1.Role of stanniocalcin 1 in brain injury of coal-burning-borne fluorosis rats
Xu-yi, CHEN ; Bing, LIANG ; Feng-wu, TANG ; Yong-chun, ZHANG ; Fa, SUN ; Jiang, GU ; Sai, ZHANG
Chinese Journal of Endemiology 2013;(2):129-132
Objective To observe the change of stanniocalcin 1 (STC1) and calcium content in brain of coal-burning-borne fluorosis rats,and to explore the role of STC1 in brain injury of coal-burning-borne fluorosis.Methods Twenty four male SD rats were randomly divided into control,low,medium,and high fluoride groups according to body mass.Control group was fed conventional rat chow(fluorinated 1.3 mg/kg),and low,medium and high fluoride groups fed with fluorinated feed(20.0,40.0,60.0 mg/kg).All rats were given distilled water and feed ad libitum.One hundred and eighty days after modeling,STC1 protein and gene expression in the brain tissue of rats were detected using immunohistochemistry and RT-PCR and calcium content of brain tissue was detected.Results The cell positive rates of STC1 in low,medium,high fluoride groups [(48.10 + 2.11)%,(54.90 ± 1.73)%,(79.30 ± 3.71)%] were significantly higher than that of the control group[(24.70 + 3.53)%,all P < 0.05],the cell positive rate of high fluoride group was significantly higher than that of the low and medium fluoride groups (all P < 0.05).The STC1 mRNA expression of low,medium and high fluoride groups (0.58 ± 0.09,0.85 ± 0.17,1.75 ± 0.04) were significantly higher than that in the control group(0.37 ± 0.12,all P< 0.05),the STC1 mRNA expressions of high fluoride group was significantly higher than that of the low and medium fluoride groups (all P < 0.05).The brain cortex calcium ion concentrations of low,medium and high fluoride groups[(138.62 + 4.19),(167.43 + 6.57),(189.45 + 3.72)nmol/L] were significantly higher than that in the control group [(101.47 + 9.46)nmol/L,all P < 0.05],the brain cortex calcium ion concentrations of high fluoride group was significantly higher than that of the low and medium fluoride groups(all P < 0.05),and the medium fluoride groups was higher than the low groups (P < 0.05).Conclusion STC 1 may be involved in brain damage of coal-burning-borne fluorosis rats through regulating calcium balance.
3.Development of an interferon-gamma ELISPOT for bovine tuberculosis.
Zhengzhong XU ; Fa SHAN ; Fengli SHAN ; Chuang MENG ; Xiaoli XIE ; Jiaying LIU ; Jingjing MIN ; Xiang CHEN ; Xin'an JIAO
Chinese Journal of Biotechnology 2015;31(2):183-194
We established an ELISPOT for bovine interferon-gamma (BoIFN-γ), and applied it in the diagnosis of bovine tuberculosis (bTB). Monoclonal antibodies that can bind with native BoIFN-γ were screened as the coating antibody and detecting antibody. After optimization of detecting conditions including coating antibody concentration, cell number, and detecting antibody concentration, the ELISPOT assay was established. Peripheral mononuclear cells (PBMCs) isolated from 30 cows were co-cultured with PPD, and detected with the ELISPOT assay. The optimal conditions of ELISPOT assay were 2.5 μg/mL coating antibody 2G5, 2.5 x 10(5) cells/well, and 1 μg/mL detecting antibody Bio-5E11. In these 30 cows tested both with the ELISPOT assay and the BOVIGAM kit, 11 cows were proved to be positive in ELISOPT assay with the sensitivity of 78.6%, and 12 cows were proved to be negative in ELISOPT assay with the specificity of 75%. The ELISPOT assay for BoIFN-γ could be used to detect bTB efficiently and it might be an alternative method for the diagnosis of bTB.
Animals
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Antibodies, Monoclonal
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Cattle
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Enzyme-Linked Immunospot Assay
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veterinary
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Female
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Interferon-gamma
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isolation & purification
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Sensitivity and Specificity
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Tuberculosis, Bovine
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diagnosis
4.Clinical analysis of organic acidemia in neonates from neonatal intensive care units.
Fa-Lin XU ; Tian FAN ; Jia-Jia DUAN ; Dan CHEN
Chinese Journal of Contemporary Pediatrics 2012;14(5):336-339
OBJECTIVETo study the clinical features of organic acidemia in neonates admitted to the intensive care unit.
METHODSThe clinical features of neonates from 15 neonatal intensive care units of Henan Province, who were diagnosed with congenital organic acidemia by gaschromatography-mass spectrometry (GC-MS) between June 2008 and August 2011 were retrospectively reviewed.
RESULTSFifty neonates of 287 high risk neonates were confirmed as having or highly suspected to have inborn errors of metabolism. Of the 50 cases, 32 cases were diagnosed with organic acidemia disease, including 28 cases of methylmalonic acidemia, 2 cases of propionic acidemia, 1 case of maple syrup urine disease and 1 case of isovaleric acldemla. In most cases, disease onset occurred in the first week after birth in most of cases (75%). Neonates whose symptoms occurred immediately after or within a few hours of birth presented with serious conditions. Clinical manifestations were various and mainly related to neurologic, respiratory and gastrointestinal symptoms such as poor response, coma, drowsiness, abnormal muscle tone, convulsions, polypnea, dyspnea, milk refusal, diarrhea and jaundice. Initial symptoms were non-specific and included dyspnea, poor response, milk refusal, lethargy and seizures.
CONCLUSIONSMethylmalonic acidemia is a common inherited metabolic disease in the neonatal period. Clinical manifestations of organic acid metabolism abnormalities in neonates are atypical and early onset is associated with more serious conditions.
Amino Acid Metabolism, Inborn Errors ; complications ; diagnosis ; Diagnosis, Differential ; Female ; Gas Chromatography-Mass Spectrometry ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal ; Male ; Maple Syrup Urine Disease ; complications ; diagnosis ; Propionic Acidemia ; complications ; diagnosis
5.Effects of long-term excessive Iodine intake on the maternal-fetal thyroid hormone metabolism in mice
Xue-Feng YANG ; Jian XU ; Huai-Lan GUO ; Xiao-Hui HOU ; Xiao-Yi CHEN ; Xiu-Fa SUN
Chinese Journal of Endocrinology and Metabolism 2000;0(06):-
Long-term excessive iodine intake resulted in an increased TT_4 level and a decreased TT_3 level in maternal serum,meanwhile,hepatic and renal type 1 deiodinase activity decreased dose-dependently.A significant reduction in type 2 deiodinase ( D2 ) activity of 12.5 d placenta was found in 3.0 mg/L or above groups.For 19.5 d uterus,D2 activity decreased and type 3 deiodinase activity increased.The results suggest that excessive iodine has an effect on the embryonic development by regulating maternal-fetal thyroid hormone metabolism.
6.Association of angiotensin converting enzyme gene polymorphism with genetic susceptibility to systemic lupus erythematosus
Yong WANG ; Jian-Hua XU ; Dong-Qing YE ; Fa-Ming PAN ; Ke-Chun ZHANG ; Li LIAN ; Hong CHEN ;
Chinese Journal of Rheumatology 2003;0(07):-
Objective The aim of this study is to investigate the role of angiotensin-converting enzyme (ACE)gene susceptibility to systemic lupus erythematosus(SLE)by familial studies.Methods PCR-based re- striction fragment length polymorphism(RFLP)was applied to genotype single nucleotide polymorphism(SNP) G261T of the ACE gene.A total of 119 patients with SLE from 119 families were recruited.In addition,316 family members of these patients were also genotyped.A family-based association study was carried out to ex- plore the association between gene polymorphism and SLE.We studied the SNP encoding non-synonymous substitution in the ACE gene with respect to genetic susceptibility to SLE.Results Among 119 SLE patients. the frequency of ACEG261TG,T alleles was 44.8%.55.2% respectively,the frequency of ACEG261T GG,GT and TT genotypes was 13.9%,62.0%,24.1% respectively,Univariate(single-marker)family-based association test(FBAT)demonstrated that variant alleles at the SNP,rs4303,exon 5 of ACE gene were significantly asso- ciated with genetic susceptibility to SLE in Additive Model(Z=2.877,P=0.004),Dominant Model(Z=2.557, P=0.011).Recessive Model(Z=2.202,P=0.028).Transmission-disequilibrium test(TDT)and sib transmission -disequilibrium test(STDT)showed an excess of the allele of T from heterozygous parents to affected offspring or higher frequency of the allele of T in the patients than their normal siblings(X~2=11.66,P=0.001).Conclu- sion Our findings suggest that the ACE gene may he the susceptible gene to SLE in Chinese population,and the individuals carrying ACE-261T allele is significantly associated with susceptibility to SLE.
7.Research on the behavior of fruit and vegetable intake in adolescents with Transtheoretical Model.
Chen-Jia MAO ; Liang-Wen XU ; Xu-Ping QU ; Qi-Fa YANG ; Han-Qiong HU ; Dong-Ming XU
Chinese Journal of Preventive Medicine 2010;44(5):444-447
OBJECTIVETo study the current situation and factors influencing the behavior of fruit and vegetable intake in adolescents, and to discuss health education strategy related to the behavior.
METHODS1677 students were selected from primary school and middle school in urban area of Hangzhou by a multistage sampling method. The behavior of fruit and vegetable intake was surveyed by questionnaires of the patient-centered assessment and counseling for exercise plus nutrition project (PACE+) for adolescent-scale of fruit and vegetable consumption, and analyzed by Transtheoretical Model on the servings of intake, the stage of change, process of change, decisional balance and the current status of self-efficacy.
RESULTSThe average intake of fruit and vegetable in adolescents was (3.21 +/- 1.50) servings per day ("one serving" means 100 g cooked vegetable or 100 g fruit). The behavior of consuming fruit and vegetable in most of the adolescents was in the contemplation stage, accounting for 36.91% (619/1677). The process of behavior change, decisional balance (Pros) and self-efficacy existed a positive correlation with the process of stage (r(process) = 0.38, r(decisional balance (Pros)) = 0.26, r(self-efficacy) = 0.33, t values were 16.78, 11.02 and 14.31, P < 0.05). The servings of fruit and vegetable intake existed a positive correlation with stage transition and self-efficacy (beta(stage transition) = 0.665, t = 35.07, P < 0.05; beta(self-efficacy) = 0.050, t = 2.63, P < 0.05), and existed a negative correlation with decisional balance (Cons) (beta(decisional balance (Cons)) = -0.051, t = -2.84, P < 0.05).
CONCLUSIONFruit and vegetable intake of these adolescents are under the recommended dietary intake. Along with behavior stage development and self-efficacy improvement, the intake of fruit and vegetable also increased correspondingly. Intervention strategies should aim at improving the awareness of adolescents on the health benefits of fruit and vegetable intake and promoting their confidence to reach the recommended servings.
Adolescent ; Adolescent Nutritional Physiological Phenomena ; Child ; China ; Eating ; Feeding Behavior ; Female ; Fruit ; Humans ; Male ; Models, Theoretical ; Students ; Surveys and Questionnaires ; Vegetables
8.Effect of tetrandrine on the doxorubicin-induced expression of mdr1 gene in K562 cells.
Xu-jing LU ; Wen-lin XU ; Wen-juan LUO ; Fa-chun WANG ; Qiao-yun CHEN
Chinese Journal of Hematology 2008;29(7):468-471
OBJECTIVETo investigate the effect of tetrandrine (TTD) on doxorubicin-induced mdr1 gene expression and its mechanism.
METHODSMTT assay was used to detect the cytotoxicity of TTD to K562 cells. K562 cells were treated with doxorubicin alone or 0.6 microg/ml doxorubicin combined with various concentrations of TTD. RT-PCR was used to detect the mRNA expression of mdr1 and NF-kappa B. Flow cytometry was used to assay the expression of P-glycoprotein (P-gp). Intracellular rhodamine 123 (Rho123) retention assay was applied to test the P-gp function.
RESULTSAfter treatment with 0.6 microg/ml doxorubicin for 24 hours, the expressions of mdr1 mRNA, NF-kappa B mRNA and P-gp in K562 cells were increased from 0.171 +/- 0.012, 0.783 +/- 0.090, 7.85 +/- 0.15 to 0.428 +/- 0.012, 1.075 +/- 0.047 and 73.68 +/- 1.84, respectively. The intracellular Rho123 retention was decreased from 711.9 +/- 63.6 to 347.8 +/- 60.6, indicating up-regulation of P-gp function (P<0.05). Pretreatment of K562 cells with 2.0 microg/ml TTD for 24 hours and then incubated for another 24 h with doxorubicin, the expressions of mdr1 mRNA, NF-kappa B mRNA, P-gp and up-regulation of P-gp function induced by doxorubicin were prevented in K562 cells (0.148 +/- 0.006, 0.627 +/- 0.098, 7.18 +/- 0.38 and 799.7 +/- 45.8, respectively P<0.05). But 0.5 microg/ml and 1.0 microg/ml TTD had little effect.
CONCLUSIONSTTD inhibits the expression of mdr1 mRNA, P-gp and up-regulated P-gp function induced by doxorubicin in a dose dependent manner. The mechanism of this effect may be down-regulation of NF-kappa B by TTD.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Benzylisoquinolines ; pharmacology ; Doxorubicin ; pharmacology ; Humans ; K562 Cells ; NF-kappa B ; metabolism ; RNA, Messenger ; genetics ; Up-Regulation ; drug effects
9.Calpain-I, calpastatin, caspase-3 and apoptosis in the human left atrium in rheumatic atrial fibrillation.
Yun-qing CHEN ; Lin WANG ; Xi SU ; Liang TAO ; Xu-fa CHEN
Chinese Journal of Cardiology 2006;34(4):303-307
OBJECTIVEThe aim of the present study was to detect the expression of calpain-I, calpastatin, caspase-3 and apoptosis in the left atria of patients with rheumatic heart disease (RHD), and to find the association of these factors. Also, it was intended to investigate the effect of the above factors on the mechanism of atrial fibrillation (AF).
METHODS43 patients with RHD undergoing valve-replacement were included, 15 patients with regular sinus rhythm (Group RSR), 8 patients with paroxysmal AF (Group AF1) and 20 patients with permanent AF (Group AF2). Western blot was used to examine the content of calpain-I, caspase-3 and calpastatin. The apoptosis index (AI) was measured by TUNEL.
RESULTS(1) Expression of calpain-I in group AF2 was increased to (344.0 +/- 101.9)%, and caspase-3 was increased to (394.0 +/- 99.4)% compared to group RSR (P < 0.01, respectively). Amount of calpastatin was reduced to (27.0 +/- 12.8)% (P < 0.01). The expressions of these proteins were unchanged in group AF1. (2) AI in group AF2 was higher than that in groups RSR and AF1 (P < 0.01). (3) In group AF2, the levels of calpain-I, caspase-3 and AI were positively relative to left atrial dimension and AF duration, P < 0.05 - 0.01, respectively, whereas calpastatin was negatively correlated with left atrial dimension and AF duration (P = 0.007 and P = 0.001, respectively). (4) The protein content of calpain-I was positively related with that of caspase-3 and AI (P < 0.01, respectively), and the content of calpastatin was negatively related with that of calpain-I and caspase-3 (P < 0.01, respectively).
CONCLUSIONSApoptosis of atrial cell increased in left atria and the protein contents of calpain-I, caspase-3 and calpastatin significantly altered during AF in humans with RHD. The observed interactions suggest that these factors compose a system to cause the structural remodeling and dysfunction of atria. The course may play a key role in promoting the onset and maintenance of AF.
Adult ; Apoptosis ; Atrial Fibrillation ; etiology ; metabolism ; Atrial Function ; Calcium-Binding Proteins ; metabolism ; Calpain ; metabolism ; Caspase 3 ; metabolism ; Female ; Heart Atria ; metabolism ; Humans ; Male ; Middle Aged ; Myocytes, Cardiac ; metabolism ; Rheumatic Heart Disease ; complications ; metabolism
10.Measurement Uncertainty Assessment for Determination of Allura Red in Puffed Food by HPLC
Ying CHEN ; Yun ZHAO ; Hua-Ming WANG ; Qing WANG ; Xi CHEN ; Dong-Qun XU ; Tao LI ; Feng YU ; Xiao-Yong WANG ; Jia-Fa LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(2):324-328
This study aimed to assess the measurement uncertainty of a new method for determination of allura redin food by high performance liquid chromatography (HPLC).The uncertainty of mathematical model of allura red is based on Europe for Analytical Chemistry(EURACHEM) guidelines.The sources and components of uncertainty were calculated,including recovery,working solution,sample mass,final volume,response of standard solution,response of sample solution.The expanded uncertainty was 0.0024 (k=2).Uncertainty of working solution was the most significant factor contributing to the total uncertainty,accounting for 86.2%.The uncertainty of volume accounted for the minimum at 0.025%.The developed method is simple and accurate,which can be used for the determination of allura redin puffed samples.