RESUMEN
Vitamin D (25(OH)D3) is an essential nutrient that plays a role in the immune system. Serum 25(OH)D3 is found to be associated with asthma. However, the role of vitamin D in obese asthma remains unclear. Therefore, we investigated the association between vitamin D levels and asthma outcomes in a murine model of obese asthma. We also evaluated NLRP3 inflammasome activity in the pathogenesis of obese asthma. We divided 20 male Balb/c mice (3-4 weeks old) into 4 groups: normal control, asthma, obese, and obese asthma and developed an obese asthma mouse model. Airway hyperreactivity, cytokine concentrations, 25(OH)D3 levels, NLRP3 mRNA and IL-1β mRNA expressions were measured. Lung histology and bronchoalveolar lavage fluid (BALF) cell count were also determined. Obese asthma mice showed a significant increase in airway hyper-responsiveness, airway inflammation, pro-inflammatory cytokine levels and NLRP3 mRNA, IL-1β mRNA expression. Both asthma and obese groups had lower 25(OH)D3 levels. Vitamin D levels in obese asthma were the lowest among all groups. Vitamin D levels correlated negatively with body weight, lung resistance levels at 25 mg/mL of methacholine, total inflammatory cells, and IL-1β and IL-17 concentrations in BALF. These data demonstrated an association between serum vitamin D levels and outcomes of obese asthma, and indicated that NLRP3 inflammasome may play a role in this disorder.
Asunto(s)
Animales , Masculino , Asma/fisiopatología , Asma/metabolismo , Colecalciferol/sangre , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Obesidad/fisiopatología , Obesidad/metabolismo , Asma/patología , Factores de Tiempo , Peso Corporal , Ensayo de Inmunoadsorción Enzimática , Líquido del Lavado Bronquioalveolar , Citocinas/análisis , Citocinas/metabolismo , Modelos Animales de EnfermedadRESUMEN
Vitamin D (25(OH)D3) is an essential nutrient that plays a role in the immune system. Serum 25(OH)D3 is found to be associated with asthma. However, the role of vitamin D in obese asthma remains unclear. Therefore, we investigated the association between vitamin D levels and asthma outcomes in a murine model of obese asthma. We also evaluated NLRP3 inflammasome activity in the pathogenesis of obese asthma. We divided 20 male Balb/c mice (3-4 weeks old) into 4 groups: normal control, asthma, obese, and obese asthma and developed an obese asthma mouse model. Airway hyperreactivity, cytokine concentrations, 25(OH)D3 levels, NLRP3 mRNA and IL-1ß mRNA expressions were measured. Lung histology and bronchoalveolar lavage fluid (BALF) cell count were also determined. Obese asthma mice showed a significant increase in airway hyper-responsiveness, airway inflammation, pro-inflammatory cytokine levels and NLRP3 mRNA, IL-1ß mRNA expression. Both asthma and obese groups had lower 25(OH)D3 levels. Vitamin D levels in obese asthma were the lowest among all groups. Vitamin D levels correlated negatively with body weight, lung resistance levels at 25 mg/mL of methacholine, total inflammatory cells, and IL-1ß and IL-17 concentrations in BALF. These data demonstrated an association between serum vitamin D levels and outcomes of obese asthma, and indicated that NLRP3 inflammasome may play a role in this disorder.
Asunto(s)
Asma/metabolismo , Asma/fisiopatología , Colecalciferol/sangre , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Obesidad/metabolismo , Obesidad/fisiopatología , Animales , Asma/patología , Peso Corporal , Líquido del Lavado Bronquioalveolar , Citocinas/análisis , Citocinas/metabolismo , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Recuento de Leucocitos , Pulmón/patología , Masculino , Ratones Endogámicos BALB C , Obesidad/patología , Reacción en Cadena en Tiempo Real de la Polimerasa , Factores de TiempoRESUMEN
Like other developing countries, China was reported to have a relatively high seroprevalence of anti-hepatitis A antibodies (anti-HAV). However, no studies have evaluated the prevalence of anti-HAV and HAV RNA among voluntary blood donors with or without elevated serum alanine transaminase (ALT) levels. Anti-HAV antibodies were detected using an enzyme-linked immunosorbent assay, and reverse transcription quantitative polymerase chain reaction was carried out for detection of HAV RNA. In the current study, we analyzed a total of 450 serum samples with elevated ALT levels (≥40 U/L) and 278 serum samples with non-elevated ALT levels. Seroprevalence rates of anti-HAV were 51.6% in donors with elevated ALT and 41.4% in donors with non-elevated ALT; however, none of the samples was positive for HAV RNA. The results of our study showed lower seroprevalence rates of anti-HAV in blood donors (irrespective of ALT levels) than those in published data on Chinese populations. Although donors with elevated ALT had statistically higher prevalence rates of anti- HAV than did those with non-elevated ALT, none of the serum samples had detectable levels of the active virus. In conclusion, our results demonstrate that the transmission of hepatitis A by blood transfusion will occur rarely.
Asunto(s)
Virus de la Hepatitis A Humana/inmunología , Hepatitis A/epidemiología , Hepatitis A/inmunología , Adolescente , Adulto , China/epidemiología , Femenino , Hepatitis A/virología , Anticuerpos de Hepatitis A/sangre , Humanos , Pruebas de Función Hepática , Masculino , Persona de Mediana Edad , Estudios Seroepidemiológicos , Carga Viral , Adulto JovenRESUMEN
Hemophilia A (HA) is an inherited X-linked bleeding disorder caused by mutations in the factor VIII gene. Prenatal detection in female carriers from families with HA is important to reduce the number of HA patients. The purpose of this study was to detect carriers in families with HA from Sichuan, China, using linkage analysis and a direct genotyping method. A total of 18 HA families were studied. Using a combination of intron 22 inversion, intron 1 inversion, the BclI polymorphic site in intron 18, the HindIII polymorphic site in intron 19, and dinucleotide CA-repeat markers in introns 1, 13, 22, and 24, we were able to detect HA in 88.9% (16/18) of the families studied. HA was detected in the remaining two families by direct genotyping. This study gave the participants a good understanding of their genetic condition and gave us a preliminary understanding of the prevalence of each mutation in Sichuan HA patients.
Asunto(s)
Factor VIII/genética , Tamización de Portadores Genéticos , Ligamiento Genético , Hemofilia A/genética , Adulto , China , Inversión Cromosómica/genética , Femenino , Genotipo , Heterocigoto , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Intrones/genética , Masculino , MutaciónRESUMEN
The purpose of this study was to examine the changes of cellular cholesterol efflux from macrophages in patients with type II diabetes mellitus (DM), and to determine the expression of CYP7A1, ABCG5, and LXRß therein. We recruited 30 patients with type II DM (including 15 patients complicated with coronary heart disease and 15 patients with DM only) and 15 normal controls for this study. Peripheral blood monocytes were isolated for macrophage culture. The mRNA and protein expression levels of CYP7A1, ABCG5, and LXRß were determined using real-time polymerase chain reaction and western blot. The macrophage cholesterol efflux rate was determined with 10% autoserum and standard serum as receptors. We determined that the expression levels of macrophage CYP7A1 mRNA and protein in the type II DM group were significantly lower than those in the control group, but no differences were found in the ABCG5 and LXRß expression levels between the groups. The macrophage cholesterol efflux rate in the patients with type II DM was also significantly decreased compared with that of the normal control subjects (P < 0.01). Furthermore, CYP7A1 mRNA expression and macrophage cholesterol efflux rate were significantly positively correlated. In summary, this study demonstrated that the macrophage cholesterol efflux in patients with type II DM was significantly reduced, and that this reduction was associated with the down-regulation of CYP7A1 expression.
Asunto(s)
Colesterol 7-alfa-Hidroxilasa/genética , Colesterol/metabolismo , Enfermedad Coronaria/enzimología , Diabetes Mellitus Tipo 2/enzimología , Macrófagos/enzimología , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 5 , Transportadoras de Casetes de Unión a ATP/genética , Transportadoras de Casetes de Unión a ATP/metabolismo , Adulto , Estudios de Casos y Controles , Células Cultivadas , Colesterol/sangre , Colesterol 7-alfa-Hidroxilasa/metabolismo , Enfermedad Coronaria/sangre , Enfermedad Coronaria/etiología , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/complicaciones , Regulación hacia Abajo , Represión Enzimática , Femenino , Humanos , Lipoproteínas/genética , Lipoproteínas/metabolismo , Receptores X del Hígado , Macrófagos/metabolismo , Masculino , Persona de Mediana Edad , Receptores Nucleares Huérfanos/genética , Receptores Nucleares Huérfanos/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Cohn fraction IV (CFIV) is a byproduct of a plasma fractionation process known as the Cohn process. It is an inexpensive source of protein C, retaining about 90% of protein C (PC) in human plasma. We investigated whether PC is affected during the Cohn process and evaluated correlations among coagulant activity, amidolytic activity and PC antigen during the Cohn process. CFIV was redissolved with citrate-buffered saline for 5 h at 4°C, and then centrifuged at 3500 g for 40 min at 4°C. Functional anticoagulant activity was measured with a one-stage coagulation method based on activated partial thromboplastin time. The functional amidolytic activity of PC was determined using chromogenic substrate assay, and measurement of PC antigen was performed by ELISA. In CFIV, anticoagulant activity declined significantly, with a loss of >80%, while amidolytic activity was not significantly altered, compared to PC antigen. Prior to the Cohn process, high-rank correlations were observed in cryosupernatant, with rs = 0.921 for anticoagulant and amidolytic activities (P = 0.009), 0.896 for anticoagulant activity and antigen (P = 0.014) and 0.832 for amidolytic activity and antigen (P = 0.031). After the Cohn process in CFIV, there was also a high correlation between amidolytic activity and antigen (rs = 0.782, P = 0.038). There were no significant correlations between anticoagulant activity and antigen (rs = 0.223, P = 0.653), or anticoagulant and amidolytic activity (rs = 0.236, P = 0.675). We conclude that the Cohn process significantly influences the anticoagulant activity of PC. Compared to the antigen, PC lost greater than 80% of its anticoagulant activity, but retained its amidolytic activity, during the Cohn process.
Asunto(s)
Anticoagulantes/sangre , Coagulación Sanguínea/genética , Proteínas Sanguíneas/metabolismo , Proteína C/metabolismo , Antígenos/sangre , Proteínas Sanguíneas/genética , Ensayo de Inmunoadsorción Enzimática , Humanos , Proteína C/genéticaRESUMEN
This study aimed to investigate the value of magnetic resonance spectroscopy (MRS) imaging in assessing nasopharyngeal carcinoma radiotherapy during the early delayed reaction period. Eighty cases of nasopharyngeal cancer treated with radiotherapy within the same period underwent MRS imaging before or after radiotherapy. Of the 80 cases, 47 underwent MRS imaging on the 3rd, 4th, 6th, and 12th months after radiotherapy. The trends of the primary metabolite concentration at different time points were monitored and compared with the corresponding data after radiotherapy. Repeated measures analysis of variance was performed. At the end of radiotherapy, the N-acetyl aspartate (NAA)/creatine (Cr), choline (Cho)/Cr, and NAA/Cho ratios were reduced to the lowest levels after 3 months. However, increasing trends were observed from the 4th to the 12th month. On the 12th month, stable levels were reached with statistically significant differences (F = 316.02, 53.84, 286.68; P < 0.01). MRS reflected the radiation injury-repair process in the brain of a nasopharyngeal cancer patient during early delayed reaction. This non-invasive monitoring of changes in brain tissue metabolite concentrations provides valuable information for prognosis.
Asunto(s)
Lesiones Encefálicas/diagnóstico , Imagen por Resonancia Magnética/métodos , Espectroscopía de Resonancia Magnética/métodos , Neoplasias Nasofaríngeas/radioterapia , Traumatismos por Radiación/diagnóstico , Adulto , Ácido Aspártico/análogos & derivados , Ácido Aspártico/metabolismo , Lesiones Encefálicas/etiología , Lesiones Encefálicas/metabolismo , Colina/metabolismo , Creatina/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Traumatismos por Radiación/etiología , Traumatismos por Radiación/metabolismo , Radioterapia/efectos adversos , Radioterapia/métodos , Factores de TiempoRESUMEN
Residual activated coagulation factor XI (FXIa) has been suggested to play an important role in thromboembolic events associated with the use of intravenous immunoglobulin (IVIG) lots. This study investigated the predominant plasma proteases in 42 IVIG lots from 4 Chinese manufacturers. In one-stage clotting assays, the procoagulant activities of factors II, VII, IX, X, XI, and XII were quantified. Non-activated partial thromboplastin time and a modified thrombin generation test served as global and FXIa-specific clotting assays, respectively. We found that coagulation factor clotting activities of the 42 IVIG lots were below the detection limit of the assays, except for the products of manufacturer B (lots of 2010), in which 0.030 to 0.032 IU/mL FXI:C were detected. The peak time of thrombin using a thrombin generation test was greater than 35 min, the relevant amount of FXIa was below 0.37 nM, and non-activated partial thromboplastin time was greater than 200 s. Consequently, the 42 IVIG lots showed non-significant procoagulant potential. Further study is required to determine whether a program for FXIa determination in IVIG products should be launched in China.
Asunto(s)
Contaminación de Medicamentos , Factor XI/análisis , Inmunoglobulinas Intravenosas/química , China , Industria Farmacéutica , HumanosRESUMEN
Human coagulation factor VII (FVII) plays an important role in the blood coagulation process and exists in micro amounts in human plasma; therefore, any attempt at the large-scale production of FVII in significant quantities is challenging. The purpose of this study was to express and obtain biologically active recombinant FVII (rFVII) from Chinese hamster ovary K1 (CHO-K1) cells. The full-length FVII cDNA was isolated from a HepG2 cell line and then subcloned in pcDNA3.1 to construct an expression vector, pcDNA-FVII. CHO-K1 cells were transfected with 1 µg pcDNA-FVII. The cell line that stably expressed secretory FVII was screened using 900 µg/mL G418. The FVII copy number in CHO-K1 cells was detected by quantitative polymerase chain reaction (qPCR). The rFVII was purified in ligand affinity chromatography medium. The purified protein was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The biological activity of the purified FVII protein was determined by a prothrombin time assay. Three cell lines that permanently expressed rFVII were screened. The qPCR results demonstrated that each CHO-K1 cell harbored two FVII DNA copies. The SDS-PAGE and Western blot analysis showed that the purified protein was about 50 kDa. The purity of the target protein was 95%. The prothrombin time assay indicated that the FVII-specific activity of rFVII was 2573 ± 75 IU/mg. This method enabled the fast preparation of high-purity rFVII from CHO-K1 cells, and the purified protein had good biological activity.