OBJECTIVE: To investigate the effect of oxidative stress on the accumulation of heat shock protein 70 (HSP70) within C2C12 myogenic cells. METHODS: Heat shock response (42 degrees C for 1 h and recovery for 12 h at 37 degrees C) was used to induce the expression of heat shock protein 70. We constructed a recombinant plasmid of HSP70 with enhanced green fluorescent protein (EGFP). After being transfected transiently into C2C12 cells, immunoblotting was used to detect the expression of HSP70 induced by heat shock response and transfection. Immunocytochemistry, fluorescent microscopy and immunoblotting were used to detect the translocation of HSP70. RESULTS: Immunoblotting showed that the overexpression of HSP70 was induced by heat shock response and transient transfenction. HSP70 localized within the cytoplasm of the normal cells, but HSP70 translocated from the cytoplasm to the nucleus and the nucleolus at 1 h after the treatment of oxidative stress (0.5 mmol/L H2O2) by using immunocytochemistry, fluorescent microscopy and immunoblotting for cellular partial proteins. CONCLUSION Oxidative stress may induce the accumulation of heat shock protein 70 within the nucleolus.
Cell Nucleolus ; metabolism ; Cells, Cultured ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Myoblasts ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Oxidative Stress ; physiology

Serum enzymes that play potential roles in tumor growth have recently been reported to have prognostic relevance in a diverse array of tumors. However, prognosis-related serum enzymes are rarely reported for nasopharyngeal carcinoma(NPC). To clarify whether the level of serum enzymes is linked to the prognosis of NPC, we reviewed the pretreatment data of lactate dehydrogenase(LDH), alkaline phosphatase (ALP), and glutamyl transferase (GGT) in 533 newly diagnosed NPC patients who underwent radical radiotherapy between May 2002 and October 2003 at Sun Yat-sen University Cancer Center. Patients were grouped according to the upper limit of normal values of LDH, ALP, and GGT. The Kaplan-Meier method and log-rank test were used for selecting prognostic factors from clinical characteristics and serum enzymes, and the chi-square test was applied to analyze the relationships of clinical characteristics and serum enzymes. Finally, a Cox proportional hazards model was used to identify the independent prognostic factors. We found that increased levels of LDH had poor effects on both overall survival and distant metastasis-free survival (P = 0.009 and 0.035, respectively), and increased pretreatment level of serum ALP had poor effects on both overall survival and local recurrence-free survival (P = 0.037 and 0.039, respectively). In multivariate analysis, increased LDH level was identified as an independent prognostic factor for overall survival. Therefore, we conclude that increased pretreatment serum LDH and ALP levels are poor prognostic factors for NPC.
Adolescent ; Adult ; Aged ; Alkaline Phosphatase ; blood ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Cisplatin ; administration & dosage ; Female ; Fluorouracil ; administration & dosage ; Humans ; Kaplan-Meier Estimate ; L-Lactate Dehydrogenase ; blood ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; blood ; drug therapy ; pathology ; radiotherapy ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Prognosis ; Proportional Hazards Models ; Radiotherapy, Computer-Assisted ; Radiotherapy, Conformal ; Radiotherapy, Intensity-Modulated ; Survival Rate ; Young Adult ; gamma-Glutamyltransferase ; blood

OBJECTIVETo investigate the clinical characteristics, international prognostic index and treatment of primary non-Hodgkin's lymphoma (NHL) of the nasopharynx.

METHODSFrom January 1983 to December 1997, 136 patients with previously untreated NHL of the nasopharynx were retrospectively reviewed. All patients were confirmed pathologically and classified by Working Formulation system. There were 18 patients with high-grade, 77 intermediate, 2 low-grade and 39 unclassifiable lymphoma. According to Ann Arbor classification, 25 patients had stage I, 91 stage II, 12 stage III and 8 stage IV lesions. Primary therapy was radiotherapy alone in 13 patients and radiotherapy combined with chemotherapy in 12 patients with stage I disease. In 88 patients with stage II, radiotherapy alone was given to 31 patients, and a combination of radiotherapy and chemotherapy to 57 patients. Chemotherapy was primary treatment for advanced stage III/IV diseases.

RESULTSThe overall survival rate (OS), cancer specific survival rate (CSS) and disease-free survival rate (DFS) at 5 and 10 years for all patients were 56.2%, 61.2%, 51.1% and 48.3%, 58.0%, 46.5%, respectively. As for international prognostic index (IPI), the 5-year CSS was 70.9% for 0 risk factor, 44.9% for 1 risk factor, 30.0% for 2 or 3 risk factors, respectively (P = 0.004). For stage I patients, the 5-year CSS was 83.1% for RT alone and 82.2% for combined modality therapy, respectively (P = 0.779). For patients with stage II, the 5-year CSS was 46.0% for radiotherapy alone and 70.9% for combined modality therapy. There was significant difference between them (P = 0.04). Multivariate analysis by Cox regression showed that Ann Arbor stage, B symptom and IPI were independent prognostic factors.

CONCLUSIONInternational prognostic index is an important prognostic factor for Non-Hodgkin's lymphoma of the nasopharynx and the combined modality therapy may be optimal for the stage II patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; therapeutic use ; Bleomycin ; administration & dosage ; Child ; Combined Modality Therapy ; Cyclophosphamide ; administration & dosage ; Disease-Free Survival ; Doxorubicin ; administration & dosage ; Female ; Humans ; Lymphoma, Non-Hodgkin ; drug therapy ; radiotherapy ; therapy ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; drug therapy ; radiotherapy ; therapy ; Neoplasm Staging ; Prednisone ; administration & dosage ; Prognosis ; Retrospective Studies ; Survival Rate ; Vincristine ; administration & dosage

This study aimed to explore the potentiating effect and mechanism of the extract of Jingfang Granules(JFG) on the activation of macrophages. The RAW264.7 cells were treated with JFG extract and then stimulated by multiple agents. Subsequently, mRNA was extracted, and reverse transcription-polymerase chain reaction(RT-PCR) was used to measure the mRNA transcription of multiple cytokines in RAW264.7 cells. The levels of cytokines in the cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA). In addition, the intracellular proteins were extracted and the activation of signaling pathways was determined by Western blot. The results showed that JFG extract alone could not promote or slightly promote the mRNA transcription of TNF-α, IL-6, IL-1β, MIP-1α, MCP-1, CCL5, IP-10, and IFN-β, and significantly enhance the mRNA transcription of these cytokines in RAW264.7 cells induced by R848 and CpG in a dose-dependent manner. Furthermore, JFG extract also potentiated the secretion of TNF-α, IL-6, MCP-1, and IFN-β by RAW264.7 cells stimulated with R848 and CpG. As revealed by mechanism analysis, JFG extract enhanced the phosphorylation of p38, ERK1/2, IRF3, STAT1, and STAT3 in RAW264.7 cells induced by CpG. The findings of this study indicate that JFG extract can selectively potentiate the activation of macrophages induced by R848 and CpG, which may be attributed to the promotion of the activation of MAPKs, IRF3, and STAT1/3 signaling pathways.
Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/metabolism* ; Plant Extracts/metabolism* ; Lipopolysaccharides/pharmacology* ; Macrophages ; Cytokines/metabolism* ; RNA, Messenger/metabolism*