Laboratory Animal Science

Techniques and Experience in Developing Animal Model for Study on Mice Optic Nerve Regeneration

FAN Zhi-gang~1,2,CHEN Dong-feng~2,GE Jian~1

2006, 23(01): 1-4.

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Objective To summarize techniques and experience in developing mice optic nerve crushing model for studying optic nerve regeneration.Methods 4 Bcl-2/GFAP-Tk double transgenic mice were obtained by mating male Bcl-2 transgenic mice with female mice expressing herpes simplex virus-thymidine kinase transgenic mice controlled under GFAP promoter(GFAP-Tk) and 4 Bcl-2 transgenic mice were used as controls,and ganciclovir was administered through slow-releasing osmotic pump to selectively eliminate activated astrocytes in Bcl-2/GFAP-Tk transgenic mice.The right optic nerve crush was carded out according to the standard procedure in these mice aged 8～12 weeks(20～30g) on the second day after implantation of the osmotic pump.The tissue samples were taken on the tenth day after the optic nerve crush,and the axon regeneration was analyzed qualitatively and quantitatively by GAP43 immunofluorescence staining.Rodamine conjugate CTB or replication defective adeno associate virus carrying enhanced green fluorescence protein(AAV-EGFP) was injected in vitreous to label axon anterogradely and to determine whether the regenerated axons reach the brain targets.Results The remarkable optic nerve regeneration and the growth cone-like structure were found and the regenerated axon count was 71.99±24.04 in Bcl-2/GFAP-Tk double transgenic mice after GCV administration and optic nerve crushing,but the axon reached into the brain was found.No axonal regeneration was identified in Bcl-2 transgenic mice.Conclusion Complete optic nerve crushing in the mice is an ideal mode for studying neural regeneration,but stable techniques of developing the mode and specific labeling methods for axonal regeneration are imperative.

Study on Developing the Standard Animal Model of Blood Deficiency Syndrome

YANG Lan,ZHU Bi-de,PENG Cheng

2006, 23(01): 5-9.

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Objective To develop the standard animal models of blood deficiency syndrome.Methods Three kinds of blood deficiency syndrome models were developed by blood loss,hemolysis,and arrest of bone marrow,and the symptoms,signs,and laboratory indexes related to hematogenesis were observed.The therapeutic effects of Siwutang(四物汤) were observed as counterevidence in three kinds of model. Results The manifestations of the animals in the three kinds of model were in keeping with clinical manifestations of blood deficiency syndrome, with the changes of peripheral blood,marrow cells and their cell-division period,and Siwutang(四物汤) could exert satisfactory therapeutic effects on the changes.Conclusion Three kinds of standard animal model of blood deficiency syndrome were established in mice.

The Expression of NF-kB p65 mRNA in Lung Tissue of Acute Lung Injury Caused by 'Two Hits' in Rat

CHEN Min-li,XU Xiao-ping,YING Hua-zhong,Lü Jian-min,WANG De-jun

2006, 23(01): 10-12.

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Objective To observe the change of nuclear factor-kappa B(NF-kB) p65 mRNA expression in the lung tissue with acute lung injury(ALI) induced by oleic acid(OA) and lipopolysaccharide(LPS) in rats,and to investigate the relationship between nuclear factor kappa B p65 and ALI.Methods OA(0.2ml/kg) and LPS(2mg/kg) were injected intravenously to Wistar rats to develop ALI.The lung index(LI),WBC count,and percentage of ploymorphonuclear leucocyte(PMN) were observed,and the expression level of NF-kB p65 mRNA in the lung tissue with ALI was determined by situ hybridization technique.Results WBC in peripheral blood,and lung index of the rats with ALI caused by the 'two hits' were significantly increased,and PMN percentage changed markedly with the significant decrease in neutrophils and the significant increase in lymphocytes.The pathological sections of the lung showed an inflammatory change.The expression of NF-KB p65 mRNA in the lung tissue of the rats with ALI was markedly enhanced. Conclusion The enhanced expression of NF-kB p65 mRNA may make important contributions to development of ALI by 'Two hits'.

Study on Part of Aging Indexes in Aged Male Rats

YANG Fei,HU Ying,YANG Ping

2006, 23(01): 13-16.

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Objective To do quantitatively studying on some important aging indexes in aged male rats.Methods T-SOD activity,MDA level in serum,E-2 and T levels in plasma,binding capacity of M receptor,bone density of proximal,middle and distal segments of left femur,and break bending load of right femur were determined in the rats aged 6,24,and 30 months.Results T-SOD activity in serum,and E-2 and T levels in plasma of the rats aged 30 months were significantly lower than those of the rats aged 6 months(P<0.01),and MDA level in serum of the rats aged 30 months was significantly higher than that of the rats aged 6 months.The binding capacity of M receptor,and the femur bone density in respective segments and the femur break bending load of the rats aged 30 months were significantly lower than those of the rats aged 24 months(P<0.05).Conclusion The male rats aged 30 months was visibly aged and weakened,and was used as the natural aging model.The senility of the brain was closely linked to senility of bone in male rats.

Abnormality of Colonic Electricity and Nitric Oxide in Rat with Ulcerative Colitis

ZHANG Yong-bin~1,ZOU Yi-hai~1,LIAN Zhi-chang~2,CHEN Wei-qing~2,LIU Xiao-qiu~2

2006, 23(01): 17-19.

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Objective To establish a rat ulcerative colitis(UC) model and observe the abnormality of its colonic electricity and nitric oxide.Methods The rat-models of ulcerative colitis were induced by 2,4-dinitrochlorobenzene(DNCB),an electrode was buried in rats' colon and their colonic electricity was recorded.The amount of nitric oxide(NO) in colon can be detected by the concentration of nitrite in the colonic liquid.Results It was found that colon of rat-models developed typical ulceration and colonic electricity was abnormal,the basic electric rhythm(BER) of colon was slowed;extent of contractile electrical complex(CEC) was decreased and the percentage of BER and CEC loaded with peak electricity were markedly lowered,the nitrite concentration was higher than the healthy and negatively correlated with colonic electricity.Conclusion It is revealed that nitric oxide may play an important role in the occurrence of ulcerative colitis in rat and is closely related to the pathologic changes and motility of the colon.

Breeding of the SPF Rabbits and Observations of Their Growing Characteristics After Giveing Normal Intestinal Microflora

ZHANG Kuo~1,WANG Yin-huai~2,ZHOU Shu-pei~1,KANG Ai-jun~1,TIAN Feng~1,ZHENG Zhen-hui~1

2006, 23(01): 20-23.

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Objective To establish the SPF rabbits breeding stock for our department,and to meet the needs of scientific research,production and test.Methods The rabbits were cleaned by cesarotomy,the young rabbits were reared in isolators by artificial breast feed.The young rabbits weaned were given with normal intestinal bacteria,such as bifidobacterium and Fusobacterium,then transfered them into the barrier system.Growing characteristics are compared between rabbits given with normal intestines microflora and non-intestines microflora rabbits.Results They were accorded with the standard of SPF rabbits of China,with microorganism of the rabbits examined.The body weights of the former increases significantly more than the latter.Conclusions The SPF rabbit breeding stock was established by cesarotomy in our department.

The Change of Pulmonary Compliance After Cold Preservation in Dexamethasone Solution

LUO Li-rong,CUI Ze-zhong,ZENG Wei-cheng,LIN Yan,YI Gang,SU Fang,FENG Zhi-qiang

2006, 23(01): 24-26.

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Objective To investigate the change of pulmonary compliance after cold preservation in dexamethasone solution in an isolated lung model.Methods 32 rabbit lungs were randomly divided into two groups.The treated group was divided into 3 parts,preserved for 24h,72h and 120h in dexamethasone solution at 4℃.In the control group,pulmonary compliance was measured at room temperature.In the treated group,after rewarm at room temperature,the lungs were placed in saline solution.And then compliance with pulmonary volume 10mL,20mL and 30mL was measured respectively during infusing with air (saline solution) and drawing air(saline solution).Results Compared to the control group there were no significant differences in compliance of cold-preserved lungs for 24h with infusing with and drawing saline solution,cold-preserved lungs for 72h with infusing with and drawing air with pulmonary volume 10mL,20mL and cold-preserved lungs for 120h with infusing with and drawing air with pulmonary volume 10mL,20mL(P>0.05),but were in that of other treated groups(P<0.05).Conclusion Pulmonary compliance after cold preservation in dexamethasone solution increased,and the effect gradually increased with cold preservation prolongation.

Analysis of Application of Pentobarbital Sodium to Anesthesia of Laboratory Rabbits

PENG Chang-ling,ZHU Yu-hua,ZHU Xiao-rong

2006, 23(01): 51-52.

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Objective To compare the effect of pentobarbital sodium injected in several ways.Method Rabbits were injected with 3% pentobarbital sodium intravenously,intramuscularly,peritoneally and subcutaneously.And the dose of each method was 30 mg/kg,30 mg/kg,30 mg/kg and 40 mg/kg.Time of anesthetizing and sleeping were recorded and also eyelash reflex,reaction of stion,body temperature,breathing frequency,and the time of recover after being anesthetized were recorded.The experiment data were collected.Results The range of anesthetizing time,sleeping time and recover time were 0～17 minutes,46～89 minutes and 26～41 minutes,respectively.Conclusion Researchers may choose different method to anesthetize laboratory rabbits with the dose given in the article.

Comprehensive Control Measures on Parasite Diseases of Scaled-bred Macaques

HE Zhan-long~1,YANG Liang-yu~2,LU Shuai-yao,ZHAO Yuan,TANG Yong-ming,LI Shi-cheng

2006, 23(01): 53-54.

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It is necessary to develop scientific and comprehensive control measures on parasite diseases of scaled-bred macaques to improve quantity and quality of experimental macaques,which determines reliability of experimental results.

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