Humans ; Male ; Middle Aged ; Pneumoconiosis ; diagnosis

A procedure for the preparation of PCR template from Saccharomyces cerevisiae using boiling method is described,and arg-13 gene from low copy ARSCEN plasmid and ymc1 gene from genomic DNA are amplified with high efficiency respectively.

Biopsy ; Biopsy, Needle ; Bone Marrow ; pathology ; Bone Marrow Examination ; methods ; Bone Marrow Neoplasms ; pathology ; secondary ; Breast Neoplasms ; pathology ; Cytological Techniques ; Female ; Humans ; Lung Neoplasms ; pathology ; Male ; Prostatic Neoplasms ; pathology ; Retrospective Studies ; Stomach Neoplasms ; pathology

OBJECTIVETo detect p15(INK4B) methylation levels and the kinetics of the methylation status before and after decitabine to explore its influences on prognosis and response to decitabine in myelodysplastic syndromes (MDS) patients.

METHODSWe examined 261 MDS patients (143 male and 118 female) with the median age of 52 years (32-78). Of them, 172 cases were low-risk group (low-risk 104 cases, intermediate-1 68 cases), 89 cases high-risk group (intermediate-2 52 cases, high risk 37 cases). Collections of bone marrow mononuclear cells of MDS patients and extracted the genomic DNA, the methylation status of p15(INK4B) was detected by methylation-specific PCR (MSP) method. Survival analysis was conducted according to the level of p15(INK4B) methylation in the cohort of patients. The kinetics of the methylation levels of p15(INK4B) in 58 MDS patients before and after 2 courses of decitabine have been assessed with the method of MSP.

RESULTSThe methylation level of p15(INK4B) in low-risk group patients was significantly lower than that in high-risk group (117.22 vs 157.63, P<0.05 ). The expected 2-year survival rate of p15(INK4B) methylation positive patients was lower than that of negative ones (91.8% vs 69.8%, P<0.05); the expected 2-year survival rate of p15(INK4B) methylation positive patients was shorter than that of negative ones in low-risk group（78.2% vs 92.0%, P<0.05）, meanwhile there was no significant difference in terms of expected 2-year survival rate and median expected survival between p15(INK4B) methylation positive and negative patients in high-risk group [35.6% vs 38.5%, (17.0±9.3) month vs (18.0±5.7) month, P>0.05]. Multivariate analysis showed p15(INK4B) methylation degree was an independent prognostic factor for overall survival. No statistical difference of overall response (OR) rates were found between p15(INK4B) methylation positive patients and negative patients before decitabine（65.9% vs 76.5%, P>0.05）, and complete remission (CR) rates between these two groups also showed no statistical difference（22.0% vs 29.4%, P>0.05）. p15(INK4B) methylation levels had no obvious change before and after treatment in decitabine responders（P>0.05）.

CONCLUSIONThe survival of newly diagnosed MDS patients with positive p15(INK4B) methylation was comparatively shorter, but p15(INK4B) methylation had no influence on response to decitabine.

Adult ; Aged ; Azacitidine ; analogs & derivatives ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; drug therapy ; genetics ; Prognosis ; Survival Rate ; Treatment Outcome

OBJECTIVETo explore the clinical value of urine N-telopeptides of type I collagen (uNTX) and serum bone specific alkaline phosphatase (sBAP) in myeloma bone disease, and to understand the role of bisphosphonates therapy for multiple myeloma(MM) osteolytic bone lesion.

METHODSThirty-three MM cases were treated with bisphosphonates combined with chemotherapy (considered as treatment group), and 20 untreated MM cases with chemotherapy alone considered as control group. uNTX was detected by ELISA, and sBAP by chemiluminescence analysis.

RESULTS(1) There was no significant differences in uNTX between treatment \[(173.74 ± 14.55) µg/L\] and control groups \[(129.79 ± 12.13) µg/L\] before bisphosphonates treatment (P > 0.05). After six-month treatment, there was significant differences between two groups \[(85.71 ± 8.23) µg/L and (121.59 ± 12.43) µg/L, respectively\] (P < 0.05); Meanwhile, there were significant differences in uNTX between before and after three-month treatment (P = 0.045) and between before and after six-month treatment (P < 0.01) in treatment group. (2) There was no significant differences in sBAP concentration between treatment and control groups \[(4.78 ± 0.55) µg/L and (8.42 ± 1.32) µg/L, respectively\] before treatment (P > 0.05). After six-month treatment, there were significant differences between them \[(16.01 ± 0.52) µg/L and (9.62 ± 1.29) µg/L, respectively\] (P < 0.01). Meanwhile, in treatment group, there was no significant differences between before and after three-month treatment (P > 0.05), but being significant difference between before and after six-month treatment (P < 0.01).

CONCLUSIONuNTX, sBAP are important early sensitive index to measure the osteolytic bone lesion in MM patients. Bisphosphonates can significantly improve the osteopathy in MM cases.

Adult ; Aged ; Alkaline Phosphatase ; blood ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Density ; Bone and Bones ; metabolism ; Collagen Type I ; urine ; Diphosphonates ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; metabolism

OBJECTIVETo study the methylation status of p73 gene promoter in patients with myelodysplastic syndrome (MDS) and explore its significance with clinical prognosis.

METHODSMethylation of p73 promoter was detected in bone marrow cells from 135 MDS patients and 13 healthy controls by methylation-specific PCR (MSP). The results of MSP were confirmed by bisulfite sequencing. The expression of p73 mRNA was detected by real-time quantitative PCR. Primary bone marrow cells from MDS patients were treated with decitabine, the changes of p73 methylation status and p73 mRNA expression were measured. The role of p73 methylation in the prognosis of MDS and the correlated clinical data were explored.

RESULTSp73 hypermethylation was present in 37.04% of MDS cases and patients with high risk MDS (RAEB-1 and RAEB-2) exhibited a significantly higher frequency of p73 methylation than that of low risk MDS (58.8% vs 29.7%, P = 0.002). The expression of p73 mRNA in the methylated group was decreased compared to that of the unmethylated group (P = 0.032). Decitabine treatment decreased the level of p73 methylation and increased the level of p73 transcripts. Patients with p73 methylation progressed rapidly to AML (P < 0.001) and had shorter survival (P = 0.002) than those who did not have p73 methylation. In the multivariate Cox regression model, BM blast and p73 methylation status emerged as independent prognostic factor for overall survival and leukemia free survival.

CONCLUSIONp73 gene methylation is common in patients with MDS and may indicate poor prognosis. p73 may be a therapeutic target in MDS.

Aged ; Case-Control Studies ; DNA Methylation ; DNA-Binding Proteins ; genetics ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Nuclear Proteins ; genetics ; Prognosis ; Promoter Regions, Genetic ; Tumor Protein p73 ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo investigate the effects of ulinastatin (Uti) and low-molecular-weight heparin (Lmwh) on coagulation function and deep vein thrombosis (DVT) in patients undergoing hip joint replacement.

METHODSFrom March to December 2010 150 ASAI-II patients with average age of 72.5 (65 - 85) years undergoing hip joint replacement were randomly divided into 3 groups (n = 50 each): normal saline (NS) control group (Group C), Uti group (Group U) and Lmwh group (Group L). Group U received intravenous infusion of ulinastatin (10 000 U/kg) at preoperative, perioperative and after operation 1, 2 and 3 d, respectively. Group C received the same volume of NS instead of Uti. Group L were injected Lmwh subcutaneously (3200 U/d) at preoperative, after operation 1, 2 and 3 d. Blood samples were taken before operation (T(0)), at the end of surgery (T(1)), 1 d (T(2)), 2 d (T(3)) and 3 d (T(4)) after operation for determination the values of R, K, α angle, MA and CI, using thromboelastography, and the DVT were also examined through color Doppler ultrasonography at 3 d after operation.

RESULTSCompared with T(0), R, K were shorter, α angle, MA and CI were larger in group C, the values at T(2) were up to the peak then declined at T(4). Compared with group C, the value of R, K were larger, the value of α angle, MA and CI were shorter in group U and group L. The DVT checked by ultrasonography were found in 20 cases in group C, 1 case in group U, and zero case in group L. The differences were no statistically significant between group U and group L.

CONCLUSIONIntravenous infusion of Uti during the period of operation can correct the hypercoagulability of blood and decrease the incidence of DVT after operation.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; Blood Coagulation ; Double-Blind Method ; Female ; Glycoproteins ; therapeutic use ; Heparin, Low-Molecular-Weight ; therapeutic use ; Humans ; Male ; Postoperative Complications ; prevention & control ; Venous Thrombosis ; etiology ; prevention & control

OBJECTIVETo explore the effect of compound qizhu granule (CQG) on cellular immunity of chronic hepatitis B (CHB) patients.

METHODSTotally 103 CHB patients treated with lamivudin (LAM) for 6 months, who had partial virological response (HBeAg positive) were randomly assigned to two groups, 50 in the treatment group and 53 in the control group. All patients took LAM 100 mg (once a day) plus ADV 10 mg (once a day). Patients in the treatment group additionally took CQG, one dose per day. After one-year treatment hepatitis B virus (HBV) DNA negative rates, HBeAg seroconversion, levels of HBV specific cytotoxic T lymphocyte (CTL), non-specific CTL and natural killing (NK) cells were compared between the two groups.

RESULTSAfter 1-year treatment, HBV DNA negative rate of the treatment group was 88: 0% in 44 cases, slightly higher than that of the control group (41 cases, 77.4%), but with no statistical difference (P >0.05). HBeAg seroconversion of the treatment group was 32.0% in 16 cases, higher than that of the control group (8 cases, 15.1%), with statistical difference (P <0.05). Levels of HBV specific CTL (0.79%±0. 07%), non-specific CTL (19.4%±1.8%) and NK cells (14. 1%± 1.5%) of the treatment group were higher than those of the control group (0.58% ± 0.08%, 17.5% ± 1.7%, and 11.1%±1.5%, respectively; allP <0.01).

CONCLUSIONTreating CHB patients with partial virological response by ADV plus CQG could improve specific and non-specific cellular immunity, thereby elevating HBeAg seroconversion rate.

Drugs, Chinese Herbal ; therapeutic use ; Hepatitis B e Antigens ; immunology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Immunity, Cellular ; immunology ; T-Lymphocytes, Cytotoxic ; drug effects

OBJECTIVETo investigate phenotype, cell differentiation and cytogenetic properties of bone marrow (BM) mesenchymal stem cells (MSC) separated from the myelodysplastic syndrome (MDS) patients. And to analyze cytogenetic aberration of MSC derived from MDS (MDS-MSC) and its mechanism in pathogenesis of MDS.

METHODSAdherent MSC from both myelodysplastic (n = 22) and normal (n = 7) marrow were obtained by a stromal culture procedure. Morphological features were observed by optical microscope. The cell-surface antigens were performed by flow cytometer(FCM). Adipogenic and osteogenic differentiation potential of MSC were identified under specific induction conditions. Standard cytogenetic analysis of both hematopoietic cells and MSC were performed by trypsin-Giemsa (GTG) banding. The karyotype analysis DNA content was determined by FCM to verify the results.

RESULTSThe morphology of MDS-MSC was typical slender spindle-shaped cells, MSC obtained from MDS patients had a MSC immunophenotype, lacked the expression of hematopoietic antigens-CD34, CD45 and expressed MSC markers, such as CD73, CD90, and CD105. MDS-MSC layers showed the capability to differentiate towards adipocytes, chondrocytes and osteoblasts. Cytogenetic aberrations were observed in MSC from 14 (64%) MDS patients, usually involve the loss of chromosomal material (92%), and the clonal loss (7 cases, 50%). Two cases of structural aberrations were also detected. Abnormal karyotypes in MSC were still more frequently identified in abnormal hematopoietic cells group (12 out of 13, 92% vs 3 out of 9, 33%, P < 0.05). There were not exactly the same type of chromosomal aberrations between hematopoietic cells and MSC, but different type of the aberrations in the same chromosome were involved.

CONCLUSIONMDS-MSC retains the phenotyping characteristics and differentiated function of normal MSC, but has different type of chromosomal abnormalities. A high proportion of loss of chromosomal may be a marker of chromosomal instability of MDS-MSC. Detection of abnormalities in MDS-MSC suggests enhanced genetic susceptibility of these cells in MDS. This may indicate potential involvement of MSC in the pathophysiology of MDS.

Adult ; Aged ; Aged, 80 and over ; Bone Marrow Cells ; cytology ; Case-Control Studies ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Karyotyping ; Male ; Mesenchymal Stromal Cells ; cytology ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; immunology ; Phenotype ; Young Adult

This study was aimed to investigate the influence of timing using G-CSF after chemotherapy on graft yield of mobilized peripheral blood stem cells for autoPBSCT. 39 patients with lymphoma or multiple myeloma (MM) received the same chemotherapy mobilization regimen, including CTX 400 mg/m² d1; VLB 2 mg/m(2) d1; Ara-C 60 mg/m ²× d1-5; VP-16 60 mg/m² × d1-5; and prednisone 40 mg/m² × d1-5. The historical control group (12 cases) received G-CSF subcutaneously (filgrastim) at the first restoration after the initial nadir of the peripheral WBC count. The experimental group (27 cases) received G-CSF during the steady rise of the WBC count (end of fluctuating after initial nadir). G-CSF was given in a single daily subcutaneous dose of 5 µg/kg until the final PBSC apheresis. When the peripheral WBC and mononuclear cell (MNC) counts reached 10 × 10⁹/L and 1.0 × 10⁹/L respectively, leukapheresis was carried out using the COBE Spectra blood cell separator. The results indicated that despite there was comparable treatment with alkylating agents between 2 groups, a significantly increased yield of CD34 positive cells was observed in the experimental group (26.4 × 10⁶/kg), as compared to the historical control group (3.1 × 10⁶/kg) (p = 0.0031). It is concluded that the appropriate timing for the use G-CSF mobilization after chemotherapy is important to increase the CD34(+) cell yield in auto-graft.
Adult ; Antigens, CD34 ; Antineoplastic Combined Chemotherapy Protocols ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Lymphoma ; therapy ; Male ; Middle Aged ; Multiple Myeloma ; therapy ; Transplantation, Autologous ; Young Adult