Laboratory Animal Science

Caesarian Purification of Chinese Hamster

2020, 37(04): 1.

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Objective In order to make the quality control level of microbiology and parasitology of Chinese hamsters reach the clean standard through biological purification. Method First of all, the offspring of ordinary Chinese hamsters were delivered by cesarean section, and then the milk of clean grade ICR mice was substituted. At the same time, the changes of body weight and survival of 30-day-old offspring were recorded, and the microbiological and parasitological quality of weaned mice were tested. Result In this experiment, 24 nests of ordinary Chinese hamsters were selected for cesarean section, and a total of 109 new born rats were obtained, of which 5 were dead, and 56 survived, with a success rate of 53. 85%. After weaning, all the hamsters reached the clean national standard. Conclusion This study carried out biological purification of Chinese hamsters, further improved the quality standardization of Chinese hamsters, and provided quality guarantee for the production and application of Chinese hamsters.

Detection Results and MLST Analysis of Staphylococcus aureus in Laboratory Rats from Beijing Area

2020, 37(04): 5.

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Objective To understand the carrying status and molecular characteristics of Staphylococcus aureus in laboratory rats from Beijing area, provide basic information for the prevention and control of Staphylococcus aureus in laboratory rats. Method According to the detection method for Staphylococcus aureus in laboratory animal ( GB / T 14926. 14—2001) , the laboratory rats from Beijing area from 2008 to 2017 were tested, and the detection status was classified by the year and season. Then, the main strains were typed and analyzed by MLST assay. Result From 2008 to 2017, 165 strains of Staphylococcus aureus were isolated in laboratory rats from Beijing, with a positive rate of 8. 44%. In 2013, the positive peak of Staphylococcus aureus (39 strains, 18. 48%) was reached, and the positive rate wasn’ t significantly different between seasons. MLST classification divided 148 strains of Staphylococcus aureus into 7 types ( ST8, ST3, ST1, ST6, ST2871, ST88 and ST641) , of which ST1 accounted for the largest proportion (51. 35%) , followed by ST2871 (30. 41%) . The remaining ST types accounted for less than 10%; further analysis found that it has significant spatial and temporal distribution characteristics, and there is evolutionary correlation. Conclusion This study provides basic information for the prevention and control of Staphylococcusaureus in laboratory rats.

Specific Desensitization Therapy Forallergic Rhinitis Caused by Willow Pollen

2020, 37(04): 10.

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Objective Establish mouse models of willow pollen allergic rhinitis and immune tolerance treatment of the mouse models. Method Pollen protein was extracted and mouse models of allergic rhinitis was established by intraperitoneal injection of pollen protein and nasal stimulation. By subcutaneous injection of increasing dosage of allergen into mice, the behavioral symptoms of mice were observed, the morphological changes of nasal mucosa were compared, and the content of specific sIgE in serum was detected, so as to study the therapeutic effect of specific desensitization therapy on allergic rhinitis. Result Before treatment, allergic symptoms such as scratching, sneezing and runny nose appeared in mice. Eosinophilsin nasal mucosa were significantly more than those in control group, and the content of specific sIgE in serum was higher than that before stimulation. These results indicate that the allergic rhinitis mouse models induced by willow pollen has been successfully established. After treatment, allergic symptoms of mice gradually decreased to disappear, eosinophils in nasal mucosa gradually decreased, and specific sIgE content in serum decreased. Conclusion Willow pollen can establish allergic rhinitismodelsin mice, and specific desensitization method could treat allergic rhinitis in mice.

Establishment of a Fluorescent Quantitative PCR Detection Method for Sendai Virus and its Preliminary Application

2020, 37(04): 21.

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Objective To establish a fluorescent quantitative PCR ( Q-PCR) method for the Sendai virus ( SeV) . Method A fluorescent quantitative PCR ( Q-PCR) method for SeV was developed based on a pair of primers that in accordance with the published sequence of SeV L gene. The specificity, sensitivity, repeatability and stability of the method were verified. The method was used to detect 40 SPF mice,58 naked mole rats, 4 Spermophilus dauricus and 5 clean mice. Result The assay could specifically detect SeV and had good sensitivity, the minimum detectable amount of Sendai virus was 10 copies / μL. The 40 SPF mice and 58 naked mole rats were negative by this Q-PCR assay,4 Spermophilus dauricus and 5 clean mice were all positive. Conclusion The developed Q-PCR method is good in linearity, specificity, sensitivity, and can be used for the rapid quantitative detection of SeV samples. It provides technical reference for the monitoring of SeV and improvement of related standards.

Observation of Angiogenesis of Two Kinds of Subcutan Transplanted Tumors in Mice by Ultrasound

2020, 37(04): 27.

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Objective To observe the angiogenesis of human bladder cancer ( BT-B ) and human liver cancer ( Huh7) subcutaneously transplanted tumors by high frequency ultrasound in small animals. To evaluate the value of contrast-enhanced ultrasound in tumor angiogenesis. Method Two subcutaneous transplantation tumor models of human BT-B bladder cancer and human Huh7 liver cancer were established by subcutaneous injection. The number and size of small blood vessels were observed by confocal laser probe in vivo. The expression of CD31, a tumor vessel marker, was observed by immunohistochemistry and the microvascular density was calculated. Result Both transplanted tumors showed good growth patterns. B-mode ultrasound showed that both Huh7 and BT-B subcutaneous tumors were heterogeneous hypoechoic nodules. Color doppler mode and spectral doppler mode showed that the blood flow signal and velocity of BT-B subcutaneous tumor were higher than that of Huh7 subcutaneous tumor. Contrast-enhanced ultrasound showed that the subcutaneous tumor of Huh7 presented diffuse and uneven enhancement, and the peripheral ring or semi-ring enhancement of the subcutaneous tumor of BT-B presented enhancement toward the center of the lesion, which was consistent with the probe in vivo laser confocal result and pathological result . Peak intensity of CEUS parameters ( PI) was positively correlated with MVD ( r = 0. 844, P < 0. 05) . Conclusion CEUS can better display the distribution of tumor blood vessels, and PI is positively correlated with pathological MVD, suggesting the proliferation of tumor blood vessels, which can be used as a reliable method for noninvasive evaluation of tumor angiogenesis.

Pathological Diagnosis and Analysis of Hair Follicle Tumors in Dogs

2020, 37(04): 33.

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Objective Skin tumor is one of the most common tumors in veterinary medicine, and is probably the most common histopathological specimen sent to the diagnostic laboratory, and is often submitted for histopathological evaluation. Method From 2018 to 2019, a total of 39 cases of tumors originated from skin hair follicles were examined in dogs from different units. Pathological diagnosis and analysis were carried out on the 39 cases of tumors, and the tumor types, age, gender, benign and malignant, dog breeds and other aspects were analyzed in order to explore the rule and pathological characteristics of tumors originated from skin hair follicles in dogs. Result The highest incidence of hair follicle tumors was trichoblastoma (73%) , and the most common type in dogs was the ribbon type. In terms of benign and malignant tumors, the incidence of benign tumors is very high, accounting for 97%. The incidence of hair follicle-derived tumors in dogs tends to increase with age, and the incidence of these tumors is higher in older dogs. The occurrence of tumors originating from skin follicles did not show significant gender preference. Conclusion It can be used as a reference for the diagnosis and differential diagnosis of tumors originated from skin follicles.

Safety and Immunogenicity of Split Seasonal Influenza Vaccine in BALB / c Mice

2020, 37(04): 44.

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Objective To evaluate the safety and immunogenicity of split seasonal influenza vaccine in BALB / c mice. Method The toxicity test was performed according to the requirements in Chinese Pharmacopoeia( Volume Ⅲ ,2015 edition) . For immunogenicity, BALB / c mice were divided into two groups randomly. The mice in test group were immunized with split vaccine at a dosage of 0. 1 mL intramuscularly, and those in negative control group with PBS. The mice were monitored daily for their activities, diet and weight up to day 49 post- immunization. Sera were harvested from the mice on day 0, 14, 28, 35 and 49 for seroconversion confirmation using hemagglutination inhibition ( HI ) assays. Result Our toxicity test results were consistent with the requirements in Chinese Pharmacopoeia( Volume Ⅲ , 2015 edition) ,and the vaccine was safe and had no effect on the growth of mice. The HI antibody of 100% antisera raised against A( H1N1) pam09 and A( H3N2) were positive after immunization 28 days, and 70% antisera raised against B ( Victoria lineage ) were positive after immunization 35 days. The concentration of the antibodies reached the peak after immunization 35 days. Our result indicated that the tested vaccine could elicited antibodies against A ( H1N1 ) pam09, A ( H3N2 ) and B ( Victoria lineage ) by first immunization. Conclusion The split seasonal influenza vaccine was safe and efficacious in BALB / c mice.

Protectin DX Protects Lung Injury-related Inflammatory Factors and Oxidative Damage in Aged Rats

2020, 37(04): 49.

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Objective The role of Protectin DX ( PDX) in the regulation and protection of inflammatory mediators and oxidative factors related to lung injury after lobectomy in elderly rats was explored. Method Thirty healthy elderly male SD rats (10 months old) were randomly divided into the sham operation group, the surgery group and the PDX treatment group ( n = 10) . In the sham operation group and the operation group, 300 μL of physiological saline was injected intraperitoneally 1 hour after the modeling, and in the PDX treatment group, 300 μL of PDX (500 ng) was intraperitoneally injected 1 hour after the modeling. All rats were sacrificed by over-anaesthesia 24 h after modeling, and specimens were collected. The pathological changes of lung samples in each group were observed with HE staining, and the dry-wet mass ratio ( W / D) of lung samples in each group was evaluated. The protein concentration in the bronchoalveolar lavage fluid ( BALF ) was detected; the inflammation factors in the BALF was detected; the content of MDA, SOS, SOD and the expression of HO-1, nuclear factor ( NF) -κB protein in lung tissue were detected. Result The lung tissue structure of the sham operation group was clear, and no inflammatory cell infiltration was seen; the lung tissue structure of the elderly rats in the operation group was disordered, accompanied by a large number of inflammatory cell infiltration. Compared with the sham operation group, the lung injury score and lung tissue W / D of the elderly rats in the operation group were significantly increased ( P < 0. 01) . The protein concentration and the contents of IL-1β, IL-6 and TNF-α in alveolar lavage fluid were significantly increased ( P<0. 01) , and the levels of MDA, ROS and NF-κB protein in lung tissues were significantly increased, SOD activity and HO- 1 protein expression was significantly reduced ( P<0. 05) . Compared with the surgery group, the lung injury score and lung tissue W / D of aged rats in the PDX treatment group were significantly reduced ( P<0. 01) . The protein concentration and the contents of IL-1β, IL-6 and TNF-α in alveolar lavage fluid were significantly decreased ( P<0. 01) , the MDA, ROS content and NF-κB protein expression in lung tissue were significantly reduced, SOD activity and HO-1 Protein expression was significantly increased ( P < 0. 05) . Conclusion PDX could significantly reduce the lung injury after lobectomy in elderly rats. This effect may be related to the regulation of HO-1 and NF-κB protein expression in the lung tissues, thereby inhibiting the oxidative stress and inflammation in the lungs.

ElasticSearch Technology in Animal Models

2020, 37(04): 55.

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Animal models are the important support tools of tumor research. In this study, the scalable big data retrieval technology originally used to integrate literature resources in animal models of tumor disease. This paper discusses a technology solution for integrate literature resources in animal models of tumor disease based on ElasticSearch. The system has four parts, including web crawler、 the Foundation Database、 ElasticSearch and Subscriber-Database. The result demonstrate that the developed system is superior to the system based on RDBMS. Having the characters of flexibility, briskness and efficient, the technology facilitate the further development of intelligent mining.

Case Analysis of Spontaneous Lipoma in Rhesus Monkeys

2020, 37(04): 59.

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A male rhesus monkey was found to have a bulging mass in the abdomen of the artificial monkey farm. Feeding, drinking water and defecation were normal. The palpation was palpated, the mass of the tumor was intact, the boundary of the mass was clear, the texture was hard, the parenchyma was strong, the irregular shape was about 10 cm in diameter, and there was no obvious pain in palpation. The tissue was inserted into the tumor with a needle to extract no liquid contents. The shape of the tissue in the mass was detected by B-ultrasound, the boundary was clear, the echo of the capsule and a little blood flow signal were cellulose-like changes; the body temperature was normal; the blood physiological and biochemical indicators were detected, and the result were compared with normal monkeys: ALP increased, TP, ALB, TC, RBC, HCT were reduced; a lumpectomy was performed, and a lump tissue was taken for microscopic examination to confirm the lipoma. The rhesus monkey has a good prognosis after postoperative care.

Policy Support and Consideration about the Development of Laboratory Animal Science and Technology in China

2020, 37(04): 64.

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Laboratory animals are the basic materials and supporting condition of life science research, important strategic scientific and technological resources, as well as an important part of the national science and technology platform construction. The national science and technology innovation plan of the 13th five year plan puts forward: “ develop the cultivation of new resources and varieties of laboratory animals, accelerate the creation and application of animal models of anthropogenic and complex diseases, and add a number of new varieties and new strains, with the total amount of resources close to the level of developed countries;. . . ” . Therefore, this paper combs the relevant policies of the development of laboratory animal science and technology in China from the aspects of laws and regulations, standard system, management system, etc. , and discusses the guiding role of these policies in the construction and sharing of laboratory animal resources in China, as well as the thinking and suggestions on improving relevant policies and regulations and implementing management measures.

Current Status and Prospect of Animal Experimental Study on the Treatment of Third Lumbar Transverse Process Syndrome with Acupotomy

2020, 37(04): 69.

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Objective To understand the main effect mechanism of acupotomology on the third lumbar transverse process syndrome ( TCS) by reviewing the literature on the basic research of TCS in recent years. Method Based on the current status of basic research on the animal model of third lumbar transverse process syndrome by acupotomy, this paper summarizes the animal experimental research on the treatment of third lumbar transverse process syndrome by acupotomy from the aspects of modeling method and different effect mechanisms. Result By comparing the experimental result , the therapeutic effect of acupotomy on the third lumbar transverse process syndrome was confirmed to some extent, which also provided reference and basis for clinical research and application.

Molecular Mechanism of Intestinal Flora Regulating Lipid Metabolism

2020, 37(04): 74.

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man intestinal flora genomes, also known as “ human second genomes,” play an important regulatory role in the decomposition and synthesis of dietary fats in the body. The disorder of the intestinal flora in the body can cause the occurrence of metabolic syndrome such as obesity and diabetes. In this paper, the progress of the study on intestinal flora providing energy, reducing fat and improving health by regulating enzymes and regulatory factors that play an important role in lipid metabolism is reviewed

Swine Models of Diabetes Mellitus

2020, 37(04): 78.

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Pigs are ideal animals for diabetes mellitus ( DM) research for its similarities with human beings on metabolism. The article reviewed the swine models of diabetes mellitus, which focuses on diagnosis of swine diabetes mellitus, type 1 DM, type 2 DM, genetic modified swine for DM and the nature strains / stocks susceptible to DM.

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