AIM: The current study was designed to construct eukaryotic expression vector containing NK4 gene and transfect it into human pancreatic cancer cell lines.METHODS: The recombinant of pcDNA3/hNK4 was digested by restriction enzyme, the NK4 gene was cloned into a high effective eukaryotic expressing plasmid which contains CMV2 immediate early gene promoter and then transiently introduced into the pancreatic cancer cell line SW1990 by lipofectamine and clonal cell lines that secrete high levels of NK4 protein were isolated.The expression of NK4 was observed by RT-PCR and Western blot, in vitro the vascular endothelial cell proliferation inhibiting activity of NK4 was examined by 3-[4,5-dimethylthiazolzyl]-2,5-diphenyl tetrazolium bromide(MTT) method. RESULTS: A specific expression of NK4 gene mRNA by lipofectamine-mediated transfer exhibited only in SW1990/NK4 cells,Western blot analysis demonstrated that there was positive expression of NK4 protein(50 kD).The NK4 inhibited proliferation of the vascular endothelial cells in vitro. CONCLUSION: The recombinant of pRC/CMV2-hNK4 is a high effective expressing eukaryotic vector.The bio-engineering product of the NK4 is an angiogenesis inhibitor and may play an important role in the gene therapy for tumor.

Objective To evaluate the tumor targeting characteristic of the Folate-SPIO-DOX-Micelles by in vitro studies,and to test the feasibility of monitor tumor targeting using it and clinical MRI.Methods The polymeric micelles,Folate-SPIO-DOXO-Micelles were prepared.The in vitro tumor cell targeting efficacy of these folate modified and DOX or SPIO-loaded micelles (Folate-SPIO-DOX-Micelles)was evaluated by observing the cellular uptake of micelles by human hepatic carcinoma cells(Bel 7402 cells) which overexpressed folate surface receptors. Cell suspensions were incubated with Folate-SPIO-DOXO-Micelles for 1 h.Prussian blue staining was performed to show intracellular irons.Flow cytometry was used to further quantify the cellular uptake of the nanoparticles into Bel 7402 cells.MRl was performed to show the signal intensity changes by using T2 WI sequences at a clinical 1.5 T MR system.Results Prussian blue staining showed much more intracellular iron in cells incubated with Folate-SPIO-DOX-Micelles than the cells incubated with the non-targeting SPIO-DOX-Micelles.As revealed by flow cytometry,the mean fluorescence intensity of cells in the folate group and the non-folate group were 117.88 and 46.33,respectively.The T2 signal intensity in MRI of cells treated with the folate targeting micelles decreased significantly (when the concentration of SPIO in cell culture medium was 5,10,20,40,and 80 μg/ml,respectively,T2 signal intensity decreased by -5.02%,-23.58%,-45.89%,-70.34%,and -92.41%,respectively).In contrast,T2 signal intensity did not show obvious decrease for cells treated with the folate-free micelles (when the concentration of SPIO in cell culture medium was at 5,10,20,40,and 80 μg/ml,respectively,T2 signal intensity decreased by -3.77%,-2.16%,-2.18%,-2.74% and -19.77%,respectively).Conclusion The polymeric micelles,Folate-SPIO-DOX-Micelles has good targeting ability to the hepatic carcinoma cells in vitro,and the cell targeting events of the micelles can be monitored by using a clinical MR scanner.

OBJECTIVETo detect the expression of hypoxia-inducible factor-1alpha(HIF-1alpha) in endometriosis and explore the possible role of HIF-1alpha in the pathogenesis of endometriosis.

METHODSImmunohistochemistry was performed to examine the expression of HIF-1alpha in 20 normal endometrium, 20 ectopic endometrium and 68 eutopic endometrium specimens from 68 endometriosis patients, and the results were analyzed statistically.

RESULTSThe expression of HIF-1alpha was significantly increased in ectopic endometrium than in normal endometrium (P<0.01), and the expression did not undergo changes with the normal menstrual cycle in the three types of endometrium.

CONCLUSIONHIF-1alpha expression increases in ectopic endometrium, suggesting that HIF-1alpha plays an important role in the pathogenesis of endometriosis.

Adult ; Endometriosis ; genetics ; Endometrium ; metabolism ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Menstrual Cycle ; metabolism