Hypoxia promotes lipopolysaccharide-induced CXCL10 expression in microglia / 生理学报

This study was aimed to investigate the effect of hypoxia on lipopolysaccharide (LPS)-induced CXC-chemokine ligand-10 (CXCL10) expression and the underlying mechanism. C57BL/6J mice were randomly divided into control, hypoxia, LPS, and hypoxia combined with LPS groups. The LPS group was intraperitoneally injected with 0.5 mg/kg LPS, and the hypoxia group was placed in a hypobaric hypoxia chamber (simulated altitude of 6 000 m). The serum and hippocampal tissue samples were collected after 6 h of the treatment. The levels of CXCL10 in the serum and hippocampal tissue of mice were detected by ELISA. The microglia cell line BV2 and primary microglia were stimulated with hypoxia (1% O2) and/or LPS (100 ng/mL) for 6 h. The mRNA expression level of CXCL10 and its content in culture supernatant were detected by real-time quantitative PCR and ELISA, respectively. The phosphorylation levels of nuclear factor κB (NF-κB) signaling pathway-related proteins, p65 and IκBα, were detected by Western blot. Moreover, after NF-κB signaling pathway being blocked with a small molecular compound, PDTC, CXCL10 mRNA expression level was detected in the BV2 cells. The results showed that in the LPS-induced mouse inflammatory model, hypoxia treatment could promote LPS-induced up-regulation of CXCL10 in both serum and hippocampus. Compared with the cells treated with LPS alone, the expression of CXCL10 mRNA and the content of CXCL10 in the culture supernatant of BV2 cells treated with hypoxia combined with LPS were significantly increased. The CXCL10 mRNA level of primary microglial cells treated with hypoxia combined with LPS was significantly up-regulated. Compared with the cells treated with hypoxia or LPS alone, the phosphorylation levels of p65 and IκBα in the BV2 cells treated with hypoxia combined with LPS were significantly increased. PDTC blocked the induction of CXCL10 gene expression by LPS in the BV2 cells. These results suggest that hypoxia promotes LPS-induced expression of CXCL10 in both animal and cell models, and NF-κB signaling pathway plays an important role in this process.

Impact of edaravone on serum CXC chemokine ligand-13 levels and perioperative neurocognitive disorders in elderly patients with hip replacement / 中华医学杂志(英文版)

BACKGROUND@#Perioperative neurocognitive disorders (PND) are a series of severe complications in the perioperative and anesthetic periods with a decline in memory, execution ability, and information processing speed as the primary clinical manifestation. This study aimed to evaluate the impact of edaravone (EDA) on PND and peripheral blood C-X-C motif chemokine ligand 13 (CXCL13) levels in elderly patients with hip replacement.@*METHODS@#A total of 160 elderly patients undergoing hip arthroplasty in Affiliated Dongguan People's Hospital of Southern Medical University (from March 2016 to March 2018) were randomly and double-blindly categorized into an EDA group and a control group (CON). Group EDA was administered intravenously EDA 30 min before surgery, and group CON was administered intravenously saline. The cognitive function of the two groups was evaluated 1-day before the operation and at 1 and 12 months after surgery, and the incidence of post-operative delirium was tested on days 1, 3, and 7 after surgery using the Chinese version of the confusion assessment method. Serum CXCL13 and interleukin (IL)-6 concentrations were measured before anesthesia, during surgery (30 min after skin incision), and on days 1, 3, and 7 after surgery. The continuous variables in accordance with normal distribution were tested using the Student's t test, the continuous variables without normal distribution using the Mann-Whitney U test, and categorical variables by the χ2 test or Fisher exact test.@*RESULTS@#The incidence of post-operative delirium within 7 days after surgery was significantly higher in group CON than that in group EDA (31.3% vs. 15.0%, t = -5.6, P < 0.001). The modified telephone interview for cognitive status and activities of daily life scores were significantly higher in the group EDA than those in the group CON at 1 month (39.63 ± 4.35 vs. 33.63 ± 5.81, t = -2.13, P < 0.05 and 74.3 ± 12.6 vs. 61.2 ± 13.1, t = -1.69, P < 0.05) and 12 months (40.13 ± 5.93 vs. 34.13 ± 5.36, t = -3.37, P < 0.05 and 79.6 ± 11.7 vs. 65.6 ± 16.6, t = -2.08, P < 0.05) after surgery; and the incidence of neurocognitive dysfunction was significantly lower in the group EDA than that in the group CON (P < 0.05). Serum CXCL13 and IL-6 concentrations were significantly lower in the group EDA than those in the group CON during and after surgery (P < 0.05).@*CONCLUSION@#EDA can significantly reduce the serum concentrations of CXCL13 and IL-6 and improve the PND of patients.

Ischemic preconditioning modifies mortality and inflammatory response

PURPOSE: To evaluate the effect of ischemic preconditioning on mortality, inflammatory mediators and oxidative stress after intestinal ischemia and reperfusion. METHODS: Male Wistar rats were allocated according to the period of ischemia with or without ischemic preconditioning which consist on clamping the superior mesenteric artery for 10 minutes followed by reperfusion for 10 minutes before the sustained ischemia period. Mortality was assessed in Phase 1 study, and the CINC-1, CINC-2 and MDA levels in the lungs were analyzed in Phase 2. RESULTS: Mortality was lower in the ischemic preconditioning group subjected to 90 minutes of ischemia compared to the group without ischemic preconditioning (I-90: 50% and IPC-90: 15%, p=0.018), and it was lower in the ischemic preconditioning group as a whole compared to the groups without ischemic preconditioning (IPC-14% and I=30%, p=0.006). Lower levels of MDA, CINC-1, and CINC-2 were observed in the animals that were subjected to ischemic preconditioning compared to the animals that were not (MDA: I-45=1.23 nmol/mg protein, and IPC-45=0.62 nmol/mg protein, p=0.0333; CINC-1: I-45=0.82 ng/mL and IPC-45=0.67 ng/mL, p=0.041; CINC-2: I-45=0.52 ng/mL and IPC-45=0.35 ng/mL, p=0.032). CONCLUSION: Ischemic preconditioning reduces mortality, inflammatory process and oxidative stress in rats subjected to intestinal ischemia and reperfusion.

Expression of chemokine CXCL14 in primary osteosarcoma and its association with prognosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of CXCL14 in human osteosarcoma cell lines and tissues and investigate its association with the prognosis of the patients.</p><p><b>METHODS</b>RT-PCR, enzyme-linked immunosorbent assay (ELISA) and real-time PCR were used to detect the expression of CXCL14 in 4 osteosarcoma cell lines and in 40 pairs of osteosarcoma tissues and adjacent muscular tissues. CCK8 assay and colony formation assay was used to assess the effect of CXCL14 suppression mediated by two specific siRNAs on the proliferation of U2OS osteosarcoma cells. Immunohistochemistry was performed to detect the expression of CXCL14 in 58 osteosarcoma tissues, and Kaplan-Meier method and log-rank test were performed for survival analysis of the patients.</p><p><b>RESULTS</b>Significant up-regulation of CXCL14 expression was found in the osteosarcoma cell lines and in osteosarcoma tissues compared with the adjacent muscles (P<0.01). In U2OS cell, suppression of CXCL14 expression by siRNA significantly inhibited the cell proliferation (P<0.01) and colony formation rate (P<0.05). Kaplan-Meier survival analysis indicated that patients with high CXCL14 expression had worse prognosis than those with low CXCL14 expression (P=0.02).</p><p><b>CONCLUSION</b>CXCL14 is up-regulated in both osteosarcoma cell lines and primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high CXCL14 expression in osteosarcoma tissues is associated with a poor prognosis, suggesting the that CXCL14 serve as a potential therapeutic target for osteosarcoma.</p>

Expression of the CXCL12/CXCR4 and CXCL16/CXCR6 axes in cervical intraepithelial neoplasia and cervical cancer / 癌症

The chemokine CXCL12 is highly expressed in gynecologic tumors and is widely known to play a biologically relevant role in tumor growth and spread. Recent evidence suggests that CXCL16, a novel chemokine, is overexpressed in inflammation-associated tumors and mediates pro-tumorigenic effects of inflammation in prostate cancer. We therefore analyzed the expression of CXCL12 and CXCL16 and their respective receptors CXCR4 and CXCR6 in cervical intraepithelial neoplasia (CIN) and cervical cancer and further assessed their association with clinicopathologic features and outcomes. Tissue chip technology and immunohistochemistry were used to analyze the expression of CXCL12, CXCR4, CXCL16, and CXCR6 in healthy cervical tissue (21 cases), CIN (65 cases), and cervical carcinoma (60 cases). The association of protein expression with clinicopathologic features and overall survival was analyzed. These four proteins were clearly detected in membrane and cytoplasm of neoplastic epithelial cells, and their distribution and intensity of expression increased as neoplastic lesions progressed through CIN1, CIN2, and CIN3 to invasive cancer. Furthermore, the expression of CXCR4 was associated significantly with the histologic grade of cervical carcinoma, whereas the expression of CXCR6 was associated significantly with lymph node metastasis. In Kaplan-Meier analysis, patients with high CXCR6 expression had significantly shorter overall survival than did those with low CXCR6 expression. The elevated co-expression levels of CXCL12/CXCR4 and CXCL16/CXCR6 in CIN and cervical carcinoma suggest a durative process in cervical carcinoma development. Moreover, CXCR6 may be useful as a biomarker and a valuable prognostic factor for cervical cancer.

Influência de materiais odontológicos na capacidade de resposta de fibroblastos cultivados de polpa dental humana / Influence of dental materials on the response capability of cultured fibroblasts from human dental pulp

O presente trabalho tem como objetivo investigar a influência de materiais utilizados na prática odontológica (Single Bond, HEMA, Vitrebond, Ketac Molar e Dycal) na resposta inflamatória de fibroblastos cultivados de polpa dental humana de dentes permanentes em relação à expressão e produção de mediadores da inflamação. As culturas primárias de fibroblastos foram estabelecidas a partir do tecido pulpar de terceiros molares hígidos. Após a quarta passagem, os fibroblastos foram estimulados pelos materiais e pelos materiais seguidos por LPS de E. coli pelos tempos de 6 e 24 horas. Os testes utilizados foram: MTT, Trypan Blue, Análise de Griess, PCR quantitativo e ELISA. Os dados foram analisados estatisticamente aplicando-se o teste ANOVA a 1 critério e pós-teste de Tukey e ANOVA a 2 critérios e teste de correção de Bonferroni (p<0,05). Os materiais SB10 (Single Bond 1:100) e DY (Dycal) afetaram a viabilidade celular com diminuição do metabolismo. Os materiais SB1 (Single Bond 1:1.000), SB10 (Single Bond 1:100) e VB (Vitrebond) seguidos de LPS de E. coli diminuíram o metabolismo celular de maneira estatisticamente significativa. Os níveis de óxido nítrico produzidos foram diminuídos quando os fibroblastos foram estimulados pelo KM (Ketac Molar). A expressão gênica para pró-colágeno tipo I foi diminuída quando os fibroblastos foram estimulados pelos materiais SB10 (Single Bond 1:100), SB (Single Bond polimerizado) e DY (Dycal). Para o SDF-1_/CXCL12 houve um aumento da expressão para o grupo estimulado apenas por LPS de E. coli, SB10 (Single Bond 1:100) e DY (Dycal). Para o IL-6 notou-se uma diminuição significativa para o grupo estimulado por H1000 (HEMA 1000 nM) e um aumento para o grupo SB10 (Single Bond 1:100). A expressão gênica de IL- 8/CXCL8 diminuiu para os fibroblastos estimulados pelas três concentrações de HEMA e de Single Bond, VB (Vitrebond) e DY (Dycal) no período de 6 horas e houve um aumento para os materiais SB10 (Single Bond 1:100) e VB...

The aim of the present study is to investigate the influence of dental materials (Single Bond, HEMA, Vitrebond, Ketac Molar e Dycal) in the inflammatory response of human dental pulp fibroblasts from permanent teeth in relation to inflammatory mediators expression. and production. Primary cultures were established from third molars pulp tissue. After the fourth passage, the fibroblasts were stimulated only by materials and also by the materials followed by LPS from E. coli for 6 and 24 hours. Data were statistically analyzed using Oneway ANOVA and Tukey post-test and Two-way ANOVA followed by Bonferroni post-test (p<0.05). SB10 (Single Bond 1: 100) and DY (Dycal) affected cell viability and consequently decreased cell metabolism. SB1 (Single Bond 1:1,000), SB10 (Single Bond 1:100) and VB (Vitrebond) followed by LPS E. coli decreased cell metabolism. Nitric oxide levels were reduced when fibroblasts were stimulated by KM (Ketac Molar). Pro-collagen type I expression was reduced when fibroblasts were stimulated by SB10 (Single Bond 1:100), SB (polymerized Single Bond) and DY (Dycal). SDF-1_/CXCL12 expression was increased for the group stimulated only by LPS from E. coli, SB10 (Single Bond 1:100) and DY (Dycal). IL-6 expression had a significant decrease in the group stimulated by H1000 (HEMA 1000 nM) and an increase for SB10 (Single Bond 1:100) group. The expression of IL-8/CXCL8 decreased when fibroblasts were stimulated by the three concentrations of HEMA and of Single Bond, VB (Vitrebond) and DY (Dycal) at 6 hours and increased for SB10 (Single Bond 1:100) and VB (Vitrebond) at 24 hours. There was decrease in SDF-1_/CXCL12 production for the three concentrations of HEMA and DY (Dycal) and a declining trend for the other materials tested. The production of IL-6 was increased by VB (Vitrebond) and KM (Ketac Molar). The production of IL-8/CXCL8 increased by SB1 (Single Bond 1:1,000), VB...

Construction of SDF-1P2G54, a specific antagonist of CXCR4 / 南方医科大学学报

<p><b>OBJECTIVE</b>To obtain a specific antagonist of CXCR4, SDF-1P2G54 by mutating SDF-1 second proline (P) into glycin (G) and removing the α-helix of its C-terminal.</p><p><b>METHODS</b>SDF-1p2g54 gene amplified by PCR was inserted into the vector pET-30a (+) and transformed into Escherichia coli (E. coli) strain BL21. After IPTG induction of E. coli, the expressed recombinant protein was purified with nickel-affinity chromatography column under denaturing conditions and refolded with gradient dilution and ultra-filtration. The chemotactic effect of SDF-1P2G54 on Jurkat cells and its antagonistic effect against SDF-1 were determined by transwell assay; flow cytometry was used to assay the ability of SDF-1P2G54 to induce calcium influx and CXCR4 internalization in MOLT4 cells.</p><p><b>RESULTS</b>The recombinant protein SDF-1P2G54 completely lost the functions to activate CXCR4 or to induce transmembrane migration of Jurkat cells and calcium influx in MOLT4 cells, but maintained a high affinity to CXCR4. SDF-1P2G54 effectively inhibited the chemotactic effect of wild-type SDF-1 to Jurkat cells, and induced rapid CXCR4 internalization in MOLT4 cells.</p><p><b>CONCLUSION</b>SDF-1P2G54 is a new antagonist of CXCR4 with a potential value as an effective inhibitor of HIV-1 infection, cancer metastasis or other major diseases.</p>

Azithromycin inhibits neutrophil accumulation in airways by affecting interleukin-17 downstream signals / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Azithromycin can reduce neutrophil accumulation in neutrophilic pulmonary diseases. However, the precise mechanism behind this action remains unknown. Our experiment assessed whether azithromycin inhibits neutrophil accumulation in the airways by affecting interleukin-17 (IL-17) downstream signals.</p><p><b>METHODS</b>Mice were pretreated with azithromycin before murine IL-17A (mIL-17) stimulation. After the mIL-17 stimulation, the levels of six neutrophil-mobilizing cytokines were determined by enzyme-linked immunosorbent assay (ELISA) tests in bronchoalveolar lavage (BAL) fluid; IL-6, CXC chemokine ligand-1 (CXCL-1), CXCL-5, macrophage inflammatory protein-2 (MIP-2), granulocyte colony-stimulating factor (G-CSF), and granulocyte macrophage colony-stimulating factor (GM-CSF). The number of neutrophils in BAL fluid were evaluated by cytospin preparations.</p><p><b>RESULTS</b>(1) Azithromycin pretreatment significantly inhibited both the release of three neutrophil-mobilizing cytokines (MIP-2, CXCL-5 and GM-CSF) and the accumulation of neutrophils in airways caused by mIL-17 stimulation. (2) The levels of three neutrophil-mobilizing cytokines (IL-6, MIP-2 and GM-CSF) were positively correlated with the numbers of neutrophil in BAL fluid.</p><p><b>CONCLUSIONS</b>Azithromycin can inhibit neutrophil accumulation in the airways by affecting IL-17 downstream signals. This finding suggests that macrolide antibiotic application might be useful in prevention of neutrophilic pulmonary diseases characterized by high levels of IL-17.</p>

Influência de materiais odontológicos na capacidade de resposta de fibroblastos cultivados de polpa dental humana / Influence of dental materials on the response capability of cultured fibroblasts from human dental pulp

O presente trabalho tem como objetivo investigar a influência de materiais utilizados na prática odontológica (Single Bond, HEMA, Vitrebond, Ketac Molar e Dycal) na resposta inflamatória de fibroblastos cultivados de polpa dental humana de dentes permanentes em relação à expressão e produção de mediadores da inflamação. As culturas primárias de fibroblastos foram estabelecidas a partir do tecido pulpar de terceiros molares hígidos. Após a quarta passagem, os fibroblastos foram estimulados pelos materiais e pelos materiais seguidos por LPS de E. coli pelos tempos de 6 e 24 horas. Os testes utilizados foram: MTT, Trypan Blue, Análise de Griess, PCR quantitativo e ELISA. Os dados foram analisados estatisticamente aplicando-se o teste ANOVA a 1 critério e pós-teste de Tukey e ANOVA a 2 critérios e teste de correção de Bonferroni (p<0,05). Os materiais SB10 (Single Bond 1:100) e DY (Dycal) afetaram a viabilidade celular com diminuição do metabolismo. Os materiais SB1 (Single Bond 1:1.000), SB10 (Single Bond 1:100) e VB (Vitrebond) seguidos de LPS de E. coli diminuíram o metabolismo celular de maneira estatisticamente significativa. Os níveis de óxido nítrico produzidos foram diminuídos quando os fibroblastos foram estimulados pelo KM (Ketac Molar). A expressão gênica para pró-colágeno tipo I foi diminuída quando os fibroblastos foram estimulados pelos materiais SB10 (Single Bond 1:100), SB (Single Bond polimerizado) e DY (Dycal). Para o SDF-1_/CXCL12 houve um aumento da expressão para o grupo estimulado apenas por LPS de E. coli, SB10 (Single Bond 1:100) e DY (Dycal). Para o IL-6 notou-se uma diminuição significativa para o grupo estimulado por H1000 (HEMA 1000 nM) e um aumento para o grupo SB10 (Single Bond 1:100). A expressão gênica de IL- 8/CXCL8 diminuiu para os fibroblastos estimulados pelas três concentrações de HEMA e de Single Bond, VB (Vitrebond) e DY (Dycal) no período de 6 horas e houve um aumento para os materiais SB10 (Single Bond 1:100) e VB...

The aim of the present study is to investigate the influence of dental materials (Single Bond, HEMA, Vitrebond, Ketac Molar e Dycal) in the inflammatory response of human dental pulp fibroblasts from permanent teeth in relation to inflammatory mediators expression. and production. Primary cultures were established from third molars pulp tissue. After the fourth passage, the fibroblasts were stimulated only by materials and also by the materials followed by LPS from E. coli for 6 and 24 hours. Data were statistically analyzed using Oneway ANOVA and Tukey post-test and Two-way ANOVA followed by Bonferroni post-test (p<0.05). SB10 (Single Bond 1: 100) and DY (Dycal) affected cell viability and consequently decreased cell metabolism. SB1 (Single Bond 1:1,000), SB10 (Single Bond 1:100) and VB (Vitrebond) followed by LPS E. coli decreased cell metabolism. Nitric oxide levels were reduced when fibroblasts were stimulated by KM (Ketac Molar). Pro-collagen type I expression was reduced when fibroblasts were stimulated by SB10 (Single Bond 1:100), SB (polymerized Single Bond) and DY (Dycal). SDF-1_/CXCL12 expression was increased for the group stimulated only by LPS from E. coli, SB10 (Single Bond 1:100) and DY (Dycal). IL-6 expression had a significant decrease in the group stimulated by H1000 (HEMA 1000 nM) and an increase for SB10 (Single Bond 1:100) group. The expression of IL-8/CXCL8 decreased when fibroblasts were stimulated by the three concentrations of HEMA and of Single Bond, VB (Vitrebond) and DY (Dycal) at 6 hours and increased for SB10 (Single Bond 1:100) and VB (Vitrebond) at 24 hours. There was decrease in SDF-1_/CXCL12 production for the three concentrations of HEMA and DY (Dycal) and a declining trend for the other materials tested. The production of IL-6 was increased by VB (Vitrebond) and KM (Ketac Molar). The production of IL-8/CXCL8 increased by SB1 (Single Bond 1:1,000), VB...

Chemokine axes CXCL12/CXCR4 and CXCL16/CXCR6 correlate with lymph node metastasis in epithelial ovarian carcinoma / 癌症

Recent evidence suggests that the chemokine axis of CXC chemokine ligand-12 and its receptor CXC chemokine receptor-4 (CXCL12/CXCR4) is highly expressed in gynecological tumors and the axis of CXC chemokine ligand-16 and CXC chemokine receptor-6 (CXCL16/CXCR6) is overexpressed in inflammation-associated tumors. This study aimed to determine the relationship between CXCL12/CXCR4, CXCL16/CXCR6 and ovarian carcinoma's clinicopathologic features and prognosis. Accordingly, the expression of these proteins in ovarian tissues was detected by tissue microarray and immunohistochemistry. The expressions of CXCL12/CXCR4 and CXCL16/CXCR6 were significantly higher in epithelial ovarian carcinomas than in normal epithelial ovarian tissues or benign epithelial ovarian tumors. The expression of chemokines CXCL12 and CXCL16 were positively correlated with their receptors CXCR4 and CXCR6 in ovarian carcinoma, respectively (r = 0.300, P < 0.05; r = 0.395, P < 0.05). Moreover, the expression of CXCL12 was related to the occurrence of ascites (Χ² = 4.76, P < 0.05), the expression of CXCR4 was significantly related to lymph node metastasis (Χ(2) = 4.37, P < 0.05), the expression of CXCR6 was significantly related to lymph node metastasis (Χ² = 7.43, P < 0.05) and histological type (Χ² = 33.48, P < 0.05). In univariate analysis, the expression of CXCR4 and CXCL16 significantly correlated with reduced median survival (Χ² = 4.67, P < 0.05; Χ² = 4.48, P < 0.05). Therefore, we conclude that the chemokine axes CXCL12/CXCR4 and CXCL16/CXCR6 may play important roles in the growth, proliferation, invasion, and metastasis of epithelial ovarian carcinoma.

Prognostic Implications of the Expression of CXCL16 in Breast Carcinoma

BACKGROUND: Of the many prognostic factors for breast cancer, the relationship between an infiltration of inflammatory cells and the prognosis is debatable. Of the chemokines affecting cancer's inflammatory reactions, chemokine (C-X-C motif) ligand 16 (CXCL16) has attracted attention for its prognostic value in many cancers, including colorectal cancer and renal cell carcinoma. But the situation for breast carcinoma is unknown. The aim of this study was to examine the relationship between the prognostic factors and the CXCL16 expression in patients with breast carcinoma. METHODS: The patients (n=106) diagnosed with invasive ductal cancer of the breast were enrolled. We reviewed the clinicopathological factors of these patients, hematoxylin and eosin stains were prepared and estrogen receptor (ER), human epidermal growth factor receptor 2 (HER2/neu) and CXCL16 immunostaining was performed. RESULTS: The ER expression was significantly correlated with age and inflammation. A CXCL16 expression was noted in 81.1% of the cases. No association was evident between a CXCL16 expression and any other parameter, including the survival rate. Multivariate analysis did not implicate ER, HER2/neu or CXCL16 as an independent prognostic factor, but the tumor size was independent predictive factor for the patient outcome. CONCLUSIONS: An inflammatory reaction mediated by CXCL16 is not associated with the prognosis of breast cancer or any clinicopathological factors.

The expression of CEACAM-1 and CXCL-14 in infantile hemangioma / 中华整形外科杂志

<p><b>OBJECTIVE</b>To examine the expression of CEACAM-land CXCL-14 in the different stages of infantile hemangioma and to explore the role of CEACAM-1 and CXCL-14 in the occurrence and development of infantile hemangioma.</p><p><b>METHODS</b>The expression of CEACAM-1 and CXCL-14 was detected by immunohistochemical technique and Western Blot in cases of proliferating hemangiomas, involuting hemangiomas, involuted hemangiomas. The mean optical density was measured by image analysis system.</p><p><b>RESULTS</b>The expression of CEACAM-1 in early stage of proliferating hemangiomas was weak or negative, while it was strong in involuting hemangiomas and positive in the involuted stage. The differences between different stages had a statistically significance (P < 0.05). The expression of CXCL-14 was weak or negative in stage of proliferating hemangiomas, positive in involuting hemangiomas and strong in the involuted stage. The differences between different stages had a statistically significance (P < 0.05).</p><p><b>CONCLUSIONS</b>CEACAM-1 and CXCL-14 are involved in the occurrence and development of infantile hemangioma.</p>

UL146 variability among clinical isolates of Human Cytomegalovirus from Japan

Human Cytomegalovirus (HCMV) is a herpesvirus associated with serious diseases in immunocompromised subjects. The region between ORF UL133 and UL151 from HCMV, named ULb' is frequently deleted in attenuated AD169 and in highly passaged laboratory strains. However, this region is conserved in low-passaged and more virulent HCMV, like the Toledo strain. The UL146 gene, which is located in the ULb' region, encodes a CXC-chemokine analogue. The diversity of UL146 gene was evaluated among fifty-six clinical isolates of HCMV from Japan. Results show that UL146 gene was successfully amplified by the polymerase chain reaction (PCR) in only 17/56 strains (30 percent), while the success rate for UL145/UL147 gene was 18/56 strains (32 percent). After DNA sequencing, the 35 amplified strains were classified into 8 groups. When compared, variability of UL146 ranged from 25.1 percent to 52.9 percent at the DNA level and from 34.5 percent to 67 percent at the amino acid level. Seven groups had the interleukin-8 (IL-8) motif ERL (Glu-Leu-Arg) CXC and one group had only the CXC motif, suggesting the absence of the IL-8 function of UL146. In conclusion, we found that UL146 gene of HCMV is hyper-variable in clinical strains from Japan suggesting the possibility of a different function in each sequence group.

Anti-mycobacterial treatment reduces high plasma levels of CXC-chemokines detected in active tuberculosis by cytometric bead array

Chemokines recruit and activate leukocytes, assisting granuloma formation. Herein, we evaluated plasma chemokines in patients with active tuberculosis (ATB) and after completing treatment (TTB) and compared them to BCG-vaccinated healthy controls (HC). Levels of chemokines were measured by cytometric bead array. Levels of CXCL8, CXCL9 and CXCL10 were higher in ATB patients compared to HC, but they decreased in TTB. Levels of CCL2 and CCL5 in ATB patients were similar to those observed in HC. Thus, the high levels of CXC-chemokines detected during ATB, which can modulate the trafficking of immune cells from the periphery to the site of infection, were reversed by anti-mycobacterial treatment.

Significance of expression of stromal cell derived factor 1 and CXCR4 in invasive breast cancer / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the expression of stromal cell derived factor 1(SDF-1)/CXCR4 and their association with clinicopathologic features and lymph node metastasis in invasive breast carcinoma.</p><p><b>METHODS</b>The expression of SDF-1 was studied by immunohistochemistry and in-situ hybridization. Immunohistochemical study for CXCR4 was also performed. The correlation with various clinicopathologic parameters was analyzed.</p><p><b>RESULTS</b>(1) SDF-1 was mainly expressed in tumor cells and the level of its expression (both membranous and cytoplasmic) in lymph node-positive group was higher than that in lymph node-negative group (P = 0.033). Only cytoplasmic expression correlated with the number of positive lymph node involved by metastasis, TNM tumor stage, histologic grade, tumor dimension and estrogen receptor status (P < 0.05). (2) SDF-1 protein was also detected in the endothelial cells, although its mRNA was rarely detected. SDF-1 staining in lymphatics was associated with positive lymph node (P = 0.005) and SDF-1 staining in blood vessels correlated with stromal lymphocytic reaction (P = 0.001). The extent of nodal involvement was higher in the group with positive SDF-1 staining in blood vessels and with prominent lymphocytic reaction than that in other groups with one or neither of the two features (P < 0.05). (3) On the other hand, CXCR4 was mainly expressed in tumor cells (both nuclear and cytoplasmic); and the level of its expression in lymph node-positive group was higher than that in lymph node-negative group (P = 0.005). Only cytoplasmic expression correlated with the number of positive lymph node involved by metastasis, TNM tumor stage, histologic grade, tumor dimension and HER2 status (P < 0.05). The nuclear expression of CXCR4 was only correlated with progesterone receptor status (P < 0.01). The cytoplasmic expression CXCR4 also positively correlated with SDF-1 expression (P = 0.001).</p><p><b>CONCLUSIONS</b>SDF-1 and CXCR4 can serve as biomarkers for diagnosis and prediction of lymph node metastasis, as well as potential therapeutic targets in invasive breast carcinoma. The difference in localization and staining patterns may also carry different significance.</p>

Chemokine SR-PSOX/CXCL16 expression in peripheral blood of patients with acute coronary syndrome / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Scavenger receptor that binds phosphatidylserine and oxidized lipoprotein/CXC chemokine ligand 16 (SR-PSOX/CXCL16) promotes foam cell formation through the tumor necrosis factor (TNF)-alpha mediated mechanism. Because chemokine CXCL16 could be expressed in atherosclerotic lesions and induce smooth muscle cell (SMC) proliferation, we presume that the monocyte SR-PSOX/CXCL16 detection in the patients' peripheral blood will be important for early diagnosis and prognosis of atherosclerosis (AS).</p><p><b>METHODS</b>Enrolled in this study were 40 patients with acute coronary syndrome (ACS), including 20 patients with acute myocardial infarction (AMI) and 20 patients with unstable angina pectoris (UAP), and 20 normal controls. Monocytes in the peripheral blood were isolated, and the changes of expression of CXCL16/SR-PSOX mRNA were compared using reverse transcription-polymerase chain reaction (RT-PCR), with beta-actin as internal control. We compared the expression of CXCL16/SR-PSOX in the ACS subgroups, using Western-blot to analyze protein expression levels. Tissue sections were made from biopsy specimens taken from patients with infective endocarditis, liver cirrhosis, and lung cancer as well as normal controls. And the expression of CXCL16/SR-PSOX was analyzed with a confocal microscope.</p><p><b>RESULTS</b>The expression of CXCL16/SR-PSOX mRNA and protein in the monocytes of peripheral blood was significantly higher in ACS patients than in normal controls (P < 0.05); however, there was no significant difference in CXCL16/SR-PSOX expression between UAP group and AMI group (P > 0.05). Immunofluorescence showed that there were low expression of SR-PSOX in normal vascular endothelial cells and enhanced expression in every layer of the infected vessels, while spreading from endothelial cells to surrounding tissues as infection worsens. Confocal microscopy showed that the expression of SR-PSOX was enhanced in the infiltrated lymphocytes in liver cirrhosis, and that the expression level was proportionate to the degree of inflammation in the portal hepatis and folia.</p><p><b>CONCLUSIONS</b>The expression of CXCL16/SR-PSOX in the monocytes of peripheral blood was significantly higher in ACS patients than in the controls. CXCL16/SR-PSOX-mediated inflammation may contribute to the pathogenesis of ACS, and CXCL16 may play an important role in the pathogenesis and development of AS in humans.</p>

Role of CXCL12 in metastasis of human ovarian cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In a previous study, we have verified that CXCR4 expression is correlated with tumor aggressive progression and poor prognosis in patients with epithelial ovarian cancer. The aim of this study was to explore the effect of CXCL12-CXCR4 axis on the metastasis of human ovarian cancer.</p><p><b>METHODS</b>The expressions of CXCR4 and CXCL12 mRNA and protein in human ovarian cancer cell line CAOV-3 was detected by RT-PCR and immunocytochemistry. Methythiazolyltetrazolium (MTT) was used to analyze the effect of different concentrations of CXCL12 on the proliferation of CAOV-3 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of CXCL12 and ascites on the migration and invasion of CAOV-3 cells. The expressions of integrin beta(1) and vascular endothelial growth factor-C (VEGF-C) mRNA were detected by RT-PCR. Data were analyzed using ANOVA by SAS 6.12.</p><p><b>RESULTS</b>Under serum-free suboptimal culture conditions, CXCL12 (100 ng/ml) significantly enhanced the proliferation of CAOV-3 cells compared with the control and 10 ng/ml CXCL12 groups (0.428 +/- 0.051 vs. 0.325 +/- 0.045 and 0.328 +/- 0.039, P < 0.05). This enhancing effect of CXCL12 was significantly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml CXCR4 antagonist AMD3100. However, 10 microg/ml neutralizing CXCR4 antibody could not inhibit cell proliferation without CXCL12. The levels of migration and invasion of the CAOV-3 cells treated with 100 ng/ml CXCL12 were significantly higher than those in the control (migration: 523.3 +/- 25.2 vs 108.0 +/- 7.2; invasion: 39.3 +/- 4.0 vs. 4.0 +/- 1.0). The enhancing effect of CXCL12 on cell migration and invasion increased with the concentration of CXCL12 (100 ng/ml vs10 ng/ml: migration, 523.3 +/- 25.2 vs 211.7 +/- 24.7; invasion, 39.3 +/- 4.0 vs 15.7 +/- 3.1, P < 0.05), and was strongly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml AMD3100. The number of migrated and invading cells in the CAOV-3 added with ascites was significantly higher than those in the 100 ng/ml CXCL12 group (migration: 706.6 +/- 30.6 vs 523.3 +/- 25.2, invasion: 61.7 +/- 7.6 vs 39.3 +/- 4.0, P < 0.05). The level of integrin beta(1) mRNA was greatly increased at 3 hours after being treated with CXCL12 (0.53 +/- 0.10 vs. 1.53 +/- 0.16, P < 0.05), and VEGF-C mRNA displayed significant augment at 24 hours after being treated with CXCL12 (0.52 +/- 0.09 vs 1.11 +/- 0.15, P < 0.05).</p><p><b>CONCLUSIONS</b>CXCL12 and its receptor CXCR4 can promote the proliferation, migration, invasion of ovarian cancer cell line CAOV-3 and enhance its secretion of integrin beta(1) and VEGF-C. These effects can be inhibited by neutralizing CXCR4 antibody or AMD3100. CXCL12-CXCR4 axis plays an important role in ovarian cancer growth and metastasis.</p>

Dexamethasone Does Not Inhibit Airway CXC Chemokine Expression and Neutrophilia in a Murine Model of Asthma - Mechanism of Steroid Resistance in Asthma

BACKGROUND: Although glucocorticoids (GCs) are effective in controlling asthma in the majority of patients, a subset of asthmatics fails to demonstrate a satisfactory response, even to systemic GC therapy. This population is referred to as being "steroid-resistant". The actual mechanism underlying steroid resistance in asthma remains to be elucidated. METHODS: We have investigated how dexamethasone (DEX) regulates asthmatic phenotypes in a murine model of asthma, in which mice received i.p. immunization twice, followed by two bronchoprovocations with aerosolized OVA with a one-week interval, which we have recently described. RESULTS: Pretreatment with DEX resulted in an inhibition of NF-kappaB activation in asthmatic lungs, and also inhibited bronchoalveolar lavage (BAL) levels of NF-kappaB-dependent cytokines such as TNF-alpha and CC chemokines [eotaxin and monocyte chemotactic protein (MCP)-1]. DEX was effective in suppressing airway hyperresponsiveness (AHR) at 10 h, Th2-dependent asthmatic phenotypes such as airway eosinophilia, BAL levels of Th2 cytokines (IL-5 and IL-13), and mucin production. However, DEX failed to suppress BAL levels of CXC chemokines [macrophage inflammatory protein-2 (MIP-2) and keratinocyte-derived chemokine (KC)] and airway neutrophilia. CONCLUSION: Airway neutrophilia is among the phenomena observed in patients with severe GC-resistant asthma. This study will provide insight into the molecular basis for airway neutrophila seen in steroid-resistant asthma. Further studies are required to delineate the underlying mechanism of CXC chemokine expression in asthma.

[Gene cloning and expression analysis of Gro/KC by primary hepatocytes]

It is now well established that several environmental stresses lead to activation of p38 MAP kinase and JNK in various cell systems which is followed by chemokine production. We investigated the expression of both CXC chemokines SDF- 1 alpha[ELR] and Gro/KC [ELR[+]] in rat H4 hepatoma cells in response to heat shock, hyper-osmolarity and oxidative stresses. The pattern of expression of these chemokines by hepatoma cells in response to stress conditions was also studied. Hepatoma cells were maintained in MEM medium. Cells were subjected to different stresses [H[2]O[2] 0.15% [w/v], manitol and NaC1 [160 mM] and heat shock [[42[degree sign] C for 20 minutes]]. At the indicated time points, cells were harvested and RNA was extracted, purified and expression of the chemokines were analysed by RT-PCR. cDNA was separated by gel electrophoresis on a 1% [w/v] agarose gel and visualized on a UV transilluminator. Results obtained in this report showed that there was detectable but low expression of chemokines in H4 hepatoma cells. Heat shock failed to induce expression of chemokines in H4 rat hepatoma cells. Hyper-osmolarity also has not stimulated Chemokines expression. In this study we have also shown that oxidative stress did not induce expression of chemokines. Overall, although detection is possible but regularly responses were not observed in H4 hepatoma cells. Several known injurious conditions cause recruitment of macrophages, neutrophils and other immune cells to the liver. Immune cells are recruited to the hepatic vasculature following local liver injury and consequent chemokine production. Our results demonstrated that failure in production of these chemokines by Hepatoma cells may be a way to escape from immune surveillance

Chemokines in the cerebrospinal fluid of patients with active and stable relapsing-remitting multiple sclerosis

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the human central nervous system. Although its etiology is unknown, the accumulation and activation of mononuclear cells in the central nervous system are crucial to its pathogenesis. Chemokines have been proposed to play a major role in the recruitment and activation of leukocytes in inflammatory sites. They are divided into subfamilies on the basis of the location of conserved cysteine residues. We determined the levels of some CC and CXC chemokines in the cerebrospinal fluid (CSF) of 23 relapsing-remitting MS patients under interferon-ß-1a therapy and 16 control subjects using ELISA. MS patients were categorized as having active or stable disease. CXCL10 was significantly increased in the CSF of active MS patients (mean ± SEM, 369.5 ± 69.3 pg/mL) when compared with controls (178.5 ± 29.1 pg/mL, P < 0.05). CSF levels of CCL2 were significantly lower in active MS (144.7 ± 14.4 pg/mL) than in controls (237.1 ± 16.4 pg/mL, P < 0.01). There was no difference in the concentration of CCL2 and CXCL10 between patients with stable MS and controls. CCL5 was not detectable in the CSF of most patients or controls. The qualitative and quantitative differences of chemokines in CSF during relapses of MS suggest that they may be useful as a marker of disease activity and of the mechanisms involved in the pathogenesis of the disease.