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.
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 observe the levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in aqueous humor of patients with macular edema secondary to central retinal vein occlusion (CRVO). Methods Forty eyes of 40 consecutive patients with macular edema secondary to CRVO (CRVO group) were enrolled in this study. The patients included 25 males and 15 females. The patient age ranged from 38 to 76 years. The control group was 20 patients with senile cataract who underwent phacoemulsification, including 10 males and 10 females. The levels of VEGF165, VEGF165b, IL-6 and MCP-1 in aqueous humor were determined by enzymelinked immunosorbent assay. The correlation of VEGF, and IL-6, and MCP-1 were analyzed. Results The median aqueous level of VEGF165, IL-6 and MCP-1 were 1089.0, 165.6, 1253.0 pg/ml respectively in CRVO group, which were higher than the control group's results (168.2, 4.7, 216.4 pg/ml respectively), the differences were statistically significant (Z=-4.549, -6.008, -5.343;P＜0.001). The VEGF165b in CRVO group and control group were 834.0, 915.9 pg/ml respectively, the difference was not statistically significant (Z=-0.207，P＞0.05). The ratio of VEGF165b to VEGF165 in CRVO group and control group were 2.71, 7.28 respectively, the difference was statistically significant (t=-3.007，P＜0.05). There was a highly positive correlation between IL-6 and VEGF in CRVO group (r=0.526,P=0.001) and also mild positive correlation in control group (r=0.425，P=0.070). No correlation between MCP-1 and VEGF was observed in both groups (CRVO group: r=0.211,P＞0.05. Control group: r=-0.019,P＞0.05). Conclusions VEGF165, IL-6 and MCP-1 levels were increased in CRVO patients while the VEGF165b was normal. The ratio between VEGF165b and VEGF165 in aqueous humor of patients with macular edema secondary to CRVO was decreased.
Objective To investigate the influence on matrix metalloproteinases (MMP) 3, 9, and 13 levels of human articular cartilage cells after blocking stromal cell derived factor 1 (SDF-1)/ chemokine receptor 4 (CXCR4) signaling pathway withAMD3100 and to define the function mechanism of AMD3100. Methods A total of 144 cartilage blocks from 12 osteoarthritis (OA) patients undergoing total knee arthroplasty (OA cartilage group) and 144 normal cartilage blocks (Mankin score of 0 or 1) from 12 patients undergoing traumatic amputation (normal cartilage group). OA cartilage group was further divided into subgroups A1, B1, and C1, and normal cartilage group into subgroups A2, B2, and C2. The cartilage tissues were cultured in DMEM solution containing 100 ng/mL SDF-1 and 1 000 nmol/L AMD3100 in subgroup A, 100 ng/mL SDF-1 and 1 000 nmol/L MAB310 in subgroup B, and 100 ng/mL SDF-1 in subgroup C, respectively. The levels of MMP-3, 9, and 13 were measured by ELISA; the expressions of MMP-3, 9, and 13mRNA were tested by RT-PCR. Results ELISA and RT-PCR results showed that the levels of MMP-3, 9, and 13 and the expressions of MMP-3, 9, and 13 mRNA were significantly lower in subgroup A than in subgroups B and C at the same time points (P lt; 0.05); the levels of MMP-3, 9, and 13 and the expressions of MMP-3, 9, and 13 mRNA were significantly higher in OA cartilage group than in normal cartilage group at the same time points (P lt; 0.05). Conclusion SDF-1 could induce overexpression and release of MMP-3, 9, and 13 in the articular cartilage through the SDF-1/CXCR4 signaling pathway; AMD3100 could reduce the mRNA expressions and secretion of MMP-3, 9, and 13 in OA cartilage by blocking the SDF-1/CXCR4 signaling pathway; but AMD3100 could not make the secretion of MMP-3, 9, and 13 return to normal levels in OA cartilage.
Objective To investigate the effects of histone modification on the expression of chemokines in alveolar epithelial typeⅡ cells ( AECⅡ) in a rat model of chronic obstructive pulmonary disease ( COPD) . Methods 20 SD rats were randomly assigned to a normal control group and a COPD group. The rat model of COPD was established by cigarette smoking. Lung histological changes were observed by HE staining. AECⅡ cells were isolated and identified by alkaline phosphatase staining and electron microscopic. The mRNA expressions of monocyte chemoattractant protein ( MCP) -1, IL-8, and macrophage inflammatory protein ( MIP) -2αwere detected by real-time quantitative PCR. The expression of histone deacetylase ( HDAC) 2 was measured by western blot. Chromatin immunoprecipitation ( ChIP) was used todetect H3 and H4 acetylation, and H4K9 methylation in the promoter region of chemokine gene. Results Compared with the control group, the mRNA expressions of MCP-1, IL-8, and MIP-2αin the COPD group increased 4. 48,3. 14, and 2. 83 times, respectively. The expression of HDAC2 protein in the COPD group wassignificantly lower than in the control group ( 0. 25 ±0. 15 vs. 0. 66 ±0. 15, P lt; 0. 05) . The expression of HDAC2 had a negative correlation with the gene expressions of IL-8, MCP-1, and MIP-2α( r = - 0. 960,- 0. 914, - 0. 928, respectively, all P lt;0. 05) . The levels of H3 and H4 acetylation were higher, and H4K9 methylation level was lower in the promoter region of chemokine gene in the COPD group compared with the control group ( all P lt; 0. 05) . Conclusions MCP-1, IL-8, and MIP-2α participate and promote the lung inflammatory response in COPD. HDAC2-mediated histone modification may play an important role in COPD inflammation.
Objective To summarize the relationships between chemokines or chemokine receptors, especially CCL19/CCL21-CCR7 and CXCL12-CXCR4 axis and occurrence and development of gastric cancer. Methods Domestic and international publications online involving the relationships between chemokines, chemokine recepotors and gastric cancer in recent years were collected and reviewed. Results By regulating the microenvironment of the growth of gastric cancer, CCL19/CCL21-CCR7 played an important role in lymph node metastasis and CXCL12-CXCR4 axis played a key role in the development of peritoneal carcinomatosis. CCR7 might function as a specific prognostic marker for lymph node metastasis of gastric cancer. Blocking the CXCL12-CXCR4 axis might be useful for the future development of a more effective therapeutic strategy for gastric cancer involved in peritoneal dissemination. Conclusions Chemokines and chemokine receptors promote the evolution of gastric cancer in variable ways, so the mechanisms of which should be comprehended to provide a theoretical basis for the future treatment. As new therapeutic targets, chemokines and chemokine receptors have a prosperity for the clinic evaluation and treatment of gastric cancer.
Objective To explore the expression of chemokine receptor CCR7 in thyroid papillary microcarcinoma tissues and the relationship with clinicopathological features. Methods The CCR7 expressions in 31 cases of thyroid papillary microcarcinoma, 34 cases of thyroid papillary carcinoma which diameter＞1cm, 34 cases of nodular goiter, and 12 cases of thyroid papillary microcarcinoma contralateral normal thyroid tissues were detected by using immunohistochemistry S-P method. Results The expression positive rates of CCR7 in thyroid papillary microcarcinoma and papillary thyroid carcinoma which diameter＞ 1cm were both 100%, the difference had not statistically significant （P＞0.05）. In nodular goiter and normal thyroid tissues, the expression positive rate of CCR7 was 64.7% and 33.3%, respectively, and compared with thyroid papillary microcarcinoma, the difference had statistically significant （P＜0.05）. There were not relations between the expression of CCR7 and patient’s gender, age, capsule invasion, and lymph node metastasis （P＞0.05）. Conclusions The CCR7 in thyroid papillary microcarcinoma and thyroid papillary carcinoma which diameter＞ 1cm are both high expressions, and have the same bionomics, both prone to cervical lymph node meta-stasis, and the radical neck dissection （central area） are both need to take.