Abstract:第一期禽流感病毒A型、H5亚型RT-PCR检测试剂盒研究孙明,李纯铃,赵铁柱等(3)…………………………………禽流感病毒H7N2血凝素HA1基因在大肠杆菌中的表达多海刚,冉多良,孙明等(7)………………………………禽流感病毒分离株NS基因同源性及等位基因类型分析王传彬,孙明,赵铁柱等(11)………………………………Cx43基因在人类及小鼠胎心发育中的时空表达规律陈萍,黄国英,常才等(16)……………………………………6-羟多巴胺脑内注射制备帕金森病大鼠模型的研究范凯,马坚妹,马晓凯(20)………………………………………p16真核表达载体的构建…

Abstract:Objective Members of the epidermal growth factor (EGF) family of ligands and their receptors regulate migration and growth of intestinal epithelial cells: integrins, the immunoglobulin superfamily, selectins and cadherins. Each family is composed of a number of constitutional members. These cell adhesion molecules mediate the process of cellular adhesion to either the extracellular matrix (ECM) or initiate cell-cell adhesion. Focal adhesion kinase (FAK) is a major mediator of integrin signaling pathway. In this paper, the interaction between FAK and EGFR, and the regulation of FAK on the EGFR signaling pathway was studied. Methods Cell lines expressing del 1-693FAK-GFP, del 1-100FAK-GFP and FAK-GFP were used. The deletion mutants of FAK, del 1-693FAK-GFP and del 1-100FAK-GFP lost the ability to recruitment to adhesion sites. PI electrophoresis and Western blot analysis were applied. Results PI electrophoresis analysis indicated that the phosphorylation of FAK was regulated by EGF stimulation. Western blot analysis showed that expression of del 1-693FAK-GFP and del 1-100FAK-GFP inhibited the phosphorylation of MAPK. However, FAK-GFP enhanced the phosphorylation of MAPK and inhibited the phosphorylation of Akt. Conclusion The results suggested that FAK regulates the balance between MAPK and Akt signal pathways through interaction with EGFR.

Abstract:目的建立人乳腺癌MCF-7细胞SCID(Severe combined immunodeficiency,SCID)小鼠转移动物模型。方法采用人乳腺癌细胞株MCF-7细胞悬液,分别接种于5只经放射线处理的SCID小鼠腋背部皮下。记录肿瘤生长情况,处死荷瘤鼠并做病理切片,观察各脏器转移情况。结果接种SCID小鼠后6～10d成瘤,成瘤率为5/5只,潜伏期平均(7.4±1.3)d。接种后5只鼠分别于第60～68天拉颈处死,检测荷瘤,平均直径为(26.6±2.2)mm,平均重量为5.28g。病理学检查,转移脏器有3个部位,出现肺转移的为4/5只、骨转移的为3/5只和淋巴结转移的为1/5只。结论建立了人乳腺癌SCID小鼠转移动物模型,该模型可为肿瘤转移研究提供重要的实验工具。

Abstract:目的研究纳米珍珠粉对大鼠钙吸收利用的影响,并比较纳米和微米珍珠粉对大鼠生长发育和骨钙水平作用的差异。方法出生21d断乳SD大鼠100只,喂养低钙饲料2周。随机分为5组,雌雄各半,Ⅰ～Ⅱ组分别为微米珍珠粉低、高剂量组,喂饲饲料为低钙饲料混合不同剂量的微米珍珠粉;Ⅲ～Ⅳ分别为纳米珍珠粉低、高剂量组,喂饲饲料为低钙饲料混合不同剂量的纳米珍珠粉;Ⅴ组为低钙对照组,喂饲低钙饲料;另取10只SD大鼠为实验本底组(即Ⅵ组),Ⅵ组于实验开始前处死。实验期为4周。结果2个不同剂量的纳米珍珠粉组的大鼠体重、股骨干重及长度、钙吸收率、钙存留率均高于2个相应剂量的微米珍珠粉组,且差异有显著性(P<0.05)。结论纳米珍珠粉能够被大鼠很好地吸收利用,具有增加骨钙含量的功能,在钙吸收利用作用方面明显优于微米珍珠粉。

Abstract:Objective This study tried to reduce the observation time points in the establishment of a murine menstruation model, which was originally developed by Finn CA in 1984. Therefore, to make the murine menstruation model more economical and easier to handle. Methods Adult female C57BL/6 mice were ovariectomized, then sequentially treated with hormones. 4-6 hours after the last hormone injection, 0.02 ml sterilized arachis oil was injected into the uterine lumen of experimental mice. The control group only received the sham operation. Uteri were collected after oil treatment for 31-35 hours (group T3) and 56-70 hours (group T4), measuring wet uterine weight, doing H&E staining and using CAST2 image analysis software to measure the total uterine cross-sectional area (TUA) and endometrial cross-sectional area (EA). Results Through histological examination, estrogen treated group showed simple columnar epithelial cells, with high nucleus-to-cytoplasm ratio and elongate sausage-shaped nuclei with dense chromatin. After combined treatment with progesterone, secretory cells and their secretions were easily found in the glandular lumens. After oil treatment endometrium sloughing was observed in group T3, while endometrial epithelium repaired completely in group T4. Endometrium of control groups kept intact during the whole process. Wet uterine weight had no significant changes during sequential hormone treatment. After hormone withdrawal, the wet uterine weight decreased in the experimental groups slower than that of control mice. TUA and EA significantly increased after combined treatment with progesterone. 30-35 hours after oil treatment TUA increased, whereas EA did not changed. 56-70 hours later both of them decreased. Conclusion The histological changes of endometrium in this murine menstruation model displayed many features resembling human menstruation, especially, the endometrial subluminal epithelium sloughing and early endometrial repair.

Abstract:目的评价Dextran不连续密度梯度离心法纯化大鼠胰岛的效果。方法采用V型胶原酶分离出大鼠胰岛,并应用Dextran不连续密度梯度离心法纯化胰岛。在体视镜下计数双硫腙染色的胰岛并测量染色胰岛的直径。放免法测定胰岛素含量。AO-PI双染色确定胰岛的活力。结果平均每只成年Wistar大鼠的胰腺可分离950±24个胰岛,经Dextran纯化后平均每只成年Wistar大鼠的胰腺可获得784±10个胰岛,纯度可达到90%以上。结论采用Dextran不连续密度梯度纯化得到的Wistar大鼠胰岛结构完整、功能良好。

Abstract:Objective Banna Mini-pig Inbred Line 133 family(BMI-133) is one of the sub-lines which is differentiated from the Banna Mini-pig Inbred Line(BMI) with stable genetics and high homozygosity. It has been found that BMI-133 has many similarities with human such as organ anatomy, physiology and biochemcal indexes and so on. The purpose of this study is to explore the potential application of BMI-133 in xeontransplantion for treatment of human diseases. Method The DQ genes of BMI-133 were cloned and sequenced by RNA extraction, reverse transcription, cDNA amplification and transformation. Two complete open reading frames (ORF) have been obtained. Result The cDNA of DQA BMI-133 has 830 nucleotides and that of DQB has 1103 nucleotides. They encode 255 and 262 amino acids, respectively. Conclusion The results showed that BMI-133 has distinct differences with other mini-pigs reported, they are two new alleles. They also have high similarity to the counterpart sequences of human. It is expected that following further research on BMI-133, it may become a most suitable animal for xeontransplatation.

Abstract:Objective To explore the pathogenesis of steroid-induced osteonecrosis in serum disease animal. Methods New Zealand rabbits were distributed into three groups randomly. Horse serum and dexamethasone sodium phosphate were used to induce osteonecrosis models in group A, dexamethasone sodium phosphate only in group B and saline in group C. To test and analyze the values of serum lipid, AST and ALT of all rabbits. Rabbits from all groups were observed by MRI to prove osteonecrosis and sacrificed to observe the histopathology of femoral head. Results Typical avascular necrosis of the femoral head happened in group A. The values of serum lipid, AST and ALT are distinctly different from those of other groups of which rabbits did not develop osteonecrosis. Conclusion Horse serum combining dexamethasone sodium phosphate can induce rabbit osteonecrosis model successfully. Vasculitis, abnormal serum lipid metabolism and liver disfunction together result in avascular necrosis of the femoral head. High dose steroid can not induce osteonecrosis in a short time.

Abstract:目的观察肿瘤坏死因子α(TNFα)对正常昆明小鼠骨骼肌和肝脏葡萄糖摄取的影响。方法80只昆明小鼠随机分为高剂量连续注射组(TH组,n=20)、TNFα低剂量连续注射组(TL组,n=20)、TNFα高剂量一次注射组(T1组,n=20)和正常对照组(C组,n=20),以3H标记的2脱氧葡萄糖(2-DG)为示踪剂,观察各组非胰岛素刺激(基础)和胰岛素刺激的离体骨骼肌和肝脏葡萄糖摄取量的变化。结果1)正常昆明小鼠肝脏基础葡萄糖摄取量明显高于骨骼肌葡萄糖摄取量(P<0.01),肝脏胰岛素刺激的葡萄糖摄取量变化值明显低于骨骼肌葡萄糖摄取量变化值(P<0.01)。2)TH组和TL组骨骼肌和肝脏基础葡萄糖摄取量均明显高于C组(P<0.01),且TH组明显高于TL组(P<0.01)。3)TH组和TL组骨骼肌和肝脏胰岛素刺激的葡萄糖摄取量均明显低于C组(P<0.01),且TH组明显低于TL组(P<0.01)。4)T1组骨骼肌和肝脏无论基础还是胰岛素刺激的葡萄糖摄取量均与C组差异无显著性(P>0.05)。结论1)正常动物肝脏和骨骼肌葡萄糖摄取方式不同,骨骼肌葡萄糖摄取受胰岛素调控比肝脏强。2)TNFα抑制组织胰岛素刺激的葡萄糖摄取,但促进组织基础葡萄糖摄取。3)TNFα对组织葡萄糖摄取的影响,呈剂量和时间依赖性。

Abstract:目的获得泰泽氏病原体抗原表位相关肽,用于实验动物血清中该病原体感染相关抗体的检测。方法选用泰泽氏病原体的四种单克隆抗体(M2、M3、M4、M5)作为配基,从噬菌体表面展示的随机7肽文库中筛选单抗识别的抗原表位,获得特异性噬菌体克隆;并采用ELISA、Western blot方法对其进行分析鉴定,获得阳性噬菌体克隆。结果获得阳性噬菌体克隆5个,其展示的融合蛋白能被泰泽氏病原体的免疫血清识别,ELISA检测A值的P/N为8.0～17.1;Western blot分析显示单一特异性条带,相对分子质量约为38×103。结论本研究获得的5个阳性克隆所表达的融合蛋白,为泰泽氏病原体抗原表位相关肽,可作为该病原体隐性感染血清学检测的候选抗原。

Abstract:Objective Previous study showed that one of several quantitative trait loci (QTL) responsible for weight of spinal cord in mice located at D15Mit158 with an interval of 30 cM on chromosome 15. To identify related genes, fine mapping of QTL within this region was conducted in this study. Methods An advanced intercross line (AIL) from A/J C- 57 BL/6J (F4) mice was used for partial genome scanning. High-resolution microsatellite markers around D15Mit158 were used to genotype in the F4 mouse population representing phenotypic extremes. Linkage analysis was performed by software Map Manager QTX19. Results A peak LRS-value of 17.3 was identified around D15Mit107 (p =1.8 10 -4 ), its contribution to the variance was 24% with an interval of 3.2 cM, and the LOD score was 3.75. Another suggestive linkage site was found around D15Mit28, its LRS-valve was 7.6(p =0.02) and contribution to the variance was 13%. Conclusion The QTL region of D15Mit158 contained two QTLs actually after fine mapping. One major QTL was around D15Mit107 with an interval of 3.2 cm, and another suggestive QTL was around D15Mit28 with an interval of 5 cm.

Abstract:Objective In order to set up particular mutation mice population of SPF by using purging method. Methods Morulae of four mutation hairless mice and one cataract mouse were transplanted into uterus using embryonic transfer method. Results The embryonic transfer was succeeded with birth of infant mice of the four mutation hairless mice and one cataract mouse. The highest rate of success was 25.5% in the birth of cataract mice. Conclusion Particular mutation mice population of SPF has been established, which may provide a reliable support for various research studies and further development utilization.;

Abstract:目的研究FMMU白化豚鼠的mtDNA,并与花色豚鼠mtDNA进行多态性分析比较,以确定其独特的生物学特性是否与mtDNA相关。方法用碱变性法提取FMMU白化豚鼠以及花色豚鼠的mtDNA,并用AvaⅠ、BalⅠ等12种限制性内切酶进行酶切和限制性片段长度多态性分析。结果与结论FMMU白化豚鼠mtDNA和花色豚鼠mtDNA的相对分子质量相同,约为16.7×103;FMMU白化豚鼠与花色豚鼠两品系的mtDNA经AvaⅠ、BalⅠ等内切酶酶切后有3-8个酶切位点,酶切图谱完全相同,经RFLP分析FMMU白化豚鼠与花色豚鼠的mtDNA之间缺乏多态性。本实验没有发现FMMU白化豚鼠的独特的生物学特性与mtDNA相关。

Abstract:Objective To make measurement of the estradiol (E-2) and progesterone (P) level in blood serum of Mongolian gerbils, and to compare with those in NIH mice and SD rats. Method The serum E2 and P of these laboratory animals were measured by RIA and statistically analyzed. Results The serum level of E-2 in virginal and multiparous Mongolian gerbils is not significantly changed, and serum P is higher in multiparous gerbils (P<0.01). There is a significant difference between the E-2 level in virginal Mongolian gerbils and in NIH mice and SD rats (P<0.01), but similar in multiparous gerbils (P>0.05). The P level in virginal gerbils is similar to that of NIH mice, but significantly different with that in SD rats (P<0.01). The serum P level in multiparous gerbils is highest in comparison with that of NIH mice and SD rats (P<0.01). Conclusion The estradiol and progesterone levels in blood serum are species specific in Mongolian gerbils and change with the growth and development of the animals.

Abstract:目的建立以重组外膜蛋白为基础的钩端螺旋体抗体间接ELISA检测方法。方法以基因重组技术获取重组钩端螺旋体外膜蛋白LipL32,以该蛋白为抗原,特异的钩体抗血清进行ELISA方阵滴定、交叉性试验、阻断试验,并对北京地区的70份犬血清使用建立的ELISA方法以及德国Virion公司的全菌体钩端螺旋体ELISA试剂盒进行相互验证。结果方阵滴定试验确立以100ng/孔为抗原包被浓度,1∶160为血清稀释度。交叉性试验具有广泛性、阻断试验标明该方法特异性强、灵敏度高。两种方法数据经χ2检验,两者检出率之间差异不显著。结论重组LipL32蛋白具有结合活性。初步建立了以重组LipL32蛋白为抗原的钩端螺旋体抗体间接ELISA检测方法。

Abstract:Objective To investigate the protective action of oxygen-breathing pretreatment against brain ischemia-reperfusion injury in rats. Methods Using a model of focal brain ischemia-reperfusion in Sprague-Dowley rats, SOD activity and MDA level measurement, electron microscopy and neurological functional examination were performed to evaluate the protective role of oxygen-breathing pretreatment. Results Oxygen-breathing pretreatment showed a protective effect on brain ischemia-reperfusion injury. It presented significantly increased SOD level and decreased content of MDA. The ultrastructure of neurons in rats of oxygen-breathing pretreated group was ameliorated effectively. Conclusions Oxygen-breathing pretreatment has protective effect against brain ischemia-reperfusion injury.

Abstract:Objective Human papillomaviruses (HPVs) are associated with specific benign and malignant lesions, such as cervical carcinoma, anogenital cancers, condyloma acuminata, etc. However, they are human epithelia-specific and do not infect any laboratory animals. This characteristic makes it difficult to study the infection process and pathogenisis of HPVs, and to develop anti-HPV drugs. To solve these problems, it seems important and urgent to generate animal models with HPV. Thus a review of advances in this field was made in this article.

Abstract:1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced animal models are ideal models of human Parkinson's disease, in terms of neurobiochemical and histopathological changes, as well as behavioral disorders. Intensive research into the pathogenesis and other fields of MPTP-induced lesions in the animal models will contribute much to the prevention and cure of human Parkinson's disease.