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1.
Discov Med ; 36(187): 1743-1757, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-39190389

RESUMEN

BACKGROUND: Cerebral ischemia-reperfusion injury (CIRI) is a prevalent neurological disorder, characterized by the oxidative stress and inflammatory response induced during the ischemia-reperfusion process, leading to significant damage to brain cells. Ginsenoside Rb1, a natural medicinal ingredient, possesses potential neuroprotective effects. This study aims to investigate the mechanism of action of ginsenoside Rb1 in CIRI and its protective effects on brain injury. METHODS: We utilized a mouse CIRI model and randomly divided the mice into control group, CIRI group, and ginsenoside Rb1 treatment group. The effects of Rb1 on brain tissue damage, apoptosis, expression of inflammatory factors, and pyroptotic cell numbers in CIRI mice were observed through triphenyl tetrazolium chloride (TTC) staining, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, real-time reverse transcription polymerase chain reaction (qRT-PCR), and electron microscopy. In a cell model, the regulatory effect of Rb1 on oxygen-glucose deprivation/reperfusion (OGD/R)-induced HT22 cell pyroptosis via the nuclear respiratoty factor 2/tumor necrosis factor-α (TNF-α)-induced Protein 3 (TNFAIP3, aka A20)/eukaryotic translation elongation factor 1A2 (Nrf2/A20/eEF1A2) axis was detected using Western blot and TUNEL staining. Additionally, the impact of Nrf2 inhibitor ML385 and eEF1A2 overexpression on the neuroprotective effect of Rb1 was assessed. Using the comprehensive experimental methods mentioned above, the neuroprotective mechanism of Rb1 in CIRI was thoroughly evaluated. RESULTS: Our findings demonstrate that treatment with ginsenoside Rb1 alleviated behavioral deficits induced by CIRI and reduced pathological damage in brain tissue. Furthermore, ginsenoside Rb1 treatment notably decreased oxidative stress and the inflammatory response induced by CIRI, leading to lower levels of inflammatory factors (p < 0.05). Further experimental results indicated that ginsenoside Rb1 promoted antioxidant and anti-inflammatory responses by regulating the activity of the Nrf2/A20/eEF1A2 axis. Additionally, ginsenoside Rb1 inhibited the activation of the NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome, thereby reducing the release of inflammatory factors and the occurrence of cell apoptosis. CONCLUSION: Our study results suggest that ginsenoside Rb1 exerts neuroprotective effects and alleviates brain injury induced by CIRI by regulating the Nrf2/A20/eEF1A2 axis and inhibiting the activation of the NLRP3 inflammasome. These findings provide new treatment insights for CIRI and support ginsenoside Rb1's development as a therapeutic drug. However, despite the promising nature of our findings, further research is required to validate these discoveries and explore the feasibility and safety of ginsenoside Rb1 in clinical applications. We hope that our study can provide new directions and strategies for the treatment and prevention of CIRI, contributing to the development of neuroprotective drugs.


Asunto(s)
Ginsenósidos , Factor 2 Relacionado con NF-E2 , Daño por Reperfusión , Ginsenósidos/farmacología , Ginsenósidos/uso terapéutico , Animales , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Daño por Reperfusión/tratamiento farmacológico , Factor 2 Relacionado con NF-E2/metabolismo , Ratones , Masculino , Transducción de Señal/efectos de los fármacos , Modelos Animales de Enfermedad , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Ratones Endogámicos C57BL , Isquemia Encefálica/metabolismo , Isquemia Encefálica/tratamiento farmacológico , Isquemia Encefálica/patología , Estrés Oxidativo/efectos de los fármacos , Apoptosis/efectos de los fármacos , Línea Celular
2.
Int J Chron Obstruct Pulmon Dis ; 19: 1491-1513, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38957709

RESUMEN

Background: Chronic obstructive pulmonary disease (COPD) stands as a predominant cause of global morbidity and mortality. This study aims to elucidate the relationship between pyroptosis-related genes (PRGs) and COPD diagnosis in the context of immune infiltration, ultimately proposing a PRG-based diagnostic model for predicting COPD outcomes. Methods: Clinical data and PRGs of COPD patients were sourced from the GEO database. The "ConsensusClusterPlus" package was employed to generate molecular subtypes derived from PRGs that were identified through differential expression analysis and LASSO Cox analysis. A diagnostic signature including eight genes (CASP4, CASP5, ELANE, GPX4, NLRP1, GSDME, NOD1and IL18) was also constructed. Immune cell infiltration calculated by the ESTIMATE score, Stroma scores and Immune scores were also compared on the basis of pyroptosis-related molecular subtypes and the risk signature. We finally used qRT - PCR to detect the expression levels of eight genes in COPD patient and normal. Results: The diagnostic model, anchored on eight PRGs, underwent validation with an independent experimental cohort. The area under the receiver operating characteristic (ROC) curves (AUC) for the diagnostic model showcased values of 0.809, 0.765, and 0.956 for the GSE76925, GSE8545, and GSE5058 datasets, respectively. Distinct expression patterns and clinical attributes of PRGs were observed between the comparative groups, with functional analysis underscoring a disparity in immune-related functions between them. Conclusion: In this study, we developed a potential as diagnostic biomarkers for COPD and have a significant role in modulating the immune response. Such insights pave the way for novel diagnostic and therapeutic strategies for COPD.


Asunto(s)
Bases de Datos Genéticas , Valor Predictivo de las Pruebas , Enfermedad Pulmonar Obstructiva Crónica , Piroptosis , Humanos , Enfermedad Pulmonar Obstructiva Crónica/genética , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico , Enfermedad Pulmonar Obstructiva Crónica/inmunología , Piroptosis/genética , Perfilación de la Expresión Génica , Pulmón/inmunología , Masculino , Femenino , Persona de Mediana Edad , Marcadores Genéticos , Estudios de Casos y Controles , Transcriptoma , Anciano , Reproducibilidad de los Resultados , Predisposición Genética a la Enfermedad , Pronóstico
3.
Infect Immun ; 91(12): e0027323, 2023 Dec 12.
Artículo en Inglés | MEDLINE | ID: mdl-37929972

RESUMEN

Trueperella pyogenes can cause severe pulmonary disease in swine, but the mechanism of pathogenesis is not well defined. T. pyogenes-induced damage to porcine bronchial epithelial cells (PBECs), porcine precision-cut lung slices (PCLS), and respiratory epithelium of mice remains unknown. In this study, we used T. pyogenes 20121 to infect PBECs in air-liquid interface conditions and porcine PCLS. T. pyogenes could adhere to, colonize, and induce cytotoxic effect on PBECs and the luminal surface of bronchi in PCLS, which damaged the bronchiolar epithelium. Moreover, bronchiolar epithelial cells showed extensive degeneration in the lungs of infected mice. Furthermore, western blot showed that the NOD-like receptor (NLR)/C-terminal caspase recruitment domain (ASC)/caspase-1 axis and nuclear factor-kappa B pathway were involved in inflammation in PCLS and lungs of mice, which also confirms that porcine PCLS provide a platform to analyze the pulmonary immune response. Meanwhile, the levels of p-c-Jun N-terminal kinase, p-extracellular signal-regulated kinase, and p-protein kinase B (AKT) were increased significantly, which indicated the mitogen-activated protein kinase and Akt pathways were also involved in inflammation in T. pyogenes-infected mice. In addition, we used T. pyogenes 20121 to infect tumor necrosis factor-alpha (tnf-α-/-) mice, and the results indicated that apoptosis and injury in respiratory epithelium of infected tnf-α-/- mice were alleviated. Thus, the pro-inflammatory cytokine TNF-α played a role in apoptosis and the respiratory epithelium injury in mouse lungs. Collectively, our study provides insight into the inflammatory injury induced by T. pyogenes and suggests that blocking NLR may be a potential therapeutic strategy against T. pyogenes infection.


Asunto(s)
Proteínas Proto-Oncogénicas c-akt , Factor de Necrosis Tumoral alfa , Animales , Ratones , Porcinos , Inflamación , Epitelio/patología , Citocinas
4.
Vet Sci ; 10(8)2023 Aug 05.
Artículo en Inglés | MEDLINE | ID: mdl-37624293

RESUMEN

Serological testing is an important method for the diagnosis of pseudorabies virus (PRV) infection. We aimed to investigate the envelope glycoprotein I (gI) of PRV, a strong immunogen, and its potential as an efficient and low-cost diagnostic reagent. In this study, the DNA of the PRV SC strain was used as the template, and the recombinant fragment of gI (633 bp) was amplified via PCR using synthetic primers, and was then ligated into the pET-30a expression vector. The constructs were transferred into Escherichia coli (E. coli) for prokaryotic expression, and the antigenicity of the expression products was identified by Western blot analysis with pig positive serum against PRV. The recombinant protein was purified by a Ni column, and BALB/c mice were immunized with purified gI protein to obtain anti-gI-positive serum. After PK-15 cells had been infected by PRV for 48 h, the immunogenicity of purified gI protein was identified with a fluorescence immunoassay using anti-gI mouse serum. The recombinant plasmid (pET-30a-gI) was expressed, and the native gI protein was obtained after denaturation by urea and renaturation by dialysis. A small-scale ELISA test containing 1.0 µg/mL of purified gI protein was designed to evaluate pig serum (80 samples), and the results of the ELISA test were compared to those of competitive ELISA (cELISA) tests using IDEXX Kits, which resulted in 97.5% consistency. The results suggested that the truncated gI protein may be a potential diagnostic reagent.

5.
Animals (Basel) ; 13(7)2023 Apr 03.
Artículo en Inglés | MEDLINE | ID: mdl-37048495

RESUMEN

In China, the national-level protected pig, the Min pig, is characterized by the development of secondary hairs and hair follicles in winter. Factors that dominate the genotype in the growth of secondary hairs are not clear through the concrete cell signaling pathways. This study compared hair phenotypes based on morphological structure, transcriptomics, and potential targeting molecules in the breeds of Min, Berkshire, and Yorkshire pigs. The results indicated that Min pigs have specific characteristics for the growth of secondary hairs compared with the Berkshire and Yorkshire pigs. The transcriptome analyses and quantitative reverse transcription-polymerase chain reaction results revealed that secondary hair growth was activated by follicle stem cells. The specific inhibitors of Wnt and BMP were studied using respective signals. The density of follicles, activity of follicle stem cells, and relative gene expression results have shown that Wnt and BMP stimulate the activity of follicle stem cells, and the Wnt signaling molecule has a significantly better effect than the BMP signaling molecule on stem cells. Wnt and BMP can promote the growth of local secondary hair and gene expression. Therefore, this study was conducted to verify the development mechanisms of secondary hairs, which have potential applications in laboratory animals and comparative medicine.

6.
Front Neurol ; 12: 706631, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34858306

RESUMEN

Purpose: To investigate the brain information flow pattern in patients with early mild cognitive impairment (EMCI) and explore its potential ability of differentiation and prediction for EMCI. Methods: In this study, 49 patients with EMCI and 40 age- and sex-matched healthy controls (HCs) with available resting-state functional MRI images and neurological measures [including the neuropsychological evaluation and cerebrospinal fluid (CSF) biomarkers] were included from the Alzheimer's Disease Neuroimaging Initiative. Functional MRI measures including preferred information flow direction between brain regions and preferred information flow index of each brain region parcellated by the Atlas of Intrinsic Connectivity of Homotopic Areas (AICHA) were calculated by using non-parametric multiplicative regression-Granger causality analysis (NPMR-GCA). Edge- and node-wise Student's t-test was conducted for between-group comparison. Support vector classification was performed to differentiate EMCI from HC. The least absolute shrinkage and selection operator (lasso) regression were used to evaluate the predictive ability of information flow measures for the neurological state. Results: Compared to HC, disturbed preferred information flow directions between brain regions involving default mode network (DMN), executive control network (ECN), somatomotor network (SMN), and visual network (VN) were observed in patients with EMCI. An altered preferred information flow index in several brain regions (including the thalamus, posterior cingulate, and precentral gyrus) was also observed. Classification accuracy of 80% for differentiating patients with EMCI from HC was achieved by using the preferred information flow directions. The preferred information flow directions have a good ability to predict memory and executive function, level of amyloid ß, tau protein, and phosphorylated tau protein with the high Pearson's correlation coefficients (r > 0.7) between predictive and actual neurological measures. Conclusion: Patients with EMCI were presented with a disturbed brain information flow pattern, which could help clinicians to identify patients with EMCI and assess their neurological state.

7.
Pathogens ; 10(2)2021 Feb 16.
Artículo en Inglés | MEDLINE | ID: mdl-33669225

RESUMEN

Streptococcus suis causes disease in pigs and is implicated increasingly in human disease worldwide. Although most clinical cases are associated with serotype 2, infections by other serotypes have sometimes been reported. Here, we sequenced the genome of a multidrug-resistant S. suis serotype 28 (strain 11313) and a multidrug-resistant S. suis serotype 31 (strain 11LB5). Strain 11313 was apathogenic in mouse infection models, whereas strain 11LB5 displayed ganglion demyelination, meningeal thickening, congestion, mononuclear cell infiltration, massive proliferation of cortical glial cells, and bacteria (>104 CFU/g) in the spinal cord and ganglia in mice. Furthermore, immunohistochemistry found that the heavily infiltrated glial cells were astrocytes. Strain 11313 harbored the resistance genes ant(6)-Ia, erm(B), optrA, tet(l), tet(o), and strain 11LB5 harbored the resistance genes ant(6)-Ia, erm(B), tet(40), tet(o/w/32/o), aac(6')-aph(2″). Mouse studies showed that strain 11LB5 exhibited a similar virulence to serotype 2 strain 700794, highlighting the need for surveillance of the other serotype S. suis isolates, in addition to serotype 2, in farms. This is the first report of the aminoglycoside resistance gene ant(6)-Ia in S. suis from animals. This suggests that S. suis might serve as an antibiotic resistance reservoir, which spreads the resistance gene ant(6)-Ia or optrA to other streptococcal pathogens on farms.

8.
Vaccines (Basel) ; 9(2)2021 Feb 08.
Artículo en Inglés | MEDLINE | ID: mdl-33567652

RESUMEN

Canine adenovirus (CAdV) has a high prevalence in canine populations. High affinity neutralizing antibodies against conserved epitopes can provide protective immunity against CAdV and protect against future outbreaks. In this study, we identified two CAdV-2-specific neutralizing monoclonal antibodies (mAbs), 2C1 and 7D7, which recognized two linear-dependent epitopes. MAb 2C1 potently neutralized CAdV-2 with a 50% neutralization titer (NT50) of 4096, and mAb 7D7 partially neutralized CAdV-2 with a 50% NT50 of 64. Immunoprecipitation, Western blot and protein spectral analysis indicated that both neutralizing mAbs recognized the hexon protein (Hex) of CAdV-2. Through a 12-mer random peptide phage display and synthetic peptides analysis, we finely mapped the neutralizing epitopes to two 10-amino acid (aa) peptides within the CAdV Hex: 634RIKQRETPAL643 located on the surface region; and 736PESYKDRMYS745 located in the inner region of the expected 3D structure of trimeric Hex. Importantly, the two epitopes are highly conserved among all CAdV isolates by sequence alignment analysis. Thus, these results provide insights into the interaction between virus and mAbs at the aa level and may have potential applications in the development of novel therapeutic or epitope-based vaccines, antibody therapeutics and a diagnostic method suitable for the rapid detection of all CAdVs.

9.
Front Vet Sci ; 8: 824349, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-35174243

RESUMEN

Trueperella pyogenes causes disease in cattle, sheep, goats and swine, and is involved occasionally in human disease worldwide. Most reports implicating T. pyogenes have been associated with clinical cases, whereas no report has focused on pathogenicity of T. pyogenes in mouse models or precision-cut lung slice (PCLS) cultures from swine. Here, we isolated and identified a virulent, ß-hemolytic, multidrug-resistant T. pyogenes strain named 20121, which harbors the virulence marker genes fimA, fimE, nanH, nanP and plo. It was found to be highly resistant to erythromycin, azithromycin and medemycin. Strain 20121 was pathogenic in mouse infection models, displaying pulmonary congestion and inflammatory cell infiltration, partial degeneration in epithelial cells of the tracheal and bronchiolar mucosa, a small amount of inflammatory cell infiltration in the submucosa, and bacteria (>104 CFU/g) in the lung. Importantly, we used T. pyogenes 20121 to infect porcine precision-cut lung slices (PCLS) cultures for the first time, where it caused severe bronchoconstriction. Furthermore, dexamethasone showed its ability to relieve bronchoconstriction in PCLS caused by T. pyogenes 20121, highlighting dexamethasone may assist antibiotic treatment for clinical T. pyogenes infection. This is the first report of T. pyogenes used to infect and cause bronchoconstriction in porcine PCLS. Our results suggest that porcine PCLS cultures as a valuable 3D organ model for the study of T. pyogenes infection and treatment in vitro.

10.
Poult Sci ; 97(11): 3899-3909, 2018 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-29982682

RESUMEN

To explore the relationship among the level of fiber, gut microbiota, and nutritional substances, we applied the next generation sequencing technology for the identification of the composition and structure of microbiota in the gastrointestinal tract. In this study, a total of 25 phyla and 298 genera were identified from the gastrointestinal tract; Firmicutes, Bacteroidetes, and Proteobacteria were the predominant phyla. The ability of cecum in carbohydrate metabolism was significantly higher than that of the gizzard and ileum (P < 0.05). The bacterial community structure in various stages of the development of the cecum was different. In the different growth stages of cecum, the increase in the microbiota structure of the fiber level elevates the ability of carbon hydration. Second, the apparent metabolic rates of the other nutrients were affected by the fiber and period except for acid detergent fiber (P < 0.05); the apparent utilization rate of the nutrients increased with time. However, with the increase in the fiber level, the apparent utilization of nutrients was initially increased, followed by a decrease. Therefore, a correlation was established between the fiber level and gastrointestinal microbiota and apparent nutrient utilization rate of the 3 phyla. Our results suggest that the fiber level and growth stages could impact the composition of gut microbiota.


Asunto(s)
Bacterias/metabolismo , Fibras de la Dieta/metabolismo , Digestión , Metabolismo Energético , Gansos/microbiología , Gansos/fisiología , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales/efectos de los fármacos , Animales , Bacterias/efectos de los fármacos , Dieta/veterinaria , Fibras de la Dieta/administración & dosificación , Digestión/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Metabolismo Energético/efectos de los fármacos , Femenino , Microbioma Gastrointestinal , Masculino , Distribución Aleatoria
11.
Medicine (Baltimore) ; 97(23): e10969, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29879048

RESUMEN

BACKGROUND: Myoclonus is an undesirable phenomenon that occurs after induction of general anesthesia using etomidate. Opioids such as sufentanil are considered effective pretreatment drugs for myoclonus inhibition, although high doses are required. Transcutaneous acupoint electrical stimulation (TAES), a noninvasive technique involving electrical stimulation of the skin at the acupuncture points, exhibits analgesic effects, promotes anesthetic effects, decreases the dose of anesthetic drugs, and increases endogenous opioid peptide levels. In the present study, we investigated the effects of TAES combined with low-dose sufentanil pretreatment on the incidence and severity of etomidate-induced myoclonus in patients undergoing elective hysteroscopy. METHODS: In a double-blind manner, 172 patients (American Society of Anesthesiologists class I-II; age, 20-55 years) scheduled to undergo elective hysteroscopy were randomized into the following groups (n = 43 each): control (false TAES followed by saline injection after 30 min), TAES (TAES followed by saline injection after 30 minutes), sufentanil [false TAES followed by low-dose sufentanil (0.1 µg/kg) injection after 30 minutes], and sufentanil plus TAES (TAES followed by low-dose sufentanil injection after 30 minutes). In all groups, general anesthesia was induced by etomidate 0.3 mg/kg after sufentanil or saline injection. The incidence and severity of myoclonus were assessed for 2 minutes after etomidate administration. The visual analogue scale (VAS) scores for pain at 1 hour after surgery were recorded. The heart rate (HR), mean arterial pressure (MAP), and peripheral capillary oxygen saturation (SPO2) were recorded before premedication, after etomidate injection, after uterus expansion, and after recovery from anesthesia. RESULTS: The incidence of myoclonus was highest in the control group (88.3%), followed by TAES (74.4%), sufentanil (60.4%), and TAES plus sufentanil (48.8%) groups. Thus, the incidence was significantly higher in the control and TAES groups than in the sufentanil and TAES plus sufentanil groups. Grade 3 myoclonus occurred in 30.2%, 9.3%, 11.6%, and 9.3% patients in the control, TAES, sufentanil, and TAES plus sufentanil groups, respectively, with significant differences between the control group and the other 3 groups. Furthermore, the postoperative VAS scores for pain were significantly lower in the TAES, sufentanil, and TAES plus sufentanil groups compared with those in the control group. There were no significant differences in any other parameters among groups. CONCLUSION: Our results suggest that TAES combined with low-dose opioids such as sufentanil can decrease the incidence and severity of etomidate-induced myoclonus.


Asunto(s)
Anestésicos Intravenosos/administración & dosificación , Etomidato/efectos adversos , Mioclonía/prevención & control , Sufentanilo/administración & dosificación , Estimulación Eléctrica Transcutánea del Nervio/métodos , Puntos de Acupuntura , Adulto , Anestésicos Intravenosos/efectos adversos , Terapia Combinada , Método Doble Ciego , Femenino , Humanos , Histeroscopía/efectos adversos , Histeroscopía/métodos , Incidencia , Persona de Mediana Edad , Mioclonía/inducido químicamente , Mioclonía/epidemiología , Resultado del Tratamiento , Adulto Joven
12.
Theriogenology ; 96: 103-110, 2017 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-28532825

RESUMEN

Genetic modification provides a means to enhancing disease resistance in animals. Toll-like receptor 4 (TLR4), a member of the TLR family, is critical for the recognition of lipopolysaccharide (LPS)/endotoxin from Gram-negative bacteria by host immune cells, which initiates cell activation and subsequently triggers a proinflammatory response to the invading pathogens. In this study, the first generation of genetically modified (GM) sheep overexpressing TLR4 was produced by microinjection for better disease resistance. Compared with wild-type (WT) rams, the GM rams have similar growth performance, basic semen quality and spermatozoon ultrastructure. The offspring birth rates after cervical artificial insemination were also similar between GM (90.32%) and WT (92.38%) rams. Overall, the presence and expression of the TLR4 transgene in the genome did not appear to interfere with normal semen production, reproductive traits and the ability of transgene transmission to offspring. The expression levels of TLR4, tumor necrosis factor and interferon gamma genes in monocyte/macrophages from GM sheep were significantly higher than that from WT sheep at early stages after LPS stimulation. The GM offspring born from the founder transgenic ram inseminated ewes had similar survival rate with WT offspring (88.89% vs 84.86%) at weaning. The TLR4 transgene showed no deleterious effects on growth performance, reproductive traits and offspring survivability of GM rams. Therefore, the GM sheep overexpressing TLR4 provide a powerful experimental model for analyzing function of TLR4 in vivo during infection and inflammation.


Asunto(s)
Animales Modificados Genéticamente , Ovinos/genética , Ovinos/fisiología , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Animales , Longevidad , Preservación de Semen
13.
BMC Vet Res ; 8: 196, 2012 Oct 22.
Artículo en Inglés | MEDLINE | ID: mdl-23082910

RESUMEN

BACKGROUND: Toll-like receptor 2 (TLR2) is important to host recognition of invading gram-positive microbes. In goats, these microbes can cause serious mastitis, anthrax, tetanus, and other problems. Transgenic goats constitutively over-expressing TLR2 in many tissues serve as a suitable model for the study of the role of TLR2 over-expression in bacterial clearance. RESULTS: Capra hircus TLR2 over-expression vector (p3S-LoxP-TLR2) was used to generate transgenic goats by egg microinjection. The integration efficiency was 8.57%. Real-time PCR and immunohistochemical results confirmed that the goats over-expressing the TLR2 gene (Tg) expressed more TLR2 than wild-type goats (WT). Monocyte-macrophages from the bloodstreams of transgenic goats were stimulated with synthetic bacterial lipoprotein (Pam3CSK4) and by the promotion of interleukin-6 (IL-6) and IL-10 expression in vitro. The oxidative damage was significantly reduced, and lysozyme (LZM) secretion was found to be up-regulated. Ear tissue samples from transgenic goats that had been stimulated with Pam3CSK4 via hypodermic injection showed that transgenic individuals can undergo the inflammation response very quickly. CONCLUSIONS: Over-expression of TLR2 was found to decrease radical damage to host cells through low-level production of NO and MDA and to promote the clearance of invasive bacteria by up-regulating lysozyme secretion and filtration of inflammatory cells to the infected site.


Asunto(s)
Enfermedades de las Cabras/metabolismo , Muramidasa/metabolismo , Estrés Oxidativo/fisiología , Receptor Toll-Like 2/biosíntesis , Animales , Animales Modificados Genéticamente , Femenino , Enfermedades de las Cabras/genética , Enfermedades de las Cabras/inmunología , Cabras , Histocitoquímica/veterinaria , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Lipopolisacáridos/farmacología , Malondialdehído/metabolismo , Muramidasa/inmunología , Óxido Nítrico/metabolismo , ARN/química , ARN/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/inmunología , Regulación hacia Arriba
14.
Chin Med Sci J ; 24(2): 86-90, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19618604

RESUMEN

OBJECTIVE: To evaluate the significance of several Dermatophagoides pteronyssinus allergen extracts for skin prick test (SPT) in patients allergic to Dermatophagoides pteronyssinus. METHODS: Two hundred and nineteen patients enrolled in Peking Union Medical College Hospital underwent SPT and serum specific IgE assay to detect the Dermatophagoides pteronyssinus allergen. Three kinds of house dust mite allergen extracts were used for SPT, including the Dermatophagoides pteronyssinus extract prepared by our laboratory (group A), standardized Dermatophagoides pteronyssinus extract (group B), and mixed extracts of Dermatophagoides pteronyssinus and Dermatophagoides farinae (group C). Human serum specific IgE result was regarded as the reference standard for diagnosis of Dermatophagoides pteronyssinus allergy. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic performance of SPT with the extracts of three groups. RESULTS: SPT results showed that the median wheal diameter of group A, group B, and group C was 0.43, 0.35, and 0.28 cm, respectively, with significant difference among three groups (P<0.05). The difference was significant between group A and B (P<0.01) as well as group A and C (P<0.01), but not between group B and C (P>0.05). There was no local urticaria or systemic allergic reactions following the procedure of SPT. Local reaction was observed in 5 patients and delayed reaction was in 2 patients of group A. As for group B and C, local reaction occurred in 3 cases and delayed reaction in 2 cases in each group. The area under ROC curve of SPT with extract in group A, group B, and group C was 0.765, 0.801, and 0.782, respectively. Based on the detection results of serum specific IgE, the sensitivity of SPT in diagnosis of Dermatophagoides pteronyssinus allergy with extract of group A, group B, and group C was 92.4%, 87.0%, and 81.5%, and the specificity was 60.6%, 73.2%, and 74.8%, respectively. CONCLUSION: The Dermatophagoides pteronyssinus extract for SPT prepared by our laboratory offers good sensitivity and specificity comparable to commercially available allergen extracts, and it may be an appropriate candidate for clinical screening and diagnosis of Dermatophagoides pteronyssinus allergy.


Asunto(s)
Antígenos Dermatofagoides , Dermatophagoides pteronyssinus/inmunología , Pruebas Cutáneas/métodos , Animales , Antígenos Dermatofagoides/inmunología , Femenino , Humanos , Masculino , Curva ROC , Sensibilidad y Especificidad
15.
Zhonghua Yi Xue Za Zhi ; 86(25): 1759-63, 2006 Jul 04.
Artículo en Chino | MEDLINE | ID: mdl-17054843

RESUMEN

OBJECTIVE: To evaluate the value of intradermal skin test (IDT) and serum sIgE detection in diagnosing Artemisia sensitivity in Chinese patients with autumnal hay fever. METHODS: 1150 patients with autumnal rhinitis or asthma, 504 males and 646 females, aged 5 approximately 77, were evaluated by experienced physicians, then underwent IDT by using 20 kinds of aeroallergen extracts. The concentrations of Artemisia and Ragweed extracts employed in skin test were 1:1000 (W/V) and the concentrations of other aeroallergens were all 1:100 (W/V). Then all patients underwent detection of Artemisia sIgE. Diagnostic standards were established based on the results of IDT and sIgE results respectively. A reference standard was established according to the typical history, symptoms, and an wheal with a diameter >or= 5mm and a sIgE level >or= 0.35 kU(A)/L, an wheal with the diameter >or= 10 mm alone; or a sIgE level >or= 0.70 kUa/L alone. RESULTS: When using the reference standard as criteria, using IDT had better sensitivity (96.2%), specificity (74.2%), positive predictive value (+PV, 93.5%), negative predictive value (-PV, 85.7%), and efficiency (91.6%) than using sIgE >or= 0.35 kUa/L alone as the criteria of IDT; sIgE detection had better sensitivity (97.6%), specificity (94.9%), +PV (98.7%), -PV (91.1%), and efficiency (97.0%) than using wheal diameter >or= 5 mm alone as the criteria of sIgE detection. The false positive rate of IDT and sIgE detection decreased from 35% and 22.7% to 25.6% and 5.1% respectively when using the wheal diameter >or= 10 mm or sIgE >or= 0.70 kUa/L as a positive criteria. CONCLUSION: IDT and sIgE detection are correlated with each other well in diagnosing Artemisia pollinosis, both of them have the possibility of being false positive, but IDT has higher false positive rate than sIgE detection. The false positive rates of IDT and sIgE detection can be decreased by increasing the positive criteria to higher grading reaction.


Asunto(s)
Alérgenos/sangre , Artemisia , Inmunoglobulina E/sangre , Rinitis Alérgica Estacional/diagnóstico , Adolescente , Adulto , Anciano , Artemisia/inmunología , Asma/diagnóstico , Niño , Preescolar , Femenino , Humanos , Pruebas Intradérmicas/normas , Masculino , Persona de Mediana Edad
16.
Zhonghua Yi Xue Za Zhi ; 86(27): 1906-11, 2006 Jul 18.
Artículo en Chino | MEDLINE | ID: mdl-17064529

RESUMEN

OBJECTIVE: To evaluate the values of intradermal skin test (IDT) and serum specific IgE detection in diagnosing Humulus scandens sensitivity in Chinese patients with autumnal hay fever. METHODS: 1150 patients, 504 males and 646 females, aged 5 approximately 75, were evaluated as with autumnal rhinitis and asthma by experienced physicians. Then the patients underwent IDT by using 20 kinds of aeroallergen extracts, at the concentration of 1:1000 (w/v) for Artemisia spp. and ragweed or at the concentration of 1:100 (w/v) for Humulus scandens and others. The Humulus sIgE level was performed in all patients. A diagnosing criteria was established according to typical history, symptoms, and any one of the following findings: (1) wheal > or = 5 mm in diameter by IDT and sIgE to Humulus of the concentration > or = 0.35 kUa/L at same time; (2) wheal > or = 10 mm in diameter by IDT alone; and (3) sIgE to Humulus of the concentration > or = 0.70 kUa/L alone. RESULTS: When using the above diagnosing criteria as the reference standard, IDT had better sensitivity (97.2%), positive predictive value (PV+) (77.9%), negative predictive value (PV-) (90.6%) and efficiency (80.4% than using sIgE > or = 0.35 kUa/L alone as the reference criteria of IDT, however, had a lower specificity (49.8%); and sIgE detection had better sensitivity (89.0%), specificity (97.5%), PV+ (98.8%), PV- (88.0%) and efficiency (92.0%) than using wheal > or = 5 mm in diameter alone as the reference criteria of sIgE detection. The false positive rate of IDT decreased from 59.9% to 17.4% when using wheal diameter > or = 10 mm as the positive criteria. In the 288 patients with a negative sIgE result and a positive IDT result 84 cases had stronger positive IDT result (with the wheal diameter > or = 10 mm). CONCLUSION: IDT is correlated well with sIgE detection in diagnosing Humulus scandens pollinosis, but the false positive rate of IDT is higher than that of sIgE test. The false positive rate of IDT can be decreased by increasing the positive criteria to higher grading reaction. The immunotherapy according to the lower grading positive result of IDT alone or sIgE alone should be avoided.


Asunto(s)
Alérgenos/análisis , Humulus/inmunología , Inmunoglobulina E/sangre , Rinitis Alérgica Estacional/diagnóstico , Adolescente , Adulto , Anciano , Asma/diagnóstico , Asma/epidemiología , Niño , Preescolar , China/epidemiología , Femenino , Humanos , Pruebas Intradérmicas , Masculino , Persona de Mediana Edad , Rinitis Alérgica Estacional/epidemiología
17.
Zhonghua Yi Xue Za Zhi ; 86(23): 1628-32, 2006 Jun 20.
Artículo en Chino | MEDLINE | ID: mdl-16854304

RESUMEN

OBJECTIVE: To investigate the natural course from rhinitis to asthma in the patients with autumnal pollinosis. METHODS: 1096 patients with autumnal pollinosis, 488 males and 632 females, aged 38 +/- 14 (5 - 77), 511 with pure allergic rhinitis and 585 with allergic rhinitis complicated with asthma, underwent skin test of inhalant allergens, serum IgE specific to autumnal pollens, and questionnaire survey. RESULTS: The age range of rhinitis induced by autumnal pollens was 2 - 75. The peak onset age of rhinitis was 15 - 44, and the peak onset age of asthma was 25 - 54. 33% of the rhinitis patients complicated with asthma (194/585) had their first attack of rhinitis and the first attack of asthma in the same year, 66% of them (386/585) had the first attack of asthma later than the first attack of rhinitis, and only 0.8% of them (5/585) had their first attack of asthma earlier than that of rhinitis. 37% of the patients with autumnal pollen allergic rhinitis (410/1096) developed asthma within 5 years, 47% (511/1096), within 9 years, 5% (58/1096) within 10 approximately 19 years, and 1.5% (16/1096) within 20 - 40 years. CONCLUSION: Almost half of the patients with autumnal pollen allergic rhinitis develop to seasonal allergic asthma within 9 years.


Asunto(s)
Asma/epidemiología , Rinitis Alérgica Estacional/epidemiología , Adolescente , Adulto , Edad de Inicio , Anciano , Asma/diagnóstico , Niño , Preescolar , Femenino , Humanos , Inmunoglobulina E/sangre , Pruebas Intradérmicas , Masculino , Persona de Mediana Edad , Rinitis Alérgica Estacional/diagnóstico , Encuestas y Cuestionarios
18.
Zhonghua Yi Xue Za Zhi ; 85(24): 1683-7, 2005 Jun 29.
Artículo en Chino | MEDLINE | ID: mdl-16251071

RESUMEN

OBJECTIVE: To investigate the relationship between allergic rhinitis and allergic asthma in the patients with autumnal pollinosis. METHODS: 1120 patients with autumnal pollinosis, aged 5 - 75, excluding those with typical symptoms of seasonal rhinitis or asthma but with positive skin test and serum IgE specific to dustmite and fungi, underwent standardized clinical questionnaire survey, including the onset age, onset time, and symptoms as well as the severity of asthma, skin tests, and examination of serum IgE specific to autumnal pollens. RESULTS: The average onset age of the allergic rhinitis patients induced by autumnal pollens was 27.9 years, significantly younger than that of the allergic asthma patients (32.6 years, P < 0.001). Out of the 1120 patients 1096 (97.9%) had allergic rhinitis, 602 (53.8%) had asthma, 507 (45.3%) had allergic rhinitis only, and 10 (0.9%) had allergic asthma only. Among the 1096 patients with allergic rhinitis 585 (53.4%) suffered from seasonal asthma. Among the 602 patients with asthma 585 (97.2%) suffered from seasonal rhinitis, and 183 of the 602 patients (30.8%) needed emergency treatment. CONCLUSION: Autumnal pollens are very important causes which induce asthma during autumnal season in northern China.


Asunto(s)
Asma/complicaciones , Rinitis Alérgica Perenne/complicaciones , Rinitis Alérgica Estacional/complicaciones , Adolescente , Adulto , Factores de Edad , Anciano , Asma/epidemiología , Niño , Preescolar , China/epidemiología , Femenino , Humanos , Inmunoglobulina E/sangre , Masculino , Persona de Mediana Edad , Rinitis Alérgica Perenne/epidemiología , Rinitis Alérgica Estacional/sangre , Rinitis Alérgica Estacional/epidemiología
19.
Chin Med Sci J ; 20(1): 27-9, 2005 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-15844308

RESUMEN

OBJECTIVE: To explore the effects of SDS, PBS re-dissolvent solutions on fluorescence values of radioallergosorbent test (RAST) inhibition. METHODS: Dermatophagoides pterronyssinus allergen immunoCAP and UniCAP 100 System were used. The Sera Pool consisted of 20 Dermatophagoides pterronyssinus allergic patients sera, their specific IgE fluorescence values were between 12505 and 24776. RESULTS: Fluorescence value percentages decreased: 62.9%, 54.1%, 43.5%, 6.7%, 3.7%, 2.6%, 2.2%, and 1.4% respectively, when SDS concentrations were at 2%, 1%, 0.5%, 0.25%, 0.1%, 0.05%, 0.025%, and 0.01%. Fluorescence values decreased more than 5% with SDS concentrations equal to 0.25% or higher. PBS in 0.1 and 0.01 mol/L concentrations decreased fluorescence values 2.9% and 0.9% respectively. CONCLUSIONS: SDS is a commonly used surfactants in allergen extract and re-dissolvent prepared allergen precipitation for RAST inhibition. Thus effects of surfactants (e.g. SDS) upon the RAST inhibition tests must be considered when they were used as re-dissolvent agents to improve protein resolution in RAST inhibition.


Asunto(s)
Dermatophagoides pteronyssinus/inmunología , Fosfatos/administración & dosificación , Prueba de Radioalergoadsorción/métodos , Dodecil Sulfato de Sodio/administración & dosificación , Tensoactivos/administración & dosificación , Alérgenos , Relación Dosis-Respuesta a Droga , Fluorescencia , Humanos , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Fosfatos/farmacología , Dodecil Sulfato de Sodio/farmacología , Soluciones , Tensoactivos/farmacología
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