Therapeutic Effect of Ziziphi Spinosae Semen-Albiziae Flos on Depression in Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Ziziphi Spinosae Semen-Albiziae Flos on the nucleotide-binding oligomerization domain，NOD-like receptor thermal protein domain associated protein 1 （NLRP1）/chemokine ligand 1 （CXCL1）/chemokine receptor 2 （CXCR2） pathway in the hippocampus of the rat model of chronic unpredictable mild stress （CUMS）-induced depression. MethodA total of 120 male SD rats were randomized into blank，CUMS，CUMS + low-，medium-，and high-dose （4，8，16 g·kg-1） Ziziphi Spinosae Semen-Albiziae Flos，and CUMS + venlafaxine hydrochloride （0.008 g·kg-1） groups，with 20 rats in each group.The rat model of depression was established by solitary feeding combined with CUMS.The behaviors and spatial learning and memory abilities of rats were examined by sugar water consumption test，tail suspension test，forced swimming test，and Morris water maze test.Quantitative real-time PCR （Real-time PCR） and Western blot were employed to determine the expression of factors associated with the NLRP1/CXCL1/CXCR2 pathway in the hippocampus.Enzyme-linked immunosorbent assay was employed to determine the levels of tumor necrosis factor-α （TNF-α），interleukin （IL）-18，IL-1β，and IL-6 in the hippocampus.The immunofluorescence assay was used to measure the levels of reactive oxygen species （ROS） in the hippocampus. ResultCompared with the blank group，the CUMS group showed decreased preference to sugar water and times of crossing the platform （P<0.01），and increased immobility time of tail suspension，forced swimming floating time，and escape latency （P<0.01）.Compared with the CUMS group，the administration of Ziziphi Spinosae Semen-Albiziae Flos and venlafaxine hydrochloride alleviated the effects of CUMS on the above-mentioned behaviors and spatial learning and memory abilities of the rats （P<0.05，P<0.01）.Compared with the blank group，the CUMS group showed up-regulated protein levels of NLRP1，CXCL1，and CXCR2 （P<0.01） and elevated levels of IL-18，IL-1β，TNF-α，and IL-6 （P<0.01） in the hippocampus.The treatment with Ziziphi Spinosae Semen-Albiziae Flos and venlafaxine hydrochloride attenuated the activation of NLRP1/CXCL1/CXCR2 signaling pathway and lowered the levels of inflammatory cytokines in the hippocampus of CUMS rats （P<0.05，P<0.01）.In addition，Ziziphi Spinosae Semen-Albiziae Flos lowered the level of ROS in the hippocampus （P<0.05，P<0.01）. ConclusionZiziphi Spinosae Semen-Albiziae Flos can mitigate the depressive behaviors of the rat model of CUMS-induced depression by inhibiting the activation of NLRP1/CXCL1/CXCR2 signaling pathway.

Research progress on the role of CXCL1 in tumorigenesis and development / 中国肿瘤生物治疗杂志

@#[摘 要] 趋化因子（C-X-C基序）配体1（CXCL1）是CXC趋化因子家族中的一员，在炎症形成、新血管生成和肿瘤形成中都具有重要功能。CXCL1可通过自分泌途径促进正常成纤维细胞转化为肿瘤相关成纤维细胞，还可通过VEGF、肿瘤相关巨噬细胞促进肿瘤血管生成。CXCL1诱导骨髓来源的抑制性细胞（MDSC）和肿瘤相关巨噬细胞（TAM）在肿瘤组织中的聚集，减少T细胞浸润，从而促进肿瘤细胞发送免疫逃逸。临床上，CXCL1不仅可用于恶性肿瘤辅助诊断和判断患者预后，更可能成为肿瘤生物治疗的干预靶点，对于CXCL1及其受体的调控可能会成为特异性治疗恶性肿瘤的一种重要策略。

Notch signaling in bone marrow mesenchymal stem cells induced by li-popolysaccharide / 中国病理生理杂志

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.

Therapeutic effect of anti-CXCL1 neutralizing antibody on acute ulcerative colitis in mice / 中南大学学报(医学版)

Objective:To evaluate the therapeutic effect of CXCL1 monoclonal antibody on dextra sulfate sodium (DSS)-induced acute ulcerative colitis (UC) in mice,and to elucidate its effect on the expressions ofTNF-α,IFN-γ,,IL-17 and IL-10 as well as neutrophil infiltration.Methods:Female BALB/c mice were randomly divided into a normal group (DSS-),a disease group (DSS+saline),an anti-CXCL1 antibody group (DSS+anti-CXCL1 Ab) and a treatment control group (DSS+IgG Ab).The DSS+saline,DSS+anti-CXCL1 Ab and DSS+anti-CXCL1 Ab groups were given 3.5％ DSS solution as drinking water to induce acute intestinal inflammation,while the normal control was given distilled water freely.The DSS+anti-CXCL1 Ab mice were intraperitoneal injected with anti-CXCL1 Ab (4 mg/kg) on the 3rd and 6th day.Same amount of rat IgG Ab was given in the DSS+IgG Ab group.The normal group and the disease group were injected with 0.9％ sodium chloride solution.The value of disease activity index (DAI) and the injury of colorectal tissue were measured.The levels of TNF-α,IFN-γ,IL-10 and IL-17 in colonic tissues of mice were detected by RT-PCR.Myeloperoxidase (MPO),a specific marker of neutrophils was measured by immunohistochemistry.Results:Compared with the normal control group,DAI score and colorectal injury score in the disease group were significantly increased,but the DAI and colorectal in the mice with acute ulcerative colitis tissue damage score were significantly reduced after anti-CXCL1 Ab intervention.Compared with the normal control group,mRNA levels of TNF-α,IFN-γ and IL-17 in the colorectal tissues were significantly elevated (P＜0.05) in the disease group while the IL-10 was decreased;these effects were attenuated by anti-CXCL1 Ab intervention (P＜0.05).Immunohistochemistry showed that the infiltration of neutrophils (MPO+) in the colon tissue was significantly increased in the disease group,while the anti-CXCL1 Ab treatment could significantly reduce the neutrophil infiltration in colon tissue (P＜0.05).Conclusion:Anti-CXCL1 Ab relieves the progression of DSS-induced acute ulcerative colitis by suppressing proinflammatory expression and neutrophil infiltration.

Clinical validation of multiple biomarkers suspension array technology for ovarian cancer / 中华妇产科杂志

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.

Construction of pEGFP-C1-CXCL1 plasmid and detection of its expression in eukaryotic cells / 军事医学

Objective To construct the pEGFP-C1-CXCL1 eukaryotic expression vector and to investigate the effect of CXCL1 on the proliferation of HepG2 cells under endoplasmic reticulum stress ( ERS).Methods Fragments of CXCL1 were obtained from the cDNA library of HepG2 cells before CXCL1 was cloned into a pEGFP-C1 vector for a recombinant plasmid pEGFP-C1-CXCL1 which was screened and identified by PCR and sequence alignment .Then,the recombinant plas-mid of pEGFP-C1-CXCL1 was transfected into human 293 T cell line and the expression of CXCL 1 was detected by fluores-cence microscopy and Western blotting.pEGFP-C1-CXCL1was furhter transfected into HepG2 cells, and CCK8 was used to detect the inhibitory effect of CXCL1 on tumor proliferation induced by TM in hepatocellular carcinoma .Results pEGFP-C1-CXCL1 was vertified by sequencing analysis .Fluorescence microscopy showed that pEGFP-C1-CXCL1 was transfected into 293T.CXCL1 expression was detected by Western blotting .CCK8 showed that TM inhibited tumor proliferation , while overexpression of CXCL1 decreased the inhabitory rate on cell proliferation of HepG 2 cells under ER stress compared to pEGFP-C1 group and the control group .Conclusion A recombinant pEGFP-C1-CXCL1 plasmid is successfully constructed that can be expressed stably in human 293T cells.Overexpression of CXCL1 can effectively reduce the inhabitory rate of HCC cells induced by the ER stress.

High CXCL1 level correlates with poor prognosis of HBV-associated hepatocellular carcinoma after liver transplantation / 中华普通外科杂志

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.

Effects of stromal cell-derived factor-1 on proliferation,migration,and odontoblastic differentiation of human dental pulp stem cells / 北京大学学报(医学版)

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.

Chemokine CXCL12 and its receptor CXCR4 expression and significance in hypopharynx cancer / 中国医师杂志

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.

Intrathecal injection of oxycodone hydrochloride reduces neuropathic pain in rat model of chronic constriction injury via inhibition of microglial c-JNK/CXCL1 signal / 中国医师杂志

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.

Chemokine (C-X-C Motif) Ligand 1 (CXCL1) Expression in the Minor Salivary Glands of Sjögren's Syndrome Patients

OBJECTIVE: To evaluate the laboratory and clinical manifestations of Sjögren's syndrome (SS) association with chemokine (C-X-C motif) ligand 1 (CXCL1) expression in the ductal and acinar salivary gland epithelial cells (SGEC) of the minor salivary glands. METHODS: The sociodemographic data of 106 SS patients was obtained, and the glandular and extraglandular manifestations of the disease documented. The minor salivary glands were biopsied and the laboratory findings analyzed. European League Against Rheumatism SS disease activity index (ESSDAI) and SS disease damage index (SSDDI) scores were obtained during biopsy. An immunohistochemical approach was used to define the expression of CXCL1 in the salivary glands. RESULTS: Of 106 patients, the minor salivary glands of 22 patients (20.7%) stained positively for CXCL1. Such CXCL1-positive patients exhibited higher ESSDAI scores at the time of biopsy than the CXCL1-negative patients (3.86±2.27 vs. 2.64±1.62, p=0.015). Lymphadenopathy was more frequently observed in CXCL1-positive patients, compared with CXCL1-negative patients (31.8% vs. 9.5%, p=0.014). No differences between groups were identified in terms of sociodemographic characteristics, laboratory data, or the extent of the glandular manifestation of SS. CONCLUSION: The expression of CXCL1 within the ductal and acinar SGEC of SS patients is associated with lymphadenopathy and elevated clinical disease activity. CXCL1 may play an important role in the disease activity and prognosis of SS.

Expression and the prognosis value of CXCL1 2/CXCR4 in adenoid cystic carcinoma / 实用口腔医学杂志

Objective:To investigate the expression of CXCL1 2 and CXCR4 in adenoid cystic carcinoma(ACC)and to explore its re-lationship with clinicopathologic characteristics and prognosis of the patients.Methods:The expression of CXCL1 2 and CXCR4 in af-fected tissue was detected immunohistochemically in 62 cases of ACC.Both of the two factors and clinicalpathology factors were pro-cessed in accordance with the Kaplan-Meier method and the COX regression model.Results:The positive rates of CXCL1 2 and CX-CR4 expression were 54.8%(34/62)and 77.42%(48/62)respectively.Patients with the 2 factor expression had a shorter survival time than those without them(P<0.05).Multivariate analysis revealed that CXCR4 expression,clinical stage,histological differentia-tion and metastasis/recurrence were independent risk factors for the prognosis of ACC patients.Conclusion:The expression of CXCR4 may be correlated with the malignancy of ACC.CXCR4 expression,clinical stage,metastasis/recurrence and histological differentia-tion can indicate the prognosis of ACC patients.

The changes of GRO-α and VEGF in the peritoneal fluid and their relationship in the patients of endometriosis / 中国医师杂志

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.