Objective To study the relation of the porin at the out membrane with resistance to cefoxitin in Enterobacter cloaceae. Methods The extended spectrum beta lactamase(ESBLs) was detected by phenotypic confirmatory test according to NCCLS M100 S9;the beta lactamase and out membrane protein(OMP) was extracted by ultrasonication,the beta lactamase was detected by ultraviolet spectrophotometer,the composition of OMP was analyze by SDS PAGE. Results The 6 strains in 9 strains of Enterobacter cloaceae with resistance to cefoxitin lost 18 kda protein band and did not product hydrolase to cefoxitin and 5 strains was ESBLs positive. 2 strains producing hydrolase to cefoxitin did not lose the 18 kda protein. One strain with resistance to cefoxitin was producing ESBLs but did not defect the 18kda protein band and product hydrolase to cefoxitin. Conclusion The defect of the 18 kda protein band is nearly relation with resistance to cefoxitine in Enterobacter cloaceae.

Objective To study a new clinicopathological subtype of primary cutaneous T-cell lym phoma (PCTCL). Methods A case of T-cell lymphoma was systematically evaluated clinically and by using H-E staining, special staining,immunohistochemical staining,gene rearrangement and PCR.Results The skin lesion presented as tender nodules with mucocele. Skin biopsies showed that in the dermis and subcutaneous tissue,most of the angiotropic tumor cells were small T cells; no obvious epidermotropic phenomenon was detected.A few vessels were observed with obvious fibromucinous matrix formation. Immunohislochemical studies showed the following:CD3(+),CD43(+),CD45RO(+),CD56(a few),CD68(-), CD79?(-),CD20 (-), CD30(-), CD117(-), ALK(-), S-100(-),CD45R(-),EMA(-),SMA(-).The mucoid matrix was positive for Alcian blue staining.The rearrangement of T-cell ? receptor gene was detected.EBV was not detected with PCR.Conclusion Fibromucinous T-cell lymphoma rich in blood vessels is a new and distinct variant of PCTCL; it is not a subtype of mycosis fungoides.

Objective To investigate the imaging diagnosis of vocal cord paralysis.Methods Of 58 patients,53 cases was unilateral and 5 bilateral.All patients underwent laryngeal tomography and 6 of them underwent CT.The imaging findings were analyzed.Results The tomography findings of unilateral vocal cord paralysis were as follows:dilated piriform sinus,41.5%;asymmetrical vestibule,39.6%;enlarged ventricle,71.7%;thinning cord,47.2%;paralyzed cord higher than intact cord,73.6%;cord fixed in adducent position during inspiration,100%;tilting of the thyroid cartilage,11.3%.The cord fixed in paramedian was the main appearance of bilateral vocal cord paralysis.The CT features of vocal cord paralysis included dilated piriform sinus(6 cases),enlarged laryngeal ventricle(4 cases),fixed cord in paramedian position(4 cases)and tilted anteromedially arytenoid cartilage(5 cases).Conclusion To evaluate the functional abnormality of larynx,tomography is useful and CT is well to display the laryngeal anatomic structures and changes,therefore,if the diagnosis is uncertain by tomography or the intrinsic lesions of larynx is suspected,CT is recommended.

OBJECTIVETo study the effects of RhoA down-regulation by RNA interference on the invasion of tongue carcinoma Tca8113 and SCC-4.

METHODSDetermination of the human RhoA sequence as well as the design and constructionof a short specific small interfering RNAs (siRNA) were performed. The siRNA of RhoA gene was transfected into humantongue squamous cell carcinoma Tca8113 and SCC-4 cells line by Lipofectamine 2000. Quantitative real-time polymerasechain reaction was used to examine the mRNA expressionlevels of RhoA. Protein expressions of mRNA, galectin-3,and matrix metalloproteinase (MMP)-9 were evaluated byWestern blot. Transwell invasion assay was performed toassess the invasion ability of tongue carcinoma.

RESULTSRhoA expressions in Tca8113 and SCC-4 cells were reducedsignificantly after transfection of RhoA-siRNA. Protein levels f galectin-3 and MVP-9 were also down-regulated significantly. Invasion ability was inhibited as well.

CONCLUSIONRhoA-siRNA can effectively inhibit RhoA expression in Tca8113 and SCC-4 cells. The invasion ability of tongue carcinoma cells decreased with down-regulation of the protein expressions of galectin-3 and MMP-9, indicating that RhoA-siRNA can inhibit invasion of tongue carcinoma. Results show that RhoA may play an important role in the processes of invasion and metastasis of tongue carcinoma.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Galectin 3 ; metabolism ; Gene Silencing ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Tongue Neoplasms ; genetics ; metabolism ; pathology ; Transfection

Protein A and protein G are two well-defined immunoglobulin (Ig)-binding proteins (IBPs), which show affinity for specific sites on Ig of mammalian hosts. Protein A and protein G contained several highly homologous IgG-binding domains which had been demonstrated to have function to bind to IgG. Whether combinations of Ig-binding domains of various IBPs could produce useful novel binding properties remains interesting. We constructed a combinatorial phage library which displayed randomly-rearranged A, B, C, D and E domains of protein A, B2 and B3 domains of protein G. Four rounds molecular evolution of this library directed by all four human IgG subclasses respectively generated a common arrangement of D-C respectively which didn't exist in SpA. The dynamic loss of control phages and increase of the phages displaying two or more binding domains, especially the selective enrichment of D-C and strict selection of its linking peptides demonstrated the efficient molecular evolutions and the significance of the selected D-C arrangement. The phage binding assays confirmed that D-C possessed a binding advantage with four human IgG subclasses compared to SpA. In this work, a novel combination of Ig-binding domains, D-C, was obtained and presented the novel Ig binding properties which provided a novel candidate molecule for the purification, production and detection of IgG antibodies and a new approach for the further study of structures and functions of IBPs.
Amino Acid Sequence ; Antibody Specificity ; Bacterial Proteins ; immunology ; metabolism ; Binding Sites ; Binding, Competitive ; Evolution, Molecular ; Immunoglobulin G ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Sequence Alignment ; Staphylococcal Protein A ; immunology ; metabolism

We constructed a phage-displayed random mutation library of Tat38-61(51N/55N), for studying the molecular evolution screening of HIV-1 Tat38-61 epitope. We used primers containing the random nucleotide sequences, and introduced the random mutations at the sites of 51 and 55 amino acids coding sequences into full-length Tat sequences by overlapping PCR. With the randomly mutated full-length Tat as template, the Tat38-61(51N/55N) mutants which contained recognition sequences for the Xba I in both ends were amplified by PCR using the designed primers. The mutants were cloned into Xba I site in the phagemid vector pCANTAB5S, then the recombinants were transformed into E. coli TG1, a phage-displayed the random mutation library of Tat38-61(51N/55N) was constructed by the rescue of help virus M13KO7. The results showed that the library consisted of about 5.0 x 10(6) colonies and the phage library titer was 2.65 x 10(12) TU/mL. More than 56.50% colonies in the library were positive for insertion. Sequence analysis showed that the nucleotides encoding amino acids at the sites of 51 and 55 distributed randomly. The constructed mutation library could meet the requirements for the following molecular evolution screening, and might prepare the Tat mutants for the further study of new Tat vaccine candidates.
AIDS Vaccines ; immunology ; Escherichia coli ; genetics ; metabolism ; HIV-1 ; genetics ; Humans ; Mutation ; Peptide Fragments ; biosynthesis ; genetics ; immunology ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; tat Gene Products, Human Immunodeficiency Virus ; biosynthesis ; genetics ; immunology