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OBJECTIVE@#To explore the value of interferon-inducible protein 10 (IP-10) in the auxiliary diagnosis of tuberculosis and the judgment of the severity of disease.@*METHODS@#From February, 2013 to February, 2017, a total of 193 patients with TB admitted in our hospital and 84 healthy control subjects were recruited consecutively. The peripheral blood plasma levels of interferon-γ (IFN-γ) and IP-10 were detected using liquid phase chip (Luminex) technique. According to the number of lung fields affected by TB, the patients were divided into group A (with lesions in 1-2 lung fields), group B (3-4 lung fields) and group C (5-6 lung fields), The expressions of IFN-γ and IP-10 in 3 groups were compared.@*RESULTS@#The plasma levels of IP-10 were significantly higher in TB patients than in the control subjects ( < 0.05), but IFN-γ levels were comparable between the two groups ( > 0.05). Among the TB patients, plasma IP-10 levels was the highest in group C ( < 0.05), and IFN-γ levels did not differ significantly among the 3 groups ( > 0.05).@*CONCLUSIONS@#Plasma IP-10 has a certain reference value in the auxiliary diagnosis of active tuberculosis and the judgment of the severity of the disease.
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Humans , Antigens, Bacterial , Biomarkers , Blood , Chemokine CXCL10 , Blood , Tuberculosis, Pulmonary , Blood , DiagnosisABSTRACT
Objective To evaluate early diagnostic value of matrix metalloproteinase 7 (MMP-7) and interferon inducible protein (IP-10) for rheumatoid arthritis-associated interstitial lung disease (RA-ILD).Methods Clinical and laboratory data and serum samples of patients with RA between March 2015 and June 2016 in our hospital were collected.Patients were subclassified as RA-without ILD,RA-with ILD,or RA-advanced ILD based on high-resolution computed tomography scans of the chest.Enzyme linked immunosorbent assay (ELISA) was used to assess MMP-7 and IP-10 in the serum of each group.The correlation between the two biomarkers and pulmonary function,the occurrence of ILD and other laboratory indexes were analyzed.Comparison of measurement data between the 3 groups was performed by single factor variance analysis,while count data was compared by Chi square test;the risk factors were analyzed by Logistic regression,receiver operating characteristic curve (ROC curve) was used to evaluate the diagnostic efficiency,correlation analysis was performed by Spearman correlation analysis.All statistical analysis was performed by Statistical Product and Service Solutions (SPSS) 19.0 statistical software.Results Serum levels of MMP-7 and IP-10 in RA-advanced ILD group and RA-mild ILD group were significantly higher than that of RA-without ILD group,the level of MMP-7 was [(1.9±3.0) ng/ml,(1.7±2.4) ng/ml,(0.2±0.2) ng/ml in turn,P＜0.05],IP-10 level was [(245± 394) pg/ml,(270±384) pg/ml,(38±26) pg/ml in turn,P＜0.05].And IP-10 level was significantly correlated with RA-ILD in both unadjusted and adjusted Logistic regression analyses,the values of OR were 1.024 and 1.023 respectively (P＜0.05).The ROC curves were plotted with MMP-7 and IP-10 respectively.The area under the curve (AUC) was 0.69 and 0.78 respectively.The AUC of combined detection increased to 0.83.Conclusion Levels of MMP-7 and IP-10 are elevated in the serum of RA patients with ILD,whether mild or advanced,supporting their value as pathogenically relevant biomarkers that may contribute to noninvasive detection of RA-ILD,and the combined estimate of them helps to improve the effectiveness of early diagnosis.
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Objective To investigate the predictive value of dynamic changes of interferon-inducible protein-10 (IP-10) expression in the peripheral blood mononuclear cells and the model for end-stage 1iver disease (MELD) scores for short-term mortality in hepatitis B virus related acute-on-chronic liver failure (HBV-ACLF) patients .Methods Eighty patients with HBV-ACLF admitted to the Affiliated Hospital of Hubei College of Arts and Sciences from October 2013 to August 2015 were selected .During 3 months of follow-up ,33 patients died and 47 survived .The expression level of IP-10 and MELD score of two groups were measured at admission and week 1 and week 2 after treatment .The means between two groups were compared .Accuracy of predicting short-term mortality was performed by area under receiver operating characteristic curve (AUC) .Multivariate logistic regression analysis and Kaplan-Meier survival curve were used to analyze the effect of IP-10 expression and MELD score on the mortality of HBV-ACLF patients . Results The expressions of IP-10 at admission and at week 1 and 2 after treatment in the death group were 1 .095 ± 0 .202 ,1 .071 ± 0 .181 ,and 1 .078 ± 0 .198 ,respectively ,those in the survival group were 0 .894 ± 0 .181 ,0 .770 ± 0 .153 ,and 0 .732 ± 0 .137 ,respectively ,which were significantly different (t =4 .66 ,8 .02 and 9 .27 ,respectively ,all P < 0 .01) .The MELD scores at admission and at week 1 and 2 after treatment in the death group were 26 .70 ± 5 .50 ,27 .39 ± 6 .24 ,and 28 .64 ± 6 .44 ,respectively , those in the survival group were 23 .89 ± 4 .41 ,21 .57 ± 4 .68 ,and 18 .87 ± 3 .92 ,respectively ,which were significantly different (t= 2 .53 ,4 .77 and 8 .42 ,respectively ,all P< 0 .01) .Analysis of variance showed that the MELD score and IP-10 expression in the survival group at admission were significantly higher than those at week 1 and week 2 after treatment (F= 13 .464 and 15 .711 ,respectively ,both P< 0 .01) ,while there were no significant differences in the death group (F = 0 .129 and 0 .864 ,respectively ,both P >0 .05) .The AUC of IP-10 at week 2 after treatment was 0 .935 ,that of MELD score was 0 .903 (Z =0 .788 ,P= 0 .045) ,while there was no significant difference of AUC between week 1 and week 2 (0 .935 vs 0 .909 ,Z = 0 .640 ,P> 0 .05) .In addition ,the AUC of IP-10 level at week 1 and MELD score at week 2 after treatment showed no significant difference (0 .909 vs 0 .903 ,Z = 0 .133 , P > 0 .05) .Logistic multivariate regression analysis showed that IP-10 ≥ 0 .902 at week 1 ,MELD ≥ 22 .5 and IP-10 ≥ 0 .846 at week 2 were independent risk factors for death (OR= 11 .29 ,6 .60 ,and 15 .27 ,respectively ;95％ CI =1 .06 - 119 .74 ,1 .27 - 34 .26 ,and 1 .39 - 167 .62 ,respectively ;all P< 0 .05) .Conclusion The dynamic monitor of both IP-10 levels and MELD scores may have greater value in predicting prognosis of patients with HBV-ACLF .
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The pathogenesis of liver failure is extremely complex. In recent years, the role of chemokines in viral hepatitis has been widely concerned. A large number of studies have confirmed that the abnormal expression of chemokines is closely related to the process of viral hepatitis and other types of hepatitis. Interferon-inducible protein-10 (IP-10) is a good indicator of the specificity and sensitivity of inflammatory liver injury. In this paper, IP-10 in different types of liver injury in the pathogenesis of research progress were reviewed to provide valuable indicators for clinical diagnosis, treatment strategy and prognosis evaluation of liver injury.
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HBV-associated acute-on-chronic liver failure is prevalent in mainland China.The prognosis of HBV-ACLF is poor.The mortality of HBV-ACLF is approximately 80％.Therefore,a prognostic indicator was needed in order to allow us to intervene as soon as possible.The model for end-stage liver disease (MELD) scoring system is widely used to predict the prognosis of liver failure.However,the assessment is too complex to restrict its application.This study aimed to investigate the expression ofIP-10 in peripheral blood mononuclear cells (PBMC),in order to explore the relationship between the expression and prognosis of patients with HBV-ACLF.The mRNA level of IP-10 in PBMCs were analyzed in 80 patients with HBV-ACLF,40 patients with chronic hepatitis B (CHB)and 40 healthy people by fluorescent quantitative PCR.IP-10 mRNA level was significantly higher in the HBV-ACLF group than in the other two groups (P＜0.01).Group with MELD score below 30 had lower IP-10 mRNA level than group with MELD score over 30 (P＜0.05).The IP-10 mRNA level in PBMCs in positive group was higher than that in negative group (P＜0.01).With a threshold of 0.925,the area under the receiver operating characteristic (ROC) curves was 0.815.These findings suggest that assessment of IP-10 mRNA level in the PBMCs would be helpful for evaluating the disease severity and prognosis in patients with HBV-ACLF.
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Objective To investigate the correlations of expression of interferon inducible protein 10 ( IP-10) mRNA in peripheral blood mononuclear cells ( PBMCs) with serum levels of HBsAg and HBV DNA in patients with HBV-related acute on chronic liver failure ( HBV-ACLF ) , and to assess its value in predicting disease prognosis .Methods Eighty patients with HBV-ACLF, 60 patients with chronic hepatitis B (CHB), and 25 healthy subjects were enrolled during October 2013 and February 2015.IP-10 mRNA in PBMCs was measured by real time quantitative polymerase chain reaction ( RT-qPCR) , and the difference in IP-10 mRNA expression among three groups was compared by ANOVA .Serum levels of HBsAg and HBV DNA were also measured in patients with HBV-ACLF, and Pearson correlation test was performed to analyze the correlations of IP-10 mRNA with HBsAg and HBV DNA levels .t test was used to analyze the differences in IP-10 mRNA, HBsAg and HBV DNA levels between fatal and surviving cases after 3-month entecavir therapy in HBV-ACLF group.Receiver operating characteristic curves ( ROC) was used to evaluate the prognostic value of IP-10 mRNA, HBsAg and HBV DNA in HBV-ACLF patients .Results IP-10 mRNA level in HBV-ACLF group was 0.998 ±0.186, which was higher than those in CHB patients and healthy controls (0.641 ±0.083 and 0.412 ±0.062, t=3.841 and 16.661, P<0.01).Pearson correlation analysis showed that IP-10 mRNA level was negatively correlated with HBsAg level in HBV-ACLF patients (r=-0.576, P<0.01), but positively correlated with HBV DNA level (r=0.547, P<0.01).After 3-month entecavir therapy , IP-10 mRNA level in surviving cases of HBV-ACLF group was 0.894 ±0.164, which was lower than that in fatal cases ( 1.103 ±0.177, t =-4.328, P <0.01 ); HBsAg level in surviving cases was higher than that in fatal cases (3.303 ±0.565 vs.2.605 ±0.844, t =3.251, P<0.01).The area under ROC of IP-10 mRNA in evaluating prognosis of HBV-ACLF was 0.820, which was higher than those of HBsAg (0.663) and HBV DNA (0.570).Conclusions IP-10 mRNA in patients with HBV-ACLF is over-expressed and is correlated with HBsAg and HBV DNA levels .It may be used for predicting the prognosis of patients with HBV-ACLF.
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Objective To investigate the potential role of Lycium bararum polysaccharide (LBP) with or without interferon -inducible protein 10 ( CXCL10) in inducing dendritic cells ( DC) functional maturation by monitoring the alteration of cytokines for inducing DC maturation in peripheral blood and by detecting the expression of S-100 protein in tumor tissue, thus to reveal its mechanism of inhibiting experimental liver cancer. Methods H22 bearing mice model was established. The mice were randomized into model group, LBP group (50 mg/kg, ig), CXCL10 (right axillary subcutaneous injection of 15 μg/kg), LBP + CXCL10 group (LBP 50 mg/kg, ig, and right axillary subcutaneous injection of CXCL10 15 μg/kg), 5- fluorouracil (5FU) group ( intraperitoneal injection of 12mg/kg) , 12 mice in each group. The mice were administered the corresponding medicine once a day. After treatment for 2 continuous weeks, blood was sampled from infraorbital vein, and the tumor mass, spleen, thymus were extracted for the calculation of anti-tumor rate, thymus index and spleen index separately . The mRNA expression levels of interleukin 12 (IL-12) and tumor necrosis factor-α (TNF-α) in peripheral blood were detected by fluorescence quantitative PCR, the expression of S-100 protein in tumor tissues was detected by immunohistochemical assay. Results Compared with the model group, tumor growth in LBP group and LBP+CXCL10 group was obviously inhibited, and tumor-inhibitory rate was 55.90%, 50.91%, respectively. Meanwhile, the mRNA expression level of IL-12 was 2.94 folds higher in LBP group and 3.39 folds higher in LBP + CXCL10 group, and TNF-α mRNA expression level was 1.55 folds higher in LBP group and 4.74 folds higher in LBP+CXCL10 group than the model group, the differences being statistical significant ( P<0.05 or P<0.01). Results of immunohistochemical assay showed that S-100+DC number in LBP group and LBP+CXCL10 group was larger than that in the model group (P<0.05 ). Conclusion LBP and LBP+CXCL10 exert significant effect on inhibiting experimental liver cancer. The mechanism may be related with inducing the secretion of IL-12 and TNF-α, which plays a key role in inducing DC maturation, and with the increase of the number of DC in tumor microenvironment.
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Objective To investigate the expression of interferon inducible protein-10 (IP-10) in peripheral blood mononuclear cells (PBMC) of patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF), and its correlation with disease severity.Methods Eighty patients with HBV-ACLF, 60 patients with chronic hepatitis B (CHB), and 25 healthy controls were enrolled from the Affiliated Hospital of Hubei University of Arts and Science during October 2013 and February 2015.IP-10 mRNA in PBMC was measured by real time quantitative PCR.Independent sample t test was used to analyze the difference in IP-10 mRNA expression between HBV-ACLF patients with model for end-stage liver disease (MELD) ＜ 30 and ≥30, and Pearson correlation test was performed to analyze the correlations of IP-10 mRNA expression with alanine aminotransferase (ALT), total bilirubin (TBil) and international normalized ratio (INR).Results The expressions of IP-10 mRNA in HBV-ACLF patients was 1.00 ± 0.19, which was higher than those in CHB patients and healthy controls (0.64 ± 0.08 and 0.41 ± 0.06, t =3.841 and 16.661, all P ＜ 0.01).The expression of IP-10 mRNA in HBV-ACLF patients with MELD ＜ 30 was 0.96 ±0.19, which was lower than that in patients with MELD ≥ 30 (1.14 ± 0.21, t =-2.283, P ＜0.05).Pearson correlation analysis showed that IP-10 mRNA level in HBV-ACLF patients was positively correlated with ALT, TBil and INR (r =0.697, 0.738 and 0.775, all P ＜ 0.01).Conclusion IP-10 mRNA is over-expressed in PBMC of patients with HBV-ACLF, and it is correlated with disease severity, which suggests that IP-10 may play an important role in the progression of liver failure.
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BackgroundIt has been proved that as a chemokine,interferon-inducible protein-10(IP-10)can regulate the immuno-inflammatory reaction.Some new researches showed that IP-10 also played role in regulating the neovascular vessel formation.Corneal neovascularization (CNV) is associated with multiple cellular factors,but its mechanism is below clear.Objective The present study was to address the roles of exogenous mouse IP-10 in alkali burn-induced CNV.Methods Eighty-two SPF BALB/c mice were used in this experiment and grouped according to random number table.The corneal alkali burn models were established by putting the filter paper with 1 mol/L NaOH at the central corneas of the left eyes for 40 seconds.10 mg/L IP-10 was topically administered from the first day or 14 days after modeling in the early intervene group( 10 eyes)or middle-late intervene group(5 eyes).CNV area was measured as a percentage of whole cornea.0.2％ sodium hyaluronate(HA) as vehicle was utilized in the model control group.Angiogenic factor expression in corneal tissue in the early intervene group was quantified by reverse transcriptase polymerase chain reaction (RT-PCR)and compared with model control group.All animal experiments were performed in accordance with the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research and complied with the standards of Guidelines for the Care and Use of Laboratory Animals of Soochow University. ResultsThe CNV percentage was(88.67±10.22) ％ in the model control mice,showing a significant increase in comparison with that of IP-10 early intervene group (70.06±12.21)％ (t=3.77,P=0.00).In 21 days after corneal alkali burn,the CNV percentage was(87.33±13.47)％ in the model control mice,and that of the IP-10 middle-late intervene group was ( 86.56± 12.47 ) ％ without significant difference between them ( t =1.26,P＞0.05 ).Two days or four days after IP-10 early intervene,the expressions of chemokine receptor type 3 ( CXCR3 ) in corneal tissue were significant higher than model control group( t =3.13,3.07,P＜0.05 ),but the expressions of vascular endothelial growth factor (VEGF) in cornea were lowed ( t =5.99,6.27,P＜0.01),and so were transforming growth factor-β1 (TGF-β1) (t =8.50,P＜0.01;t =4.53,P＜0.05).Conclusions The early topical administration of the exogenous mouse IP-10 can inhibit CNV by up-regulating CXCR3 expression and down-regulating VEGF and TGF-β1 expression in cornea.However,middle-later usage of the IP-10 is uneffective.
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Objective To investigate the expressions of interferon γ(IFNγ), interferon inducible protein-10(IP-10). chemokine receptor CXCR3 and their significance in infection-associated hemophagocytic syndrome(IAHPS). Methods Forty-three patients with IAHPS, 35 infection patients without HPS and 25 healthy controls were included in the study. The serum IFNγ and IP-10 levels were measured by enzyme linked immunosorbent assay(ELISA). The expression of CXCR3 on cell surface of CD_4~+ and CD_8~+ T cells in peripheral blood was determined by flow cytometry. SPSS 13.0 was used for data processing, and independent-sample t test was performed to compare the differences among the groups. Results The serum IFNγ and IP-10 levels in patients with IAHPS were( 608±135) pmol/L and(939±141) pmol/L respectively, which were significantly higher than those in without HPS group[(154±45) pmol/L and (385±119) pmol/L, t=4.97 and 4.02, P<0.05] and healthy controls[(56±18) pmol/L and (248±98) pmol/L, t=5.27 and 4.77, P<0.05]. The expressions of CXCR3 on CD_4~+ and CD_8~+ T cells in IAHPS group were (35±11)% and (23±8)% respectively, which were significantly higher than those in without HPS group[(24±7)% and (16±7)%, t=3.12 and 3.62, P<0.05] and healthy controls[(20±6)% and (12±5)%, t=4.46 and 4.93, P<0.05]. Conclusion The expressions of IFNγ, IP-10 and CXCR3 are increased significantly in patients with IAHPS, which may be related to the disease pathogenesis.
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BACKGROUND/AIMS: Recent studies have shown that serum interferon gamma-inducible protein-10 (IP-10) concentration decreased after pegylated interferon and ribavirin therapy, and was associated with a sustained virologic response (SVR). The aim of this study was to investigate the clinical significance of the pretreatment IP-10 level and change in serum IP-10 level between 1 month before and after treatment and its association with various virologic responses in patients having chronic hepatitis C (CHC) with genotype 1 undergoing pegylated interferon and ribavirin therapy. METHODS: Thirty-six patients having CHC with genotype I undergoing pegylated interferon and ribavirin therapy who had available stored sera 1 month before and after treatment were enrolled retrospectively. Serum IP-10 levels were measured by ELISA. Serum HCV RNA was measured by RT-PCR (detection limit0.05). The change in serum IP-10 between 1 month before and after treatment had no clinical meaning based on various virologic responses (p>0.05). CONCLUSIONS: The level of pretreatment IP-10 and change in IP-10 level between 1 month before and after treatment were not predictive factors of a SVR. Additional large-scale studies to determine the SVR-predicting role of serum IP-10 levels in patients with CHC are needed.
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Humans , Enzyme-Linked Immunosorbent Assay , Genotype , Hepatitis C, Chronic , Hepatitis, Chronic , Interferons , Retrospective Studies , Ribavirin , RNAABSTRACT
Objective To explore the relationship between serum levels of interferon-inducible protein-10 (IP-10), interferon-γ (IFN-γ) and hepatic inflammatory reaction, disease progression in patients with severe hepatitis B (SHB). Methods Sera of 40 patients with SHB at time of admission,at the beginning of single plasma exchange (PE), at time of PE completion and 5 days after PE. The SHB patients were divided into improved group and aggravated group. And 20 patients with chronic hepatitis B (CHB) and 20 healthy controls were enrolled in this study. Serum levels of IP-10, IFN-γand tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). Results The serum levels of IP-10 in patients with SHB and CHB on admission were (683.6 174.6)ng/L and (216.1 102.9)ng/L, respectively, which were notably higher than those in healthy controls [(107.6 55.8)ng/L F=9.036, both P＜0. 01],and those in patients with SHB was significantly higher than that in patients with CHB (P＜0. 01). The serum level of IFN-γ in patients with SHB and CHB on admission were (19. 8 8. 8) ng/L and (16. 7 7. 8) ng/L,respectively, which were significantly higher than those in healthy controls [(2.6 1.2) ng/L F=9. 288, both P＜0. 01]. The serum level of IP-10 and IFN-γ were both positively correlated with TNF α (r=0. 366 and r=0. 365, respectively;P＜0.05) and both negatively correlated with prothrombinase activity (r=-0.401 and r=-0.350, respectively;P＜0.05), but not correlated with serum total bilirubin(r=0. 223 and r=0. 219, respectively;P＞0.05). The serum level of IP-10 and IFN-γ were positively correlated ( r= 0. 602 ; P= 0. 000 ). On day 5 after PE, serum level of IP-10 in patients with SHB was significantly decreased compared with that'in patients before PE (t= 8. 947, P＜0.01 in improved group;t=4. 121, P＜0.05 in aggravated group) and that in aggravated group was significantly higher than improved group (t=7.862, P＜0.01). But serum level of IFN-γ was not decreased significantly (t=0. 491, P＞0.05). Conclusions IP-10 and IFN-γ are involved in the hepatic immunopathological mechanism. Serum level of IP-10 is correlated with the severity of hepatic inflammatory injury and IP-10 could reflect the progression and development of disease in patients with SHB.
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Objective To investigate the significance of serum IP-10 levels in different types of diabetic patients. Methods Serum IP-10 level in 78 cases with type 1 diabetes,49 cases with type 2 diabetes and 33 cases of healthy controls was measured with ELISA assays. Type 1 diabetic patients were divided into classic type 1 diabetes (n=39) and latent autoimmune diabetes in adults (LADA,n=39) according to their disease process,and they were also divided into autoimmune type 1 diabetes (n=58) and idiopathic type 1 diabetes (n=20).Type 2 diabetic patients,according to carotid intima-media thickness (IMT),were divided into patients with (n=24) or without atherosclerosis (n=25). Results 1) Serum IP-10 levels were not statistically different among type 1 diabetes,type 2 diabetes and healthy controls. 2) Serum IP-10 concentrations in autoimmune type 1 diabetes with positive islet autoantibody were higher than those in healthy controls(184.96?104.48pg/ml vs 146.10?74.61pg/ml,P=0.03);while there were no difference in type 1 diabetes with variable disease durations. 3) No difference in IP-10 levels was found between type 2 patients with(173.5?69.6pg/ml) and without atherosclerosis (188.5?79.7 pg/ml). Conclusions Serum IP-10 level is augmented in autoimmune type 1 diabetes.
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Objective To observe the expressions of the chemokine receptor CXCR3 and its ligand IFN-?-inducible protein-10(IP-10) in lung tissue of asthmatic model mice and to explore further the effect of dexamethasone(DEX)and bacillus Calmette-Guerin vaccine(BCG)on the expressions of CXCR3 and IP-10.Methods Forty Kunming mice were randomly divided into 4 groups:asthmatic group,DEX group,BCG group and control group,10 in each group.Mice were sensitized and challenged with ovalbumin (OVA) to establish asthmatic models.Ten mice in DEX group were administrated DEX (2 mg/kg) by abdomen injection 30 min before challenge (DEX group).Ten mice in BCG group were injected BCG(0.025 mg) intradermally 7,3,and 1 days before sensitization.The mice in control group mice were treated with 9 g/L saline instead of OVA.Mice of each group were executed 24 hours after the final challenge.Their lung tissue were paraffin embedded,sliced and stained by HE.The expressions of CXCR3 and IP-10 protein in lung tissue of mice were detected by immunohistochemistry.Results Distinct differences were discovered in the expressions of CXCR3(F=4.602 P=0.008) and IP-10(F=4.207 P=0.012) among the 4 groups.The expressions of CXCR3 and IP-10 in lung tissue of mice in asthmatic group were significantly higher than those in control group (Pa=0.002),while that in DEX group were significantly lower than that in asthmatic group (P=0.029,0.019).There were no significant difference between the BCG group and asthmatic group.Conclusions The chemokine receptor CXCR3 and its ligand IP-10 play an important roles in mechanisms in the pathogenesis of asthma.DEX and BCG can interfere the expressions of CXCR3 and IP-10 in varying degress.
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AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.
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Objective To clone the 5′ non-coding region (NCR) of human interferon-?-inducible protein 10(IP-10), and to identify the transcriptional activity of IP-10 promoter induced by lipopolysaccharide (LPS) in human umbilical vein endothelial cells (HUVEC). Methods Genomic DNA of lymphocytes was isolated from the human blood. With above DNA as the template, the 5'NCR of human IP-10 was amplified by nest polymerase chain reaction (PCR) method. Then, the IP-10 promoter was cloned into luciferase reporter vector, pGL3. The recombined vector was transfected into HUVEC, and then the activity of the luciferase was determined after the cells were stimulated by LPS. Results Human IP-10 promoter was obtained and the pGL3/IP-10 was successfully constructed. Moreover, the activity of luciferase driven by human IP-10 promoter was observed to obviously increase in the HUVEC stimulated by LPS. Conclusion We successfully cloned human IP-10 promoter, constructed luciferase reporter vector driven by the human IP-10 promoter, and confirmed that high transcriptional activity of human IP-10 promoter was induced by LPS in HUVEC. The results supplied an experimental base for the further study of the transcriptional regulation of human IP-10.