Table of Content

28 October 2019, Volume 36 Issue 06

Previous Issue    Next Issue

Influence of Naoshantong Capsule on Blood Lipid Levels in Atherosclerotic Mice

2019, 36(06):  1. 

Asbtract ( )   PDF (1268KB) ( )  

Related Articles | Metrics

To observe the effect of naoshuantong capsule on blood lipid levels in atherosclerotic mice and further explore its related mechanism of treatment on stroke. Methods Six weeks old ApoE-/- mice were fed with western diet for 12 weeks and randomly divided into three group: model group, naoshuantong low dose group and naoshuantong high dose group. The high and low dose group were given intragastric administration with 1.0 g/kg and 0.5 g/kg for 12 weeks, respectively. The model group were given normal saline of equal volume distilled water. After 12 weeks administration with naoshuantong capsule, serum was collected from the abdominal aorta for lipid levels measurement. The levels of serum lipids TC, TG, LDL-C and HDL-C were detected by enzyme method. Results Compared with model group, Naoshuantong capsule low group and high group both could decrease the levels of TC, TG, LDL-C significantly, but there was no influence on HDL-C. Conclusion Naoshuantong capsule could regulate the level of blood lipid, and its mechanism of treating stroke may be related to this pathway.

Establishment of nucleic acid detection method for experimental animal ferret Aleutian disease virus

2019, 36(06):  5. 

Asbtract ( )   PDF (1311KB) ( )  

Related Articles | Metrics

OBJECTIVE To establish a PCR detection method for Aleutian virus for the detection of ferrets Aleutian virus in experimental animals. Methods A pair of primers were designed with reference to the VP2 gene sequence of Aleutian disease virus in Genebank. The PCR method was established to test the specificity and sensitivity, and the experimental samples of Ferrets feces were screened. RESULTS The sequencing results showed that the designed primers can specifically amplify the 531 bp gene fragment of VP2 of Aleutian disease virus; There were no unspecific amplification when the following experimental animal DNA virus were used as templates in ADV PCR reaction: mouse Polyoma Virus, Minute virus of mice, Canine Parvovirus, Herpes Simplex virus, Ectromelia virus, Sendai virus, Mouse adenovirus; the nucleic acid plasmid with a sensitivity of 90.6 copy/μL was obtained. A total of 39 samples of ferrets were tested and no Aleutian virus nucleicacid was detected. Conclusion The current ADV PCR detection method shows applicable specificity and sensitivity, which can be used as a pathogen detection method in the screening of ferrets Aleutian disease virus in experimental animals.

Study on Inflammatory Response of the staphylococcus Aureus infected Deep Second Degree Scald Germ-free Rats

2019, 36(06):  9. 

Asbtract ( )   PDF (9614KB) ( )  

Related Articles | Metrics

We established a deep second-degree scald injury model which was infected with staphylococcus aureus with germ-free rats and SPF rats, then we analyze the healing process，inflammatory response，and pathological changes about the four groups. Deep second degree scald wound model was performed in 20 six weeks old female germ-free rats at a temperature of 94℃，8s in a clean bench . The scald germ-free rats were randomly divided into two groups. One group was infected with staphylococcus aureus with a concentration of 1×108CFU/mL in the wound；Another group was not infected.20 six weeks old female SPF Wistar rats were performed second degree scald wounds as the germ-free rats. The scab generating time， dislocation time and healing time of the four groups were observed，and serum samples were taken to detect the changes of IL-1α, TNF-α and EGFR before burned(0h) and 24h,48h,3d,5d,7d,10d,14d after burned. The skins were taken to observe the pathological changes at 24h,3d,7d and 10d after burned. The result showed that the staphylococcus aureus stained germ-free rats had a short period of decrustation and healing time (P＜0.05),while the staphylococcus aureus stained SPF rats had a long period of scab generating time, decrustation and healing time (P＜0.05).Because of the double damage, the level of TNF-α and IL-1α were elevated and they were significantly higher than the other three groups after 72h scald (P < 0.05);The germ-free rats response stimulus to scald and staphylococcus aureus, so the level of TNF-α、IL-1α and EGFR were elevated and they were significantly higher the SPF rats at 24h after burned. The level of EGFR about the staphylococcus aureus stained germ-free rats was significantly higher than SPF rats at 48h after burned. The pathological showed time of new blood vessels and granulation tissue formation about the staphylococcus aureus stained SPF rats were significantly longer than the other three groups and time of new blood vessels and granulation tissue formation about the staphylococcus aureus stained germ-free rats were significantly shorter than the other three groups. In conclusion, The immune system of germ-free rats was in a state of dormancy, when it was stimulated by scald and staphylococcus aureus, the systemic immune was activated and the wound healing was accelerated; The variety of immune evasion molecules secreted by staphylococcus aureus aggravated the inflammatory response and wound healing was delayed.

Effect of different mouse hepatitis viruses antibodies detected by ELISA

2019, 36(06):  15. 

Asbtract ( )   PDF (5652KB) ( )  

Related Articles | Metrics

Objective To establish the Mouse Hepatitis Virus (MHV) serum antibody ELISA test method for routine monitoring of MHV locally, in order to make timely judgments on the infection status of SPF mice. Methods Three kinds of MHV strains (MHV1, MHV-JHM and MHV-A594) were amplified in the L929 cells and purified by ultracentrifugation method. The appropriate virus antigen type was determined and an enzyme-linked immunosorbent assay (ELISA) system was established. SPF BALB/c mice were immunized to obtain high titer immunopositive serum. The purified MHV antigens were coated in microtiter plates and the detection system of ELISA flat was also optimized to apply clinical mice serum detection．Results The suitable coating antigen for local region was MHV1 and the methods for virus amplification and antigen purification were established. The prepared MHV antiserum keep high titer and can be used as standardized quality control serum. The optimal working concentrations of coated antigen, serum to be tested and enzyme conjugate were 4.0μg/mL (10-7.73/0.1mL TCID50), 1: 40 and 1:4000 respectively. The inter-assay and intra-assay average coefficient of variation is 5.13% and 5.57%, respectively. The detection sensitivity reach more than 1: 4000. There was no cross-reactivity with Sendai Virus (SV), Pneumonia Virus of Mice (PVM), Reovirus type III (Reo3), Minute Virus of Mice (MVM) and Ectromelia Virus (Ect). The relative deviation of stability test was less than 10%. A totally 165 sera samples were analyzed by the current ELISA method, the positive serum coincidence rate was 97.37% (37/38) and the negative serum coincidence rate was 92.19% (118/127), compared with the result of the commercial ELISA kit. Conclusion We have established ELISA method detection of mouse serum MHV, with high specificity, sensitivity and reproducibility It can be used for daily pathogen monitoring of MHV in this region, so as to make accurate judgment on the infection status of mice.

Model Depression for Chronic Mental Stress

2019, 36(06):  23. 

Asbtract ( )   PDF (1068KB) ( )  

Related Articles | Metrics

Objective Depression model was established with long-term mental stress stimulation. Methods 14 male C57BL/6J mice aged 4 weeks (body weight 11.05 1.65 g) were randomly divided into control group and LTMS group. In the LTMS group, the cats were kept in a single cage for 21 consecutive days and were given shock for 3h and sleep deprivation for 24h.The experimental group judged whether the modeling was successful or not by measuring the change of food intake, the overhead cross maze experiment, the open field experiment and the change of forced swimming behavior. Results Compared with the control group, the food intake of LTMS group was significantly reduced (p<0.01), and the forced swimming time was significantly prolonged (p<0.01).In the open-field experiment, the number of traversal and the number of upright position of LTMS group mice were significantly less than that of the control group, and the difference was extremely significant (p<0.01).In the overhead cross maze experiment, the residence time of the closed arm of LTMS group mice was significantly shortened, and the difference was extremely significant (p<0.01), and the proportion of the times of entering the open arm was reduced, and the difference was significant (p<0.05). Conclusions The long-term mental stress model can be used as the depression model because it shows the similar symptoms of behavior despair, decreased activity ability, loss of interest, anxiety and clinical manifestations of depression.

Determination of Growth curve and Hematological Parameters of SPF Expansion Stock Hartley Guinea pigs

2019, 36(06):  27. 

Asbtract ( )   PDF (1356KB) ( )  

Related Articles | Metrics

Measured the growth weight and blood physiological & blood biochemical parameters in SPF Hartley guinea pig core population. Compare with the hematological parameters of clean guinea pig. The results show that the WBC、NEUT、EOS have significant difference (P＜0.05), the PCT、MPV、LYM、LYM%、NEUT%、EOS%、BAS% have very significant difference(P＜0.01）between SPF male and female in blood physiological parameters. The ALP has significant difference（P＜0.05），the 3 indexes as BUN、CREA、CHO have very significant difference(P＜0.01）between SPF male and female in blood biochemical parameters. The results of comparison with CL guinea pigs show that WBC between SPF and CL of both sex, MCV、PLT between SPF and CL of female have no significant difference(P＞0.05），other indexes have very significant difference (P＜0.01）between SPF and CL guinea pigs. Conclusion, the growth curve of SPF guinea pig was plotted. The hematological parameters of SPF guinea pigs are significantly different from sex and have very significant difference between SPF and clean guinea pigs.

Establishment and Application of Fluorescent Quantitative PCR assay for Mouse Cytomegalovirus

2019, 36(06):  33. 

Asbtract ( )   PDF (5675KB) ( )  

Related Articles | Metrics

Objective To establishment and apply fluorescent quantitative PCR method for mouse cytomegalovirus. Methods Primer probes were designed by using the conserved sequence of MCMV Smith strain DNA polymerase gene published by NCBI to establish a quantitative PCR method for MCMV. The specificity, sensitivity, repeatability and stability of the method were verified. The sample was applied to detect mice samples mixed with MCMV and 409 mouse blood samples sent in 2018.Results The established FQ-PCR method for MCMV has a standard curve Slope of -3.418, an R2 value of 0.999, and an amplification efficiency of 96.137%. The minimum detectable amount of MCMV was 47 copies. There was no amplification curve when rat cytomegalovirus, monkey cytomegalovirus, human herpes simplex virus, pseudorabies virus and cat herpesvirus type I were used as templates, which showing the good specificity. The intra- and inter-assay coefficients of variation were 0.39%-0.68% and 0.48%-1.01%, respectively, with good repeatability and stability. The maximum dilution of MCMV virus in the blood samples of mice was detected to be 1:1000 (100.75 TCID50/0.1 ml), and 409 mouse blood samples were tested negative. Conclusion The established FQ-PCR method for mouse cytomegalovirus has good sensitivity, specificity and stability, and can effectively detect MCMV in mice, which provides technical reference for the monitoring of MCMV in lab mice and improvement of related standards.

Comparative study on pulmonary fibrosis model induced by bleomycin and endotoxin in two strains of mice

2019, 36(06):  39. 

Asbtract ( )   PDF (10046KB) ( )  

Related Articles | Metrics

A suitable mice pulmonary fibrosis model was established by comparing the degree of pulmonary fibrosis induced by bleomycin and endotoxin in two strains of ICR and C7BL/6 mice. Two strains of ICR and C57BL/6 mice were randomly divided into control group, bleomycin group and endotoxin group, respectively. The bleomycin group received a single intratracheal instillation of 5 mg/kg body weight of BLM to induce pulmonary fibrosis, the endotoxin group was Intravenously injected with endotoxin of 40 EU/kg, while normal control groups were administrated the same amount of normal saline. On the 7th, 14th and 28th day, 8 mice were respectively sacrificed in each model group. The general condition of the mice was observed, and the lung coefficient of the mice was measured. The pathological changes of the lung tissues were observed by HE staining. The content of hydroxyproline in the lung tissues was detected by ELISA. Compared to the control group, the lung coefficient and lung hydroxyproline content of each model group increased to different degrees (P0.05). C57BL/6 mice are more sensitive to bleomycin and endotoxin. Both bleomycin and endotoxin can prepare lung fibrosis models of acute lung injury in mice.

Effects of Astragalus Polysaccharides on Growth Performance, Blood Physiological and Biochemical Indexes and Incidence of Diarrhea in Breeding Rhesus Monkeys

2019, 36(06):  44. 

Asbtract ( )   PDF (1445KB) ( )  

Related Articles | Metrics

Objective To study the effect of Astragalus Polysaccharides on the growth performance, blood physiology, biochemistry and diarrhea incidence of breeding macaque. Method Sixty breeding macaques of different ages were fed with Astragalus Polysaccharides once a day. Before the experiment, 1 month after the experiment and 3 months after the experiment, the physiological and biochemical indexes, body weight and incidence of diarrhea were measured. Result With the increase of Astragalus polysaccharides , the body weight and the number of white blood cells and lymphocytes in blood routine were increased. In blood biochemistry , glutamic oxaloacetylase, glutamic pyruvic transaminase, total bilirubin, urea, total cholesterol, triglyceride, high- density lipoprotein cholesterol, low-density lipoprotein cholesterol, magnesium, C3 , C4, IgA and IgG all showed an upward trend , while blood glucose showed a down ward trend .With the increase of Astragalus polysaccharide , the number of diarrhea in breeding macaque also decreased. Conclusion Astragalus polysaccharide can obviously improve the feed utilization rate of breeding macaque, effectively regulate immunity, and also has a good prevention and treatment effect on diarrhea of breeding macaque.

The Normal Reference Value of Body Weight and the Rate of Body Weight Gain in Wistar Rats Aged from 5 to 109 Weeks

2019, 36(06):  50. 

Asbtract ( )   PDF (2150KB) ( )  

Related Articles | Metrics

Objective To learn about the body weight and rate of body weight gain of Wistar rat during most of its life. Methods The control group of a combined chronic toxicity and carcinogenicity test included 60 male and 60 female Specific Pathogen-Free Wistar rats (4 weeks old). The rats were acclimated for 1 week prior to initiation of the test. The rats were dieted with conventional feed. The study duration was 104 weeks. Body weight were measured weekly for the first 13 weeks and once every 4 weeks thereafter. Results The body weight and rate of body weight gain of Wistar rats at 37 time points during their life from 5 to 109 weeks were reported. (1) With the increase of age of the rats, the body weight increased continuously; males and females reached their maximum body weight at 93 and 102 weeks, which were 755.7±103.4g and 515.4±96.6g, respectively; their body weight decreased thereafter. (2) The rate of body weight gain decreased with the increase of rat age; the negative rates were first observed at 68 and 82 weeks for males and females, respectively; the negative rate became more negative finally. Conclusions The data about the normal reference body weight and the rate of body weight gain of Wistar rats at 37 time points during their life from 5 to 109 weeks reported in this paper may further improve and supplement the existing literature on Wistar rats′ normal reference body weight; and provide some useful reference for its inter-laboratory comparison and for the relevant technical staffs.

Application of improved cigarette smoke exposure method

2019, 36(06):  55. 

Asbtract ( )   PDF (1390KB) ( )  

Related Articles | Metrics

Objective: To investigate the effect of the modified method of smoke exposure on the survival rate of mice. Methods: on the basis of the common smoke exposure method, a device for increasing the oxygen quantity and increasing the oxygen amount at the bottom of the contaminated tank and monitoring the temperature simultaneously was made. The object of Kunming mice were divided into control group (not exposed in the common exposure device, exposure group (1), normal exposure method, exposure group (2) were improved) experiment, analyzing the application of spss20.0 statistical software were compared between the survival rate of mice, with oxygen concentration toxic unit and the changes of total the concentration of suspended particles. Results: a total of 160 Kunming mice were selected and randomly divided into 40 groups, 60 rats exposed to 1 groups, 2 rats exposed to 60 groups, both sexes were half and half. The experimental results showed that the survival rate in 2 groups of mice was significantly higher than that in group 1, the difference was statistically significant (x2=7.647 P=0.02<0.05), the oxygen concentration in the 1 groups were significantly lower than the exposure device of oxygen concentration of 2 groups, 2 groups of oxygen concentration fluctuations in the exposure and the device is small, the difference was statistically significant (F=5.260, p=0.045<0.05). There was no statistical difference in the concentration of TSP between the 1 groups and the 2 groups, indicating that the improved smoke exposure device did not affect the dose and exposure process of the toxicant. Conclusion: the improved smoke exposure device will reduce the mortality of the experimental animals under non pharmacological factors, and does not affect the experimental exposure process.

Research progress on evaluation methods of stress depression animal models

2019, 36(06):  59. 

Asbtract ( )   PDF (1375KB) ( )  

Related Articles | Metrics

Depression is a common mood disorders caused by a variety of reasons, with significant and lasting low mood as the main clinical features, and the low mood don’t match the situation. Some serious patients can appear suicidal thoughts and behaviors. The study of animal models of depression is the basis of the mechanism study of depression and the development of clinical drugs. Among them, stress model has become the most commonly used animal model of depression, which is widely used in basic research of depression, because the inducement that simulates depressive illness is quite sufficient and the modeling method is simple. In this paper, the main methods to evaluate stress animal models are summarized, which provides a reference for the modeling of stress depression animal models.

Nutrition characteristics and quality influencing factors of laboratory animals feed

2019, 36(06):  69. 

Asbtract ( )   PDF (1285KB) ( )  

Related Articles | Metrics

Laboratory animal feed is an important supporting condition closely related to the quality of laboratory animals. It is also a precondition to ensure the smooth progress of animal experiments and the accurate and reliable results. In this paper, the nutrition characteristics of laboratory animals feed are briefly described, and the main factors affecting the quality of laboratory animal feed are summarized.