Table of Content

06 August 2021, Volume 38 Issue 3

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To Establish a Dynamic Model of Hepatic Fibrosis Induced by Diethylnitrosamine and Explore its Pathological Mechanism in Mice

WANG Mingliang, PENG Yunru

2021, 38(3):  1. 

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Abstract: Objective A dynamic mouse model of liver fibrosis was established induced by diethylnitrosamine ( DEN) to investigate the pathogenesis and pathologic etiology of liver fibrosis and to explore the possibility of predicting the stage of liver fibrosis by dynamically monitoring serum biochemical indexes. Method Male C57BL / 6 mice, were intraperitoneally injected with DEN twice a week at a gradient dose for 8 weeks. During the whole experiment, serum and liver tissues of mice were collected on the 2nd, 4th , 6th and 8th weeks respectively, and the progression of progressive liver fibrosis in mice was monitored and evaluated by detecting the corresponding serum biochemical indicators, the content of hydroxyproline in liver tissues, and the preparation observation. To explore the pathogenesis of hepatic fibrosis, western blotting was used to detect proteins involved in the TLR4 / MyD88 / NF-κB signaling pathway, and proteins associated with fibrosis including TGF-β1 and α-SMA. Result Compared with the control group, the serum biochemical indexes ( AST, ALT, ALP, T-BIL, A / G, etc. ) related to liver functions in DEN treated mice showed significant difference from the 4th week, and the body weight decreased after treated with DEN significantly, liver index was significantly decreased and hydroxyproline content in the liver tissues remained at a higher level from the 4th week ( P<0. 01) . The histopathological result indicated that DEN treatment induced hepatic fibrosis in mice on the 4th week. Western blotting analysis showed that the liver expressions of TGF- β1 and α-SMA in DEN treated mice significantly increased from the 6th week, compared with the control group, TLR4 / MyD88 / NF-κB signaling pathway was activated during the process of fibrosis, as shown by increased TLR4, MyD88, TRAF6 and p65 NF-κB in nuclear, accompanying with decreased p65 NF-κB in cytoplasm. Conclusion The result suggests that the mechanism of hepatic fibrosis may be related to the activation of TLR4 / MyD88 / NF-κB signaling pathway, which further promotes the secretion of TGF-β1 and activates hepatic stellate cells resulting in a large amount of extracellular matrix deposition. This study suggests that the pathogenesis of liver fibrosis in mice induced by DEN may be related to inflammation and indicates that to preliminarily judge the stage of liver fibrosis through multiple serum biochemical indexes.




Establishment of CD40 Immune Checkpoint Humanized Mice and Their Application in Antibody Drug Research #br# #br#

ZHANG Jing, ZHANG Zan, ZHAO Lei, GUO Yanan, HUANG Rui, XU Lida

2021, 38(3):  10. 

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Abstract: Objective Laboratory animal model is an important tool in the field of biological drug development, including oncology research, antibody drug development and preclinical efficacy and safety evaluation. The aim of this study is to prepare CD40 humanized mice ( B-hCD40 ) and provide more possibilities for preclinical pharmacodynamics study and safety evaluation of CD40 antibody. Method B-hCD40 mice was constructed by CRISPER / Cas9 system. Then the expression of human CD40 was detected by RT-PCR and flow cytometry. A tumor model was well established basing on B-hCD40. Meanwhile, anti-tumor efficacy and safety of CD40 activated monoclonal antibodies were evaluated. Result Human CD40 was only expressed in B-hCD40, and the ratio of different lymphocytes had no significant differences compared to wild-type mice. CD40 monoclonal antibody could significantly inhibit tumor growth. The weight loss of mice of the treatment group may be caused by the liver and kidney toxicity of the antibody. Conclusion In our study, B-hCD40 were successfully prepared, which is an effective animal model for evaluating the efficacy and safety of CD40 antibody drugs.




Study on Acute Toxicity of Deinococcus radiodurans 

XU Qin, SONG Laiyang, ZHANG Lei, LI Jiajia , MA Na, JIANG Junfeng , LIU Aizhong, PU Fengping, DENF Pei, LIU Jiangwei, WANG Wei

2021, 38(3):  17. 

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Abstract: Objective To investigate the acute toxicity of Deinococcus radiodurans ( DR) and its broken products. Method The KM mice were administrated with 0. 4 mL / 10 g of 1×10⁹ / mL DR ( high concentration DR group) , 1×10⁷ / mL DR ( low concentration DR group) , 1 × 10⁹ / mL DR broken products ( high concentration DR debris group) , 1×10⁷ / mL DR broken products ( low concentration DR debris group) and Tryptone Glucose Yeast agar medium ( TGY medium group) , respectively. After intragastrically administrated twice a day every 8 hours, the mice were observed for 14 days, then the organ coefficient, body weight, blood routine ( white blood cell, lymphocyte, red blood cell, hemoglobin, hematocrit, mean corpuscular volume, platelet) and plasma biochemical indexes ( alanine aminotransferase, aspartate aminotransferase, total protein, creatinine, creatine kinase, magnesium, phosphorus, creatine kinase MB isoenzyme, lactate dehydrogenase) were used to analyze and evaluate the toxicity of DR. Result No abnormal secretion, defecation, respiration and movement were found in mice in different dose groups. From 0. 5 hour after administration to the end of the experiment, no abnormalities such as slow reaction, pupil change, eyeball protrusion, eyelid ptosis and skin color change were observed in the test mice.There were no significant differences in body weight, blood routine test and blood biochemical indexes in the trialgroups compared with TGY group. The spleen coefficient was significantly increased in the low-dose group, and the spleen pathological examination showed that there was no substantial lesion. Conclusion The live and broken DR have no acute toxicity to mice. This experiment provides experimental basis for clinical application of DR.




Comparison of Biological Characteristics of CB6F1-Tg(HRAS)Nifdc Mice and CByB6F1-Tg(HRAS)2Jic Mice

LIU Susu, WEI Jie, WANG Chenfei, YUE Bingfei, FAN Changfa

2021, 38(3):  23. 

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Abstract: Objective To determine the difference of organs weight and blood physiological of CB6F1-Tg( HRAS) Nifdc Mice and CByB6F1-Tg( HRAS) 2Jic Mice. Method For this experiment, we selected 8-week-old CB6F1- NIFDC and CB6F1-CIEA mice ( 10 females and males ) of each sex, weighed body mass and subsequently dissected and assayed their major organ masses and blood physiology. Result There are some differents between the CB6F1-NIFDC and CB6F1-CIEA models in some parts of the organs and blood physiological parameters.Female mice showed some differences in heart, liver, lung and kidney, male mice showed differences in liver, lung, kidney, thymus and left and right testicular index mass. Among female mice, there were significant differences in 7 indicators ( RBC, HGB, etc. ) in 22 blood physiological parameters ( P < 0. 01) ; among male mice, there were significant differences in 5 indicators ( RBC, HCT, etc. ) in 22 blood physiological parameters ( P<0. 01) . Conclusion In this paper, the organ indicators and physiological indexes of the two models were measured, which may provide a reference for the application of the self-built model in the safety evaluation experiments of new drugs.



Effect of Astragaloside IV on MDA, SOD and GSH-Px Levels in Lens Tissue of SD Rats with Diabetic Cataract 

XIE Yanyan, SUN Yabin

2021, 38(3):  29. 

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Abstract: Objective To explore the protective effect of astragaloside IV on oxidative stress injury of lens tissue in diabetic cataract rats. Method Thirty SD rats were randomly selected from 120 rats as the control group, and the remaining 90 rats were intraperitoneally injected with 55 mg / kg streptozotocin ( STZ ) solution to establish the diabetes models. After successful modeling, it was randomly divided into the diabetic model group, the low-dose intervention group and the high-dose intervention group. Each group was 30 rats. The low-dose and high-dose groups were given 120 mg / kg and 240 mg / kg of astragaloside IV for 12 weeks, respectively. The lens transparency was measured in the 3 groups of rats at the 18th and 24th weeks of the experiment. After the end of the 24th week, the lens tissues of the 4 groups of rats were taken to detect the levels of MDA, SOD and GSH-Px in the lens tissues. Result At the 18th and 24th weeks of the experiment, the lens transparency of the 4 groups of rats was found to be significantly increased in the diabetic model group compared with the control group ( P < 0. 05) . The lens opacity of the 2 groups was significantly lower than that of the diabetic model group. Rats in the model group ( P<0. 05) , the degree of lens opacity in the high-dose intervention group was significantly lower than that in the low-dose intervention group ( P<0. 05) . At the end of the 24th week, the contents of MDA, SOD and GSH-Px in the lens tissues of the 4 groups were detected. The content of MDA in the lens of the model group with cataract formation was significantly higher than that in the control group ( P < 0. 05) , the contents of SOD and GSH-Px in the lens was significantly lower than that in the control group ( P<0. 05) . The content of MDA in the lens of the 2 groups was significantly lower than that in the model group ( P<0. 05) , and the contents of SOD and GSH-Px were significant. The increase of MDA content in the low-dose intervention group was significantly lower ( P<0. 05) , and the SOD and GSH-Px levels were significantly increased ( P < 0. 05) . Conclusion Astragaloside IV can enhance SOD and GSH-Px activities in lens tissue of diabetic cataract rats, clear oxygen free radicals in lens tissue, inhibit lipid peroxidation and reduce MDA levels. It also has a certain effect on preventing lens opacity in diabetic cataract rats. The higher dose of astragaloside IV is more effective in the lower dose of astragaloside.




Effect of Hydromorphine on ERK1 / 2 Signal Pathway in Spinal Cord of Rats with Inflammatory Pain 

SHI Xiaoling, JIA Wei

2021, 38(3):  34. 

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Abstract: Objective To investigate the effect of hydromorphine on ERK1 / 2 signal pathway in spinal cord of rats with inflammatory pain. Method Thirty SPF male rats were randomly divided into three groups: the normal group, the model group and the hydromorphine group. Detection of thermal pain threshold and mechanical pain threshold were detected in rats of each group. TNF-α, IL-6 and IL-10 in serum were tested by ELISA. Western blot detected the ERK1 / 2 and p-ERK1 / 2 protein expression in spinal cord of rats in each group. The expression of ERK1 / 2 mRNA in spinal cord of rats in each group was detected by qRT-PCR method . Result Compared with the normal group, the content of serum inflammatory cytokines TNF-α and IL-6 in the model group was significantly higher than that in the normal group, while the content of IL-10 in the model group was significantly lower than that

in the normal group, and the thermal pain threshold was significantly shortened and the mechanical pain domain was significantly decreased in the model group ( P<0. 05) . The expression levels of ERK1 / 2 mRNA, ERK1 / 2 and p-ERK1 / 2 protein in spinal cord of the model group were significantly higher than those of the normal group ( P< 0. 05) . After injection of hydromorphine, compared with the model group, the contents of serum TNF-α and IL-6 were significantly decreased, while the content of IL-10 was significantly increased, the thermal pain threshold was significantly prolonged and the mechanical pain domain was significantly increased ( P<0. 05) . The expression of ERK1 / 2 mRNA, ERK1 / 2 and p-ERK1 / 2 protein decreased significantly in hydromorphine group ( P < 0. 05 ) .Conclusion Hydromorphine can effectively improve complete freund ’ s adjuvant-induced inflammatory pain in rats. Its anti-inflammatory and analgesic mechanism is related to its reduction of the production of inflammatory factors and inhibition of the activation of ERK1 / 2 signal pathway in rats.




Re-evaluation of Literature on Rat Liver Fibrosis Animal Model Induced by Carbon Tetrachloride 

LI Gaohui, LV Wenliang , YAN Tun, LI Juanmei, WU Qingjuan, ZHAO Ting

2021, 38(3):  39. 

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Abstract:To compare the current method of liver fibrosis in rats induced by tetrachloromethane, and to establish the best method of liver fibrosis in order to provide reference for scientific research. Method Cnki, Wanfang data and cqvip were used to search and obtain the related literatures, data and the experimental pathological results and serological examination were selected, the literature was graded according to pathology and Alanine transaminase

and aspartate transaminase rates. The relevant experimental factors are evaluated, and then the criteria of model induction are established. Result In the model group, the normal hepatic lobule structure was destroyed and the interlobular fibrous septum was formed. The serum ALT and AST indexes were significantly increased.Conclusion The reliable experimental scheme of hepatic fibrosis model induced by tetrachloromethane oil is reliable. species, sex, pathway of drug action and concentration of CCl4 all influence the experimental result.




Effects of Different Interventions on Cognitive Ability in ASD Rats and Possible Mechanisms 

WEI Meiqi , LIU Zhongmin, FENG Guangzhi

2021, 38(3):  47. 

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Abstract: Objective Observe the effects of aerobic exercise, enriched environment and their combination on cognitive function and expression of NRXN1, NL3 and Shank3 in hippocampus of rats with ASD, to explore the effect and possible mechanism of it on correcting autistic abnormal behavior. Method ASD RAT models were established by intraperitoneal injection of VPA into pregnant Wistar rats. Forty offspring rats were randomly divided into aerobic exercise group, enriched environment group, exercise + environment group and ASD model group with 10 rats in each group. A total of 10 rats were randomly selected as the control group. The Rats in the aerobic exercise group received swimming exercise intervention lasting 8 weeks, 6 times per week, each time in the same time period, lasting 90 minutes, and the rats in the enriched environment group were kept in an enriched environment cage with various toys for playing. The exercise + environment group received exercise intervention and was raised in enriched environment, while the ASD model group and the control group did not. The behavioral changes were observed by open field test and Morris water maze test, and the expressions of NRXN1, NL3 and Shank3 protein were detected by western blot the mRNA expression levels of NRXN1, NL3 and Shank3 in rat hippocampus were detected by RT-PCR. Result Compared with the control group, ASD model group, aerobic exercise group, enriched environment group and aerobic + enriched environment group had fewer perforation times ( P<0. 05) , longer escape latency ( P<0. 05) , shorter activity time of the original platform quadrant ( P<0. 05) , lower mRNA expression of NRXN1 and Shank3 ( P < 0. 05) . Compared with the ASD model group, the crossing

times of aerobic exercise group, enriched environment group and exercise + environment environment group were significantly increased ( all P<0. 05) , the escape latency was significantly shortened ( P<0. 05) , the activity time of the original plateau quadrant was increased ( P<0. 05) , and the mRNA expression of NR3, NRXN1 and Shank3 were significantly increased ( P < 0. 05, P < 0. 01) . There was no significant difference between aerobic exercise group, enriched environment group and exercise+environment environment group in crossing times, escape latency and activity time of plateau quadrant and NL3, NRXN1 and Shank3 mRNA expression ( P>0. 05) . Conclusion Aerobic exercise, enriched environment and the combination of both can improve the cognitive ability and correct abnormal behavior of ASD rats induced by VPA, the intervention effect of aerobic exercise combined with rich environment is better than pure aerobic exercise and pure aerobic exercise than pure rich environment. Its mechanism of action may be related to the NRXN-NLGN-SHANK pathway.




Development and Application of a Rapid Calculator for Laboratory Animal Facilities Parameters Based on VB

LIU Wei, ZHAO Minghai, HOU Fengtian, ZHANG Xiao, LIU Zhiguo, ZHANG Xinyan, LIANG Chunnan

2021, 38(3):  54. 

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Abstract: Laboratory animal facility environmental monitoring is a part of the facilities acceptance and implementation of administrative licensing system in experimental animals, but also the facility to monitor the use of units to understand facilities are important means. As the provider of environmental testing services for laboratory animal facilities, National Institutes for Food and Drug Control ( NIFDC) , after obtaining a large amount of on-site inspection data, it is necessary to complete the calculation and report production as soon as possible to achieve the commitment to quality and efficiency of customers. This paper introduces a main function and programming logic diagram of the laboratory animal facility environmental parameter quick calculator designed by the NIFDC facility environmental detecting team based on VB programming.




Reference Range of Four Tests of Coagulation in the Plasma of SD rats 

LIU Zhenqing, ZHAO Yuanyuan, REN Yiran, ZHANG Meiling, LIU Ying

2021, 38(3):  59. 

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Abstract: Objective To establish the reference range of four tests of coagulation in the plasma of SD rats in the medical laboratory of Beijing institute of Chinese medicine, Beijing university of Chinese medicine. Method Prothrombin time ( PT) , activated partial thrombin time ( APTT) , thrombin time ( TT) and fibrinogen ( FIB ) levels in the plasma of forty SD rats were measured using automated coagulation analyzer. Result The reference range of PT, APTT, TT and FIB was 14. 96 to 15. 86 s, 15. 25 to 18. 83 s, 43. 06 to 53. 18 s and 1. 94 to 2. 21 g /L with stable result, respectively. Conclusion The reference range of four parameters of coagulation in normal SD rats was established, which provides reference value for pharmacological experiments and is of great significance for the safety toxicological evaluation of SD rats.




Detailed Process and Experiences of Cerebral Ischemia-reperfusion Model in Rats 

WU Lin, WU Fang, LIU Yanan, ZHANG Na, TAO Chengbin, ZHANG Yanying, XU Xiaoyan

2021, 38(3):  62. 

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Abstract:At present to make rat middle cerebral artery ischemia and reperfusion experimental animal model of diversity, improved intraluminal suture method selection in the process of experiment is more, this method has been proved to have a surgery, trauma small relative to the characteristics of simple operation, even so, the method to build module and the success rate is still low, therefore, a paper describing improved intraluminal suture method matters needing attention and the details of the article is necessary, for the future in the rat brain ischemia reperfusion mode of experiment personnel guidance of experience and also provide reference for more people to communicate. As a result , we explored and summarized the process and details of the experiment, and pointed out the details that need to be paid attention to in the experiment process, so as to provide a shortcut for the

experimenters who will make this model.




Anatomical Characteristics of Pancreas in Bama Minipigs 

ZHAO Yuqiong, DAI Xin, CHEN Hua

2021, 38(3):  66. 

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Abstract:The pancreas is an important digestive organ. Previous anatomical and histological studies on porcine pancreas mainly focused on the distribution of blood vessels and ducts. In this research, 24 control animals in another experiment were used to observe and measure the histo-morphological data of the pancreas of Bama minipigs. The result showed that the pancreas morphology of Bama minipigs is quite different. As a whole, the gross structure can be divided into 3 lobes, namely, spleen lobe, duodenal lobe and connecting lobe. According to the size, it can be divided into 3 types: the standard type, the thin type and the thick type ( among the 24 pigs, 3 types account for 16 / 24, 6 / 24 and 2 / 24, respectively) . The relative weight of pancreas accounts for 0. 09% to 0. 15% of body weight. The islets of Bama minipigs are generally small, and most of them are ultra-small type which compose of endocrine cell clusters. The relative weight of islets account for 1. 52% to 4. 98% of the pancreas and β cells account for 0. 012% to 0. 024% of body mass.




Establishment and Improvement of Abdominal Heterotopic Heart Transplantation Model in Rats by Single Operator 

ZENG Shun, LIU Daxing

2021, 38(3):  70. 

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Abstract: Objective To study and improve the establishment process of rat abdominal heterotopic heart transplantation model, reduce the difficulty of operation, shorten the learning cycle, and make the model easy to establish for the study of transplantation immunity, pathology and pharmacology. Method Through the establishment of rat abdominal heterotopic heart transplantation model, the operation skills were summarized in the whole process. Result There were 30 cases in the formal experiment, and the survival rate was 96. 7%. One case died of stabbing the heart during the operation and failed to stop bleeding after opening. The average cold ischemia time was ( 24. 2 ± 5. 0) minutes and total operation time was ( 59. 8 ± 0. 0) minutes. Conclusion The modified operation method of rat abdominal heterotopic heart transplantation model can shorten the operation time, reduce the difficulty of operation, and improves the sucess rate of the operation.




Laboratory Animal Pathological Monitoring and Implementation Results 

LAI Mengyu, ZHOU Xiangmei, ZHAO Deming, YANG Lifeng

2021, 38(3):  75. 

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Abstract: Laboratory animals are the foundation and irreplaceable tools for biology and medicine science development, as the quality of which relate closely to reliability and veracity of researches, as well as safety of related products. Therefore, strict quality control of laboratory animals is imperative. As the golden standard for diseases diagnosis, pathology consisted of clinical pathology, anatomic pathology and histopathology, reflects health condition of laboratory animals comprehensively and systematically, plays a crucial role in their quality control. Furthermore, pathology offers solid bases to standardize management and guarantee the quality of laboratory animals.




Research Progress in Animal Models of Intervertebral Disc Degeneration 

ZHANG Xinyi, YANG Yanni, JI Xiaoyan, LI Hanxiao, LIU Xifang

2021, 38(3):  79. 

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Abstract: Intervertebral disc degeneration is a precondition for all kinds of spinal degenerative disease and pathological basis. It affects the health of the patients and reduces their quality of life, and at the same time brings burden to family economy and society. Although there are many studies on intervertebral disc degeneration at home and abroad, the causes of intervertebral disc degeneration are not very clear. It is important to study the mechanism and treatment of intervertebral disc degeneration by using related animal models. According to the current research status of animal models of intervertebral disc degeneration, the selection and modeling method of experimental animals were summarized.