Oncolytic virus is an emerging anti-cancer strategy. However, extracellular matrix(ECM), as a physical barrier, limits virus spread within the tumor. To overcome the obstacle, we constructed a recombinant Newcastle di...Oncolytic virus is an emerging anti-cancer strategy. However, extracellular matrix(ECM), as a physical barrier, limits virus spread within the tumor. To overcome the obstacle, we constructed a recombinant Newcastle disease virus(NDV) expressing matrix metalloproteinase(MMP8)(NDV-MMP8) using with reverse genetic technology. In vitro, NDV-MMP8 was identified and verified by WB and ELISA. Cell viability was detected by CCK-8 assay. In vivo, we established two liver cancer xenograft models. NDV-MMP8 was injected into the tumor to observe the tumor volume and survival of mice. The changes in extracellular matrix were observed by Masson’s trichrome staining. Virus expression in tumor tissues was detected by immunofluorescence assay. The virus titer in tumor tissues was detected by TCID50. Histopathological changes were detected by hematoxylin and eosin(HE) and terminal deoxynucleotidyl transferase d UTP nick end labeling(TUNEL) staining. Intratumoral administration of NDV-MMP8 can effectively degrade ECM, promote the spread of the virus within the tumor, and reduce tumor growth rate. Therefore, the method of increasing intratumoral virus accumulation by degradation of the ECM to enhance the oncolytic effect has great potential for clinical application.展开更多
Objective To study the clinical role of the variation of serum matrix metalloproteinase-8 (MMP-8) concentration in patients with acute coronary syndrome (ACS). Methods ELISA method was adopted to detect serum MMP-8 co...Objective To study the clinical role of the variation of serum matrix metalloproteinase-8 (MMP-8) concentration in patients with acute coronary syndrome (ACS). Methods ELISA method was adopted to detect serum MMP-8 concentration and to observe concentration’s differences and features among 80 selected ACS cases (43 acute myocardial infarction and 37 unstable angina pectoris), 43 stable angina pectoris (SAP) cases and 37 control cases. And meanwhile the atherosclerosis risk factors of each case, such as age, sex, hypertension, body mass index, smoking, family history, diabetes, and hyperlipidemia were collected and analyzed as a whole. Results First, serum MMP-8 concentration reached the highest point in ACS, and there was significant difference between SAP and control groups (P<0.01). Second, serum MMP-8 in AMI was much higher than that in UAP with significant difference (P<0.01). There was no difference between UAP and SAP groups (P>0.05). Third, Logistic regression analysis revealed that serum MMP-8 concentration might be the indicator of ACS (B=4.493, P=0.000), particularly, that of AMI (B=9.961, P=0.000). Fourth, linear correlation and linear regression analysis found that only neutrophil was likely to influence serum MMP-8 concentration (r=0.274, P=0.001). Fifth, in the diagnosis of ACS, the area under ROC curve of MMP-8 was 0.785, the sensitivity and specificity were 68.6% and 76.5%, respectively. Conclusion ① Serum MMP-8 concentration has close relationship with the occurrence of ACS, particularly with AMI; ② Serum MMP-8 concentration may be one of the predicting indicators of ACS and particularly of AMI; ③ Neutrophil may be correlated with serum MMP-8 concentration; ④ MMP-8 is of somewhat valuable in diagnosing ACS.展开更多
Background Sepsis-induced myocardial injury （SIMI） is caused by a variety of mechanisms. The aim of the study is to investigate the effects of metalloproteinase-8 （MMP-8） on SIMI and its mechanisms in rats. Method...Background Sepsis-induced myocardial injury （SIMI） is caused by a variety of mechanisms. The aim of the study is to investigate the effects of metalloproteinase-8 （MMP-8） on SIMI and its mechanisms in rats. Methods Forty male Sprague Dawley rats were randomly divided into four groups： MMP-8 inhibitor （M81）, dexamethasone （DEX）, sepsis, and sham groups. The sepsis model was established by cecal ligation and puncture （CLP）. Rats in the M81 group immediately received an intraperitoneal injection of M81 （0.1 mg/kg） after CLP. Rats in the DEX group immediately received an intraperitoneal （IP） injection of DEX （2 mg/kg）. Rats in the sepsis and sham groups received intraperitoneal injections of normal saline. Rats were sacrificed 12 hours after CLP. Paraffin sections were stained with hematoxylin and eosin to observe the myocardium. The myocardial ultrastructure was observed with transmission electron microscopy. MMP-8, tumor necrosis factor-Q （TNF-a）, and interleukin-113 （IL-113） were detected by immunohistochemistry. The expression of MMP-8 was measured by Western blotting. TNF-a and IL-113 levels in serum and myocardial tissue were determined by enzyme-linked immunosorbent assay. Results Compared with the sham group, the myocardium in the sepsis group was seriously injured. MMP-8, TNF-α and IL-1β expression was higher in the sepsis group than in the sham group, Treatment with M81 or DEX, however, attenuated sepsis induced histopathological changes in the heart, and was associated with significant reductions in serum and myocardial levels of TNF-a and IL-113 （P 〈0.05）. M81 significantly inhibited MMP-8 expression in myocardial tissue （P 〈0.05）. In addition, treatment with DEX was not associated with a change in myocardial levels of MMP-8 （P 〉0.05）. Conclusion MMP-8 inhibitor attenuated myocardial injury in septic rats, which might be related to reduced expression of TNF-α and IL-1β.展开更多
Periodontitis is an inflammatory disease of the periodontium. Any imbalance between the matrix metalloproteinases （MMPs） secreted by neutrophils and tissue inhibitors initiates the destruction of collagen in gum tis...Periodontitis is an inflammatory disease of the periodontium. Any imbalance between the matrix metalloproteinases （MMPs） secreted by neutrophils and tissue inhibitors initiates the destruction of collagen in gum tissue, leading to chronic periodontitis. This study aimed to correlate salivary levels of MMP-8 and periodontal parameters of chronic periodontitis to establish MMP-8 as a noninvasive marker for the early diagnosis of chronic periodontitis. The study involved 40 subjects visiting the periodontic OPD of Dr. Ziauddin Ahmad Dental College and Hospital, located in Aligarh, U.P., India, from 2011 to 2012. The subjects were divided into two groups： group I consisted of 20 periodontally healthy subjects （controls） while group II consisted of 20 patients with chronic periodontitis. Chronic periodontitis was assessed on the basis of several periodontal parameters, including pocket probing depth （PPD）, clinical attachment level （CAL）, gingival index （GI）, and plaque index （PI）. Around 3 ml of unstimulated and whole expectorated saliva was collected for MMP-8 estimation by ELISA using Quantikine human total MMP-8 immunoassay kits. Data were analyzed using STATISTICA （Windows version 6） software. Salivary MMP-8 levels of groups I and II were 190.91 ± 143.89 ng/ml and 348.26± 202.1 ng/ml, respectively. The MMP-8 levels and periodontal status （PPD, CAL, GI, and PI） of groups I and II showed positive and significant correlations （for PPD, r = 0.63, P 〈 0.001; for CAL, r = 0.54, P 〈 0.001; for GI, r = 0.49, P 〈 0.001; and for PI, r = 0.63, P 〈 0.001）. The results of this study demonstrate elevated concentrations of MMP-8 in individuals with chronic periodontitis.展开更多
Background Nasopharyngeal carcinoma (NPC) shows highly invasive and metastatic features. This study aims to investigate macrophage migration inhibitory factor (MIF)-induced invasion of NPC cells in vitro and the eff...Background Nasopharyngeal carcinoma (NPC) shows highly invasive and metastatic features. This study aims to investigate macrophage migration inhibitory factor (MIF)-induced invasion of NPC cells in vitro and the effects on matrix metalloproteinases (MMPs) and interleukin-8 (IL-8),and to study the mechanism of tumor cell invasion and metastasis in the early stage of NPC. Methods Two nasopharyngeal carcinoma cell lines,CNE-1 and CNE-2,were adopted in this study. The NPC cell invasion and migration were evaluated by microinvasion assay. The variation of expression percentages of MMP2- or MMP9-positive cells was detected by flow cytometry in two cell lines with or without MIF treatment. Western blotting and RT-PCR were used to assay the protein and mRNA expressions of MMP2 and MMP9. The IL-8 concentration secreted by NPC cells was compared with the cells with different treatments using ELISA. Results After treating with MIF for 48 hours,the cell numbers of CNE-1 and CNE-2 which went through the 8-μm filter membrane were increased. Compared with non-MIF treated NPC cells,significant difference could be found both in CNE-1( P =0.005) and CNE-2 cells ( P =0.001) . The percentages of MMP9-positive cells were significantly increased in both CNE-1 [from (28.5±2.5)% to (82.4±3.5)%, P =0.001] and CNE-2 [from (32.8±3.5)% to (86.1±1.6)%, P =0.002]. The relative intensity of MMP9 protein expression was also enhanced in both cell lines (CNE-1: from 83.1±6.0 to 242.9±22.9, P =0.002;CNE-2: from 84.4±4.3 to 278.9±29.7, P =0.003). Correspondingly,the increased MMP9 mRNA expression level was significantly detectable in both cell lines.The concentration of IL-8 in the supernatant of CNE-2 was higher [(1201.8±593.3) pg/ml] after treatment. It was also remarkably higher than that in the supernatant of CNE-2 without treatment ( P =0.026). However,there was no significant difference in the concentration variation of IL-8 in CNE-1 ( P =0.581), while the IL-8 mRNA level was only enhanced in CNE-2. Conclusions MIF can induce potent invasion of NPC cell lines in vitro , and the infiltrating lymphocytes in NPC might be responsible for the invasion and metastasis of tumor cells. MIF cytokine which is secreted by these infiltrating lymphocytes might contribute to the invasion as well as metastasis of NPC in the early stages by induction of MMP9 and IL-8 in an indirect pathway.展开更多
基金supported by the Scientific and Technological Innovation Major Base of Guangxi (No. 2018–15-Z04)Guangxi Key Research and Development Project (No. AB20117001)+1 种基金Guangxi Science and Technology Bases and Talent Special Project (No. AD17129062)Guangxi Natural Science Foundation (No. 2018JJA140524)。
文摘Oncolytic virus is an emerging anti-cancer strategy. However, extracellular matrix(ECM), as a physical barrier, limits virus spread within the tumor. To overcome the obstacle, we constructed a recombinant Newcastle disease virus(NDV) expressing matrix metalloproteinase(MMP8)(NDV-MMP8) using with reverse genetic technology. In vitro, NDV-MMP8 was identified and verified by WB and ELISA. Cell viability was detected by CCK-8 assay. In vivo, we established two liver cancer xenograft models. NDV-MMP8 was injected into the tumor to observe the tumor volume and survival of mice. The changes in extracellular matrix were observed by Masson’s trichrome staining. Virus expression in tumor tissues was detected by immunofluorescence assay. The virus titer in tumor tissues was detected by TCID50. Histopathological changes were detected by hematoxylin and eosin(HE) and terminal deoxynucleotidyl transferase d UTP nick end labeling(TUNEL) staining. Intratumoral administration of NDV-MMP8 can effectively degrade ECM, promote the spread of the virus within the tumor, and reduce tumor growth rate. Therefore, the method of increasing intratumoral virus accumulation by degradation of the ECM to enhance the oncolytic effect has great potential for clinical application.
文摘Objective To study the clinical role of the variation of serum matrix metalloproteinase-8 (MMP-8) concentration in patients with acute coronary syndrome (ACS). Methods ELISA method was adopted to detect serum MMP-8 concentration and to observe concentration’s differences and features among 80 selected ACS cases (43 acute myocardial infarction and 37 unstable angina pectoris), 43 stable angina pectoris (SAP) cases and 37 control cases. And meanwhile the atherosclerosis risk factors of each case, such as age, sex, hypertension, body mass index, smoking, family history, diabetes, and hyperlipidemia were collected and analyzed as a whole. Results First, serum MMP-8 concentration reached the highest point in ACS, and there was significant difference between SAP and control groups (P<0.01). Second, serum MMP-8 in AMI was much higher than that in UAP with significant difference (P<0.01). There was no difference between UAP and SAP groups (P>0.05). Third, Logistic regression analysis revealed that serum MMP-8 concentration might be the indicator of ACS (B=4.493, P=0.000), particularly, that of AMI (B=9.961, P=0.000). Fourth, linear correlation and linear regression analysis found that only neutrophil was likely to influence serum MMP-8 concentration (r=0.274, P=0.001). Fifth, in the diagnosis of ACS, the area under ROC curve of MMP-8 was 0.785, the sensitivity and specificity were 68.6% and 76.5%, respectively. Conclusion ① Serum MMP-8 concentration has close relationship with the occurrence of ACS, particularly with AMI; ② Serum MMP-8 concentration may be one of the predicting indicators of ACS and particularly of AMI; ③ Neutrophil may be correlated with serum MMP-8 concentration; ④ MMP-8 is of somewhat valuable in diagnosing ACS.
文摘Background Sepsis-induced myocardial injury （SIMI） is caused by a variety of mechanisms. The aim of the study is to investigate the effects of metalloproteinase-8 （MMP-8） on SIMI and its mechanisms in rats. Methods Forty male Sprague Dawley rats were randomly divided into four groups： MMP-8 inhibitor （M81）, dexamethasone （DEX）, sepsis, and sham groups. The sepsis model was established by cecal ligation and puncture （CLP）. Rats in the M81 group immediately received an intraperitoneal injection of M81 （0.1 mg/kg） after CLP. Rats in the DEX group immediately received an intraperitoneal （IP） injection of DEX （2 mg/kg）. Rats in the sepsis and sham groups received intraperitoneal injections of normal saline. Rats were sacrificed 12 hours after CLP. Paraffin sections were stained with hematoxylin and eosin to observe the myocardium. The myocardial ultrastructure was observed with transmission electron microscopy. MMP-8, tumor necrosis factor-Q （TNF-a）, and interleukin-113 （IL-113） were detected by immunohistochemistry. The expression of MMP-8 was measured by Western blotting. TNF-a and IL-113 levels in serum and myocardial tissue were determined by enzyme-linked immunosorbent assay. Results Compared with the sham group, the myocardium in the sepsis group was seriously injured. MMP-8, TNF-α and IL-1β expression was higher in the sepsis group than in the sham group, Treatment with M81 or DEX, however, attenuated sepsis induced histopathological changes in the heart, and was associated with significant reductions in serum and myocardial levels of TNF-a and IL-113 （P 〈0.05）. M81 significantly inhibited MMP-8 expression in myocardial tissue （P 〈0.05）. In addition, treatment with DEX was not associated with a change in myocardial levels of MMP-8 （P 〉0.05）. Conclusion MMP-8 inhibitor attenuated myocardial injury in septic rats, which might be related to reduced expression of TNF-α and IL-1β.
文摘Periodontitis is an inflammatory disease of the periodontium. Any imbalance between the matrix metalloproteinases （MMPs） secreted by neutrophils and tissue inhibitors initiates the destruction of collagen in gum tissue, leading to chronic periodontitis. This study aimed to correlate salivary levels of MMP-8 and periodontal parameters of chronic periodontitis to establish MMP-8 as a noninvasive marker for the early diagnosis of chronic periodontitis. The study involved 40 subjects visiting the periodontic OPD of Dr. Ziauddin Ahmad Dental College and Hospital, located in Aligarh, U.P., India, from 2011 to 2012. The subjects were divided into two groups： group I consisted of 20 periodontally healthy subjects （controls） while group II consisted of 20 patients with chronic periodontitis. Chronic periodontitis was assessed on the basis of several periodontal parameters, including pocket probing depth （PPD）, clinical attachment level （CAL）, gingival index （GI）, and plaque index （PI）. Around 3 ml of unstimulated and whole expectorated saliva was collected for MMP-8 estimation by ELISA using Quantikine human total MMP-8 immunoassay kits. Data were analyzed using STATISTICA （Windows version 6） software. Salivary MMP-8 levels of groups I and II were 190.91 ± 143.89 ng/ml and 348.26± 202.1 ng/ml, respectively. The MMP-8 levels and periodontal status （PPD, CAL, GI, and PI） of groups I and II showed positive and significant correlations （for PPD, r = 0.63, P 〈 0.001; for CAL, r = 0.54, P 〈 0.001; for GI, r = 0.49, P 〈 0.001; and for PI, r = 0.63, P 〈 0.001）. The results of this study demonstrate elevated concentrations of MMP-8 in individuals with chronic periodontitis.
基金ThisworkwassupportedbyagrantfromtheNationalNaturalScienceFoundationofChina (No .3 0 2 0 0 2 5 4)
文摘Background Nasopharyngeal carcinoma (NPC) shows highly invasive and metastatic features. This study aims to investigate macrophage migration inhibitory factor (MIF)-induced invasion of NPC cells in vitro and the effects on matrix metalloproteinases (MMPs) and interleukin-8 (IL-8),and to study the mechanism of tumor cell invasion and metastasis in the early stage of NPC. Methods Two nasopharyngeal carcinoma cell lines,CNE-1 and CNE-2,were adopted in this study. The NPC cell invasion and migration were evaluated by microinvasion assay. The variation of expression percentages of MMP2- or MMP9-positive cells was detected by flow cytometry in two cell lines with or without MIF treatment. Western blotting and RT-PCR were used to assay the protein and mRNA expressions of MMP2 and MMP9. The IL-8 concentration secreted by NPC cells was compared with the cells with different treatments using ELISA. Results After treating with MIF for 48 hours,the cell numbers of CNE-1 and CNE-2 which went through the 8-μm filter membrane were increased. Compared with non-MIF treated NPC cells,significant difference could be found both in CNE-1( P =0.005) and CNE-2 cells ( P =0.001) . The percentages of MMP9-positive cells were significantly increased in both CNE-1 [from (28.5±2.5)% to (82.4±3.5)%, P =0.001] and CNE-2 [from (32.8±3.5)% to (86.1±1.6)%, P =0.002]. The relative intensity of MMP9 protein expression was also enhanced in both cell lines (CNE-1: from 83.1±6.0 to 242.9±22.9, P =0.002;CNE-2: from 84.4±4.3 to 278.9±29.7, P =0.003). Correspondingly,the increased MMP9 mRNA expression level was significantly detectable in both cell lines.The concentration of IL-8 in the supernatant of CNE-2 was higher [(1201.8±593.3) pg/ml] after treatment. It was also remarkably higher than that in the supernatant of CNE-2 without treatment ( P =0.026). However,there was no significant difference in the concentration variation of IL-8 in CNE-1 ( P =0.581), while the IL-8 mRNA level was only enhanced in CNE-2. Conclusions MIF can induce potent invasion of NPC cell lines in vitro , and the infiltrating lymphocytes in NPC might be responsible for the invasion and metastasis of tumor cells. MIF cytokine which is secreted by these infiltrating lymphocytes might contribute to the invasion as well as metastasis of NPC in the early stages by induction of MMP9 and IL-8 in an indirect pathway.
