Objective To establish a method of inducing dendritic cells(DC)from rat bone marrow cells in vitro,and identify the phenotype and function characteristics.Methods The rat bone malToW cells were collected and cultured in vitro under the condition of recombinant rat GM-CSF(rrGM-CSF)and recombinant rat IL-4(rrIL-4).After 2 weeks,the morphological character of DCs was observed under light microscope and scanning electron microscope.Expression of MHC-Ⅱ,CD80 and CD86 were detected by flow cytometry.The ability to stimulate allogenic T cells of the cultured DCs was detected by mixed lymphocyte reaction.Results DCs showed typical morphology with elongated dendritic processes under inversion microscope and scanning electron microscope.DCs at day 6 revealed immature phenotype,including MHC-Ⅱ(29.03 ±4.39)%,CD80(21.98±7.08)%and CD86(25.94±6.80)%.DCs at day 12 showed higher expression of MHC-Ⅱ(74.05±5.97)%,CD80(79.85±6.53)%and CD86(81.00±7.47)%,and stimulatory capacity of allogenic T cells,compared with that in DCs at day 6.Conclusion Matured DCs could be generated from rat bone marrow cells and attendance with rrGM-CSF and rrIL-4,which present the feasibility for further research on its application to allograft immunorejection.

Objective To study the method of measuring the position accuracy and the step distance accuracy of afterloading system with192Ir source by using flushing-free film.Methods The position accuracy and the step distance accuracy of a China-made afterloading system with192Ir source was measured by using GAFCHROMIC (R) EBT3 flushing-free film.The film was scanned to proper image format,required by dose analysis software,by EPSON PREFACTION V700 PHOTO scanner.Then images are analyzed by using film dose analysis software in SNC Patient 5.2.Results With focus on the center of active section of source,the position accuracy of this afterloading system with192Ir source was-0.75 mm.Using film analysis could make the step point to tell apart if the step distance was 5 mm away by the method of film analysis,but couldnot make it to tell apart if the step distance was 2.5 mm away.The 2.5 mm step distance accuracy could be judged if the distance between the 1 st point and the 3rd point was 5 mm,then the 2.5 mm step distance could be deemed to no deviation.The 5 mm step distance of this afterloading system had no deviation in continuous 9 step points measured by flushing-free film.The indirect measuring results of the 2.5 mm step distance had no deviation as well.The position accuracy of this afterloading system measured with the flushing-free film accorded with the national standards.Conclusions The method of measuring the position accuracy and the step distance accuracy of the afterloading system with192Ir source by using flushing-free film is technically feasible.

The objective of this study was to investigate the efficacy and toxicity of standard-dose idarubicin in combination with continuous infusion of cytarabine as induction therapy in patients with acute myeloid leukemia (AML). A total of 38 AML patients were enrolled, including 30 new diagnosed patients, 8 relapsed and refractory patients. Cytogenetic analysis was performed in all patients, 15 patients had cytogenetic aberrations including 4 complex abnormalities. All patients were treated with standard-dose idarubicin [12 mg/(m(2).d), days 1 to 3] and continuous infusion of cytarabine [100 mg/(m(2).d), days 1 to 7]. The results showed that after one course of induction therapy, the overall response rate was 89.5% (34/38), and 32 out of 38 (84.2%) patients achieved complete remission (CR), including 27 of 30 (90.0%) new diagnosed AML patients, 5 (62.5%) refractory and relapsed AML patients, all 4 patients with complex cytogenetic aberrations achieved cytogenetic CR. Out of 6 relapsed patients 2 showed as extramedullary relapse, 4 showed as bone marrow relapse. The median survival duration was > 22 months and median disease-free survival time was > 16 months. Myelosuppression and infections due to neutropenia were the most frequent adverse effects, severe nonhematologic toxicity and the early death were not observed in the patients. It is concluded that standard-dose of idarubicin combined with continuous infusions of cytarabine as the induction therapy is highly effective and well tolerated approach in patients with AML, this regimen provides an opportune moment for hematopoietic stem cell transplantation.
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Cytarabine ; administration & dosage ; therapeutic use ; Female ; Humans ; Idarubicin ; administration & dosage ; therapeutic use ; Leukemia, Myeloid, Acute ; drug therapy ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

BACKGROUNDAs with many studies carried out in European countries, a quality assurance program has been established by the National Center for Clinical Laboratories in China (NCCL). The results showed that the external quality assessment significantly improves laboratory performance for quantitative evaluation of hepatitis C virus (HCV) RNA.

METHODSSerum panels were delivered twice annually to the clinical laboratories which performed HCV RNA detection in China. Each panel made up of 5 coded samples. All laboratories were requested to carry out the detection within the required time period and report on testing results which contained qualitative and/or quantitative test findings, reagents used and relevant information about apparatus. All the positive samples were calibrated against the first International Standard for HCV RNA in a collaborative study and the range of comparison target value (TG) designated as +/- 0.5 log.

RESULTSThe numbers of laboratories reporting on qualitative testing results for the first and second time external quality assessment were 168 and 167 in the year of 2003 and increased to 209 and 233 in 2007; the numbers of laboratories reporting on quantitative testing results were 134 and 147 in 2003 and rose to 340 and 339 in 2007. Deviation between the mean value for quantitative results at home in 2003 and the target value was above 0.5 log, which was comparatively high. By 2007, the target value was close to the national average except for the low concentrated specimens (10(3) IU/ml). The percentage of results within the range of GM +/- 0.5 log(10) varied from 8.2% to 93.5%. Some laboratories had some difficulties in the exact quantification of the lowest (3.00 log IU/ml) as well as of the highest viral levels (6.37 log IU/ml) values, very near to the limits of the dynamic range of the assays.

CONCLUSIONSThe comparison of these results with the previous study confirms that a regular participation in external quality assessment (EQA) assures the achievement of a high proficiency level in the diagnosis of HCV infection. During the 5-year external quality assessment, sensitivity and accuracy of detection in most of the clinical laboratories have been evidently improved and the quality of kits has also been substantially improved.

Hepacivirus ; genetics ; Humans ; Laboratories ; standards ; Polymerase Chain Reaction ; Quality Control ; RNA, Viral ; analysis ; Reagent Kits, Diagnostic

Stem cells use asymmetric and symmetric cell division to generate progeny. Symmetric cell division is defined as the generation of daughter cells that are destined to acquire the same fate. Stem cells divide asymmetrically to generate one daughter with a stem-cell fate and one daughter with different fate. Disruption of the machinery that regulates asymmetric division may be a reason for the generation of cancer. The asymmetric mechanism is maintained by cell polarity factors, cell fate determinants, and the spindle apparatus. The mutation or dysregulation of these factors may change stem cells from asymmetric to symmetric cell division, then leading to tumorigenesis. Therefore, further study is needed on the mechanisms of stem cell control between asymmetric and symmetric cell division, as well as the relationships among stem cells, cancer stem cells, and tumor cells. It may bring us a new approach for the resistance, recurrence, and metastasis of tumors.
Animals ; Cell Division ; physiology ; Cell Polarity ; Cell Transformation, Neoplastic ; Drosophila ; cytology ; Humans ; Neoplasms ; pathology ; Neoplastic Stem Cells ; pathology ; Neurons ; cytology ; Spindle Apparatus ; metabolism ; Tumor Suppressor Proteins ; metabolism

Objective To evaluate the demand for midwives in Yunnan province utilizing Birthrate Plus for planning and development of such workforce. Methods A convenient sample method was used to investigate 8435 maternal cases at 9 hospitals in Yunnan province in four months from 9 - 12 in 2017, and Birthrate Plus was used to calculate the demand for midwifery at each hospital. We also analyzed the two core elements of Birthrate Plus- maternal category allocation and midwife hours of each hospital. Results Maternal cases fall into five categories and maternal category allocation in hospitals is roughly the same;Maternal in higher category tended to need longer midwife hours; the average birthrate of 9 hospitals was (194. 22 ± 44. 84) case/ ( year·midwife). The number of midwives in two tertiary hospitals is obviously insufficient. Midwives at 7 secondary hospitals are more than predicted. Conclusions Midwives in Yunnan are generally faced with a large workload, especially at secondary hospitals, and midwives need to bear numerous non-midwifery workload beyond Birthrate Plus. Therefore, the Birthrate Plus can reflect the midwifery workload scientifically and reasonably in the current situation of midwifery work. But the predication for midwifery workforce requires a study of the ratio of midwifery work in the entire clinical work of the hospital.

This study was aimed to compare K562 cell proliferation, cell cycle and apoptosis before and after adhesive culture with MSCs of the same and different counts, so as to investigate the effect of human bone marrow mesenchymal stem cells (MSCs) on the growth of K562 cells. Culture system of bone marrow MSCs and co-culture system of K562 cells and BMSCs in vitro were established. And K562 cell proliferation curves were drawn by co-cultured of K562 cells with different counts of BMSCs. Cell cycle and apoptosis were determined by flow cytometry. The results showed that compared with the control, the proliferation of K562 cells cultured with the same amounts of MSCs was inhibited. After co-culture with the different amounts of MSCs, MSCs exhibited a dose-dependent anti-proliferation effect on K562 cells. The percentages of K562 cells in G(1) phase and G(2) phase were higher than those of the control. It is concluded that the normal bone marrow mesenchymal stem cells can inhibit the proliferation, the progress of cell cycle and the rate of apoptosis of K562 cells. As the number of mesenchymal stem cells increased, their anti-proliferation effect on the K562 cells were enhanced in a certain range.
Apoptosis ; Bone Marrow Cells ; cytology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Flow Cytometry ; Humans ; K562 Cells ; Mesenchymal Stromal Cells ; cytology

To evaluate the expression of cyclin dependent kinase inhibitor P27(Kip1) in leukemia and to investigate its clinical significance, the P27(Kip1) protein in bone marrow or peripheral blood samples from 82 cases of leukemia was measured by Western blot and enhanced chemoluminescence (ECL). The results showed that the expression of P27(Kip1) protein in ALL was higher than that in ANLL (P = 0.033) and also that in CML (P = 0.008). P27(Kip1) expression in CLL was higher than that in CML too (P = 0.017). In acute leukemia, the effective rate (CR and PR) of initial chemical therapy in the group of P27(Kip1) > 0.655 was higher than that in the group of P27(Kip1) < or = 0.655, P = 0.041. For ANLL and ALL patients, the survival time in the group of P27(Kip1) > 0.655 was longer than that in the group of P27(Kip1) < or = 0.655, P = 0.0065. There were similar statistical significance for ANLL and ALL patients, P = 0.0271 and P = 0.0266 respectively. There was a negative correlation between chromosomal abnormalities and P27(Kip1) expression in ALL patients (r = -0.775, P = 0.04). The expression of P27(Kip1) protein appeared nothing to do with sex, age, white blood cell number, blast cell number in peripheral blood, serum LDH or uric acid. In conclusion, the expression level of P27(Kip1) protein is in relation to the effect of initial chemical therapy and survival time, so that the lower P27(Kip1) expression may associated with poor prognosis in acute leukemia.
Adolescent ; Adult ; Aged ; Blotting, Western ; Cell Cycle Proteins ; analysis ; Child ; Child, Preschool ; Chromosome Aberrations ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; Leukemia, Lymphocytic, Chronic, B-Cell ; drug therapy ; genetics ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; metabolism ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; metabolism ; Survival Rate ; Tumor Suppressor Proteins ; analysis

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; therapy ; Peripheral Blood Stem Cell Transplantation ; Prognosis ; Retrospective Studies ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo analyze the expression of genes in the Slit/Robo signaling pathway, and the methylation status of their promoters in hepatocellular carcinoma (HCC) cell lines.

METHODSGenomic DNA and total RNA were isolated from 9 HCC cell lines of different metastatic ability (Hep3B, HepG2, PLC/PRF/5, SMMC-7721, BEL-7402, MHCC97-H, MHCC97-L, LM3, LM6) and a control cell line L-02. The expression profiles of Slit1, Slit2, Slit3, Robo1, and Robo3 were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The methylation status of the promoters was detected by methylation specific polymerase chain reaction (MSP).

RESULTSThe promoters of Slit1, Slit2 and Slit3 genes were almost methylated in all the HCC cell lines. The Slit1 and Slit3 RNAs were not detected in most of the cell lines. Furthermore, the mRNA Slit2 was decreased gradually as the metastatic potential of the cell lines increased. As the candidate ligand of the Slit2 gene, Robo1 was frequently methylated in HCC cell lines whereas its mRNA was detected in all of these cells except SMMC-7721, BEL-7402 and L-02. Robo3 was unmethylated in HCC cell lines while its mRNA was not detected in these HCC cell lines.

CONCLUSIONThe hypermethylation status of Slit/Robo signaling pathway related genes is a universal event in the HCC. The hypermethylation status of Slit1, Slit2, Slit3 genes associated with the loss of expression or reduced expression. Those data suggest that Slit/Robo pathway may play a significant role in the progress or metastasis of HCC.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; CpG Islands ; genetics ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Membrane Proteins ; genetics ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; Promoter Regions, Genetic ; Receptors, Immunologic ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction