RESUMEN
Until recently, chemical pesticides were one of the most effective means of controlling agricultural pests; therefore, the search for insecticide targets for agricultural pests has been an ongoing problem. Estrogen-related receptors (ERRs) are transcription factors that regulate cellular metabolism and energy homeostasis in animals. Silkworms are highly sensitive to chemical pesticides, making them ideal models for pesticide screening and evaluation. In this study, we detected ERR expression in key organs involved in pesticide metabolism in silkworms (Bombyx mori), including the fat body and midgut. Using ChIP-seq technology, many estrogen- related response elements were identified in the 2000-bp promoter region upstream of metabolism-related genes, almost all of which were potential ERR target genes. The ERR inhibitor, XCT-790, and the endocrine disruptor, bisphenol A, significantly inhibited expression of the ERR target genes, BmTreh-1, BmTret-1, BmPK, BmPFK, and BmHK, in the fat bodies of silkworms, resulting in pupation difficulties in silkworm larvae that ultimately lead to death. In addition, based on the clarification that the ERR can bind to XCT-790, as observed through biofilm interferometry, its three-dimensional spatial structure was predicted, and using molecular docking techniques, small-molecule compounds with a stronger affinity for the ERR were identified. In summary, utilizing the powerful metabolic regulatory function of ERR in Lepidoptera pests, the developed small molecule inhibitors of ERR can be used for future control of Lepidoptera pests.
Asunto(s)
Bombyx , Simulación del Acoplamiento Molecular , Fenoles , Receptores de Estrógenos , Animales , Receptores de Estrógenos/metabolismo , Receptores de Estrógenos/genética , Bombyx/metabolismo , Bombyx/genética , Bombyx/efectos de los fármacos , Fenoles/farmacología , Compuestos de Bencidrilo/farmacología , Larva/metabolismo , Larva/efectos de los fármacos , Larva/genética , Insecticidas/farmacología , Proteínas de Insectos/metabolismo , Proteínas de Insectos/genética , Cuerpo Adiposo/metabolismo , Cuerpo Adiposo/efectos de los fármacos , Disruptores Endocrinos/farmacología , Disruptores Endocrinos/metabolismo , Nitrilos , TiazolesRESUMEN
Advances in the early detection of cancer are a way for treatment to be effective. However, due to the inability of early diagnose and treatment for lung cancer, the death rate of this type of cancer is high. Usually, a high percentage of patients are diagnosed at a stage where they are not able to receive treatment. The purpose of this study was the comparison of gene expression profiles in healthy individuals and people with lung cancer. The raw data sets GSE10072 and GSE19804 were taken from the GEO online database. Differentially expressed genes (DEGs) were identified between the non-tumor and tumor tissue samples using a meta-analysis investigation. Then, gene ontology and biological pathway analysis were performed with the Enrichr online server. The protein-protein interaction network of genes obtained from the meta-analysis investigation was drawn and analyzed using the String Online database and Cytoscape Software. Meta-analysis results showed a total of 515 differentially expressed genes. The results of the functional processes and biological pathway revealed that differentially expressed genes were mainly enriched in positive regulation of cell differentiation, regulation of cell population proliferation, regulation of epithelial cell differentiation, positive regulation of epithelial cell proliferation, response to growth factor, defense response to the tumor cell, cellular response to UV, regulation of cell cycle process, cell adhesion molecules, PPAR signaling pathway, TNF signaling pathway, ECM-receptor interaction, p53 signaling pathway, PI3K-Akt signaling pathway, and Cell cycle. Finally, key genes related to lung cancer, including IL6, MMP9, VWF, PECAM1, FOS, and CAV1 were identified. In conclusion, comparisons of gene expression profiles in healthy individuals and people with lung cancer identified some key genes that can act as lung cancer markers and can be used to predict new findings on cancer. These genes can play an important role in diagnosis and early cancer treatment.
RESUMEN
Development of supramolecular adhesives with strong tolerance to extreme conditions has emerged as an important research area. In this study, by balancing supramolecular interactions such as hydrogen bonding interactions, electrostatic interactions, π-π stacking interactions, and cation-π interactions, we designed and prepared a series of two-component supramolecular adhesives derived from small organic molecules. Highly efficient interfacial adhesion with maximum adhesion strength of ≈10.0â MPa was realized on various surfaces in air, organic solvents, or liquid nitrogen. Owing to balanced supramolecular interactions, water participation prolonged and increased the tolerance of the adhesives in extreme environments. We demonstrate that the combination of imidazole-based ionic liquids and phenols can be applied for various interfacial adhesions, thereby aiding the development of next-generation adhesives capable of adapting to various extreme conditions in a controlled manner.
RESUMEN
The present study aimed to investigate the effect of clopidogrel (CLO) on pharmacokinetics of ivabradine (IVA) and its metabolite in rats and develop a reliable method to determine IVA and its metabolite N-demethyl ivabradine in serum. Healthy male SD rats were randomized to be given 0.8 mg/kg IVA or IVA combined with 8 mg/kg CLO. Blood samples were collected at 0.083, 0.16, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 24 h after administration. The serum concentrations of IVA and N-demethyl ivabradine were determined by ultra-performance liquid chromatography-mass spectrometry and pharmacokinetic parameters were calculated using DASver3.0 software. The parameters of AUC(0 - t), AUC(0 - ∞), and Cmax for IVA in the group of IVA + CLO were significantly higher than those in the group of IVA (p < 0.01); the half-time (t1/2) in the IVA + CLO group was extended compared to IVA (p < 0.01) and CL/F was dropped obviously (p < 0.01). The decreases in AUC(0 - t), AUC(0 - ∞), and Cmax for N-demethyl ivabradine in the group of IVA + CLO was significantly compared to the group of IVA (p < 0.01). CL/F was higher than IVA (p < 0.01) and the t1/2 was slightly increased. In this study, we find that CLO restrains the metabolism of IVA into N-demethyl ivabradine, which may be related to its competitive inhibition effect on cytochrome P450 isoform 3A4(CYP3A4).
Asunto(s)
Benzazepinas/farmacocinética , Fármacos Cardiovasculares/farmacocinética , Inhibidores de Agregación Plaquetaria/farmacología , Ticlopidina/análogos & derivados , Animales , Área Bajo la Curva , Cromatografía Líquida de Alta Presión , Clopidogrel , Citocromo P-450 CYP3A/efectos de los fármacos , Citocromo P-450 CYP3A/metabolismo , Inhibidores del Citocromo P-450 CYP3A/farmacología , Interacciones Farmacológicas , Semivida , Ivabradina , Masculino , Espectrometría de Masas , Ratas , Ratas Sprague-Dawley , Ticlopidina/farmacologíaRESUMEN
Arboviruses can be paternally transmitted by male insects to offspring for long-term persistence, but the mechanism remains largely unknown. Here, we use a model system of a destructive rice reovirus and its leafhopper vector to find that insect ribosome-rescuer Pelo-Hbs1 complex expressed on the sperm surface mediates paternal arbovirus transmission. This occurs through targeting virus-containing tubules constituted by viral nonstructural protein Pns11 to sperm surface via Pns11-Pelo interaction. Tubule assembly is dependent on Hsp70 activity, while Pelo-Hbs1 complex inhibits tubule assembly via suppressing Hsp70 activity. However, virus-activated ubiquitin ligase E3 mediates Pelo ubiquitinated degradation, synergistically causing Hbs1 degradation. Importantly, Pns11 effectively competes with Pelo for binding to E3, thus antagonizing E3-mediated Pelo-Hbs1 degradation. These processes cause a slight reduction of Pelo-Hbs1 complex in infected testes, promoting effective tubule assembly. Our findings provide insight into how insect sperm-specific Pelo-Hbs1 complex is modulated to promote paternal virus transmission without disrupting sperm function.
Asunto(s)
Hemípteros , Proteínas de Insectos , Espermatozoides , Animales , Masculino , Espermatozoides/metabolismo , Espermatozoides/virología , Hemípteros/virología , Hemípteros/metabolismo , Proteínas de Insectos/metabolismo , Proteínas de Insectos/genética , Arbovirus , Proteínas HSP70 de Choque Térmico/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitina-Proteína Ligasas/genética , Reoviridae/fisiología , Insectos Vectores/virología , Insectos Vectores/metabolismo , Ribosomas/metabolismo , Infecciones por Arbovirus/transmisión , Infecciones por Arbovirus/metabolismo , Infecciones por Arbovirus/virologíaRESUMEN
Introduction: Signal peptide peptidase (SPP) is an intramembrane protease involved in a variety of biological processes, it participates in the processing of signal peptides after the release of the nascent protein to regulate the endoplasmic reticulum associated degradation (ERAD) pathway, binds misfolded membrane proteins, and aids in their clearance process. Additionally, it regulates normal immune surveillance and assists in the processing of viral proteins. Although SPP is essential for many viral infections, its role in silkworms remains unclear. Studying its role in the silkworm, Bombyx mori , may be helpful in breeding virus-resistant silkworms. Methods: First, we performed RT-qPCR to analyze the expression pattern of BmSPP. Subsequently, we inhibited BmSPP using the SPP inhibitor 1,3-di-(N-carboxybenzoyl-L-leucyl-L-leucylaminopropanone ((Z-LL)2-ketone) and downregulated the expression of BmSPP using CRISPR/Cas9 gene editing. Furthermore, we assessed the impact of these interventions on the proliferation of Bombyx mori nucleopolyhedrovirus (BmNPV). Results: We observed a decreased in the expression of BmSPP during viral proliferation. It was found that higher concentration of the inhibitor resulted in greater inhibition of BmNPV proliferation. The down-regulation of BmSPP in both in vivo and in vitro was found to affect the proliferation of BmNPV. In comparison to wild type silkworm, BmSPPKO silkworms exhibited a 12.4% reduction in mortality rate. Discussion: Collectively, this work demonstrates that BmSPP plays a negative regulatory role in silkworm resistance to BmNPV infection and is involved in virus proliferation and replication processes. This finding suggests that BmSPP servers as a target gene for BmNPV virus resistance in silkworms and can be utilized in resistance breeding programs.
Asunto(s)
Bombyx , Nucleopoliedrovirus , Animales , Nucleopoliedrovirus/genética , Edición Génica , Regulación hacia AbajoRESUMEN
Introduction: Remimazolam is an ultra-short-acting benzodiazepine sedative agent commonly used in general anesthesia, procedural sedation, and intensive care unit (ICU) sedation. This study aimed to explore the efficacy and safety of remimazolam versus propofol for the induction and maintenance of general anesthesia in preschool-age children undergoing elective surgery. Methods and analysis: In this multicenter, randomized, single-blind, positive-controlled non-inferior clinical trial, one hundred ninety-two children aged 3-6 years will be randomly allocated as a 3:1 ratio into two groups: Group R with an intravenous dose of remimazolam 0.3 mg/kg for the induction of anesthesia followed by a constant infusion rate of remimazolam 1-3 mg/kg/h to maintain anesthesia, and Group P with an intravenous dose of propofol 2.5 mg/kg for the induction of anesthesia followed by a constant infusion rate of propofol 4-12 mg/kg/h to maintain anesthesia. The primary outcome will be the rate of the successful induction and maintenance of anesthesia. The secondary outcomes will include the time to LoC, the Bispectral Index (BIS) value, awakening time, extubation time, post-anesthesia care unit (PACU) discharge time, usage of additional sedative drugs during the induction period, usage of remedial drugs in PACU, emergence delirium, pain in PACU, behavior scores at day 3 after surgery, parental and anesthesiologists' satisfaction, and adverse events. Ethics and dissemination: This study has been approved by the ethics review boards at all participating hospitals. The Ethics Committee of the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University (Reference No. LCKY 2020-380, November 13, 2020) is the central ethics committee.
RESUMEN
OBJECTIVE: CD-1 was isolated from aquatic environment, which grew with strict L-cysteine dependence. In this study, we applied molecular methods to identify CD-1, and animal test to understand its virulence. METHODS: To identify CD-1 strain, CD-1 strain was tested for genus-specific 16S rRNA of Legionella via PCR amplification, then its rpoB gene was sequenced for phylogenic analysis. To understand the virulence, CD-1 was detected for mip gene, which was an indispensable virulent gene of Legionella. Then, BABL/c mice were infected by CD-1 in different dosages. RESULTS: For identification, CD-1 was positive for genus-specific 16S rRNA of Legionella, while in the phylogenic tree CD-1 was a sister to Legionella longbeachae with high posterior probability (PP = 1.00). For the virulence analysis, CD-1 was positive for mip gene detection. In the animal test, all mice tested died when the infection dose of CD-1 strain reached 10(7) cfu/mL. CONCLUSION: CD-1 strain was identified to be Legionella longbeachae with strong virulence to BALB/c mice. It may be a potential virulent strain to human. This is the first strain of Legionella longbeachae isolated in Sichuan province, and this is the first virulence analysis of Legionella strain isolated from aquatic environment in China.
Asunto(s)
Legionella longbeachae/aislamiento & purificación , Legionella longbeachae/patogenicidad , Virulencia , Microbiología del Agua , Animales , China , Legionella longbeachae/genética , Masculino , Ratones , Ratones Endogámicos BALB C , Filogenia , ARN Ribosómico 16S/genéticaRESUMEN
Objectives: Although lipids have been assessed for their possible roles in cancer survival prediction, studies on the association between serum triglyceride (TG) levels and the prognosis of esophageal squamous cell carcinoma (ESCC) patients are limited. This study aimed to evaluate whether serum TG is associated with outcomes in patients with ESCC and investigate any interaction between serum TG and clinical parameters, especially body fat mass. Materials and methods: We conducted a prospective case study on patients diagnosed with ESCC between March 2012 and November 2018. We measured patients' serum TG levels before and after treatment. The association between serum TG and overall survival (OS) was evaluated using hazard ratios. We sought to determine a threshold point using optimal stratification. Survival analysis was performed using Kaplan-Meier curves and a Cox proportional hazards model. Results: Of the 257 participants diagnosed with ESCC, 200 (77.8%) were men. Median follow-up time was 22.4 months (range 3.3-92.4 months). Using univariate Cox proportional hazard analysis and subsequent multivariate analysis, post-TG levels, Karnofsky performance scores, T stages, and chemotherapy cycles were shown to be independent prognostic factors for OS (p < 0.05). The post-TG cut-off point to best classify patients with respect to time to mortality was 1.47 mmol/L. A post-TG level of ≥ 1.47 mmol/L could independently predict a better OS (hazard ratio: 0.55, 95% confidence interval: 0.37-0.79). The associations were consistent across the subtypes of clinical parameters. Furthermore, the post-body mass index, post-subcutaneous adipose tissue area, post-visceral adipose tissue area, post-total adiposity tissue area, and post-total adipose density exhibited a strong positive association with post-TG levels. Conclusion: Post-TG levels were found to be a significant positive prognostic biomarker for body fat mass and OS in ESCC patients.
RESUMEN
Interfacial adhesion under extreme conditions has attracted increasing attention owing to its potential application of stopping leakages of oil or natural gas. However, interfacial adhesion is rarely stable at ultralow temperatures and in organic solvents, necessitating the elucidation of the molecular-level processes. Herein, we used the intermolecular force-control strategy to prepare four linear polymers by tuning the proportion of hydrogen bonding and the number of electrostatic sites. The obtained polymeric ion liquids displayed strong dynamic adhesion at various interfaces. They also efficiently tolerated organic solvents and ultracold temperatures. Highly reversible rheological behaviors are observed within a thermal cycle between high and ultracold temperatures. Temperature-dependent infrared spectra and theoretical calculation reveal thermal reversibility and interfacial adhesion/debonding processes at the molecular level, respectively. This intermolecular force-control strategy may be applied to produce environmentally adaptive functional materials for real applications.
RESUMEN
Cystic echinococcosis (CE), a devastating zoonotic condition caused by the tapeworm Echinococcus granulosus, remain a significant public health problem worldwide. However, after a negative correlation between solid tumor and CE has been incidentally discovered, accumulating evidence have suggested that this parasite may induce anticancer effect through activating host immune response and secreting molecules with anticancer potential, which may provide some new understanding for immunotherapy. This article will review the evidence supporting the anticancer effect of E. granulosus and its underlying mechanisms and discuss the possible implications in immunotherapy.
RESUMEN
BACKGROUND: General anesthesia combining with a caudal block (CB) has been commonly performed in pediatric patients undergoing circumcision surgeries. However, some severe complications have been suspected of a caudal block in the combined use. To avoid these issues of a caudal block, this study introduces a novel dorsal penile nerve block (DPNB) via perineum guided by ultrasound as an alternative to a caudal block in pediatric circumcision surgeries. METHODS: A total of 104 pediatric patients scheduled for circumcision surgeries were involved and randomly divided into 2 groups: the CB group (n=52) and the DPNB group (n=52). A laryngeal mask was inserted followed by induction and maintenance anesthesia of inhaled sevoflurane. In the DPNB group, a dorsal penile nerve block (DPNB) guided by a real-time ultrasonography was performed by a single injection via perineum of 0.25% ropivacaine plus 0.8% lidocaine with total injection volume of 3-5ml. In the CB group, a dose of 0.5 ml/kg was given via the caudal canal following the same general anesthesia with that of Group DPNB. The time to the first analgesic demand after surgery is the key data collected for comparing between the two study groups. Heart rates and respiratory rates changes before and during the surgical procedure, pain score when leaving the PACU, and the time taken for the first micturition after a surgery were also recorded to analyze the differences in analgesic effects between the CB and DPNB groups. RESULTS: No significant difference in heart rates and respiratory rates was found between the two groups before and during the surgery. Pain scores were similar before pediatric patients leave the PACU. However, the time taken for the first micturition after a surgery in Group DPNB is shorter than Group CB. The patients in Group DPNB asked for analgesics later than those in Group CB. Additionally, no significant differences in adverse effects were noted between two groups except the numbness of the lower limbs occurring less in Group DPNB. CONCLUSIONS: The ultrasound-guided dorsal penile nerve block via perineal approach can basically act as a safe and effective alternative to the caudal block in pediatric patients undergoing circumcision surgeries. Clinical Trials identifier is ChiCTR-IPR-15006670. Protocol is available at http://www.chictr.org.cn/showproj.aspx?proj=11319.
Asunto(s)
Anestesia General/métodos , Circuncisión Masculina/métodos , Pene/cirugía , Nervio Pudendo/efectos de los fármacos , Niño , Preescolar , Humanos , Lidocaína/administración & dosificación , Masculino , Bloqueo Nervioso/métodos , Dimensión del Dolor/métodos , Pene/inervación , Ropivacaína/administración & dosificaciónRESUMEN
Legionellosis (Legionnaires' disease; LD) is a form of severe pneumonia caused by species of Legionella bacteria. Because inhalation of Legionella-contaminated aerosol is considered the major infection route, routine assessments of potential infection sources such as hot water systems, air-conditioner cooling water, and municipal fountains are of great importance. In this study, we utilized in vitro culture and multilocus sequence analysis (MLSA) targeting 16S rRNA, mip, rpoB, and mip-rpoB concatenation to isolate and identify Legionella spp. from 5 municipal fountains in Chengdu City, Sichuan Province, China. Our results demonstrated that 16S rRNA was useful for initial identification, as it could recognize isolates robustly at the genus level, while the genes mip, rpoB, and mip-rpoB concatenation could confidently discriminate Legionella species. Notably, the three subspecies of L. pneumophila could be distinguished by the analysis based on rpoB. The serotyping result of strain CD-1 was consistent with genetic analysis based on the concatenation of mip and rpoB. Despite regular maintenance and sanitizing methods, 4 of the 5 municipal fountains investigated in this study were positive for Legionella contamination. Thus, regularly scheduled monitoring of municipal fountains is urgently needed as well as vigilant disinfection. Although the application of MLSA for inspection of potential sites of infection in public areas is not standard procedure, further investigations may prove its usefulness.
Asunto(s)
Proteínas Bacterianas/genética , ARN Polimerasas Dirigidas por ADN/genética , Legionella/clasificación , Legionella/aislamiento & purificación , Tipificación de Secuencias Multilocus/métodos , Isomerasa de Peptidilprolil/genética , ARN Ribosómico 16S/genética , Microbiología del Agua , China , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Genotipo , Humanos , Legionella/genética , Datos de Secuencia MolecularRESUMEN
The leishmaniases are zoonotic diseases caused by protozoan parasites of the genus Leishmania. Leishmaniases are still endemic in China, especially in the west and northwest froniter regions. To revalue the preliminary phylogenetic results of Chinese Leishmania isolates, we amplified partial fragment of small subunit ribosomal RNA (SSU rRNA) and 7 spliced leader RNA (7SL RNA), then tested the phylogenetic relationships among Chinese Leishmania isolates and their relatives by analyzing SSU rRNA gene sequences and 7SL RNA gene sequences. 19 SSU RNA sequences and 9 7SL RNA sequences were obtained in our study, then analyzed with 42 SSU RNA sequences and 32 7SL RNA sequences retrieved from Genbank, respectively. In the Bayesian analysis of the SSU RNA gene, the isolate MHOM/CN/93/GS7 and the isolate IPHL/CN/77/XJ771 are members of Leishmania donovani complex, while the isolate MHOM/CN/84/JS1 clustered with Leishmania tropica. The other 11 Chinese Leishmania isolates (MHOM/CN/90/WC, MCAN/CN/90/SC11, MHOM/CN/80/XJ801, MHOM/CN/85/GS4, MHOM/CN/84/SD1, MCAN/CN/86/SC7, MHOM/CN/54/#3, MHOM/CN/83/GS2, MHOM/CN/90/SC10H2, MHOM/CN/89/GS6 and MHOM/CN/ 89/GS5) form an unclassified group, defined as Leishmania sp., and the most relative species to this group is L. tarentolae. In the Bayesian analysis of the 7SL RNA gene, 9 Chinese Leishmania isolates also formed an unclassified group with L. tarentolae, including canine isolate 10, MHOM/CN/85/GS4, MHOM/CN/84/SD1, MCAN/CN/86/SC7, MHOM/CN/54/#3, MHOM/ CN/83/GS2, MHOM/CN/90/SC10H2, MHOM/CN/89/GS6 and MHOM/CN/89/GS5. We concluded that: (1) Chinese Leishmania isolates are non-monophyly group; (2) an unclassified group may exist in China, and the most relative species to this group is L. tarentolae; (3) MHOM/CN/84/JS1, which was previously assigned as L. donovani, was most genetically related to L. tropica strain MHOM/SU/74/K27.
Asunto(s)
Leishmania/genética , ARN Protozoario/genética , ARN Ribosómico/genética , Animales , Secuencia de Bases , China , Citocromos/genética , Regulación de la Expresión Génica/fisiología , FilogeniaRESUMEN
BACKGROUND: Leishmaniasis is a vector-borne disease, which is still endemic in the west and northwest area of China. Canines are the major reservoirs of Leishmania, the etiological agent of human visceral leishmaniasis. Phlebotomus chinensis is the main transmission vector of zoonotic visceral leishmaniasis (ZVL). METHODS: In this study, rK39 dip-stick, ELISA and PCR methods were used to investigate the prevalence of canine leishmaniasis (CanL) in Beichuan County, Sichuan Province, China. RESULTS: Among the 86 dogs which were included in the study, 13 dogs were positive using the dip-stick test (15.12%), while 8 dogs were positive using ELISA (9.30%) and 19 dogs were positive for PCR (22.03%). In total, 32 dogs were positive for one or more tests (37.21%). Interestingly, phylogenetic analysis based on the partial 7SL RNA fragment provided evidence that an undescribed Leishmania species, which is clearly a causative agent of CanL and human visceral leishmaniasis, does exist in China. This result is consistent with our previous study. CONCLUSIONS: Our work confirmed that canine leishmaniasis is still prevalent in Beichuan County. Further control is urgently needed, as canine leishmaniasis is of great public health importance. The phylogenetic analysis based on 7SL RNA segment provides evidence for the existence of an undescribed Leishmania sp. in China.
Asunto(s)
Enfermedades de los Perros/parasitología , Leishmania/aislamiento & purificación , Leishmaniasis Visceral/veterinaria , ARN Citoplasmático Pequeño/análisis , Partícula de Reconocimiento de Señal/análisis , Animales , China/epidemiología , Enfermedades de los Perros/epidemiología , Perros , Enfermedades Endémicas , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Insectos Vectores/parasitología , Leishmania/genética , Leishmaniasis Visceral/epidemiología , Leishmaniasis Visceral/parasitología , Masculino , Filogenia , Reacción en Cadena de la Polimerasa , Prevalencia , Tiras Reactivas , Zoonosis/epidemiología , Zoonosis/parasitologíaRESUMEN
To investigate the protect effects of the recombinant protein FlaA/MompS/PilE against Legionella pneumophila (L. pneumophila), the coding sequences of the three proteins were optimized by DNA Star software firstly, cloned, expressed by Escherichia coli BL21, and purified. To give an enhanced the immunological response, the proteins were linked together with (Linker) or without a linker insert (NLinker) and were purified from E. coli BL21. The A/J mouse model was used to determine the level of the induction of protective immunity from the purified proteins. Our results showed that the IgG titer, which was measured by ELISA, was increased after the administration of the five proteins. Compared to the administration of the individual proteins, the chimeric Linker and NLinker proteins displayed lasting immunity to a lethal dose of L. pneumophila challenge. The Linker protein protected the A/J mouse against a higher dose of L. pneumonia compared to the other proteins used in this study, as it contained a more effective immunogen. The work presented here demonstrates that the bioinformatics software, DNA Star, is a valid tool to analyse the epitopes of proteins and was useful in the optimization of proteins that could induce the protective immune response to L. pneumophila. The cross-immunity of recombinant proteins, such as the Linker and the NLinker chimera, have higher generates a greater immune than the single proteins.
Asunto(s)
Proteínas Bacterianas/inmunología , Proteínas Fimbrias/inmunología , Flagelina/inmunología , Inmunoglobulina G/sangre , Legionella pneumophila/inmunología , Enfermedad de los Legionarios/prevención & control , Porinas/inmunología , Proteínas Recombinantes/inmunología , Animales , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Secuencia de Bases , Biología Computacional/métodos , Reactivos de Enlaces Cruzados , Mapeo Epitopo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Fimbrias/química , Proteínas Fimbrias/genética , Proteínas Fimbrias/metabolismo , Flagelina/química , Flagelina/genética , Flagelina/metabolismo , Inmunidad , Inmunización , Legionella pneumophila/patogenicidad , Enfermedad de los Legionarios/inmunología , Enfermedad de los Legionarios/mortalidad , Ratones , Datos de Secuencia Molecular , Porinas/química , Porinas/genética , Porinas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Programas InformáticosRESUMEN
OBJECTIVE: To construct the eukaryotic expression plasmid containing lvgA gene flanked with CpG motifs of Legionella pneumophila for its expression in NIH3T3 cells. METHODS: lvgA gene flanked with CpG motifs of Legionella pneumophila was amplified by PCR. The PCR products was inserted into the eukaryotic expression plasmid pcDNA3.1/myc-his(+) to construct the recombinant plasmid pclvgA/CpG, which was subsequently transfected into NIH3T3 cells via lipofection. Immunofluorescence analysis was carried out to detect the transient expression of the plasmid in the cells. RESULTS: Sequence analysis showed that the recombinant plasmid pclvgA/CpG contained the lvgA/CpG fragment with a length of 657 bp, encoding a protein of 27.7 Ku. Immunofluorescence analysis identified the transient expression of the recombinant plasmid pclvgA/CpG in NIH3T3 cells. CONCLUSION: The lvgA gene flanked with CpG motifs of Legionella pneumophila has been constructed successfully, and the transient expression of the recombinant plasmid pclvgA/CpG can be detected in NIH3T3 cells.