Laboratory Animal and Comparative Medicine

Establishment of Chronic Pulmonary Hypertension Model in Tibet Minipig

LIU Rong, HOU Meng, LIAO Hai-xing, ZHANG Xin-feng, ZHAO Jin, LIU Bao-hua, QIAN Yuan-xin, WU Qing-hong, MO Zi-yao, LIAO Dong-jiang, LI Hong-tao, GU Wei-wang

2015, 35(6): 431-435. DOI: 10.3969/j.issn.1674-5817.2015.06.001

Asbtract ( 282 )

PDF (1597KB) ( 454 )

References | Related Articles | Metrics

Objectives To establish chronic pulmonary hypertension (PH) model with monocrotaline (MCT) in young Tibet minipigs. Methods Twelve Tibet minipigs (weighing 5.0~6.0 kg, aged 2.0 months) were randomly divided into two groups and treated with MCT (12.0 mg/kg) or placebo (75% ethanol). After six weeks, the pulmonary arterial pressures of the minipigs were measured by performing right heart catheterization (RHC). The pathologic changes of small pulmonary arteries were observed. Index of right ventricular hypertrophy was also compared between two groups. Results The mean pulmonary arterial pressure by RHC was obviously increased in the MCT group than that in control group (24.62±1.38 mmHg vs 15.10±0.76 mmHg, P<0.01). The pathologic observations showed that the pulmonary vascular remodeling and the lumen disappeared in small pulmonary arteries. There was also progressive intimal fibrosis accompany with hyalinization, the lumen was occlusive, twisty and distorted with plexiform lesions. Indexes of right ventricular hypertrophy of MCT group were significantly higher (0.36±0.16 vs 0.28±0.13, P<0.05). Conclusions An animal model of chronic PH was established successfully in young minipigs at six weeks after MCT was intraperitoneally injected. The features of the chronic PH model of minipigs are similar with the primary PH parameters.

Expression of Pulmonary Surfactant-associated Protein A and CT Imaging in Lung Re-expansion Minipig after Atelectasis

TU Jun-wei, WANG Sai-bin, SHENG Yi-jun, CHEN Hui-jun, TIAN Jiang-hua, SHENG Lin, YING Ming-liang, Hu Bin, HE Zhong-ping

2015, 35(6): 436-514. DOI: 10.3969/j.issn.1674-5817.2015.06.002

Asbtract ( 263 )

PDF (1432KB) ( 456 )

References | Related Articles | Metrics

Objectives To investigate the expression of Pulmonary surfactant-associated protein A(SPA) and the CT imaging in lung re-expansion after atelectasis. Methods The minipig model of lung re-expansion after atelectasis was established. Male China Experimental Miniature Pig-I were randomly divided into three-days atelectasis group (3 d), one-week atelectasis group (1 week) and two-weeks atelectasis group (2 weeks), respectively (n=6). Pigs were euthanized after two weeks of lung re-expansion. The expression of SPA of experimental lung tissue was detected by immunohistochemistry. At specified time point, CT value and imaging changes were measured. Results The expression of SPA was lower in 2 weeks group than that in the other two groups (P<0.01). The CT value was lower in 3 d group than that in the other two groups (P<0.01), and the degree of lung re-expansion was the highest level in 3 d group among three groups. Conclusions The SPA expression of experimental lung tissue was decreased with atelectasis prolonged. The CT imaging indicated that improvement of lung re-expansion was dependent on the time of the atelectasis and the preferable opportunity of atelectasis cure was within one week.

Effects of High-fat Diet and Treadmill Exercise on AMP-activated Protein Kinase (AMPK)/Acetyl-CoA Carboxylase (ACC) Signaling Pathway and Fatty Acid Translocase CD36 Protein Content in Rat Gastrocnemius Muscle

ZHANG Yun-li, LOU Shu-jie

2015, 35(6): 441-447. DOI: 10.3969/j.issn.1674-5817.2015.06.003

Asbtract ( 205 )

PDF (917KB) ( 559 )

References | Related Articles | Metrics

Objectives To investigate the effects of high-fat diet and 8 weeks aerobic endurance exercise (treadmill exercise) on AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) signaling pathway and fatty acid translocase CD36 protein content in rat gastrocnemius muscles. Methods A rat model of nutritional obesity was established by feeding rats a high-fat diet, and then the obese rats were randomly divided into obese control group (OC group) and obese exercise group (OE group); In addition, a normal diet control group (NC group) and a normal diet exercise group (NE group) were set up. The exercise groups (OE and NE groups) were arranged by aerobic endurance exercise for 8 weeks. After the experiment, the levels of AMPKa, p-AMPKa, ACC, p-ACC and membrane protein CD36 in all groups were detected by Western blotting in gastrocnemius muscles. Results 1) Compared with the NC group, p-AMPKa protein levels had a significant increase in the NE group (P<0.01). p-AMPKa protein levels in the OE group were significantly higher than that in the OC group (P<0.01). p-AMPKa protein levels in the OC group were significantly lower than that in the NC group (P<0.01). 2) Compared with the NC group, p-ACC protein level in the OC group were significantly decreased (P<0.01). p-ACC protein levels had a significant increase in the OE group as compared with the OC group (P<0.01). 3) Compared between the NE group and NC group, the OE group and OC group, and the OC group and NC group, the content of membrane protein CD36 showed no significant change (P>0.05). Conclusion 1) The treadmill exercise could improve AMPK/ACC signaling pathway. 2) The impacts of exercise on p-ACC levels were different in rats with different body mass. 3) The membrane protein CD36 content in the rat gastrocnemius muscles did not significantly change whether AMPK/ACC signaling pathway was activated or inhibited.

Establishment of Preeclampsia Model in Rat and Detection on Related Indicators

ZHOU Jing, YAN Guo-feng, LUO Zhang-yuan, CHEN Dong-bao, CHEN Xue-jin, LI Yao

2015, 35(6): 448-452. DOI: 10.3969/j.issn.1674-5817.2015.06.004

Asbtract ( 330 )

PDF (1624KB) ( 361 )

References | Related Articles | Metrics

Objectives To establish preeclampsia SD rat model and monitor the alterations of related indicators in conscious rats. Methods Sixteen SD rats were divided into 4 groups randomly. Telemetrical modeling group A, which was implanted radiotelemetry and built preeclampsia model. Group B was modeling control group, which only built preeclampsia model without radiotelemetry implantation. Group C was implanted radiotelemetry and operated sham-operation of preeclampsia modeling. Group D was only operated sham-operation of preeclampsia modeling. Blood pressure of rats in groups A and C were monitored continuously after radiotelemetry implantation. Urine samples were collected from each group at pregnant day 13.5 and day 20.5 for urine TP and creatinine test. Pregnant rats were euthanieed at day 21.5 of pregnancy, and then the liver, kidney and placenta tissues were collected for HE staining. Results Mean arterial pressure, systolic pressure and diastolic pressure of group A were significant higher than those of group C (P<0.01) after modeling 2 days, but at the day 21.5 of pregnancy, mean arterial pressure, systolic pressure and diastolic pressure of group A were significantly decreased compared to group C (P<0.01). In groups A and B urine TP/creatinine ratio were significantly higher than those of groups C and D(P<0.01), also their liver, kidney and placenta tissue structure get pathological changes, and 70.49% fetal growth were ceased in groups A and B. Conclusion Preeclampsia model rats have been established successfully. This rat model will contribute to research of preeclampsia.

Mouse Model for Fetal Growth Restriction in Pregnant Metaphase by Limited Feeding

LIU Xue-xu, YANG Wen, DING Yun-ping

2015, 35(6): 453-457. DOI: 10.3969/j.issn.1674-5817.2015.06.005

Asbtract ( 292 )

PDF (450KB) ( 352 )

References | Related Articles | Metrics

Objectives To established the fetal growth restriction (FGR) mouse model in pregnant metaphase by limited feeding. Methods Ninety KM mice (60 females and 30 males, 3 months old) were divided randomly into 5 groups consisting of the control, model I, II, III and IV group. Mice in each group (12 female and 6 male) were placed in the mating cage with one male and 2 females. All the pregnant mice were fed ad libitum before day 10 of pregnancy. After that, model I, II, III and IV group were fed with 80%, 70%, 60% and 50% of daily ration respectively according to control group feeding ad libitum. Results The average weight increment in 15-day pregnancy mice was 26.03 g, 21.31 g, 19.09 g, 16.96 g and 13.19 g in control, model I, II, III and IV group, respectively. The weight increment in model group was significantly higher than that of control group (model II group P<0.05, III group and IV group P<0.01); The average litter weight was 21.12 g, 19.20 g, 18.95 g, 15.59 g and 12.99 g in control, model I, II, III and IV group, respectively. The litter weight in model group was significantly lower than that of control group (model III group P<0.05, IV group P<0.01); The incidence of FGR in model I, II, III, IV and control group was 42.97%、66.17%、66.38%、54.26% and 4.76%, respectively. The incidence in all model groups was significantly higher than that in control group (P<0.01). Except for the brain tissue in all model groups as well as the heart and liver tissue in model IV group, all the fetal organ weight in model group was significantly lower than that in control group (P<0.05). Conclusion It is feasible to establish the FGR model in pregnant metaphase by limited feeding. The best FGR model may be obtained by feeding the pregnant mice at 70% to 60% of daily ration of normal pregnant mice.

Effect of Implantable Vagus Nerve Stimulator on Epilepsy Models Induced by Kainic Acid in Rabbit

YIN Yin, ZHOU Hui-ying, XIAO Chun-lan, ZHANG Xin-guo, ZHOU Zheng-yu, WANG Jing

2015, 35(6): 462-466. DOI: 10.3969/j.issn.1674-5817.2015.06.007

Asbtract ( 238 )

PDF (1001KB) ( 325 )

References | Related Articles | Metrics

Objectives To study the intervention effect of implantable vagus nerve-stimulator (VNS) on epileptic rabbit model induced by kainic acid (KA). Methods The epileptic model of adult rabbit was established through the micro-injection of kainic acid into the cerebral cortex of animal. To relieve the epilepsy by VNS, the spiral electrode was placed on the vagus nerve, while the stimulator was implanted in subcutaneous tissue of the left animal back. Then 18 rabbits were randomly divided into 3 groups including epilepsy model group (group A, n=6), VNS pretreatment group (group B, n=6) and the VNS treatment group (group C, n=6). In group A, none VNS was implanted in the epileptic animal. In group B and C, VNSs were implanted 7 days before the KA injection for the epilepsy induction. The electricity stimulation was performed 2 hours before epilepsy in group B. However, stimulation carried out immediately when epilepsy emerges in group C. Epileptic behaviors and electroencephalography were observed and tested subsequently in all groups. The subcutaneous tissues around VNS were also observed via H.E stain to show the biocompatibility of VNS. Results All the animals showed stable epileptic seizure induced by KA and survived after the VNS plantation without obvious anomaly. The epileptiform electrocorticogram were ameliorated and the epileptic seizure remarkably suppressed in group B and were slightly restrained in group C. The tissues around the implanted VNS showed no obvious distinctions comparing with normal animal tissues in H.E stain. Conclusion VNS could alleviate the seizure of epileptic animal and show better effect when the vagus nerve is stimulated ahead of time. The implantable VNS is also biocompatible with animal subcutaneous tissues.

Establishment of Nested PCR Detection Method for Simian Cytomegalovirus and Preliminary Application

WANG Sha-sha, HE Zheng-ming

2015, 35(6): 467-472. DOI: 10.3969/j.issn.1674-5817.2015.06.008

Asbtract ( 334 )

PDF (1204KB) ( 348 )

References | Related Articles | Metrics

Objectives To establish simian cytomegalovirus (SCMV) specific nested PCR detection method, and to investigate SCMV infectious status in monkeys and related biological products. Methods Designed primers based on SCMV sequences and different monkey species and biological products were used to optimize the experiment. Results The established nested PCR detection method for SCMV can distinct SCMV from HCMV, MCMV, HSV and CHV, the test sensitivity is to viral DNA 18 pg/mL. By this nested PCR method, SCMV positive results were detected in monkeys and biological products, which suggested the high infection rate existed in monkeys and its biological products. Conclusions Nested PCR detection method has a good accuracy and sensitivity. It is found that high prevalence in Chinese primate colony. It is necessary to enhance detection for SCMV in monkeys and relevant biological products and to avoid potential risk of infecting in human.

Establishment and Preliminary Application of ELISA in Detecting Lymphocytic Choriomeningitis Virus Antibody in Mongolian Gerbil

WANG Ji, WEI Li, FU Rui, LI Xiao-bo, WANG Shu-jing, XING Jin, FENG Yu-fang, GONG Wei, YUE Bing-fei, HE Zheng-ming

2015, 35(6): 473-477. DOI: 10.3969/j.issn.1674-5817.2015.06.009

Asbtract ( 254 )

PDF (458KB) ( 378 )

References | Related Articles | Metrics

Objectives To develope the ELISA method for determination of lymphocytic choriomeningitis virus (LCMV) antibody in Mongolian gerbils. Methods Raised the Vero cell, vaccinated the LCMV, prepared the normal Vero antigen and specific LCMV antigen, titrated the best working density of enzyme union, the normal and specific antigen and verified the specificity, sensitivity, accuracy and stability of the method. Result The best working density of normal, specific antigen and the enzyme union were 0.4 mg/mL, 10 mg/mL and 1∶5 000, respectively. The inter-assay coefficient of variation of normal antigen and specific antigen were 6.8% and 8.9%, respectively, the intra-assay average coefficient of variation were 9.3% and 8.4%, respectively. The detection sensitivity was 1∶1 280. There was no cross-reactivity with mammalian orthoreovirus 3 (Reo3) and murine encephalomyelitis virus (M-TMEV). The stability test shows the relative deviation was below 25%. Conclusion The ELISA method is good in specificity, sensitivity, duplication, and stability, so ELISA could be used in detecting the LCMV antibody.

Construction and Evaluation of Chromosome Substitution Strains

ZHAO Ying, ZHAO Li-ya, CHAO Tian-zhu, ZHANG Rong, XING Zheng-hong, CHEN Guo-qiang, XIAO Jun-hua

2015, 35(6): 478-483. DOI: 10.3969/j.issn.1674-5817.2015.06.010

Asbtract ( 297 )

PDF (583KB) ( 372 )

References | Related Articles | Metrics

Objectives To shorten the fine-structure mapping time, construct and evaluate the programme of chromosome substitution strains (CSSs) . Methods The rst step required making hybrids between C57BL/6 male mice and donor female mice. N1 hybrids male mice were backcrossed to host C57BL/6 female mice. Progeny with a non-recombinant chromosome derived from donor strain, in this case chromosome 1, were identified in this and subsequent backcrosses. N2 Male mice were backcrossed to host C57BL/6 female mice at each generation. A certain number of progeny per generation was screened with short tandem repeat (STR) and single nucleotide polymorphism (SNP). These mice were backcrossed to host C57BL/6 at each generation until tenth generation. At the tenth backcross generation males and females with the non-recombinant chromosome derived from donor mice were intercrossed. Progeny of this intercross that were homozygous (homosomic) for chromosome 1 were used to propagate the homosomic strain. The homozygosity of CSSs was identified through scanning the whole genomic by microarray to compare with expected proportion. Results We have constructed two CSSs by the programme that had been constructed. The actual homozygosity of progeny with microarray was approaching the expected. Conclusion The constructed CSSs confirmed the replacement of target chromosome and over 99.59% chromosome segments from host C57BL/6. This breeding program to generate a CSSs panel proved to be feasible.

Study on Cryopreservation of Flow-cytometrically Sorted X-or Y- chromo some Bearing Sperm on Labrador Retriever

GAO Yi-long, QIN Hai-bin, WEN Hai, QIANG Jing-ning, HE Xing-liang, ZHANG Hui-dong, MA Da-jun, ZHU Qian, LI Gang

2015, 35(6): 484-489. DOI: 10.3969/j.issn.1674-5817.2015.06.011

Asbtract ( 440 )

PDF (609KB) ( 358 )

References | Related Articles | Metrics

Objectives To investigate the application of flow cytometric technology in separating X or Y sperm from Labrador retriever, and to define a protocol for producing sex-sorted frozen semen. Methods The semen were collected from 8 Labrador retrievers by manual stimulation and transported in one hour, and then the semen were separated into X- and Y- chromosome by SX_MOFLO flow cytometric technology, a modified high-speed cell sorter, and finally frozen thawed. Sorted X- or Y-sperm and unsorted sperm as control were reanalyzed and quality assessed. Results The purities of flow sorted sex chromosomes in spermatozoa of three dogs ranged from 89% to 91%. The average separation velocity showed significantly differences between sorted X- sperm (4 260/s) and Y- sperm (3 930/s). The sex ratio showed significantly differences between frozen- thawed sorted X- or Y- chromosome bearing sperm (89.11%, 90.67%) and the control. The forward progressive motility was not significantly different between sorted sperm and the control. The survive index on sorted sperm was significantly lower (0.251±0.006\0.054±0.007, P<0.05) than the control (0.048±0.006). But the post thaw acrosome integrity on sorted sperm was significantly higher (84.66±1.22\84.01±1.23, P<0.05) than the control (77.00±0.98). Conclusion It was possible to obtain sorted frozen- thawed Labrador retriever sperm for AI by using flow cytometry techniques.

Development of Evaluation and Assessment System for Laboratory Animal Employees Training

YU Chen-lin, SUN Xiao-xi, Cai Li-ping, TANG Qiu, LI Li

2015, 35(6): 498-503. DOI: 10.3969/j.issn.1674-5817.2015.06.014

Asbtract ( 241 )

PDF (1425KB) ( 349 )

References | Related Articles | Metrics

Objectives To development a evaluation assessment system for laboratory animal employees training. Methods According to the evaluating requirements for laboratory animal employees training, the system based on ASP.NET4.0 was designed by Microsoft Visual Basic.NET computer coding language, SQLServer database, Dreamweaver8.0 and Visual Studio2010. Results The system, including customer category management, customer information management, assessment indicators and weight parameters project management, score input management, statistical analysis of test results management and on-line management, was simple and friendly in interface, and flexible in operation. Conclusions The system may be suitable for evaluating laboratory animal employees training.

Table of Content