Table of Content

28 June 2024, Volume 41 Issue 3

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Study on the Protective Effect and Mechanism of Monoester Alkaloidson Cardiomyocytes of CHF Rats

ZHAO Qianqian, WANG Shen, LU Xiangqin, YANG Ting

2024, 41(3):  1-8.  DOI: 10.3969/j.issn.1006-6179.2024.03.001

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e To analyze the protective effects of single ester alkaloids on myocardial cells ofchronic heart failure rats and their effects on PI3K / AKT pathway. Method 100 rats were randomlydivided into group A ( blank group) , group B ( model group) , group C ( low-dose group) , group D实验动物科学 41 卷(medium-dose group) and group E ( high-dose group) with 20 rats in each group. Groups C, D and Ewere given aconite concentrate by intragastric administration after successful modeling of chronic heartfailure model, and the doses were 2. 79, 5. 57 and 11. 14 g / kg, respectively. After 6 weeks of gavage,histopathological changes, echocardiographic examination indexes, myocardial cell apoptosis rate,expression of cardiac tissue proapoptotic protein ( Bax ) , B-lymphoblastoma-2 ( Bcl-2) , Cagpase-8,autophagy protein, BNP, AI of myocardial cells in each group were observed. Result Theechocardiography indexes IVSs, IVSd, LVPWs, LVPWd, the apoptosis rate of cardiomyocytes, theexpression of Bax, Caspase-8, the levels of IL-6, TNF-α, the expression of autophagy protein LC3-Ⅱ,LC3-Ⅱ / Ⅰ, BNP and AI in cardiomyocytes in groups E, D and C were higher than those in group A.LVEF, expressions of Bcl-2, PO2 , autophagy protein P62, p-PI3K and p-AKT in groups E, D and Cwere lower than those in group A (P<0. 05) . The IVSs, IVSd, LVPWs, LVPWd, the apoptosis rate ofmyocardial cells, the expression of Bax and Caspase-8, levels of IL-6, TNF-α, autophagy protein LC3-Ⅱ, LC3-Ⅱ / Ⅰ, BNP and AI in groups E, D and C were lower than those in group B. LVEF, theexpressions of Bcl-2, PO2 , autophagy protein P62, P-PI3K and P-Akt in groups E, D and C were higherthan those in group B (P<0. 05) . With the increase of concentration of monkshood concentrate, IVSs,IVSd, LVPWs, LVPWd, myocardial cell apoptosis rate, Bax, Caspase-8 expression, IL-6, TNF-αlevels, myocardial autophagy protein LC3-Ⅱ, LC3-Ⅱ / Ⅰ, BNP and AI levels decreased. LVEF, theexpressions of Bcl-2, PO2 , autophagy protein P62, p-PI3K and p-AKT in myocardial cells wereincreased (P < 0. 05 ) . Conclusion Single ester alkaloids may activate PI3K / AKT pathway by upregulating the expression of p-PI3K and p-AKT, and alleviate the malignant progression of heart failure ina dose-dependent manner.

Study on Behavioral Evaluation Method for Establishing InsomniaModel with Para-chlorophenylalanine

QU Zihan, MA Haizhao, ZHANG Ye, LIU Yinghao, ZHI Xiaoyu, LI Baojie, ZHANG Hongshi

2024, 41(3):  9-13.  DOI: 10.3969/j.issn.1006-6179.2024.03.002

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By observing the changes in learning and memory abilities in various behavioralexperiments of the para-chlorophenylalanine ( PCPA) insomnia rat model within a week after successfulmodeling, the stability of the PCPA insomnia model was verified. Method Thirty SD rats were randomlydivided into blank group and model group, the rats in the blank group were fed normally, andestablishment of insomnia model by intraperitoneal injection of PCPA into model group rats. Fivebehavioral tests including Morris water maze test were performed in the first week after modeling.Result Compared with the blank group, the sleep latency of the model group was significantly prolonged(P < 0. 01 ) , and the sleep duration was significantly shortened ( P < 0. 01 ) . In the water mazeexperiment, the time of seeking platform in the model group was prolonged ( P < 0. 01) . In the Barnesmaze experiment, the total residence time at the target portal of the model group rats during the testexploration period was significantly shortened (P<0. 01) . In the dark avoidance experiment, the numberof errors in the model group increased ( P < 0. 05) , and the latency was significantly prolonged ( P <0. 01) . In the elevated cross maze experiment, the open arm residence time of rats in the model group实验动物科学 41 卷was significantly shortened ( P < 0. 01 ) . In the open-field experiment, the dwell time, the number ofuprightness and the total distance of movement in the central area of the model group were all lower thanthose of the blank group, among which the dwell time in the central area was significantly different (P<0. 01) . Conclusion The PCPA insomnia model has high reproducibility and stability, and can be usedas a basic model for sleep and memory-related studies.

Application of Water Exchange Method inColonoscopy Bowel Preparation in Pigs

CHEN Zhong, DONG Jun, LI Dan, YIN Jiuyong, LIAO Feifei, LU Yali, YAN Yahui, ZHAO Tao

2024, 41(3):  14-17.  DOI: 10.3969/j.issn.1006-6179.2024.03.003

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Objective Explore a safe and effective bowel preparation method to improve the quality ofpreparation in pigs colorectal model. Method 40 cases of endoscopic treatment of experimental pigsunderwent long transparent cap associated water exchange method in colonoscopy to flush feces or onlyfasted without any other treatment in the intestinal tract of pig for bowel preparation. The time of bowelpreparation, the degree of bowel preparation, and the degree of satisfaction of the endoscopist would berecorded. Result The cleaning preparation was satisfied, the operation field was clear and achieved theexperimental requirements. Conclusion It has high application value of long transparent cap combinedwith water exchange method in colonoscopy in preparation for operation in the experimental pig colon.

Pathological Evaluation of Skin Injury and Healing by Different Modeling Methods

XIAO Sa, SUN Jun, PENG Dongdong, TANG Hailiang, JIANG Dejian, ZHANG Miaohong

2024, 41(3):  18-25.  DOI: 10.3969/j.issn.1006-6179.2024.03.004

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Objective To investigate the effects of different method of modeling on skin injury and thepathological evaluation of wound healing in different time periods. Method Seventy-eight SD rats, halfmale and half female, were divided into normal control group ( 14 rats) , burn model group ( 24 rats) ,scald model group (24 rats) and total skin resection group (16 rats) . In the scald model group, the skintissue was scalded by desktop super temperature control scald instrument, and the mixed bacteria weredripped into the wound surface to establish the model of superficial II degree scald combined with woundinfection. Burn model group was established by using self-made burn conical flask to burn skin tissue anddripping mixed bacteria into the wound surface to establish the model of superficial II degree burncombined with wound infection. In the total skin resection model group, the skin was resected with asterile scalpel in an area of 3 cm × 3 cm without muscle injury to establish the total skin resection model.第 3 期 肖 洒等:不同造模方法对皮肤损伤及愈合情况的病理学评价Two animals of the normal control group were dissected and the samples were taken on the day 1, 3, 4,5, 7, 10 and 14 respectively. Four animals of the burn and scald + bacterial infection model group weredissected and the samples were taken on the day 1, 3, 5, 7, 10 and 14 respectively. Four animals of thetotal skin resection model were dissected respectively on the day 4, 7, 10 and 14. The skin tissues of thewound were examined pathologically. Result The gross observation showed that the skin of differentmodels had different degrees of wound damage at the initial stage of modeling; in the middle stage, localreactions such as redness, swelling and blisters appeared around the skin injury site; the final skinhealing is relatively complete. Microscopic observation showed that the skin of each model in juniormiddle school was mainly characterized by epidermal degeneration and necrosis, incomplete keratosis,lack of granular layer, acanthocyte layer and basal cell layer, and different degrees of inflammatory cellinfiltration in the dermis. At the end of the period, the structure of each layer of skin epidermis isrelatively complete, and a large number of new blood vessels, fibroblasts and a small amount ofinflammatory cells can be seen in the dermis. Conclusion Through the pathological evaluation of skintissue at different stages of modeling, the injury and healing of skin caused by different modeling methodwere comprehensively analyzed, which laid the foundation for the follow-up pharmacodynamic study.

Establishment of Genetic Quality Testing and Strain Identification SNPpanel for Six Classes Inbred Rats

ZHANG Xiaoqing, WANG Yan, XIE Fei, CHEN Hong, ZHAN Dawei

2024, 41(3):  26-32.  DOI: 10.3969/j.issn.1006-6179.2024.03.005

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Objective To screen out single nucleotide polymorphism ( SNP ) loci evenly distributed onthe rat chromosome that can be used to establish the genetic quality detection and strains identification ofcommon inbred rats. Method A total of 141 SNPs were obtained from the literatures, and theinformation obtained in the Ensembl database was used to screen out SNPs with good polymorphism in 6rat strains of BN, F344, GK, LEW, SHR and WKY. From the above SNPs, the SNP combination thatevenly distributes each chromosome and contains strain-specific information was optimized. The effect ofcombined SNPs in genetic quality detection and strain identification of common inbred rats was verified byPCR amplification technology and Sanger sequencing method. Result 40 SNP markers were optimized,which were monomorphic within each strain and polymorphic among strains. They contain specific SNPsthat can be used for strain identification, including 4 SNPs for BN rats, 3 SNPs for F344 rats, 2 SNPs forGK rats, 2 SNPs for LEW rats, and 1 SNP for SHR rat. The optimized 40 SNPs were verified by usingDNA mixed samples of BN rats, F344 rats, LEW rats, SHR rats and WKY rats respectively, and the第 3 期 张晓晴等:建立 6 种近交系大鼠遗传质量检测及品系鉴别的单核苷酸多态性组合result were consistent with the website data. Conclusion A SNP panel for genetic quality detection andstrain identification of common inbred rats was successfully established

Exploration of the Effect of Specific Immunity on Inflammatory Response inAsthmatic Mice Bronchoalveolar Lavage Fluid Based on Th1 / Th2 Expression

TAO Jun, KE Jing, CHEN Lin, CHEN Zhuo, XIE Huihui, LI Jinquan

2024, 41(3):  33-39.  DOI: 10.3969/j.issn.1006-6179.2024.03.006

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Objective To explore the effect of specific immunity on inflammation in bronchoalveolarlavage fluid ( BALF) of asthmatic mice by analyzing Th1 / Th2 expression. Method Forty mice weredivided into control group, model group, SIT group, and dexamethasone group, with 10 mice in eachgroup. Except for the control group, asthma mouse models were established in the other groups. After thestimulation phase, the SIT group was injected with 1 mg of OVA, the dexamethasone group was given a0. 075 mg / mL solution, and the remaining rats were injected with an equal volume of PBS solution. Thelung function meter was used to detect airway responsiveness; the BALF inflammatory cells were observedby Wright-Giemsa staining; ELISA was used to detect cytokines IL-13, IL-4, IL-5, and INF-γ in BALF;HE staining was used to observe lung tissue morphology; PAS staining was used to observe cup-shapedcell activity in lung airways; and flow cytometry was used to detect the proportion of Th1 and Th2 in lungtissues. Result Specific immunity significantly reduced airway hyperreactivity in model mice, reducedinflammatory cell activity in BALF, and inhibited asthma occurrence and development by increasing Th1实验动物科学 41 卷while decreasing Th2. Conclusion Specific immunity can alleviate airway hyperresponsiveness andinhibit inflammation in asthmatic mice, improve cup-shaped cell proliferation, and relieve ventilation,possibly related to regulating Th1 / Th2 expression.

Effects of Different Routes of Enrofloxacin Administration on Body Weight andBlood Physiological and Biochemical Indexes of Guinea Pigs

DONG Hao, LIU Zhiguo, LIU Jinsheng, WU Weiguo, WANG Xuewen, ZHANG Leying, DONG Qinghua, ZHU Wanyue, MA Liying, ZUO Qin

2024, 41(3):  40-45.  DOI: 10.3969/j.issn.1006-6179.2024.03.007

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Objective To study the effects of different routes of enrofloxacin administration on bodyweight and blood physiological and biochemical indexes of guinea pigs, this study was performed.Method The effects of different routes of enrofloxacin administration on body weight,blood physiologicaland biochemical indexes of guinea pigs were determined respectively. Statistical method were used fordata analysis. Result High dose of enrofloxacin administered by drinking water significantly affected thewater consumption of guinea pigs. Both high dose of enrofloxacin and low dose of enrofloxacinsignificantly affected the weight gain of guinea pigs. Compared to the control group, several significantlydifferent blood physiological indexes were determined in the administration treatment groups includingWBC, MON, GRAN, MCHC, RDW-CV, PDW. In terms of blood biochemical indexes, five parameters(Ca, ALT, ALP, γ-GT, and TG) were also affected after administration, compared to the control第 3 期 董 浩等:不同途径给药恩诺沙星对豚鼠体质量和血液生理生化指标的影响group. Conclusion The determination of the effects of different routes of enrofloxacin administration onseveral biological indexes of guinea pigs would provide basic data for scientific medication in guinea pigs.

Effects of Three Kinds of Drinking Water on Blood Indexes in NIH Mice

WANG Xuewen, WU Zhenghong, PENG Weijuan, QIU Liyuan, DONG Hao, ZUO Qin, LIU Zuomin

2024, 41(3):  46-50.  DOI: 10.3969/j.issn.1006-6179.2024.03.008

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Objective To explore the effects of three kinds of drinking water on blood indexes in NIHmice. Method 54 NIH mice were randomly divided into three groups, drinking autoclaved water,autoclaved acidified water and sterile water bag water ( membrane filtered water) , respectively. On the30th, 60th, and 90th days of the experiment, blood was taken for blood routine and biochemical tests,and liver tissue was taken for histopathological examination to explore the effects of three kinds of drinkingwater on the health status of mice. Result The albumin ( ALB) in the 60 day water bag group wassignificantly higher than that in the autoclaved water group ( P < 0. 05) , the total protein ( TP ) in theautoclaved acidified water group was significantly higher than that in the autoclaved water group ( P <0. 01) , and there was no significant difference between the other groups ( P > 0. 05 ) . The liverhistopathological result showed that no pathological changes were found in the liver tissues of mice in eachgroup, and they were normal liver tissues. Conclusion Three kinds of drinking water have no significantimpact on the health status of NIH mice.

Effect of Different Dehydration and Wax Immersion Procedures on theQuality of Paraffin Sections of Mouse Liver and Intestine

WU Haifeng, ZHOU Xiaojiang, WEI Juanpeng, LYU Ming, GAO Ming

2024, 41(3):  51-54.  DOI: 10.3969/j.issn.1006-6179.2024.03.009

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Objective To compare the effects of two dehydration procedures and five wax immersion timeson the quality of paraffin sections of mouse liver and intestinal tissues, with a view to optimizing thepreparation procedures of paraffin sections. Method The liver and intestinal tissues of BALB / c micewere prepared in two different dehydration procedures ( Procedure I and Procedure II) and five differentwax immersion procedures ( total immersion time of 90, 120, 150, 180 and 210 min) to compare theresult of tissue sectioning and HE staining. Result The paraffin sections produced by dehydrationprocedure 1 were easily fragmented, with obvious wrinkles and cavities in the center of the tissue; theparaffin sections produced by dehydration procedure 2 were intact and free of cavities, and the structureof the paraffin sections was observed to be intact and clear under the microscope after HE staining.Compared with the other four wax immersion procedures, the paraffin sections produced with a totalimmersion time of 150 min were the most complete and clear after staining. Conclusion The quality ofthe paraffin sections produced by using the dehydration procedure II and the total time of wax immersionof 150 min was improved and easy to observe under the microscope, which is recommended as theoptimized procedure for paraffin sections of mouse liver and intestinal tissues.

Constructive Expression of Creatine Kinase Isoenzyme CK-MB andAntibody Production and Preliminary Validation

LI Linyu, YAO Xinxin, LIU Junling, TAO Qianqian, YANG Juan

2024, 41(3):  55-64.  DOI: 10.3969/j.issn.1006-6179.2024.03.010

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Objective Construct heterodimeric CK-MB recombinant protein and prepare its specific geneengineered antibody as the diagnostic raw material of CK-MB quantitative detection kit, so as to assist theearly diagnosis of acute myocardial infarction in clinic, and to improve the accuracy of diagnosis. MethodMouse plasma cells were obtained by obtaining heterodimeric CK-MB through co-expression vector andanimal immunization, further, single mouse B cells specific for antigen were isolated using single-cellphotoconductive system, and genetically engineered antibodies were constructed after obtaining the genesequences of the variable regions of the light and heavy chains of the antibodies through single-cell PCR.Candidate antibodies were screened based on preliminary functional validation, and the optimal pairingscheme was further explored by checkerboard titration, and a CK-MB quantitative detection kit wasinitially assembled using the principle of double-antibody sandwich ELISA. Result CK-MB immunogenwas successfully obtained; single plasma cells specific for CK-MB antigen were screened by single B-cellphotoconductivity and two strains of mouse-derived genetically engineered antibodies were prepared, andboth of them had the ability of recognizing the natural CK-MB; two pairwise protocols were screened by double-antibody sandwich ELISA, and the R2of the fitted curves were both greater than 0. 99.Conclusion Mouse-derived genetically engineered antibody targeting CK-MB was successfully prepared,and the specificity and affinity of the antibody was preliminarily verified by ELISA, laying a solidfoundation for the further development of a new type of CK-MB quantitative detection kit with highspecificity and high sensitivity.

Handing and Thinking for the Occasional Diseased Animals in Scientific Research

ZHANG Nana, ZHANG Zhixuan, FENG Zhe, ZHANG Yuchi, WANG Rongfeng, WANG Kailing, YU Jiawei, LYU Yi

2024, 41(3):  65-68.  DOI: 10.3969/j.issn.1006-6179.2024.03.011

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Objective We summarized and analyzed the scientific thinking and related treatment methodcaused by occasional sick large animals in scientific research to provide practical reference to handle thesimilar conditions. Method In a study of techniques related to laparoscopic porcine livertransplantation, the liver donor pig developed a sudden respiratory cardiac arrest after routine anesthesiavia the marginal ear vein. Therefore, the cyst wall cells were cultured and the cyst wall tissues wereexamined pathologically. Result The cyst wall cells showed a grape tandem suspension growth. The smear staining result showed that the cultured cyst wall cells were mainly inflammatory cells. The cystwall tissue was mainly composed of granulation tissue and fibrous tissue, which was a chronicinflammatory hyperplasia. Conclusion The following enlightenments can be obtained from the occasionalencounter of this sick pig: ①routine animal health examination should be performed before large animalexperiments; ②researchers should have a high sensitivity to infectious disease protection; ③researchersshould maintain a highly scientific sensitivity to the occassional situations during the experiment;④comprehensive pathological samples should be retained during the experiment; ⑤ the monitoring ofanimal health status in the farm should be improved

A New Method to Keeping Miniature Pigs in Supine Position

MO Ansheng, CHEN Lin, LIN Lianfang, WANG Guozheng, QIN Minghui, WANG Ruirui, GUAN Hanbin, ZENG Delan

2024, 41(3):  69-71.  DOI: 10.3969/j.issn.1006-6179.2024.03.012

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Objective To study a method of fixing Guangxi Bama miniature pig in supine position so as tofacilitate operation. Method Selected 12 mature Guangxi Bama mini pigs, which operation for pleuralorgan procurement, fixed in supine position with a special fixation device after general anaesthesia. ResultAll the pigs were well maintained in supine position during the operation without rolling over or shifting.Conclusion It is a simple and effective method to keep Guangxi Bama miniature pigs in supine positionwith the special fixation device.

Advances in Canine Tumor Detection Technology

LI Yanshan, WANG Zhouxiang, HU Yufeng

2024, 41(3):  75-78.  DOI: 10.3969/j.issn.1006-6179.2024.03.013

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With the continuous progress of society and the improvement of people’ s living standards, petownership has become an indispensable part of people’ s spiritual life. However, in reality, many petowners lack scientific pet-raising concepts, leading to missed optimal treatment opportunities for pets.Diseases may deteriorate into malignant tumors, even leading to the unfortunate demise of pets. Given theuncertainty of tumor onset time and disease progression, early screening and timely treatment areparticularly crucial. This article comprehensively discusses the commonly used detection techniques forcanine tumors in recent years, and delves into the practical applications, advantages, and disadvantagesof these techniques. This is significant for achieving targeted, accurate, and rapid diagnosis of caninetumors in clinical settings, and provides an important theoretical basis for rapid and accurate diagnosisand treatment of canine tumors.

Research Progress in Pathogenesis and in Vitro-in Vivo Model of Alzheimer’s Disease

LYU Shujie, GUO Wen, FANG Liang, ZHANG Caiyun

2024, 41(3):  79-84.  DOI: 10.3969/j.issn.1006-6179.2024.03.014

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Alzheimer’ s disease ( AD ) is a neurodegenerative disease characterized by cognitivedysfunction. As China’ s population aging becomes more and more serious, the proportion of peoplesuffering from AD is increasing, it is urgent to study its pathogenesis and establish appropriatepathological models to further study the prevention and treatment of AD. In this paper, in vitro and invivo AD models were reviewed, especially ten in vivo animal models (β-amyloid precursor protein (APP)transgenic model, D-galactose model, D-galactose combined with β-amyloid protein (Aβ) model and soon) and three in vitro models ( HT22 cell model, PC12 cell model and SH-SY5Y cell model) wereintroduced in detail. This study will provide AD reference and research ideas for further study and newdrug development for AD disease.

Comparison on Molecular Typing Technologies and the Infection Condition ofStaphylococcus Aureus in Laboratory Animals

WU Yunpu, ZHANG Zehui, CUI Ziyin, LIANG Xicai, WANG Xing, TA Mingjun, WANG Chuntian

2024, 41(3):  85-89.  DOI: 10.3969/j.issn.1006-6179.2024.03.015

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 Staphylococcus aureus ( S. aureus) is a zoonotic opportunistic pathogen, which is must bemonitored and excluded from the barrier environment and often is a serious threat to the quality oflaboratory animals and the health of practitioners. It is critical to use accurate, sensitive, and rapidmethod of molecular typing to control the transmission of S. aureus in the laboratory animal barrierenvironment. The molecular method for typing S. aureus main including Staphylococcal cassettechromosome mec typing, Staphylococcal protein A typing, multilocus sequence typing, Pulsed field gelelectrophoresis typing, accessory gene regulator typing, and others. Understand the infection status ofStaphylococcus aureus in laboratory animals of barrier systems and comparation of the technicalcharacteristics and applications of different molecular typing method, which can provide a reference forthe detection and typing of Staphylococcus aureus of laboratory animal origin.

Application of Vasopressor Activity Test in Drug Biological SafetyTests and Analysis of Experimental Key Points

ZHANG Yuan, GUO Longjing, WU Yanlin, YANG Ze’ an, NA Tao

2024, 41(3):  90-94.  DOI: 10.3969/j.issn.1006-6179.2024.03.016

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The vasopressor activity test is one of the drug biological safety test method included in theChinese Pharmacopoeia. It is used to detect whether the limits of vasopressor in APIs and relatedpreparations that may contain impurities which may raise blood pressure meet the standard requirements.Although the application of this test is narrow, it plays an important role in the safety of clinicaltreatment. This article summarizes the experimental principles, experimental systems, methodapplications, improvement suggestions, and experimental key points of the vasopressor activity test, sothat provides references for relevant experimenters.