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Objective To explore the feasibility of fluorescence quantitative PCR(FQ-PCR)in prenatal diagnosis of the fetal RhD genotyping using free DNA from RhD-negative pregnant women.Methods The fetal RhD gene was amplified from 78 RhD-negative pregnant women with single fetus maternal plasma (gestation from 11 to 40 weeks).Rhe existence of fetal DNA was confirmed by amplification ofnine different polymorphic short tandem repeat loci(STR)and sex-determining region Y chromosome(SRY)gene.Exon5,7,10 and intron 4 were amplified by real-time polymerase chain reaction with TaqMan probe.The results of fetal RhD genotyping were evaluated retrospectively by the serologic analysis of infant cord blood.Results Among the 78 specimens,the SRY positive signals were detected from samples of 41 and were all identified male fetal through 8ex observation after newborn infants delivered from the women enrolled.The mean concentration of SRY gene reached(214.7±120.9)eopies/ml.RhD genotyping results of 70 cases were in complete concordance with the resets through serological detection of fetal cord blood after delivery.In addition,5 cases were false-positive.3 cages were considered inconclusive.The coincidence rate was 90%(70/78).From 5 false-positive cases,4 cases were identified as RhDel phenotype by detecting RHDl227A allele gene.The final accuracy rate of FQ-PCR was 95%(74/78)in the fetal RhD genotyping.Conclusion FQ-PCR analysis for noninvasive prenatal of fetal RhD genotyping could be useful in prevention and diagnosis of hemolytic disease of newborn.
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Objective: To study the effect of casein hydrolysate (CH) on hypertensive rats. Method: Hydrolysate of casein was desalinated by macroporous adsorption resins DA201C. Angiotensin converting enzyme (ACE) inhibitory activity and antihypertensive activity of casein hydrolysate at the dose of 500, 1 000 mg/kg were measured. Results: CH had significant inhibitory activity against ACE (the inhibitory rate 80.1%), and low concentration (500 mg/kg bw?d) had remarkable antihypertensive activity on spontaneously hypertensive rats (SHR), and extremely remarkable at high concentration (1 000 mg/kg bw?d). Conclusion: Casein hydrolysate showed antihypertensive activity in SHR.
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The respective sense and antisense oligodeoxynucleotides and their phosphorothiods, targeting at the bcr/abl, were the hallmark gene of Chronic Myelogenous Leukemia( CML) and c-myb correlates with CML. K562 cells, derived initially from a patient of CML blast crisis, were incubated with bcr/abl antisense oligodeoxynucleotides asODN or c-myb asODN or with both asODN in combinalion. All kinds of asODN could not only significantly inhibit K562 cells survival with the highest inhibitory rate of 64.7%?3.2%, but also dramatically inhibit DNA synthesis and the highest inhibitory rate was 85. 8 %?4.1%, decrease bcr/abl mRNA level almost completely and induce apoptosis significantly. The inhibition effect of antisense oligodeoxynucleotides phosphorothiodes s-asODN was stronger than that of unmodified asODN. The inhibition of the combination of bcr/abl and c-myb asODN was stronger than that of anyone alnoe. All the inhibitions exhibited in sequence-and time-dependent manner. We concluded that bcr/abl in pathogenesis of CML not only increased CML cells proliferation rate, but also decreased CML cells apoptosis rate. c-myb may participate in CML mainly by regulating bcr/abl. Trie results indicated that the modified asODN and multiple asODN targeting several oncogenes may advance to antisense gene therapy.
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Objective To explore the effects of lycopene on lifespan in drosophila and the underlying mechanism. Method Drosophila aged 30 d were reared in medium containing 0,2.5,7.5,22.5 ?g/g lycopene till they all die. The average lifespan and the average maximum lifespan were determined. Male drosophila aged 30 d were divided into control group and experiment group and cultured in medium supplemented with 0,22.5 ?g/g lycopene. The gene expression was determined 10,20 d later. Results The average maximum lifespan of male drosophila was increased with 2.5,7.5 ?g/g lycopene supp-lementation(P
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Objective: To study the effect of dietary nucleotides on DNA damage of thymocytes in mice. Methods: KM mice (n=30) , 5-6 weeks, were randomized into 3 groups,negative group (group 1), positive group (group 2) and nucleotides group (group 3). Mice in group 1 and 2 were fed nucleotide-free diet and group 3 nucleotide-supplemented diet (0.25% nucleotides). Mice in group 2, 3 were given single cyclophosphamide intraperitoneal injection of 150 mg/kg bw at day 21. DNA damage in thymus lymphocytes was evaluated 18 h after injection by single-cell gel electrophoresis (SCGE) assay. Thymus and spleen were weighed. Results: Dietary nucleotides had no effect on weights of thymus and spleen, but significantly decreased the number of damaged lymphocytes and the degree of damage. Conclusion: Dietary nucleotides may protect thymocytes DNA in mice from damage.
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Objective: To study the effects of exogenous nucleotides on immune function in mice.Methods: Sixty Kunming mice, (20?2) g, 5-6 weeks old were divided into three treatments groups (fed 0,0.05%,0.25% nucleotide-supplemented diet respectively). Each treatment was divided into two groups: normal group and immunodepression group. Mice in immunodepression groups were injected with cyclophosphamide (150 mg/kg bw) on the 2nd day in order to suppress the immune function and the mice in normal group were treated with normal saline at the same time. All mice were injected with SRBC on the 6th day. 0,0.05%,0.25% nucleotides were added into basic diet respectively and fed for 10 days. Body weight gain, organ index (thymus and spleen), SI, antibody anti SRBC and antibody secreting cell number in spleen were measured. Results: The body weight gain(P