2.Construction of eukaryotic expression vector of hMTH1 gene antisense RNA.
Gao-feng JIANG ; Zhi-xiong ZHUANG ; Qi-zhan LIU ; Yun HE ; Liu-tao DU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(1):57-60
OBJECTIVETo construct pEGFP-C1-T vector, an eukaryotic expression plasmid of hMTH1 gene antisense RNA.
METHODSThe conservative region of hMTH1 gene was amplified by RT-PCR after total RNA being extracted from human embryo lung fibroblast (HLF) and then cloned into pGEM-T vector. After the recombinant plasmid was certified by DNA sequencing, the conservative region of hMTH1 gene was inserted into pEGFP-C1 vector reversedly and pEGFP-C1-T vector was constructed. The efficiency of antisense inhibition was verified by Western blotting after cell transfection.
RESULTS423 bp fragment including conservative region of hMTH1 gene was obtained by RT-PCR. After cloned by pGEM-T vector and certified by DNA sequencing, pEGFP-C1-T vector was successfully constructed by means of recombinant DNA technology. Additionally pEGFP-C1-T vector could efficiently decrease hMTH1 protein level by 46%.
CONCLUSIONThe efficient expression vector of hMTH1 gene antisense RNA, pEGFP-C1-T has been constructed successfully.
DNA Repair Enzymes ; Genetic Vectors ; genetics ; Humans ; Phosphoric Monoester Hydrolases ; genetics ; Plasmids ; RNA, Antisense ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction
3.Effects of hydroquinone on DNA and nucleus damage in human embryo lung fibroblasts.
Gao-feng JIANG ; Zhi-xiong ZHUANG ; Qi-zhan LIU ; Yun HE ; Liu-tao DU
Chinese Journal of Preventive Medicine 2003;37(3):183-185
OBJECTIVETo study DNA and nucleus damage in human embryo lung fibroblast (HLF) exposed to hydroquinone (HQ) and its genotoxicity.
METHODSHLF were treated with HQ (0, 10, 20, 40, 80 micro mol/L, respectively) for 3 h and DNA damage was detected by comet assay. HLF was also treated with the same concentrations of HQ for 1 h and micronucleus test was performed after they were cultured for 24 h.
RESULTSComet assay showed that percentage of cells with tails in each groups treated with varied doses of HQ was 12%, 19%, 42%, 79% and 95%, respectively, with mean tail length of 7.87, 9.35, 11.03, 19.28 and 23.32 micro m, respectively, in an obvious dose-dependent manner (P < 0.05). Very significant increase in percentage of cells with tails and length of their comet tail were observed in those groups treated with HQ of 20, 40 and 80 micro mol/L (P < 0.01). And, proportion of high and severe DNA damage increased with dose of HQ. HQ could also induce formation of micronucleus and abnormal nucleus in all groups treated by varied doses of HQ, with rates of micronucleus and abnormal nucleus of 2%, 3%, 10%, 9% and 15%, and 6%, 7%, 16%, 27% and 28%, respectively, in a significant dose-dependent manner. There was significant increase in rates of micronuclei and abnormal nuclei in cells treated with HQ at doses of 20, 40 and 80 micro mol/L (P < 0.05).
CONCLUSIONSExposure to HQ could cause DNA and nucleus damage inducing genotoxic effects on HLF.
Cell Nucleus ; drug effects ; Comet Assay ; DNA Damage ; drug effects ; Embryo, Mammalian ; Fibroblasts ; cytology ; Humans ; Hydroquinones ; toxicity ; Lung ; cytology ; Micronucleus Tests
4.Construction of double-strand break repair protein hKu70 deficient cell strain and its biologic characters.
Qi-zhan LIU ; Zhi-xiong ZHUANG ; Gao-feng JIANG ; Yun HE ; Lu-tao DU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(2):105-107
OBJECTIVETo construct DNA double-strand break (DSB) repair protein hKu70 deficient cell strain and to observe its biological characters for studying the functions of hKu70 gene and the effects of occupational harmfulness factors on DSB repair.
METHODSHuman lung fibroblasts (HLF) were transfected with the eukaryotic expression plasmids of hKu70 gene antisense RNA (pEGFP-C1-K) to construct hKu70 protein deficient cells (named as "HLFK"). The protein expression levels of hKu70 gene in HLFC and HLFK were detected by the Western blotting to estimate the effects of antisense inhibition. Morphology, growth character and growth status in soft agar of transfected HLFK were observed.
RESULTSpEGFP-C1-K vector was successfully expressed in HLF. The protein expression level of hKu70 gene in HLFK was decreased by 42% as compared with that in HLFC. No obvious changes of the biologic characters were observed in HLFK.
CONCLUSIONThe hKu70 protein deficient cell strain was successfully constructed. The hKu70 protein deficiency alone didn't induce obvious changes of the biological characters in HLFK.
Antigens, Nuclear ; analysis ; Cell Division ; DNA Damage ; DNA Helicases ; DNA Repair ; DNA-Binding Proteins ; analysis ; deficiency ; Humans ; Ku Autoantigen ; RNA, Antisense ; Transfection
5.The status on care and nutrition of 774 children staying in rural areas while parents were in towns
Xi-Chun PENG ; Jia-You LUO ; Kuan-Bao YAO ; Ru-Shan HU ; Qi-Yun DU ; Ming-Yuan ZHU
Chinese Journal of Epidemiology 2008;29(9):860-864
Objective To understand the status on care and nutrition of children living in the rural areas (so called 'left-behind' children) while their parents were seeking for jobs in the urban areas. Methods Cross-sectional study was employed in this investigation.The group of‘left-behind' children (n=774) and the comparison group (n=774) were identified.The ascertainment methods mainly included questionnaire,anthropometries measurements,food-frequency,and laboratory examination.Results be poorly attended (X2=6.671,P=0.036;X2=15.053,P=0.001 ).Mothers who chose to work outside of their households would tend to choose bottle-feeding or decrease the duration of breastfeeding for their infants (X2=5.051,P=0.031;t=-7.201,P=0.001).The intake of milk and bean products in children were obviously lower than that seen in the control group (t=-2.150,P=0.032 ;t=-2.054,significant difference when comparing with the control group (X2=4.560,P=0.033 ).Conclusion communities,parents and extended families would facilitate more attention and effective intervention programs to improve the situation.
6.Risk prediction model of perinatal congenital heart disease
Li-Bo ZHOU ; Ling ZHENG ; Jia-You LUO ; Qi-Yun DU ; Jun-Qun FANG ; Zhen-Qiu SUN
Chinese Journal of Epidemiology 2008;29(12):1251-1254
Through analyzing the influencing factors of congenital heart disease (CHD), it is aimed to establish CHD risk prediction model in fetus, and simultaneously provide theoretical foundation for CHD prevention. One-factor logistic regression method was used to screen the significant factors regarding CHD, and to separately adopt multiple-factor non-conditional logistic regression method and decision tree to set up model prediction fetus CHD risk and to analyze the advantages and shortcomings. Correct classification rates turned to be 80.93% and 82.79% respectively among 215 'training samples' by the two methods and the rates were 85.45 % and 89.09% respectively among 55 'testing samples'. The alliance of logistic regression and decision tree can overcome influence by co-linearity to guarantee the accuracy and perfection, as well as promoting the predictive accuracy.
7.Study on emotional, behavioral problems and related determinants among stranded children aged 4 to 7 years in the rural areas of China
Rong ZENG ; Ling-Li ZHANG ; Jia-You LUO ; Wen-Jie GONG ; Qi-Yun DU ; Hong WU
Chinese Journal of Epidemiology 2009;30(7):706-709
Objective To understand emotional, behavioral problems and related determinants among rural stranded children and to provide evidence for further psychological support. Methods A total of 3944 caregivers of children in the rural areas and aged 4 to 7 years but without parents around, were randomly selected. Data were collected through Strengths and Difficulties Questionnaires (the edition for parents, SDQ). Results The overall prevalence of children with behavioral/emotional problems was 43.6% among those stranded children including 8.3% having experienced emotional symptoms, 9.5% reported to be displaying 'conduct' problems, 8.7% exhibited significant hyperactivity/inattention, 18.9% experienced peer relationship problems and 16.8% having prosocial behaviors. The prevalence of behavioral/emotional problems was significantly higher in boys than those in girls and in lower age groups. Significant differences were also found in conduct problems, hyperactivity/inattention and prosocial behaviors (P<0.01). The result from logistic regression analysis showed that factors as education level and motivation of the caregivers as well as financial support from the parents were related to emotional/ behavioral problems among the stranded children. Conclusion The prevalence of emotional and behavioral problems was high among children living the rural areas but without their parents around which should call for psychological intervention.
8.A case-control study on genetic and environmental factors regarding polydactyly and syndactyly
Jia-You LUO ; Chu-Hui FU ; Kuan-Bao YAO ; Ru-Shan HU ; Qi-Yun DU ; Zhi-Yu LIU
Chinese Journal of Epidemiology 2009;30(9):903-906
king condition before pregnancy were associated with the development of polydactyly and syndactyly.
9.Odds ratio on cardiovascular risk factors of obesity defined by waist and body mass index
Song-Ming DU ; Yan-Ping LI ; Hong-Yun FANG ; Xiao-Qi HU ; Xiao-Guang YANG ; Guan-Sheng MA ; Yong-Hua HU
Chinese Journal of Epidemiology 2010;31(6):626-632
Objective To compare the odds ratio of waist circumference (WC) and/or body mass index (BMI) on cardiovascular risk factors. Methods Data on a cross-sectional study involving 41 087 adults (19 567 male, 21 520 female) from the 2002 China National Nutrition and Health Survey were examined. According to the obesity definition of the Chinese Working Group on Obesity for Children (WGOC) (BMI, 24 and 28 kg/m2; WC, male 85 cm, female 80 cm), the study population were divided into 9 groups. The prevalence and odds ratio (OR) of cardiovascular disease (CVD) risk factors were compared among these 9 groups. Variation and standard β were indexes being used to compare the likelihood of BMI and/or WC on CVD risk factors. Results Within each of the BMI categories, with few exceptions, indices levels on CVD risk factors were significantly increased (decreased for HDL-C levels) with the increase of WC, and vice versa. After adjusting the effects of age, sex, income, education, sedentary activity and dietary factors, the ORs of hypertension in adults with higher WC within each of the BMI categories were higher than adults with lower WC, and the ORs of hypertension in adults with higher BMI within each of the WC categories were higher than adults with lower BMI. Similar trends were found for high fast plasma glucose (FPG) and dyslipidemia. The variation in CVD risk factors explained only by WC and BMI were quite similar,but slightly larger when combined WC with BMI. Standard β was higher on BMI when predicting systolic BP and was higher on WC when predicting TG, TC and HDL-C. Conclusion BMI and WC were independently associated with the risk factors on CVD.To combine the BMI and WC, the results could accurately evaluate the risk of CVD, thus to provide substantive evidence that the WGOC cutoff points for the WC might help in identifying those population under increased risk.
10.Extraction and Separation of Boron in Anhydrite and Gypsum Minerals and Its Isotopic Measurement by Thermal Ionization Mass Spectrometry
Jie Zhan QIN ; Ru Xiang ZHANG ; Kuang Zhang PENG ; Kuan Qing LI ; Qi Yun MA ; Shun Qi FAN ; Sheng Yong DU ; Ping Jian WANG ; Shou Fa SHAN
Chinese Journal of Analytical Chemistry 2018;46(1):48-54
The anhydrite and gypsum are the main sulfate minerals during evaporation of seawater or lake.They record the information about relative hydrogeology and the composition of mother liquor.Boron is diffluent element, and often occurs in all kinds of evaporites.Presently, the boron isotope has been applied widely in mineral deposits forming, geochemistry and palaeoenvironment.However, there is little research about characteristic of boron isotope in anhydrite and gypsum minerals, because of the low content of boron and micro-solubility in water and hydrochloric acid.This study developed a method of extracting and purifying boron in anhydrite and gypsum by phase transformation and ion-exchange.Firstly, the samples were mixed with ammonium hydrogen carbonate to transform the calcium sulfate to calcium carbonate.And diluted hydrochloric acid (1 mol/L) was added to resolve calcium carbonate.The percent conversion was about 85%in the first stage, and up to complete resolution by repeating this process.Secondly, boron specific ion-exchange resin ( Amberlite IRA 743 ) was used to gather the boron ions fully and further refined the samples with more than 1 μg of boron by anionic and cationic resin mixed by Ion Exchange Ⅱ and Dowex 50 W × 8.Finally, according to the modified method by He, the values of boron isotope were determined by TIMS.The boron content is analytically pure gypsum was 3.501 ± 0.128 μg/g ( n=12 , RSD=3.6%) and the average recovery was 100.47%.Besides, the δ11B value of analytically pure gypsum added with NIST SRM 951 was 17.98‰±0.21‰ (n=3, RSD=1.2%).This method has good repeatability and can meet the requirements of boron isotopic measurement of anhydrite and gypsum.