The study was purposed to investigate the significance of id4 gene promoter methylation in monitoring AML patients with complete remission (CR). Methylation specific-PCR (MS-PCR) were used to detect the status of promoter methylation of id4 gene in bone marrow samples from AML patients with CR who had accepted induction with DA or IA and 4 to 5 consolidation chemotherapy with Ara-C. The results showed that in the all 32 patients, 15 were found to show id4 promoter methylation and 7 out of the 15 were found relapsed or tendency to relapse in the following-up period. While all the 17 patients with id4 unmethylation were still in their CR status in the same period. In conclusion, id4 promoter methylation might be a predictor for relapse of AML patients with CR in certain degree.
DNA Methylation ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Neoplasm, Residual ; diagnosis ; genetics ; Promoter Regions, Genetic ; genetics

OBJECTIVEIt is necessary to compensate the system latencies in real-time tumor-tracking radiotherapy by prediction. However, due to the irregularities of respiratory motions, the results obtained with traditional methods were not acceptable. The purpose of this study is to evaluate the value of nonparametric regression model in respiratory motion prediction.

METHODSThe data of respiratory trajectory of 11 volunteers were obtained and predicted based on nonparametric regression method. The results were compared with those of autoregressive model and back propagation neural network. An improved method was proposed to deal with the abnormal state in respiration. We combined the prediction method with the tracking system to test its performance in practical application.

RESULTSThe results indicated that the proposed method could predict the motion accurately in real-time for different latencies. This method decreased the error of the abnormal state substantially and also allowed effective prediction of respiration motion when combined with the tracking system.

CONCLUSIONThe nonparametric regression model can predict the respiratory motion accurately in real-time and therefore meets the requirement of real-time tumor-tracking radiotherapy.

Forecasting ; Humans ; Models, Theoretical ; Movement ; Neoplasms ; radiotherapy ; Radiotherapy, Computer-Assisted ; methods ; Regression Analysis ; Respiration

The aim of this study was to investigate the difference of Id4 gene promoter methylation in patients with different subtypes of myelodysplastic syndromes (MDS). By using MS-PCR method, the methylation status of Id4 gene was detected in 50 patients with different subtypes of MDS. Id4 methylation was also detected in bone marrow samples from patients with iron deficiency anemia which served as control. The results showed that Id4 gene was unmethylated in all of bone marrow samples from controls. In various subtypes of MDS patients, the rate of Id4 gene methylation was different. No Id4 methylation was found in 6 cases of RA, 2 cases of RARS and 4 cases of MDS-U. Id4 methylations was found in 2 out of 18 patients with RCMD. Id4 methylation in 3 out of 12 patients with RAEBI and other 3 out 8 patients with RAEBII were found. In groups with blast ratio lower or higher than 5%, the incidence of Id4 gene methylation were 6.7% and 30% respectively, so that there was significant difference. In tentative conclusion, Id4 gene methylation possibly is found in MDS patients with higher ratio of blast cells.
Adult ; Aged ; DNA Methylation ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Male ; Middle Aged ; Myelodysplastic Syndromes ; classification ; genetics ; Promoter Regions, Genetic

Adoptive immunotherapy using allogeneic natural killer (NK) cells provides to be useful in recipients after allogeneic hematopoietic stem cell transplantation (Allo-HSCT), but its application has been limited by the inability to obtain sufficient numbers of pure NK cells. This study was aimed to optimize the expansion of high purity NK cells from human peripheral blood. First, the NK cells were isolated from PBMNC by using miniMACS (magnetic cell-selection) and NK Cell Isolation Kit II. Then the isolated cells were cultured in SCEM (Stemline Hematopoietic Stem Cell Expansion Medium, Sigma) supplemented with 10% human AB serum and different combinations of IL-2 and/or IL-12, IL-15 for 15 days. Cultures were fed with fresh media and cytokines every 3 days, and were evaluated for cell expansion, phenotype, and cytotoxicity at the end of the culture period. The results showed that in group IL2 + IL15 and IL2 + IL15 + IL12, cells were expanded 50.46 +/- 4.31 and 52.35 +/- 6.72-fold respectively, much higher than others (P<0.01), but no significant difference between themselves (P>0.05). And the purity of CD3(-)CD56(+) NK cells was over 94% in all groups except the control. The cytotoxicity of expanded NK cells cultured with cytokines was significantly higher than the starting population at different E:T ratio (P<0.01), although the cytotoxicity of IL2 + IL15 + IL12 group was slightly higher than that of IL2 + IL15 group, but no significant difference between themselves (P>0.05). It is concluded that high purity of NK cells can be efficiently expanded in culture with IL2 + IL15.
CD3 Complex ; analysis ; CD56 Antigen ; analysis ; Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Cell Separation ; Cells, Cultured ; Culture Media ; Humans ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Killer Cells, Natural ; cytology ; immunology ; Leukocytes, Mononuclear ; cytology

This study was purpose to investigate the difference of Id4 gene promoter methylation between healthy individuals and acute leukemia patients. MS-PCR methods were used to detect the status of Id4 gene methylation in healthy individuals and acute leukemia patients. The results showed that Id4 gene was unmethylated in bone marrow samples from healthy individuals. In new diagnosed AML and ALL patients, the rate of Id4 gene methylation was 84% and 86% respectively. Id4 gene methylation was found in all 8 cases of relapsed acute leukemias. In 14 ALL-CR patients with Id4 gene methylation, 8 patients relapsed within 12 months, while in 9 ALL-CR patients with Id4 gene unmethylation only 1 patient relapsed within 12 months. In AML-CR patients, the 12-months relapse rate in patients with Id4 gene methylation was 62.5%, while it was 10% in Id4 gene unmethylation patients, there was significant difference between them. The rate of Id4 gene methylation in ALL-CR patients was 64.3%, while it was 28.6% in AML-CR patients, there was significant difference between them. In the all 39 new diagnosed AL patients, Id4 gene methylation could be detected in 33 patients. In all 8 relapsed AL patients Id4 gene methylation was found, out of 58 AL-CR cases, 24 patients was found with Id4 gene methylation. It is concluded that as compared with healthy individuals, Id4 gene in acute leukemia patients was methylated in different degrees. The percentage of Id4 gene methylation in AL-CR group is lower than that in AL-non remission group. The change of Id4 gene methylation is thought to be associated with occurrence of acute leukemia.
Acute Disease ; Case-Control Studies ; DNA Methylation ; Female ; Humans ; Inhibitor of Differentiation Proteins ; metabolism ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics

The objective of the study is to find out the effect of shark chondroitin on T lymphocyte subsets in cancer patients. Patients were divided into two groups. One group was treated with chemotherapy alone, and the other group was treated with chemotherapy plus shark chondroitin. Using immunofluorescence technique, T lymphocyte subsets in the peripheral blood were determined in two groups before and after chemotherapy. The results showed that CD4(+)/CD8(+) ratio increased in the patients received shark chondroitin. In the chemotherapy group, CD3(+) had no change, but CD4(+) decreased while CD8(+) increased significantly. The results suggest that shark chondroitin could enhance immune function in cancer patients, especially during chemotherapy.

This paper present a microcontroller system for target controlled infusion according to pharmacodynamic parameters of intravenous anesthetics. It can control the depth of anesthesia by adjusting the level of plasma concentrations. The system has the advantages of high precision, extended function and easy operation. It has been now used in the clinical anesthesia.
Algorithms ; Anesthesia, Intravenous ; instrumentation ; methods ; Anesthetics, Intravenous ; administration & dosage ; pharmacokinetics ; Computer Systems ; Humans ; Infusions, Intravenous ; Microcomputers ; Software Design

Objective To evaluate the role of PI3K∕Akt signaling pathway in dexmedetomidine-in-duced reduction of lung ischemia-reperfusion ( I∕R ) injury in rats undergoing cardiopulmonary bypass (CPB). Methods Twenty-four healthy adult male Sprague-Dawley rats, weighing 350-450 g, were di-vided into 3 groups (n＝8 each) using a random number table method: group I∕R, dexmedetomidine group ( group D) and dexmedetomidine plus wortmannin group (group D+W). Rats were anesthetized with pento-barbital sodium. Lung I∕R was induced by clamping the left hilum of lung for 60 min starting from 10 min of CPB, followed by 120-min reperfusion. Dexmedetomidine was injected via the tail vein in a dose of 3 μg∕kg at 10 min before clamping the left hilum of lung, followed by a continuous infusion of 1. 5 μg·kg-1·h-1 until the end of CPB in group D. Dexmedetomidine was injected via the tail vein in a dose of 3 μg∕kg at 10 min before clamping the left hilum of lung, followed by a continuous infusion of 1. 5 μg·kg-1·h-1until the end of CPB, and wortmannin was simultaneously injected via the tail vein in a dose of 15 μg∕kg, fol-lowed by a continuous infusion of 2. 0 μg·kg-1·min-1until the end of CPB in group D+W. Arterial blood samples were collected immediately before CPB ( T1), immediately after opening the left hilum of lung (T2) and at 1. 5 h after the end of CPB (T3), and oxygenation index (OI) and respiratory index (RI) were calculated. The rats were sacrificed at T3, and the left lung was removed for examination of the patho-logical changes which were scored and for determination of apoptosis rate ( by flow cytometry) and Akt, Bad, activated caspase-3, phosphorylated Akt ( p-Akt) and phosphorylated Bad ( p-Bad) in lung tissues ( by Western blot). Results Compared with the baseline at T1, OI was significantly decreased and RI was increased at T2and T3in the three groups (P＜0. 05). OI was significantly decreased and RI was increased at T3than at T2in the three groups ( P＜0. 05). Compared with group I∕R, OI was significantly increased and RI was decreased at T3, the pathological damage score and apoptosis rate were decreased, ratios of p-Akt∕Akt and p-Bad∕Bad were increased, and the expression of activated caspase-3 was down-regulated in group D, and OI was significantly decreased and RI was increased at T2in group D+W ( P＜0. 05). Com-pared with group D, OI was significantly decreased and RI was increased at T3, the pathological damage score and apoptosis rate were increased, ratios of p-Akt∕Akt and p-Bad∕Bad were decreased, and the ex-pression of activated caspase-3 was up-regulated in group D+W ( P＜0. 05). Conclusion Dexmedetomi-dine can reduce dexmedetomidine-induced reduction of lung I∕R injury through activating PI3K∕Akt signa-ling pathway and inhibiting cell apoptosis in rats undergoing CPB.

This study was aimed to investigate the factors influencing mobilization efficiency of peripheral hematopoietic stem cells with granulocyte colony stimulating factor (G-CSF) and their impact on healthy donors. 181 donors were mobilized subcutaneously with G-CSF at 5 - 10 µg/(kg·d), and 10 donors were mobilized with G-CSF at 3.3 - 4.9 µg/(kg·d), once 12 h, for 4 - 5 d. Peripheral blood mononuclear cell (MNC) and CD34(+) cell counts were analyzed by flow cytometry. Mobilization-related side effects were also monitored. The results showed that white blood cell counts increased by 6 times averaged after mobilization (P < 0.01). The platelet count obviously decreased (P < 0.01), while the hemoglobin level did not show significant difference. No significant differences were observed in MNC and CD34(+) cell counts between those subjects harvested on the 4th and 5th day after mobilization. Male donors were superior to female ones in cell harvest (P < 0.01). Donor body weight played positive role in cell yield, while impact of age on harvest was not remarkable. Neither MNC nor CD34(+) cell count showed a linear relationship with G-CSF dose. Only slight side effects were observed on the donors in this study. It is concluded that mobilization with G-CSF is sufficient in healthy donors without remarkable side effects.
Adolescent ; Adult ; Child ; Factor Analysis, Statistical ; Female ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; methods ; Tissue Donors ; Young Adult

The aim of this study was to investigate the feasibility of monitoring minimal residual disease (MRD) of leukemia with methylation specified-polymerase chain reaction (MS-PCR). The HL-60 cells with Id4 gene complete methylation and Hek937 cells with Id4 gene complete unmethylation were mixed in accordance with different ratios of cells and were divided into 3 groups: group A (10% HL-60 + 90% Hek937), group B (1% HL-60 + 99% Hek937) and group C (0.1% HL-60 + 99.9% Hek937). The MS-PCR technique was used to detect the methylation status of Id4 gene in different ratios of leukemia cells. The results indicated that the methylation specific amplification of Id4 gene with 155 bp was observed in HL-60 cells showing complete methylation of Id4 gene; while the unmethylation specific amplication of Id4 gene with 156 bp was found in Hek937 cells, showing complete unmethylation. The methylation specific amplification of Id4 gene with 155 bp and unmethylation specific amplification of Id4 gene with 156 bp were simultaneously detected in A, B and C groups, which showed the expression of Id4 gene methylation. In conclusion, the MS-PCR technique can detect the Id4 gene methylation in leukemia cell sample containing 0.1% of HL-60 cells, moreover the Id4 gene methylation in various leukemia cells shows no significant difference, thereby use of MS-PCR to detect the Id4 methylation level may be a potential approach for monitoring of MRD. Id4 gene promoter methylation is a candidate of biomarker for MRD detection in acute leukemias.
HL-60 Cells ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Leukemia ; diagnosis ; Methylation ; Neoplasm, Residual ; diagnosis ; Polymerase Chain Reaction ; methods