30 June 2009, Volume 29 Issue 6

    

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  Establishment of Rabbit Renal Interstitial Fibrosis Model By Unilateral Ureter Ligation

  MA Dan1’2，PAN Xmg-hua1，YU Yue-mmg1，HONG Yu1, YANG Ren-hua1，MALi-hua1, DING Ya-nan1, ZHU Xiang-qmg1，CHEN Qiang1

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 357-360.

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  Objective To establish the model of rabbit renal interstitial fibrosis. Method The renal interstitial fibrosis model was established in rabbit by unilateral ureter obstruction. Serum creatinine (Scr) and urea nitrogen (Bun) level of the rabbits from 3 groups were analyzed at the end of the first, second and third week and the SPECT scanning were adopted to monitor the change of renal function; at the end of the fourth week the pathological change of renal tissue were observed by HE and PAS. Result The content of serum Scr and Bun are both significantly increased, comparing with those of the control group rabbit (P＜0.05). SPECT showed the ill side kidney with significant deformation, renal imaging faint and glomerular filtration rate significantly lower than those of control group. The interstitial fibre hyperplasia, fibrosis changes, chronic inflammation cells infiltration, part of renal tubular epithelial abscisin, cloudy swelling, part of renal tubular atrophy，and glomerular blooding were observed in experimental group, but no abnormal in control group. Conclusion The model rabbit induced by unilateral ureter ligation showing obvious renal interstitial fibrosis character and may be fit for the clinical research.
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  Proliferation Effect of HGF and EGF on Hepatic Stem Cell Cultured from Rat in vitro

  YANG Li-pmg1, MA Jun-xun2, YUAN Ju-fang1, SHAO Jun-shi1, WANG Hong-bao

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 361-365.

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  Objective To research the effect of time and dosage of offering HGF and EGF comprised of different concentration, which can establish foundation of fundamental and clinical research for liver disease. Methods Hepatic stem cells isolated from portal vein perfusion were cultivated by serum-free medium to observe the effect of HGF and EGF of different concentration and time on proliferation of hepatic stem cell in rat. Results Hepatic stem cells from adult rat have small volume and its outline form and nucleus were round or oval-shape. Proliferation effect in 5—160 ng/ml EGF group and 10-160 ng/ml HGF group was significantly stronger, compared with control (P＜0.05); and when HGF was to 20 ng/ml, its effect was the most obvious (P＜0.01); In all HGF added with EGF(10 ng/ml) groups, except for 1、5 ng/ml group，proliferation effect were greater than control (P＜0.01). In time proliferation effect of hepatic cell treated with HGF (20 ng/ml), with the time prolongation, there exsited manifest effect and was up to the peak at the fifth day; at the seventh day, there happened little decrease. For the time proliferation effect with EGF (10 ng/ml), there emerged the peak at the seventh day. There was no difference at the fifth and seventh day. Proliferation effect of HSC combinated with 20 ng/ml HGF and 10 ng/ml EGF showed that there was not obvious change at the first day; and was up to the peak at the third day. Conclusion There was manifest improvement for the hepatic stem cells cultured by serum-free of SD mouse treated with EGF and HGF. There existed synergistic effect between two factors for the hepatic stem cell proliferation in vitro.
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  Effect of Fusion Conditions on Efficiency of Electrofusion of Mouse Somatic Cell Nuclear Transfer

  WANG Yi1’2，JU Hui-mmg2，XUE Zheng-feng1’2，Yang Lmg1-2,FENG Ya-jie2，GUO Lei2, YUAN Yu-guo2，CHENG Yong2

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 366-371.

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  Objective To improve the efficiency of electrofusion of reconstructed embryos derived from oocytes and fetal fibroblast cells both of B₆D₂F₁strain within a mouse somatic cell nuclear transfer（SCNT）program.Methods The effect of different pre-treatment on donor cells，fusion midia，electrical field intensity，electrodes’shape and cell passage on the efficiency of electro fusion was studied.Results（l）The pre-treatment of PEG/DMSO on donor cells significantly improved the fusion rate（51.11%vs 25.64%）（P＜0.05）；（2）Fusion rate in the fusion media containing O.lg/L polyvinyl alcohol（PVA）was higher than those in the media without PVA.Fusion rate in fusion media containing 0.32 mol/L mannitol（48.21%）was higher than those in the media with 0.28 mol/L，0.30 mol/L and 0.36 mol/L mannitol（27.78%，41.67%and 30.11%）；（3）Although the chamber is a much more routine way for fusing the reconstructed embryos，the tip shape electrodes can improve the fusion rate（42.31%，43.13%）comparing to routine parallel line electrodes（30.43%，33.33%）；（4）the efficiency of electrofusion of oocytes with fetal fibroblast cells at the found passage and the first passage was higher than that at the second and the third passage（38.98%，38.46%，36.73%and 35.00%），regardless of other fusion conditions.Conclusion Both the pre-treatment of PEG/DMSO on donor cells and the addition of PVA to the fusion media can improve the efficiency of electro fusion in the procedure of mouse SCNT.
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  Objective To investigate the Mamu-A*Ol allele of Chinese rhesus monkey in our monkey colony. Methods The blood sample (200 ul) were collected from femoral vein of 101 monkeys borm in 2007 randomly selected from the colony and the genomic DNA was extracted. The primers were designed according to the specific sequence of Mamu-A*Ol. The allele was investigation by sequence specific primers polymerase chain reaction. Results Six monkey were selected that carried Mamu-A+01 allele among the 110 monkeys, but the homology was only 96% with the Indian rhesus monkey，s corresponding sequnce. Conclusions The Mamu-A*Ol allele exist in Chinese rhesus monkeys, but was different with the Indian rhesus monkey, further work should be done to elucidate the function of the Mamu-A*01 allele.

  SUN Zhao-zeng, LI Ai-xue，HU Zhong-ming, SHI Yan-sheng, BAI Jie-ying, ZENG Lin

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 372-374.

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  Objective To investigate the Mamu-A*Ol allele of Chinese rhesus monkey in our monkey colony. Methods The blood sample (200 ul) were collected from femoral vein of 101 monkeys borm in 2007 randomly selected from the colony and the genomic DNA was extracted. The primers were designed according to the specific sequence of Mamu-A*Ol. The allele was investigation by sequence specific primers polymerase chain reaction. Results Six monkey were selected that carried Mamu-A+01 allele among the 110 monkeys, but the homology was only 96% with the Indian rhesus monkey，s corresponding sequnce. Conclusions The Mamu-A*Ol allele exist in Chinese rhesus monkeys, but was different with the Indian rhesus monkey, further work should be done to elucidate the function of the Mamu-A*01 allele.
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  Determination of Haematological, Blood Biochemical Parameters and Organs Weight in Cricetulus Migratorius

  WANG Hong1, LIAO Li-fu2，XU Yi-mei2，LIU Shuang-Huan1，Ma L卜 Ymg1，YUE Bmg-Fe 11，XING Riu-Chang1，HE Zheng-Mmg1

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 375-378.

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  Objective To determine the normal ranges of blood physiological and biochemical data, and to determine the body weight and the weight of organs in Cricetulus Migratorius and compare for variations of these data between the sex. Method The blood samples collected were tested for physiological values by using MEK-7222K Hematology Analyzer, biochemical values by using OLYMPUS AU400 auto-analyser. The body weight and the weight organs were determined and the organ coefficient were calculated. Result The sexual difference of 8 weeks Cricetulus Migratorius was noted in physiological values such as WBC、RBC、HGB、HCT and RDW(P＜0.05) other than MCV、MCH、 MCHC、PLT、PCT、MPV and PDW (p>0.05)，and also noted in biochemical values such as AST、 BUN、GLU and CREA(P＜0.05) other than ALT、ALB、ALP、TP、CA、P、CHO and TG (p>0.05). Sexual difference of 8 weeks Cricetulus Migratorius was noted in organ coefficient such as spleen、right kidney、right adrenal glands、lungs and heart (P＜0.05)，and also noted in organs weight such as spleen (P＜0.05). Conclusion The normal physiological, biochemical data and the organ coefficient in Cricetulus Migratorius are affected by a variety of factors. Some of the data are significantly different between the both genders.
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  Comparasion on Blood Biochemical Indices among Three Breeds of Rabbit

  YING Hua-zhong, ZHU Ke-yan，CHEN Mm-li，SHOU Qi-yang, LV Jian-mm, PAN Yong-ming,

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 407-410.

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  Objective To comparative analysis of blood biochemical indexes in WHBE rabbits, Japanese White rabbits and New Zealand ^White rabbits，and further reveal on blood biochemical characteristics of WHBE rabbits. Methods Fourty in each of WHBE rabbits, Japanese White rabbits and New Zealand White rabbits，half male and female，the blood was sampled from the central artery of ear for testing the blood biochemical indexes. Results (1) Gender factor has significant influences on serum Ca, P, CHOL，HDL-C，LDL-C，FMN indexes m WHBE rabbits (P＜0.05,尸<0.01); (2) Compared with Japanese White rabbits and New Zealand rabbits, partly blood biochemical indices had significant differences between WHBE rabbit and Japanese White rabbits or between WHBE rabbit and New Zealand White rabbits，those maybe related with itself characteristics;(3)CHOL, HDL-C, LDL-C, UA, CK and CK-MB were obviously exist with differences between human being and WHBE rabbit, and the other biochemical indexes were similar to those of human being. Conclusion The WHBE rabbits may be used for research on partly change of biochemical indices and metabolism diseases in human.
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  Measurement of Reticulocytes and Its Subpopulation in WHBE Rabbits

  ZHOU Wei-mm, PAN Yong-mmg, CHEN Mm-li，WANG Hui, LIN Lm, WANG De-jun

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 411-414.

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  Objective To determine and comparative analysis of reticulocytes and its subpopulation in WHBE rabbits，Japanese White rabbits and New Zealand White rabbits. Methods Fourty in each of WHBE rabbits, Japanese White rabbits and New Zealand White rabbits, half male and female, were used. The anticoagulant blood samples from the central artery of ear, and then reticulocytes parameters indexes were determined by Bayer ADVIA 120 automatic blood analyzer. Results (1) Gender factor has significant influences on RDWr，HDWr，CHDWr，RDWm, CHDWm, RDWg, CHDWg indexes m WHBE rabbits (P＜0.05，P＜0.01); (2) Reticulocytes parameters in WHBE rabbits had significant differences between Japanese White rabbits and New Zealand White rabbits, for example, Retie % was lower in WHBE rabbits than that of Japanese White rabbits and New Zealand White rabbits(P＜0.01); (3)RPI in WHBE rabbits was similar to that of human being, compared with human reference. Conclusion WHBE rabbit has its own reticulocytes and its subpopulation characteristics, and may be used as model animal for research on anemia disease etc. in human.
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  Analysis and Determination of Hemorheology and Blood Coagulation Function in WHBE Rabbits

  SHOU Qi-yang, CHEN Mm-li, ZHU Ke-yan, PAN Yong-mmg, PEN Dmg-guo, WANG Hui, LIN Lin, LV Jian-min

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 415-418.

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  Objective To determine and comparative analysis of hemorheological parameters and blood coagulation function in WHBE rabbits，Japanese White rabbits and New Zealand White rabbits，and further to reveal on hemorrheological and blood coagualtion system 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 samples from the central artery of ear，and then hemorrheological parameters and blood coagulation function were determined by automatic hemorheology analyzer and CA-500 automatic blood coagulation analyzer，respectively.Results（1）Gender factor has significant influences on hematocrit，the erythrocyte sedimentation rate，K value of erythrocyte sedimentation rate equation and erythrocyte accounts in WHBE rabbits（〈i>P＜0.05，P＜0.01），but has no significant influence to blood coagulation of WHBE rabbits（尸>0.05）；（2）Compared with WHBE rabbits，the hemorheological parameters of WHBE rabbits have no significant difference to Japanese White rabbits（尸>0.05），but PT，PT/R，PTINR of female WHBE rabbits were obvious higher than those of female Japanese White rabbits（P＜0.05），and PT%was lower than that of female Japanese White rabbits（P＜0.05）；while whole blood viscosity at low shear rate，Fbg and TT of male WHBE rabbits were higher than those of male New Zealand White rabbits（P＜0.05），hematocrit was obviously lower in WHBE rabbit than those of New Zealand White rabbits（P＜0.05），moreover，the whole blood viscosity，capillary plasma viscosity，blood reduced viscosity at low and high shear rate，erythrocyte deformation index，erythrocyte electrophoresis index，casson viscosity and PT%were higher in female WHBE rabbits than those of female new Zealand White rabbits（P＜0.05，P＜0.01），while hermatocrit，K value of erythrocyte sedimentation rate equation，PT，PT/R and PTINR was significant lower than those of female New Zealand White rabbits（P
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  Determination and Analysis of Electrocardiogram in Anesthetized WHBE Rabbits

  CHEN Liang, PAN Yong-mmg, CHEN Mm-li，ZHANG Li-zong, XU Jian-qm, YING Hua-zhong

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 419-423.

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  Objective To investigate whether the WHBE rabbit was suitable for experimental electrocardiographic observation. Methods The adult 12 WHBE rabbits, 7 Japanese white rabbits and 6 Chinchilla rabbits were anesthetized by sodium amylobarbitone (30 mg/kg) and the electrocardiograms (ECG) of the rabbits were recorded by multi-channel physiological recorder. Results The basic wave pattern of the EGG of WHBE rabbit was similar to that of human being. Heart rhythm of WHBE rabbit was sinus original. Owing to the fast heart rate of WHBE rabbit (288.89士28.14 beats per minute, bpm), the P-wave of the WHBE rabbit ECG usually occurred on the descending branch of T-wave. The Q-T interval and K-value of the WHBE rabbits were both lower than those of Japanese white rabbits and Chinchilla rabbits. The range of the ECG axis in WHBE rabbits (40.00±36.93 degrees) were changed greatly. Conclusion The shapes of waves in chest leads of WHBE rabbit are obvious and regular. The recorded voltage was higher. So the ECG in chest leads is more available for the clinical analysis, and WHBE rabbits would be more suitable as a new kind of laboratory animals for electrocardiographic experimental studies.
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  The Generation and Application of Transgenic RNAi Mice

  LIN Ju-fang1, SHI Hui-juan2, ZHAO Jian1

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 424-428.

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  RNA interference (RNAi)has been extensively used for sequence-specific silencing of target gene. The generation of RNAi transgenic mice made it possible to knock down gene expression in mammalian species. This paper reviewed the generation of transgenic RNAi mice and its application. The contribution of RNAi transgenic mice to functional genomics and of its prospect for application were also discussed.
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  Animal Model of Mycobacterium Tuberculosis and the Application

  LIU Fang1，YANG Hua1, ZHOU Wen-jiang1-2, FAN Xiao-yong1

  Laboratory Animal and Comparative Medicine. 2009, 29(6): 429-429.

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  With the publication of the report on WHO 2009 Global Tuberculosis(TB) Control, deep study on TB is extremely uigent. Traditional research couldn’t satisfy the needs of TB’s situation today, such as the problem of MDR/XDR-TB which has became into focus of the people. Ideal animal models of MTB take important meaning for studying the immunopathogenesis of TB, as well as evaluating the effects of novel anti-TB drugs, vaccines and diagnostic reagent. According to different purpose of the study and application，here just given an overview of various animal models of Mycobacterium tuberculosis (MTB) which have been built, and also take a view of their future..