Table of Content

25 April 2015, Volume 35 Issue 2

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Establishment of Vascular Restenosis Model Induced by Carotid Artery Balloon Injury in Rats

YANG Bo, WU Bing, TANG Jun-ming, YANG Jian-ye, CAO Teng, WANG Jia-ning

2015, 35(2):  87-91.  DOI: 10.3969/j.issn.1674-5817.2015.02.001

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Objective To establish a simple and useful method of rat carotid artery balloon injury, providing stable and reliable animal model for studying the changes of vessels based on physical damage. Methods Rat common carotid artery, external carotid artery and internal carotid artery were isolated after anesthesia with isoflurane, the artery incision was performed in the external carotid artery, and then the 2F balloon catheter was inserted into common carotid artery in the incision, and pushed about 4 cm. Meanwhile, 0.02 mL normal saline was slightly injected to make the balloon inflate, and then the catheter was gently rotated for all the way when removing. Repeat this operation by three times to completely denude the common carotid artery intima. The changes of histomorphology in carotid artery were observed 1 day, 3 days, 7 days, 14 days, and 28 days after injury. Results Obvious neointima hyperplasia appeared over time after damage, which reached the peak at 14 days. Conclusion The improved rat carotid artery balloon injury model is established with the characteristics of simple operation, high success rate, and low rate of thrombosis.



Protective Effects and Mechanisms of 4,4'-diisothiocyanostibibene-2,2'-disulfonic Acid Injected by Intarcerebroventricular against Cerebral Ischemia-reperfusion Injury Rat

YUAN Zheng, ZHANG Lu, WU Wen-qing, YUAN Zi-yan, ZHAO Shan-min, YU Chen-lin, LIN Li-fang, TANG Qiu

2015, 35(2):  92-96.  DOI: 10.3969/j.issn.1674-5817.2015.02.002

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Objective To explore the protective effects and mechanisms of 4,4'-diisothiocyanostibibene-2,2'-disulfonic acid (DIDS) injected by intarcerebroventricular against cerebral ischemia-reperfusion injury (CIRI) in middle cerebral artery occlusive (MCAO) rats. Methods Focal cerebral ischemia in rats was produced by 2 h occlusion of the middle cerebral artery and 24 h reperfusion. DIDS and normal saline (NS) was injected by intarcerebroventricular. The neurologic deficit scores were investigated according to Zea-Longa's Standard. The infarct areas were assessed with software Imagepro Plus 6.0 after TTC staining. The levels of Bcl-2 and Caspase-3 in central neutral system were assessed by western blot. And the activities of inducible nitric oxide synthase (iNOS) as well as glutathione peroxidase (GSH-PX) were determined by specific kit. Results DIDS (100 μmol/L) could decrease the neurologic deficit score, reduced the infarct areas of the brain in CIRI rats, enhance the expression of Bcl-2, depresse the expression of Caspase-3, and decrease the activity of iNOS. Conclusion DIDS has a protective effect on CIRI of rat and this effect is related to enhancing the expression of Bcl-2 and diminishing Caspase-3 expression, decreasing the activity of iNOS.



Effect of Induced Obesity on Metabolism of Lipid and Glucose in Young SD Rats

XU Yun-peng, HAO Min, CHANG Xiao-tong

2015, 35(2):  97-101.  DOI: 10.3969/j.issn.1674-5817.2015.02.003

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Objective To study the effect of childhood obesity induced by high fat diet on the metabolism of glucose and lipid. Methods Thirty male SD rats (21-day-old) were randomly divided into 2 groups, and were given basic diet or high fat diet, respectively. At the 60 days old, the insulin tolerance test of rats of rats was proceeded, the body length and weight the rats were measured accurately, and the blood was collected for the measurement of biochemical markers. After the rats were sacrificed, the livers of rats were collected, sliced and stained for pathological observation. Result The obesity rat model induced by high fat diet was established successfully. The serum level of triglyceride (TG) and and total cholesterol (TC) in obese rats were higher than those of non-obese rats (P<0.05), but there were no differences in alanine aminotransferase (ALT), fasting blood glucose (FBG), high-density lipoprotein cholesterol (HDL-C), and insulin (Ins) between groups (P>0.05). At each time point, there was no difference on the insulin tolerance test. Under the microscope, there were numbers of fat bubbles in liver cells of young obese rats. Conclusion Compared with the non-obese young rats, the metabolism of lipids (TG, TC) of the obese young rats was disordered, the level of FBG and ALT showed an increasing trend, the level of HDL-C and Ins showed a decreasing trend. Obesity induced by high fat diet can lead to fatty liver of young rats.



Changes of Blood Lipid, Blood Glucose and Serum Cytokines in Different Stages of Diabetic Mice

YANG Hua, LIU Fang, ZHOU Wen-jiang, ZHOU Xiao-hui, REN Xiao-nan

2015, 35(2):  102-106.  DOI: 10.3969/j.issn.1674-5817.2015.02.004

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Objective To compare the changes of blood lipid, blood glucose, serum cytokine interleukin - 6 (IL 6) and interleukin 10 (IL-10), tumor necrosis factor - (TNF-α), a gamma interferon (IFN-γ) in mice with type 2 diabetes mellitus (T2DM) during early or late stage, and discuss the meanings of cellular immune function changes in T2DM mouse model. Methods Fourty-six male C57BL/6 mice at the age of 3 weeks were randomly divided into control group and model group (DM group). Animals in the model group were intraperitoneally injected with 50 mg streptozotocin(STZ)for 3 days and fed with high-fat diet. After 6 or 24 weeks injection, glucose tolerance, blood lipid (TG, CHOL, HDLC, LDLC) level and the contents of serum IL-6, IL-10, TNF-α, IFN-γ were measured and analyzed respectively. Results Six weeks after injection of STZ, abnormal glucose tolerance was observed in DM mice, with the significant higher level of lipid and serum IL-6, TNF-α, IFN-γ, IL-10 compared with normal control group (P<0.05 or P<0.01). With the development of disease process, the glucose and lipid metabolic disorder aggravated in DM group after 24 weeks of injection, and the levels of serum IL-6, TNF-α, IFN-γ, IL-10 were significantly higher than those of 6 weeks (P<0.01). Conclusion The STZ induced mouse model showed similar disease process in terms of blood glucose, blood lipids and cytokines changes in the different courses, which might be used as the potential disease animal model.



Evaluation on Effects of Chase and Distance-play Tests on Predicting Training Success Rate for Guide Dogs

ZHANG Ya-li, ZHOU Zi-juan, YU Jian-xiong, HAN Fang, WANG Liang, JIANG Xin, WANG Ai-guo, WANG Fu-jin, WANG Jing-yu

2015, 35(2):  107-113.  DOI: 10.3969/j.issn.1674-5817.2015.02.005

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Objective To establish the methods and evaluation criterion for Chase and Distance-play tests of dog mentality assessment (DMA) and detect their effects on predicting the training success rate for guide dogs. Method The methods of Chase and Distance-play test and the related evaluation criterion of 9 behavioral variables in total were improved and established. The behavioral variables were Following I, Following II, Grabbing I, Grabbing II in Chase test and Interest, Aggression, Exploration, Tug-of-war, Play Invitation in Distance-play test. Twelve guide dogs and 12 eliminated dogs from Chinese Guide Dogs Training Centre of Dalian were used to examine the predicting effects of the 9 behavioral variables on the training success rate for guide dogs. Result According to the results of statistical analysis, the “Play Invitation” in Distance-play test showeda significant difference between guide dogs and eliminated dogs (P=0.045). However, no significant difference was detected for the scores of the rest variables (P>0.05). To analyze the distribution of the numbers of guide dogs and eliminated dogs in different score grades, we divided the scores of each variable into 3 grades (1~2 points, 3~4 points and 5 points). For Play Invitation, the eliminated dogs were mainly concentrated in 1~2 points. For Aggression, the guide dogs and eliminated dogs were mainly concentrated in the 1~2 points. For Following I, more guide dogs distributed in 5 points grade compared to eliminated dogs. For Grabbing II, eliminated dogs were mainly concentrated in 3~4 points. For Exploration, the guide dogs were mainly concentrated in the 3~4 points. In comparison, the eliminated dogs were mainly concentrated in 1~2 points. Conclusion We have established the methods and evaluation criterion for Chase and Distance-play test of DMA. The “Play Invitation” is confirmed as an effective variable index to predict the training success rate for guide dogs.



Application of Vascular-access-port in Intravenous Glucose Tolerance Test in Minipig

CHEN Liang, PAN Yong-ming, CAI Yue-qin, ZHU Ke-yan, XU Xiao-ping, CHEN Min-li

2015, 35(2):  114-119.  DOI: 10.3969/j.issn.1674-5817.2015.02.006

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Objective To improve the reliability of intravenous glucose tolerance test (IVGTT) in minipigs and reduce the stress response during the test with vascular-access-port (VAP), and provides a new method which was fast and convenient and stable for IVGTT in minipigs. Methods The 12 Bama minipigs were randomly divided into the implantable VAP group and the normal control group, the difference of the two groups animals in IVGTT were compared, and the stress index were evaluated. Results VAP group and the control group could appear glucose tolerance test reactions in IVGTT, changes and the trend in serum insulin levels were not significantly different, but the glucose levels in VAP group at 0 min, 60 min, 90 min and 120 min were significantly lower than those of control group (P< 0.05, P<0.01), and significantly higher than that of the control group in 5 min (P<0.01); During the test, the CK levels in the control group were significantly higher than the VAP group at each time point (P< 0.05, P<0.01); ACTH and CORT levels in 0 min ,15-120 min in control group were higher than VAP group, and in 60-120 min had a significant difference (P<0.05, P<0.01) Conclusion Compare with conventional methods for IVGTT, VAP implantation way to do IVGTT suffered less stress in minipigs, more in line with animal welfare, and the test results were more reliable and stable.



Investigation on Helicobacter spp. Infection in Rat, Mouse, Guinea Pig and Rabbit

FENG Jie, ZHANG Rong, XIE Jian-yun, LI Rui-jiao, HU Jian-hua, GAO Cheng

2015, 35(2):  120-124.  DOI: 10.3969/j.issn.1674-5817.2015.02.007

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Objective To survey the Helicobacter spp. among rats, mice, guinea pigs and rabbits in Shanghai and surrounding area. Method The ileocecal region contents of rats, mice, guinea pigs and feces of rabbits were collected, and the bacterial genomic DNA were extracted. Both traditional PCR and multiplex PCR test were performed to investigate the Helicobacter spp. infection. Result The positive rate of Helicobacter spp. in rat, mouse, guinea pig and rabbit were 50.5%(105/208), 37.9%(204/538), 36.7%(36/98) and 4.9%(12/247) respectively. In addition, the triplex PCR test results of H.hepaticus, H.bilis, H.rodentium showed that H.hepaticus, H.bilis, infection can be detected in rabbit. H.bilis, H.rodentium can be detected in guinea pig. H.hepaticus, H.bilis, H.rodentium can be detected in rat and mouse. Conclusions There are different degrees of Helicobacter spp. infection in rats, mice, guinea pigs and rabbits of Shanghai and surrounding area.



Application of Concept Map in Laboratory Animal Science Teaching

WEI Sheng, LI Zi-fa, SONG Chun-hong, XU Kai-yong

2015, 35(2):  129-132.  DOI: 10.3969/j.issn.1674-5817.2015.02.009

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The concept map is used as a tool to organize and characterize knowledge and the strategy of concept map is a new way of teaching reformation. Therefore, the authors give a precise introduction of the connotation of concept map, the build process of laboratory animal science course concept map, and describe the concept map strategy for laboratory animal science teaching in order to resolve the beginners accepting difficultly, teachers teaching struggling and other key issues always harassing laboratory animal science teaching and learning.



Genetic Engineering Mice Models of Depressive-like or Depression-resistant Phenotype

WANG Xiao-jie, ZHANG Zhi-jun, YE Dong-qing

2015, 35(2):  142-148.  DOI: 10.3969/j.issn.1674-5817.2015.02.012

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The development of knockout or knockdown mice, showing a depressive or depression-resistant phenotype, have allowed us to better understand the complex relationship between genes, behavior and acquired disposition in mood disorders. Preclinical and clinical studies have established that a dysfunction of serotonin (5-HT) neurotransmission is a common hallmark in depression. Genetic mice models with a gene manipulation of?transporter, receptor, protein, enzyme gene in 5-HT system show depressive or depression-resistant phenotype. The present review revises ten genetically engineered mice models with mood alteration (either includes 5 genotypes) and summarizes their each physiological fuction,molecular mechanism, behavioristics,neurophysiology and even those translated to human. In turn, findings from human studies have helped to design and generate genetic mice models to explore new hypothesis of the etiology of human depression, and most important, to explore and refine new strategies for antidepressant medication.

Preliminary Study on Development of bcr Promotor Driven EGFP Expressing Transgenic Mice

LIU Lei, WANG Bao-zhu, LAO Quan-heng, LIU Min, XUE Zheng-feng, JI Ming-chun

2015, 35(2):  149-154.  DOI: 10.3969/j.issn.1674-5817.2015.02.013

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Objective To establish the bcr promoter drived green fluorescerce protein(bcr-EGFP) transgenic mouse model and to analyze the specificity of bcr promoter for further in vivo study. Methods The 1.1 kb bcr promoter was obtained from the genome of K562 cells (chronic myeloid leukemia cell line), at the same time, two restriction sites of Ase I、EcoR I were introduced into the up and down streams of this promoter. The CMV promoter of pEGFP-N1 plasmid was replaced by 1.1 kb bcr promoter to restructure pbcr-EGFP plasmid. Transfect the new recombinant plasmid into K562 cells and NIH3T3 cells by liposomes, after 24 hours, the expression was observed with fluorescentce. The results showed that this vector was constructed successfully. The bcr-EGFP recombinant fragment was obtained from pbcr-EGFP plasmid, and used to establish bcr-EGFP transgenic mouse by microinjection and the integration of target gene was detected by PCR. Results Total of 583 eggs of C57BL/6 were microinjected with the bcr-EGFP fragment. These eggs were transferred into 30 foster female mice, and 26 of which were pregnant and 90 pups were obtained, among which 3 pups (1♀, 2♂) were transgenic mice. Conclusion The successful establishment of bcr-EGFP transgenic mouse model has paved the way for future study of the specificity of bcr promoter.



Effect of Maternal Diabetic Environment on Development of Early Mouse Embryo by in vitro Fertilization

QIAANG Su jing, TAO Ling-yun, LIU Li-jun, XU Ping, GAO Cheng

2015, 35(2):  155-160.  DOI: 10.3969/j.issn.1674-5817.2015.02.014

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Objective The experimental mice were induced to be diabetes mellitus (DM) mice through streptozotocin (STZ) intraperitoneal injection, in order to study the effect of maternal hyperglycemia on early mouse embryo development by in vitro fertilization. Methods ICR mice aged 6~8 weeks were intraperitoneally injected with STZ to established DM models, and the normal mice were considered as control group . The model mice were superovulated by intraperitoneal injection with PMSG and hCG, then the oocytes were fertilized with sperms from normal male mice in vitro. The rates of fertilization were counted and 2-cell embryos were transferred to pseudopregnant mice. Finally 14- dold embryos were obtained to extract. total RNA. After detected by Agilent 2100 Bioanalyzer system, total RNA were reversed transcription, labeled, hybridized, eluted and the data of microarray was scanned by Affymetrix Scanner 3000. Basic data were read by Command Console Software 3.1. The experiments were repeated three times, then the expression of genes between treated group and control group were compared. Results The ratio of two-cell embryo in hyperglycemia group (52.3%), was significantly lower than that in the control group (77.2%) (P<0.01). A total of 121 differentially expressed genes (P<0.05) were screened out. One hundred and nineteen genes expression levels of the experimental group was less 0.5 times than those of the control group, while 2 genes showed a more than 2-fold up regulation. Conclusion After oocytes of model mice fertilizing with sperms of normal male mice in HTF, the rate of embryo developing to two-cell was lower than normal mice. The higher of blood glucose level, the lower of two-cell rate. The growth and development of the oocytes were affected by maternal diabetes environment through up-regulating the metabolism-related genes and down-regulated development-relating genes .



Study on Relation between Cardiovascular System and Cardiac Autonomic Nervous Function in WHBE Rabbits Myocardial Ischemia Model of Phlegm-blood Stasis Syndrome Type

PAN Yong-ming, CHEN Liang, XU Jian-qin, CHEN Min-li

2015, 35(2):  161-166.  DOI: 10.3969/j.issn.1674-5817.2015.02.015

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Objective To observe the relation between cardiovascular system and cardiac autonomic nervous function in the WHBE rabbits myocardial ischemia model of phlegm-blood stasis syndrome type, by high fat diet feeding and coronary artery ligation. Methods WHBE rabbits were randomly divided into the normal control (control) group, myocardial ischemia (MI) group and myocardial ischemia of phlegm-blood stasis syndrome (PBSS-MI) group, 6 in each group. Myocardial ischemia of MI group and PBSS-MI group were induced by ligated the coronary artery, and then PBSS-MI group was fed with high fat diet, while control group and MI group were fed with common diet for 4 weeks. The electrocardiogram was monitored by non-invasive physiological telemetry system at 2 and 4 weeks, and the changes of cardiac autonomic nervous function were analyzed by HRV methods. The cardiac hemodynamic parameters were monitored at 4th week, and using stepwise multiple regression analysis to evaluate the relation between cardiovascular system and cardiac autonomic nervous function. Results Compared with control group, HR, Max_ST segment, T_ST segment, LVEDP and Tau in MI group and PBSS-MI group were significantly increased, while LVSP, LVDP, +dp/dtmax and Cdp/dtmax were significantly decreased (P<0.05, P<0.01), and DBP, SBP and MBP in PBSS-MI group were also decreased (P<0.05, P<0.01); HRV analysis showed that the time-domain analysis indexes (SDNN, SDANN, SDANN Index, RMSSD, Tr.Ind, pNN50) and frequency-domain analysis indexes (TP, ULF, VLF, LF and HF) were significantly decreased (P<0.05, P<0.01), while LF/HF was significantly increased at 2 or 4 weeks(P<0.05, P<0.01). Furthermore, LVEDP in PBSS-MI group was significantly higher than that of MI group (P<0.05), while +dp/dtmax was significantly lower than that of MI group (P<0.05), and the degree of its reduced HRV was more serious than that of MI group. The stepwise multiple regression analysis showed that it was closely relationship between HRV parameters and +dp/dtmax, -dp/dtmax, SBP, LVSP (P<0.05, P<0.01), and also showed that the low RMSSD was an independent predictor factor of cardiovascular events. Conclusion The cardiovascular autonomic nervous function of WHBE rabbit was obviously disordered after myocardial ischemia, mainly including reduced HRV and abnormal cardiac function, and it was existed obvious correlation between the left ventricular dysfunction and HRV, and phlegm-blood stasis syndrome was more serious than blood stasis, the low RMSSD can be considered an independent predictor factor of cardiovascular events, it indicated that the deterioration of cardiovascular function was related with the decrease the excitability of the vagus nerve.

Evaluation on Analgesic Effect of Tongjingting Paste in Rats Model with Experimental Dysmenorrhea

LIU Jin, SUI Zai-yun, HOU Wen-jing

2015, 35(2):  167-169.  DOI: 10.3969/j.issn.1674-5817.2015.02.016

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Objective To evaluate the analgesic effect of tongjingting paste in rats model with experimental dysmenorrhea and to reveal its analgesic mechanism. Methods The experimental dysmenorrheal model in rats was induced with oxytocin. A writhing test was employed to evaluate the analgesic effects of tongjingting paste, which was designed into different doses, in animal subjects. The contents of 6-keto-PGF1α in the blood plasma were observed by ELISA assay. Results Tongjingting paste has a significant analgesic effect on experimental dysmenorrheal model in rats. Latent period was delayed significantly, the number of writhing response was evidently decreased as well. Besides, 6-keto-PGF1¦Á were significantly increased in the treated groups. Conclusion Tongjingting paste had a significant analgesic effect on experimental dysmenorrheal model in rats. Its analgesic mechanism may be mediated by regulating the plasma levels of 6-keto-PGF1α.