OBJECTIVETo investigate the expression and significance of peroxisome proliferators-activated receptor gamma (PPAR gamma) in human glioma.

METHODSImmunohistochemical staining for PPAR gamma was performed using biopsy specimens of human glioma of various histological types. Expression of PPAR gamma and GFAP in glioma cell lines SWO-38, U251 and SHG-44 were analyzed using Western blotting and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSImmunohistochemical study showed that PPAR gamma was expressed in glioma tissues with positive rate of 37.5%. Western blotting and RT-PCR showed that PPAR gamma was expressed in both glioma cell lines SWO-38 and U251, but not in SHG-44 cells. However, high expression of GFAP was detected in SHG-44 cells.

CONCLUSIONPPAR gamma is associated with carcinogens of glioma. Actived PPAR gamma by agonist may be a novel approach to the treatment of glioma.

Blotting, Western ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Glial Fibrillary Acidic Protein ; biosynthesis ; genetics ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; PPAR gamma ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate whether the deletion of PTEN affects the expression of Cu/Zn SOD and the related biology.

METHODSProtein and mRNA expression levels of PTEN, P-Akt, Cu/Zn SOD in the control immortalized wild type mouse embryonic fibroblast cells (PTEN+/+) and PTEN-null cells (PTEN-/-) were evaluated by Western blot and Northern blot respectively. The level of superoxide anions were detected using fluorescent probes. The DNA damage was documented by single cell alkalescence gel assay. MTT was used to study the effect of H2O2 on the proliferation of cells.

RESULTSThe expression of Cu/Zn SOD was down regulated at both protein and mRNA levels, and the level of superoxide anions increased in the PTEN-null cells (PTEN-/-). The phosphorylation level of Akt kinase was up-regulated and the antiproliferative effect of H2O2 decreased in PTEN-/- cells. Furthermore, DNA damage was observed significantly severer in both the blank control and H2O2 treated groups than that in the PTEN+/+ cells.

CONCLUSIONSDeletion of PTEN affects the expression of Cu/Zn SOD. As a result, reactive oxygen species (ROS) keep at a high level, along with decrease of accumulated oxidative damage and the antiproliferative effect of ROS.

Animals ; Copper ; chemistry ; Down-Regulation ; Fibroblasts ; pathology ; Gene Expression Regulation ; genetics ; Hydrogen Peroxide ; metabolism ; Mice ; Mice, Knockout ; PTEN Phosphohydrolase ; deficiency ; genetics ; Phosphorylation ; Reactive Oxygen Species ; Sequence Deletion ; Superoxide Dismutase ; genetics ; metabolism ; Zinc ; chemistry

OBJECTIVETo investigate the inhibitory effect of artesunate on human endometrial carcinoma RL95-2 cell line proliferation in vitro and the possible mechanisms.

METHODSThe inhibitory effect of artesunate on the cell proliferation was assessed with MTT assay. Transmission electron miscrosopy was used to observe the morphological change of the cells after the treatment. Flow cytometry was performed to examine the changes in the cell cycle, reactive oxygen species (ROS) levels (with DCFH-DA labeling) and mitochondrial membrane potential (rhodamine123 staining), and caspase-3 activity was detected by immunohistochemistry.

RESULTSArtesunate inhibited the proliferation of RL95-2 cells with an IC(50) of 26.29 microg/ml. Transmission electron microscopy revealed early apoptotic changes of the cells with obvious chromatin fragmentation. The cell cycle arrest at G(0)/G(1) phase was observed by flow cytometry, and immunohistochemistry demonstrated caspase-3 positivity in cytoplasm. ROS generation in the cells increased obviously after treatment with artesunate for 72 h, which also resulted in lowered mitochondrial membrane potential.

CONCLUSIONArtesunate suppressed the proliferation of RL95-2 cells in vitro possibly by inducing cell apoptosis.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Endometrial Neoplasms ; pathology ; Female ; Humans

OBJECTIVETo study the clinicopathologic features, immunohistochemical findings and prognosis of primary diffuse large B-cell lymphoma (DLBCL) of testis.

METHODSFourteen cases of primary DLBCL of testis, diagnosed according to the 2001 World Health Organization staging standards for hematopoietic and lymphoid tumors, were retrospectively studied. Immunohistochemical study was performed and follow-up information analyzed.

RESULTSThe median age of the patients was 62 years. There were 10 patients in stage I, 3 in stage II and 1 in stage IV. Follow-up information was available in 11 patients (78.6%). Three of which were still alive and eight died (with duration of survival ranging from 5 to 19 months). The patients usually presented with unilateral painless enlargement of testis. Histologically, the lymphoma cells of all cases showed a centroblastic appearance. One case belonged to the germinal center B cell-like subtype on immunohistochemical study, while the remaining 13 cases were classified as non-germinal center B cell-like subtype. Ten of the 14 cases (71.4%) showed overexpression of p53 protein. Most cases demonstrated high proliferation index. Six of the 14 cases (42.9%) were positive for bcl-2 protein. The overall 1-year, 2-year and 5-year survival rates were 45.5%, 17.0% and 17.0%, respectively.

CONCLUSIONSMost cases with primary DLBCL of testis were of peripheral activated B-cell origin. The prognosis is usually not favorable, with propensity of local relapse and systemic dissemination. Accurate pathologic diagnosis relies on detailed histologic examination and immunohistochemical study.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Germinoma ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neprilysin ; metabolism ; Orchiectomy ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Retrospective Studies ; Seminoma ; pathology ; Survival Rate ; Testicular Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Tumor Suppressor Protein p53 ; metabolism ; Vincristine ; therapeutic use

OBJECTIVE: To observe the length heteroplasmy and point heteroplasmy in human mtDNA control region. METHODS: The peripheral blood, buccal cell, and single hair shaft from 50 individuals and 16 family members, related in their maternallineage were analyzed by direct sequencing, and clones from 20 individuals whose mtDNA sequences have a T-C transition at 16189 nt were sequenced. RESULTS: No point heteroplasmy were observed in peripheral blood, buccal cell, single hair shaft from the same individual, neither in maternally related individuals. Length heteroplasmy was observed in those individuals with a homopolymeric tract and the different clones from the same individual has different proportions of length variants, but the hair shafts from the same individual were very similar to the measurements made from blood DNA. No length heteroplasmy was observed between different tissues from the same individual. CONCLUSION mtDNA sequences have a characteristic of high consistency and genetic stability, mtDNA sequencing is a suitable tool for forensic applications such as individual identification.
Base Sequence ; DNA Mutational Analysis/methods* ; DNA, Mitochondrial/genetics* ; Epithelial Cells ; Genetic Heterogeneity ; Hair/chemistry* ; Humans ; Mouth/cytology* ; Point Mutation ; Polymorphism, Genetic/genetics*

BACKGROUND: Changes of elastic fibers in middle cerebral artery(MCA)is close related withthe aged cerebrovascular disease.OBJECTIVE: To observe the changes of elastic fibers of MCA in different aging rats.DESIGN: A descriptive and controlled study based on experimental animals.SETTING: Department of anatomy and central laboratory in a university.MATERIALS: Totally 36 healthy Wistar rats with either gender, weighing 200 - 280 g, were selected from the Animal Institute of the third medical military university of Chongqing[certification SCXX (army) 2002-007].INTERVENTIONS: Changes of elastic fibers of MCA of different aging rats were observed with light microscope, transmission electron microscope and image analysis system. MAIN OUTCOME MEASURES: ①) Major outcome: changes of elastic lamella in MCA of different aging rats; ②) Secondary outcome: ultramicrostructural changes of internal lamella under the transmission electron microscope. RESULTS: With the increase of age, the folded extent and quantity of internal elastic lamella were decreased, and the content of elastic fibers were also decreased significantly( P ＜ 0.01 ). However, the ratio of collagen fibers to elastic fibers was increased significantly( P ＜ 0.01 ) . In the aging group above 24 months, the internal elastic lamina thinned, delaminated and disrupted, and the lipid deposited in it. Endothelial cells and smooth muscle cells passed through the internal elastic lamina. CONCLUSION: Changes of elastic fibers may be related with the increased susceptibility to the cerebrovascular disease in aged people.

OBJECTIVEIt was reported previously that genistein could inhibit proliferation of human ovarian carcinoma cell line SKOV(3), but mechanism was not clear. There is a close relationship between EGFR mediated signal transduction pathway and the development of ovarian tumor. This study aimed to investigate the effects of genistein on the EGFR mediated signal transduction pathway and to clarify its mechanisms of proliferation inhibition on human ovarian carcinoma cell line SKOV(3) and its xenograft in nude mice.

METHODSThe expression of c-erbB-2 protein was determined using immunocytochemistry. The expressions of c-jun and c-fos protein were determined using Western blotting. The expression of c-erbB-2, c-raf-1, c-jun and c-fos mRNA were tested by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe expression of c-erbB-2, c-raf-1 and its downstream gene c-jun and c-fos were decreased at mRNA level in the 20 micromol/L genistein group. The expression of c-erbB-2 protein were decreased, its average absorbency (A) were decreased after treatment of SKOV(3) with 20 micromol/L genistein for 48 h, reached at 0.42 +/- 0.02 (P < 0.05). Western blotting demonstrated that the expressions of c-jun and c-fos protein were decreased gradually after being treated with 20 micromol/L genistein for 12 - 72 h.

CONCLUSIONSGenistein could down-regulate the expression of two key genes, c-erbB-2 and c-raf-1 at mRNA and protein level in the EGFR mediated signal transduction pathway, and down-regulate the expression of its downstream nuclear transcription factors c-jun and c-fos at mRNA and protein level. It is suggested that interfering the expressions of some key signal molecules in EGFR mediated signal transduction system by genistein may account for its molecular foundation of proliferation inhibition in ovarian carcinoma.

Animals ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Genes, fos ; Genes, jun ; Genistein ; pharmacology ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Ovarian Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-jun ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-raf ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, Epidermal Growth Factor ; metabolism ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Signal Transduction ; drug effects

Objective To investigate how genistein to inhibit the invasion of human ovarian serous papillary cystadenocarcinoma cell line SKOV3.Methods Millicell chamber and coculture method were used to establish chemotactic migration model in vitro to observe the effect of genistein on directional chemotactic migration movement of SKOV3 cells.The protein expressions of cell surface adhesion molecule CD44v6,matrix metalloproteinases and tissue inhibitor of metalloproteinases MMP-9/TIMP-1 and MMP-2/TIMP-2 were determined by using immunocytochemical method and their mRNA levels were examined by in situ hybridization.Results Compared with control group,the directional chemotactic migration of SKOV3 were significantly decreased after treated with 20 ?mol/L and 40 ?mol/L genistein,reached to(46.9?5.8)% and(28.3?4.7)% respectively(P

Objective To investigate the anti-tumor pathway and the effect of recombinant E.coli. LLO/OVA in C57BL/6 mice. Methods Magnetic sorting of splenic CD11c, CD4+ and CD8+ T cell from immunized mice with E.coli. LLO/OVA and E.coli. OVA vaccination were performed. In addition, the lymphocytes mixed reaction and IL-2 and IFN-? in the supernatant were detected. The percentage of OVA257-264 SIINFEKL specific CD8+ T cell were checked by flow cytometry. B16-OVA melanoma protection and inhibition response in E.coli. vaccinated mice were compared. Results Compared to E.coli. OVA, E.coli. LLO/OVA-vaccinated splenic CD11c cells stimulated proliferation of autogenic CD4+ T cells and IL-2 production of these cells. CD11c cells induced autogenic CD8+T cells proliferation and IFN-? secretion. A great number of OVA257-264 SIINFEKL-specific splenic CD8+ T cells were induced and the number of lung tumor nodules was significantly reduced in E.coli. LLO/OVA vaccinated mice. Conclusion Recombinant LLO, as an effective adjuvant of E.coli. OVA, can induce murine splenic CD11c cells activation and play potential roles on CD4+ and CD8+ T cells proliferation and IL-2 and IFN-? secretion of these cells, induce more OVA-specific CD8+ T cells and stimulate stronger tumor inhibition in vivo of vaccinated C57BL/6 mice.

Objective To investigate the effects of artesunate on human cervical cancer cell line HeLa.Methods The inhibitory effect on cell proliferation was assessed by MTT assay.Transmission electron microscopy and fluorescent staining were applied to demonstrate the presence of apoptosis.Cell cycle,reactive oxygen species(ROS) levels and mitochondrial membrane potential(??m) were examined by flow cytometry,respectively.Expression of caspase-3 was detected by immunohistochemical methods.Results Growth inhibition of ART on HeLa cells was time-and dose-dependent.Apoptotic feature was observed by transmission electron microscopy and fluorescent staining.The blocking of the cell cycle was in S-phase.The expression of caspase-3 in cytoplasm was positive.The generation of ROS increased obviously(P