Laboratory Animal Science

Screening with Some Blood Biochemistry Parameters in GUIZHOU Minipigs and GUANGXI Bama Minipigs

2017, 34(04): 1-3.

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Abstract: Objective To screening the minipigs on hand to find genetic diseases for further selected breeding． Method Thirteen of blood biochemistry parameters were detected in 60 GUANGXI Bama minipigs and 50 GUIZHOU minipigs． The parameters which exceed the mean ± 2SD range were regarded as abnormal ones． And the abnormal parameters were detected and analyzed repeatedly．Ｒesult Forty-two abnormal minipigs were found with 19 GUIZHOU minipigs and 23 GUANGXI Bama minipigs among them． After repeating the detecting and analysis， 2 Bama minipigs ( No． 37 and No． 103) were chosen as seeds for further selected breeding for atherosclerosis resisted and susceptible strains． Conclusion After screening with blood biochemistry parameters and repeated detection and analysis，2 Bama minipigs were selected as presumed atherosclerosis resisted and susceptible breeding seeds．

Neurotoxic PrP106-126 Fragmentinduces Neuropathological Changes in Primary Cortical Neurons

2017, 34(04): 4-10.

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Abstract: To investigate the neuropathological damage of prion diseases，we observed the apoptosis-associated protein changes by immunoblottingand the morphological changes of primary cortical neurons ( PCCN) exposed to PrP106-126 by transmission electron microscopy ( TEM) ． Furthermore，PrP106-126-stimulated primary neuronal death was verified by TUNEL and Annexin V-FITC /PI apoptosis assay． After exposure to PrP106-126 for 24 h，the PCCN showed ultrastructural pathology similar to typical “spongiform vacuoles”in TSEs． A large number of vacuoles of various sizes and swollen and disorganized smooth endoplasmic reticulum ( SEＲ) appeared in the cytoplasm． Nuclear fragments and condensed chromatin were released into the cytoplasm． Consistent with previous researches，we confirmed that PrP106-126 peptide induced mitochondrial cytochrome c release and caspase 3 activation，increased Bcl-2-associated X protein ( BAX) and reduced B-cell lymphoma protein 2 ( Bcl-2) in a timedependent manner． Our observations proved that PrP106-126 is a valid model for researching PrPSc neuropathological damage and neurotoxicity，which provide the theoretical basis for the researchin prion diseases and associated neurodegenerative diseases in vivo.

Comparison of the Different Model Establishment Methods of Kidney Yang Deficiency in Mice Caused by Hydro-cortisone

2017, 34(04): 11.

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Abstract: To compare the advantages and disadvantages between different dosage and methods of administration of cortisone in deficiency of Kidney Yang animal models，we used intragastric administration，intraperitoneal injection and subcutaneous injection ways to mice with different doses ( 12. 5，25，50 mg / kg) of hydro-cortisone for 9 days to investigate the variations on mortality rate，general state，weight，anus temperature，autonomic activities，swimming duration and viscera index． After 9 days，general status in mice is poorer，the 25，50 mg / kg dose group are more obvious． Besides subcutaneous injection of 25 mg / kg group，other groups weight was significantly lower than before building; besides intragastric and intraperitoneal injection of 12. 5 mg / kg，subcutaneous injections of 25 mg / kg group，number of autonomic activities in mice decreased to some extent; in addition to the various ways of 12. 5 mg / kg group，the swimming time was significantly shortened than the blank group; intraperitoneal injection and subcutaneous injection of 25 mg / kg group spleen index，all ways of 12. 5 mg / kg group of adrenal gland index compared with the blank group has no obvious change，the rest of the group of spleen，thymus and adrenal gland index were significantly reduced，seminal vesicle and preputial glands of each dose group have no obvious change． So we synthesized the inspection indexes，intragastric administration 25 mg / kg dose group is superior to that of other groups． All the indexes can show the signs and symptoms of Kidney Yang Deficiency，it is more reliable Kidney Yang Deficiency model method．

Tocotrienol Plus Oxaliplatin Improve Immunity Function in Mice with Colitis-associated Cancer

2017, 34(04): 15-20.

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Abstract: Objective To observe the effect of tocotrienol plus Oxaliplatin on mice with colits-associated cancer． Method Male ICＲ mice ( 5weeks old) were randomly divided into 5 groups( n = 20 /group and 10 mice per cage) ，respectively control group，AOM /DSS group，AOM /DSS + OXA group ( 3 mg / kg·bw OXA) ，AOM /DSS + T3 group ( 100 mg / kg·bw T3) and AOM /DSS + OXA + T3 group ( 3 mg / kg·bw OXA and 100 mg / kg·bw T3) ，all animals were sacrificed at the end of the study ( week 20) ． During the course of the experiment，mice were monitored by continuous body weight measurement; At sacrifice，recorded the mice weight and the weight of spleen，colon，calculated the index of spleen and colon，observed the pathological changes of spleen and colon in mice，evaluated the production of inflammation cytokines and chemokines ( TNF，IL-6，IL-4，IL-2) in blood of mice．Ｒesult Compared with control group，the index of spleen and colon was significantly increased in AOM / DSS group，P ＜ 0. 05; Compared with AOM /DSS group，AOM /DSS + OXA and AOM /DSS + OXA + T3 could markedly decline the index of spleen，and the index of colon was decreased in AOM /DSS + OXA + T3 group，P ＜ 0. 05． AOM /DSS + OXA，AOM /DSS + T3 and AOM /DSS + OXA + T3 could inhibit the expression of TNF，and the expression of IL-6 was remarkably decreased in AOM /DSS + OXA，AOM /DSS + OXA + T3 groups; Compared with Control group，AOM /DSS could decline the production of IL-4． Conclusion Tocotrienol and Oxaliplatin were able to improve immunity function，inhibited the development of colorectal cancer．

The Study on Blood Collection Method in Mouse in China Journal Ｒeports near 5 Years

2017, 34(04): 21-24.

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Abstract: Objective To investigate the domestic situation of method of blood collection in mouse． Method The data was retrieved from Wan Fang database in the search term“mouse“and“blood”as defined by the 2012 to 2016 Journal Citation Ｒeports． Perform a statistical analysis on the articles which meet the criterion．Ｒesult Four hundred and forty-eight articles were retrieved，and 444 articles met the criterion． The most used three blood collection method are orbital sinus ( 33. 3% ) ，picking eyeball ( 22. 6% ) and tail vein ( 15. 4% ) ． Heart blood with anesthesia was 60 percent，and orbital sinus blood with anesthesia was 3. 2 percent． Conclusion The utilization rate of picking eyeball method is high． Given that it can cause great pain in mice，we suggest that replace this method with jaw venous plexus in pursuance or other method ． The rate of heart blood and orbital sinus with anaesthesia is not high． So it should be taken into account．

Analysis for Detection Ｒesults of The Air Borne Bacteria in Quiescent State and Dynamic Statein Negative Pressure Barrier Facility

2017, 34(04): 25-31.

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Abstract: Objective To understand the situation of the air borne bacteria in quiescent state and dynamic state in negative pressure barrier facility and analyze parameters changing． Method The air borne bacteria in quiescent state and dynamic state were tested according to the national standards GB 14925—2010 for laboratory animal environment and facility ．Ｒesult The air borne bacteria were 100% above standard for breeding area in barrier environment; The air borne bacteria were 92. 86% for the other area of pollution corridor and the air borne bacteria were 91. 67% for the second dressing room in barrier environment． Conclusion Although there was no national standard of the air borne bacteria in dynamic state in barrier environment，It is necessary for detecting the air borne bacteria，The accumulation of testing data of four years which will provide reliable evidences for the standardization of barrier environment．

Study on Anti-fatigue and Anti-oxidant Effects of Shenzujing Tablets in Mice

2017, 34(04): 32-35.

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Abstract: Objective To explore anti-fatigue and anti-oxidant effects of Shenzujing tablets in mice． Method One hundred and twenty mice were randomly divided into 5 groups: control groups，Liuwei Dihuang pill group and shenzujing tablets ( 0. 5，0. 25，0. 125 g / kg) groups． Each study group contained twenty-four mice． The mice received daily oral reatments of drugs for 7 days． The control group received daily oral treatments of isovolumic distilled water for 7 days． The 2 h after the last treatment，50% of mice tail root were randomly selected to load 2 grams of lead wire，the exhaustion swimming time were observed． The other 50% of the mice tail root were loaded 0. 4 grams of lead wire，after 1 hour swimming，SOD and MDA content in myocardium and muscle were detected． Another 144 mice were randomly divided into 6 groups: control group，model group，Liuwei Dihuang pill group and Shenzujing tablets ( 0. 5，0. 25，0. 125 g / kg) groups; The administration method were similar as above． After the last treatment，50% of mice were randomly selected and given intragastric administration of 0. 4 mL / 50% ethanol， serumand liver MDA，SOD content were determined，brain tissue of the other 50% mice were made into 10% brain homogenate and put Fe2 + － VitC solution in，hatched for 20 mins at temperature of 37 ℃，brain MDA，SOD content were determined．Ｒesult The mouse swimming exhaustion time in Shenzujing tablets ( 0. 25，0. 125 g / kg) groups were significantly prolonged( P ＜ 0. 05) ，and the myocardial SOD activity were significantly improved( P ＜ 0. 05) ． The MDA contents in biceps femoris muscle were lowered ( P ＜ 0. 05) ． serum and liver MDA concentration in alcoholic liver injury mice were lowered ( P ＜ 0. 01) ，SOD concentration were increased ( P ＜ 0. 01) ; MDA content in mice of the brain injury induced by Fe2 +－ VitC were reduced( P ＜ 0. 01) ． Conclusion The shenzujing tablets can increase SOD activity and decrease MDA accumulation and have the ability to maintain the body's metabolism，increase the mice swimming exhaustion time，good anti-fatigue effect，its effect is closely related to the anti oxidant effect．

Animal Experimental Study on Characteristics of Intervertebral Disc Healing in Different Periods after Annulus Fibrosus Suture

2017, 34(04): 36-42.

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Abstract: Objective Preform the animal experimental study to research the characteristics of intervertebral disc biomechanical strength in different periods after the annulus fibrosus suture． Method A total of 40 sheep were underwent retroperitoneal 3 levels exposure of the lumbar spine． The Control Group did not penetrate into the annulus fibrosus． The Ｒesearch Group penetrated into the annulus fibrosus and perform annulus fibrosus suture． In Ｒesearch Group，at 3 months after surgery，Group A were taken MＲI images and underwent biomechanical testing to assess the strength of the disc． Histology was also conducted． Group B were performed 6 months after surgery． Group C were performed 12 months after surgery．Ｒesult The biomechanical strength of the disc at 3 months after operation was( 1. 39 ± 0. 68) MPa，( 2. 04 ± 0. 88) MPa at 6 months． ( 3. 58 ± 1. 10) MPa at 12 months，and it was close to the Control Group with( 3. 90 ± 1. 59) MPa． In Ｒesearch Group，loss of intervertebral height was( 1. 01 ± 0. 29) mm after 3 months． ( 1. 90 ± 0. 65) mm after 6 months，( 2. 04 ± 0. 85) mm after 12 months． Conclusion Annulus fibrosus suture play an important role in the maintenance of the mechanical strength of the intervertebral disc，also promote the repair of annulus fibrosus． And the biomechanical strength of the intervertebral disc was correlated with the repair time of annulus fibrosus．

Cloning and Phylogenetic Analysis of Cytochrome b Gene in Tamarisk Jird( Meriones tamariscinus)

2017, 34(04): 43-49.

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Abstract: Objective Complete sequence of mitochondrial Cytochrome b of Tamarisk jird was cloned by the PCＲ method ． The sequence was determined and used to phylogenetic analysis． Method The primer was designed according to the published sequences． The PCＲ product was sequenced and determined． The Cyt b sequences of 24 rodents were used to analyze the nucleotide composition and genetic distance． These sequences also used to construct the phylogenetic tree base on maximum likelihood and maximum parsimony．Ｒesult The Cyt b gene was cloned in Tamarisk jird，which have high homology with Mongolian gerbil and Midday jird． The result of phylogenetic tree showed that Tamarisk jird is higher genetic relationship with Meriones and Cricetidae． The result of nucleotide composition showed that nucleotide composition of Tamarisk jird showed the characteristics of mammal mtDNA; the nucleotide composition of Tamarisk jird is similar with Meriones and Ｒattus． Conclusion In this research，the genetic relationship between the Tamarisk jird with other rodents was analyzed． This research is an important work for study on evolution，structure and function of the Tamarisk jird mitochondrion．

Prevalence and Molecular Identification of Myocoptes musculinus in Laboratory Animals in China

2017, 34(04): 50-54.

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Abstract: Objective The aim of this study is to establish a method of rapid identification of Myocoptes musculinus infection，and analyze the test result of 843 batches specific pathogen free ( SPF) laboratory animal from 50 different manufacturers in China，and to provide data and technical support for the strategy of Myocoptes musculinus purification． Method Direct microscopy real-time dynamic video recording techniques combined with morphological identification method were applied to screen Myocoptes musculinus infestation． Multiple PCＲ method were used to identify the isolated Myocoptes musculinus cytb ( cytochrome b) ，cox2 ( cytochrome c oxidase subunit 2) and ATP6 ( ATP synthase F0 subunit 6) genes，and the Myocoptes musculinus infection was further confirmed at the molecular level．Ｒesult Based on the direct microscopy real-time dynamic microscopic video recording technology，39 Myocoptes musculinus positive samples were detected，the positive rate was 0. 76% ，and the morphology was the egg，larva，nymph and adult． The parasite species were preliminarily determined according to the size and morphology of Myocoptes musculinus eggs，larvae，nymphs，and adults． The cytb，cox2 and ATP6 genes of Myocoptes musculinus were amplified from the single egg，larva，nymph and adult of Myocoptes musculinus separated from the positive samples，and the result showed that 39 positive samples of morphology，molecular identification are all positive，and no cross reaction with other different species of parasites． Sequence comparison result showed that the nucleotide similarity of cytb，cox2 and ATP6 gene sequence of Myocoptes musculinus in different individual animal was 100% ． Conclusion Direct microscopy real-time dynamic microscopic video recording technology combined with multiple PCＲ technique has high sensitivity and specificity and can be used to detect Myocoptes musculinus． The test result showed that Myocoptes musculinus was positive in laboratory animal samples，suggesting that laboratory animal pathogen examination in our country should be further strengthened． This research was the first molecular identification and infection investigation of Myocoptes musculinus in laboratory animals in China．

Ｒesearch on the Standard of Microbiological Quality Control in Experimental Cattle

2017, 34(04): 55-58.

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Abstract: Experimental cattle are widely applied in different fields of research，however，the corresponding standard of microbiological quality control for experimental cattle has not been established yet． In the present study， literature analysis and Modified Delphi expert consultation method were performed as to explore the requirements and grade system establishment for experimental cattle microbiological quality control，and surveillance of local standard，further providing a reference for experimental cattle preparation and usage in Beijing．

Construction of Literature Classification Ｒetrieval Platform for Animal Models of Tumor Disease

2017, 34(04): 59-62.

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Abstract: Animal models are the important support tools of tumor research． In this study，Literature retrieval platform was developed to integrate literature resources in animal models of tumor disease，resolve the problems about disordered dispersion of their data resources，provide dynamically the latest advances on animal model research，and create a convenient，real-time，efficient，professional network information platform for researchers． Our research will lay the foundation for the further study about literature data mining on animal models， increasingly，data-analysis service in making animal models will be offered and more related characteristic data bases incrementally developed．

Investigation on the Current Situation of Laboratory Animal Production in China Mainland

2017, 34(04): 63-70.

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Abstract: The purpose of this paper was to investigate and analyze the production status of laboratory animals in China Mainland． This investigation on the mainland of China covered 31 provinces，autonomous regions and municipalities directly under the central government． The survey found that the production of laboratory animals under 1 produce license can meet the needs of experimental animal use under 3 to 4 usage licenses． The survey also found that the production of experimental animals shows the characteristic of regional distribution，which was closely related to the geographical environment and economic development． The mice，rats and chicken eggs were mainly SPF animals，the hamster was mainly clean animals，while the other animals were mainly conventional animals，so there is a large room to improve for the grade of animal production． Only 13 species of experimental animals were produced under the production license，so it was hard to meet the needs of scientists and technicians． Therefore， the development of experimental animal industry in China needs systematic plans at the national level，so as to comprehensively improve the output，species and quality of experimental animals，and eventually provide strong support for the development of science and technology in China．

Innovation and Development Mode of Laboratory Animal Center for Universities

2017, 34(04): 71-73.

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Abstract: In this paper，by analyzing the current status and existing problems of development of the laboratory animal center of universities in our country，to conduct a preliminary exploration for higher education innovation and development from the aspects of capital investment，system innovation，service content，laboratory management system，cost analysis and management，talent management mode etc．

Prospects for Immunophenotypic Analysis of Tree Shrew

2017, 34(04): 74-78.

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Abstract: Immunophenotypic analysis is extensively utilized in the clinical diagnosis of various diseases and exploration of animal models． Tree shrew，a newly established experimental animal，attracts more attentions in the biomedical field． However，in the tree shrew models，we have difficulties in exploring the further mechanisms of diseases，particularly performing immunophenotypic analysis，due to the limitation of commercial reagents． Antibody plays critical roles in developing the platform of immunophenotypic analysis． Emerging evidences from studies of antibody cross-reactivity in other experimental animals indicate that by screening the commercial antibodies，it is possible to find antibodies that could react with antigens from other species． This provides a novel strategy to develop the platform for immunophenotypic analysis of tree shrew，which will be beneficial to further application of tree shrew as experimental animals．

The Development of Animal Model for Food Allergy

2017, 34(04): 79-82.

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Abstract: Food allergy is a common disease with high incidence． As the essential method for the study of food allergy，animal models are increasingly important． Although there has been some food allergy models for clinical diagnosis and research，they are not integrated． Herein，we review the design of animal models for food allergy． The dose threshold could obtain through the dose-sensititzation relationships to evaluate the allergic reaction． As the processing of protein changed，the sensitizability of the protein will changed as well． Here，the design of various animal models are reviewed， including among others considerations of species and strain， diet， route of administration，dose and formulation of the test protein，relevant controls and endpoints measured． Through the analysis and summary of the above aspects，this paper was expected to provide reference for the design of animal model for food allergy．

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