Dynamic changes of glutamate during cerebral ischemia in the cortex of cynomolgus monkeys / 第三军医大学学报

Objective To explore the dynamic changes of glutamate in the cortex of cynomolgus monkeys during cerebral ischemia.Methods Proximal M1 segment of middle cerebral artery (MCA) was occluded for 1 h in 3 young cynomolgus monkeys (7.3 ± 1.5 years old) to induce cerebral ischemia.Magnetic resonance imaging and neurologic deficit scoring were used to evaluate the ischemia and observe the manifestations,respectively.Fast Analytical Sensing Technology (FAST) was applied to record the content of cortex glutamate in the same site of ipsilateral primary motor cortex in the periods of pre-,during,and post-occlusion,and at 1 and 2 weeks after surgery.Results Compared with pre-occlusion,the content of glutamate was increased significantly in the process of occluding in the MCA M1 (P =0.003);No significant difference was observed in the content during occluding and post-occlusion (P--0.877).The content in the first week was decreased obviously as compared with post-occlusion (P--0.004),but it showed no statistical difference with that in the second week (P =0.085).Conclusion Cerebral ischemia may potentially accelerate the extra-cellular glutamate release in the cortex,but reperfusion may ameliorate or balance off the glutamate release.

Spontaneous hypertension in cynomolgus monkeys and an analysis of its risk factors / 中国比较医学杂志

Objective To establish reference values for blood pressure in cynomolgus monkeys in different ages.Methods The blood pressures and blood lipids indexes were detected in 521 cynomolgus monkeys using an American BECKMAN-CX4 automatic biochemical analyzer and a wrist electronic blood pressure monitor.Statistical tests were performed to analyze the data.Results Significant differences were found in blood pressure values of cynomolgus monkeys in different ages.Blood pressure values in the elderly group were higher than those of other groups.The morbidity of hypertension in the elderly group was higher than those of the other groups.Body mass index (BMI) in the hypertension group was higher than that of normal group in the same age.The incidence of hypertension in the elderly group with hyperlipemia was higher than that of other groups.Logistic regression analysis showed that age, BMI and hyperlipidemia in the hypertensive group were 1.435, 1.218, and 2.337 times higher than those of the normal group when predicting the risk of hypertension.Conclusions We have initially established reference values of blood pressure in cynomolgus monkeys in different ages.Age, BMI and hyperlipidemia are risk factors of spontaneous hypertension in cynomolgus monkeys, and the measurement of blood pressure may provide a basis for the screening of cynomolgus monkey model of spontaneous hypertensive and related research.

Cynomolgus monkeys (Macaca fascicularis) experimentally infected with B19V and hepatitis A virus: no evidence of the co-infection as a cause of acute liver failure

This study was conducted to analyse the course and the outcome of the liver disease in the co-infected animals in order to evaluate a possible synergic effect of human parvovirus B19 (B19V) and hepatitis A virus (HAV) co-infection. Nine adult cynomolgus monkeys were inoculated with serum obtained from a fatal case of B19V infection and/or a faecal suspension of acute HAV. The presence of specific antibodies to HAV and B19V, liver enzyme levels, viraemia, haematological changes, and necroinflammatory liver lesions were used for monitoring the infections. Seroconversion was confirmed in all infected groups. A similar pattern of B19V infection to human disease was observed, which was characterised by high and persistent viraemia in association with reticulocytopenia and mild to moderate anaemia during the period of investigation (59 days). Additionally, the intranuclear inclusion bodies were observed in pro-erythroblast cell from an infected cynomolgus and B19V Ag in hepatocytes. The erythroid hypoplasia and decrease in lymphocyte counts were more evident in the co-infected group. The present results demonstrated, for the first time, the susceptibility of cynomolgus to B19V infection, but it did not show a worsening of liver histopathology in the co-infected group.

Establishment of a pharmacokinetic detection method for PEGylated recombinant human interleukin-11 mutein in cynomolgus monkeys / 国际药学研究杂志

Objective To evaluate the pharmacokinetics, drug concentration and effect relationship of PEGylated IL-11 mutein (PEG-mIL11) in cynomolgus monkeys through the validated anti-PEG-ELISA method. Methods PEG-mIL11 at 350 μg/kg was subcutaneously injected in cynomolgus monkeys, and the blood samples were collected at various time points. An anti-PEG-ELISA method was validated and used to investigate the concentration of PEG-mIL11, and platelet counts were measured to explore the relationship of drug concentration and effect. Results Results of the validation test demonstrated that PEG-mIL11 in monkey blood could be quantitated by anti-PEG-ELISA. Its linear range was (26.34-200) ng/ml. The specificity, accuracy and precision of the method met the present criteria. The terminal elimination half-life (T1/2) of PEG-mIL11 was (13.4 ± 2.4) h, the peak time (Tmax) was (6.7 ± 2.3) h, the peak concentration (Cmax) was (2.4 ± 0.5) μg/ml, the area under curve (AUC)(0-t) was (77.7 ± 15.6) μg∙h/ml, and the clearance (CL) was (4.6 ± 0.8) ml/ (h·kg). The thrombopoietic effect did not relate directly with the concentration of PEG-mIL11 in serum. Conclusion Anti-PEG-ELISA, used in this study to measure the concentration of PEG-mIL11, is a steady, reliable and specific method for PEGmIL11 pharmacokinetic study, and its chemical modification by PEG possesses long circulating half-lives, thereby suggesting less frequency of administration.

Experimental autoimmune encephalomyelitis in cynomolgus monkeys

Experimental autoimmune encephalomyelitis was induced in macaques. T cell clones infiltrated into the brain lesion area were compared with those in blood. Intradermal immunization of macaques with brain white matter derived from healthy macaque in combination with pertussis toxin, induced neurological symptoms in two macaques. One died on day 25 after immunization, whereas the other survived. Gross examination of the brain from the dead macaque, showed clear hemorrhagic lesions in the white matter. Hematological analysis showed that drastic T cell response was induced in macaques immunized with white matter, but not in control macaques. Flow cytometric analysis of blood cells from the affected macaques demonstrated an increase of CD4 and CD8 T cell populations expressing the CD69 early activation marker. Single strand conformation polymorphism (SSCP) analysis of T cell receptor beta chain showed T cell clones infiltrated into the brain lesion, which were different from those found in the peripheral blood of the same monkey. The present paper shows that SSCP analysis of TCR is useful in studying clonality of T cells infiltrating into the brain tissue of macaque with EAE.

Removal of human B-like antigen on cynomolgus monkey RBC by ?-galactosidase treatment / 中国输血杂志

Objective To observe the effects of enzymolysis of human B like antigen by ? galactosidase treatment on the morphology and function of cynomolgus monkey red blood cell (RBC) and to evaluate the safety of the enzyme treated RBC in animal transfusion. Methods RBC with human B like antigen was treated by recombined ? galactosidase and the effect was evaluated by absorption elution test. Meanwhile, morphology and function of the treated RBC were examined and compared with pre treatment RBC. ? galactosidase treated RBC was labeled with FITC and then infused to blood group A cynomolgus monkeys. Flow cytometry was used to detect the survival of donor RBC in recipient’s blood. Blood chemistry and urinalysis of recipients were performed before and after transfusion. Results The human B like antigen of cynomolgus monkey RBC was obliterated by ? galactosidase treatment and the treated RBC maintained normal morphology and function. Survival at 24 h after transfusion was 84.6% and 68.1%. T 1/2 were 7d and 8d versus 13d in the controls. There were no change in blood chemistry and urinalysis of the recipients. Conclusion Enzymolysis of cynomolgus monkey RBC does not affect the cellular function and morphology. Transfusion of the enzyme treated human B like RBC into human A like cynomolgus monkeys is safe.

Research on demyelinating mechanism in acute experimental allergic encephalomyelitis in cynomolgus monkeys / 中国免疫学杂志

Objective:To study demyelinating mechanism of the central nervous system in acute experimental allergic encephalomyelitis(EAE).Methods:EAE in cynomolgus monkeys was induced successfully by homologous brain white matter homogenate. Lymphocyte subset in blood and cerebrospinal fluid was monitored by flow cytometer. Pathological changes in brains of acute EAE monkeys were investigated by immunohistochemistry and electron microscopy. Results:In the CSF of acute EAE, CD4 + lymphocytes increased significantly, and CD8 + lymphocytes and B lymphocytes increased slightly whereas the control was normal. A lot of CD4 + lymphocytes and a few CD8 + lymphocytes infiltrated into temporal deep white matter of acute EAE, whereas the control was normal. Inner laminae of myelin sheath were loose and axon was separated, whereas outer laminae were normal. Although axon was preserved well, oligodendrocyte had a severe edema in cytoplasm with mitochondria swollen, crista blurred or broken, and nucleus lysised partly.Conclusion:It is oligodendrocyte rather than myelin sheath itself which is firstly attacked in the demyelination in EAE.