This study aimed to evaluate the milk clotting capacity (MCA), and caseinolytic activity (CA) of crude coagulant from the stomachs of adult rabbits (RC), and their influence on the composition, proteolysis, and textur...This study aimed to evaluate the milk clotting capacity (MCA), and caseinolytic activity (CA) of crude coagulant from the stomachs of adult rabbits (RC), and their influence on the composition, proteolysis, and texture of type Prato cheese. We tested two ways for salting the abomasum and three levels of pH and times for enzyme activation. The presence of enzymes and caseinolytic activity in RC was assayed by SDS-PAGE. The effects of three pH levels and the milk’s temperature on RC’s water-holding capacity (WHC) were evaluated. Brazilian Prato cheese type was developed and its proximate composition, texture, and change during ripening were assessed. The saturated NaCl solutions (37%, m/v) were more efficient for enzyme extraction from adult rabbit stomachs, and conditions of pH of 4.3 for 24 h showed better performance for enzyme activation into RC. An early and more active proteolytic reaction of RC on bovine caseinate was appreciated at the first 5 min of digestion when compared to commercial chymosin (CC). The water holding capacity of gel coagulated by RC was optimum at pH 6.4 and 37°C and is highly dependent on pH and temperature parameters. Total dry extract and fat were higher for treatment coagulated with RC. Conversely, hardness, gumminess, and chewability were lower in the same treatment. Yield, protein content, and changes that occurred during the ripening stage of cheeses were similar for both treatments, demonstrating that casein proteolysis was developed in a similar way. Coagulants from adult rabbits’ stomachs can be used to make semi-cooked curd cheeses.展开更多
BACKGROUND： Activated clotting factor VII has been demonstrated to exhibit obvious anti-apoptosis effects. OBJECTIVE： To observe the effect of activated clotting factor VII on neuronal apoptosis at different time po...BACKGROUND： Activated clotting factor VII has been demonstrated to exhibit obvious anti-apoptosis effects. OBJECTIVE： To observe the effect of activated clotting factor VII on neuronal apoptosis at different time points following rat intracerebral hemorrhage （ICH）. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Neurobiological Laboratory of Second Military Medical University from October 2005 to April 2006. MATERIALS： Recombinant-activated clotting factor Vlla （rFVtla） was purchased from Danish Novo Nordisk, Denmark. In situ cell death detection kit-POD kit was purchased from Roche, Switzerland. Caspase-3 activity determination kit from Biovision, USA. METHODS： A total of 72 healthy, male, Sprague Dawley rats, aged 5-8 months, were randomly assigned to three groups （n = 24）： sham-operated, ICH model, and rFVIla. In the ICH model and rFVIla groups, 80.0μL autologous non-clotting blood from rat tails was injected into the right caudate putamen to establish the ICH. The empty microinjector was inserted into the caudate putamen in the sham-operated group. The ICH model and rFVIla groups were subdivided into four subsets separately： 6, 24, 72 hours and 7 days following ICH. The rats in the rFVIla group were injected with 160 μg/kg rFVIla via the dorsal vein of the penis. MAIN OUTCOME MEASURES： Apoptotic cells were detected in the right caudate putamen by TUNEL; caspase-3 activity by spectrophotometry; and rat neurological function was evaluated by neurological functional impairment scales. RESULTS： Rat neurological function was deteriorated at 24, 72 hours, and 7 days following ICH. The TUNEL-positive cells and caspase-3 activity in the right caudate putamen was significantly increased in the ICH rats （P 〈 0.05）; rFVlla treatment reduced the number of TUNEL-positive cells and caspase-3 activity in the right caudate putamen （P 〈 0.05）, and neurological function was significantly improved （P 〈 0.05）. CONCLUSION： rFVIla was applied within 72 hours after tCH, which reduced the amount of neuronal apoptosis and promoted neurological function restoration by possibly inhibiting caspase-3 activity.展开更多
Background Due to lack of point-of-care testing, the use of low-molecular-weight heparin （LMWH） therapy in some special patients is restricted. This study was designed to explore the effects of LMWH on clot rate （C...Background Due to lack of point-of-care testing, the use of low-molecular-weight heparin （LMWH） therapy in some special patients is restricted. This study was designed to explore the effects of LMWH on clot rate （CR） and activated clotting time （ACT）, and to search for an appropriate method for bedside monitoring of anticoagulant activity of LMWH. Methods Thirty-two healthy volunteers were selected from the staff of Beijing Tongren Hospital. CR and ACT were measured with different reagents （glass beads, diatomite, kaolin and magnetic bar） on blood samples spiked with increasing concentrations of LMWH （dalteparin, 0.2-1.8 IU/ml）. Correlations between concentrations of LMWH and values of CR and ACT were analysed based on the data obtained and regression analysis was performed to establish a regression equation. Results With the increase in doses of dalteparin, CR values reduced gradually. The values of CR of four reagents （glass beads, diatomite, kaolin and magnetic bar） were 20.4-4.5 IU/min, 27.4-6.9 IU/min, 27.5-7.9 IU/min and 7.8-0.1 IU/min respectively and an linear relationship was observed between the CR values and dalteparin concentrations （P〈0.05）. The values of ACT were 173-615 seconds, 130-270 seconds, 123-226 seconds, 337-1411 seconds respectively, which showed a linear regression between the ACT values and dalteparin concentrations （P〈0.01）. Differences in slope of the regression curves of ACT were observed with all the reagents tested （glass beads 248.2 s/IU, diatomite 74.8 s/IU, kaolin 58.2 s/IU and magnetic bar 1112.2 s/IU, P〈0.01）. While the minimum concentration of dalteparin was 0.2 IU/ml, 0.4 IU/ml, 1.4 IU/ml and 0.2 IU/ml separately, the ACT values of the four coagulants （glass beads, diatomite, kaolin and magnetic bar） were beyond the normal limit and showed a noticeable increase respectively （P〈0.01）. Conclusions This study showed that there was an excellent linear relationship between the CR and ACT values and dalteparin concentrations for all the four reagents （glass beads, diatomite, kaolin and magnetic bar） in vitro. The sensitivity of different coagulation reagents to LMWH different. Choosing a suitable reagent, both CR and ACT were possible to be used as a convenient bedside test for LMWH.展开更多
Background Although low-molecular-weight heparin has replaced unfractionated heparin to become the primary anticoagulation drug for treatment of acute coronary syndrome, there is no convenient bedside monitoring metho...Background Although low-molecular-weight heparin has replaced unfractionated heparin to become the primary anticoagulation drug for treatment of acute coronary syndrome, there is no convenient bedside monitoring method. We explored the best laboratory monitoring method of low-molecular-weight heparins （enoxaparin, dalteparin, and nadroparin） by use of the Sonoclot coagulation analyzer to monitor the activated clotting time. Methods A total of 20 healthy volunteers were selected and 15 ml of fasting venous blood samples were collected and incubated. Four coagulants, kaolin, diatomite, glass bead, and magnetic stick, were used to determine the activated clotting time of the low-molecular-weight heparins at different in vitro anti-Xa factor concentrations. A correlation analysis was made to obtain the regression equation. The activated clotting time of the different low-molecular-weight heparins with the same anti-Xa factor concentration was monitored when the coagulant glass beads were applied. Results The activated clotting time measured using the glass beads, diatomite, kaolin, and magnetic stick showed a linear correlation with the concentration of nadroparin （r = 0.964, 0.966, 0.970, and 0.947, respectively）. The regression equation showed that the linear slopes of different coagulants were significantly different （glass beads 230.03 s/IU, diatomite 89.91 s/IU, kaolin 50.87 s/IU, magnetic stick could not be calculated）. When the concentration of the anti-Xa factor was the same for different low-molecular-weight heparins, the measured activated clotting time was different after the application of the glass bead coagulant. Conclusions The glass bead coagulant is most feasible for monitoring the in vitro anticoagulation activity of nadroparin The different effects of different low-molecular-weight heparins on the activated clotting time may be related to the different anti-11a activities.展开更多
文摘This study aimed to evaluate the milk clotting capacity (MCA), and caseinolytic activity (CA) of crude coagulant from the stomachs of adult rabbits (RC), and their influence on the composition, proteolysis, and texture of type Prato cheese. We tested two ways for salting the abomasum and three levels of pH and times for enzyme activation. The presence of enzymes and caseinolytic activity in RC was assayed by SDS-PAGE. The effects of three pH levels and the milk’s temperature on RC’s water-holding capacity (WHC) were evaluated. Brazilian Prato cheese type was developed and its proximate composition, texture, and change during ripening were assessed. The saturated NaCl solutions (37%, m/v) were more efficient for enzyme extraction from adult rabbit stomachs, and conditions of pH of 4.3 for 24 h showed better performance for enzyme activation into RC. An early and more active proteolytic reaction of RC on bovine caseinate was appreciated at the first 5 min of digestion when compared to commercial chymosin (CC). The water holding capacity of gel coagulated by RC was optimum at pH 6.4 and 37°C and is highly dependent on pH and temperature parameters. Total dry extract and fat were higher for treatment coagulated with RC. Conversely, hardness, gumminess, and chewability were lower in the same treatment. Yield, protein content, and changes that occurred during the ripening stage of cheeses were similar for both treatments, demonstrating that casein proteolysis was developed in a similar way. Coagulants from adult rabbits’ stomachs can be used to make semi-cooked curd cheeses.
文摘BACKGROUND： Activated clotting factor VII has been demonstrated to exhibit obvious anti-apoptosis effects. OBJECTIVE： To observe the effect of activated clotting factor VII on neuronal apoptosis at different time points following rat intracerebral hemorrhage （ICH）. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Neurobiological Laboratory of Second Military Medical University from October 2005 to April 2006. MATERIALS： Recombinant-activated clotting factor Vlla （rFVtla） was purchased from Danish Novo Nordisk, Denmark. In situ cell death detection kit-POD kit was purchased from Roche, Switzerland. Caspase-3 activity determination kit from Biovision, USA. METHODS： A total of 72 healthy, male, Sprague Dawley rats, aged 5-8 months, were randomly assigned to three groups （n = 24）： sham-operated, ICH model, and rFVIla. In the ICH model and rFVIla groups, 80.0μL autologous non-clotting blood from rat tails was injected into the right caudate putamen to establish the ICH. The empty microinjector was inserted into the caudate putamen in the sham-operated group. The ICH model and rFVIla groups were subdivided into four subsets separately： 6, 24, 72 hours and 7 days following ICH. The rats in the rFVIla group were injected with 160 μg/kg rFVIla via the dorsal vein of the penis. MAIN OUTCOME MEASURES： Apoptotic cells were detected in the right caudate putamen by TUNEL; caspase-3 activity by spectrophotometry; and rat neurological function was evaluated by neurological functional impairment scales. RESULTS： Rat neurological function was deteriorated at 24, 72 hours, and 7 days following ICH. The TUNEL-positive cells and caspase-3 activity in the right caudate putamen was significantly increased in the ICH rats （P 〈 0.05）; rFVlla treatment reduced the number of TUNEL-positive cells and caspase-3 activity in the right caudate putamen （P 〈 0.05）, and neurological function was significantly improved （P 〈 0.05）. CONCLUSION： rFVIla was applied within 72 hours after tCH, which reduced the amount of neuronal apoptosis and promoted neurological function restoration by possibly inhibiting caspase-3 activity.
文摘Background Due to lack of point-of-care testing, the use of low-molecular-weight heparin （LMWH） therapy in some special patients is restricted. This study was designed to explore the effects of LMWH on clot rate （CR） and activated clotting time （ACT）, and to search for an appropriate method for bedside monitoring of anticoagulant activity of LMWH. Methods Thirty-two healthy volunteers were selected from the staff of Beijing Tongren Hospital. CR and ACT were measured with different reagents （glass beads, diatomite, kaolin and magnetic bar） on blood samples spiked with increasing concentrations of LMWH （dalteparin, 0.2-1.8 IU/ml）. Correlations between concentrations of LMWH and values of CR and ACT were analysed based on the data obtained and regression analysis was performed to establish a regression equation. Results With the increase in doses of dalteparin, CR values reduced gradually. The values of CR of four reagents （glass beads, diatomite, kaolin and magnetic bar） were 20.4-4.5 IU/min, 27.4-6.9 IU/min, 27.5-7.9 IU/min and 7.8-0.1 IU/min respectively and an linear relationship was observed between the CR values and dalteparin concentrations （P〈0.05）. The values of ACT were 173-615 seconds, 130-270 seconds, 123-226 seconds, 337-1411 seconds respectively, which showed a linear regression between the ACT values and dalteparin concentrations （P〈0.01）. Differences in slope of the regression curves of ACT were observed with all the reagents tested （glass beads 248.2 s/IU, diatomite 74.8 s/IU, kaolin 58.2 s/IU and magnetic bar 1112.2 s/IU, P〈0.01）. While the minimum concentration of dalteparin was 0.2 IU/ml, 0.4 IU/ml, 1.4 IU/ml and 0.2 IU/ml separately, the ACT values of the four coagulants （glass beads, diatomite, kaolin and magnetic bar） were beyond the normal limit and showed a noticeable increase respectively （P〈0.01）. Conclusions This study showed that there was an excellent linear relationship between the CR and ACT values and dalteparin concentrations for all the four reagents （glass beads, diatomite, kaolin and magnetic bar） in vitro. The sensitivity of different coagulation reagents to LMWH different. Choosing a suitable reagent, both CR and ACT were possible to be used as a convenient bedside test for LMWH.
文摘Background Although low-molecular-weight heparin has replaced unfractionated heparin to become the primary anticoagulation drug for treatment of acute coronary syndrome, there is no convenient bedside monitoring method. We explored the best laboratory monitoring method of low-molecular-weight heparins （enoxaparin, dalteparin, and nadroparin） by use of the Sonoclot coagulation analyzer to monitor the activated clotting time. Methods A total of 20 healthy volunteers were selected and 15 ml of fasting venous blood samples were collected and incubated. Four coagulants, kaolin, diatomite, glass bead, and magnetic stick, were used to determine the activated clotting time of the low-molecular-weight heparins at different in vitro anti-Xa factor concentrations. A correlation analysis was made to obtain the regression equation. The activated clotting time of the different low-molecular-weight heparins with the same anti-Xa factor concentration was monitored when the coagulant glass beads were applied. Results The activated clotting time measured using the glass beads, diatomite, kaolin, and magnetic stick showed a linear correlation with the concentration of nadroparin （r = 0.964, 0.966, 0.970, and 0.947, respectively）. The regression equation showed that the linear slopes of different coagulants were significantly different （glass beads 230.03 s/IU, diatomite 89.91 s/IU, kaolin 50.87 s/IU, magnetic stick could not be calculated）. When the concentration of the anti-Xa factor was the same for different low-molecular-weight heparins, the measured activated clotting time was different after the application of the glass bead coagulant. Conclusions The glass bead coagulant is most feasible for monitoring the in vitro anticoagulation activity of nadroparin The different effects of different low-molecular-weight heparins on the activated clotting time may be related to the different anti-11a activities.
