Humans ; Myeloproliferative Disorders ; genetics ; Receptor, Fibroblast Growth Factor, Type 1 ; genetics

Hepatitis B virus ; drug effects ; physiology ; Hodgkin Disease ; drug therapy ; Humans ; Immunoconjugates ; therapeutic use ; Virus Activation ; drug effects

OBJECTIVETo identify methylation profiles in myelodysplastic syndrome (MDS) and to provide the biomarkers for the early diagnosis and differential diagnosis of MDS.

METHODSGenes were screened for hypermethylation by genome-wide DNA methylation profiles. Transcription down-regulation was determined with a gene expression microarray. Methylation-specific, real-time, and bisulfite-sequencing PCR cloning and sequencing were performed to validate selected genes in MDS cases and non-malignant hematologic diseases (controls). Diagnostic test, such as sensitivity and specificity, was used to evaluate the value of methylation patterns.

RESULTSA draft of methylation patterns was established and refined to 6 genes after validation in 211 patients and 60 controls. The hypermethylated genes were ABAT (97%), DAPP1 (98%), FADD (89%), LRRFIP1 (96%), PLBD1 (89%), and SMPD3 (85%). A combination of 5 or more than 5 genes showed a specificity of 95% and sensitivity of 91.4% for the diagnosis of MDS. The accuracy of diagnosis was 92.3%.

CONCLUSIONWe demonstrated here that the ABAT, DAPP1, FADD, LRRFIP1, PLBD1 and SMPD3 genes are hypermethylated and downregulated in MDS. The six genes could be the markers of the methylation patterns in MDS, as a noninvasive approach for the diagnosis of MDS.

DNA Methylation ; Down-Regulation ; Gene Expression ; Genome, Human ; Humans ; Myelodysplastic Syndromes ; diagnosis ; genetics

OBJECTIVETo analyze the detection rate and gene distribution characteristic of deletional α-thalassemia in Guangxi area, and to provide theoretic basis for thalassemia gene diagnosis and genetic counseling.

METHODSThe regular gene diagnosis of 3 types of α-thal (-- (SEA),- α(3.7),- α(4.2)) was performed by gap-PCR, multiple ligation probe and gene sequencing for globin α or β were used to detect those samples whose genotype and phenotype were not consistent. And the distribution characteristic of α-thalassemia gene in Guangxi area was then analyzed.

RESULTSOut of 51 191 suspected thalassemia patients, there were 19 853 cases of deletional a-thalassemia, accounted for 39.9% in total positive rate, including 19 780 cases of regular types(--(SEA), - α(3.7), - α(4.2)), 61 cases of Thailand-type deletion, 9 cases of triplet type (Hong Kong) (ααα(HK)), 1 case of 21.9 kb deletion type and 2 cases of 809 bp deletion type.

CONCLUSIONTypes of deletional a-thalassemia were complex and accounted for large proportion in Guangxi area. Special gene diagnoses were needed for those couples whose genotype and phenotype were not consistent, in order to provide reliable basis for genetic counseling and prenatal diagnosis.

China ; Genotype ; Humans ; Phenotype ; Polymerase Chain Reaction ; Sequence Deletion ; alpha-Thalassemia ; genetics

OBJECTIVETo investigate the effect of the lentiviral vector mediated CXCR4 overexpressed mesenchymal stem cell (MSCs) on graft-versus-host disease (GVHD).

METHODSLentiviral vector containing CXCR4 was constructed. CXCR4 overexpressed MSC by lentiviral vector mediated were assessed. A major histocompatibility complex (MHC)-mismatched mouse model of bone marrow transplantation (BMT) from C57BL/6 donors to BALB/c recipients was constructed. Mice were divided into five groups: total body irradiation (TBI) group, mice received irradiation only; BMT group, mice were transplanted with bone marrow (BM) after TBI; GVHD group, mice were transplanted with BM and splencytes after TBI; CXCR4-MSC group, mice were transplanted with CXCR4-MSC, BM and splencytes after TBI; EGFP-MSC group, mice were transplanted with EGFP-MSC, BM and splencytes after TBI. The survival, body weight and clinical score of GVHD in transplanted mice were monitored. Liver, intestine and skin from mice in each group were obtained for histological examination. Plasma concentrations of inflammation factors such as interleukin (IL)-2, IFN-γ and TNF-α were also determined using a cytometric bead array cytokine kit.

RESULTSAll mice in TBI group died within 14 days, while all of BMT group survived. The mean survival times for GVHD, EGFP-MSC and CXCR4-MSC groups were (17.0 ± 2.3) d, (21.7 ± 4.8) d and (30.1 ± 9.1) d, respectively. Treatment with CXCR4 over-expressing MSCs could decrease the mortality rate. All mice in each group developed clinical signs such as hunched posture, dull fur, diarrhea and weight loss. Meanwhile, histopathological findings in target organs were confirmed the presence of GVHD. While, clinical GVHD scores and histopathological scores in CXCR4-MSC group were significantly lower than that of GVHD group. Moreover, compared with control groups, the plasma IL-2, IFN-γ and TNF-α level in recipients infused with CXCR4-MSC were significantly decreased (P<0.05).

CONCLUSIONThe results revealed that CXCR4- transduced MSCs could effectively control the occurrence of mouse GVHD following allogeneic BM transplantation.

Animals ; Bone Marrow Transplantation ; Cytokines ; Genetic Vectors ; genetics ; Graft vs Host Disease ; Lentivirus ; genetics ; Mesenchymal Stromal Cells ; immunology ; Mice ; Receptors, CXCR4 ; genetics ; immunology ; Transplantation, Homologous

OBJECTIVETo investigate the effect and mechanism of high dose Vitamin B3 on granulopoiesis in normal rat.

METHODSTwenty one healthy SD rats were randomly divided into three groups: the Vitamin B3 group (Vit B3 500 mg·kg⁻¹·d⁻¹, × 7 d), the rhG-CSF group (rhG-CSF 25 μg·kg⁻¹·d⁻¹, × 7 d) and the normal saline group (2 ml/d, × 7 d). The peripheral blood cell counts were analyzed by automatic blood cell counter before (day 0) treatment, the third day (day 3) and the seventh day (day 7) after administration of drugs, respectively. The concentration of serum nicotinamide adenine dinucleotide (NAD⁺) level was measured by enzymatic cycling assay before and after drugs treatment. The expressions of G-CSF, G-CSFR, SIRT1, C/EBPα, C/EBPβ, C/EBPε and NAMPT mRNA were detected by reverse transcription real-time fluorescent quantitative PCR.

RESULTSThe neutrophil counts increased significantly after 7 days of Vitamin B3 and rhG-CSF treatment compared with that of control group [(1.64 ± 0.19) × 10⁹/L, (1.88 ± 0.37)× 10⁹/L vs (0.86 ± 0.18) × 10⁹/L, P<0.01]; the level of serum NAD⁺ increased significantly [(0.96 ± 0.08) nmol/L, (0.65 ± 0.12) nmol/L vs (0.36 ± 0.15) nmol/L, P<0.01]; the expression of G-CSF, G-CSFR, SIRT1, C/EBPα, C/EBPε and NAMPT mRNA in bone marrow mononuclear cells were increased significantly compared with that of control group (P<0.01).

CONCLUSIONHigh dose of Vitamin B3 may play an important role in increasing absolute neutrophil count in healthy rat under steady state, and the mechanism may be dependent on NAMPT-NAD⁺-SIRT1 signaling pathways.

Animals ; Bone Marrow Cells ; Granulocyte Colony-Stimulating Factor ; Leukocyte Count ; Neutrophils ; drug effects ; Niacinamide ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins

OBJECTIVETo investigate the proliferation inhibitory role and mechanism of PI3Kδ inhibitor CAL-101 on multiple myeloma (MM) cells, and to provide new therapeutic options for MM treatment.

METHODSMM cell lines U266 and RPMI8226 cells were treated with various concentrations of CAL-101. MTT assay and CalcuSyn software were performed to determine the inhibitory effect of CAL-101 and the synergistic effect with PCI- 32765, SAHA (suberoylanilide hydroxamic acid), BTZ (Bortezomib) on MM cells. The protein expression level of p-AKT, p-ERK, AKT, ERK and PI3Kδ processed by CAL-101 were analyzed by Western blot.

RESULTSCAL-101 at concentration of 15, 20, 25, 30 and 40 μmol/L could induce significant dose-dependent proliferation inhibition on U266 cells after treatment for 48 hours. The cell proliferation inhibition rates were (33.54 ± 1.23)%, (41.72 ± 1.78)%, (53.67 ± 2.01)%, (68.97 ± 2.11)% and (79.25 ± 1.92)%, respectively. Similar results were found in RPMI8226 cell line. Western blots showed high expression level of p-AKT, p-ERK, AKT, ERK and PI3Kδ in cell lines and MM primary cells. p-AKT and p-ERK protein expression levels were down-regulated significantly by CAL-101 treatment. Synergistic effect has been verified between CAL-101 and PCI-32765, SAHA and Bortezomib in U266 cell line, and PCI-32765, Bortezomib in RPMI8226 cell line with CI values less than 1.

CONCLUSIONCAL-101 could inhibit proliferation of MM cell lines. High levels of p-AKT, p-ERK, AKT, ERK and PI3Kδ protein expression were observed in both cell lines and primary cells. Down-regulation of p-AKT and p-ERK probably related with the mechanism of CAL-101 in MM cell proliferation inhibition. CAL-101 has significant synergistic effect with PCI-32765, SAHA and BTZ.

Boronic Acids ; Bortezomib ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Humans ; Multiple Myeloma ; pathology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Protein Kinase Inhibitors ; pharmacology ; Purines ; pharmacology ; Pyrazines ; Pyrazoles ; Pyrimidines ; Quinazolinones ; pharmacology

OBJECTIVETo investigate the clinical and hematological features and prognostic factors of primary myelofibrosis (PMF) patients in Shanghai and surrounding area of Shanghai, China.

METHODSThe clinical manifestations, laboratory parameters and survival were retrospectively analyzed in 75 PMF patients diagnosed from Jan, 1996 to Dec, 2013 in our hospital and were compared with those of Chinese subjects from Tianjin and foreign Caucasian patients, respectively. Comparison of categorical variables was performed by χ² test. Survival was estimated using the Kaplan- Meier method. Log- rank test was used to compare survival date. A Cox model was used for multivariate analyses.

RESULTSThe median age of the 75 patients was 56(19-81) years old. There were 51(68%) patients with HGB less than 100 g/L. The median value of HGB was 83 g/L. Similar with those from Tianjin, the patients in our study were significantly younger with higher proportion of severe anemia and lower platelet counts when compared with foreign Caucasian patients. Using IPSS and dynamic international prognostic scoring system (DIPSS) model, the survival curves of intermediate- 1, intermediate- 2 and high risk groups were significantly different. In univariate analyses, variables significantly correlated with poor prognosis were systemic symptoms, HGB<100 g/L, HGB<80 g/L, PLT ≤ 100 × 10⁹/L, WBC<10 × 10⁹/L and blood blasts ≥ 0.01. Multivariate analysis identified IPSS and HGB < 80 g/L as independent risk factors for survival.

CONCLUSIONChinese PMF patients may have characteristics of younger age at onset with more severe anemia. However, IPSS and DIPSS model are still suitable for Chinese patients to predict survival. To increase the weight of anemia severity may provide a better prognostic stratification for Chinese patients with PMF.

Adult ; Aged ; Aged, 80 and over ; China ; Humans ; Middle Aged ; Primary Myelofibrosis ; diagnosis ; Prognosis ; Retrospective Studies ; Risk Factors ; Young Adult

OBJECTIVETo analyze the clinical features, prognostic factors, diagnostic methods and treatment outcomes of primary breast lymphoma (PBL).

METHODSThe clinical data of 14 patients diagnosed with PBL between 2000.1 and 2013.6 were analyzed retrospectively.

RESULTSThe 14 patients were diagnosed with PBL, which comprised 0.24% and 0.54% of all breast malignancies and lymphoma, respectively. The median age was 43(20-77) years. All but one was female. The median course before diagnosis was 1(0.17-12) month. There were 9 patients with international prognostic index (IPI) 0 and 5 with IPI 1. The most common histological subtypes were diffuse large B cell lymphoma (DLBCL) with total 11 cases (78.6%), there was 1 case (7.1%) in each of extranodal margin zone lymphoma, peripheral T cell lymphoma(PTCL) and small lymphocytic lymphoma (SLL), respectively. Patients treated with radical operation versus local mass removing or needle biopsy were 6(42.9%) and 8(57.1%), respectively, there were 2 relapses in each group. Patients treated with or without rituxinmab combined with chemotherapy were 6(42.9%) and 7(50.0%), respectively, there were 3 and 1 relapses in each group, respectively. Three (21.4%) patients received intrathecal injection (IT). There were 3(21.4%) cases of central nervous system (CNS) relapse, who were not received IT. After the median follow-up of 45.2 (10.7-116.1) months, two patients died of disease progression. The median overall survival did not reach and median progression free survival was 73 (11- 116) months.

CONCLUSIONThe most common histological subtype in patients with PBL was DLBCL, the role of rituxinmab in the treatment was not sure, CNS relapse should be monitored closely.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; Breast Neoplasms ; Disease-Free Survival ; Female ; Humans ; Lymphoma ; Middle Aged ; Neoplasm Recurrence, Local ; Retrospective Studies ; Young Adult

OBJECTIVETo report the diagnosis, differential diagnosis and treatment of three rare cases of primary bone marrow diffuse large B cell lymphoma (DLBCL), and to improve the recognition of this disease.

METHODSThe clinical characteristics, therapeutic course and the outcome of these patients were reviewed. Meanwhile, a series of examinations including morphology, flow cytometry, immunohistochemistry and molecular biology of bone marrow samples were also performed.

RESULTSThese three patients who were old at the onset age (56, 60 and 70 years old), primarily revealed as abnormal blood count and experienced an aggressive course of disease. Physical and imaging examination showed no enlargement of lymph node, liver and spleen, the patients were finally diagnosed as primary bone marrow DLBCL by bone marrow morphology, flow cytometry and immunohistochemistry analyses. They were treated with rituximab combined chemotherapy, which achieved a complete response, but still need longer follow-up to further evaluate their survival.

CONCLUSIONPrimary bone marrow DLBCL was encountered rarely in clinical practice, and this is the first report in China. Further investigation of pathogenesis and therapeutic strategies of this rare disease was warranted.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Marrow ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; Middle Aged