Laboratory Animal Science

Antitumor Effect and Immune Ｒegulation Activity of Ursolic Acid on Hepatoma H22 in Mice

LI Yan-Hong, LI Xiao-Bo, LU Xue-Ying, GAO Jian-Feng, XIAO Xiang-Wen

2013, 30(05): 1-5.

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Abstract: Objective The aim of this study was to investigate the antitumor effect and immune regulation activity of Ursolic acid ( UA) on H22 in mice． Method The models of tumor-bearing mice were established by i． p． injection of hepatic cancer H22 and the mice were randomly divided into 5 groups，each groups given by i． p． The tumor inhibitory rate，the thymus index and spleen index were calculated，lymphocyte proliferation of spleenstimulated by ConA and LPS were detected by MTT，T cell subsets CD4 + ，CD8 + were detected by FCM and the levels of The cytokine IL-2，TNF-α， IL-4 in serum were detected by ELISA．Ｒesult In vivo，the inhibitory rate of UA significantly higher than that of the model group． Compared with the model group，spleen index of high dose UA group were significantly lower． The T and B lymphocyte proliferation of spleen were significantly higher． The CD4 + T cell subset and CD4 + /CD8 + T rate were significantly increased，but CD8 + T cells has no significant change． The cytokine IL-2，TNF-α in serum expression were promoted with high dose UA，meanwhile the IL-4 expression was reduced． Conclusion The results show that UA can inhibit tumor growth both in vitro and in vivo． It could play a role of antitumor activity in vivo by regulating the body’s abnormal immune function in tumor-bearing mice．

Effect of Heroin on Bax Expression in Cerebellar Cortex in Ｒat

XIE Ming-Ren, LI Zuo-Ping, YU Fa-Rong, LIAN Xiu-Zhen

2013, 30(05): 6-8.

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Abstract: Objective To study the effects of heroin on the Bax expression of cerebellar cortex in rats． Method Addiction model was copied by intramuscular injection of heroin． Bax expression of cerebellar cortex was detected by immunohistochemistry methods．Ｒesult Ｒats had significant withdrawal symptoms after continuous injection of heroin for 7 days; the Bax-positive expression cells in cerebellar cortex of rats were increased significantly than that of the control group ( P ＜ 0. 01) ． Conclusion Heroin has the effect of inducing Bax gene expression and damaged brain cells．

The Effect of 15-KETE on the Protein and mＲNA Expression of Kv1. 5，Kv2. 1 in Ｒat Pulmonary Arterial Smooth Cells

GUO Shou-Li, SUN Yan, ZHANG Yi-Fei, ZHU Da-Ling

2013, 30(05): 9-14.

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Abstract: Objective To observe the effect of Kv1. 5、Kv2. 1 channels in rat pulmonary arterial smooth cells on the process of pulmonary vasoconstriction induced by 15-KETE． Method Western blot and PCＲ were used to identify the effect of 15-KETE on expression of Kv1. 5、Kv2. 1 channels in cultured rat pulmonary arterial smooth muscle cells ( PASMCs) ．Ｒesult ( 1) The expressions of Kv1. 5 mＲNA and protein in rat PASMCs were significantly down-regulated by 15-KETE． ( 2) Theexpressions of Kv2. 1 mＲNA and protein in rat PASMCs were significantly down-regulated by 15- KETE． Conclusion Kv1. 5、Kv2. 1 channels are involved in vasoconstriction induced by 15-KETE in rat PA．

Effect of Jianpi Huashi Granule on Visceral Sensitivity and CＲF/CＲF Ｒeceptors in Brain of D-IBS Model Ｒat

DU Hai-Yan-1, WANG Ying-Han, ZHANG Xiao-Feng, JIN Ji-Wei, GUO Hong-Jie

2013, 30(05): 15-19.

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Abstract: Objective To explore the effects of Jianpi Huashi Granule on visceral sensitivity and CＲF /CＲF-Ｒ1 expression in brain of D-IBS model rat． Method Seventy-two male Sprague-Dawley rats were equally and randomly divided into six groups: normal group，model group，dicetel group，low-，medium-，and high-dose Jianpi Huashi Granule groups; D-IBS was induced by restraint stress combined with administration of sennae decoction; The visceral sensitivity of rats was evaluated by colorectal distention，and recorded the water injection rate when the score of abdominal withdrawal reflex( AWＲ) was 3． The expression of CＲF and CＲF-Ｒ1mＲNA in the brain was detected by PCＲ．Ｒesult Compared with normal group，the water injection rate of the model group showed a significant decrease when the score of AWＲ was 3( P ＜ 0. 01) ，and the expression of CＲF and CＲF-Ｒ1mＲNA in the brain significantly increased ( P ＜ 0. 01) ; compared with model group， the water injection rate of the dicetel group，medium-，and high-dose Jianpi Huashi Granule groups showed a significant increase when the score of AWＲ was 3( P ＜ 0. 01) ，and the expression of CＲF and CＲF-Ｒ1mＲNA in the brain significantly decreased( P ＜ 0. 01) ． Conclusion medium-，and high-dose Jianpi Huashi Granule groups reduces visceral sensitivity in the rats with DIBS possibly by modulating the expression of CＲF and CＲF-Ｒ1mＲNA in the brain．

DEN-induced Hepatic Fibrosis-carcinoma in Ｒats

LI Yan, LI Xiao-Di, LIAO Ming

2013, 30(05): 20-23.

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Abstract: Objective To establish diethylinitrosamine( DEN) -induced hepatic fibrosis-carcinoma model in rat and observe pathological features during the process of hepatic carcinoma． Method Sprague-Dawley rats were randomly divided into DEN group and normal control group． The rats of DEN group were administrated 0. 01% DEN solution in water for 18 weeks，and the control group received vehicle． The animals were sacrificed at the end of 4th，8th， 10th，12th，15th week， respectively． Liver tissue sections were stained with hematoxylin-eosin ( HE) ．Ｒesult In DEN group，on 4th week，spotty necrosis of hepatocytes and inflammatory cell infiltration occurred in hepatic lobule． On 8th week， severe focal necrosis and hepatic steatosis of hepatocytes were observed accompanied with fiberous tissue proliferation in portal area． Hepatic lobules were encysted and separated by hyperplastic collagen bundles，and the normal structure of lobules was destroyed，and pseudo lobules formed on 9—12th week． After 16th week， the remaining surviving rats all developed hepatocellular carcinoma． The structure of liver tissue was normal in control group． Conclusion The intermission of DEN administer for hepatocellular carcinoma induction can elevate survival rate of rats in the end of carcinoma-inducing experiment． It is an ideal animal model for dynamically observing the pathologic process of hepatocellular carcinoma development．

The Study on TBI Model Praperation with eCCI in Ｒats and the Change of Serum S100B and NSE Content

GUO Xin-Rong, WANG Rui-Hui, WU Tao

2013, 30(05): 24-27.

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Abstract: Objective To study the TBI rats model produced by eCCI and provide a model establishing method of easy operation，the controllable degree of damage，good repeatability for the TBI experimental study． Method 65 SD rats，which were randomly divided into blank control group A，model group B，model group C，model group D and sham-operation group E，were produced as the animal model of brain injury in different degree by eCCI according to different selected parameters． The speed was 3 m/s and the depth was 3 mm in model group B． The speed was 4 m/s and the depth was 3 mm in model group C． The speed was 5 m/s and the depth was 3 mm in model group D． The quality of the model was evaluated through detecting the dynamic changes of S100B and NSE by ELIASA in animal serum．Ｒesult The content of S100B and NSE in serum was increased on the first day， second day，third day after animal damaged in model group by ELISA． Between the model group and sham operation group，there were significantly different ( P ＜ 0. 01) ． In model group B，C and D，the content of S100B and NSE was increased significantly with the increase of hit ( P ＜ 0. 01) ． Conclusion By using eCCI TBI rat model can be established with regulation of hit speed and depth． The damage of model was related to the hit speed． The method of TBI model preparation has the advantages of easy operation and control，better repeatability，and can be used according to the need to make various animal model for the study．

The Effect of 15-KETE on the Cytosolic ［Ca2 +］i in Ｒat Pulmonary Arterial Smooth Cells

GUO Shou-Li, ZHANG Yi-Fei, LIU Ye, ZHU Da-Ling

2013, 30(05): 28-32.

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Abstract: Objective To explore the effect and source of 15-KETE on the cytosolic ［Ca² ⁺］i in rat pulmonary arterial smooth cells( PASMCs) ． Method PASMCs obtained with enzyme ( collagenase I plus elastase) dispersing method were cultured in the first passage in vitro． Cells with concentration of 2 × 10 ^{－ 5} cells /mL were seeded onto a glass coverslip in 6 well plate，and cultured for 24 h in the incubator at 37℃． After cell adherence，the glass coverslip was taken out from 6 well plate，placed in special-made chamber，and washed with D-Hanks solution for 3 times． Cells were loaded with Fluo-3 /AM ( final concentration 10μmol /L) ，followed in the incubator at 37℃ for 30 min，then washed with D-Hanks solution for 3 times，extracellular dye was washed out，the Fluo-3 /AM loaded cells were available for ［Ca^{2 +}］i measurement within 2h． Using laser-scanning confocal microscope technique，the effect of 15-KETE on cytosolic ［Ca^{2 +}］i in rat PASMCs was measured．Ｒesult ( 1) 15-KETE ( 1 × 10 － 8 ～ 1 × 10 － 6 mol /L) increased cytosolic ［Ca² ⁺］i of rat PASMCs in a concentration-dependent manner． ( 2) The L-type Ca^{2 +} channel blocker verapamil ( 10 － 6 mol /L) and Ca^{2 +} -free Krebs solution significantly inhibited the increase of cytosolic ［Ca² ⁺］i induced by 15-KETE． Conclusion 15-KETE may increase cytosolic ［Ca^{2 +}］i of rat PASMCs， and the source of Ca² ⁺ was from intercellular solution．

Study on the Effect of Infusion of Alloreactive Lymphocytes Induced in vitro to Induce Liver Transplantation Tolerance in Ｒats

CAI Yong, YU Jiu-Fei

2013, 30(05): 33-36.

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Abstract: Objective To investigate whether infusion of alloreactive lymphocytes induced in vitro can prolong the survival of liver allograft in rats，so as to find out a new method to induce liver transplantation tolerance． Method Lymphocytes from Lewis rat were used as stimulating cells，lymphocytes from BN rat were stimulated by these stimulating cells and expanded in vitro; the proliferation of BN lymphocytes were determined by CFSE method． The proliferated BN lymphocytes were used as idiotype vaccine to vaccinate BN rats，then liver allograft from Lewis rats were transplanted to BN rats．Ｒesult Alloreactive lymphocytes proliferated vigorously in vitro; infusion of alloreactive lymphocytes induced in vitro can prolong the survival of liver allograft significantly in rats． Conclusion infusion of alloreactive lymphocytes induced in vitro may be a new method to induce donor specific transplantation tolerance．

Effects of Pretreatment with Fentanyl Combine Scopolamine on Myocardial Ischemia and Ｒeperfusion Injury in Ｒats

ZHOU Jie, SANG Li-Li, MAO Jian-Nan, LI Xing-Han, XUE Zhi, LIU Xiao-Fan, MENG Yong

2013, 30(05): 37-40.

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Abstract: To study the protective effects of pretreatment with fentanyl combine scopolamine on myocardial ischemia and reperfusion injury in rats． Twenty-four SD rats were randomly divided into three groups( n = 8 in each group) : control group ( group c，acute myocardial ischemia and reperfusion injury group) 、group F ( fentanyl group，the rats intravenously injected with fentanyl 0. 05mg /kg at 10min before myocardial ischemia) and group F + D ( fentanyl + scopolamine group，the rats intravenously injected with fentanyl 0. 05mg /kg and scopolamine 0. 3mg /kg at 10min before myocardial ischemia) ． Heart rate ( HＲ) ，left ventricular systolic pressure ( LVSP) ，left ventricular end diastolic pressure ( LVEDP) ，maximum positive and minimum negative left ventricular pressure derivatives ( ± dp / dtmax ) were continuously monitored and recorded at baseline，30min after ischemia，30min and 120min after reperfusion process． Contents of superoxide dismutase ( SOD) ，malonyldialdehyde ( MDA) and NO level in artery blood of rats were detected after reperfusion 120min． There were no significant difference in HＲ and LVEDP at any measurement point during the experiment among all groups ( P ＞ 0. 05) ． LVSP and ± dp /dtmax were significantly higher in group F than in group C at 30min、120min after reperfusion ( P ＜ 0. 05 ) ，and these were as well significantly higher in group F + D than in group F( P ＜ 0. 05) ． The serum activity of SOD was no significant difference in all groups ( P ＞ 0. 05) ． And the serum concentration of MDA was significantly lower in group F + D than in group F and group C ( P ＜ 0. 05) ． The serum concentration of NO was significantly higher in group F and group F + D than in group C ( P ＜ 0. 05) ． The arrhythmia score was significantly lower in group F + D than in else groups( P ＜ 0. 05) ． The protective effects of pretreatment with fentanyl combine scopolamine on myocardial ischemia and reperfusion injury in rats was better than fentanyl single．

Preventive Effect of Pollen Pini Extract on Experimental Hyperlipoidemia in Ｒats

LIU Yu-Ling, WANG Jian-Wei, TONG Ji-Ming, LI Sai-Ying

2013, 30(05): 41-43.

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Abstract: Objective To evaluate preventive effect of pollen pini extract on experimental hyperlipoidemia in rats and offer evidence for application of pollen pini extract． Method Eighty four Wistar rats were randomly divided into normal control group，model group，0. 3，0. 15 and 0. 075 g /kg of pellen pini extract groups，simvastatin control group and solvent control group． The rats in normal control group were fed with normal feed． The rats in model group were fed with high fat diet and simultaneously received intragastric administration of distilled water． The rats in solvent control group were fed with high fat diet and simultaneously received intragastric administration grape seed oil． The animals in other groups were fed with high fat diet and simultaneously received intragastric administration the corresponding drug． 12 h after the last administration，the content of TC，TG，LDL-C and HDLC in serum were determined．Ｒesult In all doses of pollen pini extract groups，content of TC，TG，and LDL-C in serum were lower than that of model group ( P ＜ 0. 05—0. 01 ) ． In 0. 3 g /kg and 0. 15 g /kg of pollen pini extract groups，the atherogenic index were lower than that of the model group ( P ＜ 0. 05) ． Conclusion Pollen pini extracts have certain preventive function on experimental hyperlipidemia in rats．

Exploration on Scientific Management of the Laboratory Animals Barrier Housing System in Bengbu Medical College

LAO Quan-Heng, LIU Lei, LIU Min

2013, 30(05): 41-54.

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Abstract: With the development of the laboratory animal’s subjects，many laboratory animals unit invested heavily in creating a barrier housing system to improve animal experimental environment． According to the operation situation of barrier housing system in our school laboratory animal center，by reference to the relevant national laboratory animal’s standards，an effective operational guidance standard of barrier housing environment laboratory was established． In the view of the particularity in management of the barrier housing system in higher medical colleges and universities，this article summarized the following aspects: the rules and regulations，personnel training，animals，and facilities，that should be paid more attention to in the management of the opening animal laboratory． on the basis of practical experience，the manage level of barrier housing system in the medical colleges could be improved．

Blood Biochemical Tests of Senior Inbred Wuzhishan Miniature Pigs

HUANG Lei, MOU Yu-Lian, YANG Shu-Lin, WANG Chen, CHENG Yu-Zhu, XU Pan, FENG Shu-Tang

2013, 30(05): 44-47.

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Abstract: Objective To analysis the blood biochemical parameters influenced by age and gender of inbreed Wuzhishan miniature pigs． Method 34 6-month-old pigs ( 7 ♂，27 ♀) ，as well as 21 pigs of 5—6 years old( 8 ♂，13♀) from F17，F18 offsprings of inbred WZSP were selected． After 12 hours of fasting，blood was collected via the superior vena cava and biochemical indexes were analyzed by full-Automatic biochemical analyzer．Ｒesult In all the blood biochemical indexes，3 of them showed gender differences ( P ＜ 0. 05) between young pigs，while 7 of them showed gender differences ( P ＜ 0. 05) among aged pigs． Meanwhile 16 of the blood biochemical indexes showed age differences ( P ＜ 0. 05) ． Conclusion The blood biochemical indexes of WZSP inbred lines display a significant age difference ( P ＜ 0. 05) ． This provides a basic reference data for choosing the aged miniature pigs as human aging animal models．

Discussion on the Methods about Feeding Management of Guinea Pigs

ZUO Hong-Mei-1, XU Li-Ba, ZHANG Shou-Ya, GUAN Ye-Zhi, WANG Long

2013, 30(05): 48-51.

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Abstract: Animal feeding can influence the growth and development，physiological function and metabolism of experimental animals，thus affecting the results of the authenticity，accuracy and scientific． The guinea pig is one of the experiments use more animal，but because of its activities with anatomical characteristics of sensitive reaction，with some special biological characteristics and physiological anatomy． Therefore，there are some special requirements on guinea pig fed and management，by the majority of researchers concern and attention．

Advances in Natural Killer Cells’Ｒeceptors and Functions

YANG Chao, HAN Ling-Xia, CHEN Hong-Yan

2013, 30(05): 55-58.

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Abstract: The Natural killer ( NK) cell is a lymphocyte that consisted by several subgroups，it has different kinds of receptors and displays various functions． Although the NK cell has been traditionally classified as cells of the innate immune system，it shares many similarities with adaptive lymphocyte，especially with cytotoxicity T lymphocytes． It sways great influence on immune memory，natural killing，anti-viral and anti-tumor． In this assay， we reported the latest findings on the classification of NK cells as well as its receptors and functions．

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