Table of Content

28 April 2018, Volume 35 Issue 02

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Preliminary Study on the Biological Characteristics of the Breeding Population of the Inbred Strain of the Diabetic Model of Gerbil

2018, 35(02):  1. 

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Objective The basic data of genetic biochemical markers and biological characteristics of the breeding population of the inbred strain of Mongolian gerbil diabetes model. Method Using Beckman automatic biochemical analyzer (CX5) and Japan photoelectric automatic blood cell analyzer (MEK-7222 K) of 17 blood biochemical and 22 blood physiological indexes were determined; detection method of electrophoresis acetate fiber 26 biochemical marker; F20 rat, inbred animal production in accordance with the way of breeding, analysis of fetal the interval, number, weaning number statistics, growth curve. Result The result of the blood physiological and biochemical comparison between ALP, HDL-C and CRE index were significant differences (P<0.01); the difference of MCV, MCH, EOS, GLU, TG, LDH were significantly (P<0.05); a single genetic biochemical marker; birth weight is 2.68—3.38 g, the survival rate of weaning 68.31%, fetal long time. The quality of the 3—6 week old body weight increased rapidly. Conclusion The result of the study provide important reference materials for the identification of their lines and the application of biomedical applications.

Effect of Endoplasmic Reticulum Stress in Rat Congestive Heart Failure Induced by Myocardial Infraction

2018, 35(02):  6. 

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Objective To explore the effect of endoplasmic reticulum stress in rat congestive heart failure induced by myocardial infraction. Method The rat model of heart failure was established by ligating the left anterior descending coronary to produce acute myocardial infarction．The rats were divided into 5 groups：sham group and groups of post-operation at time points of 2, 4, 6 or 8 weeks, respectively. The ventricular functions were assessed by hemodynamic.The expression of GRP78, CHOP, caspase-12, caspase-3,and Bcl-2 were detected by Q-PCR. Result Left ventricular systolic pressure (LVSP) and maximal rise and decline velocity of left ventricular pressure decreased (P≤0.05 or P≤0.01), but left ventricular end diastolic pressure (LVEDP) increased (P≤0.05) in model group compared with sham group. The expression of endoplasmic reticulum chaperones GRP78, CHOP, caspase-12 and caspase-3 was significantly increased (P≤0.05 or P≤0.01) in model group compared with sham group. And the expression were positively correlated (P≤ 0.01)with the feeding time of post-operation. The expression of Bcl-2 was reduced gradually (P≤0.01) in model group compared with sham group;And the expression were negatively correlated (P≤0.01)with the feeding time of post-operation. Conclusion The congestive heart failure induced by myocardial infraction is associated with endoplasmic reticulum stress. It suggest that endoplasmic reticulum stress-induced apoptosis is a possible mechanism of heart failure effecting on promoting.

Expression Pattern of Scn3b in Multiple Tissues from Hereditary Diabetic Mongolian Gerbils

2018, 35(02):  11. 

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Objective To investigate the mRNA and protein expression level of sodium channel beta subunit 3 gene(Scn3b) in 4 tissues of spontaneous hereditary diabetic and non-diabetic gerbils. Method We selected 6 diabetic and 6 non-diabetic control gerbils, and collected liver, kidney, skeletal muscle and brain from each animal, respectively. The mRNA and protein expression of Scn3b was evaluated by Real-time PCR and Western blotting in diabetic and non-diabetic control groups. Result Compared with control group, mRNA and protein expression of Scn3b reduced in liver and brain from diabetic gerbils, and the difference in liver between the two groups was statistical significant. Conclusion Scn3b expression decreased in liver and brain in diabetic gerbils, suggesting that Scn3b in liver and brain may be involved in development of gerbil diabetes.

Analyzsis of Population Genetic Structure of Rongshui Miniature Pig by STR Scan Technology

2018, 35(02):  16. 

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To evaluate the genetic structure of Rongshui miniature pig by STR scan technology. Twenty five microsatellite sites selected Beijing local standards were applied to amplify the genomic DNA from 30 Rongshui miniature pig for F1 generation in the miniature pig base of Guangdong medical laboratory animal center by PCR, the PCR product were analyze by STR scanning, software popgene 32 was used to analyze the genetic structure. For the Rongshui miniature pig population for F1 generation, the average observed number of alleles is 9.4400, the average effective number of alleles is 5.2966, the average shannon index is 1.8085, and the average effective heterozygosity is 0.7737. The STR scan technology can be application in genetic structure analysis of miniature pig population, and we initially grasp the genetic structure of Rongshui miniature pig

Effects of Glycerol-3-phosphate on Humoral Immune Response Induced by Hepatitis a Virus Vaccine in Mice

2018, 35(02):  21. 

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Objective To investigate the effect of G3P(Glycerol-3-phosphate) on humoral immune response induced by hepatitis a virus vaccine in mice. Method Forty nine ICR female mice were divided randomly into 7 groups (7 mice in each group): negative control group, antigen group, aluminum adjuvant group and 4 groups of G3P. The specific anti-HAV IgG levels in the serum were detected by enzyme-linked immunosorbent assay (ELISA) at the 4th, 8th, 12th, 16th, and 20th week after immunization. Result The serum anti-HAV IgG levels were detected in antigen group, aluminum adjuvant group and 4 groups of G3P at the 4th, 8th, 12th, 16th, and 20th week after immunization, and the serum IgG level of the mice increased first and then decreased, and the level reached the highest at 8th week. The serum IgG level of G3P group was higher than the antigen group and the difference was statistically significant (P<0.05)at 4th week and 8th week, besides, the serum IgG level of G3P group was higher than aluminum adjuvant group at 4th week and the difference was statistically significant(P<0.05).The serum IgG level of G3P 2 mg was the highest in all of G3P,and the level was higher than the antigen group at 4th, 8th, 12th, 16th, and 20th week after immunization and reached the peak at the 8th week, with the value of lg(3.064 ± 0.07851), aluminum adjuvant also reached the peak at 8th weeks, with the value of lg (3.150 ± 0.1382). Conclusion G3P could significantly and rapidly enhance humoral immune response induced by HAV vaccine in mice; the result suggest it may be used to develop a new type of human vaccine adjuvant．

The Pathological Examination of Laboratory Rodents of Beijing in October 2017

2018, 35(02):  27. 

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Objective Though detecting the pathological changes of laboratory rodents in Beijing, it provided a basis for the quality monitoring and improvement of laboratory rodents in Beijing. Method We collected hearts, livers, spleens, lungs, kidneys and intestinal tracts from 400 laboratory rodents in Beijing including 180 rats, 80 mice, 120 guinea pigs and 20 golden hamsters, and then made the paraffin sections stained with HE. The samples of the hepatocellular swelling were also stained by PAS and oil red O staining made from frozen sections, and observed under the microscope. Result The lesions of laboratory rodents mainly were hepatocellular swelling of mice, rats, guinea pigs and glycogen deposition in different degree, and liver fatty degeneration in guinea pigs; In addition, the mice and golden hamsters, especially hamsters, showed more liver necrosis and inflammatory cell infiltration. And the heart of mice and guinea pigs were hyperplasia, inflammatory cells infiltrated of kidney could be seen in mice. Conclusion The vast majority of laboratory rodents were pathologically diagnosed as healthy animals. But the feeding and management of some rodents still need to be strengthened, to provide a reliable and effective premise for the accuracy of the research experiment.

Expression Level of Nf-κb in Multiple Tissues from Hereditary Diabetic Mongolian gerbils

2018, 35(02):  33. 

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Objective To investigate the mRNA and protein expression level of Nf-κb in 6 tissues of diabetic gerbils, and to explore the role and target tissue of Nf-κb in diabetic gerbil model. Method We selected 6 animals from diabetic and control group, respectively; and collected skeletal muscle, liver, adipose, kidney, heart and brain from each group. The expressionsat mRNA and protein level of Nf-κb were evaluated by Q-PCR and Western blotting in diabetic and control groups. Result Compared with control group, mRNA expression of Nf-κb had an increasing trend in skeletal muscles, kidney and brain of diabetic gerbil. The mRNA level of Nf-κb significantly decreased in adipose and showed decreasing tendency in liver and heart. After detecting protein expression, we found the protein level was consistent with mRNA in all tissues except liver. Conclusion Adipose tissue is the target tissue of Nf-κb in diabetic gerbil.

Methods for Concentration of Enteroviruses from Fecal ofTupaia belangeri: a Comparative Study

2018, 35(02):  38. 

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Objective To provide a simple and efficient method for concentration of the virus in feces of Tupaia belangeri, the sensitivity, different concentrations and recovery of three method for enterovirus concentration were observed. Method The centrifuged and filtered wild tree shrews stool supernatant was added indicative virus (tree shrew’s reovirus TRV), and the fecal supernatants were concentrated by PEG precipitation combined with ultrasonic separation, Fe3+-based virus width=11,height=14,dpi=110occulation and GBviroFast concentrated virus kit. To determine the effect of concentrating three method , real-time PCR technique was used. Result Load of TRV virus suspension is 1.34E+05; TRV load concentrated by Fe3+-based chemical flocculation is 1.86E+04, and the concentration efficiency is 49.57%. TRV load concentrated by GBviroFast concentrated virus kit is 0.98E+04,with concentrated efficiency of 26.12%. TRV load concentrated by PEG precipitation combined with ultrasonic separation is 2.30E+03,and the concentration efficiency only is 6.13%. Conclusion Three method can concentrate stool supernatant which contain low concentrations of enteroviruses, Fe-based chemical flocculation is the best, GBviroFast concentrated virus kit secondly, PEG combined with ultrasonic separation method is the worst.

Development of Liquid Chip Technology for the Detection of Marek’s Disease Virus

2018, 35(02):  42. 

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Objective In order to establish a double-antibody sandwich liquid chip technology assay for detection of Marek’s Disease Virus. Method The capture antibody was coupled to the 19 microsphere. We optimized the coupling capacity of capture antibodies, the working concentration of detection antibody and SA-PE, respectively. The specificity, sensitivity, reproducibility and clinical samples of this method were evaluated. Result The optimal coupling capacity of capture antibody was 2 μg of 1×106microspheres. The optimal working concentration of detection antibody and SA-PE was 2 μg/mL and 4 μg/mL, respectively. The method was specific for Marek’s Disease Virus, and there was not cross reaction with other viruses; The limit of detection reached 125 pfu/mL; Reproducibility analysis showed that the intra- and inter-assay coefficient of variation was 0.85% and 2.4%, respectively; The detection rate of 58 clinical samples was 31%. Compared with PCR, the total coincidence rate was 94.8%. Conclusion These result showed that the method was high specificity, sensitivity and reproducibility. The assay has important significance for monitoring, preventing and controlling Marek’s Disease, and could be useful for screening of specific-pathogen-free (SPF) chicken.

Establishment of Animal Models of Subacute Alcoholic Liver Injury in Rats

2018, 35(02):  47. 

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Objective To determine the most convenient conditions for the establishment of subacute alcoholic liver injury by comparing different doses of alcohol. Method The SPF female Wistar rats (150～180 g) were selected as the model group and control group. In the model group, the alcohol content was 15% and 30% respectively. The control group was given distilled water, with 15 mL/kg·BW, and the trial period was 8 weeks. At the end of the experiment, the serum glutamate aminotransferase (ALT), aspartate aminotransferase (AST) and ethanol dehydrogenase (ADH) were measured from orbital venous plexus of rats. Hepatic histopathological lesions were determined by liver tissue, and MDA, SOD, GSH, TG were measured. Result There was significant difference in MDA between 30% alcohol group and blank control group (P 0.05).Model group rats blood lipid metabolic disorder and the accumulation of fat in the liver could be founded and histologic examination showed diffuse hepatocellular fatty degeneration, formation of different size of the fat granule in the cell. According to the score of the liver disease scale, 15% alcohol, 30% alcohol group compared with blank control group, the degree of liver steatosis has significant difference (P< 0.01, P< 0.01). Conclusion Rats filled the stomach 8 weeks, using the ordinary liquor of alcohol content 30% can cause liver damage. The result can be repeated and subacute alcoholic liver injury animal model of rats is successfully established.

Application of Self-made Endotracheal Tube in Anesthetized Small Tail Han Sheep

2018, 35(02):  52. 

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Small Tail Han sheep have become an importmant experimental animal applying to etiology and treatment of related spinal column diseases because their changes similar with human. The trachea cannula could ensure the airway smooth, auxiliary breathing, easy to monitor and reduce intraoperative traction damage to the trachea in the sheep spinal surgery experiment.But It is more difficult for trachea cannula of sheep since its longer trachea and limited mouth opening.Because endotracheal tube in the market is relatively shorter, autheros succeed in making endotracheal tube for trachea cannula after many attempts. It is key to familiar with the anatomical structure of the sheep respiratory system as well as correct operations should be applied. Therefore in this article Small Tail Han sheep anesthesia, tracheal intubation, intraoperative monitoring and postoperative care were summarized in order to provide reference for the experimental staff.

Establish of Specific Pathogen-free Shedldrake duck Breed Populations

2018, 35(02):  56. 

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Objective Establish specific pathogen-free Anas platyrhyncha var.domestica breed populations. Method Highly quality breed species of Shaoxing Shedldrake No. I and Jinding duck breed eggs were based on as original populations, various biological purification measures were adopted, including eliminated specific avian diseases antibodies and antigen positive individuals, immunized with killed vaccines, raised in positive pressure high efficiency particulate air filter, fed with 60Co-treated feed, acidified water, strictly controlled the flow of people and logistics. Simultaneously, tests of molecular genetics, biological characteristics and disease susceptibility were carried out to standardize the ducks. Result HBK-SPF and SPF-JD duck breed populations with specific pathogens-free, clear genetics background and disease susceptible were successfully established. Conclusion The established specific pathogen-free duck breeds will enrich Chinese laboratory animals resources and provide important support for life science research and development of biological products.

Applications of Environmental Monitoring Mobile Platform in Laboratory Animal Facility

2018, 35(02):  60. 

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Objective In strict accordance with Chinese national standards requirements of environmental testing laboratory animal facility, while improving efficiency and saving labor costs. Method Design of a portable laboratory animal facility environment monitoring mobile platforms for environmental testing.The platform can take test points by adjusting the height, width and length in the experimental animal facility. Result The platform can be a good application in a variety of facilities to avoid systematic errors operator selected points. Conclusion The platform is broad prospect, but the convenience to be improved.

Husbandry of Laboratory Red Crucian in Aquarium

2018, 35(02):  65. 

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In order to improve the standardized content of laboratory red crucian carp,the aquarium suitable husbandry strategies for this kind of fish is proposed by summing up many years of practical experience and referring to the literature.Select the size of about 1200 mm × 480 mm × 755 mm (length × width × height) of the rectangular aquarium, set aeration pumps, filters, lighting and other kits, simulated pond habitat for scenery. After aeration 3 days long，the tap water is injected into the aquarium. The water quality and the environment of the aquarium were controlled goodly, maintaining the water temperature at 18-24 ℃, dissolved oxygen in ≥ 6 mg/L, pH value of pH7.5-8.5, light in the 100-200lx, hardness (CaCO3) in 95-105 mg/L, ammonia (NH3) concentration in less than 0.02 mg / L and so on. As an example of 100 mm length fish, according to the density of water per cubic meter with 80 tail fish, put the laboratory red crucian carp into the aquarium. Feeding the full-price pellets daily for 1 or 2 times, replacing part of the water every 3-7 days for 1 time, replacing all of the water for 14-21 d and cleaning the aquarium once, insisting on daily management and cleaning and disinfection. The technical method described in this article is a husbandry technical strategy suitable for laboratory red crucian carp in aquarium.

Application of Western Blot in the Study of Myocardial Ischemia in Rat Inflammatory Cytokines

2018, 35(02):  65. 

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Objective To improve the success rate of each experiment. Method Western blot technique. Result Western blot of the rat myocardial inflammatory cytokine has its particularity. The success rate was greatly increased. Conclusion These experience or experimental considerations were related to success or failure of each experiment. Inflammatory cytokines(IL-6, IL-1β, TNF-α and NF-κB)involved in the pathophysiological process of myocardial ischemia.

The MXH10/Mo-lpr/lpr Mice’s Application in Cancer Diagnosis and Treatment

2018, 35(02):  74. 

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This review introduces the MHX10/Mo-lpr/lpr mice that it shows the systemic lymph node swelling, up to 1 cm in diameter.Compared with lupus mice called MRL/lpr,the MHX10/Mo-lpr/lpr mice have lighter glomerulonephritis, vasculitis, arthritis, salivary gland inflammation and longer life cycle.Based on these characteristics,MHX10/Mo-lpr/lpr mice are chosen to establish lymph node metastasis model; to inject chemotherapy drugs to the central of lymph node to kill residual cancer cells in the lymph vessels and lymph nodes though lymph drug delivery system;to establish diagnostic criteria of lymph node metastasis with small animals CT and MRI, and to provide reliable preclinical experimental animal model for early diagnosis of clinical LN metastasis.This article summaries MXH10/Mo-lpr/lpr mice genetic background, characteristic phenotype and application prospect.

The Progress of Conditional Knockout Animals and Its Application in Toxicology

2018, 35(02):  79. 

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In vivo animal experiments are crucial method and main content in the toxicology studies. Traditional knockout animals have certain limitations in the embryonic lethal genes researches. Conditional knockout animals overcome the defects existing in traditional knockout animal, which offers an important tool for the broader, deeper toxicology research. We will review two aspects including the principle of conditional knockout technology and its application in toxicology in this article.