Karyotyping analysis of 396 newborns with congenital malformations and chromosomal abnormalities and the associated phenotypes / 中华实用儿科临床杂志

Objective To reveal the chromosome abnormalities and their relationship with the clinical phenotype of neonates with congenital malformation.Methods Karyotype analysis of peripheral blood lymphocytes was performed on 396 newborns with congenital malformation,who were recruited at the Children's Hospital Affiliated to Soochow University from Jan.2006 to May 2012,chromosome karyotypes were prepared with neonatal peripheral lymphocytes by conventional G-banding technique.Results 1.Of 396 newborns,159 (40.2％) cases were detected to have chromosomal abnormalities,including karyotype first reported domestically and internationally in 3 cases.2.Trisomy-21 (Down's syndrome),which was the most common abnormal karyotype,was seen in 130 cases,accounting for 81.8％,of whom 119 cases show the standard type,10 cases accompanied by the Robertsonian translocation involving group D or group G,and 1 case accompanied by sexual chromosomal abnormality:inv(Y) (p1 1 q 1 1),+ 21.3.Other common karyotype abnormalities were as follows:del (5) (p 1 2-14) (cats cry syndrome) in 4 cases,trisomy-18 (Edwards syndrome)in 4 cases,45,XO (Turner' s syndrome) in 4 cases,inv (9) (p1 1 q1 2-21) in 4 cases,trisomy-X (super female syndrome) in 1 case,rob(13;14) in 1 case,trisomy-8 in 1 case and del(18) (q22) in 1 case.4.Special faces were seen in 147 cases (92.5 ％),congenital heart disease in 97 cases (61.0％),low birth weight in 72 cases (45.3 ％),congenital anal atresia in 13 cases(8.1％),multiple malformations in 11 cases (6.8％),intestinal malformations in 10 cases (6.2％),extrinsic genital abnormalities in 9 cases(5.7％),meow-like cry in 4 cases(2.5％),limb edema in 4 cases (2.5％),fingers and toe abnormalities in 6 cases(3.6％),congenital brain dysplasia in 6 cases (3.6％),webbed neck in 5 cases(3.1％) and cleft lip and palate in 3 cases(1.8％).Conclusions Chromosomal abnormality is an important factor leading to neonatal birth defects,of which special face,congenital heart disease,low birth weight,and multiple malformations are the main clinical manifestations of chromosomal diseases.

Up-regulation of TIMP-2 expression promotes SHI-1 leukemic cells proliferation and infiltration in immunodeficiency mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>MMPs and TIMPs play important roles in tumor angiogenesis and invasion. Studies have shown that TIMP-2 has two roles in tumor invasion. However, its role in leukemic infiltration has not been well investigated. This study explored the roles of TIMP-2 in extramedullary infiltration of acute monocytic leukemic SHI-1 cells both in vitro and in vitro.</p><p><b>METHODS</b>A retroviral vector carrying the human TIMP-2 cDNA was constructed and transfected into the monocytic leukemic cell line SHI-1. The expression of TIMP-2 in the positive clones was determined. The proliferation of SHI-1 cells was examined by MTT assay. Trans-Matrigel invasion assays were used to investigate the infiltration ability in vitro. SHI-1 cells were intravenously injected into pre-treated nu/nu mice to investigate the infiltration ability feature in vitro.</p><p><b>RESULTS</b>The expression of TIMP-2 on the cell membrane was significantly elevated in SHI-1/TIMP-2 cells. Over-expression of TIMP-2 promoted the cells proliferation and the invasions in vitro. The SHI-1/TIMP-2 cells demonstrated higher infiltration ability when intravenously injected into nu/nu mice.</p><p><b>CONCLUSION</b>Over-expression of TIMP-2, especially on the cell membrane, may play important roles in promoting the proliferation and infiltration of SHI-1 leukemic cells.</p>

Clinical and genetics characteristics of patients with monosomal karyotype acute myeloid leukemia patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical and genetics characteristics of patients with monosomal karyotype acute myeloid leukemia (MK-AML).</p><p><b>METHODS</b>The karyotypes of 3743 patients with newly-diagnosed de novo AML were analyzed, which had identified 153 cases with MK-AML, for whom the clinical and genetics characteristics were analyzed.</p><p><b>RESULTS</b>There were 2056 patients (54.9%) among all patients. A total of 153 patients fulfilling the criteria for MK-AML were identified, which comprised 93 males and 60 females, with a median age of 54. The median white blood cell count on presentation was 4.4×10 (9)/L. One hundred and forty-five cases (94.8%) have fulfilled the criteria for complex karyotype (≥ 3 chromosomal abnormalities). Although the monosomy could be found with all autosomes, chromosome 7 has been most frequently involved (38.56%, 59/153).</p><p><b>CONCLUSION</b>MK-AML is a distinct cytogenetic subtype of AML. Monosomy 7 is frequently detected among MK-AML patients. The monosomal karyotype is common among elder patients with AML.</p>

A clinical and laboratory study of chronic myeloid leukemia with atypical BCR-ABL fusion gene subtypes / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical and laboratory features of chronic myeloid leukemia (CML) with atypical e14a3 and e19a2 BCR-ABL fusion gene subtypes.</p><p><b>METHODS</b>We retrospectively analyzed a cohort of CML patients with Ph chromosome positive confirmed by cytogenetic and FISH but classical e13a3(b2a2), e14a2(b3a2)and e1a2 fusion transcripts negative identified by conventional real-time quantification RT-PCR (RQ-PCR). Further RQ-PCR was done with the forward primer and reverse primer designed to detect rare atypical BCR-ABL fusion genes including e14a3 and e19a2 transcripts. Direct sequencing analysis was performed on the PCR products and mutations in the BCR-ABL kinase domain were detected. The clinical data of patients were retrospectively analyzed.</p><p><b>RESULTS</b>Six CML patients were found to carry t(9;22) abnormality and BCR-ABL rearrangement confirmed by FISH but classical BCR-ABL fusion genes negative detected by RQ-PCR. Further RQ-PCR and sequencing analysis confirmed the fusion of BCR exon 14 and ABL exon 3 in five CML patients (case 1-5) and the fusion of BCR exon 19 and ABL exon 2 in one CML patient (case 6). E255K and I293T IM-resistant mutations were detected in case 1 and 2, respectively. Among five cases with e14a3 transcripts, four were CML-CP, one CML-AP. Four patients were male and one was female. The median age was 48 years. The patient (case 6) with e19a2 transcripts was 40-year-old female with a diagnosis of CML-CP and PLT count was more than 1 000×10⁹/L. Imatinib (IM) therapy was administer in case 1, 2, 3, 4 and hematopoietic stem cell transplantation (HSCT) was undergone in case 5 after hydroxyurea (Hu) or interferon failure. Case 1 who had E255K IM resistant mutation, responded poorly to IM but obtained a complete cytogenetic remission (CCyR) after a substitution of dasatinib for IM. Case 2 and 3 achieved CCyR 6 months later after IM treatment and had been maintained well with IM despite I293T mutation in case 2. Case 4 attained CCyR 3 months later after IM treatment but relapsed and died soon. Case 5 was still in CCyR after HSCT. Case 6 with e19a2 transcripts got complete hematologic response after Hu treatment and CCyR was achieved soon after IM therapy.</p><p><b>CONCLUSION</b>Incidence of CML with atypical transcripts is extremely low. They could benefit from tyrosine kinase inhibitors or HSCT. Rare and atypical BCR- ABL fusion gene subtypes could be missed by conventional RQ-PCR.</p>

Genetics analysis of two childhood acute myeloid leukemia patients with variant t(8;21) / 白血病·淋巴瘤

Objective To report two childhood acute myeloid leukemia (AML) patients with t(8;20)(q22;q13) and t(1;8;21)(q32;q22;q22) respectively,as variant t(8；21).Methods Chromosome preparation of bone marrow cells were made using short-term culture and karyotypic analysis was carried out using R and G-banding techniques.Interphase-fluorescence in situ hybridization (I-FISH) and metaphase-FISH (M-FISH) were performed using dual color,dual fusion AML1-ETO probe to detect the AML1-ETO fusion gene.Multiplex RT-PCR was used to demonstrate the expression of AML1-ETO fusion transcript.Results The karyotype of bone marrow cells for these two childhood AML patients were 45,X,-Y,t(8;20)(q22;q13)[12]/46,XY[3](case 1) and 46,XX,t(1;8;21)(q32；q22;q22)[18]/46,XX[2](case 2),respectively.I-FISH and M-FISH confirmed that they all had the AML1-ETO fusion gene and variant t(8;21).The AML1-ETO fusion transcript in both patients was detected by RT-PCR.Conclusion t (8;20)(q22;q13) and t (1;8;21)(q32;q22;q22) are variant t (8;21) in nature.It is important to combine the conventional karyotypic analysis with D-FISH and multiplex RT-PCR to determine the nature and prognosis of AML patients with variant t(8;21).

One case of acute promyelocytic leukemia with 3'RARα submicroscopic deletion / 白血病·淋巴瘤

Objective To report a rare case of M3r subtype of acute promyelocytic leukemia (APL)with 3'-end of RARα (3'RARα) submicroscopic deletion, and the characters of morphologic, cytogenetic,molecular genetic and molecular biology studies. Methods Chromosomes of bone marrow (BM) cells were prepared with direct method and short-term culture method, and R-banding technique was used for karyotypic analysis. Fluorescence in situ hybridization (FISH) assays were performed on fixed BM cells using the following specific DNA probes: CEP X/Y alpha satellite DNA probe, LSI PML-RARα dual-color dual-fusion and LSI RARα dual-color break apart probes. A quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) was performed to detect the PML-RARα transcript. A multiplex nested RT-PCR was also performed, which may simultaneously detect the fusion genes derived from 29 chromosomal aberrations in acute leukemia including PML-RARα, PLZF-RARα and NPM-RARα fusion transcripts. Results R-banding analysis revealed a karyotype of 45,X,-Y[6]/46,XY[8], FISH using CEP X/Y probe further confirmed Y-chromosome loss. FISH analysis with RARα dual-color break apart probe demonstrated a deletion of the entire 3'-end of one allele of RARα gene. Cytogenetic, FISH and RT-PCR analyses showed no PML-RARα,PLZF-RARα, NPM-RARα, NuMA-RARα and STAT5b-RARα rearrangements. Conclusion A new RARαrearrangement involving 3'RARα submicroscopic deletion in APL without X-RARα fusion has been identified.FISH analysis with RARα dual-color break apart probe is a useful method for characterization of this abnormality, but its molecular consequences remain to be elucidated.

The chromosomal aberration detected by comparative genomic hybridization in lung cancer / 中国医师杂志

Objective To understand the molecular aberration at whole genomic level,CGH (comparative genomic hybridization) was used to investigate genetic abnormality in lung cancer.Methods Comparative genomic hybridization was performed in 17 cases to detect the global genomic aberration in cancer tissue cells.Results All of 17 cases detected by CGH showed chromosomal aberrations.The average numbers of chromosomal gains and losses in each case were 7.0 and 4.8 in NSCLC and 8.4 and 9.6 in SCLC,respectively.The frequency of gains and losses on chromosome had no significant differences between NSCLC and SCLC.The frequencies of gains on chromosomal arms 3q24 -28 and 11q13(58.3％ and 58.3％ ) in NSCLC were significantly higher than that in SCLC(0％ and 0％ ) ( P ＜0.05 and ＜0.05,respectively).Conclusions The cytogenetic aberration generally existed in lung cancer cells.Several regions ( more than one) of chromosomal aberration were involved in the carcinogenesis of NSCLC and SCLC.The regions and frequencies of chromosomal aberration in NSCLC were somewhat different from that in SCLC,which might result in the different biological behavior of the two types of lung cancer.The chromosomal aberration might be served as a marker to differentiate the two types of lung cancer.

A clinical and laboratory study on acute myeloid leukemia with t(6;9)(p23;q34) / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the clinical and laboratory features of 6 cases of acute myeloid leukemia (AML) with t(6;9)(p23;q34).</p><p><b>METHODS</b>Chromosome preparation of bone marrow cells was performed with regular method. R-banding by heating using Giemsa banding technique (RHG) was used for karyotype analysis. The immunoprofile was studied by flow cytometry (FCM) using a panel of monoclonal antibodies. Chromosome painting was performed by using whole chromosome paint probes for chromosomes 6 and 9 in all the 6 cases. The expression of fusion gene DEK/CAN and FLT3-ITD mutation were analyzed by reverse transcription-PCR(RT-PCR).</p><p><b>RESULTS</b>The t(6;9)(p23;q34) was found in all the 6 cases including 4 cases of M2 and 2 cases of M4. Blast cells were positive for CD13 and CD33 in 6 patients, for HLA-DR in 4 patients, for CD34 and CD117 in 3 cases, for CD38 or CD15 each in 1 case, respectively. A reciprocal translocation between chromosome 6 and 9 was confirmed by chromosome painting technique in the 6 cases. The DEK/CAN fusion gene was found in all the 6 cases, FLT3-ITD mutation was detected in three of them. Follow-up showed that 3 patients died with a survival time of 3 months, 5 months and 6 months, respectively. The other three obtained complete remission and are still alive.</p><p><b>CONCLUSION</b>The t(6;9)(p23;q34) is a rare recurrent abnormity. AML with t(6;9)(p23;q34) has unique clinical and laboratory features and its prognosis is poor in most cases.</p>

Chromosome study on chronic lymphocytic leukemia using CpG-oligodeoxynucleotide as immunostimulant agent / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate whether CpG-oligodeoxynucleotide (CpG-ODN) can improve the detection rate of the karyotypic abnormalities in chronic lymphocytic leukemia (CLL).</p><p><b>METHODS</b>The bone marrow (BM) or peripheral blood (PB) cells from 57 cases of CLL were collected and cultured with CpG-ODN DSP30+interleukin-2 (IL-2), phytohemagglutinin (PHA), pokeweed (PWM) or IL-2, respectively. Five days later cells were harvested for chromosome preparation. Karyotypic analysis was done using R banding technique. Panel fluorescence in situ hybridization (FISH) was carried out on 19 cases of CLL with normal karyotypes using the following probes: Cen12, D13S25, Rb1, ATM, p53, MYB and IgH. Genomic DNA from 21 cases of them was extracted from BM or PB leukocytes. The immunoglobulin variable heavy chain (IgVH) was amplified by polymerase chain reaction (PCR) and sequenced. CD38 and ZAP70 expressions in the leukemic cells were determined by flow cytometry (FCM).</p><p><b>RESULTS</b>The detection rate of karyotypic abnormalities in the CpG-ODN+IL-2 group (43.85%) was obviously higher than that in the PHA (15.09%), PWM (17.31%) and IL-2 (3.13%) groups (P<0.01). Fifty-two types of karyotypic abnormalities were found. Among them, trisomy12 (+12) or +12 with other abnormalities were the most common, while translocations were the most frequent structural abnormalities including 3 unbalanced and 11 balanced translocations, among them 7 had rearrangements involving 14q32. Thirteen cases showed one or more abnormalities on FISH including trisomy 12 and p53 deletion each in one case, IgH rearrangement and partial deletion each in one case, 13q14.3 deletion in 11 cases of which 5 cases also had Rb1 deletion, 1 case had Rb1 partial deletion. No case with ATM or MYB deletions was found. PCR detected IgVH mutations in 10/21 cases. FCM showed 10/45 cases were CD38 positive, but 35 /45 were CD38 negative, 11/27 cases expressed ZAP70, but 16/27 did not. Among the 26 cases examined for CD38 and ZAP70 expressions simultaneously, 5 cases were CD38+ZAP70+, 13 were CD38-ZAP70-, 6 were CD38-ZAP70+, and 2 were CD38+ZAP70-, respectively. Statistic analysis showed a correlation between complex karyotype and IgVH without mutation, but no association between karyotype and CD38 or ZAP70 expression was observed.</p><p><b>CONCLUSION</b>CpG-ODN immunostimulation can obviously raise the detection rate of abnormal karyotypes, especially translocations in CLL. FISH is an important complement to conventional karyotypic analysis. The combination of both methods can provide more comprehensive genetic information for CLL.</p>

A clinical and laboratory investigation of acute promyelocytic leukemia with tetraploid clone characterized by two t(15 ;17) / 中华检验医学杂志

Objective To investigate clinical and experimental features of APL with tetraploid clone characterized by double t (15 ; 17). MethodsFive cases of APL with tetraploid clone characterized by double t(15;17) were chosen. Cytogenetic examination of bone marrow was performed with bone marrow or short-period culture. R banding technique was used for karyotype analysis. DNA content in one case was determined by flow cytometry. Immunophenotyping was performed by using a panel of monoclonal antibodies :CD2, CD13, CD15, CD33 and CD34. PML/RARα fusion gene was detected by interphase FISH using dualcolor PML/RARα probe in one case and by RT-PCR in two cases. ResultsAll cases were male with a median age of 38. Their marrow cell morphology examination showed marked hyperplasia with large leukemic cells that had bizarre nuclear configuration. Chromosome analysis revealed that a leukemia clone with tetraploid or near-tetraploid karyotype characterized by double t(15;17) (q22 ;q12) in five cases, of which, one also had a diploid clone with t(15;17) and a normal cell;two had some cells with normal karyotypes.PML/RARα fusion gene was detected by FISH in one of 5 cases and by RT-PCR in 2 of 5 cases. CD33 expression was found in one case. CD13 and CD33 expressions were seen in the other four cases, of which,CD34 or CD2 co-expression was found in one case and in two cases respectively. The result of DNA content showed a single peak which indicated only tetraploid clone whose DNA index was 1. 998 with CV of 8. 2%.All patients obtained complete remission after the treatment with ATRA and/or arsenic trioxide. Conclusions These results indicate that API, patients with tetraploidy and near-tetraploidy have giant and bizarre blasts.Most patients have short-type PML/RARα transcripts. The tetraploidy in APL does not appear to affect the response to treatment of ATRA.

The quantitative assay and clinical significance of JAK2V617F mutation in 131 patients with chronic myeloproliferative disorders / 中华内科杂志

Objective To investigate the frequency and mutational status of JAK2V617F mutation in Chinese patients with chronic myeloproliferative disorders(CMPD) and to study the relative quantification of mutated JAK2 mRNA and the clinical significance. Methods JAK2V617F mutation and the mutational status were screened with amplification-refractory mutation system polymerase chain reaction(ARMS-PCR), the relative quantification of mutated JAK2 mRNA was studied by using capillary electrophoresis. Results A higher prevalence of JAK2V617F in either the heterozygotc or homozyote status in essential thrombocythemia (ET) was observed in elderly patients with ET (P<0.05). The presence of JAK2V617F was found to be significantly correlated with the age at diagnosis (P<0.05); patients with age ≥ 60 years showed significantly higher JAK2 mutated RNA levels than those with age < 60 years (P<0.05); the presence of JAK2V617F in polycythemia vera (PV) and ET was found to be significantly associated with higher hemoglobin level and higher leukocyte count (P< 0.05). In addition, higher leukocyte count was observed in homozygous ET patients than in heterozygous ET patients (P<0.05). The frequency of JAK2V617F mutation and the prevalence of homozygote in PV patients were higher than those in ET patients (P<0.05). The differences of JAK2V617F mRNA levels among PV, ET and chronic idiopathic myelofibrosis (IMF) were not significant. Conclusions ARMS-PCR technique can be used to detect the frequency and mutational status of JAK2V617F mutation owing to its sensitivity and along with capillary electrophoresis, quantitative assay for mutated JAK2 mRNA, diagnosis of CMPD and judgement of prognosis become possible.

The clinical and laboratory features of acute promyelocytic leukemia: an analysis of 513 cases / 中华内科杂志

Objective To investigate the clinical and laboratory features of acute promyelocytic leukemia (APL).Methotis 513 APL patients in the last two decades were retrospectively analyzed in this research.We investigated the clinical features including age,sex,abnormality of peripheral hemogram before treatment.therapeutic effect and follow-up and laboratory data such as morphology,immunology,cytogenetics and molecular biology(MICM).Results The median age of the APL patients was 33 years old and the ratio of male and female was 1.21:1.Before treatment,the median level of WBC was 4.3×109/L and the deteetion rate of abnormal promyelocyte on blood film was 85.8%;with immunophenotypie detection,the expression levels of CD117、CD34、HLA-DR、CD7、CD14 and CD19 in APL were found to be lower and the expression 1evels of CD2、CD33 and MPO higher than those in other subtypes of acute myelocytie leukemia(AML)(beth P<0.01).Specific abnormal chromosome t(15;17)was detected in 91.7%of the patients,of whom 75.9%had standard translocation of t(15;17),being the most common one and 15.8% of the patients had t(15;17)with additional abnormal chromosome.There was only 7.5%of the patients with nolnlal karyotype.However,the presence of both simple translocation and complex translocation was seldom seen.With molecular biological detection.PML/RARα fusion gene positive rate was 99.6%.In a relativelv long clinical follow-up,we found that the complete remission(CR)rate in APL patients was 84.7%.incidence of DIC was 13.4%and five-year survival rate was 30.7%.111e median count of WBC in CR group was lower than that non-remission group(P<0.01).There were no significant differences on expressions of CD34 and CD2 and changes of cytogenetics between the two groups(P>0.05).Conclusions Comprehensive evaluation of MICM could be of important significance in the diagnosis and prognosis iudgrnent for APL patients.The CR rate in these patients with high WBC eount was considerable low.

Abnormalities of chromosome 17 in myeloid malignancies with complex chromosomal abnormalities / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the characteristics of the abnormalities of chromosome 17 in myeloid malignancies with complex chromosomal abnormalities (CCAs).</p><p><b>METHODS</b>Abnormalities of chromosome 17 were analyzed in 73 patients with myeloid malignancies with CCAs showed by R banding and conventional karyotyping, including 21 acute myeloid leukemia (AML), 36 chronic myeloid leukemia (CML) and 16 myelodysplastic syndrome (MDS). All CCAs were further analyzed by multiplex fluorescence in situ hybridization (M-FISH).</p><p><b>RESULTS</b>Among the 73 myeloid malignancies with CCAs, chromosome 17 was the most frequently involved chromosome. It was found in 46.5% (34/73) of all cases, including 12 AML, 13 CML in blast crisis (BC) and 9 MDS. However, it was not found in the 9 CML cases in chronic phase (CP). The majority of changes were structural rearrangements which were identified in 43.8%(32/73)of all cases, among them the frequency was 52.4% (11/21), 33.3% (12/36) and 56.3% (9/16) in AML, CML and MDS, respectively. Numerical abnormalities were detected in 15.1% (11/73) cases, all were monosomy 17, and the frequency was 25.0% (3/12), 38.5% (5/13) and 33.3% (3/9) in AML, CML and MDS, respectively. Both numerical and structural abnormalities of chromosome 17 were found in 9 cases. Unbalanced translocations involving chromosome 17 were much more frequent than balanced ones. In the 3 groups, 16, 15 and 8 unbalanced translocations were found respectively. Only two kind of balanced translocations including t(15;17) in AML and t(15;17;22) in CML were found. All chromosomes were involved except chromosomes 5, 6 and 22 as partner chromosomes, the most common one was chromosome 15 (8.2%), followed by chromosome 2 (5.4%). Five of the 6 cases with translocation of chromosomes 15 and 17 were acute promyelocytic leukemia, the other case was CML-BC.</p><p><b>CONCLUSION</b>Abnormalities of chromosome 17 were the most frequently involved chromosomal aberrations in myeloid malignancies, and structural rearrangements were more common. All the numerical abnormalities were monosomy 17, unbalanced translocations were much more frequent than balanced ones.</p>

Simultaneous presence of ins (15;17),t(2;17;20) and trisomy 8 in a patient with acute promyelocytic leukemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To report a rare complex karyotypic abnormalities including ins (15;17),t(2;17;20) and trisomy 8 in a patient with acute promyelocytic leukemia (APL).</p><p><b>METHODS</b>Chromosomes were prepared after 24 h culture of bone marrow cells. R-banding technique was used to analyze the karyotype. Multiplex fluorescence in situ hybridization (M-FISH), chromosome painting using whole chromosome parint (WCP) 2, 15, 17 and 20 and interphase-FISH (I-FISH) using PML-RARa dual-colour dual-fusion translocation probe were performed to ascertain the essence and origin of the abnormal chromosomes detected by conventional karyotypic analysis.</p><p><b>RESULTS</b>Karyotypic analysis revealed a karyotype of 47, XY, 2q-, + 8, 17q+ , 20p+ . M-FISH analysis showed a karyotype of 47, XY, t(2;17;20) (q24;q21;p13), + 8, which was confirmed by chromosome painting. PML-RARa fusion gene which lied in the derivative chromosome 15 was detected by I-FISH suggesting a cryptic insertion (15;17)(q22;q21.1q21.3).</p><p><b>CONCLUSION</b>FISH is a reliable method for characterization of cryptic ins (15;17) and other complex translocations. It should be used in all suspected APL patients lacking t(15;17) by conventional karyotypic analysis.</p>

Fluorescence in situ hybridization detected minimal residual disease and chimerism in patients with hematologic malignancies after allogeneic hematopoietic stem cell transplantation / 中华器官移植杂志

Objective To explore the minimal residual disease (MRD) and cellular chimerism in patients with hematopoietic malignancies after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods From May 2001 to June 2005,83 patients received allo-HSCT,including 55 males and 28 females. Of them 49 patients received sibling HLA-matched bone marrow transplantation (BMT),3 HLA-matched peripheral blood stem cell transplantation,8 un-related BMT,9 nonmyeloablative stem cell transplantation (NST) and 14 related haploidentical transplantation. Among them,49 patients were diagnosed as having CML,16 having AML,16 having ALL,one having multiple myeloma and one having malignant lymphoma. Chimerism and MRD were monitored by fluorescence in situ hybridization (FISH) using X and Y specific centromeric probes or gene probes for BCR/ABL,MLL and AML1/ETO. 1000 cells were analyzed for each sample.Results Among 19 patients receiving sex-matched transplant,the former chromosome rearrangements were not found in 16 patients after transplantation,MRD was detected in 10 % of cells in one patient and 1 % of cells having MRD in 4th month after the reduction of immunotherapy,and the patients were still in remission one year after transplantation. Two patients were found having the former chromosomal rearrangement 1 and 4 months after transplantation,respectively,who did not achieve remission after chemotherapy. Over 99 % donor chimerisms were found in 50 patients on day 25,donor cells were at a low level ( 96.2 % ~ 98.7 % ) in 7 patients on day 25,and increased over 99 % later. They were in remission without relapse. The donor chimerisms were decreased gradually in other 7 patients,of them 3 patients with the host cells above 10 % showed hematologic relapse. Four patients with the host cells between 2 %~5 % had different outcomes: 2 patients died of severe GVHD after the reduction of cyclosporine A,one patient got a donor chimerism over 99 % after reduction of immunotherapy,and one patient was still in complete remission.Conclusions FISH could play a pivotal role in the detection of MRD and chimerism. It is helpful to the evaluation of graft and relapse and to the guide of intervention of early immunotherapy.

Clinical and biological characteristics in childhood acute myeloid leukemia with 8;21 translocation / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical and biological characteristics of childhood acute myeloid leukemia(AML)with 8;21 translocation.</p><p><b>METHODS</b>A retrospective analysis including clinical information, cell morphology, chromosome, immunophenotype and molecular biology was performed on 41 cases of childhood t(8;21)AML. The control group included 19 cases of AML without t(8;21) translocation detected during the same period.</p><p><b>RESULTS</b>The 41 cases of t(8;21)AML accounted for 68.3% of 60 continuous childhood AML patients. Among them, classical t(8;21) translocation was seen in 29 cases; variant t(8;21) translocation, simple 8q-, near-tetraploidy characterized by the duplication of t(8;21) translocation each came into view in 2 cases; and cryptic t(8;21) translocation was seen in 6 cases. Thirty seven cases (80.4%) belonged to M2 subtype of AML. Most of them had the morphological changes such as the leukemia cells' indent nucleus with a light stain region of perinucleus, basophilic cytoplasm, differentiation with maturation, megaloblastoid changes and nuclear-cytoplasm imbalance; the high expression of CD13 antigen; and the AML1/ETO fusion transcript in 23 cases examined by reverse transcription-polymerase chain reaction (RT-PCR) assay, including 6 cases with normal karyotype. The difference in complete remission rate between t(8;21) positive patients group and t(8;21) negative patients group was not significant in statistics (82.4% vs 75%, P>0.05). However the difference in recurring rate of the leukemia was statistically significant (10.7% vs 41.7%, P<0.05).</p><p><b>CONCLUSION</b>t(8;21)AML is the most frequent type of childhood AML. It is predominantly associated with M2 subtype of AML and has unique morphological, immunological prognostic features .</p>

Clinical, cytogenetic and dual-color FISH studies on five cases of myelodysplastic syndrome or acute myeloid leukemia patients with 1;7 translocation / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To study the clinical and cytogenetic characteristics of four patients with myelodysplastic syndrome (MDS) and one with acute myeloid leukemia experiencing t (1;7).</p><p><b>METHODS</b>Five patients seen in our hospital from 1992 to 2001 were diagnosed as MDS and acute myelocytic leukemia (AML) according to the French-American-British (FAB) criteria. Chromosomes were prepared using the direct method as well as 24-hour unstimulated cultures of fresh heparinized bone marrow for each subject, while R-banding was used to analyze karyotypes. Dual-color fluorescence in situ hybridization (FISH) using SpectrumRed and SpectrumGreen directly labeled chromosome 1-specific alpha-satellite DNA probe (red) and chromosome 7- specific alpha-satellite DNA probe (green) was performed for three cases.</p><p><b>RESULTS</b>Of the five patients, three had 1;7 translocation due to a long history of exposure to benzene. In three cases, dual-color FISH resulted in three red signals and two green ones, in which one red signal adjoining one green signal in 27.6%, 84% and 18.5% metaphases, respectively.</p><p><b>CONCLUSIONS</b>Exposure to benzene may be the cause for Chinese MDS and AML patients with t (1;7) translocation. The result of dual-color FISH convincingly confirmed that the centromere of the derivative chromosome 7p/1q resulting from 1;7 translocation was made up of centromeres from both chromosomes 1 and 7.</p>

Detection of common chromosome abnormalities in myelodysplastic syndrome with a panel fluorescence in situ hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To evaluate the value of a panel fluorescence in situ hybridization (FISH) in the detection of common chromosome abnormalities in myelodysplastic syndrome (MDS).</p><p><b>METHODS</b>Twenty cases of MDS patients, whose karyotypes were unknown by the FISH examiner beforehand, were analyzed with a panel FISH using YAC248F5 (5q31), YAC938G5 (7q32), CEP8 and YAC 912C3 (20q12) probes to detect the frequently occurring chromosome abnormalities (-5/5q, -/7q-, +8, 20q-) in MDS. Then the results were compared to those of conventional cytogenetics (CC).</p><p><b>RESULTS</b>Among 20 cases, 13 cases were found to carry common chromosome abnormalities by panel FISH (trisomy 8, five cases; -5/5q-, one case; 20q-, five cases; 5q- accompanying 20q-, one case; complex abnormalities, one case). However, on CC examination, only five cases were found to have common chromosomal abnormalities (20q-, four cases; 5q- accompanying 20q-, one case). In addition, trisomy 21, marker chromosome and complex abnormalities comprising -5, -7 and marker chromosomes were seen in one case each, the rest were normal.</p><p><b>CONCLUSION</b>Panel FISH is a useful tool of molecular cytogenetics in the detection of common chromosome abnormalities in MDS.</p>

Clinical and experimental studies on five cases of acute myeloid leukemia with translocation t(16;21)(p11;q22) / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To report five cases of acute myeloid leukemia (AML) with t(16;21)(p11;q22) translocation and the result of chromosome painting analysis on one of them.</p><p><b>METHODS</b>Chromosome specimens were prepared by short-term culture of bone marrow cells. Karyotype analysis was made by R-banding technique. Chromosome painting was performed using whole chromosome probes 16 and 21 in 1 case.</p><p><b>RESULTS</b>Karyotype analysis showed identical translocation t(16;21)(p11;q22) in all five cases, accounting for 0.3% of 1448 cases of acute myeoid leukemia examined in the past fifteen years. Moreover, chromosome painting distinctly demonstrated t(16;21) in one of them. Leukemia blasts did not show hemophagocytosis in all of them.</p><p><b>CONCLUSION</b>t(16;21) translocation is a rare and recurring chromosome rearrangement. It represents a specific type of AML. Chromosome painting technique is a more reliable means for detecting it, compared with the conventional karyotype analysis.</p>

All-trans retinoic acid as a single agent induces complete remission in a patient with acute leukemia of M2a subtype / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To present a special case with the karyotype and molecular marker of acute myeloid leukemia (AML)-M2 who was induced to complete remission by all-trans retinoic acid (ATRA) alone.</p><p><b>METHODS</b>A recently hospitalized young female patient with acute leukemia was initially diagnosed as M3 subtype based on morphological French-American-British (FAB) classification. Karyotype analysis using standard G and R banding techniques and RT-PCR were applied to further define the diagnosis. After primarily cultured bone marrow cells from the iliac aspiration were tested for in vitro induced differentiation, the patient was treated with oral all-trans retinoic acid alone, 60 mg per day until complete remission was achieved. Peripheral blood and bone marrow changes were monitored over the whole treatment course.</p><p><b>RESULTS</b>The characteristic chromosomal aberration for M3, the t(15;17) reciprocal translocation, was not found while a t(8;21) translocation was verified. Furthermore, an amplified product of the AML-1/ETO fusion gene instead of the PML/RAR alpha fusion gene was detected by RT-PCR and the diagnosis was corrected from M3 to M2. Primary cultured bone marrow cells can be fully induced to terminal differentiation after 4 days exposure to ATRA. A hematological complete remission was achieved after 40 days treatment with ATRA as a single therapeutic agent, suggesting an alternative pathway mediating ATRA-induced myeloid differentiation.</p><p><b>CONCLUSION</b>A leukemia patient with a subtype other than M3, such as M2 in this case, may also be induced to complete remission by the mechanism of ATRA-induced terminal differentiation. This implies that there may be a pathway other than PML/RAR alpha fusion gene product which mediates ATRA-induced myeloid maturation in leukemia cells.</p>