Table of Content

31 March 2007, Volume 27 Issue 3

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Establishment of a Standard Acute Myocardial Infarction Model for Cellular Transplantation Research in Rats

WANG Jin-yi1,FAN Hui-min1,LIU Zhong-min1,FU Xue-lian2,Wang Jun-chen2,ZHAO Shu-guang1,KONG Xiang1,LU Wei1

2007, 27(3):  149-154. 

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Objective To explore the feasibility of establishment of acute myocardial infarction (AMI) models induced by occlusion of left anterior descending coronary artery (LAD), which provide a reliable model for the study of repair myocardial infarction by cellular transplantation in rat. Methods Fourteen adult Sprague-Dawley (SD) rats with respirator were established by ligating the middle-proximal 1/3segment of left anterior descending coronary artery (LAD) in AMI model group. Continuous electrocardiographic monitoring were performed both before the LAD ligating and after the LAD ligated by multiple peripheral adapter (MPA) multichannel physiologic recorder system. The cardiac function indicated by these parameters of left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and 士 dp/dtmax were observed by multipurpose polygraph of langendorff perfusion of the isolated heart methods, and their pathological morphology, myocardial infarction size (MIS) and mortality were detected and analyzed four weeks after LAD occlusion to see whether there were any tissue necrosis and its extension. Other ten adult SD rats without ligating coronary artery as control group. Results The survival rate of the treated rats was 71.43% (10/14) beyond four weeks. After LAD ligation, electrocardiogram displayed ST elevated continuously immediately and pathologic Q wave after ligating 30 min. Pathologic examination suggested myofibers chaotically arranged, myofilament ruptured or lysed, nuclei pyknosis and fragmentation, interstitial hyperemia and edema, infiltration of inflammatory cells, necro-myocardium were superseded by fibrous tissue. Meanwhile, the cardiac function observation indicated by these parameters of LVSP、LVEDP and 土dp/dtmax descend markedly in LAD occlusion rats after ligating four weeks late. MIS were invariableness in AMI model group (44.38% 士 8.04。/。）. Conclusion A standard rat AMI model could be created by surgical ligation of LAD, and the model would be suitable for the research on cellular transplantation.

Effect of μ-Opioid Receptor Antisense Oligodeoxynucleotide on Methamphetamine-Induced Behavioral Sensitization in Mice

SHEN Xin-e1,WANG Guo-qing1,ZHOU Xi-ping1,ZOU Rong1,MA Tan-geng2

2007, 27(3):  155-159. 

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Objective To investigate the effect of u-opioid receptor (MOR) on the process of methamphetamine (METH) - induced behavioral sensitization in mice. Methods The antisense and mismatch oligodeoxynucleotides(ODN) for MOR gene were synthesized and delivered individually to mice by intracerebroventricular injection (i.c.v.). Locomotor activity was detected after acute administration with METH in mice. Effects of repeated intermittent administrations of METH on locomotor activity were measured to observe the effects of MOR on the development of behavioral sensitization to METH. Results MOR ASODN could attenuate the METH-induced hyperlocomotor activity in mice and MOR ASODN-exposed mice failed to sensitize to the METH challenge. Conclusion MOR may play an important neuromodulatory role in METH- induced behavior sensitization.

Effects of Tangxinkang on NF-κB of Diabetic Cardiomyopathy in Rat

YAO Li,SUN Yang,BAI Yu-bin,LIU Chun-hong,LIU Cheng-gang,WANG Bing,LIU Zheng,ZHOU Xue-ming,PAUL Featherstone,JIANG De-you

2007, 27(3):  160-162. 

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Objective To evaluate the therapeutic effects of Tangxinkang on diabetic cardiomyopathy in rat and discuss its mechanism .Methods The model rats were established by ip STZ and hyper-calorie diet. The glucose metabolism、insulin、myocardium histomorphology and nuclear factor-kappa B (NF-kB) were studied and the therapeutic mechanism in biology of Tangxinkang treating the diabetic myocardial damage were also discussed. Results (1) Tangxinkang has good effects on decreasing glucose and HbAlc in blood and increasing the level of insulin. (2)Tangxinkang has good repairing effect on damaged cardiac muscle and pancrease in histomorphologic change.(3)Tangxinkang has obvious adjustment role on over-expressed NF-kB of diabetic cardiac muscle (P＜0.01). Conclusion Tangxinkang has an obvious therapeutic role on diabetic cardiomyopathy through adjusting the over-expressed NF-kB of diabetic cardiac muscle mainly.

Study on the Relationship Between Memory CD8+T Cell and GVHD

QIU Ling1,2,HE Shan2,CAO Qi2,ZHANG Yan-yun2,HUANG Rui1

2007, 27(3):  163-167. 

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Objective To study the relationship between memory CD8⁺ T cell and GVHD by using a mouse model of CD8^??+ T cell-mediated acute graft versus host disease(GVHD). Methods C3H. SW(H-2D^b, CD45.2^?+) mice were used as donors and B6/SJL(H-2D^b, CD45.1⁺) mice as recipients, both of them are MHC-identical but miHA-mismatched. Before transplantation, the B6 recipients received ^{137y ray}(9.5 Gy) TBI administered in three times from a ¹³⁷Cs source, then the T cell-depleted bone marrow (T" BM) cells and CD8⁺Tcells from C3H.SW donors were injected immediately via tail vein. The weights of recipient mice and dead rate were calculated after transplantation. Results The quantitative changes in the livers and spleens of recipient mice of the donor CD8⁺T cells(CD45.2^?+), donor CD8⁺T cells that produced IFN-Y and the apoptosis related molecular annexin V on donor CD8⁺T cells were analyzed by Flow Cytometry. In the miHA-mismatched CD8+ T cell-mediated acute GVHD mouse model, the recipient mice began to die 28 days following transplantation and the dead rate was 58% until 70 days. The analysis showed that in parallel to GVHD development, donor CD8⁺T cells and CD8⁺T cells secreting high levels of IFN-Y peaked in the livers and spleens of recipient mice by day 14，considerably declined by day 28, increased again by day 42 after transplantation. The decline in donor CD8⁺T cells was accompanied with increased annexin V-positive donor CD8⁺T cells. These results suggest that a typical memory T cell response，as manifested by T cell expansion, contraction, and re-expansion, develops in recipients during the GVHD process，and a small population of reactive CD8⁺T cells survives after the death phase of effect cells. Conclusions The MHC-identical but miHA mismatched mouse model is analogous to human clinical miHA-mismatched bone marrow transplantation. There is the relation of the donor CD8⁺ T cells to GVHD persistence.

Pathological Observation on Beagle Dogs Infected with Ehrlichia

LIU Shi-zhong1,2,Gu Wei-wang1,TANG Bo-heng2,Na Shun-ba-ya-er3,MA Yu-hai2,Zen Zhao-zhi1,WU Qing-hong

2007, 27(3):  168-171. 

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Objective To study pathological changes on Beagle dogs infected with Ehrlichia. Methods six beagle dogs that had been artificially infected with Ehrlichia were examined By using routine pathological methods. Results The obvious lesions were of subcutaneous hemorrhage and stagnation of blood in the major tissues and organs, and severe swollen enlargement in spleens and livers. The histopathological changes were characterized by accumulations of lymphocytes and plasma cells in blood vessels in livers, spleens, lungs and kidneys, and accumulations of lymphocytes and plasma cells in perivascular of livers. Conclusion The observation on pathological changes coincide mostly with reports abroad, but the macroscopic changes is more severe. This may contribute to the difference in pathogenicity of the strain in China.

Establishment on Infant Rats Model of Irritable Bowel Syndrome and for its Primary Exploration

ZHU Xue-ping,CHEN Wei-chang,XU Zheng,WU Xu-dong,ZHU Shu-yang,ZHU Xiao-li

2007, 27(3):  172-175. 

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Objective To establish an infant rat model of irritable bowel syndrome with “integrative methods”，and test its visceral sensitivity by two different ways. Methods Sixteen SD neonatal rats were divided into model and control group randomly. The model was established by separated from mother rat，binded up mildly and intrarectal administration of mustard oil and so on; the control group didn’t intervene. After the intervention, let the rats have a rest for 2 weeks. The threshold of abdominal withdrawal reflex (AWR) was evaluated during rectal distension by self-made dilator in different tensions, and the changes of rectal sensitivity were identified by the abdominal electrical activity measured at postnatal 30 days. At last pathological changes of the rectal mucosa was measured. At the beginning of intervention, the two groups’ feces were scored according to the Bristol pattern, collected the feces and measured their wet and dried weights. Results The model group showed a significant decrease (P＜0.01) in the volume threshold of abdominal withdral reflex during rectal distension; there was a significant increase of abdominal electrical activity responding to different distension pressure compared with control rats (P＜0.01). The colon’s pathological changes had no significant difference in the two groups. The feces of model group were mostly soft mass or muddy, Bristol Scores was 5.3 土 0.8; which of control group were delicately sausage or mass, Bristol Scores was 3.6 土 0.5，the difference was significant in the two groups. The wet and dried weights of the model ’s feces were 10.8 ±1.2、 3.2 士 0.4 g respectively, and the control’s were 5.4 土 0.3、3.6 土 0.7 g respectively, there was significant difference in the two groups，wet weights(P＜0.05). Conclusions The “integrative” methods (concluded to separation from mother、mechanism fetter and chronic rectal irritation with mustard oil) to the infant rats may establish the children’s irritable bowel syndrome model，of which mechanism is visceral hypersensitivity possibly.

Establisment of B7-DC Transgenic Mouse

LIU Ying1,YUE Ting2,SHI Yun-song3,YUAN Wen-yue3

2007, 27(3):  176-179. 

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Objective Establishment of B7-DC transgenic mouse model. Method B7-DC cDNA was cloned by RT-PCR from mouse thymus, then it was inserted into PEF6/V5-His vector and get PEF6-B7-DC/V5-His which contain a fuse protein B7-DC-V5-His. The PEF6-B7-DC/V5-His gene was microinjected to the pro nuclear of FVB mouse embryos, then transfer the embryos to the foster mother and get new individuals. Positive founders was confirmed by PCR and Southern. Result Eighty three injected embryos were transferred and 39 offspring mice were obtained, three of them are positive and can be successfully inherited to F2. Conclusion The B7-DC transgenic mice model was established.

Preparation of Tilapia Hepatocytes by Cold Digestion of Collagenase

ZHAO Zhong-xin,LI Tie-yan,LIU Yan-fang,YANG Chao-wu,ZHENG Xing-li,WEI Zheng,HUANG Wei

2007, 27(3):  180-182. 

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Objective To investigate the method of isolation and extraction of tilapia hepatocytes. Methods The fish liver was washing repeatedly by D-Hank, then the liver was cut into small pieces. The tissues was filtered and centrifuged after digested by Collagenase IV. Then the suspensions were purified by Percoll isolation medium after washed by DMEM. Hapatocytes was counted. The morphologic characteristics of the isolated hapatocytes were accessed and observed by light microscope. Result An amount mixed cells were found through the cold digestion of Collagenase method. Further isolation was done by centrifuged at 90 g, 60 g and 30g. Conclusion The method of Cold digestion by Collagenase had obtained the best isolated result. Better purification and amount of hepatocytes could be done by centrifugation with 60g for 2 minutes and centrifugation with 30g for 3 minutes.

Surveillance and Analysis of Pasteurella pneumotropica Infection in Mice

GAO Zheng-qin1,XING Jin1,WANG Chun-ling1,ZHANG Qiang2,HE Zheng-ming1

2007, 27(3):  183-185. 

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Objective To investigate and analysis Pasteurella pneumotropica infection in laboratory mice. Methods Two hundred and one specimens from the air, wind port, door knob, drinking water, feeds，bedding，feces, and pharyngolarynx of surveillance mice (NIH, KM, ICR and C57BL / 6 mice from a animal facility) were tested by bacterial cultivation, colony morphology, Gram-staining, biochemical test，and the slide agglutination test. Results Among 201 tested specimens, 5 mice pharyngolarynx swab, 1 drinking water and 1 animal feeds were identified contaminating with P. pneumotropica. Conclusion The healthy mice in the facility were infected with Pasteurella pneumotropica. And the drinking water and feeds were also contaminated with Pasteurella pneumotropica. Therefore, the survey should be further strengthened for prevention and control pathogenic microorganism in laboratory animals.

Effects of Four Nutrition Metabolism Related Genes on Blood Biochemistry Index of Mouse

AO Hong1,XU Lan-wen1,YANG Ping1,JIN Mei-lei2

2007, 27(3):  186-189. 

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Objective To investigate the effects of four nutrition metabolism related genes(DL、 Tr、Pe、P3) on blood biochemistry index of BALB/c mouse. Methods Antisensenucleic acids of the four genes with microamount quantitate were injected into the lateral cerebral ventricle of the mouse. The experiment was divided into three groups: antisensenucleic acids group, random sequence nucleic acid control group and saline control group. At the fourth day after thrice injection, using the blood serum to determine the blood biochemistry index. Results Comparing with control group, six indexes of DL and Tr group has significant difference respectively, four and three indexes of Pe and P3 group has significant difference respectively. The triglyceride in blood of the three antisensenucleic acids group (DL、Tr、and P3) BALB/c mouse was significantly higher than the control group, especially the cholesterin of Tr antisensenucleic acids group was obviously higher than the control group, and all were beyond the range of normal values. Moreover, the glucose in the blood of the three antisensenucleic acids group (DL、Tr、and P3) was significantly higher than the control group and beyond the range of normal values. Conclusion The Tr genes have a relation with lipid metabolism, and the DL、Tr、 and P3 genes have also a relation with glycometabolism.

Effects of Guanxinning Tablet on Myocardial Oxygen Consumption in Dogs

PAN Yong-ming,YING Hua-zhong,CHEN Min-li,XU Jian-qin,ZHOU Wei-min,WANG Hui,XU Xiao-ping,Wang De-jun,L(U) Jian-min

2007, 27(3):  190-191. 

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Objective To study the effects of Guanxinning tablet on myocardial oxygen consumption in dogs. Methods Beagle dogs were divided randomly into control, Guanxinning tablet high dosage (GXNTH), media dosage (GXNTM), low dosage (GXNTL) and Guanxinning injection groups. Myocardial oxygen consumption was determined. Result Compared to the control group ,the myocardial oxygen consumption of GXNTH and GXNTM were reduced significantly (P＜0.05，P＜0.01). Conclusion Guanxinning tablet can reduce the myocardial oxygen consumption and ameliorate the supply-demand relationship of myocardial oxygen.

Develpment of Animal Models for Prostatitis

2007, 27(3):  211-214. 

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There are mainly two kinds model of prostatitis: bacterial and nonbacterial prostatitis. E.coli as the primary etiologic agent causes acute and chronic bacterial prostatitis. Nonbacterial prostatitis is often induced by chemical agents or biological angents. Compared with chemic-induced model, the biologies-induced prostatitis has more specific, which is an ideal prostatitis model. The concrete methods include estradiol-indued and male accessory gland(MAG) extracts in complete Freund’s adjuvant (CFA)-induced prostatitis. These models can provide methods and evidences for basic and clinical research.