Table of Content

31 March 2009, Volume 29 Issue 3

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Establishment of Premenstrual Syndrome Liver-qi Invasion, Depression Rats Model and Macro-evaluation

WEI Sheng，ZHANG Hui-yun

2009, 29(3):  142-146. 

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Objective Improve the method of making model and macro-evaluation about Premenstrual Syndrome Liver-qi Invasion, Depression Rats Model, providing a “zheng” animal model for the research of mechanism on emotional disease. Methods Adopted ethology and improved vaginal smear to determine rats oestrous cycle，apply open-field method to filt similar scoring rats entering experiment, adjust stimulus intensity and persistence time, introduce open-field method to estimate rats macroscopic appearance. Results Liver-qi invasion rats，scores were much higher than the control while liver-qi depression rats，scores decreasing notably, the trend was retrieved after administration. Conclusion The rat model were consistent with clinical manifestation and pathological changes satisfactorily. Modeling method was reliable and mature with a better credibility and repeatability. Adopting open-field method could distinguish syndrome belongings of rats model clearly. Thus provided a direct evidence for differentiation of symptoms and signs on animal models.

The Comparative Studies on Spermatozoa Cryopreservation in C57BL/6J Mice by Various Cryoprotectants

JIA Qing1，LIU Li-jun2，GAO Juan1，YANG Bo3，YU Li-li2，XU Ping2，RUI Rong4

2009, 29(3):  147-152. 

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Objective To establish a more suitable method to frozen sperm of C57BL/6J by comparing in vitro fertilization rate of post- thawed C57BL/6J mice spermatozoa in various cryoprotectants. Methods To R18S3-based fluid, adding with the different concentrations of egg yolk and glycerine as cryoprotectants to frozen sperm from C57BL/6J and Y3F mice and using the same method of sperm recovery with the rate of in vitro fertilization to evaluate the effect of frozen sperm. Results R18S3 with liquid-based, add with the concentration of 5%，10%，15%，20%, 25% of yolk and centrifuged, then the supernatant as cryoprotectants, the rate of in vitro fertilization and frozen-thawed sperm of C57BL/ 6J mice is 18.7%, 15.6%，36.0%, 40.0%，29.7% respectively with a significant different. R18S3 with liquid-based,add with the concentration of 1%, 2%, 3%, 4%, 5%, 6% glycerol as cryoprotectants, the rates of in vitro fertilization of freeze-thawed sperm of C57BL/6J mice were 7.8%, 10.0%, 16.9%, 6.0%, 10.3%, 18.1% respectively with a significant difference.The supernatant of R18S3 +5%yolk after centrifugation +6% glycerol; the supernatant of R18S3 +15% yolk after centrifugation +2% glycerol; the supernatant of R18S3 +20% yolk after centrifugation +1% glycerol as cryoprotectants, the rates of in vitro fertilization of freeze-thawed sperm of C57BL/6J mice were 16.7%, 36.0% and 22.9% respectively with a significant difference. R18S3 as cryoprotectants, the rate of in vitro fertilization of freeze-thawed sperm of C57BL/6J mice is 25%, the rate of abnormal cleavage is 2.6%; the rate of in vitro fertilization of freeze-thawed sperm of Y3F mice is 52.2%, the rate of abnormal cleavage is 5.1%. Conclusion The supernatant of R18S3 +15%/20% yolk after centrifugation as cryoprotectants can raise the rate of in vitro fertilization of freeze-thawed sperm of C57BL/6J mice but adding with the different concentrations of glycerol can reduce the rate. The frozen effect of adding with both yolk and glycerol less than the effect of adding only yolk, but better than the effect of adding only glycerol.

A Modified Passive Smoking and Endo-tracheal Instillation of LPS Method for Establishment of COPD Models in Rats

HU Rong1，YAN Guo-feng2，JIANG Hong1，ZHU Ye-sen1

2009, 29(3):  153-156. 

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Objective To evaluate the efficacy of a modified method of passive smoking and endotracheal instillation of LPS for establishment of chronic obstructive pulmonary disease (COPD) model in rats. Methods 30 Sprague-Dawley(SD) rats were randomly divided into three groups; normal, model and modification groups, 10 rats each. The rats in normal group received normal feeding. The rats of model group were endotracheal instilled LPS (lg/L, 0.2 ml) on the first and 15^th day (no passive smoking on the same days) and they received passive smoking from the 2nd day to 28th day. The rats undertook passive smoking for 3 times. Each time they received 10 cigarettes lasting for 30 minutes and the interval time was 2 hours. Except the adjustment of the dose of LPS to 0.1 ml on the 15th day, the dose of cigarettes to 5 from the 2nd day to the 7th day, other performances in the modification groups are the same as those in the model group. On the 29th day, the pulmonary function, histopathology changes, concentration of TNFa in bronchoalveolar fluid (BALF), success rate and mortality rate were compared among three groups. Results Compared with the rats in normal group, the latter groups rats，airway resistance and concentration of TKFa in BALF were much higher. Conclusion This modified method of passive smoking and endotracheal instillation of LPS for establishment of COPD models in rats is more economic, reliable and worthy of application.

Morris Water Maze Video Behavioral Analysis Using Imagej

FU Peng-cheng，ZHOU Lin，LI Min，YANG Qi-dong

2009, 29(3):  157-161. 

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Objective To analyzed Morris water maze video based on the usage of ImageJ. Methods USB camera and common PC (Intel Pentium 4 CPU 1,70GHz) were used, and it's not necessary to mark the animals. After the procedures of setting scale, removing background and binaryzation with ImageJ, we used MultiTracker，an ImageJ plugin ,which can track the movement of multiple objects，to analyze the results. Result Swimming paths of rats and paths can be successfully analyzed. Conclusion This method based on a free software and low hardware requirements, can get all necessary date of Morris water maze.

Establishment of ELISA Method for Detection of Monkey B Virus

LI Xiao-bo，XIAO Jing，FU Rui，HE Zheng-ming

2009, 29(3):  162-167. 

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Objective To express the glycoproteins of simian B virus, use them to detect the BV antibodies in serum,and search for the better antigens or their combination which have best sensitivity and specificity. Methods Express B virus glycoprotein C，D and G and check their antigenicity by Western Blot after purification and renaturation.The ELISA was established and evaluated. Results The three antigens react well with simian serum which BV antibody is positive. The sensitivity of the antigens is BVgC-ELISA 84.6%, BVgD-ELISA 88.5%，BVgC+gD-ELISA 96.2%, specificity is BVgC-ELISA 93%, BVgD-ELISA 85.7%, BVgC+gD-ELISA 64.3% according to the results of BV-ELISA. Conclusion The three antigens react well with simian serum and can be used as surrogate antigens of BV.

Studies on Differentiation of Porcine Embryonic Germ Cell

KUANG Ling1，FENG Shu-tang2，MU Yu-liang2，LIU Lan2

2009, 29(3):  168-171. 

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Objective Using RA and DMSO to induce porcine embryonic stem cells, they were certificated that could be differentiated into the various kinds cell. Methods Porcine embryonic stem cells were induced to embryonic bodies in culture of free feed or culture of suspension. Embryonic bodies were cultured in the medium with RA or DMSO respectively, to induce embryonic stem cell to differentiate. Results The embryonic bodies would differentiate into neuron-like cells, trophoblast-1 ike cells, morula-like colonies and ES-like colonies when they cultured in the medium with RA and DMSO. Conclusion Porcine EG cells could be differentiated into the various kinds cell. It was testified that porcine EG cells were pluripotency.

Studies on Differentiation of Porcine Embryonic Germ Cell

DING Yi-jing1，WANG Chun-fang1，LI Xiao-tian1，ZHOU Guang-xing2，PAN Hua2

2009, 29(3):  172-176. 

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Objective Using RA and DMSO to induce porcine embryonic stem cells, they were certificated that could be differentiated into the various kinds cell. Methods Porcine embryonic stem cells were induced to embryonic bodies in culture of free feed or culture of suspension. Embryonic bodies were cultured in the medium with RA or DMSO respectively, to induce embryonic stem cell to differentiate. Results The embryonic bodies would differentiate into neuron-like cells, trophoblast-1 ike cells, morula-like colonies and ES-like colonies when they cultured in the medium with RA and DMSO. Conclusion Porcine EG cells could be differentiated into the various kinds cell. It was testified that porcine EG cells were pluripotency.

Effects of DMSA Combined with Vitamin C and L-methioine on Concentration of Lead in Blood and Morphological Changes of Liver Cells in Lead-intoxicated Mice

ZHU Jia-yin1，ZHAO Hui-ling1，GUAN Min-qiang1，WANG Yi-long1，CHEN Tong-ke1，REN Xiao-li1，LOU Wen-wen2，CHEN Xi-wen1

2009, 29(3):  177-180. 

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Objective To investigate the combination effects of dimercap to succinic acid(DMSA)， vitamin C and L-methioine on the ICR mice induced by lead. Methods The mice of the experimental group were exposed to 0.1% lead acetate in drinking water for 6 weeks，which were randomly divided into model control group，low dosage group of DMSA，high dosage group of DMSA and group of low dosage of DMSA combined with vitamin C and L-methioine. Afler 4 weeks of treatment ,the concentration of lead in blood was detected, and the morphological changes of liver cells was observed by Hematoxylin and Eosin Staining. Results The difference of the concentration of lead in blood of model control group was more higher than those of low dosage group of DMSA ,high dosage group of DMSA or group low dosage of DMSA combined with vitamin C and L-methioine (P＜0.01). The difference of the concentration of lead in blood between normal group and model control group was very significant (P＜0.01). The liver cells appeared hydropic degeneration and the effect of combination group was significantly superior to other groups. Conclusion The effect of lead excretion for treating of DMSA combined with vitamin C and L-methioine more better than others.

Relationship between Polymorphisms of PRLR Gene and Reproductive Performance in Tibet Mini-Pigs

ZENG Zhao-zhi1，JIANG Tao1，ZHANG Jin-hong1，GU Wei-wang2

2009, 29(3):  181-184. 

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Objective To obtain the information of AIu I polymorphism distribution of PRLR gene and to study its genetic effect on growth and reproductive performance in Tibet mini-pigs. Methods The litter size and numbers weaning, data of birth weight and weaning weight of piglet from 44 Tibet mini-pig sows were recorded. The AIu I polymorphism of PRLR gene was detected by PCR-RFLP method, and the relationship between the gene polymorphism and the growth and reproductive performance was analyzed by SPSS10.0. Results 3 genotypes of AA、AB、BB were detected in 44 Tibet mini-pigs, the frequency of genotypes was 0.273、0.500、0.227 respectively. Except there was a significant difference in birth weight of latter parity between different genotypes, the other index of growth and reproductive performance between different genolypes had no significance difference. Conclutions There was no significant difference on litter size and numbers of weaning between different genotypes of Tibet mini-pigs.

Evaluation Index System of Preparation Efficacy on Sodium-selenite-induced Cataract in Neonate Rats

CUI Shu-fang，YU Chen-lin，CAI Li-ping，LIU Zhi-xue

2009, 29(3):  185-191. 

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Objective To design the evaluation index system of preparation efficacy on sodium-selenite-induced cataract in neonate rats，on the basis of the animal models’ establishment，combined with the establishment principle of evaluation index system and the preparation principle of animal models of human disease. Methods Through literature research, 26 indicators for evaluating the system for sodium-selenite-induced cataract in neonate rats were collected . According to the research report and the effects of various factors on the preparation of sodium-selenite-induced cataract in neonate rats, coupled with Delphi method and AHP (analytic hierarchy process), we designed the preparation effect evaluation index system and established the animal model. Results Through the evidence, the evaluation index system was confirmed as a good system. Conclution The evaluation index system set up in this study was a clear, simple and easy-to-use system.

Newt's Great Ability of Regeneration

CHEN Xi-ping1，ZHANG Lei2，LAN Cai-juan2，YANG Fang2，Chen Xi-zhe2

2009, 29(3):  207-207. 

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As one of the urodele amphibians, newt can regenerate whole limbs, tails, lens, jaw bones, spinal cord and forebrain of central nervous system, also the part of 20% heart. Newt, regarded as “regeneration king of vertebrates”，has the great ability of multi-organ regeneration. In recent years, although the cellular basis of regeneration has been studied in detail, we know relatively little about the molecular of the process. This review mainly focus on the dedifferentiation process which yields a population of embryonic-like pluripotential cells that will eventually regenerate the lost structure. With summarizing the studies on the basis of cellular and molecular aspects of limb regeneration in the newts, it seems to enlighten us on mammal regenerative biology and even on human regenerative medicine.