ABSTRACT
Objective To investigate the clinical characteristics on diagnosis and treatment of renal tubercu- losis.Methods Clinical data of 82 patients with renal tuberculosis were retrospectively reviewed.Results All of the 18 cases who received medication recovered completely.64 cases undergoing surgery were pathologically diagnosed to have renal tuberculosis,of which 2 cases developed ureteral stump syndrome.Conclusion Urine PCR-TB-DNA re- mained the primary diagnostic method before operation.Computed tomography(CT)and magnetic resonance urogra- phy(MRU)could be used in diagnosis of renal tuberculosis.When the non-functioning kidney was resected,per- inephrit fat and the involved ureter should be concomitantly resected as much as possible.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the mechanism of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells.</p><p><b>METHODS</b>In this study, the proteins extracted from paclitaxel-induced apoptotic MCF-7 cells were analyzed by 2-dimentional gel electrophoresis (2-DE), and compared with those from untreated MCF-7 cells. The differential proteins were identified by mass spectrometry.</p><p><b>RESULTS</b>At 24 hour after paclitaxel (100 nmol/L) treatment, MCF-7 cells were collected and extracted the whole proteins. Seventeen up-regulated or down-regulated proteins were found by analysis of the differential proteomic 2-DE map. Six of them were identified by mass spectrometry. They were enolase 1, chloride intracellular channel 1, keratin 8, ribosomal protein S12, galectin-1 and histidine triad nucleotide binding protein, respectively.</p><p><b>CONCLUSION</b>We effectively found the changed proteins in the process of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells by proteomic techniques. These up-regulated or down-regulated proteins are important molecules for our further research about the mechanism of paclitaxel-induced apoptosis.</p>
Subject(s)
Female , Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Breast Neoplasms , Metabolism , Pathology , Cell Line, Tumor , Electrophoresis, Gel, Two-Dimensional , Galectin 1 , Metabolism , Keratin-8 , Metabolism , Paclitaxel , Pharmacology , Phosphopyruvate Hydratase , Metabolism , Proteomics , Methods , Ribosomal Proteins , MetabolismABSTRACT
The aim of this study is to construct a phage display single-chain variable fragment (scFv) library against breast cancer cells and screen the specific antibodies against MCF-7 cells from the library. The BALB/C mice were immunized with MCF-7 cells. Total RNA of spleens was isolated. The heavy-chain (VH) and light-chain variable region genes (VL) of the antibodies were amplified by RT-PCR and joined into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The assembled scFv fragments were cloned into the phagemids(pCANTAB5E) and the recombinant phagemids were used to transform competent E. coli TG1. The transformed TG1 cells were infected by helper phage M13KO7 and the recombinant phagemids were rescued. The scFv fusion proteins were displayed on the surfaces of the recombinant phages. A phage display antibody library of repertoire of 1.2 x 10(6) clones was constructed. The specific antibodies against MCF-7 cells were enriched by 75 times after five rounds of affinity selection. Ten recombinant phages clones that exhibited specific binding to MCF-7 cells were identified. The specificity of those phage clones was analyzed by reactivity against HepG2 cells and Hela cells by ELISA. One of the selected phage clones against MCF-7cells was used to infect E. coli TOP10 to produce the soluble scFv antibodies after induction with IPTG. The strategy of construction and screening of antibody library directed against the whole tumor cells described in this report should be generally applicable to generate tumor cell-specific antibodies.