Objective The observe the effects of interferon-inducible protein-10 (IP-10) on proliferation, migration and capillary tube formation of human retinal vascular endothelial cells (HREC) and human umbilical vein endothelial cells (HUVEC). Methods The chemokine receptor (CXCR3) mRNA of HREC and HUVEC were quantified by reverse transcriptase polymerase chain reaction (RT-PCR). In the presence of the different concentrations of IP-10, the difference in proliferation capacity of HREC and HUVEC were analyzed by cell counting kit-8 (CCK-8) methods. Wound scratch assay and threedimensional in vitro matrigel assay were used for measuring migration and capillary tube formation of HREC and HUVEC, respectively. Results RT-PCR revealed both HREC and HUVEC expressed CXCR3. The proliferation of HREC in the presence of IP-10 was inhibited in a dosagedependent manner (F=6.202,P＜0.05), while IP-10 showed no effect on the inhibitory rate of proliferation of HUVEC (F=1.183,P＞0.05). Wound scratch assay showed a significant reduction in the migrated distance of HREC and HUVEC under 10 ng/ml or 100 ng/ml IP-10 stimulation (F=25.373, 23.858; P＜0.05). There was no effect on the number of intact tubules formed by HREC in the presence of 10 ng/ml or 100 ng/ml IP-10. The number of intact tubules formed by HREC in the presence of 1000 ng/ml IP-10 was remarkably smaller. The difference of number of intact tubules formed by HREC among 10, 100, 1000 ng/ml IP-10 and nonintervention group was statistically significant (F=5.359,P＜0.05). Conclusion IP-10 can inhibit the proliferation, migration and capillary tube formation ability of HREC and the migration of HUVEC.
Objective To investigate the expression of chemokine receptor CXCR7 and the relation between its expression and clinicopathologic characteristics in papillary thyroid carcinoma. Method The expressions of CXCR7 in 79 cases of papillary thyroid carcinoma and their paracancerous tissues，and 33 cases of benign thyroid lesion tissues were detected by immunohistochemistry. Results The positive expression rates of CXCR7 were 0（0/79），65.8%（52/79），and 30.3%（10/33） in the paracancerous tissues，papillary thyroid carcinoma tissues，and benign thyroid lesions tissues，respectively. The positive expression rate of CXCR7 in the papillary thyroid carcinoma tissues was significantly higher than that in the paracancerous tissues （P＜0.05） or benign thyroid lesion tissues（P＜0.05）. The expression of CXCR7 was correlated with lymph node metastasis （P＜0.05）. Conclusion CXCR7 might take part in tumorigenesis，progression，and lymph node metastasis of papillary thyroid carcinoma.
Objective To detect the single nucleotide polymorphisms ( SNPs) in the upstream promoter region of chemokine like factor ( CKLF) gene and analyze their possible associations with asthma and asthma-related phenotypes. Methods Direct Sequence of the 1553bp upstream promoter region of CKLF gene was performed in 245 Chinese Han human genomic DNAs ( 119 asthmatics and 126 controls) .The frequencies of alleles, genotypes, and haplotypes were determined and the association of these SNPs with asthma were further analyzed. Results Four novel SNPs, SNP88 ( T gt; C) , SNP196 ( T gt; C) , SNP568 ( C gt;G) , and SNP1047 ( C gt; G) were found in the promoter region of CKLF. The frequency of rare allele was 0. 168 ( SNP88C) , 0. 168 ( SNP196C) , 0. 352 ( SNP568G) and 0. 167 ( SNP1047G) , respectively.Haplotypes, their frequencies and the linkage disequilibrium coefficients between SNPs were constructed.Complete linkage disequilibrium( LDs) were observed between SNP88 and SNP196, SNP88 and SNP1047,as well as SNP196 and SNP1047, respectively ( D′=1. 000, r2 = 1. 000) . SNP568 was in partial LD with the other three SNPs ( r2 = 0. 366) . No association between asthma and the SNPs was observed. Conclusions Four SNPs in the regulatory region of CKLF in Chinese Han population were firstly identified. Although no significant correlation with asthma was revealed, the SNP and haplotype information is useful for other disease association studies in the future.
Objective To explore the effect of dendritic cells (DCs) allergized by K-ras mutant peptide on expressions of chemokines CCL19, CCL22, and cytoskeletal protein fascin-1. Methods DCs were derived from peripheral blood in the presence of granuloceyte/macrophage-colony stimulating factor, interleukin (IL) -4 in vitro. The DCs were collected on day 7 after culture, and were divided into non-K-ras mutant peptide group (addition of RPMI 1604 culture solution 50 μg/ml) and K-ras mutant peptide group (addition of K-ras mutant peptide 50 μg/ml). Phenotype was identified by flow cytometry. The morphological structure was observed by scanning and transmission electron microscopies, respectively. The expressions of IL-12, CCL19, and CCL22 were tested continuously by enzyme-linked immunosorbent assay (ELISA). The expression of cytoskeletal protein fascin-1 was determined by Western blot. Results ①The expressions of CD1a, CD80, and CD86 after loading K-ras mutant peptide were higher than that before loading K-ras mutant peptide (Plt;0.01). ②The DCs with petal-like and branch-like profections after loading were observed under scanning electron microscopy; The DCs with irregular shapes, branch-like or burr-like were showed under transmission electron microscopy. ③The expressions of IL-12, CCL19, and CCL22 in the Kras mutant peptide group were higher than those in the non-K-ras mutant peptide group at different times (6, 12, 24, and 48 h) after loading Kras mutant peptide (Plt;0.01). ④The expression of fascin-1 in the K-ras mutant peptide group was also higher than that in the non-K-ras mutant peptide group (Plt;0.01). Conclusion K-ras mutant peptide can promote DC to mature and improve the expression of chemokines and cytoskeletal protein which will strengthen DC migration.
ObjectiveTo investigate the effects of naringenin on the production of chemokines and its mechanism in human bronchial epithelial (HBE) cells. MethodsHBE cells were divided into a control group, a TNF-αgroup, a low-dose naringenin group, a moderate-dose naringenin group and a high-dose naringenin group. The Naringenin groups were incubated with different doses of naringenin (10, 5 and 2.5μmol/L, respectively) for 2 h. Then the naringenin groups and the TNF-αgroup were incubated with TNF-α. After 24 h of incubation, the levels of eotaxin and RANTES were determined by ELISA method, and IκBαdegradtion was detected by Western blot method. After incubated with TNF-αfor 30 min, NF-κB DNA-binding activity was detected by EMSA method. ResultsCompared with the control group, the levels of eotaxin and RANTES were significantly increased in the HBE cells stimulated with TNF-α. Naringenin had inhibitory effects on the expression of these chemokines. Naringenin abolished IκBαdegradation and reduced the DNA-binding activity of NF-κB. ConclusionNaringenin may inhibit the production of chemokines through inhibiting NF-κB pathway.
Diabetic retinopathy (DR) is one of the most common and serious diabetic complications, which is the main cause of vision loss in adults. The specific vascular and neuropathology mechanism of DR is not clear. It has been demonstrated that Inflammatory reaction might be take effects in the development and progression of DR. Monocyte chemoattractant protein-1 (MCP-1), as an important chemokine in the inflammatory response process, promotes chemotactic and activating factors, destroys the blood-retinal barrier, causes retinal vascular disease, and activates microglia, which is related to the severity of the disease. With further research on MCP-1, it is possible to use chemokines and their receptors as target cells to control or slow down the progression of DR by reducing or inhibiting the production of MCP-1 in diabetic patients in the early stages of the disease. This study can provide new ideas and new methods about preventing and treating DR.
ObjectiveTo observe the concentration of the inflammatory cytokines in vitreous of severe proliferative diabetic retinopathy (PDR) after intravitreal ranibizumab injection (IVR). MethodsA total of 80 PDR patients (80 eyes) were enrolled in this study. The patients were randomly divided into vitrectomy group (group A) and IVR combined with vitrectomy group (group B), 40 eyes in each group. The differences of sex (χ2=0.05), age (t=0.59), duration of diabetes (t=0.36), HbA1c (t=0.13) and intraocular pressure (F=0.81) between two groups were not significant (P>0.05). The eyes in group B received 0.5 mg (0.05 ml) ranibizumab injection at 7 days before operation. The vitreous samples (0.4 ml) were obtained before operation. The concentration of vascular endothelial growth factor (VEGF), interleukin (IL)-6, IL-8, intercellular adhesion molecule-1 (ICAM-1) and connective tissue growth factor (CTGF) were measured by enzyme-linked immunosorbent assays. ResultsThe concentration of VEGF and ICAM-1 were (10.70±3.60), (224.64±90.32) pg/L in group B and (72.38±23.59), (665.61±203.34) pg/L in group A. The differences of VEGF and ICAM-1 concentration between two groups was significant (t=16.34, 12.53; P<0.001). The concentration of IL-6 and IL-8 were (210.64±80.27), (156.00±57.74) pg/L in group B and (45.78±33.82), (41.07±13.82) pg/L in group A. The differences of IL-6 and IL-8 concentration between two groups was significant (t=11.97, 12.24; P<0.001). There was no difference of CTGF concentration between two groups (t=1.39, P=0.17). The CTGF/VEGF in group B was higher than that in group A (t=14.75, P<0.001). ConclusionsOne week after IVR, the concentration of VEGF and ICAM-1 are decreased, while IL-6 and IL-8 increased. There is no obvious change in CTGF, but CTGF/VEGF is increased.
Objective To observe the relationship of serum levels of homocysteine (HCY) and chemokine C-C motifligand 2 (CCL2) with cognitive impairment in COPD patients with different degrees of emphysema. Methods Sixty-twoCOPD patients identified according to emphysema phenotype classification and admitted from January 2016 to March 2017 were recruited in the study. There were 37 cases in emphysema 1-2 grade and 25 cases in emphysema 3-4 grade. Simultaneous 30 healthy subjects undergoing physical examination were recruited as control. Montreal cognitive assessment (MoCA) scale investigation and serum HCY and CCL2 test were completed. Relationship analysis was conducted on serum HCY, CCL2 levels with cognitive impairment in the COPD patients with different degrees of emphysema. Results Compared with the 1-2 grade subgroup, the PaO2 was lower, PaCO2 was higher, the plasma HCY and CCL2 levels increased in the 3-4 grade subgroup with significant differences (all P<0.05). MoCA total score and subscores were relatively low in the COPD group with emphysema than the control group (except visuospatial ability scores in the 1-2 grade subgroup). MoCA scores were statistically lower in the 3-4 grade subgroup than those in the 1-2 grade subgroup (allP<0.05). Correlation analysis showed that HCY and CLL2 levels were negatively correlated with MoCA scores and subscores (P<0.01), and HCY and CLL2 were positively correlated (bothP<0.01). The area under the receiver operating characteristic curve of HCY and CLL2 for evaluating cognitive impairment was 0.79 and 0.97, respectively. Conclusion In patients with different degrees of emphysema phenotype, serum HCY and CCL2 levels are increased in different degree, and the degree of emphysema is closely related with cognitive dysfunction.
Objective To observe the influence of triamcinolone acetonide (TA) on the expression of pigment epitheliumderived factor (PEDF) of human retinal pigment epithelial (RPE) cells. Methods Cultured humanRPE cells (4th－6th generations) were treated with four different concentrations of TA (40, 400, 4times;103 and 4times;104 mu;g/L) for three different periods (12 or 24 or 48 hours), the levels of PEDF protein in the cell culture supernatant and cell lysates were determined by Western blot. After the initial experiment, RPE cells were treated with or without tumor necrosis factor-alpha; (TNF-alpha;, 20 ng/ml） for 24 hours, followed by TA (400 mu;g/L) treatment. The levels of PEDF and phospho-p38 mitogen activated protein kinase(p-p38MAPK) protein expression in cell culture supernatant and cell lysates were measured by Western blot. Results TAtreated RPE cells had higher PEDF expression, and 400 mu;g/L TA group had the highest effect (F＝16.98,P＜0.05). 400 mu;g/L TA treatment for one, six or 24 hours, with or without TNF-alpha; pretreatment, could all promote the PEDF expression and inhibit the p-p38MAPK protein expression (F＝16.87, 10.28; P＜0.01). TNF-alpha; pretreatment alone could inhibit PEDF protein expression and promote p-p38MAPK protein expression (F=16.87, 10.28; P＜0.01). Conclusions TA can up-regulate the expression of PEDF, and downregulate the expression of p-p38MAPK in the cultured human RPE cells.
Objective To evaluate the effects of inflammatory cytokines, including tumor necrosis factorTNF-alpha; and interleukins (IL-6 and IL-8), to the expression of pigment epithelium-derived factor (PEDF) in human retinal pigment epithelium (RPE)cells. Method Cultured primary human RPE cells were treated with 20，2，0.2 , and 0.02 ng/ml of TNF-alpha;, IL-6 and IL-8 separately. The levels of PEDF expression were determined by Western blot of the supernant after 6，12，24 and 48 hours of culture. Results PEDF secretion of RPE cells was inhibited by TNF-alpha;, IL-6 and IL-8 in a time- and dose-dependent fashion. Compared with the controls, the expression of PEDF decreased significantly in 0.02 ng/ml and 6 hours group (F＝7.14, P＜0.05）, 2.00 ng/ml and 48 hours group（F=14.05，P＜0.01) , and 20.00 ng/ml and 24 hours group（F=11.53，P＜0.01). TNF-alpha; was the most strength inhibitor (F＝14,P＜0.01）.Conclusion TNF-alpha;, IL-6, and IL-8 could suppress the expression of PEDF in the cultured human RPE cells.