RESUMEN
OBJECTIVE: To study the molecular genetic mechanism and genetic diagnosis of pyruvate dehydrogenase complex deficiency (PHD), and to provide a basis for genetic counseling and prenatal genetic diagnosis of PHD. METHODS: Polymerase chain reaction (PCR) was performed to amplify the 11 exons and exon junction of the PDHA1 gene from a child who was diagnosed with PHD based on clinical characteristics and laboratory examination results. The PCR products were sequenced to determine the mutation. An analysis of amino acid conservation and prediction of protein secondary and tertiary structure were performed using bioinformatic approaches to identify the pathogenicity of the novel mutation. RESULTS: One novel duplication mutation, c.1111_1158dup48bp, was found in the exon 11 of the PDHA1 gene of the patient. No c.1111_1158dup48bp mutation was detected in the sequencing results from 50 normal controls. The results of protein secondary and tertiary structure prediction showed that the novel mutation c.1111 _1158dup48bp led to the duplication of 16 amino acids residues, serine371 to phenylalanine386, which induced a substantial change in protein secondary and tertiary structure. The conformational change was not detected in the normal controls. CONCLUSIONS: The novel duplication mutation c.1111_1158dup48bp in the PDHA1 gene is not due to gene polymorphisms but a possible novel pathogenic mutation for PHD.
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Mutación , Piruvato Deshidrogenasa (Lipoamida)/genética , Enfermedad por Deficiencia del Complejo Piruvato Deshidrogenasa/genética , Secuencia de Aminoácidos , Humanos , Lactante , Masculino , Datos de Secuencia Molecular , Conformación Proteica , Piruvato Deshidrogenasa (Lipoamida)/químicaRESUMEN
AIM: To observe the anti-oxidative activity and adverse laxative effect of raw, traditional processed and fermented products of Polygoni Multiflori Radix (PMR), and furthermore, to evaluate the fermentation method used in the processing procedure of PMR. METHODS: In vitro ferric reducing antioxidant power (FRAP) assay was carried out to evaluate the anti-oxidative activity. Modulation of normal defecation and effect on gastrointestinal motility in mice were carried out to investigate their adverse laxative effect. RESULTS: Fermented PMR induced less severe laxative adverse effect than Polygoni Multiflori Radix Praeparata (PMRP). PMR fermented with Rhizopus sp. (FB) could modulate the defecation significantly. The gastrointestinal motility was inhibited by PMRP and PMR fermented with Rhizopus oryzae (FA). FA and FB showed better antioxidant activity than PMRP in 50% and 95% ethanol group. Contents of 2, 3, 5, 4'-tetrahydroxy-stilbene-2-O-ß-D-glucoside (TSG) were reduced significantly after traditional processing but maintained after fermentation. Emodin and physcion were increased after traditional processing and fermented with Rhizopus oryzae. CONCLUSION: All processing procedure, including fermentation, might reduce its anti-oxidative activity. However, most of the processed products could lessen the adverse effect on gastrointestinal tract compared to PMR. Fermentation with Rhizopus oryzae was considered as a promising processing method of PMR.
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Antioxidantes/farmacología , Defecación/efectos de los fármacos , Motilidad Gastrointestinal/efectos de los fármacos , Tracto Gastrointestinal/efectos de los fármacos , Laxativos/efectos adversos , Extractos Vegetales/farmacología , Polygonum/química , Animales , Emodina/efectos adversos , Emodina/análogos & derivados , Emodina/farmacología , Femenino , Fermentación , Masculino , Ratones , Ratones Endogámicos , Extractos Vegetales/efectos adversos , Raíces de Plantas/química , Polygonum/efectos adversos , RhizopusRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Radix Polygoni Multiflori (RPM) and Radix Polygoni Multiflori Praeparata (RPMP) were traditionally widely used as Chinese herbal medicine. However, liver adverse reactions caused by RPM or RPMP were frequently reported all around the world recent years. The aim of this study was to study the cytotoxicities of RPM, RPMP and their major constituents on human liver cell L-02 simultaneously. MATERIALS AND METHODS: Multi-assays, including MTT assay, neutral red uptake (NRU) assay, LDH leakage percentage and liver enzyme secretion (AST, ALT and ALP) were used. Cytotoxicities of major chemical constituents of RPM, 2, 3, 5, 4'-tetrahydroxy-stilbene-2-O-ß-D-glucoside (TSG), physcion and emodin, were tested. The cytotoxicities of water, 50% ethanol and 95% ethanol extractions of RPM and RPMP were tested. HPLC-DAD analysis was carried to reveal the content change of TSG, physcion and emodin after the processing procedure. RESULTS: The TD(50) of TSG, physcion and emodin in MTT assay were >10,000 µM, 2853.61 µM and 520.37 µM. In the NRU assay, the TD(50) of TSG, physcion and emodin were much smaller (1401.53 µM, 1140.00 µM, and 3.80 µM). Emodin induced much severe liver enzyme secretion than TSG and physcion. Cell proliferation and LDH leakage rate showed no difference between RPM and RPMP extractions, but ALP, AST and ALT secretions in RPMP extractions were significant lower than that of PMR groups. Water extractions of RPM and RPMP were less toxic than any other solvent in most of the assays. Positive correlation was found between the TSG/emodin ratio and MTT survival rate. The emodin/physcion ratio also showed positive correlation with the LDH leakage percentage. CONCLUSIONS: In conclusion, Radix Polygonum multiflorum and Radix Polygonum multiflorum Praeparata were not liver injure inducing in our in vitro assays. However, the processing produce of RPM could reduce its effect on both cell proliferation and enzyme secretion of liver cell. Judging from cell proliferation, integrity of cell membrane and enzyme secretion, three major chemical constituents of RPM: TSG, physcion and emodin showed no, moderate and severe cytotoxicity against human liver cell line L-02 respectively. Chemical constituents-cytotoxicity relationship investigation revealed that TSG and physcion probably had attenuating effect to emodin. The attenuating mechanisms were still under investigation.
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Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Medicamentos Herbarios Chinos/toxicidad , Emodina/análogos & derivados , Glucósidos/toxicidad , Hígado/efectos de los fármacos , Polygonaceae , Estilbenos/toxicidad , Alanina Transaminasa/metabolismo , Fosfatasa Alcalina/metabolismo , Aspartato Aminotransferasas/metabolismo , Línea Celular , Proliferación Celular/efectos de los fármacos , Enfermedad Hepática Inducida por Sustancias y Drogas/enzimología , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/aislamiento & purificación , Emodina/aislamiento & purificación , Emodina/toxicidad , Glucósidos/aislamiento & purificación , Humanos , L-Lactato Deshidrogenasa/metabolismo , Hígado/enzimología , Hígado/patología , Raíces de Plantas , Plantas Medicinales , Polygonaceae/química , Solventes/química , Estilbenos/aislamiento & purificaciónRESUMEN
OBJECTIVE: To analyze the clinical and genetic characteristics of three children with ornithine carbamoyltransferase deficiency(OTCD), and to provide a practical method for gene diagnosis and genetic counseling of the disease. METHODS: All exons of the ornithine carbamoyltransferase (OTC) gene were screened by polymerase chain reaction-DNA direct sequencing in the three OTCD patients. RESULTS: One patient firstly presented as vomiting at 6 month of age. A missense mutation of T262I was detected. His mother had the same mutation without any clinical symptoms. The second patient presented as restlessness, and had a missense mutation of R277W. Gene analysis of his parents was not available. The third patient presented as neonatal lethargy, harbored a missense mutation of I172M. His mother had the same mutation without any clinical symptoms. CONCLUSION: Gene mutation analysis is a feasible way for diagnosing OTCD. Patients with I172M mutation present symptom early, while those with T262I and R277W mutations manifest symptoms later. Gene mutation analysis will be important for asymptomatic and prenatal diagnosis and genetic counseling.
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Mutación/genética , Enfermedad por Deficiencia de Ornitina Carbamoiltransferasa/genética , Ornitina Carbamoiltransferasa/genética , Secuencia de Bases , Niño , Exones , Humanos , Lactante , Recién Nacido , Masculino , Enfermedad por Deficiencia de Ornitina Carbamoiltransferasa/patologíaRESUMEN
OBJECTIVE: To understand the characteristics of adenovirus infection in hospitalized children with pneumonia in Guangzhou area. METHODS: The infection rate, hospitalization time and hospitalization expenses of adenovirus-infected hospitalized children with pneumonia in Guangzhou area from 2005 to 2007 were analyzed. RESULTS: The total adenovirus infection rate was 6.04% in these children, with a male to female ratio of 1.47:1, showing significantly higher infection rate in female (7.92%) than in male patients (5.21%, P<0.05). The hospital stay and hospitalization costs between male and female children showed no significant difference (P>0.05). Adenovirus-infected children from birth to six years old accounted for 90.50% of the total adenovirus-infected children, and the infection rate in 0 to 1-year-old children (3.71%) was significantly lower than that in elder children (P<0.05). Although the infection rate in winter (8.44%) was significantly higher than that in the other seasons (P<0.05), the cases from March to August accounted for 60.11% of the total infected cases. Furthermore, the infection rate in 2007 (4.31%) was significantly lower than that in 2005 and 2006 (7.11% and 6.71%, respectively, P<0.05). CONCLUSION: Adenovirus infection is an important pathogen in hospitalized children with pneumonia in Guangzhou area, and the infection rates differed between gender, age, season and the years.
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Infecciones por Adenovirus Humanos/epidemiología , Neumonía/virología , Infecciones por Adenovirus Humanos/economía , Adolescente , Niño , Preescolar , China/epidemiología , Costo de Enfermedad , Femenino , Hospitalización , Humanos , Incidencia , Lactante , Masculino , Neumonía/economía , Estaciones del Año , Factores SexualesRESUMEN
OBJECTIVE: To study the rearrangement of immunoglobulin (Ig) heavy chain variable region (V(H)) genes in human neonates with different gestational ages (GA). METHODS: Peripheral blood from the neonates with GA of 27 weeks (4 cases), 28-32 weeks (9 cases), 33-36 weeks (12 cases), and 37-42 weeks (13 cases) was collected. RT-PCR was used to amplify the Ig V(H) gene, and the PCR products were separated by electrophoresis and analyzed using 6% denaturing PAGE gel. RESULTS: All Ig V(H) family genes had several rearranged genes in each GA group, and the neonates with different GA showed no significant difference in the median molecular weight for each rearranged Ig V(H) family gene. CONCLUSION: The neonates with GA of 27-42 weeks exhibit diversity in Ig V(H) gene rearrangement, and for the same Ig V(H) family, the median length of the arranged Ig V(H) genes is independent of the gestational age.