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We report a case of a pregnant woman with 46,XX karyotype and positive sex-determining region on the Y chromosome ( SRY) gene and her female fetus. Ultrasound examination at 12 +6 gestational weeks indicated a thickened fetal nuchal translucency, and 46, XX with a positive SRY gene was detected in the fetus through quantitative fluorescent-polymerase chain reaction and amniotic fluid karyotype. However, the ultrasound showed that the gender of the fetus was female, which was inconsistent with the phenotype of male syndrome with 46, XX combining positive SRY gene. The fluorescent in situ hybridization (FISH) revealed that the short arm of the Y chromosome translocated to the long arm of one of the X chromosomes, namely Yp11.3-Xq28. In addition, a copy number variation at Yp11.31p11.2 copy (about 1 MB) was found by chromosomal microarray analysis, which validated the result of FISH and was consistent with the mother. After genetic counseling, the parents chose to continue the pregnancy to full term, and no abnormalities were found in the infant during the follow-up.
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Objective To evaluate the necessity and feasibility by using two different PCR-based techniques for prenatal diagnosis of thalassemia. Methods 509 specimens for prenatal diagnosis of thalassemia were detected respectively by single tube multiplex PCR(STMP),reverse dot blot(RDB)or probe melting curve assay(PMCA)for commonα-thalassemia orβ-thalassemia mutations in double-blind tests. Samples with different detection results were confirmed with DNA sequencing analysis. Prenatal diagnosis of thalassemia results were verified or followed up after birth. Results In detectingα-thalassemia andβ-thalassemia,there was one case in STMP + RDB and another case PMCA indicating differentiating results. The detection sensitivity of STMP + RDB was higher than that of PMCA,and its difference can be used as an indication for maternal blood contamination. Conclusion The two PCR methods with different principles were necessary and feasible for the prenatal diagnosis of thalassemia. The two methods were complementary to each other ,which can ensure the reliability of the prenatal diagnosis results and reduce the defects of single technique. It is worthy to be popularized in clinical application.
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Objective:To explore the role of matrix metalloproteinase 12 (MMP12) in airway macrophages and pulmonary neu-rogenic substance P ( SP ) in the pathogenesis of asthma by analyzing their relationship in different categories of asthmatic patients.Methods:Twenty patients of asthma remission phase ( remission asthma group ) , twenty ones of mild acute exacerbation asthma (mild asthma group) and twenty healthy adults (normal control group) were included,respectively.After lung function was measured,the numbers of macrophage in induced sputum were counted.The expression levels of MMP12 mRNA and protein in sputum macrophages were detected by quantitative reverse transcription polymerase chain reaction and Western blot.The concentration of sputum SP was assayed by enzyme immunometric assay.Results: ( 1 ) Compared with the subjects in normal control group, forced expiratory volume in 1 second%predicted ( FEV1 ) and forced expiratory flow rates at 50% of the forced vital capacity % predicted ( FEF50 ) were much lower and the numbers of sputum macrophages were much higher in the patients in different asthmatic groups.Compared with the patients in remission asthma group,FEV1 and FEF50 were much lower in the ones in mild asthma group.(2) MMP12 expressions in the macrophages and the concentrations of SP in sputum were significantly increased in the patients in different asthmatic groups compared with those in normal control group;Furthermore,MMP12 and SP in mild asthma group were much higher than in remission asthma.(3) In all patients from different asthmatic groups,mRNA expressions of MMP12 in the macrophages were positively correlated with the levels of sputum SP or the numbers of sputum macrophages,whereas negative correlations between mRNA expressions of MMP 12 and FEV 1 or FEF50 were observed.Conclusion: The regulatory imbalance of macrophages′MMP12 and pulmonary neurogic SP may participate in the pathogenesis of asthma and become the potential targets for asthma therapy.
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Objective To investigate the clinical value of the probe melting curve analysis‐based PCR assay for the detection of three types of αT .Methods A total of 149 blood and prenatal archival DNA samples (6 of which were amniotic fluid samples)with three knownαT genes ,which included 63 carriers with Hb CS ,22 cases with Hb QS ,43 individuals with Hb WS and 1 double heter‐ozygote with Hb CS and Hb WS) as well as 20 samples with normalα‐globin gene sequence that had been confirmed by RBD com‐bined with DNA sequencing were selected to test the specificity of probe melting curve analysis by blind analysis .Results The probe melting curve analysis accurately detected 100 of the DNA samples previously characterized by S RBD combined with DNA sequencing .Conclusion Probe melting curve analysis‐based PCR assay for the detection ofαT is featured with rapidity and accuracy and can be applied to clinical and prenatal diagnosis .
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ObjectiveTo evaluate the reliability of the probe melting curve analysis (PMCA) based on real-time fluorescent PCR assay for the genetic diagnosis and prenatal diagnosis of β-thalassemia.MethodsA total of 135 cases of peripheral blood samples were collected from Zhuhai Municipal Maternity and Child Healthcare Hospital between 2008 and 2010.All the samples were performed preliminary diagnosis according to the hematological data.Of these,119 cases were diagnosed as β-thalassemia trait,4 cases were diagnosed as severe thalassemia and 12 cases were normal.In addition,44 cases of amniotic fluid and 8 cases of cord blood with high-risk for severe β-thalassemia were also collected.The diagnostic reliability of the PMCA assay and reverse dot blot assay was evaluated on 187 above-mentioned cases by direct DNA sequencing analysis in a double-blind study.ResultsThe genotypes of 185 cases were detected accurately based on the PMCA assay except for two cases:one heterozygote with Ini( ATG ＞ AGG) was omitted and another heterozygote couldn't be distinguished between CD43 ( G ＞ T) and CD37 ( G ＞ A ).For the RDB assay,only one heterozygote with CD71-72 ( + T) was not detected accurately in the above-mentioned cases.Compared with the DNA sequencing analysis,the sensitivity,specificity,negative predictive value,positive predictive value and diagnostic efficiency of the PMCA assay were 98.75％,100.00％,93.10％,100.00％ and 98.93％,respectively.The corresponding value of the RDB assay were 99.38％,100.00％,96.42％,100.00％ and 99.47％,respectively.There were no significant between-group differences in the diagnostic efficiency of the two assays ( P ＞ 0.05 ).The results of prenatal diagnosis were in complete concordance with the follow up results,after the birth or induced labour of the fetuses.ConclusionsThe PMCA assay can be used as an alternative and verified method of RDB assay for the genetic diagnosis and prenatal diagnosis of β-thalassemia.
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Objective To investigate the correlation between the erythrocyte indices and the genotypes of alpha thalassemia.Methods337 carriers with various genotypes of alpha-thalassaemia ( iron deficiency,alpha-thalassemia double heterozygote and homozygote,α-compounding β-thalassemia and abnormal hemoglobinopathy were excluded) were classified into three groups based on different genotypes of alpha-thalassaemia including silent thalassemia group (ST,83 cases),α-thalassemia trait group (TT,210cases) and intermediate thalassemia group( IT,44 cases),and 154 healthy adults were randomly choosed as normal control The erythrocyte indices involving in RBC,Hb,MCV,MCH,MCHC and RDW-CV were retrospectively analyzed and the difference of which was compared by analysis of variance and SNK test among aboved-mentioned groups.ResultsThere were statistical significance among groups about erythrocyte indices except Hb F.The order of the level of MCV and MCH was NC [( 86.6 ± 5.2) fl,( 29.5 ± 2.1 ) pg] ＞ST[(80.1 ±3.3) fl,(26.7±1.3) pg] ＞TT[(68.5 ±3.4) fl,(22.0 ±1.2) pg] ＞IT[(66.6±7.1)fl,(20.0 ±2.2) pg,F =580.67,761.19,P ＜0.05].And the size of RDW-CV was IT(22.3 ±3.4)％ ＞TT (14.9±1.2) ％ ＞ST(13.8±1.6)％ ＞NC(13.2±1.4)％(F=347.25,P＜0.05).In ST group,the value of MCHC of -α3.7/αα subgroup( 335.6 ± 8.0) g/L was higher than that of -α4.2/αα subgroup( 330 ±7.2) g/L and αTα/αα subgroup (328.4 ±9.5) g/L(F=6.07,P ＜0.05).Meanwhile,in IT group,the value of MCV of αTα/--SEA subgroup( 70.1 ± 7.2 ) fl was higher than that of -α3.7/--SEA subgroup ( 63.4 ±5.9) fl and -α4.2/--SEA subgroup ( 64.1 ± 4.0 ) fl ( F =5.55,P ＜ 0.05 ).However,the value of MCHC of αTα/--SEA subgroup( 289.7 ± 21.2 ) g/L was lower than that of other two subgroups [( 306.3 ± 8.4 ),(306.1 ± 8.7) g/L,F =8.72,P ＜0.05].Except Hb A2 and Hb F,there was positive correlation between the number of deleted α-globin gene and that of RBC and RDW-CV ( r =0.318 and 0.580,P ＜0.01 ).Nevertheless,there was negative correlation between the number of deleted α-globin gene and that of the other erythrocyte indices (r =-0.483,-0.827,-0.744 and -0.684,P all ＜0.01 ).ConclusionsThere is close correlation between the degree of anemia and the number of deleted α-globin gene characterized by Hb reduction and RBC increasing.In addition,the anemia degree of non-deletional Hb H disease is severer than that of deletional Hb H,which of Hb H disease with -α4.2/--SEA is severer than that with -α3.7/--SEA.
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Objective To study the proliferation inhibition effect by silencing PLCε gene expression with RNA interference in BIU-87 cells. Methods The specific short hairpin RNA recombinant plasmids were constructed by gene clone technology.The expression level of PLCε protein and mRNA were detected by Western blot and RT-PCR respectively after transfected recombinant plasmids into BIU-87 cells.The influence on proliferation was check by MTT.The changes of proliferating cell nuclear antigen(PCNA)were analyzed by immunocytochemical method,and the distribution of cell cycle was analyzed using flow cytometry. Results After transfected with the specific recombinant plasmids,PCNA expression was decreased 33.08%,and the analysis of cell cycle indicated that cells of G0/G1 phase were increased comparision with(40.75±2.30)%and(40.00±1.76)0A,and its G2/M phase cells(8.16±0.51)%were decreased strikingly compared with group control(31.20±1.76)%and group NP(35.94±1.58)%.Cells were blocked at G0/G1 phase,the cell proliferation was inhibited obviously. Conclusion PLCε may play an important role in proliferation of bladder cancer cells,which could be a potential target of biological treatment on bladder cancer in the future.