Abstract:Objective To establish a hygromyein-resistant transgenie BDF1 model for preparation of feeder layers of hygromyein-resistant ES cells clones. Methods Hygremyein B phosphotransferase gene fragments were mleroinjected into the zygote male pronuelei of BDF1 mice. Altogether 169 zygotes were injected and 129 zygote cells were transplanted into the oviduct of aceeptor pregnant female mice. Results Genome DNA from mice tail were detected by PCR and Southern blot showing 9 of 37 were positives transgenie mice. One of the founders was analysed by RT-PCR and found that hyg was expressed in the kidney, muscle and spleen. Conclusion A hygromyein-resistant transgenie BDF1 mouse model has been successfully established. This model will provide favorable fundamental conditions for gene knock-out studies.

Abstract:目的制备乳腺特异性高表达人促红细胞生成素（hEPO）转基因奶山羊。方法采用牛β-乳球蛋白基因（BLG）调控元件和hEPO全长编码序列基因组DNA构建真核表达载体，应用受精卵原核注射的方法制备hEPO转基因山羊。结果在原核注射获得的188头羔羊中，经Southern blot法检测有4头羊含有hEPO基因，其中3头为母羊，1头公羊于出生后20d死亡；3头转基因母羊hEPO基因的拷贝数分别为1、10、2；Western blot检测结果显示转基因羊乳中的hEPO分子质量为32kDa；MTT法检测结果表明，在泌乳10d的3只转基因羊乳汁中，每毫升乳汁中hEPO活性分别达到1.17×10^2IU、1.90×10^4IU、1.91×10^4IU。结论牛BLG能够调控hEPO基因在山羊乳腺中高表达，为实现其他药用蛋白在山羊乳腺中表达奠定了基础。

Abstract:Objective To study the mechanism of polymorphism of Hbb gene at single nucleotide polymorphism(SNP)level. Methods Hbb gene was analyzed in 3 aspects,i.e. DNA,RNA and protein polypeptides. Results For the genome DNA,four SNP loci,two T(G) of exon 1,G(A) of exon 2 and A(G) of exon 3,were detected correlating with Hbb d/s haplotype. For the RNA,four SNP loci,corresponding to two T(G) of exon 1,G(A) of exon 2,and A(G) of exon 3,were found correlating with Hbb d/s haplotype. For the protein polypeptide,three interchanges of Cys/Gly,Ser/Ala,Thr/Ala at position 13,20 and 139 were detected correlating with Hbbd and Hbbs haplotypes,which corresponded to two T(G) of exon 1 and A(G) of exon 3,respectively. Conclusion Three interchanges of Cys/Gly,Ser/Ala,Thr/Ala at position 13,20 and 139 might provide the basis for the two haplotypes,Hbbd and Hbbs.

Abstract:目的将兔出血症病毒（RHDV）VP60全长基因在昆虫细胞-杆状病毒系统中表达，验证重组蛋白形成病毒样颗粒（VLPs）的能力及其生物学特性，探讨VLPs作为检测抗原及亚单位疫苗的潜力。方法用Bac-to-Bac系统体外表达RHDVVP60全长基因。以免疫荧光及Western blotting检测蛋白表达情况及确定蛋白最佳表达条件；免疫电镜观察VLPs形态，并对VLPs的血凝性、免疫原性进行检测。结果SDS-PAGE电泳分析表明，表达的重组蛋白分子量大小约为68KDa，在免疫荧光、琼脂扩散、ELISA试验中均与RHD多克隆抗血清特异性反应；接种重组病毒的Sf9细胞裂解液在电镜下可观察到与RHDV形态相似的VLPs；该VLPs可凝集人“O”、“B”型红细胞，凝集可被RHD多克隆抗血清所抑制；含VLPs的Sf9细胞裂解液可不经纯化用作间接ELISA抗原，所建立的ELISA方法与进口商品化试剂盒相比，特异性良好，敏感性、检出率稍低；将含VLPs的细胞裂解液加氟氏佐剂免疫兔，HI效价可达1∶40，可经受致死量病毒攻击。结论RHDV-VLPs的获得及其良好的免疫原性，为RHD血清学检测试剂的标准化、亚单位疫苗研制应用奠定基础，同时在转移载体及RHDV受体方面研究亦有潜在应用价值。

Abstract:目的分离犬MC2R基因cDNA5′末端，分析其启动区域特点。方法采用了RNA连接酶介导的RACE（RLM-RACE）技术分离了犬MC2R基因和局部序列比对工具（Basic Local Alignment Search Tool，BLAST）对CDS区进行了初步验证。结果新分离了犬MC2RcDNA的5′末端，并对其启动区序列作了初步分析。序列分析显示，该基因至少由两个外显子（exon1和exon2）组成，exon1和exon2的一部分编码5′非翻译区（5′-UTR），exon2其余的部分编码整个编码区。结论克隆了犬MC2R基因的5′末端，在其启动区发现了inr、SF-1、SP1、CRE、PPRE、AP-1等多个顺式作用元件，为犬MC2R表达调控研究奠定基础。

Abstract:目的建立绿色荧光蛋白转基因小鼠模型，并采取胚胎冷冻的方法进行保种。方法通过原核显微注射法，把线性化、纯化后的外源基因pEGFP注射入BDF1小鼠受精卵中，胚胎移植给同期发情的假孕受体母鼠，获得子代小鼠。经鉴定对有表达的转基因鼠进行胚胎冷冻保种。结果移植注射胚胎385枚给30只假孕小鼠共出生了306只后代鼠，经PCR和southern blot检测得到5只阳性小鼠。F2代转基因鼠胚胎冷冻240枚胚胎。结论通过显微注射法使外源基因pEGFP在小鼠基因组中得到整合，建立了转pEGFP的转基因小鼠模型。

Abstract:目的建立KLK1转基因大鼠模型。方法腹腔注射PMSG-HCG（150-150IU/kg）超排不同周龄（4-8周）SD大鼠比较超排效果的差异，以可用于注射胚胎数作为评判标准确定最佳超排周龄。构建pBC-klk1转基因构件，经酶切、纯化后通过显微注射方法导入SD大鼠受精卵原核并移植到同期受孕的SD受体母鼠输卵管内。出生后仔鼠用PCR和Southern方法检测鼠尾DNA鉴定基因型，通过RT-PCR和免疫组化方法检测klk1基因表达。结果4-8周SD大鼠超排后分别获得受精卵14±14.8、30±15.2、13.3±13.7、13±14.7、8±5.7，5周龄大鼠超排效果最好，与其他周龄大鼠相比有显差异，P〈0.05；显微注射1538枚卵，移植685（44.53%）枚卵于31只受体输卵管中，12（12/31，38.7%）只怀孕，共产仔62只，经PCR检测获得6只阳性鼠，Southern检测3只阳性。对Southern检测阳性转基因大鼠子代进行RT-PCR检测和免疫组化分析证明klk1基因在肾脏、胰腺和乳腺内表达。结论成功建立klk1基因表达的转基因大鼠模型，该模型是高血压病研究的理想动物模型。

Abstract:目的建立615小鼠标准染色体组型与G带染色体核型，提供可靠的细胞遗传学背景资料。方法成年615小鼠8只，雌雄各半，提取骨髓细胞，制片，镜检。确立615小鼠体细胞染色体数目。选择10个典型细胞测量染色体基本数据。G带染色。结果615小鼠的染色体数目为40条，XX为雌性，XY为雄性。所有染色体均为中部着丝点。X染色体相对长度仅次于第1对染色体，Y染色体的相对长度在第4对染色体和第5对染色体之间。G显带条数与小鼠有很大差异，接近于大鼠。结论615小鼠的核型为2n=40=2×19m＋（x）m＋（y）m，G显带共262条。

Abstract:目的测定SX1近交系小鼠的生长繁育指标，为建立该品系小鼠的生物学特性资料提供实验数据。方法从采用全同胞兄妹交配方式繁殖获得的F20、F21代仔鼠中选择30对小鼠，连续繁殖5胎，分别测定窝产仔数、24h窝重、仔鼠均重、离乳仔数、离乳均重、胎间隔等几项指标；以第2胎小鼠作为生长发育观察对象，测定1-10周间每周的体重变化，计算每周体重增长情况。结果SX1小鼠1-5胎平均窝产仔数5.42±0.76只；平均窝重10.36±1.45g；仔鼠均重1.93±0.07g；离乳仔数5.21±0.95只；离乳窝重61.41±9.67g；离乳均重11.98±0.50g；离乳成活率96.09%；平均胎间隔33.63±0.68。1-10周平均每周增重雄鼠3.08±2.40g，雌鼠2.76±2.01g。3-5周时为生长发育旺盛期，平均每周增重雄鼠5.95±1.41g，雌鼠5.18±0.96g。结论SX1近交系小鼠具有稳定的繁殖性能，幼鼠离乳成活率高，各项主要指标基本符合近交系小鼠的特征。

Abstract:目的探讨2型糖尿病证病结合动物模型的造模方法及成模标准，建立2型糖尿病病证结合动物模型。方法应用STZ造成大鼠糖尿病（FBG≥16.7mmol/L）后应用中药造成中医阴阳两虚；阴虚热盛；气阴两虚；血瘀气滞证候模型。结果（1）造模后出现饮水增多、尿量增多、生长迟缓、身体消瘦、拉尾排便、排尿等糖尿病大鼠共同特征。阴阳两虚组大鼠还出现大便干燥、舌红等征象；阴虚热盛组大鼠出现大便干燥、舌红等征象；气阴两虚组大鼠出现精神萎糜、倦怠懒动、舌胖大，少津；血瘀气滞组大鼠还出现背毛减少、臀部毛色枯黄、性情暴烈、易激惹、捕捉时叫声频繁、抵抗力大、攻击行为频繁、大便质稀色深、舌质暗等血瘀气滞气滞的症状。（2）实验室指标有不同程度改变。结论应用传统的糖尿病造模方法与糖尿病病证特点和中药四气五味药性理论相结合，研制2型糖尿病病证结合动物模型具有可行性。

Abstract:Objective To investigate the methods of controlling microorganism contamination and keeping clear in laboratory animal rooms of barrier system. Method To improve the air clearance methods of turbulent air flow in laboratory animal rooms of barrier system by setting non-dynamic facilities which can change the turbulent air flow modality in animal room into unidirectional way. Results In turbulent air rooms the average of bacterial sedimentation was 47, the floating bacteria was 154.5. In unidirectional air rooms the bacterial sedimentation was 1.5 and the floating bacteria was 7. Conclusion The unidirectional air flow is better than turbulent air flow. It will provide technical data for formulating new environmental standards of laboratory animal barrier system in future in our country.

Abstract:目的通过对二种血瘀证大鼠模型的内皮素（ET）、一氧化氮（NO）和血液流变学指标进行比较，探讨其在血瘀证量化诊断中的意义。方法SD大鼠50只，分别建立气虚血瘀证和气滞寒凝血瘀证动物模型，测定二种模型血浆ET和NO水平，用LBY-N6B全自动自清洗血流变仪对二种模型进行血液流变学的分析。结果（1）与模型对照组比较，气虚血瘀模型组全血黏度低切变率、毛细管血浆黏度均明显增加（分别为P〈0.05），血浆ET水平显著升高（P〈0.01），NO水平显著降低（P〈0.01）；（2）与空白对照组比较，气滞寒凝血瘀模型组全血黏度低切、中切、高切以及毛细管血浆黏度均明显增加（分别为P〈0.01），红细胞聚集指数增加（P〈0.05），血浆ET水平显著升高（P〈0.01），NO水平显著降低（P〈0.01）。结论内皮细胞损伤，ET和NO比例失衡可能是血瘀证量化诊断的重要指标。

Abstract:Objective To discuss the mechanism of action of bDNF on the intelligence of aged mice. Methods 90 Kunming mice over 55 weeks old were selected and randomly divided into groups. To observe the expression of choline acetyltransterase (ChAT-IR) and in the labyrinth experiment of each group before and after administration with integral experimental technique and immunohistochemistry. Results Significant changes of ChAT-IR AchE and labyrinthian experiment occurred (P<0.05) in the cytidine diphosphocholine group. Exogenous bDNF treatment group changed more significantly compared to CHAT AchE and labyrinth experiment (P<0.01). Conclusion bDNF can improve the intelligence in aged mice.

Abstract:Objective To study the application of cryopreservation of embryos of transgenic FVB mice expressing antibacterial peptides. Methods Superovulated FVB female mice and living for 4 - 6 weeks, 6 - 8 weeks, 8 - 10 weeks, respectively, were mated with heterozygote male mice. The 2-cell amphicytulas were collected and cryopreservted. One week later, the cryopreservted 2-cell amphicytulas were thawed and transplanted. Result From the 140 cryopreservted mouse embryos,98 viable embryos were obtained after thawing. Conclusion Embryo cryopreservation and embryo transfer technology can be used for transgenic mouse preservation.

Abstract:Objective The genetic variations of FUT1 gene at nucleotide position 307 of its open reading frame were investigated by PCR-RFLP in a total of 245 pigs from 3 exotic pig breeds and 3 Chinese native pig breeds. The results showed that the genetic polymorphisms of the FUTI locus were only detected in the 3 exotic pig breeds. There were 3 different genotypes and the majority were the susceptible forms including GG and AG. All Chinese native pig breeds only presented the susceptible genotype GG.

Abstract:The screened environment of laboratory animal institution is an isolated system at dynamic running. When people, animal, feedstuff, air, padding and articles enter into this area, strict microorganism control should be carried out. If the equipment control element of transferring compartmeni, aqueduct, anemon-showering cave and buffer cave are lack of necessary door forbiden auto control equipment, it may causes fault work and lapses facility in daily work. This door forbidden system can control the time and the switch program of equipments, and start disinfection.

Abstract:Objective To study the effect of Opuntia on the blood glucose level in rat model of type II diabetes mellitus. Method Rat model of type II diabetes mellitus (DM) was eastablished with streptozotocin (STZ). 60 rats with DM were divided into six groups (10 rats each): normal control (N), DM control (A), PEDG (B), and low dose (E, fed with 3 g/d), moderate dose (D, fed with 6 g/d) and high dose (C, fed with 9 g/d) of Opuntia. Results The blood glucose levels in C (9.21 0.85), D (11.35 0.91) and E(14.40 1.30) groups were significantly decreased in comparison with that before Opuntia administration in groups C and D (P<0.01), and also significant difference compared with group A (P<0.01), but not with group B (P>0.05). Conclusion Opuntia may decrease the blood glucose level in rat model of type II diabetes mellitus, and is worth further studing on its clinical application.

Abstract:Objective To observe the effects of pentobarbital sodium,chloraldurat and ethylcarbamate on hematology in male SD rats. Method Male Sprague-Dawley (SD) rats were anesthetized by intraperitoneal injection of pentobarbital sodium, chloraldurat and ethylcarbamate, respectively. Serum biochemical parameters and blood cell indexes were determined within 20 minutes after injection. Results Some blood biochemical parameters and blood cell indexes of anesthetized rats showed significant difference in comparison with normal rats. Conclusion Anaesthesia can affect the hematology considerably. Therefore, scientific researchers should select anaesthetics properly.

Abstract:Cryobiology is a jumped-up subject which investigates the influence of low temperature on organism. This extensively studied and applied technique not only promotes the progress in biology, medicine and other sciences, but also will create huge benefits for agriculture, stockbreeding, pharmacy and food manufacturing. In particular, the cryobiology has engendered far-reaching influence on environment protection, to severe endangered animal salvation and biodiversity conservation. Laboratory animals are important experimental materials for life sciences. Several scientific trends have increased the large number of mouse models and placed an intolerable load on the financial and infrastructural resources needed to maintain these models as conventional breeding colonies in recent years. The cryobiology has been applied for breeding, preservation of genetic resources, and development of new strains of laboratory animals. Anyway, the preservation of mice model will establish the furthersome condition for further studies of mammalian genetics and difficult and complicated diseases.

Abstract:Objective In order to evaluate the safety of use of per acetic acid(PAA)and sodium dichloroisocyanurate (ACL-60)toxicity. Method To Ccompare the results of acute and subacute toxicity test. Result The results of acute oral LD50 showed that PAA and ACL-60 are of low toxicity grade. PAA and ACL-60 disinfectants could cause moderate irritation to animal skin and eye, and 2 g/L peracetic acid was non-irritative to animal skin and slightly irritative to the eye. 112 mg/L ACL-60 could cause moderate irritation to Guinea pig skin. The rates of PCE micronuclear cells induced by 3 dosage were not significantly different from that of negative control group (P>0.05). Conclusion peracetic acid and sodium dichloroisocyanurate are mildly toxic and can be considered as safe disinfectant commonly used at low concentration.

Abstract:Objective To observe the effect of rabbit sacculus rotundus peptides on virulent Newcastle disease virus (NDV). Methods Virulent NDV was treated with rabbit sacculus rotundus peptides, and inoculated into chicken embryo. The erythrocyte agglutination titer of virulent NDV in chicken embryo allantoic fluids was measured, and pathological changes of chicken embryo were examined. Results The erythrocyte agglutination titer of virulent NDV was lower than that of control series, and the pathological changes were less serious than those in control series, indicating that rabbit sacculus rotundus peptides could inhibit the pathological changes of chicken embryo caused by virulent NDV. Conclusion The rabbit sacculus rotundus peptides has inhibitory effect on virulent NDV in vitro.

Abstract:Objective To establish and compare the reference range of normal values of five types of lymphocyte subsets in mice of three strains by flow cytometry. Method The five types of Th1/Th2 cytokines in Balb/C, C3H/HeJ and C3H/SW mice were determined by flow cytometry. The results were statistically analyzed with SPSS 10.0 software. Resoults The distribution of the five lymphocyte subsets presents a Gaussian distribution in the 3 mouse strains, respectively. The 95% confidence interval for mean values can be accepted. The difference at lymphocyte subsets level is not significant in Balb/C and C3H/SW mice except B cells. The values of lymphocyte subsets of C3H/HeJ mice were lowest among the three strains of mice. Conclusion The normal reference values of Th1/Th2 cytokine in Balb/C, C3H/HeJ and C3H/SW mice have been established. The cellular immune function of C3H/HeJ is most feeble among the three strains, and C3H/HeJ mice may be susceptible to infection with pathogenic microorganisms.

Abstract:为了有效地预防和根除山羊痘，研制能够区分自然感染和疫苗免疫的标记疫苗。本研究在山羊痘病毒（goat pox virus，GPV）疫苗株AV41胸苷激酶（thymidine kinase，TK）基因克隆、鉴定的基础上，构建转移载体。将痘苗病毒（Vaccinia Virus，VV）晚期启动子P11和大肠杆菌β-半乳糖苷酶基因（LacZ）报告基因片段插入含有TK片段同源臂的载体pTK的KpnI酶切位点，经酶切、PCR鉴定出，命名为pTK-LacZ。用脂质体法转染感染了GPVAV41株的犊牛睾丸（bovine testes，BT）细胞，待出现80%以上的细胞病变时收获病毒，经过连续9代蓝色蚀斑筛选、纯化和PCR鉴定，获得纯化的表达LacZ基因、TK功能缺失的重组病毒，命名为rGPV-LacZ。生物学特性研究显示，LacZ基因的插入不影响重组病毒的增殖特性，其毒力、生长特性与亲本株相似，保持原有疫苗株的生长特性，而且在BT细胞和羔羊睾丸（lamb testes，LT）细胞连续传20代遗传性状很稳定。本研究筛选、纯化的重组病毒rGPV-LacZ可以区分自然感染和疫苗免疫动物，为进一步研制山羊痘病毒基因工程标记疫苗奠定了基础。

Abstract:A convenient and economical method for filling DNA into microinjection capillary was described. Mouth pipette that was usually used for zygotes collection and transfer was modified for DNA filling. DNA solution was pipetted into a modified mouth pipette and then injected into microinjection capillary from the tail end. DNA filling can be finished efficiently within 2 - 5 min.

Abstract:Objective To compare and discuss the embryo transfer methods and the strains of foster mother by using 2- cell embryos from C 57 BL/6J mouse, ICR and Kunming mice as foster mother. Method Each of superovulated female C 57 BL/ 6J mice was mated individually with a stud male. The fertilized eggs were collected and cultured to 2-cell stage, then were transferred to pseudopregnant recipients. Results Using KM mice as foster mother, 2-cell embryos transferred through the wall of the fallopian tube, the pregnancy rate was 90% . The litter size was 188, rate of full-term young pups was 34.8%. Conclusion The approach of 2-cell embryos transferred through the wall of fallopian tube improve the efficiency of embryo transfer.

Abstract:Cryobiology is a jumped-up subject which investigates the influence of low temperature on organism. This extensively studied and applied technique not only promotes the progress in biology, medicine and other sciences, but also will create huge benefits for agriculture, stockbreeding, pharmacy and food manufacturing. In particular, the cryobiology has engendered far-reaching influence on environment protection, to severe endangered animal salvation and biodiversity conservation. Laboratory animals are important experimental materials for life sciences. Several scientific trends have increased the large number of mouse models and placed an intolerable load on the financial and infrastructural resources needed to maintain these models as conventional breeding colonies in recent years. The cryobiology has been applied for breeding, preservation of genetic resources, and development of new strains of laboratory animals. Anyway, the preservation of mice model will establish the furthersome condition for further studies of mammalian genetics and difficult and complicated diseases.