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Isolating technologies development and used in gnotobiology

Chinese Journal of Comparative Medicine.2014;(5):76-79. doi:10.3969.j.issn.1671.7856.2014.005.016

To introduce germfree isolating technologies used in gnotobiology , and to compare the differences with in Cleaning Rooms, Restrictive Access Barrier Systems (RABS) and Isolators.To expect the development of isolators , and to describe the uses of isolators in gnotobioly .

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The study on organ weights, blood physiological and biochemical parameters and immune cells in NOD/SCID mice

Huaiyuan WANG ; Xuying WU ; Wenming ZHANG ; Yushu TIAN ; Jinfeng WU ; Yu WANG ; Yajing ZHANG ; Zhiwei YANG

Chinese Journal of Comparative Medicine.2014;(5):47-51. doi:10.3969.j.issn.1671.7856.2014.005.010

Objective To measure the organ weights , physiological and biochemical parameters and immune cells of NOD/SCID mice.Methods Mice at five and ten weeks of age were selected .The organ weights, blood physiological and biochemical parameters were observed .The percentages of CD 3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were checked by FCM in NOD/SCID mice at six week of age in terms of its T , B lymphocyte function and NK cell activity.Results Among the same sex group of NOD/SCID mice, the weights of kidney, liver, heart and lung, and the blood physiological indexes of RBC , HGB, HCT, MCV, MCH, RDW, and the blood biochemical indexes of TP , ALB, ALP, CHO, TBIL show significant difference between 5 and 10 weeks.At the same age, HCT, GLOB, A/G, CHO, TG, TBIL and the UN are significantly different between male and female NOD /SCID mice.NOD/SCID mice lack T cells (0.37 ±0.26)%、CD4+T cells (0.35 ±0.13)%、CD8 +T cells (0.47 ±0.10)%、CD19+B cells (0.13 ± 0.05)%、B220+B cells (1.20 ±0.44)%.The percentage of NK cells is (6.90 ±0.82)%, and the percentage of granulocytes is (47.88 ±15.54)%.Conclusions The study indicates that NOD/SCID mice show the deficiency of T , B and NK cells function .The organ weights , blood physiological and biochemical parameters are affected by age and gender . The studied parameters of NOD/SCID mice are similar with the same strain in other countries .

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Observation and analysis of the individual behavior with weaning of infant Rhesus Monkeys

Yanyan LI ; Lixiong CHEN ; Sudong QI ; Junbin WANG ; Fengmei YANG ; Yuan ZHAO ; Shuaiyao LU ; Wenhai YU ; Zhanlong HE

Chinese Journal of Comparative Medicine.2014;(5):35-41. doi:10.3969.j.issn.1671.7856.2014.005.008

Objective To understand the characteristics and rules of behavior and preliminary study on the change of psychological state and behavior of infant rhesus monkeys after weaning by artificial feed .Methods 290 infant rhesus monkeys were born in 2012, individual behavior of infant rhesus monkeys were observed and recorded by the methods of ad libitum sampling and chronography .Results The behavior of huddling , sucking finger and sucking body of companion were observed in the first month , the behavior of huddling and sucking finger were significantly increased in the second and third month(P <0.05), sucking body of companion in second month were significantly higher than the first month (P <0.05);the new behavior pacing , sucking genitalia and mounting were observed in the second month , pacing and sucking were significantly increased in the third month ( P <0.05 ) , mounting did not have obvious change; in the third month , sucking toe behavior ware recorded and did not change with time; The proportion of huddling behavior occurred between female and female was significantly higher than opposite sex or male in the same cage .The male infant monkeys were more likely to occurred pacing behavior than female .Sucking body of companion and sucking finger behavior of male were significantly lower than the female infant monkeys ( P <0.05 ) .There was no significant difference of sucking toe behavior .Conclusions 7 behaviors were recorded during the whole stage of observation , huddling behavior , pacing behavior , sucking toe behavior , sucking finger behavior , sucking genitalia behavior , sucking body of companion behavior and mounting behavior respectively , and 7 behaviors were increased by degrees with time .Huddling behavior to reflect fear psychology;sucking behavior to represent physiological demand and defensive psychology ;pacing was insignificant behavior in the initial stage , then to reflect psychological anxiety of rhesus monkeys .Provided a way and model on assessment of psychology, praxiology, to study and research the change of psychological state and behavior of early weaning infants .

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Effects of mild hypothermia induced by pentobarbital sodium on hematology in male BALB/C mice

Shufang LI ; Liqun HUANG ; Juan YUE ; Yongli WANG ; Xingang WANG ; Weibin LI ; Quan AN

Chinese Journal of Comparative Medicine.2014;(5):31-34. doi:10.3969.j.issn.1671.7856.2014.005.007

Objective To observe the effects of mild hypothermia induced by pentobarbital sodium on hematology in male BALB/C mice.Method Healthy male BALB/C mice were divided randomly into two groups:control group ( C) and mild hypothermia group(M).The body temperature of the mild hypothermia group was maintained between 28℃ to 30℃( anal temperature ) for 4 hours induced by pentobarbital sodium injected intraperitoneally , then recover unaffected . Anal temperature, coagulation, electrolytes, and blood cell indexes were examined in 2, 24, 72 hours after treated by mild hypothermia;Control group was given equal volume of saline volume at constant temperature .Results The body temperature and coagulation in mild hypothermia group showed no significant difference compared with the control group ( P﹥0.05),but the concentration of K +and Na +in mild hypothermia group were higher than control group (P﹤0.01), the number of WBC in mild hypothermia group was lower than control group ( P﹤0.01或P﹤0.05 ) , and the RBC、HGB、MCH、MCHC in mild hypothermia group were lower than control group transiently (P﹤0.01或P﹤0.05).Conclusion Mild hypothermia induced by pentobarbital sodium affects some of hematological values in mice considerably .

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Gene silencing of 90K/Mac-2BP enhances the apoptosis of U937 cells by HIV-1 infection

Chunyan FU ; Hong JIANG ; Jing XUE ; Zhe CONG ; Ting CHEN ; Qiang WEI

Chinese Journal of Comparative Medicine.2014;(5):10-14. doi:10.3969.j.issn.1671.7856.2014.005.003

Objective To investigate the effect of cell death by HIV-1 infection on gene 90K/Mac-2BP by RNA interference (RNAi) in U937 cell line.Methods We used human monocyte-macrophage cell line U937 as the cell model.Cells were infected by HIV-1 ( R5-tropic) 5 days, and then stained by PE-Annexin V and PerCP-7-AAD.90K/Mac-2BP in U937 cell line was knocked down , and these cells were infected by HIV-1 for 5 days.Then, cells were stained by PE-AnnexinV and PerCP-7-AAD.Apoptosis were examined upon flow cytometry .Results The percentages of Annexin V+cells without 90K/Mac-2BP knock-down were (16.27 ±0.30)% by HIV-1 infection.The percentages of them with 90K/Mac-2BP knock-down were (31.26 ±0.35)%, (25.76 ±0.30)%, (23.69 ±0.33)% respectively.The increase of cell apoptosis rate for HIV-1-infected U937 cells by 90K/Mac-2BP siRNA transfection was significantly greater than that for HIV-1-infected untreated cells (P﹤0.01).Conclusion The apoptosis of HIV-1-infected U937 cells was regulated by the expression of 90K/Mac-2BP.

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Effects of rhodiola crenulata compound on serum corticosterone and myocardial glucocorticoid receptor in rats exposed high sustained positive acceleration

Liangen CHEN ; Hao ZHAN ; Feng WU ; Andong ZHAO ; Minghao YANG

Chinese Journal of Comparative Medicine.2014;(5):1-4. doi:10.3969.j.issn.1671.7856.2014.005.001

Objective To study the effect of rhodiola crenulata compound on serum corticosterone and myocardial glucocorticoid receptor ( GR) in rats exposed high sustained +Gz.Methods Seventy-two healthy SD rats were randomly divided into six groups: blank control group, stress control group, +10 Gz stress group and low, medium, high dose drug group, with twelve rats in each.20 mL/kg menstruum was fed to each rat once per day for 14 days.Low, medium and high dose drug group were fed with rhodiola crenulata compound at doses of 0.75 g/kg, 1.5 g/kg and 3.0 g/kg respectively, the other three groups were fed with equal volume saline.Rats were exposed to high +Gz in 15th day.The concentration of corticosterone in the serum of each group rats was detected with ELISA. Western blot was used to detect the expressions of GR in the myocardium of each group rats .Results The content of corticosterone was significantly elevated in +10Gz stress group, while the expression of GR in the myocardium was markedly declined (P <0.01,P <0.05).However, compound preconditioning could decrease concentration of corticosterone in the serum and enhance the expression of GR in the myocardium from rats after +10 Gz exposure. The corticosterone concentration of medium and high dose drug groups was significantly lower and the level of GR expression in the myocardium was significantly higher than that of +10 Gz stress group ( P <0.05 , P <0.01 ) . Conclusion Rhodiola crenulata compound preconditioning could regulate the concentration of corticosterone in the serum and the level of GR expression in the myocardium of the rats exposed +10 Gz, which may be related with its protective effect on high sustained +Gz-induced injury of myocardium.

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The construction and use of the operating room for in vitro fertilization and embryo transfer (IVF-ET) in non-human primates

Qinfang JIANG ; Jiahong GAO ; Qiao ZENG ; Donghong TANG ; Zhanlong HE ; Kaili MA

Chinese Journal of Comparative Medicine.2014;(5):72-75. doi:10.3969.j.issn.1671.7856.2014.005.015

The non-human primates have been profoundly being used to study the human reproductive models .It is the prerequisites to set up the operating rooms including hardware and software on reproductive experiments and transgenic manipulation.Thus, Combined with the practice of our center , both the construction and the associated problems about the IVF-ET operating rooms were discussed in this paper .It will provide some help for the related researches .

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Development and application of computer-auto-controlling and analysis system for tail suspension

Xiuping SUN ; Xiaomeng ZHANG ; Cong LU ; Yihua LI ; Xinmin LIU

Chinese Journal of Comparative Medicine.2014;(5):66-71. doi:10.3969.j.issn.1671.7856.2014.005.014

Objective To develop a computer-auto-controlling and analysis system for Tail suspension .Methods Combining the advantage of computer science , engineering and animal behavior into tail suspension test .The “energy”index was developed .The system was validated with antidepressants such as imipramine and paroxetine .Results The measuring principle is based on the energy developed by mice trying to escape from their suspension .During the test, the movements of the mice are analyzed in terms of force , energy and power developed over time .Each mouse issuspended by the tail using adhesive tape to a hook connected to a tail test sensor .The tail test sensor fixed to suspension bar picks up all movements of the mouse and transmits these to a signal regulation unit and transmission circuit , which amplify , filter and digitalizes the signals.The signals are displayed visually in waveform .Activity time, immobility time, energy are continuously updated .The correlation coefficient of “immobility time” collected by computer and manpower was 0.94. Using this system, both imipramine and paroxetine could decreased the immobility time and paroxetine could increased the energy induced by mice ( both P <0.01 ) .Conclusion A stable computer-auto-controlling and analysis system for Tail suspension was established and could be used to screen the antidepressants .

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Chronic hepatic injury modeling in mice induced by D-galactosamine and lipopolysaccharide combination

Yanan ZHAI ; Jingjing WANG ; Meng LI ; Yafei CHI ; Xia MENG ; Boya ENG ; Kun JIAO ; Jing LU

Chinese Journal of Comparative Medicine.2014;(5):62-65. doi:10.3969.j.issn.1671.7856.2014.005.013

Objective To research the method of Chronic hepatic injury modeling in mice induced by D -galactosamine and lipopolysaccharide combination . Methods Injected D-galactosamine ( 30 mg/mL ) and lipopolysaccharide ( 2μg/mL ) combination by intraperitoneal injection , two days at a time for 8 weeks .Monitored variation of diet and weight; detected serum level of alanine aminotransferase ( ALT ) and aspartate aminotransferase (AST), been put to death in mice and removed the liver tissue .strained hepatic tissue by the HE and Masoon dye to observe Liver tissue structure and cellular morphology and the degree of fibrosis .Results Lipopolysaccharide and D-galactosamine combination resulted in ALT rise , hepatocyte degeneration and necrosis ,collagen fiber hyperplasia obviously . Conclusion D-galactosamine and Lipopolysaccharide combination could induce mice chronic hepatic injury modeling .

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The study on organ weights, blood physiological and biochemical parameters and immune cells in Rag2 knockout mice

Yu WANG ; Xuying WU ; Zhaosheng WEI ; Jinfeng WU ; Huizhen LIU ; Yushu TIAN ; Wenming ZHANG ; Zhiwei YANG

Chinese Journal of Comparative Medicine.2014;(5):56-61. doi:10.3969.j.issn.1671.7856.2014.005.012

Objective To measure the organ weights , blood physiological and biochemical parameters , and immune function of Specific Pathogen Free ( SPF) Rag2 knockout ( KO) mice.Methods Rag2 knockout mice were selected at five and ten weeks , and the organ weights , blood physiological and biochemical parameters were observed .The percentages of CD3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were checked by FCM in Rag2 KO mice at six week of age in terms of its T , B lymphocyte function and NK cell activity .Results Among the same sex group of NOD/SCID mice, the weights of brain, lung, spleen, liver, heart, kidney, and the levels of TBIL, WBC in 10 weeks of age are higher than 5 weeks.At the same age, the weights of heart, kidney, liver, spleen, and the levels of AST, ALP, A/G, GLU, PLT, PCT, WBC, and LYM%in male mice are higher than females.The Rag2 KO mice lacks T cells(0.36 ±0.15)%、CD4+T cells(0.21 ±0.06)%、CD8+T cells(0.23 ±0.07)%、CD19+B cells(0.28 ±0.04)%、B220+B cells(2.03 ±0.42)%).The percentage of NK cells is(24.13 ±3.62)%, and the percentage of granulocytes is (57.20 ±3.85)%.Conclusion The study suggests that the organ weights , blood physiological and biochemical parameters are affected by age and gender in Rag2 KO mice, which main biological characteristics are similar with C 57BL/6J mice.The Rag2 KO mice show the deficiency of T , B cells function .

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