Table of Content

28 February 2020, Volume 37 Issue 01

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Evaluation on the Genetic Quality of Tiger Skin Population by Microsatellites

MA Dong-Sheng, LI Hong-Yan, YAN Ke-Song, ZHAN Da-Wei

2020, 37(01):  1. 

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Objective To evaluate the genetic structure of Tiger skin population by microsatellites. Method 37 microsatellites were applied to amplify the 25 genomic DNA by PCR. Software POPGEN1. 32 was used to analyze the scanning result of PCR products. The parameters of genetic structure of Tiger skin population were obtained. Result The observed average number of alleles is 4. 702 7. The average effective average number of alleles is 2.6907. The average effective heterozygosity is 0.602 1. The average polymorphism information content ( PIC) is 0. 552 0. Conclusion Microsatellites can be used to genetic quality monitoring of Tiger skin population. The genetic background of Tigers kin population is in conformity with the characters of closed colony.

Establishment of Tetanus Disease Model in ICR Mice

2020, 37(01):  7. 

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Objective The aim of this paper is to establish the model of tetanus and complete the research of Clostridium tetani. Method ICR mice were used as the experimental animal model, and tetanospasmin ( TeNT ) was injected intraperitoneally. The behavior change and tissue (lung, liver and brain ) comparison of each group were observed in this experiment. Result Tetanospasmin can cause the body spasm , limbs rigidity spasmodic and opisthotonus , also cause liver to appear cell denaturation , cerebrum neuron necrosis , cause mice to die. However , when the dose of Tetanospasmin is lower than 10 4 test dose, it will not cause death in mice, and there is no obvious pathological change. Conclusion The model of tetanus disease in ICR mice was established.

The effect of HDCP on Type I lodothyronine Deiodinases and Thyroid Hormone

2020, 37(01):  11. 

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To investigate the effect of HDCP on DIO- I and thyroid hormone. Method Twenty pregnant rats were divided into control group ( n = 10 ) and model group ( n = 10 ) . On the 13 th day of gestation , rats in model group were received L-NAPE by subcutaneous injection until 21th day of gestation , HDCP model was reproduced. Control group received saline by the same ways and periods as model group. To get the livers ample and serum, the expression of DIO- I mRNA in liver and thyroid hormone level in serum were detected. Result Compared with control group, the expression of DIO- I mRNA in liver of model group was significantly decreased (P<0. 05 ) , and the TT3 , FT3 level in serum of model group were notably dropped ( P<0. 05 ) . Conclusion HDCP can result in the reduction of the expression of DIO-I mRNA in rat liver , and then the serum level of TT3 and FT3 are both dropped .

Determination and Analysis of Physiological and Biochemical Parameters of hVEGFR2-KI Mice

2020, 37(01):  15. 

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Objective To determine the blood parameters of hVEGFR2-KI mice at 4 weeks and 8 weeks of age of different genders, and to analyze the influence of gender and age on the blood parameters of this strain of mice. Method Venous blood was collected from 20 male and female mice aged 4 weeks and 8 weeks respectively. The physiological indexes of hVEGFR2-KI mice were determined by Japanese photoelectric Mek7 222 blood cell analyzer and Olympus AU400 automatic biochemical analyzer. Result Among the physiological and biochemical indexes measured, there was significant deference in hemoglobin concentration ( HGB ) , total protein ( TP) , total cholesterol ( CHO ) between male and female of 4-week-old hVEGFR2-KI mice ( P<0. 05 ) . There were significant differences( P<0. 01 ) in erythrocyte count ( RBC ) , hematocrit ( HCT ) , mean erythrocyte hemoglobin ( MCH ), mean erythrocyte hemoglob in concentration ( MCHC ) .There were significant differences in 11 physiological and biochemical indexes between male and female of 8 -week -old hVEGFR2-KI mice , which were blood physiological indexes including RBC , HCT ,HGB , platelet count , ( PLT ) , platelet volume , ( PCT ) , neutrophil absolute value (NEUT) , monocyte percentage ( MON%) and were 4 blood biochemical indexes such as CHO, alkaline phosphatase ( ALP) , glucose ( GLU) , TG; There are five statistically significant indicators, including percentage of lymphocytes ( LYM% ) , percentage of neutrophils ( NEUT% ) , TP , urea ( BUN ) and phosphorus( P ) . Differences of hVEGFR2-KI mice at different weeks of age were more widely distributed. They has a significant difference with RBC , HCT , MCH , HGB , white blood cell count ( WBC ) , average red blood cell volume ( MCV ) , red blood cell distribution width ( RDW) , mean platelet volume( MPV ) , platelet distribution width ( PDW) , lymphocyte absolute value ( LYM ) , monocyte percentage ( MON % ) , TP , CHO , albumin ( propagated ) , ALP , creatinine ( CREA) , GLU , P , a total of 18 indexes. PLT , LYM% , NEUT% , ALT and TG were the 5 indexes with statistical significance. Conclusion The physiological and biochemical indexes of hVEGFR2-KI mice aged 4 weeks and 8 weeks were determined in this study , which provided some reference for the application ofhVEGFR2-KImice and the standard of normal blood parameters.

Establishment of Experimental Animal Model of Chronic Pulmonary Inflammation Combined with Mycoplasma Pneumoniae Pneumonia

2020, 37(01):  22. 

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Objective Mycoplasma pneumoniae pneumonia ( MPP ) is an interstitial pneumonia caused by Mycoplasma pneumoniae ( MP ) with fibrous connective tissue hyperplasia in the pulmonary interstitial as the main pathological change , in clinically susceptible patients with chronic lung inflammation. For this reason , the study used a method of smoking and Mycoplasma pneumoniae infection to establish an experimental animal model of Mycoplasma pneumoniae interstitial pneumonia. Method Realtime PCR , HE and Masson staining were used to observe the concentration of Mycoplasma pneumoniae and the deposition of cellulose in lung tissue after infection in mice. Result In the smoked and infected group, the infection rate of Mycoplasma pneumoniae was 93%, the alveolar cavity collapsed , a flaky dense structure in the lung tissue , and thered blood cells filled with alarge amount of cellulose. Conclusion The lung tissue lesions of the smoked and infected mice are severe , showing a state of cellulose deposition .The model of chronic lung inflammation combined with Mycoplasma pneumoniae interstitial pneumonia was successfully established.

Protective Effect and Mechanism of PTS Preconditioning on Myocardial Ischemia-reperfusion Injury in Rats

2020, 37(01):  27. 

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Objective To investigate the protective mechanism of ginseng triolsaponin ( PTS) preconditioning on myocardial ischemia-reperfusion injury ( MIRI) in rats. Method After one week of adaptive feeding, 120Wister rats were randomly divided into a sham operation group ( normal saline 2 mL/kg) , a model group ( normal saline 2 mL/kg) , a PTS low dose group ( 25 mg/kg) , and a PTS medium dose group. (45 mg/kg) , PTS high dose group (90 mg/kg) , positive drug group ( shengmai injection 450 mg/kg) .The drug was administered intraperitoneally for 7 consecutive days. MIRI modeling was performed 1 hour after the last administration , and the materials were measured after 24 hours of reperfusion. Monitoring ST segment changes of rat electrocardiogram ； detecting serum creatine kinase isoenzyme ( CK-MB) , interleukin-6 (IL-6) and tumor necrosis factor-— ( TNF-—) levels; detecting myocardial tissue by PCR Bcl-2 and Bax mRNA expression levels ; myocardial histopathology and ultrastructural changes of cardiomyocytes were observed by light and electron microscopy. Result Compared with the model group , the PTS medium dose group and the high dose group can significantly reduce the STsegmentand serum CK- MB, IL-6, TNF-— levels in the MIRI model rats, improve the damage of myocardial structure, and increase the anti-apoptotic protein Bcl-2 expression inhibits the expression of the proapoptotic protein Bax. Conclusion PTS has protective effects on MIRI rat myocardium , and its mechanism maybe achieved by inhibiting the expression of inflammatory factors and regulating keyenzymes of apoptosis.

The Detection Strategy of Pathogenic Microorganisms in Experimental Animals─Mouse Hepatitis Virus

2020, 37(01):  33. 

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Objective To explore the study design of pathogenic microorganism infection in experimental animals , this study used two classical strategies to detect the change of virus load and serum specific antibody in faeces of MHE- infected mice. Method C57BL/6 and BALB/c mice ( n = 20 ) were divided into 4 cages and fed with litter from MHE- infected animals for 28 days. After which each animal was kept in an isolated cage with aseptic litter, the fecal and serum were collected per week. Samples were tested by Taqman real-time PCR and ELISA , and the data were analyzed statistically. Result For BALB/c mice, MHE was detected as early as d21 post infection (positive rate of 20%) and reached to the highest with 80% infection rate at d 28 , then decreased to 50% at d42, and can not be detected atd56 when using qPCR method .While positive result were detected atd42 with infection rate of 50% when using serum ELISA method. For C57BL/6 mice, MHE was detected at d28 and d42 post infection in only 20% of mice when using qPCR method , and no positive antibodies were detected during experimental period when using serum ELISA method. Conclusion For mouse hepatitis virus detection, BALB/c mice were recommended as sentinel mice in isolation quarantine and routine pathogen detection , and qPCR method was recommended for early isolation quarantinewith the advantage of earlier and higher detection rate.

Comparison of Disinfection Effects of Six Common Disinfectants on Different Bacteria

2020, 37(01):  38. 

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To investigate the antibacterial effect of six kinds of disinfectants on Salmonella and Escherichia coli, to determine the tolerance of bacterias to disinfectants, and to provide important evidence for the scientific use of disinfectants in the barrier environment of laboratory animals. Method Salmonella and Escherichia coli were isolated and the bacteriostatic and bactericidal effects of various disinfectants were determined by filter paper method . Result The result showed that 0. 05% Goudian had significantly higher bacteriostatic effect on Salmonella and Escherichiacoli than other disinfectants (P<0.05 ) , followed by 0. 05% Baidu and 0. 25% glutaraldehyde solution , 0.2% peracetic acid , 0.1% sodium dichloroisocyanurate powder and 0.25% neogel. Conclusion In this experiment , 0.05 % Goudian is the most effective disinfectant for Salmonella and Escherichia coli , which can provide important reference for the future use of animal barrier environment disinfection .

Effects of Glucocorticoid on Pulmonary Function Damage, Inflammatory Response and JAK2/STAT5 Pathway in Neonatal Mice Induced by Hyperoxia

2020, 37(01):  42. 

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Objective To explore the effects of glucocorticoid ( Glue) on pulmonary function injury, inflammatory response and JAK2/STAT5 pathway in neonatal mice induced by hyperoxia, and to provide theoretical basis for the treatment of pulmonary function injury. Method The mice were randomly divided into control group, hyperoxia group, low dose Gluc group, medium dose Gluc group and high dose Gluc group. The ventilation volume, airway resistance , airway pressure , lung volume and maximal inspiratory flow were measured by animal pulmonary function analysis system. HE staining and TUNEL staining were used to observe the lung tissue morphology and cell apoptosis in mice; RT-PCR, Western Blot were used to detect the expression of α-SMA and TGF-β； Elisa was used to detect the contents of IL-6, MCP-1 and iNOS; Western Blot was used to detect the phosphorylation of JAK2 and STAT5 .Result Compared with the control group , there sting ventilation volume , airway resistance , airway pressure , lung volume and maximal inspiratory flow of hyperoxia group mice were significantly decreased (P<0. 05) . Apoptotic number, expression ofα-SMA and TGF-β, EOS ratio, IL-6, MCP-1 and iNOS contents were significantly increased ( P <0.05 ) , and JAK2, STAT5 phosphorylation levels were significantly increased (P<0. 05) . However, medium and high doses of Gluc could significantly reverse the effects of hyperoxia on the above indexes in mice. In addition, hyperoxia group mice showed structural disorders of pulmonary parenchyma , enlarged alveolar septum and other morphological changes. In low , medium and high dose Glucuc group , the degree of tissue morphological changes gradually decreased. Conclusion Glucocorticoids can also alleviate lung tissue injury and down-regulate the content of inflammatory factors in lung tissue of model mice, and inhibit JAK2/STAT5 pathway activity , which has a good therapeutic effecton hyperoxia-induced lung tissue injury.

Exploring the Relationship between Dkk-1 Level and β-catenin in Serum of Postmenopausal Osteoporosis Rats

2020, 37(01):  50. 

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Objective To explore the level of β-catenin in serum of postmenopausal osteoporosis rats and analyze its relationship with Dickkopf1 protein. Method Sixty-two postmenopausal osteoporosis rats were selected as the observation group , and 46 postmenopausal healthy rats were selected as the control group , and relevant analysis was performed .The relationship between β-catenin and related parameters was analyzed. Baseline data of the two groups of rats were compared . Logistic regression analysis was performed on the factors that may affect osteoporosis in postmenopausal rats. Result There was no significant correlation between serum β-catenin and nuclear factor kB receptor activating factor ligand levels in the two groups ( P>0. 05 ) . There was no significant correlation between serum β-catenin and Dickkopf1 protein levels in the control group ( P >0. 05 ) . Serum β-catenin was negatively correlated with Dickkopf1 protein in postmenopausal osteoporosis rats ( P 0. 05 ) . The β-catenin ( 23. 23 ±0. 07 ) pg/mL of the observation group was significantly lower than that of the control group, and the data were statistically significant (P<0. 05 ) . The nuclear factor kB receptor activating factor ligand ( 138. 24±96. 45 ) pmol/L and Dickkopf1 protein ( 182. 79±34. 81 ) pg/mL were significantly higher in the observation group than in the control group. The data were statistically significant (P<0.05 ) . High-level nuclear factor kB receptor activating factor ligand is an independent risk factor for osteoporosis in post menopausalrats and Dickkopf1 protein, and β-catenin is a protective factor for osteoporosis in postmenopausal rats ( P<0.05 ). Conclusion β-catenin and nuclear factor kB receptor activating factor ligands play an important role in the pathogenesis of osteoporosis. The levels of β-catenin and Dickkopf1 are related to the process of osteoporosis in postmenopausal rats , while the level of β-catenin is related to Dickkopf1 protein is positively correlated .

Effects of PM2.5 on Airway Morphology and DNA Promoter Methylation of FOXP3 Gene in Asthmatic Rats

2020, 37(01):  54. 

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Objective To investigate the effects of PM2. 5 exposure on airway morphology and DNA promoter methylation of FOXP3 gene in asthmatic rats. Method 50 SD rats, 10 control rats and 40 asthmatic rats were selected. According to the different doses of PM2C5 , they were further divided into simple asthma group , low-dose group , medium-dose group and high-dose group .The effects of different doses of PM2.5 on the course of asthmain rats were compared. Result 1 ) Compared with the control group and asthma group , the weight of rats in all intervention groups was significantly lower ( P0. 05 ) . Compared with the control group and asthma group , with theincreaseofPM2.5 intervention dose , theeosinophil , neutrophil and lymphocyte counts in BALF of rats also showed an upward trend , while macrophages showed a downward trend (P<0.05 ) . 3 ) With the increase of the intervention dose, the methylation level and the relative expression of mRNA in the five groups decreased gradually. Conclusion PM2. 5 can decrease the methylation level of DNA promoter region and the relative expression of FOXP3 gene.

A New Method for Collecting Blood from the Jugular Vein of Rats

2020, 37(01):  59. 

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In the case of no anesthesia, rats were placed in a supine position to fix their trunk parts. Then the head was fixed with a plastic bottle, and the head was fixed. The self-made fixed head mask can effectively fix the rat, and establish a new method of collecting blood for the jugular vein of rats, so that the blood collection of the jugular vein of the rat is smooth. This new fixed method of fixing the rat by fixing the head cover can effectively prevent the operator from being bitten by the rat and ensure the repeated blood collection.

Common Problems and Countermeasures of C57BL/6 Mice in the Feeding Process of Diet-Induced Obesity Model

2020, 37(01):  61. 

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Objective To find out the problems in the process of establishing obesity model of C57BL/6 mice , and find effective ways to solve these problems. Method The mice with ear infection , hair removal and trauma caused by fighting were given corresponding precautions or partial treatments to make them grow healthily. Result Mice that received prophylactic disinfection during ear marking had a reduced risk of ear infection , and as mall number of mice with ear infection were restored to health after disinfection .After isolation and disinfection , the fur of depilated mice restored its integrity , and mice with mild trauma restored to health. Conclusion The problems and counter measures mentioned in this paper can provide references for other relevant researchers in the process of establishing obesity model in C57BL/6 mice.

Gender-specific Differences on Basic Anesthetic Effect in Miniature Pigs

2020, 37(01):  66. 

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The purpose of this paper is to explore the effect of sex on basic anesthesia in miniature pigs. This experiment covered 30 Bama pigs and 24 Nongda II mini-pigs, half male and half female. All were injected intramuscularly in the neck with a dose of 200 mg Lumianning and 10 mg midazolam. Completed the basic anesthesia , observed and record the time taken for the animal to achieve the effect and the dosage of the additional drug. Finally in the Bama pigs group, the female needed an average initial dose of 30% ～50% compared with the male. And Nongda II mini-pigs needed 50%～100%. So the conclusion is gender-specific differences affects the amount of basic anestheticused and the effective time in miniature pigs. Increasing the dose of anesthetic to female individuals can shorten the period of anaesthesia and the time of operation.

Research Progress on the Therapeutic Effect of Traditional Chinese Medicine on APP/PS1 Transgenic Mice with Alzheimer’ s Disease

2020, 37(01):  69. 

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Alzheimer's disease, is a central nervous system degenerative disease, the senile plaques and neurofibrillary tangles are its main characteristics. Patients with cognitive impairment, they can not take care of themselves. AD is a serious threat to the elder's health and longevity, the disease prevention and treatment has become a hot spot of research at home and abroad. In the field of experimental research on the prevention and treatment of AD, the transgenic mouse model of APP/PS1 is widely used, because it can better reflect the pathological features and clinical manifestations of patients with alzheimer's disease, and is a relatively ideal AD animal model. This paper reviews the past five years of traditional Chinese medicine effective component, Chinese medicine compound of APP ; PS1 transgenic mice AD advances in there search of the rapeutic effects oasto provide reference for subsequent research of prevention and treatment of AD.

Standardization and Application Progress of Tree Shrew

2020, 37(01):  74. 

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Tree shrew is a new type of laboratory animal being developed in China, which has been widely used in life science research. It has a close relationship with human beings and many biological characteristics are similar to human beings. Establishing artificial population of tree shrews and carrying outstandardization research are important guarantees to make tree shrew laboratory animal. The standardization and application of tree shrews were briefly summarized in this paper.