Table of Content

28 August 2019, Volume 36 Issue 04

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High Fat and High Sugar Diet Induced the Fat Model of Bama Miniature Pigs

2019, 36(04):  1. 

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Objective The effects of high fat and high sugar diet(HFHSD) on fat distribution and blood biochemical parameters in Bama miniature pigs. Method Fourteen Bama minipigs were divided into two groups. Experimental group pigs fed with high fat and high sugar diet for 30 weeks.Six Bama miniature pigs in control group were fed with normal feed. After 30 weeks, body mass index, fat distribution and biochemical indexes of blood were measured. Result The body weight, body mass index (BMI) and total fat content especially visceral fat content increased significantly in experimental group.Blood biochemical indicators of insulin, total cholesterol, high and low density lipoprotein cholesterol, blood creatinine, uric acid and urea were increased significantly compared with control group. Conclusion High fat and high sugar diet can cause fat accumulation in Bama miniature pigs, especially visceral fat.Blood biochemistry is characterized by hyperlipidemia and impaired renal function.

Expression of Bax Gene in Cerebellum of Rats and Psychodependence Drug Addiction Model Copied

2019, 36(04):  6. 

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Objective To study the effects of psychotropic drug (heroin) on the expression of Bax gene in the cerebellar of addicted rats. Method An addictive rat model was used to establish with a dose-increasing method and the gene expression of Bax in the cerebellar cells was detected by ELISA. Result After injecting heroin for 7 days, the rats showed obvious withdrawal symptoms. The levels of Bax, Caspase-3, and Caspase-9 in cerebellar cells were increased 88.21%, 94.21%, and 87.24% respectively, which was significantly different compared with the control group (P<0.01). Conclusion The damage effect of psychoactive drugs(heroin) on brain tissue cells may be achieved by inducing the expression of cerebellar Bax gene and activating caspase.

The Changes of Plasma and Tissue Levels of Sex Hormones in Nude Female Mice with Green Fluorescence at Different Stages

2019, 36(04):  10. 

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To understand the changes in plasma levels of estrogen, progesterone, prolactin, and prolactin receptor mRNA in mice during pregnancy and lactation. Plasma levels of progesterone were measured by electro-chemiluminescence and relative mRNA levels of prolactin and prolactin receptor were measured by fluorescence quantitative PCR. During the gestation period, estrogen and prolactin levels decreased continuously, progesterone levels increased first and then decreased slowly, prolactin mRNA levels were low in tissues, and prolactin receptor levels were high.During lactation, the levels of estrogen and progesterone are relatively stable, the levels of prolactin mRNA in tissues are relatively high, and the levels of prolactin receptors are relatively low. Different levels of sex hormones in nude mice with green fluorescence were different in different periods, and they participated in the process of pregnancy regulation in nude mice. It provides basic reference value and endocrine level for solving the problems of female mouse infertility and abortion.

Genetic Analysis of BALB/c Mice by Using Microsatelite DNA Markers and Biochemical Markers

2019, 36(04):  14. 

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Microsatelite DNA marker is one of the most valuable genetic marker, in order to discuss it’s applicability in genetic monitor of BALB/c mice, and lay the foundation for establishing STR genetic quality monitor method, 20 microsatelite DNA markers and capillary electrophoresis were adopted to genetic detect BALB/c mice, and compared with biochemical marker analysis in current national standards．The result showed that the DNA fragments of 20 microsatelite loci of each sample were the same, there were no new alleles. Biochemical marker analysis also showed that fourteen biochemical marker genes were homozygous, the phenotypes of biochemical markers were consistent, according to current national standards, the sample conformed to strain characteristics. The STR method result were coincident with that of Biochemical marker analysis. The microsatelite DNA marker method is more accurate, reliable, rapid and convenient, and the 20 microsatelite DNA markers selected in this study could be used to monitor genetic quality of BALB/c mice．

Establishment and evaluation of a model of osteoarthritis in the rabbit knee

2019, 36(04):  18. 

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Objective To establish and evaluate a rabbit model of knee osteoarthritis induced by articular cartilage defect. Method The animals were randomly divided into 3 group, the sham group, the model group and the positive control glucosamine group, each group of 12 animals. After 4 weeks, the drugs were administrated intragastrically once daily for 4 weeks. After the last administration, the animals were executed. ELISA kits were used to detect the level of IL-1β and TNF-α in the serum and joint fluid. The articular cartilage and synovial membrane were stained by HE staining. Mankin’s score was used to assess articular cartilage damage. Result Compared with the sham group, the level of IL-1β and TNF-α in the serum and joint fluid of the model group was significantly increased, while that of glucosamine decreased the level of these inflammatory cytokines. HE staining showed the structural changes and defects of cartilage, and the synovial membrane was infiltrated by inflammatory cells in the model group. The score of cartilage was significantly higher than that of the sham group. Glucosamine can reduce cartilage defects, reduce the infiltration of synovitis cells and reduce the score of cartilage. Conclusion Articular cartilage drilling is able to establish the rabbit model of knee osteoarthritis, reflect the main features of osteoarthritis. The injury and inflammation induced by the model can be reduced by administrating with positive drug. Therefore, this model can be used for the evaluation and screening of arthritis drugs.

Effects of DHEA on Superovulation and Oocyte Quality of C57BL/6J Mice.

2019, 36(04):  22. 

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Objective To investigate the effect and mechanism of DHEA on the quality of oocytes of C57BL/6J mice after superovulation. Method Thirty C57BL/6J mice at the age of 3 weeks were randomly divided into 3 groups, which were control group、low does group(2 mg/kg·d-1)and high does group(5 mg/kg·d-1), respectively. DHEA was administrated via the gastrointestinal tract for one week. Oocytes were collected after superovulation and underwent IVF. qRT- PCR was used to detect gene expression of Grem1、Has2、Ptgs1、Ptgs2 and Vcan，the makers of cumulus cells collected after superovulation. Gene expression of Caspase3 and Bcl2l10 were also detected by qRT-PCR. Result After DHEA treated, IVF showed that the proportion of heterotypic eggs decreased in the experimental group. The transcription levels of Has2, Ptsg1 and Vcan in cumulus cells of experimental group were up-regulated compared with those of control group, while the mRNA levels of Germ1 and Ptgs2 were down-regulated. The mRNA level of Caspase3 was down regulated and the Bcl2l10 mRNA level was unregulated. Conclusion After DHEA treatment, the apoptosis of cumulus cells can be inhibited, and the quality of oocytes can be improved.

Establishment of Hepatic Fibrosis Model Induced by CCl4 in Inbred C57BL/6 Mice

2019, 36(04):  28. 

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Objective Compared the effects of different doses of carbon tetrachloride induced liver fibrosis in inbred C57BL/6 mice, in order to established a stable liver fibrosis model for the study of liver diseases and related drug mechanisms. Method The 5-week-old C57BL/6 mice were subjected to high, medium and low dose by CCl4, after the experiment collected blood and liver tissue to detect the levels of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and liver pathology. Result After the induction of inbred C57BL/6 mouse drug, the medium and low dose group could survive for 8 weeks, the increase of serum transaminase content and the pathological changes of liver tissues had a dose-time effect, and the mice with severe pathological changes had a large area of liver tissue necrosis and were wrapped by fibrous tissue. Conclusion Eight weeks after administration of 10% carbon tetrachloride, inbred C57BL/6 mice were induced to form a stable mouse model of liver fiber.

Establishment and Analysis of Hyperlipidemia Model in Cynomolgus Monkey

2019, 36(04):  31. 

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Objective To establish Cynomolgus monkey model of hyperlipidemia. Method Cynomolgus were feed by high fat diet,the levels of total cholesterol(TC), high-density lipoprotein(HDL), low density lipoprotein (LDL), triglyceride(TG) of the Cynomolgus monkey were determined at 4, 8, 12, 18months for analyzing the trend of each paramaters above. Result When it was 4 months, the level of TC,HDL,LDL were higher significantly than that before feeding the high fat diet(P<0.01). When it was 8 months, the level of TG was higher sifnificantly than that before feeding the high diet(P<0.01). And the levels of TC, HDL, LDL and TG of model group were significantly different from that of the control group(P<0.01) at 8 months, 18 months. At this experiment, TC, HDL,LDL,TG all showed a increase trend, expecially the levels of TC and LDL much higher than the normal levels of Cynomolgus monkey. Conclusion Cynomolgus monkey of the model group has the feature of high total cholesterol and high low-density lipoprotein cholesterol, the method to establish Cynomolgus monkey model of experimental hyperlipidemia was feasible.

Breeding and Genotype Identification of Ifnar Gene Knockout Mice

2019, 36(04):  35. 

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Objective To analyze the identification and breeding method of ifnar knockout mice, and provide an ideal animal model for virus further studying. Method The introduced homozygous ifnar gene knockout mice were bred and reproduced in one cage with one male and two female mice together. The offspring mice genomic DNA was extracted from mice tail and amplified by PCR, and PCR products were analyzed by agarose gel electrophoresis for genotype identification. Result The homozygous ifnar knockout mice were breeding successfully in our facility,and the genotype can be identified by our established PCR method .

Study on Preparation of Catalase-2 Samples Used for Proficiency Testing and Measurement for the Stability and Uniformity

2019, 36(04):  39. 

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Objective To prepare and evaluate catalase-2 sample(a type, b type)with good stability and uniformity. Method Extracting 4% samples randomly to finish the uniformity test. Samples were also extracted in the freeze thawing stability test for at most 4 times. As for the time-temperature uniformity, we used time(day) and temperature(℃) as the coordinate value: 1, 2, 3, 7,15, 30 days were used as time coordinate, and 4, 20, 37℃ as temperature coordinate; and 2 bottles of each type samples were observed at every fixed point. Result A type and b type clearly performed a and b band on their own., and all of them were able to be stable after 4-time freeze thawing. And also the time-temperature stability test showed that standard samples of a type and b type could be present stable for 30 days at 4,20℃ and 37℃. Conclusion Catalase-2 samples behaved well in the uniformity and stability test, which were up for the requirements of proficiency testing.

Association between MHC Gene Polymorphism and VMC

2019, 36(04):  44. 

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Objective To investigate the correlations between MHC gene polymorphism and the development of viral myocarditis and to identify the susceptibility genes for VMC for further elucidating the pathogenesis of VMC and providing the guidance for researches on further gene therapy for VMC. Method Few genotypes of H2-Eb(including MudoEb 5、MudoEb7) in 32 VMC mice and 32 normal control group were detected by using polymerase chain reaction, as the same time,the serum level of IFN-γ and the pathological changes of VMC mice were monitored to reflect the relationship between VMC and MHC gene polymorphism. Result The frequencies of H2-Eb MudoEb5/Eb7 in VMC mice(37.5%/9.4%) were higher than those in normal control group(14.1%/3.1%)(MudoEb 5 λ2=20.33 P=0.00 OR=13, MudoEb 7，λ2=5.38 P=0.02 OR=5.7). Conclusion The genotypes MudoEb5、MudoEb7 might be the susceptibility genes for VMC in mice.

Effect of Puffing on Nutritional Components of Experimental Dogs

2019, 36(04):  51. 

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Objective Dogs are one of the earliest domesticated animals of human beings. In the 1940a,dogs began to be used as experimental animals for research. They are mainly used in experimental surgery, basic medicine, pharmacology, toxicology and some diseases in biomedicine, which is of great significance to the development of biomedicine. Providing standardized formula feed for experimental dogs is an important step to improve the scientific nature of experiments. At present, puffing technology is used in the production of experimental dog food. This paper mainly studied the changes of protein, fat, amino acid, vitamin and trace elements in dog food before and after puffing. Method According to the Chinese Feed Composition and Nutrition Value Table (28th edition), the nutrient distribution formulas of experimental dog food were calculated, and then the finished experimental dog food was sent to the laboratory to test the actual value, and the two were compared. Result After puffing, fat loss was 6%, histidine, lysine, methionine loss was 8%, 6%, 2%, vitamin E, vitamin B2, nicotinic acid, choline, vitamin B6 loss rate was 29%, 65%, 46%, 1%, 81%, respectively. Conclusion Under the expansion parameters set in this study, the nutrient losses in the experimental dogs food are as shown in the result . The effect of extrusion parameters on nutrients is different. Therefore, it is suggested that peers can adjust dietary formulation appropriately according to actual production situation.

Comparative Study of Small Animal Micro-CT, Ultrasound and Aortic Intubation in Evaluating Left Ventricular Function

2019, 36(04):  55. 

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Objective To study the consistency and correlation of micro-CT, aortic intubation and echocardiography in measuring left ventricular function parameters, and to evaluate the value of micro-CT in evaluating left ventricular function. Method Cardiac function tests were performed on 10 pairs of normal and heart failure rats by micro-CT, aortic intubation, and ultrasound. Left ventricular function parameters were measured using cardiac function analysis software: end-systolic volume, end-diastolic volume, and ejection fraction. The micro-CT result were compared with the result of aortic intubation and echocardiography. Result All the rats were examined by micro CT, aortic catheterization and echocardiography. There was a good correlation between the parameters of left ventricular function measured by micro CT, aortic catheterization and echocardiography. Conclusion micro-CT can be used to evaluate left ventricular function and is a feasible solution for accurate measurement of ventricular function.

Comparison of the Viscera Coefficient of CMU/1 and CMU/2 Inbred Strains of Gerbil

2019, 36(04):  60. 

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Objective To determine the body weight and main organ weight of inbred CMU/1 and CMU/2 inbred Mongolian gerbils, and to explore and establish the biological index system of main organs of Mongolian gerbils. It provides the necessary parameters for animal experiments. Method The body weight and organ weight of the 3～4 month-old gerbil in the inbred line were analyzed statistically. Result The difference between lung coefficient CMU/1 and CMU/2 inbred gerbils significantly, other no significant difference; Kendall coefficient (W) close to 1, the main organs of the overall development of better coordination; linear regression equation, the CMU/1 P<0.05,CMU/2 in the liver, thymus, lung P<0.05, two of the sample had a linear relationship between. The thymus of the liver, spleen, adrenal gland, ovary, pancreas and CMU/2 of CMU/1 was positively correlated with body weight. Conclusion There is significant difference in lung organ coefficient between CMU/1 and CMU/2 inbred strains of gerbil, and there is no obvious influence on the organ coefficient of the gerbil in the inbred line.

The Effect of Western Diet on the Apolipoprotein E Gene Knock out Mice

2019, 36(04):  63. 

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Objective To study the effect of western diet and standard diet on the blood fat and aortic arch pathology of apolipoprotein E gene knock out mice. Method We feed 3 months old ApoE-/- mice with western dietary or standard diet for 3 months, 4 months and 5 months, respectively, then collect peripheral blood and aortic arch. We use Automatic blood analyzer to detect lipid level in serum, and observe the formation of atherosclerotic plaque through sectioning and staining.Result On the blood lipids detection, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) in serum of mice feeding western diet and standard diet were significantly higher than the control group. In addition to HDL-C mice 4 months feeding, the lipid index of western diet lipid groups are significantly higher than the standard diet group on the same age. On the stained sections, western diet aortic plaque and incidence of atherosclerosis are significantly increased.Conclusion Both Western dietary and standard diet can induce ApoE-/- mice showed AS symptoms, the effect of western dietary is more obvious.

A New Method is Simple and Easy to Extract the Cerebrospinal Fluid in Mice

2019, 36(04):  68. 

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Objective To establish a simple method for drawing out the CSF of mice and provide references for relative researches. Method First place the mouse in the position that the head forms 120°, shave the skin and separate the subcutaneous tissue and muscle bluntly. As a result , the dura mater of the cisterna magna appears clearly. Penetrate the white dura mater and then reach the cerebellomedullary cistern by the capillary tube. Obtain the CSF and withdraw the tube. Finally, suture the mouse. Result 2—5 μL CSF samples can be collected successfully with mice survived. Conclusion Compared with others, the method we established is proved to be with cheaper material, higher success rate and survival rate.

Analysis of Heat and Mass Transfer inside the Individual Ventilated Cage

2019, 36(04):  71. 

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Objective To effectively control the environmental comfort of in the individual ventilated cages (IVC), to ensure the survival and health of the experimental animals in the cage, to test the temperature, humidity and ammonia concentration in the cage, and to analyze the heat and mass transfer mechanism in the cage, and provide reference for the control of living environment (closed building space and equipment) of experimental animals. Method Seventy-five mice were randomly divided into 5 groups, and each group was randomly divided into 5 groups according to the number of 1 to 5 increments. They were kept in the same IVC equipment, and the temperature, humidity and ammonia concentration in the cage were tested for 7 consecutive days. Result With the increase of the number of days, the temperature difference between the cage and the room was basically the same, the relative humidity difference and the ammonia concentration difference gradually increased. At the same time, the ammonia concentration difference inside and outside the cage was significantly different from the number and duration of the animals (P<0.05), if the case have more animals and feed longer time, the greater the difference in ammonia concentration. Conclusion The temperature inside the IVC cage is greatly affected by the temperature in the facility. The relative humidity is mainly related to the relative humidity of the supply air and the number of air changes. The ammonia concentration is related to the number of air changes and the number of animals.

Discussion of the Management for “three Wastes” Generated by Laboratory Animal Facility

2019, 36(04):  77. 

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The laboratory animal facility is the sum of the buildings and equipment used for the production and research experiments of laboratory animals. The waste materials, waste water and waste gas (called “three wastes”) generated during the process of laboratory animal facility will pollute the environment, however, the mechanism of reasonable and effective management for the “three wastes” is still not developed yet. In this paper, we will discuss the problems involved in the management for “three wastes” of the laboratory animal facility, in order to provide a feasible plan for establishing a reasonable “three wastes” management mechanism.

Research Progress of Genetic Diversity in Cat (feliscatus)

2019, 36(04):  81. 

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Since the late 19th century, Cats (feliscatus) has been used for scientific research. Now, it is widely applied in various fields of medicine. In addition, cats are gaining more and more attention as animal models of human genetic and infectious diseases. The quality of cats (especially genetic quality) has a significant impact on the accuracy, reproducibility, and scientificity of the findings. It is comparatively vital to carry out genetic quality testing on a regular basis to avoid the deterioration of germplasm, genetic drift and the error of experimental result . With the advent of biochemical markers and DNA molecular markers, a more simple and reliable method for understanding the genetic structure of cats has been provided. In this paper, the current research status and existing problems of biochemical markers and DNA molecular markers in the study of cat genetic structure and polymorphism are discussed.

Humanized Mouse Model Reconstructing Human Immune System and Its Application in Study of Blood System Diseases

2019, 36(04):  86. 

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Animal models play an important role in the study of human disease pathogenesis and experimental study of pharmacology. Mice are currently the main experimental animals，However, it is different from the human background,and it cannot fully simulate the human immune system and disease state. Researchers can build humanized mouse models of the immune system and humanized mouse disease models by implanting human cells or tissues into immunodeficient mice. Humanized mice can obtain a mouse model with a high degree of humanization of the immune system by optimizing conditions such as the source of human hematopoietic stem cells and other conditions. At present, the humanized mouse model of GVHD and the humanized mouse model of leukemia have been used in the study of disease pathogenesis and the development of new clinical therapeutic drugs.