Predicting liver metastasis of gastrointestinal tract cancer by diffusion-weighted imaging of apparent diffusion coefficient values.

Aim: To determine if efficacy of chemotherapy on liver metastasis of gastrointestinal tract cancer can be predicted by apparent diffusion coefficient (ADC) values of diffusion-weighted imaging (DWI).

Methods: In total, 86 patients with liver metastasis of gastrointestinal tract cancer (156 metastatic lesions) diagnosed in our hospital were included in this study. The maximum diameters of these tumors were compared with each other before treatment, 2 wk after treatment, and 12 wk after treatment.

[Molecular mechanism of hydroxyurea enhances K562 cell apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand].

Objective: To 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).

Methods: Chronic 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.

[Expression and tumoricidal activity of a human-mouse chimeric anti-DR5 antibody through a Furin/2A peptide].

Objective: To express a full-length human-mouse chimeric anti-DR5 antibody from a single open reading frame with tumoricidal activity to various cancer cells.

Methods: The heavy and light chains of chimeric antibody were joined by the Furin and 2A (F/2A) self-cleavage peptide and cloned into a lentiviral vector of pWPXL. Then the HEK293 cells were infected with the constructed expression vector pWPXL-HF2AL.

[Effect of miR-146a on IL-18 expression in mouse macrophage].

Aim: To investigate the effect of miR-146a on the Th1/Th2 cytokine expression in mouse RAW264.7 cell line and primary peritoneal macrophage.

Methods: miR-146a mimics, mimics negative control (NC mimics), inhibitor miR-146a and inhibitor negative control (NC inhibitor) were transfected into RAW264.

[Construction and stable expression of anti-human tumor necrosis factor-related apoptosis-inducing ligand receptor 2 chimeric antibody in CHO cells].

Aim: To establish an human-mouse chimeric antibody against tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor 2 (death receptor 5, DR5) in an eukaryotic cell line and analyse its tumoricidal activity.

Methods: The cDNAs encoding for the variable regions of heavy chain (V(H);) and light chain (V(L);) of AD5-10 were amplified by PCR and inserted into the human IgG heavy and light chain containing expression vector RpCI-neo, respectively. The recombinant plasmids were co-transfected into HEK293 and/or CHO cells.

[Killing effects of controllable expression of tumor necrosis factor-related apoptosis-inducing ligand from mesenchymal stem cells on hepatocellular carcinoma cells].

Objective: To study the controllable expression of soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in mesenchymal stem cells and evaluate its potential tumoricidal effects in cancer therapy.

Methods: The controllable TRAIL expression vector of Ad-Tet-TRE-TRAIL was established in an adenovirus vector for transfection into murine mesenchymal stem cells. The controllable expression and secretion of TRAIL were detected by Western blot and enzyme-linked immunosorbent assay.

Structural and functional analysis of the N-terminal region of death receptor 5.

Objective: To investigate the structure and function of the N-terminal region (NTR) of death receptor 5 (DR5).

Methods: A series of deletions of the DR5 extracellular domain (DR5-ECD) proteins were expressed in E.coli.

[Construction and expression of anti-tumor necrosis factor related apoptosis-inducing ligand receptor death receptor 5 chimeric antibody in eukaryotic cells].

Objective: To 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).

[Effects of human telomerase reverse transcriptase promoter and survivin promoter in targeted tumor gene therapy].

Objective: To investigate the effects of human telomerase reverse transcriptase (hTERT) promoter and survivin promoter in tumor-specific gene therapy.

Methods: hTERT promoter and survivin promoter were obtained by PCR using Jurkat genomic DNA. Recombinant adeno-associated virus (AAV) vectors containing exogenous TRAIL gene and hTERT promoter or survivin promoter were constructed and designated as rAAV-hTERT-TRAIL (h/TRAIL) or rAAV-survivin-TRAIL (s/TRAIL).

[Identification of binding proteins to the PDZ domain of ERBIN].

Objective: To identify the binding proteins to PDZ domain of ERBIN.

Methods: Using 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.

[Mechanisms of sensitization by 8-chloro-adenosine of human tumor cells to TRAIL-induced apoptosis].

Objective: To study the effect of 8-chloro-adenosine (8-Cl-Ado) on the sensitivity of human hepatoma and breast cancer cell lines to TRAIL-induced apoptosis in vitro and its mechanisms.

Methods: Recombinant soluble TRAIL (rsTRAIL) or 8-Cl-Ado was used to treat hepatoma cell line BEL-7402 and breast cancer cell line MCF-7 in vitro. MTT assay was used to evaluate cell viability.

[Establishment of high-throughput drug screening model targeting retinoic acid receptor in cells].

Aim: To screen new drug for the treatment of acute promyelocytic leukemia, psoriasis and acne, high-throughput drug screening cell models marked by green fluorescent protein (GFP) have been established.

Methods: Eight repeats of retinoic acid response element (RARE) were synthesized and cloned into a GFP expression vector. This construct was stably transfected into cells in vitro.

Evaluation of sequest result filter-Xcorr and Unified Score.

Objective: To estimate the effect of two simple filters, two or more positive peptide filter and Unified Score filter on the true positive rate of protein and peptide.

Methods: Twenty-two LC-MS/MS datasets were from 18 known protein mixture. Two or more positive peptide filter and Unified Score filter were applied to the 22 datasets.

[Expression of TRAIL(114-281) mediated by adeno-associated virus and its tumoricidal activity].

Objective: To investigate the expression of the soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) mediated by adeno-associated virus (AAV) and its tumoricidal activity in vitro and vivo.

Methods: The recombinant AAV expression vector encoding the extracellular domain (114-281aa peptide, TRAIL(114-281)) of TRAIL was constructed and transfected into human embryotic kidney cells HEK293 for virus package. The human tumor cell lines of T lymphocyte leukemia Jurkat, liver cancer HepG2 and SMMC-7721, and cervical cancer HeLa were transduced by using the recombinant virus particles respectively.

[Chemotherapeutic drugs enhanced rsTRAIL tumoricidal activity].

Objective: To explore the role and mechanisms of chemotherapeutic drugs in TRAIL induced cell death.

Methods: Tumoricidal activities of the chemotherapeutic drugs and/or rsTRAIL in 13 strains of tumor cell lines were evaluated by MTS-PMS assay and flow cytometry. DR5 expression in the cells was observed by Western blot.

[Synergetic role of CD3epsilon and 5-fluorouracil in T Lymphocyte apoptosis and P53 expression].

Objective: To investigate the relationship between apoptosis induced by CD3epsilon and 5-fluorouracil (5-FU), and study the P53 expression in the apoptosis process provide a novel insight and useful information of the apoptosis signaling pathway induced by CD3epsilon and/or 5-FU, and an important implication for the treatment of T-lymphocyte leukemia.

Methods: The viabilities of Jurkat T lymphocytes (JK), TJK [JK with over-expression of the CD8epsilon chimeria molecule] and T3JK [JK with over-expression of the CD8epsilon (Y170F/Y181F) mutation molecule] cells were cultured and treated with pre-coated anti-CD8 mAb (200 micro g/ml) and/or 5-FU (2.5 micro g/ml) were detected with MTS assay and the apoptosis percentages were calculated.

[The primary study on a novel protein binding to the death domain of the death receptor 4].

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

Methods: The yeast two-hybrid system was used for this study. Automatic sequencing was carried out for DNA sequencing.

[Differential expression of the genes in leukemia cell apoptosis induced by TRAIL].

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

Methods: Suppression 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.