Laboratory Animal and Comparative Medicine

Currents Sensitive to Amiloride in Mouse Spermatogenic Cells

WANG Chang-Song-1, GAO Xiao-Hua-1, LU Liang-1, GONG Wei-2, CHENG Jie-1, GAO Rong-1, XIAO Hang-1

2006, 26(3): 131-133.

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Objective To observe epithelial sodium channels （ENaC） currents in spermatogenic cells. Method Epithelial sodium channels were obtained in acutely dissociated mouse spermatogenic cells using whole-cell patch clamp. Results The currents were evoked by 200 ms test pulses between-100?+40 mV in 10 mV increments from a holding potential （HP） of -50 mV. Test pulses elicited rapidly activating and non-inactivating Na⁺ current. Na⁺ currents amplitude were significantly reduced by adding Amiloride （1 μmol/L） to the external solution, and displayed two kinds of inhibition effects （the ratio of 58.49%±9.82%，31.44%±5.69%）. Conclusion The characteristics of such currents and sensitive to Amiloride all approved that these currents recorded from mouse spermatogenic cells were produced through epithelial sodium channels.

Establishment of a Highly Metastatic Model of Colorectal Cancer in Mice through in Vivo Selection

ZHANG Jie-1, 2 , YAO Ming-2, YAN Ming-Xia-2, XU Bing-2, LIU Lei-2, KONG Han-Wei-2, LUO Yu-Zhu-1

2006, 26(3): 134-139.

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Objective To establish a highly metastatic model of colorectal cancer in mice by using organ selection of metastasis. Methods Histologically intact C26 colorectal cancer tissues were subcu-taneously implanted into the NOD-SCID mice which is immunodeficiency. In order to gain visible metastatic focus, the tumor bearing mice were undertaken tumor resection in first selection when tumor reached 1.5 cm in diameter. Screening was conducted according to the method of subcutaneous implantation→lung metastasis →subcutaneous inoculation→lung metastasis. Then the same steps were duplicated in BALB/c mice. Tumorgenicity, invasion, metastasis and morphological characteristics of the inplanted tumors were studied by light microscopy, electron microscopy and flow cytometry. Results Highly metastatic models of mice colorectal cancer were obtained after organ screening. After 5 generations in vivo, the carcinoma metastasis rate accounted to 100 percent in NOD-SCID mice, and the metastases of lungs and lymph nodes were noted in all of animals. While only a lower metastasis rate was found when the tumor was subcutaneously implanted in natural mice at the first time. But a 100 percent lung metastatic rate was observed again after being passaged in vivo for 4 generations in natural mice. Conclusions We had established a highly metastasis colorectal cancer model of natural mice which was high, steady, and intuitionistic metastasis and had same biological characteristics of mice colorectal carcinoma. The result provided a useful tool for the study of metastatic mechanism and treatment of human colorectal carcinoma.

Effect of Lead Exposure via Placenta and Milk on Early Nervous Reflex and Locomotives of Offspring in Rat

LI Shou-Ming-1, YANG Fan-2, LIU Zhi-Yong-2, LIU Chang-Ying-2, DAI Li-Guang-2, TU Ying-E-2, GONG You-3

2006, 26(3): 140-143.

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Objective To discuss whether the filial generation rats could be injured in neural behaviour development from low dose of lead acetate which were exposed to the parent rat during its pregnancy. Methods: At the first day of the pregnancy, the rats were divided into five groups, the first and second group are early pregnancy rats with l00mg·kg^-1-d^-1 and 1000 mg·kg^-1-d^-1 lead acetate by gavage at 10 ml·kg^-1,total exposure 42 days, the third and fourth group are late pregnancy rat with 100 mg·kg^-1. d^-1 and 1000 mg·kg^-1·d^-1 lead acetate, total exposure 28 days and the control group with distilled water.The offspring rats of each groups were tested for surface righting,cliff avoidance and slop turning at 5th day after birth，for body righting reflex and forelimb handgrip at 10th day. Result The offspring rats of four doses group have observably effects in early nervous reflex and locomotives, increasing with the exposure dosage and time. Conclusion Lead acetate affects offspring’s early nervous reflex and early locomotives through the placenta and milk.

Insecticidal Activity of Twelve Novel Pyridine Ring Derivatives of Podophyllotoxin

LIU Yan-Qing, ZHANG Shou-Gang, CHENG Jie, XIAO Hang, GAO Rong

2006, 26(3): 144-147.

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Objective The bioactivity of twelve novel pyridine ring derivatives of podophyllotoxin were tested against 3rd instar larvae of Culex pipiens pallens. Methods Using WHO bioassay method. Results From twelve novel pyridine ring derivatives of podophyllotoxin, three compounds 4.4、4.5 and 4.6 have more higher toxicities than others, and the LC₅₀ were 1.66, 3.96 and 1.54 mg/L respectively. Conclusion Based on the data obtained in this investigation, it is possible that the dissimilarity in the structure of the analogues leads to their different bioactivity.

Effects of TNFα on Activity of Tyrosine Protein Kinase of Membrane Insulin Receptor of Muscle and Liver in Mouse

WANG Zhan-Ke-1, XU Lin-Shui-2, YANG Li-Ping-1, CHU Fang-3, XIONG Xiao-Ping-1, GUO Bao-Zhu-1

2006, 26(3): 148-151.

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Objective To study the effects of TNFa on activity of TPK（tyrosine protein kinase） of the membrane insulin receptor of the muscle and liver in KM mouse. Methods Eighty KM mouse were divided randomly to the TH group（TNFa high-dose group）, TL group（TNFa low-dose group）, T1 group（TNFa once-dose group） and C group（control group）.The activity of TPK of the membrane insulin receptor in the muscle and liver without and with insulin-stimulated in all groups were observed. Results l）The activity of the TPK of the membrane insulin receptor in the muscle and liver without insulin-stimulated of TH and TL group were both lower significantly than that of C group, and TH group were lower than that of TL group.2） The changes of activity of the TPK of the membrane insulin receptor in the muscle and liver caused by insulin-stimulated of TH and TL group were both lower significantly than that of C group,and TH group were lower than that of TL group. 3）There were no significant difference of TPK of the membrane insulin receptor in the muscle and liver of the T1 group and C group. Conclusion TNFa can suppress the activity of TPK of the membrane insulin receptor in muscle and liver, and the effects of TNFa depends on the dose and time of TNFa treatment.

Construction and Application of Eukaryotic Expression Vector of Mouse LIF

ZHU Jing-1, JIN Dong-Qing-1, LI Zhi-Yan-1, DOU Ru-Hai-1, YANG Na-Na-2

2006, 26(3): 152-154.

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Objective Constructing the eukaryotic expression vector of mouse LIF and conducing to inprove the directional differentiation research of ES cell about other laboratory animals. Method Mouse secretedLIF with and without signal peptide were cloned from mouse livers by RT-PCR, and then subcloned into pBS-T vector and pMD18-T simple vector. Digesting fragments were recovered and inserted into pSecTag/Hygro with molecular cloning technique, and digested analysis by restrictive enzymes. Results By digestion identification with restrictive enzymes the LIF gene was cloned into eukaryotic expression vector pSecTag/Hygro successfully and the exp. ession vectors we constructed were called pSecTag-lif（sp⁺） and pSecTag-lif（sp ）. Conclusion It is valid to construct the eukaryotic expression vector of mouse LIF, and this not only establishs the base of molecule principle that maintains the undifferentiated ES cell, but also can provide a new method for the research of transgenic laboratory animals.

Development and Application of A Novel Antiviral Biologic Products for Poultry

SUN Jing-Fang-1, LIU Xing-Fu-2

2006, 26(3): 155-159.

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Objective Based on recombination IFNA6 （Chicken interferon alpha）, anti-viral biologic products were developed, aiming at prevention and treatment of poultry viral disease. Method Chicken interferon alpha（IFN） and thymosin alpha（THY） were recombined and expressed in E.coli and Picha pas tor is. Results There were different IFN biological activity from fusion protein of Chicken interferon and thymosin （IFN-THY） expressing in different vectors and different strains, 0.13 X 10⁴-8.12 X 10⁴IU/ml at 96 hours, and 1.22 X 10⁵-3.27 X 10⁶ IU/ml at 110 hours. Conclusion There were significant THY biological activity from different samples at 48-110 hours.

Induction of Estrus in Beagle Bitches by Using PMSG and hCG

XING Hua-1, YU Dao-Lun-2, ZHOU Tian-Sheng-1, XUE Zheng-Feng-1, LI Hou-Da-1

2006, 26(3): 160-164.

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Objective To found efficient procedure of inducing estrus in the bitch using exogenous gonadotropic hormones. Method Three groups of Beagle bitch, including anestrum, young of nonestrus and sterile aged bitches, were studied by combination of pregnant mare serum gonadotrophin （PMSG） + human chorionic gonadotropin （hCG） with different dose and administration separately. Result PMSG + hCG procedure was effective in the groups of anestrus and young bitches of nonestrus but ineffective for the sterile aged bitches （68.8% 25.0%）. Conclusion The success rate of inducing estrus with low dosage and multiple administration is markedly higher than that of the bitches with one administration.

Elementary study on Genetic Diversity of Biochemical Gene Markers of Z:ZCLA Mongolian Gerbil

LIU Yue-Huan, KE Xian-Fu, LOU Qi, ZHOU Sha-Sang, SA Xiao-Ying

2006, 26(3): 165-168.

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Objective To probe into the genetic diversity of the conventional closed colony of Z:ZCLA Mongolian gerbil. Methods The polymorphism of the conventional closed colony of Z:ZCLA Mongolian gerbil（sample content=100 individual） was studied by 19 biochemical gene markers and recorded their characters. Results Among the 19 biochemical loci, The locus Es-1, CarL-2, Hbb, Gpi-1，Cs-1, Ce-2,Id-1,Mod-1 had only one allele, while the locus Es-2, Es-3, Es-4, Es-6, Es-8, Es-9, Es-10, Gpd, Pgm, Trf and Akp-1 had two to four alleles, respectively. Average allele number was recorded as 3.0. Conclusion The conventional closed colony of Z: ZCLA Mongolian gerbil has the medium genetic diversity.

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