ZNF217 expression correlates with the biological behavior of human ovarian cancer cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the correlation of ZNF217 gene expression with the biological behavior of human ovarian cancer HO-8910 cells.</p><p><b>METHODS</b>The expression of ZNF217 in ovarian carcinoma cell lines was detected by RT-PCR and Western blot, respectively. The biological behaviors of the transfectants were investigated by MTT, in vitro Boyden chamber and in vivo invasion assay, respectively.</p><p><b>RESULTS</b>RT-PCR and Western blotting revealed that transfection of ZNF217 into the HO-8910 cells significantly increased their proliferation along with markedly enhanced in vitro and in vivo invasion and metastatic abilities. MTT assay showed that the proliferation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells and HO-8910 cells (P < 0.001). The Boyden chamber assay showed that the numbers of migrating pEGFP-N1-ZNF217/HO-8910, pEGFP-N1/HO-8910 and HO-8910 cells were (141.25 ± 13.91) cells/200×field, (82.50 ± 11.73) cells/200×field and (81.75 ± 12.12)cells/200×field, with a significant difference between them (F = 29.247, P < 0.001). The nude mouse experiment showed that the in vivo tumor formation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells (P < 0.001).</p><p><b>CONCLUSIONS</b>ZNF217 gene plays an important role in the invasion and metastasis of ovarian cancer. ZNF217 gene expression may be a useful marker indicating invasion and metastasis of ovarian cancer.</p>

Application of multiprobe fluorescence in situ hybridization panel in the diagnosis of acute myeloid leukemia / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the value of multiprobe fluorescence in situ hybridization (FISH) panel in the diagnosis of acute myeloid leukemia (AML).</p><p><b>METHODS</b>The multiprobe AML/MDS panel comprising 8 different FISH probes for AML1/ETO transfusion gene, PML-RARα transfusion gene, CBFβ/MYH11 transfusion gene, MLL breakapart, P53 deletion, Del(5q), -7/Del(7q), and Del(20q) was tested in 40 cases of AML, and the results were compared with those by conventional cytogenetic G-banding (CCG) test.</p><p><b>RESULTS</b>With multiprobe FISH panel, 22 of the 40 AML cases were found to carry 7 types of cytogenetic abnormalities, namely AML1/ETO transfusion gene, PML-RARα transfusion gene, MLL breakapart, P53 deletion, Del(5q), -7/Del(7q) and trisomy 8. The positive ratio of the multiprobe FISH was 57.5%. CCG only identified 8 cases with the corresponding cytogenetic abnormalities and 3 cases with other cytogenetic abnormalities, and the positive ratio was only 27.50%.</p><p><b>CONCLUSION</b>Mutiprobe FISH panel is more rapid, accurate and effective than CCG in the diagnosis of AML.</p>

Rapid detection of chromosome number aberration in spontaneously aborted fetuses using fluorescence in situ hybridization / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the value of fluorescence in situ hybridization (FISH) in the diagnosis of common chromosome number aberration in spontaneously aborted fetuses.</p><p><b>METHOD</b>A total of 100 spontaneously aborted fetuses were analyzed by G-banding and by FISH to test chromosome number aberration mainly for chromosomes 13, 18, 21, X and Y, and the results of FISH test was assessed according to those by G-banding test.</p><p><b>RESULTS</b>FISH results were well consistent with those by G-banding test. FISH test identified trisomy in 32 samples and polyploidy in 7 samples. Two samples with cell culture failure were found to have trisomy 16 by FISH. Discrepancies in the results between the two tests occurred in 3 samples, but the results of FISH were verified by other methods. Kappa test between FISH technology and G-banding showed a good consistency between FISH and karyotyping (P<0.05).</p><p><b>CONCLUSION</b>FISH is an effective and rapid method for detecting chromosome number aberration in spontaneously aborted fetuses, and the combination of FISH and karyotyping provides more reliable diagnostic evidence.</p>

Minimal residual disease monitoring in chronic myeloid leukemia patients after allogeneic hematopoietic stem cell transplantation using interphase fluorescence in situ hybridization and real-time quantitative reverse transcription PCR / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Interphase fluorescence in situ hybridization (FISH) and real-time quantitative reverse transcription PCR (RQ-PCR) are the common methods for monitoring minimal residual disease (MRD) in chronic myeloid leukemia (CML) patients. This study was to assess the value of monitoring BCR-ABL fusion gene level in CML patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) using FISH and RQ-PCR.</p><p><b>METHODS</b>BCR-ABL fusion gene levels were detected in the bone marrow of 31 patients with CML before and 3-48 months after allo-HSCT using FISH and RQ-PCR.</p><p><b>RESULTS</b>BCR-ABL positive cells detected by FISH were decreased 3-30 months after allo-HSCT and BCR-ABL/ABL mRNA was reduced by 2 logarithmic units in RQ-PCR (P < 0.05). While no BCR-ABL positive cell was detected by FISH 30 months after allo-HSCT, BCR-ABL/ABL mRNA was detected by RQ-PCR and declined by more than 3 logarithmic units, (P < 0.05).</p><p><b>CONCLUSIONS</b>Dynamic monitoring of BCR-ABL gene on molecular level in CML patients after allo-HSCT is useful in the early prediction of susceptibility to recurrence in the patients and in designing intervention, and is thus helpful in improving the overall survival rate after transplantation.</p>

Clinical application of SYBR Green I real-time fluorescence quantitative PCR for diagnosis of common chromosome aneuploidy / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the value of real-time fluorescence quantitative PCR in the diagnosis of chromosome anepuploidy.</p><p><b>METHODS</b>ABCC4 gene on chromosome 13, TYMS gene on chromosome 18, DSCR3 gene on chromosome 21, HPRT2 gene on chromosome X, and SRY gene on Y chromosome were used as the target genes, with GAPDH gene on chromosome 12 as the control gene. Using double-standard curve fluorescent relative quantitative PCR method with SYBR Green as the fluorescent dye, the gene expression levels were detected and the results were compared with those of karyotype analysis.</p><p><b>RESULTS</b>The ratio of the target gene on chromosome 13 to the control gene showed a significant difference between the normal karyotype group (0.90 - or + 0.31) and trisome group (1.39 - or + 0.12, P=0.003), and the genes on chromosome 18 (1.07 - or + 0.44 vs 1.66 - or + 0.12, P=0.000) and chromosome 21 (0.84 - or + 0.27 vs 1.73 - or + 0.54, P=0.000) showed similar results. The expression of the genes on the X chromosome showed no significant difference between 45, X group and 46,XY group (0.62 - or + 0.12 vs 0.63 - or + 0.25, P=0.965), nor between 46, XX group and 47,XXY group (1.32 - or + 0.37 vs 1.20 - or + 0.35, P=0.326), while a significant difference was noted between the single copy X (including 45,X and 46,XY) and two copies X (46,XX and 47,XXY) (0.63 - or + 0.23 vs 1.26 - or + 0.36, P=0.000). The expression of the target gene on the Y chromosome was not detected in normal females (46,XX), and a significant difference in the expression was found between normal male group (46,XY) and 47,XYY group (1.57 - or + 0.54 vs 3.08 - or + 0.15, P=0.003).</p><p><b>CONCLUSION</b>SYBR Green I real-time fluorescence quantitative PCR can be used for the purpose of rapid diagnosis of chromosome aneuploidy.</p>

Simultaneous analysis of telomere length and cell surface antigen in leukemia by multicolor Flow-FISH / 中国实验血液学杂志

This study was purposed to explore the feasibility of simultaneous analysis of telomere length and cell surface antigen by multicolor Flow-FISH to assess minimal residual disease (MRD) in leukemia. The telomere length in 34 leukemia patients versus 20 normal controls was compared by using Flow-FISH, and the relationship between telomere length and therapeutic effect and prognosis was analyzed preliminarily. As for those patients with follow-up samples, the changes of telomere length combined with surface antigen in different courses of disease were observed by multicolor Flow-FISH. The results indicated that the telomere length of de novo patients was significantly shorter than that of controls except the patients in chronic myeloid leukemia-chronic phase (CML-CP). The shorter telomere, the lower complete remission (CR) rates were observed in acute leukemia cases and the shorter duration of CP before onset of blast phase (BP) occurred in CML cases. The acute leukemia patients showed longer telomere and fewer cells expressed the related antigen after CR. The telomere length of cases with continued CR remained at normal level during remission, and there was no increased expression of the specific antigen. However, the telomere of relapsed cases shortened again after relapse with elevated specific antigen expression. In the relapsed cases, the telomere of related antigen positive cells shortened ahead of telomere length change of the whole cells and morphologic change of bone marrow cells. It is concluded that analysis of telomere length by flow-FISH manifests the significance for monitoring disease conditions, estimating prognosis and guiding therapy in all kinds of leukemia. The simultaneous analysis of telomere length and cell surface antigen by multicolor flow-FISH may monitor abnormal clone or clonal evolution to predict recurrence more sensitively and specifically, and may provide a promising and widely applicable method for monitoring MRD in leukemia.

Identification of acute lymphoctic leukemia extramedullary relapse and PTLD after allo-HSCT by monitoring sex chromosome chimeric status with FISH / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the role of monitoring sex chromosome chimeric status by fluorescence in situ hybridization (FISH) in the identification of leukemic extramedullary relapse and post-transplant lymphoproliferative disease (PTLD) in acute lymphocytic leukemia (ALL) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>Six ALL patients who received sex-mismatched allo-HSCT and manifested extravisceral lymphadenectasis or local lump were investigated. The sex chromosome chimeric status in tumor tissues and bone marrows (BM) were monitored by FISH, and EBV-RNA in the tumor tissues were detected by in situ hybridization (ISH).</p><p><b>RESULTS</b>The sex chromosomes in BM of all 6 patients were 100% donor-derived. Among the sex chromosome chimeric status of tumor tissues, three patients were mainly recipient-derived, and the percentage of sex chromosomes derived from recipients were 100%, 100% and 98.0%, respectively, and then they were diagnosed leukemic extramedullary relapse. The other 3 patients were donor-derived, the percentage was 98.5%, 96.0% and 91.5%, respectively, and were diagnosed PTLD. EBV-RNA and latent membrane protein (LMP-1) were positive in 2 patients with PTLD and negative in the other 4 patients. One patient with extramedullary relapse obtained partial remission, one with PTLD gained complete remission, and the others died eventually after therapy.</p><p><b>CONCLUSION</b>Monitoring the sex chromosome chimeric status by FISH is an effective method to distinguish leukemic extramedullary relapse from PTLD in ALL received sex-mismatched donor HSCT.</p>

CD56 and CD11b antigen expressions in patients with acute monocytic leukemia and the clinical implications / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expressions of cell surface differentiation antigen CD56 and CD11b antigen in acute monocytic leukemic (AML-M(5)) cells and their clinical significance.</p><p><b>METHODS</b>A total of 113 cases of de nove adult AML-M(5) were examined genetically and immunologically using G-banding technique, interphase fluorescence in situ hybridization (I-FISH) and flow cytometry immunophenotyping, and the results were analyzed in relation to their clinical data.</p><p><b>RESULTS</b>Of the 113 cases, the expression rates of CD56 and CD11b was 28.32% and 73.45%, respectively. The CD56(+) patients had high CD11b expression, and the expression levels of CD11b and CD56 were positively correlated (P<0.05). The incidence of karyotypic abnormalities was 48.57% (55 cases) in these patients, including 25 (22.12%) with 11q23 aberrations. Twenty-five cases were positive for MLL gene abnormalities as found by I-FISH analysis. Compared with the patients positive for both CD56 and CD11b, those negative for both CD56 and CD11b showed increased peripheral blood white blood cell (WBC) count and also increased blast and progenitor cells in the bone marrow (P<0.05); the former patients often had karyotypic abnormalities, commonly involving 11q23 aberrations (P<0.05), whereas the latter patients presented more likely with extramedullary infiltration and refractory leukemia (P<0.01) with lowered complete remission rate and shortened median survival time (P<0.01). CD56-positive patients were more likely to have karyotypic abnormalities and refractory leukemia than CD11b-postive patients (P<0.05), but the peripheral blood WBC counts, bone marrow blast and progenitor cells, extramedullary infiltration, complete remission rate or median survival time showed no significant differences between them (P>0.05).</p><p><b>CONCLUSION</b>AML-M(5) patients with CD56 positivity and high expression of CD11b often have aberrant karyotypes, commonly involving 11q23/MLL gene abnormality. These patients frequently develop extramedullary infiltration and refractory leukemia often with poor prognosis.</p>

Clinical characteristics and outcomes of 59 patients with acute lymphoblastic leukemia positive for BCR/ABL / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the clinical characteristics and outcomes of BCR/ABL-positive acute lymphoblastic leukemia (BCR/ABL360888725-ALL) and screen the prognostic factors for BCR/ABL360888725-ALL.</p><p><b>METHODS</b>From January 2001 to May 2008, 59 patients (median age of 32 years ranging from 3 to 69 years) with the diagnosis of BCR/ABL360888725-ALL by fluorescence in situ hybridization received induction chemotherapy with VDLP-/+Ara-C regimen. The patients who failed to respond to the chemotherapy received subsequent consolidation chemotherapy with imatinib (400-800 mg/day) (17 cases) or allogeneic hematopoietic stem cell transplantation (allo-HSCT) (16 cases).</p><p><b>RESULTS</b>Of the 59 patients, 32 (58.3%) achieved complete remission (CR) after the first induction cycle. In patients with peripheral white blood cell (WBC) count <30=10(9)/L, 30-99.9(9)/L and > or =100(9)/L, the CR rates were 75.0% (18/24), 56.3% (9/15) and 26.3% (5/19) (P=0.006), and the overall survival probability of 2 years ( OSs of 2-yrs) was 24.7%, 22.5% and 21.1%, respectively (P=0.180). According to the FAB classification, 56 cases were divided into L1, L2 and biphenotypic acute leukemia (BAL) subgroups, and their CR rates were 66.7% (6/9), 63.2% (24/38) and 22.2% (2/9) (P=0.029), with OSs of 2-yrs of 22.2%, 27.0% and 22.0%, respectively (P=0.623). In terms of immunophenotype grouping by EGIL, the patients with ALL, myeloid antigen-positive ALL and BAL had CR rates of 61.1% (11/18), 60.6% (20/33) and 12.5% (1/8) (P=0.039), and the OSs of 2-yrs of 22.7%, 21.0% and 18.8%, respectively (P=0.643). In 55 patients with known karyotype, the CR rates were 71.4%(5/7), 70.8% (17/24) and 37.5% (9/24) in normal, sole t(9;22) abnormality, t(9;22) with additional abnormalities groups (P=0.046), with the OSs of 2-yrs of 42.9%, 34.0% and 7.3%, respectively (P=0.000). The patients complicated by septicemia had significantly lower OSs of 2-yrs than those without septicemia (0% vs 38.8%, P=0.005). The OSs of 2-yrs were significantly higher in patients with consolidation chemotherapy with imatinib than those without (48.0% vs 11.2%, P=0.001), and allo-HSCT was associated with significantly higher OSs of 2-yrs than exclusive chemotherapy (54.2% and 8.5%, P=0.000).</p><p><b>CONCLUSION</b>BCR/ABL360888725-ALL with WBC> or =100 x 10(9)/L, presence of BAL diagnosed by FAB or FACM, t(9;22) with additional chromosome abnormalities all adversely affect the treatment results, and additional chromosome abnormalities and septicemia are associated with lower OSs of 2-yrs. Imatinib treatment and allo-HSCT can both improve the OSs of 2-yrs of the patients with BCR/ABL(+)-ALL.</p>

Detection of ABL kinase domain point mutations in chronic myeloid leukemia patients receiving imatinib treatment / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the frequency and clinical significance of ABL tyrosine kinase point mutations in chronic myeloid leukemia (CML) patients receiving imatinib treatment.</p><p><b>METHODS</b>Nested reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on 40 bone marrow samples from 23 patients to amplify the ABL kinase domain, followed by direct sequencing and sequence homologous analysis.</p><p><b>RESULTS</b>In the 23 patients analyzed, the ABL domain point mutations was detected in 7 patients who presented with 5 types of nucleotide changes, namely T315I(n=3), Y253H, E255K, F317L and G321W. The incidence of mutations in chronic phase (CP), accelerated phase (AP) and blast phase (BP) was 25.00%, 40.00% and 30.00%, respectively. For 6 of the 7 patients with mutations who were resistant to imatinib before sequencing, the daily drug dose had been increased to 600-800 mg daily for poor response to 400 mg/day imatinib. During the follow-up for 3-6 months, only the patient with F317L achieved major cytogenetic response (MCR), and the patient with Y253H and 1 of the 3 with T315I progressed to BP. The newly diagnosed patient with G321W IN cp achieved a complete hematologic remission and had a significant decrease of the proportion of BCR-ABL-positive cells.</p><p><b>CONCLUSIONS</b>ABL kinase point mutation is an important mechanism of imatinib resistance. The type of mutations is associated with the level of resistance to imatinib, and detection of ABL kinase point mutations by direct sequencing may help estimate the prognosis and plan for therapeutic strategy adjustment.</p>

STI571 enhances the effect of arsenic trioxide and Velcade on bcr/abl+-CD34+ cell proliferation and apoptosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effects of STI571, arsenic trioxide (As2O3) and Velcade, used alone or in combination, on the proliferation and apoptosis of bcr/abl+-CD34+ cells.</p><p><b>METHODS</b>bcr/abl+-CD34+ cells isolated from the bone marrow of patients with chronic myeloid leukemia (CML) were treated for 96 h with STI571, As2O3 and Velcade either alone and in combination, and the cell proliferation and apoptosis were analyzed by CCK-8 assay and flow cytometry. The morphological changes of the apoptotic cells were observed by Hoechst33342 staining and fluorescent microscope. The inhibitory effects of the drugs on normal CD34+ cells were also observed.</p><p><b>RESULTS</b>Low-concentration STI571, As2O3 or Velcade all dose-dependently inhibited bcr/abl+-CD34+ cell proliferation without obvious apoptosis-inducing effects. STI571 at 0.25-2 micromol/L combined with As2O3 at 2.5 micromol/L and with Velcade at 15 nmol/L both significantly increased the cell inhibition and apoptosis rates, showing obvious additive or synergistic effects of the drugs without further enhancement of normal CD34+ cell inhibition.</p><p><b>CONCLUSION</b>Combination with STI571 enhances the effects of As2O3 and Velcade on bcr/abl+-CD34+ cells, suggesting the potential clinical value of this regimen.</p>

Effect of G-CSF on the proliferation and differentiation of bcr/abl(+)-CD34+ cells from CML patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the effect of granulocyte colony-stimulating factor (G-CSF) on the proliferation and differentiation of bcr/abl(+)-CD34+ cells.</p><p><b>METHODS</b>bcr/abl(+)-CD34+ cells were isolated from bone marrow of chronic myelocytic leukemia (CML) patients and were treated with 0, 10, 100, 1000 ng/ml of G-CSF for 48, 96, 144 hs. CD34 cells from normal bone marrow were used as controls. Cell proliferation was determined by trypan blue dye exclusion, cell-cycle and antigen differentiation were determined by flow cytometry and cell morphology was observed under light microscope.</p><p><b>RESULTS</b>The number of bcr/abl(+)-CD34+ cells was increased obviously in all groups. After cultured for 48 and 96 h, the number of bcr/abl(+)-CD34+ cells at G-CSF 10 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P < 0.05) , the number of normal CD34 cells was increased only in the presence of G-CSF. After cultured for 48, 96 and 144 h, the cell number in G-CSF 100 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P < 0.05, P < 0.01, P < 0.01, respectively). After cultured for 144 h, the cell percentages in G0/G1 phase for bcr/abl(+)-CD34+ cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that in G-CSF 0 ng/ml group (P < 0. 05), and that for normal CD34 cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that of G-CSF 0 ng/ml group after cultured for 48 and 96 h. The expressions of CD34 on bcr/abl(+)-CD34+ cells and normal CD34+ cells were decreased along with the culture duration, accompanied by the expression of CD33 and CD13 increased first and decreased later, which was not correlated with the concentration of G-CSF. Both bcr/abl(+)-CD34+ cells and normal CD34+ cells showed mature morphology along with proliferation and differentiation.</p><p><b>CONCLUSIONS</b>G-CSF promotes proliferation of both bcr/abl(+)-CD34+ cells and normal CD34+ cells, but not necessary for the former, and the former differentiates more rapidly than the latter does, but both was independent of G-CSF.</p>

ZNF217 gene was detected in ovarian serous cystadenocarcinoma by fluorescence in situ hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate amplification of zinc finger protein 217 (ZNF217) gene in ovarian serous cystadenocarcinoma and its clinical significance.</p><p><b>METHODS</b>Twenty three specimens of ovarian carcinoma, 10 specimens of ovarian benign tumors and 7 specimens of normal ovaries and two ovarian cancer cell lines, SKOV3 and HO-8910 were examined by fluorescence in situ hybridization (FISH).</p><p><b>RESULTS</b>The amplification of ZNF217 was gained in 12 case of ovarian cancers, there was only 1 case in ovarian benign tumor and not amplication in normal ovary.</p><p><b>CONCLUSION</b>The amplification of ZNF217 is associated with ovarian cancer. Oncogenes ZNF217 maybe play a role in cell differentiation and indicate poorer survival in patients with ovarian cancer.</p>

Oncogene ZNF217 amplification on chromosome 20 q in ovarian serous cystadenocarcinoma and its clinical implications / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the amplification of zinc finger protein 217 (ZNF217) gene on chromosome 20 in ovarian cancer and its clinical significance.</p><p><b>METHODS</b>Twenty-three specimens of ovarian carcinoma (11 cases of early stage and 12 advanced stage), 10 specimens of benign ovarian tumors and 7 normal ovaries were examined for ZNF217 gene amplification on chromosome 20 by fluorescence in situ hybridization (FISH).</p><p><b>RESULTS</b>ZNF217 gene amplification was detected in 12 cases of ovarian cancer (52.17%) and 1 case of benign ovarian tumor, but not in normal ovary. ZNF217 amplification was significantly associated with ovarian cancer.</p><p><b>CONCLUSION</b>Oncogene ZNF217 is associated with the tumor stage and poor prognosis of patients with ovarian cancer.</p>

Experimental study of optimized H-2 haploidentical hematopoietic engraftment for the treatment of murine acute leukemia model / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the critical dose of T lymphocyte for preserving graft versus leukemia (GVL) while preventing GVHD in murine acute leukemia model treated with H-2 haploidentical hematopoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>(C57BL/6 x 615) F1 (H-2bk) mice which was inoculated with L615 cells to develop leukemic murine model was the recipient. The healthy C57BL/6 (H-2b) mice was the donor. CD(34)(+) cells from bone marrow and CD(3)(+)cells from spleen of the donor were purified by miniMACS. The purity of CD(34)(+) cells and CD(3)(+) cells were (81.5 +/- 2.4)% and (95.4 +/- 2.9)% respectively. Sixty-nine recipient mice were divided into seven groups. Group A received no treatment, group B received TBI only, the rest groups were irradiated 9 Gy by (60)Co and transfused 10(5) CD(34)(+) cells or mixed with 10(7) (E), 10(8) (F), 1.5 x 10(8) (G) of CD(3)(+) cells respectively. The mice were raised for 60 days, The cause of death was identified by pathology.</p><p><b>RESULTS</b>All mice in group E survived more than 60 days being significantly longer than that in the rest groups (p < 0.0001). The chimerisms from donor were 100% in the mice survived > 60 days. Mice died of leukemia relapse in group D and group E were significantly less than those in group C (p < 0.001). Mice died from GVHD in group G were significantly more than those in group E and group F (p < 0.001).</p><p><b>CONCLUSIONS</b>The leukemia relapse rate was highest in mice that were transplanted with CD(34)(+) cells alone. Those mice transfused with CD(3)(+) T lymphocyte in the graft higher than 10(8) cells died from the GVHD was significantly higher. The inclusive dosage of 5 x 10(7) CD(3)(+) T lymphocyte was enough to separate the GVHD from GVL.</p>

Detection of bcr/abl fusion gene in chronic myeloid leukemia by fluorescence in situ hybridization and nested reverse transcription polymerase chain reaction / 中华血液学杂志

<p><b>OBJECTIVE</b>To detect the bcr/abl fusion gene in chronic myeloid leukemia (CML) for assisting in clinical diagnosis.</p><p><b>METHODS</b>Fluorescence in situ hybridization (FISH) and nested reverse transcription polymerase chain reaction (RT-PCR) were used.</p><p><b>RESULTS</b>Out of 16 CML patients, the results of FISH was not consistent with that of nested RT-PCR in 3 cases. When compared with the results of Northern blot, it was found that nested RT-PCR was more sensitive than FISH, but might give false positive results. Moreover, FISH was the easier method for detecting rarer types of bcr/abl transcripts. For detecting the latter occasions nested RT-PCR have to design several specific primers.</p><p><b>CONCLUSION</b>It could take their advantages of applying FISH and nested RT-PCR at the same time to make the results more accurate and reliable for the diagnosis, prognosis and monitor of minimal residual disease.</p>