Objective To observe the preventive effect of mannatide on infection and relapse of idiopathic thrombocytopenic purura(ITP).Methods One hundred and twenty children with ITP were randomly divided into mannatide treatment group and prednisone control group.Control group venous dexamethasone of 3 d;then treated by prednisone.Treatment group added mannatide tablets for 1 month.The rates of remission clinical blood,platelet,control time,complicated infection and relapse rates were observed.The levels of plasma immunoglobulin(Ig)G,IgA,IgM were determined before and after mannatide treatment.Results The rate of clinical blood,platelet,control time,infection time was not different in 2 groups.The rates of infection complicated and relapse were all significant lower than that in control group.The plasma IgG,IgA significantly increased than that in control group.The plasma IgM had no significant difference.Conclusion Vaccine therapy can be helpful in protecting and decreasing infection,diminishing relapse of children with ITP,and improve the level of IgG,IgA,and thus improve their immune function.

Objective To evaluate the theraputic effect of transcatheter arterial chemoembolization (TACE)for hepatocellular carcinoma with tumor thrombosis of portal vein.Methods One hundred and six patients of hepatocellular carcinoma with tumor thrombosis of portal vein under treament of TACE were observed before and after the procedure.Results After TACE tumor size reduced＞50% in 23 patients,＜50% in 25, no significant change in 44.The size of tumor enlarged in 12.The disappearance of portal vein tumor thrombosis accessed in 14,with reduction in 39,and no significant change in 51.Two patients died within one week.Conclusion TACE provides good therapeutic effect on hepatocellular carcinoma with tumor thrombosis of portal vein.

OBJECTIVETo explore the technique and clinical results of close reduction by manipulation and minimally invasive percutaneous osteosynthesis with intramedullary nail for the treatment of femur shaft fractures. methods: A retrospective study was conducted to analyze 96 patients with the femur shaft fractures who had been treated with close reduction by manipulation and minimally invasive percutaneous osteosynthesis with intramedullary nail. There were 67 males and 29 females. The average age of patients was 39 years old (ranging from 16 to 88). According to AO fracture classification for the femur shaft fractures,there were 29 cases of type A,46 type B,21 type C.

RESULTSAll the patients were followed up and the duration ranged from 12 to 24 months (averaged, 15 months). All the fractures showed union. The time required for the bony union ranged from 3 to 10 months (averaged,4 months). The clinical results were evaluated by Thorsen classification system. At the latest follow-up, 87 patients obtained excellent results, 7 good, 2 fair.

CONCLUSIONThis treatment method combines advantages of intramedullary nail with close manipulative reduction, so can get satisfactory clinical results for the treatment of femoral shaft fracture with minimal trauma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Nails ; Female ; Femoral Fractures ; diagnostic imaging ; surgery ; therapy ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; instrumentation ; Musculoskeletal Manipulations ; methods ; Retrospective Studies ; Tomography, X-Ray Computed ; Wound Closure Techniques ; Young Adult

OBJECTIVETo investigate the influence factors of adjacent segment degeneration (ASD) after instrumented lumbar fusion.

METHODSThirty-three patients who had undergone an instrumented lumbar fusion from March 1998 to May 2002 were reviewed. The incidence, age, position, radiographic characteristics and clinical manifestations of ASD were studied. Then the relations between "floating fusion" and ASD were compared, the range of fusion and ASD and investigated the incidences of different adjacent segments.

RESULTSThe mean follow-up period for the patients was 4 years and 7 months (24 - 82 months). Adjacent segment degeneration mainly occurred in patients older than 60. Ten patients (10%) were found to have radiographic characteristics of ASD. Nine of the ten patients had ASD at cranial segments. Using "floating fusion" or not did not show difference in the risk of ASD. There was a trend of more ASDs after long-segment fusion than short-segment fusion. As an adjacent segment, L(2)/L(3) had a high risk of ASD, while L(5)/S(1) had a much lower risk.

CONCLUSIONSThe cranial segment has a higher degeneration risk than the caudal segment. If L(2)/L(3) has degenerative appearance and has chance to be the adjacent segment, we'd better fuse it. If there is no evidence of obvious degeneration, L(5)/S(1) should not be fused. During instrumented lumbar fusion, long-segment fusion should be avoided if possible.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Diseases ; etiology ; Spinal Fusion ; adverse effects ; methods

OBJECTIVETo investigate the effect of PD98059 on the proliferation and cell cycle of rat hepatic stellate cells (HSCs) stimulated by acetaldehyde and explore its mechanism.

METHODSRat HSCs stimulated by acetaldehyde were incubated with different concentrations of PD98059. Cell proliferation was assessed by MTT colorimetric assay. Cell cycle was analysed by flow cytometry. The mRNA of cyclin D1 and CDK4 were examined by RT-PCR.

RESULTS20, 50, 100 micromol/L PD98059 could significantly inhibit the proliferation of HSCs stimulated by acetaldehyde in a does-dependent manner (0.109+/-0.020, 0.081+/-0.010 and 0.056+/-0.020 vs 0.146+/-0.030, F=31.385, P<0.05) and provoke G0/G1 phase arrest of HSCs stimulated by acetaldehyde in a does-dependent manner (61.9%+/-6.3%, 64.1%+/-3.3% and 70.9%+/-4.8% vs 55.2%+/-4.4%, F=16.402, P<0.05). 50, 100 micromol/L PD98059 could markedly inhibit cyclin D1 mRNA expression of HSC stimulated by acetaldehyde (0.56+/-0.04 and 0.46+/-0.03 vs 0.65+/-0.07, F=68.758, P<0.05) and CDK4 mRNA expression (0.39+/-0.07 and 0.33+/-0.05 vs 0.50+/-0.06, F=29.406, P<0.05).

CONCLUSIONThe Erk signal transduction pathway plays an important role in regulating the proliferation and cell cycle of rat hepatic stellate cells stimulated by acetaldehyde, which may be partly related to its regulative effect on the expression of cyclin D1 gene and CDK4 gene

Acetaldehyde ; pharmacology ; Animals ; Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinases ; metabolism ; Enzyme Inhibitors ; pharmacology ; Flavonoids ; pharmacology ; Hepatocytes ; drug effects ; Proto-Oncogene Proteins ; Rats

OBJECTIVETo construct the human/mouse chimeric antibody of a functional anti-death receptor 5 (DR5) antibody. Methods The viable region of light chain (VL) and viable region of heavy chain (VH) genes of anti-DR5 antibody were amplified and cloned into the light- and heavy-chain expression vectors respectively, then the recombinant plasmids were co-transfected into dihydrofolate reductase(-) Chinese hamster ovary cell (CHO-dhfr(-)) for expression. The positive clone was screened by the two selective genes (neo and dhfr). The humanization and specificity of chimeric antibody was identified by ELISA and Western blotting, and the tumoricidal activity of the expressed chimeric antibody was detected by tetrazolium salt phenazine methosulfate assay.

RESULTSThe expression vectors stably expressed chimeric antibody in CHO-dhfr(-). In the cell supernatant of the F4' clone, the human IgG heavy constant region and light constant region were identified. Moreover, the secreted chimeric antibody retained the binding capacity to the antigen (DR5) and decreased the cell viability of Jurkat and HCT116 cells to 73.15% and 77.30% in vitro respectively.

CONCLUSIONThe human/mouse anti-DR5 chimeric antibody has been successfully expressed in eukaryotic cells and shows tumoricidal activity, which establishes a foundation for the future research of humanized antibody medicine.

Animals ; Antibodies ; genetics ; immunology ; pharmacology ; Antineoplastic Agents ; immunology ; pharmacology ; CHO Cells ; Cell Survival ; drug effects ; Cricetinae ; Cricetulus ; Gene Expression ; Humans ; Mice ; Protein Engineering ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; immunology ; pharmacology

OBJECTIVETo identify the binding proteins to PDZ domain of ERBIN.

METHODSUsing PDZ domain of ERBIN as the bait, yeast two-hybrid technology was employed to screen the human lymphocyte leukemia cells MATCHMAKER cDNA library. The protein interaction was identified by immunoprecipitation.

RESULTSA PDZ-binding protein, TAX1, was identified.

CONCLUSIONTAX1 is a novel binding protein to PDZ domain of ERBIN.

Adaptor Proteins, Signal Transducing ; metabolism ; Cell Line, Tumor ; Humans ; Immunoprecipitation ; Intracellular Signaling Peptides and Proteins ; metabolism ; Neoplasm Proteins ; metabolism ; PDZ Domains ; Protein Binding ; Two-Hybrid System Techniques

OBJECTIVETo explore the molecular mechanism via which the chemotherapeutic drug hydroxyurea (HU) enhances K562 cell apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).

METHODSChronic myelogenous leukemia-derived K562 and SVT-35 cells were treated with recombinant soluble TRAIL (rsTRAIL) alone or combined with HU for a time course, and the cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-4-sulfophenyl-2H-tetrazolium-phenazine methosulphate assay. Western blot was performed to analyze the activation of apoptosis-related protein kinases and the expression of apoptosis inhibitor molecules.

RESULTSThe survival rates of SVT-35 and K562 cells treated with 1 μg/ml rsTRAIL for 24 hours were 32% and 93%, respectively. HU significantly increased the sensitivity of K562 cells to rsTRAIL cytotoxicity. Combination of rsTRAIL and HU resulted in the phosphorylation of rat sarcoma (RAS), mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase and in the significant reduction of apoptosis-inhibited molecule Fas associated death domain protein-like interleukin-1 beta-convening enzyme inhibitory protein and cellular inhibitor of apoptosis protein-1 in K562 cells.

CONCLUSIONSHU enhanced K562 cell sensitivity to rsTRAIL is mediated by Ras-MEK-ERK signaling pathway. Expression of antiapoptotic proteins cellular Fas associated death domain protein-like interleukin-1 beta-convening enzyme inhibitory protein and cellular inhibitor of apoptosis protein-1 is also down-regulated during this process. These results may through light on the therapeutic study of human chronic myelogenous leukemia.

Apoptosis ; drug effects ; physiology ; CASP8 and FADD-Like Apoptosis Regulating Protein ; metabolism ; Humans ; Hydroxyurea ; pharmacology ; Inhibitor of Apoptosis Proteins ; metabolism ; K562 Cells ; MAP Kinase Signaling System ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology

OBJECTIVETo identify the genes differentially expressed in leukemia cell apoptosis induced by recombinant soluble tumor necrosis factor-related apoptosis inducing ligand (rsTRAIL).

METHODSSuppression subtractive hybridization (SSH) and polymerase chain reaction (PCR) were used for the cloning and identification of the genes differentially expressed in the apoptotic Jurkat cells induced by TRAIL. Slot blot and Northern blot were used for the expression pattern analysis of the genes. Automatic DNA sequencing was used for DNA sequence analysis.

RESULTSSix cDNA fragments differentially expressed in the Jurkat leukemia cells treated with TRAIL were found, in which four were inhibited and two were activated during the Jurkat cell apoptosis treated with TRAIL. Among which the five genes of A14, X1, D1, A23 and C5 were found at the first time by DNA sequencing and GeneBank database searching. So that they were registered in GeneBank as AW731601, AW731602, AW731603, AW731604 and BE239235, respectively. It was found that the gene D1 was expressed higher in Jurkat leukemia cells and MCF-7 breast cancer cells than that in K562 leukemia, 825 gastric cancer and 7721 liver cancer cells.

CONCLUSIONSFive novel cDNA fragments were found, and among which D1 might be a tumor specific gene.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; genetics ; Apoptosis Regulatory Proteins ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Leukemia, T-Cell ; genetics ; pathology ; Ligands ; Membrane Glycoproteins ; pharmacology ; Polymerase Chain Reaction ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo clone and identify novel proteins binding to the death domain of the death receptor 4 (DR4).

METHODSThe yeast two-hybrid system was used for this study. Automatic sequencing was carried out for DNA sequencing. The sequence homology and the functional domains were analyzed by BLAST and the ScanProsite Tool softwares, respectively. Co-immunoprecipitate method was used to confirm human formyl peptide receptor-like 1 (FPRL1) binding specifically with DR4CD (the cytoplasmic domain of DR4) in HEK293T cells.

RESULTSTwo positive clones, named as pADB1 and pADB2, were obtained. BLAST searching showed that the homology of the insert sequence of pADB1 with the mRNA of FPRL1 was 97%. The insert of pADB2 shared no homology with any known peptides in GeneBank. Co-immunoprecipitate analysis further confirmed that FPRL1 could bind to DR4CD in vivo specifically.

CONCLUSIONSFPRL1 may associate with DR4CD in vivo specifically. The functional studies of FPRL1 in signaling pathway mediated by TNF-related apoptosis inducing ligand (TRAIL) are in active progress in our laboratory.

Amino Acid Sequence ; Apoptosis ; Apoptosis Regulatory Proteins ; Base Sequence ; Carrier Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Humans ; Membrane Glycoproteins ; metabolism ; Molecular Sequence Data ; Protein Structure, Tertiary ; Receptors, Formyl Peptide ; metabolism ; Receptors, Lipoxin ; metabolism ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; Receptors, Tumor Necrosis Factor ; genetics ; metabolism ; Signal Transduction ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Necrosis Factor-alpha ; metabolism