Table of Content

25 December 2016, Volume 36 Issue 6

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Immunogenicity of Coxsackievirus Group A Type 16 Strains in Mice

MENG Hua-qing, YANG Ting, XIE Tian-hong, LI Hua, YUE Lei, SONG Xia, ZHANG Ye, XIE Zhong-ping

2016, 36(6):  409-414.  DOI: 10.3969/j.issn.1674-5817.2016.06.001

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Objective To evaluate immunogenicity of Coxsackievirus group A type 16 strains (CA16) in mice and provide experimental data for further research. Methods The female BALB/c and ICR mice ageing 4, 6, 8 weeks were inoculated subcutaneously, intraperitoneally and intramuscularly with two different doses of CA16 live virus. Blood were respectively taken at 7, 21, 28 and 35 days after inoculation and serum neutralizing antibody titer was analyzed by micro-neutralization test in vitro and six cytokines were determined by ELISA. Results Seroconversion rates of the mice inoculated intramuscularly, intraperitoneally and subcutaneously were 83.33%, 40.00% and 0.00% at 28 days after the first inoculation respectively, and antibody geometric mean titers (GMT) were 1∶169, 1∶32 and 0. Seroconversion rates of the mice inoculated intramuscularly, intraperitoneally and subcutaneously were 100% at 7 days after the boost, and GMT were 1∶813 (4.8-fold), 1∶609 (19-fold) and 1∶32; Seroconversion rate was 100%, which was significantly higher than that of 28 and 21 days at the first inoculation. Neutralizing antibody titersand seroconversion rates of the Balb/c were significantly higher than that of ICR, but effect of the age on the neutralizing antibody titers and seroconversion rates was not significant (P>0.05). The dose has significantly effect on the neutralizing antibody titer and seroconversion rates (P<0.05). In high dose group, the neutralizing antibody titer and seroconversion rates was significantly higher than low dose. interleukin(IL)-10 and tumor necrosis factor (TNF)-a have a significant increase at 7 days after the boost, compared to the 7 days after the first inoculation. Conclusion BALB/c and ICR mice can be used as animal models of CA16 live virus immunogenicity evaluation, and the most sensitive models was 4-6 week old BALB/c mice.

Survey on Intestinal Amoeba in Diarrhea Tree Shrews

SONG Qing-kai, MIAO Yu-run, YIN Bo-wen, CHEN Ling-xia, DAI Jie-jie, SUN Xiao-mei

2016, 36(6):  415-418.  DOI: 10.3969/j.issn.1674-5817.2016.06.002

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Objective To investigate the infection status of intestinal Amoeba in diarrhea tree shrews from the center of germplasm resources, to find out the cause of diarrhea, and to provide useful reference and support for prevention and treatment of parasitic infection in tree shrews. Methods Totally 78 samples were collected from the fecal and blood of diarrhea tree shrews, and then were detected by the normal saline direct smear method and iodine solution staining, meanwhile stool antigen and IgG antibody in serum were detected by ELISA of Amoeba. Results The results showed that the positive infection rate were 42.22%(19/45) of wild tree shrews, 33.33%(11/33) of breeding of tree shrews by direct smear and iodine staining method. The infectious rates of E. histolytica were 64.44%(29/45) and 42.42%(14/33)respectively when detected the stool antigen by ELISA. The positive rate of Amoeba IgG were 66.67%(30/45) and 63.63%(21/33) by ELISA detection. Conclusions Amoeba infection rate is rather high in tree shrews, and it may be one of the pathogens of diarrhea tree shrews. In routine breeding should be strengthened in the process of testing control the spread of the parasite.

Molecular Subtyping Study on Pseudomonas aeruginosa in Rats and Mice with Pulsed Field Gel Electrophoresis

FENG Jie, XIE Jian-yun, WEI Xiao-feng, HU Jian-hua, GAO Cheng

2016, 36(6):  419-422.  DOI: 10.3969/j.issn.1674-5817.2016.06.003

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Objective To investigate the correlation between Pseudomonas aeruginosa epidemic strains and host in recent years, Pseudomonas aeruginosa in rat and mice from several faciliies of Shanghai and Jiangsu province were isolated and detected by pulsed field gel electrophoresis (PFGE). Methods The bacteria were isolated from the contents of ileocecal junction under aseptic conditions. The isolated strains were identified by morphological observation and physicochemical test. Extracted genomic DNA of Pseudomonas aeruginosa isolates were detected by PFGE after digested by Xba I restriction enzyme. Results A total of 67 strains were isolated from rats and mice. The genotype of 67 strains was divided into 39 patterns and 14 clusters (A-N). The dominant cluster is I, H, B and G, the rate of these cluster were 17.9%(12/67), 14.9%(10/67), 13.4%(9/67) and 11.9%(8/67) respectively. The isolates from the same source mostly present different gene cluster. Conclusion A great deal of gene polymorphism could be found in all the 67 strains which are diffusively distributed. Obvious prevalent feature could not be found.

Establishment and Evaluation of Rat Model of Neurogenic Bladder after Spinal Cord Transection Injury

YUAN Xiao-hong, HE Feng, JIANG Ze-hui, ZHAO He, YE Chao, WU Shao-ming, YU Hai-chuan, LI Chun-gen

2016, 36(6):  423-427.  DOI: 10.3969/j.issn.1674-5817.2016.06.004

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Objective To establish the ideal animal model of neurogenic bladder (NB) after spinal cord injury (SCI) and evaluate and the state of the bladder. Methods Sixteen female SD rats were divided into control group and the experimental group (T9 spinal cord transection injury model), according to the record of the daily manual urination output to evaluate the NB recovery after SCI. After two weeks the simultaneous recording of the intravesical pressure were used to detect, compare and assess the state of bladders in two groups. Results The urine output of experimental rats at first day, 5^th day, 10^th day, 13^th day, 14^th day after operation were significantly higher than thoes of control group rats (P<0.05), and the urine output of experimental rats at 16^th day after operation was no statistically significant difference with control group (P>0.05). During a week postoperation, the daily manual urination output increased and reached maximum. One week later postoperation, the daily manual urination output was on the gradually decrease and stable for two weeks. The values of the parameters of the maximum bladder pressure, the basis bladder pressure, voiding threshold, bladder capacity, voiding efficiency in control group and experimental group respectively were 26.60±4.31 mmH₂O and 21.66±2.56 mmH₂O, 11.66±2.33 mmH₂O and 14.72±2.56 mmH₂O, 20.46±0.52 mmH₂O and 16.99±0.81 mmH₂O, 1.36±1.58 min and 2.02±0.36 min, 0.82±0.15 mL and 2.20±0.24 mL, 92.67%±1.97% and 25.33%±4.46%, the differences were statistically significant (P<0.05). Conclusion The NB rat model of T9 level spinal cord transection injury with a strong maneuverability and easy to quantify, repeatable. With the correct postoperative nursing care, there were fewer complications and low mortality. The recording of daily manual urination output and the intravesical pressure can be the good ways to detect the spinal cord recovery and evaluate the status of the bladder.

Establishment of Brain Injury Model in Mouse by Intracranial Injection of Anhydrous Ethanol

HU Fan, DAI You-Jin, HOU Dao-Rong

2016, 36(6):  428-432.  DOI: 10.3969/j.issn.1674-5817.2016.06.005

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Objective To establish a stable mouse model of brain injury by ethanol intracranial injection. Methods Sixty ICR mice were randomly divided into 6 groups: sham group (n=10), 5 mL group (n=10), 10 mL group (n=10), 15 mL group (n=10), 20 mL group (n=10) and 25 mL group (n=10). The mice were intranscranially injected with different doses of ethanol. After 7 days, pathological examination was performed and the level of malondialdehyde (MDA), superoxide dismutase (SOD) and relevant genes expression were detected. Results After model made, the ethanol injection mice showed walk and balance function obstacle at early time, and returned to normal later, but feed intake and body weight significantly decreased. With HE staining, the nerve cell necrosis and surrounding tissue edema, neurons and glial cells intercellular space expansion, microvascular clearance outside enlarge and vascular diapedesis, the nerve cells and glial cells cavity lesions were detected. The activity of SOD were decreased and content of MDA were increased. The cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) and synaptosomal-associated protein-25 (SNAP-25) mRNA expression was significantly increased in experimental group compared with control group, detected by Q-PCR. Conclusion The mouse model of brain injury made by intracranial injection with ethanol have the advantages of simple, phenotypic uniformity, good repeatability, high success rate compared to traditional brain injury model making.

Comparative Study on Iron Deficiency Anemia Rats Model by Three Different Formula Feeds

DING Guang-yu, PAN Mian-li, HU Han, LIU Yi, SHEN Long-hai

2016, 36(6):  433-436.  DOI: 10.3969/j.issn.1674-5817.2016.06.006

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Objective To explore the effect on iron deficiency anemia rat model induced by 3 different formula feeds. Method Wistar rats at age of 3~4 weeks were randomly divided into 4 groups: group A, group B, group C and control group. The groups A, B, C were fed with 3 different formula feeds respectively, and control group was fed with normal feed. Experiment lasts for 35 days, the body weight and food intake of rats in each group were measured every week. Meanwhile, behavior and appearance sign was noted during the experiment. Rat抯 blood was collected and tested after 35 days, including hemoglobin, hematocrit, etc. Results Hemoglobin(Hb) concentration of group A decreased to 90 g/L and morphological features met the requirments. Hb in group B did not decrease; Hb in group C decreased a little but no less than 90 g/L. Conclusion The formular A feed can establish iron deficiency anemia model successfully, formula B and C are not applicable for the model building.

Nephron Sparing Surgery by Regional Occlusion in Dog

FU Dian, CHENG Wen, TIAN Feng, LI Ping, ZHANG Zheng-yu, GE Jing-ping, GAO Jian-ping

2016, 36(6):  437-440.  DOI: 10.3969/j.issn.1674-5817.2016.06.007

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Objective To investigate the nephron sparing surgery by regional occlusion with dogs. Methods By using the regional blocking renal blood tool developed by the authors, 5 healthy male Beagle dogs underwent nephron sparing surgery (NSS). While 3 healthy male Beagle dogs underwent NSS by blocking the renal artery. Results All the 8 Beagle dogs successfully underwent the surgery and had excellent recovery. All the outcome measures between two groups had no statistically significant differences. Conclusion Nephron sparing surgery by regional occlusion using our self-made tools is safe and effective. This method might be better able to protect renal function.

Investigation on Biological Characteristics of Blood for Four Breeds of Minipigs

YANG Li-chang, ZHOU Wen-bing, DING Jun, ZHOU Xin-chu, ZHANG Jian-ping, XIE Dong, YANG Wei-min

2016, 36(6):  441-446.  DOI: 10.3969/j.issn.1674-5817.2016.06.008

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Objective To analyze and compare of the biological characteristics of the blood of four breeds of minipigs. Method The four breeds of minipigs including Bama minipigs, Guizhou minipigs, Banna minipigs and Luchuan minipigs at the age of 3 to 6 months reared in Shanghai area were selected to determine the hematology, serum biochemistry and blood coagulation values with 40 (half males and half females) in each breed. All data were analyzed by one-way ANOVA method and then compared. Result Among the four breeds of minipigs, 14 parameters of hematology, 12 parameters of serum biochemistry and 4 parameters of blood coagulation were significant different (P<0.05). Compared the results between males and females, 6 hematologic parameters, 4 serum biochemistry parameters and 3 blood coagulation parameters in Bama minipig were significantly different (P<0.05), a few parameters or no parameters of blood in Guizhou minipigs, Banna minipigs and Luchuan minipigs were significant different. There are many parameters existing significant different between same gender among four minipigs breeds. Conclusion The parameters of hematology, serum biochemistry and blood coagulation of Bama minipigs, Guizhou minipigs, Banna minipigs and Luchuan minipigs provide the abundant biological data support when applying and selecting minipig for medical science model research and pharmaceutical research.

Choice of Humane Endpoint of Laboratory Animals in Typical Developed Countries

TAO Ling-yun, ZHOU Jie, NI Li-ju, CHEN Yi-fei, GAO Cheng

2016, 36(6):  451-454.  DOI: 10.3969/j.issn.1674-5817.2016.06.010

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As the important experimental materials for life science, laboratory animals will be faced with the choice of humane endpoint after achieved the scientific goals and objectives. Laboratory animal welfare laws and regulations are relatively perfect in typical developed countries. Studying the choice and practice of humane endpoint in the laws and regulations of experimental animals in typical developed countries has the enlightenment function to laboratory animal welfare legislation in China.

Role of Laboratory Animal Veterinarian in Hospital

XU Hu-feng, SUN Chen-yang

2016, 36(6):  455-460.  DOI: 10.3969/j.issn.1674-5817.2016.06.011

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Laboratory animal veterinarian is not equal to traditional animal veterinarian. Besides of the health care of laboratory animals, the work of laboratory animal physician covers euthanasia and clearance of dead body and waste. Because of the great duty of laboratory animal veterinarian, they play a central role in animal laboratories. According to the detailed work of laboratory animal veterinarian executed in Beijing Friendship Hospital, this article elaborates the practice and duty of laboratory animal veterinarian to offer information to related people working in the same field.

Design and Application of Laboratory Animal Experiment Management System

XUE Qi, ZHANG Ying-hui, WU Si-jie, REN Xiao-xia, YAO Wen-sheng

2016, 36(6):  461-465.  DOI: 10.3969/j.issn.1674-5817.2016.06.012

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This paper briefly introduced the design objective, the basic function and the characteristics of laboratory animal experiment management system in China Institute of Veterinary Drugs Control. The system has realized the information operations of the ethical review of animal experiments, the approval of biological safety, the application and registration of animal experiments. The system can also realize no paper working, solve the problem of the work in different locations, and improve the analytical ability of animal experiments data statistics. This paper investigates the problem and development of the system simultaneously. It may provide a reference of animal experiment management system for other research institutions.

Preparation of Pre-eclampsia Animal Model

LI Dong-dong, ZHU Min, PANG Qin-xia, Ding Pei-fang

2016, 36(6):  466-472.  DOI: 10.3969/j.issn.1674-5817.2016.06.013

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Pre-eclampsia (PE) is a common obstetrical and gynecological disease with the hypertension, proteinuria and edema as the main clinical symptom, and accompanying with multiple organ damages. The disease is a serious threat to maternal and child safety and the leading cause of maternal and perinatal morbidity. However, the etiology and pathogenesis of PE is not clarified yet. Using animal models of PE to research human diseases is an important mean to clarify the pathogenesis of the disease or drug efficacy. There are four methods of PE animal modeling: environmental stimulation, chemical drug interference, surgical intervention and gene intervention. Rats and mice are mainly selected to prepare animal models of the PE. In addition, rabbits, dogs, rhesus monkeys, baboons and sheep are also commonly used for preparing the animal models by surgical intervention. In this paper, the preparation methods of PE animal models were reviewed, and the evaluation standards and characteristics of these animal models were introduced.



Research Progress on Toxic Milk Mouse

ZHANG Wei, TANG Bin, LI Fu-rong

2016, 36(6):  473-478.  DOI: 10.3969/j.issn.1674-5817.2016.06.014

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Toxic milk mouse is mutant a strain that produced by DL mice at F68. The main pathological shows are low pigmentation, growth retardation, tremor and movement disturbance. The common ground with Wilson disease are autosomal recessive inheritance, and large quantities of copper deposit in adult mouse that lead the levels of serum copper and serum ceruloplasmin protein declined. This paper reviews the researches on Toxic milk from several aspects, including pathophysiology, disease mechanism, diagnostics and therapy.