MCP-1/CCR2 signaling pathway mediates alcohol-induced angiogenesis in breast cancer

AIM To investigate the role of monocyte chemoattractant protein-1 (MCP-1) and its receptor CC chemokine receptor 2 (CCR2) in ethanol-promoted breast cancer angiogenesis and the underlying mechanism. METH?ODS: A mouse model of transplanted breast tumor with moderate alcohol consumption was established. The correlations between the expression of MCP-1/CCR2 and the expression of angiogenesis markers [platelet endothelial cell adhesion molecule-1 (PECAM-1) and vascular endothelial growth factor (VEGF)] in tumor tissues were examined by immunohistochemistry. In vitro, a 3D tumor-endothelial co-culture system was established to observe tumor angiogenesis and the role of MCP-1/CCR2 signaling pathway in alcohol-mediated angiogenesis. The cell migration ability was detected to clarify whether MCP-1/CCR2 enhanced cell mobility to form new vessels. RESULTS MCP-1 and CCR2 were both highly expressed in the breast tumor tissues of tumor-bearing mice consuming alcohol, and their expression levels were consistent with the angiogenic markers PECAM-1 and VEGF (P<0.05). The interaction between mouse breast cancer E0771 cells and endothelial cells was observed to promote angiogenesis in the 3D tumor-endothelial co-culture system with or without alcohol stimulation. MCP-1 promoted this kind of tumor angiogenesis, while CCR2 antagonist effectively inhibited the tumor angiogenesis and especially blocked alcohol-induced angiogenesis. Activation of MCP-1/CCR2 signaling pathway enhanced the migration ability of endothelial cells. CONCLUSION The MCP-1/CCR2 signaling pathway plays an important role in promoting the angiogenesis of breast cancer stimulated by alcohol. The mechanism might be that MCP-1 improves the migration of endothelial cells and then promotes angiogenesis.     

CCL3 promotes hBMSC proliferation and inhibits exosome secretion

AIM: To explore the regulatory effect of chemokine CCL3 on exosome secretion from human bone marrow mesenchymal stem cells (hBMSCs).METHODS: hBMSCs were stimulated with chemokine CCL3 at different concentrations in vitro. The proliferation of hBMSCs was measured by CCK-8 assay and viable cell counting. Exosome secretion from hBMSCs was qualitatively analyzed by transmission electron microscope (TEM) and flow cytometry, and the quantitative analysis was carried out by flow cytometry and nanoparticle tracking analysis (NTA).RESULTS: Compared with control group, the viability of the hBMSCs detected by CCK-8 assay was increased when hBMSCs were treated with CCL3 (P<0.05). The results of viable cell counting demonstrated that the number of hBMSCs was raised in CCL3 group in a dose-dependent manner (P<0.05). The results of flow cytometry showed that hBMSCs expressed 3 CCL3-related specific receptors, CCR1, CCR5 and CCR9. Compared with control group, the fluorescence intensity of CCR9 in CCL3 group was obviously enhanced. However, no significant difference of fluorescence intensity for CCR5 and CCR1 was observed between the 2 groups. The results of NTA demonstrated that the secretion capacity of CCL3-induced hBMSCs was far less than that in control group (P<0.05). However, the microvesicles larger than 100 nm in CCL3 groups were increased (P<0.05). The above results indicated that the higher concentration of CCL3 induced the lower secretion of exosomes. In addition, the results of flow cytometry demonstrated that CCL3-induced hBMSCs showed lower quantity of CD9⁺ exosomes than those in control group (P<0.01).CONCLUSION: CCL3 promotes the proliferation of hBMSCs but depresses the secretion of exosomes in a dose-dependent manner. CCL3 affects the size distribution of exosomes and increases the number of nonfunctional microvesicles of larger than 100 nm in size. CCL3 induces the expression of CCR9 in hBMSCs.     

Effect of Liang Xue Huo Xue capsules on mouse psoriasis-like lesions induced by imiquimod

AIM: To observe the effects of Liang Xue Huo Xue (LXHX) capsules on mouse psoriasis-like lesions induced by imiquimod (IMQ). METHODS: BALB/c female mice (n=48) were randomly divided into 6 groups: normal group, model group, LXHX capsules groups with high, medium or low doses, and glycosides of Tripterygium wilfordii (GTW) group. On day 8, skin lesions were determined by pathological examination. The lesions were evaluated according to the psoriasis area and severity index (PASI). The histology and epidermal thicknesses were observed under light microscope. The expression of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemical staining. Meanwhile, the positive expression of CD3, CD11c, F4/80, CD31 and Gr-1 was counted by immunohistochemical staining. RESULTS: Compared with model group, the cutaneous symptoms in LXHX capsules groups were alleviated, with PASI scores decreased, epidermal parakeratosis and epidermal over-proliferation relived, the numbers of dermal T lymphocytes, dendritic cells, macrophages, neutrophils and monocytes reduced significantly. CONCLUSION: LXHX capsules improve imiquimod-induced mouse psoriasis-like lesions by inhibiting over-proliferation of keratinocytes, parakeratosis, inflammatory infiltration and angiogenisis.     

Effects of Xiaoyaosan on imiquimod induced psoriatic lesions and depression neurotransmitters in mouse model

AIM: To observe the effect of Xiaoyaosan decoction on the psoriatic lesions and depression neurotransmitters induced by imiquimod in mice. METHODS: BALB/c male mice were randomly divided into control group, model group, methotrexate group and Xiaoyaosan high, medium and low dose groups, 6 mice in each group. Imiquimod (IMQ, 5%) was used on the back of the animals to induce psoriasis-like lesions in the mice. The psoriasis area and seve-rity index (PASI) were evaluated for daily scoring. The sugar water preference experiment was conducted to explore the behavioral differences in the mice. The morphological changes and epidermal thickness of the lesions were observed under light microscopy. Immunohistochemical method was used to detect the expression of CD3 on T lymphocyte surface. The expression of Ki67 in the skin lesions was detected by immunofluorescence. The contents of monoamine neurotransmitters such as adrenaline (AD), gamma-aminobutylic acid (GABA), glutamate (Glu), dopamine (DA) and their metabolites in the hippocampus and hypothalamus of mice were detected by high performance liquid chromatography-mass spectrometry (HPLC-MS). RESULTS: Compared with model group, the back skin lesions of Xiaoyaosan each dose group and methotrexate group were significantly improved, and the PASI score and epidermal thickness were both lower than those in model group (P<0.05). The expression levels of Ki67 and CD3⁺ T cells in Xiaoyaosan group and methotrexate group were lower than those in model group (P<0.05). Compared with model group, the body mass change range of Xiaoyaosan high-dose group and blank control group was significantly smaller than that in model group (P<0.05). The sugar water preference rate in blank control group was significantly higher than that in model group (P<0.01). Compared with model group, the sugar water preference rate in methotrexate and Xiaoyaosan groups showed a certain increase trend, but no statistical diffe-rence was observed. Compared model group, the levels of 3, 4-Dihydroxypheny-lacetic acid (DOPAC), AD, GLU and GABA levels in the mouse hippocampus in blank control group were decreased significantly (P<0.01), while the levels of DA and homovanillic acid (HVA) had no significant difference (P>0.05). No significant difference of DA, DOPAC, HVA and GLU levels in the mouse hypothalamus was observed between blank control group and model group (P>0.05), while the content of AD and GABA in the mouse hypothalamus in blank control group was lower than that in model group. The AD content of the hypothalamus in high-dose Xiaoyaosan group was significantly higher than that in model group (P<0.01), and the HVA content of the hypothalamus in low-dose Xiaoyaosan group was significantly higher than that in model group (P<0.01). PASI score was negatively correlated with the content of DOPAC, AD, GLU and GABA in the hippocampus and the content of AD, GLU and GABA in the hypothalamus, those were, the more severe the back skin lesion was, the lower the expression of depression-related neurotransmitters were, indicating the aggravation of depression in the mice. CONCLUSION: Xiaoyaosan improves the skin lesions induced by imiquimod in the mice with psoriasis, improves the behavior of depression in the mice with psoriasis, and up-regulates the expression of depression-related monoamine neurotransmitters. The expression of depression-related neurotransmitters is negatively correlated with the skin lesions induced by imiqumod in the mice with psoriasis. The degree of depression is increased with the aggravation of psoriatic lesions.     

Effects of Yangxue decoction and Yangxue-Jiedu decoction on imiquimod-induced psoriasis-like mouse model

AIM: To investigate and compare the effects of Yangxue (YX) decoction and Yangxue-Jiedu (YXJD) decoction on psoriasis-like mouse skin lesions. METHODS: BALB/c mice (n=50) were randomly divided into control group, model group, methotrexate (MTX) group, YX group and YXJD group (10 mice in each group). The psoriasis-like mouse model was induced by topical application of imiquimod cream on the back. The skin water/oil test pen was used to detect the water/oil content of the skin in the back of the mice. The pathological changes of the lesions were observed by HE staining and the thickness of the epidermis was measured. The immunohistochemical staining was used to observe the skin lesions, and the mRNA expression levels of interleukin (IL)-17, IL-23 and IL-1β in skin lesions were detected by real-time PCR. RESULTS: The skin lesions in YX, YXJD and MTX group were better than those in model group, with lower psoriasis area and severity index (PASI) score and skin thickness. The skin water/oil content in YXJD group was higher than that in model group (P<0.05). The expression of proliferating cell nuclear antigen (PCNA) and the positive expression of CD3⁺ T cells in the skin of YXJD group were lower than those in YX group, and the skin thickness was lower than that in YX group (P<0.05). The results of real-time PCR showed that relative mRNA expression of IL-17, IL-23 and IL-1β in YX group and YXJD group was lower than that in model group (P<0.05), and the relative mRNA expression of IL-1β in YXJD group was lower than that in YX group. Administration of YXJD decoction showed better therapeutic effect than MTX. CONCLUSION: YX decoction and YXJD decoction relieve imiquimod-induced skin lesions by reducing immune response. Meanwhile, the effect of YXJD decoction is better than that of YX decoction.     

Role of IL-23/IL-17 inflammatory axis in the pathogenesis of psoriasis-like lesions induced by imiquimod in mice

AIM: To observe the dynamic changes of IL-23/IL-17 inflammatory axis in psoriasis-like lesions of mice induced by imiquimod (IMQ).METHODS: BALB/c female mice were randomly divided into control group and IMQ group. The morphological changes of lesional skin in mice were evaluated according to the psoriasis area and severity index (PASI) and HE staining. cytokine antibody chips were used to determine the cytokine changes in serum and lesions. The mRNA and protein expression of cytokines were analyzed by cytometric bead array, real-time PCR and Western blotting. Moreover, the changes of cellular constituents in the peripheral blood and splenic cells of mice were detected by flow cytometry.RESULTS: Typical psoriasis-like skin lesions, such as red scaly skin plaques, caused by topical IMQ showed a parabolic dynamic change. There was a dynamic increase in proinflammatory cytokines of the IL-23/IL-17 axis in IMQ-treated skin. IMQ application resulted in elevated expression of cytokines related with IL-23/IL-17 inflammatory axis,Th1-type cytokines,Th2-type cytokines and Treg-type cytokines at day 4. IMQ-treated BALB/c mice showed an increased pericentage of dentric cells in peripheral blood and spleen compared with control animals. Percentages of Th17 and Treg in IMQ-treated mice were increased by 3~4 times and twice as compared with control mice, respectively.CONCLUSION: The skin lesions, histopathological features and cytokine changes in mice induced by IMQ are similar to human psoriasis, which are suitable for investigating the pathogenesis of psoriasis as a psoriasis-like model. IL-23/IL-17 axis is involved in the formation of psoriasis-like skin lesions in mice induced by IMQ and presents a dynamic change. Besides, Th1 cell-mediated inflammatory response is also activated in the formation of lesional skin, accompanied by the increase expression of Th2 and Treg cytokines in a feedback mechanism.     

Effects of marrow stromal cell-mediated microenvironment on human hepatocellular carcinoma cell line SMMC-7721

AIM: To investigate the effects of marrow stromal HS-5 cells on hepatocellular carcinoma SMMC-7721 cells in the tumor microenvironment. METHODS: The effects of HS-5 cell-conditioned medium (HS-5-CM) on the proliferation, migration and invasion abilities of SMMC-7721 cells were detected by MTT, wound-healing and Transwell assays. After co-culture of SMMC-7721 cells with HS-5 cells in the Transwell chamber, the expression of chemokine CCL5 and its receptor CCR5 at mRNA and protein levels in SMMC-7721 cells was examined by quantitative real-time PCR (qRT-PCR), ELISA or Western blotting. Akt and p-Akt473 protein levels in SMMC-7721 cells treated with PI3K inhibitor LY294002 were observed by Western blotting. RESULTS: HS-5-CM promoted the proliferation, migration and invasion abilities of SMMC-7721 cells. The expression of CCL5 and CCR5 at mRNA and protein levels in SMMC-7721 cells was increased after co-cultured with HS-5 cells. PI3K inhibitor LY294002 inhibited the activation of PI3K-Akt signaling pathway and the secretion of CCL5 in SMMC-7721 cells after co-cultured with HS-5 cells. CONCLUSION: HS-5 cells significantly promote the proliferation, migration and invasion abilities of SMMC-7721 cells. Co-culture of SMMC-7721 cells with HS-5 cells activates PI3K-Akt signaling pathway to increase the secretion of CCL5 in SMMC-7721 cells.     

Effects of Yangxue-Jiedu decoction on imiquimod-induced psoriasis-like lesions in STAT3 transgenic mice

AIM: To observe the effects of Yangxue-Jiedu (YXJD) decoction on imiquimod-induced psoriasis-like lesions in STAT3 transgenic mice.METHODS: STAT3 transgenic mice (n=24) were randomly divided into 4 groups: control group (using purified water for oral administration), model group (topical 5% imiquimod 42 mg and using purified water for oral administration), YXJD groups (topical 5% imiquimod 42 mg and using YXJD decoction for oral administration), and methotrexate (MTX) group (1 mg/kg MTX solution for oral administration, with the same topical imiquimod as model group). On day 7, the skin lesions were collected for examination. The lesions were evaluated according to the psoriasis area and severity index (PASI). The skin barrier function was evaluated by assessing oil and water components in the skin. The inflammation of psoriasis-like lesions was assessed by histological method. The expression of proliferating cell nuclear antigen (PCNA) and CD3 was assessed by immunohistochemical staining. The mRNA expression of IL-17A, IL-17C, IL-22 and RORγt was detected by real-time PCR. The levels of JAK/STAT3 pathway-related proteins in isolated T cells were determined by Western blot.RESULTS: Administration of YXJD decoction inhibited imiquimod-induced keratinocyte proliferation and infiltration of CD3⁺ T cells in psoriatic lesions, and ameliorated the epidermal barrier by up-regulation of the oil and water components in psoriatic lesions. Meanwhile, administration of YXJD decoction improved the systemic immune responses by reducing the weight of the spleen. The inflammatory cytokines IL-17A, IL-17C, IL-22 and RoRγt, and the levels of JAK/STAT3 pathway-related proteins STAT3, p-STAT3, JAK3 and p-JAK3 were decreased by administration of YXJD decoction.CONCLUSION: YXJD decoction likely alleviates imiquimod-induced psoriasis-like lesions in the STAT3 transgenic mice by inhibiting the phosphorylation of STAT3, and reducing the expression of IL-17A, IL-17C, IL-22 and RORγt.     

Change of serum CXC chemokine ligand 16 in gout patients and its clinical significance

AIM: To explore the relationship between the changes of serum CXC chemokine ligand 16 (CXCL16) and the loss of kidney functions in chronic gout patients and its clinical significance. METHODS: Twenty gout patients with chronic kidney disease (CKD), 22 gout patients without CKD and 22 CKD subjects were recruited into the present study, while 20 normal age-and sex-matched subjects were assigned into the control group. Serum level of CXCL16 and other relevant clinical and biochemical parameters in all subjects were obtained upon standard clinical examinations. Ceatinine clearance rate (CCR) and estimated glomerular filtration rate (eGFR) were calculated based on the clinical parameters. To analyze the clinical data, student's unpaired t-test was used for the comparison between 2 groups. One-way ANOVA assay and multiple stepwise regression were used for multiple groups.RESULTS: Serum level of CXCL16 was significantly increased in gout subjects compared with the healthy control and CKD subjects (P<0.05). Serum level of CXCL16 in gout patients with CKD was significantly higher than that in gout patients without CKD (P<0.05). Furthermore, the mRNA expression of CXCL16 in peripheral blood mononuclear cells (PBMC) of gout patients was significantly higher than that in healthy subjects. Multiple stepwise regression analysis indicated that serum CXCL16 was independently associated with 24 h urine protein, CCR and C-reactive protein (P<0.05). CONCLUSION: Serum CXCL16 level in gout patients is associated with the change of renal functions. Elucidating the pathophysiologcial mechanism of CXCL16 in gout patients requires further study.     

Effects of N-acetylcysteine on Clara cells and CC16 in rat COPD model

AIM: To investigate the effects of N-acetylcysteine (NAC) on the number of Clara cells and secretion of Clara cells secretory protein (CC16) in rat chrohic obstructive pulmonary disesae (COPD) model.METHODS: Thirty rats were divided into control, COPD and NAC groups (n=10). The change of Clara cell ultrastructure was detected through transmission electron microscope. The number of Clara cells and synthesis of CC16 were measured by immunohistochemistry. The CC16 levels in bronchoalveolar lavage fluid (BALF) and serum were tested by ELISA. The level of CC16 mRNA in lung was determined by RT-PCR.RESULTS: The percentage of Clara cells in terminal bronchioles in the COPD group was significantly decreased than that in the control (P<0.01), and the percentage in NAC group was significantly higher than that in COPD group (P<0.01). The levels of CC16 in the BALF and serum in COPD group were significantly lower than those in control group (P<0.01, respectively), and the levels of CC16 in NAC group were significantly higher than those in COPD group (P<0.05, respectively). The expression of CC16 mRNA in COPD group was weaker than that in control group and NAC group (P<0.01, respectively).CONCLUSION: The number of Clara cells and the secretion of CC16 decrease in a rat model of COPD. Antioxidant NAC can enhance the synthesis and secretion of CC16, which may be a mechanism for the suppression of airway inflammation.     

Effects of erythropoietin on expression of SDF-1 and its receptor in peripheral blood endothelial progenitor cells from rats with chronic renal failure

AIM:To investigate the effects of erythropoietin (EPO) on the expression of homing factors in peripheral blood endothelial progenitor cells (EPCs) from rats with chronic renal failure (CRF). METHODS:The CRF model was established by a two-stage 5/6 nephrectomy procedure in rats. Experimental rats were randomly divided into three groups: sham operation group, CRF model group and EPO treatment group. From the third week after the second stage of 5/6 nephrectomy procedure, rats in EPO treatment group received subcutaneous injection of human recombinant EPO at 50 U/kg every time and three times a week for 6 weeks, and then all the rats were sacrificed. EPCs were isolated from rat peripheral blood and primarily cultured. The mobilization, angiogenesis and functional activity of EPCs in vitro were detected. The mRNA and protein expression of EPO, EPO receptor (EPOR), stromal cell-derived factor 1 (SDF-1) and CXC chemokine receptor 4 (CXCR4) in EPCs was also detected by the methods of real-time PCR and Western blotting. RESULTS:Compared with CRF model group, the expression of EPO and EPOR in EPCs in EPO treatment group was significantly up-regulated (P<0.05). Moreover, the expression of SDF-1 and its receptor CXCR4 in EPCs was also up-regulated by administration of EPO (P<0.05). CONCLUSION: EPO can mobilize EPCs from CRF rat peripheral blood, which may be associated with the increased expression of SDF-1 and its receptor CXCR4.     

Specific surface markers of Th1/Th2 subsets and their identification

CD4+Tcells can be divided into Th1/Th2 subsets.Th1/Th2 imbalance participates many disease processes.A stable surface marker distinguishing Th1 and Th2 will greatly facilitate the investigation of Th1/Th2 interaction.Several surface molecules have been reported to be differentialy expressed between Th1 and Th2 cells.LAG-3, active ligands for P-and E-selectin, IL-18R, IL-12Rβ2, CC chemokine receptor(CCR5)were shown to be dominantly expressed on Th1 cells, whereas expression of CD30, ST2L, CRTH2, CCR3, CCR4 was reported to be preferential to Th2 cells.In this review, several surface molecules were mainly discussed.     

Biological characteristics of uNK and pNK cells in pregnancy

AIM: To study the biological characteristics of uterine natural killer cells (uNK cells) and peripheral natural killer cells (pNK cells) in early gestation and their difference on physiological functions. METHODS: MTT assay was used to detect cytotoxicity and proliferation of the uNK and pNK cells. RT-PCR technique was applied to examine the altered gene expression of the chemotaxis, angiopoiesis and other biological activity in uNK and pNK cells. RESULTS: Compared with pNK group, uNK cells showed a higher cytotoxicity against K562 and a lower proliferation activity. The expression of some chemokine receptor genes and angiogenic growth factor genes in uNK cells and pNK cells were detected, such as CCR1, CCR5, CCR7, CXCR2, CXCR4 and CX3CR1 expressed by uNK cell and high contents of CXCR4 and CX3CR1 mRNA were found. The ligand genes of the chemokine receptor were expressed by decidual tissue or trophoblast tissue in early pregnancy. PIGF and AngⅡ mRNA were only found in uNK cells. CONCLUSION: Compared with pNK cells, uNK cells have peculiarities of the behaviour that might contribute to uterine unique microenvironment during pregnancy.     

Effect of recombinant rat CC16 protein on LPS-induced expression of TNF-α, IL-6 and IL-8 in rat tracheal epithelial cells

AIM: To explore the effect of recombinamt rat CC16 protein (rCC16) on LPS-induced expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-8 in the rat tracheal epithelial (RTE) cells.METHODS: The RTE cells were incubated with rCC16 at concentrations of 0.5, 1.0 and 2.0 mg/L in serum-free media for 2 h prior to LPS (0.1 mg/L) treatment for further 24 h. The cells were harvested for assessing the mRNA levels of TNF-α, IL-6 and IL-8 by RT-qPCR. The cell culture supernatants were collected for analyzing the protein levels of TNF-α, IL-6 and IL-8 by ELISA. In addition, the nuclear translocation of nuclear factor-κB (NF-κB) p65 was tested by Western blot.RESULTS: rCC16 inhibited LPS-induced IL-6 and IL-8 expression at both mRNA and protein levels in the RTE cells in a concentration-dependent (0~2 mg/L) manner, as demonstrated by RT-qPCR and ELISA. However, no concentration-dependent manner between the dose of rCC16 and TNF-α expression was observed, and rCC16 inhibited LPS-induced TNF-α expression at lower concentration (0.5 mg/L). rCC16 concentration-dependently inhibited the effects of LPS on the level of nuclear translocation of NF-κB p65.CONCLUSION: rCC16 suppresses LPS-mediated TNF-α, IL-6 and IL-8 production through inactivation of NF-κB activity in RTE cells.[KEY WORDS] CC16 protein; Airway inflammation; LPS; Inflammatory mediators; Nuclear factor-κB     

茉莉酸甲酯诱导葡萄悬浮细胞防卫反应机制的研究

为明确茉莉酸甲酯(methyl jasmonate,Me JA)诱导葡萄果实抗病反应的机制,以‘巨峰’葡萄悬浮细胞为试材,将其继代培养一次后分别在含10μmol·L-1 Me JA的B5培养基和含100 nmol·L-1隐地蛋白的B5培养基中(模拟病原菌接种)振荡培养15 d,每3 d测定1次细胞质量及抗病相关指标。结果显示:单一Me JA处理未激活葡萄悬浮细胞内的防卫反应;而单一隐地蛋白处理则可立即显著诱导葡萄悬浮细胞内源H2O2迸发,提升vv NPR1.1、PR1和PR2表达水平和植保素含量;经Me JA处理的葡萄悬浮细胞在添加隐地蛋白后,其细胞内出现较单一隐地蛋白处理更为显著的H2O2迸发、PR基因表达和植保素合成现象,说明经Me JA处理的悬浮细胞只在病原激发子胁迫时才表达出较强的系统抗性。因此,Me JA诱导的葡萄悬浮细胞抗病反应可归因于Priming(植物敏化过程或防御准备过程)机制。此外,经Me JA诱导的Priming反应对葡萄悬浮细胞生长无显著影响,暗示其未抑制葡萄细胞生长和胞内物质积累。     

Expression of chemokine receptor CCR7 and VEGF-C is associated with prognosis of breast carcinoma

AIM: To explore the protein levels of chemokine receptor 7 (CCR7) and vascular endothelial growth factor (VEGF)-C in breast carcinoma, and to investigate the effects of CCR7 and VEGF-C on prognosis of breast carcinoma. METHODS: The protein expression levels of CCR7 and VEGF-C in the breast carcinoma tissues and normal breast tissues were detected by the method of immunohistochemistry. At the same time, the relationship between clinicopathologic characteristics and the protein expression of CCR7 and VEGF-C in the breast carcinoma tissues was analyzed. The relationship between the protein expression of CCR7 and VEGF-C and survival time of the breast cancer patients was estimated by Kaplan-Meier method.RESULTS: The positive expression rates of CCR7 and VEGF-C in the breast carcinoma tissues were significantly higher than those in the normal breast tissues (P<0.01). A positive correlation was observed between the protein expression of CCR7 and the protein expression of VEGF-C in the breast carcinoma tissues (r=0.613, P<0.01). The protein expression of CCR7 and VEGF-C was correlated with lymph node metastasis and TNM stage (P<0.05), but both were not related to patients' age, primary tumor size, estrogen receptor and progesterone receptor. The survival time of the patients with CCR7 and VEGF-C positive expression was significantly shorter than that of the patients without the expression (P<0.05).CONCLUSION: The positive expression of CCR7 and VEGF-C proteins is associated with the prognosis of breast cancer, and combined detection of CCR7 and VEGF-C protein expression levels may be helpful to judge the prognosis of breast cancer.     

Roles of atypical chemokine receptors in progression and metastasis of tumor

Chemokines and their receptors have been implicated mostly in tumor progression and metastasis. Atypical chemokine receptors (ACKRs) comprise a group of 7-transmembrane domain proteins structurally similar to G protein-coupled receptors. However, ACKRs do not induce classical signaling via the typical G protein-mediated pathways. ACKRs efficiently internalize the cognate chemokine ligands and act as scavengers instead. ACJRs are composed of at least 3 members of chemokine receptors:Duffy antigen receptor for chemokines (DARC, also known as ACKR1), D6 (also known as ACKR2) and ChemoCentryx chemokine receptor (CCX-CKR, also known as ACKR4). These receptors bind to and/or internalize their chemoattractant ligands without activating signal transduction cascades leading to cell migration. In this review, we summarize the recent progress regarding the roles of ACKRs in the progression and metastasis of tumor.     

Molecular mechanism of BMSC intracerebral transplantation in improving learning and memory abilities of AD mice

AIM: To investigate the effect of bone marrow mesenchymal stem cell (BMSC) transplantation on learning and memory abilities and pathological changes of Alzheimer disease (AD) mice and the molecular mechanisms. METHODS: C57/BL6 wild-type (WT) and transgenic (Tg) mice were randomly divided into 4 groups:WT/PBS group, WT/BMSCs group, Tg/PBS group and Tg/BMSCs group. The mice were administered with PBS or BMSCs via intracerebroventricular injection. Spatial learning and memory abilities of the mice were evaluated by Morris water maze test on the 3rd day after surgery. Real-time PCR was applied to detect the mRNA expression of CX3C chemokine ligand 1 (CX3CL1), CX3C chemokine receptor 1 (CX3CR1), IL-1β, TNF-α, Nurr1, YM1, insulin-degrading enzyme (IDE) and matrix metalloproteinase 9 (MMP9). The protein levels of CX3CL1 and Aβ42 were measured by ELISA. Western blot was used to detect the protein expression of postsynaptic density protein 95 (PSD95) and synaptophysin (SYP). RESULTS: The transplanted BMSCs were observed near the hippocampus of APP/PS1 mice on the 10th postoperative day. The escape latency of the mice in Tg/PBS group was significantly longer than that in the WT/PBS mice (P<0.05). Compared with Tg/PBS group, the escape latency of Tg/BMSCs group was significantly shorter (P<0.05), and the mRNA and protein levels of CX3CL1 in Tg/BMSCs group were significantly higher than those in Tg/PBS group (P<0.01). The results of immunohistofluorescence staining showed that BMSC transplantation promoted the activation of microglia in the brain of WT and Tg mice. The mRNA expression of YM1 was up-regulated in WT/BMSCs group and Tg/BMSCs group (P<0.05). Compared with WT/PBS mice, the mRNA expression of TNF-α in the cortex and hippocampus of Tg/PBS group was significantly increased (P<0.05), and the mRNA expression of Nurr1 in the cortex was significantly decreased (P<0.01). Meanwhile, the mRNA expression of TNF-α in the cortex of Tg/BMSCs mice was decreased (P<0.01) and the mRNA expression of CX3CR1 and Nurr1 was up-regulated compared with Tg/PBS group (P<0.05). The results of Western blot showed that the protein levels of PSD95, p85, p110 and p-Akt in Tg/BMSCs group were significantly higher than those in Tg/PBS group (P<0.05). Finally, BMSC transplantation reduced the protein level of Aβ42 in APP/PS1 mice (P<0.05), and increased the mRNA expression of IDE and MMP9 in the hippocampus (P<0.05). CONCLUSION: BMSC transplantation modulates neuroinflammatory responses and promotes neuroprotective factor and synaptic protein expression, thus improving the learning and memory abilities in the APP/PS1 mice, which may be achieved by up-regulating the expression of CX3CL1.     

Expression of CXC chemokine receptor 7 in atherosclerotic ApoE-deficient mice and therapeutic impact by atorvastatin

AIM: To evaluate the expression level of CXC chemokine receptor 7 (CXCR7) in atherosclerotic apolipoprotein E-deficient (ApoE^-/-) mice induced by high-fat diet (HFD) and the effects of atorvastatin on it. METHODS: ApoE^-/- male mice (8-week-old) were used and were randomly divided into 3 groups following 1-week normal rodent diet: normal diet control (NDC) group, HFD group and HFD+statins (HFD+Sat) group. HE staining and oil red O staining were used to observe the atherosclerotic lesion burdens in the aortas. The expression of CXCR7 on the aortas was detected by Western blot and immunohistochemistry. The expression of Akt and endothelial nitric oxide synthase (eNOS) in the aorta was determined by Western blot.RESULTS: Few lesions were found in the aortas in NDC group. Apparent atherosclerotic plaque burdens were seen in HFD group and HFD+Sat group, while the atherosclerotic plaque burdens in HFD+Sat group were notably reduced compared with HFD group. The protein levels of CXCR7, eNOS and Akt in aorta in HFD group and HFD+Sat group were significantly decreased compared with NDC group, while those in HFD+Sat group were increased compared with HFD group. The protein level of p-eNOS in the aorta and the concentration of NO in the plasma in HFD group were decreased compared with NDC group and HFD+Sat group. CONCLUSION: In ApoE^-/- mice, HFD increases the lipid level and promotes the development of atherosclerosis by downregulating the expression of CXCR7, Akt and eNOS. Atorvastatin reverses the above effect of hypercholesterolemia on the expression of CXCR7, Akt and eNOS, thus playing the role in treating atherosclerosis.