Table of Content

25 December 2011, Volume 31 Issue 6

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Down-regulated Expression of Perilipin in Leydig Cells of Aging Male Rats

HE Hua, LIU Yuan-yuan, LI Yu-hua, YUAN Wei, LI Runsheng

2011, 31(6):  405-409.  DOI: 10.3969/j.issn.1674-5817.2011.06.001

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Objective To evaluate the relationship of Perilipin expression with reduced androgen synthesis in aging male rats. Methods The three-month old rats were injected every day with D-galactose (300 mg/kg) for two months to establish the model of aging rat. The levels of total testosterone (TT), superoxitase dismutase (SOD) and malondialdehyde (MDA) in serum were used as the criterion to evaluate the quality of the animal model. For measurement of differentially-expressed genes, the Leydig cells was separated by Percoll discontinuous density gradient centrifugation, cultured for three day and then used for preparation of total RNA. Gene expression profile in the aging Leydig cells was studied in by cDNA microarray technique plus validation of screened genes by RT-PCR. Results In D-galactose-injected rats, the serum levels of MDA and SOD were significantly higher than those of the control rats. However, the serum level of TT was significantly lower in the rats injected with D-galactose than that of the control rats. Expression of hundreds of genes changed in the Leydig cells of aging rats, compared to that in the control, screened by cDNA microarray technique. Among these genes, Perilipin expression was selected for validation by RT-PCR technique, and our data indicated that levels of both Perilipin A and Perilipin B expression were significantly reduced in the aging Leydig cells. Conclusion The aging male rats were well established. Down-regulated expression of Perilipin may result in a reduced androgen synthesis in Leydig cells of aging male rats.



Establishment of Otitis Media with Effusion Induced by Lipopolysaccharides in Rat

LI Xiang-cui, ZHANG Yi, LI Ji-ping, WANG Jia-dong

2011, 31(6):  410-413.  DOI: 10.3969/j.issn.1674-5817.2011.06.002

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Objective To establishing the rat model of otitis media with effusion (OME). Methods The rats were inoculated with lipopolysaccharides (LPS) into the middle ear cavity through tympanic membrane. The pathologic changes of middle ear cavity and mucocilliary transporting system on the tympanic ostium eustachian tube were observed by light microscope and scan electrical microscope on days 1, 3, 7 after injecting LPS, respectively. Results Effusion were observed on day 1 after injecting LPS through tympanic membrane and disappeared after 7 days through otoscope.The pathologic changes were observed most severe on day 1 and returned to normal at 7 days under light microscope, and the impairment of mucociliary transport system such as lodging, shedding of cilias were observed on day 1 after injecting LPS and returned to normal after 7 days with scan electrical microscope. Conclusions Lipopolysaccharides (LPS) inoculated into the middle ear cavity through tympanic membrane can induce the inflammatory response and effusion of the middle ear, which can effectively establish the rat model of otitis media with effusion.



Establishment and Assessment of Gastric Mucosal Lesion Model in Rabbits Induced by Different Dossages of Anhydrous Ethanol

ZHANG Xuan, GAO Yan, LI Ning-shan, DING Xia-juan, XIONG Chun-yan, ZOU Jian-zhong, DU yong-hong

2011, 31(6):  414-417.  DOI: 10.3969/j.issn.1674-5817.2011.06.003

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Objective To observe the degree of injuries in rabbit gastric mucosal induced by different doses of anhydrous ethanol. Methods Twenty-eight New Zealand White rabbits were randomly divided into A, B, C, D groups (n=7). Group A was control group. The anhydrous ethanol 2.35 ml/kg , 1.00 ml/kg, 0.50 ml/kg with the dossage of were taken by intragastric administration to induce gastric mucosal lesion in group B, C, D respectively . The degree of gastric mucosal lesion was assessed by general morphology, biopsy and general physical condition. Results Compared with A group, gastric mucosal lesions were observed in the other three groups.Through general morphology of gastric, gastric ulcers were observed in group B, hyperemia, edema and erosion of gastric mucosal were observed in group C, but no significant lesion was observed in group D. Through biopsy, the lesions in group B were observed in muscle, mucosa in group C and nucosa epithelial in group D. Conclusion Anhydrous ethanol can induce gastric mucosal lesion models, 2.35ml/kg can induce gastric ulcer model, 1.00 ml/kg can induce gastric mucosal lesion model of erosion and hemorrhage, and 0.5ml/kg dosage did not show significant effects on gastric lesion.



The Effect of Epigalloca-techin-3-gallate(EGCG) on Expression of GLUT4 in Skeletal Muscle Tissue of Goto-kakizaki Rats

ZHAO Xian-zhe, ZHANG Jiang, QIAO Wei-wei, CHEN Li-xin, GU Jian-zhongzhong

2011, 31(6):  418-421.  DOI: 10.3969/j.issn.1674-5817.2011.06.004

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Objective To investigate the effect of epigalloca-techin-3-gallate(EGCG)on expression of GLUT4 in the skeletal muscle tissue of Goto-kakizaki rats. Methods Goto-kakizaki diabetic rats (GK, n=40) were observed and Wistar rat (n=10) as control. Rats were randomly divided into fIve groups: normal control group, TIIDM model group , diabetic model plus EGCG of 50mg/kg, 100mg/kg and 300mg/kg therapy group .After 6 weeks,blood glucose,serum lipids and the expression of GLUT4 were detected. Results EGCG obviously improved the OGTT and serum lipids, and increased the expression of GLUT4 in skeletal muscle membrane fraction. Conclusion EGCG can decrease the levels of blood glucose in type II diabetic rats, and its mechanisms might be closely related to the elevation of the level of GLUT4 in the skeletalmuscle tissues of rats.



Effects of Hypoxic Training on Blood EPO, Hb and RBC Level in Rats

LOU Shu-jie, JIAO Ying

2011, 31(6):  422-425.  DOI: 10.3969/j.issn.1674-5817.2011.06.005

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Objective To investigate effects of 7 day hypoxic training on the level of Erythropoietin(EPO), hemoglobin(Hb) and red blood cell (RBC) in blood of SD rats, and to obtain the experimental evidence of adaptation to hypoxic training in rats. Methods The rats were exposed in different concentration of hypoxic environment for 2h/day, the time of treadmill running of rats is 30min/day. The level of EPO was checked by ELISA, and the level of RBC and Hb were measured by automatic blood cell analyzer. Results (1) No effects of hypoxic exposure in 15% O₂ and 11% O₂ environment on EPO level were found significantly, but treadmill running in 15% O₂ environment enhance EPO level in blood of rats. (2) The number of RBC was increased significantly in rats which exercised on treadmill or stayed in 15% O₂ or 11% O₂ environment. (3) The increase of concentration of Hb was found significantly in rats which stayed in 15% O₂ or 11% O₂ environment. (4) Effects of hypoxic training on RBC and Hb level disappear in rats 15days after hypoxic training. Conclusion Production of RBC and Hb are significantly induced by hypoxic training in rats, but lasting time of enhancing effect is shorter than 15 days.

Inhibitory Effects of Sporoderm-broken Ganoderma Lucidum Spores on T-cell lymphoma

CHEN Juan, YU Ying-hao

2011, 31(6):  426-431.  DOI: 10.3969/j.issn.1674-5817.2011.06.006

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Objective To evaluate the inhibitory effects of sporoderm-broken Ganoderma Lucidum Spores on EL-4 cell in vitro, and in vivo, the nude mice model of the subcutaneous transplanting thymic lymphoma induced by MNU was established to evaluate the inhibitory effect. Mdthods The inhibitory effects of Ganoderma Lucidum Spores on EL-4 cell were detected by MTT method in vitro. The models of the subcutaneous transplanting tumor were established by Thymus T-cell lymphoma cells induced by MNU, the inhibitory rates were observed, and the tumor tissues of the nude mice were taken with HE staining. Results With the increasing of time, the IC₅₀ values were decreased, the minimums of which were 4.76g/L after 72 hours. In vivo, the Ganoderma Lucidum spores had certain experiment therapeutic anti-tumor action, and the inhibitory rate was 45.8% with the dose of 4g/kg. The morphologic study of Thymus lymphoma cells of Garoderma Lucidum Spoers group showed more necrosis and nucleoli crumb. Conclusion Garoderma Lucidum Spoers could play inhibitory action on lymphoma cell at high concentration and high dosage.



Effect of Anti-aging Tablets on Special Learning Memory and Free Radical Metabolism in Brain of Kidney Deficiency Mouse

JIA Lin-chao, LENG Xiao-xia, CHEN Min-li, PAN Yong-ming, SHOU Qi-yang, ZHOU Wei-min, TAO Tao, ZHU Ke-yan

2011, 31(6):  432-435.  DOI: 10.3969/j.issn.1674-5817.2011.06.007

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Objective To observe the effects of Anti-aging tablets on special learning memory and free radical metabolism in brain of kidney deficiency mouse. Methods Fifty-four male KM mice were categorized into six groups randomly: normal control group, kidney deficiency group (over-fatigue plus sexual overstrain group), low, middle, high doses of Anti-aging tablets groups and Golden Chamber Pill group. Over-fatigue model could be made by forcing mice to swim and kidney deficiency by colldege effect to induce sexual overstrain in male mice. Anti-aging tablets and Golden Chamber Pill could serve as medication. Changes of weight, water maze special learning and memory test, SOD activity and MDA content of brain were examined in different groups. Results The weight and SOD activity of brain in kidney deficiency group were lower than those in control groups (P<0.05, P<0.01), and the latency period of water maze and MDA content of brain were higher than those in control groups (P<0.01), while the weight and SOD activity of brain were higher in Anti-aging tablets groups and Golden Chamber Pill group (P<0.05,P<0.01), and the latency period of water maze and MDA content of brain were lower in Anti-aging tablets groups and Golden Chamber Pill group (P<0.05, P<0.01), compared with those in kidney deficiency group. Conclusion The Anti-aging tablets could be increased the activity of antioxidant enzymes and suppressed the generation of lipid peroxides, and which could be improved the memory ability of the mice.

Observation on Motions, Growth and Development after Tail Docking in Tree Shrew

LI Bo, JIN Hui-yan, CHEN Li-ling, ZHENG Hong, LIU Ru-wen, HE Bao-li, SHENG Pei-qing, JIAO Jian-lin

2011, 31(6):  436-440.  DOI: 10.3969/j.issn.1674-5817.2011.06.008

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Objective To explore the tail docking experiment in tree shrews and to observe the effects on their motions and reproduction conditions in the laboratory domestication. Methods Forty-five tree shrews were chosen and randomly divided into three groups: tree shrews in group 1 (bred for three months after tail docking, n=15) and group 2 ( bred for six months after tail docking, n=15) are tailed between the second and the third caudal vertebra or between the third and the forth caudal vertebra. Group 3 is the blank control group (n=15). The increases of gastrocnemius muscle tension of group 1 and 2 were compared with group 3. The structures of gastrocnemius of each group were detected by light microscope. Tree shrews in each group mated 1∶1( five males and females each group). Pregnancy, breeding and weaning were observed and recorded. Result All the tailed tree shrews have survived. The difference in the increases of gastronomies muscle tension in group 1 and 2 is significant compared with the blank control group (P<0.05). There are no obvious differences in the weights of the tree shrews and the increase of weights (P>0.05). No differences in conception rate were observed (P>0.05). Conclusion Tail docking has a certain inhibitory effect on the tree shrews’ motions, but they can grow, breeding and develop normally.



Determination of Cortisol in Plasma of Cynomolgus Monkeys by LC-MS/MS and Stress Monitoring

ZENG Xian-cheng, JI Long-feng, ZHANG Ling-li, YAN Dong-ming, ZHOU Chao, JIANG Zi-rui, PAN Xue-ying, CHANG Yan, MA Jing

2011, 31(6):  441-446.  DOI: 10.3969/j.issn.1674-5817.2011.06.009

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Objective To establish a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to determine cortisol levels in the plasma of cynomolgus monkeys and evaluate the influence of transportation, fasting and chair restraint on its levels in plasma. Methods Seven male monkeys were transported for eight hours by airplane and truck, and the blood samples were collected on day 1, 7, 14, 20 and 28 after transportation. The blood samples in six male monkeys were collected after the animals were fasted for 0, 8, 16 and 24 hours, and after one or five times of two-hour chair restraint. Cortisol was determined by using LC-MS/MS. Results The cortisol levels in plasma samples increased significantly after transportation, and then decreased gradually, and reached a stable level three weeks later. The cortisol levels in plasma samples were significantly increased after 8, 16 and 24 hours of fasting, but no differences were observed among the three types of fasting. The cortisol levels in plasma samples increased significantly after one and five times of chair restraint, but the increased extent of the latter was lower than the former. The calibration curves were linear over the concentration range of 20.0-2000.0 ng/ml. The intra- and inter-day precision was less than 4.94% and 5.69%, respectively, and the accuracy (RE) was less than 10.04% and 5.69%, respectively. The extraction recovery was between 70.3% and 74.7%. The analyte proved to be stable within three freeze-thaw tests and in the autosampler for ten hours at 20℃. Conclusion This method is rapid and specific and suited to cortisol determination in plasma. All of the stimuluses such as transportation, fasting and chair restraint were stressful for the monkeys.



Anatomical Observation on Pig Cardiac Blood Vessel Triangle

GUO Dong-jiao, CHEN Zhi-guo, QI Jiang-tao, QUE Dong-dong, CHEN Liu-ying, CHEN Xiao, YIN Zhong-wei, MA Jian-jun

2011, 31(6):  447-449.  DOI: 10.3969/j.issn.1674-5817.2011.06.010

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Objective To explore the anatomic structure of pig cardiac blood vessel triangle and provide anatomic data for animal models. Methods Fifty intact pig hearts were dissected to measure the location, quantity, outer diameter and angle of left coronary artery and its branches, as well as great cardiac vein and its tributaries. The characteristics of cardiac blood vessel triangle were analysed. Results The cardiac blood vessel triangle could be divided into intersectant type, associated type and complex type. In the triangle, the included angle consisting of anterior interventricular branch and circumflex branch was 97.0±15.4 , whose corresponding side length was 46.24±15.30 mm; the included angle consisting of anterior interventricular branch and great cardiac vein was 29.7±10.3, whose corresponding side length was 23.50±8.02 mm ; the included angle consisting of circumflex branch and great cardiac vein was 54.9±16.4, whose corresponding side length was 36.30±13.94 mm. There could be anterior branches of left ventricle and opposite angle branches in the triangle. Conclusions Pig cardiac blood vessel triangle is similar to that of human in form, but distinctly different in size. The pig can be used as excellent animal models for interventional treatments of coronary heart disease and coronary sinus.



Comparison on Organ Weight and Coefficient Between Conventional and Clean Cricetulus griseus

HOU Yan-yan, SHI Shen, MAI LI-kai, XU Xiao-hui, Muhebul, Dylinur, YUAN Jiang-ling, LIU Xiao-feng, GUI You-jun, LIAO Li-fu, YAN Shun-sheng

2011, 31(6):  450-452.  DOI: 10.3969/j.issn.1674-5817.2011.06.011

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Objective To detect and compare the differences in main organ weight and coefficient between conventional and clean Cricetulus griseus. Methods Forty conventional and clean Cricetulus griseus were dissected respectively. Weight of tissues and organs was measured with electronic balance to detect indices of 12 organs, and the organ coefficient were calculated. Result Compared with the conventional and clean male Cricetulus griseus, there was statistical significance in weight of heart, liver and epididymis (P<0.01), and in visceral coefficient of brain and kidney (P<0.01). Compared with the conventional and clean female Cricetulus griseus ,showed statistical significances in weight of heart and renal coefficient (P<0.05). In conventional Cricetulus griseus, spleen coefficient was different between male and female (P<0.05). In clean Cricetulus griseus, there was statistical significance in heart coefficient between male and female (P<0.01). Conclusions There are obvious differences in weight and coefficient of organs between clean and conventional Cricetulus griseus.

Application and Energy Conservation Methods Assessing in Laboratory Animal Facility by Energy Monitoring System

DING Xian-ming, ZHOU Rong-gui, DING Yang, JI Rong-qiang, QU Li-juan

2011, 31(6):  453-458.  DOI: 10.3969/j.issn.1674-5817.2011.06.012

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Objective To establish a method for energy montoring and assessing energy conservation methods in laboratory animal facilitiy. Methods Annual energy consumption situation of laboratory animal facilitiy was assessed by an energy monitoring system. The total energy consumption of electricity, steam, heat and cooling capacity, etc. were recorded and analysised . The square meter energy consumption of the facility was calculated and compared with other buildings in our campus. Two different steam supply ways, pipeline and electric steam boiler, were measured by the energy monitoring system. Results Consumption of electricity, steam, heat and cooling capacity were 1.725 GWh、12244.90 and 1.497 GWh, respectively. The total electric and steam consumption of the facility were 2.783 GWh and 12553.29 GJ. Compared with 4 different buildings monitored, the energy consumption of animal facility was 8.52 to 21.88 times more. The system showed steam consumption of two different supplying ways were 9925.00 GJ and 2651.00 GJ, respectively. Conclusion The energy monitoring system could real-time reflect the energy consumption of our laboratory animal facilities. It was availible for energy consumption statistics and energy-saving administration. According to our results, electric steam boiler was a more economical way compared with pipe supplying.



Establishment of Chronic Renal Failure Model in Beagle Dogs by Integrative Operation

LIN Ruo-qin, FU Xiao-qing, YANG Zhao-yang, ZHU Long, LI Hui-yong

2011, 31(6):  459-461.  DOI: 10.3969/j.issn.1674-5817.2011.06.013

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Objective To study the surgical method for establishing chronic renal failure model at azotaemic stage in beagle dogs. Methods The integrative operation was preformed by electric cautery of renal cortex and renal artery ligation in left kidney. Right kidney was removed one week later. Blood urea nitrogen and creatinine were detected 3 days after surgery and every week after surgery. Results The urea nitrogen was 7.21±0.72 mmol/L and the creatinine was 82.40±9.42 µmol/L before surgery, 30 days after surgery was 11.22±5.69 mmol/L and 125.55±68.50 µmol/L respectively. Both of their P- value were less than 0.05. Conclusions Ideal chronic renal failure model at azotaemic stage in beagle dogs can be obtained by the combination of electric cautery of left renal cortex and left renal artery ligation.

Research Progress on Establishment of Animal Model with Staphylococcus aureus Infection

YANG Yu-qin, DING Yi-yuan, PENG Xiu-hua, XUN Chun-hua, ZHOU Wen-jiang

2011, 31(6):  473-477.  DOI: 10.3969/j.issn.1674-5817.2011.06.018

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The purpose of the review was to summary the infection methods, mechanism, the suitable animals, sensitive indicators in establishment of Staphylococcus aureus (SA), and the present situation of the application in pharmacodynamics evaluation .It may be provides the literature and ideas for building SA infection animal model which is simpler, more economical and closer to clinical human SA infection.

Establishment of Induced Animal Model with Thymic Lymphoma

WANG Hong-yan, SONG Xiang-fu, CHEN Yu-bing, GONG Shou-liang

2011, 31(6):  478-482.  DOI: 10.3969/j.issn.1674-5817.2011.06.019

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Thymoma is the most common mediastinal tumor, its etiology and pathogenesis is unclear. It is an invasive important means to establish suitable animal model for studing its etiology, pathogenesis and treatment. In this paper, we review some thymoma tumor model induced by carcinogens such as physical, chemical and biological factors . At the same time their trigger mechanism and influenced factors were also reviewed Then we simply introduce the advantages of the induced thymoma model.