Objective:To investigate the in vitro effect of capsaicin on four major rat cytochrome P450 ( CYP) enzymes using a cocktail probe drug method. Methods:The in vitro incubation was divided into capsaicin group and the control group. Rat liver micro-somes, probe drugs, capsaicin at various concentration ( buffer solution in the control group) and cofactors were cultured altogether for 20 min. After the culture, the concentration of metabolites was determined by HPLC to assess the activities of enzymes. IC50 value of capsaicin on every isoform was calculated using Graphpad prism 5. 0. Capsaicin and hepatic microsomess were pre-incubated respec-tively for 0, 5, 10, 15, 20 and 30 min, and then the relative activity of the four isoforms at different time was calculated. Results:The activity of the rat liver microsomes enzyme CYP450 isoforms (CYP1A2, CYP2C11, CYP2E1 and CYP3A2) was all inhibited by capsaicin in vitro with IC50 value of 36. 21, 17. 19, 51. 64 and 18. 86 μmol·L-1 , respectively. Pre-incubation could not increase cap-saicin inhibitory activity against the four CYP enzymes. Conclusion:Capsaicin shows inhibition on CYP1A2, CYP2C11, CYP2E1 and CYP3A2 in rat liver microsomes in vitro without pre-incubation time-dependent property.

The internal ribosome entry site (IRES) sequence was derived from encephalomyocarditis virus. It allows to translate two open reading frames at one mRNA, so two genes conjoined by IRES have the same expression rate. K(DfGC) and K(DfGd) cell lines, stably expressing D-amino acid oxidase (DAAO) gene and green fluorescence protein (GFP) genes, were obtained by transfection of K562e cells with retroviral vector pLDfG containing IRES sequence, DAAO cDNA and GFP gene. Fluorescence positive rate and fluorescence intensity of the two cell lines were measured with flow cytometry. H(2)O(2) production by K(DfGC) and K(DfGd) cells treated with D-alanine was measured by the phenol red oxidation assay. The fluorescence positive rate and fluorescence intensity in K(DfGC) and K(DfGd) cell were 94.64% and 96.31% and 202 units and 174 units per 2 x 10(4) cells, respectively. There was exponential correlation between fluorescence intensity and H(2)O(2) level. The above-mentioned results demonstrate that DAAO gene and GFP gene were simultaneously expressed in K562e cell line by the regulation of IRES sequence, and DAAO level was correlated with fluorescence intensity of GFP.
Binding Sites ; genetics ; D-Amino-Acid Oxidase ; genetics ; metabolism ; Gene Expression ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; Humans ; Hydrogen Peroxide ; metabolism ; K562 Cells ; Luminescent Proteins ; genetics ; metabolism ; Retroviridae ; genetics ; Ribosomes ; metabolism ; Transfection

OBJECTIVETo investigate mesenchymal stem cells (MSCs) immunosuppressive activity in the presence of interferon-gamma (IFN-γ) to reveal synergistic immunomodulatory effects of IFN-γ and MSCs.

METHODS① MSCs were cultured in the presence or absence of IFN-γ（100 ng/ml）, the supernatants were collected for measurements of PGE2、HGF and TGF-β1 by ELISA kits. ② MSCs were cultured in the presence or absence of IFN-γ （100 ng/ml）for 48 h. The cDNA was analysed for the expression of human indoleamine 2, 3-dioxygenase（IDO）mRNA by semiquantitative RT-PCR. ③ Mononuclear cells (MNCs) were extracted from peripheral blood of healthy donors. The T cell proliferation was tested in the co-culture system added with MSCs, recombinant human IFN-γ (100 ng/ml) and anti-IFN-γ mAb (5 μg/ml) by BrdU ELISA kit.

RESULTS①The immunosuppressive cytokines PGE2、HGF and TGF-β1 were detectable within 24-48 h in the supernatants. Their expressions were significantly up-regulated in the presence of IFN-γ. Concentrations of these cytokines were as of (1715.5±628.6) pg/ml vs (1344.5±709.4) pg/ml (P=0.001);(4031.8±1496.8) pg/ml vs (2452.4±1375.3) pg/ml（P=0.011）;(1753.5±413.8) pg/ml vs (1026.6±450.5) pg/ml（P<0.001）,respectively. ②The expression of IDO mRNA was undetectable when MSCs were cultured alone. In contrast, The IDO mRNA expression was remarkably enhanced in the presence of IFN-γ. ③Bone marrow-derived MSCs remarkably suppressed allogeneic T cell proliferation in vitro. Addition of exogenous IFN-γ had no significant effect on the inhibitory capacity of MSCs, the inhibitory ratios of T cell proliferation were （40.4±10.9）% vs（36.7±7.4）% (P=0.272). By contrast, the inhibitory ratio of T cell proliferation was significantly decreased in the presence of anti-IFN-γ mAb[(40.4±10.9)% vs (23.9±7.6)%,P=0.002].

CONCLUSION①Human MSCs constitutively expressed immunosuppressive concentrations of PGE2, HGF and TGF-β1, and their expressions were significantly up-regulated by IFN-γ. ②IFN-γ-induced expression of IDO on MSCs involved in tryptophan catabolism. ③MSCs notably suppressed allogeneic T cell proliferation in vitro. IFN-γ promoted the immunosuppressive capacity of human MSCs, indicating the synergistic immunomodulatory effect of IFN-γ and MSCs.

Bone Marrow Cells ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytokines ; immunology ; Humans ; Immune Tolerance ; Interferon-gamma ; pharmacology ; Mesenchymal Stromal Cells ; immunology ; T-Lymphocytes ; cytology

OBJECTIVETo observe blood lead level and related risk factors among children of 0 - 6-year old in Beijing.

METHODSStratified-clustered-random sampling and simple random sampling were used. A total of 2 262 children of 0 - 6 years old were investigated from May to July 2001. They were permanent residents in Beijing. Blood lead level was tested by graphite atomizer absorption spectrophotometer. At the same time, related factors were investigated using a standardized questionnaire.

RESULTSThe mean lead level of children in Beijing was 96.8 micro g/L with 35.7% of those >/= 100 micro g/L, and 2.5% >/= 200 micro g/L. The proportion of lead poisoning was significantly higher in rural children (P < 0.01). The proportion of lead poisoning in 2-year-old group was higher than that in other age groups. The proportion of lead poisoning in boys was significantly higher than that in girls (P < 0.01). The following factors might serve as major risk factors related to child lead poisoning, such as, never or rarely drinking milk, living in nearby highways (less than 50 meters) or living in the first floor/bungalow, and so on.

CONCLUSIONThe blood lead level of >/= 100 micro g/L among Beijing children appeared to be a big problem. Decision-makers should pay more attention to prevent blood lead level being high, and to cure these children who suffered in lead poisoning. Effective intervention measures on these target populations should be taken.

Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Lead ; blood ; Lead Poisoning ; therapy ; Male ; Risk Factors ; Sex Factors

OBJECTIVETo explore the influence of compound whole grain complex antioxidant chain on oxidative stress to the hyperlipidemia population.

METHODSFrom March 2008 to March 2009, 418 hyperlipemia residents (45 to 75 years of age) of Han group were screened from 3 main districts in Nanjing, according to the community, blood lipids and oxidative indicators, stratified into intervention and control group by quasi-experimental design. The intervention group (212 individuals) were provided with compound whole grain and health education while only health education was provided for the control group (206 individuals). Body mass index (BMI), waist-to-hip ratio (WHR), total cholesterol (TC), triglyceride (TG) and high density lipoprotein cholesterol (HDL-C) and oxidative indicators (including T-AOC, MDA, SOD, GSH-Px) were measured before and after the one-year intervention period. Analyses of the correlation between posture, biochemical markers and oxidative stress indicators before and after intervention were carried out.

RESULTSAfter intervention, BMI ((25.53 ± 2.77) kg/m(2)), WHR (0.82 ± 0.03), TC ((4.60 ± 0.98) mmol/L), TG ((1.26 ± 0.88) mmol/L) in the intervention group were decreased significantly compared to the levels of BMI ((26.60 ± 3.18) kg/m(2)), WHR (0.93 ± 0.05), TC ((4.97 ± 1.02) mmol/L), TG ((1.98 ± 1.11) mmol/L) in the control group (all P values < 0.05); while HDL-C ((1.34 ± 0.26) mmol/L) in the intervention group was increased significantly compared to the level of HDL-C ((1.18 ± 0.17) mmol/L) in the control group (P < 0.05); After intervention, levels of T-AOC (19.52 ± 0.81), SOD ((85.42 ± 21.65) U/ml) and GSH-Px ((128.26 ± 33.65) µmol/L) were increased significantly compared to the levels of T-AOC (11.11 ± 1.30), SOD ((78.68 ± 30.48) U/ml) and GSH-Px ((118.48 ± 24.19) µmol/L) in the control group (all P values < 0.05); while MDA ((1.78 ± 1.16) nmol/ml) decreased significantly compared to the level of MDA ((2.12 ± 1.37) nmol/ml in the control group (P < 0.05); Pearson product moment correlation analysis showed that: T-AOC with TC, TG, BMI showed a negative correlation (r values were -0.258, -0.266, -0.230, respectively, all P values < 0.05), while with HDL-C was a positive correlation (r values was 0.194, P < 0.05); SOD with TC, TG, BMI showed a negative correlation (r values were -0.282, -0.311, -0.217, respectively, all P values < 0.05), while with HDL-C was a positive correlation (r values was 0.169, P < 0.05).

CONCLUSIONCompound whole grain could improve lipid metabolism to the hyperlipidemia population. There was a correlation between common human metabolism and the levels of oxidative stress.

Aged ; Aged, 80 and over ; Antioxidants ; China ; epidemiology ; Diet ; Diet Surveys ; Edible Grain ; Female ; Health Education ; Humans ; Hyperlipidemias ; epidemiology ; metabolism ; prevention & control ; Lipid Metabolism ; Male ; Middle Aged ; Oxidative Stress

OBJECTIVETo explore the comprehensive program of integrated Chinese and western medicine in the treatment of cognitive impairment in earthquake brain injury.

METHODSThe multi-central randomized controlled trial was adopted. The qualified subjects were randomized into an acupuncture + rehabilitation group (38 cases) and a rehabilitation group (35 cases). In the acupuncture + rehabilitation group, acupuncture, hyperbaric oxygen (HBO) and cognitive rehabilitation training were combined as the comprehensive program of integrated Chinese and western medicine in the treatment. In the rehabilitation group, HBO and cognitive rehabilitation training were adopted. The efficacy and safety were assessed.

RESULTS(1) After treatment of 2 months, the intelligent state, cognitive function and activity of daily life of patients were improved in the both groups (all P < 0.01). (2) After treatment of 2 months, the score of MMSE and the score of activity of daily life were (24.11 +/- 4.08) and (75.45 +/- 13.95) in the acupuncture + rehabilitation group, which were more significant as compared with (17.05 +/- 43.84), (66.06 +/- 12.75) in the rehabilitation group, respectively (both P < 0.01). In 6-month follow-up visit after treatment, the cognitive function and activity of daily life were improved continuously in the acupuncture + rehabilitation group, which was more significant as compared with the rehabilitation group (P < 0.01, P < 0.05).

CONCLUSIONThe integrated Chinese and western medicine of acupuncture, HBO and cognitive rehabilitation training is safe and effective in the treatment of cognitive impairment in earthquake brain injury. The therapeutic effect is more advantageous as compared with the simple rehabilitation program of western medicine.

Activities of Daily Living ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Brain Injuries ; psychology ; rehabilitation ; therapy ; Cognition ; Earthquakes ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Objective: To investigate the effects of donor-specific HLA antibodies(DSA) for graft failure in un-manipulated haploidentical hematopoietic stem cell transplantation(haplo-HSCT) and the feasible treatment for DSA. Methods: HLA antibodies were examined using the Luminex-based single Ag assay for 92 patients who were going on haplo-SCT and the correlations of graft failure and DSA among the patients who had finished SCT were analyzed. Results: Of the total 92 patients who were going on haplo-HSCT, sixteen (17.4%) patients were HLA Ab-positive, including six (6.5%) patients with antibodies corresponding to donor HLA Ags (DSA-positive). Among the patients who had finished the haplo-HSCT with conventional myeloablative conditioning regimen, the engraftment rate was significantly higher in DSA (-) patients than that in DSA (+) patients [92.3% (24/26) vs 25.0%(1/4), χ²=8.433, P=0.004] and DSA was the only factor relevant with graft failure in multiple-factor analysis [OR=12.0(95% CI 1.39-103.5), P=0.024]. Strategies to decrease antibody levels were taken for 4 patients, two were their first transplantations, and the other two patients were their second haplo-HSCT. Three of the four patients were HLA-I-DSA positive and had gained donor engraftment by means of donor platelet transfusions to decreased the level of DSA, the fourth patient with both HLA-I and HLA-II DSA also gained engraftment with the treatments of TBI, rituximab and donor platelet transfusion. Conclusion: DSA is one of the key factors of graft failure in haplo-HSCT. Donors should be selected on the basis of an evaluation of HLA antibodies before transplantation. If haplo-HSCT from donors with DSA must be performed, then recipients should be treated for DSA to improve the chances of successful engraftment.
Antibodies ; Graft vs Host Disease ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; Humans ; Tissue Donors ; Transplantation Conditioning

BACKGROUND AND PURPOSE: Epidemiological data of cerebral small vessel disease (CSVD) in the general population of China are lacking. We report on the prevalence of lacunes, white matter hyperintensity (WMH), and cerebral microbleeds (CMBs) in a community-based sample in China and compare the results with those of other studies. METHODS: This was a cross-sectional analysis of the population-based Shunyi Study in China. A total of 1,211 stroke-free participants (mean age, 55.6±9.3 years; 37.4% men) with available 3 Tesla (3T) magnetic resonance images were included in this analysis. Demographic information and risk factor data were assessed. The overall and age-specific prevalence of lacunes, WMH, and CMBs was evaluated. Associations between cardiovascular risk factors and the presence of these lesions were analyzed by multiple logistic regression. RESULTS: Our study showed a prevalence of 14.5% for lacunes, 72.1% for periventricular hyperintensity (PVH), 65.4% for deep white matter hyperintensity (DWMH), and 10.6% for CMBs. When compared with other community-based samples, individuals in the same age group showed a higher burden of lacunes and a relatively lower prevalence of CMBs. Advanced age was independently associated with the prevalence of these CSVD markers, while the presence of hypertension increased the risk of lacunes, PVH/DWMH, and CMBs in deep or infratentorial locations. CONCLUSIONS: A higher burden of lacunes but a relatively lower prevalence of CMBs was observed in this Chinese population. This notable result highlights the challenge of CSVD prevention in China. Chinese have a risk factor profile for CSVD similar to those in other populations.
Asian Continental Ancestry Group* ; Cerebral Small Vessel Diseases* ; China ; Cross-Sectional Studies ; Humans ; Hypertension ; Logistic Models ; Prevalence* ; Risk Factors* ; White Matter

3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics* ; Child, Preschool ; DNA Mutational Analysis ; Disorder of Sex Development, 46,XY/genetics* ; Female ; Humans ; Membrane Proteins/genetics* ; Mutation

OBJECTIVE: To investigate the effects of nuclear transcription factor-κB(NF-κB)/amide adenine dinucleotide phosphate oxidase 1(NOX1) signaling pathway in tumor necrosis factor-α(TNF-α) induced apoptosis of A549 cells. METHODS: i) A549 cells were stimulated with TNF-α at the concentrations of 0.00, 0.25, 0.50, and 1.00 nmol/L. CCK-8 assay was used to detect the cell viability to screen the optimal stimulating concentration of TNF-α. ii) A549 cells at logarithmic growth stage were randomly divided into four groups, the control group, the TNF-α group, the BAY11-7082(NF-κB inhibitor) group and the TNF-α+BAY11-7082 group. The cells in the control group were not treated. The TNF-α and BAY11-7082 groups were stimulated with 0.50 nmol/L TNF-α and 5 μmol/L BAY11-7082, respectively. The TNF-α+BAY11-7082 group was stimulated by both TNF-α and BAY11-7082. After 24 hours of culture, the cell survival rate was detected by CCK-8 assay. Flow cytometry was used to detect cell apoptotic rate, and Western blotting was used to detect the relative expression of NF-κB(p65) and NOX1 proteins. RESULTS: i) When A549 cells were stimulated with TNF-α at the concentration of 0.50 nmol/L, the cell proliferative activity was reduced and the cell apoptosis was promoted. This concentration was selected as the stimulation dose of TNF-α in subsequent experiments. ii) The survival rate of A549 cells in the TNF-α group decreased(P<0.05), the apoptotic rate and the protein expressions of NF-κB(p65) and NOX1 increased in TNF-α group(all P<0.05) compared with the control group. In BAY11-7082 group, the survival rate and the relative expression of NF-κB(p65) and NOX1 of A549 cells were decreased(all P<0.05), and the apoptotic rate of A549 cells was increased(P<0.05) compared with the control group. A549 cells in TNF-α+BAY11-7082 group changed from a long spindle shape to an irregular one. The cell survival rate increased(P<0.05), the apoptotic rate and the relative expression of NF-κB(p65) and NOX1 decreased(all P<0.05) compared with the TNF-α group. CONCLUSION: NF-κB/NOX1 signaling pathway is involved in A549 cells apoptosis induced by TNF-α.