This paper expounds how the tractor for the fracture reduction works. The clinical results show that the traction apparatus is a labour-saving and time-saving orthopedic device with simple operation and few suffering to patients.
Arm Injuries ; diagnostic imaging ; surgery ; Equipment Design ; Fracture Fixation ; instrumentation ; methods ; Fractures, Bone ; diagnostic imaging ; surgery ; Humans ; Leg Injuries ; diagnostic imaging ; surgery ; Radiography ; Traction ; instrumentation ; methods

Objective: The current study was designed to investigate the relationship between bcl-2 expression and apoptosis of lung cancer cells in vitro. Methods:Lung cancer cell lines with different expression of bcl-2 were cocultured with tumor infiltrating lymphocyte(TIL) isolated from fresh tumor samples,and JAM test was performed to evaluate apoptosis of cancer cells. Immunohistochemistry and TUNEL assay were used to determine bcl-2 expression and apoptosis of tissue sections of lung cancers respectively. Results: As shown in JAM test, apoptosis increased with the elevation of TIL in coculturing assay in bcl-2 negative cancer cell lines, but in one of bcl-2 positive cell lines it remained stable. Conclusions: Lung cancer cells with bcl-2 expression may antagonize apoptosis, which may account for their immune evasion mechanism.

Cell Culture Techniques ; methods ; Fixatives ; Humans ; Immunohistochemistry ; methods ; Tissue Fixation ; methods

AIMTo explore a role of G6PD in replenishment of intracellular GSH during oxidative stress.

METHODSIn vitro Raji cell was cultured, intracellular GSH levels and G6PD, GR, GPX activities were determined at different time points after PMS treatment when G6PD activity was inhibited or not by DHEA.

RESULTSIntracellular GR, GPX, G6PD activities elevated significantly combined with GSH level decreased dramatically before 30 minutes, replenished gradually after 30 minutes and restore normal levels about 6 h after PMS treatment when G6PD was not inhibited. No change in GR and significant increase in GPX activity were shown following depleted GSH after PMS treatment when G6PD was inhibited by DHEA.

CONCLUSIONG6PD contributes to replenish intracellular GSH and is a critical factor regulating GSH levels during oxidative stress.

Cell Line, Tumor ; Glucosephosphate Dehydrogenase ; metabolism ; Glutathione ; metabolism ; Glutathione Peroxidase ; metabolism ; Humans ; Oxidation-Reduction ; Oxidative Stress ; Receptors, Peptide ; metabolism

OBJECTIVETo investigate the genes concerning multidrug resistance (MDR) of pancreatic ductal adenocarcinoma with microarray analysis.

METHODSGene expression profile of pancreatic cancer cell line SW1990 and resistance subline SW1990/5-FU, SW1990/ADM, SW1990/GEM were screened in two independent replicates using oligonucleotide microarray (Affymetrix HG U133 2.0 plus) which contained 38,500 human genes. And advanced bioinformatics analysis was conducted.

RESULTSTotally, 165 genes and expressed sequence tags (ESTs), which were seldom reported to be related with drug resistance before,were statistically difference and the fold change was up- or down-regulated at least 2 folds in all 3 resistant sub-lines when compared with SW1990. According gene ontology, the genes related to oxidoreductase activity, apoptosis, cell cycle, signal transduction and cell adhesion might be some epigenetic changes for MDR development. Hierarchical clustering analysis, showed several interesting clusters, namely, TNKS2, PRDX4 and CCDC4.

CONCLUSIONSMDR of pancreatic cancer is a complicated and multifactorial process. In the present study, a widespread differential gene expression pattern was constructed in PDAC multidrug resistant cells. Advanced study will provide new targets for MDR research and cast insights into research of the molecular mechanism of MDR.

Adenocarcinoma ; genetics ; pathology ; Carcinoma, Pancreatic Ductal ; genetics ; pathology ; Cell Line, Tumor ; Cluster Analysis ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Drug Screening Assays, Antitumor ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Neoplastic ; Humans ; Oligonucleotide Array Sequence Analysis ; Pancreatic Neoplasms ; genetics ; pathology

OBJECTIVETo screen the peptides that bind specifically to the hepatoma cells using phage display random peptides library.

METHODSThree rounds of panning were conducted in vitro targeting HepG2 cell lines. In nude mice bearing HepG2 tumor, one round of panning was conducted, and 30 phage clones were randomly selected for sequence analysis to identify the consensus sequence. Immunocytochemistry and immunohistochemistry were performed to determine the specificity of the phages to the hepatoma cells.

RESULTSAfter three rounds of panning in vitro and one round of panning in vivo, the phages binding to HepG2 cells were enriched. Sequence analysis of the randomly selected clones identified the peptide sequence VRKRSECLGAHD as the most frequent sequence. Immunocytochemistry and immunohistochemistry confirmed the specificity of the phage binding to the hepatoma cells.

CONCLUSIONSSpecific peptides against hepatoma cells can be obtained from a phage- display peptide library, which provides an experimental basis for developing therapeutic agents targeting hepatoma cells.

Animals ; Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Mice ; Neoplasm Transplantation ; Peptide Library ; Peptides ; isolation & purification ; metabolism ; Protein Binding

OBJECTIVETo study the effect of overexpression of Smad7 gene on cell proliferation in human bronchial epithelial cell lines.

METHODSHuman bronchial epithelial cell lines, BEP2D and BERP35T2 cells, were cotransfected with the mammalian expression vectors PCISmad7.neo and pMyc-SEAP, the latter was ac-myc cis-acting enhancer element fused with alkaline phosphatase (SEAP) reporter gene. Expression of c-myc, p15 and p21 mRNA was detected by RT-PCR before and after stable transfection of Smad7 into BEP2D and BERP35T2 cells in order to study the regulation of TGF-beta-mediated growth inhibition.

RESULTSAfter BEP2D and BERP35T2 cells transfected with Smad7, the transcriptional activity of c-myc was significantly increased. Smad7 overexpressing cells showed upregulation of c-myc expression and downregulation of p15 and p21 expression, which contributed to the loss of TGF-beta responses in these cells.

CONCLUSIONOverexpression of Smad7 may facilitate cell proliferation by antagonizing TGF-beta-mediated antiproliferative gene responses.

Bronchi ; cytology ; Cell Proliferation ; Cell Transformation, Neoplastic ; Cells, Cultured ; Cyclin-Dependent Kinase Inhibitor p15 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; genetics ; Epithelial Cells ; cytology ; Humans ; Proto-Oncogene Proteins c-myc ; biosynthesis ; genetics ; Signal Transduction ; Smad7 Protein ; biosynthesis ; genetics ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; genetics

BACKGROUNDChemotherapy is the most frequently adopted adjuvant therapy of pancreatic ductal adenocarcinoma (PDAC), but the development of drug resistance reduces its effectiveness. Clarification of the mechanism of multidrug resistance (MDR) development in PDAC is needed to improve the therapeutic effect of chemotherapy. This study was aimed to investigate the molecular mechanism of MDR of PDAC and to identify genes associated with MDR development.

METHODSThe gene expression profiles of cell line SW1990 and three drug-selected pancreatic chemoresistant sub-lines, SW1990/5-Fu, SW1990/ADM and SW1990/GEM, were obtained using an oligonucleotide microarray (Affymetrix HG U133 2.0 plus) that contained approximately 38,000 human genes. The microarray results were validated by real-time quantitative polymerase chain reaction and Western blot analysis.

RESULTSThere were 165 genes and expressed sequence tags, some of which have never been linked to drug resistance, that were up- or down-regulated at least 2-fold in all resistant sub-lines when compared with SW1990. According to Gene Ontology annotation, differentially expressed genes related to MDR in pancreatic cancer belong to many functional families and with diverse biological processes. Genes related to antioxidant activity, apoptosis, the cell cycle, signal transduction and intracellular adhesion may undergo epigenetic changes preceding MDR development. A hierarchical clustering was conducted and several interesting clusters were discovered that may be primarily related to cell cycle and developmental regulation. A prediction rule was built from the expression profiles of 117 genes after support vector machine (SVM) analysis, and the prediction result was examined by cytotoxic testing. As a result, a differential gene expression pattern was constructed in multidrug resistant pancreatic cancer cells.

CONCLUSIONSThe findings of this study prove that construction of a chemoresistance prediction rule, based on gene expression patterns, is practical. These data provide new insights into the molecular mechanism of pancreatic cancer MDR development and may be useful for the detection and treatment of MDR in pancreatic cancer patients.

Cell Cycle Proteins ; genetics ; Cell Line, Tumor ; Computational Biology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Gene Expression Profiling ; Glutathione Peroxidase ; genetics ; Glutathione Transferase ; genetics ; Humans ; Microtubule-Associated Proteins ; genetics ; Oligonucleotide Array Sequence Analysis ; methods ; Pancreatic Neoplasms ; drug therapy ; genetics ; Tankyrases ; genetics

OBJECTIVETo validate those obtained immunogenic membrane antigens candidate of human pancreatic cancer in the performed research.

METHODSIn the pre-studies, serum IgG purified from clinically collected sera of pancreatic cancer patients underwent immunoblot with human pancreatic cancer cell line SW1990 membrane protein, totally obtained 9 positive protein spots. Number 5 and 6 positive dots of immunoblot were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and peptide mass fingerprinting matching. The candidate membrane antigens were further validated in cell lines by RT-PCR and real-time PCR. RNA of human normal pancreatic tissue and pancreatic cancer tissue was extracted respectively, different gene expression level of prohibitin 2 was studied by real-time PCR.

RESULTSNumber 5 and 6 positive dots were identified as prohibitin 2 and prohibitin. RT-PCR and real-time PCR all showed that gene of prohibitin 2 and prohibitin were expressed in the human pancreatic cancer cell line SW1990, AsPc and P3 respectively, especially in P3 cell with highest expression (t = 7.442, P < 0.01). In addition, gene expression level of prohibitin 2 was significant higher in human pancreatic cancer than that of normal pancreatic tissue (t = 0.893, P < 0.01).

CONCLUSIONSProhibitin 2 and prohibitin are both differently expressed in the pancreatic cancer cell line SW1990, AsPc and P3. Prohibitin 2 is obvious highly expressed in human pancreatic cancer tissue. Prohibitin 2 and prohibitin might be the candidate immunogenic membrane antigens of human pancreatic cancer.

Antigens, Neoplasm ; genetics ; metabolism ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Membrane Proteins ; genetics ; metabolism ; Pancreatic Neoplasms ; genetics ; immunology ; metabolism ; Proteomics ; methods ; Repressor Proteins ; genetics ; metabolism

OBJECTIVETo study the expression and significance of GCS gene in human pancreatic cancer cell line SW1990 and its drug-resistant sublines.

METHODSSW1990 and its drug-resistant sublines, SW1990/FU, SW1990/ADM and SW1990/GEM were cultured in vitro. CCK-8 (Cell Counting kit-8) was used to detect the drug resistance of the sublines. Relative quantitation of GCS mRNA expression was evaluated by real-time PCR and Western blot was adopted to evaluate the expression of GCS protein.

RESULTSThe drug resistance indexes of SW1990/FU, SW1990/ADM and SW1990/GEM were 339.7, 11.9 and 56.6, respectively. The GCS mRNA and protein were expressed in all SW1990 and its drug-resistant sublines. There was a higher expression of GCS mRNA in all the sublines and a significant difference of GCS protein expression was detected in SW1990/ADM and SW1990/GEM compared with SW1990.

CONCLUSIONSGCS gene is expressed in SW1990 and its drug-resistance sublines. The high expression of GCS protein in SW1990/ADM and SW1990/GEM might be one reason of resistance to ADM and GEM in the sublines.

Cell Line, Tumor ; Drug Resistance, Neoplasm ; genetics ; Gene Expression ; Glucosyltransferases ; biosynthesis ; genetics ; Humans ; Immunoblotting ; Pancreatic Neoplasms ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction