细胞化学

In the brain of adult rat, the positive immunohistochemical product of lSL-l (ISL-l-positive) was mainly located in the neuronal nucleus and found in discrete regions except to brain cortex, such as the Purkinje cell layer and the granular cell layer of cerebellum, the granular cell layer and the pyramidal cell layer of hippocampus, the mitral cell layer, the internal and external plexiform layer, the granular cell layer and the granular cell layer of olfactory bulb and so on, and several nuclei of the hypothalamus, midbrain and pons, such as claustrum, anterior olfactory nucleus, accumbens nucleus, caudate-ptamen, pallidum, substantia nigra, striatum, islands of Callaje, mammillary nucleus, anterior pretactal nucleus, habenular nucleus, amygdaloid nucleus, cuneate nucleus, rubral nucleus, gigantocellular reticular nucleus and so on.

在正常成年大鼠脑中，同源框基因islet-1表达产物（ISL-1）免疫组织化学阳性物质广泛分布于除大脑皮层外的神经细胞的细胞核内，ISL-1阳性神经元密集分布于小脑Purkinje细胞层和颗粒细胞层、海马的颗粒细胞层和锥体细胞层、嗅球的内丛层、外丛层、颗粒细胞层及僧帽细胞层等，另外在丘脑、中脑和桥脑的一些重要神经元核团均有分布，如，屏状核、前嗅核、伏核、尾壳核、苍白球、黑质、纹状体、Calleja岛、乳头体核、前顶盖前核、缰核、杏仁核、楔束核、红核网状巨细胞核等。

Antigene peptide nucleic acid specifically inhibits MYCN expression in human neuroblastoma cells leading to cell growth inhibition and apoptosis Roberto Tonelli,Stefania Purgato,Consuelo Camerin,Raffaele Fronza,Fabrizio Bologna,Simone Alboresi,Monica Franzoni,Roberto Corradini,Stefano Sforza,Andrea Faccini,Jason M. Shohet,Rosangela Marchelli,and Andrea Pession Department of Pediatrics, University of Bologna,Sant'Orsola-Malpighi Hospital, Bologna, Italy; Department of Organic and Industrial Chemistry, University of Parma, Parma,Italy; and 3Center for Cell and Gene Therapy, Texas Children's Cancer Center, Baylor College of Medicine, Houston, Texas Abstract We developed an antigene peptide nucleic acid for selective inhibition of MYCN transcription in neuroblastoma cells, targeted against a unique sequence in the antisense DNA strand of exon 2 of MYCN and linked at its NH2 terminus to a nuclear localization signal peptide.

抗原肽核酸抑制MYCN具体表达的神经母细胞瘤细胞，导致细胞生长的抑制和凋亡罗伯托利，斯特凡Purgato ，苏埃洛卡默琳，拉菲尔Fronza ，梅奥尼博洛尼亚，西蒙娜Alboresi ，莫妮卡弗兰索尼，罗伯托Corradini ，斯特凡诺斯福尔扎，安德烈Faccini ，杰森先生Shohet ， Rosangela Marchelli ，和安德烈Pession儿科，博洛尼亚大学，圣' Orsola -马尔皮基医院，博洛尼亚，意大利；有机部和工业大学化学帕尔马，帕尔马，意大利和3Center的细胞和基因治疗，德州儿童癌症中心， Baylor医学院，得克萨斯州休斯敦摘要我们开发了一种抗原肽核酸的选择性抑制神经母细胞瘤MYCN转录细胞，针对一个独特的序列反义DNA链的第2外显子，并与在MYCN 其氨基端为核定位信号肽。

In 8 cm grafted group , all survival dogs(n=5) were held upright with the affected hindlimb extended so that the body's weight was supported by the distal metatarsus and toes.

摘 要：［目的］研究神经长度能否影响化学去细胞神经的质量及化学去细胞异体神经修复犬神经缺损的适当长度范畴。

RESULTS: The positive staining cells mostly included ependymal cells of myelocoele, glial cells, motor neurons, and epithelial cells of spinal pia mater. The staining was mainly located on the cell membrane. Most importantly, it displayed certain directivity in the glial cells, mainly at the membrane side contacting with capillary endothelial cells.

结果： 免疫组织化学染色结果显示阳性着色细胞包括有胶质细胞、中央管室管膜上皮细胞、脊髓前角运动神经元、软脊膜上皮细胞，着色部位主要在细胞膜，其中在胶质细胞表现出一定的方向性，主要分布在与毛细血管内皮细胞接触的一侧。

Results: the positive cells mostly included ependymal cells of myelocoele, glial cells, anterior horn motor neurons, and epithelial cells of spinal pia mater and their expressions were mainly located on cell membrane. interestingly, aqp4 distribution in glial cells and ependymal cells showed certain directivity, but the former was mainly in the membrane side contacting with capillary endothelial cells and the latter mostly in the basolateral membrane of ependymal cells.

结果： 免疫组织化学染色和原位杂交结果显示胶质细胞、前角运动神经元、中央管室管膜细胞、软脊膜上皮细胞均表达阳性，aqp4主要分布于细胞膜，其中，胶质细胞和中央管室管膜细胞表达存在极性，前者主要分布在与毛细血管内皮细胞接触的一侧，后者主要分布在中央管室管膜细胞的基底侧膜。

The results of immuhistochemistry show: compared with normal retina, more 3-NT-positive cells and iNOS-positive cells appear in inner nucleus layer of diabetic retina; less eNOS-positive cells appear in inner nucleus layer and vascular endoderm of diabetic retina; less nNOS -positive cells appear in inner nucleus layer of diabetic retina; more ET-positive cells, ETRA- positive cells, ETRB- positive cells appear in inner nucleus layer of diabetic retina; moreα- synuclein- positive cells appear in ocular cone and rod layer of diabetic retina. Conclusions 1. RFDD-PCR is an efficient technique for research diseases genomics as a mass screening to complete gene expression with the identifying of candidate gene related to disease.

免疫组织化学结果显示：13-NT和NOS：与正常视网膜相比，8周糖尿病大鼠视网膜中，INL的3-NT和iNOS免疫阳性细胞明显增多，INL和血管内皮层的eNOS阳性细胞明显减少，INL的nNOS阳性细胞也明显减少；2ET及ETR：与正常视网膜相比，8周糖尿病大鼠视网膜中，ET、ETRA、ETRB免疫阳性细胞明显增多，增多的阳性细胞主要集中于INL，而在血管内皮层增多不明显；3α-synuclein：与正常视网膜相比，8周糖尿病大鼠视网膜中，α-synuclein免疫阳性细胞明显增多，增多的阳性细胞主要集中于视网膜视锥视杆层。

Using GSA-〓 lectin histochemistry, the first appearance of microglia occurred at E13, usually stained as scattered amoeboid cells. From then on, the number of microglia increased steadily, reached a peak at postnatal day 7 (P7), and then reduced gradually to adult level. Generally the morphological form of microglia undergoes transformation from early rounded or ameboid to adult resting ramified as they differentiate during development.

使用凝集素GSA-〓亲和组织化学染色，本实验最早观察到小胶质细胞在胚胎13天（E13），为散在分布的圆形或阿米巴样细胞，随后细胞数量逐渐增多；总体而言小胶质细胞的形态遵循着由圆形或阿米巴样逐渐转向纤细分枝状的演变规律，不同阶段细胞形态各有其特点，在不同区域也存在差异；小胶质细胞的数量在生后第七天（P7）达到高峰，其后开始逐步降低直至成年。

Inaddition,we extract total RNA from cultural U251 cell of humanglioma,make coding gene extron amplification of TfR through ReverseTranscription-Polymerase Chain Reaction,construct expressionplasmid of pCDNA3.1-TfR after digestion of enzyme andconjunction,do transfection to U251 cell by liposome afteridentification,cause excess expression,detect the efficiency of transientexpression of TfR protein by pEGFPN1-TfR, sieve positive cell ofpCDNA3.1-TfR through G418, choose positive cell clone, detect theexpression of TfR in anteroposterior period through Western blot,FCM,cell immunochemistry and mRNA through RT-PCR, identify the effect of transfection.

另外，培养人脑胶质瘤U251细胞，进行细胞总RNA抽提，运用RT-PCR方法进行TfR基因的完整编码区外显子的扩增，经过酶切、连接后构建pCDNA3.1-TfR的表达质粒，鉴定正确后按脂质体转染方法转染人胶质瘤U251细胞，使其过量表达，通过pEGFPN1-TfR检测其瞬时转染的效率；用G418对稳定表达pCDNA3.1-TfR的细胞进行筛选，挑选阳性细胞克隆，运用Western blot、流式细胞检测技术和细胞免疫化学以及mRNA半定量的方法检测TfR在转染前后的表达水平，鉴定转染的效果。

We introduced improved primary mixed glial culture and different-attachment method to isolate and purify the OPCs, the cells were proliferated in serum-free medium, flow cytometry and immunohischemistry methods were employed to estimate the purity of cultured OPCs. Their abilities of differentiation and expression of trophic factors were identified by RT-PCR and immunostaining. Several methods including TUNEL and MTT were adopted to estimate the protective effects of conditioned culture medium from oligodendrocyte lineage cells on the primary cultured cerebellar granule neurons. Intravitreal transplant of OPCs, combined with retrograde fluorescent labeling the superior colliculus and intraorbital optic nerve transection, were used to investigate the protective effects of OPCs on the axotomized RGCs in vivo. Intravitreal transplantof OPCs or NSCs on the newborn rats, and retinal transplant of OPCs on the young rats were performed, to observe the myelin formation in the retina at different stages after cellular transplantation. Optic nerve transection was carried out on some rats with myelinated retinae, to study the influence of myelination on the injuried RGCs.

为此，本研究采用改良的胶质细胞混合培养与差速贴壁方法获得大鼠OPCs，使用无血清培养基进行扩增、培养，用免疫组织化学和流式细胞技术对培养细胞的纯度进行鉴定，对少突胶质系细胞表达部分营养因子的情况进行检测；采用TUNEL、MTT等方法对少突胶质系细胞条件培养基对原代培养小脑颗粒神经元的保护作用进行检测；将OPCs移植入成年SD大鼠玻璃体内，利用上丘逆行荧光标记技术，观察眼内移植的OPCs对眶内视神经切断时的视网膜神经节细胞的保护作用及其持续时间；将OPCs或NSCs移植入新生和幼年SD大鼠玻璃体或视网膜内，观察不同时期视网膜内髓鞘形成与分布特点，分析髓鞘的超微结构，并观察眼内髓鞘形成对损伤神经节细胞的保护作用。

Before the two-hour interaction of male-male individuals , we found there were obviously fewer and lighter stained ERp immunoreactive neurons (ER|3-IRs) in both main olfactory bulb system project regions and vomeronasal system project regions in the mandarin voles than in the reed voles, but no significant difference in numbers of AR immunoreactive neurons between the two species through the examining of immunoreactive neurons, moreover, there were significantly fewer AR-IRs than ERp-IRs in two species of vole before the two-hour interactions.

免疫组织化学检测，没有社会交往时棕色田鼠主嗅球系统投射区和犁鼻系统投射区ERβ免疫阳性细胞明显少于沼泽田鼠，且显色淡，AR免疫阳性细胞在两种鼠间差异不大，且都明显少于各自的ERβ-IRs.2h交往后，棕色田鼠主嗅球投射区和犁鼻系统投射区的ERβ-IRs细胞数明显少于交往前，AR-IRs细胞数明显多于交往前；沼泽田鼠交往前与交往后ERβ-IRs和AR-IRs细胞数均无显著差异，且显著多于交往后棕色田鼠ERβ-IRs细胞数，显著少于交往后棕色田鼠AR-IRs细胞数。

I can not make it blossom and suits me

我不能让树为我开花

When temperatures are above approximately 80 °C discolouration of the raceways or rolling elements is a frequent feature.

当温度高于 80 °C 左右时，滚道或滚动元件褪色是很常见的特征。