Objective To construct a cDNA library from human nasopharyngeal carcinoma cell HNE 2.Methods The total RNA was separated from human NPC(nasopharyngeal carcinoma)cell HNE 2 and the mRNA was isolated from the total RNA by MagneSphere technique,then the first-strand cDNA was synthesized with oligo(dT) primer containing sfiI site while the double-strand cDNA was amplified through LD-PCR(Long-distance PCR) by SMART technique.The double-strand cDNA was digested by sfiI(IA &IB)restriction enzyme before cDNA size fractionation ,the double-strand cDNA fractionated was ligated into the ?TripIEx2 vector and then was packaged in vitro.Results The unamplified human NPC cell HNE 2 cDNA library consists of 0 78?10 6 independent clones,and the percentage of recombinant clones was more than 96%.The titer of the amplified cDNA library was 1 02?10 9 pfu/ml and the average insert of the recombinants was 1 2kb.Conclusions The quality of the constructed human NPC cell HNE 2 cDNA library is excellent and helpful to screen NPC specific-antigen.

Objective Through analyzing benign and malignant pleural effusion samples by proteomic techniques, finding protein biomarkers to provide help and new clues for effusion differential diagnosis.Methods Two-dimensional electrophoresis(2-DE) and matrix-assisted laser desorption ionization-time of flight-mass spectrometry(MALDI-TOF-MS) were used to search and identify protein biomarkers, enzyme linked immunosorbent assay(ELISA) was to validate the biomarkers.Results By comparing malignant group with benign group, 43 significantly different protein spots(Up or down regulated≥2 times) were found.Including 9 up regulated spots and 34 down regulated spots.And 7 spots were identified(Up or down regulated≥3 times) by MALDI-TOF-MS and validated 2 spots immunoglobulin λ(Igλ) and haptoglobin(Hp) by ELISA.The results showed that Igλ showed no statistical significance between two groups, while Hp showed the statistical significance(P<0.05).The diagnostic sensitivity and specificity of Hp in malignant pleural effusion were 75.00% and 52.38% at diagnostic cut-off point of 389.02 μg/L.Conclusion The application of proteomics technology has a great help with protein biomarkers searching in pleural effusion.HP has a certain value for the differential diagnosis of benign and malignant pleural effusionand and worthy of further study.

Objective To investigate the effect and potential mechanism of pigment epithelium derived factor(PEDF) acting upon SK-MES-1 cell and human umbilical vein endothelial cells(HUVECs).Methods CCK-8 was used to detect the effect of varying concentrations of PEDF upon HUVECs and SK-MES-1 cell, measuring the degree of cell proliferation and inhibition effect across varying times.The flow cytometry tests were carried out to invest gate the apoptosis of these two kinds of cells when exposed to varying concentration of PEDF.qRT-PCR were carried out to assess the vascular endothelial growth factor(VEGF) gene expression level in these two kinds of cells after treatment of PEDF.Results CCK-8 results revealed that PEDF had a concentration-dependent and time-dependent cell proliferation inhibition effect on SK-MES-1 cell and HUVECs(P<0.05);Flow cytometry showed that the apoptosis of the cells in the treatment group were higher than that of control group(P<0.05), and the apoptosis rate of high concentration group was higher than that of the low concentration group(P<0.05);qRT-PCR results showed that PEDF was able to inhibit expression of mRNA of VEGF in both HUVECs and SK-MES-1 cell compared with control samples(P<0.05).Conclusion The antitumor properties of PEDF is mainly related to the inhibition of tumor angiogenesis and direct effects on tumor cells, the effect of PEDF on HUVECs and SK-MES-1 cell maybe related to the effects of PEDF on downregulating expression of VEGF.

Objective: To investigate the effect of up-regulation of miR-181a expression mediated by constructing miR-181a recombinant eukaryotic expression vector pcDNA3.1(+)-miR-181a on cell proliferation, migration and invasion abilities of esophageal carcinoma cell line TE11. Methods: PCR primers were designed and miR-181a precursor sequence was amplified from 95C cell genomic DNA. The product fragments were cloned into pcDNA3.1(+) to construct the recombinant vector pcDNA3.1(+)-miR-181a. The recombinant eukaryotic expression vector pcDNA3.1(+)-miR-181a was transfected into TE11 cells. The expression of miR-181a mRNA was detected by real-time fluorogenic quantitative-PCR (RFQ-PCR). The effects of pcDNA3.1(+)-miR-181a on cell proliferation, migration and invasion abilities of TE11 cells were detected by MTT, wound healing and Boyden chamber methods, respectively. Results: The miR-181a recombinant eukaryotic expression vector pcDNA3.1(+)-miR-181a was successfully established. RFQ-PCR revealed that the mature miR-181a was able to effectively express in TE11 cells transfected with recombinant vector pcDNA3.1(+)-miR-181a (P<0.05). The overexpression of miR-181a could significantly increase the proliferation, migration and invasion abilities of TE11 cells. Conclusion: Overexpression of miR-181a can increase cell proliferation, migration and invasion abilities of esophageal carcinoma TE11 cells. These results may provide experiment references for further research of the role of miR-181a in cancer development and progression. Copyright© 2011 by Tumor.

Objective To study peripheral blood lymphocytic ?-adrenoceptor(?-AR) densities and ?_1-adrenoceptor gene(?_1-AR mRNA) expression levels in patients with heart failure,to investigate the influences of different ?-adrenoceptor antagonists in ?-AR densities and ?_1-AR mRNA expression levels.Methods 104 cases of patients with heart failure were randomly divided into non-?-adrenoceptor antagonist group(35 cases),metoprolol group(34 cases) and carvedilol group(35 cases),we repeatly determined the ?-AR densities and ?_1-AR mRNA expression levels after two-month therapy.Results The ?-AR densities and ?_1-AR mRNA expression levels in heart failure group reduced significantly compared to these in normal persons(P0.05).After two-month therapy,?-AR densities and ?_1-AR mRNA expression levels in metoprolol group were significantly higher than those in carvendilol group and non-?-adrenoceptor antagonist group(P0.05).Conclusions The peripheral blood lymphocytic ?-AR densities and ?_1-AR mRNA expression level down-regulates in heart failure,which correlate with the severity of heart failure but etiological factors.Metoprolol can up-regulate the ?-AR densities and ?_1-AR mRNA expression level,whereas carvedilol has no such effects.

Objective To study the chemical constituents from the bark of Juglans mandshurica. Methods The compounds were isolated and purified by column chromatography on silica gel,HPLC,and recrystallization.Their structures were elucidated by the physicochemical and spectroscopic evidences. Results Fifteen compounds were identified as:4,8-dihydroxynaphthalenyl-O-?D-(6′-acetoxyl)gluco- pyranoside(Ⅰ),dihydrokaempferol(Ⅱ),juglone(Ⅲ),daucosterol(Ⅳ),kaempferol(Ⅴ),4,8-dihy- droxynaphthalenyl-1-O-?-D-[6′-O-(3″,5″-dimethoxy-4″-hydroxybenzoyl)] glucopyranoside(Ⅵ), kaempferol-3-O-?-L-rhamnoside(Ⅶ),3,3′-dimethoxylellagic acid(Ⅷ),naringenin(Ⅸ),quercetin (Ⅹ),reginolone(Ⅺ),quercetin-3-O-?-L-rhamnoside(Ⅻ),naringenin-7-O-?-D-glucoside(ⅩⅢ),4,8- dihydroxynaphthalenyl-1-O-?-glucoside(ⅩⅣ),4,5,8-trihydroxy-?-tetralone-5-O-?-D-[6′-O-(4″-hy- droxy-3″,5″-dimethoxy-benzoyl)] glucoside(ⅩⅤ).Conclusion CompoundⅠ(juglamanol)is a new compound.CompoundsⅡ,Ⅶ—Ⅸ,Ⅻ,andⅩⅢare isolated from plants of Carya Nutt.for the first time.

Animals ; Extremities ; blood supply ; Ischemia ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; Nitric Oxide ; blood ; Oxidative Stress ; Rats ; Rats, Wistar

Hair Cells, Auditory, Inner ; cytology ; Synapses ; physiology ; ultrastructure

Objective To explore impact of high pulmonary blood flow on the content and metabolism of collagen in rats.Methods Thirty-two male SD rats were randomly divided into shunt group and control group.Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary blood flow.In control group,rats experienced the same expe-rimental processes except the shunting procedure.After 4 and 11 weeks of experiment,these changes of pulmonaryartery collagen Ⅰ,collagen Ⅲ,matrix metalloproteinase(MMP-13)and tissue inhibitor of metalloproteinase(TIMP-1) protein expression of rat were investigated by immunohistochemistry.Results After 4 weeks and 11 weeks of shunt,the collagen Ⅰ,collagen Ⅲ,MMP-13 and TIMP-1 of pulmonary artery in rats of shunt group increased significantly compared with those of control group,respectively(all P

Objective To investigate the expression of vascular endothelial growth factor/vcascular endothelial growth factor receptor(VEGF/VEGFR) in bone marrow of children with acute leukemia(AL),and explore their relationship of the clinical features,and observe changes before and after chemotheropy.Methods The bone marrows of 53 children with AL were assayed to study the expression of VEGF/VEGFR(KDR,Flt-1)before and after chemtheropy with S-P immhistochemical staining.Their relations to the clinical features were evaluated.Results The expressions of VEGF,Flt-1,KDR were significantly higher in newly diagnosed children with AL than those of control group.They were significantly higher in children with acute myeloid leukemia(AML) than in children with ALL.The expressions of VEGF,Flt-1,KDR in remission chidren after chemotheropy were significantly lower than before chemotheropy.There was a positive correlation of the percentage of bone marrow blasts with VEGF expression in children with AL.There was also a positive correlation of the percentage of leukemic cell in blood rountine with VEGF expression.For the untreated group of children,no correlation was found between expressions of VEGF,Flt-1,KDR and age,sex,extramedullary infiltration.Conclusions Expression levels of VEGF,Flt-1,KDR in bone marrow of children with AL increases.VEGF/VEGFR may play an important role in process of chidhood AL.