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28 December 2021, Volume 38 Issue 6
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Veterinary Status of Laboratory Animal Industry
XU Hufeng, SUN Chenyang
2021, 38(6): 1.
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In the laboratory animal industry, veterinarians are involved in series of research works including the
daily management, protocol, animal health security and nursing, supervision and management of animal welfare,
animal experimental technology, sampling and euthanasia of laboratory animals’ operation, etc. Combined the
relevant literatures with own work, the current situation of laboratory animal veterinary is analyzed.
The Effect of Naringin on the Intestinal Flora of DSS-induced Colitis Mice
ZHANG Weidong, XU Yang, ZHENG Lu, et al
2021, 38(6): 5.
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Objective
To explore the effect of naringin ( NG) on the intestinal flora of mice with DSS-induced colitis.
Method
C57BL / 6 male mice were used to construct animal models. Fifteen mice were divided into the normal group, the model group and the NG group, each group with 5 mice. Except the normal group, the other 2 groups were given 4% ( m / v) DSS as freely drunk, and the NG group was given NG for intervention from the second day. The mice were sacrificed on the 9
th
day, and their colon and feces were collected. Western blot was used to detect the inflammatory molecule protein p-NF-κB-p65 and the apoptosis protein B lymphama 2 ( Bcl-2) . Using PCR-denaturing gradient gel electrophoresis ( PCR-DGGE ) technology to analyze the structure of mouse intestinal flora.
Result
The expressions of p-NF-κB-p65 protein and Bcl-2 protein in the model group were significantly different from those in the normal group and the NG group had different degrees of changes after NG intervention. The bands in PCR-DGGE were performed by unweighted pairwise arithmetic, the UPGMA algorithm was used for cluster analysis, and the result showed that the structure of the colonic flora of mice was significantly different in each group.
Conclusion
NG can effectively improve the symptoms of DSS-induced colitis and improve the intestinal flora of mice.
Effects of Oxygen Glucose Deprivation on Autophagy and Lipogenesis of Rat Adipose Stem Cells
YUAN Zhijian , WU Xiaoyu , HE Wenjuan , DING Yaming , ZHONG Xiaochun
2021, 38(6): 11.
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Objective
To study the autophagy of rat adipose derived stem cells ( ADSC) and effects of oxygen glucose deprivation on autophagy and lipogenesis of rat ADSC.
Method
The ADSC were prepared conventionally and set up as the control group ( group G) , the low glucose group ( group SG) , the hypoxia group ( group G + CoCl
2
) , the hypoxia & low glucose group ( group SG + CoCl
2
) and the autophagy positive control group ( group EBSS) . In different culture periods, cell viability was measured by MTT assays, the autophagy by Western blot and adipogenic induction, the oil red O staining and its OD value were performed.
Result
The ADSC in all groups were cultured and survived for 24 hours without difference (
P
>0. 05) in different culture periods. But at 48 and 72 hours, the cell proliferation of the group G was significantly higher than that of the other 3 groups(
P
<0. 05) . By Western blot, the protein expression of LC3-Ⅰ was significantly less than that of LC3-Ⅱ in groups G,SG,SG+CoCl
2
and EBSS. However, the LC3-Ⅰ s in groups G+CoCl
2
and SG+CoCl
2
were much higher than that in the group EBSS. In the test of adipogenesis, the OD value of the group G was the highest, which was statistically different (
P
< 0. 05 ) , compared with other groups.
Conclusion
The rat ADSC may have a high level of autophagy. It was suggested that under the circumstance of sugar deficiency and hypoxia, inhibiting autophagy will hinder the survival and lipogenesis of rat ADSC.
Preliminary Study on Anti-inflammatory Effect of Shegan Zhike Capsule
BAO Yulong, GAN Yu, FAN Yinglan, ZHU Jinghe, WU Xiaolin, LIU He, WU Yi, LI Guoxin
2021, 38(6): 16.
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Objective
To explore the anti-inflammatory effect and mechanism of Shegan Zhike capsule.
Method
The anti-inflammatory effect of Shegan Zhike capsule was observed by ear swelling test in mice and granuloma test in rats. The mechanism of anti-inflammatory action was speculated by detecting the levels of interleukin 6 ( IL-6) and tumor necrosis factor α ( TNF-α) in the serum of mice with lung injury induced by lipopolysaccharide ( LPS) .
Result
Compared with the blank control group, the low, medium and high doses of Shegan Zhike capsules could significantly reduce the degree of ear swelling in mice, and the difference was significant (
P
<0. 01 and
P
<0. 05) . Compared with the model control group, the low, medium and high doses of Shegan Zhike capsules could significantly reduce the weight of rat granulomas, and the difference was significant (
P
< 0. 01 and
P
< 0. 05) . Compared with the model control group, the low, medium and high doses of Shegan Zhike Capsules could significantly reduce the levels of serum IL-6 and TNF-α in mice, and the difference was significant (
P
<0. 01 and
P
<0. 05) .
Conclusion
Shegan Zhike capsule has certain anti-inflammatory effect and inhibiting inflammatory factors.
Effects of miR-143-3p in Thyroid Cancer by Regulating the Expression of TGIF2
LI Jia, LI Tao, PAN Jing, ZHAO Qian, LIU Li
2021, 38(6): 20.
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Objective
To explore the effect of overexpression of miR-143-3p on thyroid cancer ( TC) cells and its mechanism.
Method
The expression of miR-143-3p in normal thyroid epithelial cells and different TC cells was analyzed by Real-time PCR. Targetscan website and dual luciferase reporter system verified the targeting relationship between miR-143-3p and TGIF2 3′ UTR; the overexpression efficiency of miR-143-3p and TGIF2 mRNA was detected by Real-time PCR. After successful overexpression, CAL-62 cells were divided into the control group, miR-143-3p mimic group, pcDNA-TGIF2 group and miR-143-3p +TGIF2 group, the contents of inducible nitric oxide synthase ( iNOS) , interleukin ( IL) -1β and IL-6 in cell supernatant were detected by ELISA. The level of superoxide dismutase ( SOD) was detected by the kit, and the production of reactive oxygen species ( ROS) was detected by immunofluorescence. Micro-observation of stem cells into spheroids, and the expressions of TGIF2, p-P65, leucine-rich repeat-containing G protein-coupled receptor 5 ( LGR5 ) and octamer-binding transcription factor 4 ( OCT4) were detected by Western blot. The effect of miR-143-3p overexpression on tumor growth in nude mice was detected.
Result
The expression of miR-143-3p in TC cells was significantly lower than that in normal TC cells (
P
< 0. 05 ) . Targetscan website and dual luciferase reporter system confirmed that miR-143-3p was targeted to TGIF2. In the miR-143-3p mimic group, the expression of miR-143-3p was up-regulated (
P
<0. 05) , the expression of TGIF2 mRNA and protein was down-regulated (
P
<0. 05) , the contents of iNOS, IL-1β and IL-6 were significantly decreased (
P
<0. 05) , the antioxidant capacity and the sphere-forming ability of stem cells were significantly decreased (
P
<0. 05) , and the protein expression levels of p-P65, LGR5 and OCT4 were significantly decreased (
P
< 0. 05 ) . The results of pcDNA-TGIF2 group were opposite. Overexpression of miR-143-3p can significantly inhibit the pro-cancer effect of pcDNA-TGIF2. The xenograft tumor model in nude mice showed that overexpression of miR-143-3p could reduce tumor weight and volume, and down-regulate the expression of TGIF2 and OCT4 in tumor tissues (
P
< 0. 05) .
Conclusion
miR-143-3p overexpression can inhibit the levels of proinflammatory factors, antioxidant capacity and stem cell-like characteristics of TC cells by down-regulating the expression of TGIF2, and inhibit the formation of tumors in nude mice.
Protective Effect of Huangjia Ruangan Granules Against Immune Hepatic Fibrosis Induced by Human Serum Albumin in Rats
GAO Xiaoxin, ZHU Chunyue, ZHANG Qianqian, WANG Jiashuai, CAO Chunran, HU Yuchi, LIU Hongyan, LI Xuemei
2021, 38(6): 28.
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Objective
This study was designed to observe the protective effect of Huangjia Ruangan granules
against immune hepatic fibrosis induced by human serum albumin ( HSA) in rats.
Method
By vivo
experiment,
72 male rats were randomly divided into the normal group, the model group, the positive group, the high-dose
group, the moderate-does group and the low-dose group of Huangjia Ruangan granules,
n
= 12. The rat model of
immune hepatic fibrosis induced by HSA was established. The effects were evaluated by liver and spleen index,
liver fibrosis markers, serum biochemical index and liver histopathological examination.
Result
The content of
HA in 3 groups of Huangjia Ruangan was significantly lower than this in the model group, the levels of LN, PC Ⅲ ,
the contents of ALT and AST in the high-dose group were significantly lower than those in the model group. The
degrees of liver fibrosis in the high-dose group and the moderate-dose group were significantly improved comparing
with the model group.
Conclusion
Huangjia Ruangan granules can significantly improve hepatic fibrosis in rats
induced by HSA.
Comparison of Two Domestic and One Imported ELISA Kits for the Detection of Mouse Hepatitis Virus Antibodies
FANG Jiao, LIANG Jinning, FANG Xi, ZHU Kangli, HUANG Haina, LUO Qiulin, PAN Xuanxiao, GUO Xiaoping
2021, 38(6): 33.
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Objective
To compare the sensitivity, specificity, precision and reliability of 2 domestic and 1 imported ELISA kits for the detection of mouse hepatitis virus (MHV) antibody, so as to screen out a kind of economic and reliable ELISA kit.
Method
Three kinds of ELISA kits were chosen to detect MHV antibodies and other 5 kinds of viruses such as Sendai virus (SV), Peneumonia virus of mice (PVM), Minute virus of mice ( MVM), Poxvirus of mice ( MPV) and Reovirus type Ⅲ ( Reo-3) , and compared their sensitivity, specificity, precision and reliability.
Result
There were significant differences in the sensitivity of the 3 kinds of kits. Domestic kit B is 400 times as sensitivity as domestic kit A, and the imported kit is 4 times sensitivity of kit B. There were no cross reaction with SV, PVM, MVM, MPV and Reo-3 in domestic kit B and the imported one, but there were cross reactions with domestic kit A. The precision test showed that the average coefficient of variation in the imported kit was 5. 667%, and in the domestic kits A and B were 13. 825% and 13. 827%. The reliability test showed that the coincidence rates of positive and negative tests of domestic kit B and the imported one were both 100%, and the coincidence rates of positive and negative tests of domestic kit A were 40% and 80%, respectively.
Conclusion
The specficity and reliability of domestic kit B are consistent with the improted one, but with lower sensitivity and precison than it. Domestic kit A is lower in all tests than others.
Normal Reference Ranges of Blood Glucose Indexes among Pregnant SD Rats: A Systematic Review and Meta-analysis in Single Group Continuous Variables
JIANG Xinyong, GE Li, LAI Yuting, HUANG Pingping, CHEN Fan, PANG Shuqin, YU Honghong
2021, 38(6): 37.
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Objective
To identify the reference limitation of fasting blood glucose ( FBG) and fasting serum insulin ( FINS) among pregnant SD rats.
Method
Three Chinese databases and 6 English databases were retrieved from January 2014 to December 2020 including CNKI, Wan Fang, VIP, Web of Science, The Cochrane Library, Scopus, PubMed, EmBase and MEDLINE. The data in these obtained references were extracted based on the inclusion and exclusion criteria. The quality of these references was evaluated by SYRCLE’ s risk of bias tool for animal studies. The statistical software STATA 15. 0 was used to do the Meta-analysis of Single Group Continuous Variables. Weighted mean difference ( WMD) and 95% confidence interval ( CI) were used for combined statistic calculation.
Result
Four hundred and twenty-eight rats were included in these 37 articles. According to the systematic review and Meta-analysis, the upper reference limit of FBG and FINS on week 0, week 1, week 2 and week 3 among these pregnant rats were 5. 51 mmol / L( FBG) and no statistical data ( FINS) , 6. 92 mmol / L( FBG) and 15. 66 mIU / L( FINS) ,6. 64 mmol / L( FGB) and 15. 98 mIU / L( FINS) , together with 7. 04 mmol / L( FGB) and 15. 63 mIU / L( FINS) , respectively.
Conclusion
The reference limitation of FBG and FINS among pregnant SD rats determined by Meta-analysis would guide the related experiments about detecting FBG and FINS among pregnant SD rats, and thus reduce the error among experiments in the future.
Comparison of some Indicators among SD and Wistar Rats in Different Feeding Periods and Genders
TAI Gang, GUO Dong, YANG Zhimin, LI Yang, ZHANG Hong, LI Chen, WANG Tingting, LU Chao, ZHAO Sijun, ZHANG Junfeng, YANG Guowei
2021, 38(6): 47.
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Objective
In order to investigate the biological differences of some physiological and biochemical indexes between SD rats and Wistar rats in SPF level at different feeding periods and different genders, and to establish the normal reference ranges of blood physiological and biochemical indexes of different strains and ages.
Method
Three hundred SPF Wistar rats and 300 SD rats at the age of 5 to 6 weeks old were randomly assigned into 4,12 and 24 weeks groups by weight, with half male and half female. The blood physiological and biochemical indicators were detected by fully automatic biochemical analyzer and blood cell analyzer.
Result
The total WBC of female rats at 12-week group and male rats at 24-week group was significantly higher than 4-week group rats (
P
<0. 01) . The total WBC of male rats at 12-week group was higher than 4-week group rats (
P
<0. 05) , while the total WBC in 4-week and 12-week groups female rats was lower than that in male rats (
P
< 0. 05) . There was no significant difference in hematologic indexes of Wistar rats of different feeding period and genders (
P
>0. 05) . The serum ALP of SD rats at 4-week group was significantly higher than that of 12-week and 24-week groups (
P
<0. 01) , and the ALP of female rats at 4-week group was significantly lower than that of male rats (
P
<0. 05) , and the ALP in female rats at 12-week and 24-week group was significantly lower than that in male rats (
P
<0. 01) . TG in SD rats was higher than 4-week group (
P
< 0. 01) . The ALP in 12-week and 24-week groups female Wistar rats was significantly higher than 4-week group rats (
P
< 0. 01) , and 12 weeks male rats was significantly higher than 4 weeks rats (
P
< 0. 05) . ALP of female Wistar rats at 4-week and 24-week groups was significantly higher than that of male rats (
P
< 0. 01) , and ALP of female rats at 12-week group was significantly higher than that of male rats (
P
< 0. 05 ) .
Conclusion
Different feeding periods, genders and strains of SPF rats have some distinctions in blood physiological and biochemical indexes.
Study on the Application of Bullfrog Instead of Toad in Physiology Teaching Experiment
XU Caiyun, LI Tianshu, LI Xiaohui, YU Huimei, Guo Xinrong, ZHANG Dawei, CUI Jiarun
2021, 38(6): 53.
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Objective
This study explored the feasibility of using bullfrogs to replace toads in physiology teachingexperiments by comparing the experimental effects between bullfrogs and toads in physiology teaching experiments.
Method
Bullfrogs and toads were separately used to carry out frog heart perfusion experiment, sciatic nervegastrocnemius muscle stimulation and contraction experiment.
Result
The frog heart perfusion experiment showed that the bullfrogs and toads under the action of Na
+
, Ca
2+
, K
+
, norepinephrine, acetylcholine, lactic acid and sodium bicarbonate had the similiar effect on the activity of the isolated heart. In the sciatic nerve-gastrocnemius stimulating contraction experiment, the stimulation intensity was 0. 1 V caused minimal muscle contraction. Under the stimulation of around 0. 15 V, the muscle contraction amplitude reached maximum and remained urchanged. As the stimulation frequency increased, the skeletal muscles performed single contraction, incomplete tonic contraction and ionic contraction. After stimulated, the action potential conduction velocity of the nerve trunk about bulfrogs and toods is 16. 6 m / s and 13. 16 m / s respectively. There was no essential difference about experimetal results between the bullfrog group and the toad group (
P
>0. 05) .
Conclusion
It is feasible solution that bullfrogs instead of toads to carry out physiology teaching experiments.
Effect and Significance of Ventricular Structural Remodeling on Electrophysiological Function in Chronic Heart Failure Rabbits
AN Xiaoxia, HE Meinian, HE Xiuying
2021, 38(6): 59.
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Objective
To investigate effect and implication of cardiac performance and electrophysiological induced by structural remodeling with chronic heart failure( CHF) in ventricular of rabbits.
Method
Thirty two male New Zealand rabbits were randomly divided into 2 groups: the control group (
n
= 15) and the CHF group (
n
= 17) . The CHF group was used to prepare CHF model by injecting isoprenaline ( ISO) into rabbit ear vein, and finally 15 rabbits were successfully made. The control group was given the same volume of 0. 9% saline, and finally 12 rats entered the experiment. Echocardiography was used to monitor the M-type cardiac function of the 2 groups of rabbits, and lead Ⅱ ECG was recorded. LVEF, Avmax, LVEDd, LVESD, shortening rate of left ventricular short axis were recorded ( LVFS ) , interventricular septal thickness ( IVST ) , left ventricular end diastolic volume ( LVEDV) and left ventricular end systolic volume (LVESV), as well as heart rate, PR interval, QT interval and ST segment, were used to evaluate the changes of ECG physiological function. Programmed stimulation was used to observe the effective refractory period and dispersion of each effective refractory period when the stimulation perimeter (BCL) was 150 ms and 200 ms. To evaluate the BCL and the rate of ventricular arrhythmia (VA) induced by burst packing.
Result
Compared with the control group, LVEF, LVFS and Avmax of the CHF group were significantly lower, LVEDd and LVESD were significantly longer. IVST was thinner than that of the control group. LVEDV and LVESV were significantly higher than that of the control group (
P
< 0. 01) . HR of CHF group was significantly slower than that of the control group (
P
< 0. 01) . PR and QT intervals were significantly longer than that of the control group(
P
< 0. 01 ) . ST segment was significantly lower than that of the control group (
P
< 0. 01 ) . The difference was statistically significant. In the CHF group, ERP
150
and ERP
200
were significantly longer than those in the control group, and both of dERP
150
and dERP
200
were significantly larger than those in the control group (
P
<0. 01) . The BCL inducing VA was significantly longer than that in the control group, and the VA inducing rate was significantly higher than that in the control group, the difference was statistically significant (
P
< 0. 01 ) .
Conclusion
The ventricular structure and electrophysiological changes of chronic heart failure rabbits were significantly reconstructed, and the interaction between them increased the rate of reentrant VA induction.
Experience and Technique of Making Pathological Tissue Sections of Laboratory Animals
QIAO Xin, LI Meng, JIAO Kun, LI Shengli
2021, 38(6): 63.
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Objective
The quality of pathological sections of laboratory animals made in the early stage was analyzed to provide reference and guidance for improving the level of section making.
Method
A total of 1800 slices were randomly selected as the research objects, the failure rate was calculated according to the relevant quality control standards, and the causes of the failure were analyzed.
Result
The optimal slice rate was 90. 17% (1 623 / 1 800) , and the bad slice rate was 9. 83% ( 177 / 1 800) . The proportion of grade a film is 31. 50%, which should be further improved. The failure rate is within the acceptable range.
Conclusion
Standardized sampling operation, reasonable selection of fixed solution and fixed time according to the species and age of laboratory animals, strict control of embedding process, improvement of sectioning technology, timely replacement of reagent and other factors can help improve the quality of pathological sections of laboratory animals and facilitate scientific research.
Study and Establishment of Local Standard about Microbiology and Parasitology Levels and Monitoring in Laboratory Cattle
LI Changwen, ZHU Yuanmao, XIA Changyou, CHEN Hongyan
2021, 38(6): 66.
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Standardization is the basic property of laboratory animals. Quality standard is an important part of the quality control system, and the level of microbial and parasitic control of them is the fundamental basis for classification of laboratory animals. This paper introduces the background of the compilation of the local standards “ Laboratory Animal Cattle Levels and Monitoring of Microbiology “ and “ Laboratory Animal Cattle Levels and Monitoring of Parasitology” in Heilongjiang province which included in the setting of the level of laboratory cattle, the selection of testing items, the testing method and the expected social benefits. This standard can provide standard and guarantee for the new resources of agricultural laboratory animals in Heilongjiang province technically, several standerds, such as the important basic conditions for public health and biosafety, the high-quality laboratory animals for the research of major animal diseases, and the standardized laboratory animals for the research, production and verification of biological products. Then,it can promote Heilongjiang province and even the national animal husbandry industry technology progress and animal husbandry healthy development.
Stability Evaluation about Technical Parameters of EVC and IVC Systems
GENG Xiangfei, WANG Peng, LIU Pengjie, WANG Shuxin
2021, 38(6): 73.
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Objective
To compare the performance of closed exhaust ventilation central cages ( EVC ) and individual ventilated cage ( IVC) systems for rats and mice.
Method
Through the breeding space of the EVC system and ICV system, the operation process ( cage changing process and cleaning process) , damage rate, overall disinfection and cleaning process, mouse breeding environmental conditions ( number of air changes and wind speed and ammonia concentration ) , cage evaluation of indicators such as internal noise decibels, power outages or mechanical failures, and alarm systems.
Result
EVC cage covers a smaller area and easier to wipe and clean. The operation of observation, cage replacement and cleaning process are more simple than IVC system, escapically with lower accessory loss and energy consumption. But IVC cage performs better in washing, sound insulation,etc.
Study on Purification Technology of Genetically Engineered Mice Cesarean Section
ZHANG Man, PENG Lina, GAO Zefa, XU Wangjie, WANG Zhaoxia
2021, 38(6): 76.
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Objective
To obtain SPF healthy mice and improve the purification efficiency of cesarean section.
Method
The following experiments are involved in this study, such as judging the development of the fetus by palpation to accurately determine the start time of the cesarean section, by optimizing the cesarean section procedure, shortening the operation time to improve the efficiency of cesarean section, improving the milking method of the excised pup rate and doing statistical method to analyze the cesarean section purification data.
Result
A. By optimizing and improving the surgical procedure, the success rate of uterine section surgery reached ( 97. 9± 8. 25) %; B. when the time interval was less than 24 hours, the average lactation rate was 91. 2%; C. the microbiological grades of the pups purified by cesarean section technology reached the SPF level.
Conclusion
The perfect cesarean section technique not only improves the microbiological grade of laboratory mice, but also greatly improves the success rate of surgery, and laying the foundation for the biological purification of laboratory mice.
Application and Standardization of Laboratory Cats
CHEN Jun, WANG Liang, QIU Zewen, ZHAN Hongwei, LI Huiling, CHEN Dapeng, WANG Fujin, WANG Aiguo, WANG Jingyu
2021, 38(6): 80.
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As a medium-sized laboratory animal, cat has unique physiological and anatomical characteristics and is widely used in biology, medicine and other research fields. Cat can be used to prepare animal models of various diseases, and it has irreplaceable roles in the researches for nervous system and antihypertensive experiments. In recent years, although there are some progresses in the standardization of laboratory cat in China, it is relatively slow. In this paper, we review the application and standardization of laboratory cat in china based on the current situation of the research and application and the domestic and foreign progresses of standardization of laboratory cat.
