This study was aimed to investigate various factors influencing long-term survival in adult AML patients with fusion gene aml1/eto positive. A single institutional retrospective study with long-term follow-up was performed to better define the prognostic factors for AML patients with aml1/eto positive. Newly diagnosed 89 adult AML patients with aml1/eto positive were followed up for 1 to 42 months (median 24 months) from January 2004 to July 2008. Univariate and multivariate analysis of potential factors influencing survival and prognosis were carried out by using Log-Rank and Cox regression method, including sex, age, initial WBC counts, extramedullary leukemic disease, central nervous system leukemia (CNSL), chromosome aberrations, immunophenotype, first induction regimen, chemotherapy course to complete remission (CR), time from induction therapy to CR, negative or positive rate of aml1/eto and allogeneic hematopoietic stem cell transplantation and so on. The results showed that the estimated 5-year overall survival (OS) and relapse-free survival (RFS) were (50.0 +/- 2.3)% and (47.0 +/- 1.9)% respectively in follow-up of 89 patients for 1 - 42 months (mean 24 months). Univariate analysis revealed that initial WBC counts, CNSL, chemotherapy course to CR, time from induction therapy to CR, persistent negative in remission and allogeneic hematopoietic stem cell transplantation were important prognostic factors for long-term surviva1. Multivariate study demonstrated that initial WBC counts, CNSL, CD56 positive, negative or positive rate of aml1/eto, time from induction therapy to CR, persistent negative result of RT-PCR assay in remission and allogeneic hematopoietic stem cell transplantation were all critical factors in relation to OS and RFS. It is concluded that Chinese adult AML patients with fusion gene aml1/eto positive have some different characteristics as compared with patients from other countries, a relatively poor outcome is observed in patients, HSCT should be recommended to adult AML patients.
Adolescent ; Adult ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; mortality ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Prognosis ; RUNX1 Translocation Partner 1 Protein ; Retrospective Studies ; Survival Analysis ; Young Adult

Natural killer (NK)/T-cell lymphomas represent a rare type of lymphoma derived from either activated NK cells or cytotoxic T cells. They are most commonly extranodal and tend to present as destructive lesions within the midline facial structures. Other than the nasal cavity and Para nasal sinuses, several other extra nodal sites of involvement have been reported, including the pharynx, gastrointestinal tract, and testis. Occasionally, pleural effusion has also been observed. Here, a case of lymphoma of NK/T-cell type presented as pleural effusion was reported. The patient was previously misdiagnosed as B cell non-Hodgkin's lymphoma by pathological and immunohistochemistry (IH) analysis for pleural membrane biopsy specimen. After the analysis of the pleural fluid cells by a combination of morphologic, immunophenotypic, cytogenetic and molecular (MICM) methods in Beijing Dao-Pei hospital, some lymphoblasts were found morphologically, which expressed cytoplasmic CD3 (cCD3) and CD56 by flow cytometry analysis and had a clonal T-cell receptor gamma (TCR-gamma) gene rearrangement by molecular analysis, so that the diagnosis was finally corrected as NK/T-cell lymphoma and an allogeneic stem cell transplantation was successfully performed. In conclusion, this unusual case highlights the significance of MICM combined techniques for the diagnosis of lymphoma, as well as an unusual presentation of a rare disease and the successful treatment.
Cytological Techniques ; Humans ; Lymphoma, Extranodal NK-T-Cell ; complications ; diagnosis ; Male ; Middle Aged ; Natural Killer T-Cells ; Pleural Effusion ; diagnosis ; etiology

OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley

Objective To explore the effeteness of autologous transplantation of bone marrow-derived endothelial progenitor cells in promoting the neovascularization and improving the neurological functional recovery in atherosclerotic rats with acute cerebral infarction. Methods Male Sprague-Dawley rat models of atherosclerosis were established by fat-rich diet feeding. Endothelial progenitor cells (EPCs) were obtained from bone marrow of all rats; the cells were cultured in vitro in Ml99 with VEGF, bFGF and EGF in it Assays were used to detect the expression of FLK-1 and CD34. on the 7th d, middle cerebral artery occlusion (MCAO) rat models were established by the method of thread thrombus. Three h after MCAO, all of the animals were randomized into experimental group (the autologous endothelial progenitor cells labeled with BrdU were injected into the carotid vein) and control group (same volume of PBS were injected into the carotid vein). Behavioral tests (modified neurological severity scale, mNSS) were performed 6 h and 1, 3, 7, 10 and 14 d after MCAO. Besides, immunohistochemical examinations were employed to observe the distribution of EPCs (labeled by BrdU) in the brain tissue and to measure the microvessel density. Results EPCs from bone marrow were isolated, induced and cultured successfully in vitro, which positively stained for FLK-1 by immunocytochemistry and partly positively expressed CD34 by immunofluorescence. The cells of FITC labeled UEA adsorption and DiL-acLDL internalization were positive under fluorescence confocal microscopy. These cells possessed robust proliferative potential and their number reached 5×106. On the 14th d, the neurological function recovery in the experimental group (mNSS scores: 6.13±0.30) was significantly improved as compared with that in the control group (mNSS scores: 8.50±0.46, P<0.05). On the 28th, some positive EPCs stained by BrdU were found in the experimental group and the numbers of blood vessels in the experimental group (16.87±5.52) were obviously larger than those in the control group (12.76±4.94,P<0.05). Conclusion EPCs from bone marrow can be autologously transplanted into the brain tissue of cerebral ischemia and survive. Cerebral ischemia can improve neurological functional recovery after MCAO, which were probably by promoting the neovascularization.

OBJECTIVE: To study DNA quantification and STR typing of samples pre-treated with pyramidon. METHODS: The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. RESULTS: In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. CONCLUSION Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.
Aminopyrine/pharmacology* ; Blood Stains ; DNA/isolation & purification* ; DNA Fingerprinting ; Forensic Medicine ; Humans ; Polymerase Chain Reaction ; Reproducibility of Results ; Specimen Handling

Three sesquiterpenoids and nine iridoids were isolated from the roots and rhizomes of Valeriana jatamansi by various chromatographic methods. Their structures were identified by physicochemical properties, NMR and MS data. Among them, valeriananoid G (1) was a new patchoulol-type sesquiterpenoid, and compound 3 was isolated from the genus Valeriana for the first time. Compounds 3 and 10 exhibited significant inhibitory effects on nitric oxide production induced by lipopolysaccharide in RAW 264.7 macrophages, with IC₅₀ values of 19.00 and 3.66 μmol·L^-1, respectively. In addition, compounds 4, 6 and 12 showed anti-influenza virus activity with IC₅₀ values of 51.75, 51.40 and 102.08 μmol·L^-1, respectively.

This study was purposed to analyze the characteristics of morphology, immunology, cytogenetic and molecular biology of leukemia cells in 12 AML patients with Ph(+) and their correlation with survival of patients. 12 patients with Ph(+) AML were diagnosed according to diagnostic criteria of WHO and existence of t(9;22) (q34;q11) or t(9;22) abnormality, meanwhile no evidence of CML chronic phase was observed. The results showed that 8 out of 12 cases were confirmedly diagnosed to be AML by morphologic and immunophenotypic examination, 4 cases were diagnosed as myeloid and B lymphocytic mixed acute leukemia. The Ph chromosome was detected in 10 cases by chromosome analysis at the first time of diagnosis, and some of the cases had coexistence of complex chromosome and/or normal karyotype. BCR-ABL transcript was detected in all 12 cases, including 7 cases with b3a2, 1 case with b2a2, 1 case with b2a2 variants, 2 cases with e1a2 and 1 case with e18a2. The 12 cases all got complete remission after chemotherapy and/or gleevec treatment, out of them 3 cases received chemotherapy and gleevec treatment, but 2 cases died; 9 cases received allogeneic hematopoietic stem-cell transplantation (allo-HSCT), 1 case died from relapse, among them 1 case died from transplant complications. The median survival was 24 (8 - 80) months, the overall survival of 3 years was (51.4 ± 17.7)%. It is concluded that the Ph(+) AML is a acute myelogenous leukemia with poor prognosis, but long-term survival may be achieved with HSCT as quick as after complete remission from gleevec and chemotherapy treatment. Meanwhile, the detection of BCR-ABL gene and it variants may be give more opportunity for diagnose and treatment, which can be used as routine screening for newly diagnosed leukemia.
Adult ; Child ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; Prognosis

OBJECTIVETo study the correlation between polyoma virus load and hemorrhagic cystitis after allogeneic stem cells transplantation for prevention of hemorrhagic cystitis.

METHODSBlood and urine specimens were collected from 40 healthy persons, 40 patient with stem cells transplantation and 20 cases complicated with hemorrhagic cystitis for determination of VP1 gene of polyomaviruses BK virus (BKV)/Jamestown Canyon virus (JCV) and simian virus 40 (SV40) by polymerase chain reaction (PCR) and EvaGreen stain fluorescence quantitative assay.

RESULTSIn the peripheral blood, all genes of BKV/JCV and SV40 were negative, while BKV gene in urine and blood from healthy persons and patient with stem cells transplantation was 15% (6/40) and 100% (40/40), respectively. The gene of JCV was positive in 10% (4/40) and 12% (5/40), the gene of SV40 was negative.

CONCLUSIONGenes of BKV and JCV was detectable in urine specimens of healthy persons and there was a correlation between the load of polyomavirus and incidence of hemorrhagic cystitis.

Capsid Proteins ; genetics ; Cystitis ; diagnosis ; etiology ; virology ; DNA, Viral ; blood ; genetics ; urine ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Hemorrhage ; diagnosis ; etiology ; virology ; Humans ; Polymerase Chain Reaction ; methods ; Polyomavirus ; genetics ; growth & development ; Polyomavirus Infections ; complications ; virology ; Viral Load

OBJECTIVETo study the effect of growth factor-primed donor hematopoietic stem cells infusion (GPBSCI) as an early adoptive immunotherapy.

METHODSTwelve patients with high-risk leukemia received prophylactic GPBSCI after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Out of the 12 patients, two were Ph(+) ALL in CR(1), one ALL in CR(2), one refractory ALL, three AML (one in relapse, one refractory disease, one in CR(3)), four CML in advanced stage and one myelodysplastic syndrome-refractory anemia with excess blast (MDS-RAEB).

RESULTSSixteen infusions were performed in the 12 patients, including 5 infusions were performed within +90 days post-SCT. The median mononuclear cells (MNC) and CD3(+) cells infused for GPBSCI before +90 d were 1.00 (0.95 - 1.24) x 10(8)/kg and 0.53 (0.39 - 0.63) x 10(8)/kg, and after +90 d were 2.27 (1.00 - 4.30) x 10(8)/kg and 1.15 (0.55 - 2.10) x 10(8)/kg, respectively. Four patients developed grade I - II acute GVHD, and one grade III acute GVHD. Seven patients developed chronic GVHD, of which four cases were extensive. Two patients had no transfusion related GVHD. No transfusion related pancytopenia was observed. Ten patients survived disease-freely at 563 (415 - 728) days of follow-up. Two patients died of leukemia relapse after GPBSCI.

CONCLUSIONAllo-HSCT with prophylactic GPBSCI could maximize graft-versus-leukemia effect with few fatal complications and might be a potentially curative strategy for hematological malignancy patients with high risk of relapse.

Adolescent ; Adult ; Blood Component Transfusion ; methods ; Child ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunotherapy, Adoptive ; Leukemia ; prevention & control ; surgery ; Male ; Middle Aged ; Postoperative Care ; Secondary Prevention ; Tissue Donors ; Transplantation, Homologous ; Treatment Outcome

The aim of study was to investigate the pharmacokinetics and distribution of antithymocyte globulin (ATG) in recipients of partially HLA-matched hematopoietic stem cell transplantation. Fifteen patients with hematological disorders were received hematopoietic stem cell transplantation from partially HLA-matched related donor between October 2003 and October 2004 in the Institute of Hematology and People Hospital, Peking University. All patients including 5 cases of AML, 6 cases of CML, 3 cases of ALL, 1 case of AA were consecutively enrolled in the present study after providing written informed consent. Antithymocyte globulin was administered before allogeneic hematopoietic stem cell transplantation at a dose of 2.5 mg/kg daily for 4 consecutive days (total dose of 10 mg/kg) in the conditioning regimen. The concentration of rabbit ATG in the serum of 15 patients was measured using a new enzyme-linked immunoabsorbent assay (ELISA) for the Fc portion of rabbit IgG. The results showed that the washout phase of ATG elimination was analyzed over 0 - 120 days, results were well-fitted by a single exponential decay giving a mean elimination half-life (t(1/2) beta) of 29.67 +/- 2.60 days. A mean value for the apparent volume of distribution of ATG (V) obtained by analysis of data was 0.12 +/- 0.02 L/kg body weight. The serum concentration of ATG increased up to 44.8% at 5 day before transplantation, peak concentration of ATG was 136.0 +/- 10.3 mg/L, its concentration slowly descend at 0 day, fall up to 7.1 +/- 0.06 microg/ml at 90 day after dosing; t(max) 4.8 +/- 0.7 days; According to AIC (Akaike's information criterion), two compartment model of ATG was estimated. It is concluded that the conditioning regimen containing the dosage of 10 mg/kg of ATG is effective and safely in recipients of partially HLA-matched hematopoietic stem cell transplantation. There is no racial difference in the pharmacokinetics of ATG.
Adolescent ; Adult ; Antilymphocyte Serum ; metabolism ; Child ; Female ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; Humans ; Male ; Middle Aged ; Tissue Distribution