Objective:To study the effect of ?-3 PUFA on the proliferation and apoptosis of tumor cells transfected with transmembrane TNF? (tmTNF?) expression vector containing peroxisome proliferator responsive element (PPRE). Methods:The tmTNF? expression vector was transfected into HL-60 and MCF-7 tumor cells. The effects of ?-3PUFA and/or exogenous tmTNF? on cell proliferation, cell cycle distribution and apoptosis were measured by growth curve,flow cytometry and DNA ladder assay. Results: The production of exogenous tmTNF?gene was located in the cell membrane. After treatment with?-3 PUFA, the expression of exogenous tmTNF? in transfectant was significantly increased. The proliferation was decreased in HL-60 and MCF-7 cells treated with 6.0?10-5 mol/L ?-3 PUFA,but cell cycle was arrested at G0/G1 and the DNA ladder was detectable only in HL-60 cells. These changes were more obvious in transfected cells than non-transfected cells. Conclusion:?-3 PUFA and the product of exogenous tmTNF? gene induced by ?-3 PUFA could inhibit tumor cell growth more effectually,and induction of apoptosis may play a key role in this process.

Objective To study the apoptosis and its molecular mechanism of MCF-7 cells induced by ?-3 polyunsaturated fatty acid(?-3 PUFA)combined with the product of exogenous transmembrane TNF-?(tmTNF-?) gene.Methods The tmTNF-? eukaryotic expression vector containing the PPRE-tk promoter was transfected into the human breast cancer MCF-7 cells.The effects of ?-3 PUFA and/or exogenous tmTNF-? on the cell proliferation and apoptosis were measured by MTT and DNA ladder assay.The activity of caspase-8 was examined by using special fluorescence substrate,and the expression of caspases(1,8 and 9) were analyzed by RT-PCR and Western blotting.The inhibitor of caspases was used to confirm the function of caspase.Results In 6.0?10~(-5)mol/L ?-3 PUFA-treated MCF-7 cells,only the growth suppression was found.In transfected MCF-7 cells after treated with ?-3 PUFA,not only the proliferation capacity was decreased but the DNA ladder was detectable.The expression changes of caspases(1,8 and 9) and caspase-8 activity were obvious in MCF-7 transfected cells treated with ?-3 PUFA.Growth inhibition and apoptosis induced by ?-3 PUFA and tmTNF-? were partly prevented by the special caspase inhibitor.Conclusion These results suggested that up-regulated expression and activity of caspase might promote MCF7 cells apoptosis induced by ?-3 PUFA and exogenous tmTNF-?,indicating that ?-3 PUFA and exogenous tmTNF-? could cooperate in inhibition of the MCF-7 cell growth and induction of apoptosis.caspase network pathway may play a key role in these processes.

Child ; Humans ; Intestinal Polyps ; genetics ; Molecular Biology

The formation of malignant ascites is concerned with non-obstructive factors including immunity regulator(IL-2,TNF,IFN-?)and factors of inducing blood vessel penetrability(VEGF,MMP) besides obstructive factors. Intraperitoneal chemotherapy makes drug directly act on peritoneum and arrives at the aim of controlling and treating malignant ascites.

Objective To construct the transmembrane TNF-? eukaryotic expression vector in n-3 PUFA dependant manner. Methods PPRE-tk sequence was designed and artificially synthesized, and then inserted it into pcDNA3-TNF( ? 1-12)WB plasmid to construct eukaryotic expression vector pPPRE-tk-tmTNF-? by gene recombine techniques. The tmTNF-? protein expression level was observed in MCF-7 transfected cells incubated with EPA by immunofluorescence technique. Results pPPRE-tk-tmTNF-? expression vector was constructed successfully and identified by agarose gel electrophoretic analysis and nucleotide sequence analysis. EPA could increase tmTNF expression levels in time- and dose-dependant manners. Conclusion tmTNF-? expression vector regulated by n-3 PUFA is successfully constructed.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; blood ; diagnosis ; therapy ; Phosphopyruvate Hydratase ; blood ; Prognosis

Objective To explore the correlation between hyperuricaemia and blood pressure, and blood lipid and glucose. Methods By using simple cluster sampling, 2 branch units from PetroChina Changqing Oilfield Company were selected, and all the 720 subjects with hyperuricaemia (HUA) were assigned to the HUA group; another 620 participants with normal uric acid (UA) level into the normal group. The correlation between blood uric acid and blood pressure,and blood lipid and glucose was assessed by Logistic regression. Results The odds ratio (OR) of those who had 1,2 or 3 abnormal status of hypertension,hyperlipidemia and impaired fasting glucose in the HUA group were much higher than the normal group (OR values were 4. 036,2. 562, and 4. 174, respectively). Logistic regression showed that male, systolic blood pressure ( SBP), GLU, total cholesterol ( TC), triglyceride ( TG), low-density lipoprotein cholesterol (LDL-C) were risk factors of H UA ( OR values were 7. 736,2. 309,1.721,2. 761 and 1. 411,respectively) ,while high-density lipoprotein cholesterol (HDL-C, OR = 0. 211 ) was a protective factor of HUA. Conclusions Gender,blood pressure and blood lipid may have correlation with blood UA. Multiple risk factors should be considered to improve the effectiveness of health education and health promotion.

Objective:To establish the detection method of double-antibody sandwich ELISA about CYFRA 21-1 in human ser-um.Methods:The paired antibody were screened among four strains mAbs of CYFRA 21-1, which was marked by sodium periodate method.The detecting method of double antibody sandwich ELISA was optimizted , and evaluated by specificity , stability and sensitivi-ty.Results:The results showed that a paired of antibody , which was 2F9 as the coated antibody and 6F11 as the labeled antibody, was selected from four mAbs .It was the optimum condition of double antibody sandwich ELISA that the coating antigen concentration of 2F9 was 0.50 μg/ml, while the labeled antibody of 6F11 was diluted 6 000 times.The linear range of standard curve was 0.7-25 ng/ml with r2 =0.990 8, while the limit of detection was 0.666 8 ng/ml, the recovery rate was 98.14%.The cross-reactions with the oth-er analogues in serum were less than 0.1%.The coefficient of variation in group (n=10) was 6.8%, whereas coefficient of variation among group(n=5) was 11.4%.The correlation compared with other foreign ELISA kit was 91.42%.Conclusion:In brief, we suc-cessfully established the method of double antibody sandwich ELISA detecting CYFRA 21-1 level in human serum , laying the foundation for the production of CYFRA21-1 ELISA kit.

Akt1 is a serine-threonine protein kinase that has been implicated in the control of cellular metabolism,survival and growth.Elevated expression of Akt1 has been noted in a significant percentage of human tumors,promoting cellular metastasis.Conversely,some studies have revealed hyperactivated Akt1 inhibited the invasiveness and metastasis of breast cancer cells.To clarify the definite effect of Akt1 on tumorigenesis and development,Akt1 was silenced by RNAi in the highly metastatic murine breast cancer 4T1 cells.Akt1 silencing didn't affect the proliferation of breast cancer cells in MTT assay,while reduced the migration in Transwell assay.Consistent with the above results,Akt1 silencing didn't change the primary tumor weight,but significantly suppressed lung metastasis of 4T1 cells.These observations indicated Akt1 plays an important role in murine breast cancer metastasis,and suggested that Akt1 might be a therapeutic target for breast cancer metastasis.

PURPOSE: To evaluate the morphologic differential diagnosis of benign and malignant ductal breast tumors, as seen on US. MATERIALS AND METHODS: US findings in 29 pathologically proven cases of ductal breast tumor were retrospectively reviewed. All patients were female and their mean age was 42 years. Nineteen tumors were benign and ten were malignant, and all ductal or cystic lesions showed solid masses. According to the location of the mural nodule, we classified the sonographic appearance of these tumors into three types : intraductal, intracystic and amorphic. The intraductal type was divided into three subtypes: incompletely obstructive, completely obstructive and multiple mural nodules. For the intracystic type, too, three subtypes were designated : the intracystic mural nodule (mural cyst), intracystic mural nodule with the duct (mural cyst+duct) and intracystic multiple mural nodules. The amorphic type is defined as an atypical ductal tumor with the mural nodule extending into adjacent parenchyma. RESULTS: The margin of the duct or cyst was smooth in 68.4% of benign, and irregular in 90% of malignant ductal tumors. Internal echogeneity of the duct or cyst usually showed homogeneity in both benign and malignant tumors. 73.7% of tumors connecting the duct were benign and 50% were malignant. In benign tumors, 52.6% of mural nodule had an irregular margin, while in malignant tumors, the corresponding proportion was 100% ; both types usually showed heterogeneous hypoechogeneity. Among benign tumors, the most common morphologic type was the intraductal incompletely obstructive subtype (36.8%) ; among those that were malignant, the amorphic type was most common, accounting for 40% of tumors. No amorphic type was benign and no incompletely obstructive subtype was malignant. CONCLUSION: When ductal breast tumors are morphologically classified on the basis of sonographic findings, the intraductal incompletely obstructive subtype suggests benignancy, and the amorphic type, malignancy. The morphologic classification of ductal breast tumors based on sonography is therefore useful for the differential diagnosis of benignancy and malignancy.
Breast Neoplasms* ; Breast* ; Classification* ; Diagnosis, Differential* ; Female ; Humans ; Retrospective Studies ; Ultrasonography*