OBJECTIVETo study the relationship between activation of pro-MMP-2 and expression of matrix metalloproteinases (MMP)-2, MT1-MMP and tissue inhibitor of metalloproteinases (TIMP)-2 mRNA in thymoma and thymic carcinoma; and to study the molecular mechanism of invasion and metastasis of thymic epithelial tumors.

METHODSFresh tissue specimens of thymoma, thymic carcinoma and normal thymus were included. The mRNA expression of MMP-2, MT1-MMP and TIMP-2 were analyzed by real-time reverse transcription polymerase chain reaction. The pro-MMP-2 activation ratio and its localization were determined by gelatin zymography and film in-situ gelatin-Zymography, respectively. Correlation of mRNA expression of MMP-2, MT1-MMP and TIMP-2 was investigated in tumors with different histological subtypes and clinical stages.

RESULTSThere were no significant differences in the expressions of MMP-2, MT1-MMP and TIMP-2 mRNA between I and II stage or III and IV stage thymomas (P > 0.05). However, significant differences of the expressions were observed between three tumor groups: I-II stage, III-IV stage and thymic carcinomas (P < 0.005), and between three histological subtypes: AB-B1 (lymphocyte-rich and mixed types), B2-B3 (cortical and predominantly polygonal cells types) and thymic carcinomas (P < 0.05). Expression levels of MT1-MMP and TIMP-2 mRNA were correlated with pro-MMP-2 activation ratio (Spearman rank correlation: r = 0.7235, r = 0.7647, P < 0.005). The expression of MMP-9 did not show significant differences between thymomas and thymic carcinomas.

CONCLUSIONSMMP-2, MT1-MMP and TIMP-2 mRNA expression levels are correlated with the histologic subtypes and clinical stages of thymoma. The mRNA expressions of MT1-MMP and TIMP-2 are correlated with the activation ratio of pro-MMP-2. It is speculated that upregulation of MT1-MMP gene expression may induce an activation of pro-MMP-2 through TIMP-2.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; enzymology ; metabolism ; pathology ; Enzyme Activation ; Female ; Humans ; Male ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; Matrix Metalloproteinases ; biosynthesis ; genetics ; Matrix Metalloproteinases, Membrane-Associated ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; biosynthesis ; genetics ; Thymoma ; classification ; enzymology ; metabolism ; pathology ; Thymus Gland ; enzymology ; metabolism ; Thymus Neoplasms ; classification ; enzymology ; metabolism ; pathology ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics

OBJECTIVETo investigate the protein expression of Axin and beta-catenin, the exon 3 mutation status of beta-catenin and their clinicopathological correlations in non-small cell lung cancer (NSCLC).

METHODSA total of 100 NSCLC samples and their corresponding normal lung tissues were obtained from the patients undergoing surgery in the First Affiliated Hospital of China Medical University between 2001 and 2003. Protein expressions of Axin and beta-catenin were detected by immunohistochemistry. DNA sequence alterations of exon 3 of beta-catenin were investigated by polymerase chain reaction (PCR) and direct sequencing.

RESULTSA reduced membranous expression rate of beta-catenin was observed in 80.0% of the cases (80/100) along with a nuclear expression rate of 26.0% (26/100). There was a significant difference in beta-catenin expression between well and poorly differentiated NSCLCs. Well to moderately differentiated NSCLCs showed a reduced expression rate of 70.0% (35/50), in contrast to 90.0% (45/ 50) in poorly differentiated tumors (P = 0.012). Reduced beta-catenin expression rate was 87.3% (48/55) in cases with lymph node metastasis, in contrast to 71.1% (32/45) in cases without lymph node metastasis (P = 0.044). The positive expression rate of Axin was 48.0% (48/100). Well to moderately differentiated NSCLCs demonstrated a 60.0% positive expression rate of Axin (30/50), much higher than poorly differentiated tumors [36.0% (18/50), P = 0.016]. The positive expression rate of Axin in beta-catenin nuclear expressed NSCLCs was 15.4% (4/26), much lower than cases without beta-catenin nuclear expression [59.5% (44/74), P < 0.001]. Axin nuclear expression was found in two cases in this study, suggesting that it may function as a nuclear-cytoplasmic shuttling protein. PCR and direct sequencing failed to reveal any exon 3 mutation of beta-catenin gene.

CONCLUSIONSThe reduced membranous expression of beta-catenin is associated with poorly differentiated and lymph node positive NSCLCs. The expression of Axin is inversely correlated with the degree of tumor differentiation and nuclear expression of beta-catenin. The exon 3 mutations do not contribute to the abnormal protein expression of beta-catenin in NSCLCs.

Adult ; Aged ; Axin Protein ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Differentiation ; Exons ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Mutation ; Neoplasm Staging ; Repressor Proteins ; metabolism ; beta Catenin ; genetics ; metabolism

OBJECTIVETo investigate the expression of p120(ctn) in non-small cell lung cancer (NSCLC) and its correlation with the clinical and pathologic parameters.

METHODSImmunohistochemistry (S-P method) was used to detect the expression of p120(ctn) in 143 NSCLC cases. The variation of protein expression was further analyzed in 36 cases by Western blot. The correlation with clinical and pathologic parameters was studied.

RESULTSImmunohistochemically, normal bronchial cells showed membranous expression for p120(ctn), while NSCLC was characterized by cytoplasmic or diminished membranous staining. The rate of abnormal p120(ctn) expression was 79.7% (114/143). There was a significant correlation between abnormal expression of p120(ctn) and tumor differentiation, clinical stage, lymph node metastasis and poor prognosis (< 0.05), but not histologic typing. Western blot showed that the total amount of p120(ctn) in normal bronchial cells was significantly higher than that in NSCLC. The p120(ctn) isoform 1 (120,000) and isoform 3 (100,000) were expressed in normal lung tissue, while there was a reduced expression or absence of isoform 1 in NSCLC.

CONCLUSIONThe expression of p120(ctn) is abnormal in NSCLC; p120(ctn) may serve as a useful prognostic marker for NSCLC.

Aged ; Biomarkers, Tumor ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Catenins ; Cell Adhesion Molecules ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung ; metabolism ; pathology ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Phosphoproteins ; metabolism ; Prognosis ; Proportional Hazards Models

Based on the stakeholder theory,externalities theory and marginal utility theory,this paper analyzes the behavioral needs of stakeholders in the process of market access of innovative drugs.It also draws out the core of the government and the pharmaceutical enterprises in the policy of access to innovative drug market and supply to the community,the patients,and the medical institutions enter the mechanism of the interaction of the various stakeholders in the innovative drug market for the demand community and construct the above-mentioned stakeholder perspective Innovative Drug Market Access Policy Environment Model.Based on the status quo of China's innovative drug market access,the present study puts forward to encourage innovative drug market access to the interests of the main body,to optimize the existing innovative drug market access policy environment to make reference recommendations.

OBJECTIVETo investigate the influence of exposure to different concentrations of ethanol on neural progenitor cells and the differentiation of neurons and glial cells in zebrafish embryos.

METHODSZebrafish embryos were exposed to 1%, 2%, and 2.5% (V/V) ethanol at 5 hpf by adding ethanol to the egg water. In situ hybridization and real-time PCR were used to detect the changes in the mRNA expression profiles of the markers of different cells to examine the effects of alcohol on neural development.

RESULTSThe number of neural precursor cells, neurons and mature glial cells was significantly reduced in the zebrafish embryos following ethanol exposure, and this reduction became more prominent as the ethanol concentration increased. The expression of the early glial marker slc1a3a was down-regulated in the spinal cord but increased in the brain after exposure to increased ethanol concentrations. The expression of the mature glial markers was significantly lowered in response to exposure to increasing ethanol concentrations.

CONCLUSIONSEthanol can reduce neural precursor cells and inhibits neuronal and glial differentiation in zebrafish embryos.

Animals ; Brain ; Cell Differentiation ; drug effects ; Embryo, Nonmammalian ; drug effects ; Ethanol ; adverse effects ; Neural Stem Cells ; drug effects ; Neurogenesis ; drug effects ; Neuroglia ; drug effects ; Neurons ; drug effects ; Spinal Cord ; Zebrafish ; embryology

Adolescent ; Adult ; Aged ; Child ; Cranial Nerve Neoplasms ; diagnosis ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neurilemmoma ; diagnosis ; surgery ; Radiosurgery ; Treatment Outcome ; Trigeminal Nerve Diseases ; diagnosis ; surgery

OBJECTIVETo retrospectively evaluate the effects of Gamma knife in the treatment of cerebral hemangioblastomas.

METHODSFrom 1993 to 1996, seventeen patients with 29 hemangioblastomas were treated with Gamma knife. The patients mean age was 35 years (range: 16 - 61 years). The mean tumor diameter was 16 mm (range: 6 - 55 mm). Thirteen patients had recurrent or residual hemangioblastomas. Four with primary hemangioblastomas were diagnosed using CT, MRI and DSA. The maximum dose to the tumors was 21.0 - 50.0 Gy, with mean dose of 33.7 Gy. The radiation dose to the periphery of tumors was 12.0 - 24.0 Gy, with mean dose of 17.6 Gy.

RESULTSAll the patients had been followed up for 18 to 62 months, with mean 46 months. Five patients experienced clinical improvement and reduction in tumor volume, and 5 remained stable and tumor unchanged in volume during the follow-up period. Three patients died of tumor progression, surgery and cancer after treatment 18, 22, 25 months respectively. Four patients underwent surgery respectively at 3, 4, 29 and 48 months after gamma knife operation. The local control rate of the tumors at 1 year was 92%, 2 years 88%, 3 years 80% and 4 years 75%. Pathological findings in these patients showed varying degrees of small vessel thickening and occlusion together with degeneration, necrosis in the center of tumor and loss of tumor cells at periphery.

CONCLUSIONSGamma knife is not adequately reliable for the control of hemangioblastoma cysts, it is an effective treatment of small or medium-size solid tumors, but long-term follow-up is needed. The recommended dose is 16 to 20 Gy.

Adolescent ; Adult ; Brain Neoplasms ; surgery ; Female ; Follow-Up Studies ; Hemangioblastoma ; surgery ; Humans ; Male ; Middle Aged ; Radiosurgery ; adverse effects ; methods ; Treatment Outcome ; Young Adult

Objective To explore the best methods and skill for the removal of difficult and high risk tracheobronchial foreign body under bronchoscope.Methods A retrospective review was performed between August 1995 to August 2012.There were 4217 children with tracheobronchial foreign body,among them,272 were diagnosed as high-risk,highly difficult tracheobronchial foreign bodies confirmed by clinical manifestations,foreign body type and bronchoscopy.Results In 271 children,the tracheobronchial foreign body was removed under bronchoscope,the success rate was 99.6％ ; only one child with a pen cap blocking the left lower lobe bronchus was transferred to the department of thoracic surgery,and the foreign body was finally removed by thoracotomy.Eighty-five children (among them,82 children were under 1 year of age)had Ⅱ-Ⅱ degree laryngeal obstruction,the emergency surgery was performed to remove the foreign body and to relieve the laryngeal obstruction.Twenty-six children had lung infection and 27 children had failed foreign body removal surgery before,in all these children,the foreign body was removed after infection control.There were 17 children with the pen cap as the tracheobronchial foreign body,direct removal was successful in 12 children with the history less than two weeks; in 4 children,the foreign body was removed after 0.1％ epinephrine saline flush,and 1 case with the homemade bronchial foreign body hook remove.There were 26 children with the whistle as the foreign body,and 32 children had large and sharp foreign bodies.In these cases,the foreign bodies were removed together with the bronchoscope.Forty-two children had multiple or fragile foreign bodies,and 16 children had subsegmental bronchial foreign bodies.In these cases,the foreign bodies were removed with forceps under direct vision and intraoperative bronchial lavage.In This series,129 children received intraoperative bronchial lavage,among them,127 children showed normal X-ray changes one week after operation.Two children with a history of more than 1 month complicated with pulmonary consolidation.After bronchial lavage,pneumothorax and subcutaneous emphysema occured,which recovered after treatment.No glottic edema,asphyxia,and other complications were found,the complication rate of surgery was 0.7％.Conclusion For the removal of highly difficult and high risk tracheobronchial foreign bodies,preoperative analysis and discussion shoule be sufficient,appropriate surgical skill and surgical instruments may improve the success rate of the surgery and prevent the operation complications.

OBJECTIVETo investigate the effects of heparanase expression inhibition on the proliferation, invasiveness and apoptosis of human lung adenocarcinoma cell line A549 cells.

METHODSRecombinant eukaryotic expression plasmid pshRNA-Hpa targeting human heparanase gene was constructed. A549 cells were cultured in DMEM and transfected with pshRNA-Hpa. The expression of heparanase mRNA and protein were examined by RT-PCR and Western blot. The proliferation, invasiveness and apoptotic rates of A549 cells were determined by MTT method, matrigel invasion assays and flow cytometry respectively.

RESULTSThe expression levels of heparanase mRNA and protein were down-regulated in A549 transfected with pshRNA-Hpa. The number of cells penetrating matrigel and the proliferation ability of A549 cells transfected with pshRNA-Hpa were reduced significantly compared to the control cells. The apoptotic rate of A549 cells transfected with pshRNA-Hpa was 12.53% +/- 0.34%, being significantly higher than that of the control cells (both P < 0.01). Western-blot showed that inhibition of heparanase expression led to reduced Akt phosphorylation.

CONCLUSIONSThe recombinant plasmid pshRNA-Hpa effectively inhibited the expression of heparanase, thus suppressing the proliferation and invasion and inducing apoptosis of A549 cells. The effects may be due to the down-regulation of Akt phosphorylation level.

Adenocarcinoma ; pathology ; Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Glucuronidase ; antagonists & inhibitors ; metabolism ; Humans ; Lung Neoplasms ; enzymology ; pathology ; RNA Interference ; immunology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; pharmacology ; Transfection