Objective To explore the effect of RNA interference on the bionomics of Cathepsin L of hepatoma carcinoma cells. Methods In this study, the experimental group was the Cathepsin L RNA interference group. Control groups were the normal blank hepatoma carcinoma cell group (the blank group) and the Cathepsin LRNA interference blank group (the fluorescence control group). Observing times were ld,3d and 6d after RNA interference. Transfection efficiency in each group was observed. Expression of Cathepsin L of hepatoma cells was detected by immunofluorescence, RT-PCR and WB. Cell vigor was detected by MTT assay. Changes in the cell cycle and apoptosis were observed by flow cytometry. Transwell cabin was used to detect the changes of cell invasive power.Results In the experimental group, Cathepsin L mRNA level and protein level significantly decreased, the proliferation index significantly decreased and apoptosis index significantly increased. The invasive power also decreased. Conclusion RNAi interference can inhibit Cathepsin L expression, cell proliferation and cell invasive power efficiently.

No abstract available.
Cyclosporine ; Necrobiosis Lipoidica ; Necrobiotic Disorders

No abstract available.
Dermatitis, Allergic Contact ; Petrolatum

Objective To investigate the molecular mechanism of epithelial-mesenchymal transition (EMT) induced by activated vascular endothelial growth factor receptor-1 ( VEGFR-I ) in cell line MHCC97-H.Methods MHCC97-H cells were cultured in DMEM with 1％ fetal bovine serum (control group),10 μmol/L PP2 (PP2 group),10 μmol/L PBS (PBS group),50 μmol/L VEGF-B (VEGF-B group),l0μmol/L PP2 and 50 μmol/L VEGF-B (PP2 +VEGF group),10 μmol/L PBS and 50 μmol/L VEGF-B (PBS + VEGF-B group),respectively.Protein expressions of epithelial marker E-cadherin,α-catenin and mesenchymal marker vimentin and N-cadherin were detected by Western blot.The expression sites of E-cadherin,α-catenin and mesenchymal marker vimentin and N-cadherin were detected by cell immunofluorescence.The ability of invasion and migration of cell line MHCC97-H were assessed by cell invasion and migration test.All data were analyzed by the t test.Results The expressions of E-cadherin,α-catenin,vimentin and N-cadherin were 3.23 +0.76,3.01 ±0.25,3.01 +0.22 and 2.63 +0.40 in the control group,4.18 +0.32,3.29 +0.11,4.85 +0.36 and 3.02 +0.52 in the PP2 group,2.83 +0.65,3.03 +0.27,1.37 ±0.24 and 2.98 ±0.36 in the PBS group,2.06 ±0.15,2.84 ±0.76,5.79 ± 0.38 and 5.54 ± 0.28 in the VEGF-B group,6.12 ± 0.08,5.45 ± 0.37,3.36 ± 0.42 and 3.26 ±0.13 in the PP2 + VEGF-B group and 1.36 ±0.54,1.26 ±0.45,4.05 ±0.17 and 1.05 ±0.33 in the PBS +VEGF-B group.There was a significant difference in the expressions of E-cadherin and α-catenin between the PP2 +VEGF-B group and the VEGF-B group (t =7.625,9.931,P ＜ 0.05 ).The expressions of vimentin and N-cadherin in the PP2 + VEGF-B group were significantly lower than those in the VEGF-B group (t =12.001,11.910,P ＜ 0.05).Six hours after the treatment with VEGF-B,the numbers of MHCC97-H migrated were 19 ± 1,5 ± 2and 16 ± 1 in the VEGF-B group,PP2 + VEGF-B group and PBS + VEGF-B group,respectively.The number of MHCC97-H cells migrated in the VEGF-B group was greater than that in the PP2 ± VEGF-B group ( t =13.566,P ＜ 0.05 ).The number of MHCC97-H cells passed through the Boyden chamber was 4 + 2,which was significantly less than 16 ± 1 of the VEGF-B group (t =12.350,P ＜0.05).Conclusion EMT induced by activated VEGFR-1 was mediated via c-Src kinase signal transduction in MHCC91-H cell line,and c-Src may be a potential target to interfere the invasion and migration of hepatic cancer cells.

No abstract available.
Erythema* ; Flushing* ; Rosacea*

No abstract available.
Interferon-gamma Release Tests* ; Lupus Vulgaris*

In order to investigate the cellular immunoresponses mediated by chimeric anti-CD20 monoclonal antibody (anti-CD20 McAb) through dendritic cells (DCs), mononuclear cells were isolated from human peripheral blood (PBMNC) and DCs from PBMNCs in vitro were generated with normal methods. Human CD20 positive lymphoma cell line-Raji cells were treated with different methods including treatment with anti-CD20 McAb (group R), treatment with heat-induced apoptosis (group A) and treatment with anti-CD20 McAb+heat-induced apoptosis (group R+A), then Raji cells treated with above-mentioned methods as tumor antigen were loaded on DCs. The phagocytosis of DCs to tumor antigen was observed by transmission electron microscope (TEM), the auto-mixed lymphocyte reaction was performed with antigen-primed DCs, the release of INF-gammafrom effector cells was detected by ELISPOT, the killing of effector cells on Raji cells was assayed by MTT. The results showed that under TEM, no pronounced phagocytosis of DCs was seen in group R, while the phagocytosis of apoptotic bodies could be easily seen in group A and the more cell fraqments were observed in group R+A. The FCM indicated that the expressions of CD80, CD86 and HLA-DR on DCs in 3 experimental groups were higher than those in group control (p<0.05), while expression positive rate in group R+A was higher than those in group R and A (p<0.05). The detection of lymphocyte surface antigen revealed that proportions of CD8+ cells in all experimental groups were higher than those in group control (p<0.05), count of CD56+ cells in group R and R+A increased, as compared with group A and control, difference was significant (p<0.05). ELISPOT assay indicated that amount of cells releasing IFN-gamma in all experimental groups was higher than that in group control, and also number of spots in group R+A significantly higher than that in other groups at effector-targetor ratio=1:10 (p<0.05). The results of killing assay demonstrated that killing rate on Raji cells in all experimental groups increased as compared with group control (p<0.05), while killing rate in group R+A was higher than that in group R and A. It is concluded that anti-CD20 McAb can mediate DC to induce cellular immunoresponse against lymphoma, that is, to stimulate and amplify specific CTLs and NK cells. Anti-CD20 McAb combined with DCs primed by heat-stressed tumor cells as antigen can further enhance cellular immunoresponse against lymphoma.
Animals ; Antibodies, Monoclonal ; pharmacology ; Antigen Presentation ; Antigen-Presenting Cells ; immunology ; Antigens, CD20 ; immunology ; Chimera ; Dendritic Cells ; immunology ; Humans ; Hyperthermia, Induced ; Lymphoma, B-Cell ; immunology ; Mice ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo develop a virulent heat-evil-induced thrombosis animal model, and provide a rational animal model for pathogeny and pathogenesis research of thrombosis-related diseases, anti-thrombosis activity screening and pre-clinical studies of CAHT formula.

METHODSD rats were pretreated with carrageenin (Ca) intraperitoneal injection, followed by intravenous injection of endotoxin (LPS from E. coli O111:B4) 50 microg x kg(-1) 16 h later. Thrombosis in rat tails were observed during 12-24 h after injection of LPS. The inflammatory mechanism of this model were investigated by analyzing serum level of TNF-alpha, IL-6, TXB2 and 6-keto-PGF 1alpha, CD11b/CD18 expression of white blood cells (WBC) and P-selectin expression of vessel walls.

RESULTIn LPS/Ca model group, thrombosis can be clearly observed in the distal part of rat tails after 12-24 h of LPS/Ca treatment. High level of TNF-alpha and IL-6 can be measured in serum. The expression of CD11b/CD18 in WBC and P-selectin in vessel endothelium significantly increased and the number of WBC in peripheral blood markedly decreased shortly after LPS/Ca treatment. The adherence of white blood cells to vessel endothelium which can be seen by microscope mainly contributed to the decrease of WBC. The results indicated that there was obvious inflammation after treatment with LPS/Ca, suggesting that inflammation was the key mechanism for this model.

CONCLUSIONThis model was developed through treatment of LPS in combination with Ca, of which LPS is considered to be an exotic virulent heat-evil in TCM, while the inflammatory molecules produced in this model, such as TNF-alpha, IL-6, CD11b/CD18 and P-selectin belong to internal virulent heat-evils, so this animal model consists of pathogeny and pathogenesis of virulent heat-evils. virulent heat-evil.

6-Ketoprostaglandin F1 alpha ; blood ; Animals ; CD11b Antigen ; metabolism ; CD18 Antigens ; metabolism ; Carrageenan ; pharmacology ; Disease Models, Animal ; Endotoxins ; pharmacology ; Immunohistochemistry ; Interleukin-6 ; blood ; Leukocytes ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Thrombosis ; blood ; chemically induced ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo investigate interleukin-1 receptor antagonist (IL-1ra) genotype and its association with the susceptibility of chronic periodontitis in Uighur patients of Xinjiang.

METHODSGenomic DNA was obtained from buccal swabs of 41 subjects with severe chronic periodontitis (CP), 43 subjects with moderate CP, 49 subjects with mild CP and 92 ethnically matched healthy control individuals. Genotypes of IL-1RN intron 2 VNTR was analyzed by SSP-PCR method. Then compared the differences in distribution of each genotype.

RESULTSA significant over-representation of IL-1RN intron 2 VNTR allele 2 was found in severe chronic periodontitis group.

CONCLUSIONIL-1RN intron 2 VNTR allele 2 may be a risk indicator for the susceptibility of severe chronic periodontitis in Uighur patients of Xinjiang.

Adult ; Aged ; Chronic Disease ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Interleukin 1 Receptor Antagonist Protein ; Male ; Middle Aged ; Minisatellite Repeats ; Periodontitis ; genetics ; Sialoglycoproteins ; genetics

OBJECTIVETo study whether the anti-apoptotic action is reversed by tetrandrine in a combination with vincristine in human breast carcinoma MCF-7 multidrug-resistant cells.

METHODChromatin condensation was observed by co-staining of fluorescent dyes Hoechst 33342 and propidium iodide; and G1 sub-peak was detected by flow cytometry. Apoptotic cells were detected with TUNEL method. Cellular free ca2+ was determined with Fluo-3 staining method.

RESULTTwo types of chromatin condensation were observed after the sensitive and drug-resistant MCF-7 cells were treated with an antitumor drug vincristine 5 mumol.L-1 for 24 h. The number of cell with chromatin condensation was obviously reduced in the drug-resistant cells treated with the same concentration of vincristine, as compared with the sensitive MCF-7 cells. The number of the apoptotic cells was increased by a combination of non-cytotoxic tetrandrine 20 mumol.L-1 and vincristine in both the sensitive and drug-resistant cells, which was confirmed with fluorescent indication and TUNEL method. The increment of introcellular free Ca2+ level in the cells treated with tetrandrine in a combination of vincristine was detected with Fluo-3 staining method.

CONCLUSIONThe anti-apoptotic action of human breast carcinoma MCF-7 cells can be effectively reversed by tetrandrine.

Alkaloids ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Benzylisoquinolines ; Breast Neoplasms ; pathology ; Calcium ; metabolism ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Drug Synergism ; Female ; Humans ; Tumor Cells, Cultured ; Vincristine ; pharmacology