1.Immunocytochemical study of phospholipase C-gamma1 expression in mouse embryonic tissue.
Jun LIU ; Ming LI ; Wei-lie HU ; Jun LÜ ; Hai-bo NIE ; Qing-rong LI ; Yu WANG ; Shen-qiu LUO
Journal of Southern Medical University 2006;26(8):1166-1169
To investigate the expression of phospholipase C-gamma1 (PLC-gamma1) in mouse embryonic tissues, serial tissue sections were prepared routinely for immunocytochemistry for PLC-gamma1. The results showed that PLC-gamma1 was expressed in the cartilage, skeletal muscles, myocardium, the collecting tubule of the kidney, connective tissues and the brain, suggesting the important role PLC-gamma1 and the related signal pathway may play in the development of mouse embryonic tissues.
Animals
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Brain
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embryology
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enzymology
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Cartilage
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embryology
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enzymology
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Embryo, Mammalian
;
enzymology
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Female
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Fetal Heart
;
enzymology
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Immunohistochemistry
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Kidney
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embryology
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enzymology
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Mice
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Muscle, Skeletal
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embryology
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enzymology
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Phospholipase C gamma
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biosynthesis
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Pregnancy
2.Studies of Hepatic, Brain Monoamine Oxidase and Brain Serotonin in Rats.
Yonsei Medical Journal 1984;25(1):27-38
The effects of 7-ethyl-8-methylf1avin (7-Et) and 7-methyl-8-ethyl-flavin (8-Et) on rat hepatic monoamine oxidase (MAO), brain MAO activity and 5-hydroxytryptamine (5-HT or serotonin) in rat brain were investigated. In the study of hepatic MAO activity, kynur-amine a nonphysiological substrate for both A and B type MAO, was used for a spectro-photometric method, and [14C]-labeled amines were also used for a radiometric procedure for camparison with MAO activity determined by the spectrophotometric method. The rate of change in MAO activity of hepatic mitochondria from rats receiving Rb-def and 7-Et and 8-Et flavin showed the activity was severely reduced during 8 weeks. Rapid reduction of enzyme activity (50% in def-group, 35% in 7-Et group and 8% 8-Et flavin group) was observed at the end of 2 weeks. The enzyme activity lasted with slow decre-ment of enzyme level from 4 weeks to the end of 8 weeks as low as 16% in def, 18% in 7-Et and 3% in 8-Et flavin group. The trend of decrement of MAO activity when kynura-mine was used as a substrate appears to be similar with the small variation of MAO activity when [14C]-labelled tyramine, dopamine, serotonin and tryptamine respectively were used as substrate. The rate of decay of brain mitochondrial MAO activity in rats receiving each respective f1avin was not rapid and severely depressed as the MAO activity we have found in liver mitochondrial MAO of rats during the 8 week experimental time, but a similar tendency of decay of MAO in each group was observed. The potent inhibitory effect of 8-Et on brain MAO was confirmed by the study of the simultaneous measure-ment of MAO activity in each experimental group. when the reduction Of brain MAO activity in rats receiving 8-Et after 6 weeks was approximately 80% of normal and in the same rats the concentration of brain 5-HT showed a 60% increment of that of the normal mts. During the experimental period there is no absolute parallelism between the MAO inhibition and 5-HT increase. However when the reduction of MAO activity reached 80% of normal value, the concentration of 5-HT increased dramatically as much as 60% of normal value. The results so far suggest clearly that 8-Et produces a much more potent inhibitory effect on the hepatic MAO a s well as brain MAO in rats. Therefore our present and previous results suggest that 7-Et and 8-Et flavin should bind itself to hepatic, brain MAO apoenzyme in the condition of total absence of riboflavin in these animals, and the holenzyme is catalytically inactive.
Animal
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Brain/enzymology*
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Brain/metabolism
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Comparative Study
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Male
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Mitochondria/enzymology*
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Mitochondria, Liver/enzymology*
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Monoamine Oxidase/metabolism*
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Rats
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Serotonin/metabolism*
3.Postburn change in the plasma level of neuron specific enolase in burned patients with cerebral malfunction.
Guang-qing WANG ; Zhen-guo WANG ; Shi-chu XIAO ; Zhao-fan XIA
Chinese Journal of Burns 2003;19(6):347-348
OBJECTIVETo explore the relationship between the change in neuron specific enolase (NSE) and brain malfunction in burned patients.
METHODSThe serum samples of 11 burned patients with brain dysfunction were collected for the development of the serum level of neuron specific enolase with radioimmunoassay, and the correlation between condition of systemic inflammation and the levels of neuron specific enolase was assessed.
RESULTSThe level of NSE in burn patients with cerebral malfunction was obviously higher than that in control, and the level was correlated with the systemic inflammation.
CONCLUSIONThe change in the level of serum NSE could reflect the damage degree of central nervous system to some extent.
Adult ; Brain Diseases ; enzymology ; Burns ; enzymology ; Humans ; Male ; Middle Aged ; Phosphopyruvate Hydratase ; blood ; Systemic Inflammatory Response Syndrome ; enzymology
4.The expression of phenylalanine hydroxylase in the brain of ragworm Neanthes japonica (Polychaeta, Annelida).
Guimin REN ; Zhe DONG ; Chao LIU ; Yimeng LIU ; Zhidong LUAN ; Qi LIU ; Xuexiang BAO ; Shun WANG
Chinese Journal of Biotechnology 2016;32(4):518-526
Phenylalanine hydroxylase (PAH) is a member of aromatic amino acid hydroxylase (AAAHs) family, and catalyze phenylalanine (Phe) into tyrosine (Tyr). Using immunological and RT-PCR methods to prove the existence of phenylalanine hydroxylase (PAH) gene in the brain of Neanthes japonica in protein and nucleic acid level. Using Western blotting to detect the pah immunogenicity of Neanthes japonica. Making paraffin sections and using immunohistochemical technique to identify the presence and distribution of the phenylalanine hydroxylase gene in the brain of Neanthes japonica. Clone pah gene from the brain of Neanthes japonica by RT-PCR, constructing plasmid and transferring into E. coli to amplification, picking a single homogeneous colony, double digesting then making sequence and comparing homology. Western blotting results showed that the expression of the protein is present in Neanthes japonica brain, immunohistochemistry technique results showed that phenylalanine hydroxylase mainly expressed in abdominal of forebrain, dorsal and sides of midbrain. RT-PCR technique results showed that the phenylalanine hydroxylase exist in the brain of Neanthes japonica and has a high homology with others animals. PAH is present in the lower organisms Neanthes japonica, in protein and nucleic acid level. Which provide the foundation for further study the evolution of aromatic amino acid hydroxylase genes in invertebrate.
Animals
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Blotting, Western
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Brain
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enzymology
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Escherichia coli
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metabolism
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Phenylalanine Hydroxylase
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genetics
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metabolism
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Polychaeta
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enzymology
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genetics
5.Transglutaminase and neurodegenerative diseases.
Zhen LIU ; Junsheng ZENG ; Sheng ZENG ; Beisha TANG ; Junling WANG
Chinese Journal of Medical Genetics 2015;32(4):562-566
Transglutaminase (TG) is a kind of calcium-dependent enzymes. The TGase family found in rodents and human contains 9 types, including TG1-7, blood coagulation factor XIIIa and erythrocyte membrane protein 4.2, with the former 8 types possessing catalytic activity. TG catalyzes various conversion reactions of glutamine, including transamination, deamination and esterification, and participates in post-transcriptional modification of proteins such as cross-linking peptides glutamine residue and lysyl-residue, stabilizing protein structure and catalyzing formation of protein aggregates. TGase has been found to contribute to a variety of important physiological and pathological processes and play a role in the pathogenesis of multiple diseases. Notably, neurodegenerative diseases such as Huntington's disease, spinocerebellar ataxia, Alzheimer's disease and Parkinson's disease, have a close connection with TGase's role in the human body.
Animals
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Brain
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enzymology
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Humans
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Neurodegenerative Diseases
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enzymology
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genetics
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Transglutaminases
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genetics
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metabolism
6.Regional Distribution of Lactate Dehydrogenase Isozymes in Rabbit Brain.
Yonsei Medical Journal 1980;21(1):1-8
The resolution of lactate dehydrogenase isozymes in tissue samples obtained from various regions of the rabbit brain was carried out by cellulose acetate electrophoresis. 1) Most regions of the brain showed an H-type isozyme pattern. 2) Five clear1y differentiated patterns of isozyme activity were found throughout the entire cerebral cortex with no difference between the lobes of the cerebral cortex. 3) All 5 patterns were found in the upper brain, while 4 patterns-LDH-1, 2, 3 and very 1ow activity of LDH-4 were found in the lower brain.
Animal
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Brain/enzymology*
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Isoenzymes
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Lactate Dehydrogenase/analysis*
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Rabbits
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Tissue Distribution
7.Expression, purification and crystallization of rat brain kinesin.
Qun WAN ; Pingting ZHU ; Houning LÜ ; Xinhong CHEN
Chinese Journal of Biotechnology 2014;30(3):485-491
Kinesin is a motor protein that uses the energy from ATP hydrolysis to move along the microtubule system. To investigate how the chemical energy stored in ATP is converted to mechanical movement, the corresponding N-terminal region of rat brain kinesin was expressed in BL21-Codon Plus (DE3)-RP competent cells. After SP-cation exchange chromatography and size exclusion chromatography, the protein yield reached 10 mg/L culture with the purity above 95%. The purified protein had ATPase activity and specifically reacted with the kinesin antibody in the Western blotting analysis. The purified kinesin was crystallized under the following condition: 1.7 mol/L (NH4)2SO4, 500 mmol/L NaCl, 20% glycerol. The kinesin crystal can diffract up to 2.0 angstroms resolution.
Animals
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Brain
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enzymology
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Crystallization
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Hydrolysis
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Kinesin
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chemistry
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isolation & purification
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Rats
8.nNOS expression of hippocampal neurons in aged rats after brain ischemia/reperfusion and its role in DND development.
Chuanhong YANG ; Huangwen LAI ; Chunlie ZHAN ; Yuhua XIAO ; Wenling ZHENG
Chinese Journal of Traumatology 2002;5(4):232-236
OBJECTIVETo study the role of neuronal nitric oxide synthase (nNOS) in aged rats' hippocampal delayed neuronal death (DND) following brain ischemia.
METHODSModels of incomplete brain ischemia were induced by clipping common carotid artery. A total of 46 aged SD rats were divided into 8 groups: normal control group (Group A, n=5), sham-operation group (Group B, n=5), reperfusion 1, 6, 12, 24, 48, and 96 hours groups after brain ischemia for 30 minutes (Group C, D, E, F, G, and H, n=6/group). The expression of nNOS was examined by immunohistochemistry and neuronal ultrastructural changes were observed by the transmission electron microscopy (TEM) at different time points after reperfusion.
RESULTSImmunohistochemistry showed that nNOS expression in the hippocampal neurons was high in Group E, low expression in Group D, moderate expression in Group F and G. There was nearly no expression of nNOS in Group A, B, C, and H. Ultrastructure of hippocampal neurons was damaged more severely in reperfusion over 24 hours groups.
CONCLUSIONSNitric oxide (NO) may be one of the important factors in inducing DND after ischemia/reperfusion.
Animals ; Apoptosis ; Brain Ischemia ; enzymology ; Female ; Hippocampus ; enzymology ; pathology ; Immunohistochemistry ; Male ; Microscopy, Electron ; Neurons ; enzymology ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; enzymology
9.Mechanisms of lysosomal proteases participating in cerebral ischemia-induced neuronal death.
Ai-Ping QIN ; Hui-Ling ZHANG ; Zheng-Hong QIN
Neuroscience Bulletin 2008;24(2):117-123
There are three different types of cell death, including apoptosis (Type I), autophagic cell death (Type II), and necrosis (Type III). Ischemic neuronal death influences stroke development and progression. Lysosomes are important organelles having an acidic milieu to maintain cellular metabolism by degrading unneeded extra- and intracellular substances. Lysosomal enzymes, including cathepsins and some lipid hydrolases, when secreted following rupture of the lysosomal membrane, can be very harmful to their environment, which results in pathological destruction of cellular structures. Since lysosomes contain catalytic enzymes for degrading proteins, carbohydrates and lipids, it seems natural that they should participate in cellular death and dismantling. In this review, we discuss the recent developments in ischemic neuronal death, and present the possible molecular mechanisms that the lysosomal enzymes participate in the three different types of cell death in ischemic brain damage. Moreover, the research related to the selective cathepsin inhibitors may provide a novel therapeutic target for treating stroke and promoting recovery.
Animals
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Apoptosis
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Autophagy
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Brain Infarction
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enzymology
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physiopathology
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Brain Ischemia
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enzymology
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physiopathology
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Cathepsins
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metabolism
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Humans
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Lysosomes
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metabolism
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Necrosis
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physiopathology
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Nerve Degeneration
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enzymology
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physiopathology
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Peptide Hydrolases
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metabolism
10.The subgroup change of nitric oxide synthase in earlier period with cerebral ischemia/reperfusion in rats.
Shang XIU-LI ; Liu JIA-HUI ; Xue YI-XUE
Chinese Journal of Applied Physiology 2010;26(2):144-146
OBJECTIVETo observe the expression of endothelial nitric oxide synthase (eNOS)and nervous nitric oxide synthase (nNOS) in rats during cerebral ischemia/reperfusion (CI/R) and study if change will be happen in subgroup between eNOS and nNOS during earlier period of CI/R.
METHODSA total of 60 Wistar rats weighting 200-280 g, supplied by Animal Center of China Medical University, were divided into 6 groups (n=10) (sham operation group; ischemia 1 h, 2 h group; reperfusion 0.5 h, 1 h, 2 h group). Female and male was half-and-half. Cerebral ischemia/reperfusion injury was induced by a 2-hour suture occlusion of the unilateral middle cerebral artery, immediately after suture withdrawal to allow reperfusion, eNOS and nNOS expressions were examined by the method of immunohistochemistry.
RESULTSeNOS expressions increased in 1-hour during ischemia, keeping up with decreasing until reperfusion 2-hour. While nNOS expressions increased in 2-hour between ischemia and reperfusion.
CONCLUSIONChanges of expression between eNOS and nNOS in rats during cerebral ischemia/reperfusion are different. This may be related with ischemia and reperfusion injury.
Animals ; Brain ; enzymology ; Brain Ischemia ; enzymology ; Female ; Male ; Nitric Oxide Synthase Type I ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury ; enzymology ; Time Factors