[ABSTRACT]OBJECTIVETo investigate the expressions of Endoglin (CD105) and Galectin-3 protein in laryngeal squamous cell carcinoma (LSCC) as well as the relationship between their expressions and the clinicopathological factors of LSCC.METHODSThe expressions of CD105 and Galectin-3 protein were detected in 76 samples of LSCC and 25 normal laryngeal tissues (NLT) by immunohistochemical staining (S-P).RESULTS 1.The mean of Microvessel density (MVD) value marked by CD105 in LSCC was 10.33±2.29, which was significantly higher than that in NLT (1.20±1.04) (P<0.05). The expression of MVD marked by CD105 (CD105-MVD) was correlated with histological grading, T stage, clinical stage, lymph node metastasis, recurrence and prognosis in LSCC (P<0.05). 2.The positive expression rate of Galectin-3 protein in LSCC was 86.84%, which was significantly higher than that in NLT (36%)(P<0.05). The expression of Galectin-3 was correlated with T stage,clinical stage, lymph node metastasis, recurrence and prognosis in LSCC (P<0.05). 3.There was a positive correlation between CD105 and Galectin-3 protein. 4.Survival analysis indicated that the expressions of CD105 and Galectin-3, histological grading, lymph node metastasis,T stage and recurrence were independent factors for tumor prognosis in LSCC (P<0.05). CONCLUSIONThe expressions of CD105 and Galectin-3 protein have a positive correlation in LSCC. They may play important roles in the tumorigenesis, malignant progression and poor prognosis of LSCC. Combined detection of them may be great value in diagnosis and predicting prognosis of LSCC.

Objective:To construct tumor cell model by determination of the pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid expressing steadily in mouse melanoma B16 cells.Methods:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid being constructed from the melanoma-associated antigen A3 genes sourcing laryngocarcinoma in advance was translated into the mouse melanoma B16 cells under the mediation of lipofectamine,and the positive clones were detected with G418.The expression of enhanced green fluorescent protein( EGFP) and MAGE-A3 mRNA in positive clones were detected by fluorescence microscopy and fluorescence quantitative PCR ( qRT-PCR ) assay, respectively.Results:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid has been transfected into B16 cells successfully, the green fluorescence of fusion protein expression was found, and MAGE-A3 mRNA transcription in B16 cells expressions were detected in positive clones.Conclusion:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid has been transfected effectively and expressed stably by liposome method in the B16 cells.The expression of MAGE-A3 tumor cell model has been successfully established,which provide data for the study of laryngocarcinoma immunotherapy.

OBJECTIVE: To provide a reference about choosing the methods of isolating exosomes derived from tumor cells including laryngocarcinima Hep-2 cells by comparing advantages and defects of two methods of isolation and extraction exosomes. METHOD: Previously, laryngocarcinoma Hep-2 cells were cultivated massively, then the cells were processed with hot shock in 42 degrees C for 1 h. Sucrose density gradient centrifugation ultrafiltration (method 1): cells culture supernatant 90 ml was gathered, the supernatant was clarified through a 3/0.8 μm small filter to remove impurities and fragments which in larger diameter. Then the filtering fluid was concentrated and purified through sucrose density gradient centrifugation and ultrafiltration, the concentrated fluid was obtained. Exosome Isolation Kit (method 2): cells culture supernatant 4 ml was gathered, the solutions of the kit were added into the supernatant in proper sequence, then filtered by the special column, the concentrated fluid was obtained. Both products are observed by high resolution transmission electron microscopy. RESULT: Both methods could isolate and extract exosomes feasibly. In single high power view of transmission electron microscopy, exosomes of method 1 disperse better, but lower density, and more impurity in background, exosomes of method 2 arrange closer, higher density, and less impurity. CONCLUSION Exosome isolation Kit require less supernatant, cost less time, process procedure briefly, harvest higher yield. It may become a new option of isolating exosomes derived from Laryngocarcinoma Hep-2 cells.
Cell Line, Tumor ; Exosomes ; ultrastructure ; Humans ; Laryngeal Neoplasms ; pathology ; Microscopy, Electron, Transmission

Objective: To explore the cytopathologic features and differential diagnostic essentials of the lung mucosa-associated lymphoid tumor(MALT).Methods: The fine needle aspiration(FNA) tissues and bronchi smears of 4 cases of MALT were examined by cytology,histology and immunohistochemistry respectively.Results: Cytologically,3 of the 4 cases were definitely diagnosed as lymphoid tumor and 1 suspected of small cell undifferentiated carcinoma,while histopathologically,all were diagnosed as MALT.The neoplastic cells were positive for CD20 and CD79a,but negative for CD3,CD5,CD10,CD45RO,CKpan and EMA.Conclusion: MALT is a rare pulmonary lymphoma difficult to be diagnosed.The definite diagnosis of pulmonary MALT depends on the cytology of FNA and bronchi smears,histopathology,and immunohistochemistry.

Objective:To investigate the clinic-cytology and histopathology of non-Hodgkin lymphoma (NHL) in breasts. Mothods:Twelve cases of NHL of the breast were studied by fine-needle aspiration biopsy (FNAC), histopathology and immunohistochemistry, and whose clinical data were analysed at the same time. Results:In 12 cases of NHL, 3 cases were T-cell NHL and 9 cases were B-cell NHL. Cytologically, the T-cell NHL cells were mostly arranging in diffuse patterns. The tumor cells were oval and pleomorphism. Some of them had distorted nucleus and thin nuclear envelope.The nucleus showed irregular course chromatin and visible nueleoic. Histopathologically, some of the tumor cells distributed around the blood vessels, and there was an obvious phenomenon of “blood-vessel-closing”. B-cell lymphoma cells were arranginy in diffuse pattern, and showed round and ellipse in shape with a clot and course granular chromatin and visible nueleoic and karyoknesis. Lymphoepithelia lesions were seen. Immunohistochemistry showed that CD3, CD43, and CD 45RO in T-cells NHL were positive.CD20, CD74 and CD79a in B-cells NHL were negative. CK, EMA, ER and PR in NHL were all negative. Conclusion:NHL of breast is extremely rare, and its definite diagnosis depends on various examination methods.

Objective: To investigate the advantage of ThinPrep cytological test in tumor diagnosis by fine needle aspiration cytology. Methods: Using fine needle aspiration for smears,some of smears were made in conventional smears,the remains aspirating needles were washed with a transport washing solution.ThinPrep slides were made using the ThinPrep Processor. Results: In 84 cases FNA samples,49 cases were breast,29 cases were lymph node and 6 cases were others.Comparing with conventionally prepared specimens,ThinPrep processed samples showed the advantage in sensitivity or specificity.The lack of cellular debris,blood and exudates improve the ability to detect cellular abnormalities and speed up the time required for screening in ThinPrep processed samples.Rapid cell transfer from the sampling device to the transporing fluid containing the fixative ensure preservation of cell details with minimal artifactual changes. Conclusion: The use of Thinprep and by combination of FNAC and immunocytochemistry cytopathological diagnosis can be improved significantly.

Objective To understand the changes of allergic rhinitis from immunology,pathophysiology and morphology method were applied.Methods Toluene 2,4-diisocyanate was dissolved with florence oil to final concentration 10%,this solution was dropped into the nasal vestibules of animals to induce sensitization process.8 guinea pigs were prepared as allergic rhinitis with 10% Toluene 2,4-diisocyanate,and other 8 animals were used as blank controh.After model successfully established,the blood were obtained to determine RBC-C3b receptor rosette rate and RBC-imuuunocomplex rosette rate.The nasal mucosas were obtained from 2 groups,distribution and changes of substance P in nasal mucosa were observed with the application of immunohistochemical staining.The content of blood histamine was determined with ELASA method.The pathological changes of nasal mucosas were observed with the application of light microscope and transmission electron microscope.Results The behavior scores of the modeling animals were significantly higher than that of controls(P＜0.01).The value of RBC-C3b reeeptor rosette rate and RBC-imuuunoeomplex rosette rate of the modeling animals were significantly decreased than that of the controls(P＜0.05).Substance P presented in the normal nasal mucosal epithelial cells,epithelium cells of blood vessels,glandular cells and its duets,the staining density and the positive staining cells in the same aero in modeling group significantly increased,compared with controls.The counts of substance P-positive cells of control group were less than those of modeling group(P＜0.05).The content of blood histamine of the modeling animals were significantly increased compared with the controls(P＜0.01).The structure of false multiple coat cilium columnar epithelial cells in nasal mucosa of control group were successive,intact,and distinct.There were normal mucosal epithelium,lamina propria and submucosa.But modeling group showed that mucosal epithalamiums were damaged and shed,goblet cell proliferated,squamous metaphase,and epithelial necrosis happened,serous glands in lamina propria vigorously proliferated,blood vessels expanded,tissue edema formed,plenty of inflammatory cell such as eosinophil and mast cells were more in number and infiltrated.The structure of epithelial eells,cilia and mierotubule of nasal mucosa of control group were regular and distinct,there were abundant cellular organs in cytoplasm.Eosinophil cells were intact.But iu model group,mucosal epithalamium,cilia and its microtubule,mierovillus,goblet cell were damaged,the cell bulk and nucleus changed,mast cells and eosinophil cells changed,blood vessels expanded,serous glands vigorously proliferated.This morphological change was roughly identical to clinical manifestation of allergic rhinitis.Conclusion Toluene-2,4-diisocyanate can be used to establish allergic rhinitis model in guinea pigs,and some ehanges of the allergic rhinitis in model guinea pig were similar with clinic observation.

AIM: To observe the effect of partial liquid ventilation with perfluorocarbon on gas exchange, hemodynamics and lung histological change in the piglets with surfactant depletion-induced acute lung injury. METHODS: 12 piglets were treated by lung lavage to a partial pressure of oxygen in arterial blood (PaO 2) below 100 mmHg for one hour and randomly divided into gas ventilation group (control group) and partial liquid ventilation (PLV) group, in PLV group, piglets received PFC (FC3280) intratracheally at doses of 15 mL/kg. The parameters of gas exchange and hemodynamics were measured before lung lavage, after lung lavage when the acute lung injury (ALI) was established, and 1 hour, 2 hours after ALI. Animals underwent euthanasia at the end of the study, lung histologic analysis followed. RESULTS: Surfactant depletion by lung lavage induced a stable acute lung injury.Gas exchange increased markedly in the animals that underwent PLV, less hemodynamic damage was observed in PLV group compared with the animals in GV group. Lung histologic analysis demonstrated a less lung damage, including atelectasis, neutrophil excudation, intra-alveolar hemorrhage and interstitial edema in PLV group compared with control group. CONCLUSION: In piglets with surfactant depletion-induced acute lung injury, partial liquid ventilation with perfluorocarbon can improve the gas exchange with less adverse hemodynamic effect and less lung injury compared with conventional gas ventilation.

Objective To understand the human resource status of the community rehabilitation management in Shanghai and provide evidence for the construction of this human resource. Methods Census institutions of community rehabilitation management in Shanghai were investigated with self-questionnaire. Results Shanghai has initially established personnel of community rehabilitation management,which contains rehabilitation coordinators and street community administrator. However, the structure and number of the personnel has yet to be improved. Conclusion Personnel building must focused on optimizing the personnel structure of rehabilitation coordinator, moderately expanding the street community administrators, filling the community rehabilitation service gap between city and urban in next stage.

ObjectiveTo analyze past study on senile dementia in China, and shape the foundation for constructing social support system for elderly with dementia.MethodsA combination of qualitative and quantitative method was used to analyze 406 literatures related to the elderly with dementia published in China since 2000.ResultsDiagnosis method for the senile dementia has matured; but the causes of the disease is still not clear and there is no thorough remedy; study on early intervention, nursing care and social support are inadequate.ConclusionDiagnostic criteria should be standardized, community screening assessment should be done on a regular basis; etiology based on risk factors should be strengthened; systematic study on intervention should be carried out; training on home care techniques should be enhanced. Resolving the care of the elderly with dementia depends on the social support system construction eventually.