Table of Content

25 October 2022, Volume 42 Issue 5

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Review of Frontier Literature

Recent Research Progress of the Mechanism of Adult-born Immature Granule Neurons Underlying Rapid Antidepressant Effect of Ketamine

Sheng WEI, Xiwen GENG, Feng ZHAO, Zifa LI, Hao ZHANG, Minghui HU

2022, 42(5):  377-383.  DOI: 10.12300/j.issn.1674-5817.2022.146

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Major depressive disorder is a serious disease that impacts human health worldwide. Traditional antidepressant medications are limited by the delayed onset of therapeutic effects and the unacceptable side effects. Recently, the rapid antidepressant effect of ketamine, which decreases depressive symptoms within hours, has been a very attractive finding in the field, but the underlying mechanisms mediating this rapid response remain unclear. Besides, the clinical application of ketamine still carries risks, such as addiction and deleterious side effects. In May 2022, Rawat et al. reported that the acute activation of adult-born immature granule neurons (ABINs) in the dentate gyrus played a key role in the rapid antidepressant effects of ketamine. This commentary reviews the important findings of this research and looks forward to the future work.

Animal Models of Human Diseases

Changes in Expression of Monocarboxylate Transporters in the Rat Cerebral Cortex after Exercise-induced Fatigue Under Simulated High-altitude Hypoxia and its Significance

Chen GAO, Chunling FAN, Yurong LI, Wenjuan PEI, Caiping GUAN

2022, 42(5):  384-392.  DOI: 10.12300/j.issn.1674-5817.2022.050

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Objective To explore the correlation between adaptation to exercise-induced fatigue and lactate transport and metabolism in the cerebral cortex under high-altitude hypoxia. Methods A total of 63 SD rats were randomly divided into control, normal exercise, rush-entry-into-altitude, 3-day-altitude acclimatization, 1-week-altitude acclimatization, 2-week-altitude acclimatization, and monocarboxylate-transporter-inhibitor (altitude inhibitor injection) groups. Except for the control group, rats were subjected to exhaustive-load treadmill exercises under either normal pressure and normoxic conditions or low pressure and hypoxic conditions. The average time-to-exhaustion was used to determine variation trends in exercise-induced fatigue. The expression of monocarboxylate transporters (MCT)2 and MCT4 in the cerebral motor cortex was detected using western blotting and immunohistochemistry. The pathological evaluation of neuronal death in the cortex was carried out using Nissl staining, and the lactate content was also determined in rat brains. Results The average time-to-exhaustion in the rush-entry-into-altitude and the altitude inhibitor injection groups were (61.00±6.55) min and (71.25±9.59) min, respectively, which was significantly lower than that in the normal exercise (124.75±9.36) min and the 2-week-altitude acclimatization groups (100.25±9.74) min (P<0.05). Western blotting showed that, compared with the control group, the expression of MCT2 in the motor cortex in the 2-week-altitude acclimatization and the altitude inhibitor injection groups increased significantly by 120.6% and 164.4% (P<0.05), respectively; the expression of MCT4 in the two groups increased significantly by 174.6% and 168.8% (P<0.05). The results of western blotting were confirmed by the results of immunohistochemistry. In the high-altitude environment, the average lactate content in rat brains was significantly higher than that in the normal exercise group (0.175±0.021) mmol/g, P<0.05]. Neuropathological evaluation showed that the average neuronal density [neurons per high power field (HPF)] in the motor cortex in the rush-entry-into-altitude and the altitude inhibitor injection groups were (46.75±8.65) cells/HPF and (63.50±7.65) cells/HPF, respectively, which were significantly lower than that in the control group [(135.88±8.59) cells/HPF](P<0.05). Differences between the 2-week-altitude acclimatization [(121.75±16.00) cells/HPF] and the control groups were not statistically significant (P > 0.05). Conclusion The adaptability of the body to exercise fatigue after altitude acclimatization is correlated with changes in the expression of MCT2 and MCT4 in the brain, which can be used as targets for medical intervention for exercise-induced fatigue.

The Capacity of Silkworm Cocoon Water to Mitigate the Level of Oxidative Stress in Aged Rats

Huiyan QIN, Huafeng CHEN, Hui YANG, Hailan LUO, Weizhong FU, Qingbo LI, Jiehong ZHANG

2022, 42(5):  393-400.  DOI: 10.12300/j.issn.1674-5817.2022.027

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Objective To investigate the capacity of silkworm cocoon water (SCW) to mitigate the level of oxidative stress in aged rats. Methods The samples were processed by simulating the traditional SCW production process, and the SCW components were examined. The 16-month-old rats were randomly divided into 3 experimental groups that were treated with different doses of SCW, and 1 negative control group according to serum malonaldehyde (MDA) content, with 10 rats in each group. Experimental groups were fed SCW at doses of 500, 250, 125 mg/kg body weight, respectively, and the negative control group was given pure water, once a day, for 30 days. At the end of the experiment, the contents of lipid oxidation products (MDA), protein oxidation products (carbonyl), antioxidant enzyme activities (SOD, CAT, GSH-Px), and antioxidant substances (GSH) were measured. Results The crude protein content of SCW was 1 640 mg/100 g, the free amino acid content was 18 mg/100 g, and the hydrolyzed amino acid content was 1 700 mg/100 g. Serine (30.59%), aspartate (18.82%), glycine (10.00%), and threonine (8.24%) were the main hydrolyzed amino acids in SCW. Compared with the negative control group, SCW reduced the contents of MDA and protein carbonyl in the serum and tissue (P < 0.05), but increased the contents of GSH in the serum and tissue, the activity of SOD in tissue, and the activity of CAT and GSH-Px in the serum and tissue of aged rats (P < 0.05). Conclusion SCW has the effect of improving oxidative stress in aged rats.

Establishment of a Rabbit Heart-failure Model Using Transverse Aortic Constriction

Qingyi LUO, Tiling ZHANG, Yunchuan DING, Jian CHEN, Li ZHAO, Qinghui WANG

2022, 42(5):  401-408.  DOI: 10.12300/j.issn.1674-5817.2022.020

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Objective To investigate the method and feasibility of establishing a rabbit heart-failure model using transverse aortic constriction (TAC). Methods Twenty-five healthy male Japanese white rabbits were randomly divided into an operation group (n=15) and a sham-operation group (n=10). The animals in the operation group underwent TAC to establish the rabbit model of heart failure. The sham-operation group underwent thoracotomy but no narrowing operation. Cardiac function was evaluated using ultrasound at 4 and 8 weeks after operation. Eight weeks after operation, the descending aortic flow velocity was detected using cardiac ultrasound, combined with molecular biological indexes and pathological observations to verify the feasibility and stability of TAC in establishing the rabbit heart-failure model. Results Eight weeks after operation, 13 rabbits in the operation group and 10 in the sham-operation group survived. The blood flow velocities at the constriction in the operation group were >2.20 m/s. Compared with the sham-operation group, the following outcomes in the operation group increased significantly (P<0.05): heart rate, ascending aortic diameter, left atrial end-diastolic diameter, left ventricular end-diastolic diameter, and left ventricular end-systolic diameter. Left ventricular ejection fraction decreased significantly (P<0.05). Compared with the sham-operation group, the contents of left ventricular N-terminal pro B type natriuretic peptide (NT-proBNP) and β-myosin heavy chain (β-MHC) significantly increased in the operation group (P<0.05). Pathological sections showed that 8 weeks after TAC, cardiomyocytes in the operation group were hypertrophic and disordered, and myocardial fiber bundles widened. Interventricular septal end-diastolic thickness and left ventricular posterior wall end-diastolic thickness significantly increased in the operation group (P<0.05), compared to those in the sham-operation group at 4 weeks. Conclusion It is effective and feasible to establish an experimental rabbit heart-failure model using TAC. Cardiac ultrasound is an effective method to evaluate and determine the degree of aortic arch constriction and the success of heart-failure modeling. Rabbits in the operation group showed myocardial decompensation at 8 weeks after TAC, resulting in irreversible heart failure.

Measurement and Analysis of Bone Mineral Content and Bone Mineral Density in Healthy Cynomolgus Monkeys at Different Ages

Zhumei WEI, Guo SHEN, Zhenming LI, Yong ZENG, Feng JI, Jihong YANG

2022, 42(5):  409-415.  DOI: 10.12300/j.issn.1674-5817.2022.043

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Objective To study the skeletal characteristics of cynomolgus monkeys at different ages, and to provide reference values for skeletal research using cynomolgus monkeys as model animals. Methods A total of 283 cynomolgus monkeys aged 1-19 years were selected and divided into 8 groups according to sex and age :1 year≤age<3 years, 3 years≤age<5 years, 5 years≤age<7 years, 7 years≤age<9 years, 9 years≤age<11 years, 11 years≤age<13 years, 13 years≤age<15 years, and age≥15 years groups. Bone mineral density (BMD) and bone mineral content (BMC) were measured using dual-energy X-ray absorptiometry (DEXA). Results BMD increased from 0.32 g/m² to 0.57 g/m² and BMC increased from 67 g to 399 g in male cynomolgus monkeys between the age of 1 and 12. While BMC and BMD were relatively stable in the 12~15 years group. BMC and BMD remained at (367.51±7.17) g and (0.56±0.06) g/m², respectively, in the ≥15 year-old group. In female cynomolgus monkeys, BMC increased from 58 g to 233 g and BMD increased from 0.31 g/m² to 0.47 g/m² between the age of 1 and 10. The maximum BMC was only about 58% of that of male cynomolgus monkeys, the maximum BMD was about 80% of that of male cynomolgus monkeys, and BMC decreased significantly after the age of 10 years. BMC remained at (166.63±6.21) g and BMD remained at (0.46±0.04) g/m² after the age of 15 years, which were 45% and 80%, respectively, of those of male cynomolgus monkeys. Conclusion In male cynomolgus monkeys, as their skeleton develops, BMC and BMD increase continuously until the age of 12 years, and are relatively stable between the age of 12 and 15 years. While in female cynomolgus monkeys, BMC and BMD increase annually until the age of 10 years, but after the age of 10 years, BMC decreases significantly, while BMD remains relatively stable between the age of 10 and 15 years. Peak bone mass of female cynomolgus monkeys is lower than that of male cynomolgus monkeys.

Effect of Dexamethasone on Cartilage Degeneration in Rats with Collagen-induced Arthritis

Bingxin XU, Kaijian FAN, Tingyu WANG, Huijin CHEN

2022, 42(5):  416-422.  DOI: 10.12300/j.issn.1674-5817.2022.066

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Objective To establish a rat model of collagen-induced arthritis (CIA), a rat model of rheumatoid arthritis, and to study the protective effect of dexamethasone (DEX) on cartilage in CIA rats. Methods Thirty Wistar rats were randomly divided into three groups: normal, model, and DEX groups. The CIA model was established by intradermal injection of a white emulsion mixed with 0.1 mL bovine type II collagen and Freund's incomplete adjuvant in equal proportions into the distal part of the rat tail. The dates of the two injections were recorded as days 0 and 7, respectively. After the establishment of the CIA model, DEX (1 mg/kg) was intraperitoneally injected in the treatment group 3 times a week for 4 weeks. After the administration, the rats were euthanized, and the knee cartilages of 10 rats in each group were divided into three parts. The first part was fixed, decalcified, and stained with Alcian blue to observe the pathological changes in the cartilage layer. The second part was used for detection of mRNA expression of matrix metalloproteinase (MMP)-9 and MMP-13, and a disintegrin-like and metalloproteinase with thrombo-spondin motifs (ADAMTS)-4 and ADAMTS-5 by using real-time quantitative PCR analysis in the femoral head cartilage. The third part was used for detection of the expression levels of MMP-9, MMP-13, and ADAMTS-5 using immunohistochemistry in the cartilage layer of knee joint. Results Compared with the model group, the decrease of cartilage area, cartilage thickness, and cartilage cell numbers were significantly inhibited after DEX treatment (P<0.05). The cartilage surface was smooth and the invasion degree significantly improved, which indicated that cartilage destruction was inhibited. After DEX treatment, the mRNA levels of MMP-9, MMP-13, ADAMTS-4, and ADAMTS-5 in the cartilage of CIA rats were decreased (P<0.001), and the protein expression levels of MMP-9, MMP-13, and ADAMTS-5 were also significantly decreased (P<0.05). Conclusion DEX can inhibit the expression of various MMP in CIA rats and has a therapeutic effect on cartilage destruction.

Research Progress on Animal Models of Intrauterine Growth Restriction

Qiwen HU, Zheng BI, Haiping LIU, Zhihua DONG, ZHUYanlin, Jinhua WANG

2022, 42(5):  423-431.  DOI: 10.12300/j.issn.1674-5817.2022.063

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The occurrence of intrauterine growth restriction (IUGR) may be related to maternal malnutrition, abnormal placental function, immune abnormalities, genetically related problems as well as other diseases, but the mechanism is still unclear. Therefore, the study of IUGR and the development of its animal model are critical issues in obstetrics. IUGR models are mainly based on laboratory rodents, such as mice and rats, and other mammals such as pigs, rabbits and sheep. This article introduced several common IUGR animal models, including nutrition restriction model, high-altitude pregnancy model, natural selection model, and nicotine exposure model, and also described the construction methods of IUGR models and the comparison of their advantages and disadvantages.

Characteristics and Application of Transgenic Mouse Models in Alzheimer's Disease

Feng WEI, Weiwei CHENG, Yafu YIN

2022, 42(5):  432-439.  DOI: 10.12300/j.issn.1674-5817.2021.182

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Alzheimer's disease is a common neurodegenerative disease characterized by progressive memory loss and overall cognitive decline. Two typical pathological features are senile plaques and neurofibrillary tangles detected in the brain, which consist of large amounts of precipitated amyloid peptide and hyperphosphorylated Tau protein, respectively. In addition, there are other pathological features such as neuroinflammatory response and extensive loss of neurons. In the past decades, a lot of research has been done on Alzheimer's disease, but its etiology and pathogenesis are still unclear, and there is no ideal treatment or radical cure. This review introduced the characteristics of various transgenic animal models and their current application in preclinical studies, in order to provide theoretical basis for future experimental studies.

Facilities and Management for Laboratory Animals

Preliminary Study on Raising and Maintaining Clean Mice with Micro-barrier Cages in Conventional Environmental Facilities

Qi REN, Yandong LI, Qiang SUN

2022, 42(5):  440-447.  DOI: 10.12300/j.issn.1674-5817.2022.129

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Objective To explore the feasibility of breeding cleaning degree mice with micro-barrier cages in conventional environmental facilities. Methods Clean-grade mice were housed and maintained in positive-pressure microbarrier cages in a general environment, during which third-party sampling of the microbial status of the animals in the microbarrier was performed on a quarterly frequency. Results From June 2020 to July 2022, no microorganisms that must be excluded by the national standard for cleaning degree mice have been detected in two consecutive years. Conclusion Micro-barrier cages can be used to raise and maintain cleaning grade mice in conventional environmental facilities.

Application of the Wechat Booking Platform in Reservation System of the Operating Room in Laboratory Animal Center of Beijing Friendship Hospital

Lei WANG, Hufeng XU

2022, 42(5):  448-452.  DOI: 10.12300/j.issn.1674-5817.2022.017

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In order to effectively improve the fine management level of animal experimental operating room and ensure the smooth development of animal experimental research, operating room of Experimental Animal Center of Beijing Friendship Hospital has established a WeChat reservation platform. The operating room implements an appointment system, and pushes the appointment information to the experimental participants through the WeChat platform, so that the appointment information can be publicized in a timely and effective manner. The platform provides a standardized operating room application procedure for experimental participants in relevant departments. Through the analysis of appointment information, platform managers can timely understand and grasp the focus and needs of animal experimental operating room management. This paper analyzed and summarized the running condition of WeChat reservation platform in the past one year, in order to promote the communication and discussion of related problems.

Discussion on the Construction Experience of Laboratory Animal Facilities in Universities and Research Institutes

Huiran LIN, Zhaoxia LIU, Xiaobiao FENG, Minjie LIU, Xiaojie HUANG

2022, 42(5):  453-457.  DOI: 10.12300/j.issn.1674-5817.2022.042

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Laboratory animal facilities, as the main place for animal raising and experiment, have strict requirements on environmental conditions such as temperature, humidity, pressure, noise, air cleanliness class and ventilation rate. Laboratory animal facilities also have standardized management requirements for personnel operation, goods and animals entering and leaving the facilities. Based on the construction and management experience of laboratory animal facilities in Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences in the past 10 years (2012-2022), we summarized and discussed the experience from design planning to equipment selection, aiming to provide practical reference of the construction of experimental animal facilities to universities and research institutes.

Investigation and Discussion on the Standardization Work of Feeds for Laboratory Animals

Jianbo LUO, Junhui LI, Jia ZHOU, Yong HE, Min FU

2022, 42(5):  458-465.  DOI: 10.12300/j.issn.1674-5817.2022.019

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Standardization of laboratory animals is the basis for obtaining accurate and stable experimental data, drawing objective and reliable experimental conclusions. The standardization of laboratory animal feeds is an integral part of laboratory animal standardization and an important support to realize laboratory animal standardization. In this paper, we compared the historical process of laboratory animal feeds standardization at home and abroad, outlined the key problems existing in the current domestic work, and put forward targeted countermeasures in order to provide references for the standardization of laboratory animal feeds in China.

Educational and Teaching Practice

A Novel Model for in-vivo Training of Postgraduates on Cardiac Electrical Conduction System Mapping Skills

Xiaokang WANG, Ruojin ZHAO, Yunhui LÜ, Guangxin YUE, Shangyu LIU, Ting HE, Peng PENG, Liang MENG, Jubo LI, Baojie ZHANG, Chen SHEN, Yongchun CUI, Xin WANG

2022, 42(5):  466-471.  DOI: 10.12300/j.issn.1674-5817.2022.059

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Objective StandardizationTo explore a novel special technique intensive mode (STIM) of teaching cardiac conduction system mapping skills. This method includes theoretical lecture, in vivo operation, 5-whys methods of analyses, intensive-problem oriented trainings, and assessments. Methods Ten post-graduates majoring in cardiology who have equivalent education backgrounds and technical levels were randomly divided into two groups (n=5 per group). All participants underwent a skill training on cardiac electrophysiology mapping, with one group training under the STIM and the other group under the traditional "theory-practice-test" (TPT) mode. The effect of these two educational techniques were evaluated at the end of the training. Results Postgraduates in the STIM group had a more solid grasp of the anatomical position and structure of the cardiac conduction system than those in the TPT group. During in-vivo operation, the students in the STIM group reconstructed and interpreted the electroanatomical maps, voltage maps, electric excitation maps, and electric conduction maps more accurately and quickly than those in the TPT group. Conclusion The STIM teaching mode more effectively improved the basic surgical skills of postgraduates, stimulated the students' learning initiative, and improved the quality of education. STIM is a kind of intensive teaching mode of special skills, and is worth widely implementing in the education of postgraduates majoring in cardiovascular diseases.

Thinking of Teaching Reform in the Animal Experiment of Operative Surgery in Medical School

Yufei LENG, Wenjing LU, Fengwei WANG

2022, 42(5):  472-476.  DOI: 10.12300/j.issn.1674-5817.2022.029

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Animal experiment of operative surgery is a compulsory course for medical students and an important basic course for surgical practice teaching, which is of great importance to the cultivation of clinical medical talents. Combining the teaching practice experience of Shanghai Jiao Tong University School of Medicine, this paper analyzed the common problems in domestic medical universities, and put forward the optimized measures including adjusting and enriching teaching contents, expanding and enriching teaching resources, improving innovative teaching models, constructing scientific evaluation system, and strengthening the construction of teaching faculty, so as to improve the quality of the course and the comprehensive quality of medical students, help cultivate excellent medical talents, and provide references for the teaching reform of animal experiment of operative surgery in medical colleges and universities.