Laboratory Animal Science

Establishment and Application of Rapid Nucleic Acid Detection Methods for Mouse Hepatitis Virus

XIONG Wei, JIANG Jing, ZHANG Qiang, LIU Jun-Ping, WEI Xiao-Feng, HUANG Zhong-Rong-1, LI Jian, HU Jian-Hua

2013, 30(04): 1-5.

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Abstract: Objective To establish the methods of nucleic acid detection for mouse hepatitis virus ( MHV) so as to meet the need of rapid quarantine for rodent animals in the port of entry． Method Using primer and TaqMan fluorescent probe software，the primers and probe were designed to target conserved gene of MHV． And methods of RT-PCR and Real-time RT-PCR using TaqMan probe were established to detect MHV from the clinical samples of rodent animals． Result The two established methods for MHV detection were proved to be specific with high sensitivity and efficiency． Using these two methods to detect clinical samples collected from MHV positive mouse， the results show RT-PCR and Real-time RT-PCR methods have high stability and reliablity． Conclusion The methods of RT-PCR and real-time RT-PCR to detect MHV were established，which are suitable to detect easily available samples such as feces and urine．

Establishment and Application of Rapid Nucleic Acid Detection Methods for Mouse Hepatitis Virus

XIONG Wei, JIANG Jing, ZHANG Qiang, LIU Jun-Ping, WEI Xiao-Feng, HUANG Zhong-Rong, LI Jian, HU Jian-Hua

2013, 30(04): 1-5.

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Abstract: Objective To establish the methods of nucleic acid detection for mouse hepatitis virus ( MHV) so as to meet the need of rapid quarantine for rodent animals in the port of entry． Method Using primer and TaqMan fluorescent probe software，the primers and probe were designed to target conserved gene of MHV． And methods of RT-PCR and Real-time RT-PCR using TaqMan probe were established to detect MHV from the clinical samples of rodent animals． Result The two established methods for MHV detection were proved to be specific with high sensitivity and efficiency． Using these two methods to detect clinical samples collected from MHV positive mouse， the results show RT-PCR and Real-time RT-PCR methods have high stability and reliablity． Conclusion The methods of RT-PCR and real-time RT-PCR to detect MHV were established，which are suitable to detect easily available samples such as feces and urine．

A Survey of Growing Development and Reproductive Performance of p53 Knockout Mice

XU Meng, CHAI Meng-Yin, SHENG Bo, CHEN De-Xi

2013, 30(04): 6-8.

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Abstract: Objective To investigate the differences in growing development and reproductive performance between p53 knockout mice and BALB /c mice． Method 34 p53 knockout mice with half males and half females and 34 BALB /c mice ( half male and female) were adopted in the trial，and their body weight and body length were measured from 2 to 12 weeks of age． Litter sizes at the second parity from 30 pairs of mice were checked． Result The difference in body weight between 3 － week old p53 knockout mice and BALB /c mice was not significant ( P ＞ 0. 05) ，and body weight of 6 － week old p53 knockout mice was significantly lower than that of 6 － week old BALB /c mice ( P ＜ 0. 05) ． Body length of 3 － week and 6 － week old p53 knockout mice was significantly lower than that of 3 － week and 6 － week old BALB /c mice ( P ＜ 0. 05) ． Reproductive performance of p53 knockout mice was significantly lower than that of BALB /c mice ( P ＜ 0. 05) ． Conclusion p53 gene knockout could exert significant effect on the growing development and reproductive performance of the mice．

Establishing of Pathological Evaluation Method for the Therapeutic Effect of Dexamethasone in a LPS-induced Mouse Acute Pneumonia Model

YANG Dong, CHOU Wei-Yi, ZHANG Chang, ZENG Da-Di, WANG Shuang, SUN Zhi-Wei, ZHAO De-Ming

2013, 30(04): 9-12.

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Abstract: Dexamethasone was used as a positive drug to establish and validate a pathological evaluation method for a lipopolysaccharide ( LPS) induced mouse acute pneumonia model，and develop a low-priced and easy preliminary pharmacodynamic evaluation model for the inflammatory and infectious diseases． The LPS-induced mouse acute pneumonia model was developed by the nasal inhalation of LPS． The pathology classification standard was set up via HE staining based on the lung injury severity against time points． Dexamethasone was adopted as a positive drug to validate the pathological evaluation method． After inhalation of LPS，the lungs were collected at different time points for pathological evaluation． A 0 – 5 level pathological classification standard of lung injury was established． This method was used to evaluate the therapeutic effect of the positive drug dexmethasone，and proved a significant difference between the LPS control and dexmethasone-treatment group． We established the pathological evaluation method for the therapeutic effect of dexamethasone in the LPS-induced mouse acute pneumonia model． The method can be used for the preliminary screening of anti-inflammatory drugs and their pharmacodynamic studies．

Establishment and Evaluation of a Wedged-block-hemisected Injury Model in Rats Spinal Cord

ZHAO Fu-Sheng, WU Geng, WU Yang, WEI Shu-Fang, HAO Xin, LI Yue-Zhen, ZHANG Ji-Fei

2013, 30(04): 13-17.

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Abstract: Objective Establishment a novel hemisected spinal cord injury model with rats to supply model for clinical research． Method 25 SD rats were randomly divided into wedged-block hemisected group ( A group，n = 10) ，sham operation group( B group，n = 10) and control group ( C group，n = 5) ． After operation，the stability of the model was comprehensive evaluated by grading the scores of Basso-Beattie-Bresnahan ( BBB) ， swash plate test， HE staining and MRI assay． Result After operation，the mean scores of BBB in injured hind limb and maximum angle of swash plate in group A were significantly lower than group C and B( P ＜ 0. 05) ，but these parameters without different between B group and C group; 28 days after operation，HE staining and MRI assay shows a significant morphological changes in spinal cord tissue of group A; Conclusion The results suggested that the rat model established by this method with high quality of stability and reliability which suitable to evaluate the spinal cord injury research．

An Observation on Ethological and Pathological Changes in Rat of Parkinson＇s Disease Induced by Lactacystin

CHEN Chen, QUAN Jun, LI Man-Rong, SONG Wen, BU Ping, HU Rong

2013, 30(04): 18-21.

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Abstract: Objective To observe the ethological and pathological changes of the rats of parkinson’s disease induced by injection of lactacystin into the substantia nigra． Method Lactacystin /DMSO ( 8 μg /2 μL) was stereotacticly injected into the left substantia nigra of model group，while the DMSO was injected into the left substantia nigra into DMSO group． Apomorphineinduced rotation test was performed on day 6， 13 and 20 after the injection． Immunohistochemical staining was conducted to test the aggregation of α － synuclein and the number of TH( tyrosine hydroxylase) positive neurons． Result The rats of model group revolved rightward and the frequency exceed 7 r /min． Aggregation of α － synuclein was revealed in the paraffin section of substantia nigra of model rats and the number of TH-positive neurons were significantly reduced． Conclusion Rats model of PD can be established by stereotactic injection of lactacystin into the substantia nigra．

Discussion on Some Problems Concerning Surgical Implantation of DSI Implanter into Aortaventralis of the Rats

LUO Ding, FAN Xiao-Nong

2013, 30(04): 22-25.

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Abstract: It is now a tendency to apply the system of telemetering blood pressure for long term in the animal experiments on hypertension． The high success of implantation of DSI implanter is the key to obtaining of the animal blood pressure and performing of further experiments． The paper presented the some problems that caused the implantation of DSI implanter to fail in our experiments and the solutions to the problems so as to give concrete and specific suggestions to the researchers in this area．

An Observation on Ethological and Pathological Changes in Rat of Parkinson＇s Disease Induced by Lactacystin

ZHU Shao-Ping, HU Li, BAO Bo

2013, 30(04): 26-30.

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Abstract: Objective To observe the ethological and pathological changes of the rats of parkinson’s disease induced by injection of lactacystin into the substantia nigra． Method Lactacystin /DMSO ( 8 μg /2 μL) was stereotacticly injected into the left substantia nigra of model group，while the DMSO was injected into the left substantia nigra into DMSO group． Apomorphineinduced rotation test was performed on day 6， 13 and 20 after the injection． Immunohistochemical staining was conducted to test the aggregation of α － synuclein and the number of TH( tyrosine hydroxylase) positive neurons． Result The rats of model group revolved rightward and the frequency exceed 7 r /min． Aggregation of α － synuclein was revealed in the paraffin section of substantia nigra of model rats and the number of TH-positive neurons were significantly reduced． Conclusion Rats model of PD can be established by stereotactic injection of lactacystin into the substantia nigra． Key words: Parkinson’s disease; lactacystin; animal model

Effects of the Palygorskite Corncob Bedding on Liver Microsomal Cytochrome P450 Enzymes in Rats

SHU Chang, ZHANG Yan-Ying, WU Jian-Jun, TAO Wei-Tian, LIN Xing-Yao, ZHU Jian-Kun, KANG Wan-Rong, LI Cun-Xiang

2013, 30(04): 31-34.

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Abstract: Objective To observe the effects of palygorskite corncob bedding on the contents of cytochrome P450 ( CytP450) and cytochrome b5 ( Cytb5) ，and activity of CYP1A2 and CYP2E1 in the rat liver cells． Method 120 SD rats were randomly divided into ordinary wood shavings group，corn stalk group，palygorskite corncob group and control group． The contents of CytP450 and Cytb5，and activity of CYP1A2 and CYP2E1 were determined after 30 days，60 days，and 90 days of feeding． Result After 60 days and 90 days of feeding，the difference in CytP450 content between ordinary wood shavings group and control group and between corn stalk group and control group was significant ( P ＜ 0. 05) ，however，the difference was not significant between palygorskite corncob group and the control group ( P ＞ 0. 05) ． The differences in liver microsomal Cytb5 content，and CYP1A2 and CYP2E1 activity were not significant among the groups ( P ＞ 0. 05) ． Conclusion The palygorsklte corncob did not exert inhibiting effect and inducing effect on liver microsomal CytP450，Cytb5，CYP1A2 and CYP2E1，and ordinary wood shavings had stronger inducing effect on rat liver CytP450 and corn stalk had weak inducing effect on rat liver CytP450．

Effects of Chronic Fluorosis on Rat Growth and the Antagonism of Mg2 + ，Al3 + and Ca2

YIN Li-Jing, ZHAO Run, ZHOU Jie, HE Yuan-Qing-1

2013, 30(04): 35-38.

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Abstract: Objective To study the effects of chronic fluorosis on the growth of rate and the antagonism of Mg2 + ， Al3 + and Ca2 + to fluorine toxicity so as to provide the data for the researches on the molecular mechanism and the treatment of endemic fluorosis． Method 24 the first week postweaning male SD rats were randomly divided into 4 groups ( n = 6 in each group) ，namely the control group ( group 1) ，fluoridating group ( group 2) ，fluorine + Mg2 + + Al3 + group ( group 3) and fluorine + Ca2 + group ( group 4) ． The water added with fluorine，with fluorine + Mg2 + + Al3 + and with fluorine + Ca2 + was respectively given to the rats in the different groups for three months． The body weight，food intake and the amount of water drinking were measured and recorded every two days，and the 24 － hour urine was collected daily to determine the fluorine content in the urine during 3 days before sacrifice． Result The speed of weight gain in group 4 was significantly higher than that in group 1，and the speed of weight gain in group 2 and group 3 was significantly lower than that in group 1． Urine fluorine content of group 2 was higher than that of the other 3 groups，while that of group 3 and group 4 was lower than group 2 but higher than group 1． There was no significant difference in food intake and amount of water drinking among groups． Conclusion The chronic fluorosis could decelerate the growth rate，while adding Mg2 + ，Al3 + or Ca2 + into the drinking water，especially Ca2 + ，may antagonize this effect．

The Organ Changes and Occurrence of Spontaneous Tumor in Long-term Breeding Wistar Rats and Their Death Causes

XIAO Cheng-Rong, TAN Hong-Ling, MA Zeng-Chun, WANG Yu-Guang, LIANG Qian-De, TANG Xiang-Lin, ZHAO Yong-Hong, GAO Yue

2013, 30(04): 39-43.

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Abstract: Objective To observe the weight of viscera and organ coefficient，and occurrence of spontaneous tumor of survival and dead Wistar rats during 2 years experiment，and investigate their death cause． Method The gross anatomy and histopathological examination for the rats sacrificed at the end of the two-year experiment and the dying or dead rats during the experiment were carried out instantly． Their body weight and weight of their brain，heart， lung，liver，spleen，kidney，adrenal glands，thymus，ovary，uterus，testis，and epididymis were measured and organ coefficients were calculated，and occurrence of spontaneous tumor was recorded． Result There were significant differences in weight of body，brain，heart，lung，liver，spleen，kidney，adrenal glands and organ coefficients of brain and adrenal glands between female and male rats survived for 104 weeks． The gross anatomy and histopathological examination revealed that 80. 5% of rats died of dyscrasia and multiple organ failure ( MOF) resulted from tumor lesions，17. 1% of rats died of MOF resulted from non-tumor lesions，and 2. 4% of rats succumbed to MOF caused by other lesions． There was no significant difference in death cause between male and female rats． Conclusion The present study provided background information for carcinogenicity tests performed on Wistar rats，and the tumor lesions are the leading death cause of Wistar rats．

Exploration on Three Common Methods of Multiple-drawblood in Rat

WANG Ming-Li, ZHANG Jing, WANG Yao-Hui, LI Shu-Lian, MA Yuan-Fang

2013, 30(04): 44-45.

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Abstract: Objective To study the commonly used three kinds of method for rat venous blood sampling a day or in a short period of time． Method Total 90 male Wistar rats were used． Three methods were used for the blood sampling，angular vein blood sampling method，the jugular vein blood sampling method and the cardiac puncture blood sampling method． Result 0. 2—0. 3 mL blood can be collected from the angular vein everytime，and 3— 4 times a day; 0. 2—0. 4 mL blood can be collected from the jugular vein everytime，and 6—8 times a day; 0. 2— 0. 4 mL blood can be collected from the cardiac everytime，and 3—5 times a day． Conclusion Large volume and high quanlity of blood samples can be collected by the three blood sampling methods，which can be used in the rat experiment．

The Essentials and Experience of Developing Model of Cerebral Ischemia-reperfusion by Suture-occluded Method in Rat

HE Fang-Yan, HAN Chun-Ni, LI Yan, LIN Qing

2013, 30(04): 46-48.

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Abstract: The method of developing model of cerebral ischemia-reperfusion in the rats by suture-occlude method had the advantages of no need for craniotomy，controllable duration of ischemia，and more definitive infarct location． At present the method has been extensively used in China and abroad for studying ischemia-reperfusion injury． A comparative study on developing model of cerebral ischemia-reperfusion in the rats by suture-occlude method has been carried out in our laboratory． This paper presented the essentials and experience of developing model of cerebral ischemia-reperfusion by suture-occluded method in the rat．

Parasites of Dog in China and Laboratory Animal-standards and Monitoring for Parasitology

WANG Ming-Shuang, ZHANG Xiao-Hong, TIAN Zhi-Ning, WANG Yan-Ping

2013, 30(04): 49-53.

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Abstract: List of parasites which parasited in dog has been known in China is given and route of infection and prevent point are discussed in this paper． Some suggestions about standards for parasitology of dog and monkey on China’s Nation Standard GB 14922 － 1—2001 are proposed ，which can be used to discuss for other scholars．

Application of SPF Chicks in Prevention and Control of Avian Diseases

DI Xin-Yan, QU Ping, MA Ying, ZHANG Qian

2013, 30(04): 59-61.

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Abstract: SPF chicks means specific pathogens free chicks． The source of SPF chicks is definitely clear． SPF chicks are not able to be detected the primary avian pathogens． SPF chicks are produced by seed breeding，raising in clearing facilities and microorganism monitoring． With the development in the past 50 years，SPF chicks play a vital role in not only life science research ，but also avian disease prevention and control，especially used in the avian disease research，vaccine production and assessment，diagnostic reagents preparation，as well as therapeutic goods production，et al．

Introduction to Four Rodent Laboratory Animal Model Strains

KANG Feng, ZHANG Hai-Feng, WANG Long

2013, 30(04): 62-64.

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Abstract: A lot of laboratory animal models have been developed to prevent and treat cardiovascular diseases such as hypertension． Four rat model strains about hypertension including Spontaneous Hypertensive Rats ( SHR) 、 Wistar Kyoto Rats ( WKY) 、Dahl /Salt Sensitive Rats ( DSS) and Salt Sensitive-Chr 13BN Rats ( SS-13BN ) will be introduced in this paper．

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