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PURPOSE: To explore the reconstructive methods by examining and evaluating the rehabilitation of swallowing function with videofluoroscopic swallowing study (VFSS). METHODS: The patients were divided into 3 group, including control group (healthy volunteers, n=10), mobile tongue group after reconstruction (n=10), and tongue base group after reconstruction (n=10). The dynamic swallowing process was recorded by VFSS in all subjects. The data in 3 groups included the performance of dysphagia (irritating cough or aspiration) and the incidence rate of irritating cough and aspiration, the change of the ability to swallow viscous food, the change of residual amount (oral cavity and pharyngeal space), the change of pharyngeal clearance time. The results were analyzed with Kruskal-Wallis test by SASS14.0 software package to compare whether there was significant difference in the swallowing function among 3 groups. RESULTS: The risk of postoperative irritating cough and aspiration was not increased in the patients with defect in mobile tongue, but significantly increased in patients with defect in tongue base(0%,0%,60%). The patients ability to swallow viscous food in control group (9.33±0.50 mL) was better than that of tongue mobile group (6.25±2.12 mL) and tongue base group (0.70± 1.16 mL). Swallowing efficiency in control group was better than that of mobile tongue group and tongue base group, and the latter was the worst. CONCLUSIONS: The decrease in swallowing efficiency in tongue base group is greater than that of mobile tongue group, the choice of reconstructive method depends on site and volume of the tongue defect. Supported by Research Fund of Science and Technology Commission of Shanghai Municipality (08DZ2271100) and Key Project of Biomedicine of Science and Technology Commission of Shanghai Municipality(10DZ1951300). [ABSTRACT FROM AUTHOR]

Hepatitis B virus (HBV) is a major cause of chronic liver disease, and frequently results in hepatitis, cirrhosis, and ultimately hepatocellular carcinoma. HBV polymerase (Pol) is an essential viral protein that is important for HBV replication and might be involved in the development of hepatocellular carcinoma. Protein-protein interactions appears to be crucial for its role. The aim of this study was to screen and identify the proteins that interact with Pol using a co-immunoprecipitation-based LC-MS/MS identification technique. The HBV Pol gene was amplified by polymerase chain reaction (PCR) and cloned into pCDNA3.1(+). The recombinant plasmid pCDNA3. 1(+)-Pol-flag was transfected into HeLa cells. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) identified 45 proteins that co-immunoprecipitated with flag-tagged HBV Pol. Eleven of these have previously been reported as proteins that interact with HBV Pol. A proof-of-concept-based Ingenuity Pathway Analysis (IPA, www.ingenuity.com) was used to characterize the functions and pathways of these 45 identified proteins and HBV Pol. Among these proteins, four proteins may play a role in three major molecular cellular networks, and are therefore worthy of further investigation.

Objective: To investigate the function of hepatitis B virus polymerase (HBV Pol) in the viral life cycle by screening the proteins interacting with HBV polymerase., Methods: The HBV Pol gene was constructed into the pGBKT7 vector. GAL4 yeast two-hybrid system was used to screen the human liver cDNA library to obtain proteins which interacted with HBV Pol. GST-pull down assay was applied to confirm the protein interactions., Results: Ubiquitously expressed transcript (UXT) was selected by the yeast two-hybrid system. GST-pull down assay confirmed the in vitro interaction between HBV Pol and UXT., Conclusions: UXT is a potential interactor of HBV Pol, and this protein interaction may provide clues of the function of HBV Pol in HBV life cycle.

Aim: To construct the recombinant plasmid of HCV core protein, and to express and identify it in normal human hepatocyte HL-7702., Methods: HCV core gene was cloned by using PCR from plasmid pBRTM/HCV1-3011 which included the full length of HCV gene. The core segment with expression plasmid pcDNA3.1(-) was recombined to construct eukaryotic expression plasmid pcDNA3.1(-)/core, which was then transfected into human hepatocytes by using poly-cation. The expression of core protein was detected by immunochemical staining and Western blot., Results: The length and sequence of the cloned core segment were correct. The transfected HL-7702 cells expressed the core protein., Conclusion: The eukaryotic expression plasmid pcDNA3.1(-)/core including HCV core gene is successfully constructed. The effective expression of HCV core protein in human hepatocytes is useful for further development of HCV core antigen.

Objective: To investigate the effect of cell surface sialic acid and its linkage on the cell-cell and cell-matrix adhesion of mammary carcinoma cells MD-MB-435., Methods: MD-MB-435 cells were sense-transfected with ST6Gal I cDNA or antisense-transfected with part of the ST6Gal I sequence inserted in pcDNA 3.1 vector, with mock transfection with pcDNA3.1 vector as the control. The cell surface alpha2, 6-linked sialylation was determined by fluorescence-activated cell sorting (FACS) using lectin SNA (Sambucus nigra agglutinin specific to alpha2, 6-linked sialic acid on N-linked glycoprotein). A significantly increased alpha2, 6-sialylation subclone in sense-transfectants and a decreased alpha2, 6-sialylation subclone in antisense-transfectants were selected for further examination of cell-cell and cell-matrix (collagen IV) adhesion. The transfectants were also treated with sialidase to compare the capacity of cell adhesion affected by cell surface sialylation., Results: Sense-transfection subclone showed a reduced cell-cell aggregation but enhanced cell-matrix adhesion. In contrast, the antisense-transfection subclone exhibited increased cell-cell aggregation and decreased cell-matrix adhesion. After treatment with sialidase, the cell-matrix adhesion of all the transfectants and the parental MDA-MB-435 cells were significantly reduced to the level of 31%-57% of untreated cells., Conclusion: Cell surface sialic acid and alpha2, 6-linked sialylation play an important role in cell-cell and cell-matrix adhesion of mammary carcinoma cell MDA-MB-435.