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Plant-derived ß-glucosidases hold promise for glycoside biosynthesis via reverse hydrolysis because of their excellent glucose tolerance and robust stability. However, their poor heterologous expression hinders the development of large-scale production and applications. In this study, we overexpressed apple seed ß-glucosidase (ASG II) in Komagataella phaffii and enhanced its production from 289 to 4322 U L-1 through expression cassette engineering and protein engineering. Upon scaling up to a 5-L high cell-density fermentation, the resultant mutant ASG IIV80A achieved a maximum protein concentration and activity in the secreted supernatant of 2.3 g L-1 and 41.4 kU L-1, respectively. The preparative biosynthesis of salidroside by ASG IIV80A exhibited a high space-time yield of 33.1 g L-1 d-1, which is so far the highest level by plant-derived ß-glucosidase. Our work addresses the long-standing challenge of the heterologous expression of plant-derived ß-glucosidase in microorganisms and presents new avenues for the efficient production of salidroside and other natural glycosides.
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Glucósidos , Malus , Fenoles , Semillas , beta-Glucosidasa , Fenoles/metabolismo , beta-Glucosidasa/genética , beta-Glucosidasa/metabolismo , Glucósidos/biosíntesis , Glucósidos/metabolismo , Glucósidos/química , Semillas/genética , Semillas/metabolismo , Saccharomycetales/genética , Saccharomycetales/metabolismo , Saccharomycetales/enzimología , Fermentación , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ingeniería de Proteínas/métodosRESUMEN
BACKGROUND: Pathological axillary lymph node (pALN) burden is an important factor for treatment decision-making in clinical T1-T2 (cT1-T2) stage breast cancer. Preoperative assessment of the pALN burden and prognosis aids in the individualized selection of therapeutic approaches. PURPOSE: To develop and validate a machine learning (ML) model based on clinicopathological and MRI characteristics for assessing pALN burden and survival in patients with cT1-T2 stage breast cancer. STUDY TYPE: Retrospective. POPULATION: A total of 506 females (range: 24-83 years) with cT1-T2 stage breast cancer from two institutions, forming the training (N = 340), internal validation (N = 85), and external validation cohorts (N = 81), respectively. FIELD STRENGTH/SEQUENCE: This study used 1.5-T, axial fat-suppressed T2-weighted turbo spin-echo sequence and axial three-dimensional dynamic contrast-enhanced fat-suppressed T1-weighted gradient echo sequence. ASSESSMENT: Four ML methods (eXtreme Gradient Boosting [XGBoost], Support Vector Machine, k-Nearest Neighbor, Classification and Regression Tree) were employed to develop models based on clinicopathological and MRI characteristics. The performance of these models was evaluated by their discriminative ability. The best-performing model was further analyzed to establish interpretability and used to calculate the pALN score. The relationships between the pALN score and disease-free survival (DFS) were examined. STATISTICAL TESTS: Chi-squared test, Fisher's exact test, univariable logistic regression, area under the curve (AUC), Delong test, net reclassification improvement, integrated discrimination improvement, Hosmer-Lemeshow test, log-rank, Cox regression analyses, and intraclass correlation coefficient were performed. A P-value <0.05 was considered statistically significant. RESULTS: The XGB II model, developed based on the XGBoost algorithm, outperformed the other models with AUCs of 0.805, 0.803, and 0.818 in the three cohorts. The Shapley additive explanation plot indicated that the top variable in the XGB II model was the Node Reporting and Data System score. In multivariable Cox regression analysis, the pALN score was significantly associated with DFS (hazard ratio: 4.013, 95% confidence interval: 1.059-15.207). DATA CONCLUSION: The XGB II model may allow to evaluate pALN burden and could provide prognostic information in cT1-T2 stage breast cancer patients. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY: Stage 2.
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Enamel is composed of numerous uniformly wide, well-oriented hydroxyapatite crystals. It possesses an acellular structure that cannot be repaired after undergoing damage. Therefore, remineralization after enamel defects has become a focal point of research. Hydrogels, which are materials with three-dimensional structures derived from cross-linking polymers, have garnered significant attention in recent studies. Their exceptional properties make them valuable in the application of enamel remineralization. In this review, we summarize the structure and formation of enamel, present the design considerations of hydrogels for enamel remineralization, explore diverse hydrogels types in this context, and finally, shed light on the potential future applications in this field.
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BACKGROUND: Although radical surgical resection is the most effective treatment for hepatocellular carcinoma (HCC), the high rate of postoperative recurrence remains a major challenge, especially in patients with alpha-fetoprotein (AFP)-negative HCC who lack effective biomarkers for postoperative recurrence surveillance. Emerging radiomics can reveal subtle structural changes in tumors by analyzing preoperative contrast-enhanced computer tomography (CECT) imaging data and may provide new ways to predict early recurrence (recurrence within 2 years) in AFP-negative HCC. In this study, we propose to develop a radiomics model based on preoperative CECT to predict the risk of early recurrence after surgery in AFP-negative HCC. PATIENTS AND METHODS: Patients with AFP-negative HCC who underwent radical resection were included in this study. A computerized tool was used to extract radiomic features from the tumor region of interest (ROI), select the best radiographic features associated with patient's postoperative recurrence, and use them to construct the radiomics score (RadScore), which was then combined with clinical and follow-up information to comprehensively evaluate the reliability of the model. RESULTS: A total of 148 patients with AFP-negative HCC were enrolled in this study, and 1,977 radiographic features were extracted from CECT, 2 of which were the features most associated with recurrence in AFP-negative HCC. They had good predictive ability in both the training and validation cohorts, with an area under the ROC curve (AUC) of 0.709 and 0.764, respectively. Tumor number, microvascular invasion (MVI), AGPR and radiomic features were independent risk factors for early postoperative recurrence in patients with AFP-negative HCC. The AUCs of the integrated model in the training and validation cohorts were 0.793 and 0.791, respectively. The integrated model possessed the clinical value of predicting early postoperative recurrence in patients with AFP-negative HCC according to decision curve analysis, which allowed the classification of patients into subgroups of high-risk and low-risk for early recurrence. CONCLUSION: The nomogram constructed by combining clinical and imaging features has favorable performance in predicting the probability of early postoperative recurrence in AFP-negative HCC patients, which can help optimize the therapeutic decision-making and prognostic assessment of AFP-negative HCC patients.
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Carcinoma Hepatocelular , Medios de Contraste , Neoplasias Hepáticas , Recurrencia Local de Neoplasia , Tomografía Computarizada por Rayos X , alfa-Fetoproteínas , Humanos , Carcinoma Hepatocelular/cirugía , Carcinoma Hepatocelular/diagnóstico por imagen , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/cirugía , Neoplasias Hepáticas/diagnóstico por imagen , Neoplasias Hepáticas/patología , Masculino , Femenino , alfa-Fetoproteínas/metabolismo , alfa-Fetoproteínas/análisis , Recurrencia Local de Neoplasia/diagnóstico por imagen , Persona de Mediana Edad , Tomografía Computarizada por Rayos X/métodos , Anciano , Estudios Retrospectivos , Adulto , Hepatectomía , Pronóstico , RadiómicaRESUMEN
OBJECTIVE: This study aims to investigate the suicide risk and mental health status of non-psychiatric inpatients in general hospital and explore the risk factors of suicide. METHODS: A prospective study was conducted at a tertiary general hospital in Guangzhou, Guangdong Province, China. On-line assessment of mental health status and suicide was completed at admission and discharge. We assessed depression, anxiety, insomnia and suicide of inpatients and binary logistics regression was used to examine the risk factors of suicide. RESULTS: From April 1, 2021 and January 31, 2022, 3685 inpatients were included. The detection rates of depression, anxiety and insomnia were 14.6 %, 9.0 % and 17.8 %, respectively. There were 2.7 % of inpatients at suicide risk. Binary logistics regression demonstrated that the inpatients with anxiety were at higher risk of suicide. LIMITATIONS: (1) Single-center study limits the generalization of conclusion, (2) low response rate at discharge. CONCLUSIONS: The comorbidity of physical illnesses and mental health problems, including depression, anxiety, insomnia, and suicide among non-psychiatric patients in general hospital was common. An assessment of anxiety may help identify individuals at high suicide risk. Medical staff in general hospitals should be trained to improve their ability to identify mental disorders and high-risk individuals for suicide, provide timely interventions and effectively reduce the suicide risk of patients.
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Trastornos Mentales , Trastornos del Inicio y del Mantenimiento del Sueño , Suicidio , Humanos , Hospitales Generales , Trastornos del Inicio y del Mantenimiento del Sueño/epidemiología , Pacientes Internos/psicología , Prevalencia , Estudios Prospectivos , Trastornos Mentales/epidemiología , Trastornos Mentales/psicología , Suicidio/psicología , Factores de RiesgoRESUMEN
Qingtian paddy field carp (PF-carp) is a local carp cultivated in the paddy field of Qingtian, Zhejiang. This rice-fish co-culture system has been recognized as one of the Globally Important Agriculture Heritage Systems (GIAHS). PF-carp has been acclimatized to the high-temperature environment of shallow paddy fields after several centuries of domestication. To reveal the physiological and molecular regulatory mechanisms of PF-carp, we chose to use 28°C as the control group and 34°C as the treatment group. We measured biochemical parameters in their serum and hepatopancreases and also performed transcriptome sequencing analysis. Compared with the control group, the serum levels of malondialdehyde (MDA), glucose (GLU), glutathione peroxidase (GSH-Px), catalase (CAT), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) show no significant change. In addition, superoxide dismutase (SOD), GSH-Px, and CAT also show no significant change in hepatopancreases. We identified 1,253 differentially expressed genes (DEGs), and their pathway analysis revealed that heat stress affected AMPK signaling pathway, protein export, and other biological processes. It is worth noting that protein processing in the endoplasmic reticulum (ER) was the most significantly enriched pathway identified by the Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene set enrichment analysis (GSEA). Significantly higher levels of HSP40, HSP70, HSP90, and other ubiquitin ligase-related genes were upregulated. In summary, heat stress did not lead to tissue damage, inflammation, oxidative stress, and ER stress in the hepatopancreases of PF-carp. This study provides valuable insights into the adaptation mechanism of this species to the high-temperature environment of paddy fields.
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OBJECTIVE: To identify the difference in clinical characteristics between viral pneumonia and Mycoplasma pneumoniae , providing cues on their differential diagnosis for primary hospitals with the insufficient pathogen detection capacity. METHODS: We retrospectively reviewed the medical records of hospitalized children with acute respiratory tract infections, and pathogenic microbes test results were analyzed. Clinical characteristics, routine blood parameters and hospitalization duration and fee were compared between M. pneumoniae and viral pneumonia. We used in the multivariable logistic regression to predict the probability of children with M. pneumoniae and graphically represented by a dynamic nomogram. The discrimination and clinical utility of the model were confirmed by receiver operating characteristic and decision curve analysis curves. RESULT: A total of 375 children with community-acquired pneumonia were included. Mycoplasma infection accounted for the largest proportion (22.13%). The incidence of both hypothermia and vomiting was lower in M. pneumoniae compared to viral pneumonia (hypothermia: 10.50% vs. 0.00%; vomiting: 7.90% vs. 0.00%). The prevalence of hyperthermia was higher in M. pneumoniae (hyperthermia: 89.5% vs. 100%). Procalcitonin, peripheral blood white blood cell count and lymphocyte levels were higher in the viral pneumonia group, and eosinophil levels were conversely lower. As for the duration of illness, the mean length of stay was 5.20 ± 2.12 (viral pneumonia) and 6.27 ± 2.48 days ( M. pneumoniae ). Children with M. pneumoniae had higher overall hospital costs and required more medical treatment. The above were all statistically significant with a P < 0.05. The scoring system was established based on the above results. Receiver operating characteristic curves showed good model-discrimination ability with 0.844 of the area under the curve in the training set and 0.778 in the test set. Decision curve analysis curves demonstrated the discriminative superiority of this model. The web-based dynamic nomogram calculator is accessible at https://zhxylxy0160128.shinyapps.io/Nomogram/ . CONCLUSION: Nomograms have satisfactory discrimination, and clinical utility may benefit in predicting the probability of developing M. pneumoniae in children. Children with M. pneumoniae have a higher burden than those with viral pneumonia and may require more intensive in-hospital monitoring.
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Infecciones Comunitarias Adquiridas , Hipotermia , Neumonía por Mycoplasma , Neumonía Viral , Niño , Humanos , Mycoplasma pneumoniae , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/epidemiología , Neumonía por Mycoplasma/tratamiento farmacológico , Estudios Retrospectivos , Nomogramas , Hospitales , Vómitos , Infecciones Comunitarias Adquiridas/diagnóstico , Infecciones Comunitarias Adquiridas/epidemiologíaRESUMEN
Trigeminal neuralgia (TN) is a common facial neuropathic pain that is mainly characterized by spontaneous or induced needling or electric shock pain in the innervation area of the trigeminal nerve. It is also referred to as "the cancer that never dies". The olfactory ensheathing cell (OEC) is a special glial cell in the nervous system that has a strong supportive function in nerve regeneration. Cell transplantation therapy is a useful treatment modality that we believe can be applied in TN management. In this study, OECs were transplanted into the ligation site of the infraorbital nerve of rats. We found that after the OEC transplantation, mechanical pain threshold in the face of the rats was significantly increased. Western blotting, immunofluorescence assay, and reverse transcription-quantitative polymerase chain reaction were performed on the trigeminal ganglia (TG) of model rats. The results revealed a decrease in the expression of P2X7 receptor (P2X7R) in the trigeminal ganglia. Our findings show that OEC transplantation has a good therapeutic effect on TN in rats, and that can reduce the expression of P2X7R in trigeminal ganglia. Therefore, we think that OEC transplantation may be a suitable treatment for TN.
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Schwann cells are components of the peripheral nerve myelin sheath, which supports and nourishes axons. Upon injury of the trigeminal nerve, Schwann cells are activated and cause trigeminal neuralgia by engulfing the myelin sheath and secreting various neurotrophic factors. Further, Schwann cells can repair the damaged nerve and thus alleviate trigeminal neuralgia. Here, we briefly describe the development and activation of Schwann cells after nerve injury. Moreover, we expound on the occurrence, regulation, and treatment of trigeminal neuralgia; further, we point out the current research deficiencies and future research directions.
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Células de Schwann/patología , Neuralgia del Trigémino/patología , Animales , Humanos , Neuralgia del Trigémino/terapiaRESUMEN
The presence of sufficient dissolved oxygen (DO) in a constructed wetland (CW) is vital to the process of removing ammonia nitrogen and organics from wastewater. To achieve total nitrogen removal, which is characterised by enhanced ammonia nitrogen removal, this study offers an efficient strategy to increase the oxygen supply by establishing constant unsaturated zones and baffles in simulating constructed wetlands (SCWs). Henceforth, this strategy is addressed as a partially unsaturated SCW. A centrally located high tube was set up inside the wetland to create an unsaturated zone at a higher level. The effectiveness of the unsaturated zone to supplement the oxygen content was evaluated by comparing with controls (an unaerated SCW and an aerated SCW). The results show the chemical oxygen demand removal rate (85⯱â¯6%) in the partially unsaturated SCW was equivalent to that in the aerated SCW (83⯱â¯6%), while the ammonia nitrogen removal rate was 11 times higher compared to that of the unaerated SCW. The removal potential of the partially unsaturated SCW under different HRT (hydraulic retention time)s (12, 24, and 36â¯h) was examined, and the 36â¯h-SCW performed the best in the removal of organics and nitrogen. The mechanisms behind the unsaturated zone strategy were studied by analysing water and microbe samples along the pathway. The results from the water quality indicators and the quantitative polymerase chain reactions along the pathway showed the unsaturated zone contributed to the removal of primary organics and ammonia nitrogen. The superior performance of unsaturated zone strategy was discussed further using the enrichment of ammonia-oxidising bacteria, mass of oxygen uptake, and baffle design. The results indicate that the amoA gene/16s rRNA gene abundance ratio and the oxygen uptake (336⯱â¯44â¯gâ¯m-3 d-1) in the partially unsaturated SCW was higher than that observed in the two controls.
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Eliminación de Residuos Líquidos/métodos , Humedales , Amoníaco , Análisis de la Demanda Biológica de Oxígeno , Nitrógeno , Aguas ResidualesRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: San Wei Gan jiang San (SWGJS) also called Jia Ga Song Tang, is widely used in ancient medicine for liver diseases. THE AIM OF THIS STUDY: To identify the blood components of SWGJS. To determine the hepatoprotective effect and the mechanism of SWGJS by observing its effect on different degrees of liver damage and gene knockdown cells. MATERIALS AND METHODS: SWGJS treated serum was analyzed by UPLC-MS to identify blood components. CCl4-induced chronic liver injury in rats was treated with SWGJS. The viscera index was calculated. Pathological changes of the liver were determined by HE staining and analysis of by following: GSH-Px and MDA in liver homogenate; ALT and AST in serum; mRNA expression of Nrf2, Bach1, and HO-1 by RT-PCR; Nrf2 and Bach1 in the nucleus and cytoplasm; HO-1 total expression by Western blot; silencing Nrf2 and Bach1 in human L-02 cells by siRNA; MDA, GSH-Px, GST, and GR in cell supernatants; and GSH/GSSG within the cell. RESULTS: We found that 6-gingerol was one of the blood components in the serum treated with SWGJS. In CCl4-induced chronic liver injury in rats, SWGJS repaired the liver structure in the early stages of liver damage as evidenced by reduced ALT and AST in the serum, increased GSH-Px activity and decreased MDA levels in the liver over time. SWGJS has excellent antioxidant and hepatoprotective effects and prevents disease progression. The mechanism of SWGJS is related to the dynamics promoting Nrf2 entry to the nucleus and Bach1 exit from the nucleus. In L-02 cells with silenced Nrf2, the antioxidant enzyme system was disordered, and the change in the cellular redox state was not conducive to antioxidative stress. However, in cells with silenced Bach1, the antioxidant enzyme system could be activated to promote cellular antioxidant stress. SWGJS had a combined effect on Nrf2 and Bach1 contributing to antioxidant properties and liver protection. SWGJS increased GSH-Px and HO-1, decreased MDA and increased the ratio of GSH/GSSG by upregulating the expression of Nrf2 to enhance its antioxidant effects. At the same time, SWGJS had a specific impact on decreasing Bach1. Its elevation of GST is due to the overall performance of increasing Nrf2 and decreasing Bach1. This mechanism of action embodies the characteristics of the multitarget impact of traditional medicine and the antioxidation effect of SWGJS. CONCLUSIONS: 6-Gingerol is one of the blood components of SWGJS. SWGJS can regulate antioxidant enzymes, protect against liver damage in different stages, and slow the progression of liver cell damage and liver disease by increasing Nrf2 and reducing Bach1 in the nucleus, dynamically regulating Nrf2/Bach1.
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Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Animales , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Tetracloruro de Carbono , Enfermedad Hepática Inducida por Sustancias y Drogas/genética , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Hemo Oxigenasa (Desciclizante)/genética , Hemo Oxigenasa (Desciclizante)/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Masculino , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To study the effect of montelukast sodium (MK), a leukotriene receptor antagonist, and bacterial lysates (OM-85BV), used alone or in combination, on airway remodeling and the expression of transforming growth factor-ß1 (TGF-ß1) and Smad7 in guinea pigs with bronchial asthma and their correlation. METHODS: A total of 40 male Hartley guinea pigs were randomly divided into normal control group, asthma group, MK group, OM-85BV group, and MK+OM-85BV group, with 8 guinea pigs in each group. Intraperitoneal injection of 10% ovalbumin (OVA) for sensitization and aerosol inhalation of 1% OVA for challenge were performed to establish a model of airway remodeling of asthma in all of the groups apart from the normal control group, which were treated with normal saline. In the stage of challenge by aerosol inhalation, the guinea pigs in the MK, OM-85BV, and MK+OM-85BV groups were given corresponding suspension by gavage, and those in the normal control and asthma groups were given an equal volume of normal saline by gavage. Bronchoalveolar lavage fluid (BALF) of the guinea pigs was collected within 24 hours after challenge, and ELISA was used to measure the levels of TGF-ß1 and Smad7 in BALF. The guinea pigs were sacrificed and the pathological section of lung tissue was prepared to observe the degree of airway remodeling. An image analysis technique was used to measure perimeter of the basement membrane (Pbm), total bronchial wall area (Wat), and airway bronchial smooth muscle area (Wam). Pearson linear regression was used to investigate the correlation between two variables. RESULTS: According to the lung pathological section, compared with the normal control group, the asthma, MK, OM-85BV, and MK+OM-85BV groups had significant thickening of bronchial smooth muscle and alveolar wall, significantly higher Wat/Pbm and Wam/Pbm, a significantly higher level of TGF-ß1, and a significantly lower level of Smad7 (P<0.05). Compared with the asthma group, the MK, OM-85BV, and MK+OM-85BV groups had a significant improvement in pathological injury, significantly lower Wat/Pbm and Wam/Pbm, a significantly lower level of TGF-ß1, and a significantly higher level of Smad7 (P<0.05). The MK+OM-85BV group had significantly greater improvements than the MK group and the OM-85BV group (P<0.05). The expression of TGF-ß1 was negatively correlated with that of Smad7 and positively correlated with Wat/Pbm and Wam/Pbm, and the expression of Smad7 was negatively correlated with Wat/Pbm and Wam/Pbm (P<0.01). CONCLUSIONS: MK and OM-85BV, used alone or in combination, can reduce airway remodeling in guinea pigs with asthma, and MK combined with OM-85BV has the best effect, possibly by reducing TGF-ß1 expression, increasing Smad7 expression, and improving the TGF-ß1/Smad7 imbalance.
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Remodelación de las Vías Aéreas (Respiratorias) , Asma , Acetatos , Animales , Extractos Celulares , Ciclopropanos , Cobayas , Pulmón , Masculino , Ovalbúmina , Quinolinas , Sulfuros , Factor de Crecimiento Transformador beta1RESUMEN
The re-emerging human adenovirus types HAdV7, HAdV14, and HAdV55 of species B have caused severe lower respiratory tract diseases and even deaths during recent outbreaks. However, no adenovirus vaccine or therapeutic has been approved for general use. These adenoviruses attach to host cells via the knob domain of the fiber, using human desmoglein 2 as the primary cellular receptor. In this study, a recombinant HAdV11 fiber knob trimer (HAdV11FK) expressed in E. coli was shown to induce broadly neutralizing antibodies against HAdV11, -7, -14p1, and -55 in mice. Using HAdV11FK as an antigen, three monoclonal antibodies, 6A7, 3F11, and 3D8, with high neutralizing activity were generated. More importantly, the results of in vitro neutralization assays demonstrated that 3F11 and 3D8 cross-neutralized HAdV11, -7, and -55, but not HAdV14p1. The amino acids 251KE252 within the F-G loop may be the crucial amino acids in the conformational epitope recognized by 3F11, which is common to HAdV11, -7, -14p, and -55, but is not present in HAdV14p1 and HAdV3. A two-amino-acid deletion in the HAdV14p1 structure breaks the short alpha helix (248SREKE252) that is present in the HAdV7, -11, -55, and -14p fiber knob structures. Our findings add to the knowledge of adenovirus fiber structure and antibody responses and are important for the design of adenovirus vaccines and antiviral drugs with broad activity.
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Adenovirus Humanos/inmunología , Anticuerpos Monoclonales/inmunología , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Proteínas de la Cápside/inmunología , Anticuerpos Monoclonales/aislamiento & purificación , Anticuerpos Neutralizantes/aislamiento & purificación , Anticuerpos Antivirales/aislamiento & purificación , Enfermedades Transmisibles Emergentes/virología , Desmogleína 2/metabolismo , Epítopos/química , Epítopos/inmunología , Escherichia coli/genética , Humanos , Pruebas de Neutralización , Proteínas Recombinantes/inmunologíaRESUMEN
Human adenoviruses types 3 (HAdV-3), 7 (HAdV-7) and 55 (HAdV-55) are major pathogens of acute respiratory infections (ARI) in children and adults. More than one type of HAdV can infect patients simultaneously, and the infections are sometimes fatal. However, there is currently no vaccine approved for general use in children and adults. Thus, development of a multivalent HAdV vaccine to combat HAdV infection becomes imperative. In this study, we constructed a new recombinant trivalent human adenovirus vaccine (rAdMHE3-h55), which expresses the hexon protein of HAdV-55 in the E3 region of rAdMHE3, a previously prepared bivalent vaccine candidate against HAdV-3 and HAdV-7. The results of in vitro neutralization assays indicate that rAdMHE3-h55 can induce the production of neutralizing antibodies against HAdV-3, HAdV-7, and HAdV-55 in mice. Furthermore, immunization with the recombinant trivalent vaccine candidate completely protected the mice challenged with HAdV-3, HAdV-7, orHAdV-55, respectively, showing lower lung viral loads and less lung Pathological changes was compared with those in unvaccinated mice. The current findings contribute to the development of a new adenovirus vaccine candidate and also advance this construction method for the generation of recombinant adenovirus vaccines. In conclusion, our recombinant trivalent vaccine rAdMHE3-h55 can provides protection against challenge with HAdV-3, HAdV-7, or HAdV-55 in mice. Future work of optimizing this vaccine candidate may lead to a more effective way of preventing respiratory diseases caused by common human adenoviruses.
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Infecciones por Adenovirus Humanos/prevención & control , Vacunas contra el Adenovirus/inmunología , Adenovirus Humanos/inmunología , Vacunas Sintéticas/inmunología , Infecciones por Adenovirus Humanos/inmunología , Infecciones por Adenovirus Humanos/patología , Adenovirus Humanos/clasificación , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Modelos Animales de Enfermedad , Femenino , Orden Génico , Vectores Genéticos/genética , Vectores Genéticos/inmunología , Humanos , Inmunidad Activa , Ratones , Infecciones del Sistema Respiratorio/prevención & control , Infecciones del Sistema Respiratorio/virología , VacunaciónRESUMEN
OBJECTIVE: To study the influence of glycyrrhetinic acid (GA) on bronchial asthma (BA) smooth muscle proliferation and apoptosis as well as inflammatory factor expression and its molecular mechanism. METHODS: Male SD guinea pigs were selected and made into asthma models, bronchial asthma smooth muscle cells were cultured and divided into BA group, GA group and GA + LM group that were treated with serum-free RPMI1640 culture medium, serum-free RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid, serum-free RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid and 100 ng/mL LM22B-10 respectively; normal guinea pigs were collected and bronchial smooth muscle cells were cultured as control group. The cell proliferation activity as well as the expression of proliferation and apoptosis genes, inflammatory factors and p-ERK1/2 was determined. RESULTS: Proliferation activity value and mRNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in BA group were significantly higher than those of control group while mRNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly lower than that of control group (P < 0.05); proliferation activity value and mRNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in GA group were significantly lower than those of BA group (P < 0.05) while the mRNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly higher than those of BA group (P < 0.05); proliferation activity value and mRNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40 of airway smooth muscle cell in GA + LM group were significantly higher than those of GA group (P < 0.05) while mRNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly lower that of GA group (P < 0.05). CONCLUSION: GA can inhibit the proliferation of bronchial smooth muscle cells and reduce the expression of inflammatory factors by inhibiting the phosphorylation of ERK1/2.
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OBJECTIVE: To investigate the distribution characteristics of Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Legionella pneumophila (LP) in hospitalized children with acute respiratory tract infection (ARTI). METHODS: A total of 13 198 hospitalized children with ARTI were enrolled as study subjects. Whole blood and urine were collected. The passive agglutination was used to detect serum MP-IgM, ELISA was used to detect serum CP-IgM, and immunochromatography was performed to detect urinary LP antigen. RESULTS: Among the 13 198 hospitalized ARTI children, the detection rates of MP, CP, and LP were 25.31%, 12.74% and 3.27%, suggesting that MP had the highest detection rate (P<0.0125). The detection rates of MP in 2013 and 2014 were significantly higher than that in 2012 (P<0.0125). CP had the highest detection rate in 2013, and LP had the highest detection rate in 2014 (P<0.0125). These three pathogens were detected all around the year, and MP had the highest detection rate in all seasons (P<0.0125). The detection rate of mixed infection with three pathogens was 4.35%, and mixed infection with MP and CP was the most common (P<0.0071). Among the children in different age groups, the patients aged 5-16 years showed the highest overall detection rate of three pathogens (P<0.0071). Among the children with different types of ARTI, the children with bronchopneumonia showed the highest overall detection rate of three pathogens (P<0.0045). CONCLUSIONS: MP, CP, and LP, particularly MP, are important pathogens for children with ARTI in the local area. LP infection tends to increase year by year and should be taken seriously in clinical practice.
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Chlamydophila pneumoniae/aislamiento & purificación , Legionella pneumophila/aislamiento & purificación , Mycoplasma pneumoniae/aislamiento & purificación , Infecciones del Sistema Respiratorio/microbiología , Enfermedad Aguda , Adolescente , Niño , Niño Hospitalizado , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , EmbarazoRESUMEN
Our previous studies suggest that antibodies against ENO1 (anti-ENO1 Ab) have a protective role in patients with non-small cell lung carcinoma. In this study, we evaluated the prognostic value of anti-ENO1 Ab levels in non-small cell lung carcinoma patients undergoing surgery. Circulating levels of anti-ENO1 Ab were assessed in 85 non-small cell lung carcinoma patients before and after surgery, and were correlated with clinical outcome. After surgery, patients with a higher increase of anti-ENO1 Ab had a lower hazard ratio and a better progression-free survival. Using animal models, we demonstrated that tumor cells reduce the circulating levels of anti-ENO1 Ab through physical absorption and neutralization of anti-ENO1 Ab with surface-expressed and secreted ENO1, respectively. Mice transplanted with ENO1-overexpressing tumors generated ENO1-specific regulatory T cells to suppress the production of anti-ENO1 Ab. Our results suggest that the increase of anti-ENO1 Ab may reflect anti-tumor immune responses and serve as a prognostic marker in postoperative lung cancer patients.
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Anticuerpos Antineoplásicos/sangre , Biomarcadores de Tumor/inmunología , Carcinoma de Pulmón de Células no Pequeñas/inmunología , Proteínas de Unión al ADN/inmunología , Neoplasias Pulmonares/inmunología , Fosfopiruvato Hidratasa/inmunología , Proteínas Supresoras de Tumor/inmunología , Anciano , Animales , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/sangre , Carcinoma de Pulmón de Células no Pequeñas/sangre , Carcinoma de Pulmón de Células no Pequeñas/mortalidad , Supervivencia sin Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Estimación de Kaplan-Meier , Neoplasias Pulmonares/sangre , Neoplasias Pulmonares/mortalidad , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Persona de Mediana Edad , PronósticoRESUMEN
PURPOSE: The present study investigated whether a high-protein diet affects spatial learning and memory in premature rats via modulation of mammalian target of rapamycin (mTOR) signaling. METHODS: Pre- and full-term Sprague-Dawley pups were fed a normal (18% protein) or high-protein (30% protein) diet (HPD) for 6 or 8 weeks after weaning. Spatial learning and memory were tested in the Morris water maze at week 6 and 8. The activation of mTOR signaling pathway components was evaluated by western blotting. RESULTS: Spatial memory performance of premature rats consuming a normal and HPD was lower than that of full-term rats on the same diet at 6 weeks, and was associated with lower levels of ribosomal protein S6 kinase p70 subtype (p70S6K) and initiation factor 4E-binding protein 1 (4EBP1) phosphorylation in the hippocampus. Spatial memory was improved in 8-week-old premature rats on an HPD as compared to those on a normal diet. Premature rats on an HPD had p70S6K and 4EBP1 phosphorylation levels in the hippocampus that were comparable to those of full-term rats on an HPD. CONCLUSION: Long-term consumption of a protein-rich diet can restore the impairment in learning and memory in pre-term rats via upregulation of mTOR/p70S6K signaling.
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Proteínas en la Dieta/administración & dosificación , Transducción de Señal , Aprendizaje Espacial/fisiología , Memoria Espacial/fisiología , Animales , Animales Recién Nacidos , Hipocampo/metabolismo , Aprendizaje por Laberinto/fisiología , Ratas , Ratas Sprague-Dawley , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
OBJECTIVE: To establish a scientific method for identitication and evaluation of the Tibetan prescription Jia Ga Song Tang. METHODS: Volatile oil was extracted by water steam distillation and analyzed by GC-MS. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied to the samples for chemical fingerprint pattern recognition research. RESULTS: 16 samples according to hierarchical cluster analysis (HCA) and principal component analysis (PCA) were divided into two classes, and results from two recognition analysis methods had good consistency. CONCLUSION: GC-MS-pattern recognition method was a kind of scientific, accurate and effective method for the quality evaluation of Jia Ga Song Tang.
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Medicamentos Herbarios Chinos/química , Cromatografía de Gases y Espectrometría de Masas , Aceites Volátiles/química , Control de Calidad , Análisis por Conglomerados , Medicamentos Herbarios Chinos/normas , Aceites Volátiles/normas , Análisis de Componente Principal , VaporRESUMEN
Diammonium glycyrrhizinate (DG), an active compound extracted and purified from liquorices root, has been reported to exhibit antioxidant and anti-inflammatory properties. The aim of this study was to investigate the effect and underlying mechanisms of DG on the hepatotoxicity induced by valproic acid (VPA). DG at the dose of 60mg/kg was orally administered with VPA (100mg/kg) to mice once daily for 14 consecutive days. DG treatment attenuated VPA-induced liver dysfunction, structural damage, glutathione depletion and decrease in antioxidant enzymes in BALB/C mice. DG prevented VPA-induced depletion of cytosolic nuclear factor E2-related factor 2 (Nrf2) and suppression of nuclear translocation of Nrf2, which, in turn, up-regulated phase II/antioxidant enzyme activities. The effects of VPA and DG on Nrf2 expression in HepG2 cells were in consistent with that of mice. Furthermore, an increase in the nuclear levels of nuclear factor-kappaB (NF-κB) was observed in the livers of VPA-treated mice that coincided with induction of inflammatory cytokines. In contrast, DG inhibited NF-κB translocation and that subsequently decreased inflammatory cytokines. Taken together, these results demonstrate that DG attenuates VPA-induced liver injury through increasing the expression of Nrf2 mediated phase II/antioxidant enzymes and simultaneously decreasing the expression of inflammatory mediators.