Air Pollution ; prevention & control ; Epidemiologic Research Design

Aim To study the role of TNF-α/NF-κB signaling in matrix metalloproteinase ( MMP)-9 expres-sion induced by lipopolysaccharide ( LPS ) in airway epithelial cells, and to investigate the effects of lenti-virus mediated RNAi targeting a disintegrin and metal-loproteinase 17 ( ADAM17 ) gene on MMP-9 expression induced by LPS. Methods The ADAM17 siRNA ex-pression vector was constructed, and packaged to re-combinant lentivirus in 293T cells. The HBE4-E6/E7 cells were pretreated for 30 min by NF-κB inhibitor ( PDTC) and a recombinant human TNFR p75-Fc fu-sion protein ( Etanercept) , or infected by the recombi-nant lentivirus for 72 h, and then stimulated for 24 h by LPS or TNF-α. The release of TNF-α was detected by ELISA. The mRNA and protein levels of MMP-9 were analyzed respectively by RT-PCR and Western blot. NF-κB activity was detected by electrophoretic mobility shift assay. Results LPS and TNF-α signifi-cantly increased MMP-9 mRNA and protein expressions and the activation of NF-κB in HBE4-E6/E7 cells ( P<0. 05 ) . Etanercept and PDTC significantly inhibited MMP-9 expression and the activation of NF-κB induced by LPS ( P<0. 05 ) . Lentivirus mediated RNAi targe-ting ADAM17 significantly decreased TNF-α produc-tion, inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by LPS in HBE4-E6/E7 cells ( P <0. 05 ) . Lentivirus mediated RNAi targeting ADAM17 did not inhibit MMP-9 mRNA and protein expressions and the activation of NF-κB in-duced by TNF-α ( P>0. 05 ) . And PDTC significantly inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by TNF-α ( P <0. 05 ) . Conclusions TNF-α/NF-κB signaling partic-ipates in the regulation of MMP-9 expression induced by LPS in airway epithelial cells, and lentivirus-media-ted RNAi targeting ADAM17 plays an important role in that signaling pathway upstream by regulating TNF-αrelease.

China ; epidemiology ; Humans ; Occupational Diseases ; epidemiology ; prevention & control ; Occupational Exposure ; prevention & control ; statistics & numerical data ; Rural Population ; Transients and Migrants

Halogenated Diphenyl Ethers ; adverse effects ; Humans

AIM: To assess the impact of diabetes on corneal endothelial cells through the quantitative analysis of corneal endothelial cell morphology for patients with diabetics.
METHODS: The corneal thickness and endothelial cell morphology of 360 eyes of 299 cases were detected using full automatic corneal endothelial cell analyzer. The normal control group included 175 eyes of 148 cases, and there were 185 eyes of 151 cases for the patients with diabetes, 110 eyes of 92 cases for the non-proliferating phase group and 75 eyes of 59 cases for the proliferating phase group. The average density of central corneal endothelial cells, proportion of hexagonal cells, coefficient of variation and corneal thickness were compared among groups, and then the statistical analysis was conducted.
RESULTS: Compared with the cornea of the normal group, in the diabetes group, the coefficient of variation of corneal endothelial cells and central corneal thickness increased, while the average density of central corneal endothelial cells and proportion of hexagonal cells decreased, showing a significant difference (P<0. 05). Compared with the cornea of non-proliferating phase group, in the proliferating phase group, the density of central corneal endothelial cells decreased, the coefficient of variation of corneal endothelial cells increased, while the proportion of hexagonal cells decreased with a significant difference (P<0. 05), and the central corneal thickness increased, showing no significant difference(P>0. 05).
CONCLUSION: Compared with the cornea of normal control group, in the diabetes group, the corneal endothelial cells show abnormal morphology, which aggravates with the severity of lesions, especially for the significant changes in the coefficient of variation and the proportion of hexagonal cells. As a result, the corneal resistance to damage in patients with diabetes will decrease.

Adult ; Aged ; Carcinoma, Squamous Cell ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; pathology ; virology ; Colposcopy ; Cytodiagnosis ; DNA Probes, HPV ; DNA, Viral ; isolation & purification ; Female ; Humans ; Middle Aged ; Nucleic Acid Hybridization ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; pathology ; virology ; Vaginal Smears ; Young Adult

The purpose of this study was to construct the IgHV and IL-2 coexpressed vector. The IgHV gene fragments were obtained from the peripheral blood of patients with lymphoma, and were cloned into eukaryotic expression vector. Meanwhile, the gene fragments of IgHV linked with gene of IL-2 were inserted into pcDNA3.0 to form a fusion gene of IgHV-IL-2. Then fusion genes were transfected into COS cells by Lipofectin and the expression of IL-2 was detected by ELISA. The results showed that the IgHV/pcDNA3.0 expression vector was successfully constructed. The 3' end of IgHV was linked to IL-2 gene, and IL-2 could be correctly expressed. In conclusion, the expression vector of IgHV-IL-2 can express IL-2 correctly in COS cells.
Cancer Vaccines ; immunology ; Eukaryotic Cells ; metabolism ; Genes, Immunoglobulin ; Genetic Vectors ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Interleukin-2 ; biosynthesis ; genetics ; Lymphoma ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Vaccines, DNA ; biosynthesis ; genetics ; immunology

Objective To evaluate the clinical significance of CT angiography(CTA)in the diagnosis of arterioportal shunts(APS)associated with hepatocellular carcinoma(HCC).Methods One hundred and twenty-seven consecutive HCC patients accepted both dynamic enhancement CT and DSA examinations.The interval between CT and DSA exam was from 3 to 15 days.Based on transverse CT images in hepatic artery phase,CTA was performed for all the patients.By contrast with DSA results,the capabilities of transverse CT and transverse images combined with CTA in APS diagnosis were analyzed. Results In all 127 HCC cases,52 cases with APS were confirmed by DSA(40.94%),33 with central type of APS and 19 with peripheral type.Diagnostic sensitivity of APS based on transverse CT and combined CTA with transverse CT images were both 94.23%(49/52).However,specificity was 84.00%(63/75) and 97.33%(73/75),respectively,accuracy was 88.19%(112/127)and 96.06%(122/127),the predictive value of positive cases was 80.33%(49/61)and 96.08%(49/51),and the predictive value of negative cases was 95.45%(63/66)and 96.05%(73/76).Combined with CTA,false positive cases of 4 central type of APS and 6 peripheral type of APS were excluded which were demonstrated by transverse CT images.By contrast with DSA,the coincidence rate of the type of APS diagnosed by transverse images combined with CTA was 88.46%(46/52),including 90.91%(30/33)of central type of APS and 84.21%(16/19)of peripheral type.The supplying arteries of central type of APS were intuitively displayed by CTA in 23 cases,19 from proper hepatic artery and 4 from gastro-duodenal artery.Conclusion CTA techniques based on the dynamic enhancement CT exams could effectively promote the specificity and the accuracy of APS diagnosis.

This study was aimed to explore the expression of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 in bcr/abl fusion gene positive CML cells, and to study the effects of P210(bcr/abl) fusion protein tyrosine kinase on expression of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 mRNAs in chronic myeloid leukemia cells. The expression levels of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 mRNA were detected by semi-quantitative RT-PCR in bcr/abl negative cells, bcr/abl positive cells, and P210(bcr/abl)-Rb-C-Box positive cells. The results showed that MIP-1alpha and CCR-1 mRNAs were expressed in bcr/abl negative cells, but not in positive cells. Both MCP-1 and CCR-2 mRNA cannot be detected in both bcr/abl positive and negative cells. After inhibiting P210(bcr/abl) tyrosine kinase activity by Rb-C-Box, expressions of MIP-1alpha and CCR-1 mRNAs were restored to normal (similar to P210(bcr/abl) negative cells). It is concluded that P210(bcr/abl) fusion protein inhibits the expression of MIP-1alpha and CCR-1 in chronic myeloid leukemia cells, but does not inhibit MCP-1 and CCR-2 mRNA expressions in these leukemia cells.
Chemokine CCL2 ; biosynthesis ; genetics ; Chemokine CCL3 ; Chemokine CCL4 ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Receptors, CCR1 ; Receptors, CCR2 ; Receptors, Chemokine ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVEBone-marrow derived mesenchymal stem cells (BMSC) have the potential to differentiate into endothelial cells. The aim of the study was to investigate the induction process of BMSC by B16 melanoma cells in vitro and to analyze the role of VEGF-a in the process.

METHODSA co-culture system containing BMSC and B16 melanoma cells based on transwell indirect model was established, and the induction process of BMSC by B16 melanoma cells was studied in vitro.

RESULTSBMSC were isolated from the bone marrow of C57 mice. BMSC expressed CD105, CD90, CD73, CD44 and CD166, and acquired expressin of endothelial phenotype markers including VEGFR-1, VEGFR-2 and Factor VIII after co-culture with B16 melanoma cells for 48 hours. The expression level of VEGFR-2 would be double and Factor VIII threefold more by extending the co-culture time to 72 hours. In the co-culture system, B16 melanoma cells also up-regulated the expression of VEGF-a.

CONCLUSIONSVEGF-a plays a significant role in the differentiation of BMSC into cells of endothelial phenotype, therefore, is important to tumor angiogenesis.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Coculture Techniques ; Endothelial Cells ; cytology ; metabolism ; Factor VIII ; metabolism ; Male ; Melanoma, Experimental ; metabolism ; pathology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Pilot Projects ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism