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    25 April 2022, Volume 42 Issue 2
    Laboratory Animal and Biosafety
    Application Prospect of Identification and Traceability Technology of Laboratory Animals in Biosafety Laboratory
    Lixiang CHEN, Boyin QIN, Hua YANG, Chunhua XU, Xiuhua PENG, Shun LI, Xiaohui ZHOU
    2022, 42(2):  89-94.  DOI: 10.12300/j.issn.1674-5817.2021.174
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    Effective management of laboratory biosafety plays one of the most important roles in the national biosafety system. Laboratory animals have been widely used in the life science research field, including infection animal experiments, which have to be done in the animal biosafety laboratory (ABSL). The risk assessment and control related to laboratory animals and animal experiments are the most important affairs in ABSL, and the identification and traceability of laboratory animals detected by wireless tracking technology represent a developing tendency in ABSL management. From the perspective of biosafety, this paper summarized the development of laboratory animal identification and traceability technology,and described the application prospect in ABSL management of non-contact identification technology, especially radio frequency identification technology, which can provide reference for relevant practitioners to use the identification and traceability technology of experimental animals for biosafety control.

    Biosafety Problems and Countermeasures in the Management of Barrier Facilities for Laboratory Animals
    Yuqin YANG
    2022, 42(2):  95-101.  DOI: 10.12300/j.issn.1674-5817.2021.164
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    Biosafety is an important part of national security, and effective control of biological hazards of laboratory animals is the basis of using laboratory animals. Based on the experience of biosafety management in the Laboratory Animal Center of Shanghai General Hospital, this paper analyzed the potential biosafety threats faced by laboratory animal barrier facilities in the process of ensuring the smooth conducting of animal experiments from the perspective of a laboratory animal facility manager, and discussed the relevant countermeasures to reduce such risks, in order to provide a reference for the proper control of biosafety problems in animal experiments.

    Performance Test and Evaluation of Self-developed Biosafety Cage Changing Workbench
    Shibo SU, Wenjie MA, Wei WANG, Huijuan GUO, He ZHANG, Zi WANG, Lei KANG, Shouqi LIU, Huairan LIU, Changde SI, Chuntian GOU, Wei ZHANG, Hongyan CHEN
    2022, 42(2):  102-110.  DOI: 10.12300/j.issn.1674-5817.2021.102
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    Objective To test the function and evaluate the safety of the self-developed biosafety cage changing workbench.Methods According to the criteria of YY 0569-2011 "Class II Biological Safety Cabinet" and RB/T 199-2015 "Laboratory Equipment Biosafety Performance Evaluation Technical Specifications", we detected the various performance indicators of the cage changing workbench (HT-1303), which was independently developed and produced by Shenzhen Hongteng Biological Technology Co., LTD, and evaluated the safety and validity.Results The appearance integrity and stability of the main body of the biosafety cage changing workbench, the cleanliness of the working area, the velocity of the inflow airflow, the velocity of the descending airflow, the illuminance, the noise level, the airflow direction, the high-efficiency particulate air filter were at good condition, reaching the standards of level II biological safety cabinet. The airtightness of the contaminated goods recovery cabin and the sterilization chamber of utensil surface was good, but the size and quantity of the sterilized cage box need to be further improved.Conclusion The biosafety cage changing workbench developed by our team can be operated properly in whole. It can complete the replacement of individually ventilated cages in this workbench under negative pressure environment. Our workbench also can ensure the safety of operators, environment and experimental subjects.

    Performance Analysis of Class Ⅱ Biosafety Cabinetry in Veterinary-related Research Institutes
    Wenjie MA, Wei ZHANG, Wei WANG, Shibo SU, Huijuan GUO, He ZHANG, Zi WANG, Huairan LIU, Hongyan CHEN
    2022, 42(2):  111-116.  DOI: 10.12300/j.issn.1674-5817.2021.101
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    Objective To detect the performance and use of class II biosafety cabinets in laboratories of veterinary-related research institutes.Methods A total of 126 class II biological safety cabinetries being used by three veterinary-related reseatch institutes in Harbin were tested. The performance indicators were judged according to the test results and and analyze according to relevant standards, in order to understand the use of biological safety cabinets.Results The overall pass rate of 126 biosafety cabinetries was 17.5%. The total qualified rate were 100%, 100%, 100%, 100%, 49.2%, 92.9%, 68.3%, 22.2% and 100% ,respectively in terms of the appearance of integrity, cleanliness of work area, integrity of high efficiency filter for supply and exhaust air, flow pattern, the velocity of inflow, the velocity of the downdraft, noise, illumination and alarm and interlock system of class II biosafety cabinet.Conclusion The overall pass rate of biosafety cabinetries in some veterinary-related research institutes is low. Regular inspection, operation and maintenance management should be strengthened to eliminate biological safety hazard timely and prevent infection accidents of laboratory.

    Tracking and Monitoring on Application of Micro-electrolyzed Sterile Water System in Laboratory Animal Barrier Facilities
    Wei ZHANG, Bin TANG, Yong ZHAO, Zhini ZHANG, Xinyan DENG
    2022, 42(2):  117-121.  DOI: 10.12300/j.issn.1674-5817.2021.154
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    Objective To track and observe whether the long-term application of micro-electrolyzed sterile water system in laboratory animal barrier facilities can continuously maintain the terminal water aseptic.Methods From April 2018 to March 2022, the microbiological detection of the micro-electrolyzed sterile water system had been tracked and monitored once each quarter in the barrier facilities of producing and breeding SPF grade rats and mice in Laboratory Animal Center of Sun Yat-sen University. Totally 8 sampling points were set, including one sampling point before the intake of the microelectrolytized water outlet into the water duct, six sampling points at the terminal water outlet, and one sampling point for the water return of the water network. Samples were taken according to aseptic operation procedures, and microelectrolysis aseptic water was detected according to the national standards GB 14925—2010 and GB/T 14926.41—2001.Results From April 2018 to March 2022, a total of 17 tests were conducted. During the normal operation of the micro-electrolyzed sterile water system, the quality of micro-electrolyzed water at 8 sampling points was sterile, in line with the national standard.Conclusion Micro-electrolyzed sterile water system can maintain the sterility of the terminal water supply, and can be used in laboratory animal barrier facilities for a long time.

    Preliminary Exploration of Information Supervison System for Harmless Disposal of Laboratory Animal Carcasses and Related Wastes
    Yong LI, Liang ZHOU, Yong HE, Zhifeng YAN, Haijiang WANG
    2022, 42(2):  122-126.  DOI: 10.12300/j.issn.1674-5817.2021.075
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    At present, there is still a lack of standardized and institutionalized management of laboratory animal carcasses and related wastes in China, and effective supervision of their harmless disposal process, especially an effective whole-process biosafety supervision system. Using computer networks, global satellite positioning systems, wireless communications, geographic information systems and other multidisciplinary cutting-edge technologies, we preliminarily established an information management system for harmless disposal of laboratory animal carcasses and related wastes, which could be closely combined with biosafety supervision to realize dynamic tracking and monitoring in the whole process. The established system can strengthen the standardization and institutionalization management of experimental animal carcasses and related wastes, promote the safety control of medical waste, prevent the spread of diseases, protect the environment and ensure human health.

    Animal Models of Human Diseases
    Establishment and Comparison of Mouse Pancreatic Cancer Orthotopic Model for Minimally Invasive Ablation Therapy Study
    Lujing MAO, Nana ZHANG, Hao LIU, Aihua SHI, Ziyu ZHU, Yi LÜ
    2022, 42(2):  127-134.  DOI: 10.12300/j.issn.1674-5817.2021.094
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    Objective To explore the advantages and disadvantages of orthotopic mouse model of pancreatic cancer established by orthotopic injection of tumor cells and transplantation of tumor tissue block, in order to provide technical reference for the establishment of orthotopic mouse model of pancreatic cancer with the purpose of precision ablation treatment.Methods A total of twenty healthy aged 6-8 weeks male C57BL/6 mice were divided into four groups, with five mice per group. Direct injection of mouse pancreatic cancer Panc02 cell was used in group A, direct injection of mouse pancreatic cancer Panc02 cell (containing matrigel) was used in group B, tumor tissue block transplantation of mouse pancreatic cancer was used in group C, the tumor tissue block hydrogel bonding method was used in group D. The tumor formation rate, tumor size, the degree of abdominal organ adhesion, tumor metastasis were observed after surgery in each group of mice. Tumor histomorphology and expressions of Ki67 and α-smooth muscle actin (α-SMA) protein were detected by HE staining and immunohistochemistry, repectively.Results The tumor formation rates in groups A, B, C, and D were 60%, 80%, 100%, and 100% respectively. After modeling for 16 d, the tumor volume of mice in group A was (47.80±42.99) mm3, with poor homogeneity. The tumor volume of mice in groups B, C and D [(68.43±16.77) mm3, (105.86±17.25) mm3, (128.98±13.41) mm3 , respectively] was significantly larger than that in group A (P < 0.01), with better homogeneity. The severe abdominal adhesion rate in group A mice was 100% and 80% in group B, and no severe abdominal adhesions occurred in groups C and D. The metastasis rate was 100% in group A and 60% in group B, and no tumor metastases were found in mice in groups C and D. There was no difference in the tumor formation of the four groups of mice, and no significant difference in the expression of Ki67 and α-SMA proteins in the tumor tissues (P > 0.05).Conclusion The establishment of orthotopic pancreatic cancer in mice by tumor tissue transplantation and hydrogel adhesion has the advantages of high tumor formation rate, less metastasis, mild adhesion of abdominal cavity, no change in the biological characteristics of tumor cells, easy operation and popularization, and better modeling effect.

    Effects of Inagliptin on the Expression of Periosteal and TGF-β1 in Renal Fibrosis of Type 2 Diabetic Rats
    Huijie GONG, Guangming ZHANG
    2022, 42(2):  135-140.  DOI: 10.12300/j.issn.1674-5817.2021.093
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    Objective To analyze the effect of linagliptin on the expression of periostin and transforming growth factor-β1 (TGF-β1) in the process of renal tissue fibrosis in type 2 diabetic rats, and explore the anti-inflammatory and anti-fibrotic protective mechanism of linagliptin on diabetic nephropathy.Methods Totally twenty-four SD rats were injected with streptozotocin in intraperitoneal to replicate the type 2 diabetes rats, and another 12 SD rats were injected with the same dosage of citrate buffer as the control group. The type 2 diabetic rats were randomly divided into two groups: model group and treatment group. The rats in the treatment group were continuously given liggliptin (10 mg?kg-1?d-1) for 12 weeks, and the other two groups were given the same amount of normal saline. The pathological changes of renal were observed by HE staining and Masson staining. Immunohistochemistry and RT-PCR were used to detect the expressions of periostin and TGF-β1 protein and mRNA levels in kidney tissues of rats, respectively.Results HE staining showed that compared with the control group, there were a large number of inflammatory cells in the renal stroma in the model group, while there were less inflammatory cells in the treatment group. Messon staining showed that compared with the control group, there were more collagen fibers in renal interstitial in the model group, while the collagen fibers in the treatment group was not obvious. Immunohistochemistry and RT-PCR showed that compared with control group, the protein and mRNA expressions of periostin and TGF-β1 in rat renal in model group were significantly increased (P< 0.05), while they were decreased in treatment group (P< 0.05).Conclusion Inagliptin maybe reduce the protein expression of periostin by inhibiting the overexpression of TGF-β1 in the kidney of type 2 diabetic rats, and slow down the progression of renal fibrosis.

    Model of Diffuse Cerebral Thrombosis Induced by Arachidonic Acid in Rats
    Long NING, Hang SUN, Jin FENG, Qingting MENG, Qian YANG, Fangyan HE
    2022, 42(2):  141-145.  DOI: 10.12300/j.issn.1674-5817.2021.091
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    Objective Establishment and optimization of arachidonic acid-induced diffuse cerebral thrombosis model in rats.Methods Arachidonic acid was injected into the internal carotid artery to establish a diffuse cerebral thrombosis model in rats, while the sham operation group was set as the control. The model was evaluated by detecting the exudate content of Evans blue (EB) in brain tissue and the pathological damage degree of brain tissue by HE staining.Results Compared with the sham operation group, diffuse EB exudation was observed in the injured brain tissue of the model group (P < 0.01), and the exudation was relatively uniform. The HE staining of brain tissue on the side of the injury showed ischemic pathological damage with disorganized cell arrangement, wrinkled nuclei, enlarged cell gaps, increased vacuolated cells, and red striped tubular shape of each individual microvessel.Conclusion Arachidonic acid can successfully induce diffuse cerebral thrombosis model, and the optimized model has better stability and lower mortality.

    Progress of Pancreatic Stellate Cells in Research of Pancreatic Disease Model
    Xun WEI, Xia JIANG, Yuxuan ZHENG, Lewen ZHANG, Zhengyu ZHOU
    2022, 42(2):  146-151.  DOI: 10.12300/j.issn.1674-5817.2021.088
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    Common types of pancreatic diseases are pancreatitis, diabetes and pancreatic cancer. Clinical studies have found that pancreatic stellate cells (PSCs) in patients with pancreatic diseases, contributes to fibrosis, oxidative stress and inflammation, and lead to pancreatic diseases. This article reviewed the modeling methods of different pancreatic diseases, and the influence and application prospects of PSCs in pancreatic diseases, providing a new idea for the research and treatment of pancreatic diseases.

    Laboratory Animal Welfare and Ethics
    Improving Effect of Environmental Enrichment on Stress-related Indicators of British-shorthair Cats
    Hongting CHEN, Fei YANG, Ying HU
    2022, 42(2):  152-158.  DOI: 10.12300/j.issn.1674-5817.2021.130
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    Objective To investigate the effect of environmental enrichment on alleviating stress in laboratory British-shorthair cats.Methods A total of fourteen laboratory British-shorthair cats were randomly divided into welfare group (n = 7) and control group (n = 7) according to the presence of environmental enrichment interventions which lasted for 11 weeks. The hematological and blood biochemical, neuroendocrine and immunological parameters of each group were measured and compared to evaluate the stress level of different groups.Results The mean corpuscular volume in the welfare group was significantly lower than that in the control group (P < 0.05), the percentage of lymphocyte was extremely higher than that in the control group (P < 0.01), and the neutrophilic percentage was extremely lower than that in the control group (P < 0.01). In addition, the levels of carbamide, creatinine, globulin, total protein in the welfare group were significantly lower than those in the control group (P < 0.05); the adrenaline and dopamine indicators in the welfare group were significantly lower than those in the control group (P < 0.05), while interferon-γ (IFN-γ), interleukin (IL)-1β, IL-2, IL-4, IL-5, IFN-γ/IL-4 ratio, IL-2/IL-4 ratio, IFN-γ/IL-5 ratio and IL-2/IL-5 ratio of the welfare group were higher than those of control group; among them, IFN-γ, IL-2, IFN-γ/IL-5 ratio and IL-2/IL-5 ratio were significantly different (P < 0.05), and IL-5 difference was extremely significant (P < 0.01).Conclusion The environmental enrichment interventions can effectively improve the welfare level of laboratory cats and reduce the biological costs of stress.

    Application of Isolated Guinea Pig Ileum Method for Detection of Histamine Substances in Osteopeptide Injections
    Chang WU, Xiaojie LI, Yinnan WU, Yao WANG, Yuxin WANG, Weifeng ZONG, Changhong MENG, Yihong LU
    2022, 42(2):  159-165.  DOI: 10.12300/j.issn.1674-5817.2021.082
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    Objective To evaluate the detection of the histamine substances in osteopeptide injections by the isolated guinea pig ileum method, and to explore the feasibility of replacing cat depressor substances method.Methods According to the 2020 edition of Chinese Pharmacopoeia, the histamine substances in 7 batches of osteopeptide injections were detected from 5 enterprises by isolated guinea pig ileum method. The results were compared with the test method of cat depressor substances.Results By the isolated guinea pig ileum method, the histamine substances in 3 batches of preparations were negative, which was consistent with the results of the test method of cat depressor substances. The results of the other 4 batches of preparations were suspicious; while the examination results of depressor substances in cat were 3 batches suspicious and 1 batch positive. The overall consistency rate of the two methods was 85.7% (6/7).Conclusion The results of isolated guinea pig ileum method is almost the same as the test of cat depressor substances, which can be used for the screening and study of histamines in osteopeptide injections. It is suggested to add the inspection items of the test of depressor substances or histamines in the standard of osteopeptide injections.

    Nursing Practice and Discussion of Electrode Implantation into Cerebral Cortex of Rhesus Monkey
    Xibin XU, Qing ZHU, En ZHANG
    2022, 42(2):  166-170.  DOI: 10.12300/j.issn.1674-5817.2021.078
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    The use of electrode implantation technology to explore the electrical activity of population neurons is crucial for brain and cognitive science, brain-like research and brain-computer interface technology research. This article introduces the nursing points and significance of electrode implantation in rhesus monkey cerebral cortex before, during and after operation. Preoperative care includes preoperative preparation, preoperative medication, cranial fixation, skin preparation, etc. Intraoperative care includes physiological index monitoring and cerebral cortex implantation surgery care, while postoperative care includes observation and monitoring, debridement and nutritional care. Doing a good job in the operation of electrode implantation into the cerebral cortex of rhesus monkeys can not only improve the success rate of the operation, use as few experimental animals as possible to obtain the data required for the experiment, but also reduce the pain and injury caused by the operation to the animals, which is of great significance for brain and cognitive science research and animal welfare protection.