OBJECTIVECytokine mediated cell immunity is the main mode of anti-tumor immunity in organism, and the disequilibrium of cytokine network is the main cause of tumor cells escaping immunologic surveillance. High mobility group box 1 (HMGB1), a nuclear protein, has recently been identified as an important mediator of local and systemic inflammatory diseases when released into the extracellular milieu. In the present study, the investigators explored the clinical significance of alteration in the serum levels of HMGB1 in childhood acute lymphocytic leukemia (ALL) and the mechanism of HMGB1-induced tumor necrosis factor (TNF)-alpha secretion in leukemic cells.

METHODSThe serum levels of HMGB1 in healthy children and childhood ALL were assayed by Western blotting. K562 leukemic cells were stimulated with recombinant HMGB1 protein in vitro, and the secretion of TNF-alpha was determined by using ELISA. The effects of HMGB1 on activation of p38, c-Jun amino-terminal kinase (JNK), and extracellular-signal regulated protein kinase (ERK) and mitogen-activated protein kinase (MAPK) in K562 cells were assayed by using Western blotting. The effects of inhibitors specific for the MAPK on HMGB1-induced TNF-alpha secretion were assayed by using ELISA.

RESULTSThe serum levels of HMGB1 were significantly higher in ALL initial treatment group (n = 15, 43.78 +/- 4.62 microg/ml) than those in healthy control group (n = 15, 0.60 +/- 0.48 microg/ml, P < 0.01) and ALL complete remission group (n = 15, 0.89 +/- 0.62 microg/ml, P < 0.01). No significant difference was found between the healthy control group and ALL complete remission group in HMGB1 levels (P > 0.05). TNF-alpha started to become detectable at 2 h and was still increasing at 16 h after HMGB1 (1 microg/ml) treatment in K562 cell culture. TNF-alpha was also secreted from K562 cells in a dose-dependent manner after HMGB1 (1 ng/ml-1 microg/ml) exposure. HMGB1 induced the phosphorylation of p38, JNK and ERK in k562 cells. Inhibitors specific for the JNK (SP600125), MEK (PD98059), and p38 MAPK (SB203580), abrogated HMGB1-induced TNF-alpha secretion.

CONCLUSIONSThe measurement of serum HMGB1 is helpful to evaluate the prognosis of the childhood ALL. HMGB1 stimulates leukemic cells to secrete TNF-alpha through a MAPK-dependent mechanism.

Cell Line, Tumor ; Child ; Cytokines ; metabolism ; HMGB1 Protein ; metabolism ; Humans ; Imidazoles ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; Phosphorylation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; enzymology ; metabolism ; Protein Kinase Inhibitors ; pharmacology ; Pyridines ; pharmacology ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the effect of high mobility group boxl (HMGBI) gene silence on adriamycin (ADM)-induced apoptosis in K562/A02 drug resistance leukemia cells.

METHODSK562/ A02 cells were transient transfected with HMGB1- small interference RNA(siRNA) vector, and the levels of HMGB1 gene differential expression pre-and post-transfection were measured by RT-PCR and Western blotting. 50% inhibition concentration (IC50) of ADM on K562/A02 was determined by WST-8 assay. Cell apoptosis was assessed by flow cytometry. The release of Smac/DIABLO from the mitochondria to the cytoplasm was assayed by Western blotting. Activity of Caspase-3 was assayed with a Caspase Colorimetric Assay Kit.

RESULTS(1) The HMGB1 expression at mRNA and protein levels in HMGB1 siRNA transfected K562/A02 cells were decreased by 86% and 71% respectively compared with control. (2) Suppression of HMGB1 by siRNA in K562/A02 cells resulted in a reversal of the resistance to ADM, and decreased IC50 from (4.83 +/- 0.08) microg/ml to (1.33 +/- 0.10) microg/ml. 1 microg/ml and 5 microg/ml of ADM treatment increased cell apoptotic rate by 27% and 32% respectively. (3) HMGB1 suppression in K562/A02 cells significantly promoted ADM- induced Smac/DIABLO release from the mitochondria to the cytoplasm, and increased the activities of Caspase-3.

CONCLUSIONHMGB1 gene silence can enhance sensitivity of K562/A02 cells to ADM and reverse cell resistant to ADM.

Apoptosis ; drug effects ; genetics ; Doxorubicin ; pharmacology ; Gene Silencing ; HMGB1 Protein ; genetics ; Humans ; K562 Cells ; RNA, Small Interfering ; genetics

OBJECTIVETo evaluate the efficacy of salvaged allogeneic hematopoietic stem cell transplantation (allo-HSCT) for refractory/recurrent acute myeloid leukemia (AML).

METHODSA total of 45 patients with refractory/recurrent AML were enrolled from September 2006 to April 2010. The median blasts in bone marrow (BM) were 36% (20% to 92%) before conditioning. The donors were identical siblings (6) or unrelated ones (9) or haploidentical family members (30). Conditioning regiments were individualized according to patients' status, the regimen with high-dose cytarabine plus BuCy/CY was mostly used (20). The patients with impaired organ function received above regimen except using fludarabine instead of cyclophosphamide (16). FLAG followed by reduced-intensified BuCy was employed for the recipients with more than 40% blasts in BM (6) to reduce leukemia burden. TBI/CY or TBI/Fludarabine was used for the recipients with extramedullary infiltration of leukemia or multidrug resistant leukemia. G-CSF, MTX, NVT, Vm26, Acla or Thaltipa was added into conditioning regiments according to leukemia character.

RESULTSAll but 2 patients attained durable engraftment. The incidence of grade II to IV aGVHD and cGVHD were 34%, 59.1%, respectively. With median follow-up 30 (0.5 - 57) months, the relapse rate was 29.2%. Twenty-nine of 45 (60.2%) patients remained in complete remission since salvaged HSCT. Three-years disease-free survival and overall survival were 60.2% and 62.6%, respectively.

CONCLUSIONOur results indicated that the combination of salvaged HSCT with prophylactic immunotherapy might be a promising modality for treatment of refractory/recurrent AML, even with high leukemia burden.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia, Myeloid, Acute ; mortality ; therapy ; Middle Aged ; Recurrence ; Survival Rate ; Transplantation Conditioning ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo observe in vivo stem cell distribution and viability after transplantation by noninvasive imaging of 18F-fluorodeoxyglucose (18F-FDG) labeled autologous mononuclear bone marrow cells.

METHODSMyocardial infarction was established in 8 swine by ligating left anterior descending coronary artery after anesthesia. Bone marrow (20 ml) was drawn through ileum. After isolation, mononuclear bone marrow cells were labeled by radionuclide 18F-FDG and intramyocardially injected into infarction region. Whole body planar scan and myocardial tomography scan were performed immediately, 1 h, 2 h, and 3 h post stem cell injection. Viability and stability of radionuclide labeled stem cells were determined at 3 h post labeling in vitro.

RESULTSThe labeling efficiency was (67 +/- 14)%. Mean dose of radioactive in marrow cells was (32 +/- 7) MBq. Trypan blue staining showed in vitro viability was (95 +/- 3)% at 3 h post labeling. After intramyocardial injection, labeled mononuclear bone marrow cell retention rate in infarction region was (83 +/- 6)%, (49 +/- 8)%, (32 +/- 6)% and (24 +/- 5)% immediately, 1 h, 2 h, and 3 h post injection, respectively.

CONCLUSIONSDistribution and viability of stem cell after cardiac transplantation could be effective monitored by 18F-FDG labeled autologous mononuclear bone marrow cell technique in acute stage in this model.

Animals ; Bone Marrow Cells ; cytology ; diagnostic imaging ; Cell Survival ; Fluorodeoxyglucose F18 ; Graft Survival ; Heart Transplantation ; Myocardial Infarction ; diagnostic imaging ; surgery ; Radionuclide Imaging ; Stem Cell Transplantation ; methods ; Swine

In order to investigate the features of M-bcr/abl and m-bcr/abl fusion transcripts in patients with chronic myeloid leukemia (CML) after allogeneic stem cell transplantation (SCT), M-bcr/abl and m-bcr/abl fusion transcripts were sequentially detected by RT-PCR technique in 72 CML patients after SCT. The results showed that M-bcr/abl positive rate (79.2%, 42/53) within 6 months after SCT was remarkably higher than that in 6-12 months group (34.3%, 11/32) and >or= 12 months group (35.1%, 13/37) (P < 0.001), and the clinical relapse rates in corresponding periods were 1.9% (1/53), 0% (0/32) and 16.2% (6/37) respectively. M-bcr/abl and m-bcr/abl fusion transcripts occurred in 5 of 6 clinically relapsed patients. In period of more than 6 months after transplantation, none of 17 M-bcr/abl(+) samples from 14 patients in cytogenetic remission appeared positive reaction of m-bcr/abl. It is concluded that M-bcr/abl(+) fusion transcript still existed in most patients after SCT, and usually disappeared within 6 months. Existence of M-bcr/abl is not a clinical relapse marker in CML patients. Simultaneous detection of M-bcr/abl and m-bcr/abl fusion transcripts can be helpful for monitoring residual disease.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Fusion Proteins, bcr-abl ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; therapy ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Recurrence ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Homologous

OBJECTIVETo explore the effects of mitogen activated protein kinase (MAPK) and extracellular signal-regulated kinases (ERK) on kidney injury in female BALB/c mice exposed to cadmium.

METHODTwenty-one female BALB/c mice were randomly divided into 3 groups, i.e. control group, low Cd exposure group (2.5 µmol/kg) and high Cd exposure group (10 µmol/kg) were exposed to normal saline, 2.5, 10 µmol/kg Cd, respectively, 3 times a week for 14 weeks. The kidney slice were stained by HE, PAS and Masson staining to observe the morphological changes. The expression levels of pERK, ERK, pp38, p38, pJNK and JNK proteins in kidneys were tested by Western blot assay.

RESULTSThe ratios of pERK/ERK, pp38/p38, pJNK/JNK in high Cd group were higher than those in the control group (P < 0.05). The ratio of pERK/ERK in low Cd group was higher than control group (P < 0.05). The expression levels of bcl-2, bax proteins and the ratio of bcl-2 to bax in Cd exposure groups decreased significantly, as compared with the control group (P < 0.05). The impairment of renal glomeruli and tubules were observed in HE, PAS and Masson staining slices of kidneys in mice exposed to Cd.

CONCLUSIONCdCl2 may induced renal injury by affecting the expression levels of MAPK.

Animals ; Apoptosis ; drug effects ; Cadmium ; toxicity ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Kidney ; metabolism ; pathology ; MAP Kinase Signaling System ; Mice ; Mice, Inbred BALB C ; Mitogen-Activated Protein Kinases ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the unusual bcr/abl fusion gene structures of two Ph chromosome positive chronic myelogenous leukemia (CML) patients in chronic phase (CP).

METHODSBy using general M- and micro -bcr/abl specific primers respectively, bcr/abl fusion transcripts were detected by reverse transcription-polymerase chain reaction (RT-PCR). The RT-PCR products sequencing was performed, the DNA sequences were analyzed in Genebank and the bcr and abl sequences at the fusion site were identified. DNA was amplified by PCR using a set of primers designed according to the sequencing result of RT-PCR products.

RESULTSTwo patients showed typical manifestations of CML-CP. Their RT-PCR products were different from usual M- or micro -type; one was longer than M-bcr/abl but shorter than micro -bcr/abl, the other one was shorter than M-bcr/abl. The RT-PCR products sequencing showed that both products contained bcr and abl gene sequences. The first patient's bcr gene was broken within exon 18, and fused to abl gene exon 2(a2), and a 40 bp of partial abl intron 1b fragment was inserted between them, resulting in a novel in-frame bcr/abl fusion transcript-e18-int-a2 which has not been reported in the literature so far. In the second patient, deletion of abl exon2(a2) led to exon 13(b2) of bcr gene fusing with abl exon 3(a3).

CONCLUSIONUncommon bcr/abl fusion gene may occur in typical Ph(+) CML patient.

Adult ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Molecular Sequence Data ; Philadelphia Chromosome ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo investigate the relationship between three types of bcr/abl fusion transcripts and clinical manifestation in chronic myeloid leukemia (CML).

METHODM-, m- and micro -bcr/abl fusion transcripts were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) technique in 537 fresh bone marrow samples of patients suspected CML clinically.

RESULTSOf 573 patients, 479 expressed M-bcr/abl transcripts, among whom 370 were in chronic phase (CP), and 109 in accelerated (AP)/blastic phase (BP). The percentages of patients with b2a2 transcripts in CP and AP/BP were 32.4% (120/370) and 36.7% (40/109) (P > 0.05). The b2a2 transcript patients in blastic crisis were 52.6% (10/19) for lymphoblastic and 33.3% (30/90) for myeloblastic (P > 0.05). The platelet count of untreated patients with b3a2 isoform [(485.9 +/- 333.8) x 10(9)/L, n = 125] was distinctly higher than those with b2a2 isoform [(380.5 +/- 321.9) x 10(9)/L, n = 62] (P < 0.05). 66.0% (31/47) and 64.4% (29/45) of the patients in CP and AP/BP respectively co-expressed M- and m-bcr/abl transcripts (P > 0.05). One patient expressed only m-bcr/abl transcript was of typical acute myeloblastic leukemia (AML). Both two micro -bcr/abl(+) patients were of typical CML.

CONCLUSIONSAlmost all typical CML patients express M-bcr/abl transcripts, most of them coexpress M-bcr/abl and m-bcr/abl transcripts, a few possesses only micro -bcr/abl fusion gene. m-bcr/abl(+) are usually associated with AML or CML in myeloblastic crisis besides acute lymphoblastic leukemia (ALL). Patients with b3a2 isoform are prone to higher platelet count before treatment.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Fusion Proteins, bcr-abl ; genetics ; Genotype ; Humans ; Infant ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics

Objective?To analyze the clinical efficacy and related X-ray findings of patients underwent arthroscopic treatment of femoral acetabular impingement (FAI) syndrome with different anatomical features.?Methods?Twenty-four patients with FAI underwent arthroscopic surgery from September 2015 to December 2016 were selected to analyze the clinical features, postoperative pain, knee joint function, activity and complications.?Results?Compared with those before treatment, the visual analogue scale (VAS) scores of patients at 7 d, 1 month, 3 months and 6 months after treatment were significantly lower than those before treatment, while the Harris scores were significantly increased, at the same time patients’ knee activity was significantly increased, The difference was statistically significant (P < 0.05). The α angle of the hip joint of the cam-type patient was significantly higher than that of the jaw-type, while the eccentricity, acetabular depth, acetabular coverage and centerline (CE) angle were significantly lower than the jaw-type, and the difference was statistically significant (P < 0.05); The α angle of the cam-type patient was significantly higher than that of the healthy person’s hip joint, and the eccentricity was significantly lower than that of the healthy person’s hip joint. The difference was statistically significant (P < 0.05); the acetabular depth and hip of the clamp-type patient Radon coverage and CE angle were higher than those of hip joints in healthy people, and the difference was statistically significant (P < 0.05). There was no significant difference in acetabular anterior tilt between the three groups (P > 0.05). The incidence of complications in 24 patients underwent arthroscopy was 20.83%.?Conclusion?Hip arthroscopic treatment of hip impingement syndrome can shorten the patient’s pain relief, improve knee function and activity, its effect is good, and different hip anatomical X-ray film was significantly different.

Objective To improve the CR image quality and to promote the digital image standard constitution by analyzing the common problems and malpractiees in application of computed radiography. Methods Phenomenon and reasons of 107 CR junk-films from nine three-"A"-hospitals were analyzed, discussed,recorded,and statistised by 20 radiologists,radiographers and engineers.Results Among 107 junk fihns,there are 36 cases(33.64%)of incorrect operations,29 cases(27.10%)of artifacts in reading and transferring the data of IP,15 cases(14.02%)of artifacts in IP system,and 13 cases (12.15%)of selection of inappropriate radiographic parameters,and 9 eases(8.41%)of printer-failures, and 5 eases(4.67%)of inappropriate post-processing techniques.By analyzing the reasons of 107 junk films we found that 60.74% were due to less responsibilities and incorrect operations,and 35.51% were due to new problems in CR techniques,and other were due to inappropriate post-processing techniques. Conclusion Responsibilities,operation regulations,digital image quality standards,studying of new techniques and appropriate use of the post-processing techniques are the key points for improving the CR image quality and the diagnosis level.