Objective:To investigate the m echanism of D- amino acid oxidase/D- Alanine system in killing K5 6 2 e cells in vitro.Methods:The killing effects of D- Ala on K5 6 2 e cells stably expressing DAAO and GFP were observed.H2 O2 production by DAAO+ cells were m easured by the phenol red oxidation assay.L owry method was used to determ ine the protein quantities of cells and fluorescent intensities of GFP+ cells were assayed by flow cytom eter.Results:KDf Gd cells were killed completely after treated with 2 5 mm ol/L D- Ala for 2 4 h.The effect of D - Ala at 2 0 m mol/L on KDf Gd cells increased apparently within 4 8h,but the same effect was not observed if D - Ala was below 15 m mol/L .The cytotoxicity of D- Ala in KDf Gd cells was more sensitive than in parental K 5 6 2 e cells.The H2 O2 levels in the medium were consistent with the killing effects of D- Ala.Conclusion:The killing effects of DAAO/D- Ala system is m ediated by H2 O2 . [

OBJECTIVESTo investigate the relationship between the expression of mrp and both the responses to chemotherapy and the level of alpha-fetoprotein (AFP).

METHODSS-P immunohistochemical staining and in situ PCR were adopted to detect MRP and mRNA mrp in 54 cancer tissues taken from untreated HCC patients whose tumor could not be removed during the operation, 24 para-cancer tissues, and 12 posthepatitis cirrhosis paraffin-embedded tissues. The relationship between the expression of mrp and their curative effect to chemotherapy in all the patients was analyzed, so was the relationship between the expression of mrp and the level of AFP in 38 patients whose AFP was positive after operation.

RESULTSThe positive rates of expressing MRP and mRNA mrp in the three kinds of tissues were 61.1%, 25.0%, 33.3% and 77.8%, 37.5%, 41.7%, respectively, with higher rates in HCC tissues than those in other tissues (chi2=9.842, P< 0.01; chi2=13.956, P<0.01). The rates of curative effect to chemotherapy in groups of negative and positive MRP and mRNA mrp expression were 61.9%, 30.3% and 75.0%, 33.3%, respectively, with significant difference between the negative and positive groups (chi2=5.242, P<0.05; chi2=6.627, P< 0.05). As the same as the percentage of curative effect to chemotherapy, the rates of AFP level decreased evidently were 62.5%, 27.3% and 87.5%, 30.0%, with remarkable difference between the two groups (chi2=4.710, P<0.05; chi2=8.566, P<0.05).

CONCLUSIONSThe multidrug resistance (MDR) of HCC is related to mrp expression, which initiates the intrinsic MDR. There is an important significance by detecting mrp expression in selecting chemotherapeutic method, and the expression of mrp can act as an indicator for chemotherapeutic sensitivity in HCC patients.

Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; biosynthesis ; genetics ; Carcinoma, Hepatocellular ; chemistry ; drug therapy ; genetics ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; chemistry ; drug therapy ; genetics ; Male ; Middle Aged ; Multidrug Resistance-Associated Proteins ; biosynthesis ; genetics ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; alpha-Fetoproteins ; metabolism

BACKGROUNDIt is still controversial whether or not the correlation between lipid abnormality and coronary heart disease (CHD) becomes weaker in the elderly, and whether patients above 80 years old still benefit from lipid management for the secondary prevention of CHD. The purpose of this study is to assess the correlation between hyperlipidemia and the risk of CHD events in the elderly, and to determine if it is appropriate to use lipid-lowering drugs in those aged above 80, as prescribed by the recommended guidelines for lipid management.

METHODSOne thousand two hundred and eleven retirees, mainly males (92%), aged 70 +/- 9 years, were enrolled in this study. Lifestyle habits and medical history were recorded via questionnaires. During the period 1986 - 2000, all subjects participated in an annual physical examination with a blood chemistry survey. The mean follow-up period was 11.2 years. Subjects with incidental illnesses, especially cardiovascular diseases, were diagnosed or treated promptly. Serum lipid parameters, including total cholesterol (TC), low and high-density lipoprotein cholesterol (LDL-C and HDL-C) and triglyceride (TG) levels were analyzed according to standardization of lipid and lipoprotein measurements. The association between lipid levels and the prevalence of acute myocardial infarction (AMI) or coronary death was analyzed statistically.

RESULTSLipid abnormalities occurred in 2/3 of the 1211 subjects. The most common lipid disorder was high TC and high LDL-C, which was much more prevalent than high TG. Among the subjects, 51.6% had TC levels above 5.2 mmol/L. Mean TC and LDL-C reached peak levels in the 65 - 74 age group without significant decrease until ages over 90. The cumulative total number of deaths due to various causes was 397 in the 15-year follow-up period, with the mortality rate in the high lipid group slightly lower than that in the normal lipid group (30.6% vs 35.3%), although the difference was not significant (P = 0.1931). However, there were more cases of coronary death in the high lipid group than in the normal lipid group (7.9% vs 4.6%, P = 0.0045). When examining AMI survivors, more AMI cases were found in the high lipid group than in the low lipid group (20.9% vs 11.4%, P < 0.0001). The cumulative number of coronary deaths was 89 (with 88 cases above age 70), and the total number of CHD cases was 214 (17.7% of the whole group). Logistic regression analysis reveals that age, hypertension, LDL-C, and HDL-C are important risk factors for CHD. Lifestyle changes were common, but only 45% of the hyperlipidemic cases received drug treatment. Statins were commonly used only in recent years.

CONCLUSIONThe above results show that high TC and LDL-C levels are correlated with a high CHD risk even in people over 80. For elderly patients with clinical CHD and an aggregation of CHD risk factors, cholesterol-lowering therapy might be considered if the general health of the patient makes this permissible.

Aged ; Aged, 80 and over ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Coronary Disease ; blood ; etiology ; mortality ; Female ; Follow-Up Studies ; Humans ; Hyperlipidemias ; complications ; mortality ; Lipids ; blood ; Male ; Risk Factors ; Triglycerides ; blood

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 the effect of pulmonary surfactant (PS) on the Th1/Th2 balance and serum levels of interleukin-4 (IL-4), interferon-γ (IFN-γ) and IgE in neonates with respiratory distress syndrome (RDS).

METHODSA total of 58 neonates with RDS were divided into control (n=20) and PS treatment groups (n=38). The control group underwent mechanical ventilation and other conventional treatment, while the PS treatment group received with bovine PS treatment within 1 hour of being admitted to the hospital together with mechanical ventilation and other conventional treatment. Enzyme-linked immunosorbent assay was used to measure serum levels of IL-4, IFN-γ and IgE before treatment and 24, 48 and 72 hours after treatment. Simultaneously, arterial blood gas, respiratory system compliance, and other ventilator parameters were recorded.

RESULTSCompared with the control group, the PS treatment group showed significantly shorter duration of mechanical ventilation and oxygen exposure time (P<0.05), significantly better respiratory system compliance and significantly lower oxygenation index 24, 48 and 72 hours after treatment (P<0.05). At 48 and 72 hours after treatment, serum levels of IFN-γ were significantly lower in the PS treatment group than in the control group (120±46 ng/L vs 229±59 ng/L, P<0.05; 141±40 ng/L vs 282±44 ng/L, P<0.05), and serum levels of IL-4 were significantly higher in the PS treatment group than in the control group (263±48 pg/mL vs 152±45 pg/mL, P<0.05; 417±49 pg/mL vs 201±46 pg/mL, P<0.05). At 72 hours after treatment, serum level of IgE was significantly lower in the PS treatment group than in the control group (115±44 pg/mL vs 199±43 ng/mL; P<0.05).

CONCLUSIONSPS treatment can shorten the duration of mechanical ventilation and oxygen exposure time, regulate serum levels of IFN-γ, IL-4 and IgE, and influence Th1/Th2 balance in neonates with RDS, thus inhibiting lung inflammatory response and reducing lung injury.

CD4 Lymphocyte Count ; Female ; Humans ; Immunoglobulin E ; blood ; Infant, Newborn ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Pulmonary Surfactants ; pharmacology ; therapeutic use ; Respiration, Artificial ; Respiratory Distress Syndrome, Newborn ; complications ; drug therapy ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

BACKGROUNDThis study was designed to evaluate the relationship between high-density lipoprotein cholesterol (HDL-C) level and acute myocardial infarction (AMI) and coronary heart disease (CHD) death and to explore the protective effect of HDL against CHD in the elderly Chinese.

METHODSStarted from 1986, 1211 retirees (92% males) were enrolled consecutively and studied prospectively. The average starting age was 70 +/- 9 years, and that at the end of the study was 80 +/- 9 years. During the follow-up study, all the participants received yearly physical examination and blood chemistry survey from 1986 - 2000. The average duration of the follow up study was 11.2 years. The end point of this study was either attacks of AMI or death due to CHD and other causes. CHD risk factors were screened by logistic regression analysis. According to their HDL-C levels, cases were divided into low (< 1.03 mmol/L), medium (or normal, 1.03 - 1.56 mmol/L) and high (> 1.56 mmol/L) level groups, the differences in incidence of AMI and CHD death in each group were analyzed.

RESULTSThe cumulative attacks of acute coronary syndrome (mostly AMI) were 214 cases, including 89 cases of coronary death and 308 death caused by other diseases during the follow up study. AMI occurrence and CHD death in normal HDL-C group were lower than those in the low HDL-C group by 40% and 53%; and those in the high HDL-C group were lower than in the normal group by 56% and 50%, respectively. Statistical analysis on normal lipid cases (411 cases, total cholesterol < 5.17 mmol/L, triglyceride < 1.69 mmol/L) revealed that the cases at low HDL-C level had similar rates of AMI events and CHD mortality as those of the entire group (including hyperlipidemia); however, AMI attacks and CHD deaths decreased significantly at the normal and high HDL-C levels. The results demonstrated that the protective effect of HDL against coronary artery disease is more prominent in people with low lipid level.

CONCLUSIONLow HDL is an important independent risk factor for AMI attacks and CHD death in the elderly; high HDL has significant protective effect against coronary artery disease.

Aged ; Aged, 80 and over ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Coronary Disease ; blood ; etiology ; prevention & control ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; etiology ; Risk Factors

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

Age Distribution ; Aged ; Aged, 80 and over ; Calcium ; blood ; Female ; Herpes Zoster ; blood ; epidemiology ; Humans ; Incidence ; Male ; Middle Aged ; Neuralgia, Postherpetic ; blood ; epidemiology

Objective To explore the correlation between environmental selenium(Se) levels and cognitive ability among rural elderly population, and the effect of Se on cognitive skill of the elderly. Methods Two study sites(Zichuan district and Gaomi city of Shandong) with different environmental Se levels were selected according to rural elderly people ≥ 65 years were extracted by stratified random sampling method in each site. A retrospective survey was carried out using dietary intake questionnaire for the elderly for the past 1 year, and their daily total Se intake was calculated. Questionnaire was also used to obtain cognitive skill information in the elderly people. The Community Screening Instrument for Dementia, the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) Word List Learning Test, the Indiana University (IU) Story Recall Test, the Animal Fluency Test, and the IU Token Test were applied to assess the cognitive ability. Se level in these samples was analyzed. Correlation between the Se levels of environment and those of human body were analyzed statistically. Results There were 0.163), (0.405 ± 0.086)]mg/kg and nail Se[(147.44 ± 17.42), (117.38 ± 22.48)μg/L]between the two groups (U = 31.59, 25.00, 23.67, all P ＜ 0.01 ). There were positive correlation among the nail Se, environmental Se and Se in all subjects(r = 0.51,0.46, 0.60, all P ＜ 0.01 ). The differences of the CSID total score, the CERAD Word List Learning Test, the CERAD Word List Recall Test and the IU Story Recall Test between the two sites were statistically significant(F = 2.56, 9.18, 7.48, 4.42, all P ＜ 0.05), excluding the Indiana University Story Recall Test.After eliminating possible confounding factors, the Se levels and the CSID total score, the IU Story Recall Test, the Animal Fluency Test and the IU Story Token Test had a significantly positive correlation (r = 0.076, 0.138, 0.042,0.107, P ＜ 0.05 or ＜ 0.01 ), excluding the CERAD Word List Learning Test. Conclusions This study supports the hypothesis that a life long low Se level is associated with lower cognitive ability. The cognitive abilities in the elderly population lived in areas with high environmental selenium levels are significantly higher than that of the elderly lived in areas with low environmental selenium levels.

BACKGROUNDPulmonary hypertension (PH) frequently complicates the course of idiopathic pulmonary fibrosis (IPF) patients and is associated with significantly worse outcomes. The aim of the present study was to investigate the incidence of PH in IPF patients and evaluate the correlation between clinical parameters and systolic pulmonary artery pressure (sPAP).

METHODSHospitalized patients with IPF, who were evaluated for sPAP by Doppler echocardiography from January 2004 to December 2011, were enrolled in our study. Patients were defined as PH by an estimated sPAP > 50 mmHg and graded as PH likely, PH possible and PH unlikely, based on the 2009 European Society of Cardiology/European Respiratory Society PH Guidelines. The correlations between clinical parameters and sPAP were analyzed by multiple linear regression.

RESULTSTotally, 119 IPF patients were enrolled in our study and 28 (23.5%), 20 (16.8%) and 71 (59.7%) patients were PH likely, PH possible and PH unlikely, respectively. Borg dyspnea score was positively correlated with sPAP, r = 0.467, P < 0.001. Oxygen saturation was negatively correlated with sPAP, r = -0.416, P < 0.001. Diffusing capacity of the lung for carbon monoxide percentage predicted was negatively correlated with sPAP, r = -0.424, P = 0.003. N-terminal fragment of pro-brain natriuretic peptide and pulmonary artery width was positively correlated with sPAP, r = 0.452, P = 0.011 and r = 0.513, P < 0.001, respectively.

CONCLUSIONSThe incidence of PH in IPF patients was 23.5% in a single center of China. PH may worsen the dyspnea, right heart dysfunction and decrease the life quality of the patients with IPF.

Aged ; Echocardiography, Doppler ; Female ; Humans ; Hypertension, Pulmonary ; blood ; epidemiology ; physiopathology ; Idiopathic Pulmonary Fibrosis ; blood ; epidemiology ; physiopathology ; Incidence ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; blood ; Peptide Fragments ; blood ; Pulmonary Artery ; physiopathology ; Smoking