Laboratory Animal Science

2016, 33(02): 1-5.

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Abstract: Objective To observe the effect of diabetes mellitus ( DM) on the septic inflammation． Method Diabetic rat model was induced by high glucose and high fat diet and streptozotocin( STZ) injection． On this basis，the sepsis model by the cecal ligation and puncture ( CLP) ． Then the heart tissue and whole blood specimens were harvested and inflammatory factor TNF-α was assayed by ELISA，neutrophil percentage was calculated，and observe the heart tissue was stained by periodic acid-Schiff stain ( PAS) ．Ｒesult ( 1) Compared with normal rats，diabetic rats typically emerged polyuria，polydipsia，polyphagia，and weight loss，with a significant rise in blood glucose and weight loss ( P ＜ 0. 001) ． PAS stain showed myocardial cells staining in normal-control rats，without interstitial cells and blood vessels staining，but cardiomyocytes and interstitial microvessels in diabetic rats all stained． ( 2) In sepsis model based on the diabetes，after CLP the rats showed abnormal activities and mental state; in normal-CLP rats serum TNF-α values were significantly higher than those in normal-control rats ( P ＜ 0. 01) and apparently higher than normal-sham rats ( P ＜ 0. 05) ． In DM-control rats serum TNF-α values were significantly higher than those in normal-control rats ( P ＜ 0. 01) and apparently higher than those in DM-CLP rats ( P ＜ 0. 05) ; In normal-CLP rats peripheral blood neutrophil percentage was significantly higher than those in normal-control and normal-sham rats ( P ＜ 0. 01) ，and in DM-CLP rats peripheral blood neutrophil percentage significantly higher than those in DM-Control and DM-Sham( P ＜ 0. 01) ． Conclusion Inflammatory response of normal rats and DM rats to sepsis is different． This may be related to the difference of immune status in normal and DM rats．

Oxidative Stress Ｒesponse to 137Cs Ｒadiation in Ｒed Crucian Carps

2016, 33(02): 6-13.

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Abstract: Objective To study the oxidative stress response and oxidative stress biomarkers in red crucian carp following 137Cs radiation． Method The red crucian carps were irradiated with 0 Gy，1. 94 Gy，3. 88 Gy，7. 76 Gy，15. 53 Gy 137Cs，respectively，and the liver and gonad were collected 24 h，48 h，72 h，96 h after irradiation，respectively． SOD and GSH-PX activity were detected by commercially available kit; the liver，kidney，heart，brain were collected 7d and 14d after irradiation，respectively，and HSP70 content was detected by western blot．Ｒesult 1. 94 Gy to 15. 53 Gy137 Cs irradiation could induce the oxidative stress and could change SOD and GSH-PX activity，and HSP70 content in red crucian carp． 1. 94 Gy137Cs irradiation induced hepatic SOD activity increased，the SOD activity increased with the prolongation of observation time． 3. 88 Gy，7. 76 Gy，15. 53 Gy137Cs irradiation induced hepatic SOD activity decreased，and the SOD activity decreased gradually with the increase of 137Cs radiation dose． 137Cs irradiation at more than 1. 94 Gy doses induced gonadal SOD activity decreased，and gonadal SOD activity gradually decreased significantly with the increase of 137Cs radiation dose and with the prolongation of observation time． The GSH-PX activity in the red crucian carp liver and gonad showed little change in each 137Cs exposed group，which showed the trend of decrease with the increase of radiation dose． At 7 d and 14 d after 137Cs irradiation，the HSP70 expressions in liver，kidney，heart and brain of red crucian carps in each 137Cs exposed group were higher than that in control group． The HSP70 expression increased with the increase of 137Cs radiation dose and with the prolongation of observation time． There was some " dose-time-effect”relationship．Conclusion The gonad of red crucian carps was more sensitive to the oxidative stress response of 137Cs radiation，and the SOD in gonad and HSP70 in liver or kidney could be used as oxidative stress biomarkers．

Improving Effects of Hyperlipidemia of Different Contents of EPA and DHA in Fish Oil in Ｒats

2016, 33(02): 14-19.

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Abstract: Objective To observe the therapeutic effect of different contents of EPA and DHA in fish oil on Hyperlipidemia of rats． Method 70 male SD rats were fed with high-fat diet for 10 days，then randomly divided into seven groups according to TC levels: disease model control group，Fenofibrate group ( 28 mg /kg) ，Seal Oil group ( 333 mg /kg) ，fish oil A group ( 167 mg /kg) ，fish oil C group ( 150 mg /kg) ，fish oil D group ( 183 mg /kg) ，fish oil E group ( 167 mg /kg) ，ten rats in each group，respectively．Ｒats were treated with high-fat diet and drugs for another 30 days． Then the levels of TC，TG，HDLC，LDLC，IL-2，IL-6，the ratio of liver weight and body weight，and histopathology changes of the liver were examined．Ｒesult Compared to the disease model group Fenofibrate can decrease the level of TC，TG，HDLC，LDLC， the degree of hepatic steatosis of Hyperlipidemia of rats，but increase the ratio of liver weight and body weight ( P ＜ 0. 05 or P ＜ 0. 01) ; Seal Oil decreased the level of TC，TG，HDLC，LDLC( P ＜ 0. 01) ; fish oil A and fish oil E decreased the level of TC，HDLC，LDLC( P ＜ 0. 01) ; fish oil D decreased the level of TC，TG，HDLC，LDLC( P ＜ 0. 01) ; fish oil C did not decrease the level of TC，TG，HDLC，LDLC( P ＞ 0. 05) ; all of them did not decrease the degree of hepatic steatosis of Hyperlipidemia of rats． Conclusion Fish oil A，fish oil D and fish oil E can improve Hyperlipidemia of rats model to a certain extent．

A Investigation on the Safety of Clinical Application of Levocarnitine Injection Agent

2016, 33(02): 20-24.

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Abstract: Objective In order to investigate the safety of clinical application of Levocarnitine Injection ( 5 mL: 1. 0 g) ． Method Ear vein injection was used to evaluate the vascular irritation of rabbits，using left and right side of the same animals for comparison． The rabbit erythrocyte suspension was incubated with the test sample and then observed the hemolysis． Levocarnitine injection agent ( 5 mL: 1. 0 g) at 190 mg /kg and 95 mg /kg three times was gaven to the rabbits for sensitization，respectively． The symptoms of allergic reaction were not found when the sensitization concentrations were stimulated by 2 times volume，compared with a parallel reference in the same dose and concentration; Levocarnitine Injection ( 5 mL: 1. 0 g) in the high dose 190 mg / kg and a low dose of 95 mg /kg for passive skin allergy test．Ｒesult Levocarnitine Injection on rabbit red blood cell suspension ( in vitro method) and vascular stimulation was similar to the parallel control，with no hemolysis，no significant vascular stimulation． The symptoms of allergic reaction were not occurred in guinea pigs in the active allergic test．，Allergic reaction was negative． Conclusion Levocarnitine Injection ( 5 mL: 1. 0 g ) of clinical medicine meet the requirements of injection drug and can be used the base of safety evaluation．

Construction of Lentiviral Vector Overexpressing SV40LT

2016, 33(02): 25-28.

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Abstract: Objective To construct a lentiviral vector overexpressing of SV40LT，which serve a base to establish the immortalization of embryonic fibroblasts． Method SV40LT gene was obtained from 293T cells and then inserted into pLenti-GFP vector to construct recombinant plasmid pLenti-GFP-SV40LT，which identified by nucleotide sequencing． pLenti-GFP-SV40LT， pMD2. 0G， psPAX2 were co-transfected into 293T cells for lentivirus packaging．Ｒesult The Construction of Lentiviral Vector Over-expressing SV40LT was constructed successfully．Conclusion The recombinant lentivirus carrying SV40LT gene was constructed successfully，which lay a foundation to study immortalization of embryonic fibroblasts．

Development and Application of TaqMan MGB Probe real-time Fluorescence Quantitative PCＲ for Ｒapid Detection of Ectromelia virus

2016, 33(02): 29-36.

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Abstract: Objective To develop a TaqManminor groove binder( MGB) probe-based，sensitive and specific realtime fluorescence quantitative polymerase chain reaction ( qPCＲ) assay for rapid detection of Ectromelia virus ( ECTV) ． Method Primers and probes specific to HA( hemagglutinin) gene of Ectromelia virus were designed．Plasmid containing the sequence of HA gene was constructed as ECTV-DNA standard for quantitative analysis． A TaqMan MGB probeqPCＲ assay was developed． The sensitivity，specificity and stability of the assay were assessed．Then，the developed TaqMan MGB probe qPCＲ assay was applied to detect Ectromelia virus inclinical specimens．Ｒesult The developed TaqMan MGB probe qPCＲ assay was shown to be 100% specific to test a panel of fourteen organisms consisting of ECTV，other orthopoxvirus species，non-orthopoxvirus species，bacteria，fungi，parasites and cells． The technology was demonstrated to be highly sensitive，allowing a precise ECTV DNA quantitation over a range of ten orders of magnitude ( from 100 to 109 copies of standard DNA) ，and the limit of detection ( LOD) of the assay was determined to be 4 copies of target DNA． The reproducibility of this method was excellent as the Coefficient of Variation ( CV) for each point was less than 3%． Measurements of linearity such as Correlation coefficient ( Ｒ2 ) ，slope ( S) and efficiency ( Eff%) did not vary significantly among the test suggesting good accuracy and precision． The TaqMan MGB probe qPCＲ assay was successfully applied to quantifiable detection of viral genomic load in clinical specimens，and confirmed by using conventional PCＲ and sequence analysis． The assay described in this report generates complete result in 2 h and can be used as a fast diagnostic method．Conclusion This newly developed TaqMan MGB probe real-time fluorescence quantitative PCＲ method，has the characteristics of quick and easy，strong specificity and high sensitivity，suitable for Ectromelia virus detection in animal origin products，food and drug safety inspection，environmental monitoring and epidemiology investigation． It provides a specific and effective evaluation tool for rapid and quantitative detection of Ectromelia virus in clinical specimens，is worthy of popularization and application．

Heart Ｒate Analysis of the Guide Dogs in Active Level Test

2016, 33(02): 37-42.

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Abstract: Objective The aim of this study was to identify the law about heart rate changes of guide dogs in Passive test ( PT) ，to provide scientific information for early identify of dogs best suited to guide dog work． Method The subjects were 66 dogs from Chinese guide dog training center of Dalian． The dogs were tested using the PT before training and the behaviors were video-recorded． After training，37 failed guide dogs which were not suitable for guide dogs were determined by China guide dog training center． The heart rate reactions were recorded by ＲS 800CX Polar-Systems．Ｒesult The PT permits us to study active level of dogs． In the PT，the average score of guide dogs( 2. 0 ± 0. 11) was significantly different from the failed guide dogs( 3. 1 ± 0. 22) ，the guide dogs were less active than the failed guide dogs and there was no significant difference in mean values of heart rate changes between the guide dogs ( 19. 5 ± 8. 45) and the failed guide dogs ( 19. 6 ± 11. 35) ． In the dogs with lower active level ( score ＜ 3 ) ，the rest heart rate in guide dogs was significantly higher than that in failed guide dogs．Conclusion This study indicated that potential guide dogs have lower active level and higher rest heart rate compared to failed guide dogs，Heart rate reactions associated with PT can provide valuable information for early screening of guide dogs．

Ｒesearches of SPF Ｒats Alopecia Cases Caused by Feed Effects

2016, 33(02): 43-45.

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Abstract: Objective To discuss the reason which susceptibility to alopecia for SPF experimental rats in our center，and to provide the basis for scientific feeding management． Method Through the observation of the characteristics of SPF experimental rats with alopecia， sterilization effect and nutrients of the growth and reproduction group rat feed were detected． Combined with the results to improve the feed formulation，then gave the treat test and observe the effect．Ｒesult Vitamins and trace elements content change greatly in high-pressure feed．Use irradiated feed and intensive therapy as vitamins，hair growth results improved significantly． Conclusion The reason for alopecia of SPF experimental SD rats was determined to decrease of vitamins in feed during pulsating vacuum steam sterilization． It's more scientific to use irradiated feed in daily feeding．

Effect of the Combination of HSYA and β － boswellic Acid on Coagulation Function and the Whole Blood Viscosity in Blood-stasis Model Ｒats

2016, 33(02): 46-49.

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Abstract: To study the effect of HSYA and β-boswellic acid ( BA) on the whole blood viscosity and blood coagulation in blood-stasis model rats． 40 SD rats were randomly divided into control group ( constant volume of normal saline) ，model group ( constant volume of normal saline) ，HSYA group ( 100 mg /kg) ，β-BA group ( 200 mg /kg) ，drug combination group ( HSYA 50 mg /kg + β-BA 100 mg /kg ) ．Ｒats were given relevant medicine intragastrically every 12 h for consecutive 7 times． After the fifth administration， the model rats were given adrenalin hydrochloride ( 0. 7 mg /kg) subcutaneously twice with an interval of 4h． During the interval between two injections， ice-cold water was used to induce acute blood stasis model． Blood samples were collected within 30 min after the last administration to detect the coagulation parameters and the whole blood viscosity at all shear rates．Compared with control group， the whole blood viscosity at all shear rates was increased，and the thrombin time ( TT) ，prothrombin time( PT) and activated partial thromboplastin time( APTT) were all shortened in model group; the content of fibrinogen content( FIB) was increased; there was statistical significance( P ＜ 0. 05) ． Compared with model group，TT and APTT were prolonged and the content of FIB and the whole blood viscosity at all shear rates decreased in HSYA group，β-BA group and drug combination group; there was statistically significant( P ＜ 0. 05) ．The effect of drug combination was more obvious than single drug． HSYA and β-boswellic acid，and significantly improve coagulation function and the whole blood viscosity at all shear rates in blood-stasis model rats．

Advances on Music and Laboratory Animal Welfare

2016, 33(02): 50-53.

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Abstract: Music therapy is currently adopted and performed in the field of clinic medicine，psychological treatment and special education for young children with autism． The effects of music on physiology，cognition and brain development of animals suggest the possibility of providing music as a means of improving the welfare of laboratory animals． However，applications of music to improve animal environment，such as stress relief and depression reduction，are much limited and greatly lag behind in China． Based on the large variety of music types and auditory difference among animals，the benefits of providing music to the laboratory animals depend on the type of music and the species of animals．

The Development and Application of Humane Endpoints Technology

2016, 33(02): 54-60.

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Abstract: Humane endpoints mean without effecting the experiment result judgement，a point-in-time was defined as endpoint for alleviating animal pain． The systematic studies on humane endpoints has carried out internationally．In China，“Guidancesuggestion of caring laboratory animals”enacted in 2006 contains a section on application humane endpoints． In view of our country had already begun to emphasize“humane endpoints”，this paper briefed international current situation and was expected to give researchers overall view idea on humane endpoints．

Welfare Technology for Laboratory Animal in Production and Experiment

2016, 33(02): 61-65.

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Abstract: Animal welfare is an important part of laboratory animal science，and has been paid more attention to with the development of laboratory animal science． In the review，we analyzed the problems which were often found in animal production and experiment，and also provided some solution and suggestion．

The Application of Image Technology in Animal Experiment

2016, 33(02): 66-69.

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Abstract: The image technology is to show the internal structure and density of the internal organs in the form of image via the interaction between the media and the body． Commonly imaging methods include X-ray，CT，MＲI，ultrasound，angiography and biological luminescence etc． It can be continuous，accurate，and non-invasive to observe the changes of diseases in animals by different image technology． By these methods the level of experimental research was enhanced greatly，and the pattern of disease detection was promoted from traditional anatomical morphology to molecular level development of functional，metabolic，and even receptor and gene．Which involves the welfare refers to ensure the animal’s well being during the whole image detection process． The purpose is to find a balance between animal welfare and human interests． That is concerned about the factors that may affect the animal＇s well being，and less of an animal＇s pain in the process of image． At the same time，when the welfare conditions are satisfied with animal well being the effectiveness of image experiments can be maximized．

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