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AIM:To investigate the roles of Notch signaling in lipopolysaccharide(LPS)-induced proliferation and secretion of interleukin-6(IL-6)and chemokine CXCL1 in bone marrow mesenchymal stem cells(BMSCs). METHODS:BMSCs were isolated by whole bone marrow culture.The expression levels of Notch signaling pathway recep-tors and ligands in the BMSCs treated with LPS were measured by qPCR and Western blot.The proliferation of BMSCs was analyzed by MTT assay and viable cell counting.The secretion levels of IL-6 and CXCL1 induced by LPS were measured by ELISA.RESULTS:Treatment with LPS at 1 mg/L effectively induced the proliferation of BMSCs and the secretion of IL-6.Obvious expression of Notch receptors and ligands in the BMSCs was observed,and LPS had little effect on the mRNA and protein levels of Notch receptors and ligands,but LPS increased the protein levels of Hes1 and Hey1,the target genes of Notch signaling.LPS at 1 mg/L increased the proliferation of BMSCs,whereas DAPT(Notch signal inhibitor)reduced the basal and LPS-induced proliferation of BMSCs(P<0.01).LPS treatment robustly increased the secretion of IL-6 and CXCL1 as assessed by ELISA.However,inhibition of Notch signaling almost completely abolished LPS-induced secretion of IL-6 and CXCL1(P <0.05).CONCLUSION: Inhibition of Notch signaling reduced not only the proliferation of BMSCs but also IL-6 and CXCL1 secretion induced by LPS.
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Objective To explore the relationship between CXCL1 expression level and prognosis of HBV-associated hepatocellular carcinoma (HCC) after liver transplantation.Methods From 2000 to 2010,127 recipients of liver transplantation for HBV-associated HCC in our hospital were enrolled.General hematoxylin-eosin staining and CXCL1 immunohistochemical staining were applied in formalin-fixed paraffinembedded sections,and the Cohort's adverse prognostic factors were analyzed retrospectively.Neutrophil migration assay and Transwell invasion assay were used to verify its mechanism.Results CXCL1 expression level is an independent prognostic factor for 5-year overall survival (OS) and disease-free survival (DFS) (P ＜ 0.05);Neutrophil migration assay and Transwell invasion assay suggested that HCC cells can secrete CXCL1 recruiting neutrophils to hepatic carcinoma tissues,and neutrophils secrete vascular endothelial growth factor A (VEGFA) promoting the invasion of HCC cells through vascular endothelial growth factor receptor 2 (VRGFR2).Conclusion High CXCL1 level in HCC tissues is an independent prognostic factor for HCC recurrence and long-term survival of patients with HBV-associated HCC after liver transplantation.CXCL1-neutrophil-VEGFA-VEGFR2 may be a mechanisms leading to HCC recurrence in patients with HBV-associated HCC after liver transplantation.
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Objective To investigates the diagnostic value of combined detection serum CCL18, CXCL1 antigen, C1D, TM4SF1, FXR1, TIZ IgG autoantibody by suspension array for ovarian cancer. Methods Suspension array was used to detect CCL18, CXCL1 antigen, C1D, TM4SF1, FXR1, TIZ IgG autoantibody in 120 cases of healthy women, 204 cases of patients with benign pelvic tumors, 119 cases of pelvic malignant tumor patients, and 40 cases with breast cancer, lung cancer oroliver cancer, respectively. Constructed diagnosis model of combined detection six biomarkers for diagnosis of ovarian malignant tumor. Constructed diagnosis model of combined detection autoantibodies to diagnose epithelial ovarian cancer. Analysed the value of detecting six biomarkers for diagnosis of ovarian malignant tumor and detecting autoantibodies for diagnosis of epithelial ovarian cancer. Analysed diagnostic value of detecting six biomarkers to diagnose stageⅠandⅡepithelial ovarian cancer. Compared diagnostic value of detecting six biomarkers in diagnosis of tissue types and pathologic grading with that of CA125. Results Model of combined detecting six biomarkers to diagnose ovarian malignant tumor was logit(P)=-11.151+0.008×C1D+0.011 × TM4SF1+0.011 × TIZ-0.008 × FXR1+0.021 × CCL18+0.200 × CXCL1. Model of combined detection autoantibodies to diagnose epithelial ovarian cancer was logit(P)=-5.137+0.013 × C1D+0.014 × TM4SF1+0.060 × TIZ-0.060 × FXR1. Sensitivity and specificity of detecting six biomarker to diagnose ovarian malignant tumor was 90.6% and 98.7%. Sensitivity and specificity of detecting autoantibodies to diagnose epithelial ovarian cancer was 75.8%and 96.7%. Combined detection for six biomarkers to diagnose serous and mucinous ovarian cancer was statistically no better than those of CA125 (P=0.196 and P=0.602, respectively);there was significantly difference in diagnosis of ovarian cancer (P=0.023), and there was no significantly difference in diagnosis of different pathological grading (P=0.089 and P=0.169, respectively). Conclusions Constructing diagnosis model of combined detection six biomarker to diagnose ovarian malignant tumor and constructed diagnosis model of combined detectionautoantibodies to diagnose epithelial ovarian cancer. Combined detection six biomarkers to diagnose serous and mucinous ovarian tumors is better than that of CA125.
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Objective To investigate the chemokine 12 (CXCL12) and chemokine receptor 4 (CXCR4) expressions in hypopharyngeal carcinoma and its place in the disease development,invasion and metastasis of significance.Methods Immunohistochemistry was used to detect the expressions of CXCL12 and CXCR4 in 35 cases of hypopharyngeal cancer tissues and in 28 cases of tumor-adjacent non-tumor tissues.Results The expressions of CXCL12 and CXCR4 in the hypopharynx carcinomas were significantly higher (P ＜ 0.05).Both expressed in hypopharyngeal carcinomas was significantly positively correlated (P ＜ 0.01).Both hypopharynx cancer in lymph node metastasis group were significantly higher than the expression of cervical lymph node metastasis group,the difference was significant (P ＜ 0.05).Conclusions CXCL12 and CXCR4 are involved in hypopharynx cancer development,invasion and metastasis,and there is a positive feedback regulation mechanism between two factors.Moreover,CXCL12 and CXCR4 have synergistic effect in development,invasion and metastasis of hypopharynx cancers.
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Objective To observe the effect of intrathecal injection (IT) of oxycodone hydrochloride on neuropathic pain and spinal cord level of microglial c-Jun N-terminal kinase/chemokine (C-X-C motif) ligand 1 (c-JNK/CXCL) 1signal in rat model of chronic constriction injury (CCI).Methods Male Sprague-Dawley rats were randomly divided into five groups (n =40 per group):sham group (intrathecal normal saline,IT NS),CCI group (CCI + IT NS),oxy group (CCI + IT 5 μg/30 μl oxy),mino group (CCI + IT 5 μg/30 μl Minocycline),and c-JNK inhibitor group (SP group,CCI + IT 5 μg/30 μl SP600125).The lumbar intrathecal catheters were implanted in L5-6 of rats and CCI models were established as previously described.The thermal and mechanical nociceptive thresholds were assessed by paw withdrawal latency (PWL) to radiant heat and von Frey filaments.The oxycodone,minocycline and SP600125 were administered intrathecally for 3 days before surgery.The spinal cord expression of Ⅰ ba-1,p-c-JNK and CXCL1 proteins assessed by Western blot.Immunofluorescence staining was performed to examine microglia morphology and the number of Ⅰ ba-1 positives cells in dorsal horn of injured spinal cord at 7 days post-IR.Results Compared to sham group,rats in CCI group had significantly lower mechanical and thermal pain thresholds,but higher spinal proteins expression of Ⅰ ba-1,and p-c-JNK and CXCL1 (P ＜0.05).Rats in oxy group,mino group and SP group had significantly higher mechanical and thermal pain thresholds and significantly lower proteins expression of Ⅰ ba-1,p-c-JNK and CXCL1 compared to those in CCI group (at any observed time-point after ligation,but most significantly at 7days,P ＜ 0.05).At the 7days after surgery,microglial cells in CCI group transformed from the ramified shape to amoeboid macrophage-like shape by immunofluorescence staining with the increases of Ⅰ ba-1 positive cells;while the other three groups exhibited hypertrophic morphology with less number Ⅰ ba-1 positive cells (P ＜ 0.05).There were no significant differences between these three groups at any observed time (P ＞ 0.05).Conclusions Intrathecal injection of oxycodone hydrochloride can relieve CCI-induced neuropathic pain by down-regulation microglial c-JNK/CXCL1 signal in spinal cords.Provide new therapeutic targets for clinical treatment of neuropathic pain.
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Objective:To compare the effects of stromal cell-derived factor-1 (SDF-1 )and granulocyte colony-stimulating factor (G-CSF)on proliferation,migration,and odontoblastic differentiation of human dental pulp stem cell (DPSC)in vitro.Methods:DPSCs were cultured in vitro and treated with either 1 00 μg/L SDF-1 or 1 00 μg/L G-CSF.Cell counting kit-8 (CCK-8 )and colony-forming unit (CFU ) were used to detect the effect of SDF-1 and G-CSF on the proliferation ability of DPSC.Cell migration of DPSC was determined by wound healing assay and Transwell migration assay.The effects of SDF-1 and G-CSF on odontoblastic differentiation of DPSC were evaluated by alkaline phosphatase (ALP)staining, ALP activity and alizarin red S staining.The expression of odontoblastic-related genes such as dentin ma-trix protein 1 (DMP-1 )and dentin sialophosphoprotein (DSPP)were quantified by real-time RT-PCR. Results:SDF-1 and G-CSF promoted the proliferation of DPSC slightly,but the difference was not statis-tically significant.Wound healing assay showed that SDF-1 and G-CSF promoted cell migration of DPSC significantly (P<0.01 ),but there was no significant difference between the two factors.In Transwell migration assay,the number of migrated cells of the control group was 5 .0 ±1 .4 per sight,while the SDF-1 group was 24.3 ±6.8 per sight and the G-CSF group was 1 1 .8 ±3.3 per sight,suggesting that cell migration of DPSC was improved significantly after being treated with SDF-1 or G-CSF,and SDF-1 was more effective than G-CSF (P<0.05 ).Significantly greater odontoblastic differentiation potential was found in SDF-1 group and G-CSF group based on the ALP staining.Higher ALP activity,more mineralization nodule formation and higher expressions of DMP-1 and DSPP were also found after SDF-1 or G-CSF treatment.Conclusion:SDF-1 had no significant effect on the proliferation of DPSC,but could significantly promote cell migration and odontoblastic differentiation of DPSC.Its effect on DPSC was bet-ter than G-CSF.
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Objective To explore the changes of GRO-α and VEGF in the peritoneal fluid of women with endometriosis and their correlation. Methods The levels of GRO-α and VEGF were detected by enzyme linked immunosorbent assay ( ELISA ) method in 39 cases of endometriosis and 25 normal cases of without endometriosis. Results The levels of GRO-αand VEGF in the peritoneal fluid of EMs group were ( 108.77 ± 20. 58 ) pg/ml and ( 199. 67 ± 99. 26 ) pg/ml separately, and it was significantly higher than those in control group ( 71.07 ± 18. 96 ) pg/ml, ( 115.46 ± 32. 39 ) pg/ml. The differences were statically significant( P <0. 05). The level of GRO-α in the peritoneal fluid of the patients in stage Ⅰ and Ⅱ was sig-nificantly lower than that in stage Ⅲ and Ⅳ[(99.43 ± 16.05)pg/ml vs (117.65 ± 20.8)pg/ml, P <0. 05] , while the level of VEGF in the peritoneal fluid of the patients in stage Ⅰ and Ⅱ was significantly higher than that in stage Ⅲ and Ⅳ[(246. 89 ± 114. 34) pg/ml vs ( 152. 82 ± 54. 55 ) pg/ml, P < 0. 05] .There was positive correlation between the level of GRO-α and r-AFS ( r = 0. 439, P < 0. 05 ), but no cor-relation between the level of VEGF and r-AFS ( r = -0. 210, P >0.05). There was no correlation be-tween peritoneal fluid levels of GRO-α and VEGF in the EMs group ( r =-0. 05, P>0. 05). Conclusion Compared with control group, the levels of GRO-α and VEGF in peritoneal fluid of women with endome-triosis were significantly increased, which indicated that the increased GR0-α and VEGF secretion involved in the pathogenesis of this disease. There was correlation between the peritoneal fluid level of GRO-α and the degree of EMs.