EBNA 1 is the only antigen expressed in both Epstein-Barr virus (EBV)-infected nasopharyngeal carcinoma (NPC) and Burkitt's lymphoma cells. Previous studies showed that the mRNA of EBNA 1 in these ...two tumor tissues was initiated from a promoter located in the Bam HI F fragment (Fp) on the viral genome. Two regulatory elements located in the downstream Bam HI Q region include an EBNA 1 binding site and a positive regulatory region between the Fp and the EBNA1 binding site. This data strongly suggested that a cellular factor(s) may modulate the usage of the Fp. To locate the shortest responsible viral sequence, we constructed a series of luciferase gene and chloramphenicol acetyltransferase (CAT) gene plasmids that contained various portions of the Bam HI F/Q region. Plasmid DNA was then introduced into cells to examine the promoter activity of each construct. By this method, we identified a 186-bp fragment within the Bam HI Q region that possessed the highest activity. This promoter was designated as Qp and found to be orientation-dependent and down-regulated by EBNA 1 in both the type I BL cells and human epithelial cells. Furthermore, RNase protection assay showed that a transcription initiation site was located at nucleotide 62,416 of the EBV genome. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis further confirmed that the transcript was initiated from the Qp, and not the Fp. Therefore, our data suggested that a novel promoter, Qp, located within the Bam HI Q existed for the EBNA 1 expression in the latently infected type 1 BL cells. The biological significance of the selection of the Qp needs further investigation.
In Epstein–Barr virus (EBV)-infected BL cells, the oncogenic EBV-encoded nuclear antigen 1 (EBNA 1) gene is directed from the latent promoter Qp. Yeast one-hybrid screen analysis using the −50 to −37 ...sequence of Qp as the bait was carried out to identify transcriptional factors that may control Qp activity. Results showed that Smad4 binds the −50 to −37 sequence of Qp, indicating that this promoter is potentially regulated by TGF-β. The association of Smad4 with Qp was further confirmed by supershift of EMSA complexes using Smad4-specific antibody. The transfection of a Qp reporter construct in two EBV(+) BL cell lines, Rael and WW2, showed that Qp activity is repressed in response to the TGF-β treatment. This repression involves the interaction of a Smad3/Smad4 complex and the transcriptional repressor TGIF, as determined by cotransfection assay and coimmunoprecipitation analysis. Results suggest that TGF-β may transcriptionally repress Qp through the Smad4-binding site in human BL cells.
Background:
We assessed the therapeutic effect of acupuncture in patients with perennial allergic rhinitis. Acupuncture therapy corrects the equilibrium deviation using the bidirectional regulative ...actions in treating syndromes by inserting needles into acupoints.
Objectives:
We studied the clinical outcomes and gene expression profiles of Phadiatop (Ph)-positive (+) and -negative (−) allergic rhinitis patients who were treated with acupuncture.
Methods:
Twenty-one (21) patients with allergic rhinitis 13 Ph(+), 8 Ph(−) received 8 courses of acupuncture treatment over 4 weeks. Blood samples of the patients were collected during the course of acupuncture for global analysis of gene expression profiles by Affymetrix human U133A chips. Patients completed the rhinoconjunctivitis quality of life questionnaire (RQLQ) before and after the therapy to objectively measure the therapeutic effect of acupuncture. The gene expression profile in patients with Ph(+) and Ph(−) allergic rhinitis treated before and after acupuncture was analyzed by unsupervised and supervised clustering methods.
Results:
The results of the RQLQ and the gene expression profiles were different between the Ph(+) and Ph(−) groups after receiving treatment with acupuncture. Activity, practical problems, and nasal symptoms showed significant improvement in the Ph(+) group versus the Ph(−) group. In addition, genes involved in active immune response, differential of Treg and cell apoptosis, were different in the Ph(+) and Ph(−) groups after acupuncture treatment.
Conclusions:
Differential gene expression profiles of patients with Ph(+) and Ph(−) allergic rhinitis indicate the distinct physiologic responses after receiving acupuncture treatment in these two groups. Our results suggest that personalized medical treatment should be essential for acupuncture treatment in patients with allergic rhinitis.
Backgrounds
Imatinib mesylate (STI-571), a tyrosine kinase inhibitor, has previously been demonstrated to attenuate liver fibrogenesis through inhibition of the activation of hepatic stellate cells ...(HSCs) in CCL
4
-treated rat models.
Aims
This study aimed to further evaluate the role of STI-571 in liver regeneration.
Materials and Methods
All animals were divided into four groups, and mice were treated with or without CCL
4
and STI-571 (
n
= 6 for each group).
Results
Activated cultured HSCs in vitro with STI-571 administration showed increased apoptosis and reduced proliferation, as determined by flow cytometric analysis, 3-(4, 5-cimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay, and confocal microscopy. STI-571 treatment attenuated liver fibrosis in vivo, as was evident in the results of histology, mRNA level, and expression analysis of smooth muscle actin and type I collagen. Mice treated with STI-571 had increased liver weight ratio and the improvement in liver regeneration was compatible with the change of serum interleukin 6 levels (
p
< 0.05). Further, increased apoptosis and a reduced proliferation were observed in the CCL
4
-treated mice after STI-571 treatment based on the immunohistochemical staining of Annexin V, phosphorylated STAT3, and PCNA.
Conclusion
STI-571 treatment effectively attenuated liver fibrogenesis and improved in liver regeneration in vivo and induced apoptosis in HSCs both in vitro and in vivo.
Human dihydroorotate dehydrogenase (hDHODH) is a class-2 dihydroorotate dehydrogenase. Because it is extensively used by proliferating cells, its inhibition in autoimmune and inflammatory diseases, ...cancers, and multiple sclerosis is of substantial clinical importance. In this study, we had two aims. The first was to develop an hDHODH pharma-similarity index approach (PhSIA) using integrated molecular dynamics calculations, pharmacophore hypothesis, and comparative molecular similarity index analysis (CoMSIA) contour information techniques. The approach, for the discovery and design of novel inhibitors, was based on 25 diverse known hDHODH inhibitors. Three statistical methods were used to verify the performance of hDHODH PhSIA. Fischer's cross-validation test provided a 98% confidence level and the goodness of hit (GH) test score was 0.61. The q2, r2, and predictive r2 values were 0.55, 0.97, and 0.92, respectively, for a partial least squares validation method. In our approach, each diverse inhibitor structure could easily be aligned with contour information, and common substructures were unnecessary. For our second aim, we used the proposed approach to design 13 novel hDHODH inhibitors using a scaffold-hopping strategy. Chemical features of the approach were divided into two groups, and the Vitas-M Laboratory fragment was used to create de novo inhibitors. This approach provides a useful tool for the discovery and design of potential inhibitors of hDHODH, and does not require docking analysis; thus, our method can assist medicinal chemists in their efforts to identify novel inhibitors.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Nasal polyps (NP) are regulated by proinflammatory transcription factors such as activator protein-1 (AP-1), which comprises members of the proto-oncogene Jun and Fos protein families. The binding of ...AP-1 proteins to the 12-O-tetradecanoylphorbol-13-acetate (TPA)-response element can activate target genes and regulate many critical cellular processes. The proliferating cell nuclear antigen (PCNA) gene contains AP-1 sites, and its expression is regulated by AP-1 activity. In this study, NP and inferior turbinate (IT) were evaluated, compared with normal mucosa, to see if diffuse inflammation and active cellular proliferation exist.
A diseased group of 20 subjects and control group of 20 subjects were enrolled in this study. NP and IT were evaluated with expression of phospho-c-Jun, c-Fos, PCNA, major basic protein by immunohistochemistry, and eosinophil numbers by cell counts.
The expression of phospho-c-Jun, c-Fos, PCNA, major basic protein, and eosinophil numbers showed no significant difference in IT and NP of the same patients, but all were significantly higher in IT and NP compared with normal mucosa (P < .05).
Our result demonstrated strong evidence that diffuse mucosal inflammation and active cellular proliferation do exist in rhinosinusitis with nasal polyposis. As the degree of the disease severity increases, the difference of eosinophilic infiltration and cellular proliferation activity between NP and its adjacent mucosa decreases. An integrated anti-inflammatory treatment may be more important than surgical intervention.
Objective: Whole genome amplification (WGA) is a crucial procedure for genomic DNA analysis from limited sources, such as in forensic analysis, embryo biopsy for preimplantation genetic diagnosis, or ...needle aspiration biopsies. Several strategies for WGA have been developed for either genotyping or microarray-based comparative genome hybridization (array-CGH) during the past decade. Nevertheless, there were few studies in which various WGA methods had been performed side-by-side and results evaluated with multiple methods. Materials and Methods: Ease of performance, qualitative accuracy, and quantitative fidelity of different WGA methods, such as degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), ligation-mediated PCR (LM-PCR) and strand displacement amplification (SDA), were compared in amplifyinggenomic DNA derived from karyotype-confirmed amniocytes and the cancer cell line SAOS2. Results: Using analysis with microsatellite markers, single nucleotide polymorphism markers, and array-CGH, our results suggested that: (1) genomic DNA amplified from DOP-PCR resulted in false positive and negative results by analysis with array-CGH; (2) SDA is the easiest performance method among the three WGA methods; and (3) amplified DNA products generated by LM-PCR best reflect the original genomic DNA. Conclusion: The amplified DNA products generated by LM-PCR best reflect the original genomic DNA. Key Words: array-CGH, DOP-PCR, LM-PCR, SDA, whole genome amplification
Emerging norovirus GII.17 in Taiwan Lee, Chung-Chan; Feng, Ye; Chen, Shih-Yen ...
Clinical infectious diseases : an official publication of the Infectious Diseases Society of America,
12/2015, Letnik:
61, Številka:
11
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