We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N.brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.

OBJECTIVETo investigate the expression of beta-dystroglycan (beta-DG) and the roles of tissue inhibitor of metalloproteinases (TIMPs) on beta-DG in salivary adenoid cystic carcinoma (SACC).

METHODSbeta-DG in highly lung metastatic cell line ACC-M and lowly lung metastatic one ACC-2 was tested by immunocytochemistry with different concentrations (10, 15, 20, 25 micromol x L(-1)) of TIMPs, and that without the regulation of TIMPs was served as controls. beta-DG was detected in seven specimens of SACC and ten cases of normal salivary gland tissues which were considered as a comparison group by immunohistochemistry.

RESULTSThere was no positive beta-DG immune-staining at the ACC-2 and ACC-M cell lines without TIMPs in the cell culture. beta-DG expressed after the regulation of TIMPs. beta-DG expression was localized predominantly in basement membrane of the acinus, while the negative results were distributed in the carcinoma cells and around the cancer cell nests.

CONCLUSIONBeta-DG is widely expressed by transmembrane protein that plays important roles in connecting the extracellular matrix to the cytoskeleton, the fracture of this structure means that it is easy to invade and transfer, so restoration of beta-DG expression by TIMPs is considered to be critical for successful treatment of SACC.

Carcinoma, Adenoid Cystic ; Cell Line, Tumor ; Dystroglycans ; Humans ; Immunohistochemistry ; Salivary Gland Neoplasms ; Tissue Inhibitor of Metalloproteinases

Background and purpose:Although there is still no standard ifrst line chemotherapy regimen for metastatic gastric cancer (MGC), the doublet and triplet regimens containing platinum and lfuorouracil were most popular worldwidely. The ECF regimen is the classical and one of the most popular treatment choices in this setting, while the marrow suppression, the renal toxicity and poor compliance inhibits its usage. In order to improve its efifcacy and tolerability, this study conducted 2 phaseⅡ trials by modified ECF regimen, the EOF5 regimen (substituting cisplatin with oxaliplatin, shortening the continuous infusion period to 120 h), to treat patients with MGC since 2004. This paper reported the comprehensive results of the 2 studies.Methods:All the patients who enrolled in our previous2 phaseⅡ trials and received EOF5 as ifrst line treatment entered this study. Each patient received the treatment of epirubicin 50 mg/m2 iv d1, oxaliplatin 130 mg/m2 iv gtt d1 and 5-FU 375-425 mg/m2·d-1 civ 120 h, and repeated every 3 weeks. Efifcacy was analyzed every 6 weeks.Results:A total number of 178 patients (all were metastatic patients but 2 advanced patients with unresectable lesions) were included into this study. One hundred and seventy patients were evaluable, and 7 patients (3.9%) achieved complete response (CR), 76 patients (42.7%) achieved partial response (PR), 46.6% patients achieved overall response rate (ORR, CR+PR), and the cases of stable disease (SD) and progressive disease (PD) were 69 (38.8%) and 18 (10.1%), respectively. The median progress free survival (PFS) and overall survival (OS) were 6.0 months (95%CI: 5.2-6.8) and 12.6 months (95%CI: 8.9-16.3), 1-year and 2-year survival rate were 50.9% and 28.0%, respectively. Grade 3, 4 toxicity including: leucopenia (23.0), neutropenia (38.8%), anemia (6.5%), thrombocytopenia (23.5%), nausea/vomiting (14.1%), and peripheral neuropathy toxicity (1.2%). Among 75 patients who received second line treatment, the median survival from second line treatment was 8.0 months (95%CI: 4.8-11.2).Conclusion:EOF5 regimen is a highly effective regimen with moderate and manageable toxicity, and it providesa suitable alternative for the ifrst-line treatment of MGC.

INTRODUCTIONThe current metastatic category (M) of nasopharyngeal carcinoma (NPC) is a "catch-all" classification, covering a heterogeneous group of tumors ranging from potentially curable to incurable. The aim of this study was to design an M categorization system that could be applied in planning the treatment of NPC with synchronous metastasis.

METHODSA total of 505 NPC patients diagnosed with synchronous metastasis at Sun Yat-sen University Cancer Center between 2000 and 2009 were involved. The associations of clinical variables, metastatic features, and a proposed M categorization system with overall survival (OS) were determined by using Cox regression model.

RESULTSMultivariate analysis showed that Union for International Cancer Control (UICC) N category (N1-3/N0), number of metastatic lesions (multiple/single), liver involvement (yes/no), radiotherapy to primary tumor (yes/no), and cycles of chemotherapy (>4/≤4) were independent prognostic factors for OS. We defined the following subcategories based on liver involvement and the number of metastatic lesions: M1a, single lesion confined to an isolated organ or location except the liver; M1b, single lesion in the liver and/or multiple lesions in any organs or locations except the liver; and M1c, multiple lesions in the liver. Of the 505 cases, 74 (14.7%) were classified as M1a, 296 (58.6%) as M1b, 134 (26.5%) as M1c, and 1 was not specified. The three M1 subcategories showed significant difference in OS [M1b vs. M1a, hazard ratio (HR) = 1.69, 95% confidence interval (CI) = 1.16-2.48, P = 0.007; M1c vs. M1a, HR = 2.64, 95% CI = 1.75-3.98, P < 0.001].

CONCLUSIONSWe developed an M categorization system based on the independent factors related to the prognosis of patients with metastatic NPC. This system may be helpful to further optimize individualized care for NPC patients.

Carcinoma ; Humans ; Multivariate Analysis ; Nasopharyngeal Neoplasms ; Neoplasm Staging ; Prognosis