Functional analysis of selenok, selenot and selenop promoters and their regulation by selenium in yellow catfish Pelteobagrus fulvidraco.

The selenok, selenot and selenop are three key selenoproteins involved in stress response. Our study, using the yellow catfish Pelteobagrus fulvidraco as the experimental animal, obtained the 1993-bp, 2000-bp and 1959-bp sequences of selenok, selenot and selenop promoters, respectively, and predicted the binding sites of several transcriptional factors on their promoters, such as Forkhead box O 4 (FoxO4), activating transcription factor 4 (ATF4), Kruppel-like factor 4 (KLF4) and nuclear factor erythroid 2-related factor 2 (NRF2). Selenium (Se) increased the activities of the selenok, selenot and selenop promoters. FoxO4 and Nrf2 can directly bind with selenok promoter and controlled selenok promoter activities positively; KLF4 and Nrf2 can directly bind with selenot promoter and controlled selenot promoter activities positively; FoxO4 and ATF4 can directly bind to selenop promoter and regulated selenop promoter activities positively. Se promoted FoxO4 and Nrf2 binding to selenok promoter, KLF4 and Nrf2 binding to selenot promoter, and FoxO4 and ATF4 binding to selenop promoter. Thus, we provide the first evidence for FoxO4 and Nrf2 bindnig elements in selenok promoter, KLF4 and Nrf2 binding elements in selenot promoter, and FoxO4 and ATF4 binding elements in selenop promoter, and offer novel insight into regulatory mechanism of these selenoproteins induced by Se.

Transcriptional Regulation and Protein Localization of Zip10, Zip13 and Zip14 Transporters of Freshwater Teleost Yellow Catfish Pelteobagrus fulvidraco Following Zn Exposure in a Heterologous HEK293T Model.

Zip family proteins are involved in the control of zinc (Zn) ion homeostasis. The present study cloned the promoters and investigated the transcription responses and protein subcellular localizations of three LIV-1 subfamily members (zip10, zip13, and zip14) from common freshwater teleost yellow catfish, Pelteobagrus fulvidraco, using in vitro cultured HEK293T model cells. The 2278 bp, 1917 bp, and 1989 bp sequences of zip10, zip13, and zip14 promoters, respectively, were subcloned into pGL3-Basic plasmid for promoter activity analysis. The pcDNA3.1 plasmid coding EGFP tagged pfZip10, pfZip13, and pfZip14 were generated for subsequent confocal microscope analysis. Several potential transcription factors' binding sites were predicted within the promoters. In vitro promoter analysis in the HEK293T cells showed that high Zn administration significantly reduced the transcriptional activities of the zip10, zip13, and zip14 promoters. The -2017 bp/-2004 bp MRE in the zip10 promoter, the -360 bp/-345 bp MRE in the zip13 promoter, and the -1457 bp/-1442 bp MRE in the zip14 promoter were functional loci that were involved in the regulation of the three zips. The -606 bp/-594 bp KLF4 binding site in the zip13 promoter was a functional locus responsible for zinc-responsive regulation of zip13. The -1383 bp/-1375 bp STAT3 binding site in the zip14 promoter was a functional locus responsible for zinc-responsive regulation of zip14. Moreover, confocal microscope analysis indicated that zinc incubation significantly reduced the fluorescence intensity of pfZip10-EGFP and pfZip14-EGFP but had no significant influence on pfZip13-EGFP fluorescence intensity. Further investigation found that pfZip10 localizes on cell membranes, pfZip14 colocalized with both cell membranes and lysosome, and pfZip13 colocalized with intracellular ER and Golgi. Our research illustrated the transcription regulation of zip10, zip13, and zip14 from P. fulvidraco under zinc administration, which provided a reference value for the mechanisms involved in Zip-family-mediated control of zinc homeostasis in vertebrates.

Dexmedetomidine-mediated protection against septic liver injury depends on TLR4/MyD88/NF-κB signaling downregulation partly via cholinergic anti-inflammatory mechanisms.

BACKGROUND: Uncontrolled inflammatory responses exacerbate the pathogenesis of septic acute liver injury (ALI), posing a lethal threat to the host. Dexmedetomidine (DEX) has been reported to possess protective properties in inflammatory conditions. This study aimed to investigate whether DEX pretreatment exhibits hepatoprotection against ALI induced by lipopolysaccharide (LPS) in rats and determine its possible molecular mechanism. METHODS: Septic ALI was induced by intravenous injection of LPS. The rats received DEX intraperitoneally 30 min before LPS administration. α-Bungarotoxin (α-BGT), a specific α7 nicotinic acetylcholine receptor (α7nAChR) antagonist, was administered intraperitoneally 1 h before LPS exposure. The role of the vagus nerve was verified by performing unilateral cervical vagotomy or sham surgery before sepsis. RESULTS: The expression of α7nAChR, toll-like receptor 4 (TLR4), high mobility group box 1 (HMGB1), and cleaved caspase-3 increased, peaking 24 h during sepsis. DEX enhanced α7nAChR activation and reduced TLR4 expression upon challenge with LPS. DEX significantly prevented LPS-induced ALI, which was associated with increased survival, the mitigation of pathological changes, the attenuation of inflammatory cytokine expression and apoptosis, and the downregulation of TLR4/MyD88/NF-κB pathway. Moreover, the hepatoprotective effect of DEX was abolished by α-BGT. Further investigation established that vagotomy, compared to sham surgery, triggered more severe pathogenic manifestations and higher proinflammatory cytokine levels. The inhibitory effects of DEX were shown in sham-operated rats but not in vagotomized rats. CONCLUSIONS: Our data highlight the pivotal function of α7nAChR and intact vagus nerves in protecting against LPS-induced ALI through inhibiting the TLR4/MyD88/NF-κB signaling pathway upon pretreatment with DEX.

[The value of diffusion tensor imaging and fiber tractography in cervical spondylotic myelopathy].

OBJECTIVE: To study the diagnostic value of diffusion tensor imaging (DTI) in cervical spondylotic myelopathy. METHODS: Twenty healthy volunteers and fifty patients with cervical spondylotic myelopathy underwent DTI in the Affiliated Hospital of Medical College of Ningbo University from January 2014 to April 2015. Healthy volunteers served as controls. Fifty patients were divided into three groups (group A , B, C) according to cervical MRI scan standard. Group A (17 cases) had only the dura mater spinalis compressed; Group B (23 cases) showed the cervical spinal cord compressed, but no high signal in it; Group C (10 cases) had the cervical spinal cord compressed with high signal in the same level. The average apparent diffusion coefficients(ADC) and fractional anisotropy (FA)values in these examinee were analyzed and all subjects were performed fiber tracking. RESULTS: There was no statistically significant differences in ADC and FA values in C2/C3, C3/C4, C4/C5, C5/C6, C6/C7 of control group (P>0.05). The average ADC and FA values in control group were (0.875 +/- 0.096) x10(3) mm2/s and 0.720 +/- 0.051, respectively; compared with group A,there was no statistically significant difference; compared with group B and C, there was significant difference; comparison among group A, B, C, there was significant differences. CONCLUSION: DTI can early and accurately quantify the changes of microstructure in cervical spondylotic myelopathy. Fiber tracking can show the damage range of spinal cord lesions.

The value of diffusion tensor imaging and fiber tractography in cervical spondylotic myelopathy / 中国骨伤

<p><b>OBJECTIVE</b>To study the diagnostic value of diffusion tensor imaging (DTI) in cervical spondylotic myelopathy.</p><p><b>METHODS</b>Twenty healthy volunteers and fifty patients with cervical spondylotic myelopathy underwent DTI in the Affiliated Hospital of Medical College of Ningbo University from January 2014 to April 2015. Healthy volunteers served as controls. Fifty patients were divided into three groups (group A , B, C) according to cervical MRI scan standard. Group A (17 cases) had only the dura mater spinalis compressed; Group B (23 cases) showed the cervical spinal cord compressed, but no high signal in it; Group C (10 cases) had the cervical spinal cord compressed with high signal in the same level. The average apparent diffusion coefficients(ADC) and fractional anisotropy (FA)values in these examinee were analyzed and all subjects were performed fiber tracking.</p><p><b>RESULTS</b>There was no statistically significant differences in ADC and FA values in C2/C3, C3/C4, C4/C5, C5/C6, C6/C7 of control group (P>0.05). The average ADC and FA values in control group were (0.875 +/- 0.096) x10(3) mm2/s and 0.720 +/- 0.051, respectively; compared with group A,there was no statistically significant difference; compared with group B and C, there was significant difference; comparison among group A, B, C, there was significant differences.</p><p><b>CONCLUSION</b>DTI can early and accurately quantify the changes of microstructure in cervical spondylotic myelopathy. Fiber tracking can show the damage range of spinal cord lesions.</p>

[Diagnostic value of 3D-indirect magnetic resonance arthrography in detecting meniscal tears of the knees].

OBJECTIVE: To explore the diagnostic value of 3D-indirectmagnetic resonance (MR) arthrography in detecting meniscustears of the knees. METHODS: Form January 2013 to January 2014, routine plain MR of the knees followed by the 3D-indirect MR arthrography was performed in 42 patients with suspected meniscal tears clinically. There were 31 males and 11 females, with an average age of 38.4 years old (ranged, 21 to 67 years old). The duration of the course ranged from 2 h to 15 d. The clinical symptom was knee pain. All the patients got subsequently arthroscopic examination or operation. The sensitivity and specificity of routine plain MR and 3D-indirect MR arthrography were compared based on the results of arthroscopic examination or operation. RESULTS: The signal intensity in the area of meniscal tears on image of 3D-indirect MR arthrography was obviously higher than that of routine plain MR. The sensitivity of 3D-indirect MR arthrography was 85.79% (87/102), while the routine plain MR was 52.94% (54/102), and the specificity improved from 67.78%(61/90) to 86.67%(78/90). The difference was statistically significant (χ2 = 25.90, P < 0.01; χ2 = 9.13, P < 0.01). CONCLUSION: In comparison with the routine plain MR findings, 3D-indirect MR arthrography demonstrates meniscus tears of the knee with the better sensitivity and specificity.

The overexpression of MRP4 is related to multidrug resistance in osteosarcoma cells.

Doxorubicin (Adriamycin, ADM) is an antimitotic drug used in the treatment of a wide range of malignant tumors, including acute leukemia, lymphoma, osteosarcoma, breast cancer, and lung cancer. Multidrug resistance-associated proteins (MRPs) are members of a superfamily of ATP-binding cassette (ABC) transporters, which can transport various molecules across extra- and intra-cellular membranes. The aim of this study was to investigate whether there was a correlation between MRP4 and primary ADM resistance in osteosarcoma cells. In this paper, we chose the human osteosarcoma cell line MG63, ADM resistant cell line MG63/DOX, and the patient's primary cell GSF-0686. We checked the ADM sensitivity and cytotoxicity of all the three cells by cell proliferation assay. The intracellular drug concentrations were measured by using LC-MS/MS. We also examined MRP4 gene expression by RT-PCR and Western Blot. We found that the intracellular ADM concentration of the parent osteosarcoma cell line MG63 was higher than the ADM resistant osteosarcoma MG63/DOX cell line or the GSF-0686 cell after ADM treatment (P < 0.05). In addition, MRP4 mRNA and protein levels in ADM resistant osteosarcoma cells were higher than in MG63 cell (P < 0.05). Taking together, this work suggests that overexpression of MRP4 may confer ADM resistance in osteosarcoma cells.

Diagnostic value of 3D-indirect magnetic resonance arthrography in detecting meniscal tears of the knees / 中国骨伤

<p><b>OBJECTIVE</b>To explore the diagnostic value of 3D-indirectmagnetic resonance (MR) arthrography in detecting meniscustears of the knees.</p><p><b>METHODS</b>Form January 2013 to January 2014, routine plain MR of the knees followed by the 3D-indirect MR arthrography was performed in 42 patients with suspected meniscal tears clinically. There were 31 males and 11 females, with an average age of 38.4 years old (ranged, 21 to 67 years old). The duration of the course ranged from 2 h to 15 d. The clinical symptom was knee pain. All the patients got subsequently arthroscopic examination or operation. The sensitivity and specificity of routine plain MR and 3D-indirect MR arthrography were compared based on the results of arthroscopic examination or operation.</p><p><b>RESULTS</b>The signal intensity in the area of meniscal tears on image of 3D-indirect MR arthrography was obviously higher than that of routine plain MR. The sensitivity of 3D-indirect MR arthrography was 85.79% (87/102), while the routine plain MR was 52.94% (54/102), and the specificity improved from 67.78%(61/90) to 86.67%(78/90). The difference was statistically significant (χ2 = 25.90, P < 0.01; χ2 = 9.13, P < 0.01).</p><p><b>CONCLUSION</b>In comparison with the routine plain MR findings, 3D-indirect MR arthrography demonstrates meniscus tears of the knee with the better sensitivity and specificity.</p>