Cytogenetic test and clinical study on cryptic acute promyelocytic leukemia with ins (15; 17) / 中华血液学杂志

Objective@#To investigate the genetic screening methods for cryptic acute promyelocytic leukemia (APL) to further explore its clinical prognosis.@*Methods@#From June 2016 to November 2018, we collected 373 newly diagnosed APL cases. The patients were retrospected by the results of PML-RARα detections both by RT-PCR and i-FISH, those who harbored positive PML-RARα detection by RT-PCR and negative by i-FISH were chosen. Metaphase FISH and Sanger sequencing were further performed to verify these results.@*Results@#A total of 7 cryptic APL cases were discovered. These cases had tiny fragment of RARα inserted into PML in chromosome 15, formed ins (15;17) . The 7 cryptic APL cases had no PML-RARα gene subtype specificity, involving 5 cases in L subtype, 1 case in S subtype and 1 case in V subtype respectively. After the treatment of retinoic acid and arsenic or anthracyclines, 6 cases achieved complete remission, 1 case died of intracranial hemorrhage on the 6th day of therapy.@*Conclusion@#The size and covering position of PML-RARα probe should be taken into account when PML-RARα was performed by FISH on APL patients. Furthermore, combination with Metaphase FISH could improve the recognition of cryptic APL. There were no differences between the cryptic and common APL patients in terms of clinical features and treatment choices. Cryptic APL patients also had a good response to the therapy of retinoic acid and arsenic or anthracyclines.

Structural characteristics of the deciduous teeth of Tibetan miniature pigs / 南方医科大学学报

OBJECTIVE@#To explore the histological structure of the deciduous teeth and the tooth germs of Tibetan miniature pigs for studies of dental tissue diseases and tooth regeneration.@*METHODS@#The structure of the deciduous teeth of Tibetan miniature pigs was observed by X-ray. The ultrastructure of the enamel and dentin of deciduous teeth was characterized by scanning electron microscopy. The jaws and teeth were three-dimensionally reconstructed using Mimics software based on Micro-CT scanning of the deciduous teeth. Image J software was used to calculate the gray value and the mineralization density of the deciduous teeth. Hisotological structure of the tooth germ and the pulp tissue of Tibetan miniature pigs was observed using HE staining.@*RESULTS@#The deciduous teeth of Tibetan miniature pigs were composed of enamel, dentin and medullary pulp tissue. The permanent tooth germ were formed during the deciduous dentition. The enamel and dentin ultrastructure of deciduous teeth were consistent with that of human deciduous teeth. The enamel and dentin mineralization densities were 2.47±0.09 g/cm and 1.72±0.07 g/cm, respectively. The pathological structures of tooth germ and pulp tissue were similar to those of human teeth, and the pulp tissue of the deciduous teeth was in an undifferentiated state.@*CONCLUSIONS@#The deciduous teeth of Tibetan miniature pig have similar anatomy, ultrastructure and histopathological structure to human teeth and can serve as a good animal model for studying human dental tissue diseases and the mechanisms of tooth regeneration.

Significance of BIOMED-2 standardized IG/TCR gene rearrangement detection in paraffin-embedded section in lymphoma diagnosis / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the feasibility of detecting lymphoma with the application of BIOMED-2 standardized immunoglobulin/T cell receptor (IG/TCR) gene rearrangement system in formalin fixed paraffin-embedded (FFPE) tissue samples, and to discuss the relationship between the longest amplification fragment of extracted DNA and positive detection rate of different IGH V-J primers.</p><p><b>METHODS</b>DNA was extracted from 50 cases of FFPE tissue samples. Multiplex-PCR amplifications were performed and then the IG/TCR gene rearrangements were analyzed using BIOMED-2 standardized clonality analysis system.</p><p><b>RESULTS</b>(1)When the DNA concentration was diluted to 50-100 ng/μl from 100-500 ng/μl, the proportion of the longest amplification fragment (300-400 bp) of DNA was improved from 10.0% to 90.0% in 30 cases of diffuse large B cell lymphoma (DLBCL) wax roll samples (P<0.01). The positive rate of IGH+IGK was increased from 46.7% to 83.3%, the difference was statistically significant (P=0.006). The lengths of the longest amplification fragments of DNA were all longer than 300 bp in the paraffin section samples of DLBCL. The positive rate of IGH+IGK of these samples was 96.7%. The difference of the positive rate of IGH+IGK between the wax roll samples and the paraffin section samples had no statistical significance (P=0.195). (2)When the concentration of DNA was high, most of the longest amplification fragments of extracted DNA were 100 bp or 200 bp, and the detection rate of short fragment IGH FR3 was more stable than that of long fragment IGH FR1. (3)The clonality analysis of TCRG+TCRB in all 13 cases of peripheral T cell lymphoma samples showed positive results, while no positive IG/TCR clones were found in 7 cases of reactive lymphoid tissue hyperplasia in control group.</p><p><b>CONCLUSION</b>Dilution of DNA is the only method to improve not only the proportion of longest fragment amplification but also the detection rate of clonality. The detection rate of IGH FR3 would not be affected by the concentration of DNA. The application of BIOMED-2 standardized IG/TCR gene rearrangement system in FFPE tissue samples plays an important role in the lymphoma diagnosis.</p>

Application of three-probe fluorescence in situ hybridization panel in the diagnosis of pediatric B cell acute lymphoblastic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the three-probe fluorescence in situ hybridization (FISH) panel in the diagnosis of pediatric B cell acute lymphoblastic leukemia (B-ALL).</p><p><b>METHODS</b>Three-probe (TELAML1, BCR-ABL and MLL) FISH and conventional cytogenetic analysis were performed in 207 children with B-ALL.</p><p><b>RESULTS</b>In 207 B-ALL children, the three-probe FISH panel assay showed that 151 cases carried genetic aberrancies with a positive rate of 72.9%, including 44 cases with typical positive signal patterns and 148 cases with atypical signal patterns (among them 41 cases have multiprobe abberancy). The conventional cytogenetic analysis detected 53 cases chromosomal abnormality with a positive rate of 25.6%.</p><p><b>CONCLUSION</b>The detection rate of genetic abnormalities in newly- diagnosed pediatric B-ALL could be significantly improved by using three-probe FISH panel upon conventional cytogenetic analysis.</p>

Immunophenotypic, cytogenetic and clinical features of 113 acute lymphoblastic leukaemia patients in China

<p><b>INTRODUCTION</b>The analysis of immunophenotype of the leukaemic cells has been of great importance for the diagnosis, classification and prognosis of acute lymphoblastic leukaemia (ALL).</p><p><b>MATERIALS AND METHODS</b>One hundred and thirteen Chinese patients with ALL were immunophenotyped by fl ow cytometry and 74 cases were also subjected to karyotype analysis by G-banding technology.</p><p><b>RESULTS</b>Of the 113 Chinese ALL patients, 14.2% were identified as T-ALL and 85.8% as B-ALL. Myeloid antigen (MyAg) expression was documented in 34.9% of the cases analysed and CD13 was most commonly expressed MyAg in ALL patients (23.6%). MyAg positivity was higher in adult with ALL (47.6%) than in children with ALL (26.6%). Abnormal karyotypes were detected in 39 out of 74 (52.7%) cases. The clinical and biological characteristics of ALL patients between MyAg+ and MyAg- groups showed that increased white blood count (WBC) (>50 x 109/L), higher CD34 positivity and higher percentage of adult patients were found to be correlated with MyAg+ ALL.</p><p><b>CONCLUSION</b>Our results indicate that the immunophenotype did have relevance to the abnormal cytogenetic changes and clinical features in ALL. Flow cytometry immunophenotype has become the most important method for diagnosis and typing of ALL.</p>

Study of immunophenotype of acute lymphoblastic leukemia and its correlations to cytogenetics and clinical features / 中华检验医学杂志

objective To investigate the immunophenotype of Chinese patients with acute lymphoblastic leukemia(ALL)and its association to cytogenetics and clinical features.Methods In this study.a total of 113 Chinese patients with ALL were immunophenotyped by flow cytometry using a panel of monoclonal antibodies.50 000 cells were acquired and analyzed with Cell Quest and abnormal populations were recognized by CD_(45)/SSC gating.Among the 113 patients enrolled in this study,bone marrow cells of 74 patients were cultured for 72 hours to prepare for conventional chromosome detection.and karyotype was analyzed with R-banding technique.Results Of 113 ALL patients,14.2%(16/113)were identified as TALL,85.8%(97/113)as B-ALL Among the 106 out of 113 ALL cases,myeloid antigen(MyAg)expression was documented in 34.9%(37/106)and CD_(13) was the most commonly expressed MyAg in ALL patients(23.6%,25/106).MyAgs were more frequently associated with T-ALL(46.2%,6/13)than with B-ALL(33.3%,31/93)but there was no significant difieFence(X~2=0.825,P＞0.05)between the two groups.MyAg positivity in adult ALL patients(47.6%,20/42)was higher than that in children(26.6%,17/64).There was a significant difference between the two groups(X~2=4.948,P＜0.05).Abnormal karyotypes were detected in 39 out of 74(52.7%)patients.It showed that percentage of patients with high WBC count(＞50×10~9/L)in MyAg~+(48.6%,18/37)was higher than that in MyAg~-ALL types (20.3%,14/69).There was a significant difference between the two groups(X~2=9.191,P＜0.01).CD34 expression was found in 83.8%(31/37)of MyAg~+ and 53.6%(37/69)of MyAg~-cages.There was a significant difference between the two groups(X~2=9.527,P＜0.05).In addition,the percentage of adult ALL patients in MyAg~+ group(54.1%,20/37)was higher than that in MyAg~-group(31.9%,22/69).There was a significant difference between the two groups(X~2=4.948,P＜0.05).Conclusion lmmunophenotype analysis is useful for ALL diagnosis and classification and the immunophenotype has relevance to the abnormal cytogenetic changes and clinical features in ALL patients.