鸡海马结构传入纤维的端脑起始核DiI法研究

研究鸡海马结构传入纤维在端脑内的起始核。采用DiI逆行追踪法,将DiI注入鸡端脑海马结构,逆行追踪投射到海马结构的起始核。结果在端脑的副高纹状体、内侧隔核、古纹状体、带核和后背外侧区出现标记神经元。初步表明端脑内的上述核团是向海马结构投射的起始核。     

鸡端脑向小脑投射的起始核

将40%辣根过氧化物酶（HRP）水溶液注入鸡小脑的Ⅵ、Ⅶ、Ⅷ叶，逆行追踪其端脑向小脑投射的起始核。结果发现，端脑的海马列、隔核、带核、旁嗅叶出现多量标记细胞；副高纹状体、嗅结节和古纹状体出现少量标记细胞。将CB-HRP引入古纹状体，于小脑皮层的分子层内出现顺行标记物，颗粒层内也有稀疏的顺行标记物出现。结果表明，鸡端脑-小脑投射的起源较为丰富而广泛，上世纪初提出的纹-小脑束可能起始于古纹状和带核。     

鸡海马结构传入纤维的脑干及小脑起始核—HRP逆行示踪法

研究鸡海马结构的传入纤维在脑干及小脑内的起始核。将40%辣根过氧化物酶(HRP)水溶液注入鸡左端脑海马结构,逆行追踪投射到海马结构的纤维在脑干和小脑内的起始核。结果在脑干的浅小细胞核(SPC)、圆下核(SRt)、蔡氏区(AVT)和小脑的第Ⅰ~Ⅹ叶浦肯野细胞层内均出现被标记的神经元。初步表明上述部位为端脑海马结构的起始核。     

鸡端脑和下丘脑向面神经腹侧核的传出性纤维联系—HRP逆行追踪法

将２０％ＨＲＰ水溶液注射于鸡的面神经腹侧核，逆行追踪支配该部的有关主同位神经核团。结果于端，下丘脑和脑干内出现标记细胞，端脑内的标记细胞较多地出现于古纹状体，少量出现于旧纹状体腹侧与枕中脑束之间的区域，两群标记细胞形态上明显不同。下丘脑的标记细胞较多地出现于下丘脑后区，室旁核出现少量，个别散在于下丘脑其他区域。     

鸡上纹状体的纤维联系——HRP法研究

将HRP溶液注入鸡上纹状体,追踪投射至上纹状体的纤维来源,在丘脑背外侧前核、背外侧后核,丘脑背内侧前核、背内侧后核,中脑深核外侧部和腹侧部及顶盖背核出现标记细胞。外侧前脑束和隔中脑束出现标记纤维。结果表示.上述核团有纤维投射到上纹状体,且投射均限于同侧。     

鸡海马结构向小脑各叶的直接投射——HRP逆行追踪法研究

采用HRP法逆行追踪鸡海马结构向小脑各叶投射的起始神经元.将50%HRP溶液分别引入鸡小脑Ⅳ、Ⅴ、Ⅵ、Ⅶ、Ⅷ、Ⅸ各叶,对端脑及间脑、脑干及小脑进行冰冻切片,TMB呈色,观察脑内标记细胞出现的位置.结果发现,除第Ⅸ叶外,注射小脑各叶双侧端脑海马结构的旁海马内侧区(APHm)出现大量标记细胞.在注射Ⅳ、Ⅴ、Ⅵ后偶尔在内侧隔核出现少量逆标细胞.随着注射点由Ⅳ叶到Ⅸ叶的后移,在APHm出现标记细胞的可能性渐小,而小脑前核,包括延髓大细胞网状核、下橄榄核、内外侧桥核、中脑的内侧螺旋核等标记的可能性相应地增大.隔核的标记细胞主要定位于外侧隔核.本研究结果表明,鸡存在海马向小脑的直接投射,这种投射主要终止于小脑的前叶、中叶.因为哺乳动物,鸡的隔核与海马有密切的联系.     

鸡古纹状体向脑桥及延髓的下行投射

以ＣＴＢ－ＨＲＰ为顺行示踪剂,经微电泳引入鸡一侧古纹状体,追踪其向脑桥和延髓的下行投射。结果观察到双侧的许多核团均有顺行标记终末出现,在及脑桥水平,出现于三叉神经运动核簇,面神经运动核簇、三叉神经感觉主核、上橄榄核、蓝斑核以及内、外侧桥核。在延髓内,出现于前庭核簇、耳蜗核簇、孤束核、下橄榄核、小细胞网状核、大细胞网状核、疑核、疑后核、舌下神经核、三叉神经脊核以及背索核。     

鸡古纹状体向脑桥及延髓的下行投射──CTB-HRP顺行追踪法研究

以ＣＴＢ－ＨＲＰ为顺行示踪剂，经微电泳引入鸡一侧古纹状体，追踪其向脑桥和延髓的下行投射。结果观察到双侧的许多核团均有顺行标记终末出现。在脑桥水平，出现于三叉神经运动核簇、面神经运动核簇、三叉神经感觉主核、上橄榄核、蓝斑核以及内、外侧桥核。在延髓内，出现于前庭核簇、耳蜗核簇、孤束核、下橄榄核、小细胞网状核、大细胞网状核、疑核、疑后核、舌下神经核、三叉神经脊核以及背索核（包括楔束外核）。     

鸡桥核向小脑的直接投射——HRP法研究

３０％ＨＲＰ水溶液注入鸡小脑的Ⅵ、Ⅶ、Ⅷ叶内，逆行追踪鸡桥核向小脑的投射。结果表明：鸡具有内、外两对桥核、各核均可投向小脑Ⅵ、Ⅶ、Ⅷ叶，说明鸡虽无明显的脑桥延髓界线，但确实存在脑桥部。与桥核同时出现的标记核团有蓝斑核、前庭上核、三叉神经脊束核极间亚核、网状结构等。     

鸡丘脑背外侧核纤维联系——HRP法研究

鸡丘脑背外侧核前部与上纹状体、新纹状体、外纹状体及圆核和视顶盖有纤维交互投射;丘脑背外侧核后部与上纹状体、新纹状体及圆核和视顶盖有纤维交互投射,且与丘脑其他核团有较多的联系。     

Heat inactivation of the neuraminidase and haemagglutinin estimated in the agglutination-separation reactions using red blood cells sensitized with Newcastle disease virus.

The agglutination-separation (AS) reactions estimate the effects of heat on the release of altered Newcastle disease virus (NDV) and HN glycoprotein spikes from red blood cells (RBC) sensitized with NDV (SRBC), the inactivations of the neuraminidase (NA), then the haemagglutinin (HA) in a direct assay. Heating SRBC for 1.5 min at 56 degrees C inactivated the NA by 50%; after 4.5 min no separation occurred indicating 100% inactivation of the NA. Heating a suspension of NDV for 78 min inactivated the NA 50% as assayed by cleavage of fetuin. Comparatively, the AS test was up to 52-fold (78 min/1.5 min = 52) more efficient in detecting NA inactivation than was the basic reference test where cleavage of fetuin was assayed. The HA was 50% inactivated after 18 min of heating and 100% inactivated after 36 min as no agglutination was seen. Free HA on SRBC was agglutinated by and thus was titrated with the sialic acid on NRBC. The large area of RBC increased the efficiency of the AS test when compared with tests using suspensions of NDV. At 51-60 degrees C all NA and HA inactivations were sequential, and invariably the NA was more heat labile than the HA. The release of altered NDV and HN spikes was inhibited with mild heat although the separation of SRBC and NRBC continued. Biological purifications showed that the heat stability of the HA and the lability of the NA were genetically stable.     

Effects of triamcinolone acetonide, sodium hyaluronate, amikacin sulfate, and mepivacaine hydrochloride, alone and in combination, on morphology and matrix composition of lipopolysaccharide-challenged and unchallenged equine articular cartilage explants

OBJECTIVE: To evaluate the effects of triamcinolone acetonide (TA), sodium hyaluronate (HA), amikacin sulfate (AS), and mepivacaine hydrochloride (MC) on articular cartilage morphology and matrix composition in lipopolysaccharide (LPS)-challenged and unchallenged equine articular cartilage explants. Sample POPULATION: 96 articular cartilage explants from 4 femoropatellar joints of 2 adult horses. PROCEDURES: Articular cartilage explants were challenged with LPS (100 ng/mL) or unchallenged for 48 hours, then treated with TA, HA, AS, and MC alone or in combination for 96 hours or left untreated. Cartilage extracts were analyzed for glycosaminoglycan (GAG) content by dimethyl-methylene blue assay (ng/mg of dry wt). Histomorphometric quantification of total lacunae, empty lacunae, and lacunae with pyknotic nuclei was recorded for superficial, middle, and deep cartilage zones. RESULTS: LPS induced a significant increase in pyknotic nuclei and empty lacunae. Treatment with TA or HA significantly decreased empty lacunae (TA and HA), compared with groups without TA or HA, and significantly decreased empty lacunae of LPS-challenged explants, compared with untreated explants. Treatment with AS or MC significantly increased empty lacunae in unchallenged explants, and these effects were attenuated by TA. Treatment with MC significantly increased empty lacunae and pyknotic nuclei and, in combination with LPS, could not be attenuated by TA. Content of GAG did not differ between unchallenged and LPS-challenged explants or among treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with TA or HA supported chondrocyte morphology in culture and protected chondrocytes from toxic effects exerted by LPS, AS, and MC.     

Agglutination-separation reactions of red blood cells sensitized with Newcastle disease virus: quantities, agglutination characteristics, and serology of altered virus and HN spikes released following neuraminidase reactivation

Red blood cells (RBC) become sensitized following the elution of strain 575 Newcastle disease virus (NDV). The neuraminidase (NA) in the haemagglutinin (HA)-sialic acid configuration is inactive. The HA on sensitized RBC agglutinates normal RBC. The sialic acid on normal RBC initiated reactivation of the NA-a newly described function. Then normal-sensitized RBC agglomerates separated at 37 degrees C in the irreversible agglutination-separation (AS) reactions. With separation the AS products. HN spikes (150-200 kDa) and altered NDV, which contain fewer HN spikes than intact allantoic NDV, were removed from the sensitized RBC and the NDV membrane. Extraction of HN spikes from the membrane required more sialic acid than the removal of AS products from RBC. Thus 2 reactions were delineated for the orderly removal. Amounts of each released AS product suggest the source of the HN spikes. AS reactions and ether treatment of NDV increased the HA titres up to 19.2 fold. HA-sialic acid configurations were estimated by the amounts of normal RBC agglutinated by sensitized RBC and also by agglutination with fetuin. Elution of B1 vaccine, HN spikes from ether-treated NDV as well as AS products separated on sephadex resulted in incompletely sensitized RBC; fewer configurations were titrated with normal RBC; all failed to respond to anti-NA antibody. In contrast, sensitized RBC or suspensions of 575 NDV, but not the B1 vaccine strain, responded to both anti-NA and anti-HA antibody. Sensitization, slow elution, and responding to anti-NA antibody, which was accompanied by fluorescent foci on sensitized RBC, required intact NDV and the infrequent HA-sialic acid configuration. The NA was inactive for the HA-sialic acid configuration but cleaved fetuin, indicating substrate specificity. An inactive NA would allow time for fusion and NDV penetration rather than elution by an active NA early in NDV infection.     

Changes in the distribution of labeled retinal ganglion cells after an implant of DiI into the optic nerve in the chick embryos

Changes in the distribution of retinal ganglion cells (RGCs) were studied using the retrograde labeling of DiI in chicks and chick embryos. The small retinal area filled with labeled RGCs was observed in the retinal fundus on E8. The labeled retinal area expanded radially toward the peripheral retina as the retina grew, and finally occupied a whole retina by P1. The temporal retina was labeled more rapidly than in the nasal retina. The observed-increasing rate of the labeled area was corrected with the growing rate of the retina. Consequently, the corrected-increasing rate of the labeled area was estimated to be about 390% between E8 and E11, and 20-50% after E11. This means that spreading speed of the maturated RGCs lowered until 1/10-1/20 after E11.     

Preparation of Atherosclerosis Model Using Guangxi Bama Mini-pig

The atherosclerosis (AS) model was prepared in Guangxi Bama mini-pig,and the atherosclerosis index (AI) was preliminarily identified in occurrence of AS to provide the basis for the preparation of related models.20 Guangxi Bama mini-pigs were randomly divided into control group and experimental group which were fed with high fat and cholesterol diet to prepare the AS model.Blood biochemical indexes were detected in the process of model preparation and the correlation between AI and the results of vascular slices was analyzed to initially draw up AI of AS.The results of vascular slices showed that the incidence rates of AS of Guangxi Bama mini-pigs in experimental group and control group were 20% and 0,respectively.The association analysis between the results of vascular slices and AI in the pathogenic Guangxi Bama mini-pigs preliminarily suggested that AI was above 3.8 and lasted for more than 3 months during the attack.     

广西巴马小型猪动脉粥样硬化模型的制作

本试验旨在制作广西巴马小型猪动脉粥样硬化模型,初步判断猪发生动脉粥样硬化时的指数值,以期为今后建立相关模型提供依据。试验选取广西巴马小型猪试验组和对照组各10头,通过饲喂高脂高胆固醇饲料建立动脉粥样硬化模型,检测建模过程中血液生化指标,将动脉粥样硬化指数与血管切片作关联分析,初步拟定发生动脉粥样硬化时的指数值。屠宰后血管切片结果显示,试验组有2头广西巴马小型猪发生动脉粥样硬化,对照组均正常。2头广西巴马小型猪的血管切片结果与动脉粥样硬化指数进行关联分析后,初步拟定的动脉粥样硬化指数值在3.8以上,并持续3个月以上。     

瑞士褐牛蜘蛛腿综合征致病位点检测方法的建立

牛蜘蛛腿综合征（Arachnomelia Syndrome,AS）是一种常染色体隐性遗传病,现主要见于德系西门塔尔牛和欧洲瑞士褐牛。患病犊牛先天骨骼系统严重畸形,出生即死亡或出生不久死亡。已证实瑞士褐牛蜘蛛腿综合征是由编码亚硫酸盐氧化酶的基因SUOX外显子4上一个单碱基插入突变c.363-364insG所致。本研究首次建立了瑞士褐牛蜘蛛腿综合征致病基因突变位点分子筛查的方法：荧光引物PCR产物电泳法和PCR产物直接测序法。使用荧光引物PCR产物电泳法对10个品种187头牛样本进行该突变位点的基因型分型,没有发现携带该致病基因的个体。荧光引物PCR产物电泳法判定基因型快速可靠,实验成本较低,适合大规模群体的检测。由于我国曾多次进口瑞士褐牛遗传物质,可能将该致病基因引入本地牛群体。因此对我国相关牛群开展AS隐性不利基因的筛查是必要的,尤其是针对种公牛进行筛查,可避免该致病基因在群体的传播造成重大经济损失。     

雏鸡投射向外侧膝状体腹侧核的视顶盖细胞形态研究

鸟类的一些视觉信息经由视顶盖Ⅰ层细胞（Ⅰ细胞）传递到外侧膝状体腹侧核（nucleus geniculatus lateralis ventralis,GLv）,用于色觉、瞳孔反射和视觉运动.使用羰花青荧光染料DiI逆向神经标记技术研究了这些投射向GLv核的视顶盖Ⅰ细胞的形态特征.根据标记细胞的胞体和树突野大小及树突分枝特点,雏鸡投射到GLv核的视顶盖Ⅰ细胞可分为“矛状树突Ⅰ细胞”和“叉状树突Ⅰ细胞”,前者具有小型纺锤状的细胞体和垂直伸延的1支尖端树突,后者具有较大三角形或多角形的细胞体和多支上行性树突.多数标记树突的末端部水平分布于视顶盖F层,和终止于此层的视神经终末部的形态相一致.     

猪流感病毒血凝素基因原核表达及其在间接ELISA中的初步应用

利用RT-PCR方法扩增猪流感病毒A/Swine/Henan/11/2005(H1N1)血凝素(HA)基因,克隆于pMD18-T载体进行测序。以pMD18-HA为模板、利用带酶切位点的引物再次扩增HA基因的开放阅读框(ORF),克隆到表达载体pET32a中,经双酶切、测序及PCR鉴定得到阳性重组表达质粒pET-HA,将质粒转化到表达宿主菌BL21(DE3)中,经终浓度为1 mmol/L的IPTG诱导,SDS-PAGE结果显示,HA蛋白获得了高效表达,经Western-blot检测证实表达产物具有良好的免疫学活性,在间接ELISA中的初步应用表明具有良好的抗原反应性。本研究为以重组HA蛋白为抗原建立H1亚型猪流感抗体的ELISA检测方法奠定了基础。