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Objective To study the effect of Hydrogen on ischemia/reperfusion(I/R)-induced cardiocyte apoptosis and apoptosis related proteins expression in diabetic rats .Methods Diabetes mellitus was induced by intraperitoneal injection of streptozotocin (STZ) ,then feed 4 weeks before build ischemia/reperfusion model .60 SD male rats ,weight during 300-350 g ,were randomly divid-ed to six groups :non-diabetic sham-operated group (NS) ,non-diabetic I/R group(NI/R) ,non-diabetic hydrogen treated group (NH) ,diabetic sham-operated group(DS) ,diabetic I/R group(DI/R)and diabetic hydrogen treated group(DH) ,each group has 10 rats .The cardiac muscle I/R model was made by 30 min occlusion of the left anterior descending coronary artery and 2 h reperfu-sion .The rats in Hydrogen group were treated with 5 mL/kg hydrogen by intraperitoneal administration at the beginning of reper-fusion .The apoptosis index (AI) was calculated by TUNEL .The positive expressions of Bcl-2 ,Bax ,Caspase-3 in cardiomyocytes were respectively detected by immunohistochemistry .Results The apoptotic rates of cardiomyocytes and the positive expressions of Bcl-2 ,Bax ,Caspase-3 were significantly increased(P<0 .01) ,Compared with I/R group ,the apoptotic rates of cardiomyocytes in hydrogen treatment group were obviously decreased(P<0 .01) ,and the positive expressions of Bcl-2 were increased(P<0 .01) ,at the same time ,the positive expressions of Bax ,Caspase-3 were decreased(P<0 .01) .Conclusion Hydrogen inject by intraperitoneal method on myocardial ischemia-reperfusion injury of diabetic rats has a protective effect ;Its mechanism may be related to its inhibi-ting myocardial apoptosis by advanced the Bcl-2 protein expression and reduced the Bax 、Caspase-3 protein expression .
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Objective To evaluate the role of phosphoinositide 3 kinase/protein kinase B (PI3K/Akt) signaling pathway in hydrogen-induced inhibition of cell apoptosis during myocardial ischemia/reperfusion (I/R) in rats.Methods Forty healthy male Sprague-Dawley rats,weighing 300-350 g,were randomly allocated into 4 groups (n =10 each) using a random number table:sham operation group (S group),I/R group,hydrogen group (group H),and hydrogen + LY294002 group (group HL).Myocardial I/R was induced by occlusion of the anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion.In H and HL groups,99.6 ％ hydrogen 5 ml/kg was injected intraperitoneally immediately after beginning of reperfusion,and in addition LY294002 (0.3 mg/kg) was injected through the caudal vein before hydrogen injection in group HL.Arterial blood samples were collected at the end of 120 min reperfusion for determination of serum creatine kinase isoenzyme-MB (CK-MB) and lactate dehydrogenase (LDH) activities.The rats were then sacrificed.Myocardial apoptosis was detected by TUNEL and apoptosis index (AI) was calculated.The expression of Bcl-2,Bax and caspase-3 was detected by immuno-histochemistry.Bcl-2/Bax ratio was calculated.Results The serum CK-MB and LDH activities and AI were significantly increased,the expression of myocardial Bcl-2,Bax and caspase-3 was upregulated,and the ratio of Bcl-2/Bax was decreased in group I/R as compared with group S.Compared with group I/R,the serum CK-MB and LDH activities and AI were significantly decreased,the expression of myocardial Bcl2 was up-regulated,while the expression of myocardial Bax and caspase-3 was down-regulated,and the ratio of Bcl-2/Bax was increased in group H,and no significant changes were found in the parameters mentioned above in group HL.The serum CK-MB and LDH activities and AI were significantly increased,the expression of myocardial Bcl-2 was down-regulated,while the expression of myocardial Bax and caspase-3 was up-regulated,and Bcl-2/Bax ratio was decreased in group HL as compared with group H.Conclusion Hydrogen can activate the PI3K/Akt signaling pathway,and further up-regulates Bcl-2 expression and down-regulates Bax and Caspase-3 expression,thus inhibiting cell apoptosis during myocardial I/R in rats.
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ObjectiveTo investigate the effects of postconditioning with Shenmai-injectio on myocardial ischemia-reperfusion (I/R) injury in rats.MethodsThirty-six healthy male SD rats aged 10-12 weeks weighing 240-260 g were randomly divided into 3 groups ( n =12 each):sham operation group (group S) ; myocardial I/R group and Shenmai-injectio postconditioning group (group SPO).Myocardial I/R was produced by ligation of the left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion in groups I/R and SPO.In group SPO Shenmai-injectio 9 ml/kg was injected iv at the end of 30 min ischemia.Blood samples were collected from abdominal aorta at the end of 120 min reperfusion for determination of serum CK activity and cTnI concentration.The animals were then sacrificed.Myocardial specimens were obtained for microscopic examination,detection of apoptosis and determination of myocardial Bcl-2 and Bax protein expression ( by immuno-histochemis-try).ResultsMyocardial I/R significantly increased serum CK activity,cTnI concentration,apoptotic index (percentage of apoptotic cells) and Bax protein expression and decreased Bcl-2 protein expression in group I/R as compared with group S.Shenmai-injectio postconditioning significantly attenuated I/R-induced above changes and ameliorated histo-pathological damage in group SPO as compared with group I/R.ConclusionShenmai-injectiopostconditioning can reduce myocardial I/R injury by up-regulating Bcl-2 expression and down-regulating Bax expression,leading eventually to reduction in apoptosis.
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Objective To investigate whether pregnant rats exposure to ketamine cause offspring changes in space cognitive abilities and exploration abilities.Methods 3-month Sprague-Dawley female rats ( n =24)were randomly divided into four groups:group N (control group),group K1 (small doses of ketamine group),group K2 ( clinical anesthesia dose of ketamine group),group K3 ( large doses of ketamine group).3-month Sprague-Dawley male rats ( n =4) and female rats were mated at the same cage by the proportion of 2∶ 1.Pregnant mice were treated at tenth day:group N were treated saline with equal-volume to ketamine vein injection; group K1,group K2,group K3 administered vein injection 3,8,20mg/kg of ketamine.Then the 20-day offspring rats'learning and memory were assessed used Open Field Test ( record the time of the offspring in the central case through the number of grid within 2 min ) and Hole Board Test ( Counting the times of offspring stretch into the hole in 5 min) at postnatal days 20.Results In the Open Field Test,the retention time in central check of group N,group K2 and group K3 were (2.45 ± 1.23)s,(6.42 ±2.50)s,(6.41 ±2.19)s.Compared with group N,the retention time in central check of group K2 and group K3 were significantly higher (F=13.42,P＜0.01 ),and group K1 were not significant different ( t =1.33,P＞0.01 ),and the locomotion of group K1,group K2,group K3 were significantly reduced( ( 15.33 ± 6.81 ),( 13.75 ± 5.93 ),( 16.92 ± 6.54 ),F =4.24,P ＜ 0.05 ).In the Hole Board Test,the times of offspring stretch into the hole were not significant different comparing with the control group(F=2.17,P ＞ 0.05 ).Conclusion The dose of ketamine that equivalented clinical anesthesia can affect offspring rats' space cognitive abilities; but the exploring cognitive ability were not significantly influenced.
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ObjectiveTo study the safety and effectiveness of acute hypervolemic hemodilution (AHH) combined with tranexamic acid(TA) in neurosurgical operation. Methods Forty patients underwent selective neurosurgical operation were divided into two groups by radom digits table with 20 cases each, both groups were infused HES(130/0.4) 20 ml/kg for AHH after anesthesia, TA intravenous injection of loading 10 mg/kg, 1 mg/(kg·h) continuous infusion until the end of surgery in experimental group,only for AHH in control group. The mean arterial blood pressure(MAP), central venous pressure(CVP), heart rate (HR), pulse oxygen saturation (SpO2) were measured before AHH (T0), A HH immediately (T1), 1 h after AHH (T2), at the end of operation (T3), and in the corresponding time hemoglobin (Hb), hematocrit (Hct),prothrombin time (PT), activated partial thromboplastin time (APTT), platelet ( Plt ), fibrinogen ( FIB ) were measured; the intraoperative bleeding, blood transfusion and transfusion rate were counted in both groups.ResultsThere was no significant difference in HR and MAP at different time between two groups (P ＞0.05),compaued with T0,CVP at T1,T2 was increased (P＜0.05),Hb and Hct at T1,T2,T3 were decreased (P＜ 0.05 ). PT and APTT at T2,T3 were longer than that at T0 in control group (P＜ 0.05 ) ;Plt at T1 ,T2,T3 was lower in control group than that at T0 in two groups (P ＜ 0.05 ), Plt at T2, T3 was obviously increased in control group compared with experimental group (P ＜ 0.01 ) ; FIB at T2, T3 was lower than that at T0 in control group (P ＜0.05). The intraoperative bleeding, blood transfusion and transfusion rate in experimental group [(650 ±560) nl, (150 ± 50)ml,30％(6/20)]were lower than those in control group [(820 ±410) ml,(380 ±290) ml,60％ (12/20)],there were significant differences between two groups(P ＜0.05).ConclusionAHH combined with TA has obvious effect of saving blood and hemodynamic stability with less influence on coagulation in neurosurgical operation.
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Objective To observe the protection of aprotinin, ulinastatin and aminomethylbenzoic acid, aminomethylbenzoie acid on blood fibrinolytie system during cardiopulmonary bypass(CPB). Methods Thirty-six patients with rheumatic heart disease who were treated by valve replacement were randomly divided into three groups: aprotinin group (group A, 12 cases): aprotinin 2000 kU was added into the priming solution; ulinastatin and aminomethylbenzoic acid group (group UP, 12 cases): ulinastatin 12 000 U/kg and aminomethylbenzoic acid 10 mg/kg was added into the priming solution; aminomethylbenzoie acid group (group P, 12 cases): aminomethylbenzoic acid 10 mg/kg was added into the priming solution. Results There was no significant difference in CPB time and blood transfusion among three groups; the postoperative 24 h chest tube drainage in group UP was (443.3 ± 150.8) ml, in group P was (430.0 ± 178.3) ml and in group A was (290.0 ± 98.0) ml, there were significant differences between group UP, group P and group A (P < 0.05). There was 1 case of severe allergic reaction in group A. Conclusion Aprotinin, ulinastatin and aminomethylbenzoic acid, aminomethylbenzoic acid are effective in stabilizing blood fibrinolytic system and preserving platelet function during CPB, leading to less postoperative blood loss.
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Objective To investigate the effects of brain-derived neurotrophic factor (BDNF) pretreatment on neuron apoptosis and the expression of Bcl-2 and Bax protein following global cerebral ischemia-reperfusion (I/R) injury in gerbils. Method Forty-eight mongolian gerbils were randomly divided into six groups in equal number (n = 8): normal control group (group C), ischemia-reperfusion group (group I/R) and four BDNF pretreatment groups according to various lengths of time from BDNF pretreatment to ischemia-reperfusion. The BDNF pretreat-ment was carried out in gerbils with lateral ventricular injection of BDNF 0.5μg 6 h, 12 h,24 h and 48 hours be-fore cerebral ischemia, and those gerbils assigned into PR6, PR12, PR24 and PR48 groups. The global cerebral is-chemia-reperfusion was induced by occlusion of bilateral common carotid arteries for 20 minutes and then the arter-ies were released for 24 hours reperfusion. The confirmation of global cerebra ischemia was evidenced by the ap-pearance of mydriasis and disappearance of light reflex and righting reflex. Twenty-four hours later, all gerbils including those of control group were sacrificed and a piece of tissue was taken from frontal cortex just behind the optic chiasma 1～4 millimeter for making paraffin sections. Neuron apoptosis was identified by using TUNEL and immunohischemistry was used to detect the expression of Bcl-2 and Bax protein in cerebral cortex. The data were analyzed by using analysis of variance. Results There were no apoptotic cells, and expression of Bcl-2 and Bax protein positive cells found in group C. Neuron apoptosis in brain cortex was detected in I/R group and BDNF pre-treatment groups. The indexes of neuron apoptosis in BDNF pretreatment groups were markedly lower than those in group I/R (P < 0.01). Compared with group I/R, the index of expression of Bcl-2 protein positive cells was in-creased significantly in BDNF pretreatment groups (P = 0.005), while the index of expression of Bax protein posi-tive cells were decreased significantly (P < 0.01 in all groups). Among 4 BDNF pretreatment group, the lowest apoptosis index and lowest of expression of Bax protein positive cells were found in PR6 and PR12 BDNF pretreat-ment groups (P = 0.0056 and 0.001, respectively). Conclusions Different time windows of BDNF pretreatment can decrease the neuron apoptosis in different degree, and protect brain against cerebral ischemia-reperfusion injury significantly. Among BDNF pretreatment time windows, pretreatment of 6 hours and 12 hours are the better ones.The mechanism of protection of BDNF pretreatment may be attributed to inducing Bcl-2 protein expressions and in-hibiting Bax protein expressions, and thereby inhibiting neuron apoptosis.
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OBJECTIVE:To prepare internal eye perfuse aqua and establish a method for its quantity control.METHODS: The internal eye perfuse aqua was prepared in a germ free preparation standards,assaying was performed,and indexes like stability,etc.were investigated.RESULTS:The total chlorine amount,labeled amounts of magnesium and calcium in perfuse aqua all stood at 98.9%～100.6%.CONCLUSIONS:The preparation is reasonable in performulation,and simple,accurate and feasible in quality control,the preparation is stable and deserves to be popularized in the clinic.