Objective To evaluate the significance of dual-color interphase fluorescence in situ hybridization (FISH) in the engraftment estimation and minimal residual disease (MRD) monitoring after sex-mismatched allogeneic hematopoietic stem cell transplantation (allo-HSCT).Method The fluorescence signal of samples from 35 cases in different periods after sex-mismatched allo-HSCT were detected by interphase FISH using chromosome enumeration probes (CEP) X and Y. Results All of 35 patients had been determined to obtain engraftment after allo-HSCT. When the disease relapsed,FISH showed that the percentage of donor chromosomes was decreased and when the disease got remission,the percentage of donor chromosomes increased. When conventional cytogenetics showed 100 % XX or 100 % XY,FISH showed different percentage of host chromosomes.Conclusions The test of dual-color interphase FISH is reliably sensitive and simple for engraftment evaluation and MRD monitoring post HSCT. It is a good complement method to cell morphology and traditional karyotype analysis.

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.

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.

Objective:To explore the effect of the methanol extract of Macrothelypteris oligophlebia on chronic non-bacterial prosta-titis ( CNP) in rats to confirm the active fractions. Methods:The powdered rhizomes of M. oligophlebia were soaked in methanol. The methanol extract was suspended in water and then extracted successively with chloroform and ethyl acetate to obtain chloroform fraction, ethyl acetate fraction and water fraction. Carrageenan-induced CNP in rats was established. The rats were randomly divided into the sham-operated control group, model group, positive control group, methanol extract group, ethyl acetate fraction group, chloroform fraction group and water fraction group. The anti-prostatitis effect was evaluated by the prostate index, and the pathological examination of prostate was performed using HE staining. The levels of interleukin-10 (IL-10), tumor necrosis factor alpha (TNF-α), cyclooxyge-nase-2 (COX-2) and prostaglandin E2(PGE2) were analyzed using ELISA kits. Results:The ethyl acetate fraction group and metha-nol extract group with high flavonoid content could significantly decrease prostate index (P<0. 01) and the levels of IL-10, TNF-α, COX-2 and PGE2(P<0. 05 or P<0. 01), and improve the prostate morphology when compared with the model group, especially with the ethyl acetate fraction group. Conclusion:The rhizomes of M. oligophlebia show promising therapeutic effect on CNP, and the ethyl acetate fraction is the active fraction.

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.

OBJECTIVETo evaluate the four techniques for clonal analysis in the early diagnosis of myelodysplastic syndromes (MDS).

METHODSFour techniques for clonal analysis were performed in bone marrow samples from fifty patients with suspected MDS: (1) Conventional cytogenetics (CC) for clonal chromosomal abnormalities; (2) BrdU-sister chromatid differentiation (BrdU-SCD) for cell cycle analysis; (3) Fluorescence in situ hybridization (FISH) for trisomy 8; (4) PCR-SSCP for N-ras mutation.

RESULTSThe diagnosis of forty-five patients was compatible with FAB criteria of MDS, the other five patients didn't fully meet the FAB criteria. They had either only one lineage dyspoiesis or no any obvious dysplastic features and two of them were diagnosed as suspicious refractory anemia (RA), one as anemia with hypercellular bone marrow and two as chronic aplastic anemia. The results of the four techniques performed in them showed that four patients had clonal karyotype abnormalities, two had prolonged cell cycle, three had trisomy 8 of different proportions, and one had N-ras mutation. Thus, they were all diagnosed as RA.

CONCLUSIONThe untypical MDS patients can be diagnosed early by examination with combining several clonal analysis techniques.

Adolescent ; Adult ; Aged ; Bromodeoxyuridine ; Child ; Child, Preschool ; Chromatids ; Chromosome Aberrations ; Cytogenetic Analysis ; methods ; Female ; Genes, ras ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Middle Aged ; Mutation ; Myelodysplastic Syndromes ; diagnosis ; genetics

OBJECTIVETo report two myelodysplatic syndromes (MDS) patients with t(3; 5) (q25; q34).

METHODSChromosome specimens were prepared by short-term culture of bone marrow cells. Karyotype analysis was performed by R banding technique, chromosome painting (fluorescence in situ hybridization, FISH) by using whole chromosome 3 and 5 probes in case 1.

RESULTSThe clinical and hematological findings were compatible with diagnosis of MDS. Karyotype analysis showed that both patients had identical t(3; 5) (q25; q34) translocation. A reciprocal translocation between chromosomes 3q and 5q was proved by FISH in one patient.

CONCLUSIONSt(3; 5) translocation is a rare chromosome abnormality specifically associated with MDS and frequently displays trilineage dysplasia. Chromosome painting technique is a reliable tool for detecting this translocation.

Adolescent ; Adult ; Antigens, CD ; analysis ; Chromosomes, Human, Pair 3 ; Chromosomes, Human, Pair 5 ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Leukocytes, Mononuclear ; classification ; immunology ; Male ; Myelodysplastic Syndromes ; genetics ; physiopathology ; Translocation, Genetic

OBJECTIVETo evaluate the association between isolated trisomy 11 and the clinical, hematological, immunological, prognostic aspects in hematological malignancies.

METHODSBone marrow cell cytogenetic analysis was performed by direct method and/or 24 h culture method. RHG banding was used for karyotype analysis. Immunophenotype analysis was carried out by flow cytometry. Ten patients with acute myeloid leukemia (AML) were treated with HA regimen chemotherapy and followed up.

RESULTSThe isolated trisomy 11 was found in 11 of 1 * ! 763 hematological malignancies cases (0.6%). The diagnoses included 10 AML (6 M(2), 2 M(5), 1 M(1), 1 M(4)), and 1 myelodysplastic syndromes. Ten of them have no hepatosplenomegaly. The immunophenotypical analysis of leukemia cells showed positive for CD(13), CD(33) and CD(34) in 5 cases. Follow-up data were available in 10 cases. The complete remission rate was 40% with a median survival of 10 months.

CONCLUSIONThe isolated trisomy 11 was mainly seen in AML, especially in M(2) subtype. Their prognosis was poor.

Adult ; Aged ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; CD13 Antigens ; analysis ; Chromosomes, Human, Pair 11 ; genetics ; Female ; Hematologic Neoplasms ; genetics ; immunology ; pathology ; Humans ; Karyotyping ; Male ; Middle Aged ; Sialic Acid Binding Ig-like Lectin 3 ; Survival Analysis ; Trisomy

OBJECTIVETo evaluate fluorescence in situ hybridization (FISH) in the detection of inv (16) (p13; q22).

METHODSSpectrum red labeled yeast artificial chromosome (YAC) clone 854E2 which spans the breakpoint cluster region in MYH11 in band 16p13 and single color interphase FISH were used to detect inv (16) in 26 cases of acute myelomonocytic leukemias (AML-M(4)), and the results were compared with that of conventional cytogenetic analysis.

RESULTSR banding karyotyping test revealed no inv (16) in 25 cases, one AML M(4Eo) case showed inv (16) by G banding. Nine cases including all three M(4Eo) had inv (16) by FISH analysis, among whom the characteristic fluorescence signal pattern of the inv (16) was seen in 13.3% to 32.1% (median, 21.3%) of the tested cells.

CONCLUSIONYAC 854E2 and interphase FISH provide a powerful technique in the detection of inv (16) (p13q22).

Adult ; Aged ; Chromosome Inversion ; Chromosomes, Human, Pair 16 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Interphase ; genetics ; Leukemia, Myelomonocytic, Acute ; genetics ; pathology ; Male ; Middle Aged

OBJECTIVETo study the clinical and cytogenetic characteristics of four patients with myelodysplastic syndrome (MDS) and one with acute myeloid leukemia experiencing t (1;7).

METHODSFive 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.

RESULTSOf 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.

CONCLUSIONSExposure 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.

Adult ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 7 ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myeloid, Acute ; genetics ; Male ; Myelodysplastic Syndromes ; genetics ; Translocation, Genetic