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.

Halogenated Diphenyl Ethers ; adverse effects ; Humans

Air Pollution ; prevention & control ; Epidemiologic Research Design

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

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.

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

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

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.

Recurrent spontaneous abortion (RSA) has various complicated causes, and more and more researches are focused on its etiology. Genetic factors are the most common risk factors of RSA; immune factors, infection factors, male factors and female factors play an important role; environmental pollution and some other unknown factors may also be conspirators. This article presents an overview on the possible risk factors of RSA.
Abortion, Habitual ; etiology ; Female ; Humans ; Male ; Pregnancy ; Risk Factors

OBJECTIVESTo explore the effects of p210 bcr/abl fusion gene on expression of beta1 integrin and L-selectin mRNAs in mouse chronic myeloid leukemia (CML) cells.

METHODSComparisons of beta1 integrin and L-selectin mRNA levels among p210 bcr/abl negative, p210 bcr/abl positive, and p210 bcr/abl-Rb-C-Box positive cells were undertaken by quantity RT-PCR.

RESULTSIn p210 bcr/abl positive cells, L-selectin mRNA level was decreased, but beta1 integrin mRNA expression had no change as compared to those in p210 bcr/abl negative cells. When inhibition of bcr-abl tyrosine kinase activity by Rb-C-Box, the L-selectin mRNA expression restored to normal (similar to p210 bcr/abl negative cells).

CONCLUSIONp210 bcr/abl oncoprotein inhibits expression of L-selectin mRNA, but not of beta1 integrin mRNA.

Animals ; Cell Line, Tumor ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Leukemic ; Genes, Retinoblastoma ; genetics ; Integrin beta1 ; genetics ; L-Selectin ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Mice ; RNA, Messenger ; genetics ; Transfection