Table of Content

31 January 2010, Volume 30 Issue 1

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Application of GFP/Luc Double-tagging Technique in Tumour Model 0f Mouse

WANG Xiao-min1, YAN Ming-xia1, LIANG Lin-hui1, LI Jin-zhi2, ZHU Miao-xin1, WU Yi-qian1, YAO Ming1

2010, 30(1):  2-7. 

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Objective To establish human lung cancer cell line and human liver cancer cell line with GFP/Luc double-tagging which having high expressing rate，and to observe development of the tumors in deep organisms of the nude mice with in vivo biofluorescence imaging. Methods The SPC-A-1-GFP/Luc cell line and SMMC-7721 -GFP/Luc cell line were established by lentivirus transfection, en-hancing their GFP/Luc double-tagging expressing rate to about 100% by flow cytometer, then were inoculated in lung and liver of the BALB/c nude mice respectively, finally, the behaviors of the two cell lines transfected stably in development of the tumours was detected by in vivo biofluorescence imaging.Results The cell lines of SPC-A-1-GFP/Luc and SMMC-7721-GFP/Luc with GFP/Luc double-taking have been built up with the expressing rate nearly 100%. After the two cell lines have been inoculated in lung and liver of the nude mice, the tiny tumor focuses in early time and the status of tumors in late time in the deep organisms were observed by the in vivo biofluorescence imaging. Conclusion The estab-lished cell lines with stable GFP/Luc double-tagging, coupled with the application of m vivo biofluorescence imaging, can detect the tumors in deep organisms of the nude mouse intravitaly，real-timely, quantitively,and may be offering an ideal device for researchers to further explore development of the tumor.

Establishment of Infection-type Periventricular Leukomalacia Models in Neonatal Rats

HE Ya-fang, CHEN Hui-jin, QIAN Long-hua, CHEN Guan-yi

2010, 30(1):  8-11. 

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Objective To establish the animal model of infection-type periventricular leukomalacia (PVL) in two-day-old SD rats. Methods Forty-eight 2-day-old SD rats were randomly divided into 2 groups consisting of the sham group and the PVL group. Under stereo-positioner of brain, the lipopolysac-charide (LPS ) 1 ug/g was injected into the lateral ventricle of the neonatal rats in the PVL group, and similar volume of sterile PBS was injected into the lateral ventricle of neonatal rats in the sham group. The pathological assessment under light microscope and the immunohistochemical analysis of 04-positive preoligodendrocytes (preOLs) were respectively undertaken in all neonatal rats 5 days after LPS injection.Results The obvious injuries of white matter and the massive loss of 04-positive PreOLs induced by LPS were observed by the pathological assessment under light microscope and the immunohistochemi-cal analysis. Conclusion The infection-type PVL model of neonatal rats which corresponds to the pathological features of PVL in human premature infants can be successfully established by the intraven-tricular injection of LPS.

Establishment of Orthotopic Colon Tumor and Metastasis Model by Coloclysis of MNU in Rat

TANG Mao-sheng, HU Zhi-qian，WANG Wei-jun, YAO Hou-shan, PENG huan, WANG Qiang

2010, 30(1):  12-16. 

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Objective To establish an orthotopic colon tumor and metastasis model by the coloclysis of MNU in SD Rat, to observe the carcinogenesis development and the metastasis process. Methods SD Rats were given the MNU clyster，the clinical symptom was checked, the tumor growth, metastasis of the rat were observed. To respectively sacrifice 12 SD rat every time at 8, 12, 16 weeks later, observe the tumor in situ and the metastasis. Evaluate the effect of this colon cancer model through pathological methods. Result None of the SD rat died, 8 weeks later，clinical symptom show hematochezia and lymphadenectasis，16 weeks later, 91.7% rat formed colon neoplasma and many lymphadenectasis，pathological examination confirm the colon adencarcinoma with lymph node metastasis. Conclusion The successful establishment of orthotopic colon tumor model by the coloclysis of MNU in SD Rat. The induction process is convenience and utility, the model has the colon carcinoma autogenesis development successfully.The orthotopic tumor and metastasis model provide useful tools for the study of mecha-nism of metastasis and its treatment of colon cancer.

Observation on Micro- and Ultra-structures of the Liver in Zebrafish(Brachydanio rerio)

CHU Xiao-hong1, PU Jun-yi1,2,3, BAO Hui-jun1, HU Jian-hua2,GAO Cheng2, YAO Yi-lin1，XU Chun-sheng1，QIN Jun-hui1, CHENG Qiu-sheng1

2010, 30(1):  17-23. 

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Objective To observing and analyze the microstructures and ultrastructures of the liver in zebrafish. Methods The tissue samples were observed by using light and electron microscopy.Results There was no typical portal area within the parenchyma, bile ducts and venules were randomly distributed within the parenchyma. The polygonal hcpatocytes are arranged in twisting, branching and anastomosing cords usually two cells thick .There was a single big and spherical nucleus in the hepatocyte,and the nucleoli displayed a separation into granulose component, fibrillar centers and dense fibrillar component. The cytoplasm are dominated by numerous stacks of non-fenestrated lamellae of rough endoplasmic reticulum (RER) of considerable length arranged in close parallel array and smooth endo-plasmic reticulum was rarely observed. The number of mitochondria is of considerable, and myelinbody was found in mitochondria. When the cytoplasm was rich in mitochondria, RER, and ribosome and the number of nucleopore was massive, glycogen was found in small concentrations, and a few lysosome may be observed. When glycogen was found in large concentrations, the number of nucleopore was few, so the cytoplasmic organoids were. These suggested that the consumption of glycogen ensured normal metabolism of nucleic acids and proteins, and mitochondria provided energy in the process of protein synthesis. The biliary system is arranged in arborization. Bile canaliculus includeded intracellular and intercellular bile canaliculus，the intracellular bile canaliculus had lots of long microvilli, the intercel-lular bile canaliculus had relatively shorter ones. Erythrocytes, lymphocytes，fat-storing cells and endot-helial cells were also described. A comparison between zebrafish liver and that of other teleosts was made. Conclusions The way of hepatocytes arranged in zebrafish was different from other teleostei and was not exactly conformable to previous accounts, this distinction was relevant to the animal species and research methods. The ultrastructure of hepatocytes proved its active synthesis and metabolic function.

Isolation and Identification of Helicobacter pylori from Guizhou Miniature Pigs

GAO Zheng - qin1，ZHANG Qiang2, YUE Bing-fei1，HE Zheng-ming1

2010, 30(1):  24-28. 

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Objective To isolate and identify Helicobacter pylori in Guizhou miniature pig .Methods Specimens of an abnormal growth and development retarded miniature pig was collected.Bacteria isolation and culture，biochemical identification, susceptibility test，serum agglutination test，PCR assay were conducted. Results One strain of Helicobacter pylori (HPp0812) was isolated and identified. The genome of HPp0812 strain could be amplified by 16S rRNA gene of Helicobacter genus and ure A gene of Helicobacter pylori specific primers in PCR assay. Through determination of the nucleotide sequence, the highest sequence homology of this strain of bacteria could be demonstrated with Helicobacter pylori reference strain in GenB epidemiological ank and it was found to be 99%.Conclusion The strain HPp0812 appears to be the Helicobacter pylori first isolated from miniature pig in China, and it provides reliable scientific data for further research and investigation.

Cryopreservation of Embryos From HBV Transgenic and Cataract Mice in a Vitrification Solution

KONG De-qiang1, SUN Zhao-zeng1, HU Zhong-ming1, ZHU Shi-en2,ZHAO Yan-bin1，ZHANG Hui-yang1, SHI Yan-sheng1，HUAXiu-chun1，ZENG Lin1

2010, 30(1):  29-32. 

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Objective To study the value of open pulled straw(OPS) method on cryopreservation of embryos between HBV transgenic mice and cataract mice (CM). Methods Ethylene glygol (EG) is used as the cryoprotectant, OPS method is adopted to carry out vitrification freezing for 3.5d embryo of CM，P₂₁^HBxPHBx and KM mice, and compared the numbers of embryo，recovery rate and developmental rate after defrosting of KM, CM and P₂₁^HBxPHBx.Results The average embryo numbers of KM , CM，and P₂₁^HBxPHBx 31.12,19.32，9.14 respectively . The recovery rates are 88.7%, 81.1% and 80.3% respectively.The developmental rates are 82.6%，79.4% and 56.6% respectively. The embryo numbers ofP₂₁^HBxPHBx are lower than those of KM . The recovery rates are not significantly different in three kinds of mice .The embryo developmental rate after defrosting of P₂₁^HBxPHBx is lower than that of KM. Conclusion The superovulated numbers and developmental rate after defrosting 0f P₂₁^HBxPHBx are iower than those of KM，and there is no significant difference between CM and KM.

Study on Biological Characteristics of Spontaneous Type 2 Diabetic db/db Mice

LI Na, ZHANG Zhou, FENG Ying

2010, 30(1):  33-38. 

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Objective To compare the biological characteristics of spontaneous type 2 diabetic C57BL/KsJ-db/db mice in growth curve, glucose metabolism, serum insulin, weight of major organs，and organ coefficient with C57BL/Ks J-db/+ mice as the control, then the morphologic changes of the liver, kidney and pancreas were observed. Methods Forth weeks old mice were used with total of twenty weeks experimental period, the weight and glucose at each week, serum insulin at every two weeks were measured. After the experimental period, the mice were sacrified, and the organ and fat were weighted. The pathological changes of the liver, kidney and pancreas were observed. Results ① The weight, glucose and serum insulin of db/db mice were higher than those of the control group at the same age(PP<0.01). But the spleen，pancreas and kidney was less relatively, especially of the spleen and pancreas in female mice(P<0.01).③ The liver appeared significant cell putrescence, adipose degeneration, inflammatory cell infiltration and fibroplastic proliferation in db/db mice. The renal interstitial appeared hyperaemia, dropsy, hyaline cast in renal tubules, and adipose degeneration of epithelial cell. At the same time，the number of glomeruli pancreatici and islet cells decreased obviously. Conclusions The db/db mice is a suitable disease model of type 2 diabetes mellitus, which shows hyperglycemia, high insulin resistance，insufficiency of islet function, renal and hepatic alterations.

Morphological Observation on Main Anatomical Structures of Canine Hearts Cavity

WEI Ke-ke1，SHEN Biao2，ZHANG Hai-tao1，ZHANG Sen-lin1，YE Bin1,ZHU Hua-nan1, ZHANG Jiao3, CUI Xiao-zhi3，CHENG Xiao4

2010, 30(1):  39-43. 

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Objective To observe and measure the main anatomical structure of canine hearts cavity,and to accumulate data for the study of comparative zootomy. Methods The hearts from fourty-five dogs fixed with 10% formaldehyde for one week were dissected, then were observed and measuredrelative datas of the mitral complex tricuspid complex tricuspid valve ring aortic orifice pulmonary veins in front of and behind vena cava coronary sinus and fossa oval is dealed with statistics then analyzed the datas and compared with human heart. Results 1. The left ventricle of canine heart was a cone. It was divided into the gateway of left ventriclc and outlet of left ventricle. The main structure of the gateway was mitral complex, which was composed of the mitral ring and valve chordae tendinease and papillary muscles. The length of mitral ring was 49.16 ± 10.00mm. The average number of chordae tendineass was 16.72 ± 2.79. The main structures of the outlet of left ventricle were aortic valve aortic sinus and aortic orifice. The length of aortic orifice was 34.55 ± 8.00 mm. 2. The main anatomical structures of the right ventricle in canine were made up of the tricuspid complex pulmonary valve false tendon and septomarginal trabecula，et al. The tricuspid complex was built up of the tricuspid valve ring and valve chordae tendinease and papillary muscles. The average length of tricuspid valve ring was 71.03 ± 14.20mm. The average length of pulmonary valve ring was 35. 97 ± 8.17mm. 3. The main anatomi-cal structures of the right and left atrium in canine were composed of the before and after vena cava pulmonary veins coronary sinus and fossa ovalis. Before and after the inferior vena cava the perioral diameter were 43.91 ± 10.17mm and 37.67 ± 10.54 mm. The average of pulmonary veins was 29.44± 12.71mnr Coronary sinus circumference was 14.12 ± 4.00 mm. Coronary sinus which was round the mouth accounted for 89.7%, the lamiaceae accounted for 10.3%. Fossa ovalis was found in 100%.Conclusion The main anatomical structure of the Canine hearts cavity is similar to that of human heart in spite of the difference in size and the total number of left ventricular false tendon and chordae tendinease.

Determination and Comparison of Hematological Indices in WHBE Rabbits

YING Hua-zhong, SHOU Qi-yang, CHEN Min-li, PAN Yong-ming,WANG Hui, LIN Lin, ZHU Ke-yan, WANG De-jun, XU Xiao-ping

2010, 30(1):  44-47. 

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Objective To determine and comparative analysis of hematological parameters in WHBE rabbits, Japanese White rabbits and New Zealand White rabbits，and further reveal on hematological characteristics of WHBE rabbits. Methods Fourty in each of WHBE rabbits, Japanese White rabbits and New Zealand White rabbits，half male and female, the anticoagulant blood was sampled from the central artery of ear, and then hematological parameters indexes were determined by Bayer AD VIA 120 automatic blood analyzer. Results (1) Gender factor has significant influences on neutrophil, lymphocyte, MCH, MCHC and RDW m WHBE rabbits (P<0.05, P<0.01);(2) Leukocytes amounts was lower in WHBE rabbits than that of Japanese White rabbits (P<0.01),lymphocyte and hemoglobin were higher in WHBE rabbits than those of Japanese White rabbits and New Zealand White rabbits (P<0.01).Conclusion WHBE rabbit has its own special characteristics, also has its population characteristics in common, and the hematological parameters in WHBE rabbits were similar to those of human being.

Rodent Models of Colorectal Tumor and Evaluation System of Drugs

ZHOU Wei-Mrn1，SHENG Hong-Qiang2, CHEN Wen-Bin3, CAI Yue-Qin1，CHEN Min-Li1

2010, 30(1):  60-67. 

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In the present paper, common rodent models for colorectal cancer are systematically discussed and reviewed with regard to preparation methods, experimental observation means and drug evaluation system, etc. Additionally, the few disadvantage of animal model of human colorectal cancer are analyzed. The review will be beneficial for research in fields of colorectal cancer concerning carcino-genesis mechanism, chemopre vent ion, drug development.

Progress in Research on Outer Capsid Protein and Receptor of Mammalian Reovirus

SONG Yu1，2,3，4,ZHOU Jie2,3,4, GAO Cheng2,3,4，HU Jianhua1，2，3,4

2010, 30(1):  68-72. 

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It is a key step that Mammalian reovirus adhesion and entry into the host cells during the infection，including that the different domains on the outer capsid protein interact with the different receptors on the surface of host cells. There are three kinds of virus capsid protein ol，o3,and ul，playing important roles of the binding of reovirus to receptors oil the surface of host cells. The reovirus strains use Sialic Acid(SA), junctional adhesion Molecule-1, cell surface carbohydrate as the receptors，which could bind to the surface protein of the virus specifically and explain both cell-type specific tropism and broad hosts, thus, the research on the virus-receptor interactions are conducive to reveal the pathogenesis and replication. This review will summarize recent advances made in studies of reovirus outer capsid protein and Receptor.

Advances on Developmental Toxicity of Cadmium in Zebrafish Embryos

HU Xiao-qing1,2，LI Wei-hua2，JIN Tai-yi1，DING Xun-cheng2

2010, 30(1):  73-76. 

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Cadmium is a widespread non-essential heavy metal in occupational and living environment.It is toxic to multiple organs in animals. This review is given on advanced data on potential developmental toxicity as well as the mechanisms of cadmium in zebrafish embryos which provides a reference for further studies on toxicological effects of cadmium in the development of other vertebrates.