31 January 2007, Volume 27 Issue 1

    

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  Foundation, Heredity and Function Test of Double Gene Mouse with hMCP and hCD₅₉

  WEI Qmg-xin1,2,CHEN Shi3, WEI Yan1,2, ZHEN Xin-min1,2, LI Li1,2, QIAO Xian-feng1,2,LIU Xi-mei1,2,ZHOU Jing-rong1,2, TIAN Yong-xiang1,2

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 1-5.

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  Objective Comparing protective effect on the cell from one CRP or two CRPs. Method Double gene mouse was founded by mixed injection. Mixed human MCP and CD₅₉ gene was injected into pronuclear of 521 mice zygote. Results In all of the newborn mouse ,there are 14 with CD₅₉ gene, 12 with MCP gene, 7 with double gene. Total transgene rate was 5.0 percent. Five double gene founders mated with nontransgene mouse, which their baby had 14 with MCP gene, 16 with CD₅₉ gene, 10 with double gene corresponded to 33.3 percent of baby mouse. The hearts in vitro were taken out to be perfused with fresh human blood plasma, the average heart beating time was 138 ± 25 min in the hCD₅₉/hMCP group. The time is significantly longer than that of other group(78 土 27 min in the K3
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  Establishment and Evaluation on Modified Athesclerotic plaque Model in Rabbit

  HUANG Cheng-lei1,LI Tian-qi1,QIAO Wei-wei2,LUO Xin-ping1,LI Jian1,NI Huan-chun1,SHEN Wei1,LI Hong-zhi3,JIN Bo1,SHI Hai-ming1

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 6-10.

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  Objective To establish the modified athesclerotic plaque model and evaluate it compared with traditional animal model. Methods New Zealand rabbits were fed with high cholesterol diet and feromal artery balloon injury were conducted to establish the improved atherosclerotic plaque model, PT and endothelium thickness, foam cell area were determined, and TIMI was monitored. Result Compared with the traditional group, modified plaque model group presents significant stenosis of the femoral artey，shortening of the PT and thickening of the endothelium. Conclusion The modified plaque model, which is accurately controlled and easily repeated, could be used to evaluate the progress of atherosclerosis and demonstrate the mechanism of the plaque.
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  Establishment of Sleep Apnea Syndrome Model in Rats Induced by Intermittent Hypoxia Exposure

  DU Xiao-yan1;HU Liang-gang2;FAN Xiao-fang2;CAI Xiao-hong3;ZHOU Yong-hai3;JlA Shan-shan2;GONG Yong-sheng2

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 11-14.

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  Objective To establish a sleep apnea syndrome (SAS) model in rats. Methods Fifty male SPF Sprague-Dawlev rats were randomly divided into five groups: normal control group (NC)， 2-week and 4-week experimental control groups (2C and 4C)，and 2-week and 4-week intermittent hypoxia groups (2H and 4H). The rats of 2H and 4H groups were laid in the hypoxic cabin for 8h per day，in which nitrogen and oxygen were input circularly (90s/cycle). The Fi02max was 21%± 0.5% and Fi02min 9% ± 1.5%. The gas concentration and the cyclic time were automatically controlled. The rats of 2C and 4C groups were in the normoxic cabin, NC group in room air. At the experimental end points，the right ventricle flmctions (RVEDP, RVESP，RV ± dp/df max), the mean pulmonary arterial pressure (mPAP), the mean carotid arterial pressure(mCAP)，the weight ratio of left ventricle plus septum to body [(LV+S)/WT] and of right ventricle to left ventricle plus septum [RV/(LV+S)] were measured, respectively. Results The mPAPs of 211 group and 4H group were much higher than those of 2C group and 4C group? respectively (P＜ 0.05). The RV+dp/dt, RV-dp/dt and RVESP in 4H group were significantly higher than those of 4C and 2H groups(P＜ 0.01). The mCAP，RV/(LV+S) and (LV+S)/WT among all of the groups were no significantly different (P＞ 0.05). Conclusion The sleep apnea syndrome SPF rat model can be successfully established with the method by which operation is convenient, control is precise，repeatability is well and the effects is well consistent with the SAS pathophysiology.
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  Study on Phamacodynamic Properties of L-VCR and Its Anti-tumor Effect

  WANG Xiao-chen1;WANG Xiang-wei1;QIN Yan1;XU Zhi-ru1;WANG Guang-feng1;LAI Qing-qin1;YANG Zhen-mao2;LIU Quan-hai1

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 15-19.

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  Objectives To analyze the pharmacodynamic properties of liposome Vincristine (L-VCR), its pharmacological effects against tumor and toxicology. The characteristics of L-VCR and the possible mechanism of its actions were discussed. Methods Wistar rats were inoculated with Walker256 tumor. Blood drug level was analyzed by HPLC and pharmacodynamic parameters were calculated using computer software. Mice were inoculated with B16 melanoma and L-VCR was given i.v. Tumor inhibi-toiy rate and non-tumor body weight was calculated. The LD50 for L-VCR was determined in both rats and mice. Results Compared with Free Vincristine (F-VCR), the plasma and tumor level of L-VCR was higher and sustained significantly longer. Its AUCQ25-t was increased about 121 times after treatment. L-VCR at 2 — 0.5 mg/kg dosage could significantly reduce the weight of the tumor mass in all three cases mentioned above and certain does-effect correlation could be observed. Compared with that of F-VCR, the LD50 of L-VCR was significantly higher in animals. Conclusion L-VCR could significantly extend the half-life of VCR and increase the AUC. Its effects on transplantable tumors was significantly stronger compared to that of F-VCR while its toxicity was significantly lower. The effective period of VCR was prolonged due to the coating of liposome and its level in tumor was increased. The drug was released slower and its toxicity was therefore reduced.
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  Isolation, Culture and Characterization of Primary Rat Muscle Satellite Cell

  LI Ying-chuan;DING Qiang;FANG Zu-jun

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 20-24.

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  Objective To discuss isolation and culture methods of primary rat skeletal muscle satellite cells and observe its characterization of proliferation and differentiation in vitro culture. Methods Primary muscle satellite cells of relative high purity were obtained by improved two-steps enzymatic digestion combined with pre-plating techniques. Marker of muscle-derived cell, desmin, was identified by histocytochemistry. The characterization of cell proliferation was studied by the way of MTT assay experiment. Differentiation of muscle satellite cell under different culture conditions in vitro was also studied. Results The purity of harvested muscle satellite cell was more than 90 percent. When cultured in vitro, the latent phase of the cell is at the lst to 2nd days, platform phase is at the 5th to 6th day. It shows strong proliferative ability in vitro culture and could fuse with each other into myotube cells without special induction. Conclusions Muscle satellite cells by in vitro culture were enough to meet study of tissue engineering and cell transplantation.
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  Peripheral α1 and α1 Receptors Involved in Stressful Hypertension -Induced Inhibition of Carotid Sinus Baroreceptor Reflex in Rats

  WANG Guo-qing;SHEN Xin-e;WANG Lin-hui;ZOU Rong;ZHOU Xi-ping

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 25-28.

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  Objective To investigate the effects of peripheral oti and Ol2 receptors on the stressful hypertension-induced carotid sinus baroreceptor reflex (CSR) resetting. Methods Twenty-three male Sprague-Dawley rats were subjected to unavoidable electric foot-shock twice daily for a week, each session of foot-shock lasted 2 hours. The left and right carotid sinus regions were isolated from the systemic circulation under anesthesia with pentobarbital sodium. The intracarotid sinus pressure (ISP) was altered in a stepwise manner in vivo. ISP- mean arterial pressure (MAP) relationship curve and its characteristic parameters were constructed by fitting to the logistic function with five parameters. We observed the changes in CSR performance resulted from stressful hypertension and the effects of injection with Ol1 or Ol2 receptor selective antagonist, phenoxybenzamine (PBZ) or yohimbine (YOH), into the peripheral vein on the responses of CSR to stressful hypertension. Results Peripheral veinal injection (iv.) of PBZ (2.5 jumol/L, in 0.15 jul/g) significantly shifted the ISP-MAP relationship curve downwards (P＜/i>＜0.05) and obviously decreased the value of the reflex parameters such as threshold pressure, saturation pressure and ISP at maximum gain (P＜0.05), but increased maximum gain (P＜0.05) in the stressed rats. The effects of iv. YOH (5 jumol/L, in 0.15 ul/g) on the changes in CSR induced by stressful hypertension were similar to those of PBZ, but the effect of YOH was less remarkable than that of PBZ (P＜0.05). Peripheral injection with the corresponding dose of PBZ or YOH in the unstressed rats did not change CSR performance significantly (P＜/i>＞0.05). However, injection of PBZ or YOH could not completely abolish the stressful hypertension-induced changes in CSR performance. Conclusion Peripheral oti and (Ol2 receptors are involved in the stressful hypertension-induced CSR inhibitory resetting. Peripheral Ol1 receptor may play more important roles in this resetting and at the same time, the effects of stressful hypertension on CSR also have other mechanism.
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  The Effect of rhBMP-2 on the Expression of MMP13 and Collagen-2 in mRNA Level during Osteogenic Differentiation

  XU Lan;FAN Yan;ZHOU Ying-hui;WU Shi-liang

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 29-31.

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  Objective To investigate the interaction of metalloproteinase-13 (MMP-13) and type II collagen (Col-2) in C3H10T1/2 cells, and explore the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on the expression of MMP13 and Col-2 in mRNA level during osteogenic differentiation induced by rhBMP-2. Methods rhBMP was utilized to induce osteoblastic differentiation of C3H10T1/2 cells. Total RNA was isolated at various time points to detect the expression of MMP13 and Col-2 by the method of quantitative Real-time PCR. Results There was a negative correlation between MMP-13 and Col-2 expression in C3H1 OTI/2 cells without rhBMP-2 treatment in the first 10 days. In the rhBMP-2 treated cells, however, expression of MMIP-13 was inhibited and there was a negative correlation between MMP13 and Col-2 expression in the first 14 days. Conclusions There is a obvious correlation between the expression of MMP13 and Col-2. rhBMP-2 inhibits the expression of MMP13 while it induces osteogenic differentiation of C3H10T1/2 cells.
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  Experimental Study on Bladder Tumor Model in Wistar Rats

  DING Hong1;QIAN Li-xin2

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 32-36.

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  Objective To study the dynamic processes of bladder tumor in Wistar rat induced by intravesical irrigation of the carcinogen,MNU and discuss how to establish an appropriate animal model of bladder tumor. Methods Sixty Wistar rats were randomly divided into 3 groups. The model rats were intravesically affused with MNU as carcinogen to induce bladder tumor every two weeks. Rats were sacrificed in different periods. Results In total of 60 rats , 58 of them had finished the experiment while 2 rats died during the period.In cancer induced group(VINU group) there were 39 survived during the experiment in which 37(92.5%) of them were confirmed by pathology to be bladder cancer and other 2 rats(5%) were atypical dysplasia(hyperplasia).Thirty three rats were diagnosed to be positive bladder tumor in 37 rats combined with urinary HE and AO pigmentation while the positive ratio is 89.2%. Conclusion Bladder tumor induced by MNU is highly resemble with that of human beings in both histologic and pathologic character.lt is expected to have good foreground to diagnose bladder tumor in animal model combined with urinary HE and AO pigmentation.
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  Establishment of a Modified Orthotopic Renal Transplantation Model in Rat

  ZHAO Yan-zong1;DU Wei-cheng1;GUO Xi-tao1;YUE Zhong-Jin2

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 37-40.

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  Objective To establish a modified model of orthotopic renal transplantation in rat. Methods SD rats was used as donors and Wistar rats as recipients, respectively. The left orthotopic transplantation was performed. The donor’s mesenteric superior artery and recipient’s renal artery was anastomosed by a modified sleeve technique. End-to-end anastomosis of recipient's renal vein to the donor’s inferior vena cava via an temporary intraluminal tube. Urinary tract reconstruction was accomplished by suturing the donor ureter with a bladder patch to the recipient bladder. Results In 61 rats, the total time of operation and arteiy anastomotic time was (161±11) minutes and (11±2) minutes, respectively. The success rate of operation was 95.1 percent and survival time of recipients were 4 to 17 days. The surgical complications including anastomosis hemorrhage, artery thrombosis, urine leak and acute rejection were observed. Conclusion This is a simple and practical technique for establishment of renal transplantation model and can be applied in the experiment of renal transplantation research.
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  Preliminary Research in Establishment of Esophageal Carcinoma Heterotopic Transplantation model in SCID Mice

  XIE Yang-min1,2,XIE Liang-xi3,LI De-rui3,LI fang2,CHEN Jiong-yu3,HONG Chao-qun3,ZENG De-nian1

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 41-44.

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  Objective To report a new method for the establishing spontaneously metastatic animal model of human esophagus carcinoma and the metastatic-lymph node-deriving subline. Methods SCID mice were used in the experiments. Human esophagus carcinoma cell line Eca-109 were injected into the gastric mucosa layers in mice with a concentration of (1-2) X 10⁶ in 0.2 ml solution. All mice were sacrifed 3 months later or when the mice were found under the condition of general failure. Tumor metastases were investigated pathologically，part of the metastastic lymph node were inoculated subcu-taneously for the establishment of metastatic sub line. Results We established 9 gastric implantation animal models, one of which were found to have lung and lymph node metastases. Part of the metastatic lymph node was implanted sub cutaneous ly in mice, and after its survival in vivo for 4 passages, a new subline was established using primary tissue culture. Conclusion Gastric wall implantation is a feasible method for establishing spontaneous metastases animal model and metastatic sub line of human esophagus carcinoma.
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  Induction of Transplantation Tolerance by CTLA4-Ig Adenovirus Gene Local Transfection in Allogeneic Rats

  PING Ji-gen,YAN Chun-yin,LU Jin-xing

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 45-47.

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  Objective To induce renal transplantation tolerance by CTLA4-Ig adenovirus gene therapy in allogeneic rats and study the relative mechanisms of transplantation tolerance.Methods The kidney of SD rats were transfected with CTLA4-Ig gene-recombined adenovirus.After the orthotopic renal transplantation between SD and Wistar rats was conducted,all the rats were divided into 3 groups:group I was the control;rats in group II were perfused with empty EGFP and there in group Ⅲ with CTLA4-Ig adenovirus. The allograft survival time and the allograft funtion between the three groups were compared. Result The survival time was significantly longer in rat of group Ⅲ than in other 2 groups(31.3 ±6.8 days vs 8.2 士 1.2 days and 8.0 土 1.6 days). The serum creatinine of transfection group at 7th and 14th postoperative days was lower than that of controls at 7th postoperative day. Conclusion Local transfection of CTLA4-Ig gene can augment the tolerance and prolong the survival time of renal allograft significantly.
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  Review on Lentivirus Transgenesis for Producing Transgenic Animal

  HUANG Li-zhen1,LIU Guang-ze2,GU Wei-wang1

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 63-66.

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  Lentiviruses belong to the retroviruse and they are characterized by infecting non-dividing cells except for sharing many characteristic features with simple retroviruses. The recent studies found that the transgenic efficiency was enhanced significantly with lentiviral transgenesis. Due to its high efficiency，this method will make a breakthrough in transgenic animal research field. This article is about the principle of constructing lentiviral vector, the common approach of producing transgenic animal with lentiviral trans genes is and its latest progress home and abroad.
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  Discussion on Genetic Control of Captive Nonhuman Primate Colonies

  SUN Xiao-mei,YE You-song,TONG Pin-fen,DAI Jie-jie

  Laboratory Animal and Comparative Medicine. 2007, 27(1): 67-70.

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  Genetic control is an important component of the general management of nonhuman primate colonies. In this paper, some approaches were reviewed for the goals of genetic control including pedigrees, pedigree validation, quantitative traits and estimating genetic variability. Genetic control is critical for maintaining variability and avoidance of inbreeding, ensuring the long-term viability of captive nonhuman primate colonies.