为探究低温下香樟叶肉含晶细胞和溶酶体的变化规律,以三年生香樟为试验材料,以常温为对照,通过人工模拟低温,分别于5和O℃条件下处理48 h,经制样切片后,用透射电子显微镜观察叶肉含晶细胞和溶酶体的变化.结果表明:常温处理下,香樟叶肉部分细胞中晶体数量多、体积大,晶体的形态多为柱状晶和方晶;5℃低温处理下,液泡中晶体的数量较常温略有减少,晶体体积也略有变小,晶体形态以柱状晶和方晶为主;0℃低温处理下,细胞液泡内晶体的数量急剧减少,晶体的形态以针晶为主,伴之少许方晶.不同温度处理的香樟叶肉细胞液泡内均有溶酶体,随着处理温度的降低,观察视野内单个细胞中溶酶体的数量略有增加.

为了研究膨化血粉对黄河鲤鱼生长性能和肠道形态的影响,试验采用体重为(12.82±0.30)g的黄河鲤鱼120尾,随机分为两组,分别以含17％进口秘鲁鱼粉的基础饲粮(对照组)和以膨化血粉等比例全部替代鱼粉的试验饲粮(试验组)饲喂7周.结果表明:试验组鱼的最终体长,最终体重,增重率,前、中、后肠的绒毛高度和密度,前肠和后肠的绒毛宽度均极显著低于对照组(P＜0.01),饲料系数极显著大于对照组(P＜0.01);大部分肠上皮细胞纹状缘消失,部分前肠、大部分后肠上皮细胞呈现空泡变性,部分中、后肠绒毛顶端上皮细胞坏死、脱落.说明饲粮中膨化血粉全部替代鱼粉的比例达到17％会显著降低黄河鲤鱼的生长性能,减少肠上皮细胞数量,破坏肠绒毛结构.

To study the cerebral granulomatous formation caused by Encephalitozoon cuniculi(E.cuniculi)through ERK signal transduction pathway,the common staining,special staining,immunohistochemistry and Western-blot were used to observe the morphological changes of cerebral granuloma by E.cuniculi.Moreover,analyze the relationship between the formation of cerebral granuloma and four kinds of key proteins,such as NGF,TPK,MAPK and ERR in ERK signal transduction pathway was investigated.The results showed that there were a number of cerebral granuloma and nonsuppurative encephalitis in all of sick rabbits.The expression of NGF,TPK,MAPK and ERK was obviously higher in astrocytes,vascular endothelia and fibroblasts of the cerebral granuloma.These cells showed strong positive reaction and were stained as dark brown by immunohistochemistry staining.The key protein abundance of signal transduction pathway in sick rabbit's brain tissue was much higher than that in control rabbit's by Western-blot.It was indicated that the ERK signal transduction pathway was activated in the process of forming cerebral granuloma by E.cuniculi and could facilitated the proliferation of astrocytes,endothlia and fibroblast formed cerebral granuloma via strong cascade reactions.

To study the relationship between cerebral granulomatous formation by Encephalitozoon cuniculi and PLCγ/PKCsignal transduction pathway and expression of symbolic proteinglial fibrillary acidic protein in astrocytes,the common staining,special staining,immunohistochemistry and Western-blot were used to observe the morphological changes of cerebral gramuloma by E.cuniculi and analyse the relationship between the formation of cerebral gramuloma and the key proteins of PLCγ/PKC signal transduction pathway,andGFAPexpression in the proliferative astrocytes.The results showed that there were a number of cerebral granuloma and nonsuppurative encephalitis in all sick rabbits.The expression of FGF,PLCγ,P-PLCγ and PKC was obviously higher in astrocytes,vascular endothelia and fibroblasts of the cerebral granuloma.These cells showed strong positive reaction and were stained as dark brown by immnohistochemistry staining.GFAPonly expressed in astrocytes,especially in proliferative astrocytes.The key protein abundance of signal transduction pathway in sick rabbit's brain tissue was much higher than that of the control by Western-blot.GFAP expression of sick rabbits'brain tissues was apparently higher than that of the control.The conclusion is that the PLCγ/PKC signal transduction pathway is activated in process of forming cerebral granuolma by E.cuniculi via powerful cascade reaction.GFAP expression in astrocytes is a proliferative basis of astrocytes in the cerebral granuloma,and is also a manifestation of proliferative genes activated.

[目的]建立一种利用CMV启动子获得拯救猪繁殖与呼吸综合征病毒(PRRSV)的方法,为进一步研究猪繁殖与呼吸综合征病毒提供操作平台.[方法]根据PRRSV的基因序列特征和CMV启动子的序列特征,对pBluescprictⅡSK(+)载体进行多克隆序列改造;设计特异性引物,分6段扩增、组装出PRRSV的全长cDNA并扩增CMV真核启动子序列;采用酶切、连接、转化等方法将连接成功病毒的全长cDNA置于CMV真核启动子序列的下游,获得含有CMV启动子和猪繁殖与呼吸综合征病毒CH-1A株全长cDNA克隆的重组质粒pSK-CH-1A;再将重组质粒直接转染MARC-145宿主细胞,得到猪繁殖与呼吸综合征病毒CH-1A株病毒的拯救病毒.[结果]成功获得了19 112 bp含CH-1A全长cDNA的pSK-CH-1A载体,将其直接转染MARC-145宿主细胞后,经细胞体内转录合成病毒基因组RNA,获得了拯救病毒(rCH-1A);病毒生长特性试验结果显示,拯救病毒与亲本病毒在MARC-145细胞上具有相似的增殖特性,且拯救病毒序列含有不同于亲本病毒的分子标记MluⅠ酶切位点.[结论]建立了一种方法简单,操作方便,节约时间和成本的PRRSV感染性cDNA克隆的体外拯救方法.

152 AA+ male broilers at four-day-old were randomly divided into four treatments and fed on diets with different levels of thiamphenicol such as 0 mg/kg (control),500 mg/kg(group Ⅰ),1 000 mg/kg(group Ⅱ),and 1 500 mg/kg (group Ⅲ) for 2 weeks for studying the poisoning effect of thiamphenicol on testis morphology in broolers. After 2 weeks,the body weight,testis weight,testis organ coefficient,seminiferous tubule area and some pathological change of the testis were measured and observed. The results showed that the body weight and seminiferous tubule area of three treated groups and the testis weight of group Ⅱ and Ⅲ were significant lower than those of control groups (P0.01). The testis weight and testis organ coefficients of group Ⅰ were significant higher than those of control groups (P0.01). The germ cells in the seminiferous tubules of three treated groups decreased at different degrees. The spermatogonia and sertoli cells were observed blister degeneration or even necrosis and desmoplastic. It indicated that the testis of broilers might be injured when fed with thiamphenicol above 500 mg/kg weight,and the damage degrees maybe correlated to thiamphenicol dosage in a certain extent.

免疫接种是控制传染病的重要手段．但在动物生产实际中经常遇到接种过疫苗后仍然发病的现象．这就是通常所说的免疫失败．给畜禽生产造成重大经济损失。笔者根据实践经验及查阅相关文献，对引起畜禽免疫失败的主要因素总结如下．以供参考。

The objective of this study was to evaluate the effects of different level of expanded blood meal in the diet on the serum biochemical indexs and serum hormone of goose.300 Huoyan geese(78.68±2.95 g BW;28 days of age) were randomly divided into three treatments with five replicates.The gradients of expanded blood meal level in the formulated diet were 1.5%,3.0% and 4.5% respectively.The results were as follows:serum AST and ALB in 3.0% group were significantly higher than that of 4.5% group(P0.05),but there was no significant difference between the two groups mentioned above and 1.5% group(P0.05).The difference of serum Glu between 3% group and 4.5% group was not significant(P0.05),but serum Glu of the two group mentioned above were significantly higher than that of 1.5% group(P0.05).Serum TSH and T3 in 3.0% group were significantly higher than that of 1.5% group and 4.5% group(P0.05),but there was no significant difference between 1.5% group and 1.5% group(P0.05).The difference of ALT,GLOB,TP,INS,GH,T4 and IGF-1 in all the three groups were not signifcant(P0.05).

The PCR products of length of 414 bp and 450 bp were derived from air of new cell culture room by using random PCR,and were cloned into pGEM-T vector for sequencing and analysis.Results showed that the identity of the nucleotide sequence of obtained genes between HN09 and HN10 strains and other Cupriavidus metallidurans reference strains were from 79%-83% and 79%-84%,respectively,and the identity of the deduced amino acid sequences of obtained genes between HN09,HN10 strains and other Cupriavidus metallidurans reference strains were from 86.4%-89.1%.The present study indicated that these two strains may belong to different subtypes of Cupriavidus metallidurans.

The article discusses operation instruction,announcements and daily maintenance of J-25 console model high speed refrigeration centrifuge and L-100K ultracentrifuge.It would offer useful references to administrators and users for educing centrifugal maximum efficiency.It would meet relevant requirements of professional experimenters for animal science,food science and life science of Henan Institute of Science and Technology,and would enhance teaching and research level of comprehensive university to make some contributions.

Veterinary pathology,which is highly theoretical and practical,is an important public basic course of animal medical specialty or relevant.In response to the features above,the following teaching methods have been taken in the teaching of veterinary pathology in order to excite the students' interest in veterinary pathology study and improve the quality of teaching.Firstly,teaching is combined with self-study to arouse student's enthusiasm in learning.Secondly, a heuristic approach and mass means of instructions have been taken in teaching. Thirdly,the task assigned by teachers is required to be rational.Fourthly,theory must be integrated with practice in the teaching process of veterinary pathology.In addition,the exam should be carried out in many ways.

微粒体甘油三酯转运蛋白(MTP)是继载脂蛋白B(apoB)之后发现参与TG转运及VLDL组装的内质网腔内蛋白,是一种重要的脂质转运蛋白.MTP主要作为载体在内质网腔与含脂蛋白的apoB等脂类结合以VLDL的形式转运出肝脏,对含脂蛋白的apoB装配和分泌起关键作用[1].

Primary culture is defined as simulating the in vivo physiological environment to make the cells survival,growth and passage in vitro,and maintain the original structure and function of cells.Chick embryo fibroblasts have characteristics,such as easy obtaining,high proliferation ability and adaptability,good tolerance,relatively stableness,making them widely used in molecular and cytobiologic research in many ways.The article reviewed the current status of chicken embryo fibroblast culture techniques,such as the selection of embryo,cell separation,purification and culture,and also introduced the application and research directions in the future.

The harmful effect of dietary thiamphenicol on the pathology of immune organ in the chicken was investiga-ted. 304 four-day-old AA+ chickens were randomly divided into four groups,and fed on diets as follows: 0,500, 1 000 and 1 500 mg/kg for 6 weeks. Clinical signs and mortality were observed daily and body weight was measured weekly during the treatment. After 2,4 and 6 weeks, five of'three treated groups and control group were examined for the amounts of RBC, hemoglobin level,immune organs ralative weight and histology of the immune organs by paraffin section. The mortality were 0 %,9.21%, 64.47 % and 68.42 %, respectively. Appetite of chickens receiving TAP decreased,and growth rate was much retarded significantly. The Hb level and the RBC amounts dropped. Or-gans ralative weight of bursa of Fabricius and spleen and thymus decreased; leukomonocyte of the immune organs in the chicken decreased. The results demonstrated that 500 mg/kg thiamphenicol in diet could induce morphological in-jury of immune organ especially humoral immune organ, and damage degrees may be relative to thiamphenicol dosage in a certain extent.

Veterinary pathology,as a highly theoretical and practical course,is an important public basic course of Animal Medical Specialty.In response to the features above,the following teaching methods should be taken in the teaching of Veterinary Pathology in order to raise the study interest ofstudents and improve the teaching quality,such as the combination with self-study,heuristic mode of teaching,mass means of instructions,task assigned by teacher,the integration of theory and practice,and the reform of exam forms.

The laboratory centrifuge in China especially high speed refrigeration centrifuge and ultracentrifuge have been improving rapidly in recent years. But the articles on principle, manipulation, announcements and maintenance of centrifuge were seldom. In this article, we overviewed the key techniques on principle, manipulation, announcements and maintenance of centrifuge. It would offer useful references to administrators and users for educing centrifugal maximum effi ciency.

The effect of dietary thiamphenicol on the haematological indexes in chicken was investigated.Three hundred and four four-day-old AA+ chickens were randomly divided into four groups,and fed on diets as follows :0 mg/kg(control),500 mg/kg(group Ⅰ),1 000 mg/kg(group Ⅱ) and 1 500 mg/kg(group Ⅲ) for 8 weeks.After 2,4,6 and 8 week,five of three treated groups and control group were examined for the amounts of RBC,and level of hemoglobin,TP,ALB,ALT,AST,BUN,Crea and Fe.The results were as follows:The Hb level and the RBC amounts were decreased significantly(P0.05);TP and ALB were increased in the early stage and decreased significantly in the later stage(P0.05);serum AST were decreased significantly(P0.05);serum Crea and BUN were increased significantly(P0.01);serum Fe were increased significantly(P0.05).The results demonstrated that over 500 mg/kg thiamphenicol in diet could impact the significant changes of haematological indexes to a certain,extent with dose-response.

4日龄AA+肉鸡304只,随机分为4组,分别以0,500,1 000,1 500 mg/kg饲料的甲砜霉素剂量饲喂6周,研究甲砜霉素对肉鸡免疫器官的病理学损害.试验开始后每天观察肉鸡的临床症状,每周称重1次,计算平均体重,分别在第2,4,6周测定红细胞数量、血红蛋白含量、免疫器官脏器系数,并取免疫器官作石蜡切片以研究甲砜霉素对肉鸡免疫器官的病理组织学影响.结果发现,各组死亡率分别为0%,9.21%,64.47%,68.42%;中毒鸡表现为食欲减退、体重增长缓慢、贫血;法氏囊、脾脏和胸腺脏器系数显著降低,淋巴细胞减少.上述研究结果表明,即使以500 mg/kg饲料的甲砜霉素持续饲喂肉鸡,均可对肉鸡的免疫器官结构造成明显损伤.尤其是体液免疫器官,并且这种损伤存在剂量效应关系.

We determined the contents of dihydromyricetin in Ampelopsis grossedentata leaves from different sources by HPLC, and evaluated the antioxidant and cytotoxic activity of dihydromyricetin. Ampelopsis grossedentata leaves were rich in dihydromyricetin. Dihydromyricetin was an effective scavenger in quenching 1, 1-diphenyt-2-picrylhydrazyl radical with IC50 of 21.48 mu M. In the Rancimat antioxidant test with lard oil as substrate, dihydromyricetin had a protection factor of 11.29, superior to that of tertiary butyhydroquinone (protection factor of 4.75). In the cytotoxic assay, dihydromyricetin inhibited the growth of human epidermal cancer A431 cells in a concentration-dependent manner.

针对我院提高办学层次、购进仪器设备、建设实验中心科研平台的契机,应转变思想观念,以建立和完善仪器设备管理模式,发挥实验室管理人员在实验室中的主导作用,教学和科研双管齐下,建设出一个管理规范、高效的实验中心.