Remodeling and spacing factor-1 (Rsf-1), firstly discovered in 1998 and located in the nucleus, is one of the most commonly amplified gene and its protein molecular weight is about 240 ×103. The interaction between Rsf-1 and human sucrose non-fermenting protein 2 homologue (hSNF2H) changes the cell growth signals and environment, which is inclined to cause the cells loss of the normal regulation, leading to the abnormal proliferation and occurrence of tumors. Recent study has shown that Rsf-1 is closely related to the prognosis of a lot of human malignancies, such as genitourinary system tumors, gastrointestinal tumors, head and neck cancer, non-small cell lung cancer and glioma.

OBJECTIVE: To evaluate the histocompatibility of poly (lactic-co-glycolic acid)/RNA Ⅲ inhibiting peptide (PLGNRIP) sustained release microspheres.METHODS: The crude peptide comprising N to C-terminals was synthesized using Fmoc method. The crude synthetic RNAⅢ peptide was purified by reverse phase high performance liquid chromatography, followed by component harvesting according to ultraviolet absorption peak, and freeze-drying. PLGNRIP sustained release microspheres with a diameter of 50-70 pm were prepared using liquid-phase multiple emulsion method. The histocompatibility of PLGNRIP sustained release microscopes were preliminarily evaluated through the use of acute general toxicity test, MTT cytotoxicity test, intramuscular implantation test, sensitivity test, and pyrogen test.RESULTS: Acute general toxicity test results showed that all included animals survived and presented with no toxicosis reaction and obviously changed body mass. MTT cytotoxicity test results revealed that the average relative growth rate of cells from two eluents was over 85%, with cytotoxicity grade 1, which indicated no cytotoxicity. Intramuscular implantation tests showed that at 4 weeks after implantation of RiP powder or PLGNRIP microscopes, no obviously congested, degenerated, or necrotic tissue was observed. All RIP powder and a part PLGNRIP microscopes were degraded. Fibroblasts accounted for a large proportion in all cells. NO inflammatory cell infiltration, involving neutrophits and multinucleated giant celts, was observed. Sensitivity test rasults displayed that the average primary irritation index was 0.38, 0.33, arid 0.31 in the eluent stock solution, 2% dinitoflruorobenzene, and physiological saline-administerd groups, respectively. Pyrogen test results showed that fervescence of each rabbit in the experiment was under 0.5 ℃ and the sum of fervescence was under 1.3 ℃ .This is in coincidence with evaluation criteria of pyrogen test.CONCLUSION: PLGNRIP sustained release microspheras exhibit good histocompatibility.

Objective To investigate the influence of β-catenin gene deletion on Stat-5α phosphorylation in bcr-abl induced leukemia cells. Methods The established conditonal hematopoitic β-catenin knockout mice were used to isolate bone marrow cells. Exogenous bcr-abl fusion gene was transduced to these bone marrow cells by retroviral infection with intent to transfom them to leukemia cells.Immunofluorescence was performed to detect the phosphorylation status of Stat-5α in both β-catenin deletion cells and control cells. bcr-abl transcription and protein levels were evaluated with real-time PCR and western blotting. Results Phosphorylation of Stat-5α was reduced significantly in β-catenin deletion leukemia cells on comparison with control cells despite that total Stat-5α protein showed no obvious changes. Total tyrosine phosphorylation and bcr-abl protein expression were reduced in bcr-abl induced β-catenin deletion CML cells,on the contrary, both of the reduction were not seen in bcr-abl induced β-catenin deletion ALL cells.Conclusion Loss of β-catenin inhibits both Stat-5α phosphorylationin and bcr-abl expression in bcr-abl induced leukemia cells.

OBJECTIVE: To evaluate blood compatibility of polyaiticglycolic acid/RNA Ⅲ inhibiting peptide (PLGA/RIP) delayed release microspheres.METHODS: ① Preparation of PLGA/RIP microspheres: The solid-phase synthesis (Fmoc) method was used to synthesize RIP crude sample from C end to N end; the synthesized crude peptide was purified by the reverse phase high performance liquid chromatography. According to UV absorption peak, the components were collected and freeze-dried, to obtain RIP purifications. Then liquid-phase multiple emulsion method was used to prepare PLGA/RIP microspheres at the diameter of 50-70 μm. ② Preparation of eluent: The PLGA/RIP microsphere powders were eluted with sterile physiological saline at 37 ℃, to prepare 1 g/L eluent; then 0.5 g/L eluent was obtained adding equal volume of sterile physiological saline. The hemolysis test, blood clotting test, and platelet aggregation test were conducted to measure prothrombin time and activated partial thromboplastin time, to observe the influence on rabbit leucocytes, erythrocytes and thrombocytes, and to preliminarily evaluate the blood compatibility of PLGA/RIP microspheres. RESULTS: ①The haemolysis rates of eluent stock solution and 0.5 g/L eluents were 3.24% and 2.67% respectively, which were in coincidence with the criteria of medical biomaterials, less than 5%. ② The eluent stock solution and 0.5 g/L eluents of PLGA/RIP microspheres had no significant effect on rabbit clotting time, prothrombin time and activated partial thromboplastin time, the number of rabbit leucocytes, erythrocytes and thrombocytes, as well as platelet aggregation.CONCLUSION: PLGA/RIP delayed release microspheres have a good blood compatibility.

BACKGROUND: Staphylococci inhibitor, RNAⅢ inhibiting peptide, has been firstly synthesized in China, while RNAIII inhibiting peptide/bioinspired phosphorylcholine-based cytomembrane coating (RIP/PC) has been also prepared.OBJECTIVE: To evaluate the blood compatibility of RIP/PC.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at Institute of Clinical Pharmacologic Research and Medical Animal Experimental Center of General Hospital of Chinese PLA from October 2005 to October 2007.MATERIALS: A total of 30 healthy adult New Zealand rabbits were provided by Animal Experimental Center of General Hospital of Chinese PLA.METHODS: The 316L stainless steel was dip into a mixture of 5 g/L quaternionic copolymer tetrahydrpfuran and 10% RNAⅢ inhibiting peptide to obtain stable polymer coating, which was extracted at density of 1 cm~2/mL in saline at 37 ℃ for 72 hours to get 100% eluent. The same volume of saline was added to make 50% eluent. The effects of RIP/PC on leukocytes, eryfhrocytes,and blood platelet of rabbits were detected via measuring prothrombin time and activated thrombin time during hemolysis test,hemagglutinatin test, and blood platelet aggregation test.MAIN OUTCOME MEASURES: Hemolytic ratio of erythrocytes, clotting time, prothrombin time, activated thrombin time, amounts of leukocytes, erythrocytes, and blood platelet, as well as blood platelet aggregation.RESULTS: Hemolysis test showed that the hemolytic ratio was 3.08% and 1.88% of 100% and 50% eluent, respectively; both the hemolytic ratios were < 5%, suggesting being coincidence with hemolytic requests of biological materials. The hemagglutinatin test showed that both 100% and 50% eluent did not have any effect on clotting time, prothrombin time, activated thrombin time,amounts of leukocytes, erythrocytes, and blood platelet, as well as blood platelet aggregation.CONCLUSION: The firstly synthesized RIP/PC has a good biocompatibility.

OBJECTIVE:To evaluate the histocompatibility of poly(lactic-co-glycolic acid)/RNA Ⅲ inhibiting peptide(PLGA/RIP)sustained release microspheres.METHODS:The crude peptide comprising N to C-terminals was synthesized using Fmoc method.The crude synthetic RNAIII peptide was purified by reverse phase high performance liquid chromatography,followed by component harvesting according to ultraviolet absorption peak,and freeze-drying.PLGA/RIP sustained release microspheres with a diameter of 50-70?m were prepared using liquid-phase multiple emulsion method.The histocompatibility of PLGA/RIP sustained release microscopes were preliminarily evaluated through the use of acute general toxicity test,MTT cytotoxicity test,intramuscular implantation test,sensitivity test,and pyrogen test.RESULTS:Acute general toxicity test results showed that all included animals survived and presented with no toxicosis reaction and obviously changed body mass.MTT cytotoxicity test results revealed that the average relative growth rate of cells from two eluents was over 85%,with cytotoxicity grade 1,which indicates no cytotoxicity.Intramuscular implantation tests showed that at 4 weeks after implantation of RIP powder or PLGA/RIP microscopes,no obviously congested,degenerated,or necrotic tissue was observed.All RIP powder and a part PLGA/RIP microscopes were degraded.Fibroblasts accounted for a large proportion in all cells.No inflammatory cell infiltration,involving neutrophils and multinucleated giant cells,was observed.Sensitivity test results displayed that the average primary irritation index was 0.38,0.33,and 0.31 in the eluent stock solution,2% dinitoflruorobenzene,and physiological saline-administerd groups,respectively.Pyrogen test results showed that fervescence of each rabbit in the experiment was under 0.5℃ and the sum of fervescence was under 1.3℃.This is in coincidence with evaluation criteria of pyrogen test.CONCLUSION:PLGA/RIP sustained release microspheres exhibit good histocompatibility.

BACKGROUNDDrug-resistance of tumor is the main reason of the failure of che-motherapy. Much attention has been paid to the expression of multidrug resistance-associated protein (MRP) and its mechanism of drug-resistance in non-small cell lung cancer (NSCLC). Results of this research will contribute to reversing drug-resistance and improving curative effect. The aim of this study is to investigate the relationships between the expression of MRP and clinicopathological parameters and prognosis in patients with NSCLC.

METHODSExpression of MRP was detected in 62 cases of paraffin-embedded NSCLC samples by streptavidin-biotin-peroxidase complex immunohistochemistry method, as well as in 30 fresh cases of NSCLC samples and corresponding normal lung tissues by immunohistochemistry and Western blot.

RESULTSMRP expression of NSCLC tissues was significantly higher than that of normal lung tissues. The survival time of patients with negative MRP expression was (69.81±17.41) months, and that of patients with positive MPR expression was (25.38±4.46) months (P=0.0156). This statistically significant relationship between the survival time and prognosis was also showed in squamous cell carcinoma patients (P=0.015), but not in adenocarcinoma. Multivariate COX model analysis suggested that the survival time was significantly related to lymphatic metastasis (P=0.038) and expression of MRP (P=0.035).

CONCLUSIONSMRP expression in NSCLC is significantly higher than that in the normal lung tissues. The mean survival time of patients with negative MRP expression is remarkably longer than that of patients with positive MRP expression. MRP expression may be an independent prognostic factor.

BACKGROUNDTo study the expression of MMP-2, MMP-9, TIMP-1 proteins and mRNA in NSCLC and to analyse their relations with prognosis.

METHODSImmunohistochemistry (IHC) and in situ hybridization (ISH) were used to detect the expression of MMP-2, MMP-9, TIMP-1 proteins and mRNA in pa-raffin-embedded NSCLC specimens.

RESULTSThere were no significant differences among the MMP-2, MMP-9, TIMP-1 expression and age, sex, histological type and differentiation. There was statistical relationship between expression of MMP-2, MMP-9, TIMP-1 and lymph node metastasis. Multivariate Cox model analysis suggested that the survival time was significantly related to lymph node metastasis, expression of MMP-2 and MMP-9. The results of IHC and ISH suggested that the concordant rates of MMP-2, MMP-9, TIMP-1 proteins and mRNA were of statistical significance (P < 0.01,P < 0.005,P < 0.025).

CONCLUSIONSMMP-2 and MMP-9 are independent factors that affect prognosis, TIMP-1 is an useful parameter to the prognosis of NSCLC.

BACKGROUNDTo study the expression of MGMT and its relationship with efficacy of chemotherapy and prognosis in patients with non-small cell lung cancer (NSCLC).

METHODSMGMT was detected in 128 NSCLC tissues and 10 normal pulmonary tissues by immunohistochemistry. According to the level of MGMT , 128 patients with NSCLC were divided into the group Mer- with negative MGMT expression and the group Mer+ with positive MGMT expression.

RESULTSMGMT positively expressed in 61 of 128 patients with NSCLC (47.66%), but none of normal group. No significant relationship was found among MGMT expression and TNM stages, lymph node metastasis and histological classification of the cancer. However, the mean survival periods and survival rates in group Mer- were significantly higher than those of group Mer+ (P < 0.01 , P < 0.05). In 45 evaluable patients, total response rates were 42.86% and 4.17% in Mer- and Mer+ patients respectively (P < 0.001); and there were remarkably longer mean survival periods and higher survival rates in Mer- patients than those in Mer+ patients (P < 0.01, P < 0.05).

CONCLUSIONSExpression of MGMT may be helpful to predict efficacy of chemotherapy and prognosis in patients with NSCLC.

OBJECTIVETo investigate the relationship between vascular endothelial growth factor C (VEGF-C) expression, VEGFR-3 expression, lymphangiogenesis and angiogenesis in human non-small cell lung cancer (NSCLC).

METHODSSeventy-six NSCLC samples were stained for VEGF-C, VEGFR-3 and CD34 with immunohistochemical methods. Assessment of lymphatic vessel density (LVD) and microvessel density (MVD) was performed. The expressions of VEGF-C in 24 fresh NSCLC samples were determined with Western blot assay.

RESULTSOf the 76 NSCLC cases, 55 were VEGF-C positive and 40 were VEGFR-3 positive in cancer cells. A significant positive correlation was found between VEGF-C expression and VEGFR-3 expression in cancer cells (P < 0.05). VEGF-C expression was negatively associated with differentiation of tumor cells (P < 0.05). VEGF-C expression and VEGFR-3 expression were positively associated with lymph node metastasis and lymphatic invasion (P < 0.05). LVD was positively related to VEGF-C expression, lymph node metastasis, lymphatic invasion and clinical stage (P < 0.05). There was a significant correlation between LVD and MVD (R = 0.732, P < 0.05). Patients with positive VEGF-C expression had worse outcomes than those with negative VEGF-C expression (P < 0.01).

CONCLUSIONSIn NSCLC, VEGF-C and VEGFR-3 are related to the lymphangiogenesis, angiogenesis, and occurrence and development of lung cancers. VEGF-C expression could be a useful predictor of poor prognosis in NSCLC.

Carcinoma, Non-Small-Cell Lung ; metabolism ; Endothelial Growth Factors ; biosynthesis ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; Vascular Endothelial Growth Factor C ; Vascular Endothelial Growth Factor Receptor-3 ; biosynthesis