Laboratory Animal Science

Xenotransplantation Model of A Human Small Cell Lung Cancer Cell Line——NCI-H446 in Nude Mice

LIN Yun_xiu, LU Lian_huang, LIN Jin_juan,CHEN Ying_yu

2004, 21(03): 1-3.

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Objective To establish a successful xenotransplantation model of a human small cell lung cancer cell line (NCI_H446) in nude mice. Methods NCI_H446 cells were transplanted subcutaneously into nude mice.After the first generation,the xenografls were transplanted subcutaneously into nude mice with serial passages for three generations.The growth features,histology,ultramicroscope structure and karyology of the successfully transplanted tumors were observed. Results After four generations of transplantation the latent period was shorten, and the spontaneous regression was not observed.The growth curve of the tumor is like “S”.The karyotype of tumor cells belongs to human karyofype.

The Effect of Hairless Mutant Gene on The Thymus and Skin Under The C_(57)BL/6 Genetic Background

YAN Hong-xia, JIN Shu-xing, ZHANG Jin-tao, DU Chun-yan

2004, 21(03): 4-6.

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Objective To study how the mutation gene affects the thymus and skin of the C 57BL/6 hairless mice. Methods The procedure of shedding hair and histological structure on the thymus and skin of the C 57BL/6 hairless mice in 19 days were studied,and compared with C 57BL/6 mice of the same age. Results The C 57BL/6 hairless mice developed a normal coat up to the age of 12 days,but then lost all hair.At the age of 4 weeks the hairless-like mutant mice become essentially naked except for some vibrissae,and grew no new hair.With increased age,the mutant skin form folds and flaps and had rhinoceros-like appearance.The claws were noticeably elongated.Mutant mice had heavily pigmented skin,and had some small papules on the surface of skin.Histologically,There was keratotic cuticular layer in the epidermis.The hair follicle of C 57BL/6 hairless mice in 19 days lost shaft to distend to be lenient cavity, some follicles contained keratin fragments.The Remains of hair papilla became trapped deep in the dermis.Sebaceous glands were well developed.As for the morphology and structure of thymus in 19 days,there was no distinct difference between C 57BL/6 hairless mice and normal mice,but the thymus of hairless mice atrophy early than that of contrast mice. Conclusion The hairless mutant gene effects hair follicles cycle and thymus degeneration of the C 57BL/6 hairless mice.

Determination of Hematological and Biochemical Parameters of F_2 Mutant Hairless Mice

WANG Dong-ping, LI Shan-ru, LUAN Rong-hui, LI Gui-jun

2004, 21(03): 7-9.

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Mutant hairless mice with a first sparse coat appeared spontaneously in the production stock of normal BALB/c mice. For study of mutant hairless gene affected biological characteristics of the mice,hematological and blood biochemical index of F 2 mutant hairless mice were determined. The results showed there were a few significant differences in mice between sexes (male and female mice).There were significant differences Bun,Ca of mutant hairless mice.There were significant differences WBC,RBC,HCT,GPT.Alb,Mg of sparse-haired mice. The results showed there were significant differences between metabolize and electrolyte of F 2 mutant hairless mice and mutant hairless gene affected the metabolize infection.

Preliminary Studies for Biological Properties of IRM-1 Inbred Strain Mouse

WANG Yue-ying, WU Hong-ying, YUE Jing-yin, MU Chuan-jie

2004, 21(03): 10-12.

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Objective To obtain the data on characteristics of the generation and growth of IRM-1 inbred strain mouse. Methods The body weight and different organ weight from 6 and 9 week old IRM-1 mouse were weighed, then the visceral coefficient was counted. Results and conclusion Heart, liver and kidney weight of male mouse at 9 week old are higher than those of female. Heart, liver, kidney and lung weight of male mouse at 6 week old are higher than those of female, thymus weight of male mouse at 6 and 9 week old is lower then that of female. Properties of generation and growth of IRM-1 are similar to IMR-2 inbred strain mouse.

Study on Acute Immune Liver Injury Model of Rats Induced by BCG and LPS

QIU Ying-feng,MIAO Xiao-hui,WANG Jun-xue,ZHANG Rui-qi,CAI Xiong,NI Wu

2004, 21(03): 15-19.

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Objective To establish acute immune liver injury model of rats for studying pathogenesis of fulminant viral hepatitis. Method Acute immune liver injury model of rats were induced by injection of different dosages of BCG and LPS. Hepatic injury were assessed by the level of serum transaminase and pathological changes of liver. The percentage of CD3 +,CD4 + and CD8 + cell was determined by flow cytometer. Level of cytokines including TNF α, IL-6 and IL-10 in sera was detected by Enzyme-linked immunoadsordent assay. Result Acute immune liver injury model of rats was successfully established by ip injection of BCG 5×10 7 bacilli/rat and LPS 30 μg/kg body wt, 10 days and 8 h before the assay,respectively. The level of GPT and GOT increased significantly, and pathological changes of “Ⅲ” grade and “Ⅳ” grade were observed in more than 70% assessed liver tissue sections. Level of cytokines including TNF α,IL-6 and IL-10 in sera was higher than that of normal control group. Conclusion Acute immune liver injury model of rats can be established by combined injection of BCG and LPS. The model might be further applied in the research of fulminant viral hepatitis.

Study on IB-ELISA Diagnostic Reagent

LIU Jun 1 CHEN Fu-yong 2

2004, 21(03): 20-23.

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Purpose To establish a convenient,specific and rapid detective technique for routine measuring antibodies to infectious bronchitis virus.Method IBV was propagated in the allantoic cavity of 9-day-old specific pathogen free(SPF),as well as in the material of chicken embryo fibroblasts(CEF)by serial passage.Followed purification,the protein quantity was measured,which was used as coated antigen.Conjugation was labeled rabbit anti-chichen IgG by using periodate method,TMB and OPD as substrate,to set up enzyme-linked immunosorbent assay(ELISA)for testing infectious bronchitis virus antibodies.Results The above two obtained IBV could be used as coating antigen.No crossing reaction was observed with antibodies to Newcastle disease virus(NDV),Marek's disease virus(MDV),infectious bursal disease virus(IBDV),infectious laryngotracheitis virus(ILTV),Avian influenza virus(AIV),Avian parainfluenza virus(APIV),Reticuloendotheliosis virus(REV),Infectious Coryza(IC),Mycoplasma gallisepticum(MG),Mycoplasma synoviae(MS).The coincidence of result is good by comparing with the imported reagent of IB-ELISA produced by IDEXX.Conclusion:The IB-ELISA diagnostic reagent can be used to detect IBV-antibody.

Cryopreservation of Chicken Primordial Germ Cells at Stage 19 Under Different Preservative Density

XIAO Xiao-jun, QIN Jie, CAI Lin-lin, LI Bi-chun

2004, 21(03): 24-26.

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Isolating the primordial germ cells(PGCs)from gonads at stage 19 by Ficoll density gradient centrifugation.Cryopreservating the PGCs at difference freezing media density.The vitality of the frozen-thawed PGCs was determined by the Trypan blue exclusion method,than the PGCs were cultured in vitro.The result showed:at the same density,there is significant difference(P<0.05) or very significant difference(P<0.01) among the freezing media.At the same freezing medie,there is significant difference(P<0.05)or very significant difference(P<0.01) among the different density,the density more higher,the vitality more lower.When the thawed PGCs culture in vitro,it can propagation,form PGCs colony,subculture.

The Safety Evaluation of The Extracts from Zostera marina

WANG Yue-ling,SHAO Yong-li,WANG Wei, ZHU Jian

2004, 21(03): 27-29.

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Objective Toxicity experiments were carried out to evaluate the safety of the extracts from Zostera marina.Method Acute toxicity and accumulative toxicity tests,Ames test and bone marrow micronucleus test were in KM mice.Results There were no acute toxicity and no accumulative toxicity founded in the test in mice.The Ames test was negative and no mutagenesis was detected in the experiments mentioned above.Conclusion There is no toxicity and no mutagenicity in the extracts from Zostera marina.It suggests that we can use the extracts safely.

The Effect of Ginkgo Foliage Preparation in Great Doses on Antioxidation Indexes of Dog Plasma

LU jing, FENG Gao-qian, ZHANG Lei, ZHU Zi-xin, HUANG Pei-li

2004, 21(03): 30-31.

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Objective In order to further understand the pharmacological action of the ginkgo foliage preparation, the dogs made to take orally the great doses of ginkgo foliage preparation and antioxidation indexes of the dog plasma were measured. Methods Six dogs were selected and took orally once ginkgo foliage capsule at a dose of 100 mg/kg body weight. The 5.0mL blood was collected through the hypodermic vein of the forelimbs on selective time. By using conventional kit, antioxidation indexes were measured. Results The activation of SOD in the dog plasma reduced after giving the medicine, and especially significantly reduced between 7 hours and 24 hours. The content of MDA went down in 5 hours after giving the medicine, and then increased between 7 hours and 24 hours. Conolusion It is suggested, if the doses of ginkgo foliage preparation are beyond the normal limit, the capability of antioxidation abates, because ·O 2- produces in the body cumulates.

The Effect Factors of Transplantation After Human Tumor Cell Suspension Inoculated Subcutaneousely into Nude Mice

TIAN Qing-e, LI Ma-lin

2004, 21(03): 47-50.

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Objective To improve the successful establishment of human tumor nude mice transplantation model subcutaneousely,authors reviewed the method and the effect factors of human tumor cell suspension inoculated subcutaneousely into nude mice.Methods According to reports and authors′ experience,we review the method and the effect factors of human tumor cell suspension inoculated subcutaneousely into nude mice from tumor cell,nude mice,experimenter etc.Results We see the specific method of operation and the effect factors of transplantation.Conclusions Paying more attention to the method and the effect factors of human tumor cell suspension may benefit to establish human tumor nude mice transplantation model subcutaneousely.

An Experiment on The Examination of Tyzzer Organism

LI Sheng-li, LU Jing, WANG Ju

2004, 21(03): 58-59.

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To examine whether a rat group has been infected by Tyzzer organism,10 young rats and 10 adult rats are seleted to be examined through pathology and IFA mammers. 10 Mongolian gerbils are also used to do a supplementary experiment.The results of three mammers are accordant,there is no infection of Tyzzer organism in this group.

Affirmation of The Pregnant Day by Cohabiting Rats in The Ovulatory Period

ZENG Hai-yan,MA Guang-yu

2004, 21(03): 61-62.

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In order to affirm the pregnant time to so as the drug to be delivered,we should know the pregnant day 1 of rats,we cohabited the rats in the ovulatory time by examination of the vaginal smear and the second day was regarded as the pregnant day 1.There is no difference from the traditional way,however this method is easy and simple to grasp.

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