1.Role of C5b-9 expression in skeletal muscle blood vessels in necrotizing myopathy.
Lu CONG ; Chuanqiang PU ; Yanling MAO ; Jiexiao LIU ; Xianghui LU ; Qian WANG
Journal of Southern Medical University 2012;32(5):714-717
OBJECTIVETo investigate the expression of C5b-9 in the skeletal muscle blood vessels in patients with necrotizing myopathy and explore its role in the pathogenesis of this disease.
METHODSThe expression of C5b-9 and MHC-I in the skeletal muscular fibers and blood vessels in 4 patients with necrotizing myopathy was detected using enzymohistochemistry and immunohistochemistry.
RESULTSFocal or dispersive necrotic muscle fibers with obvious phagocytosis were observed in all the 4 patients. No inflammatory cell infiltration was found in the perimysium or perivascular regions. HE staining showed a decreased number of local small blood vessels, and the some small blood vessels showed thickened vascular walls. Immunohistochemistry detected prominent C5b-9 expression in the necrotic muscle fibers and the blood vessels, and diffuse strong C5b-9 expression was found in the vascular walls, vascular endothelial cells and the smooth muscle layer. No MHC-I deposition was detected in the muscular fibers and blood vessels.
CONCLUSIONC5b-9 contributes to the pathogenesis of necrotizing myopathy mediated by pathologies in the blood vessels.
Aged ; Complement Membrane Attack Complex ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; blood supply ; Muscular Diseases ; blood ; pathology ; Necrosis
2.Diagnostic Significance of BAT in Anaphylaxis to Non-ionic Contrast Media.
Hao-yue ZHANG ; Su-jun XU ; Xiao-xian TANG ; Ji-jun NIU ; Xiang-jie GUO ; Cai-rong GAO
Journal of Forensic Medicine 2015;31(3):188-190
OBJECTIVE:
To investigate the diagnostic significance of basophil activation test (BAT) in anaphylaxis to non-ionic contrast media through testing the content of CD63, mast cell-carboxypeptidase A3 (MC-CPA3), and terminal complement complex SC5b-9 of the individuals by testing their levels in the normal immune group and the anaphylaxis groups to β-lactam drugs and non -ionic contrast media.
METHODS:
The CD63 expression of basophilic granulocyte in blood was detected by flow cytometry. The levels of MC-CPA3 in blood serum and SC5b-9 in blood plasma were detected by ELISA.
RESULTS:
The CD63 expression of basophilic granulocyte in blood, the levels of MC-CPA3 and SC5b-9 of anaphylaxis to non-ionic contrast media and β-lactam drugs were significantly higher than that in normal immune group (P < 0.05).
CONCLUSION
There is activation of basophilic granulocytes, mast cells and complement system in anaphylaxis to non-ionic contrast media. BAT can be used to diagnose the anaphylaxis to non-ionic contrast media.
Anaphylaxis/diagnosis*
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Basophils/cytology*
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Carboxypeptidases A/metabolism*
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Complement Membrane Attack Complex/metabolism*
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Contrast Media
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Flow Cytometry
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Granulocytes/cytology*
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Humans
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Mast Cells/cytology*
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Tetraspanin 30/metabolism*
3.The expression of Smac and XIAP in rat hippocampus following limbic seizure induced by kainic acid injection into amygdaloid nucleus.
Tian-Fu LI ; Yu-Min LUO ; Chuan-Zhen LU
Acta Physiologica Sinica 2004;56(2):172-177
To determine whether Smac/DIABLO (second mitochondrial activator of caspases/direct inhibitor of apoptosis protein-binding protein of low isoelectric point [PI]) and XIAP (X-chromosome-linked inhibitor of apoptosis protein) serve to regulate neuronal apoptosis following seizures, we investigated seizure-induced changes in caspase-9, Smac/DIABLO and XIAP protein expression and the in vivo effect of caspase-9 inhibition. Animals received unilateral intra-amygdaloid injection of kainic acid (0.5 microg) to induce seizures for 1 h. The seizures were then terminated by diazepam (30 mg/kg). Animals were killed 0, 2, 4, 8, 24 or 72 h following diazepam administration. The apoptotic and surviving neurons in hippocampus were observed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and cresyl violet staining, the expression of Smac/DIABLO, XIAP and caspase-9 was detected with immunofluorescence and western blot. The results showed that the levels of XIAP and the 46-kDa proenzyme form of caspase-9 were unaffected by the seizures. The expression of Smac increased at 2 h and the 37-kD cleaved fragment of caspase-9 was detected at 4 h, TUNEL-positive neurons appeared at 8 h and reached maximal at 24 h following seizure cessation within the ipsilateral (the same side as the intra-amygdaloid injection of kainic acid) CA3 subfield of the hippocampus. Intracerebroventricular infusion of caspase-9 inhibitor z-LEHD-fluoromethyl ketone (z-LEHD-fmk) significantly decreased TUNEL-positive neurons and increased the number of surviving cells. Caspase-9 immunoreactivity increased and Smac/DIABLO, XIAP immunoreactivity became extensive within the ipsilateral CA3 neurons. TUNEL-positive neurons and the alterations of the expression of Smac/DIABLO and XIAP within the ipsilateral CA3 were not detected within the contralateral hippocampus. These results suggest that seizures lead the translocation of Smac/DIABLO into the cytosol, the activation of caspase-9 and the change of subcellular locoalization of XIAP. These changes may play a role in the brain damage induced by seizures. Caspase-9 is possibly a potential therapeutic target in the treatment of brain injury associated with seizures.
Amygdala
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physiology
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Animals
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Caspase 9
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Caspases
;
biosynthesis
;
genetics
;
Complement Membrane Attack Complex
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Complement System Proteins
;
Glycoproteins
;
biosynthesis
;
genetics
;
Hippocampus
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metabolism
;
Kainic Acid
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Limbic System
;
Male
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Microinjections
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Protein Biosynthesis
;
Proteins
;
genetics
;
Rats
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Rats, Sprague-Dawley
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Seizures
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chemically induced
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metabolism
;
X-Linked Inhibitor of Apoptosis Protein