Table of Content

28 February 2007, Volume 27 Issue 2

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The Effects of Enrichment with Music or Colorful Light on the Welfare of Restrained Mice

HU Ying,XU Lan-wen,YANG Fei,YANG Ping

2007, 27(2):  71-76. 

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Objective To investigate the positive effects of four different forms of environmental enrichment on the welfare of mice subjected to restraint stress with the purpose of screening suitable enrichment. Method Mice undergoing poor welfare due to repeated restraint were kept in different enriched environment (enriched by march，serenade，blue filter or red filter), food consumption and body weight gain were recorded throughout the study. Two weeks later all the animals were assessed for their stress level by measuring physiological parameters (body weight, food efficiency ratio[FER], thymus index, spleen index and total leukocyte in peripheral blood[WBC]) and biochemical parameters (serum levels of ACTH，CORT，[3-EP，EPI，NE，IL-2，TP, GLU, TCHO). Result In restraint control group, restraint stress decreased their body weight gain, FER, WBC, IL-2, p-EP(P＜/i><0.01) and spleen index (P＜0.05)，while increased their ACTH,CORT, NE(P＜0.01) and EPI(P＜0.05). Compared to restraint mice, mice from blue light group and from serenade group exhibit lower levels of ACTH and NE(P＜0.01 )，as well as the restoration level of p-EP(P＜0.01) by blue light, indicating elevated neuroendocrine responses to restraint stress. With respect to the higher level of IL-2 (P＜0.01)，more WBC(P＜0.01) and larger thymus index(P＜0.01，compared to control group), blue light may contribute to relieve the inhibitory status in immune system induced by restraint. When comes to other groups, mice from red light group tended to eat more at daytime than at night compared to mice from restraint control group, and mice from march reduced their serum levels of CORT too much. Conclusion Enrichment with either serenade or blue filter partly enhanced resistance to repeated restraint stress in mice, thus suggest that both forms of enrichment should be beneficial for the welfare of laboratory mice. However, the effects of march and red light are debatable and need more investigation.

Experimental Study on Rabbit Dilated Cardiomyopathic Model

JIN Bo1,SHI Hai-ming1,ZHU Jun1,LUO Xin-ping1,HUANG Cheng-lei1,LIU Xing-dang2,WANG Yi-ping3,LIU Cong-jin2

2007, 27(2):  77-80. 

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Objective To establish Adriamycin-induced dilated cardiomyopathic(DCM) model through ear-border vein in New Zealand rabbit, and the model were evaluted by light microscopy,electron microscopy,myocardial perfusion imaging and indexes of hemodynamics. Methods Twenty male New Zealand rabbits were randomly divided into model group and control group equally. Adriamycin was injected to produce DCM model through ear-border vein. Indexes of hemodynamics were measured to evalute the cardiac function.^99mTc-MIBI myocardial perfusion imaging showed the difference between control group and model group.Histomorphological changes were evaluted by light microscopy and ultrastructure characteristics were investigated by transmission electron microscopy. Results Cardiac examinations indicated there was statistical significance compared with control group. Myocardial perfusion imaging indicated low infusion in cardiac apex, ventricular cavity became enlarged and the wall was thin.The histomophological and ultrastructure changes of myocardium coincide with the characteristics of DCM. Conclusions The adriamycin-induced cardiomyopathic rabbit model was similar to the histo-pathological changes of DCM in humans,it may be used for stem cell basic research in cardiology.

Effect of Two Anesthetics on Activity of Hippocampal Neurons in Mice

WU Dan,XIE Kun,LIN Long-nian

2007, 27(2):  81-85. 

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Objective To compare the effects of Pentobarbital sodium and Urethane on neural discharge of hippocampal CA1 region in mice, and to investigate the changes of field potentials and neural firing rates of hippocampal CA1 region after injected two anesthetic agents respectively. Methods Five male C57BL mice were used in this study. 12 tetrodes were implanted separately on the both sides of hippocampal CA1 region. Local field potentials and the neural firing activities were recorded by using the multi-channel recording technology. Results It showed the changes of field potentials and neural firing rates in hippocampal CA1 region under anesthetization of Pentobarbital sodium and Urethane respectively. Conclusions There are different effects of two anesthetics on hippocampal neural discharge and field potentials, suggesting that Pentobarbital sodium and Urethane have different mechanisms underlying neural firing activities.

Age-related Changs of Motor Function,Spontaneous Activity and Cognitive Function in C57BL/6N Mice

CHENG Yan-bo1,QIAN Jin-jun1,LIU Chun-feng1,CHENG Yan-xin2,LI Hou-da2

2007, 27(2):  86-89. 

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Objective To investigate the age-related changes in motor function’spontaneous activity and cognitive function of C57BL/6N mice. Methods Different aged, include 3,6,12,16 and 20 months old, groups of mice was constituted. Both rolling-bar test and pole-climbing test were performed to assess motor function. A computer-aided device was applied to rating spontaneous activity, and step-down test to evaluated the passive avoidance memory. Results 6 months old group of mice have significant longer rolling bar test latency than those of other groups (P＜0.05) and also manifest a shortest pole-climbing time (P＜0.05). 6，12 and 16 months old groups show a significant more spontaneous activity than those of other groups (P＜0.01). There is an age related decline in the performance of step-down passive avoidance test. Conclusion The age-related changes of motor function, spontaneous activity and cognitive function of C57BL mice have different patterns during ageing.

Effect of Different Vitrifications of Rabbits Morula Cryopreservation

CHAI Shi-qing1,2,LIU Li-jun2,YU Li-li2,CHAI Jia-qian1,XU Ping2

2007, 27(2):  90-93. 

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Objective EFS40、EFS20、mEFS40、mEFS20 solution was used to cryopreserved rabbits，morula economically and effectively. Methods Three protocols were selected in this experiment. Using mEFS40 medium ( modified EFS40) (40% Ethylene glycol+ 180ppm Ficoll70(W/v)+ 0.3mol/L Sucrose in PB1) as a cryoprotective solution，rabbit morula were equilibrated two minutes in mEFS20 medium, then equilibrated in mEFS40 cryoprotective solution one minutes. After that they were transferred into plastic insemination straws. The straws were kept on the liquid nitrogen for two minutes. Finally the straws with embiyos was put into the liquid nitrogen. The solutions used in project two were replaced with EFS20 and EFS40, the other procedure was the same as the project one. Only EFS40 solution was used in project three, the embryos were washed three times with EFS40 and transferred into the plastic insemination straws, then put the straws into the liquid nitrogen directly. All cryopreserved embryos were thawed in 25 °C water, and incubate in PB1 solution. Results The post-thaw survival in three protocols was 17.1%, 69.6% and 36.1% respectively. Conclusions The rabbits morula can be effectively cryopreserved by using the EFS40 as the cryoprotectant and EFS20 as the equilibrated solution (progeet two).

Study on Semen Parameters and Maturation of Sperm Nucleus in Male Fishar344 Rats with Spontaneous Infertility

WANG Yi-sen1,ZHU Ping-mei1,XU ping2,SHI En2,PAN Hua1,ZHOU Guang-xing1

2007, 27(2):  94-98. 

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Objective To explore the relation between semen parameters and maturation of sperm nucleus and spontaneous infertility in male Fisher344 rats. Methods Thirteen months old male Fisher344 rats (group A4) were studied comparatively with other group rats on semen parameters and sperma to genic cells and maturation of sperm nucleus. The other group rats were 4，7，10 months old male Fisher344 rats (group A₁, A₂, A₃) and 4，7，10，13 months old male S prague-Dawley rats (group B₁, B₂, B₃, B₄) and Wistar rats (group C₁, C₂,C₃,Ca). Results The percentage of sperm viability of group A4 was lower significantly than that of the other groups (P＜0.01) except group Ai (P≥0.05).The percentage of abnormal morphological sperm of group A4 was more significantly than that of the others (P＜0.01 or P＜0.05).The percentage of “a” and “b” grade motility of sperm was lover significantly than that of the other groups (P＜0.01). The percentage of Aniline blue strongly positive in sperm nucleus of group A4 was more significantly than that of the others (P＜0.01). The percentage of Aniline blue negative in sperm nucleus of group A4 was lower significantly than that of the others (P＜0.01). Conclusions Spontaneous infertility or decrease of fertility in male Fisher344 rats maybe due to the lower sperm viability, motility and maturation of sperm nucleus，and more percentage of abnormal morphological sperm.

Studies on In Vitro Fertilization by Using IVF-20 Media in Four Strains of Rat

ZHANG Chun-yan, LIU Li-jun, DI Min, ZHAO Dan-feng, ZHAO Li-hu, XU Ping, RUI Rong

2007, 27(2):  99-103. 

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Objective To investigate the feasibility of IVF-20 medium on the in vitro fertilization (IVF) in Rats. Methods Using IVF-20 as capacitation and fertilization medium, the IVF in four strains, including SD, Wistar, GK and F344 were conducted. Besides, the embryos from SD rat fertilized both in vitro and in vivo were cultured using mRlEMC medium to test their viability. Result The fertilization rates in SD, Wistar, GK and F344 were 83.2%、72.6%、87.8 % and 71.6% respectively. The rate of embiyos developed to blastula was 16.7%, having comparative viability of embryos fertilized in vivo (24.5%). Conclusion The IVF-20 medium, which had been used in vitro fertilization of human, is also a suitable medium for the capacitation and fertilization in rats.

Effect of Combining Angiostatin Gene Therapy and 5-FU Chemotherapy on Cachexia in Nude Mice Bearing with Colorectal Carcinoma in Abdominal Cavity

QIAO Wei-wei1,GU Jian-zhong2,DU Wei-dong3

2007, 27(2):  104-106. 

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Objective To induce renal transplantation tolerance by CTLA4-Ig adenovirus gene therapy in allogeneic rats and study the relative mechanisms of transplantation tolerance.Methods The kidney of SD rats were transfected with CTLA4-Ig gene-recombined adenovirus.After the orthotopic renal transplantation between SD and Wistar rats was conducted,all the rats were divided into 3 groups:group I was the control;rats in group II were perfused with empty EGFP and there in group Ⅲ with CTLA4-Ig adenovirus. The allograft survival time and the allograft funtion between the three groups were compared. Result The survival time was significantly longer in rat of group Ⅲ than in other 2 groups(31.3 ±6.8 days vs 8.2 士 1.2 days and 8.0 士 1.6 days). The serum creatinine of transfection group at 7th and 14th postoperative days was lower than that of controls at 7th postoperative day. Conclusion Local transfection of CTLA4-Ig gene can augment the tolerance and prolong the survival time of renal allograft significantly.

Effect of Long-distance Transportation on Cardiac Performance in Sub-health Beagle Dogs

PAN Yong-ming,CHEN Fang-ming,CHEN Liang,XU Jian-qin,PENG Ding-guo,ZHANG Li-zhong,CHEN Min-li,YING Hua-zhong,L(U) Jian-min

2007, 27(2):  107-111. 

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Objective Explore the effect of long-distance transportation on cardiac performance in sub-health Beagle dogs to provide a reference for the assessment of sub-health status. Methods Twelve Beagle dogs were selected in this study，six of them were fatigued by long-distance transportation, and another six as control. Blood routine，liver and kidney functions，myocardium enzyme indexes, blood sugar, electrocardiograms indexes were tested one day later. The long-distance transportation of fatigue group were confirmed as sub-healthy according to the SO2 index and clinical sub-healthy diagnostic criteria，and the cardiac performance were monitored by cardiac catheter technology. Results The average arterial pressure (MAP) and the average systolic blood pressure (SAP) in sub-healthy group are markedly lower compared with the healthy group，and the cardiac performance indexes left ventricle systolic pressure (LVSP), the left ventricle pressure (LVP), P@dp/dt_{max, +dp/dtmax, -dp/dtmax，the muscle fibres shorten rate (Vce-40，Vce(p@dp/dtmax), Vpm) significantly decreased(P-20) decreased，but the difference is not significant (P>0.05) .The total area of mental efforts loop (At)，the area of mental efforts during the contraction period (As) and the diastolic period (Ad) are markedly lowered (P0.05). Conclusion The contraction of the heart and diastole functions showed disorder in a sub-healthy state because of lower capacity to resist stress, and thus contraction in the heart, dilate during martial function can not be effectively adjusted. Therefore the use of cardiac function indexes to assess sub-health state has certain reliability and act as a reference in the clinical diagnosis and pharmacological studies.}

Comparative study on Mouse L1210 Leukemia Model Established by Different Ways

HE Hong-xing1,WU Zheng-cai1,ZHANG Wang-ping1,CHEN Cai-ping2

2007, 27(2):  112-115. 

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Objective To investigate biologic characters of a mouse leukemia model prepared by different ways. Methods The mouse acute lymphoblastic leukemia cell line L1210 was conventionally cultured and modulated into 1 xl07/ml. 0.1ml cell suspension was infused into every DBA/2 mouse by vena caudalis (A group), sub cutis (B group) and abdominal cavity (C group) respectively. The formation of leukemia in mouse was investigated. The amount of peripheric leucocyte in mouse blood was counted and the morphological changes were detected timely. The liver, spleen, kidney and lung of the dying mouse were kept，weighed and maken into pathological sections. Results The ratio of tumor formation was 100% in all groups, which spent 14，7 and 14 days respectively. The average survival days of the three groups were 24.4士3.05，24.8士1.79 and 18.8±1,79d days. The amount of peripheric leucocyte increased remarkably after infusion, which was (18.45±1.84)xl0⁹/L、(22.0±4.11) × 10⁹/L and (28.48±6.74)×l0⁹/Ltwo weeks later and (22.31±12.08)×l0⁹/L (A group) and (68.18±2.82)× 10⁹/L (B group) three weeks later. Leukemic cells could be found on peripheral blood smears. In pathological sections of the dying mouse’ liver and spleen, infused leukemic cell infiltration, mainly immature lymphocytes, could be found, accompanied with destroy of normal tissue and structure. A small quantity of leukemic cells were found in lung and kidney, destroy of their normal tissue and structure was not obviously and hemorrhage appeared in lung obviously. Conclusions Inoculation of LI 210 cells into DBA/2 mouse by vena caudalis，subcutis and abdominal cavity was feasible to prepare a mouse leukemia model. There were different biological characters, merits and defects in the model due to the different infusion ways. A certain infusion way should be selected according to the aims of the research.

Effects of Cyclophosphamide on Leukocyte of C.parapsilosis Infected Mice

LIU Xiang-mei,ZHANG Yu,MIN Fan-gui,ZHAO Wei-bo,LIU Zhong-hua,ZHANG Li-xin,HUANG Ren

2007, 27(2):  116-118. 

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Objective To investigate the effect of Cyclophosphamide on the white blood cell of C. parapsilosis infectted mouse. Methods ICR mouse were given cyclophosphamide once a day for 3 days before C.parapsilosis inoculation .The other mouse were injected with C.parapsilosis, cyclophosphamide and N.S respectively as control groups. Then the mouse blood was collected and the blood were analysised. Results After C.parapsilosis inoculation, Cyclophosphamide treatment resulted in an immediate depression of peripheral blood leucocytes, with WBC and lymphocyte rebounding quickly to normal and rising throughtout the 2-week observation period, while mouse not treated with cyclophosphamide did not. And the cyclophosphamide model mouse, the number of WBC down quickly after inoculation， while up on the 5 day and recover to normal on the 11 day. Conclusion The WBC can be depressed effectively in some times with Cyclophosphamide; Cyclophosphamide can result in immunosuppression and play an important part in C.parapsilosis murine model.

Systematic Evaluation on Biocompatibility of Coronary Stents

FANG Jing-yi,LIU Qing-fang

2007, 27(2):  119-122. 

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Objective To evaluate the biocompatibility which include blood、muscle and genotoxicity of coronary stents in rabbits. Methods Cased on ISO 10993-10，the assessments of biocompatibility were identified by experiment of the blood compatibility, genetic toxicity test and local reactions of muscle embedding test. Results 1. Coronary stents had no effects on the coagulation system of the rabbits 2. The histopathology and clinical anatomical observation of the local muscle were normal and no chromosome aberration . 3.The package remained sterile and no leakage were found. Conclusion 1. In rabbits coronary stents had no effects on the coagulation system，no local irritative reaction and genotoxicity as well. 2.The package remained no leakage and the efficacy of the package in blockage of bacteria appeared to be very effective.

Acute Toxicity and Safety Assessment of DDTs to White Cloud Mountain Minnow Tanichthys albonubes larval stages

YANG Zhi-cong,YAO Jing,FANG Zhan-qiang

2007, 27(2):  123-126. 

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Objective To study the therapeutic effect of combining angiostatin gene therapy with peritoneal chemotherapy with 5-FU on cachexia in nude mice bearing with colorectal carcinoma metastasis in abdominal cavity. Methods Angiostatin gene was cloned into adenovirus with method of RT-PCR. LoVo cell suspension was i.p to 40 nude mice. The mice were treated with combining angiostatin gene therapy and 5-FU peritoneal chemotherapy or others. Observation were made on the improvement of physiological conditions of the cachetic nude mice after treatment, and the serum levels of TNF-a and IL-1 were assayed by ELISA method. Result Compared with the controlled groups, the group of combining therapy was less loss of body mass, higher food intake, better mental condition and lower levels of TNF-a and IL-1. Conclusion Combining angiostatin gene therapy and 5-FU peritoneal chemotherapy has beneficial effect on cachexia in nude mice bearing with colorectal carcinoma in abdominal cavity.

Research Progress on HSPs Expression in Ischemic Gerbil Brain

HANG Qin,WANG Ju,DU Xiao-yan

2007, 27(2):  145-148. 

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Various studies have demonstrated an increase in heat shock protein 70 (HSP70) synthesis in the ischemic brain, suggesting a protective role for HSPs against ischemic insult. The HSPs family includes HSP90, HSP70, HSP60, HSP32, HSP27 and ubiquitin. Research progress on theses proteins in ischemic gerbil brain will be reviewed in this article.