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Objective To investigate the effect of CYP2D6 gene polymorphism on risperidone (RISP) metabolism in schizophrenic patients. Methods CYP2D6 allele polymorphisms including*10,*4,*41 and *2 was detected by real-time fluorescent PCR in 120 schizophrenic patients who have taken risperidone continually. Alleles without SNP mutations were classified as wild-type (WT). At the same time, serum risperidone and 9-hydroxyrisperidone concentration of all patients were detected by mass spectrometric analysis. Some samples were selected for DNA sequencing of CYP2D6*10, which is the most common CYP2D6 allele in Oriental population. The 120 patients were divided into three groups according to their allele variants. Group 1 was defined as carriers of two functional alleles, group 2 was defined as carriers of one defective allele, group 3 was defined as carriers of two defective alleles. Genotype distributions, alleles frequencies, RISP, 9-oh-RISP, RISP+9-OH-RISP, 9-oh-RISP/RISP among three groups were calculated.Results Group 1 amount to 23 cases including 13 cases of WT/WT, 7 cases of*2/*2, 3 cases of WT/*2. Group 2 amount to 51 cases, including 38 cases of WT/*10, 8 cases of *2/*10, 1 case of *2/*41, 4 cases of WT/*41. Group 3 amount to 46 cases, including 44 cases of *10/*10, 2 cases of *10/*41. The*4 allele was not detected. The allele frequency of WT, *2, *10 and *41 was 29.6%, 10.8%, 56.7% and 2.9%, respectively. The 9-hydroxyrisperidone/risperidone-ratio of three groups were 15.24±5.77, 11.06±4.56 and 2.39 ± 1.06, respectively. There was a significant difference in 9-hydroxyrisperidone/risperidone-ratio between Group 3 and the first two groups (P<0.001). Conclusions The frequency of *10 allele was the highest among the subjects. The frequency of WT and*2 allele was over 95%in the population. Individuals carrying one defective allele of CYP2D6 will decrease the rate of risperidone metabolism slightly, while individuals carrying two defective alleles of CYP2D6 may decrease the rate of risperidone metabolism significantly.
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Objective To investigate the application value of apolipoprotein E (ApoE) genotyping by DNA microarray technology and the relationship between ApoE allelic frequency and serum ApoE levels in both healthy individuals and patients with Alzheimer ′s disease (AD).Methods This research is case-control study.DNA microarray was used to detect the ApoE genotypes of AD patients (n =280) and age-matched non-demented elderly control subjects ( n =230) .The cases and controls were collected in Guangzhou Huiai Hospital during July 2014 to September 2015.The accuracy of genotype results was verified by DNA sequencing.Serum ApoE levels were measured by immunoturbidimetric assay .The ApoE genotype distribution and the relationship between ApoE allelic frequency and serum ApoE levels were analyzed.The “t” test was used to compare the ApoE levels of AD patients and controls , variance analysis was used to analyze ApoE levels in the persons with different genotype .Results DNA microarray technology genotyping results were completely consistent with the results of DNA sequencing .In AD group, the ApoE genotype distribution were 2.9%(8 /280) for ε2ε3, 1.8% (5/280) for ε2ε4, 46.8% (131/280)for ε3ε3,45.4%(127 /280) for ε3ε4 and 3.1%(9 /280) for ε4ε4.While in the control group, the ApoE genotype distribution were 0.9%(2 /230) for ε2ε2, 12.6% (29/230)for ε2ε3, 1.3%(3 /230) forε2ε4, 70.0% (161 /230) for ε3ε3 and 15.2% (35 /230) for ε3ε4.The average serum concentrations of ApoE were (33.29 ±10.87)mg/L in AD patients and (41.28 ±10.95)mg/L in the controls.Among all participants, the average serum levels of ApoE were (50.86 ±6.21) mg/L for ε2 carriers, (38.78 ± 12.07)mg/L for ε3 carriers and (30.47 ±7.68)mg/L for ε4 carriers.In AD group,ApoE level of ε2, ε3,ε4 carriers is (50.31 ±9.08)mg/L, (38.30 ±7.60) mg/L and (32.86 ±5.93)mg/L respectively.In the control group, the ApoE level of ε2, ε3, ε4 carriers is (51.00 ±5.53)mg/L, (41.01 ±10.09)mg/L and (32.86 ±5.93)mg/L respectively.The ApoE levels of persons with different ApoE alleles are ε2 >ε3 >ε4. The difference is significant (F =89.6, P 0.05) .Conclusions DNA microarray technology possesses high efficiency and favorable accuracy.The ε2 allele is associated with a higher ApoE concentration , ε3 allele with a mediate concentration and ε4 allele with a lowest concentration.Serum concentrations of ApoE showed no significant difference between AD patients and the healthy groups who have the same genotype .The primary cause of the low serum ApoE levels in AD patients is that the ApoE ε4 allelic frequencies of them are higher than that of the healthy persons.
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Objective To investigate the relationship of apolipoprotein E (ApoE) allelic frequency and serum lipid levels in patients with Alzheimer′s disease (AD). Methods DNA microarray was used to detect the ApoE genotypes of AD patients (n = 200) and age-matched non-demented elderly control subjects (n = 159). Serum lipid levels was measured by Immunoturbidimetric assay at the same time. We analyzed the ApoE genotype distribution and the relationship of apolipoprotein E ( ApoE ) allelic frequency and serum lipid levels . Results The ApoE ε4 allelic frequencies (25.5%) in AD group is higher than that of the control group (7.9%) (P 0.05). Conclusion The ApoE levels are negatively related to ApoE ε4 allele frequency and have no significant differences with the same genotype in AD and the control group,which suggests that lower serum ApoE levels in AD patients is caused by higher ApoE ε4 allelic frequency in AD than in healthy population.
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<p><b>OBJECTIVE</b>To assess the value of quantitative analysis of hepatitis B surface antigen (HBsAg) levels in the diagnosis and therapeutic evaluations in patients with chronic hepatitis B (CHB).</p><p><b>METHODS</b>According to the staging criteria defined by the American Association of Liver Diseases, 96 patients with CHB admitted in Zhujiang Hospital were classified in immune-tolerant (IT), HBeAg-positive hepatitis (EPH), inactive carrier (IC) and HBeAg-negative hepatitis (ENH) phases. Serum HBsAg, HBV-DNA and ALT levels were quantified and their correlations were evaluated in each phase of infection.</p><p><b>RESULTS</b>The mean HBsAg titers (measured in log10U/L) differed significantly between the phases of CHB (4.12 in IT, 4.02 in EPH, 2.85 in EPH, and 3.29 in ENH). The correlation coefficient of HBsAg with HBV-DNA was 0.6828 in IT, 0.5759 in EPH, 0.3280 in IC, and 0.1083 in ENH. Serum HBsAg titers were significantly higher in HBeAg-positive patients than in HBeAg-negative patients. No correlation was found between HBsAg level and ALT in each phase of CHB.</p><p><b>CONCLUSION</b>The median baseline serum HBsAg levels vary between different phases of CHB in Guangzhou, suggesting the value of HBsAg in accurate classification of hepatitis B patients and evaluation of the therapeutic effect and outcomes of the patients.</p>
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Humanos , ADN Viral , Sangre , Antígenos de Superficie de la Hepatitis B , Sangre , Virus de la Hepatitis B , Hepatitis B Crónica , Sangre , Pruebas SerológicasRESUMEN
OBJECTIVE@#To detect the expression of NGF, BDNF, NT-3 mRNA in the peripheral blood of patients with allergic rhinitis (AR). And to analyze the correlation between NGF, BDNF, NT-3 mRNA expression and the epidsode of rhinitis through Th-2 Hypothesis.@*METHOD@#This study was a group controlled trial. The expression of NGF, BDNF and NT-3 mRNA were tested by real-time quantitative RT-PCR and the concentrations of IL-4, IL-6, IL-10 and INF-alpha were tested by ELISA.@*RESULT@#The expression of NGF, BDNF and NT-3 mRNA in AR patients were 2.44, 4.46 and 1.78 times the amount of those in the healthy adults, respectively. The increased expression of NT-3 correlated positively with the scores of visual analog scale of AR. The concentrations of IL-4, IL-6 and IL-10, which were 2198 +/- 472 pg/mL, 9407 +/- 703 pg/mL and 3917 +/- 323 pg/mL respectively, were higher than those in the healthy adults. The concentration of INF-alpha was 2198 +/- 472 pg/mL and less than the healthy adults. The increased expressions of NGF, NT-3 were positively related to the increase of IL-4, IL-6 and IL-10.@*CONCLUSION@#The expressions of NGF, BDNF and NT-3 mRNA in AR patients are higher than those in the healthy adults. NGF, BDNF and NT-3 may contribute to the pathogenesis of AR. Moreover, NGF and NT-3 may induce the episode of rhinitis through Th-2 Hypothesis.
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Adolescente , Adulto , Niño , Femenino , Humanos , Masculino , Adulto Joven , Factor Neurotrófico Derivado del Encéfalo , Sangre , Estudios de Casos y Controles , Interleucina-10 , Sangre , Interleucina-4 , Sangre , Interleucina-6 , Sangre , Factor de Crecimiento Nervioso , Sangre , Factores de Crecimiento Nervioso , Sangre , Genética , Neurotrofina 3 , Sangre , ARN Mensajero , Genética , Rinitis Alérgica , Sangre , Alergia e Inmunología , Balance Th1 - Th2RESUMEN
OBJECTIVE@#To assess the expression of NGF, BDNF, NT-3 mRNA in the peripheral blood of patients with allergic rhinitis (AR). Meanwhile, to analysis whether the expression of NGF, BDNF, NT-3 mRNA correlate with the severity of rhinitis.@*METHOD@#This study is a group controlled trial, which takes the healthy adults as control group. The total RNA have been extracted from the peripheral blood of AR patients. The expression of NGF, BDNF and NT-3 mRNA have been tested by real-time quantitative RT-PCR.@*RESULT@#Comparing with the healthy adults, the expression of NGF, BDNF and NT-3 mRNA as 2(-deltadeltaCt) are 2.436 8, 4.4588 and 1.781 8 respectively. The increasing expression of NT-3 correlated positively with the scores of visual analog scale.@*CONCLUSION@#The expression of NGF, BDNF and NT-3 mRNA are as high as 2.4368, 4.4588 and 1.7818 times to healthy adults. We propose NGF, BDNF and NT-3 may contribute to the pathogenesis of AR. NT-3 could reflect the severity of rhinitis as a molecular biological index.
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Adolescente , Adulto , Niño , Femenino , Humanos , Masculino , Adulto Joven , Factor Neurotrófico Derivado del Encéfalo , Sangre , Genética , Factor de Crecimiento Nervioso , Sangre , Genética , Neurotrofina 3 , Sangre , Genética , ARN Mensajero , Genética , Rinitis Alérgica , SangreRESUMEN
Molecular biological methods were used to construct the eukaryotic expression vectors of precore and core gene named EBO PreC/C. Then T1862, A1896, A1899 and A1896+A1899 variants were constructed by site mutagenesis in vitro. By PCR RFLP and sequencing ,T1862,A1896,A1899 and A1896+A1899 variants were obtained. Wild type and variants were transfected to HepG2 cells, and HBeAg was tested to observe the difference of HBeAg expression between wild type and variants. After stable expression in HepG2 cells, HBeAg was detected to be positive in cells transfected wild type and A1899 variants, and negative in cells transfected with T1862, A1896,A1896+A1899 variants. The construction of these variants will play an important pole in studying the relation of PreC/C mutations and HBV expression and replication of HBV genome.
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<p><b>OBJECTIVE</b>To study the influence of host cellular HLA-I molecules expression by HBV precore region mutants.</p><p><b>METHODS</b>The plasmids of the wild type of HBV precore region and the mutants of A83 and A83/A86 were constructed, and then transected into HepG2 cells. The biological activity of HBV precore gene in the host cells was identified. The expression of HLA-I molecules was detected by flow cytometric analysis.</p><p><b>RESULTS</b>DNA segments similar with HBV precore gene size and HBeAg were detected by PCR and ELISA, respectively. The fluorescence intensity of HLA-I on host cells was different: wild type being 1.3; A83, 17.6; and A83/A86, 7.3.</p><p><b>CONCLUSIONS</b>HBV precore gene hot-spot mutants in vitro can increase the expression of HLA-I molecules on host cells.</p>
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Humanos , ADN Viral , Antígenos HLA , Alergia e Inmunología , Células Hep G2 , Antígenos e de la Hepatitis B , Virus de la Hepatitis B , Genética , Mutación , Reacción en Cadena de la PolimerasaRESUMEN
Objective To study HBeAg change in patients infected hepatitis B virus(HBV) with pre-C signal enzyme cleavage site mutation. Methods Mutation in pre-C signal enzyme cleavage site was detected by PCR-RFLP. The PreC/C gene with mutation was amplified by PCR and was cloned to EB viral eukarotic expression vector. Then transfect the vector with wild type or mutant PreC/C gene to HepG2 cell. SEAP reporter system was used to monitor the efficiency of transfection. HBeAg and its precursor in the supernatant and HepG2 cell were detected by ELISA and Western blot. Results HBeAg was positive in the supernatant of wild type and negative control in T1862 vaniant by ELISA. In HepG2 cell transfected with wild type, three proteins were detected by Western blot, they were HBeAg(17 000) and two HBeAg precursor(22 000 and 25 000). And in HepG2 cell transfected T1862 vaniant, only two HBeAg precursor was detected. The precursor in cells transfected withT1862 vaniant were significantly stronger than cells transfected with wild type. Conclusion Mutation in pre-C signal enzyme cleavage site may affect the decoration of HBeAg, which may cause great of HBeAg precursor locating in cells and lead to HBeAg negative in serum of patients infected with HBV.