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Establishment of a Rat Model of Ulcerative Colitis by Immunological Sensitization Combined with Local Acetic Acid Irritation

Yuhong WU ; Hailong LI ; Yongqiang DUAN ; Jianjun WU ; Hongwei WANG ; Yanming WEI

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):65-68,彩5.

Objective To establish a rat model of ulcerative colitis by immunological sensitization combined with local acetic acid irritation.Methods Thirty healthy 2-3-month-old Wistar rats were randomly divided into 3 groups:normal control group,model group and SASP-treated model group(n=10 rats per group).The ulcerative colitis in the model group and positive group was developed by immunological sensitization with rabbit colon mucosal proteins and local irritation with 5%acetic acid.The rats of the model and positive groups were fed with sulfasalazine,and rats of the control group fed with normal saline.The rats of positive control group were treated with sulfasalazine.Two weeks later,all rats were killed and colon mucosal and blood samples were collected.The concentrations of NO,NOS,MDA,SOD and GSH-Px in the colonic mucosal homogenate were measured by biochemistry,the contents of IL-4 and TNF-α were measured by RIA,and the serum concentration of IFN-γ was measured by ELISA.Results Compared with the normal control group,the concentration of MDA,NO,NOS in the colonic mucosa was increased,and SOD and GSH-Px levels significantly decreased in the model group.Compared with the model group,the concentrations of MDA,NO,and NOS in the colonic mucosal tissue were decreased,and the levels of SOD and GSH-Px decreased in the SASP-treated model group.The contents of IL-4 in the serum and colonic mucosal homogenate of model group were significantly lower, and the contents of TNF-α in the serum and colonic mucosal homogenate were significantly increased than those of the normal control group. The IL-4 levels in the serum and colonic mucosal homogenate was increased, and that of TNF and IFN-γ significantly decreased in the SASP-treated model group, compared with those of the normal control group.Conclusion The rat model of ulcerative colitis established by immunological sensitization combined with local acetic acid irritation, as described in the present study, can better simulate the characteristic chronic alterations in human ulcerative colitis,and may better serve studies and evaluation of treatment of this disease.

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Establishment and Evaluation of a Rat Model of Peritoneal Bacterial Infection after Liver Transplantation

Long DING ; Yu YANG ; Jiahong DONG

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):69-73,彩7.

Objective To establish a rat model of peritoneal bacterial infection after liver transplantation.Methods To construct a dark Agouti rat-to-Lewis(DA-to-LEW) rat model of liver transplantation.Peritoneal bacterial infection in the rats was induced by intraperitoneal injection of bacterial suspension.The liver function,blood gas,blood cell count and other indicators of the rat models were detected.Results There was a high mortality rate in rats with bacterial injection at day 5 after liver transplantation,therefore unfavorable for the following study.It waft better to inject the bacteria in an amount of 5×10~5 cfu/mL at day 3 after liver transplantation.The cumulative 7-day survival rate of those rats after infection reached up to 37.5%.The infection became increasingly severe,the general conditions were worsening,the rectal temperature was rising,the WBC count was increased,the pH was decreased,liver dysfunction was progressively increased,and metabolic acidosis occurred in the rats.Liver parenchymal damage was more pronounced than that of bile ductal injuries,and the rats died one after another at about 5 days after infection.Pathological examination of multiple organs showed that the main cause of death of the rats was liver damage,without accompanying lung and kidney damages.Conclusion The results of this study suggest that it is a successful method to establish a rat model of peritoneal bacterial infection after liver transplantation,and this model can be used in related experimental researches.

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Bone Marrow Mesenchymal Stem Cell Transplantation in the Repair of Rat Spinal Cord Hemisection Injury

Jinsheng WU ; Aiping DONG ; Xiaocui WANG ; Zhixin WEI ; Weiguang LIU ; Zhimin LUAN ; Zengjun ZHU

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):-.

objective To investigate the differentiation of bone marrow-derived mesenchymal stem cells into neurons and transplantation of the stem ceils to repair rat hemisection spinal cord injury.Methods Adherent culture was used to isolate and culture rat bone marrow mesenchymal stem cells(MSCs).The rat spinal cord homogenate supernatant was used to induce neural differentiation of the 3rd generation ceils.The nature of ceil differentiation was identified by immunohistochemistry.The rat model of hemisection spinal cord injury was prepared and BrdU was locally injected to label the induced neurons.The distribution of living cells in the injuried spinal cord was observed at 5 weeks after cell transplantation.Results MSCs were spindle and polygonal,with 1-2 nucleoli seen under the inverted microscope.After induction with spinal cord homogenate supernatant there were a number of slender cytoplasmic projections forming interwined network and showing nestin expression,therefore,indicating the neuronal nature.MSCs at 5 weeks after transplantation into the spinal cord injury were surviving and their expression of MAP-2,NF,GFAP was significantly higher than that in the control rats(P<0.05).The rat motor function was improved than before transplantation.Conclusion MSCs induced by spinal cord homogenate supernatant can be transplanted into hemisection spinal cord injury and improve the motor function of the injuries lesions.

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Expression of Nerve Growth Factor (NGF) in Mouse Testes at Different Developmental Stages

Yongting HU ; Wenhua CHEN ; Ying ZHANG ; Xiangting NIE

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):28-32,封3.

Objective To study the location and the expression of nerve growth factor(NGF)in mouse testes at different developmental stages.Methods The testes were taken out from mice at different developmental stages,respectively.Parts of the testis tissue were used to extract total RNA and real-time PCR was used to detect the expression of NGF.The rest tissues were fixed and embedded in paraffin.SABC immunohistochemical staining was used to observe the location of NGF in the testes. Results Reat-time PCR results showed that NGF mRNA could be detected in the testes of mouse at postnatal 1 st week and its expression level increased with the development,and reached the highest point at 3rd week.After 5 weeks it lowered down to a stable level.Immunohistochemistry showed that NGF expressed in the mouse testes at all developmental stages.At postnatal 1 st week immunopositive reaction of NGF was detected mainly in sustentacular cells and the spermatogonia also showed positive staining.NGF positive staining in the testes was observed in interstitial cells,spermatogenetic cells,sustentacular cells and Leydig cells at 3rd week.After the postnatal 5th week,NGF-positive immunostaining was also detected in intersitial cells and spermatogenetic cells,but the intensity of reaction was weaker than that at 1 st and 3rd weeks.Conclusion The expression levels of NGF in mouse testes show regular changes and the expression profile and location of NGF in the testes are different at different developmental stages.

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Morphological and Morphometric Study on the Effect of Soybean Isoflavone on Hyperplastic Prostate in Rats

Guofeng REN ; Quan YUAN ; Aiqing YANG ; Ling TANG ; Yiming HUANG

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):24-27,彩6.

Objective To investigate the effect of soybean isoflavone on histology and uhrastructure of benign prostatic hyperplasia in rats.Methods Male SD rats were injected subcutaneously testosterone propionate for 28 d to induce prostatic hyperplasia,and the rats were divided into 5 groups:control group,model group,and 3 test groups with SI in a dose of 60 mg/(kg·d),120 mg/(kg·d) and 240 mg/(kg·d),respectively.The wet prostate weight,prostatic index,morphological,uhrastmetural and morphometrie changes of the prostatic shndular and interstitial tissues were observed.Results The prostate wet weight,prostatic index and prostatic volumes in all dose groups were significantly lower than those in the models.In comparison with the model group,height of prostatic epithelial cells,glandular average diameters,volumes and surface areas in unit volume,as well as glandular circumferences,glandular relative total volumes and interstitial relative total volumes were all significantly decreased.Glandular counts,density,ratio of glandular surface area to volume,and glandular average curvature were all increased.Conclusions Soybean isoflavone can inhibit prostatic hyperplasia in rats.

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Expression and Significance of Cyclin A1 mRNA in the Skin of Wild-Type Mice

Xia ZHU ; Hongquan WEI ; Yuanhong LI ; Yan WU ; Chen ZHAO ; Xinghua GAO ; Hongduo CHEN

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):21-23,彩5.

Objective To study the expression and significance of cyclin A1 in the skin of wild-type mice at RNA level.Methods Thirty 6-12-week old wild-type Kunming mice(15 male and 15 female)were included in this study.In situ hybridization was used to detect the expression of cyclin A1 mRNA in the skin of head and neck of the mice.The skin treated without probe was regarded as negative control and the testis of male mice was taken as positive contr01.Results The expression of cyclin A1 mRNA was found in sebaceous glands of 25 mice and in epidermis of 12 mice.Strong positive staining of sebaceous glands was seen in 50% and positive staining in 33.3% of sections,whereas strong positive and positive staining of epidermis was seen in 13.3% and 20% of sections.respectively.The positive rate of sebaceous glands was 83.3%,much higher than that of epidermis(33.3%).Conclusions There is a quite high expression of eyelin A1 mRNA in the skin sebaceous gland and epidermis of head and neck of wild-type mice.Especially a strong expression is in the sebaceous glands.It indicates that cyclin A1 mRNA may play a certain role in the physiological function of sebaceous glands and epidermis of the skin.

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Analysis of Factors Affecting the Efficiency of Establishment of Mouse Parthenogenetic Embryonic Stem Cells

Zhisheng CHEN ; Haojia WU ; Lin LIU

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):52-55,彩6.

Objective To study the factor affecting the efficiency of establishment of parthenogenetic stem cells (PESCs)in mice. Methods PESC lines were derived from parthenogenetically activated blastocyst inner cell mass of different mice strains and cultured in different systems.Results The efficiency of establishing the pESC showed no significant difference in hybrid and inbred lines.But the efficiency was increased in culture systems added with ERK inhibitor or knockout serum replacement(KSR).Conclusion The efficiency of establishment of pESC was not directly related with the genetic background of mice but it was closely related to the culture systems.

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Establishment and CT Diagnosis of Orthotopic Rat Model of Bladder Cancer

Xiaoying TANG ; Chunwu PAN ; Jun SUN ; Wei HUANG ; Huafeng WANG ; Zhoujan SHEN

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):6-8,彩2.

Objective To establish an orthotopic rat model of bladder cancer induced by N-methyl.nitrosourea (MNU)and to evaluate the diagnostic value of CT(computed tomography)scanning in this experimental model.Methods 50 SD rats were randomly divided into the control group(group A)with 15 rats and experimental group(group B)with 35 rats.Each rat of group B was treated with 2 mg MNU per dose every other week,totally 4 doses,by per urethra administration.Meanwhile,each rat of group A was treated with normal saline.Then,at the 14th week,all the rats were evaluated by CT scanning and pathological examination.Results Abnormal changes were detected in each of the 28 rats in group B by CT scanning,and manifested as local mass,thickening bladder wall accompanied with heterogeneous density.Bladder cancer was diagnosed by pathology.However,no bladder tumor was detected by CT scanning and pathologicalexamination in group A.Conclusion A rat model of orthotopic bladder cancer can be established by per urethra administration of MNU.CT scanning is a reliable diagnostic technique concerning this model.Furthermore,this technique can render US a more suitable rat model for further experimental studies on chemotherapy and radiotherapy of bladder Cancer.

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Comparison of Experimental Autoimmune Encephalomyelitis in Mice Induced with Myelin Oligodendrocyte Glycoprotein at Different Doses

Yang MU ; Hui ZHAO ; Yan LIU ; Lei WANG

Acta Laboratorium Animalis Scientia Sinica.2010;18(1):1-5,彩1.

Objective To compare the condition of illness and pathological characteristics of experimental autoimmune encephalomyelitis (EAE)in C57 BL/6 mouse models induced by myelin oligodendrocyte glycoprotein 35-55 (MOG35-55)at different doses,and provide a reliable animal model for further study of multiple sclerosis(MS).Methods Male SPF-grade C57 BL/6 mice were divided randomly into four groups:normal group and three EAE model groups (MOG35-55 high-dose,middle-dose and low-dose model groups).200,100,50μg MOG35-55/mice were mixed with complete Freund's adjuvant(CFA),respectively,to prepare complete antigen in different concentrations.The mice were anesthetized and injected s.c.over flanks with the complete antigen and injected i.P.with pertussis toxin to establish immunization-induced C57BL/6 mouse-model of EAE.The mice of the normal group were injected with normal saline instead.Since the day of immunization,the incidence,body weight and neurological score of the mice were observed.The mice of different neurological scores in different periods were anesthetized and perfused with saline and followed by 4% paraformaldehyde.The brain and spinal cord of the mice were removed and fixed in the same fixative solution.The brains and spinal cords of the mice were examined by histopathology with hematoxylin-eosin(HE) staining.The mice on the 40th day were sacrificed and perfused with 2% paraformaldehyde and 2% glutaraldehyde, 1 mm~3 pieces of cerebral white matter and intumescentia lumbalis of the spinal cord were taken and ultrathin sections were prepared according to conventional techniques for electron microscopy. Results All the MOG_(35-55) in three different doses induced mouse models of EAE. The disease was with an incidence rate of 100% and a chronic monophasic course. The body weight of the mice in the three groups decreased obviously compared with those in the normal group. The maximum value of neurological score was 1.33,2.25 and 2.50 in the mice of high-, middle-and low-dose groups, respectively. The major histopathological changes observed in the brain and spinal cord of the EAE mice were different degrees of inflammatory cell infiltration around small vessels showing sleeve-like changes, dcmyelination and neuronal karyopyknosis in the acute and remission stages. The main site of the brain inflammation was in white matter around encephalocoele, and also in the DG and CA zones of hippocampus. The spinal cord inflammation was most severe in the lumbosacral region. The above mentioned pathological changes in the low-dose group were more prominent than those in the middle-dose and high-dose groups. The major ultrastructural changes were scattered around encephalocoele, interstitial edema, especially around small blood vessels, and swollen mitochondria with damaged cristae, and some karyopyknosis in vascular endothelial cells. Some tight junctions were blurred. Some dispersed lymphocytes and mononuclear cells were seen in the perivascular space. In lumbar intumescentia of the spinal cord, there were some myelin figures in the white matter myelin sheath. Some of them showed demyelization and structurtal fusion. The cytoplasmic organelles of axons were considerably reduced or even disappeared. The vascular basement membrane showed an increased thickness and focal necrosis in some areas. Conclusion The mouse models of immune-induced EAE are successfully established with MOG_(35-55), especially that induced with MOG in a dose of 50 μg. This mouse model is stable, with a high incidence and low mortality rate, and can be applied for EAE research in the future.

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Comparison of Three Procedures in the Establishment of Rat Models of Focal Cerebral Ischemia

Hao ZHAO ; Yongning LI ; Renzhi WANG ; Xinjie BAO ; Yingjie ZHAO ; Guilin LI ; Junji WEI ; Ming FENG ; Yanguo KONG

Acta Laboratorium Animalis Scientia Sinica.2009;17(6):432-436,illust 2.

Objective To compare and evaluate the efficacy of three operating procedures to produce permanent rat models of cerebral ischemia, including mortality, neurological evaluation score, infarction volume of ischemia and time consumed in the operation. Methods The rats were randomly divided into three groups. Group 1: The common carotid artery (CCA) and external carotid artery (ECA) were ligated during the operation and ICA was clipped temporarily by artery clamp. Group 2: The CCA and ECA were ligated and the superior thyroid artery and pterygopalatine artery were exposed but not ligated. Silk suture was utilized to hang the internal carotid artery (ICA). Plastic suture was inserted into ICA and stepped over the initiation point of pterygopalatine artery under microscopic observation. Group 3: The CCA and ECA were ligated and ICA was hanged with a silk suture, and the pterygopalatine artery was not exposed, but during the period of inserting plastic suture, the proximal part of the suture was pressed to make the suture's round distal end elevated, and then pass the initial point of pterygopalatine artery. Mortality, neurological score, volume of ischemic infarction and operation time consumed of the three groups were compared after the operation. Results The procedure to prepare the model was most efficient in the group 3, taking only 17.5 min to complete, significantly less than that in the group 1 (50 min) and group 2 (40 min), (P<0.05), and with a lower mortality and more steady neurological evaluation score and infarction volume. Conclusion The use of the third operating method can shorten the operation time and improve the efficacy of operation. Using this method, more consistent and repetitive focal cerebral ischemia models can be produced effectively, and meet the demands of clinical trials.

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