamination sources in the Shihezi-Changji area in Xinjiang, a Bayesian water quality evaluation model based on Shannon entropy, Spearman rank correlation coefficient, principal component analysis, and an absolute principal component scores-multiple linear regression receptor model (APCS-MLR) were comprehensively used in this study. Groundwater samples in 23 in-situ wells were collected from 2016 to 2021 for quality analysis. The results showed that ① groundwater quality was generally good, with most samples having a phreatic water quality of Class Ⅰ and Class Ⅱ and most confined groundwater quality being of Class Ⅰ. ② Temporally, 2016 and 2017 were the key time nodes of water quality variation in phreatic water and confined groundwater, respectively. Class Ⅳ and Class Ⅴ groundwater was observed only before the key time nodes, whereas after those time nodes the groundwater quality fluctuated within Class Ⅰ to Class Ⅲ. ③ Spatially, the order of phreatic water quality in descending order was Shihezi City, Hutubi county, Manas county, and Changji City, whereas that of confined groundwater quality was:Shihezi City and Changji City, Hutubi county, and Manas county. ④ The spatial-temporal variations in groundwater quality and that in major related indices were basically similar and highly heterogeneous. ⑤ Phreatic water quality was mainly affected by leaching (67.30%), leaching-migration (10.89%), and agricultural-domestic pollution (9.44%); by contrast, unconfined groundwater quality was mainly affected by leaching-enrichment (52.08%), agricultural-domestic pollution (16.06%), and ion exchange under an alkaline environment (12.64%). Although groundwater quality was improved over the years, more attention should be paid to groundwater salinization in the 149 Regiment in northern Manas County.

3.
Biochem Biophys Res Commun ; 645: 55-60, 2023 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-36680937

RESUMO

Chronic pain is frequently reported in clinical practice. Therefore, it is important to identify effective therapy to relieve pain. In this work, we selected Forsythoside B (FB), a phenylethanoid glycoside isolated from Forsythia suspensa (Thunb.) Vahl, to evaluate its effect in modulating inflammatory pain induced by complete Freund's adjuvant (CFA) and the involved mechanisms. We discovered that FB could attenuate inflammatory pain triggered by CFA injection and exert anti-anxiety effects. In detail, proinflammatory cytokines, consisting of IL-6 and TNF-α, were decreased after FB administration in the CFA-injected mice. Furthermore, the FB application ameliorated the activation of ionized calcium-binding adaptor molecule 1 (Iba-1) and glial fibrillary acidic protein (GFAP), the microglia and astrocytes markers respectively. Therefore, our findings indicate that FB could be a promising treatment for chronic inflammatory pain.


Assuntos
Dor Crônica , Inflamação , Camundongos , Animais , Adjuvante de Freund , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Anti-Inflamatórios/uso terapêutico , Anti-Inflamatórios/farmacologia , Dor Crônica/induzido quimicamente , Dor Crônica/tratamento farmacológico , Glucosídeos/farmacologia , Glucosídeos/uso terapêutico , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Hiperalgesia/metabolismo
4.
Pest Manag Sci ; 79(2): 666-677, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36223172

RESUMO

BACKGROUND: The oriental fruit fly, Bactrocera dorsalis (Hendel) is a worldwide pest damaging a wide range of hosts. Due to the long-term indiscriminate use of insecticides, B. dorsalis has developed serious resistance to several insecticides. UDP-glycosyltransferases (UGTs) are secondary metabolic enzymes involved in biotransformation and play an important role in the metabolism of plant secondary metabolites and synthetic insecticides in insects. Thus, we suspect that UGTs in B. dorsalis play an important role in insecticide tolerance. RESULTS: In this study, 31 UGT genes were identified in the genome of B. dorsalis, belonging to 13 subfamilies. Real-time quantitative polymerase chain reaction (RT-qPCR) results revealed that 12 UGT genes were highly expressed in the antennae, midgut, Malpighian tubule and fat body. The mRNA expressions of 17 UGT genes were up-regulated upon exposure to λ-cyhalothrin, imidacloprid, abamectin and chlorpyrifos. Knockdown of the selected five UGT genes (BdUGT301D2, BdUGT35F2, BdUGT36K2, BdUGT49D2, BdUGT50B5) by RNA interference increased the mortality of B. dorsalis from 9.29% to 27.22% upon exposure to four insecticides. CONCLUSION: The abundance of UGTs in B. dorsalis is similar to other insect species, and 12 out of 31 UGTs were specifically expressed in metabolic tissues, suggesting a key role in detoxification. Down-regulation of five selected UGT genes increased the susceptibility of B. dorsalis to various insecticides, indicating that UGTs may play an important role in tolerance of B. dorsalis to multiple insecticides. © 2022 Society of Chemical Industry.


Assuntos
Inseticidas , Tephritidae , Animais , Inseticidas/farmacologia , Difosfato de Uridina , Insetos/metabolismo , Drosophila , Glicosiltransferases/genética
5.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 30(6): 1757-1765, 2022 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-36476900

RESUMO

OBJECTIVE: To explore the correlation between different body mass index (BMI) and prognosis of mantle cell lymphoma (MCL). METHODS: The clinical characteristics and biological indices of 108 patients with MCL treated in Fujian Medical University Union Hospital were retrospectively analyzed, and the effects of different BMI on overall survival (OS) and progression-free survival (PFS) were analyzed. The correlation between BMI and B symptoms, LDH and Ki-67 was further observed. Furthermore,the differences of BMI between Autologous peripheral blood stem cell transplantation(Auto-PBSCT) and conventional chemotherapy groups were explored. RESULTS: Among 108 patients, the median age at diagnosis was 59(25-79) years old, and the male to female ratio was 4.4∶1. 88.89% of patients with Ann Arbor staging III-IV, 63.89% with bone marrow involvement, and 49.07% with splenic infiltration. Patients with BMI ≥ 24 kg/m2 were divided into two groups: the high BMI group and the low BMI group. The 5-year PFS and OS of patients in the low BMI group were 31.9% and 47.0%, respectively, while those in the high BMI group were 64.6% and 68.7%, respectively. The incidence of death in the high BMI group was lower than that of the low BMI group (P<0.01). In multivariate analysis, BMI was an independent predictor of PFS (HR=0.282; 95% CI: 0.122-0.651; P=0.003) and an independent predictor of OS (HR=0.299; 95% CI: 0.129-0.693; P=0.005). Also, patients with B symptoms had a lower BMI than those without B symptoms (P=0.01), but BMI had no effect on patients' LDH and Ki-67. The prognosis of 16 patients treated with Auto-PBSCT was significantly better than that of the conventional chemotherapy group. There was no significant difference in BMI between Auto-PBSCT group and conventional chemotherapy group. CONCLUSION: BMI is an independent prognostic factor for PFS and OS in MCL, and may be influenced by the effect of B symptoms on BMI.


Assuntos
Linfoma de Célula do Manto , Adulto , Humanos , Feminino , Masculino , Pessoa de Meia-Idade , Idoso , Linfoma de Célula do Manto/terapia , Índice de Massa Corporal , Antígeno Ki-67 , Estudos Retrospectivos , Prognóstico
6.
Acta Pharmacol Sin ; 43(11): 2828-2840, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35577909

RESUMO

Sevoflurane inhalation is prone to initiate cognitive deficits in infants. The early growth response-2 (Egr-2) gene is DNA-binding transcription factor, involving in cognitive function. In this study we explored the molecular mechanisms underlying the vulnerability to cognitive deficits after sevoflurane administration. Six-day-old (young) and 6-week-old (early adult) mice received anesthesia with 3% sevoflurane for 2 h daily for 3 days. We showed that multiple exposures of sevoflurane induced significant learning ability impairment in young but not early adult mice, assessed in Morris water maze test on postnatal days 65. The integrated differential expression analysis revealed distinct transcription responses of Egr family members in the hippocampus of the young and early adult mice after sevoflurane administration. Particularly, Egr2 was significantly upregulated after sevoflurane exposure only in young mice. Microinjection of Egr2 shRNA recombinant adeno-associated virus into the dentate gyrus alleviated sevoflurane-induced cognitive deficits, and abolished sevoflurane-induced dendritic spins loss and BDNF downregulation in young mice. On the contrary, microinjection of the Egr2 overexpression virus in the dentate gyrus aggravated learning ability impairment induced by sevoflurane in young mice but not early adult mice. Furthermore, we revealed that sevoflurane markedly upregulated the nuclear factors of activated T-cells NFATC1 and NFATC2 in young mice, which were involved in Egr2 regulation. In conclusion, Egr2 serves as a critical factor for age-dependent vulnerability to sevoflurane-induced cognitive deficits.


Assuntos
Anestésicos Inalatórios , Disfunção Cognitiva , Proteína 2 de Resposta de Crescimento Precoce , Éteres Metílicos , Animais , Camundongos , Anestésicos Inalatórios/toxicidade , Animais Recém-Nascidos , Cognição , Disfunção Cognitiva/induzido quimicamente , Proteína 2 de Resposta de Crescimento Precoce/genética , Proteína 2 de Resposta de Crescimento Precoce/metabolismo , Hipocampo/metabolismo , Aprendizagem em Labirinto , Sevoflurano/efeitos adversos
7.
Pharm Biol ; 60(1): 195-205, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35060427

RESUMO

CONTEXT: Folium Ginkgo extract and tetramethylpyrazine sodium chloride injection (Xingxiong injection) is a compound preparation commonly used for treating cerebral ischaemia/reperfusion injury in ischaemic stroke in China. However, its potential mechanisms on ischaemic stroke remain unknown. OBJECTIVE: This study explores the potential mechanisms of Xingxiong injection in vivo or in vitro. MATERIALS AND METHODS: Sprague-Dawley (SD) rats were randomly assigned to five groups: the sham (normal saline), the model (normal saline) and the Xingxiong injection groups (12.5, 25 or 50 mL/kg). The rats were subjected to 2 h of middle cerebral artery occlusion (MCAO) followed by reperfusion for 14 d. Xingxiong injection was administered via intraperitoneal (i.p.) injection immediately after ischaemia induction for 14 d. Afterwards, rats were sacrificed at 14 d induced by administration of Xingxiong injection. RESULTS: Xingxiong injection significantly reduces infarct volume (23%) and neurological deficit scores (93%) compared with the MCAO/R group. Additionally, Xingxiong injection inhibits the loss in mitochondrial membrane potential (43%) and reduces caspase-3 level (44%), decreases NOX (41%), protein carbonyl (29%), 4-HNE (40%) and 8-OhdG (41%) levels, inhibits the expression of inflammatory factors, such as TNF-α (26%), IL-1ß (34%), IL-6 (39%), MCP-1 (36%), CD11a (41%) and ICAM-1 (43%). Moreover, Xingxiong injection can increase p-Akt/Akt (35%) and Nrf2 (47%) protein expression and inhibit NLRP3 (42%) protein expression. CONCLUSIONS: Xingxiong injection prevents cerebral ischaemia/reperfusion injury via activating the Akt/Nrf2 pathway and inhibiting NLRP3 inflammasome. These findings provide experimental evidence for clinical use of drugs in the treatment of ischaemic stroke.


Assuntos
Isquemia Encefálica/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Ginkgo biloba/química , Traumatismo por Reperfusão/tratamento farmacológico , Animais , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/química , Infarto da Artéria Cerebral Média , Inflamassomos/metabolismo , AVC Isquêmico/tratamento farmacológico , Masculino , Fator 2 Relacionado a NF-E2/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Pirazinas/química , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley
8.
Virology ; 549: 77-84, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32853849

RESUMO

Human infections with highly pathogenic avian influenza (HPAI) H7N9 virus were detected in late 2016. We examined the drug resistance profile of 30 HPAI H7N9 isolates from Mainland of China (2016-2019). Altogether, 23% (7/30) carried neuraminidase inhibitors (NAIs) - resistance mutations, and 13% (4/30) displayed reduced susceptibility to NAIs in neuraminidase (NA) inhibition test. An HPAI H7N9 reassortment virus we prepared was passaged with NAIs for 10 passages. Passage with zanamivir induced an E119G substitution in NA, whereas passage with oseltamivir induced R292K and E119V substitutions that simulated that seen in oseltamivir -treated HPAI H7N9 cases, indicating that the high frequency of resistant strains in the HPAI H7N9 isolates is related to NAIs use. In presence of NAIs, R238I, A146E, G151E and G234T substitutions were found in HA1 region of HA. No amino acid mutations were found in the internal genes of the recombinant virus.


Assuntos
Farmacorresistência Viral/genética , Subtipo H7N9 do Vírus da Influenza A/genética , Mutação , Neuraminidase/genética , Vírus Reordenados/genética , Proteínas Virais/genética , Substituição de Aminoácidos , Animais , Antivirais/farmacologia , Aves/virologia , Inibidores Enzimáticos/farmacologia , Expressão Gênica , Glicoproteínas de Hemaglutininação de Vírus da Influenza/química , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Humanos , Subtipo H7N9 do Vírus da Influenza A/efeitos dos fármacos , Subtipo H7N9 do Vírus da Influenza A/metabolismo , Subtipo H7N9 do Vírus da Influenza A/patogenicidade , Influenza Aviária/patologia , Influenza Aviária/transmissão , Influenza Aviária/virologia , Influenza Humana/patologia , Influenza Humana/transmissão , Influenza Humana/virologia , Testes de Sensibilidade Microbiana , Modelos Moleculares , Neuraminidase/metabolismo , Oseltamivir/farmacologia , Conformação Proteica , Vírus Reordenados/efeitos dos fármacos , Vírus Reordenados/metabolismo , Vírus Reordenados/patogenicidade , Proteínas Virais/metabolismo , Zanamivir/farmacologia
9.
Infect Dis Poverty ; 7(1): 117, 2018 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-30514379

RESUMO

BACKGROUND: The injection of rabies immune globulin (RIG) is of the utmost importance in the management of category III exposures to rabies-suspect animals. Because of the high cost and limited availability of existing RIG, one possible replacement for RIG is monoclonal antibodies (MAbs) against the rabies virus (RABV). Consequently, it is necessary to determine the neutralizing activity of the MAbs against rabies viruses, especially street rabies virus. However, the method to detect the neutralizing activity of MAbs against street rabies virus remains undefined. METHODS: To establish a method for detecting the neutralizing activity of MAbs against street rabies virus, we constructed a library consisting of 12 strains of street RABV from 11 provinces in China. Using this street RABV library and the Reed-Muench formula, we established a method for detecting the neutralizing titer of the MAbs. The reliability and repeatability of the method were evaluated by repeatedly measuring the neutralizing activity of a MAb and a post vaccination serum. RESULTS: A total of 12 strains of street RABV were chosen for inclusion in the street RABV library, which covered six Chinese lineages (China I-China VI) and grew to high titers in N2A cells (> 105 FFD50/ml). On the basis of the library, we constructed the method to detect the neutralizing activity of the MAbs. The results of repeatedly measuring the MAbs and positive serum showed excellent reliability and repeatability of the method established in this study. CONCLUSIONS: This study established a street RABV library reflecting the epidemiological features of Chinese rabies viruses, which provides a platform for detecting the neutralizing activity of MAbs against rabies viruses circulating in China.


Assuntos
Vírus da Raiva/genética , Raiva/prevenção & controle , Animais , Anticorpos Monoclonais , Anticorpos Antivirais/imunologia , China/epidemiologia , Biblioteca Gênica , Humanos , Imunoglobulinas/uso terapêutico , Testes de Neutralização , Filogenia , Raiva/epidemiologia , Vacina Antirrábica , Vírus da Raiva/classificação , Vírus da Raiva/imunologia
10.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 26(4): 984-992, 2018 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-30111395

RESUMO

OBJECTIVE: To investigate the expression and clinical significances of HGFA, Matriptase, HAI-1 and HAI-2 in patients with acute myeloid leukemia (AML). METHODS: The bone marrow samples from 91 AML patients, 41 AML patients in complete remission, and 32 normal controls were collected. Real time fluorescence quantitative RT-PCR (qRT-PCR) was used to detect the mRNA expressions levels of HGFA, Matriptase, HAI-1, HAI-2 . The expressions of these genes were compared among AML untreated group, the complete remission group and the healthy control group. The correlation of their expression with clinical characteristics was analyzed. RESULTS: The level of HGFA in the AML untreated group was higher than that in the healthy control group(P<0.05), while the HAI-2 mRNA level was lower than that in the healthy control group(P<0.05). The mRNA levels of HAI-1 and Matriptase were not changed significantly in all groups. The HAI-2 mRNA expression level was significantly lower in the high white blood cell group (P<0.05). CONCLUSION: The abnormal activation of HGF/c-Met signaling system in AML may result from the increase of HGFA expression and the decrease of HAI-2 expression of the upstream regulatory factors.


Assuntos
Leucemia Mieloide Aguda , Fator de Crescimento de Hepatócito , Humanos , Glicoproteínas de Membrana , Proteínas Secretadas Inibidoras de Proteinases , Serina Endopeptidases
11.
Int J Mol Sci ; 19(5)2018 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-29693643

RESUMO

To investigate the roles and explore the altered expression of microRNAs (miRNAs) and mRNAs in chicken embryos in response to Newcastle disease virus (NDV) infection, deep sequencing was performed. Then, a conjoint analysis of small RNA-seq and mRNA-seq was performed to screen interactional miRNA⁻mRNA pairs during NDV infection. In total, 15 and 17 up- and downregulated miRNAs were identified that potentially targeted 4279 and 6080 mRNAs in NDV-infected chicken embryonic tissues, respectively; in addition, 595 upregulated and 480 downregulated mRNAs were identified. The conjoint analysis of the obtained data identified 1069 miRNA⁻mRNA pairs. Among these pairs, 130 pairs were related to immune or inflammatory responses. The relationship between gga-miR-203a and its target transglutaminase 2 (TGM2) was confirmed using a dual-luciferase reporter system and a real time quantitative polymerase chain reaction (RT-qPCR) assay. Overall, the discovery of miRNAs, mRNAs, and their potential pairing relationships, which may be involved in the regulation of NDV infection, will facilitate our understanding of the complex regulatory relationship between the host and the virus.


Assuntos
MicroRNAs/genética , Doença de Newcastle/genética , RNA Mensageiro/genética , Vísceras/metabolismo , Animais , Embrião de Galinha , Proteínas de Ligação ao GTP/genética , Proteínas de Ligação ao GTP/metabolismo , Células HEK293 , Humanos , MicroRNAs/metabolismo , Doença de Newcastle/metabolismo , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/patogenicidade , Proteína 2 Glutamina gama-Glutamiltransferase , RNA Mensageiro/metabolismo , Transcriptoma , Transglutaminases/genética , Transglutaminases/metabolismo , Vísceras/virologia
12.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 23(3): 689-92, 2015 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-26117018

RESUMO

OBJECTIVE: To explore the molecular pathological diagnosis of mucosa-associated lymphoid tissue lymphoma(MALTL). METHODS: Sixty MALTL paraffin embedding specimens were analyzed retrospectively. The distribution of the primary lesion, morphology, immunophenotype, IgH gene cloning rearrangement were evaluated. RESULTS: The main risk area for the patients with MALTL was gastric area(37%), in the second place was salivary gland(20%), in the third place was intestine (12%), orbit and ocular adnexa(12%); at low magnification, MALTL specimens manifasted diffuse growth majority, a few of nodular structure, lymphoma cell forms were diversified; The results of immunohistochemical detection showed that the CD20 and the BCL-2 were positive, the CD3, CD5, CD10, CD23, cyclin D1 and CD21 were negative, 17 specimen kappa or lambda express more obviously, their sensibility was 28.33%(17/60); 61.67%(37/60) developed IgH gene rearrangement, 19 specimen IgH gene rearrangement monoclonal and kappa or lambda were negative, the positive rate of both combined detections was 68.33%. CONCLUSION: The tissue morphologic characteristics and immuno-histochemistry detection are the basic means for MALTL diagnosis, the detection of IgH gene reasragement and Kappa or lamda restrictive expression has the practical importance for MALTL diagnosis, both combination can show higher positive rate for MALTL diagnosis.


Assuntos
Linfoma de Zona Marginal Tipo Células B , Ciclina D1 , Rearranjo Gênico , Humanos , Imunofenotipagem
13.
Yi Chuan ; 35(12): 1377-83, 2013 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-24645347

RESUMO

MNSFbeta (Monoclonal nonspecific suppressor factor beta) is a natural immunosuppressive factor which has been reported to be involved in various biological processes, such as immune responses, cell division, stress response, cell apoptosis, and nuclear transport. However, study on porcine MNSFbeta has been rarely reported. In this study, the full-length sequence of porcine MNSFbeta (GenBank accession number: KF77642500) was predicted in silicon and its cDNA sequence was obtained through RT-PCR from porcine spleen. The nucleic acid and protein sequences were analyzed. Then, the gene was subcloned into pEGFP-C1 to construct a recombinant plasmid pEGFP-MNSFbeta which was transfected into swine umbilical vein endothelial cells (SUVECs) using Lipofectamine 2000. The expression of GFP was detected by fluorescence microscopy, Western blot, and laser confocal fluorescence microscopy. The spatial expression patterns of porcine MNSFbeta were detected by real-time qPCR. Results showed that the full length of porcine MNSFbeta was 402 bp encoding 133 amino acids with only one exon. Bioinformatics analysis showed that porcine MNSFbeta protein was a stable protein consisting of a ubiquitin-like domain fused to the ribosomal protein S30 with no signal peptide. The analyses of homology and phylogenetic tree of porcine MNSFbeta and its homologs in other 18 species showed that the identities of MNSFbeta protein sequence were higher than 91% among different species and the evolutionary distance was less than 0.05. It indicates that MNSFbeta is highly conserved in the process of evolution. Fluorescence signal showed that the fusion protein GFP-MNSFbeta was successfully expressed in SUVECs which was then confirmed by Western blot. Laser confocal fluorescence microscopy showed that MNSFbeta was expressed in both nucleus and cytoplasm. Analysis of spatial expression patterns showed that procine MNSFbeta was widely expressed in immune tissues, but not in lung, suggesting that MNSFbeta may play an important role in immune response.


Assuntos
Fatores Supressores Imunológicos/metabolismo , Animais , Western Blotting , Biologia Computacional , Masculino , Estrutura Secundária de Proteína , Fatores Supressores Imunológicos/química , Fatores Supressores Imunológicos/genética , Suínos
14.
Zhonghua Liu Xing Bing Xue Za Zhi ; 33(3): 328-31, 2012 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-22613389

RESUMO

OBJECTIVE: To develop a singleplex PCR assay targeting O-antigen modification genes for molecular serotyping of Shigella (S.) flexneri. METHODS: Eight pairs of primer for O-antigen synthesis and modification genes of S. flexneri were designed and used for developing an O-antigen modification gene-specific singleplex PCR assay to serotype 14 most common S. flexneri serotypes (1a, 1b, 1c, 2a, 2b, 3a, 3b, 4a, 4b, 5a, Y, X, Xv and F6). Bacterial pathogens which causing diarrheal disease were used for specificity detection. 106 S. flexneri clinical isolates were serotyped by this method and compared with the slide agglutination method. RESULTS: An O-antigen modification, gene-specific singleplex PCR was developed. When six singleplex PCR reactions were performed, 14 of the 15 recognized S. flexneri serotypes were identified, except for serotype Xv. The detection threshold ranged from 10 pg to 1 ng DNA in a 20 µl reaction system. A high concordance between the singleplex PCR assay and slide agglutination were observed when 106 S. flexneri strains of various serotypes were analyzed with an exception that 1 serotype Y strain showed that it was carrying the additional defective gtr II genes. CONCLUSION: This method showed advantages over the traditional slide agglutination methods, and was promising when under application in the following situations as clinical diagnosis.


Assuntos
Reação em Cadeia da Polimerase/métodos , Sorotipagem , Shigella flexneri/classificação , Antígenos O/genética
15.
Int J Biol Macromol ; 50(3): 713-9, 2012 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-22214825

RESUMO

The objective of the current study is to investigate the protective effect of carboxymethylpachymaran (CMP) on the immune system of mice via multiple infections with porcine circovirus type 2 (PCV2). The in vivo results showed that CMP administration significantly improved the spleen or thymus index, promoted the proliferation activities of T or B lymphocytes, and increased the production of glutathione, the superoxidase dismutase capacity, and the total antioxidant capacity in the spleen or thymus of PCV2-infected mice. The administration of different CMP doses to PVC2-inoculated mice resulted in the upregulation of IL-2 and IFN-α or the downregulation of IL-10 levels in the serum. These findings suggest that CMP has potential applications in regulating immunological functions to overcome the immunosuppresion caused by PCV2 infection in mice. The findings may also prove useful in designing effective therapies against PCV2 infection.


Assuntos
Antioxidantes/farmacologia , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/metabolismo , Circovirus/patogenicidade , Glucanos/farmacologia , Fatores Imunológicos/farmacologia , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Infecções por Circoviridae/sangue , Infecções por Circoviridae/enzimologia , Citocinas/sangue , Feminino , Glutationa/biossíntese , Tolerância Imunológica/efeitos dos fármacos , Linfócitos/citologia , Linfócitos/efeitos dos fármacos , Masculino , Camundongos , Baço/citologia , Baço/efeitos dos fármacos , Baço/metabolismo , Superóxido Dismutase/metabolismo , Timo/citologia , Timo/efeitos dos fármacos , Timo/metabolismo
16.
Vaccine ; 29(22): 3923-9, 2011 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-21338683

RESUMO

The type III secretion system of Escherichia coli O157:H7 is involved in colonization of mammalian hosts by the organism. The translocated intimin receptor (Tir) is inserted into the mammalian host cell plasma membrane in a hairpin loop topology with the central loop of the molecule exposed to the host cell surface and accessible for interaction with an LEE-encoded bacterial outer membrane adhesin called intimin. Shiga toxin type 1 and 2 produced by E. coli O157:H7 are responsible for hemolytic uremic syndrome and able to promote intestinal colonization. Zonula occludens toxin (Zot) is a single polypeptide chain encoded by the filamentous bacteriophage CTXφ of Vibrio cholerae. Zot binds a receptor on intestinal epithelial cells and increases mucosal permeability by affecting the structure of epithelial tight junctions. Because of these properties, Zot is a promising tool for mucosal drug and antigen (Ag) delivery. In the current study, we constructed a novel fusion protein carrying both of the immunogenic B subunits derived from the two toxins, Tir and Zot, designated Stx2B-Tir-Stx1B-Zot, expressed in the E. coli BL21 and harvested the purified protein by a simple GST·Bind Resin chromatography method. We used a streptomycin-treated mouse model to evaluate the efficacy of subcutaneous vs. intranasal administration of the vaccine. Following immunization, mice were infected with E. coli O157:H7 and feces were monitored for shedding. Immune responses against Stx2B-Tir-Stx1B-Zot, Stx2B-Tir-Stx1B and control agent (GST/PBS) were also monitored. Subcutaneous immunization of mice with Stx2B-Tir-Stx1B-Zot induced significant Stx2B-Tir-Stx1B-Zot-specific serum IgG antibodies but did not significantly induce any antigen-specific IgA in feces, whereas intranasal immunization elicited significant Stx2B-Tir-Stx1B-Zot-specific serum IgG antibodies with some animals developing antigen-specific IgA in feces. Mice that were immunized intranasally with Stx2B-Tir-Stx1B-Zot showed dramatically decreased E. coli O157:H7 shedding compared to those of Stx2B-Tir-Stx1B and control agent following experimental infection. Mice immunized subcutaneously with Stx2B-Tir-Stx1B-Zot or Stx2B-Tir-Stx1B both showed reduced shedding in feces, moreover, Stx2B-Tir-Stx1B-Zot did better. These results demonstrate the perspective for the use of Stx2B-Tir-Stx1B-Zot to prevent colonization and shedding of E. coli O157:H7.


Assuntos
Toxina da Cólera/imunologia , Infecções por Escherichia coli/prevenção & controle , Escherichia coli O157/patogenicidade , Proteínas de Escherichia coli/imunologia , Vacinas contra Escherichia coli/imunologia , Receptores de Superfície Celular/imunologia , Toxina Shiga I/imunologia , Toxina Shiga II/imunologia , Administração Intranasal , Animais , Derrame de Bactérias/imunologia , Toxina da Cólera/genética , Endotoxinas , Infecções por Escherichia coli/imunologia , Escherichia coli O157/crescimento & desenvolvimento , Proteínas de Escherichia coli/genética , Vacinas contra Escherichia coli/administração & dosagem , Vacinas contra Escherichia coli/genética , Injeções Subcutâneas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Receptores de Superfície Celular/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Toxina Shiga I/genética , Toxina Shiga II/genética , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
17.
Acta Crystallogr Sect E Struct Rep Online ; 64(Pt 1): m152-3, 2007 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-21200505

RESUMO

In the centrosymmetric binuclear title complex, [Pb(2)(C(7)H(5)O(3))(4)(C(12)H(8)N(2))(2)(H(2)O)(2)]·H(2)O, each Pb atom is eight-coordinated in a PbO(6)N(2) environment by two N atoms from the 1,10-phenanthroline (phen) ligand, five carboxylate O atoms from four 3-hydroxy-benzoate anions and one O atom from the coordinated water mol-ecule in a distorted bicapped trigonal-prismatic geometry. The benzoate groups coordinate each Pb(II) atom in two different ways. Two benzoate ions behave as bidentate ligands to the Pb atom, and another benzoate ion bridges the Pb atoms, forming a binuclear structure. The dimeric units are packed via O-H⋯O hydrogen bonds and π-π inter-actions between the aromatic rings of neighboring mol-ecules, with centroid-centroid distances of 3.552 (2) and 3.641 (2) Å.

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