Laboratory Animal Science

Microsatellite Analysis in 3 Kunming Mice Outbred Colonies

SHANG Hai-tao~1, WEI Hong~1, YUE Bing-fei~2, XU Ping~3

2009, 26(02): 1-6.

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Objective Providing valuable information and references for standardization of Kunming mice by detecting and analyzing genetic profiles, genetic variation and genetic differentiation of Kunming mice populations. Methods 15 microsatellite markers were screened by a fluorescence-based semi-automated genotyping method for the 3 populations of Kunming mice, BJ(population of Kunming mice maintained at the Rodent Laboratory Animal Resources in Beijing),SH(population of Kunming mice maintained at the Rodent Laboratory Animal Resources, Shanghai branch) and F1(Hybrid population between SH and population from the First Biochemical-pharmaceutical Factory of Shanghai), and genetic variation of each population and genetic diversities between populations were evaluated. Results A total of 92 alleles were detected in the 3 populations, with 2～13 and mean 6.13 at each locus, and the mean unbiased expected heterozygosity was 0.5721, which implies that there is abundant genetic variation in the populations of Kunming mice. 61, 63 and 51 alleles were detected, and mean unbiased expected heterozygosity were 0.4923, 0.5177 and 0.4550 in population BJ, SH and F1, respectively. It suggested that slightly more polymorphisms were detected in population SH than in population BJ and less polymorphism were detected in population F1. The most, 43 shared alleles, were detected between population SH and F1, which suggested low genetic differentiation between them. 37 alleles were detected between population BJ and SH, and 32 alleles were detected between population BJ and F1, which suggested high genetic differentiation. Poor genetic differentiation was discovered between SH and F1 with the mean Fst being 0.0222 and Nei's unbiased measures of genetic distance estimates being 0.0279; moderate genetic differentiation was discovered between BJ and SH with the mean Fst being 0.1433 and Nei's unbiased measures of genetic distance estimates being 0.3881; large genetic differentiation was discovered between BJ and F1 with the mean Fst being 0.1667 and Nei's unbiased measures of genetic distance estimates being 0.4162. Population SH was clustered with population F1 firstly, which were clustered with population BJ then by UPGMA method. Conclusion Genetic profiles of 3 Kunming mice were identified by 15 microsatellite loci, and moderate or large genetic differences between populations from different keepers were validated, and loss of alleles should be avoided during hybridization between populations for standardization of Kunming mice. The present study could provide valuable information and references for establish standard population of Kunming mice and genetic monitoring for it.

Breeding,Reproducing,and Identifying for PAX-8 Gene Knock-out Mice

CHEN Tong-ke~1, CHEN Xi-wen~1, GUAN Min-qiang~1, ZHAO Hui-ling~1, WANG Li-hua~2, LIN Xiao-kun~3, LI An-le~1

2009, 26(02): 7-9.

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Objective To breed and identify PAX-8 gene knock-out mice. Methods Heterozygote mice were bred and reproduced.Wild type, heterozygote genotype,and homozygote genotype would appear in the offspring of parents. Genome DNA extracted from the mice's tails was subjected to PCR test for genotype identification. Male heterozygote mice were selected to mate with the female heterozygote mice to acquire homozygote baby mice according to Mendel law. Results The breeding and reproducing were successful and more heterozygote genotype mice were reproduced. Conclusions Appropriate methods of breeding, reproducing, and identifying are effective ways to gain PAX-8 gene knock-out mice from heterozygote mice.

Influences of Taking Long-term ZhenTian Capsule on Blood Biochemistry and Blood Cell Composition in Rats

WANG Ting~1, SONG Huai-yan~1, ZHAO De-ming~2

2009, 26(02): 10-16.

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Objective To explore the influences of ZhenTian capsule on blood cell composition, liver function, and kidney function. Methods The rats were orally given ZhenTian capsule for 180 days with the doses of 1.315,2.63 and 5.26g(crude medicinal materials)/kg(13,26.1 and 52.2 times of clinic does) while the control group was given the distilled water of the same volume and for 4 weeks of stopping administration. Meanwhile, rats' behavior and appearance were observed, and night shit and urine everyday amounts were measured. The rat's indexes of hematology and biochemistry were determined. Results All the detected indexes were in normal rang. The value of blood routine and biochemical indices in the tested group had no difference compared with controlled group. Conclusion Taking long-time ZhenTian capsule has no a bad influence on blood composition,liver and kidney functions.

Abnormal State of Coagulation in Experimental Colitis of Rats

PENG Tao,LIU Yu-lan

2009, 26(02): 17-22.

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Objective:To study the relationship between inflammatory activities and coagulation abnormal in the TNBS induced rat's colitis model. Method 30 SD-rats separated into 3 groups randomly,Include 1 normal control group: In this group every rats were given an enema with normal saline, and were killed 14 day after enema. 2 1week colitis group, in this group every rat was given an enema with TNBS, and was killed 7 day after enema. Colitis 2 weeks group: In this group every rats were given an enema with TNBS, and were killed 14 day after enema. We choose PT、APTT and the activity of AT as the index of coagulation. And we assess the level of damage and inflammatory state of the colitis rats by macroscopically score, histological score and the level of TNF-α in each group. Result Compared with normal control group, TNBS induced 1 week colitis group, and 2weeks colitis group has a shorter PT and APTT and a lower activity level of AT. The number of plate per litre is higher than the normal control group. The difference has statistical significance. And we find that index of tissue damage and inflammatory has negative correlation with PT 、APTT and AT. Conclusion There is abnormality of coagulation in the TNBS induced rat colitis,which is correspondence with the state of the illness closely. The test's result implied that abnormal state of coagulation in IBD patients have close correlation with the activity of inflammatory. And TNBS induced rat colitis model can act as a model of inflammation activate coagulation for further study in this field.

Effects of Therapeutic Dose of Pantoprazole Sodium on Rat Animal Model with Acute Reflux Esophagitis

XIANG Shao-jie, QIAO Min, MENG Li, ZHANG Hui-ying, CHEN He

2009, 26(02): 23-24.

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Objective To observe curative effect of acute reflux esophagitis with the therapeutic dose of Pantoprazole Sodium. Method To study the effect of therapeutic dose of Pantoprazole Sodium on the degree of esophageal mucosa injury with the acute Reflux Esophagitis rat animal model induced by ligation of pylorica and cardiomyotomy. Result The therapeutic dose of Pantoprazole Sodium can obviously relieve esophageal mucosa injury of rat animal model. Conclusion The therapeutic dose of Pantoprazole Sodium has curative effect on acute reflux esophagitis rat animal model.

The Impact of Hypotensor on Rat's Cardiovascular Function Induced by Nicotine

WANG Kai, LIU Ding-kuo, PEI Zhi-hua

2009, 26(02): 25-27.

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The experiment proved that elevation of blood pressure and cardio acceleration from nicotine can be blocked by the two drugs on the rat. Elevation of blood pressure from nicotine can be blocked by large dose of chlorisondamine (10 mg/kg) . Mecamylamine can only block cardio acceleration from nicotine on the rat, but long-term uses of mecamylamine do not block the effects of nicotine to cardiovascular system. The block function of effects from nicotine to cardiovascular system of chlorisondamine better than that of mecamylamine. The block function of large dose of chlorisondamine to ganglions can change the heart rate and blood pressure, and the heart rate and blood pressure changes controlled by block level of different dose of chlorisondamine and mecamylamine.

Establishment of PCR Assay for Detection of B Virus

LI Xiao-bo, XIAO Jing, FU Rui, HE Zheng-ming

2009, 26(02): 28-32.

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Objective To establish PCR assay for detection of B virus DNA. Method We designed primers, amplified B virus by PCR and verified its sensitivity and specificity.Results There are predicted products when P1,P2 or P3,P4 as primer and BV DNA for template, but there are no PCR products when other viruses DNA as templates; There are 382 bp products when P5,P6 as primers ,BV and HSVI DNA as templates. Cut by SacⅡ there are 176 bp and 206 bp products when BV DNA as template. The products cannot be cut off when HSVI DNA as template. Conclusion We discriminate B virus by PCR and distinguish B virus from HSVI.

Examining The Depressor Substance in MZR by Tiger Skin Cat

LIU Ji-feng~1, GU Yu~2, BAI Yu~1, TANG Shan-bo~2, ZHANG Li-rong~2, LIU Fu-ying~3

2009, 26(02): 33-35.

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Objective To observe the effective of Tiger skin cat on examination the depressor substance in MZR. Methods The Tiger skin cat were anesthetized and given histamine phosphate at the dose of 0.05 μg·kg~ -1 、 0.10 μg·kg~ -1 and 0.15 μg·kg~ -1 respectively by intravenous injection. Then the response value of cats' blood pressure at each dose was determined and sensitivity of cats to histamine phosphate was assessed. Histamine phosphate and MZR were injected acocording to pharmacopeia and the effects of different doses of MZR (0.05 μg·kg~ -1 、0.10 μg·kg~ -1 、0.15 μg·kg~ -1 ) on cats' blood pressure were observed. Results The value of cats' blood pressure exceeded 2.67 kPa after different doses of histamine phosphate injection and the sensitivity of cats to histamine phosphate was qualification. The changes of cats' blood pressure in different MZR were not greater then a half of that induced by histamine phosphate. Conclusions The MZR depressor substance was consistent with pharmacopeia and the Tiger skin cats could be used for the examination of the depressor substance in MZR.

The Tans-saphenous Femoral Artery Punctures Method on The China Agricultural University Mini-pigs

FENG Xue-quan, XIAO Fu-shun, LI Mu, CHEN Guang

2009, 26(02): 54-56.

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Objective To evaluate the trans-saphenous femoral artery puncture method for mini-pigs. Methods 16 China agricultural university mini-pigs, after binding and anesthesia, exposing the saphenous artery by incising local skin, 4F radial artery catheter was placed in it after puncture, then the vertebral artery catheter was placed in the femoral artery via the saphenous artery guided by the super slide angled Cook guide wire. Result The puncture was carried out 25 times in 16 China agricultural university mini-pigs, all succeeded with the success ratio 100% and the mean time required was about 15 minutes. Conclusion The tans-saphenous femoral artery puncture method with the advantages of high success rate, mini-invasiveness, convenient operation, saving time and repeat for more times is competent for mini-pigs' endovascular manipulation.

Simply Discussion on ELISA Operative Technique of Avian Disease

ZHAI Xin-yan, LU Sheng-ming

2009, 26(02): 57-59.

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Objective Enzyme-linked Immunosorbent Assay (ELISA) is a serologically detective technique having been widely adapted as the disease monitoring means. ELISA antibody test reagents for avian pathogens have been largely used in routine quality control of SPF chicken. The ELISA procedures not only has its technical request, but also is influenced by lots of factors. The wrong results will be obtained without paying attention to operation. Therefore, the key points of the ELISA operation have been analyzed and summarized while detecting avian diseases, in order to communicate and study with everyone.

The Effect of Combining Diazepam and Sodium Pentobarbital on Anesthesia in Mice

GUO De-yu, TIAN Xin, ZHU Zi-xin, YE Cui-fei, ZHENG Wen-rong, WANG Yu-feng

2009, 26(02): 60-61.

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Objective To study the effect of combinational anesthesia, the mice were injected with diazepam and pentobarbital sodium. Methods ICR mice were randomly divided into control group and the test group. Control group of mice were subcutaneously injected with normal saline (0.1 mL/10 g), and experimental group of mice were subcutaneously injected with diazepam (1 mg/kg). Spontaneous activity was examined by an apparatus to record the movement of mice in a box. And then, the two groups of mice were injected with pentobarbital sodium (50 mg/kg). The disappearing time and recovery time of righting reflex were recorded. Results The spontaneous activity in the mice injected with diazepam was significantly lower than the mice injected with saline (P<0.01). For the mice injected both diazepam and sodium pentobarbital, the time to fall asleep was significantly shorter (P<0.01) and the sleep time was significantly longer (P<0.01) than the mice injected sodium pentobarbital only. Conclusion Combination of diazepam and sodium pentobarbital can significantly improved the anesthesia effect in mice.

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