Abstract:目的探讨声敏剂血卟啉衍生物在肿瘤组织中的富集情况,以便在给药后超声结合血卟啉治疗肿瘤时选择最佳的超声处理时间。方法H-22肝癌荷瘤小鼠尾静脉注射HpD后,不同时间点取材,采用荧光分光光度法测定不同组织提取液中HpD的荧光强度,研究HpD在不同组织中的分布以及代谢变化。结果给药后2 h肿瘤组织以外的其他各组织内(血浆、肝、肾、皮肤、肌肉)血卟啉含量均达到其代谢过程的最高点,后逐渐下降;肿瘤组织中的HpD含量注射后不断上升,6 h时达到顶峰,随后又开始下降,10~24 h代谢比较缓慢,表现为肿瘤组织中对HpD的滞留作用,24 h时肿瘤组织中的HpD含量高于其他各组织,48~72 h趋于稳定在较低水平。结论提出了不同部位的肿瘤应选择各自适当的时间点进行超声处理。

Abstract:Objective To develop a more reliable and feasible behavioral test of experimental neurology by modifying the traditional narrow-alley test and corner test.Method\ To detect 3 groups of rats with the narrow-alley test,corner test and the narrow-alley corner test.The first group was ICH-GCSF-GROUP in which the rats suffered intracerebral hemorrhage(ICH) operation then were treated with granulocyte colony stimulating factor(GCSF).The second group was ICH-SALINE-GROUP in which the rats suffered ICH operation then were given vehicle treatment with saline;The third group was SHAM-OPERATED-GROUP in which the rats were only given a sham operation.Result(1) The same as the narrow-alley test and corner test,the rats in the SHAM-OPERATED-GROUP turned along left or right side in the same rate in the new testing apparatus,but ICH-GROUP of rats tended to turn along the operated side.(2) The testing rats need to be less stimulated in the narrow-alley corner test and the interference of people to the rats was decreased.Conclution The narrowalley-corner test is a reliable and feasible behavioral test in experimental neurology.

Abstract:Objective To improve the method in establishment of a model of renal interstitial fibrosis in rats.Methods The renal interstitial fibrosis model was established in rats by unilateral ureter obstruction(UUO).Body weight,serum creatinine(Cr) and serum urea nitrogen(BUN) level of animals from 3 groups were analyzed at the end of 1st,2nd and 3rd week.Rats were sacrificed and the pathological status of their kidneys were examined by HE,PASM and Ponceau staining at the end of 4th week.Results Cr and BUN levels in animals of the model group were significantly increased in comparison with those in sham operation group.Glomerulosclerosis,tubular atrophy,thickened glomerular/tubular basement membrane and interstitial fibrosis with inflammatory cell infiltration were observed in model group rats.A few glomerular hypertrophy and tubular dilatation were observed,too.Conclusion Model rats showing obvious renal interstitial fibrosis changes can be established by this improved technique with apparent advantage such as lower death rate,and fit for the study of renal interstitial fibrosis.

Abstract:Objective To investigate the periodical changes of hair follicles and establish animal models for studying the follicle developmental process.Method The morphology of skin hair follicles of Kunming strain mice was observed by histology during early development stage(from embryonic day 16 to postnatal day 19).Result The embryonic mouse skin epidermis had deep wrinkle,the number of wrinkles decreased on postnatal day 3 to day 5,and the wrinkles became shallow.Depth of epidermis and dermis increased stably during E16 to E19,but it reduced sharply after birth.The epidermis increased slowly stably from postnatal day 1 to day 9,the dermis increased sharply after birth.Primary hair follicles could be seen in E16 skin,the hair follicle density increased.In contrast to primary ones,secondary hair follicles appeared in E18 skin,secondary hair follicle density increased sharply and up to 1257.14/mm after postnatal day 7.The length and depth of the primary and secondary hair follicles changed a little near birth,but changed obviously from postnatal day 3 to day 7.Conclusion The changes of the mouse skin structure,primary and secondary hair follicles and the depth and length of the hair follicles are present not only in embryos but also in the early postnatal period, and in this time there is still rapid development.Mouse skin begins to change periodically.It is best to study the hair follicle gene expression in the period from E16 to postnatal day 7.

Abstract:Objective To compare the retinal vessels and especially the retinal microvessels in laboratory animals commonly used in ophthalmology and provide a basis for correctly choosing the proper animal models for ophthalmological studies.Methods The normal eyeballs of rhesus monkey,pig,rabbit,dog,cat,rat,mouse and Guinea pig were taken and the retina was peeled off.The retinal vessels were prepared with ADPase staining,examined and analyzed.Reslts The retinal vessels of rhesus monkey went from the optic disc and distributed into four branches running in four quadrants and formed a connecting net.There were two layers at the equator,one layer at the peripheral part and well-developed macular capillary ring.The vessels of pig went also from the optic disc.There was also a retinal microvascular net but no macular capillary ring.Rabbit had obvious vessels only at both sides of the optic disc and not well-developed capillaries.The vessels of dog retina were circuitous and the capillaries did not form a net.The vessels in cat,rat and mouse retinae went radially into the peripheral.The vessels in cat retina formed three branches,the superior,nasal inferior and temporal inferior.The vessels of rat and mouse did not have apparent directions.All the cat,rat and mouse retinae had well-developed capillaries consisted of two layers which were dense even at the peripheral part.Guinea pig had no distinct retinal vessels.Conclusions\ All rhesus monkey,pig,cat,rat and mouse can be used for studying the retinal vessels especially the microvessels in humans.However,only rhesus monkey can be used to study the macular retinal microvessels.

Abstract:Objective To detect and evaluate the genetic diversity and differences between SPF golden hamster population and its parent conventional population,and provide useful information and references for quality control of SPF golden hamsters.Methods Microsatellite markers of mice and rat were adopted to develop markers and methods suitable for genetic monitoring for SPF golden hamster population and its parent conventional population from Chengdu Institute of Biological Products by fluorescence-based semiautomated genotyping method.Genetic diversity and differences of the 2 golden hamster populations were analyzed based on the result of microsatellite alleles.Reslts 2 of 18 rats and 2 of 6 mice microsatellite markers amplified polymorphic loci in gold hamster.25 and 20 alleles were detected in the 4 loci,and the average of unbiased expected heterozygosity was 0.4979 and 0.5048 for conventional population and SPF population,respectively.No significant differences of genetic diversity index were found between populations.The Fst per locus was varied from 0.0095 to 0.0367,and the average Fst of all loci was 0.0315,which implied a poor genetic differentiation between populations,and most of the genetic variation existed within populations.Nei's(1972) genetic distance and Nei's(1978) unbiased genetic distance measures between the 2 populations were 0.0678 and 0.0570,respectively,which indicated the close genetic relationship and high genetic identity between them.Above half loci(2/4 and 3/4) for both populations showed significant deviations from HWE,and all deviations were caused by deficiency of heterozygous individuals by using a score test.Conclusion\ From the above mentioned results,genetic variety is mostly kept during the course of producing SPF animals in the conventional population of gold hamsters,and the genetic differences between the conventional population and SPF population are slight,but a fine propagating program may be required in the course of animal production.

Abstract:目的探讨高脂喂养联合低剂量链脲佐菌素(Streptozotocin,STZ)制备2型糖尿病大鼠模型的造模方法和影响因素。方法4周龄雄性Sprague Dawley(SD)大鼠45只随机分为三组:(1)正常组(normal control,NC),9只,普通饲料喂养。(2)高脂组(high fat,HF),9只,高脂饲料喂养。(3)糖尿病模型组,根据高脂喂养时间差异和STZ剂量不同设计了3种模型制备方法:A组,9只,高脂喂养满4周,注射STZ 30 mg/kg;B组,9只,高脂喂养满8周,注射STZ 20 mg/kg;C组,9只,高脂喂养满8周,注射STZ 30 mg/kg。所有大鼠于48h、2周和4周后行灌胃葡萄糖耐量试验(OGTT)评价成模率和血糖波动情况。实验结束时测定血清胰岛素、甘油三酯(TG)和胆固醇(TC),RT-PCR测定胰腺内胰岛素mRNA表达水平,免疫组化染色观察胰岛细胞形态学特点,用彩色图像分析系统进行定量比较。结果糖尿病C组血糖显著升高,成模后2周血糖下降,4周后又上升到基线水平,成模率100%。糖尿病A组、B组在4周后血糖逐渐降低到接近正常水平,成模率分别为55.6%、11.1%。C组与HF组相比,胰岛素敏感性显著下降(P<0.01)。β细胞内胰岛素水平下降39.3%(P<0.01),胰岛内β细胞所占比例下降了79.2%(P<0.01),胰腺内胰岛素mRNA表达水平减少19.2%(P<0.01),α细胞升高了1倍(P<0.01)。结论高脂喂养8周后腹腔注射低剂量STZ(30 mg/kg)制备的2型糖尿病大鼠模型,成模率高,模型稳定。

Abstract:目的利用原核表达系统表达仙台病毒(Sendai Virus)F蛋白主要抗原片段FP(S),并对表达产物进行免疫学初步研究。方法根据GenBank公布的仙台病毒F蛋白(gi:9627219)的基因序列设计特异性引物,通过RT-PCR扩增出F基因的主要抗原片段FP(S),插入pMD-18-T载体中,鉴定正确后克隆入pQE31原核表达载体中,将鉴定正确的pQE31-FP(S)转化大肠埃希菌M15,IPTG诱导表达,对大肠埃希菌裂解物进行SDS-PAGE和Western-blot验证。结果大肠埃希菌表达的FP(S)相对分子质量约26×103,与预期相符;能与SeV阳性血清发生特异性反应,出现单一条带。结论原核表达的FP(S)蛋白有良好的抗原性,为检测仙台病毒抗体的ELISA检测方法的研究奠定了基础。

Abstract:Objective To study the express of monkey B virus glycoprotein D in E-coli cells and detect its antigenicity.Methods BVgD was expressed in E-coli and than purified.The antigenic potential of BVgD was evaluated by Western blotting.Result The recombinant protein BVgD was successfully expressed and purified,and the result of Western blotting showed that the recombinant BVgD had good specificity and sensitivity to react with antibodies to monkey B virus.Conclusion The expressed recombinant protein has no cross reaction with HSV-1 positive serum.This protein is worthy of further development and provides valuable information for research of immunogenicity of BVgD.

Abstract:Objective To establish experimental autoimmune encephalomyelitis(EAE) models of Wistar rats and observe the expression of soluble vascular cell adhesion molecule-l(sVCAM-1).Methods Guinea pig spinal cord homogenate(GPSCH) was prepared by two methods.The first method: the whole guinea pig spinal cord homogenate was produced from the animal without PBS perfusion.The second method: the homogenate was prepared from the animal after PBS perfusion via the left cardiac ventricle.The animal model was established in Wistar rats by immunization with complete Freund's adjuvant(CFA) and GPSCH prepared by either of the two methods.Tissues samples of the brain and spinal cord were taken and paraffin sections were examined by histopathology.The expression of sVCAM-1 in animals was detected by sandwich-enzyme-linked immunosorbent assay(ELISA).Reslts The morbidity,clinical scores and expression of sVCAM-1 were significantly increased in the non-perfused group(P<0.05).Conclusion An experimental autoimmune encephalomyelitis model of Wistar rats can be successfully established with GPSCH prepared by non-perfusion method.The results of our study provided a useful and stable animal model for further studies on the pathogenesis and therapy of multiple sclerosis.

Abstract:Objective To establish a method of polymerase chain reaction(PCR) for detection of reticuloendotheliosis virus(REV).Methods The genomic DNA extracted from SPF chick embryo fibroblasts(CEFs) infected with REV-T and SNV was used as the template,followed amplification by primers from the long terminal repeats(LTR) of provirus.Consequently,the tissues from tumor-bearing chickens,as well as liver,spleen,kidney,heart,thymus and bursa from chickens on day 28 after having artificially been inoculated REV,were collected.Parts of the whole tissues were subjected to DNA extraction for PCR.The others were stained with haematoxylin and eosin (HE) and also immunohistochemical staining(IHC).Reslts Nothing amplified from REV-T was detected on agarose gel by electrophoresis.A specific and clear band sized about 300 bp was showed after amplification from SNV.Moreover,specific bands were detected by PCR of the tissues of birds infected with SNV.The tumor tissues,appearance and distribution of tumor cells were observed by histopathology(HE and histochemical staining).Conclusion The PCR is a specific and rapid technique for detection of reticuloendotheliosis virus.

Abstract:目的通过重组逆转录病毒介导获得转蚓激酶基因家兔。方法将含有蚓激酶基因的表达盒插入到逆转录病毒载体pLNCL中获得了蚓激酶重组逆转录病毒载体,利用脂质体转染PA317包装细胞,G418进行筛选得到了阳性细胞克隆,克隆扩大培养后病毒上清直接注射雄性兔的睾丸组织转染精原干细胞。结果病毒上清经NIH3T3细胞进行滴度测定,最高滴度为1×104CFU/mL。病毒注射3月龄兔睾丸组织,过1.5个月取其睾丸、肾、脾、肝、肺进行组织切片观察,结果正常。提取交配所得F0、F1代仔兔基因组,经PCR、Southern检测,转基因阳性率F0代为6.3%(2/32)、F1代为15.3%(2/13)。结论通过重组逆转录病毒直接注射3月龄兔睾丸组织可获得转基因兔,病毒对兔的脏器无损害。

Abstract:Objective To establish a small animal model of severe acute respiratory syndrome(SARS).Methods We developed a mouse model of human SARS coronavirus infection by introducing the human angiotensin converting enzyme 2(hACE2) gene,serving as the cellular receptor of SARS coronavirus(SARS-CoV),into the mouse genome.The hACE2 gene,driven by the mouse ACE2 promoter,was expressed in the lung,heart,kidney and intestines.The responses of wild-type and transgenic mice to SARS-CoV inoculation were observed.Results At 3 and 7 days after inoculation,SARS-CoV had replication more efficiently in the lungs of transgenic mice.In addition,transgenic mice showed more severe pulmonary lesions,including hyperaemia and haemorrhage in the interstitium,monocyte and lymphocyte infiltration,protein exudation,and alveolar epithelial cell proliferation and desquamation.Other pathological changes,including vasculitis,degeneration and necrosis were found in the extrapulmonary organs in transgenic mice.Viral antigen could be found in the lung and brain.Conclusions Transgenic mice are more susceptible to SARS-CoV infection than wild-type mice and the susceptibility is associated with severe pathological changes resembling human SARS infection.

Abstract:目的建立稳定的兔脑栓塞模型,以期为进一步开展脑栓塞的病理生理变化及影像学研究提供可靠实用的工具。方法30只健康新西兰兔,随机分成3组,其中A组3只,为空白对照组;B组5只,为假手术对照组;C组22只,为栓塞组。分离右侧颈部血管,经颈外动脉向颈内动脉注入直径约0.5~1.0 mm的SiO2颗粒10枚左右,栓塞后30 min行CT灌注检查,利用多层螺旋CT灌注成像对各组动物的脑缺血情况进行观察。24 h处死动物取脑组织进行病理研究。结果A、B组CT灌注及病理均未见异常。C组栓塞过程中3只兔死亡。16只兔CT灌注异常,表现为右侧局部CBF降低、MTT延长、CBV无明显变化或轻度上升、下降。3只兔灌注未见异常。HE染色可见8只兔脑梗塞,7只兔脑缺血,4只兔未见明显异常。结论采用该方法和技术能够建立稳定的兔脑栓塞模型,结果可靠,具有可操作性和重复性。

Abstract:目的建立猪心脏移植供体的急性脑死亡模型,观察急性脑死亡前及后1、5、10 min时的血流动力学和血浆中儿茶酚胺释放的变化并进行初步探讨。方法采用30~40 kg的猪8只,急性脑死亡前为对照组,急性脑死亡后为实验组。测定脑死亡前基础水平及脑死亡后1、5、10 min时血流动力学改变和血浆中儿茶酚胺释放的变化,并对二者的变化进行初步探讨。结果急性脑死亡后1 min时血流动力学及儿茶酚胺水平改变最明显,心率增加了88%,收缩压升高了132%,心排量增加了80%,肾上腺素(E)和去甲肾上腺素(NE)分别升高240%和241%,多巴胺(DA)没有明显增加。随后二者均持续下降,至10 min时已降至基础水平以下。血流动力学改变的程度及时间分布与儿茶酚胺中的肾上腺素和去甲肾上腺素一致。结论急性脑死亡可造成机体血流动力学及血浆儿茶酚胺水平的剧烈改变,血中儿茶酚胺含量升高是造成急性脑死亡后血流动力学改变的原因,其中E和NE与血流动力学改变直接相关。

Abstract:As one of the four model animals,zebrafish(Danio rerio) has been widely used in researches on embryology,developmental biology,reproductive toxicology and molecular biology.But merely a handful of studies have been reported on its reproductive endocrinology and environmental toxicology.This paper summarized the development of studies on reproductive endocrinology in zebrafish as a model animal.

Abstract:Rheumatoid arthritis(RA) is a common disease that causes deformities,but its pathology is not fully clear.Establishment and application of the animal models is an effective method for RA research.In this article the methodology,mechanisms and applications of current animal models of RA were reviewed,their characteristics and drawbacks were analyzed,and prospective directions of rodent animal models of RA were put forward.