ObjectiveTo investigate the activation of MAPK signal pathway in multiple myeloma and the regulation of BLyS expression levels through MAPK signal pathway; preliminarily study the role of MAPK signal pathway in the up-regulation of BLyS expression levels induced by IFN-γ.MethodsActivated MAPK pathway were detected by Western blot,while the expression of BLyS were detected with RT-PCR and Western blot,and Western blot investigated the effect of MAPK pathway on BLyS expression levels induced by IFN-γ.ResultsIn addition to the expression of ERK,JNK,p38,p-JNK was also expressed in MM cell lines,the MAPK pathway inhibitor targeting JNK SP600125 can down-regulate the expression of BLyS,and its activator anisomycin can up-regulate the expression of BLyS.SP600125 restrained the proliferation and survival of MM cells.ConclusionJNK/SAPK pathway was activated in MM cells; The activated degree of JNK/SAPK pathway and the expression level of BLyS was positively correlated.JNK/SAPK pathway play an important role in the up-regulation of BLyS expression levels induced by IFN-γ.

Astrocytoma ; genetics ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Frizzled Receptors ; genetics ; metabolism ; Glioblastoma ; genetics ; metabolism ; Glioma ; genetics ; metabolism ; Humans ; Paraffin Embedding ; RNA, Messenger ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Signal Transduction ; Wnt2 Protein ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

OBJECTIVETo study the effects and mechanism of hydrogen peroxide (H2O2) of low concentration on dynamic changes of intracellular free calcium contents ([Ca2+]i) in cultural rat liver oval cells (WB-F344 cells).

METHODSUsing Fluo-3/Am as fluorescent indicator of [Ca2+]i and it was measured by laser scanning confocal microscope system.

RESULTSThe results showed that: (1) A rapid transient spiking of [Ca2+]i occurred after the stimulation of H2O2 of low concentration (800 nmol/L). (2) The [Ca2+]i increase was abolished by pretreated with catalase (CAT) or by incubated in D-Hank's solution containing EGTA, the chelate of extracellular Ca2+. (3) The [Ca2+]i increase was not inhibited by pretreated nifedipine, Ca2+ channel blocker, but was abolished by pretreated with anthracere-9-cardoxylic acid (A9C), the Cl-channel blocker and which also blocked calcium activated non-selective cation channel (CAN).

CONCLUSIONSThese results suggest that the increase of [Ca2+]i induced by H2O2 of low concentration may be due to the influx of extracellular Ca2+ through CAN.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Hepatocytes ; metabolism ; Hydrogen Peroxide ; pharmacology ; Ion Channels ; drug effects ; Microscopy, Confocal ; Rats

AIMTo study the action of RMP-7 and its derivative on transporting liposome across the blood brain barrier (BBB) into the brain.

METHODSRMP-7 and DSPE-PEG-NHS [[1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-n-[poly (ethylene-glycol)]-hydroxy succinamide]] were conjugated together in mild condition and MALDI-TOF-MS (Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry) was used to determine their molecular ratio. An in vitro BBB model was established and used to determine in vitro bioactivity of RMP-7 and its derivative. The fluorescence of brain slices and the Evens Blue (EB) concentration in the brain, liver, spleen, lung and kidney of each group were used to evaluate the in vivo bioactivity of RMP-7 and its derivative on transporting liposome across the BBB.

RESULTSThe average molecular weight (MW) of the reaction product was 4,900, while those of DSPE-PEG-NHS and RMP-7 were 3,224 and 1,098. The results demonstrated that RMP-7 was conjugated to DSPE-PEG-NHS at the molecular ratio of 1:1, so the product was DSPE-PEG-RMP-7. RMP-7 and DSPE-PEG-RMP-7 was shown to improve the transporting of peralcohol enzyme across the in vitro BBB model 2-3 times higher than the peralcohol enzyme only. DSPE-PEG-RMP-7 could facilitate the transporting of EB into brain more easily than RMP-7.

CONCLUSIONBoth RMP-7 and DSPE-PEG-RMP-7 could facilitate the transporting of liposome across the BBB, especially DSPE-PEG-RMP-7.

Animals ; Biological Transport ; Blood-Brain Barrier ; drug effects ; Bradykinin ; analogs & derivatives ; pharmacology ; Brain ; metabolism ; Drug Carriers ; Drug Delivery Systems ; Evans Blue ; pharmacokinetics ; Liposomes ; pharmacokinetics ; Phosphatidylethanolamines ; Polyethylene Glycols ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution

OBJECTIVETo investigate regulation function of anodonta glucan HBP-A on chondrocytes through Wnt pathway in vitro.

METHODSRat chondrocytes were cultured and differentiated induced with IL-1beta (10 ng/ml) in vitro. Chondrocytes were divided into five groups:IL-13 group,IL-1beta + IWP-2 (5 microM,Wnt pathway inhibitor) group, IL-1beta + HBP-A (0.3 mg/ml) group and IL-1beta + IWP-2 + HBP-A group. Wnt-3a, beta-catenin (24 h,48 h,72 h) and MMP-13(72 h) genes expression were detected by Rt-PCR, while beta-catenin, MMP-13, Sox-9 and coll-II (48 h) protein expression were measured by Western-blot.

RESULTSAfter induction of IL-1beta, gene expression of Wnt-3a, beta-catenin and MMP-13 were increased,so were the protein expression of beta-catenin and MMP-13. In contrast,protein expression of Sox-9 and Coll-II were declined. Following addition of HBP-A, Wnt-3a, beta-catenin and MMP-13 were shown as induction of IL-1beta, but protein expression of Sox-9 and Coll-II were upgraded. Combining HBP-A with IWP-2 led to the lowest level in Wnt-3a, beta-catenin gene and beta-catenin protein expression and highest expression of Sox-9 protein.

CONCLUSIONHBP-A could not only delay the differentiation of chondrocytes through downgrading the signal expression of Wnt/beta-catenin,but also adjust the expression of Wnt-3a, beta-catenin and Sox-9 when combinated with the Wnt inhibitor.

Animals ; Anodonta ; chemistry ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; metabolism ; Glucans ; pharmacology ; Interleukin-1beta ; metabolism ; Rats ; Wnt Signaling Pathway ; drug effects ; Wnt3A Protein ; genetics ; metabolism ; beta Catenin ; metabolism

Objective To investigate a reasonable and effective internal fixation method for posterolateral fracture of the tibial plateau. Methods Specimens of the tibial plateau with posterolateral fracture made from 12 adult male cadavers were randomly and evenly divided into 3 groups, and fixed by anterior 6.5 mm lag screw, lateral 4.5 mm L-shape plate, posterior 3.5 mm T-shape plate, respectively. All the specimens were loaded in turn by stress of 250, 500, 750, 1 000 N, and the corresponding axial displacement and stress were measured. Results Under the same stress, the Y-axial displacement of the anterior lag screw group was the smallest, showing a significant difference with the lateral plate group and the posterior plate group, while there was no significant difference between the lateral plate group and the posterior plate group in the Y-axial displacement. The stresses on marked points in the anterior lag screw group were evenly distributed. Conclusions For fixation of isolated posterolateral fractures of the tibial plateau, the anterior 6.5 mm lag screw can effectively increase the axial stability and balance the stress distribution around the fracture block, indicating it is an effective method for mechanical fixation. The lateral plate has certain advantage in lateral stability control, while the posterior plate has certain value to reduction of the posterior tibia plateau fracture.

OBJECTIVETo study the anti-invasion effect of SEPT7 gene on U251MG glioma cells and its possible molecular mechanism.

METHODSRecombinant adenovirus vector carrying SEPT7 gene (rAd5-SEPT7) was transduced to human glioma cell line U251MG, and empty adenovirus vector was used as control. Tumor invasion was examined by Transwell method and 3 D-Matrigel assay, and tumor cell migration by wound-healing method and 2 D-Matrigel assay. Three major molecular events associated with cell motility and migration, including changes of expression in MMP2, MMP9, MT1-MMP, TIMP1 and TIMP2, the alteration of integrin alpha(v)beta(3) expression, and the structural change of cytoskeleton protein, tubulin-alpha, in U251 cells transduced with rAd5-SEPT7 were studied by Western blotting, immunofluorescence and laser scanning confocal microscope, respectively.

RESULTSThe invasive and migratory capabilities of cells transduced with rAd5-SEPT7 were inhibited. The expression of extracellular matrix metalloproteinases MMP-2, MMP-9, MT1-MMP and integrin alpha(v)beta(3) was significantly decreased, while the expression of matrix metalloproteinase inhibitor TIMP1, TIMP2 was upregulated. Intracellular cytoskeleton protein-tubulin-alpha in U251 cells exhibited prominent morphological changes which including the appearance of distortion and aggregation resulting from redistribution of tubulin-alpha, and this feature of alteration was similar to the tubulin-alpha structure in normal non-tumor cells.

CONCLUSIONSEPT7 gene can inhibit the invasion and migration ability of U251 glioma cells. Its molecular mechanism may include that SEPT7 gene reverses the imbalanced state of MMPs/TIMPs, downregulates the expression of integrin alpha(v)beta(3) and alters the structure of tubulin-alpha of U251MG glioma cells. It is suggested that SEPT7 gene could be a good candidate for gene therapy of gliomas.

Adenoviridae ; genetics ; Blotting, Western ; Cell Cycle Proteins ; genetics ; physiology ; Cell Line, Tumor ; Cell Movement ; genetics ; Genetic Vectors ; genetics ; Glioma ; metabolism ; pathology ; Humans ; Integrin alphaVbeta3 ; metabolism ; Matrix Metalloproteinase 14 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Microscopy, Confocal ; Neoplasm Invasiveness ; genetics ; Septins ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

Objective To study the suppressive effect of knockdown of miR-21 on the U87 human giioma xenograft growth and the possible mechanism. Methods Nude mice bearing U87 human glioblastoma subcutaneously were treated with miRNA-21 anfisense oligonucleotides(AS-miR-21)intratumomlly every 3 d until the observation peded ended.The tumor volume of the mice treated withAS-miR-21 was measured regularly as compared with that in the control untreated mice and in the mice treated with scramble oligonucelotides(ODN).Finally,the tumors were removed from nude mice for the examination.In-sire hybridization and real-time PCR were conducted to detect the miRNA expression of miR-21.The biological charaetedsties of the tumors were evaluated by HE and immunohistochemieal staining, and the cell apoptosis was detected by TUNEL method. Resulls During the observation period,the tumor growth was delayed and the final tumor volume of AS-miR-21 heated group was smaller than that in the control and scramble ODN treatedg roup(F=6-056,P=0.007).The expression of miRNA precursor was knocked down in As-miRNA treated tunlors compared with that in untreated or scramble ODN treated tumors.Histopathological examination exhibited the appearance of degraded malignancy.The expressions of PCNA and MMP-9 were down-regulated while Septin-7 and P21 were up-regulated and apoptotic index was increased significantly (F=141.021,P=000) as well.Conclusion The suppressive effect of anti-miR-21 ODNs on the growth of U87 human glioma xenogratts is significant and miR-21 Call be taken as a candidate for gene therapy ofhuman glioma.

OBJECTIVETo study the diagnosis and treatment of a second primary malignant tumor induced by previous radiotherapy.

METHODSFrom March 1970 to March 1997, 108 nasopharyngeal cancer (NPC) patients who developed a second primary malignant tumor induced by radiotherapy were treated. There were squamous carcinoma 43 (39.8%), sarcoma 26 (24.1%), malignant fibrous histiocytoma 14 (13.0%), adenoid cystic carcinoma 12 (11.1%), thyroid papillary adenocarcinoma 8 (7.4%) and malignant melanoma 5 (4.6%). Fifty patients underwent operation, 32 received radiotherapy, 18 received chemotherapy and 8 received operation combined with chemotherapy.

RESULTSThe 3- and 5-year tumor-free survival rates were 64.0% and 36.0% in the operation group. They were 34.4% and 18.8% in the radiotherapy group.

CONCLUSIONSurgery, if not contra-indicated, is the first choice for the second primary malignant tumor induced by radiotherapy. Aggressive treatment for these patients is, hence, indicated clinically.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasms, Radiation-Induced ; diagnosis ; mortality ; therapy ; Neoplasms, Second Primary ; diagnosis ; mortality ; therapy ; Radiotherapy ; adverse effects ; Survival Rate

OBJECTIVETo analyze the correlation between prognosis and p53 expression in primary lesion and the surgical margin of laryngeal squamous cell carcinoma (SCC) as an indication of postoperative radiotherapy.

METHODSSixty-seven laryngeal SCC with pathological negative margin were analyzed retrospectively. Immunohistochemical method was used to detect the expression of p53.

RESULTSThe p53 positive rates in the primary tumor and the surgical margin were 19.4% (13/67) and 50.7% (34/67). In p53 positive primary tumor group, the survival rate was higher in patients who received postoperative radiotherapy than those without (60.6% vs 20.0%, P = 0.000 5) and the recurrent rate was just the reverse (42.1% vs 93.3%, P = 0.002), though these differences were not significant in p53 negative primary tumor group (87.5% vs 94.1%, P = 0.409 6; 25.0% vs 5.9%, P = 0.175). The recurrent rate and survival rate between patients with and without postoperative radiotherapy did not show any significant difference either in p53 positive surgical margin group (47.4% vs 20.0%, P = 0.378 1; 62.5% vs 80.0%, P = 1.0) or p53 negative ones (84.9% vs 66.6%, P = 0.074 3; 20.6% vs 40.7%, P = 0.248).

CONCLUSIONPostoperative radiotherapy should be given to patients with p53 positive primary laryngeal cancer. But those who are pathologically margin negative but p53 positive should not be taken, at least for the present, as candidates for postoperative radiotherapy.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; chemistry ; mortality ; radiotherapy ; Female ; Humans ; Laryngeal Neoplasms ; chemistry ; mortality ; radiotherapy ; Male ; Middle Aged ; Survival Rate ; Tumor Suppressor Protein p53 ; analysis