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OBJECTIVE@#To explore the application value of next generation sequencing (NGS) in preimplantation genetic diagnosis of α/β complex thalassemia couple.@*METHODS@#The coding regions of α-globin genes (HBA1, HBA2) and β-globin gene (HBB) were selected as the target regions. The high-density and closely linked single nucleotide polymorphism (SNP) sites were selected as the genetic linkage markers in the upstream and downstream 2M regions of the gene. After NGS, the effective SNP sites were selected to construct the haplotype of the couple, and the risk chromosome of the mutation carried by the couple was determined. The NGS technology was used to sequence the variations of HBA1, HBA2 and HBB directly and construct haplotype linkage analysis for preimplantation genetic diagnosis.@*RESULTS@#Direct sequencing and haplotype linkage analysis of HBA1, HBA2 and HBB showed that two of the six blastocysts were α/β complex thalassemia, one was β-thalassemia heterozygote, two were α-thalassemias heterozygotes, and one was intermediate α-thalassemia. A well-developed embryo underwent preimplantation genetic diagnosis was implanted into the mother's uterus, and a healthy infant was born at term.@*CONCLUSION@#Preimplantation genetic diagnosis can be carried out by NGS technology in α/β complex thalassemia couples, and abortion caused by aneuploid embryo selection can be avoided.
Subject(s)
Female , Humans , Pregnancy , High-Throughput Nucleotide Sequencing , Mutation , Preimplantation Diagnosis , alpha-Thalassemia , beta-Globins/genetics , beta-Thalassemia/geneticsABSTRACT
Objective At present, the traditional model of urinary tract infection is not only complex in operation process but also easy to be infected, and the model with simple transurethral bacterial injection is not stable enough. This study established a rat model of foreign body-associated urinary tract infection. A spiral polyethylene tube (PT) was placed transurethrally into the bladder by transurethral inoculation with extended-spectrum β-lactamases-producing Escherichia coli (ESBLs E.coli).Methods SD female rats were randomly divided into blank control group, no foreign body groups at 7d and 14d, foreign body groups at 7d and 14d. ESBLs E.coli was injected transurethrally by foreign body into the bladder in foreign body groups. The same volumes of ESBLs E.coli and sterile saline were injected respectively in no foreign body and blank groups. Examination was made on bilateral renal gravity index, the numbers of white blood cells and neutrophils, the bacteriological and pathological changes in urine and kidney.Results The bilateral renal gravity index, the percentage of neutrophils and and the positive rates of renal and urine bacteria culture in foreign body groups at 7d and 14d were significantly higher than those in blank control group and no foreign body groups(P＜0.05). The number of white blood cells in foreign body group at 7d was significantly higher than those of blank control and no foreign body group(P＜0.05). The positive rates of renal and urine bacteria culture were 100% in the foreign body groups at 7d and 14d. The positive rate of urine bacteria culture in no foreign body at 7d was 33.3%. Under the light microscope, there were inflammatory changes in the kidney and bladder tissues of the rats in foreign body groups, and a few fibrous tissues were formed around glomeruli in foreign body group at 14d.Conclusion The establishment of a highly successful and stable rat model of urinary tract infection can be successfully achieved by injection of ESBLS E.coli through transurethral foreign body placed in bladder.
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<p><b>OBJECTIVE</b>To investigate the value of hemoglobin A(HbA) for screening thalassemia.</p><p><b>METHODS</b>A total of 2 000 adults' peripheral blood samples from Guangdong Women and Children Hospital from June 2013 to January 2014 were collected. The hemoglobin A(HbA) level was analyzed by the full automatic capillary electrophoresis technique, and the genotypes of thalassemia were detected.</p><p><b>RESULTS</b>The optimal cutoff values of HbAfor screening silent α-thalassemia, α-thalassemia trait, intermedia α-thalassemia and β-thalassemia trait were 2.85%, 2.65%, 2.25% and 3.45%, respectively; the areas under receiver operator characteristic (ROC) curve were 0.709, 0.839, 0.979 and 0.997 respectively; the sensitivities were 0.481, 0.721, 0.953 and 0.994, and the specificities were 0.846, 0.837, 0.929 and 0.969 respectively.</p><p><b>CONCLUSION</b>The optimal cutoff values of HbAfor screening different type of thalassemia based on our laboratory data are established by using ROC curve. According to the area under ROC curve, a satisfactory accuracy for screening intermedia α-thalassemia and β-thalassemia trait can be achieved by detecting hemoglobin Alevel.</p>
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<p><b>OBJECTIVE</b>To evaluate the detective efficacy of Chromogenic Coliform and Escherichia Coli Agar (CCEA).</p><p><b>METHODS</b>A new chromogenic medium CCEA prepared by Huankai laboratory was used to compare with a classical medium of violet red bile agar (VRBA), and other two Chromogenic media Agar I and Agar II by detecting separately 11 reference strains, thirteen sterile samples with Coliform or E.coli and other four samples, and the accordant rates of detection were observed.</p><p><b>RESULTS</b>CCEA had the good selectivity. To seven kinds of quality strains in the resultant analysis, CCEA with VRBA and Agar I had not shown salience difference (P > 0.05), and CCEA with Agar II had significant difference (P < 0.05). CCEA showed more advantages than the Agar II. To thirteen sterile samples with Coliform or E.coli in resultant analysis, CCEA with Agar I and Agar II had shown no significant difference (P > 0.05), while CCEA with VRBA had significant difference (P < 0.05). CCEA might be more advantageous than the VRBA. In analysis of the four actual samples of Coliform, CCEA with VRBA, Agar I and Agar II showed no significant difference (P > 0.05). The accordant rates were 90%, 71.88%, 86.25% and 81.25% respectively, showing CCEA > Agar I > Agar II > VRBA. To two actual samples of E.coli in the resultant analysis, the CCEA with Agar I and Agar II had not shown significant difference (P > 0.05). The accordant rates were 100% respectively.</p><p><b>CONCLUSIONS</b>The CCEA might be more advantageous than the VRBA, having the same efficacy as with Agar I and Agar II.</p>