Table of Content

25 February 2011, Volume 31 Issue 1

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Application of Synchrotron Radiation Microangiography on Rat Middle Cerebral Artery Occlusion Models

YUAN Fa-lei, GUAN Yong-jing, XIAO Ti-qiao, XIE Hong-lan, YANG Guo-yuan, WANG Yong-ting

2011, 31(1):  11-15.  DOI: 10.3969/j.issn.1674-5817.2011.01.003

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Objective To apply synchrotron radiation microangiography on the study of rat middle cerebral artery occlusion (MCAO) models. Method Twenty Sprague-Dawley rats weighing 300-350 g were divided into two groups: 4-0 suture group and 4-0 silicone-coated suture group. Immediately after MCAO surgery, synchrotron radiation angiography was conducted. Result The plain 4-0 nylon suture failed to completely occlude the middle cerebral artery (MCA) of the rat. There was still blood supply from anterior cerebral artery (ACA) and interior carotid artery (ICA). The silicone-coated 4-0 nylon suture successfully induced rat MCAO with 10% rats had fenestrated MCAs (2/20).The variants in these rats were occluded when the suture depth was 17 mm. Conclusion The silicone-coated 4-0 nylone suture successfully induced MCAO in rats weighing from 300 to 350 g, and the plain 4-0 suture was not able to induce complete ischemia in these rats. Synchrotron radiation microagiography is useful for studying rat MCAO models and evaluating rat cerebral abnormalities.



Establishment of Rat Hepatic Blood Inflow Occlusion Without Hemihepatic Artery Control Model

JIN Shan, DAI Chao-liu, YANG Yun-tian

2011, 31(1):  16-19.  DOI: 10.3969/j.issn.1674-5817.2011.01.004

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Objective To establish the model of rat hepatic blood inflow occlusion without hemihepatic artery control. Methods Animal models were established by three surgical pathways. Pringle maneuver (PR), hemihepatic vascular occlusion (HH) and hepatic blood inflow occlusion without hemihepatic artery control (WH). The operation success rate, rat mortality , rat survival rate, liver pathological changes with 45 minutes hepatic ischemia followed by 24 hour reperfusion were observed. Results Thirty-three Wistar rats were operated and in each group 8 rats had been operated successfully. Operation success rate were respectively 88.9% (8/9) in group PR, 57.1% (8/14) in group HH, 80.8% (8/10) in group WH, the rat survival rate in group PR was only 37.5% (3/8) (hepatocyte spotty necrosis, ballon degeneration and acidophilic necrosis were observed), it was 100% (8/8) in group HH, and 87.5 % (7/8) in group WH. Mild liver pathological changes were obser ued in group HH and WH compared to group PR. Conclusion Hepatic blood inflow occlusion without hemihepatic artery control is a more simple and safe surgical pathway to establish rat hepatic bloodinflow occlusion model.



Culture of Newborn Rhesus Monkey Liver Epithelial Progenitor Cells in Chemical Defined Serum-free Medium

SUN Ai-jing, Huang Di-lai, RU Guo-mei, JIN Li-fang

2011, 31(1):  20-27.  DOI: 10.3969/j.issn.1674-5817.2011.01.005

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Objective to establish a chemical defined serum-free culture system for rhesus monkey hepatic progenitor cells. Methods a rhesus monkey liver polygon cell colony was derived from a small piece of newborn livers using chemical defined serum-free medium. Follow by isolated and cultured in different extracellular matrix. And the identification of the cells phenotype and dual-differentiation potential for functional hepatocytes and biliary duct cells in vitro were carry out using immunofluorescence cell staining, Periodic Acid-Shiff Staining, RT-PCR, indocyanine green (ICG) uptake staining method. Result Two types of cell colony were generated: elongated spindle cells and polygonal epithelial cells. Elongated spindle cells were expressed as vimentin and brachyury, and they were disappeared within 5 days in our cultures. The remaining consisted of small polygonal epithelial cells that expressed cytokeratin 7 (CK7), CK8, CK18, nestin, CD49f and E-cad, the markers of hepatic stem cells, but were negative for 〈-fetoprotein, albumin and CK19. They can proliferate and be passaged, if on laminin or rat tail collagen gel, to initiate colonies. When cultured with dexamethasone and oncostatin M, the expression of mature hepatocyte markers, such as 〈-1-antitrypsin, intracytoplasmic glycogen storage, indocyanine green uptake and lipid droplet generation, were induced in differentiated cells. If transferred onto mouse embryonic fibroblasts feeders, they gave rise to CK19 positive cholangiocytes with formation of doughnut-like structure. Conclusion we isolated a polygon epithelial cell with characteristics of live progenitor cells from newborn rhesus monkey live tissue using a chemical defined serum-free culture system, and. may serve as an in vitro model of primate liver development and may facilitate further preclinical research.



Observation on Dynamic Histopathological Changes in Bladder Tumor Model Induced by N-methyl-N-nitrosourea in Rats

GU Wei, XU Yao-ting, XIE Min, LI Du-jian, HUANG Ru-qiang, XU Xiao-wen

2011, 31(1):  28-32.  DOI: 10.3969/j.issn.1674-5817.2011.01.006

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Objective To establish the bladder tumor model induced by N-methyl-N-nitrosourea in rats and to observe the dynamic histopathological changes of the bladder mucosa. Methods A bi-weekly doses of 2 mg MNU each time for 8 weeks intravesicular administration were given. The changes of the bladder mucosa were pathologically examinated at 3rd, 5th, 7th, 9th, 12th and 14th week. Results Untypical hyperplasia could be observed after 3 weeks of administration, the carcinoma in situ occurred at 7 weeks, distinctive tumor was found at 9 weeks, bladder papilloma or invasive carcinoma was observed at 12-14 weeks. The rate of carcinogenicity was 100%(6/6) after 9 weeks. The histological changes and the pathologic characters were very similar with human bladder tumor. Conclusion The method to induce bladder tumor by intravesicular administration of MNU is convenient and reliable. The carcinogenic process includes epithelial hyperplasia, papilloma and carcinomatous changes. It is an ideal animal model.



Establishment of Subcutaneous and Orthotopic Transplant Tumor Model of Hepatocellar Carcinoma Cell Line with High Metastatic Potential in Nude Rat

PENG Xiu-hua, SHEN Yan, XU Chun-hua, ZHOU Wen-jiang

2011, 31(1):  33-37.  DOI: 10.3969/j.issn.1674-5817.2011.01.007

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Objective To study the feasibility of the establishment of subcutaneous and orthotopic transplantation tumor model of hepatocellar carcinoma cell line with highly metastatic potential (HCCLM3) in nude rat, and its tumor biological characteristics. Methods HCCLM3 cell line was inoculated subcutaneously in nude rat to develop growing tumor. Orthotopic implant model was established by suturing histologically intact human tumor tissue on the liver of nude rat. Body weight of nude rat and diameter of tumor were measured weekly. The animals were then killed 8 weeks post transplantation, the biological characteristics of transplant tumor was observed by gross examination and microscopic examination. Results In subcutaneous transplant model, tumor formation rate was 100% and no lung metastasis. In orthotopic transplant model, tumor formation rate was 90%, metastasis rate of lung was 70%, the rate of mass ascites was 50% and the natural extinctive rate was 0%. Conclusion The orthotopic transplantation tumor model in nude rat is an ideal model for studying the metastatic mechanism and screening anti- tumor drugs for liver cancer, in view of its high successful rate and high spontaneous metastatic rate with no natural extinction.



Efficient Repeated Low-dose Intravaginal Infection with SHIV1157ipd3N4 in Rhesus Monkey

CONG Zhe, JIANG Hong, JIN Guang, WANG Wei, GAO Hong, YAO Nan, TAO Zhen, CHEN Ting, YANG Zhi-wei, Sylvain. Fleury, Anick Chalifour, Ruth. M.Ruprecht, WEI Qiang, QIN Chuan

2011, 31(1):  38-42.  DOI: 10.3969/j.issn.1674-5817.2011.01.008

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Objective To establish a CCR5-specific chimeric simian/human immunodeficiency virus, SHIV1157ipd3N4 repeated low-dose intravaginal infection model with rhesus monkeys. This new chimeric model could hopefully be more efficient model in AIDS vaccine development. Methods Six female Chinese rhesus monkeys were enrolled in this study. All animals were intra-vaginally challenged 11 times with 1 ml of a phosphate-buffer viral solution containing the heterologous SHIV1157ipd3N4, with 20 TCID₅₀ for the first 3 challenges and 10 TCID₅₀ for the remaining 8 challenges. Each challenge was accomplished at the interval of 4-7 days. Whole blood was collected and plasma virus was quantified by real-time SYBR Green RT-PCR and T cell subsets were determined by flow cytometry analysis. Results The systematic infection of six rhesus monkeys was established successfully after 11 challenges. The peaks of viral loads were between 10⁶copies/ml to 10⁸copies/ml.Conclusion The repeated low-dose technique was used for establishing a SHIV1157ipd3N4 infection model of Chinese rhesus monkey. This model aliking sexual route would be very useful for HIV-1 subtype C vaccine and pathogenesis studies.



Evaluation of Methods to Detect Insulin Sensitivity in High-fat Diet Rats

LI Yun, DU Jian, PEI Li-na, LUO Hong-fei, LIU Jia, GUO Ning-ning, ZHANG Ke-ying

2011, 31(1):  48-51.  DOI: 10.3969/j.issn.1674-5817.2010.06.010

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Objective To compare several frequently used surrogate indexes of insulin sensitivity and the euglycemic-hyperinsulinemic clamp experiments in high-fat diet rats. Methods Thirty male Wistar rats were randomly divided into two groups with a regular (NC group, n=15) or high-fat diet (HF group, n=15) for 10 weeks.The euglycemic-hyperinsulinemic clamp experiments,the insulin tolerance tests as well as the glucose tolerance tests were performed and the correlations of the euglycemic-hyperinsulinemic clamp experiments with different types of IR indexes were conducted. Result The glucose infusion rate [GIR (0.90±0.15 vs 4.97±0.68)mg·kg^-1 ·min^-1, P<0.01and KITT (24.41±6.77 vs 48.61±4.71, P<0.01) in HF group decreased significantly. The insulin tolerance tests correlated with the euglycemic-hyperinsulinemic clamp experiments significantly (r=0.911, P<0.01). Conclusion This study demonstrates that the insulin tolerance tests correlated with the euglycemic-hyperinsulinemic clamp experiments significantly.



The Effect of Six Kinds of Traditional Chinese Herbal Composing Prescriptions on Cell Proliferation and Melanogenesis in Cultured B16 Melanoma Cells

ZHANG Jian, WU Wei, WAN Yu-hua, ZHANG Rong-wen, LIU Dan-dan, MA Rui, GU Wei-wang

2011, 31(1):  52-56.  DOI: 10.3969/j.issn.1674-5817.2011.01.011

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Objective To study the effects of six ethanol extracts and aqueous extracts from traditional Chinese herbal composing prescriptions on the cell proliferation and melanogenesis in cultured B16 melanoma cells. Methods The cell proliferation were measured by MTT method, and NaOH assay was to determine melanin synthesis. Results The aqueous extracts of composition 1,2,3 and alcohol extracts of composition 2,4,6 had significantly promoted cell proliferation (P<0.05); aqueous extract with the highest rate of cell proliferation was composing prescription 2 (0.625 mg/ml), the proliferation rate was 22.60%; ethanol extract with the highest rate of cell proliferation was composition 4 (0.078 mg/ml), the proliferation rate was 11.60%. The aqueous extract of composing prescriptions 1,2,4,5,6 and alcohol extracts of the six kinds of composing prescriptions promoted obviously the synthesis of melanin (P<0.05). Aqueous extract with the highest rate of melanin synthesis was composing prescription 1 (0.313 mg/ml), the growth rate is 75.38%; alcohol extract with the highest rate of melanin synthesis was composing prescription 1 (1.25 mg/ml), the growth rate was 88.01%. Conclusion The positive effect of each composing prescription on the proliferation of mice’s B16 melanoma cells and melanin synthesis is various due to the composing of the prescription.

Primary Exploration of Laboratory Animal Dynamic Information Management and Financial Information Management Integration

SUN Qi-hua, WANG Jing, GU Jing, YIN Yin, ZHOU Hui-ying, XUE Zhi-mou

2011, 31(1):  69-72.  DOI: 10.3969/j.issn.1674-5817.2011.01.015

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Laboratory animal is very important for biomedicine industry. However, the commercialization, standardization and socialization of laboratory animal are not enough due to its specialty and the management mode in production. With the help of the laboratory animal dynamic information management and financial information management integrated system, problems in information management and financial accounting would be solved properly, which could facilitate the laboratory animal supply chain establishment and cost control. The development of laboratory animal in college would be accelerated.



Perspective Review of Animal Models for Osteoporosis Research

QIAO Wei-wei, ZHAO Xian-zhe

2011, 31(1):  73-78.  DOI: 10.3969/j.issn.1674-5817.2011.01.016

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Osteoporosis is an important systemic disorder, impairing mainly the health and life quality of the elderly, with a diverse multifactorial etiology. The proper animal models of osteoporosis lay the foundation of sophisticated studies on this disease. Here, we review some typical animal models of osteoporosi and summarize the drawbacks from the previous using experiences and pose the corresponding improvement of these animal models in actual application. Then, we introduce some novel animal models of osteoporosis and take a perspective view of the requisite needs for large animal models of osteoporosis.