Background Previous studies have indicated that thrombi n (TM) may play a major role in brain edema after intracerebral hemorrhages (ICHs). However, the mechanism of TM-induced brain edema is poorly understood. In th...Background Previous studies have indicated that thrombi n (TM) may play a major role in brain edema after intracerebral hemorrhages (ICHs). However, the mechanism of TM-induced brain edema is poorly understood. In this study, we explored the effect of TM on the permeability of the blood brain barrier (BBB) and investigated its possible mechanism, aiming at providing a potential target for brain edema therapy after ICHs.Methods TM or TM + cathepsin G (CATG) was stereotaxically injected into the right caudate nucleus of Sprague-Dawley rats in vivo. BBB permeability was measured by Evans-Blue extravasation. Brain water content was determined by the dry-wet weight method. Brain microvascular endothelial cells were then cultured in vitro. After TM or TM+CATG was added to the endothelial cell medium, changes in the morphology of cells were dynamically observed by phase-contrast light microscopy, and the expression of matrix metalloproteinase-2 (MMP-2) protein was measured by immunohistochemical method.Results BBB permeability increased at 6 hours after a TM injection into the ipsilateral caudate nucleus (P<0.05), peaked between 24 hours (P<0.01) and 48 hours (P<0.05) after the injection, and then declined. Brain water content changed in parallel with the changes in BBB permeability. However, at all time points, BBB permeability and brain water content after a TM+CATG injection were not significantly different from the respective parameters in the control group (P>0.05). TM induced endothelial cell contraction in vitro in a time-dependent manner and enhanced the expression of MMP-2 protein. After incubation with TM+CATG, cell morphology and MMP-2 expression did not change significantly as compared to the control group (P>0.05).Conclusions Increased BBB permeability may be one of the mechanisms behind TM-induced cerebral edema. TM induces endothelial cell contraction and promotes MMP-2 expression by activating protease activated receptor-1 (PAR-1), possibly leading to the opening of the BBB.展开更多
Background Cathepsin S and its endogenous inhibitor cystatin C are implicated in the pathogenesis of atherosclerosis, especially in the plaque destabilization and rupture leading to acute coronary syndrome. However, w...Background Cathepsin S and its endogenous inhibitor cystatin C are implicated in the pathogenesis of atherosclerosis, especially in the plaque destabilization and rupture leading to acute coronary syndrome. However, whether circulating cathepsin S and cystatin C also change in association with coronary plaque morphology is unknown yet. Methods We recruited 98 patients with unstable angina （UA, n=56） or stable angina （SA, n=-42） who had a segmental stenosis resulting in 〉20% and 〈70% diameter reduction in one major coronary artery on coronary angiography. Thirty-one healthy subjects served as controls. Intravascular ultrasound （IVUS） was used to evaluate plaque morphology. Plasma cathepsin S and cystatin C were measured as well. Results At the culprit lesion site, plaque area （（7.85±2.83） mm^2 vs （6.53±2.92） mm^2, P=0.027）, plaque burden （（60.92±11.04）% vs （53.87±17.52）%, P=0.025）, remodeling index （0.93±0.16 vs 0.86±0.10, P=0.004） and eccentricity index （0.74±0.17 vs 0.66±0.21, P=0.038） were bigger in UA group than in SA group. Plasma cathepsin S and cystatin C were significantly higher in patients than in controls （P〈0.01）. Plasma cathepsin S was higher in UA group （（0.411±0.121） nmol/L） than in SA group （（0.355±0.099） nmol/L, P=0.007）, so did the plasma cystatin C （（0.95±0.23） mg/L in UA group, （0.84±0.22） mg/L in SA group; P=0.009）. Plasma cathepsin S positively correlated with remodeling index （r=0.402, P=0.002） and eccentricity index （r=0.441, P=0.001）, and plasma cystatin C positively correlated with plaque area （r=0.467, P 〈0.001） and plaque burden （r=0.395, P=0.003） in UA group but not in SA group. Conclusions Plasma cathepsin S and cystatin C increased significantly in UA patients. In angina patients, higher plasma cathepsin S may suggest the presence of vulnerable plaque, and higher plasma cystatin C may be a clue for larger atherosclerotic coronary plaque.展开更多
The changes of tissue appearances and structures in the process of UV-induced ＂melting＂ for sea cucumber （Stichopus japonicus） body wall were studied. And the localization and determination of acid phosphatase （...The changes of tissue appearances and structures in the process of UV-induced ＂melting＂ for sea cucumber （Stichopus japonicus） body wall were studied. And the localization and determination of acid phosphatase （ACP）, Cathepsin B and Cathepsin L activities were also investigated. The results show that the connective tissue was damaged with many hollows emerging and the regular collagen bundles were broken apart into irregular fragments. Margination of condensed chromatin at the nuclear membrane was observed. Both Golgi＇s body and endoplasmic reticulure swelled, curled, and eventually double- or multi-lamellar vesicles were formed. A number of autophagic vesicles distributed in all through the whole cytoplasm. ACP becomes more active after UV irradiation. The activities of cathepsin B and cathepsin L increased in UV-treated sea cucumbers and both achieved their maximum under certain conditions, It indicates that autophagy plays a potential role in the ＂melting＂ process for sea cucumber body wall induced by UV irradiation.展开更多
It remains unclear whether autophagy affects hippocampal neuronal injury in vascular dementia. In the present study, we investigated the effects of autophagy blockade on hippocampal neuro- nal injury in a rat model of...It remains unclear whether autophagy affects hippocampal neuronal injury in vascular dementia. In the present study, we investigated the effects of autophagy blockade on hippocampal neuro- nal injury in a rat model of vascular dementia. In model rats, hippocampal CA1 neurons were severely damaged, and expression of the autophagy-related proteins beclin-1, cathepsin B and microtubule-associated protein 1 light chain 3 was elevated compared with that in sham-operated animals. These responses were suppressed in animals that received a single intraperitoneal injection of wortmannin, an autophagy inhibitor, prior to model establishment. The present results confirm that autophagy and autophagy-related proteins are involved in the pathological changes of vascular dementia, and that inhibition of autophagy has neuroprotective effects.展开更多
Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment fro...Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing. Cathepsin K was expressed in a high amount in E.coli after IPTG induction, and was purified to near homogenetity through resolution and column purification. The specificity of the protein was shown by Western blotting experiment. The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain. Results With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects. The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K. Conclusions The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization.展开更多
Cystatin C,cathepsin S,and IL-1 are three important biomarkers of atherosclerosis.Previous studies emphasized the relationship between individual biomarkers in coronary artery disease（CAD） patients and severity of a...Cystatin C,cathepsin S,and IL-1 are three important biomarkers of atherosclerosis.Previous studies emphasized the relationship between individual biomarkers in coronary artery disease（CAD） patients and severity of atherosclerostic lesions of the coronary arteries,while combined cystatin C,cathepsin S,and IL-1 have not been reported for clinical classification of CAD.We aimed to establish a link between cystatin C,cathepsin S,IL-1 and CAD in this cohort study.Totally 112 subjects were enrolled and divided into the stable angina pectoris group,the unstable angina pectoris group and the acute myocardial infarction（AMI） groups,and 50 healthy adults served as controls.The levels of the three biomarkers were detected by ELISA.The results showed that serum level of cystatin C（mg/L） was higher in CAD patients compared with those in the healthy controls（AMI vs.unstable angina pectoris vs.stable angina pectoris vs.controls：1.27±0.18 vs.1.09±0.19 vs.0.91±0.05 vs.0.78±0.07,all P〈0.01）.Cathepsin S（ng/mL） was also significantly different among the groups（AMI vs.unstable angina pectoris vs.stable angina pectoris vs.controls：67.30±8.36 vs.56.90±7.16 vs.49.8±2.72 vs.67.30±8.36,all P〈0.01）.IL-1（pg/mL） was significantly different among the groups as well（AMI vs.unstable angina pectoris vs.stable angina pectoris vs.controls：2.96±0.57 vs.2.46±0.24 vs.2.28±0.09 vs.2.02±0.13,all P〈0.01）.Spearman＇s correlation test revealed positive correlation between cystatin C,cathepsin S,IL-1 and Gensini score（r=0.451,0.491,0.397,respectively）.It is suggested that simultaneous detection of cystatin C,cathepsin S,and IL-1 in serum may be useful in clinical classification and assessment of severity of CAD.展开更多
文摘Background Previous studies have indicated that thrombi n (TM) may play a major role in brain edema after intracerebral hemorrhages (ICHs). However, the mechanism of TM-induced brain edema is poorly understood. In this study, we explored the effect of TM on the permeability of the blood brain barrier (BBB) and investigated its possible mechanism, aiming at providing a potential target for brain edema therapy after ICHs.Methods TM or TM + cathepsin G (CATG) was stereotaxically injected into the right caudate nucleus of Sprague-Dawley rats in vivo. BBB permeability was measured by Evans-Blue extravasation. Brain water content was determined by the dry-wet weight method. Brain microvascular endothelial cells were then cultured in vitro. After TM or TM+CATG was added to the endothelial cell medium, changes in the morphology of cells were dynamically observed by phase-contrast light microscopy, and the expression of matrix metalloproteinase-2 (MMP-2) protein was measured by immunohistochemical method.Results BBB permeability increased at 6 hours after a TM injection into the ipsilateral caudate nucleus (P<0.05), peaked between 24 hours (P<0.01) and 48 hours (P<0.05) after the injection, and then declined. Brain water content changed in parallel with the changes in BBB permeability. However, at all time points, BBB permeability and brain water content after a TM+CATG injection were not significantly different from the respective parameters in the control group (P>0.05). TM induced endothelial cell contraction in vitro in a time-dependent manner and enhanced the expression of MMP-2 protein. After incubation with TM+CATG, cell morphology and MMP-2 expression did not change significantly as compared to the control group (P>0.05).Conclusions Increased BBB permeability may be one of the mechanisms behind TM-induced cerebral edema. TM induces endothelial cell contraction and promotes MMP-2 expression by activating protease activated receptor-1 (PAR-1), possibly leading to the opening of the BBB.
文摘Background Cathepsin S and its endogenous inhibitor cystatin C are implicated in the pathogenesis of atherosclerosis, especially in the plaque destabilization and rupture leading to acute coronary syndrome. However, whether circulating cathepsin S and cystatin C also change in association with coronary plaque morphology is unknown yet. Methods We recruited 98 patients with unstable angina （UA, n=56） or stable angina （SA, n=-42） who had a segmental stenosis resulting in 〉20% and 〈70% diameter reduction in one major coronary artery on coronary angiography. Thirty-one healthy subjects served as controls. Intravascular ultrasound （IVUS） was used to evaluate plaque morphology. Plasma cathepsin S and cystatin C were measured as well. Results At the culprit lesion site, plaque area （（7.85±2.83） mm^2 vs （6.53±2.92） mm^2, P=0.027）, plaque burden （（60.92±11.04）% vs （53.87±17.52）%, P=0.025）, remodeling index （0.93±0.16 vs 0.86±0.10, P=0.004） and eccentricity index （0.74±0.17 vs 0.66±0.21, P=0.038） were bigger in UA group than in SA group. Plasma cathepsin S and cystatin C were significantly higher in patients than in controls （P〈0.01）. Plasma cathepsin S was higher in UA group （（0.411±0.121） nmol/L） than in SA group （（0.355±0.099） nmol/L, P=0.007）, so did the plasma cystatin C （（0.95±0.23） mg/L in UA group, （0.84±0.22） mg/L in SA group; P=0.009）. Plasma cathepsin S positively correlated with remodeling index （r=0.402, P=0.002） and eccentricity index （r=0.441, P=0.001）, and plasma cystatin C positively correlated with plaque area （r=0.467, P 〈0.001） and plaque burden （r=0.395, P=0.003） in UA group but not in SA group. Conclusions Plasma cathepsin S and cystatin C increased significantly in UA patients. In angina patients, higher plasma cathepsin S may suggest the presence of vulnerable plaque, and higher plasma cystatin C may be a clue for larger atherosclerotic coronary plaque.
基金the National Natural Science Foundation of China(30571449)the National Basic Research Program of China (2006CB708210)the Chinese International Corporation Project (2006DFA32580)
文摘The changes of tissue appearances and structures in the process of UV-induced ＂melting＂ for sea cucumber （Stichopus japonicus） body wall were studied. And the localization and determination of acid phosphatase （ACP）, Cathepsin B and Cathepsin L activities were also investigated. The results show that the connective tissue was damaged with many hollows emerging and the regular collagen bundles were broken apart into irregular fragments. Margination of condensed chromatin at the nuclear membrane was observed. Both Golgi＇s body and endoplasmic reticulure swelled, curled, and eventually double- or multi-lamellar vesicles were formed. A number of autophagic vesicles distributed in all through the whole cytoplasm. ACP becomes more active after UV irradiation. The activities of cathepsin B and cathepsin L increased in UV-treated sea cucumbers and both achieved their maximum under certain conditions, It indicates that autophagy plays a potential role in the ＂melting＂ process for sea cucumber body wall induced by UV irradiation.
基金supported by the Scientific Technology Research Project of Hebei Provincial Higher Learning Schools in China,No.ZH2012046the Major Medical Research Program of Hebei Province in China,No.ZD2013087
文摘It remains unclear whether autophagy affects hippocampal neuronal injury in vascular dementia. In the present study, we investigated the effects of autophagy blockade on hippocampal neuro- nal injury in a rat model of vascular dementia. In model rats, hippocampal CA1 neurons were severely damaged, and expression of the autophagy-related proteins beclin-1, cathepsin B and microtubule-associated protein 1 light chain 3 was elevated compared with that in sham-operated animals. These responses were suppressed in animals that received a single intraperitoneal injection of wortmannin, an autophagy inhibitor, prior to model establishment. The present results confirm that autophagy and autophagy-related proteins are involved in the pathological changes of vascular dementia, and that inhibition of autophagy has neuroprotective effects.
文摘Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing. Cathepsin K was expressed in a high amount in E.coli after IPTG induction, and was purified to near homogenetity through resolution and column purification. The specificity of the protein was shown by Western blotting experiment. The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain. Results With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects. The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K. Conclusions The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization.
基金founded by science and technology planning project of Xuzhou City(No.KC14SH088)
文摘Cystatin C,cathepsin S,and IL-1 are three important biomarkers of atherosclerosis.Previous studies emphasized the relationship between individual biomarkers in coronary artery disease（CAD） patients and severity of atherosclerostic lesions of the coronary arteries,while combined cystatin C,cathepsin S,and IL-1 have not been reported for clinical classification of CAD.We aimed to establish a link between cystatin C,cathepsin S,IL-1 and CAD in this cohort study.Totally 112 subjects were enrolled and divided into the stable angina pectoris group,the unstable angina pectoris group and the acute myocardial infarction（AMI） groups,and 50 healthy adults served as controls.The levels of the three biomarkers were detected by ELISA.The results showed that serum level of cystatin C（mg/L） was higher in CAD patients compared with those in the healthy controls（AMI vs.unstable angina pectoris vs.stable angina pectoris vs.controls：1.27±0.18 vs.1.09±0.19 vs.0.91±0.05 vs.0.78±0.07,all P〈0.01）.Cathepsin S（ng/mL） was also significantly different among the groups（AMI vs.unstable angina pectoris vs.stable angina pectoris vs.controls：67.30±8.36 vs.56.90±7.16 vs.49.8±2.72 vs.67.30±8.36,all P〈0.01）.IL-1（pg/mL） was significantly different among the groups as well（AMI vs.unstable angina pectoris vs.stable angina pectoris vs.controls：2.96±0.57 vs.2.46±0.24 vs.2.28±0.09 vs.2.02±0.13,all P〈0.01）.Spearman＇s correlation test revealed positive correlation between cystatin C,cathepsin S,IL-1 and Gensini score（r=0.451,0.491,0.397,respectively）.It is suggested that simultaneous detection of cystatin C,cathepsin S,and IL-1 in serum may be useful in clinical classification and assessment of severity of CAD.
