Objective To explore a fast and precise registration algorithm for megavolt (MV) portal images(PIs) used for radiotherapy positioning verification, and find auto analysis method of set-up error using the computed image processing and mutual information comparison technology, which provide a basis for the development of automatic image guidance software. Methods MV PIs of patients undergoing radiotherapy were tested, pre-processed with noise reduction technique based on improved filtering algorithm and contrasted by gray-scale transforming using partial derivative threshold. The bone structures were then highlighted but soft tissues and the cavities were restrained simultaneously. Improved particle swarm optimization and powell hybrid algorithm were used to optimize and transform the mutual information based on wavelet multiresolution analysis when registering the Pls with digital reconstructed radiographs (DRRs) of treatment planning or X-ray simulation-film images(SIs). Application of the designed registration algorithm was verified and evaluated through simulated set-up shifts of head and neck phantom. Results The improved noise reduction algorithm satisfactorily met the requirements for contrast of bony structures in the MV PIs. The established mutual information registration method well behaved in both accuracy and speed of registration calculation. The processing of automatic registration took only 31.4 seconds averagely for the PIs and X-ray Sis of head-neck phantom. Mean errors of automatic registration of PIs and X-ray Sis in horizontal, vertical and rotational reduced by 62. 74% ,67. 32% and 66. 61% respectively compared with manual registration in the testing of 20-cases head and neck phantom. Conclusions A precise image registration algorithm and set-up error analysis method based on MV portal images is established, and it can meet the clinical application in registration accuracy and speed.

OBJECTIVETo observe effect of oral scorpio and scolopendra powder on T-cell subsets in peripheral blood and intestine from rats with collagen induced arthritis (CIA).

METHOD60 rats were randomly divided into 6 groups: normal control group, model control group, low-dose scorpio and scolopendra group, middle-dose scorpio and scolopendra group, high-dose scorpio and scolopendra group, and type II collagen group. Rat's rheumatoid arthritis was induced by collagen II (C II). Level of T-cell subsets from peripheral blood and intestine was measured by flow cytometry.

RESULTCD4+ T cellular level was obviously increased (P < 0.05 or P < 0.01) or kept increased tendency in peripheral blood and intestine from the model group compared with that of the normal group, while the ratio of CD4+/CD8+ in intestine was obviously descent but the contrary in peripheral blood (P < 0.05 or P < 0.01). CD4+, CD8+ T cellular level in intestine were obviously descent and the ratio of CD4+ /CD8+ increased in all treated groups when compared with in the model group (P < 0.05 or P < 0.01). However, CD4+ T cellular level and the ratio of CD4+/CD8+ in peripheral blood were remarkablely decreased.

CONCLUSIONThe mechanism that scorpio and scolopendra could treat rat's rheumatoid arthritis may be regulating balance of T-lymphocyte subsets in peripheral blood and intestine.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Arthritis, Experimental ; immunology ; Arthritis, Rheumatoid ; immunology ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; Female ; Intestinal Mucosa ; immunology ; Medicine, Chinese Traditional ; Rats ; Rats, Wistar ; Scorpions ; chemistry ; T-Lymphocyte Subsets ; drug effects ; immunology

OBJECTIVETo develop a recombinant single domain antibody against hTERT, human telomerase catalytic subunit.

METHODSA previously prepared His-tagged hTERT fusion protein was used as the antigen, and the variable regions in heavy chain (VH) of immunized mice were RT-PCR amplified and cloned into the pCANTAB 5E, a phagemid vector. By transfection, the display library of mouse VH was developed. The candidate clones were selected by affinity panning, and soluble VH were obtained after expression in E. coli, HB2151. The resultant single VH antibodies were characterized on their binding potentials by western blotting.

RESULTSAn about 350 bp VH fragment was amplified from spleen cells of mice immunized by His-tagged hTERT and expressed by phage displayed as VH library. The size of the library was 8 x 10(4). After three rounds of affinity panning, 4 independent clones were chosen and consequently expressed as soluble single domain antibodies (Mr = 16 000). In Western blot analysis, the single domain antibody from 2 of 4 clones proved to react with the His-tagged hTERT fusion protein (Mr = 167 000) without dependence of His-tags and also detect the native hTERT (Mr = 127 000) extracted from the human HeLa cancer cell line. DNA sequencing showed both of the single domain antibodies were encoded by the heavy chain variable region of the mouse.

CONCLUSIONSThe single domain antibodies developed were hTERT recognizable and hTERT specific, thus providing a basis for application of recombinant single domain antibody in inhibition of telomerase activity and anticancer therapy.

Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; genetics ; immunology ; Base Sequence ; Cloning, Molecular ; Complementarity Determining Regions ; genetics ; DNA-Binding Proteins ; HeLa Cells ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Mice ; Molecular Sequence Data ; Sequence Analysis, DNA ; Telomerase ; immunology

OBJECTIVETo evaluate therapeutic effect of Sishen Wan on experimental colitis, and explore its mechanism by expression of Bax/Bcl-2 mRNA, Fas/FasL in colonic tissue.

METHODExperimental colitis was induced by rectal administration of trinitrobenzene sulfonic acid (TNBS) dissolved in ethanol. The model animals were divided into four groups: the induced colitis but untreated group, the induced colitis groups treated with the high, middle, low dose of Sishen Wan, and the induced colitis group treated with salicylazosulfapyridine (SASP). After 10 day administration, the body weight, colonic wet weight, colonic weight index, colonic damage score and pathological change were evaluated, and the level of Fas and FasL by flow cytometry, Bax mRNA and Bcl-2 mRNA by reverse transcription polymerase chain reaction (RT-PCT).

RESULTCompared with the model group, the colonic wet weight and colonic weight index were remarkably decreased in the middle dose of Sishen Wan group (P < 0.05). The colonic injury scores were significantly reduced after rats were treated with the three doses of Sishen Wan (P < 0.05). Representative restored features were observed including fewer inflammatory cellular infiltration and follicular hyperplasia, superficial and little ulcer with fibroplasia in colonic mucosa from the treated groups. The expression of Fas in the colonic mucosa was obviously down-regulated (P < 0.05) and the ratio of Bcl-2 mRNA/Bax mRNA was significantly up-regulated (P < 0.05) in the groups treated with the three doses of Sishen Wan.

CONCLUSIONSishen Wan might postpone colonic epithelium apoptosis or improve inflammatory cell apoptosis by regulating the expression of Fas/ FasL and Bax/Bcl-2 mRNA in colonic tissue, which is possible potential path to effectively treat experimental colitis by enema.

Animals ; Colitis ; drug therapy ; metabolism ; Colon ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fas Ligand Protein ; genetics ; Female ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; genetics ; fas Receptor ; genetics