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Injection of ethanol into the common bile duct to establish a rat model of biliary atresia

Juntao GE ; Long LI ; Yandong WEI ; Haibin WANG ; Guoliang QIAO ; Zhen ZHANG ; Yao LIU ; Anxiao MING

Acta Laboratorium Animalis Scientia Sinica.2014;(3):50-52. doi:10.3969/j.issn.1005-4847.2014.03.010

Objective To establish a new rat model of biliary atresia by pure ethanol injection into the common bile duct.Methods A catheter was inserted and fixed in the common bile duct in male SD rats .Saline (8 rats) or pure ethanol (16 rats) was injected through the catheter ,respectively, and the biochemical and pathological changes in the rats were examined .Results SD rats in the experimental group were divided into a persistent injury and a restoration of liver dysfunction groups according to pathological and biochemical detection .In the persistent injury group , biochemical impair-ments were significantly higher at 8 weeks after ethanol injection than those in the control group and restoration group .Dis-tinct pathological changes in the liver were observed using HE , SMA, and Masson staining .Conclusions It is a reliable animal model of biliary atresia induced by injection of pure ethanol into the common bile duct in the rat .It will provide a useful tool in future studies of biliary atresia .

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Effect of traditional Chinese medicine composit Guben Yiliu III combined with gemcitabine on human pancreatic cancer xenograft in nude mice

Ju LIU ; Haili QIAN ; Zhijian XU ; Lisheng ZHANG ; Peizhi YE ; Aiping TIAN ; Hongli YANG

Acta Laboratorium Animalis Scientia Sinica.2014;(3):98-102. doi:10.3969/j.issn.1005-4847.2014.03.020

Objective To explore whether the Chinese medicine Guben Yiliu III can improve the effect of gemcit -abine on human pancreatic cancer xenograft in nude mice . Methods Nude mice with transplanted human pancreatic cancer were divided randomly into 4 groups: control group, gemcitabine treatment group , combined ( Guben Yiliu III +gemcitabine) group, and Guben Yiliu III group, 10 mice in each group.The gemcitabine group and combined group were treated with gemcitabine from the 8th day after transplantation in a dose of 100 mg/kg by i.p.injection, twice a week. Guben Yiliu III and combined groups were given the aqueous solution of Guben Yiliu III granules p .o.since the 8th day af-ter transplantation .Result The inhibition rate of transplanted tumor in the three treatment groups were 48.9% in the gemcitabine group , 68.9%in the combined group , and 28.0%in the Guben Yiliu III group .The combined group showed a significantly higher inhibition rate than the gemcitabine group (P<0.05).The gemcitabine group, combined group and Guben Yiliu III group showed a significantly slower growth rate than the control group .However, the combined treatment group showed a pronounced side effect and body weight loss than the other 3 groups .Conclusions The Chinese medicine Guben Yiliu III can improve the inhibitory effect of gemcitabine on nude mice with human pancreatic cancer xenograft in the auxilla of nude mice .

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Electrophysiological study of BDNF gene-modified mesenchymal stem cell transplantation to repair transversely hemisectioned spinal cord injury in rats

Meng JI ; Yuqiu LIU ; Jingjie ZHAO ; Chengwen JIN ; Xin LI ; Zhixin WEI ; Xiaocui WANG

Acta Laboratorium Animalis Scientia Sinica.2014;(3):93-97. doi:10.3969/j.issn.1005-4847.2014.03.019

Objective To study the effects of bone marrow mesenchymal stem cells modified by brain-derived neuro-trophic factor ( BDNF) gene on the repair of spinal cord injury by electrophysiological assay .Methods Thirty healthy Spra-gue-Dawley rats (male and female) were randomly divided into 3 groups:Blank group, 10 rats (removal of the lamina only and exposed spinal dura mater );spinal cord injury (SCI) group,10 rats;and cell transplantation after SCI group , 10 rats. Eight rats of them were selected randomly and detected their SEP and MEP , and evaluated the degree of recovery of motor scores in the rats at 1 d, 7 d, 14 d, 21 d, 30 d, and 60 d.Result Since 4 days after cell transplantation , the process of hind limbs changes was as follows:at the 1-4 days after injury , the injury side hind limb had flaccid paralysis , mopping the floor walk, the movement of contralateral hind limb was gradually recovered from the initial injury , the injury side hind limb had spastic paralysis in 5-9 days after SCI;during 10-14 days, the injury side had a few activities;the contralateral side re-covered to a less normal state;At 15-21 days, activities of the injury side improved obviously , until the 30th day.The activ-ity and muscle tension degree of the injury side recovered most obviously .After 30 days no more obvious improvement was ob-served.Immunohistochemistry showed that the transplanted mesenchymal stem cells , which were induced and labeled firstly , survived at the damage spinal cord , and behavioral observation found that the cell transplantation improved exercise capacity of the rats injured before .Conclusion Bone marrow mesenchymal stem cells modified by BDNF gene can partially promote the recovery of nerve transmission function and nerve regeneration .

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Development of a new coating substrate for human embryonic stem cell culture

Hailin ZHU ; Hui GONG ; Xiangya ZHOU ; Jinliang YANG ; Yuquan WEI ; Huimin CHEN

Acta Laboratorium Animalis Scientia Sinica.2014;(3):88-92. doi:10.3969/j.issn.1005-4847.2014.03.018

Objective To reduce the animal component contamination for human embryonic stem cells ( hESCs ) and to simplify hESCs culture process , we develop a new coating substrate which can support the hESCs growth without dif -ferentiation, and is easy to store and use. Methods Mouse embryonic fibroblasts(MEF)were fixed on the surface of plate by methanol.hESCs were cultured on this new substrate and were passaged every 5 to 6 days.After 10 passages, we checked the cell morphology , alkaline phosphatase expression , embryonic specific markers and the differentiation ability in vitro.Results After 10 passages , the hESCs grew well on this new substrate and maintained the typical hESCs morpholo -gy.Alkaline phosphatase staining was positive .Immunofluorescence staining showed that the expressions of Oct 4, SSEA4, Tra-1-60 were positive .The cells formed embryoid body in vitro .Conclusions This methanol-fixed MEF substrate can support the growth of undifferentiated hESCs .The coating material can be produced in large scale and stored for a long time.It provides a new and relatively easy way to amplify hESCs .

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Comparison of early developmental differences of hair follicles in different skin areas of neonatal mice

Yuchen JIA ; Qi CHEN ; Shaowei LI ; Yuanqiang ZHENG ; Li WANG

Acta Laboratorium Animalis Scientia Sinica.2014;(3):84-87. doi:10.3969/j.issn.1005-4847.2014.03.017

Objective The aim of this study was to observe the growth difference and expression of cytochrome C of skin hair follicles in neonatal mice .Methods The morphology of different skin hair follicles of neonatal mice ( postnatal day 1-9)were observed by HE staining histology and cytochrome C was detected by immunohistochemistry .Results The skin hair follicles in different parts of neonatal mice showed differences not only in morphology but also in developmental pe -riods.Hair follicle growth in the back and tail skin had a nonlinear and growing period .After the nonlinear and growing pe-riod they began to grow rapidly .The tail development was slightly slower than that on the back .The hair follicles of vibris-sae were very special , and started to develop without a stable period .Conclusions The results of morphological observa-tion and cytochrome C immunohistochemistry demonstrate that differences exist in the hair follicle morphology and develop -mental times in the skin of different parts of the body in neonatal mice .

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26 polymorphic microsatellite markers screened from the genome of guinea pigs

Diwen LIU ; Weiwei YANG ; Baojin WU

Acta Laboratorium Animalis Scientia Sinica.2014;(3):78-83. doi:10.3969/j.issn.1005-4847.2014.03.016

Objective To screen microsatellite DNA markers from genome of guinea pigs for further genetic quality control and gene-mapping of this species .Methods Microsatellite sequences were obtained by magnetic bead enrichment and genome database screening , and candidate loci were chosen to design primers .Thereafter , genomic DNA of 5 different guinea pig strains were employed to select polymorphic microsatellite DNA markers based on PCR amplification results .Re-sults A total of 304 microsatellite sequences were analyzed by magnetic bead enrichment and 125 primers were designed . One polymorphic microsatellite DNA marker and 17 specific sites ( no polymorphic was found ) were determined .By gene-mapping , 292 microsatellite sequences were obtained and 178 primers were analyzed , totally 25 polymorphic microsatellite DNA markers and 28 specific sites ( without polymorphics ) were discovered .Conclusions We obtained 26 polymorphic microsatellite DNA markers and 45 potential markers in guinea pigs , and these may lay a foundation for application of mic-rosatellite DNA markers in genetic quality control and gene-mapping of guinea pigs .

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Application of reverse dot blotting for detection of Tyzzer’ s organism

Tingting ZHAO ; Lei YAN ; Liwen WEI ; Li WEI ; Huijuan WANG ; Guoqi LAI ; Yi TAN

Acta Laboratorium Animalis Scientia Sinica.2014;(3):72-77. doi:10.3969/j.issn.1005-4847.2014.03.015

Objective To establish a simple, stable, specific and sensitive method for detection of Tyzzer ’ s or-ganism by reverse dot blotting ( RDB) .Methods Primers and specific probes were designed according to the conservative sequence of Tyzzer 16S rDNA.The forward primer was labeled with biotin .The reverse dot blotting method was established followed by PCR amplification .The specificity and sensitivity of this method were determined .Next, 41 mice and 38 rats were examined by RDB , ELISA and IFA .Results The RDB method showed a high specificity , and in the testing of the 79 laboratory animals , its limit of detection was 4.5 ng/μL.Compared the results of ELISA and IFA , its consistence with ELISA was 100%and the positive rate was 7.59%(6/79), the consistence with IFA was 92.4%(73/79), and the posi-tive rate was 0%.Conclusions An accurate, sensitive and specific method in combination with PCR and RDB in detection of Tyzzer’ s organism is established in this study .

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Effects of living environment conditions on the blood hormone levels and psychological behavior in Chinese tree shrews

Farong YU ; Qingyun CHANG ; Xiuzhen LIAN ; Zhennan ZHANG ; Mingren XIE ; Denglou LI

Acta Laboratorium Animalis Scientia Sinica.2014;(3):62-66. doi:10.3969/j.issn.1005-4847.2014.03.013

Objective To study the effects of living environment conditions and animal-animal interaction on the blood hormone levels and psychological behavior in Chinese tree shrews .Methods Chinese tree shrews were raised in ca-ges of different space sizes or were administered reserpine for 15, 30, 60, 120, 180 days, respectively.Then the animals were anesthetized by ether inhalation , and blood samples were taken from the heart to detect the levels of blood testosterone (T), estradiol (E2), endothelin (ET), adrenaline (A) and noradrenaline (NA) by radioimmunoassay (RIA).Results 1.Chinese tree shrews were bred in large cage (D1group) or small cage (X1 group) for 15, 30, 60, 120, 180 days, re-spectively.Compared with the animals bred in the large cage (D1 group), the level of blood testosterone (T) was signifi-cantly reduced (P<0.01), and the levels of adrenaline , noradrenaline and endothelin were significantly increased in the small cage group (P<0.01 for all).2.The animals raised in small cages in close neighborhood with large cages for 15, 30, 60, 120, 180 days, respectively.The levels of testosterone, adrenaline and noradrenaline in the large cage group (X2 group) were significantly higher than those of the small cage group (X1 group) (P<0.01 for all).3.The adrenaline and noradrenaline levels were significantly lower in all the reserpine groups (P<0.01 for all).4.The animals bred in small cages (X1 group) and in small cages in close neighborhood with large cages showed sudden loss , reduced appetite, testicu-lar atrophy , penile prolapse and stress symptoms .The animals of the reserpine groups appeared gentle temperament , signif-icantly reduced activity and reduced appetite .However , after stopping the reserpine administration and feeding them in large cages , the animals gradually returned to normal behavior .Conclusion Both animals living environment conditions and animal-animal interaction may cause changes of blood hormone levels and psychological behavior in Chinese tree shrews.

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Effect of CD82 on the expression of integrin αV,β3, E-cadherin andβ-catenin in uterine epithelial cells in pregnant mice

Xiaowei WEI ; Dongmei TAN ; Qian ZHANG ; Tongchuan HE ; Yi TAN

Acta Laboratorium Animalis Scientia Sinica.2014;(3):57-61. doi:10.3969/j.issn.1005-4847.2014.03.012

Objective Uterine epithelial cells were isolated from pregnant mice and cultured in vitro , and exam-ined the effect of CD82 on the expression of integrin αV,β3, E-cadherin and β-catenin in the cells.Methods The uter-ine epithelial cells were primarily isolated from pregnant mouse uterus .The recombinant adenovirus containing mouse CD 82 gene which had been constructed in our lab infected the uterine epithelial cells .Immunocytochemistry was used to detect the protein expressions of integrin αV,β3, E-cadherin and β-catenin in the uterine epithelial cells , which were infected with CD82 adenovirus or not .Results 1.The purity of primary cultured cells was (93.2 ±0.6)%.2.The transfection efficiency of CD82 recombinant adenovirus was ( 92 ±4.5 )%.The adenoviral particles carrying CD 82 gene indeed ex-pressed CD82 gene and protein in the primary uterine epithelial cells after 24 hours or 48 hours.3.The uterine epithelial cells of pregnant mice on d4 expressed integrin αV, β3, E-cadherin and β-catenin.4.In contrast to the control group, when CD82 adenovirus infected cells , the uterine epithelial cells of pregnant mice on d 4 increased the expression of integrinαV,β3 and β-catenin protein, had no significant changes of E-cadherin.Conclusions CD82 may have effect on the ex-pression of integrin αV,β3 and β-catenin in mouse uterine epithelial cells before implantation .

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Bioinformatics analysis of mice Agouti gene polymorphism

Xiaodan HE ; Ying ZHAO ; Liya ZHAO ; Junhua XIAO ; Yuxun ZHOU ; Kai LI

Acta Laboratorium Animalis Scientia Sinica.2014;(3):7-14. doi:10.3969/j.issn.1005-4847.2014.03.002

Purpose Polymorphisms of candidate gene Agouti was analyzed in order to reveal the molecular mech -anisms of coat color difference in chromosome engineering mice .Methods Firstly, differences of mouse coat color was detected by color measurement spectrophotometer .Then, candidate gene Agouti was found by whole genome scanning based on DNA chip.Finally, cDNA and amino acid sequence polymorphisms were analyzed , as well as the influence of protein properties and function after mutation was predicted by bioinformatics software .Results There are five SNPs in the Agou-ti cDNA sequences , resulting in three missense mutations in the amino acid sequence of Agouti signaling protein .Bioinfor-matics analysis revealed that one βsheet deletion in the secondary structure of the mutant protein , as well as tertiary struc-ture changed , leading to decrease of binding ability .Conclusion A novel missense mutation is found in candidate Agouti gene.It plays critical role in receptor binding activity , and may reflect on mice coat color changing from light gray to dark gray eventually .

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