Double low-dose computed tomography (CT) angiography of craniocervical arteries using a test bolus of diluted contrast medium and a personalized contrast protocol.

AIM: To prospectively assess the value of a test bolus of diluted contrast medium (CM) combined with a personalized contrast protocol in craniocervical computed tomography angiography (cc-CTA) with low radiation and CM doses. MATERIALS AND METHODS: Eighty-six consecutive subjects were divided into two groups at random (43 in each one): group A: 100/Sn140 kVp, filtered back-projection reconstruction, iopromide (370 mgI/ml) 50 ml; group B: 80/Sn140 kVp, iterative reconstruction, iodixanol (270 mgI/ml). In group B, the test bolus contained 27 ml of diluted CM, a personalized protocol with low-concentration CM was used for angiography, and the test bolus injection duration in angiography remained the same. Artery values over 200 Hounsfield units were considered significant. RESULTS: Image quality for all cases was found to be diagnostic. No significant differences were found in the arterial densities of the ascending aorta or basilar artery between the groups. The values of the common carotid artery, internal carotid artery, and middle cerebral artery in group B were significantly lower. The effective dose and average iodine uptake were significantly lower in group B. CONCLUSION: With double-low-dose cc-CTA, test bolus scanning based on diluted CM combined with a personalized contrast protocol can yield diagnostic-quality images and significantly reduce the radiation and CM doses.

Relationship between TLR4 and MCP2 expression levels and habitual abortion.

Habitual abortion is associated with the altered expression of multiple genes. This study was carried out to investigate the relationship between expression of Toll-like receptor 4 (TLR4) and monocyte chemotactic protein 2 (MCP2 or CCL8) and habitual abortion. This was done by detecting and comparing their relative expression in peripheral blood and placental villi of patients and healthy fertile women. Based on our previous research, 85 subjects with habitual abortion (study group) and 40 healthy fertile women (control group), who were admitted to our hospital between June 2013 and December 2014, were enrolled in this study. After these subjects signed written informed consent, peripheral blood samples and villous tissues were collected, from which the total RNA was extracted. The expression of TLR4 and MCP2 was detected with quantitative reverse transcription-polymerase chain reaction, using GAPDH as a reference control. The expression of TLR4 and MCP2 in the peripheral blood and villous tissues of the study group was significantly higher than that of the control group (P < 0.05). A positive correlation was also observed between the changes in expression levels of TLR4 and MCP2. In conclusion, TLR4 and MCP2 expression correlated with the occurrence of habitual abortion. Detecting expression changes in TLR4 and MCP2 in the peripheral blood is a feasible method for predicting the occurrence of abortion in women of child-bearing age.

Correlation of coronary artery stenosis evaluation with left heart structure and function by multi-slice computed tomography.

The aim of this study was to determine the impact of multi-slice computed tomography (MSCT) evaluation of coronary artery stenosis on left heart structure and systolic function. Coronary artery CT angiography was performed in 200 patients diagnosed with coronary heart disease, and then according to the AHA coronary artery 17-segment fractionation method, the Gensini score (GS) was determined for every narrow segment, and one-stop assessment of the correlation between left heart structure and function was performed. After the grouping of GS quartiles from low to high, there were differences between different patients with regard to LVDD, LADD, LVEDV, LVESV, MM, LVEF, and FS, while no difference in SV and CO. GS showed linear negative correlation with LVEF and FS, and linear positive correlation with LVDD, LADD, LVEDV, LVESV, and MM, while no correlation with SV and CO. That is, GS of coronary artery stenosis was negatively correlated with left ventricular systolic function and positively correlated with myocardial mass. The narrower the coronary artery, the worse the cardiac function and the higher the myocardial hypertrophy. Coronary artery stenosis was one of the important causes of the decrease in left ventricular systolic function and cardiac remodeling.

Effects of 1,25-dihydroxyvitamin D3 analogue 1,24(OH)2-22-ene-24-cyclopropyl D3 on proliferation and differentiation of a human megakaryoblastic leukemia cell line.

The novel analogue of 1,25-dihydroxyvitamin D3 (1,25(OH)2 D3), 1,24(OH)2-22-ene-24-cyclopropyl D3 (calcipotriol, MC903), exhibits similar effects on cell proliferation and cell differentiation in a newly established human megakaryoblastic leukemia cell line (HIMeg). MC903 was found to inhibit cell proliferation and induce cell differentiation in a liquid culture system at concentrations comparable to those of 1,25(OH)2 D3. Colony formation assay showed that MC903 or 1,25(OH)2 D3 markedly diminished the colony-forming ability of HIMeg cells at concentrations of 10(-6) M to 10(-10) M. Cell cycle analysis demonstrated that, as seen with 1,25(OH)2 D3, MC903 also altered the cell cycle distribution; the fraction of cells in G0 + G1 increased while those in S and G2 + M decreased. It can be concluded from these findings that 1,25(OH)2 D3 and its analogue MC903 have approximately equipotent effects on cells of megakaryoblastic lineage and are potentially useful in studying the cellular processes that are responsible for megakaryocytopoiesis.

Demonstration of vitamin D receptor expression in a human megakaryoblastic leukemia cell line: regulation of vitamin D receptor mRNA expression and responsiveness by forskolin.

We have shown earlier that 1,25-dihydroxyvitamin D3 [1,25(OH)2 D3] induces cell growth suppression and cell differentiation of a human megakaryoblastic leukemia cell line, HIMeg. However, the molecular mechanism of 1,25(OH)2 D3 action is still unknown. Prompted by this, we have searched here for the presence of 1,25(OH)2 D3 receptor (VDR) expression in HIMeg cells by reverse transcription-polymerase chain reaction (RT-PCR). The amplified product showed an identical size to the product amplified from the control human VDR cDNA and hybridized specifically with the digoxigenin-labeled human VDR cDNA fragment. As expected, VDR mRNA is also expressed in HOS-8603, a human osteosarcoma cell line. These results represent the first reported evidence that VDR mRNA is expressed in megakaryoblastic cells. In addition, the regulation of VDR mRNA expression in HIMeg cells was studied by quantitative RT-PCR. It was found that [correction of the] VDR mRNA expression in HIMeg cells could be down-regulated rapidly by 1,25(OH)2 D3 (10 nM) in a time-dependent manner, reaching a maximal reduction to about 15% of control. However, VDR mRNA expression in HOS-8603 cells was not regulated by 1,25(OH)2 D3 at any time-point tested. Treatment of HIMeg cells with forskolin (1 microM), an activator of adenylate cyclase, caused an increase in VDR mRNA levels. Similarly, VDR mRNA expression in HOS-8603 cells was also up-regulated by forskolin. Consistent with the functionality of the VDR in other target cells, we found that the up-regulation of VDR expression in HIMeg cells by forskolin was accompanied by an increased responsiveness of HIMeg cells to 1,25(OH)2 D3 even though forskolin alone had no effects. Exposure to 1,25(OH)2 D3 in combination with forskolin resulted in a much more significant inhibition of cell proliferation than to 1,25(OH)2 D3 alone. Similarly, forskolin could also augment the differentiation induced by 1,25(OH)2 D3 reflected by a more evident morphological change and a higher percentage of development of cells with multilobular nuclei. These alterations were accompanied by a loss of clonogenic capacity and a decrease in the number of cells in the S phase. These data establish that HIMeg cells express functional VDR, which served to mediate actions of its ligand on the proliferation and differentiation of these cells.

Effects of retinoic acid and dexamethasone on proliferation, differentiation, and glucocorticoid receptor expression in cultured human osteosarcoma cells.

HOS-8603 is a newly established human osteosarcoma cell line with phenotypic characteristics of osteoblasts. When these cells were grown in monolayer culture in the presence of dexamethasone (Dex) or retinoic acid (RA), there was a significant inhibition of proliferation in a concentration-dependent manner. The combined effects of Dex and RA depended upon the concentrations: at low concentrations (< 10 nM) the effects of Dex and RA were additive, whereas at high concentrations the effects were antagonistic. Anchorage-independent growth studies performed in methylcellulose culture indicated that Dex or RA inhibited colony formation by HOS-8603 cells. Treatment of HOS-8603 cells with 100 nM Dex induced alkaline phosphatase activity in a time-dependent manner, reaching a maximum of about 6.5-fold over basal levels. All these effects of Dex on HOS-8603 cells could be reversed by RU 486, a potent antiglucocorticoid. Based upon saturation of specific binding and Scatchard plot analysis, we demonstrated that a saturable, high-affinity glucocorticoid receptor (GR) existed in HOS-8603 cells, suggesting that the effects of glucocorticoids on HOS-8603 cells are mediated by the specific GR. Finally, we further investigated the homologous and heterologous regulation of GR in HOS-8603 cells. Treatment of these cells with Dex led to a time-dependent decrease in GR concentrations. This homologous GR downregulation occurred not only at the level of hormone binding but also at the level of GR mRNA. In contrast, RA was capable of increasing GR concentrations in a concentration- and time-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS)

[Induction of HSP70 mRNA in the brain and liver of rats after scalding].

Induction of hsp70 mRNA in rats after scalding was analysed using 32P labeled hsp70 cDNA. The results showed that the expression of hsp70 mRNA in brain and liver was induced markedly after scalding. Time course of the increase showed that induction took place within 5 min and subsided 24 h after scalding. In the liver, induction took place still earlier, i.e. within 1 min and lasted for more than 24 h. The induction of hsp70 mRNA in rats after scalding did not seem to be due to elevation of core temprature since the increase of temperature of rat brain and liver after scalding was not more than 1 degree C. Western blot anlyses further showed that the induction of hsp70 occured within 5 min and lasted for more than 3 h in the brain of rats after scalding. The above results demonstrated that the hsp expression in internal organs could be induced by local injuries and was not necessarily dependent upon the direct effect of stimuli in intact animals.

Effects of heat shock on glucocorticoid receptor.

The changes of glucocorticoid receptor (GR) during the heat shock response have been studied using a human osteosarcoma cell line (HOS-8603) as the model. The expression of the heat shock protein 70 (hsp70) mRNA in HOS-8603 cells has been enhanced markedly after a heat treatment at 43 degrees C for 30 min. A mild thermal pretreatment (42 degrees C for 1 h) protects the HOS-8603 cells against a subsequent heat challenge (46 degrees C). This induced thermotolerance is reflected by the increase of cell viability of HOS-8603 cells. The GR binding activity in HOS-8603 cells decreased rapidly after the heat treatment at 43 degrees C; only 42.61% of controls were detected 60 min after the heat treatment. However, there was no significant change in the dissociation constant value (Kd). These results indicate that the heat shock induce not only the heat shock mRNA expression, but also the rapid reduction in GR binding activity, suggesting that there might be a functional relationship between GR action and the heat shock response.

Identification of common gene networks responsive to radiotherapy in human cancer cells.

Identification of the genes that are differentially expressed between radiosensitive and radioresistant cancers by global gene analysis may help to elucidate the mechanisms underlying tumor radioresistance and improve the efficacy of radiotherapy. An integrated analysis was conducted using publicly available GEO datasets to detect differentially expressed genes (DEGs) between cancer cells exhibiting radioresistance and cancer cells exhibiting radiosensitivity. Gene Ontology (GO) enrichment analyses, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and protein-protein interaction (PPI) networks analysis were also performed. Five GEO datasets including 16 samples of radiosensitive cancers and radioresistant cancers were obtained. A total of 688 DEGs across these studies were identified, of which 374 were upregulated and 314 were downregulated in radioresistant cancer cell. The most significantly enriched GO terms were regulation of transcription, DNA-dependent (GO: 0006355, P=7.00E-09) for biological processes, while those for molecular functions was protein binding (GO: 0005515, P=1.01E-28), and those for cellular component was cytoplasm (GO: 0005737, P=2.81E-26). The most significantly enriched pathway in our KEGG analysis was Pathways in cancer (P=4.20E-07). PPI network analysis showed that IFIH1 (Degree=33) was selected as the most significant hub protein. This integrated analysis may help to predict responses to radiotherapy and may also provide insights into the development of individualized therapies and novel therapeutic targets.

Gamma interferon down-regulates glucocorticoid receptor expression and attenuates hormone action in a human osteosarcoma cell line.

The effect of gamma interferon (IFN) on glucocorticoid receptor (GR) expression was studied in HOS-8603 cells, a human osteogenic sarcoma cell line. Treatment of HOS-8603 cells with IFN resulted in down-regulation of GR number, with no change in the binding affinity for glucocorticoids. The maximum decrease in receptor binding was evident at 10 IU/ml IFN concentration. Time-course studies revealed that the effect reached a maximum at 36 h treatments. To clarify the molecular basis for the down-regulation of GR by IFN, change in GR mRNA levels was further investigated by RNA blot hybridization analysis. It was found that there also existed a time-dependent decrease in GR mRNA levels in HOS-8603 cells after treatment with IFN. In the presence of IFN, the inhibitory effect of glucorticoids on HOS-8603 cell proliferation was blunted. Moreover, the induction of alkaline phosphatase (AKP) activity by glucocorticoids was attenuated in response to IFN treatment. These data suggest that IFN may influence GR activity which at least partially occurs at mRNA levels, and that the decrease in receptor activity in HOS-8603 cells parallels with the decrease in glucocorticoid-mediated functional responses.

Glucocorticoid-induced growth inhibition and differentiation of a human megakaryoblastic leukemia cell line: involvement of glucocorticoid receptor.

A human megakaryoblastic leukemia cell line, HIMeg, was established recently. Previous studies have shown that retinoic acid (RA) and 1,25-dihydroxyvitamin D3 [1,25(OH)2 D3] have potent effects on the proliferation and differentiation of HIMeg cells. Recently, the effect of dexamethasone (Dex), a synthetic glucocorticoid, on HIMeg cell growth and differentiation was examined in comparison with RA and sex steroid hormones. It was observed that in a methylcellulose culture system, Dex suppressed the clonal proliferation of HIMeg cells in a dose-dependent manner. The inhibitory effects of Dex could be reversed by RU486, a potent glucocorticoid antagonist. In contrast, sex steroid hormones had little effect on the clonal proliferation of HIMeg cells. In a liquid culture system, only 2% of HIMeg cells expressed glycoprotein IIb/IIIa (GPIIb/IIIa) antigen without hormone treatment, whereas 45% and 30% of the cells expressed GPIIb/IIIa following the addition of RA and Dex, respectively. To examine the molecular basis of the hormone-induced cell differentiation, glucocorticoid receptor (GR) expression was studied by Scatchard analysis. It was shown that there existed a saturable, high-affinity GR in HIMeg cells and the GR number was altered after Dex or RA treatment of the cells, suggesting that the cellular effects of glucocorticoids on HIMeg cells were mediated by GR.

Stress-induced regulation of glucocorticoid receptor gene expression in the rat.

To investigate the mechanism of the changes in glucocorticoid receptor (GR) under various stress conditions, experiments were carried out on normal and partial adrenalectomized rats by utilizing radioligand binding assay and molecular hybridization. It was found that hepatic GR binding capacity and GR mRNA in intact rats decreased significantly after the scalding stress, but the changes in GR binding capacity is mainly owing to the decrease in GR mRNA levels, and other mechanisms at protein level are also involved. In partial adrenalectomized rats, hepatic GR binding capacity and GR mRNA were increased instead of decreased, whereas serum corticosterone did not rise in response to the scalding stress. Therefore, it may be concluded that the decrease in GR binding capacity and GR mRNA in intact rats after the scalding stress is mainly the result of the increased concentration of serum corticosterone. Our further studies did demonstrate that there exists downregulation of GR mRNA in rats by exogenous administration of hydrocortisone. It is also suggested that there might be factors that can upregulate GR and GR mRNA in intact rats after the scalding stress.

Effects of calcitriol and its analogue calcipotriol on proliferation and differentiation of human osteosarcoma cells.

The effects of steroid hormone calcitriol (Cal) and its analogue calcipotriol on human osteosarcoma cell line HOS-8603 were determined. When cells grew in monolayer culture in the presence of hormones, their proliferations were inhibited both in dose- and time-dependent manners. The cells showed marked morphologic changes after a 4-d treatment to apparently less transformed fibroblast-like ones. Anchorage-independent growth studies indicated that both Cal and calcipotriol at 10 nmol.L-1 inhibited colony formation by HOS-8603 cells. As a marker enzyme of the osteoblastic phenotype, alkaline phosphatase activity was induced in response to Cal or calcipotriol 100 nmol.L-1. These results suggested that Cal and calcipotriol play an important role in regulating growth and differentiation of HOS-8603 cells.

Transactivation specificity of glucocorticoid versus progesterone receptors. Role of functionally different interactions of transcription factors with amino- and carboxyl-terminal receptor domains.

A major unanswered question of glucocorticoid and progesterone action is how different whole cell responses arise when both of the cognate receptors can bind to, and activate, the same hormone response elements. We have documented previously that the EC(50) of agonist complexes, and the partial agonist activity of antagonist complexes, of both glucocorticoid receptors (GRs) and progesterone receptors (PRs) are modulated by increased amounts of homologous receptor and of coregulators. We now ask whether these components can differentially alter GR and PR transcriptional properties. To remove possible cell-specific differences, we have examined both receptors in the common environment of a line of mouse mammary adenocarcinoma (1470.2) cells. In order to segregate the responses that might be due to unequal nucleosome reorganization by the two receptors from those reflecting interactions with other components, we chose a transiently transfected reporter containing a simple glucocorticoid response element (i.e. GREtkLUC). No significant differences are found with elevated levels of either receptor. However, major, qualitative differences are seen with the corepressors SMRT and NCoR, which afford opposite responses with GR and PR. Studies with chimeric GR/PR receptors indicate that no one segment of PR or GR is responsible for these properties and that the composite response likely involves interactions with both the amino and carboxyl termini of receptors. Collectively, the data suggest that GR and PR induction of responsive genes in a given cell can be differentially controlled, in part, by unequal interactions of multiple receptor domains with assorted nuclear cofactors.