Effect of chemokines CXCL9 and CXCL10 on bone erosion in patients with rheumatoid arthritis / 北京大学学报(医学版)

OBJECTIVE@#To detect the serum level of soluble chemokines CXCL9 and CXCL10 in patients with rheumatoid arthritis (RA), and to analyze their correlation with bone erosion, as well as the clinical significance in RA.@*METHODS@#In the study, 105 cases of RA patients, 90 osteoarthritis (OA) patients and 25 healthy controls in Peking University People's Hospital were included. All the clinical information of the patients was collected, and the serum CXCL9 and CXCL10 levels of both patients and healthy controls were measured by enzyme-linked immune sorbent assay (ELISA). CXCL9 and CXCL10 levels among different groups were compared. The correlation between serum levels with clinical/laboratory parameters and the occurrence of bone erosion in RA were analyzed. Independent sample t test, Chi square test, Mann-Whitney U test, Spearman's rank correlation and Logistic regression were used for statistical analysis.@*RESULTS@#The levels of CXCL9 and CXCL10 were significantly higher in the RA patients [250.02 (126.98, 484.29) ng/L, 108.43 (55.16, 197.17) ng/L] than in the OA patients [165.05 (75.89, 266.37) ng/L, 69.00 (33.25, 104.74) ng/L] and the health controls [79.47 (38.22, 140.63) ng/L, 55.44 (18.76, 95.86) ng/L] (all P < 0.01). Spearman's correlation analysis showed that the level of serum CXCL9 was positively correlated with swollen joints (SJC), rheumatoid factor (RF) and disease activity score 28 (DAS28) (r=0.302, 0.285, 0.289; P=0.009, 0.015, 0.013). The level of serum CXCL10 was positively correlated with tender joints (TJC), SJC, C-reactive protein (CRP), immunoglobulin (Ig) A, IgM, RF, anti-cyclic citrullinated peptide antibody (ACPA), and DAS28 (r=0.339, 0.402, 0.269, 0.266, 0.345, 0.570, 0.540, 0.364; P=0.010, 0.002, 0.043, 0.045, 0.009, < 0.001, < 0.001, 0.006). Serum CXCL9 and CXCL10 levels in the RA patients with bone erosion were extremely higher than those without bone erosion [306.84 (234.02, 460.55) ng/L vs. 149.90 (75.88, 257.72) ng/L, 153.74 (89.50, 209.59) ng/L vs. 54.53 (26.30, 83.69) ng/L, respectively] (all P < 0.01). Logistic regression analysis showed that disease duration, DAS28 and serum level of CXCL9 were correlated with bone erosion in the RA patients (P < 0.05).@*CONCLUSION@#Serum levels of CXCL9 and CXCL10 were remarkably elevated in patients with RA, and correlated with disease activities and occurrence of bone erosion. Chemokines CXCL9 and CXCL10 might be involved in the pathogenesis and bone destruction in RA.

Identification of key genes involved in nasopharyngeal carcinoma / Identificação dos principais genes envolvidos no carcinoma nasofaríngeo

Abstract Introduction: Nasopharyngeal carcinoma is the most common cancer originating from the nasopharynx. Objective: To study the mechanisms of nasopharyngeal carcinoma, we analyzed GSE12452 microarray data. Methods: GSE12452 was downloaded from the Gene Expression Omnibus database and included 31 nasopharyngeal carcinoma samples and 10 normal nasopharyngeal tissue samples. The differentially expressed genes were screened by ANOVA in the PGS package. Using the BiNGO plugin in Cytoscape and pathway enrichment analysis in the PGS package, functional and pathway enrichment analyses were performed separately to predict potential functions of the differentially expressed genes. Furthermore, Transcription factor-differentially expressed gene pairs were searched, and then the transcription factor-differentially expressed gene regulatory network was visualized using Cytoscape software. Results: A total of 487 genes were screened as differentially expressed genes between the nasopharyngeal carcinoma samples and the normal nasopharyngeal tissue samples. Enrichment analysis indicated that PTGS2 was involved in the regulation of biological process and small cell lung cancer. ZIC2 and OVOL1 may function in nasopharyngeal carcinoma through targeting significantly up-regulated genes (such as PTGS2, FN1, CXCL9 and CXCL10) in the Transcription factor-differentially expressed gene regulatory network (e.g., ZIC2→PTGS2 and OVOL1→CXCL10). Conclusion: PTGS2, FN1, CXCL9, CXCL10, ZIC2 and OVOL1 might play roles in nasopharyngeal carcinoma.

Resumo Introdução: O carcinoma nasofaríngeo é o câncer mais comum originário da nasofaringe. Objetivo: Estudar os mecanismos do câncer de nasofaringe; dados do microarray GSE12452 foram analisados. Método: GSE12452 foi obtido da base de dados Gene Expression Omnibus e inclui 31 amostras de carcinoma nasofaríngeo e 10 amostras de tecido nasofaríngeo normal. Os genes diferencialmente expressos foram analisados por ANOVA no kit PGS. Usando o plugin BiNGO no Cytoscape e análise de enriquecimento da via no kit PGS, análises de enriquecimento funcional e da via foram realizadas separadamente para prever as potenciais funções dos genes diferencialmente expressos. Além disso, os pares Fator de Transcrição - genes diferencialmente expressos foram pesquisados e em seguida a sua rede reguladora foi visualizada usando o programa Cytoscape. Resultados: Um total de 487 genes foram analisados como genes diferencialmente expressos entre as amostras de carcinoma nasofaríngeo e amostras de tecido nasofaríngeo normal. A análise de enriquecimento indicou que PTGS2 estava envolvido na regulação do processo biológico e câncer pulmonar de pequenas células. ZIC2 e OVOL1 podem funcionar no carcinoma nasofaríngeo almejando-se de maneira significativa os genes suprarregulados (como o PTGS2, FN1, CXCL9 e CXCL10) na rede reguladora de fator de transcrição - genes diferencialmente expressos (p.ex., ZIC2→PTGS2 e OVOL1→CXCL10). Conclusão: PTGS2, FN1, CXCL9, CXCL10, ZIC2 e OVOL1 podem desempenhar alguns papéis no carcinoma de nasofaringe.

IFN-stimulated genes upregulation pattern in Chronic Hepatitis C

Background: The development of effective tools for the large-scale analysis of gene expression has provided new insights into the involvement of gene networks and regular pathways in various disease processes. The chemokine receptor CXCR3 is a G protein-coupled receptor found predominantly on T cells that is activated by three ligands as follow: CXCL9 [Mig], CXCL10 [IP-10] and CXCL11 [I- TAC], and play a key role in immune and inflammatory responses by promoting recruitment and activation of different subpopulations of leukocytes. Aim of the work: The study is a logical functional approach for the development of serum markers chemokines that bind to CXC chemokine receptor 3 to determine whether they play a role in the future of immune system to clear HCV, these chemokines: CXCL9, CXCL10 and CXCL11

Patients and methods: 131 male and female patients with chronic hepatitis C virus [CHCV] infection, their age ranges between 22 and 55 years,selected from the National Hepatology and Tropical Medicine Research Institute. The included patients were divided to two groups, the first group: 80 patients were investigated for the predictive values of CXCL9,10,11 and CXCR3 chemokines in peripheral blood mononuclear cells [PBMCs], the second group were fifty one patients analyzed for the expression of surface markers on CD8+T cells. Twenty healthy individuals were included to serve as controls for each group. All the patients and controls were subjected to the following: history, clinical examination, abdominal ultrasonography and collection of blood samples for routine laboratory investigation and serological assay

Results: Chemokine CXCL9, CXCL10, CXCL11 and their receptor CXCR3 expression levels are induced in PBMCs during CHCV infection, associated with increased the expression levels of CD8+T cells in CHCV patients

Conclusion: The interaction between chemokines and their receptors is essential in recruiting HCV-specific T cells to control the infection

Recommendations: The regulation of chemokines and their receptors could be a future potential therapeutic target to decrease liver inflammation and to increase specific T cell migration to the infected liver, the blocking of chemokines and chemokine receptor engagement is a therapeutic strategy that should be explored in the near future for non-responders to current anti-HCV therapy

Differential expression of CXCL1, CXCL9 CXCL10 and CXCL12 chemokines in alopecia areata

Alopecia Areata [AA] is a non-scarring, autoimmune disorder which causes hair loss. Inflammatory reactions are involved in hair loss of the scalp and/or body. The involvement of chemokine receptors in the pathogenesis of AA is well defined among which, CXCL1 acts on neutrophils and CXCL9, CXCL10 and CXCL12 and serve as T lymphocytes recruiters. To study the serum levels of ELR+ and ELR- CXCL1, CXCL9, CXCL10 and CXCL12 in the patients suffering from AA and healthy controls. The study population of consisted of 30 patients suffering from AA and 30 healthy controls. Serum concentrations of CXCL1, CXCL9, CXCL10 and CXCL12 were measured using enzymelinked immunosorbent assay [ELISA]. Current results showed that AA patients had significantly elevated serum levels of CXCL9 and CXCL10 in comparison to controls [p<0.001]. These results also demonstrated that serum levels of CXCL1 and CXCL12 were significantly decreased in AA patients compared to control [p<0.001]. CXCL9 and CXCL10 are elevated in the AA patients and may be involved in the recruitment of T lymphocytes to the inflamed tissues. Moreover, due to the significant role played by these chemokines in angiogenesis/ angiostatis phenomenon they could be considered as useful biomarkers in AA diagnosis and therapy

Investigate circulating levels of chemokines and evaluate the correlation between these chemokines and liver function indicators in autoimmune hepatitis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>This study investigated circulation levels of chemokines (CCL2, CCL5, CXCL8, CXCL9, CXCL10) in autoimmune hepatitis(AIH) patients and evaluated the correlation between these chemokines and liver function indicators.</p><p><b>METHODS</b>A total of 5 chemokines (CCL2, CCL5, CXCL8, CXCL9, CXCL10) were measured simultaneously by cytokine beads assay(CBA) in the sera of 46 patients with AIH and 12 cases of healthy control.</p><p><b>RESULTS</b>In this study we found that serum levels of CCL2 , CXCL9 and CXCL10 in AIH patients and healthy controls were 11.79:8.39 pg/ml, 11.31:2.69 pg/ml, 15.85:4.64 pg/ml, respectively , which implied these chemokines were significantly higher in AIH patients when compared to healthy control (Z=-1.958, P=0.05; Z=-4.527, P less than 0.0001; Z=-3.84, P less than 0.0001, respectively). And circulation levels of CCL2 , CXCL8 , CXCL9 and CXCL10 in pretreatment and remission stages of patients with AIH were 29.69:11.16 pg/ml, 7.2:5.38 pg/ml, 16.02:5.47 pg/ml, 90.01:13.24 pg/ml, respectively, which showed these chemokines decreased during remission from pretreatment stage levels (t=2.985, P=0.005; Z=-2.547, P=0.0112; Z=-3.187, P=0.001; t=2.12, P=0.0015, respectively). Among AIH , CXCL8 was correlated positively with lgG(r2=0.291, P=0.0039); CXCL9 was associated positively with ALT and AST(r2=0.5324 , P less than 0.0001; r2=0.3352, P less than 0.0001); CXCL10 showed a positive correlation with ALT , AST and GGT(r2=0.9551, P less than 0.0001; r2=0.8960, P less than 0.0001; r2=0.8271, P less than 0.0001).</p><p><b>CONCLUSION</b>Serum levels of CCL2, CXCL8, CXCL9 and CXCL10 are significantly higher in patients with AIH, but decrease to levels in healthy controls after successful treatment , and circulation levels of CXCL9 and CXCL10 are associated positively with liver function indicators which can react inflammation activity of liver, all these may imply that chemokines can reflect the degree of liver inflammation and may be one of the main culprits in AIH pathological damage.</p>

Expression of cytokines in mouse hepatitis B virus X gene-transfected model / 华中科技大学学报（医学）（英德文版）

The expression profile in the mouse hepatitis B virus X (HBx)-transfected model was investigated in order to lay a foundation for further study on the implication of cytokines expression in hepatitis B virus (HBV) infection. Hydrodynamic injection method via the tail vein was used to establish the animal HBx-transfected model. By using microassay, the differential expression of gene in each group was analyzed, which was further confirmed by using real-time PCR and semi-quantitative PCR. Most of chemokine genes such as Ccl2, Ccl5, Ccl9, MIG and IP-10 were up-regulated in the HBx-transfected mouse model versus the control mice, which was coincided with the microarray results. Western blotting and immunohistochemistry were applied to detect the expression of MIG and IP-10 in the liver tissues. Simultaneously, ELISA was adopted to measure the content of IFN-γ in the liver tissues. DNA microassay revealed that the expression of 611 genes changed in HBx-transfected mice as compared with that in pCMV-tag2B-transfected mice, and most of the screened chemokines were up-regulated (including MIG and IP-10). Additionally, IFN-γ protein levels were increased by 20.7% (P<0.05) in pCMV-tag2B-HBx-transfected mice as compared with the untreated mice. IFN-γ protein levels were reduced by 53.9% (P<0.05) in pCMV-tag2B-transfected mice as compared with the untreated mice, which was consistent with the up-regulation of MIG and IP-10. It was suggested HBx transfection could induce the expression of MIG and IP-10 in the liver tissues, which might play the roles in HBV-related liver immunity and cytokines-mediated antiviral effect.

Expression and significance of cartilage oligomeric matrix protein, C-X-C motif 9 and keratin 19 in oral submucosal fibrosis / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression and localization of cartilage oligomeric matrix protein (COMP), C-X-C motif 9 (CXCL9) and keratin 19 (KRT19) in oral submucous fibrosis (OSF) and to evaluate their roles in the pathopoiesis of OSF.</p><p><b>METHODS</b>The expression and localization of COMP, CXCL9 and KRT19 were investigated in the specimens of 66 patients with oral submucous fibrosis and 14 normal controls by immunohistochemistry, and their protein and mRNA expressions were detected by Western blotting and RT-PCR.</p><p><b>RESULTS</b>COMP was overexpressed in 36 (55%) cases of OSF, and 43 (65%) cases showed positive immunoreactivity for CXCL9 protein in the cytoplasm of inflammatory cells and endothelial cells in OSF. All normal buccal mucosa tissues were stained continuously and strongly for KRT19 in the cytoplasm of basal cells, only 7 (11%) of 66 OSF samples showed faint and fragmented KRT19 staining in the cytoplasm of basal cells. RT-PCR and Western blotting results were fully consistent with the immunohistochemical data.</p><p><b>CONCLUSIONS</b>COMP, CXCL9 and KRT19 play an important role in the pathopoiesis of OSF.</p>

Inflammation markers in healthy and periodontitis patients: a preliminary data screening

Advances in diagnostic research are moving towards methods whereby the periodontal risk can be identified and quantified by objective measures using biomarkers. Patients with periodontitis may have elevated circulating levels of specific inflammatory markers that can be correlated to the severity of the disease. The purpose of this study was to evaluate whether differences in the serum levels of inflammatory biomarkers are differentially expressed in healthy and periodontitis patients. Twenty-five patients (8 healthy patients and 17 chronic periodontitis patients) were enrolled in the study. A 15 mL blood sample was used for identification of the inflammatory markers, with a human inflammatory flow cytometry multiplex assay. Among 24 assessed cytokines, only 3 (RANTES, MIG and Eotaxin) were statistically different between groups (p<0.05). In conclusion, some of the selected markers of inflammation are differentially expressed in healthy and periodontitis patients. Cytokine profile analysis may be further explored to distinguish the periodontitis patients from the ones free of disease and also to be used as a measure of risk. The present data, however, are limited and larger sample size studies are required to validate the findings of the specific biomarkers.

Avanços no diagnóstico da doença periodontal levam a métodos nos quais o risco e atividade da doença periodontal podem ser identificados e quantificados por biomarcadores. Pacientes com periodontite podem apresentar elevados níveis circulatórios de marcadores inflamatórios específicos que podem ser correlacionados com a severidade da doença. Portanto, o objetivo desse estudo foi avaliar as diferenças nos níveis séricos de biomarcadores inflamatórios em pacientes saudáveis e com doença periodontal. Foram incluídos no estudo 25 pacientes (8 saudáveis e 17 com periodontite crônica). Uma amostra de 15 mL de sangue foi obtida para identificar os marcadores inflamatórios simultaneamente utilizando Array de proteínas através de citometria de fluxo. De 24 citocinas inflamatórias analisadas, apenas 3 (RANTES, MIG e Eotaxina) apresentaram diferenças estatisticamente significantes (p<0,05) entre os dois grupos. Conclui-se que alguns marcadores inflamatórios selecionados apresentam diferença de concentração em pacientes com periodontite e saudáveis. A análise do perfil de citocinas pode ser utilizada tanto para distinguir pacientes periodontais de pacientes saudáveis, como para medir o risco à doença. Contudo, mais estudos com número maior de amostras são necessários para validar os achados sobre os biomarcadores específicos.

Increment of chemokine CXCL9/Mig in plasma correlated with acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation / 中国实验血液学杂志

To investigate the relationship between the plasma levels of chemokine CXCL9/Mig and acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The plasma levels of CXCL9/Mig of 35 patients who received all-HSCT were detected by using ELISA assay, these patients included 13 patients with grade 0-I, 12 patients with grade II and 10 patients with grade III - IV aGVHD, respectively. The four different time points including prior to allo-HSCT, one week before aGVHD onset, the plateau of aGVHD and time after completely controlled, were studied. The results showed that the plasma levels of CXCL9/Mig in the patients with serious aGVHD (grade II - IV) were significantly increased during aGVHD than those in the patients without aGVHD or with slight aGVHD (P < 0.001). It was found that CXCL9/Mig levels were significantly correlated with the severity of grade aGVHD (P < 0.001). Another important finding was that CXCL9/Mig levels obviously increased at one week before aGVHD was diagnosed. CXCL9/Mig level was not obviously correlated with CMV infection or other infectious complication (P > 0.05). It is concluded that the plasma level of CXC19/Mig significantly correlated with the severity of aGVHD and plays a critical role in pathogenesis of aGVHD, the changes in plasma level of CXCL9/Mig after allo-HSCT may be used as a valuable indicator for early diagnosis of aGVHD, finally, provide a early therapeutic approach to reduce aGVHD severity and improve the outcome for patients after allo-HSCT.

Expression profile of nasal NK/T cell lymphoma by cDNA microarray / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the changes of gene expression profile in nasal NK/T cell lymphoma.</p><p><b>METHODS</b>Total RNA was extracted from the fresh nasal NK/T cell lymphoma tissue and normal lymph node. Fluorescent labeled cDNA was obtained through synthesizing process by reverse transcription. After hybridization in the two identical microarrays consisting of 4096 genes, overexpressed or underexpressed tumor related genes were analyzed.</p><p><b>RESULTS</b>In both experimental group and control group, there were six samples. A total of 365 (8.9%) genes was found to be differentially expressed by a factor of twofold or greater in both of two identical cDNA microarrays, which included oncogenes, tumor supressor genes, cell cycle regulators, apoptotic and antiapoptotic factors, DNA transcription factors, DNA repair and recombination factors, signal transduction genes, protein translation genes, as well as a large number of metabolic genes. Thirty-seven of these genes were found to be differentially expressed by a factor of fourfold or greater. The biochemical functions of these differentially expressed genes were diverse.</p><p><b>CONCLUSION</b>This study demonstrates that many different kinds of genes are possibly involved in the initiation and progression of nasal NK/T lymphoma. cDNA microarray technique is useful in screening cancer gene expression for nasal NK/T lymphoma.</p>

Expression of chemokine monokine induced by interferon-gama in patients with chronic hepatitis B / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the expression levels of monokine induced by interferon-gama; (Mig) mRNA and its association with HBV DNA and alanine aminotransferase (ALT) in patients with chronic hepatitis B.</p><p><b>METHODS</b>The level of Mig mRNA in peripheral blood mononuclear cells (PBMCs) was dynamically detected with real-time quantitative PCR, and the ratio of chemokine/GAPDH was considered to represent the final chemokine level. The plasma level of was measured by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The mean level of Mig mRNA in PBMCs of the patients with chronic hepatitis B was 0.6883-/+0.0693, which was significantly higher than that in normal controls (P<0.001). The plasma Mig level in the patients was 609.6-/+73.8 pg/ml, also significantly higher than that in normal controls (P<0.05). In patients with chronic hepatitis B, the level of Mig mRNA in the PBMCs was significantly correlated with plasma Mig level (r=0.7157, P<0.001), and plasma Mig level was correlated with plasma ALT level (r=0.7220, P<0.001) and plasma HBV DNA level (r=0.7266, P<0.001).</p><p><b>CONCLUSION</b>Both the expression of Mig mRNA in PBMCs and plasma Mig concentration are elevated in patients with chronic hepatitis B. Mig plays an important role in migration of the inflammatory cells to the liver and mediates the development of chronic hepatitis B.</p>

Genomic Organization of ht eGene for Human Mig Chemokine / 대한면역학회지

"Mig is a gamma interferon-inducible T cell chemoattractant that is a member of the chemokine family of cytokines. In order to gain a better understanding of the molecular mechanisms that regulate expression of the Mig gene, we have characterized the Mig gene and compared its structure and regulatory sequences with that of its ciosest IP10 gene. The genomic organization of the Mig gene reveals three introns that interrupt the transcribed sequence into four functional domains with a single ""CAT""- and ""TATA""-like structure. Primer extension analysis was used to identify the transcriptional initiation site that is located 50 bp upstream to the methionine codon that begins the long open reading frame. Comparison of the intron-exon structure of this gene to the gene for IP10 establishes that both genes are interrupted in precisely the same positions within homologous codons. The similarity of the intron-exon structure of the Mig and IP10 genes further support the hypothesis that Mig and IP10 genes have evolved from a common ancestral gene by gene duplication. The 5'-flanking region of Mig gene shows no overall sequence similarity with that from its closest IP10 gene whose production is also affected by gamma interferon. However, there are regions including a sequence with similarity to the NFxB binding site, AP-1 binding site, and ISRE. The r-RF-1 binding site is well conserved from -204 to -194 from the transcription start site in the Mig gene. Given the importance of IFN-r for effective immunity in tuberculosis and induction of Mig and IP10 genes in macrophages by IFN-r, we demonstrated induction of the genes Mig and IP10 with different message levels in the THP-1 human monocytic cell lines stimulated with whole M. tuberculosis. Despite the very similarity in genomic organization and the overlap in biological activities between MIG and IP10, our data described herein further support the suggestion that these chemokines rnay role nonredundantly in vivo. Moreover, our studies done on the Mig gene should provide the structural framework for future studies and begin to dissect cis-acting DNA sequences that are critical for gene regulation mediated by cell surface receptors."