Shikonin down-regulates CXCR4 expression and inhibits CXCL12-induced migratory responses in colorectal carcinoma cell line SW480 / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of shikonin on the proliferation, expression of CXCR4 and the migratory responses to CXCL12 in colorectal carcinoma cell line SW480.</p><p><b>METHODS</b>The proliferation of SW480 cells was assessed by MTT assay. Cell surface expression of CXCR4 was determined by flow cytometry. The migratory ability was determined by Transwell.</p><p><b>RESULTS</b>Shikonin inhibited the proliferation of SW480 cells in time- and concentration-dependent manner. The expression rate of CXCR4 in SW480 cells was 99.1%. After application of shikonin 0.01 micromol/L, 0.1 micromol/L and 1.0 micromol/L for 24 h, the expression rate of CXCR4 decreased to 76.0%, 59.1% and 35.5% respectively (F=1098.041, P <0.001), and the CXCL12-induced SW480 cell migratory inhibition rate was 25.2%, 38.5% and 55.7% respectively (F=48.970, P <0.001).</p><p><b>CONCLUSION</b>Besides having inhibiting tumor cell proliferation effect, Shikonin may also play a role in anti-metastasis via down-regulating the expression of CXCR4 and reducing the CXCL12-induced migratory response in colorectal carcinoma cell.</p>

Changes of Toll-like receptor (TLR) 2 and TLR4 on the peripheral blood mononuclear cells in patients with chronic hepatitis B and chronic severe hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the changes of TLR2 and TLR4 on peripheral blood mononuclear cells (PBMCs) and their role in the pathogenesis of chronic hepatitis B and chronic severe hepatitis B.</p><p><b>METHODS</b>The expressions of TLR2 and TLR4 on 10000 CD14+ PBMCs were determined by flow cytometry in 30 healthy controls, in 31 patients with chronic hepatitis B and in 30 patients with chronic severe hepatitis B. The level of serum tumor necrosis factor alpha (TNF alpha) was determined by ELISA. The differences of expression of TLR2 and TLR4 on PBMCs and serum TNFalpha among the three groups of study subjects were determined by Student-t test. The correlations between TLR2, TLR4 and TNF alpha were determined by linear correlation test.</p><p><b>RESULTS</b>The values of mean fluorescence intensity (MFI) of TLR2 on PBMCs of the healthy controls, patients with chronic hepatitis B and patients with chronic severe hepatitis B groups were 21.5+/-2.7, 39.0+/-4.1, and 47.7+/-21.4; TLR4 of those groups was 2.3+/-1.1, 3.7+/-2.3, and 6.9+/-4.1. The serum TNF alpha(ng/L) of the respective groups was 53.8+/-38.1, 164.3+/-89.9, and 359.8+/-140.0. There was a gradual increase of these values from the group of healthy controls to the group of patients with chronic hepatitis B and patients with chronic severe hepatitis B. No significant positive correlations between TLR2, TLR4 and serum TNFalpha were found.</p><p><b>CONCLUSION</b>TLR2 and TLR4 may have a role in the pathogenesis of chronic hepatitis B and chronic severe hepatitis B.</p>

A close relationship between viral hepatitis B and Toll-like receptor 2 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the relationship of Toll-like receptor 2 (TLR2) and viral hepatitis B through testing the expression of TLR2 in liver tissues of patients infected with HBV and in HepG2 and HepG2.2.15 cell lines.</p><p><b>METHODS</b>Expression of TLR2 was determined by Elivision immunohistochemistry in liver tissues from patients with chronic viral hepatitis B (CHB), chronic severe hepatitis (CSH), from healthy controls and from cells of HepG2 and HepG2.2.15 hepatocellular carcinoma cell lines. Direct immunofluorescence flow cytometry was used to detect TLR2+ cell percentage and mean fluorescence intensity (MFI).</p><p><b>RESULTS</b>The intensity of TLR2 expression in liver tissues of CHB and CSH was significantly higher than that of the healthy controls (probability value less than 0.01). The positive staining was mainly located in the cytoplasm and on some cell membranes. In CHB, the intensity of TLR2 expression was positively correlated with the grade of necroinflammatory activity (r=0.597, P less than 0.01). There were significant differences of serum total bilirubin levels with different grades of positive staining of TLR2 (P less than 0.05). In liver tissues of the CHB patients, the positive staining of TLR2 was shown in small foci. MFI of TLR2 (10.7+/-2.8) and TLR2+ cell percentage (16.3%+/-7.0%) of HepG2.2.15 cells were both significantly higher than those of HepG2 cells (1.0+/-0.3, 0.4%+/-0.1%, P less than 0.01).</p><p><b>CONCLUSIONS</b>Expression of TLR2 is closely correlated with hepatitis B, especially to the grades of its necroinflammatory activity.</p>

Changes of RANTES levels in livers of patients with chronic hepatitis B: the clinical significance and the possible mechanisms / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study the relationship between intra-hepatic levels of regulated on activation, normal T-cell expressed and secreted (RANTES) and the disease severity and liver inflammatory degrees in patients with chronic hepatitis B and the possible mechanism of the changes of intra-hepatic levels of RANTES.</p><p><b>METHODS</b>The expression of RANTES of the livers was studied using immunohistochemical stainings and morphometric quantitative measurements in liver specimens from 10 normal subjects and 64 patients with chronic hepatitis B with different degrees of liver inflammation and different clinical severity. The expressions of RANTES protein and mRNA in cell line HepG2, HepG2.2.15 and HepG2 treated with 10 ng/ml TNFa at different times were quantified by ELISA and one-step RT-PCR.</p><p><b>RESULTS</b>The expression of RANTES of the livers in patients was significantly higher than that in the normal controls. Hepatic RANTES levels increased significantly and the increases were parallel to the increases of the severity of the hepatitis, from mild, moderate to severe hepatitis (the positive units were 3.7+/-1.5, 15.6+/-6.9, 24.0+/-4.0, 37.9+/-11.1, respectively) and from G0 degree to G4 degrees of liver inflammation (the positive units were 3.7+/-1.5, 15.0+/-5.7, 21.6+/-5.9, 30.3+/-8.2, 40.9+/-12.3, respectively). The expressions of RANTES protein and mRNA of HepG2.2.15 were higher than that of HepG2. RANTES protein and mRNA were induced in HepG2 by TNFa.</p><p><b>CONCLUSION</b>RANTES may have an important role in the pathogenesis of chronic hepatitis B. The elevation of hepatic RANTES may be caused by hepatitis B virus and TNFa.</p>

Differential response of developing human fetal lung explants to hyperoxia and the effects of dexamethasone on hyperoxia-exposed lungs / 中国当代儿科杂志

<p><b>OBJECTIVE</b>This study aimed to explore the effects of hyperoxia on the development of fetal lung by investigating the changes of morphological and cell proliferation induced by hyperoxia in cultured fetal lungs as well as the effects of dexamethasone on hyperoxia-exposed lungs.</p><p><b>METHODS</b>Human fetal lung explants at the pseudoglandular stage of development were cultured randomly either in normoxia (21% O2/5% CO2) or hyperoxia (95% O2/5% CO2) for 72 hrs. Dexamethasone was added into the feeding medium at the concentration of 10(-6)M. Harvested tissues were stained for pancytokeratin to identify epithelial cells, with Ki-67 as a marker of proliferation. The effects of lung morphometry were analyzed using computer assisted image analysis. The mean airway thickness, the proportion of the surface area occupied by airways, the mean airway surface area and the index of the epithelium proliferation were measured.</p><p><b>RESULTS</b>The lung architectures remained unchanged after 72 hrs normoxia culture, whereas hyperoxia culture resulted in significant dilation of airways and thinning of epithelium, with the surface area of airways of 6662 microm(2) vs 2728 microm(2) and the thickness of airways of 7.8 microm vs 8.1 microm (P < 0.05). Hyperoxia culture also resulted in an increase in the proportion of the surface area occupied by airways than normoxia culture (35.2% vs 23.4%; P < 0.05). The surface area of airways (3174 microm(2)) and the proportion of the surface area occupied by airways (23.9%) decreased significantly in hyperoxia-cultured lungs after dexamethasone administration (P < 0.05). The epithelium proliferation index in hyperoxia-cultured lungs (21.8%) was higher than that in normoxia-cultured lungs (5.1%) and dexamethasone-treated hyperoxia-cultured lungs (7.4%) (P < 0.05).</p><p><b>CONCLUSIONS</b>The exposure of pseudoglandular lungs to hyperoxia modulates the lung architecture to resemble saccular lungs with higher epithelium proliferation index. Dexamethasone may inhibit the effects induced by hyperoxia.</p>