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Objective To observe BQ123 on lung tissue injury following ischemia-reperfusion of hind limbs in rats.Methods A total of 30 male SD rats were randomly divided into control,I-R,and BQ123 groups,with 10 rats in each group. After 4- hour of ischemia and 4-hour of reperfusion to the hind limbs, ET-1 was measured by radioimmunoassay, levels of myeloperoxidase (MPO) and malondialehyde (MDA) were examined by biochemical method,and the content of P-selectin was examined by ELISA. Immunohistochemical method was used to detect the expressions of Fas,Bcl-2 and Caspase-3 in lung tissues. Pulmonary apoptosis was examined by means of TdT-mediated dUTP nick end labeling ( TUNEL) . Results Compared with the control group,the levels of ET-1,MPO,MDA and P-selectin in lung tissues were all increased significantly in I-R group ( P<0. 01). The expressions of Fas,Bcl-2 and Caspase-3 were 0. 294±0. 003,0. 108±0. 005,and 0. 174±0. 003,significantly up-regulated in the I-R group. The apoptosis rate [(18. 83±2. 86)%] was significantly increased in the I-R group (P<0. 01). Compared with I-R group,the tissue contents of ET-1,MPO,MDA and P-selectin were significantly lowered in BQ123 group (P<0. 01). The expression levels of Fas (0. 115±0. 007) and Caspase-3 (0. 159±0. 006) were decreased,but the expression of Bcl-2 was increased (0. 128±0. 005). The apoptosis rate in BQ123 group was significantly lower [(10. 67±2. 16)%,P<0. 01].Conclusion BQ123 may have protective effects on lung tissue after limb ischemia-reperfusion in rats by means of improving neutrophil aggregation and reducing the expression of proteins related to cell apoptosis.
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We developed a three-dimensional mini-type permanent magnetic resonance imaging (MRI) device in our lab. The purposes of this study were (1) for further development of MRI technologies, (2) for support of broadening practices of animal test modeling in medical research, and (3) for training more specialists from colleges or universities in the field of MRI. This paper describes the research and development at our lab(s), especially stressing on the design of the main magnet, the gradient coil and the radio frequency coil. In addition, the specific methodologies used in our lab(s) and the related data are emphasized. The 3D MRI technologies have met the needs of using small animals, super thin sections of live animal body and high imaging resolutions. MRI images of mice head and abdominal have been obtained successfully by using the imager that we developed. The imaging results and analyses have also been discussed.
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Animals , Mice , Equipment Design , Imaging, Three-Dimensional , Magnetic Resonance Imaging , MethodsABSTRACT
Objective To observe the protection effect of ischemic preconditioning to small intestines injury after limbs ischemia reperfusion ,and investigate the role of nitrogen monoxidum in the process .Methods 52 male Wistar rats were randomly divided in-to Sham group ,limbs ischemia-reperfusion (IR) group ,IPC + IR group and Nω-nitro-L-arginine methylester (L-NAME) group . The value of diamine oxidase (DAO) and nitrogen monoxidum (NO) in plasma and gut tissue ,the content of fluorescein isothiocya-nate lipopolysaccharide (FITC-LPS) in plasma ,and the value of cyclic guanosine monophosphate (cGMP) and malondialehyde (MDA) in gut tissue were measured ,respectively .The expression of P-selection and Caspase-3 were measured by immunohisto-chemistry .The apoptosis of intestinal cells was detected by terminal deoxynucleotidyl transferase -mediated dUTP nick end labe-ling (TUNEL) and the results were analysed quantitatively by automatic image analytical system .Results Compared with the LIR group ,the values of DAO and FITC-LPS in plasma decreased obviously in the IPC group ,and the level of NO in plasma increased obviously (P<0 .01) .The values of DAO ,NO and cGMP in gut tissue increased obviously ,the level of MDA and the expression of Caspase-3 and the apoptosis index (AI) were significantly decreased in IPC group (P<0 .01) .Compared with the IPC group ,the value of DAO and FITC-LPS were higher and the level of NO was lower in plasma of the L-NAME group (P<0 .01) .The values of DAO ,NO and cGM P in gut tissue obviously decreased ,the level of MDA significantly increased ,and the expression of Caspase-3 and the AI significantly upregulated in L-NAME group than those in IPC group (P<0 .01) .Conclusion IPC can improve the small intestine injury and the apoptosis following hind limbs ischemia reperfusion which may correlated with the increase in the level of NO .
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AIM:To investigate the expression of angiotensin-converting enzyme 2 (ACE2) in nephritic epithelium of primates. METHODS:The expression of ACE2 in Vero E6 cells was detected by the techniques of RT-PCR and immunocytochemistry (ICC) techniques. The distribution of ACE2 protein in kidney tissues of two Rhesus monkeys and two normal human cases was observed by immunohistochemistry (IHC) techniques. RESULTS:Vero E6 cells were found to express both ACE2 mRNA and protein. ACE2 protein was mainly located in epithelium of proximal tubules of Rhesus monkey and human kidney. CONCLUSION:The expression of ACE2 in epithelium of primate kidney may provide the condition for SARS-CoV entry,which may be one of the reasons for inducing renal damage in SARS patients.
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Objective To observe the apeptosis in liver injury following limbs ischemia-reperfusion(IR) in rats and the protective effects of taurine.Methods The model of limbs ischemia-reperfusion injury was established.30 Wistar rats were randomized into 3 groups: control group,IR group and tanrine + reperfusion group (TR group) (n = 10 for each group).The levels of malondialdehyde (MDA) and xanthineoxidas (XOD), calcium and myeloper-oxidase (MPO) in the liver tissue were measured.DNA fragmentation was observed and analyzed by agarose gel e-lectrophoresis.Apoptosis was detected by TUNEL methods.The morphologic changes were observed with HE stai-ning.Results Compared with control group,the values of MDA, XOD, MPO, calcium in liver tissue were increased significantly in IR group (P<0.01), but the values of those in TR group were lower than in IR group (P<0.01).The percentage of apeptosis cell was higher in IR group than in control group(P<0.01).Compared with IR group, the percentage of apoptosis cell was lower in TR group (P<0.01).IR group presented DNA ladder pattern, while TR group showed no specific DNA ladder pattern in agarose gel electrophoresis.Conclusion Apoptosis participates in the liver injury after limb ischemia-reperfusion.Taurine can mitigate the liver injury and apoptosis after limb is-chemia-reperfusion injury in rats.
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BACKGROUND: Limb ischemia reperfusion (LIR) as a stressor leads to gastric mucosal injury, and then results in the occurrence of stress ulcer.OBJECTIVE: To observe the effects of LIR on gastric mucosal injury, investigate part of the mechanism, and the role of several transient limb ischemia in the occurrence of gastric mucosal injury.DESIGN: A randomized grouping design and controlled animal experiment.SETTING: Department of Pathophysiology of North China Coal Medical College.MATERIALS: The experiment was carried out in the pathophysiological laboratory of North China Coal Medical College from January to June 2002. Fifty-four healthy adult male Wistar rats were randomly divided into three groups with 18 rats in each group. Ischemia reperfusion (I/R) group: The rats were duplicated into models according to the Rosenthal method that under superficial anesthesia with ether, the roots of both hindlimbs were ligated by wrapping with rubber strap, blood flow was blocked for 4 hours and then recovered to perfusion for 4 hours, and finally killed by bleeding from abdominal aorta. Ischemic preconditioning group: Before model establishment, blood flow of both hindlimbs was blocked for 5 minutes, and then recovered to perfusion for 5 minutes, which was repeated for four times, and the following operations were the same as those in the I/R group. Control group: The operations were the same as those in the I/R group,but both hindlimbs were ligated at relaxation without blocking the blood flow.METHODS: Sections of gastric mucosa were prepared, and then observed under light microscope and electron microscope, and the index of gastric mucosal injury was determined according to the Guth standard. The colorimetric assay was performed with 721 spectrophotometer at 650 nm, and the amount of gastric combining mucus was calculated.Meanwhile, the blood flow of gastric mucosa, contents of phospholipid and hexosamine in gastric mucus, content of nitric oxide in plasma and gastric tissue and activity of nitric oxide synthase (NOS) in gastric mucosa were determined.MAIN OUTCOME MEASURES: Index of gastric mucosal injury, amount of gastric combining mucus, blood flow of gastric mucosa, contents of phospholipid and hexosamine in gastric mucus, contents of nitric oxide in plasma and gastric tissue and NOS activity in gastric mucosa.RESULTS: All the 54 rats were involved in the analysis of results. ① In the I/R group, gastric mucosal injury was serious, edema, hyperemia, erosion and disintegration of gland of mucosal glands were observed, infiltration of inflammatory cells (formation of ulcer) was observed in basal and inferior mucosa. In the ischemic preconditioning group, the gastric mucosa was complete, and the damaged severity was milder than that in the I/R group; Under electron microscope, the organell structures of gastric parietal and chief cells were incomplete and destroyed. The cell injuries in the ischemic preconditioning group were milder than those in the I/R group (index of injury: 18.00±10.71, 34.00±15.01, P＜ 0.01). ② The blood flow and combining mucosal amount of gastric mucosa, contents of phospholipid and hexosamine in gastric mucus in the ischemic preconditioning group and I/R group were all obviously lower than those in the control group [(2.12±0.56), (10.84±2.56), (25.52±2.97) mL/(kg·h); (2.01±0.91), (2.79±0.73), (3.99±0.87) mg;(7.68±1.95), (9.74±1.04), (11.98±1.98) mg/g; (3.83±1.18), (5.42±0.47), (5.76±1.21) mg/g, P ＜ 0.05, 0.01], those the above indexes were all higher in the ischemic preconditioning group than in the I/R group. ③ The contents of nitric oxide in plasma and gastric tissue and NOS activity in gastric mucosa in the ischemic preconditioning group and I/R group were significantly lower than those in the control group [(250.0±5.6), (270.0±11.3), (210.0±7.4) μmol/L; (9.34±0.67), (11.34±1.00), (7.50±0.67) μ kat/g, P ＜ 0.01], those were also signficantly higher in the ischemic preconditioning group than in the I/R group.CONCLUSION: As a stressor, LIR can lead to gastric mucosal injury, and cause stress ulcer.Ischemic preconditioning can alleviate the gastric mucosal injury following LIR
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Objective To investigate the effects of distractions on number processing using event-related potentials (ERPs). Methods ERPs were recorded in fourteen undergraduate students while they were asked to compare a heard number with the number "5" and judge whether its numerical value was larger or smaller than five. This task was carried out under a distracted attention (DA) condition and a focused attention (FA) condition. The reaction time and EEG signals were recorded during the experiment. Results There were significant differences between the reaction times in the different conditions (t26 = 3. 70,P
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AIM To evaluate the possible role of endogenous nitric oxide (NO) in the development of lung injury after hind limbs ischemia reperfusion (LIR), and the effect of taurine on endogenous NO on the model of lung injury following LIR. METHODS Wistar rats were divided into three groups: control group, ischemia reperfusion group (IR) and taurine+IR (Tau+IR). The changes of PaO 2 ,PaCO 2,lung index (LI) and lung permeability index (LPI) were observed. The content of malondialdehyde (MDA), NO and endothelin (ET) in both plasma and the lung as well as the content of nitric oxide synthase (NOS) and taurine in the lung were detected. The immunohistochemical method was used to detect level in the expression of NOS protein. RESULTS Taurine improved the pulmonary respiratory following LIR significantly, lightened lung edema, decreased the content of ET in both plasma and lung tissue, increased the content of NO in both plasma and lung tissue and promoted the level in the expression of NOS protein. CONCLUSION Using taurine may lighten lung injury following LIR and this protective effect may due to increasing the content of endogeneous NO and decreasing the level of ET.
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Objective To investigate cell injury and apoptosis of L-6TG cells induced by simulated ischemia-reperfusion,and to explore its regulating mechanism.Methods Cultured L-6TG cells were randomly divided into control group(C)and simulated ischemia-reperfusion group(IR).The L-6TG cells of C group were cultivated under normal condition in mimicking reperfusion solution,while the L-6TG cells of IR group were cultivated in mimicking ischemic solution for 4 hours,and then cultivated in mimicking reperfusion solution for 4 hours.Superoxide dismutase(SOD),xanthine oxidase(XOD)and free calcium in L-6TG cells were determined in each group;mitochondrial respiration was measured by MTT method.Apoptosis was identified by flow cytometer with propidium iodide(PI)staining method and agarose gel electrophoresis;the immunohistochemical method was used to determine the expression of bcl-2 and caspase-3,and their profiles were half-quantitatively analyzed by automatism image analytical system.Results Compared with the L-6TG cells of C group,after suffering from ischemia and reperfusion each for 4 hours,XOD,free calcium in L-6TG cells and apoptotic percentage were all increased significantly(287.88?8.47 vs 42?3.94 U/g prot,1.547?0.017 vs 7.23?0.28 lnMCF,7.23%?0.28% vs 0.69%?0.07%,P
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AIM: To study the distribution of nitric oxide synthase (NOS) in normal lung of rats, and the changes in distribution and activity of NOS in lung of rats suffering from limb ischemia reperfusion (LIR). METHODS: Using tourniquet, the model rats undergoing ischemia reperfusion were made. Using ?-ANDPH-d histochemical method, computer analysis of image and spectrophotography, the distribution of NOS in normal lungs of rats, and the changes in distributon and activity of NOS in lung of rats suffering from limb ischemia reperfusion (LIR) were observed. RESULTS: Histochemically, in normal rats, the NOS positive staining was found in epithelial cells of respiratory duct including bronchi, alveolar duct as well as in endothelium of pulmonary vessels, and the negative staining was found in pneumonocytes. In LIR rats, the above NOS staining were significantly stronger than that in normal one, and positive staining was also found in vascular smooth muscle and pneumonocytes. Biochemically, the results showed that the values of lung NOS, NO in LIR group increased significantly compared with the normal control. CONCLUSION: NO may play an important role not only in physiology of lung but also in pathophysiology of acute lung injury following LIR. [
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AIM: To observe the degree of gastric mucosal injury following limb ischemia/reperfusion(LIR),and to investigate the mechanism of gastric mucosal injury and the influence of the series of brief ischemia/reperfusion of hind limbs of rats on gastric mucosal injury. METHODS: Referring to Rosenthal's method, the model rats which underwent 4 hours of ischemia and 4 hours of reperfusion in hind limbs were made. The indexes of gastric mucosal injury after LIR and ischemic preconditioning (IPC) + LIR were determined. The morphologic changes were observed with light microscope and transmission electron microscop respectively. The GMBF,histologic lesion score, gastric barrier mucus, phospholipids, hexosamine, nitric oxide and nitric oxide synthase in mucus were measured in different groups. RESULTS: Serious damage in gastric mucosa was observed under microscope and EM after LIR. But less serious damage was observed in the IPC group. After LIR, compared to the control group, the GMBF and the content of gastric barrier mucus, phospholipids and hexosamine in mucus decreased significantly. There was significant difference in most indexes between the control group and the IPC group, but compared to LIR group, the histologic lesion score decreased significantly and the GMBF and the content of gastric barrier mucus, phospholipids, hexosamine ,nitric oxide and nitric oxide synthase in mucus increased significantly. CONCLUSION: LIR caused the gastric mucosa injury. IPC alleviated the damage of gastric mucosa following ischemia/reperfusion in hind limbs.
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In the study we determined the content of malondialdehyde(MDA) , which is the metabolite of lipid peroxide(LPO) , in plasm and lung homogenate by thiobarbituric acid(TBA) colorimetry, and the content of phosphatidyl-choline(PC), which is the major functional composition of pulmonary surfactant (PS).and total phospholipid (TPL) by thin-layer chromatogra-phy and inorganic phosphorus quantitative analysis in rabbits with lung injury induced by oleic acid. The results showed that the MDA level wasincreased in plasm and lung homogenate; PC and TPL were decreased in lung homogenate. Administrating of 654-2 before and after (0. 5 h)lung injury could both inhibit the production LPO and prevent the decrease of PS. The findings suggest that the decrease of PS may be due to the increase of LPO and that G54-2 could modify the process.
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Aim To investigate the effect of L-arginine(L-Arg) on myocardium in rat which suffered from limb ischemia-reperfusion.Methods Models of limb ischemia reperfusion were made by tourniquet methods.L-Arg(150 mg?kg-1) were intravascularly injected before reperfusion.Contents of malondialdehyde(MDA),myeloperoxidase(MPO) and tumour necrosis factor(TNF-?) in myocardium were measured.Levels of creatine kinase(CK),creatine kinase MB isoenzyme(CK-MB) and nitric oxide(NO) in plasma were deter-mined.The mean arterial pressure(MAP,left venteicular systolic pressure(LVSP),maximal rise rate of left venteicular pressure(dp/dtmax)and the maximal fall rate of left venteicular pressure(-dp/dtmax)were monitored.Morphologic changes of myocardium were evaluated after reperfusion.Results After rats' limbs suffering from ischemia-reperfusion,levels of MDA,MPO and TNF-? in myocardium,CK,CK-MB and NO in plasma increased differently(P
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AIM: To investingate the potential role of apoptosis in the development of acute lung injury(ALI) following limbs ischemia-reperfusion(LIR) in rats and the effects of raurine. METHODS: The model of LIB injury in rats was made. By using TUNEL, electrophoresis, reverse-transcription polymerase chain reaction(RT-PCR) and immunohistochemistry techniques, pulmonary apoptosis, Fas/FasL expressions and Ca~(2+), reactive oxygen species (ROS),the ratio of DNA double chain in tissue were detected in different groups.RESULTS: The rapid appearance of apoptosis in alveolar epithelial cells and pulmonary vascular endothelial cells were observed after LIR in rats. The ratio of DNA double chain decreased and tissue Ca~(2+) increased. The expression of Fas/FasL mRNA and protein was up-regulated in lung tissue of rats after LIR, which was related to the elevation of alveolar epithelial cells and pulmonary vascular endothelial cell apoptosis. Taurine decreased alveolar epithelial cell and pulmonary vascular endothelial cell apoptosis not by down-regulating expression of Fas/FasL system. CONCLUSIONS: These results suggest that the excessive apoptosis and Fas/FasL system expression may play a role in the pathogensis of ALI following LIB in rats, and taurine decreases alveolar epithelial cell and pulmonary vascular endothelial cell apoptosis.