OBJECTIVETo study the effects of minimal residual disease (MRD) level on day 33 of remission induction and IKZF1 genotype on the survival of children with B-lineage acute lymphoblastic leukemia (B-ALL).

METHODSA total of 152 children with newly-diagnosed B-ALL who had complete remission after the first cycle of the chemotherapy and had complete follow-up information were enrolled in this study. According to the MRD detection by flow cytometry on day 33 of remission induction, they were divided into three groups: standard-risk (SR) group (MRD <10; n=60), intermediate-risk (IR) group (10≤ MRD <10; n=55), and high-risk (HR) group (MRD ≥10; n=37). Nested RT-PCR was used to determine the IKZF1 genotype of all children before chemotherapy. The effects of MRD level on day 33 of remission induction and IKZF1 genotype on the recurrence-free survival (RFS) of children with B-ALL were analyzed.

RESULTSThere were 7 common IKZF1 subtypes in all the 152 children with B-ALL: IK1, IK2/3, IK4, IK6, IK8, IK9, and IK10. Of the 152 children, 130 had functional subtypes of IKZF1 and 22 had non-functional subtypes of IKZF1. During the follow-up period, relapse occurred in 26 (17%) children, and the recurrence rate was highest in the HR group (P<0.05). However, there was no significant difference in the recurrence rate between the SR group and the IR group (P>0.05). The cumulative recurrence rate of the children with non-functional subtypes of IKZF1 was significantly higher than that of those with functional types of IKZF1 (P<0.01). The predicted 5-year RFS rates in the SR, IR, and HR groups were (94.2±2.9)%, (86.7±3.8)%, and (56.2±4.5)% respectively (P<0.05). The 5-year RFS rate of the children with functional subtypes of IKZF1 was significantly higher than that of those with non-functional subtypes of IKZF1 (P<0.01). There was no significant difference in the predicted 5-year RFS rate between the children with functional subtypes of IKZF1 and those with non-functional subtypes of IKZF1 in the SR group (P>0.05). However, the predicted 5-year RFS rate of the children with functional subtypes of IKZF1 was significantly higher than that of those with non-functional subtypes of IKZF1 in the IR group and the HR group (P<0.05).

CONCLUSIONSB-ALL children with non-functional subtypes of IKZF1 have a high recurrence rate, and the recurrence rate will be even higher in B-ALL children with non-functional subtypes of IKZF1 and MRD ≥10 on day 33 of chemotherapy.

Antineoplastic Combined Chemotherapy Protocols ; Child ; Child, Preschool ; Female ; Genotype ; Humans ; Ikaros Transcription Factor ; genetics ; Male ; Neoplasm, Residual ; genetics ; mortality ; therapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; mortality ; therapy ; Prognosis ; Recurrence ; Remission Induction ; Survival

OBJECTIVETo select the best preparation method of vincristine transfersomes (VCR-T) and predict its possibility of being a new formulation of VCR.

METHODOrthogonal design was used to optimize the preparation methods on the basis of single factor pretests; and the permeation tests in vitro were performed in modified Franz diffusion cells.

RESULTThe optimum formula was: pH was equal to 7.3, the ratio of lecithin to sodium deoxycholate is 70/20, the weight of VCR is 10 mg, hydrating time is 30 minutes. The optimized solution was light yellow and transparent colloid solution. The VCR-T are spherical and smooth with average diameters of 94 nm and an encapsulation ratio of 90%. The test in vitro showed that VCR-T could permeat through mouse skin at zero rate with the cumulative penetrating quality amounting to 63.8%.

CONCLUSIONTransfersomes may become a promising carrier of VCR for clinic use.

Administration, Cutaneous ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacokinetics ; Deoxycholic Acid ; Drug Carriers ; Hydrogen-Ion Concentration ; Mice ; Particle Size ; Phosphatidylcholines ; Skin Absorption ; Technology, Pharmaceutical ; methods ; Vincristine ; administration & dosage ; pharmacokinetics

Background and Objectives: The effective use of MSCs for the treatment of some B cell-mediated immune diseases is quite limited. The main reason is that the immunomodulatory effects of mesenchymal stem cells (MSCs) on B cells are unclear, and their underlying mechanisms have not been fully explored. Methods: and Results: By co-culturing B cells with MSCs without (MSC/CTLsh) or with suppressor of cytokine signaling 1 (SOCS1) knockdown (MSC/SOCS1sh), we found that MSCs inhibited B cell proliferation, activation and terminal differentiation. Remarkably, the highest inhibition of B cell proliferation was observed in MSC/SOCS1sh co-culture. Besides, MSC/SOCS1sh reversed the inhibitory effect of MSCs in the last stage of B cell differentiation. However, MSC/SOCS1sh had no effect on inhibiting B cell activation by MSCs. We also showed that IgA+ B cell production was significantly higher in MSC/SOCS1sh than in MSC/CTLsh, although no difference was observed when both MSCs co-cultures were compared to isolated B cells. In addition, MSCs increased PGE2 production after TNF-α/IFN-γ stimulation, with the highest increase observed in MSC/SOCS1sh co-culture. Conclusions Our results highlighted the role of SOCS1 as an important new mediator in the regulation of B cell function by MSCs. Therefore, these data may help to develop new treatments for B cell-mediated immune diseases.

OBJECTIVETo investigate the change in dendritic cells (DCs) in children with chronic immune thrombocytopenia (cITP) and the effect of glucocorticoid on DCs in children with cITP.

METHODSFifteen children with cITP and 20 healthy controls were included in the study. Flow cytometry was used to measure the DC subsets count in the 15 children with cITP before and after glucocorticoid treatment as well as the corresponding values in the 20 healthy controls. The DCs derived from peripheral blood monocytes in children with cITP were cultured in vitro and collected, and their immunophenotypes were determined by flow cytometry.

RESULTSBefore glucocorticoid treatment, the children with cITP showed no notable change in the absolute count of myeloid DCs (mDCs) but showed decreased absolute count of plasmacytoid DCs (pDCs) and increased mDC/pDC ratio compared with the healthy controls (P<0.05). After glucocorticoid treatment, the children with cITP demonstrated increased absolute count of pDCs and decreased absolute count of mDCs and mDC/pDC ratio compared with before treatment (P<0.05). Before glucocorticoid treatment, the children with cITP had significantly higher positive rates of HLA-DR, CD80, CD83 and CD86 on peripheral blood DCs than the healthy controls (P<0.01). All the positive rates were significantly decreased after glucocorticoid treatment (P<0.01), so that there was no significant difference from the healthy controls (P>0.05).

CONCLUSIONSDisproportion and functional disturbance of DC subsets is associated with the pathogenesis of cITP in children. Glucocorticoid can strengthen the immunosuppression of DCs in children with cITP, which may contribute to the effectiveness of glucocorticoid as a treatment.

Adolescent ; Child ; Child, Preschool ; Chronic Disease ; Dendritic Cells ; drug effects ; immunology ; Female ; Glucocorticoids ; pharmacology ; Humans ; Immunophenotyping ; Male ; Thrombocytopenia ; drug therapy ; immunology

BACKGROUND AND OBJECTIVESince the proposal of the tumor stem cell hypothesis, considerable interest has been devoted to the isolation and purification of tumor stem cells. Tumor stem cell enrichment from primary tumor derived cell spheres has been demonstrated in specific, serum-free media. This goal of this study is to establish a method of cultivating floating tumor spheres from neuroblastoma cells and to confirm that neuroblastoma spheres are rich in tumor stem cells.

METHODSBone marrow aspirates were obtained from pediatric patients diagnosed with stage IV neuroblastoma. Primary tumor cells were isolated and cultivated in serum-free, stem cell-selective medium. Single sphere-forming cells were cultivated under serum-free conditions; their cloning efficiency and monoclonal tumor sphere formation rates were calculated. The expression of stem cell marker genes Oct-4 and Bmi-1 was detected by RT-PCR in sphere-forming cells and parental neurolastoma cells. Sphere-forming cells were injected into the armpit of nude mice with subsequent assessment for tumor growth. Sphere-forming cells were cultivated in differentiation medium containing 5 μmol/L 13-cis retinoic acid; changes in cell morphology were observed.

RESULTSNeuroblastoma cells formed non-adherent neurospheres under serum-free, stem cell-selective conditions after a period of 4 to 6 days. A single cell dissociated from a neurosphere could reform a monoclonal sphere; cloning efficiency and monoclonal sphere formation rates were 55.3% and 26.3%, respectively. RT-PCR results revealed heightened tumor sphere expression of Oct-4 and Bmi-1 as compared with parental tumor cells. Fourteen days after injection of 10(4) sphere-forming cells into nude mice, a neuroblastoma xenograft formed. Treatment of sphere-forming cells with 13-cis retinoic acid induced a gradual differentiation to neuronal cell morphology.

CONCLUSIONSNeuroblastoma derived tumor spheres enrich tumor stem cells and the cultivation of primary neuroblastoma cells in serum-free, stem cell-selective medium is an effective method to dissociate and purify tumor stem cells in vitro.

Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Child ; Culture Media, Serum-Free ; Humans ; Isotretinoin ; pharmacology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; metabolism ; pathology ; Neuroblastoma ; metabolism ; pathology ; Nuclear Proteins ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; metabolism ; Repressor Proteins ; metabolism ; Spheroids, Cellular ; pathology ; Xenograft Model Antitumor Assays

Objective To analyze the difference of teaching modes of course of digital signal processing for the undergraduates and postgraduates.Methods The teaching modes were compared from the aspects of educational objective,teaching content, teaching method,examination mode and etc.Results Differentiated teaching modes contributed to the satisfactory education of the undergraduates and postgraduates. Conclusion The differentiated teaching modes for the undergraduates and postgraduates provide references for the high-level education in universities and colleges of science and technology. [Chinese Medical Equipment Journal,2018,39(5):87-89,102]

AIM:To study the effect of Aurora protein kinase inhibitor VX-680 on homogeneous adhesion and migration ability in human hepatocellular carcinoma cell line HepG 2.METHODS:The HepG2 cell were divided into ex-perimental group and control group, respectively.VX-680 was used in experimental groups at 3 concentrations(3.125 μmol/L group,6.25 μmol/L group and 12.5 μmol/L group).DMSO was used in the control group.The effects of VX-680 at different concentrations on the adhesion ability of human hepatocellular carcinoma HepG 2 cells were observed by cell slow aggregation test and separation experiment.The effects of VX-680 at different concentrations on the migration ability of HepG2 cells was detected by wound healing assay.The expression of E-cadherin in HepG2 cells was detected by Western blot.RESULTS:The results of the slow aggregation test showed that compared with the control group,the number of cell clumps formed in experimental groups was significantly decreased(P<0.01).The results of separation experiment showed that the ratio of NTC/NTEgradually decreased with the increased concentration of VX-680.The results of wound healing as-say showed that as the concentration of VX-680 increased, the cell scratch healing ability gradually weakened compared with control group.The results of Western blot showed that the protein expression of E-cadherin in the HepG2 cells in-creased with the increased concentration of VX-680(P<0.05).CONCLUSION:VX-680 increases the homogeneous ad-hesion and inhibits the migration of HepG 2 cells.

OBJECTIVELcrV is an important component for the development of a subunit vaccine against plague. To reduce immunosuppressive activity of LcrV, a recombinant LcrV variant lacking amino acids 271 to 326 (rV270) was prepared by different methods in this study.

METHODSA new strategy that produced non-tagged or authentic rV270 protein was designed by insertion of rV270-thrombin-hexahistidine fusion gene into the vector pET24a, or by insertion of hexahistidine-enterokinase-rV270 or hexahistitine-factor Xa-rV270 fusion gene into the vector pET32a. After Co(2+) affinity chromatography, a purification strategy was developed by cleavage of His tag on column, following Sephacryl S-200HR column filtration chromatography.

RESULTSRemoval of His tag by thrombin, enterokinase and factor Xa displayed a yield of 99.5%, 32.4% and 15.3%, respectively. Following Sephacryl S-200HR column filtration chromatography, above 97% purity of rV270 protein was obtained. Purified rV270 that was adsorbed to 25% (v/v) Al(OH)₃ adjuvant in phosphate-buffered saline (PBS) induced very high titers of antibody to rV270 in BALB/c mice and protected them (100% survival) against subcutaneous challenge with 10⁶ CFU of Y. pestis virulent strain 141.

CONCLUSIONThe completely authentic rV270 protein can be prepared by using enterokinase or factor Xa, but they exhibited extremely low cleavage activity to the corresponding recognition site. Thrombin cleavage is an efficient strategy to prepare non-tagged rV270 protein and can be easily operated in a large scale due to its relatively low cost and high cleavage efficacy. The recombinant rV270 can be used as a key component to develop a subunit vaccine of plague.

Amino Acid Sequence ; Animals ; Antibodies, Bacterial ; blood ; Antigens, Bacterial ; genetics ; immunology ; Blotting, Western ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Female ; Genetic Vectors ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Plague ; immunology ; prevention & control ; Plague Vaccine ; genetics ; immunology ; Plasmids ; Pore Forming Cytotoxic Proteins ; genetics ; immunology ; Protein Engineering ; methods ; Recombinant Fusion Proteins ; genetics ; immunology ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Survival Analysis ; Vaccines, Subunit ; genetics ; immunology ; Yersinia pestis ; growth & development ; immunology

OBJECTIVETo understand the prevalence of HIV-1 drug resistance and its main factors in AIDS patients receiving to HAART in Zhecheng county, Henan province.

METHODSBy cross-sectional survey, 378 AIDS patients who had received the national free antiretroviral therapy from July 2003 to March 2009 in Zhecheng county of Henan were selected. All recruits were interviewed and blood samples were collected. HIV-1 viral loads were measured by real-time polymerase chain reaction, HIV genotypic resistance was determined by an in-house polymerase chain reaction to amplify the HIV-1 pol gene region.

RESULTSAmong 378 subjects receiving antiretroviral therapy, the prevalence of successful viral suppression was 42.9% (162/378), and HIV-1 drug resistance was detected in 47.1% (178/378) patients. The rate of resistance-associated mutations to non-nucleotide reverse transcriptase inhibitors (NNRTIs), nucleotide reverse transcriptase inhibitors (NRTIs) and protease inhibitors (PIs) was 46.6% (176/378), 37.8% (143/378) and 1.9% (7/378), respectively. Multivariate logistic regression analysis showed that the main risk factors associated with the prevalence of drug resistance included drug non-adherence (ratio of on-time drug intake in the past month < 90%:≥ 90% = 64.5% (40/62):43.7% (138/316), OR = 2.3, 95%CI: 1.3 - 4.1), and initiation of anti-retroviral therapy (ART) with didanosine (DDI) (ART with DDI:ART with lamivudine (3TC) = 53.3% (137/257):33.9% (41/121), OR = 2.3, 95%CI = 1.1 - 4.5).

CONCLUSIONThe prevalence of HIV-1 drug resistance among ART patients in Zhecheng county of Henan was high. Drug non-adherence and initiation of ART with DDI were the main factors of drug resistance.

Acquired Immunodeficiency Syndrome ; drug therapy ; virology ; Adolescent ; Adult ; Aged ; Anti-HIV Agents ; pharmacology ; therapeutic use ; Antiretroviral Therapy, Highly Active ; China ; Cross-Sectional Studies ; Drug Resistance, Viral ; Female ; HIV-1 ; drug effects ; Humans ; Male ; Middle Aged ; Young Adult

BACKGROUNDUnplanned extubation is associated with adverse outcomes in intensive care unit. The massive burn patient differs from other critically ill patients in many ways. However, little is known about the unplanned decannulation (UD) in Burn Intensive Care Unit. This paper describes the special features of the circumstances and outcome of UD of tracheotomy tube in massive burn patients.

METHODSA case series study was performed between January 1999 and December 2008 and UD of tracheotomy tube was analyzed retrospectively. A total of 21 patients with 29 UD events were identified. Demographic data, diagnosis, intervention, UD events and outcome of UD patients were collected. Differences in proportions were compared using the chi-square (χ(2)) or Fisher's exact test.

RESULTSPatients with UD were often burned with head and neck (67%) and combined with inhalation injury (62%). The majority of them (76%) were transferred patients, occurred early (55%) and were accidental UD (79%). UD events tended to happen in day shift (90%) and to be associated with the medical procedure that was performing by caregivers at besides (79%). Loose of the stabilizing rope, medical procedure and tracheotomy malposition were the main causes of UD. Early UD and reintubation failure were associated with patients' death.

CONCLUSIONSUD happened to massive burn patients can lead to patient death. Careful management of respiratory tract was essential for massive burn patients.

Adult ; Burns ; mortality ; surgery ; Device Removal ; adverse effects ; mortality ; Female ; Humans ; Intensive Care Units ; statistics & numerical data ; Intubation, Intratracheal ; Male ; Middle Aged ; Retrospective Studies ; Tracheotomy ; adverse effects