Table of Content

25 June 2012, Volume 32 Issue 3

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Chronic Unpredictable Mild Stressed Rats Increased Their Movements in Open Field

LI Xia, WU Cheng-ye, XIAO Shi-fu, ZHENG Xian-jie, DONG Xiao-wei, PENG Su-fang, WANG Tao, XIAO Ze-ping

2012, 32(3):  175-181.  DOI: 10.3969/j.issn.1674-5817.2012.03.001

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Objective To explore the influence of chronic unpredictable mild stress(CUMS) on rats' open field behaviors. Methods Twenty-four SD rats were randomly assigned to three groups: A is control group, B is CUMS group, C is CUMS + citalopram group. Rats in group B and C were subjected to the CUMS schedule; Group A and B were submitted to administration with saline, while group C was pretreated with citalopram (CTP, 10 mg/kg, i.p.) every day. Body weight was measured every week. Sucrose preference and open field test were evaluated every three weeks. Animals were sacrificed after 6-week CUMS trial. Results CUMS rats, either with or without citalopram, weight gain was reduced compared to control (average 153.05 g, 127.69 g, 232.57 g, respectively). Rats in CUMS group had lower sucrose preference than which in control group and CUMS + citalopram group (average 60%, 73% and 71%, respectively). However, several behavioral indicators in open field, including total distance travelled, total mobile time and number of line crossings, were increased in CUMS treated rats,while which in control and CUMS + citalopram rats declined. Conclusion Chronic unpredictable mild stress may lead to rat's depression and elicit simultaneously some anxiety-like symptoms, such as increased motor activity.

Depression in Rat Model Sucrose Preference Test Data Analysis of Aggregation

HOU Jin-liang, WEI Sheng, DU Xi-yang, ZHANG Hui-yun

2012, 32(3):  182-185.  DOI: 10.3969/j.issn.1674-5817.2012.03.002

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Objective To explore the rat model depression sucrose preference test (Sucrose preference test, SPT) in sucrose preference value SP (Sucrose preference) the best aggregation. Methods Chronic mild stress (CMS) rat model of depression was replicated. In the four weeks of modeling process, gathering a sucrose preference test data for every other week, and the sucrose preference test data using twi-two aggregation analysis approach. Aggregation of different means than the preparation of model reliability and correlation effects. Result ① Kappa TM showed only way to get a higher aggregate reliability (Kappa = 0.611), ② ICC results showed that the aggregation of TM's credibility highest (ICC = 0.910)). Conclusion The appropriate data aggregation can significantly improve depression in rats with sucrose preference test data credibility and stability.



Study on BMP-2 on Inducing OPG^-/-BMSCs Differentiate into Osteogenesis in vitro

ZHANG Chun, WEI Qin, WANG Xue-mei, KUANG Ying, DUAN Ming-jun, ZUOFEIYA·Keyimu, JIANG Tao, MA Mei, WEN Hao

2012, 32(3):  186-190.  DOI: 10.3969/j.issn.1674-5817.2012.03.003

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Objective To investigate the effect of bone morphogenetic protein-2(BMP-2) on inducing OPG^-/- BMSCs differentiate into the osteogenesis in vitro, and to evaluate the effect of BMP-2 Bioactive Peptides inducing to osteoplast and dose dependent. Methods Got the femur of mice, separated the BMSCs and cultured in vitro. Followed by passaged the BMSCs to P1 using the conditioned medium, and then the BMSCs were added in the BMP-2 consistency of 0.3 μg、0.2 μg、0.1 μg、0.05 μg、0 μg separately cultured with 15% FBS culture medium, marked A, B, C, D, and E group in order. The 3^rd, 5^th, 7^th day after cell culture, the expression of alkaline phosphatase was detected by PNPP method, and calcium content was detected by Calcium kit. The expression of tatrate-resistant acid phosphatase form 5 b in the cell supernatant detected by ELISA, To investigate the effect of different concentration bone morphogenetic protein-2 (BMP-2) on inducing OPG^-/- BMSCs differentiate into the osteogenesis in vitro. Results The activity of alkaline phosphatase and calcium content showed that group A and group B both increase higher than the other group (P<0.05). However no significant differences were detected between the group A and B (P>0.05). TRACP5b displayed, TRACP5b expression in group A, B and C is lower than the other groups in the 3^rd, 5^th, 7^th day (P<0.05), no significant differences were detected between the group A and B (P>0.05). Conclusions The BMP-2 had significant effect on inducing OPG^-/- BMSCs differentiate into the osteogenesis in vitro. The effect dependent on the different concentration of bone morphogenetic protein-2(BMP-2), the optimal concentration is 0.2 μg/ml.



Effect of Bone Marrow Stem Cells on Repairment of Ileum Radiation Damage

SONG Xiang-fu, SUN Bing-xue, ZHANG Jing-chun, GONG Shou-liang

2012, 32(3):  191-194.  DOI: 10.3969/j.issn.1674-5817.2012.03.004

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Objective To observe the migration, colonization and repairing effects of marrow mesenchymal stem cells (MSCs) on ileum damage induced by radiation in rats. Methods The model of ileum radiation damage was made. Four rats from the ileum radiation damage model were sacrificed at 2 and 7d respectively after they were injected mesenchymal stem cells 1 ml (concentration:1 × 10⁶/ml), and ileal tissue were taken out, then the enrichment of MSCs in the ileum were observed under confocal microscope. Thirty-six rats were randomly divided into normal control group, model, and MSCs group (n=12). Animals in each group after the last injection of MSCs were sacrificed at 3 and 7 d, pathological changes of the ileum was observed. Results The occurrence of MSCs was observed in the ileum 2 d after MSC infusion, and widely distributed around the mesenteric vessels 7 d later. The intestinal structure was clear in normal control group, and epithelial cell was significantly necrosis at 3 d, involving the whole layer of cell degeneration in model group; compared with the model group, mucosal get thickness, and intestinal necrosis was relieved 7 d in treatment group. Conclusion The MSCs can be rapidly enriched in ileum radiation damage and promote regeneration and repairment.



Effects of Shenqifuxin Oral Liquid on Plasma Kaliuretic Peptide, Myocardial Contractility and Relaxation of Left Ventricle in Rats after Sleep Deprivation

MA Lu, ZHU Xiao-fa, QIAN Yan-fang, LI Xiu-yu, ZHAO Mei-na

2012, 32(3):  195-198.  DOI: 10.3969/j.issn.1674-5817.2012.03.005

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Objective To observe the effects of Shenqifuxin oral liquid (SQFXOL, 参芪扶心口服液) on plasma kaliuretic peptide (KP) and the left ventricular remodeling and the myocardial contractility and relaxation of left ventricle in rats after sleep deprivation. Methods The hemodynamic parameters were measured by the method of the catheterization. Plasma concentrations of KP, ANP, AngⅡ and ET-1 were determined by radioimmunoassays. Results In high dose group, the LVSP, dp/dtmax and Vmax were increased; LVEDP was decreased, plasma concentrations of KP, AngⅡ and ET-1 were decreased significantly(P<0.05 or P<0.01). The levels of plasma ANP in middle dose group and low dose group were significantly lower than those in model group (all P<0.01). Conclusion SQFXOL can reduce the plasma concentrations of KP, AngⅡ, ET-1, and ANP, improve the myocardial contractility and relaxation of left ventricular in rats with HF after myocardial infarction.

Catheterization Caused Congestive Heart Failure Rabbit Model of Heart Qi deficiency

WANG Xiao, DONG Hao-ran, LIN Chuan-quan, XU Qing-wen

2012, 32(3):  199-202.  DOI: 10.3969/j.issn.1674-5817.2012.03.006

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Objective To construct congestive heart failure rabbit model of heart Qi deficiency caused by catheterization. Methods Construct cardiac hyper-volume overload, rabbit heart failure model was caused by catheterization.Fifteen and 30 days after modeling, the general status, feed consumption index, body weight index, heart rate, respiration and ECG were observed and measured. Results Compared with the normal control group, SBP, DBP, LVSP and other hemodynamic index decreased significantly. After 15 and 30 days of modeling, the activity of model's rabbit was significantly reduced, the apathetic, efforts of resisting grab were reduced, feed consumption and body weight indices were decreased, breathing and heart rate were increased significantly, abnormal Q wave was appeared, and ST wave segment was elevated. Conclusion The heart rate, respiration, ECG, body weight and feed consumption index and other indicators can qualitatively and quantitatively reflect the heart failure model of heart qi deficiency.



Using Laser-Punched System on IVF of Fresh and Cryopreservated and Post-Thawed Sperm

LIU Li-jun, ZHU Quan-feng, YU Li-li, ZHANG Yi-da, SHI Mei-lian

2012, 32(3):  203-208.  DOI: 10.3969/j.issn.1674-5817.2012.03.007

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Objective The IVF rate of the fresh sperm and the post-thawed sperm about C57BL/6J mice were compared, and then C57BL/6J oocytes were laser-punched, and these oocytes were IVF with fresh and post-thawed sperm, their IVF rate were investigated. Method Two inbred mice strains of(C57BL/6J, C57BL/6N) and 5 strains of genetic engineering mice (MKP-5, Sir-168, C57BL/6RLR 3-/-, C57BL/6 TRL 2KO, C57BL/6 TRL 4KO) were chosen to freeze sperms. Firstly, the IVF rate of fresh sperm and post-thawed sperm with normal C57BL/6J oocytes were compared. Secondly, C57BL/6J oocytes were laser-punched, and then the IVF rate of the fresh sperm and post-thawed sperm with these oocytes were compared. Result (1) The IVF rate of fresh sperm and post-thawed sperm of C57BL/6J, C57BL/6N were 50.4% vs 17.7%, 48.6% vs 15.4%; when the oocytes were laser-punched, the IVF rate were 64.6% vs 59.7%, 60.4% vs 15.4%; (2) the IVF rate of fresh and frozen sperm of 5 genetic engineering mice were 91.5% vs 28.6%, 77.4% vs 52.3%, 40.7% vs 12.7%, 56.5% vs 14.6%, 65% vs 6.5%, when those sperm were fertilized with punched C57BL/6J oocytes, the result were 75% vs 50.7%, 74.3% vs 35.6%, 75.7% vs 32.7%, 79.5% vs 48.9%, 59.5% vs 46.3%. Conclusion When oocytes were laser-punched, the IVFrate of post-thawed sperm were increased significantly, it is showed that laser-punched system was a useful and economical way to increase the IVF rate of mice frozen sperm.

Comparation of Dendritic Cells from Different Sources of WHBE Rabbits

TU Jue, XU Jian-qin, PAN Yong-ming, LIU Zhi-hua, KONG Qin, CHEN Min-li

2012, 32(3):  209-214.  DOI: 10.3969/j.issn.1674-5817.2012.03.008

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Objective To generate dendritic cells (DC) from different sources of WHBE rabbits (Oryctolagus cuniculusz). Methods DC were generated from mononuclear cells of bone marrow, peripheral blood and spleen by GM-CSF and IL-4. The morphology and ultrastructure were observed and the phenotypes of DC were analyzed. Immunological fuctions of the three differently derived dendritic cells were examined by allo-MLR. Results DCs generated from mononuclear cells of bone marrow, peripheral blood and spleen were matured by lipopolysaccharide (LPS). All three types of the mature DC exhibited extensive dendrites on their surfaces. DCs from peripheral blood and spleen were matured earlier than bone marrow. About 10⁶-10⁷ DCs were generated from three different sources and most of all was that from peripheral blood. All types of the DCs showed higher expression of CD86 compared with isotype control and obvious capacity to stimulate primary T cell responses in mixed lymphocyte reaction (MLR). But there were no difference among three groups (P>0.05). Conclusion Peripheral blood mononuclear cells (PBMC) were convenient and economic source to generate DC of WHBE rabbits.



Observation on Chest Big Vascular System of Rhesus Monkey by Three-dimensional Spiral CT Reconstruction

FAN Chun-mei, LI Zhi-xiong, CHE Min

2012, 32(3):  215-217.  DOI: 10.3969/j.issn.1674-5817.2012.03.009

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Objective Observing rhesus monkey big vascular system of chest by 256 layer spiral CT and providing scientific materials for studying rhesus monkey vascular and digitization system. Methods Five male and five female rhesus monkeys aging at 6 to 10 years old were selected and obtained for CT scanning experiment. Rhesus monkey chest CT cross-sectional images were acquired through flat and strengthen scanning technique to collect volume data. Rhesus monkey volume data were processed by 3D image restruction using CT image post-processing technology. Results Rhesus monkey stereoscopic images of aorta, pulmonary artery and vein and chest big vascular were obtained by Three-dimensional reconstruction. From the point of comparative anatomy the difference of chest big vascular system between rhesus monkey and human being were found. Conclusion The anatomical and tissue structures of rhesus monkey among the big vascular system were reflected perfectly by Three-dimensional CT reconstruction. It is an effective, non-invasive and direct ideal way of observation of the vascular system.



IL-12 Facilitate the Recovery of Hematopoietic Stem Cells after Ionizing Radiation in Rat

WANG Cui-ling, WANG Zeng-song, XIA Shu-qi, GUO Gui-ling, XIE Fei-qun, LI Wen-tang, NIU Li-hong, LUO Qun-huang, ZHANG Yi-jun

2012, 32(3):  218-221.  DOI: 10.3969/j.issn.1674-5817.2012.03.010

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Objective The ability of recombinant murine interleukin-12 (rmIL-12) to facilitate recovery of bone marrow hematopoietic stem cells (HSC) from irradiation in mice model were evaluated. Method The rmIL-12 treated SPF BALB/c mice or bone marrow hematopoietic stem cells obtained from the animals before and after irradiation were examined for the viable counts and apoptosis of bone marrow hematopoietic stem cells. Results The rmIL-12 treatment before and after irradiation was effective to increase hematopoietic stem cells of suppressed bone marrow and reduce apoptosis both in obtained hematopoietic stem cells and in mice. Conclusion The rmIL-12 had obvious effect on recovery of hematopoietic stem cells from irradiation induced damage. Moreover, this study provided experimental evidence for IL-12 as clinic therapy to treat diseases associated with hematopoietic cytopenia.



Analysis on Reproductive Performance and Growth of APPswe/PS1dE9 Transgenic Mice Model for Alzheimer's Disease

LI Na, WANG Wei, LI Chen-jing, ZHANG Ying-yan, ZHANG Sheng, ZHANG Hai-yan, ZHANG Zhou

2012, 32(3):  222-227.  DOI: 10.3969/j.issn.1674-5817.2012.03.011

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Objective To afford experimental basis for establishing the standard biological properties' datum of APPswe/PS1dE9 transgenic mice model on Alzheimer's disease. Methods The heterozygote of the APP/PS1 transgenic mice were used to breed, and the identify method of genotype was established. The reproductive performance and growth of different litters and genotype were analysed within four litters of 1158 mice in 160 litters. Results ① The section of APP/PS1 was 600 bp. ② The quantity of the newborn mice and transgenic positive in litter 2 and 3-were more than litter 1(P<0.05), but the positive rate had no significant difference(P>0.05). ③ The birth interval between parity 1 and 2 was shorter than other's (P<0.05). Litter size, number of mice weaned and survival rate after weaning of the parity 2 and 3 were higher than other two (P<0.05). ④ The litter weight at birth, lactescence, body weight after weaning of parity 3 and 4 were stable. ⑤ The highest weight of female and male APP/PS1 transgenic mice are 24.45±0.92 g and 30.21±2.53 g separately. ⑥ This transgenic mouse genotype did not affect its weight. Conclusions The basic biological characteristics data of APP/PS1 transgenic mouse are established, and provide the exact data for scientific research.



Development of Self-learning System for Laboratory Animal Science

HU Ying, YANG Fei, YU Yi-jun, GAO Cheng

2012, 32(3):  228-233.  DOI: 10.3969/j.issn.1674-5817.2012.03.012

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Objective To develop a self-learning software for laboratory animal science based on the technology of computer assisted learning. Methods The framework of the software was designed on the basis of animal category, meanwhile, all the points in laboratory animal science were organized through two classes of subjects, then,different kinds of information were matched and integrate into the program by using Microsoft Visual Studio 2010, Microsoft.NET Framework 4.0 and C#. Results The whole software includes an entrance with the function of selecting animal and some learning interfaces offering information about the first or second subjects, all these were established on the relationship between database and process of the program. Conclusion The self-learning system for Laboratory Animal Science developed in this study is expected to be a practical tool in self-teaching, self-training and searching information, it also provide a primary computer assisted learning environment for Laboratory Animal Science.



Application of Germ Free Animals in Study of Human Health and Disease

ZENG Ben-hua, WEI Hong

2012, 32(3):  250-253.  DOI: 10.3969/j.issn.1674-5817.2012.03.018

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Gut microbiota has been reported to involve in host physiology functions, certain pathological disorders, and drug metabolism. The gut microbiota colonization is also involved in several important development of physiological function, especially development of immune system and digestive system. As the cleanest animal associated no microorganism, germ free animal is a precondition to study the relationship between microbiota and human health state. Germ free animal, gnotobiotic animals and human flora associated animals have been widely used in studying the effect of microbiota on host development, etiological factor of metabolic disease and drug metabolism.



Progress on Mongolian Gerbil Applied in Research of Helicobacter pylori, Hearing and Anxiety

XIE Xin-yu, DU Xiao-yan

2012, 32(3):  254-258.  DOI: 10.3969/j.issn.1674-5817.2012.03.019

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Mongolian gerbil is a being developed laboratory animal, so-called “multi-function” experimental animal and has been widely applied in many research fields. Several new medical research progresses on Mongolian gerbil including hearing, helicobacter pylori, anxiety and others research fields were briefly reviewed in this article,.