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Objective:To evaluate the diagnostic capabilities of PCR-flow Fluorescence Hybridization technology in prenatal genetic diagnosis of thalassemia.Methods:8 005 cases of prenatal genetic diagnosis of thalassemia in Guangdong Women and Children Hospital from September 2017 to December 2020 were retrospectively analyzed. All samples were diagnosed by traditional genetic methods include multiple Gap-PCR, PCR-RDB, MLPA and Sanger sequencing. Meanwhile, PCR-flow Fluorescence Hybridization technology was used as a verification platform for detecting common mutation sites of thalassemia. The results were analyzed to evaluate the diagnostic capabilities of PCR-flow Fluorescence Hybridization technology compared with traditional methods in prenatal genetic diagnosis of thalassemia.Results:By traditional methods, 1 939 cases (24.22%, 1 939/8 005) were normal and 6 066 cases (75.78%, 6 066/8 005) were diagnosed as thalassemia, including 4 513 cases of α-thalassemia, 1 475 cases of β-thalassemia, and 78 cases of αβ-thalassemia. By PCR-flow Fluorescence Hybridization technology, 7 845 samples were successfully diagnosed after initial interpretation by software. Compared with traditional methods, all the sensitivity, specificity and accuracy were 100%. The other 160 samples which failed in the initial interpretation can be successfully interpreted after review or manual interpretation.Conclusion:There were no differences between the two methods on the detecting of common mutation sites of thalassemia.
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Objective@#To explore the application of chromosomal microarray analysis (CMA) in neonatal birth defects and further determine the frequency of chromosome imbalances in neonates with birth defects.@*Methods@#Retrospective analysis was performed in 121 neonates with specific features, such as distinctive facial features, congenital heart disease, congenital malformation, neonatal decreased responsive, hypotonia, seizures and others at the Department of Neonatology, Guangdong Women and Children Hospital from May 2016 to November 2017.All the cases were analyzed by using chromosomal microarray analysis (CMA).@*Results@#A total of 23 (19.0%) patients were identified with pathogenic CNVs, 3 patients (2.5%) with uncertain clinical significant CNVs.There were 5 patients (4.1%)with chromosome numerical abnormalities, 12 patients (9.9%)with microdeletion/microduplication syndrome, 6 patients (5.0%) with chromosome deletion or duplication.All groups whose incidence was sorted from high to low were facial characters(30.3%), congenital malformation(21.6%), neonatal decreased responsive(9.1%) and other indications(6.7%).@*Conclusions@#CMA is a valuable clinical diagnostic tool that allows precise identification of chromosome imbalances as the cause of birth defects in neonates.CMA can detect many more clinically relevant genomic abnormalities than conventional cytogenetic study and assist the clinician in diagnosis, early neonatal intervention, and genetic counseling.
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<p><b>OBJECTIVE</b>Diagnosis and prenatal diagnosis to a family of hemoglobin variant with α-thalassemia.</p><p><b>METHODS</b>Whole blood cell analysis, hemoglobin analysis by capillary zone electrophoresis (CZE), Gap-PCR, polymerase chain reaction-reverse dot blot (PCR-RDB) assay and DNA sequencing.</p><p><b>RESULTS</b>Hb Zurich Albisrieden with α°-thalassemia lead to severe anemia. The genotype of fetus is also Hb Zurich Albisrieden with α°-thalassemia.</p><p><b>CONCLUSION</b>Abnormal hemoglobin with α-thalassemia may lead to severe anemia, Prenatal diagnosis of thalassemia has the vital significance for eugenic birth.</p>
Subject(s)
Adult , Child, Preschool , Female , Humans , Male , Pregnancy , Young Adult , Base Sequence , Fetal Diseases , Blood , Diagnosis , Genetics , Hemoglobins, Abnormal , Genetics , Metabolism , Molecular Sequence Data , Prenatal Diagnosis , alpha-Thalassemia , Blood , Diagnosis , Embryology , GeneticsABSTRACT
Objective To devolope a method for extracting DNA from dried blood spots (DBS)and optimizing the operating procedure,which could be applied to clinical gene diagnosis of thalassemia.And the cross contamination of DBS punching and the storage stability of DBS were studied.Methods A total of 1 50 blood specimens were collected,and DBS were prepared.Circles (3 mm in diameter)were punched in the DBS,and eluted with lysis buffer.The eluting method and operating procedure were opti-mized.Genomic DNA extracted from the elution solution by magnetic beads,and were performed thalassemia gene test.Finally jud-ging whether the results of DBS and whole blood were consistent.Two methods of thalassemia gene test were used in DBS and the compatibility of DBS processing method was verified.Judging whether there was cross contamination of DBS punching by the thalassemia gene test results of blank hole which were punched in the blank filter paper between thalassemia positive DBS.The DBS storage stability in thalassemia gene test was verified by detecting the DBS which were dry stored at room temperature for 6 and 9 months.Results 5 circles (3 mm in diameter)DBS were vibrating eluted at 55 ℃ for 1 hour,the DNA concentration extracted from the elution solution was 10-20 ng/μL,which was dissolved in 50 μL solution,and the DNA quality was good.The thalassemia gene test results of DBS and whole blood were the same,and the DBS results of two thalassemia gene test methods were the same too. The cross contamination of DBS punching was not detected in thalassemia gene test.The DBS which were dry stored at room tem-perature for 6 and 9 months could be stably performed thalassemia gene test.Conclusion DBS could be used to perform thalassemia gene test,which is accurate,convenient and stable.It is an ideal way for specimen referral of thalassemia gene test.
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Objective To retrospectively analyse the efficacy and the influencing factors of plasma exchange(PE)in patients with chronic severe hepatitis.Methods Sixty patients of chronic severe hepatitis were included.Results The effective rate was 61.7%.The efficacy of PE was closely related to age,clinical stage,pre-treatment serum bilirubin values,prothrombin activity and incidence of complications(P0.05).Conclusion PE can significantly improve the survival of patients with chronic severe hepatitis.Especially for the patients under 60 years old,those with low serum TBIL level and high PTA,and those in the early clinical stage,PE treatment would achieve a better efficacy.But the improvement of prognosis of severe hepatitis still depends on the early specific treatment and active prevention of complications.
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AIM To study the cancer promoting effects of N nitrosodiethylamine (DEN) and 2 Acetylaminofluorene (2 AAF). METHODS Medium Term Rat Liver Bioassay (MTRLB). Male SD rats were initially given a single dose (200 mg?kg -1 ) of DEN ip and starting 2 weeks later, were treated with 10, 33 and 100 ppm DEN in drinking water, or with 2 2, 6 6 and 22 mg?kg -1 2 AAF by gavage for 6 weeks. All rats were subjected to two thirds partial hepatectomy at week 3 and killed at the end of week 8. Carcinogenic potential was scored by comparing the numbers and areas in induced glutathione S transferase placental form (GST P) positive foci in the liver with those of corresponding control group given DEN alone. RESULTS Both DEN and 2 AAF caused the increases of the numbers and areas of GST P positive foci in the liver, and showed dose response relationship. CONCLUSION Both DEN and 2 AAF shows cancer promoting effects, and MTRLB was a convenient, economical and effective tool to study the cancer promoting effects of test chemicals.
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The expression, amplification and rearrangement of c-myc and c-erbB-2 genes in thyroid tumor were studied. 32P-dATP-labelled probes of c-myc, c-erbB-2 and v-sis DNA fragments were used to hybridise with the cellular total RNA. We found that c-myc and c-erbB-2 oncogenes were expressed in all samples. The levels of c-myc RNA were three-to eleven-fold higher in 9 out of 15 cancer samples when compared with that in normal tissues. In 7 of 15 cancer samples, c-erbB-2 gene was overexpressed 10~50 times higher than normal. No v-sis RNA was detected in all the samples. Southern blot hybridisation showed that rearrangement of c-myc oncogene were observed in 4 cancer samples, of which one showed a 150-fold amplification of c-myc gene. No amplification or rearrangement of c-erbB-2 gene was detected. These data indicate that the activat.:on of c-myc and c-erbB-2 oncogenes may contribute to the development and/or maintenance of the malignant phenotype of the thyroid carcinomas.
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Objective To study the relationship between the ultrastructural changes of corneal matrix collagen fibrils diameter and the postmortem interval.Methods Twenty-eight rabbits were killed by the method of air embolism and the samples of cornea were separately cut down in 0,6,12,18,24,48 and 72h after death.The ultrastructure of the collagen diameter was observed by transmission electro microscopy(TEM)and the 4 parameters of area(Y1),perimeter(Y2),average-diameter(Y3)and mean-diameter(Y4)were measured by a computer image technique and analyzed by Excel 2000 and SPSS 10.0 software.Results The 4 parameters of Y1,Y2,Y3 and Y4 were respectively increased from 1 131?53nm to 1 628?26nm,from 132.8?23nm to167.5nm,from 38nm to 45nm and from 37.71?6nm to 44.89?5nm within 72 hours after the death.Conclusion The corneal matrix collagen diameter increase regularly with the postmortem interval in 72h after death,and the parameters of Y1,Y2,Y3 and Y4 could be used as a new reference mark for timing the postmortem interval after death.