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Objective@#To observe the immunoreaction of offspring mice by ovalbumin (OVA) re-exposure after their mothers exposed to OVA during pregnancy.@*Method@#A prospective controlled study was conducted to observe mice after repeated OVA exposures at 6-8 weeks.Their mothers were exposed to OVA during different stages of pregnancy.The symptoms were recorded and scored.The levels of OVA-specific IgE in serum, interferon-γ(IFN-γ) and interleukin-4(IL-4) in supernatant of spleen primary lymphocytes in vitro were measured by ELISA.The results were analyzed by single factor analysis of variance or rank sum test.@*Result@#All the mice in each group had acute diarrhea.The diarrhea happened earliest (2 days) and most severe in the late pregnancy group (early pregnancy group 7.0±1.0; middle pregnancy group: 7.1±1.1; late pregnancy group: 9.9±2.2, P<0.01). The levels of absorbance of OVA-specific IgE in the pregnancy groups were higher than those of the control group.The absorbance of OVA-specific IgE in late pregnancy group was the highest (control: 0.27±0.06; early pregnancy group: 0.51±0.13; middle pregnancy group: 0.50±0.09; late pregnancy group: 0.63±0.13, P<0.01). There was no significant change in IFN-γ expression in cultured supernatant of spleen lymphocytes in each group (control: (133±7) pg/ml; early pregnancy group: (133±4) pg/ml; middle pregnancy group: (134±6) pg/ml; late pregnancy group: (132±4) pg/ml, all P value >0.05). The expression of IL-4 in the experimental groups was higher than that in the control group, especially in late pregnancy group(control: (25.3±2.4) pg/ml; early pregnancy group: (32.4±4.4) pg/ml; middle pregnancy group: (35.0±5.4) pg/ml; late pregnancy group: (47.1±5.8) pg/ml; P value all<0.01).@*Conclusion@#The allergic reaction of the OVA re-exposure progeny whose mothers were exposed to OVA in the late pregnancy period was most severe, suggesting that late pregnancy period might be the high risk stage of intrauterine sensitization, or"window period".
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Objective@#To study the tolerance to ovalbumin (OVA) in suckling mice whose mothers had different doses of docosahexenoic acid (DHA) microalgae oil (DMO) supplementation during pregnancy and lactation.@*Method@#According to different doses of DMO fed to mother mice during pregnancy and lactation, 66 suckling mice were divided into four groups. Suckling mice whose mothers were fed with 0.7% DMO were designated as low dose group (group L) (n=16), 2.1% DMO as middle dose group (group M) (n=16), 3.5% DMO as high dose group (group H) (n=17) and no DMO as control group (n=17). Before exposing to OVA, 8 suckling mice were killed in each group at 21-day-old. Remaining suckling mice were killed at 59-day-old after repeated OVA exposure. The serum polyunsaturated fatty acid (PUFA) levels of suckling mice were analyzed by high performance liquid chromatography (HPLC) at the age of 21- and 59-day.Histological examinations of jejunum were performed by HE staining and the mast cells in jejunum were observed by toluidine blue staining. OVA-IgE in serum, total IgA and OVA-IgA in the feces and IL-4 and IFN-γ in the supernatants of splenic mononuclear cells (SMC) were measured by ELISA. Real time PCR was performed to identify the gene expression of IL-10, TGF-β1 mRNA in SMC. Differences among groups were compared by one-way AVOVA and that between each group were compared by LSD.@*Result@#In group M and H, the serum levels of n-3DHA (108±29)μg/ml; (102±34)μg/ml vs.(40±19)μg/ml (F=12.052, P=0.000)and n-3 eicosapentaenoic acid (6.7±2.3)μg/ml; (7.7±2.0)μg/ml vs. (3.9±1.1)μg/ml(F=9.573, P=0.000) were significantly higher than that in control group at the age of 21-day. The serum levels of n-3DHA were higher in group H (17.1±2.9)μg/ml than that in control group (5.9±3.3) μg/ml after repeated OVA exposure at the age of 59-day (F=10.339, P<0.000). Compared with control group (53±12) pg/ml, the levels of IL-4 in SMC in group H (42±9)pg/ml were lower (F=2.484, P<0.05).@*Conclusion@#The serum levels of DHA in baby mice, whose mothers was fed with DMO during pregnancy and lactation, were significantly increased till adulthood. However, the effect on tolerance to OVA was limited.
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Children are special groups who grow rapidly,the characteristics of physical growth are discontinuous,and individual in different stages.Growth level,growth velocity and proportion of body need in consideration for a proper assessment and explanation of growth status in children.
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Objective: To investigate the islet ?-cell function and insulin sensitivity in adult male rats born intrauterine growth retardation caused by uterine placenta insufficiency. Methods: Bilateral uterine artery ligation (UAL) was performed on day 17 of gestation in the pregnant Wistar rats; sham-operated pregnant rats served as controls. The birth weights of the offspring in the UAL group were below the mean values of the birth weights of the control group more than 2SD, defined as IUGR. The tests were done in the adult male offspring (n=9 for each group). The glucose tolerance test was processed and Modified Beta-Cell function Index (MBCI) was calculated to evaluating the islet ?-cell endocrine function. Hyperinsulinemic- euglycemic clamp, the gold standard method to test the insulin resistance in the periphereal tissue was performed and insulin induced glucose infusion rate (GIR) was used to evaluate the insulin sensitivity. Results: (1) Though the mean birth weight of the newborn in the UAL group(4.49?0.56)g is much more than two standard deviations below the mean weight of that of the control group(6.16?0.30)g,P
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Objective To evaluate the intestinal barrier function of the patients by two-sugar absorption test with high performance liquid chromatography (HPLC). Methods Nineteen children with confirmed food hypersensitivity (FH) and 19 normal children were included in this study. The average age was (8.1?1.7) months. After 8-hour fasting, subjects drank test solution (5 g lactulose and 2 g mannitol per 100 ml) at the dose of 2 ml/kg. Five-hour urinary excretion ratio of lactulose/mannitol (L/M) were measured using high performance liquid chromatography (HPLC). The condition of HPLC was as follows: Sugar-PakI column with refractive index detector; Mobile phase: pure deionized water; Flow rate: 0.5 ml/min; temperature of column: 85 ℃. Results HPLC chromatogram of urine sample was stable. The retention time of mannitol and lactulose was 13.86 min and 9.27 min respectively. A significant rise in 5 h urinary L/M excretion ratios was found in children with FH (0.18?0.06) as compared to that of controls (0.05?0.03) (P
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Objective:To investigate the expressions of the insulin receptor(IR),insulin receptor substrate-1(IRS-1),insulin receptor substrate-2(IRS-2),phosphatidyl inositol 3-kinase(PI3K) and insulin-like growth factor-1(IGF-1) of the livers of the male adult rats born with intrauterine growth retardation(IUGR),and to find out the relationship between IUGR and insulin resistance in their adult life.Methods:The foods were available ad libitum throughout the pregnancy to the control group and the rats in the experimental group were fed 50% of the control group to build the IUGR animal model.Liver samples were collected when the male offspring grew up to 12 weeks old.The mRNA expressions of hepatic IR,IRS-1,IRS-2,PI3K,and IGF-1 were detected by RT-PCR.The protein expressions of hepatic IR,IRS-1,and IRS-2 were analyzed by immunohistochemistry.Results:The mRNA expressions of hepatic IR in the adult male rats with IUGR were significantly lower than control group[(0.41?0.06) vs(0.62?0.11),P0.05] in both groups.The mRNA and protein expressions of the hepatic IRS-1[mRNA:(0.77?0.20) vs(1.32?0.42),P0.05].The expressions of hepatic IGF-1 mRNA of the rats with IUGR were significantly lower than those in control group [(0.55?0.12) vs(1.22?0.34),P
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Objective:To investigate the effects of maternal malnutrition on the expression of the insulin signal transduction proteins in the liver of perinatal rats.Methods:(1)The dams were semi-starvation from the first day during pregnancy to build the animal model of intrauterine growth retardation(IUGR).On days 15,17,19,21(E15,E17,E19,E21)of gestation,the fetuses were delivered by cesarean section.The fetal rats,placenta,fetal liver and the fetal brain were weighed respectively.And the placenta weight to the body weight ratio(PWR),the liver weight to the body weight ratio(LWR),the brain weight to the body weight ratio(BWR)were calculated;The expressions of the hepatic insulin receptor(IR),insulin receptor substrate-1(IRS-1),insulin receptor subtrate-2(IRS-2),phosphatidyl inositol 3-kinse(PI3K)mRNA in the rats during perinatal period were detected by RT-PCR.Results:(1)The PWR of the IUGR fetus on E15 was lower than that in the control(P0.05).Conclusion:(1)In order to protect the development of brain,the fetus that suffered maternal malnutrition throughout the pregnancy had liver weight and body weight reduced much more than brain weight,which was known as "brain spare effects".(2)The expressions of hepatic IR mRNA and PI3K mRNA of the fetus were in normal ranges;But the expressions of the hepatic IRS-1 mRNA and IRS-2 mRNA of the IUGR fetus were decreased during the perinatal period.These may be related to the growth retardation and insulin resistance of the adult IUGR rats.
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Objective To clone and analyze the full-length cDNA of mouse integrin ?4, and repair the mutation sensible locei that caused the change of amino acids. Methods The cDNA of ?4 gene was amplified by RT-PCR using the total RNA extracted from mouse small intestine peyer’s patch. The PCR product was inserted into pMD19-T vector and then transformed into E. coli JM109. The positive recombinant clone was analyzed by restriction endonuclease and DNA sequencing. The mutation of ?4 cDNA that caused the change of amino acids was repaired. Results The cDNA of mouse ?4 had a length of 3 099 bp, and encoded a product of 1 032 amino acids. There were 12 bases pairs mutation of ?4 gene and the 6 base pairs causing the change of amino acids was repaired. Conclusion The cDNA of mouse ?4 is cloned successfully.
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The plasma free amino acids (PFAA) were measured in 37 healthy children (27 boys, 14 girls) under 4 years of age. Blood samples were drawn after an overnight fast. All the analyses were performed by elution chromatograph with a Beckman 121 MB Amino Acid Analyser. Data of plasma concentrations of 21 free amino acids and of 9 essential ones were collected. Among them, the concentrations of Alamine, Glutamine and Glutamate were on the top accounting about 40% of the total. In this study there revealed noither sex nor age differences in infants and children under 4 years of age, suggesting that in younger children the pool of amino acids is so expansible to fit their growth and development. The ratios of different amino acids were calculated. For example, NEAA/EAA was 1.5, Ala/Thr 2-3.0, Ala/Leu 2-2.5,Ala/ BCAA 0.6-0.7, Phen/Tyr 1 and Gly/Val 1. It seemed possible using these ratios to evaluate the status of malnutrition of young children
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Objective To develop HPLC method for determination of polyunsaturated fatty acid(PUFA) in breast milk.Method PUFA were extracted from breast milk with mixture of methanol: N-hexane:phosphoric acid(15:25:3).PUFA were derived by phenacyl bromide.The mobile phase was acetonitrile:water(81:19,v/v) at 1.0 ml/min with UV detection at 252 nm and column temperature was 35℃.The analytic column was Hypersil BDS C18(4.6mm?150mm,5 ?m).Results The average recovery was(98?5)% and the relative standard deviation of intra-day and inter-day were limited within 5.9%.Conclusion The method was stable,sensitive and accurate,and suitable for determination of PUFA in breast milk.