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Purpose of this study was to analyse the characteristics of clinical, iconographical, pathological and treatment methods of Langerhans cell histiocytosis (LCH), so as to improve the diagnosis and treatment level of this disease. The clinical data of 35 LCH patients were studied retrospectively. These patients were divided into 2 groups according to age <14 years old and ≥ 14 years old. The clinical symptoms were analysed and the signs, imageology and pathology manifestation and treatment results were evaluated. The results showed that LCH clinical manifestations were diverse and complex. Surgical treatment for patients with single system involvement of LCH was better than that of multi-system involvement of LCH (MS-LCH). For the latter, combined chemotherapy effects was better. After 3-year follow-up, 1-year OS was 94% ± 4%, 2-years OS was 91% ± 5%, 3-year OS was 86% ± 7%. 3 years OS of group <14 years old and ≥ 14 years old was 94% ± 6% and 81% ± 10% respectively. The OS of former was better than that of the later, but because a small number of cases, this difference was not statistically significant. It is concluded that LCH is easy to be misdiagnosed, the pathological biopsy is the gold standard of LCH diagnosis. The PET-CT can be of great help in identifying stages and finding lesion areas of the disease. Pulmonary Langerhans cell histiocytosis (PLCH) is more common in adult. Combined chemotherapy can improve the prognosis of the patients. The treatment methods should be choosed according to the stage and classification of disease.

This study was aimed to detect the expression of AML1 fusion genes in the patients with adult acute myeloid leukemia (AML) and further to investigate their association with the progression and prognosis of AML. Bone marrow samples were collected from 168 patients with de novo adult AML, and the expression of AML1 ETO, AML1-EVI1, AML1-MDS1, AML1-MTG16, AML1-PRDM16, AML1-LRP16, AML1-CLCA2 and AML1-PRDX4 was analyzed by a novel multiplex nested RT-PCR. Positive samples and minimal residual disease were further examined by real-time fluorescent quantitative PCR. The results showed that the AML1 fusion genes were found in 10.7% (18/168) patients. Among them, AML1-ETO in 12 (7.1%) cases were detected, AML1-EVI1 in 2 cases (1.2%), and AML1-MDS1, AML1-MTG16, AML1-PRDM16, and AML1-CLCA2 in 1 case (0.6%) each were detected. Among the patients with AML1-ETO, 10 patients (10/12, 83.33%) achieved complete remission (CR) after one cycle of chemotherapy, while 2 patients achieved CR after 2 cycles of chemotherapy. The 2 patients with AML1-EVI1 failed to achieve CR after one cycle of chemotherapy. Patients with AML1-MDS1, AML1-MTG16, AML1-PRDM16, or AML1-CACL2 did not achieve CR after one cycle of chemotherapy. It is concluded that AML1 fusion genes are more frequent and can provide the molecular markers for diagnostics and prognosis evaluation of AML and for monitoring MRD.

This study was purposed to investigate the clinical efficiencies and adverse reactions of treating the myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) by using decitabine. The clinical data of 12 MDS and AML patients treated with decitabine were analyzed retrospectively. Among 12 patients there were 1 case of MDS-RA, 2 cases of MDS-RAEB-I, 3 cases of MDS-RAEB-II, 2 cases of AML-M4, 2 cases of AML-M5, 1 case of AML-M6 and 1 case of AML-M0. In decitabine chemotherapy program for 5 days (n = 8), decitabine 20 mg/(m(2)·d) × 5 days was applied, 4 weeks for 1 cycle; in program for 3 days (n = 2), decitabine 15 mg/m(2), once 8 h for 3 days, 6 weeks for 1 cycle; another program (n = 2), decitabine 20 mg/(m(2)·d) every other day for 5 times. For 1 patient achieved complete remission (CR) after treatment with decitabine, ID4 gene methylated level was detected by MS-PCR and ML-PCR before and after treatment. The results showed that 2 cases achieved CR, 1 case partial remission, 5 cases stable disease, 1 case progress of disease and 3 cases died. Disease control rate was 66.67% (8/12), the effective rate 25% (3/12). The average survival time was (11.5 ± 2.1) months. 1-year OS rate was 40%, 2-year OS rate was 16.7%. MS-PCR detection showed that the decitabine could significantly reduce the ID4 gene methylation level. It is concluded that decitabine can stabilize disease status of MDS patients, reduce blood transfusion dependence and improve the life quality of patients, and even some patients who transformed from MDS to leukemia achieved CR after treatment with decitabine. Decitabine can reduce the ID4 gene methylation level. The main adverse reaction of decitabine was myelosuppression, infection and so on. So the blood transfusions, antibiotics and other supportive treatments for these patients are needed. Most of patients well tolerate the adverse effects of decitabine after active symptomatic and supportive treatment. The efficacy and survival rate of patients in this study were similar to that of application of decitabine to treat MDS in other domestic studies.

Alteration in the balance between cell apoptosis and proliferation is one of the pathophysiological mechanisms of the myelodysplastic syndromes (MDS). The question of whether the excessive apoptosis and/or proliferation predominantly involve the subset of progenitor cells (CD34(+) cells) or mature cells (CD34(-) cells) remains a controversial issue. This study was purpose to analyze the apoptosis and proliferation status of CD34(+) and CD34(-) cells in bone marrow (BM) of patients with MDS, to investigate the pathogenesis of MDS and to determine the relation of apoptosis and proliferation status of CD34(+) and CD34(-) cells with prognosis of MDS. The proportion of CD34(+) cells, the apoptosis and proliferation ratio (A/P) of CD34(+) and CD34(-) cells in BM of 40 patients with MDS, including 20 cases of high-risk MDS and 20 cases of low-risk MDS, and 10 normal persons as control were detected by flow cytometry; the influence of CD34(+) and CD34(-) cell apoptosis and proliferation levels on prognosis of MDS was evaluated by univariate and multivariate analysis of survival. The results showed that the proportion of CD34(+) cells in BM of high-risk MDS patients was significantly higher than that in BM of low-risk MDS patients and in normal BM [(1.92 ± 0.10)%, (1.09 ± 0.04)%, (1.03 ± 0.05)% respectively]. The apoptotic rates (AR) of both CD34(+) and CD34(-) cells were significantly higher in low-risk MDS [(54.75 ± 2.18)%, (80.36 ± 1.68)%] than in high-risk MDS [(24.87 ± 2.69)%, (23.12 ± 1.23)%] and in normal BM [(18.51 ± 2.74)%, (20.98 ± 2.21)%]. When compared between CD34(+) cells and CD34(-) cells in low-risk MDS, a greater AR of CD34(-) cells was found. However, the higher proliferative rate of CD34(+) cells was observed in high-risk MDS. In low-risk MDS, a higher A/P ratio was found in CD34(-) cells than in CD34(+) cells; whereas this ratio was equalized or inverted in high-risk MDS. In addition, the survival and prognosis correlated significantly with AR of CD34(+) cells. It is concluded that the early MDS is predominantly associated with excessive apoptosis of the mature CD34(-) cells. The proliferation rate of cells increases with the disease progression in MDS subsets, especially, in the subset of CD34(+) cells. Surprisingly, the apoptosis of CD34(+) cells may be a useful prognostic factor, and the inhibition of apoptotic mechanisms may induce the transformation of MDS to leukemia.

Objective: To explore the role of SHP-1 promoter methylation on the pathogenesis and progression in myelodysplastic syndromes (MDS) and its related mechanism., Methods: 63 MDS patients were divided into low-grade (LG) group and high-grade (HG) group according to IPSS score system. Bone marrow samples were collected. Methylation specific-PCR (MSP) were used to detect the status of SHP-1 promoter methylation in bone marrow (BM) samples from different risk MDS patients and MDS cell line, SKK-1. Western blot was used to detect signal transduction and activator of transcription (STAT3) activation in SKK-1 cell line and MDS patients., Results: No SHP-1 promoter methylation could be detected in healthy controls BM. Partially methylation was found in SKK-1 cell line. Methylation rate of SHP-1 gene promoter was found in BM of 24.2% of low-grade MDS patients and 63.3% of high-grade MDS patients, the difference between these two groups was statistically significant (P < 0.05); Patients were divided into different groups according to WHO subtype, chromosomal karyotype and blast cells in bone marrow, methylation rates of SHP-1 were significantly higher in RAEB-II, poor karyotype group and samples with 0.11-0.19 blast cells (P < 0.05); The phosphorylation protein of STAT3 was detected in SKK-1 cell line. The expression of phosphorylation STAT3 was significantly higher in HG group than in LG group (66.7% vs 18.2%) (P < 0.05). There was a significant correlation between SHP-1 promoter methylation and STAT3 phosphorylation., Conclusion: Abnormal methylation of SHP-1 gene promoter might have tentative role in the pathogenesis and progression of MDS, which may be involved in STAT3 activation. Detection of SHP-1 promoter methylation may be helpful to evaluate the prognosis of MDS.

Objective: To evaluate the value of spectral karyotyping (SKY) in cytogenetic analysis of acute myeloid leukemias (AML)., Methods: Nine AML patients were analyzed by R-banding and SKY. MLL, PML-RARalpha, AML1-ETO fusion genes were detected by dual fusion- fluorescence in situ hybridization (D-FISH)., Results: All 9 samples were successfully hybridized. SKY identified structural aberrations including 9q -, t(15;17) and ins(10;17) (q22;p11p12) ; and some numeral abnormalities. The results of SKY confirmed those of R-band karyotyping and D-FISH; with more accurate localization., Conclusion: SKY appears to be fairly stable, accurate and sensitive, for AML cytogenetic study.

This study was aimed investigate the recombination event occurring between HLA-A and-A loci discovered from father's HLA haplotype chromosome in a family. Peripheral blood samples were collected from a family. HLA class I (-A, -B, and -Cw) and II (-DRB1 and -DQB1) alleles were amplified and typed by both low and high resolution PCR with sequence-specific primers (PCR-SSP) and sequence-based typing (SBT). The results showed that 2 haplotypes of the patient were A(*)3001-B(*)1302-DRB1(*)0701 and A(*)3001-B(*)5601-DRB1(*)1454 respectively, those of her father were A(*)3001-B(*)1302-DRB1*0701 and A(*)1101-B(*)5601-DRB1(*)1454. Family analysis demonstrated that the patient's A(*)3001-B(*)1302-DRB1(*)0701 came from her mother and A(*)1101-B(*)5601-DRB1(*)1454 came from her father, but the A of patient was A(*)3001 and B, DR were the same to her father. This showed that the chromosome exchange and recombination event of father's 2 haplotypes occurring between HLA-A and -A loci at meiosis. And recombinate haploid chromosome was completely inherited to his daughter 1. HLA typing and Paternity testing demonstrated that father was the natural father, and the recombination event occurring between HLA-A and -A loci of the daughter 1 with father's HLA haplotype chromosome. It is concluded that the HLA-A/A of father's HLA haplotype chromosome recombination event occurring between HLA-A an-A loci has been found in a family in China, which helps further study on the mechanisms of HLA recombination.

Objective: to investigate the chromosomal characteristics of pancreatic ductal adenocarcinomas by spectral karyotyping., Methods: cytogenetic aberrations of pancreatic cancer cell line P2 established from a Chinese patient was investigated by spectral karyotyping (SKY). Chromosomal alterations were further evaluated in 10 cases of pancreatic cancer and 10 cases of chronic pancreatitis by two color fluorescence in situ hybridization (FISH) by using EGFR/CEP7 probe and paraffin embedded tissue samples., Results: hypotriploid and 26 chromosomal aberrations were revealed in cell line P2. Recurrent chromosomal numerical alterations included loss of chromosome 4, 9, 18, 19, 22, Y, 10p, 15p, 8p, 6q and 12p, with gain of chromosome 7 and 12q. Frequent chromosomal structural abnormalities included der(9;15)(q10;q10), der(10)(3;10)(?;q26) and der(12)(8;12)(?;p13). Four of 10 cases showed EGFR copy number changes by FISH., Conclusions: highly complex chromosomal rearrangements occur in pancreatic cancers. Additional studies of more cases are needed, including FISH analysis of EGFR copy number changes, to reach a conclusion.

This study was aimed to investigate the clinico-pathological features, diagnosis and treatment of the 8p11 (eight p11) myeloproliferative syndrome (EMS). Morphological changes of cells were evaluated by bone marrow smear and biopsy. The cell immunophenotypes were analysed by flow cytometry. Karyotypes were determined by conventional cytogenetic method, and bcr/abl fusion gene was detected by reverse transcription-polymerase chain reaction (RT-PCR). The results indicated that EMS was a relatively rare disease characterized by the occurrence of a bcr/abl-negative myeloproliferative disorder and a T-cell lymphoblastic lymphoma (T-LBL). Bone marrow examination showed myeloid hyperplasia or myeloproliferative neoplasm, often accompanied by eosinophilia. Flow cytometric immunophenotyping showed increased myelomonoblasts; cytogenetic analysis showed a translocation at the 8p11 locus; RT-PCR demonstrated non bcr/abl fusion gene. At the molecular level, all cases carried a chromosomal abnormality involving the fibroblast growth factor receptor 1 (FGFR1) at chromosome 8p11. Up to now, 11 partner genes have been identified and associated with FGFR1 rearrangements. The most common partner is ZNF198 on chromosome 13q11-12. Majority of patients terminate in acute myeloid leukemia which is resistant to conventional chemotherapy. Currently, the only curative option appears to be allogeneic hematopoietic stem cell transplantation. In conclusion, EMS is myeloid and lymphoid neoplasm, associates with FGFR1 rearrangements. It is usually misdiagnosed as T-LBL, atypical chronic myeloid leukemia (aCML) or chronic myelogenous-monocytic leukemia (CMML). Timely cytogenetic and molecular biological examination is vital in order to avoid misdiagnosis and mistreatment.

Objective: To compare the efficacy and adverse effects of bortezomib + adriamycin + dexamethasone (PAD) and vincristine + adriamycin + dexamethasone (VAD) regimens in untreated multiple myeloma (MM)., Methods: There were 26 and 28 new diagnosed MM patients in PAD and VAD groups. Both clinical effects and adverse effects were observed. Patients accepted VAD or PAD regimens for 2 - 4 cycles and followed up for 7 - 27 months., Results: There were 10, 5 and 11 patients accepted 2, 3 and 4 cycles in PAD group, and 6, 11 and 11 in VAD group. In PAD group, there were 2, 14, 9, 1 and 0 patients achieved complete remission (CR), very good partial remission (VGPR), partial remission (PR), stable disease (SD) and progressive disease (PD); in VAD group, the number were 0, 4, 12, 10 and 2. The rate of patients who achieved good efficacy (CR + VGPR) in PAD group was 61.5%, which was higher than that in VAD group (14.3%). The incidences of infection and gastrointestinal symptoms were similar in the two groups, while the incidences of peripheral neuropathy, thrombosis and Herpes Zoster infection in PAD group were higher than those in VAD group., Conclusions: Compared with the conventional VAD chemotherapy, PAD may improve CR and VGPR rates in new diagnosed MM, while it may bring more and severer toxicities in peripheral neuropathy, thrombosis and Herpes Zoster infection. Preventive medical care is necessary in PAD protocol.

It was successful to denoise the spectrum signal within visual wave band (350-560 nm) by wavelet transformation, to extract the folic acid characteristic wavelength 366 nm and some character wavelengths with relation to chlorophyll at 380, 414, 437 and 554 nm. In the range from 560 to 2500 nm, after denoising, the biggest error was smaller than 1.47%, while at the peak or vale of character wavelength the biggest error was smaller than 0.11%. Moreover, the model was established based on the denoised data acquired with aid of plant probe. The model was also based on BP neural network and for predicting the nitrogen content in nutrient solution in hydroponic cultivation of tomato. The results showed that the predicting model, which used the values of absorbance at 554, 673, 1440 and 1940 nm as input variables of BP neural network,had a very good forecasting accuracy and great potential to be used practically.

The study was aimed to evaluate the adverse effects of PAD (bortezomib + adriamycin + dexamethasone) and VAD (vincristine + adriamycin + dexamethasone) as chemotherapy regimens in multiple myeloma patients. 27 and 30 patients with multiple myeloma (MM) were enrolled in PAD and VAD groups respectively. MM patients accepted 3 - 5 cycles of VAD or PAD regimens. The type, degree and occurrence time of adverse reactions during the treatment were observed. The results showed that the rash was found in two patients only in PAD group, leucocytopenia, thrombocytopenia, peripheral neuropathy, infection, fatigue, nausea, constipation, and adverse effects of cortex hormone (hypertension, glycemia, hypokalemia, hyponatremia and acne) were found in the both two groups. The thrombosis was not observed in both two groups during treatment. Although statistical analysis indicated that only the incidence of thrombocytopenia was higher in PAD group than in VAD group with statistical difference but the incidence of leucocytopenia, peripheral neuropathy and infection in PAD group were higher than those in VAD group. Rash, constipation, peripheral neuropathy could be found in the first course of chemotherapy, while the others mostly emerged after 3 courses of treatment. The main reasons for the patients who's treatment was stopped include infection and intolerable peripheral neuropathy. Although peripheral neuropathy could be found in the two groups, but the chemotherapy was stopped only in 2 patients of PAD group after 3 cycles of treatment. It is concluded that compared with conventional VAD chemotherapy, PAD may improve therapeutic effect, but it may bring more severe toxicities to the patients with multiple myeloma.

This study was aimed to compare the efficacy and adverse effects of PD (bortezomib + dexamethasone) and VAD (vincristine + adriamycin + dexamethasone) as regimens for treatment of multiple myeloma patients. 21 and 31 multiple myeloma patients were enrolled in the PD and VAD groups respectively which received 2 to 5 courses of treatments, and both clinical effects and adverse reactions were observed. In the all 52 patients, 48 were newly diagnosed and the other 4 patients had accepted 1 to 2 courses of M2 or MP treatment, but didn't get PR. In 52 patients, 4, 4, 8 and 5 patients accepted 2, 3, 4 and 5 courses of PD regimen respectively, while 6, 11, 12 and 2 patients accepted 2, 3, 4 and 5 courses of VAD regimen respectively. The results indicated that the rate of good efficacy (both CR and VGPR) in PD group was 57.1%, while the rate of good efficacy in VAD group was 16.1%, there was significant difference (p = 0.0052). The percentage of patients who got CR, VGPR and PR in PD and VAD groups were 95.2% and 74.2% respectively, there was no significant difference (p = 0.1108). The incidences of adverse effects in 2 groups were similar, which included hematological toxicity, liver and kidney functional lesion, peripheral neuropathy, infection, interstitial pneumonia. It is concluded that compared with conventional VAD chemotherapy, PD may improve CR and VGPR rate in newly diagnosed patients with multiple myeloma, meanwhile it does not bring about more and worse toxicity.

This study was aimed to explore the value of dual-color dual-fusion fluorescence in situ hybridization (DC-DF-FISH) for the detection of bcr/abl fusion gene in chronic myeloid leukemia (CML). The karyotypes of chromosomes and bcr/abl fusion gene in 41 cases of CML including 18 cases of de-novo CML, 18 treated CML cases and 5 cases of CML received PBSCT were detected by conventional R-banding technique, DC-DF-FISH and RT-PCR. The results indicated that the Ph chromosome was found in 17 out of 18 cases of de novo CML by R-banding technique, with positive rate of 94.4%; DC-DF-FISH detection showed same result (94.4%). The R-banding technique was adopted to detect 18 treated patients and showed that 14 cases had metaphase for analysis, the Ph chromosome existed in 11 out of 14 cases with positive rate of 78.6% (11/14), however, DC-DF-FISH detection also showed positive rate of 94.4% (17/18) for these treated patients. The Ph chromosome in 5 cases after PBSCT did not found by R-banding technique, meanwhile FISH detection indicated that 1 case had bcr/abl gene, RT-PCR assay confirmed the result of FISH detection. It is concluded that the DC-DF-FISH technique is an accurate and reliable method for detecting bcr/abl gene which can be used in diagnosis of CML, evaluation of therapeutic efficacy and detection of minimal residual disease.

Depletion of T and B cells from the graft is prerequisite for haploidentical transplantation to decrease the risk of GVHD and EBV-associated lymphoproliferative disease. This study was aimed to investigate the performance of T-cell and B-cell simultaneous depletion from mobilized peripheral blood stem cells (PBSCs) for the first time in China, using anti-CD3 and anti-CD19 antibodies conjugated to magnetic microbeads by the CliniMACS device. The depletion efficiency of T-cell and B-cells was analyzed by flow cytometry; the function of the stem cells after depletion was evaluated using colony assays. The results indicated that the mononuclear cell count prior to T- and B-cell depletion was 4.88 x 10(10). After depletion, the percentage of T cells was 0.02% with a log (10) depletion of 4.4. The percentage of B cells was less than 0.01% with a log (10) depletion of at least 3.3. The product contained not only CD34(+) stem cells, but also NK cells, monocytes and granulocytes. After T- and B-cell depletion the purity of CD34(+) cells was 0.98%, the number of CD34 cells was 1.84 x 10(8) and their recovery rate was 69.7%. The number of NK cells was 2.54 x 10(9) and the recovery rate of NK cells was 71.7%. In vitro colony assays showed no negative impact on function of the hematopoietic stem cells. In conclusion, the CliniMACS system can be used to efficiently deplete T and B cells from PBSCs simultaneously, without adverse effect on biological function of hematopoietic stem cells. This study provides technical platform for haploidentical hematopoietic stem cell transplantation.

This study was aimed to investigate the chimerism and fusion gene expression in patients with CML after allo-HSCT, to analyse engraftment and minimal residual disease by using STR-PCR combined with RT-PCR qualitative and quantitative assays, and to evaluate their clinical value for predicting disease relapse. 4 relapsed patients with CML after allo-HSCT were dynamically investigated. Qualitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr/abl transcripts was performed by RT-PCR. The results showed that the 100% donor chimerism appeared in 4 patients on day 28 after transplantation and bcr/abl expression was negative, but the 4 patients were in status of unstable mixed chimerism (DC: 0% - 80.4%) at the different time points during the following up with bcr/abl gene positive. 2 patients of them were continuously mixed chimerism after relapse of CML, the other 2 changed from MC to CC by intervention of clinical treatment. Decreasing values of donor chimerism were detected prior to the occurrence of graft rejection and CML relapse, and bcr/abl gene expression was positive. It is concluded that the results of STR-PCR in the range of its sensitivity fully correspond with bcr/abl tests in patients. The combination of STR-PCR with RT-PCR will provide a highly sensitive and valuable tool for evaluating engraftment, graft rejection, and relapse and predicting GVHD. Furthermore, it can provide a basis for early intervention of clinical treatment, and can identify these high risk patients with molecular or cytogenetic relapse after allo-HSCT.

Objective: To evaluate the safety and outcome of patients with acute myocardial infarction (AMI) transferred for primary percutaneous coronary intervention (PCI)., Methods: Data from patients with ST elevation AMI urgently transferred from first admitted hospitals to our cath-lab to receive primary PCI were analyzed. According to time intervals from symptom onset to transfer, the patients were divided into early transfer (< 6 h, n = 26), delayed transfer (6 - 24 h, n = 39) and late transfer (24 h to 1 week, n = 18) group. The major cardiac events during transfer periods and one month after PCI were obtained and echocardiogram and left ventricular systolic functions were compared among groups., Results: There was no serious cardiac event during transfer period and all 83 patients received primary PCI with a mean transfer-to-balloon time about 180 minutes. Success rate of PCI was 92.3% in early transfer group, 89.7% in delayed transfer group, and 94.4% in late transfer group (P > 0.05). At one month follow-up after PCI, 0, 10.3% and 16.7% of patients developed heart failure in early, delayed transfer and late transfer group respectively (P > 0.05 vs. early), the LVEF of early transfer group (53.2% +/- 9.7%) was also significantly higher than delayed transfer group (48.6% +/- 8.2%, P < 0.05) and late transfer group (43.1% +/- 10.3%, P < 0.01)., Conclusions: Transfer patients with AMI for primary PCI is safe in the observed time intervals during acute phase. Early transferred patients are associated with better outcome at 1 month post PCI compared to delayed and late transferred AMI patients.

This study was aimed to evaluate the effects of recombinant human interleukin 11 (rhIL-11) and recombinant human granulocyte colony stimulating factor (rhG-CSF) in mobilization for autologous peripheral blood stem cell transplantation (APBSCT). 16 patients with non-Hodgkin's lymphoma or acute myeloblastic leukemia were given myelosuppressive chemotherapy, then were mobilized by using rhG-CSF 5 microg/(kg.d) for median 5.5 days and rhIL-11 50 microg/(kg.d) for median 4 days (experimental group) or rhG-CSF 5 microg/(kg.d) alone for median 5.5 days (control group). After mobilizing, the peripheral blood leucocyte and platelet counts, total mononuclear cells, CD34+ cells and CFU-GM counts in PBSC collection, and amount of apheresed platelet transfusion were assayed. The results showed that the peripheral blood leucocyte and platelet counts, total mononuclear cell, CD34+ cell and CFU-GM counts in PBSC collection were no significant difference between two groups (p>0.05). After APBSCT, the median time for neutrophil count>or=0.5x10(9)/L and the median time for platelet count>or=20x10(9)/L were 10.5 and 11.5 days in experimental group, while were 13 and 13 days in control group, respectively. The median amount of apheresed platelet transfusion was 3.5 unit in experimental group and 5 unit in control group. Data were significantly different between two groups (p<0.05). The adverse reactions of mobilization were mild fever, fatigue, symptoms like as common cold, poor appetite, dizziness, muscular soreness in experimental group, but only mild fever in control. These symptoms were well tolerated and overcome with drug withdrawal. It is concluded that the regimen of rhIL-11 in combination with rhG-CSF after myelosuppressive chemotherapy to mobilize PBSC is efficient and safe with rapid hematologic reconstitution and less platelet transfusions after APBSCT were used.

The aim of this study was to evaluate the safety and efficacy of allogeneic hematopoietic peripheral blood stem cell transplantation (allo-PBHSCT) for post-operative therapy of acute non-lymphocytic leukemia (ANLL) patient complicated with renal cell carcinoma (RCC). One ANLL patient complicated with RCC underwent an myeloablative HLA-identical relative allo-PBHSCT after RCC operation. The conditioning regimen consisted of total body irradiation, cyclophosphamide and cytarabine. Graft versus host disease (GVHD) prophylaxis regimen composed of cyclosporine A, myco-phenolate mofetil and short course of methotrexate. The results indicated that the patient achieved engraftment 16 days after transplantation with full donor-type chimerism detected by STR-PCR at 30 and 100 days after transplantation. No acute or chronic GVHD and any severe complication developed. As of March 2007, the patient remains without disease at follow-up of 44 months. In conclusion, allo-HSCT procedure is feasible and effective for post-operative therapy of ANLL patient complicated with RCC without severe toxicity.

The objective of this study was to investigate the methylation status of zonula occluden protein-1 (ZO-1) gene in patients with myelodysplastic syndrome (MDS) and to identify its roles in pathogenesis, development and classification of MDS. 85 patients with MDS and 30 healthy individuals were tested by methylation specific polymerase chain reaction (MS-PCR). The results indicated that no ZO-1 promoter methylation could be detected in healthy controls. methylation of ZO-1 gene promoter of bone marrow was found in 56.5% (48/85) MDS patients. The difference between these two kinds of subjects was statistically significant (p<0.05). The methylation status of ZO-1 gene promoter region in the subtypes of MDS was as following: RA (18/37, 48.6%), RAS (4/6, 67%), RCMD (19/30, 63%), RAEB (7/12, 58%). Every subtype of MDS patients had statistical difference from healthy people (p<0.05), but between the subtypes of MDS there were no significant statistical differences in the methylation status of ZO-1 gene, while the level of ZO-1 promoter methylation in group of RA was lower than that in other groups. It is concluded that the ZO-1 promoter region in bone marrow of MDS patient shows a hypermethylation status, which is specific for MDS. MDS is a common hematologic malignancy with clonal proliferation, it is difficult to differentiate from many other hematologic malignancies in clinical diagnosis. However, the change of ZO-1 gene methylation status is closely related to pathogenesis of MDS, therefore the ZO-1 gene as valuable diagnostic marker has important clinical significance. The ZO-1 gene may be a potential gene related to hematologic malignancies.

The aim of this study was to analyze chimerism, evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) by multiple short tandem repeat (STR) amplification using fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis. Peripheral blood and bone marrow in 27 patients who received myeloablative allogenetic cell transplantation were collected before and after transplantation in different times. 10 and 7 different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus/Cofiler plus PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype soft ware were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient. The results showed that donor chimerism was similar by the two methods. The median number of informative alleles was 6.3 (4 - 9) by Profiler Plus and 4.9 (2 - 6) by Cofiler Plus. The donor alleles appeared in 26 patients on day 28 post transplantation. One patient was not observed to appear donor alleles. 14 patients with 100% donor chimerism (DC) had stable engraftment and they still survive in free leukemia. 9 patients had unstable mixed chimerism (DC: 0% - 90.2%), and 5 of them relapsed after allo-HSCT, 6 patients died. Decrease of donor chimerism appeared prior to graft rejection and disease relapse. The incidence of GVHD was higher in group of full donor chimerism. It is concluded that dynamic monitoring donor chimerism by STR-PCR in combination with all auto-capillary electrophoresis is a valuable tool for predicting graft rejection, disease relapse and occurrence of GVHD, and provides a basis for early clinical intervention in the patients received allo-HSCT.

Objective: To establish a spectral karyotyping (SKY) technique and explore the value of SKY in leukemia research., Methods: SKY technique was conducted on 2 samples of peripheral blood from 2 healthy volunteers, then on the samples from 8 patients with leukemia, including chronic myelocytic leukemia (CML) and acute myelocytic leukemia (AML) confirmed by R-banding. In addition, four patients underwent dual fusion-fluorescence in situ hybridization (DF-FISH) to detect the mixed lineage leukemia (MLL), PML/RARa, and BCR/ABL fusion genes. By comparing the results of SKY, R-band karyotyping, and DF-FISH, the stability and reliability of SKY was judged., Results: All 10 samples were successfully hybridized and karyotyped. The 2 cases of healthy volunteers showed normal karyotypes, thus, a specific SKY technique was successfully established. In the 8 cases of leukemia patients, SKY identified aberrations including 9q-, t (9; 22), t (15; 17) and the complex karyotype 47, XY, +9?ins (1;5) (q23;q23), t (6;7) (q23?; p13), in addition, the SKY technique detected some number abnormalities. The results of SKY confirmed the results of R-band karyotyping and DF-FISH; moreover, the SKY technique provided more accurate karyotypes., Conclusion: With high stability, accuracy, and sensitivity, the SKY technique established by this study can be applied in leukemia research.

Spectral karyotyping (SKY) is a novel cytogenetic technique, has been developed to unambiguously display and identify all 24 human chromosomes at one time without a priori knowledge of any abnormalities involved. SKY discerns the aberrations that can not be detected very well by conventional banding technique and fluorescent in situ hybridization (FISH). So SKY is hyper-accurate, hypersensitive, and hyper-intuitional. In this paper the basic principle of SKY technique and its application in leukemia cytogenetics were reviewed.

This study was aimed to explore feasibility and efficacy of unrelated donor peripheral blood stem cell transplantation (UD-PBSCT) in treatment of hematologic malignancies. Ten patients with hematologic malignancies underwent high resolution DNA based typing HLA-matched or 1 locus mismatched UD-PBSCT. Busulfan, cyclophosphamide, Ara-C, MeCCNU and antithymocyte globulin (ATG) were used for preconditioning regimen in all cases. All patients received mycophenolate mofetile, cyclosporin A and short-term methotrexate with CD25 antibody as the graft-versus-host disease (GVHD) prophylaxis. The results showed that rapid engraftment was observed in all cases who presented full donor chimerism at 28 days post transplantation by STR-PCR. The median time of neutrophil recovery > 0.5 x 10(9)/L, platelet recovery > 20 x 10(9)/L was 13, 17.5 days respectively after transplantation. The incidence of acute GVHD was 3 cases (one case with grade I was recovered from GVHD by himself, one case with grade III was cured, one case with grade VI was died). It is concluded that above-mentioned preconditioning regimen and GVHD prophylaxis are effective approaches for unrelated donor peripheral blood stem cell transplantation in hematopoietic malignancies.

To investigate the changes of donor's peripheral blood immunocytes after mobilization with medium-dose recombinant human granulocyte-colony stimulating factor (rhG-CSF), the amounts of immunocytes in peripheral blood cells and the immunocyte components of donor peripheral blood mononuclear cells (PBMNC) in 12 healthy donors were detected by flow cytometry before and after mobilization with rhG-CSF 10 microg/(kg.day). The results showed that the median amounts of peripheral blood leukocytes before mobilization was 6.25 (4.7-7.8) x 10(9)/L, for lymphocytes it was 2.07 (1.63-3.10) x 10(9)/L, and for monocytes it was 0.163 (0.078-0.414) x 10(9)/L. In the fifth day after mobilization, the median amounts of peripheral blood leukocytes was 37.47 (24-72.57) x 10(9)/L, and for lymphocytes it was 3.22 (1.46-5.31) x 10(9)/L, and for monocytes, it was 1.2 (0.706-3.627) x 10(9)/L. The average amount of leukocytes after mobilization was 6.26 +/- 2.14 multiple of that before mobilization (P < 0.01), and the median amounts of lymphocytes after mobilization was 1.45 +/- 0.76 multiple of that before mobilization (P < 0.05), and the amount of monocytes after mobilization was 7.48 +/- 4.41 multiple of that before mobilization (P < 0.01). The median percentage of CD3(+) T lymphocytes before mobilization was 46.96% [(32.36-57.45)%], but 40.94% [(25.31-48.9)%] after mobilization. The ratio of CD4(+)/CD8(+) before mobilization was 1.27 +/- 0.46, while 1.36 +/- 0.51 after mobilization. The median percentage of CD4(+)CD8(+) T lymphocytes was 0.41% [(0.16-1.51)%], and 0.49% [(0.09-2.0)%] after mobilization. The median percentage of CD16(+)CD56(+) NK cells was 13.98% [(4.08-25.08)%] versus 16.65% [(12.06-33.05)%] after mobilization. The median percentage of CD3(+)CD16(+)CD56(+) NK-T cells was 2.75% [(0.37-6.38)%], but 3.13% [(0.46-5.95)%] after mobilization. The median percentage of CD20(+) B cells was 9.28% [(5.97-16.33)%], while 9.94% [(7.36-20.41)%] after mobilization. The median percentage of CD14(+) monocytes was 12.48% [(3.54-19.35)%] versus 29.52% [(16.51-36.76)%] after mobilization. The percentage of CD3(+) T lymphocytes, CD4(+)CD8(+) T lymphocytes, NK cells, NK-T cells and B lymphocytes in PBMNC did not change markedly before and after mobilization with middle-dose rhG-CSF. The ratio of CD4(+)/CD8(+) did not change significantly (P > 0.10). The percentages of CD14(+) monocytes in PBMNC after mobilization increased up to 2.87 +/- 1.51 higher than that before mobilization (P < 0.05). It is concluded that the changes of the CD14(+) monocytes after mobilization with rhG-CSF may be involved in graft rejection and graft versus host disease after allo-PBSCT.

In order to get clinical information about safety and feasibility of successively double autologous hemopoietic stem cell transplants (SD-AHSCT) in malignant hematological disease patients, the complications and hematological reconstitution after SD-AHSCT in 20 patients were analyzed retrospectively. 20 patients with hematologic malignancies received autologous peripheral blood stem/progenitor cell transplantation at the first transplant, and then were given autologous bone marrow transplantation as the second transplant at 4-10 months. The results showed that all the patients tolerated mobilization and collection of peripheral blood stem/progenitor cells as well as bone marrow collection. All the patients got enough hematological stem/progenitor cells for SD-AHSCT and achieved hematological reconstitution after SD-AHSCT. The speed of hematological reconstitution was positively correlated with the transfused quantity of hematological stem/progenitor cells (r = 0.968). The hematological reconstitution after the first autologous hemopoietic stem cell transplant (AHSCT) was earlier than that of the second (P < 0.05). There was no statistical difference between the first and the second AHSCT for the incidence of skin or mucous membrane bleeding (P > 0.05). No patients occurred massive hemorrhage during SD-AHSCT. The quantity of platelet transfusion in the second AHSCT was larger than that in the first AHSCT (P < 0.01). The incidence of oral ulcer in the first AHSCT was significantly higher than that in the second (P < 0.01). No statistical difference between the first and the second AHSCT was there in infectious sites, infectious pathogens and infection incidence (P > 0.10). All the complications were improved or cured, and no patients died of SD-AHSCT complications. In conclusion, SD-AHSCT is safe and feasible, and worthy to be further popularized.

To explore feasibility and efficacy of unrelated HLA matched donor marrow transplantation in treatment of myelodysplastic syndrome, one case (male, 31 years old) of myelodysplastic syndrome-refractory cytopenia with multilineage dysplasia (MDS-RCMD) has been received unrelated HLA-matched donor transplantation. Busulfan, cyclophosphamide, Ara-C, MeCCNU and antithymocyte globulin (ATG) were used for preparative regimen. Mycophenolate mofetile, cyclosporin A and short-term methotrexate were used for graft-versus-host disease prophylaxis. The results showed that neutrophil of > 0.5 x 10(9)/L, platelet of 58 x 10(9)/L and hemoglobin of 114 g/L were observed at 10, 20 days and 3 months respectively post transplantation. Disease-free survival without GVHD was 9 months. In conclusion, unrelated HLA matched donor marrow transplantation is an effective approach for treatment of patients with MDS.

To compare the clinical outcome of autologous peripheral blood stem cell transplantation (APBSCT) and autologous bone marrow transplantation (ABMT) in treatment of patients with acute leukemia in first remission, 41 patients received APBSCT, 17 patients received unpurged ABMT and 30 patients received purged ABMT. The results showed that hematopoietic recovery was significantly earlier after APBSCT than that after purged or unpurged ABMT. The 3-year disease-free survival (DFS), relapse rate (RR) and transplant-related mortality (TRM) for all patients of 3 groups were 51.7%, 41.7% and 6.8%, respectively. DFS and RR were significantly influenced by disease types (ALL or AML) and intervals between diagnosis and CR(1) or CR(1) and transplant. The main causes of transplant-related death were infection and hemorrhage. After APBSCT, DFS, RR and TRM were 48.4%, 43.9% and 4.9%, respectively, and did not differ significantly from those found in unpurged ABMT (47.1%, 45.6% and 11.8%) or purged ABMT (66.5%, 29.6% and 6.7%). It is concluded that the clinical outcome of APBSCT is similar to unpurged or purged ABMT but APBSCT allows faster recovery of hematopoiesis and needs less transfusion support.