Signals of touch,pressure,pain,temperature,position,and vibration are transmitted from the peripheral nervous system（PNS）to the dorsal horn of the segmental spinal cord via pseudounipolar dorsal root ganglia（DRG）n...Signals of touch,pressure,pain,temperature,position,and vibration are transmitted from the peripheral nervous system（PNS）to the dorsal horn of the segmental spinal cord via pseudounipolar dorsal root ganglia（DRG）neurons.Sensory information gathering relies on functional integrity of DRG neurons and can be interrupted by PNS injury due to trauma,disease or exposure to drugs, toxins or viral pathogens. Despite the high regenerative capacity of DRG neurons, sensory recovery after PNS injury is often incomplete and severe neuropathic pain may last for years. Although numerous mechanisms of PNS injury have been established, neuropathic pain is refractory to therapy, contributing to the physical and emotional suffering of patients and the enormous economic burden to society.展开更多
In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 (TIMP-2) from Japanese puffer fishFugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2...In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 (TIMP-2) from Japanese puffer fishFugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2b cDNAsare composed of 660 and 657 nucleotides and 220 and 219 amino acids, respectively. Both Fugu TIMP-2s contain 12 cysteineresidues, which might form six disulfide bonds as in other animals’ TIMP-2s. Reverse-transcribed polymerase chain reactionanalysis showed the mRNAs of Fugu TIMP-2a and TIMP-2b to be expressed in some tissues examined with different expres-sion patterns. These findings suggest that the two distinct Fugu TIMP-2s might perform different functions in Fugu tissues.展开更多
To investigate the effects of TGF-β1 on the two gelatinases （MMP-2 and MMP-9）, and their roles in lung remodeling after irradiation-induced lung injury. Expressions of TGF-β1 were measured with western blot, and e...To investigate the effects of TGF-β1 on the two gelatinases （MMP-2 and MMP-9）, and their roles in lung remodeling after irradiation-induced lung injury. Expressions of TGF-β1 were measured with western blot, and expressions of MMP-2 and MMP-9 were analyzed with zymography in a TGF-β1 transgenic mouse model after thoracic irradiation with 12 Gy. We found expressions of TGF-β1 in the lung from the transgenic mice were three folds as compared to those from control mice. With densitometrical analysis, we found a significant decrease in MMP-9 activity in lung homogenates from the transgenic mice as compared with those from non-transgenic control mice 8 weeks after sham-irradiation （relative MMP-9 activity： C： 1. 000±0. 1091; TG： 0. 4772±0. 470 （n=8, P〈0.05）. But MMP-2 was constitutively expressed in the lung homogenates from the transgenic mice as compared to those from control mice 8 weeks after sham-irradiation （relative MMP-2 activity 8 weeks after sham-irradiation： C： 1. 000±0. 1556, TG： 1. 0075±0. 1472）. Eight weeks after thoracic irradiation with 12 Gy, we observed a significant increase of MMP-2 and MMP-9 activity in lung homogenates from both transgenic and normal mice. In TGF-β1 transgenic mice relative MMP-9 activity was increased to 1. 5321±0. 2217 folds 8 weeks after thoracic irradiation with 12 Gy as compared to those after sham-irradiation （1. 000±0. 2153）, and relative MMP-2 activity was increased to 1. 7142 ± 0. 4231 folds. Our results show that TGF-β1 itself down-regulates activity of MMP-9, thereby decreases ECM degradation in lungs of TGF-β1 transgenic mice. Also we find that ionizing irradiation upregulates both MMP-2 and MMP-9 activity. Over-expressions of MMP-9 and MMP-2 after lung irradiation are involved in the inflammatory response associated with radiation-induced lung injury, and maybe further in radiation-induced lung fibrosis.展开更多
This study aimed to explore the molecular mechanism in tumor invasion and metastasis. The ex-pression of matrix metalloproteinase-2,-9 (MMP-2,MMP-9), tissue inhibitor-1 of matrix metalloprote inase(TIMP-1) , cell adh...This study aimed to explore the molecular mechanism in tumor invasion and metastasis. The ex-pression of matrix metalloproteinase-2,-9 (MMP-2,MMP-9), tissue inhibitor-1 of matrix metalloprote inase(TIMP-1) , cell adhesion molecule 44 variant 6 (CD44v6) , HER2/neu and p53 was investigated in 154 pa-tients with head and neck squamous cell carcinoma (SCC) by ABC and ImmunoMax immunohistochemical method. Their clinical relevance and correlation were analysed. The expression of MMP-2, MMP-9, TIMP-1,CD44v6, HER2/neu and p53 was found in cancer cells in 87.01%, 85.71%, 68. 18%, 98.05%,55.19% and 50.65% cases respectively. Linear regression and correlation analysis revealed that there wasclose positive relationship ( P < 0.05) between the expression of MMP-2 and MMP-9, TIMP-1 and CD44v6,HER2/neu and MMP-9, MMP-2 and p53. Up-regulation of MMP-2 was accompanied by advanced T stage( P < 0.01 ) . There was also a trend of MMP-2 expression being related with tumor metastasis. Increased ex-pression of HER2/neu was found in patients with tumor recurrence( P < 0.05 ) . The expression of TIMP-1 washigher in laryngeal cancer than that in pharyngeal cancer, and higher in keratinizing and non-keratlnizing SCC than that in basaloid SCC ( P < 0.05 ) . These findings suggested that MMP-2 and MMP-9, HER2/neu andMMP-9, MMP-2 and p53 had a coordinate function in aggression of tumor; that MMP-2 had a more important function than MMP-9 in tumor invasion and metastasis; and that HER2/neu might serve as a biomarker forpoor prognosis in HNSCC.展开更多
目的探讨AKI risk评分(基质金属蛋白酶-2×胰岛素样生长因子-7,TIMP-2×IGFBP-7)对急诊脓毒症患者死亡风险的预测价值。方法前瞻性观察2021年9月至2022年12月中国科学技术大学附属第一医院及北京协和医院急诊科收住的脓毒症患者...目的探讨AKI risk评分(基质金属蛋白酶-2×胰岛素样生长因子-7,TIMP-2×IGFBP-7)对急诊脓毒症患者死亡风险的预测价值。方法前瞻性观察2021年9月至2022年12月中国科学技术大学附属第一医院及北京协和医院急诊科收住的脓毒症患者,分别测量患者入院时和入院后6 h的AKI risk评分并计算其变化值(AKI risk-gap),利用多因素Logistic回归、Cox回归、受试者工作特征(ROC)曲线及曲线下面积(AUC)分析AKI risk评分对患者院内死亡风险的预测效能;亚组分析中根据患者是否罹患AKI进一步分析AKI risk评分与不同亚组(AKI组和非AKI组)患者预后的关系。结果本研究共纳入患者202例,住院期间死亡87例(43%)。ROC曲线显示,6 h AKI risk评分预测脓毒症患者院内死亡最为准确,其AUC为0.71(95%CI 0.63~0.78)。亚组分析中AKI组患者6 h AKI risk评分预测院内死亡的AUC为0.76(95%CI 0.65~0.85),非AKI组AUC为0.63(95%CI 0.52~0.73)。多因素Logistic回归和Cox回归分析表明,6 h AKI risk评分和AKI risk-gap是患者院内死亡的独立危险因素。结论AKI risk评分对脓毒症患者院内死亡风险有较好的预测价值,尤其6 h AKI risk评分在罹患AKI的亚组患者中预测价值最高,可为临床区分高危患者并给予相应治疗提供参考。展开更多
目的全面分析乙型肝炎病毒X蛋白(Hepatitis B virus X protein,HBx)对基质金属蛋白酶(Matrix Metallo-proteinases,MMPs)及组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases,TIMPs)的影响,探讨其在肝细胞癌的侵袭转移中...目的全面分析乙型肝炎病毒X蛋白(Hepatitis B virus X protein,HBx)对基质金属蛋白酶(Matrix Metallo-proteinases,MMPs)及组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases,TIMPs)的影响,探讨其在肝细胞癌的侵袭转移中的可能作用。方法PCR扩增HBVX基因并克隆入真核表达载体pcDNA3.1/HisC,重组载体及空载体分别以Lipofectamine2000转染HepG2细胞并以800μg/mlG418筛选抗性细胞克隆。以Western blot检测抗性细胞HBx表达。抽提细胞总RNA,半定量RT-PCR检测MMPs及TIMPs。收集细胞培养液上清,以明胶酶谱检测MMP2及MMP9活性,反相明胶酶谱检测TIMPs活性。结果构建了HBx重组载体pcDNA3.1-XB。该重组载体及对照空载体转染HepG2细胞后,G418筛选,分别获得抗性细胞克隆HepG2-XB及HepG2-HIS,前者经Western blot证实可表达HBx。半定量RT-PCR显示HBx可促进MMP2、7、13、14、16、17、19、23、24及TIMP1、4基因的转录,抑制MMP1、3、8、9、10、11、12、15、20及TIMP2、3基因的转录。明胶酶谱检测显示HBx可促进酶原MMP2(Pro-MMP2)及活性MMP9(Active-MMP9)表达,抑制酶原MMP9(Pro-MMP9)表达;反相明胶酶谱显示HBx可促进TIMP1、TIMP4的表达,同时抑制TIMP2及糖基化TIMP3的表达。结论HBx蛋白对MMPs、TIMPs转录表达的影响是多方面的,但这种影响在HBx促进肝癌细胞侵袭转移过程中的确切机制有待进一步阐明。展开更多
Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometri...Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometrial stromal cells separated from the proliferative endometrial tissues were incubated with medium alone, 17-β estradiol （E2,10^-8 mol/L）, medroxyprogesterone acetate （MPA, 10^-6 mol/L）, E2（10^-8 mol/L）＋MPA （10^-6 mol/L）, E2 （10^-8 mol/L）＋MPA （10^-6 mol/L）＋RU486 （10^-5 mol/L） or HB-EGF （10 ng/ml） for 48 h respectively. The expressions of MMP-9 and TIMP-1 were detected by in situ hybridization, immunocytochemistry, reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting. Results Compared with control group [mRNA, 0. 729 ± 0. 090 （MMP-9） and 1.056± 0.154 （TIMP-1）; protein, 0.545 ±0.086 （MMP-9） and 0.745 ±0.154 （TIMP-1）], expressions of MMP-9 and TIMP-1 in E2 alone, progestin alone or E2 combined with progestin group were respectively：mRNA, 0.413 ± 0.069, 0.402 ± 0.073 and 0.407 ± 0.039; 0.487 ± 0.093, 0.503 ± 0.093 and 0.468 ± 0.075：protein, 0.294 ± 0.076, 0.331 ±0.064 and 0.265 ±0.049; 0.425 ±0.085, 0.397 ±0.065 and 0.435 ± 0.099. RU486 weakened the expression level of down-regulation, while HB-EGF elevated the level of MMP-9 and TIMP-1 after 48 h treatment （mRNA, 0.955 ± 0.068 and 1.396 ± 0.238; protein, 0. 780 ± 0.109 and 0.985 ± 0.165）. Conclusions 1） Both E2 and progestin can down-regulate the expressions of MMP-9 and TIMP-1 in endometrial stromal cells, but RU486 can inhibit the effect. 2） HB-EGF can elevate the level of MMP-9 and TIMP-1. 3） E2, progestin and HB-EGF have effect on the ratio of MM-P/TIMP-1.展开更多
基金Department of Veteran’s Affairs Merit Review Award 5I01BX000638(to VIS)National Institutes of Health R01DE022757(to VIS and AYS)supported this study
文摘Signals of touch,pressure,pain,temperature,position,and vibration are transmitted from the peripheral nervous system（PNS）to the dorsal horn of the segmental spinal cord via pseudounipolar dorsal root ganglia（DRG）neurons.Sensory information gathering relies on functional integrity of DRG neurons and can be interrupted by PNS injury due to trauma,disease or exposure to drugs, toxins or viral pathogens. Despite the high regenerative capacity of DRG neurons, sensory recovery after PNS injury is often incomplete and severe neuropathic pain may last for years. Although numerous mechanisms of PNS injury have been established, neuropathic pain is refractory to therapy, contributing to the physical and emotional suffering of patients and the enormous economic burden to society.
基金partially supported by a grant from the Ministry of Education,Culture,Sports,Science and Technology of Japan.
文摘In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 (TIMP-2) from Japanese puffer fishFugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2b cDNAsare composed of 660 and 657 nucleotides and 220 and 219 amino acids, respectively. Both Fugu TIMP-2s contain 12 cysteineresidues, which might form six disulfide bonds as in other animals’ TIMP-2s. Reverse-transcribed polymerase chain reactionanalysis showed the mRNAs of Fugu TIMP-2a and TIMP-2b to be expressed in some tissues examined with different expres-sion patterns. These findings suggest that the two distinct Fugu TIMP-2s might perform different functions in Fugu tissues.
文摘To investigate the effects of TGF-β1 on the two gelatinases （MMP-2 and MMP-9）, and their roles in lung remodeling after irradiation-induced lung injury. Expressions of TGF-β1 were measured with western blot, and expressions of MMP-2 and MMP-9 were analyzed with zymography in a TGF-β1 transgenic mouse model after thoracic irradiation with 12 Gy. We found expressions of TGF-β1 in the lung from the transgenic mice were three folds as compared to those from control mice. With densitometrical analysis, we found a significant decrease in MMP-9 activity in lung homogenates from the transgenic mice as compared with those from non-transgenic control mice 8 weeks after sham-irradiation （relative MMP-9 activity： C： 1. 000±0. 1091; TG： 0. 4772±0. 470 （n=8, P〈0.05）. But MMP-2 was constitutively expressed in the lung homogenates from the transgenic mice as compared to those from control mice 8 weeks after sham-irradiation （relative MMP-2 activity 8 weeks after sham-irradiation： C： 1. 000±0. 1556, TG： 1. 0075±0. 1472）. Eight weeks after thoracic irradiation with 12 Gy, we observed a significant increase of MMP-2 and MMP-9 activity in lung homogenates from both transgenic and normal mice. In TGF-β1 transgenic mice relative MMP-9 activity was increased to 1. 5321±0. 2217 folds 8 weeks after thoracic irradiation with 12 Gy as compared to those after sham-irradiation （1. 000±0. 2153）, and relative MMP-2 activity was increased to 1. 7142 ± 0. 4231 folds. Our results show that TGF-β1 itself down-regulates activity of MMP-9, thereby decreases ECM degradation in lungs of TGF-β1 transgenic mice. Also we find that ionizing irradiation upregulates both MMP-2 and MMP-9 activity. Over-expressions of MMP-9 and MMP-2 after lung irradiation are involved in the inflammatory response associated with radiation-induced lung injury, and maybe further in radiation-induced lung fibrosis.
文摘This study aimed to explore the molecular mechanism in tumor invasion and metastasis. The ex-pression of matrix metalloproteinase-2,-9 (MMP-2,MMP-9), tissue inhibitor-1 of matrix metalloprote inase(TIMP-1) , cell adhesion molecule 44 variant 6 (CD44v6) , HER2/neu and p53 was investigated in 154 pa-tients with head and neck squamous cell carcinoma (SCC) by ABC and ImmunoMax immunohistochemical method. Their clinical relevance and correlation were analysed. The expression of MMP-2, MMP-9, TIMP-1,CD44v6, HER2/neu and p53 was found in cancer cells in 87.01%, 85.71%, 68. 18%, 98.05%,55.19% and 50.65% cases respectively. Linear regression and correlation analysis revealed that there wasclose positive relationship ( P < 0.05) between the expression of MMP-2 and MMP-9, TIMP-1 and CD44v6,HER2/neu and MMP-9, MMP-2 and p53. Up-regulation of MMP-2 was accompanied by advanced T stage( P < 0.01 ) . There was also a trend of MMP-2 expression being related with tumor metastasis. Increased ex-pression of HER2/neu was found in patients with tumor recurrence( P < 0.05 ) . The expression of TIMP-1 washigher in laryngeal cancer than that in pharyngeal cancer, and higher in keratinizing and non-keratlnizing SCC than that in basaloid SCC ( P < 0.05 ) . These findings suggested that MMP-2 and MMP-9, HER2/neu andMMP-9, MMP-2 and p53 had a coordinate function in aggression of tumor; that MMP-2 had a more important function than MMP-9 in tumor invasion and metastasis; and that HER2/neu might serve as a biomarker forpoor prognosis in HNSCC.
文摘目的探讨AKI risk评分(基质金属蛋白酶-2×胰岛素样生长因子-7,TIMP-2×IGFBP-7)对急诊脓毒症患者死亡风险的预测价值。方法前瞻性观察2021年9月至2022年12月中国科学技术大学附属第一医院及北京协和医院急诊科收住的脓毒症患者,分别测量患者入院时和入院后6 h的AKI risk评分并计算其变化值(AKI risk-gap),利用多因素Logistic回归、Cox回归、受试者工作特征(ROC)曲线及曲线下面积(AUC)分析AKI risk评分对患者院内死亡风险的预测效能;亚组分析中根据患者是否罹患AKI进一步分析AKI risk评分与不同亚组(AKI组和非AKI组)患者预后的关系。结果本研究共纳入患者202例,住院期间死亡87例(43%)。ROC曲线显示,6 h AKI risk评分预测脓毒症患者院内死亡最为准确,其AUC为0.71(95%CI 0.63~0.78)。亚组分析中AKI组患者6 h AKI risk评分预测院内死亡的AUC为0.76(95%CI 0.65~0.85),非AKI组AUC为0.63(95%CI 0.52~0.73)。多因素Logistic回归和Cox回归分析表明,6 h AKI risk评分和AKI risk-gap是患者院内死亡的独立危险因素。结论AKI risk评分对脓毒症患者院内死亡风险有较好的预测价值,尤其6 h AKI risk评分在罹患AKI的亚组患者中预测价值最高,可为临床区分高危患者并给予相应治疗提供参考。
文摘目的全面分析乙型肝炎病毒X蛋白(Hepatitis B virus X protein,HBx)对基质金属蛋白酶(Matrix Metallo-proteinases,MMPs)及组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases,TIMPs)的影响,探讨其在肝细胞癌的侵袭转移中的可能作用。方法PCR扩增HBVX基因并克隆入真核表达载体pcDNA3.1/HisC,重组载体及空载体分别以Lipofectamine2000转染HepG2细胞并以800μg/mlG418筛选抗性细胞克隆。以Western blot检测抗性细胞HBx表达。抽提细胞总RNA,半定量RT-PCR检测MMPs及TIMPs。收集细胞培养液上清,以明胶酶谱检测MMP2及MMP9活性,反相明胶酶谱检测TIMPs活性。结果构建了HBx重组载体pcDNA3.1-XB。该重组载体及对照空载体转染HepG2细胞后,G418筛选,分别获得抗性细胞克隆HepG2-XB及HepG2-HIS,前者经Western blot证实可表达HBx。半定量RT-PCR显示HBx可促进MMP2、7、13、14、16、17、19、23、24及TIMP1、4基因的转录,抑制MMP1、3、8、9、10、11、12、15、20及TIMP2、3基因的转录。明胶酶谱检测显示HBx可促进酶原MMP2(Pro-MMP2)及活性MMP9(Active-MMP9)表达,抑制酶原MMP9(Pro-MMP9)表达;反相明胶酶谱显示HBx可促进TIMP1、TIMP4的表达,同时抑制TIMP2及糖基化TIMP3的表达。结论HBx蛋白对MMPs、TIMPs转录表达的影响是多方面的,但这种影响在HBx促进肝癌细胞侵袭转移过程中的确切机制有待进一步阐明。
文摘Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometrial stromal cells separated from the proliferative endometrial tissues were incubated with medium alone, 17-β estradiol （E2,10^-8 mol/L）, medroxyprogesterone acetate （MPA, 10^-6 mol/L）, E2（10^-8 mol/L）＋MPA （10^-6 mol/L）, E2 （10^-8 mol/L）＋MPA （10^-6 mol/L）＋RU486 （10^-5 mol/L） or HB-EGF （10 ng/ml） for 48 h respectively. The expressions of MMP-9 and TIMP-1 were detected by in situ hybridization, immunocytochemistry, reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting. Results Compared with control group [mRNA, 0. 729 ± 0. 090 （MMP-9） and 1.056± 0.154 （TIMP-1）; protein, 0.545 ±0.086 （MMP-9） and 0.745 ±0.154 （TIMP-1）], expressions of MMP-9 and TIMP-1 in E2 alone, progestin alone or E2 combined with progestin group were respectively：mRNA, 0.413 ± 0.069, 0.402 ± 0.073 and 0.407 ± 0.039; 0.487 ± 0.093, 0.503 ± 0.093 and 0.468 ± 0.075：protein, 0.294 ± 0.076, 0.331 ±0.064 and 0.265 ±0.049; 0.425 ±0.085, 0.397 ±0.065 and 0.435 ± 0.099. RU486 weakened the expression level of down-regulation, while HB-EGF elevated the level of MMP-9 and TIMP-1 after 48 h treatment （mRNA, 0.955 ± 0.068 and 1.396 ± 0.238; protein, 0. 780 ± 0.109 and 0.985 ± 0.165）. Conclusions 1） Both E2 and progestin can down-regulate the expressions of MMP-9 and TIMP-1 in endometrial stromal cells, but RU486 can inhibit the effect. 2） HB-EGF can elevate the level of MMP-9 and TIMP-1. 3） E2, progestin and HB-EGF have effect on the ratio of MM-P/TIMP-1.
