Relationship between serum biomarker levels and severity of coronary atherosclerosis / 心血管康复医学杂志

Objective :To explore relationship between serum biomarker levels and severity of coronary atherosclerosis (CAS) ,and provide reference for diagnosis and treatment .Methods : A total of 160 CAS patients treated in our hos-pital from Apr 2012 to Nov 2016 were selected .According to plaque characteristics ,patients were divided into stable plaque group (n＝80) and unstable plaque group (n＝80) ,another 60 healthy subjects undergoing physical examina-tion simultaneously were selected as healthy control group .Serum biomarker levels were measured and compared a-mong three groups ,and their relationship with CAS severity were analyzed .Results : Along with CAS aggravated , there were gradual rise in serum levels of trasforming growth factor (TGF)-β ,tumor necrosis factor (TNF)-α ,nu-clear factor (NF)-κB ,vascular endothelial growth factor (VEGF) ,recombinant bovine basic fibroblast growth fac-tor (bFGF) ,monocyte chemoattractant protein (MCP)-1 ,stroma-cell derivated factor (SDF)-1 ,chemokine re-ceptor (CXCR )-4 ,chemokine receptor (CCR )-2 ,Smad-1 ,Smad-2 ,Smad-3 ,peroxisome proliferators-activated receptor (PPAR)-γ ,tissue inhibitor of metalloproteinase (TIMP)-1 ,TIMP-2 ,TIMP-3 ,healthy control group＜stable plaque group＜ unstable plaque group ,there existed significant difference between any two groups , P＝0-001 all.Compared with healthy control group ,there were significant rise in serum levels of Caspase-3 ,Caspase-6 and Bcl-2 related X protein (Bax) ,and significant reduction in Bcl2 level in stable and unstable plaque group , P＝0-001 all.Conclusion : Biomarkers ,such as TGF-β ,MCP-1 ,CXCR-4 etc .,are closely associated with severity of coronary atherosclerosis ,which can be used as important indexes for clinical treatment .

Expression and function of chemokine TARC/CCR4 at fetal-maternal interface in first trimester / 中华妇产科杂志

Objective To investigate the expression and function of thymus and activation regulated chemokine (TARC) and its special receptor CCR4 at placenta villous in the first trimester placenta villous.Methods Placenta villous was collected from healthy women undergoing artificial abortion at 6 to 8 weeks of gestation,mRNA levels of TARC,CCR4 were analyzed using semi-quantitative reverse transcription (RT)-PCR methods.Immunohistochemistry assay was used to assess the protein localization and expression of TARC,CCR4.Additionally,extravillous cytotrophoblasts were isolated and cultured.Expression of TARC and CCR4 was measured by immunofluorescence assay.Invasion of cell line HTR8/SVneo was analyzed by transwell assay at concentration of 10,25,50 and 100 ng/ml of TARC matched with RPMI 1640 fetal bovine serum free eulture medium as control group.In the mean time,blocking experiment was also added to detect TARC regulating cell invasion,which were classified into four groups:control,100 ng/ml rhTARC,20 μg/ml anti-TARC + 100 ng/ml rhTARC,100 ng/ml rhTARC + 20 μg/ml IgG.The influence of 100 ng/ml TARC on expression level of integrin-α5 and integrin-β1 were measured by using western-blot assay.Results (1)In vivo assay:expression of TARC and CCR4 mRNA were detectable in first trimester placenta villous,TARC protein was localized in cytotrophoblasts,syncytiotrophoblasts and cell column especially on the distal portion,while CCR4 protein was localized on invading interstitial cytotrophobalsts.(2)In vitro assay:①TARC,CCR4 was also expressed in primary isolated extravillous cytotrophoblasts by immunofluorescence assay; ②Matrigel invasion assay demonstrated that TARC had specific dose dependent stimulatory effects on the cells invading through the matrigel precoated filter,the number of cells migration into the lower chamber were:142 ± 31 at 10 ng/ml group,161 ±46 at 25 ng/ml group,201 ±30 at 50 ng/ml group,312 ±48 at 100 ng/ml group,117 ± 33 at control group,the significant response observed from 25 ng/ml (P ＜ 0.05)and reached a peak effect at 100 ng/ml (P ＜ 0.01); ③Blocking experiment demonstrated that when trophoblast invasion was monitored in response to TARC neutralizing antibody (15 μg/ml) together with rhTARC 100 ng/ml.The stimulatory activity of rhTARC was completely overcome,with the cells invasion into the lower chambers were 100 ng/ml rhTARC,20 μg/ml anti-TARC + 100 ng/ml rhTARC,100 ng/ml rhTARC +20 μg/ml IgG,control:313 ±47,113 ±41,287 ±75 and 128 ±23,respectively;④Western-blot assay demonstrated that if cells were treated with 100 ng/ml rhTARC,the expression of integrin-α5 were significantly increased(P ＜0.01),integrin-β1 level also increased when compared with control(P ＜0.05).Conclusion TARC was expressed specifically at human fetal-maternal interface.Trophoblast invasion and migration mainly was regulated by up-regulation integrin-α5 and integrin-β1,which plays an role in trophoblasts differentiation and placentation.

Expression and clinical significance of CXCR4, CXCL12 and PTEN in breast cancer / 国际肿瘤学杂志

Objective To analyse the expressions and clinical significances of CXCR4,CXCL12 and PTEN in breast cancer.Methods The expressions of CXCR4,CXCL12 and PTEN in 60 cases of breast cancer were detected by immunohistochemistry technique SABC method.The correlations of levels of CXCR4,CXCL12 and PTEN expression and the age of patient,tumor diameter,histological grade,TNM stage,lymph node metastasis,ER status,Her-2 status,and vessel invasion were analysed.Twenty cases of breast fibroadenoma tissues and 20 cases of normal breast tissues were analysed as controls.Results The expressions of CXCR4 (x2 =48.754,P =0.000),CXCL12 (x2 =47.611,P =0.000) and PTEN (x2 =19.994,P =0.000) in breast cancer,normal breast tissues and breast fobroadenoma showed significant differences.The positive expressions of CXCR and CXCL12 were significantly correlated with histological grade (x2 =11.080,P =0.004;x2 =6.978,P =0.031),TNM stage (x2 =9.819,P =0.007;X2 =10.163,P =0.006),lymph node metastasis (x2 =6.213,P =0.013;x2 =8.031,P =0.005),ER (x2 =12.774,P =0.000;x2 =7.330,P=0.007),vessel invasion (x2 =5.860,P=0.013; x2 =5.185,P=0.020) and Her-2 (x2 =5.487,P =0.019;x2 =4.689,P =0.030).The expression of PTEN in breast cancer was significantly correlated with TNM stage (x2=7.366,P =0.025),lymph node metastasis (x2 =5.511,P =0.019) and ER state (x2 =4.077,P =0.043).There was a positive correlation between the expression of CXCR4 and the expression of CXCL12 in breast cancer (r =0.336,P =0.004).There was a negative correlation between the expression of CXCR4 and the expression of PTEN in breast cancer (r =-0.362,P =0.004).There was a negative correlation between the expression of CXCL12 and the expression of PTEN in breast cancer (r =-0.360,P =0.004).Conclusion The high expression of the chemokine CXCL12 and its receptor CXCR4 and the low expression of PTEN are closely related to the carcinogenesis and metastasis of breast cancer,which may play an important role in the development of breast cancer.

Effect of the monocyte locomotion inhibitory factor (MLIF) produced by E. histolityca on cytokines and chemokine receptors in T CD4+ lymphocytes

Entamoeba histolytica produces Monocyte Locomotion Inhibitory Factor (MLIF), which may contribute to the delayed inflammation observed in amoebic hepatic abscesses. Leukocytes are affected through the modulation of cytokine expression and/or production. We evaluated the effects of MLIF on the activation and production of intracellular cytokines in human CD4+ T lymphocytes by flow cytometry. Cells were stimulated for 24 h with PMA, MLIF, or PMA+MLIF. Cellular activation was measured using anti-CD69. Th1/Th2 production was studied by the expression of intracellular cytokines and cytokine/chemokine receptors. MLIF increased CD69 and induced the over-expression of the IL-l©¬, IFN-¥ã, IL-2, IL-4, and IL-10 intracellular cytokines; PMA+MLIF inhibited Th1 cytokine (IFN-¥ã) and increased Th2 cytokines (IL-4 and IL-10). The co-expression of the cytokine and chemokine receptors IFN-¥ã/CCR5 and IL-1©¬/CCR5 was inhibited by PMA+MLIF and Th2 co-expression was increased. MLIF effects varied depending on the conditions. MLIF alone activated the Th1 and Th2 cytokines and cytokine/receptor expression; however, PMA+MLIF increased the expression of Th2 but inhibited it in Th1.

Selective addition of CXCR3+CCR4-CD4+ Th1 cells enhances generation of cytotoxic T cells by dendritic cells in vitro

Increasing importance is being given to the stimulation of Th1 response in cancer immunotherapy because its presence can shift the direction of adaptive immune responses toward protective immunity. Based on chemokine receptor expression, CXCR3+CCR4-CD4+ T cells as Th1-type cells were investigated its capacity in monocyte-derived dendritic cell (DC) maturation and polarization, and induction of antigen specific cytotoxic T lymphocytes (CTL) in vitro. The levels of IL-4, IL-5 and IL-10 were decreased to the basal level compared with high production of IFN-gamma, TNF-alpha, and IL-2 in CXCR3+CCR4-CD4+ T cells stimulated with anti-CD3 and anti-CD28 antibodies. Co-incubation of activated CD4+ or CXCR3+CCR4-CD4+ T cells with DC (CD4+/DC or CXCR3+CD4+/DC, respectively) particularly up-regulated IL-12 and CD80 expression compared with DC matured with TNF-alpha and LPS (mDC). Although there was no significant difference between the effects of the CXCR3+CCR4-CD4+ and CD4+ T cells on DC phenotype expression, CXCR3+CD4+/DC in CTL culture were able to expand number of CD8+ T cells and increased frequencies of IFN-gamma secreting cells and overall cytolytic activity against tumor antigen WT-1. These results demonstrated that the selective addition of CXCR3+CCR4-CD4+ T cells to CTL cultures could enhance the induction of CTLs by DC in vitro, and implicated on a novel strategy for adoptive T cell therapy.

Roles of the chemokine CCL22 and its receptor CCR4 in the pathogenesis of systemic lupus erythematosus / 中华皮肤科杂志

Objective To investigate the expression of cellular chemokine CCL22 and its receptor CCR4, as well as its clinical significance in systemic lupus erythematosus (SLE), along with its roles in the pathogenesis of this disease. Methods Forty-eight patients with SLE and 26 normal human controls were recruited into this study. The patient cohort included 2 males and 46 females with an average age of 33.98± 12.73 years and disease course of 1 month to 20 years. Blood samples were collected from the subjects. ELISA and flow cytometry were used to examine the plasma concentration of CCL22 together with the CCR4 expression on peripheral blood cells. SLEDAI was applied to evaluate the severity of SLE patients. Results The plasma concentration of CCL22 was 227.03±122.84 ng/L in SLE group, 369.53±79.10 ng/L in the control group, 168.09±61.83 ng/L in patients with lupus nephritis and 292.77±163.45 ng/L in patients without lupus nephritis; there was a significant difference between the SLE patients and normal con-trols (P < 0.05) as well as between patients with lupus nephritis and those without (P < 0.05). Increased percentage of CCR4-expressing cells were observed in the peripheral blood of patients with SLE compared with the controls (20.24%±13.86% vs 10.44%±3.07%, P < 0.01), and the percentage of CD3+CCR4+ cells was significantly higher than that of CD3-CCR4+ cells. Moreover, a decrease was noted in the plasma con-centration of CCL22 in severe patients (P < 0.05). In SLE patients, the percentage of CCR4 increased with the rise in SLEDAI score, whereas the plasma concentration of CCL22 negatively correlated with SLEDAI score (r = -0.308, P < 0.05). Conclusion CCL22/CCR4 may play a certain role in the development, pro-gression and organ involvement in SLE.