3.Effects of the monitoring of cerebral blood flow in the preparation of cerebral ischemia model in rats
Long WANG ; Baozhe JIN ; Xinzhong ZHANG
Chinese Journal of Cerebrovascular Diseases 2017;14(5):254-260
Objective To investigate whether the monitoring of cerebral blood flow can be used to evaluate the successful preparation of rat models with focal cerebral ischemia.Methods With the line plug inserted into the left internal cerebral artery of 30 SPF Wistar Han rats at (16.0±0.5) mm,(18.0±0.5) mm and (20.0±0.5) mm respectively,three kinds of focal cerebral ischemia models (10 each) were prepared.All the rats were divided into incomplete occlusion group,complete occlusion group and deep occlusion group according to whether the blood clots were found in skull base and the infarction was appeared in the middle cerebral artery territory after the whole brain was stained with 2,3,5-triphenyl tetrazolium chloride.The cerebral blood flow in the middle cerebral artery territory of each animal was monitored by laser Doppler at the time of before and after blocking with the line plug inserted and reperfusion with the line plug pulled out and recorded for statistical analysis.The cerebral blood flow was expressed as the relative flow unit (perfusion unit,PU);The changes of the cerebral blood flow after occlusion and reperfusion were expressed as a percentage of that blood flow and those before occlusion.Results During the process of modeling,1 rats died,and the other in the incomplete occlusion group (n=9),in the complete occlusion group (n=15),and in the deep occlusion group (n=5).The depths of inserting of 8 rats in the incomplete occlusion group were about (16±0.5) mm,and the blood flow from the anterior cerebral artery to middle cerebral artery could not be prevented completely,the Longa score was evaluated 6 h after ischemia and ranged from 0 to 1,there were no blood clots in skull base and no infarction after TTC staining.The depths of inserting of 9 rats in the complete occlusion group were about (18±0.5) mm,and the anterior cerebral artery blood flow was completely blocked,the neurological dysfunction was significant after 6 h ischemia and the Longa score was ranged from 2 to 3,the blood clots in skull base were absent and the infarctions in middle cerebral artery territory were obvious after TTC staining.The depth of inserting of 5 rats in the deep occlusion group were about (20±0.5) mm,and the anterior cerebral artery blood flow was also completely blocked,the neurological dysfunction was serious after 6 h ischemia and the Longa score was ranged from 3 to 4,there existed blood clots in skull base and obvious infarctions in middle cerebral artery territory after TTC staining.The cerebral blood flows after the line plug inserted compared with those before occlusion in the incomplete occlusion group,complete occlusion group and deep occlusion group were all decreased (94±17 vs.256±36,43±9 vs.286±44,44±6 vs.294±46,respectively,all P<0.05) ,and the difference between groups was statistically significant (F=56.57,P<0.01),Those in complete occlusion group and deep occlusion group were lower than that in incomplete occlusion group(all P<0,05),but the difference between deep occlusion group with complete occlusion group was not statistically significant(P>0.05);The percentages of cerebral blood flow after occlusion and before were (36.93±0.06)%,(15.09±0.02)%,(15.52±0.04)% respectively,and the difference between groups was statistically significant (F=39.14,P<0.01).The cerebral blood flow after reperfusion were 213±31,147±17,96±14,respectively,and the difference was also statistically significant (F=50.05,P<0.01),and the cerebral blood flow in deep occlusion group was less than that in complete occlusion group(P<0.05);The percentages of cerebral blood flow after reperfusion and before occlusion were (83.10±0.02)%,(51.83±0.05)%,(33.49±0.09)% respectively,and the difference was also statistically significant (F=93.23,P<0.01).Conclusion The cerebral blood flow monitored by laser Doppler can be used as a real-time,convenient,micro invasive,objective and reliable standard to evaluate the successful preparation of rat MCAO models with line plug method.
4.The changes of serum IL-6,IL-10,PLA2 on intracranial infection and its relationship with prognosis
Long YANG ; Yulin ZHANG ; Fuqiang JIN
Chongqing Medicine 2015;(33):4655-4657
Objective To explore the changes of serum IL‐6 ,IL‐10 ,PLA2 on intracranial infection and its prognosis .Methods Totallly 100 patients with intracranial infection during February 2011 to December 2013 were selected as the research object .In 100 cases of intracranial infection patients ,30 cases were cured (group A) and 52 cases improved (group B) ,18 cases of illness or death as poor prognosis group (group C) .Serum IL‐6 ,IL‐10 ,PLA2 content of the subjects in different time point were detected .Results One day After infection ,serum IL‐6 ,IL‐10 ,PLA2 levels in intracranial infection group were obviously higher than that of healthy control group (P< 0 .01) ;7 d after infection ,IL‐6 ,IL‐10 average level among 3 groups had obvious changed(P< 0 .05) ,7 d after in‐fection ,IL‐6 ,IL‐10 level of group B and group C was significantly higher than group A (P< 0 .05) ;and in group C ,IL‐6 ,IL‐10 was higher than group B (P< 0 .05) .7 d after infection ,the IL‐6 IL‐ 10 level of group A declined ,the IL‐6 ,IL‐ 10 levels of group B be‐gan decreasing 14 d after infection ,and the IL‐6 ,IL‐10 levels of group C had been in a rising trend .3 d after infection ,the PLA2 leves among 3 groups had obvious changed(P< 0 .05) ,7 d after infection ,the change rate increased ,7 d after infection ,PLA2 level of group B and group C was significantly higher than group A (P< 0 .05) ,and in group C ,PLA2 was higher than group B (P< 0 . 05) .7 d after infection ,the PLA2 level of group A declined ,in group B and group C ,PLA2 level began to decline significantly 14 d after infection .Conclusion IL‐6 ,IL‐10 ,PLA2 are closely related to the occurrence and prognosis of intracranial infection .
5.Evaluation for the performance of Sysmex XN 20 A1 automation blood cell analyzer
Fang JIN ; Jiancheng LONG ; Chunxia ZHANG
China Medical Equipment 2016;13(8):31-34
Objective:To evaluate the performance for the Sysmex XN20 A1 automation blood cell analyzer.Methods: The precision, contaminative rate, linear range, blank, accuracy and various sample models were verified, and low values of blood platelet were compared with the methods of microscopy and dye.Results:The contaminative rate was lower than 0.4%. The linear arrange and precision were well for the analyzer. The values of RBC, HGB and PLT were within the range of 1±0.05 and the correlation coefficients were higher than 0.975. The bias of average and constant values by the accuracy verification samples test met the demands of accuracy. And the relative differences of various sample models meet the requirements of the comparability. The variable coefficient of low values of PLT was lower than 4% by the dye method.Conclusion: The Sysmex XN20 A1 automation blood cell analyzer has the characteristics of perfect precision, accuracy, low contaminative rate, broad linear arrange and good repeatability for the low level PLT. It can be applied in clinical laboratory.
7.Role of autophagy and apoptosis in tumor.
Jin-long TANG ; Hong-he ZHANG ; Mao-de LAI
Chinese Journal of Pathology 2012;41(8):573-576
Animals
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Antineoplastic Agents
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therapeutic use
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Apoptosis
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Apoptosis Regulatory Proteins
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metabolism
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Autophagy
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physiology
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Beclin-1
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Humans
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Membrane Proteins
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metabolism
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Neoplasms
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drug therapy
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metabolism
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pathology
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Signal Transduction
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TOR Serine-Threonine Kinases
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metabolism
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Tumor Suppressor Protein p53
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metabolism
10.Effect of hydrogen on activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion in rats
Feng JIN ; Long HE ; Yanqiu AI ; Da LI ; Wei ZHANG
Chinese Journal of Anesthesiology 2017;37(3):305-307
Objective To evaluate the effect of hydrogen on the activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups (n=12 each) using a random number table:sham operation (group S),I/R group and hydrogen group (group H).Cerebral ischemia was induced by occlusion of the middle cerebral artery followed by reperfusion in I/R and H groups.In group H,hydrogen-rich saline 5 ml/kg (0.6 mmol/L) was injected intraperitoneally at 3 days before establishment of the model and immediately after the onset of reperfusion.At 24 h of reperfusion,the rats were sacrificed,and hippocampal tissues were obtained for determination of neuroapoptosis (by TUNEL),apoptotic neuron count and expression of activated caspase-3 (by Western blot).The brain tissues in the ischemic area were obtained and stained with haematoxylin and eosin for examination of the pathological changes.Results Compared with group S,the expression of activated caspase-3 was significantly up-regulated,and the apoptotic neuron count was increased in I/R and H groups (P<0.05).Compared with group I/R,the expression of activated caspase-3 was significantly down-regulated,the apoptotic neuron count was decreased (P<0.05),and the pathological changes of brain tissues were significantly reduced in group H.Conclusion The mechanism by which hydrogen inhibits neuroapoptosis during cerebral I/R is probably related to inhibited activation of caspase-3 in brain tissues of rats.