Table of Content

31 December 2013, Volume 30 Issue 06

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Analysis of the Health Status of Beijing Laboratory Ｒodents in Spring of 2013 by Pathological Evaluation

YAO Hao, WANG Ji-Hong, PENG Yun, XUE Zhi-Xin, SONG Zhi-Qi, LIU Chun-Fa, YUAN Zhen, 吕Yue , FU Yong-Yao, ZHOU Yang, ZHU Ting, YANG Li-Feng, ZHOU Xiang-Mei, YIN Xiao-Min, ZHAO De-Ming

2013, 30(06):  1-5. 

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Abstract:Objective In May，2013，our lab has taken samples from 14 units to investigate the health status of Beijing laboratory rodents． Method The heart，liver，spleen，lung，kidney，large intestine and small intestine of animals were collected，and were put into 10% paraformaldehyde solution for fixation，and then two different sections were prepared for optical microscopy observation，including all paraffin specimens stained with H． E． and the frozen sections stained with Oil Ｒed O and PAS． Ｒesult Liver cell swelling，slight lung lesion and protein cast of kidney were main lesions which might affect the health of laboratory rodents． Conclusion The results showed that the vast majority of laboratory rodents were pathologically diagnosed as healthy animals． The result might be related to breeding and feeding environment and transport stress，and the improvement of management level can ameliorate the health status of laboratory rodents．

Construction of the Baculovirus Ｒecombinant Bacmid of Synthetic Macaque B Virus gB and gC Genes

YAN Xiang, CAO Yu-Hua, LIU Hui-Fang, ZENG Lin, SUI Li-Hua, CUI Xiao-Xia, SUN Zhao-Zeng, HE Jian-Bin

2013, 30(06):  6-8. 

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Abstract:Objective To obtain baculovirus recombinant bacmids of synthetic macaque B virus gB and gC genes． Method The gene fragments of synthetic gB and gC were cloned in the transposition plasmid pFastBac-HTA． The positive plasmids were transformed to the competent cells of DH10bac． The recombinant bacmids were selected by PCＲ． Ｒesult The recombinant bacmids of gB and gC genes were constructed successfully． Conclusion Method of expression of gB and gC genes in the baculovirus expression system is established．

Study on Two Kinds of Skin Mechanical Barrier Disturbance Model in Mice

LIU Wan, XU Jia-Zhi, LIU Xing-Li, JIE Yu-Huan

2013, 30(06):  9-12. 

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Abstract:Objective To evaluate two kinds of skin mechanical barrier disturbance model of mice based on the concentration of epidermal ceramides． ，thickness and tissue pathology． Method After the stratum cornea of mice were damaged using adhesive tape and mixture of acetone and ether respectively for five days，concentration of epidermal ceramides，thickness and pathology were examined． Ｒesult Concentration of epidermal ceramides and thickness in both model groups were lower than those in control． Pathological changes were much obvious in the adhesive tape treated group，although they were observed in both models． Conclusion Both of the two methods can cause skin mechanical barrier disturbance in mice． Skin mechanical barrier disturbance caused by adhesive tape was more reliable and easily to be achieved．

Effect of Laboratory Mice Number on Detection of Ｒecombinant Human Erythropoietin Activity in vivo

ZHANG Wei-Ting, WANG Ying, ZHANG Hong-Xia

2013, 30(06):  13-16. 

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Abstract:Objective In vivo biological activity of recombinant human erythropoietin is one of the key indicators of quality control． The aim of this study is to investigate the effect of laboratory mouse number on detection of recombinant human erythropoietin activity in vivo，so as to reduce the retest and false alarm rates，and control the quality risk． Method According to the method described in the Chinese Pharmacopoeia (3)，EPO standard and sample solutions were administrated at low，medium，and high doses respectively to mice by subcutaneous injection． Blood was collected from the orbital venous plexus four days after injection，and be detected． Number of mice in each group was designed as 3，4，6，9，and 12． Ｒesult Ｒetest rate，relative standard deviation， maximum error range，and false alarm rate of the 3 mice group were the highest，no matter what dose of EPO were injected; there were no differences in retest rate，relative standard deviation，and maximum error range between the 4 and 6 mice groups; retest rate，relative standard deviation，maximum error range，and false alarm rate in the 9 and 12 mice groups were the lowest among the experiments． Conclusion The optional number of mice is suggested to be 9 per group in determination of in vivo biological activity of EPO．

The Effect of Diet-induced Obesity on Ｒeproductive Capacity of the Ｒat

LI Chang-Yu, Xu-Rong, ZHU Hai-Yan, LIU Wei

2013, 30(06):  17-20. 

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Abstract:Objective To probe into the effect of diet-induced obesity on reproductive capacity of SD rat． Method 60 female SD rats aged 6 weeks were fed with high-fat and high-calorie feed for 2 weeks，and then 20 obesityprone rats were selected according to their weight as the obesity group and other 20 rats were selected as the control group． The rats in the obesity group were fed with high-fat and high-calorie feed for 6 weeks to induce obesity and the rats in the control group were fed with ordinary rat feed． Before mating the body weights were measured and Lee’s index was calculated，and the levels of total cholesterol ( TC)，triglyceride ( TG)，high density lipoprotein cholesterol (HDL-C)，low density lipoprotein cholesterol ( LDL-C)，leptin and estradiol ( E2 ) in serum were determined． These female rats were mated with the male rats for one week，and the pregnancy rate and litter size were checked． Ｒesult The pregnancy rate in rats of the obesity group was significantly lower than that in the control group (P ＜ 0. 01) ． After the feeding for six weeks，the weight and Lee’s index of the obesity group rats were significantly higher than those of the control group (P ＜ 0. 01)，and the serum levels of TC，TG，LDL-C and leptin in the obesity group were significantly higher than those in the control group (P ＜ 0. 01) ． Conclusion The high-fat and high-calorie diet-induced rat declined in reproductive capacity，which may be related to the elevation in the serum level of leptin．

Effect of EＲK1 /2 Signaling Pathway and Vascular Endothelium Cells on Pulmonary Artery Constriction in Chronic Hypoxic Ｒats Induced by 15-KETE

GUO Shou-Li, ZHANG Yi-Fei, HAO Tian-Jun, SUN Yan

2013, 30(06):  21-25. 

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Abstract:Objective To observe the effects of EＲK1 /2 signaling pathway and vascular endothelium cells on 15- KETE-induced chronic hypoxic pulmonary artery constriction in rats using organ bath technique． Method Sixteen healthy Wistar rats (220 ± 20) g were randomly divided into two groups ( n = 8 each group) : normoxia and hypoxia groups housed in fresh air ( FiO2 = 21% ) and hypoxic air ( FiO2 = 10% ) in a hypoxic box，respectively． Pulmonary arteries( PA) were extracted after 9 days and cut into rings ( 0. 5 ～ 1. 0mm in diameter and 3mm in length) for organ bath experiments． The ring tensions before and after 15-KETE induction were compared． Influences of EＲK1 /2 upstream kinase inhibitor U0126 and endothelium integrity on 15-KETE induced HPV were investigated． Ｒesult With concentration increasing from 0 to 10 － 6 mol /L，15-KETE increased PA rings tension gradually in a dose-dependent fashion，and significantly constricted PA rings from the hypoxic rats． Ｒesponse of the hypoxic rings were significantly greater than that of normoxic ones (P ＜ 0. 05) ． The ring tensions induced by 15- KETE in endothelium-intact PAs were significantly greater than that in endothelium-denuded ones ( P ＜ 0. 05 ) ． U0126 significantly reduced vasoconstriction induced by 15-KETE in both endothelium-intact and -denuded rings． ( P ＜ 0. 05 in both) ． Conclusion 15-KETE significantly constricted PA rings in hypoxic rats in a dose-dependent fashion; EＲK1 /2 signaling pathway and vascular endothelium cells may be involved in vasoconstriction induced by 15-KETE，and EＲK l /2 in pulmonary artery smooth muscle cells may also be involved in vasoconstriction induced by 15-KETE in rats．

Effect of 15-KETE on EＲK1 /2 Activity in Ｒat Pulmonary Artery Smooth Muscle Cells

GUO Shou-Li, ZHANG Yi-Fei, HAO Tian-Jun, SUN Yan

2013, 30(06):  26-30. 

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Abstract:Objective To investigate the effect of 15-ketoeicosatetretraenoic acid(15-KETE) on EＲK1 /2 activity in rat pulmonary artery smooth muscle cells ( PASMCs) ． Method PASMCs of SD rats were collected by type I collagen plus elastase digestion，and cultured in control and defined media． The effect of 15-KETE on EＲK1 /2 activity in rat pulmonary artery smooth muscle cells was investigated by Western blot． Ｒesult 15-KETE had no effect on EＲK1 /2’s expression and activity at 10 － 6 mol /L from 0. 5 h to 24 h． But it could increase the expression of p-EＲK1 /2 after 0. 5，1，2，4 and 8 h incubation ( P ＜ 0. 05 ) ; 15-KETE，at concentration of 10 － 8— 10 － 6 mol /L，could significantly increase the expression of p- EＲK1 /2 ( P ＜ 0. 05) ; PD98059 and U0126，two specific inhibitors of EＲK1 /2，could significantly inhibit the effect of 15-KETE ( at concentration of 10 － 6 mol /L) on EＲK1 /2 activity ( P ＜ 0. 05 )，when PD98059 was added at 20 and 50 μmol /L and U0126 at 0. 1 and 1 μmol /L respectively． Conclusion 15-KETE has no effect on EＲK1 /2 expression，but can activate EＲK1 /2 in PASMCs and induce its phosphorylation in a time- and concentration-dependent manner． PD98059 and U0126 can inhibit the effect of 15 － HETE on EＲK1 /2 phosphorylation．

Comparison of Main Organ Coefficients between SPF and Conventional Hartley Guinea Pigs

TAN Qiao-Yan, ZHONG Hai-Chao, LIU Sheng-Lai, YAN Jia-Rong, KUANG Shao-Song

2013, 30(06):  31-33. 

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Abstract:Objective To determine the main organ coefficients in SPF and conventional Hartley guinea pigs． Method A total of 60 SPF and conventional guinea pigs (30 male and 30 female) at the age of 2 months were sacrificed in this study，and 13 organs，including heart，liver，spleen，lung，kidney and others，were collected and weighted． The relative weights of the 13 organs were calculated． Ｒesult Body weight and the weights of heart， spleen，lung，kidney，adrenal gland，and uterus showed extremely significant differences ( P ＜ 0. 01 ) between the SPF and conventional females; there are different significantly ( P ＜ 0. 01 ) between the male SPF and conventional Hartley guinea pigs in the weights of heart，liver，spleen，lung，kidney，adrenal，thymus ，testis， and epididymis． Coefficients of heart，spleen，lung，kidney，adrenal gland，and uterus were difference ( P ＜ 0. 01) significantly between the female SPF and conventional Hartley guinea pigs． And the organ coefficients of heart，liver，spleen，lung，kidney，adrenal，thymus，testis and epididymis were significantly different ( P ＜ 0. 01) between SPF and conventional males． Conclusion Different environmental and breeding conditions have some influence on main organ coefficients in SPF and conventional Hartley guinea pigs．

Establishment of Advanced Atherosclerosis Model in Ｒabbits

SUN Qing, ZHANG Lu, GUO Shu-Wen, ZHENG Min, ZHANG Kuo, LIU Jian, WU Jing-Ping

2013, 30(06):  34-40. 

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Abstract:Objective To develop a rapid and efficient method to establish the advanced atherosclerosis model． Method Fifteen New Zealand white rabbits were randomized into common carotid arteries (CCAs) balloon injury group ( n = 8 ) and sham-operated group ( n = 7 )，and fed with high-fat diets for fourteen weeks． Pathological characteristics of CCAs and other vascular segments，thoracic aortae，aortic roots and abdominal aortae，were observed． To assess the size of plaques，the percentage of intima proliferation area in internal elastic lamina ( IEL) area，average intima thickness in the radius of IEL and maximal intima thickness in the radius of IEL were calculated． Severity of lesions was evaluated by determining the histological classification recommended by American Heart Association． The serum lipid profiles of animals before and after experiment are measured． Ｒesult The size of plaques in thoracic aortae，aortic roots and abdominal aortae was decreased after fourteen-week highfat diet feeding． Although the biggest plaque and fastest lesion progression rate were observed in thoracic aortae ， the intermediate lesions were still the main manner characterized by intimal small extracellular lipid pools，rarely developing advanced lesions characterized by lipid core formation． The size of plaques in CCAs injured by balloon was bigger significantly than that of thoracic arotae ( P ＜ 0. 01 )，which was characterized by advanced atherosclerotic lesions． Conclusion Combined with high-fat diet feeding，advanced atherosclerosis model characterized by lipid core can be established rapidly and efficiently by balloon injury．

A New Way of Drug Administration to Adult Frog

WANG Jia-Mei, CHEN Xiu-Ping, ZHU Hao-Jun, DING Jia-Tong, PENG Yu-Fa

2013, 30(06):  41-44. 

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Abstract:To explore a new way to administrate drugs to the adult frog． Adult tiger frogs ( Ｒana rugulosa) at 90- day-old were fed with a mixture of pesticide (Bt suspension fluid) and forage． The frogs were randomly divided into blank control and five dosage groups (1 000，2 150，4640，10 000，21 500 mg / kg) ． Four repeats were conducted for each group and 5 frogs were administrated in each repeat． The median lethal dose ( LD50 ) and body weight gain were recorded to evaluate the feasibility of oral gavage in adult frogs in acute toxicity test． The results showed the oral gavage can make the frogs get nutrition support and gain weight． Moreover，there was a strong positive relationship between dose and response，and the LD50 value was 6 691 mg / kg． Therefore，the adult frogs can be administrated with various drugs by oral gavage，which will provide a new workable method for frog toxicity test．

The Feeding Management of Infant Monkey after Weaning

CHEN Li-Xiong, WANG Jun-Bin, YANG Feng-Mei, ZHAO Yuan, LU Shuai-Yao, LI Yan-Yan, YU Wen-Hai, HE Zhan-Long

2013, 30(06):  45-47. 

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Abstract:To improve the survival rate of infant monkeys after weaning，we took some improving measures，such as weaning early，keeping two monkey living in one cage，feeding less amount with 4 － 5 meals daily，paying more attention to the temperature and humidity control，careful sterilization of the cage and instrument，and preventing diseases． These procedures were able to solve the key and different problems during artificial rearing infant monkeys．

Diagnosis and Treatment of 28 Corneal Disorders in Canine and Feline

LI Shou-Jie

2013, 30(06):  48-50. 

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Abstract: Twenty-eight cases in dogs and cats，diagnosed corneal disorders but clinical intraocular pressure normal，were collected in present study． Patients were treated with atomization fumigation，traditional Chinese medicine therapy，and classification differentiation treatment based on disease types and syndrome，and treatment outcome was also assessed． The result showed the success treatment of 85% in canine and feline corneal disorders． It is suggested that the combination therapy is an effective mode for treating corneal disorders in canine and feline with abnormal intraocular pressure．  

Present Status and Strategy for Education and Training of Laboratory Animal Staffs in Hunan Province

ZHOU Ping, XU Lin-Ben

2013, 30(06):  51-53. 

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Abstract:Present status and problems of laboratory animal staff education and training in Hunan province were investigated and analyzed． Strategies，improving understanding，mechanism of laboratory animal staff education and training，and construction of innovation team，are provided．