Laboratory Animal Science

Establishment and Preliminary Application of ELISA for Detecting Antibody to Felid Herpesvirus 1 in Cat

2014, 31(04): 1-5.

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Abstract: Objective To establish a ELISA method for detection of Felid herpesvirus 1( FHV-1) antibody in cat． Method Raises the CRFK cell，vaccinate the FHV-1 virus，prepare the normal CRFK antigen and special FHV- 1 antigen，titrate the best working density of enzyme union and the normal or special antigen and carry out the experiments of accuracy，sensitivity，stability and specificity． Result The best working density of normal，special antigen and the enzyme union is 0. 05 μg /mL，10 μg /mL and 1∶ 5 000 respectively; The inter-assay coefficient of variation of normal antigen and special antigen is 8. 7% and 5. 8% respectively，the intra-assay average coefficient of variation is 11. 9% and 6. 6% respectively; The detection sensitivity is 1∶ 6 400． There is no cross-reactivity with Feline panleukopenia virus ( FPV) and Feline calicivirus virus ( FCV) ． The stability test shows the relative deviation is below 25% ． Conclusion The ELISA method is good in duplication，stability，specificity，and sensitivity，as a result of which ELISA can be used in detecting the Cat FHV-1 immune body．

Investigation and Analysis of Pathology on Health Status of Beijing Laboratory Rodents in Spring 2014

2014, 31(04): 6-10.

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Abstract: Objective In May 2014，our lab has taken samples from 14 units to investigate the health status of Beijing laboratory rodents． Method We collected the heart，liver，spleen，lung，kidney，large intestine and small intestine，and put these organs into 10% paraformaldehyde solution for fixation，and then prepared into two different sections for optical microscopy observation including all paraffin specimens stained with H． E． and the frozen sections stained with Oil Red O and PAS． Result The vast majority of laboratory rodents were pathologically diagnosed as healthy animals． Liver cell swelling( 32. 6% ) ，slight lung lesion( 24% ) and protein cast of kidney( 13. 2% ) were main lesions which might affect the health of laboratory rodents． Comparing with the results of the last two years，Oil Red O staining positive rate( 6. 2% ) ，which was lower than 17. 1% in May 2013， 6. 25% in October 2013 ，26. 7% in May 2012 and 14. 8% in October 2013 had a tendency to reduce At the beginning ，kidney lesion was only found occasionally in mice ( 2% ) in 2012 and rats ( 20% ) in 2011，but became common in mice( 6% ) ，rats( 12% ) and guinea pigs( 20% ) ． Kidney lesion． changes from accidental occurrence to frequent occurrence，showed that prevention should be strengthened． Conclusion The vast majority of laboratory rodents were pathologically diagnosed as healthy animals． The result might be related to breeding and feeding environment and transport stress，and the improvement of management level and avoidance of stress can ameliorate the health status of laboratory rodents．

Effect of Salvia miltiorrhiza on Learning and Memory and Acetylcholinesterase in Brain and Serum of Mice in AD Model

2014, 31(04): 11-13.

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Abstract: Objective To evaluate the effects of Salvia miltiorrhiza on the ability of learning and memory and acetyl cholinesterase ( AchE) level in brain tissue and serum of aluminium chloride( AlCl3 ) caused AD mouse model． Method The AD mouse model was prepared by intragastric administration of 400 mg /kg AlCl3 for 60 days． Model group ( To fill the stomach and AlCl3 equal dose of normal saline． ) Salvia miltiorrhiza treatment group ( lavage AlCl3 + Salvia miltiorrhiza 1 mL． ) Salvia miltiorrhiza group( lavage salvia miltiorrhiza 1 mL) ． 24 h After the Morris water maze training，learning and memory function of the mice were tested and the levels of acetyl cholinesterase were measured． Result Salvia miltiorrhiza treatment group and Salvia miltiorrhiza group compared with normal control group，the escape latency in mice and the acetyl cholinesterase levels in the brain tissue and serum no obvious change，there was no statistically significant difference ( P ＞ 0. 05) ． The escape latent period in model group，compared with salvia miltiorrhiza treatment group，was prolonged significantly ( P ＜ 0. 05) ． The acetyl cholinesterase levels of AD mouse model in brain tissue and serum were higher than other groups( P ＜ 0. 05) ． Conclusion Salvia miltiorrhiza can improve the learning and memorizing ability of AlCl3 caused AD model mice and lower the acetyl cholinesterase levels in brain tissue and serum，which are the potential mechanisms of its prevention and treatment effects of alzheimer＇s disease．

Effects of Cortex Phellodendri Chinensis Decoction on Anti-inflammatory and Anti-bacteria

2014, 31(04): 14-17.

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Abstract: Objective To observe the anti-inflammatory and antibacteria effect of Cortex Phellodendri Chinensis decoction( CPCD) ． Method 50 Kunming mice were randomly divided into 5 groups，The mice in the study groups were administered intragastrically with CPCD，the outer ears of all mice were inflamed in left as experimental group and the right ears were not treated as a control group after treated 7 days，and then，each ear of all mice were calculated the swelling and suppression mice; 40 SPF SD rats in the left and right forelimbs were made granulomas， and then all rats were randomly divided into five groups based on the weight，and the rats were administered intragastrically with CPCD within 7 days，then，the weight of each group granulomas were calculated; 60 Kunming mice were randomly divided into six groups according to sex and body weight，all groups mice were administered intragastrically with CPCD within 7 days according to the corresponding dose，and then，each group were administered with intraperitoneal injection of Staphylococcus aureus bacteria in the first seven days of administered after 1 h，in the first seven days of administered 6 h the mice were administered once more，mortality of mice was observed in 72 hrs． Result Experimental groups were compared with control groups，each dose group which was administered intragastrically with CPCD could inhibit ears swelling of mice in different levels，the difference was statistically significant ( P ＜ 0. 05 or P ＜ 0. 01) ，of which the best group was low-dose group; Experimental groups were compared with the control group，low-dose groups which was administered intragastrically with CPCD could significantly reduce the weights of rat granulomas ( P ＜ 0. 05) ，and high dose group and medium dose group could not significantly reduce the weights of rat granulomas which were compared with the control group，the differences were not statistically significant ( P ＞ 0. 05 ) ; The mice were treats with high，medium and low-dose groups，the mortality of mice were 50% ，40% ，20% ，which were compared with the control group，the low-dose group can reduce the mortality of the mice ( P ＜ 0. 05 or P ＜ 0. 01 ) ． Conclusion CPCD had anti-inflammatory and antibacteria effect．

The Decontamination of Rag2 Knockout Rats by Caesarean Section

2014, 31(04): 18-20.

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Abstract: Objective To decontaminate Rag2 knockout rats by caesarean section． Method The caesarean section was taken on the 20th and 22th day after vaginal plugs detection，the pups were genotyped by PCR after 6—8 days of birth，and pathogens were tested at 6—8 weeks． Result The survival rate was significantly increased when the operation took on the 22th days after vaginal plugs detection，the average rate of weaning is 84. 48%． Pathogens test results met the National SPF standards of experimental animal． Genotyping results were positive，and got homozygote offspring after one time of cross． Conclusion SPF homozygote offspring of Rag2 knockout rats were achieved by caesarean section successfully．

Comparison of Three Acute Lung Injury ( ALI) Rat Model Induced by Different Etiological Factors

2014, 31(04): 21-23.

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Abstract: Objective To compare three acute lung injury ( ALI) rat models induced by different drugs，including hydrochloric acid ( HCl) ，oleinic acid ( OA) and lipopolysaccharide ( LPS) ． Method 24 female SpragueDawley rats were randomly divided into 4 groups: control( NS) ，HCl，OA，LPS． ALI model in HCl group was induced by hydrochloric acid via tracheal instillation，in OA group and LPS group were respectively induced by oleinic acid and lipopolysaccharide via vena caudalis，and in NS group was injected normal saline as the control． Carotid Blood samples were obtained for blood gas analysis after modeling 4 hours． Pathological changes of lung tissue were observed with HE staining and then scored by pathological injury． Result In comparing with control group，PaO2 was significantly lower in three ALI rat models ( P ＜ 0. 01) ，which was lower in OA group than that in HCl group ( P ＜ 0. 01) and lower in HCl Group than that in LPS group( P ＜ 0. 01) ． pathological injury score was highest in OA Group，and lowest in LPS group． Conclusion The development of ALI was closely related to the etiological factors． ALI induced with different causes has different features． The score of lung injury was OA group ＞ HCl group ＞ LPS group．

Establishment of Rat Model of Chronic Fatigue Syndrome

2014, 31(04): 24-27.

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Abstract: Objective To establish the rat model of chronic fatigue syndrome( CFS) by using congestion and sleep disturbances． Method Rats were crowded in a small box，were interfered normal sleep，modeling for 1 month． The mental state，skin and hair，body weight，food intake，water intake were obserred，and conducted exhaustive swimming experiments． Result After modeling，rat mental malaise，aggressive and confrontational behavior disappears; loose skint，withered body hair，thinning off; body weight，food intake，exhaustive swimming time were reduced． The symptom of highly congested group was the most typical，general performance score reaches 5. 6． Body weight，food intake，swimming time reduced most obvious． Conclusion We succeeded in establishing a new animal model of CFS in rat by using congestion and sleep disturbances．

Cryopreservation of Mongolian gerbil 2-cell Embryos by Vitrification based on EG with Two Steps

2014, 31(04): 28-31.

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Abstract: Objective Using the thyleneglycol( EG) -based solutions with two steps to freeze the 2-cell embryos of Mongolian gerbil( Meriones unguiculatus) and to appraise the effect of embryo cryopreservation． Method The embryos were exposed serially to EFS20 for 3 minutes and EFS40 for 1 minute． The embryo trealed were immersed into liquid nitrogen． Result The embryo recovery rate was 86. 73% and after freezing-thawing it survival rate was 82. 94% ． And about 30% of the freezing-thawing embryos developed to blastocyst in vitro． Conclusion This results demonstrates that Mongolian gerbil embryos can be safely cryopreserved using vitrification solutions based on EG with two steps．

The Influence of Osphyalgia Strapping on Prolapse Lumbar Intervertebral Disc Model in Rabbit

2014, 31(04): 32-35.

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Abstract: Objective To observe the influence of osphyalgia strapping on rabbit prolapse lumbar intervertebral disc model． Method With the surgical method，the round filter paper was placed on axillary region of sacral nerve root． In order to observe the influence of osphyalgia strapping on rabbit prolapse lumbar intervertebral disc model.Result Paralysis recovery，pain recovery and touch recovery of medication administration groups were all ahead of time in comparison with model group，and the recovery ratio was higher． The recovery of high dose group was the best with the positive rate reaching more than 90% from complete paralysis to semiparalysis and from semiparalysis to recovery; in middle and low dose group，the filter paper weight decreased obviously，whole blood viscosity reduced sharply，and erythrocyte electrophoretic time shortened evidently． Conclusion Osphyalgia strapping had the role of lightening nerve-trunk edema，hyperemia，denaturation，foreign body grannuloma formation; promoting nerve fiber’s recovery and nerve-trunk’s conduction，improving lower limbs’paralysis; and osphyalgia strapping had the role of improving microcirculation and hemorheology with increasing the blood flow．

Construction of Resource Database for Animal Models of Human Disease

2014, 31(04): 36-40.

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Abstract: Construction of resource database for animal models of human disease is to enable people to quickly and accurately screening resources in animal models，classification，to facilitate the researchers chose an animal model needs and can provide quick contact animal model units，saving query time，improve the efficiency of scientific research． The database is divided into spontaneous animal model database，induced animal model of the database and the database genetically engineered animal models of three parts，each part contains an animal model name， number，animal information，preparation of model information，provide the model unit of information，support literature information and so on． By functions and query the database to build，in addition to provide users with the search function，to obtain useful information，the animal models of human disease also resource development and resource sharing of information has far-reaching significance．

Comparison of Sterilization Effect of Silver Nitrate with Conventional Antibiotics on Bacteria and Fungi

2014, 31(04): 41-45.

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Abstract: Objective To investigate whether the silver nitrate has the advantages over conventional antibiotics in sterilization of common bacteria and fungi． Method Plate dilution method was performed，that is，three drugs were dissolved in the culture medium at different concentrations，and then the bacteria and fungi were plated． The minimum inhibitory concentration was determined by growth situation of the bacteria and fungi． Result Silver nitrate had similar inhibitory function to both bacteria and fungi ( P ＞ 0. 05 ) and the minimum inhibitory concentration was between 3 － 7 μg /mL． For antibiotics，the minimum inhibitory concentration of neomycin to S． aureus and K． pneumoniae was between 0.1 － 1.0 μg /mL and that of gentamicin to S．aureus and E．coli were 0.1 μg /mL and 1.25 μg /mL，respectively． However，there was no inhibitory function was found for gentamicin against P．aeruginosa，and the inhibitory function of neomycin against P．aeruginosa was 16 μg /mL． Conclusion There is no selectivity of inhibitory effect for silver nitrate against bacteria and fungi． Comparatively，the antibiotics prefer to inhibit the bacteria，but not fungi． With the bactericidal and bacteriostatic effects of the silver particles and silver ions are re-realized in the modern medicine，the combination of silver nitrate and antibiotics may be an effective therapy strategy in clinical．

The Operating Experience for TUNEL Staining of Apoptotic Cell

2014, 31(04): 46-48.

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Abstract: TUNEL Staining of apoptotic cells have been widely used in scientific research． Due to the multi-step of TUNEL staining，the fine of operation，the multi-factors of DNA fragmentation，the experiments were failed． In this paper，they were expounded on the details of tissue’s collection，tissue’s size，fixative’s selection，tissue’s ischemia time，tissue in fixative’s time and the experimental operation．

The Establishment and Operation of Conservation Facilities for SPF Rats and Mice

2014, 31(04): 49-53.

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Abstract: The conservation and breeding of rats and mice is a prerequisite for expand production and application of rats and mice． The genetic traits，animal quality and the production output of rats and mice were directly affected by the quality control of the conservation and breeding． Our center established the conservation facilities in 2012 year，and conserved 7 rats and mice strains． So far，conservation animals maintain high quality，and are in line with SPF animal standard． This paper introduces the establishment of conservation facilities for SPF rats and mice， animal strains，quality requirements，facility operation and assessment，and operation experiences in our center，in order to provide the reference for related work of rats and mice conservation and breeding．

Application of CRISPR-Cas System in Biology Research

2014, 31(04): 54-58.

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Abstract: Clustered regularly interspaced short palindromic repeats ( CRISPR) system was a new genetic modification tool． Its advantages were short work time，low cost，high efficiency and so on． Here we review the CRISPR-Cas system structure，development history，classification，mechanism and application in biology research．

The Progress in Rodent Models of Hyperuricemia

2014, 31(04): 59-62.

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Abstract: To establish an ideal animal model of hyperuricemia is needed to explore the pathogenesis machanism and find a drug to control and treat the hyperuricemia． Scientist have made certain progress for the study of animal model of hyperuricemia． And in these models，rodents were chose greatly． To manufacture the rodent model of hyperuricemia，the theory of increasing the source of the uric acid，reducing uric acid excretion and inhibiting uricase were used． This review will summarize the choice of rodents，induced drugs and the methods．

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