Laboratory Animal Science

Characteristics of Blood Biochemistry and Leukocytes in EBV Transgenic Mice

KANG Ai-jun1, GUO Chang-zhan2, TIAN Feng3, WANG Xue-wen3, ZHENG Zhen-hui3, ZHAO De-ming1

2010, 27(01): 1-4.

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Objective To examine some serum biochemical indexes and the characteristic of leukocytes in EBV transgenic mice. Methods GPT, GOT, TP, ALB, BUN, CRE, TC, TG and GLU of sera collected from two-month and four-month-old EBV transgenic mice as well as normal KM were measured. The number and classification of the whole-blood WBCs were also determined. Results There were significant differences in TP, ALB, BUN, CRE, TC, and TG levels between EBV transgenic and KM mice. The obvious differences were also observed in the number and classification of WBCs between EBV transgenic and KM mice. Conclusion The serum biochemical indexes and characteristic of leukocytes determined by present study provide important basic references for scientific research and application of EBV transgenic mice.

Sperm-mediated Gene Transfer By Injection of Lentiviral Vector Into Mouse Siminiferous Tubules

MA Xiao1, YE Hua-hu2, DU Xiao-yan1, WANG Ying1, CHEN Zhen-wen 1

2010, 27(01): 5-9.

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Objective To investigate the practicability of lentiviral vector-based sperm-mediated gene transfer in production of transgenic animals.Methods According to the titer of lentiviral vector (LV) administrated, 4～5 week-old mice were divided into two groups: high-titer (2 mice) and low-titer (6 mice) groups, into which lentiviral vectors carrying green fluorescent protein (LV-GFP) were injected through the seminiferous tubules at the concentration of 20μL/testis, and the administrated titer of LV was 1×109 and 2×10 7 TU/mL for the two groups respectively. Sampling was conducted at the 4th and 8th week (high-titer group), and the 5th, 13th and 17th week (low-titer group) after injection. Transgenic mice were examined by PCR, and the expression of GFP was detected by fluorescence microscopy and immunohistochemistry.Results (1) positive rate of GFP gene in mice testis was 62.5%; (2) weak expression of GFP was only observed in high-titer group, and mainly distributed in the based-membrane of seminiferous tubules and the interspaces between tubules; (3) injection into seminiferous tubules caused some damage, but did not affect the reproductive capacity of mice. Conclusion Lentiviral vectors can successfully transfect mice testis in SMGT and the expression of GFP gene depends on the titer of LV administrated.

Pathological Observations of Main Arteries in Twelve-month-old ApoE～(-/-) Mice

TIAN Feng1, KANG Aijun2, ZHANG Kuo2, ZHOU Shu-pei2, ZHENH Zhen-hui2, ZHAO De-ming 1

2010, 27(01): 10-12.

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Objective To investigate the pathological characteristic of main arteries in twelve-month-old ApoE-/-mice. Methods Both male and female ApoE-/-mice of twelve-month-old were selected in this study. Frozen sections were made from the aortic root, carotid artery, abdominal artery and renal artery, and stained with oil red O for examining the formation of atherosclerosis spots. Results Lipid plagues were detected in aortic roots and renal arteries of all ApoE-/-mice, and atherosclerotic plaques were observed in abdominal arteries of four ApoE-/-mice. No plaques were detected in carotid arteries of all mice. Conclusion ApoE-/-mice are the excellent animal model for atherosclerosis research.

Study on the Parameters of Main Internal Organs in SPF Male SD Rats at Different Ages

WANG Yao 1,2 , TANG Da-xuan1, GE Lin, XIONG Jing-yue1, WANG Li 1,2 , LIANG Ya-jun 1,2 , MOU Dao-hua 1,2 , TAN Zheng-huai 1

2010, 27(01): 13-15.

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Objective To investigate the normal values of body weight, indexes and weight of main internal organs in male Sprague Dawley (SD) rats at different ages.Methods Weight of body and ten internal organs of male SD rats were measured monthly from 2 to 8 month-old, and the indexes of each organ were calculated simultaneously. Results From two-month to eight-month old, the weight of heart, liver, kidney, brain, epididymis and adrenal gland increased, but decreased in thymus gland. Meanwhile the indexes of heart, liver, spleen, lung, brain, thymus gland and adrenal gland reduced gradually as the body weight increased.Conclusion In SPF male SD rats, the weight and indexes of the main internal organs can reflect their structure changes and physiological functions.

Effect of Different Types of Fatty Acids on Serum Biochemical Indicator and Insulin Resistance in SD Rats

GAO Qing-song, LIU Yuan, ZHANG Tao, ZHAO Shuang, WANG Zhi-hong

2010, 27(01): 16-21.

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Objective To analyze the effect of serum biochemical indicator and insulin resistance in SD rats induced by four types fatty acids: medium-chain saturated fatty acids (MC-SFA, MCF group), long-chain saturated fatty acids (LC-SFA, LCF group), n-6 polyunsaturated fatty acids (n-6 PUFA, SUF group), and n-3 polyunsaturated fatty acids (n-3 PUFA, TUF group). Methods Normal male SD rats were fed on standard laboratory chow (8.23% of calories as fat) or isocaloric high fat diets (44.75% of calories as fat) for 10 weeks. Serum glucose (GLU), insulin, triglyceride (TG), cholesterol (CHO) and HDL-cholesterol (HDL-c) were measured with an interval of 18 days. The insulin sensitivity was estimated by ISI index (ISI= ln 1/( FPG×FINS)). Results After 10 weeks, body weights of LCF and SUF group were obviously higher than those of normal and other high-fat groups. Serum TG was obviously higher in LCF group compared with other high-fat groups P<0.05). High-fat feeding groups, especially the TUF group, exhibited obviously lower HDL-c (P<0.05) than normal group; Serum insulin significantly increased in LCF group compared to other groups (P<0.05); Both LCF and TUF group showed lower ISI than normal group (P<0.05); However there was no significant difference in GLU among the five groups. Conclusion Body weight, serum biochemical indicator and insulin resistance of rats are affected significantly by the types of fatty acids.

A Chronic Gastritis Rat Model Induced by Combined Treatment of Chemicals

CHEN He, ZHANG Hui-ying,LIU He, QIN Wen-yan,QIAO Ju-jiu, QI Yue, XIANG Shao-jie, Bao Yu-long, CUI Ming-min, SONG Da-fu

2010, 27(01): 22-24.

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Objective To establish a chronic gastritis rat model.Methods Healthy Wistar rats (male∶ female=1∶1) weighting 180～220 g were randomly divided into control and model group. The model group was administrated intragastrically with ethanol and sodium deoxycholate to induce chronic gastritis.Results After 3 months, chronic gastritis symptom was successfully produced in the model group.Conclusion Intragastric administration of sodium deoxycholate combined with ethanol is suitable for making chronic gastritis model in rats. This model is reliable and repeatable for traditional Chinese medical study in chronic gastritis.

Effect of Guben Qingfei Tiegao on Guinea Pig Model of Allergic Asthma

LI Xian-hua, SONG Da-fu, GAN Yu, DU Jia-lin, QIAO Shi-ju ZHOU Shu-mei, GU Xiao-ying

2010, 27(01): 25-26.

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Objective By using the guinea pig model of allergic asthma, to determine the effects of Guben Qingfei Tiegao on relieving asthma and anti-inflammatory.Methods The guinea pigs were allergized by intraperitoneal injection with 10% ovalbumin normal saline solution. Fourteen days later, they were stimulated with 1% ovalbumin normal saline 3 times. After treating with the Guben Qingfei Tiegao, the latency period of asthma and the amount of WBC in bronchovesicular irrigating solution were detected.Results Guben Qingfei Tiegao could prolong the latency period of asthma and reduce the amount of WBC in bronchoalveolar irrigating solution in asthmatic guinea pigs.Conclusion Guben Qingfei Tiegao has positive effects on anti-asthmatic and anti-inflammatory in guinea pig model of allergic asthma.

Analysis of Microsatellite DNA Polymorphisms and Genetic Monitoring of Four Inbred Lines of Zebrafish

MU Yuan 1,2 , SUN De-ming2

2010, 27(01): 27-30.

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Objective To analyse microsatellite DNA polymorphisms in genetic monitoring of four inbred lines of Zebrafish.Methods 3 microsatellites DNA loci on different chromosomes in four inbred lines of Zebrafish(AB,TU,WIK,LF)were investigated by PCR analysis.Results Three microsatellites DNA could be amplified efficiently, and displayed polymorphism among the various lines.Conclusion Analysis of microsatellites DNA polymorphisms by three microsatellites DNA is a fast and economical way for genetic monitoring of AB,TU,WIK,LF inbred Zebrafish.

Effect of Econazole Nitrate Vaginal Tablet on Rabbit Model of Candida Albicans Vaginitis

DU Jia-lin, Qi Yue, LI Xian-hua, XIANG Shao-jie, ZHANG Hong, JIA Dong, YUAN yuan

2010, 27(01): 31-33.

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Objective To observe the efficacy of econazole nitrate vaginal tablet to cure the inflammation in rabbit model of candida albicans vaginitis.Methods The rabbit model of candida albicans vaginitis was produced by infection of the domestic rabbit vagina with the white beads germ. The reducing rate of the white beads germ in the vagina secretion and the recovering degree of the vagina mucosa organization were examined.Results and Conclusion Econazole nitrate vaginal tablet can inhibit growth of the white beads germ, and remove many pathological changes, such as vaginal hyperemia,dropsy,erosion and increased secretion, at the dose of 23.5 mg/Kg、47 mg/Kg、94 mg/Kg. There is no significant difference in efficiency between econazole nitrate vaginal tablet and clotrimazole, but different significantly between econazole nitrate vaginal tablet and excipient.

Detection of Serum Antibody in Beagle Dogs Vaccinated with Canine Adenovirus Type 1

KANG Ji-ping, LI Qiu-bo, FENG Xiao-ke, YANG Jie

2010, 27(01): 34-36.

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Objective To establish a proper procedure for vaccination regimen against infectious canine hepatitis (ICH) in Beagle dogs.Methods Canine hepatitis hemagglutination inhibition (HI) antibody of 120 serum samples collected from 24 Beagle dogs (20-day to 14-week old) was examined by the indirect hemagglutination inhibition test.Results Maternal HI antibody titer was 1∶8-1∶32 (average 1∶20) and 1∶2-1∶8 (average 1∶5) in 20-day old and 35-day old puppy respectively. HI antibody titer was 1∶8-1∶32 (average 1∶14) 3 weeks after the first immunization, and 1∶32-1∶128 (average 1∶45) 3 weeks after the second immunization and 1∶64-1∶256 (average 1∶79) 3 weeks after the third immunization, respectively.Conclusion The Best Combinational Canine Attenuated-live Virus Vaccines are preferred to prevent canine hepatitis, and the first immunization should be performed in 35-40-day old. The second and the third immunization should be performed at an interval of 3 weeks after the first and the second immunization respectively.

Monitoring and Evaluation of Environmental Factors of Minipig Facilities in Beijing

WANG Yan-rong, SUN Shu-hua, MENG Jin-ping,YANG Xu, LIU Yun-bo*

2010, 27(01): 37-39.

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Objective To monitor the environmental factors of experimental minipig facilities in Beijing and provide the basic data for local environmental and housing standard setting for experimental minipigs. Methods Examinations were conducted according to the appendix A-H of National Standard in Experiment Animal Environment and Facilities (GB14925-2001). The environmental factors, such as temperature, humidity, work-lighting, animal-lighting, noise, airflow velocity, ventilation, fallen bacteria, air cleanness, pressure, ammonia contents (dynamic), in 5 conventional and 3 barrier facilities, were examined. Results All the environmental factors were controlled at the range set by the National Standard (GB14925). Conclusion The environmental factors controlled in minipig facilities in Beijing meet the GB14925-2001, and it is necessary to give much more attention to the environmental requirements of minipigs in different growth stages.

Management System of the Laboratory Animal Facility of Peking Union Medical College Hospital

ZHANG Dong

2010, 27(01): 40-42.

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Based on the characteristics of researches in Peking Union Medical College Hospital and animal experiments, management system of the laboratory animal facility was explored and discussed.

A Novel Automatic Litter Suction Equipment Driven by Vacuum Vortex Pump for Laboratory Animal Barrier Facilities

WANG Long, LI Hong-cai, LU Sheng-ming

2010, 27(01): 43-44.

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Wide-range disposal of waste bedding affects negatively on sanitation of barrier facilities. Therefore, it is very essential to develop an automatic litter suction equipment driven by vacuum vortex pump for reducing spread of the micro-organisms in barrier facilities, keeping healthy of laboratory animals, saving labor and consumables costs, as well as maintaining environmental sanitation inside and outside the barrier facilities.

Animal Models of Glaucoma

WANG Xiao-lei,ZHANG Xiu-lan

2010, 27(01): 45-52.

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Glaucoma is the second leading cause of blindness in the world, however, the mechanism of glaucomatous optic neuropathy remains unclear. Animal model is a very useful tool to study the etiology and therapy strategy. A variety of laboratory animals, such as rat, rabbit and primate, has been used to mimic specific aspects of human glaucoma progression. This review describes the progress on glaucoma animal models, including the newly produced transgenic mouse models, such as gene targeting glaucoma model and CLR (calcitonin receptor-like receptor) acute angle-closure glaucoma model. The advantage and limitation, and the pathophysiological changes in retinal ganglion cells, optic nerve and retina of each model are also discussed.

Review of Animal Models for Depression

QIN Qin, LIU Li-xue

2010, 27(01): 53-59.

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Commonly used animal models of depression in the pharmacological studies are reviewed, and their application and mechanism are also described. Development of the further models of depression is proposed simultaneously.

Genetic Breeding of Guangxi Bama Mini-pig

WANG Ai-de, LAN Gan-qiu, GUO Ya-fen

2010, 27(01): 60-63.

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Objective To develop a mini-pig strain with small body size, neat coat color, high reproductive performance and high genetic stability. Method 1) In the closed pur-bred breeding population (A strain), random mating method was used. 2) The animals of zero generation of inbreeding strain (B strain) was from the 10 th generations of closed pur-bred breeding group .The full-sib mating method was used to develop the inbreeding strain. 3) Directional selection was carried out in the selection programme. 4) The genetic stability was monitored by genetic marker analysis. 5) Some animal’s biological characters and nutritional requirement were determined. Results:1)The experimental mini-pig strain named as Guangxi Bama mini-pig was successfully developed in 1994 and continuous genetic breeding was carried out for 22 years. 2)The random mating had been continuously done for 21 generations in closed pur-bred breeding population until 2009. 3) The inbreeding strain had been cultivated for 11 generations until 2009. 4) Manifest selection progress was acquired on body size, neat coat color and reproductive performance. 5) The average observed number of 19 microsatellite loci in the 17 th generation of closed pure-bred population was 3.00. The average observed number of alleles,effective number of alleles and average rate of homozygous of 21 microsatellite loci in the 7 th generation of inbreeding group were 1.9048, 1.5461 and 0.7232, respectively. 6) The protein, digestible energy, calcium and available phosphorus requirement for the grower animal were determined to be 12%, 12.95Mj/kg, 0.65% and 0.32%, respectively. 7) The local standards of Guangxi Bama mini-pig strain proposed in the research was issued and published by the People's Government of Guangxi Zhuang Autonomous Region in 2008. 8) Guangxi Bama mini-pig was evaluated as excellent model animal in the study of cardiovascular diseases and diabetes by many research institutions in China. Conclusion We have successfully cultivated inbreeding and closed colony mini-pigs with unique characters, which are excellent laboratory animal used for medical and biological researches.

Determination of IgG in Rat Serum by Liquid Chromatography

YANG Xu, WANG Yan-rong, MENG Jin-ping, SUN Shu-hua, LIU Yun-bo

2010, 27(01): 64-66.

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Objective To establish a method for separation and determination of rat serum IgG using the high performance liquid chromatography (HPLC). Methods A HI-Trap Protein G column was used as the analytical column, and gradient elution was proformed using the mixture of phosphate buffer and Tris-Glycine buffer as the mobile phase according to the characteristics of the samples. The target compound was detected at the adjustable wavelength of 280nm. Results The detection limit was in the range of 0.5～1.5 mg/mL. Taking the signal-to-noise ratio (S/N) of 10 as the detection limit, the lowest examination density was 0.31 mg/mL, and the average rate of recovery in rat serum was 90.5%. Conclusion The results show that HPLC method is feasible, rapid, simple and accurate for separation and determination of serum IgG.

Comparison of Three Methods to Stain Mouse Vaginal Smears

DING Yu-long, SUN Li, LI Li-ya

2010, 27(01): 67-69.

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At present, mouse estrous cycle can be identified by examination of the cells in vaginal secretions, and different methods have been developed to stain those cells. In this study, Wright's stain was proved to be the best one among HE, Wright’s and Pap stain methods.

A Practicable Method for Perfusion and Fixation of Large Animals

ZHOU Chuan-long, FANG Jian-qiao, SHAO Xiao-mei

2010, 27(01): 70-71.

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To avoid losing the antigens, animal bodies have to be perfused and fixated after sampling. The large animals are unusual to be used as experimental objects, so the development of perfusion and fixation in large animals is insufficient. In this communication, perfusion and fixation methods for large animals are compared and discussed.

Application of Propofol-combined Anesthesia in Cardiac Catheterization of Minipigs

WANG Yang, WANG Cheng-li, ZHANG He

2010, 27(01): 72-73.

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Objective To investigate the effect of propofol-combined anesthesia in minipig cardiac catheterization. Methods 148 healthy minipigs (2-3 month-old and 15-25kg weight) were used for 350 times in the present study. Atropine and morphine were administered intramuscularly at the concentration of 0.3mg/kg and 5mg/kg respectively 30 min before the experiment. Somnolence happened 10 min after intramuscular injection of 0.2mg/kg midazolam. Tracheal intubation was performed after slow intravenous injection of 2.5-3.5mg/kg propofol. Results The propofol-combined anesthesia is a good method with shorter induction time, quicker reaction, quick and stable recovery and less side effects. Conclusion The propofol-combined anesthesia appears to be a satisfactory method for cardiac catheterization in Minipigs.

Design of A Frequency Converted Pressure Water Supply System for Monkey House Washing

WANG Hai-feng1, WU Bao-jin1, RAO Jun-hua1, ZHANG Tai-hua1, TIAN Sen-lin 2

2010, 27(01): 74-76.

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To overcome the limitations of washing methods commonly used in non-human primate breeders, a frequency converted pressure water supply system for monkey house washing was designed. This system has the characteristic of soft starting, controllable constant pressure, and sectionalized-time control. Many advantages of this system, such as high water impact pressure, high efficiency, and water saving, make it practical in monkey house for daily washing.

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