Expression of MMP-14 and its role in bone destruction in middle ear cholesteatoma: A prospective observational study.

Cholesteatoma is a noncancerous cystic lesion caused by an abnormal growth of keratinizing squamous epithelium which is invasive and capable of destroying structures. A prospective study on the expression of membrane type1-matrix metalloproteinases (MMP-14) and its related influencing factors in middle ear cholesteatoma was conducted to fully understand the pathogenesis of cholesteatoma in the molecular level. We examined the expression of MMP-14 by immunohistochemical staining 39 middle ear cholesteatoma specimens and 10 external auditory meatus epithelial cell specimens. The cholesteatoma specimens were divided into 4 groups according to the degree of destruction of the ossicles during surgery. The associated factors affecting MMP-14 expression were analyzed using statistical methods; The positive expression of MMP-14 in the epithelium of the external auditory canal was significantly different between middle ear cholesteatoma and normal patients (P < .05); Gender, age, and the degree of hearing loss had no statistically significant effect on MMP-14 expression (P > .05); The expression of MMP-14 was positively correlated with the severity of bone destruction (R = 0.535, P < .05); MMP-14 plays an important role in the pathological development of the epithelium of cholesteatoma; MMP-14 expression in middle ear cholesteatoma tissue was not strongly correlated with the level of hearing loss, age or gender, but was positively correlated with the degree of middle ear bone destruction.

A zero-liquid-discharge scheme for vanadium extraction process by electrodialysis-based technology.

The sharp increase of demand for vanadium makes the treatment of the wastewater generated from its extraction process become an urgent problem. In this study, a hybrid process coupling the electrodialysis with the cooling crystallization is put forward for upgrading the conventional vanadium extraction process to zero discharge. Accordingly, the objective of this work lies in evaluating the feasibility of the proposed scheme on the basis of a systematic study on the influences of membrane types and operating parameters on the electrodialysis performance. The results indicate that the relative importance of osmosis and electro-osmosis to overall water transport is closely related to the applied current density. The increase in the applied current density and the decrease in the mole ratio of water and salt flux will contribute to the concentration degree. Moreover, it is worth noting that a relatively large concentration ratio can result in the remarkable decrease of current efficiency and increase of energy consumption. In general, the reclamation scheme can easily achieve the recovered water with relatively low salt content and the highly concentrated Na2SO4 solution (e.g., 300 g/L) for producing high-purity sodium sulphate crystals.

Oncogenic role of Epstein-Barr virus-encoded small RNAs (EBERs) in nasopharyngeal carcinoma.

OBJECTIVE: Epstein-Barr virus (EBV) is a causative agent of nasopharyngeal carcinoma (NPC), and EBV gene expression is considered to be closely associated with the pathogenesis of NPC. Among EBV genes expressed in NPC, EBV-encoded non-polyadenylated RNAs, termed EBERs, are the most abundant transcripts of EBV in NPC. However, the role of EBERs still remains unclear. This study was designed to investigate the relevance of EBERs to the oncogenesis of NPC. METHODS: Two types of EBERs expression vectors (EBERs-high-expression vector and EBERs-low-expression vector) were constructed and transfected into EBV-negative cells, MDCK or EBV-negative clones of NPC-KT cells. Then, malignant transformation, represented by anchor independent growth, was evaluated between the EBERs-transfected cells and EBERs-negative cells using a soft agar colony formation assay. Apoptosis was induced by serum deprivation (0.1% concentration of fetal bovine serum) and interferon-alpha (IFN-alpha) (500 U/ml) treatment. Cell viability was evaluated with a trypan blue exclusion test. The activation of cellular transcriptional factor NF-kappaB was studied with the IL-8 promoter sequence using a luciferase reporter assay. RESULTS: EBERs-high-expression vector-transfected MDCK cells showed enhanced growth ability in soft agar compared with either EBERs-low-expression vector-transfected MDCK cells or EBERs-untransfected MDCK cells. However, they did not show the acquisition of any anti-apoptotic potential against either IFN-alpha or serum deprivation. Introduction of EBERs-low-expression vector into MDCK cells did not show anchor independent growth characteristics. Neither EBV-negative NPC-KT cells nor MDCK cells transfected with EBERs-high-expression vector showed any difference from EBERs-untransfected EBV-negative NPC-KT cells. Introduction of EBERs into MDCK cells did not transactivate the IL-8 promoter, indicating that neither NF-kappaB nor AP-1 was activated by EBERs. CONCLUSION: EBERs are believed to induce the initial transformation of epithelial cells, thus contributing to the oncogenesis of NPC. Expression of abundant EBERs is considered to be critical for this transforming property of EBERs.

Epstein-Barr virus (EBV) latent membrane protein 1 induces interleukin-8 through the nuclear factor-kappa B signaling pathway in EBV-infected nasopharyngeal carcinoma cell line.

BACKGROUND/OBJECTIVES: Nasopharyngeal carcinoma (NPC) is a highly invasive and metastatic malignant tumor and is associated with Epstein-Barr virus (EBV) infection that exhibits type II latency. Angiogenesis is essential for tumor growth, invasion, and metastasis. Our previous studies have indicated that interleukin (IL)-8 was over-expressed in many NPC tissues and was found to be significantly correlated with angiogenesis by immunohistochemistry. STUDY DESIGN: In vitro design. METHODS: The influence of the EBV genome for IL-8 gene expression was studied using the EBV-genome-positive and -negative epithelial/NPC hybrid cell line NPC-KT. The EBV-positive and -negative clones were selected by polymerase chain reaction and in situ hybridization. RESULTS: EBV-positive clones expressed abundant IL-8 mRNA compared with EBV-negative clones. This result indicated that over-expression of IL-8 depended on the presence of EBV genomes in NPC-KT cells. Two encoded genes, latent membrane protein (LMP)1 and EBV-encoded small RNAs (EBERs), expressed in NPC were transfected in EBV-negative NPC-KT cells. LMP1 transactivated the IL-8 promoter, whereas EBERs did not. Moreover, the nuclear factor (NF)-kappa B binding site in the IL-8 promoter was essential for the response to LMP1, and the activator protein (AP)-1 binding site played only a partial role. CONCLUSIONS: LMP1 induces IL-8 mainly through the activation of NF-kappa B and partly through AP-1 in NPC model cell lines, NPC-KT, and this suggests that LMP1 plays an important role in the angiogenesis of NPC.

[Malignant transformation of a human oral squamous cell carcinoma cell strain OSC-19 by EBV encoded small RNAs].

OBJECTIVE: To observe the growth characteristics of the EBERs-transfected OSC (oral squamous carcinoma)-19 cells, through generated overexpression of EBERs in OSC-19 cells. METHOD: EBERs fragment were amplified by PCR from EBV-positive NPC-KT cell DNA. Using molecular biologic technique, we constructed the EBERs-expressing plasmids, which contained, 4 or 10 tandem repeats of the EBERs. The OSC-19 cells were transfected with those plasmids, respectively. We demenstrated the overexpression of EBERs in EBERs-transfected OSC-19 cells by Northern Blot analysis, and compared the growth characteristics in culture and colony formation in soft agar of Hygr- and EBERs-transfected OSC-19 cells. RESULT: 10EBERs-transfected OSC-19 cells lost cell-to-cell contacts and acquired long spindle-shape morphology in culture and formed colonies in soft agar. However, parental and Hygr-transfected OSC-19 cells scarcely did. CONCLUSION: In this study, we established 10 EBERs-transfected OSC-19 cells, which overexpressing the EBERs and showed that it has a scattering activity in culture and a role in anchorage-independent growth in soft agar. These results indicate that EBERs has a role of malignant transformation in OSC-19 cells. However, the 10 EBERs-transfected OSC-19 cell provides a useful epithelial model for studying the function of EBERs.