极北鲵肠内5-羟色胺、胃泌素和生长抑素内分泌细胞胚后发育

目的 观察极北鲵胚后发育期间肠内5-羟色胺(5-HT)、胃泌素(gastrin,GAS)和生长抑素(somatostatin,SS)3种内分泌细胞的分布、形态和数量变化。方法 应用亲和素-生物素复合物(avidin-biotin complex,ABC)法免疫组织化学技术,对38-46期健康极北鲵肠内5-羟色胺(5-HT)、胃泌素(GAS)和生长抑素(SS)细胞进行染色,并统计分析不同时期极北鲵肠内3种内分泌细胞的密度。结果 5-HT免疫反应阳性细胞和GAS免疫反应阳性细胞最先在41期检出,而SS免疫反应阳性细胞在42期才检测到。5-HT细胞在41期最少,46期最多,呈年龄相关的递增趋势;GAS细胞在43期最少,44期最多,呈先减后增变化趋势;SS细胞在43期密度高于其它各时期。5-HT细胞最先分布在上皮基部,GAS细胞和SS细胞最先在肠上皮之间检测到。细胞形态多样,有椭圆形、三角形和锥体形等,但以锥体形为主。结论 极北鲵胚后不同发育时期肠内3种内分泌细胞的形态与其个体生理活动的改变相适应,且在数量变化上有其种属特异性。     

极北鲵消化道6种内分泌细胞的免疫组织化学研究

应用卵白素-生物素-过氧化物酶复合物(ABC)免疫组织化学法,对极北鲵(Salamandrella keyserlingii)消化道5-羟色胺(5-HT)、胃泌素(Gas)、生长抑素(SS)、胰高血糖素(Glu)、人胰多肽(PP)和P-物质(SP)6种内分泌细胞的分布进行了研究。文中采用Duncan多重比较的方法,对消化道内分泌细胞的分布密度进行统计学分析。结果显示,5-羟色胺细胞在消化道各部位均有分布,胃体(2.80±0.70)和十二指肠(2.60±0.75)分布密度最高,幽门最低(0.85±0.67);胃泌素细胞仅分布于小肠,十二指肠(1.85±0.75)密度最高;生长抑素细胞分布于食管、胃体、幽门、十二指肠和回肠各部位,且幽门(2.25±0.64)分布密度最高;胰高血糖素细胞只见于胃贲门部和胃体,人胰多肽细胞和P-物质细胞在整个消化道均未检测到。各种内分泌细胞的形态多样,呈圆形、椭圆形、锥体形和梭形等。极北鲵消化道内分泌细胞的这种密度分布和形态特点与其他两栖类动物及黑龙江产地的极北鲵相比,既有共性又有差异,其原因可能与物种、食性和栖息地环境等因素有关。     

极北鲵消化道嗜银细胞的胚后发生

为了探讨极北鲵(Salamandrella keyserlingii)消化道嗜银细胞胚后发生的形态学特征及分布规律,采用Grimelius银染法,对30～46期的极北鲵幼体进行了研究。结果显示,嗜银细胞在食管、胃、小肠各部分的发生时间不同。胃中的嗜银细胞在第41期出现,小肠中的在第42期出现,食管中的在第43期出现。嗜银细胞形态多样,有圆形、椭圆形、梭形和锥体形,大多分布在消化道黏膜上皮之间。根据嗜银细胞的形态判定其可能具有内分泌和外分泌两重功能。极北鲵胚后发育消化道各部位嗜银细胞出现顺序的不同可能与其消化生理活动及个体生理活动的变化相适应。     

黑龙江林蛙胃肠道5-羟色胺细胞的胚后发生

采用亲和素-生物素复合物(ABC)免疫组织化学法,对黑龙江林蛙(Rana amurensis)胚后发育各时期胃肠道中5-羟色胺(5-HT)细胞的发生过程进行了研究。结果显示,黑龙江林蛙胃中的5-羟色胺阳性细胞最早在第38期被检测到,小肠和大肠最早出现在第41期。从41期开始,5-羟色胺细胞分布于胃、小肠和大肠中,在同一发育时期中,胃部分布密度最大。同一部位在不同发育时期5-羟色胺细胞的分布密度不同,其中胃内5-羟色胺细胞在38期和43期分布最多,分布密度分别为1.8±0.62和1.8±0.89,45期分布数量最少,分布密度为1.1±0.31;小肠中5-羟色胺细胞在41期和43期分布最多,分布密度均为1.2±0.41,在44期和45期分布最少,分布密度分别为0.7±0.49和0.7±0.47;大肠中5-羟色胺细胞在第42期分布最多,分布密度为1.2±0.41,在44期分布最少,分布密度为0.7±0.47。5-羟色胺细胞最早分布在上皮细胞之间,随着胃肠道发育的日渐完善,在上皮基部或腺泡上皮之间也有分布。细胞形态变化不明显,有圆形、椭圆形、锥体形、长梭形和三角形。推测5-羟色胺阳性细胞的出现时间、形态以及分布型是与黑龙江林蛙的个体生长发育需要、胃肠道各段的功能以及食性转变相适应的。     

乌龟胃肠胰系统内分泌细胞的免疫组织化学研究

应用过氧化物酶标记的链霉卵白素(Streptavidin peroxidase,简称S-P法)免疫组织化学技术,使用六种特异性胃肠激素抗血清对乌龟胃肠胰系统内分泌细胞的种类、定位、分布密度及形态进行了研究。在乌龟胰腺中检测出5-羟色胺、生长抑素、胰高糖素和胰多肽等4种内分泌细胞,生长抑素、胰高糖素细胞多成簇大量分布于胰岛中;5-羟色胺、胰多肽细胞多散在少量分布于胰腺腺泡之间。在乌龟消化道中共检测出5-羟色胺、生长抑素、胃泌素、胰高糖素和P物质等5种内分泌细胞:5-羟色胺细胞在消化道各段均有分布,以十二指肠处分布密度最高(30.7±4.2),空肠其次,回肠、直肠处最低(12.0±1.0/11.2±3.0);生长抑素细胞仅分布于食道和胃中各段;胃泌素细胞分布于胃幽门部和十二指肠处;胰高糖素细胞分布于胃体至空肠段,以胃幽门部分布密度较高(11.3±1.1);P物质细胞仅布于胃幽门部;消化道各段均未检出胰多肽细胞。与其他爬行动物比较,乌龟胃肠胰系统内分泌细胞的分布既存在着一定共同点,又显示了较大的种间差异。       

白条草蜥消化道内分泌细胞的免疫组织化学

应用6种胃肠激素抗血清和免疫组织化学ABC法(avidin-biotin complex method),对白条草蜥(Takydromus wolteri)消化道内分泌细胞进行了免疫组织化学定位研究和形态学观察。结果表明,5-羟色胺细胞较其他5种内分泌细胞的分布更为广泛,整个消化道中(即从食管到直肠)均有分布,其分布密度高峰位于幽门。食管、回肠和直肠未检测到生长抑素细胞,生长抑素细胞在幽门部分布密度最高,总体来说生长抑素细胞的分布在胃部较高而在小肠部较低。胃泌素细胞和胰多肽细胞分布在小肠,均在十二指肠分布密度最高。胰高血糖素细胞在胃幽门部分布密度最高,十二指肠、空肠次之,回肠分布密度最低。P-物质细胞仅分布于幽门部。6种内分泌细胞以圆形和锥体形为主,它们广泛分布于消化道黏膜之间、腺泡上皮细胞之间及上皮细胞基部。内分泌细胞的密度分布与其食性、食物组成和生活环境有关,它们的形态与其内、外分泌功能是相适应的。     

蝘蜓消化管内分泌细胞的免疫组织化学定位

目的阐明爬行动物蝘蜓消化管各段内分泌细胞的类型、局部分布、分布密度和形态学特征。方法应用免疫组织化学技术中链霉卵白素-过氧化物酶(S-P)法。结果在蜓消化管内鉴别出5种内分泌细胞,即:5-羟色胺(5-hydroxytryptamine,5-HT)、生长抑素(somatostatin,SS)、胃泌素(gastrin,GAS)、高血糖素(glucagon,GLU)、P-物质(substance P,SP)免疫活性(i mmnoreactive,IR)细胞。5-羟色胺免疫活性细胞是消化管中最主要的内分泌细胞类型,以不同密度分布于消化管各段,其中胃幽门部位分布密度最高。生长抑素免疫活性细胞在消化管内仅局限分布于胃部。胃泌素免疫活性细胞仅见于幽门和十二指肠部位。高血糖素免疫活性细胞仅分布于回肠和直肠。P-物质免疫活性细胞仅出现在直肠部位。蝘蜓消化管内分泌细胞形态多样:圆形、椭圆形、纺锤形、梭形、楔形、锥形以及不规则形。胃部多数内分泌细胞分布于胃腺中,食管和肠管中多数内分泌细胞则分布于上皮细胞间。结论蜓和其它爬行动物胃肠内分泌细胞的分布存在一定共同特征,然而也存在着一些特有的差异。     

红腹锦鸡胃肠道内分泌细胞的免疫组织化学定位

应用ABC免疫组织化学方法,对红腹锦鸡(Chrysolophus pictus)胃肠道5-羟色胺(5-hydroxtryptamine,5-HT)、胃泌素(gastfin,GAS)、生长抑素(somatostatin,ss)3种内分泌细胞的分布密度和形态进行了观察.结果显示,5-HT细胞在空肠和直肠分布密度最高,回肠和盲肠次之,十二指肠较少,腺胃和肌胃最少;GAS细胞在十二指肠和直肠分布密度最高,其次是空肠和盲肠,腺胃部最低,肌胃则呈免疫阴性;SS细胞数量较少,在直肠、盲肠处分布密度相对高,其次是十二指肠和空肠,腺胃部最低,肌胃则呈免疫阴性.3种内分泌细胞的形态多样,有圆形、椭圆形、锥体形、杆状和不规则形,其中以圆形、椭圆形为主.细胞分布于固有膜、黏膜上皮细胞基部、黏膜上皮细胞之间、腺泡上皮细胞基部或腺泡上皮细胞之间.红腹锦鸡胃肠道内分泌细胞的形态与其内、外分泌功能是相适应的.     

蝘蜒消化管内分泌细胞的免疫组织化学定位

目的阐明爬行动物蝘蜒消化管各段内分泌细胞的类型、局部分布、分布密度和形态学特征。方法应用免疫组织化学技术中链霉卵白素一过氧化物酶（S-P）法。结果在蝘蜒消化管内鉴别出5种内分泌细胞,即：5—羟色胺（5-hydroxytryptamine,5-HT）、生长抑素（somatostatin,SS）、胃泌素（gastrin,GAS）、高血糖素（glucagon,GLU）、P-物质（substance P,SP）免疫活性（immnoreactive,IR）细胞。5-羟色胺免疫活性细胞是消化管中最主要的内分泌细胞类型,以不同密度分布于消化管各段,其中胃幽门部位分布密度最高。生长抑素免疫活性细胞在消化管内仅局限分布于胃部。胃泌素免疫活性细胞仅见于幽门和十二指肠部位。高血糖素免疫活性细胞仅分布于回肠和直肠。P-物质免疫活性细胞仅出现在直肠部位。蝘蜒消化管内分泌细胞形态多样：圆形、椭圆形、纺锤形、梭形、楔形、锥形以及不规则形。胃部多数内分泌细胞分布于胃腺中,食管和肠管中多数内分泌细胞则分布于上皮细胞间。结论蝘蜒和其它爬行动物胃肠内分泌细胞的分布存在一定共同特征,然而也存在着一些特有的差异。     

变色树蜥消化道5-羟色胺细胞的免疫组织化学定位

用免疫组织化学ABC法,应用5-羟色胺(5-HT)特异性抗血清,对变色树蜥(Calotes versicolor)消化道内含有5-HT的内分泌细胞进行了免疫组织化学的定位研究和形态学观察。结果显示,5-HT细胞在变色树蜥消化道的各个部位均有分布,分布密度近似呈“M”形,其中以空肠分布密度最高,胃体次之,食管最低。5-HT细胞的形态多样,呈圆形、椭圆形、锥体形、长锥体形等,其中胃、幽门和直肠以圆形和椭圆形为主,小肠则以长锥体形为主,其细长突起指向肠腔或固有膜;细胞分布于上皮基部、上皮细胞之间、腺泡上皮细胞之间或固有膜内;多数细胞以内分泌功能为主,少数细胞具有外分泌功能。比较分析表明,5-HT细胞分布型可能与动物的食性有关。     

胚胎干细胞起源的探讨

目前胚胎干细胞(ESCs)建系的取材来源包括桑椹胚的卵裂球、囊胚的内细胞团(ICM)、上胚层细胞和原始生殖细胞(PGCs),甚至从新生鼠睾丸细胞也分离得到类ES样细胞系。这就提出了一个问题,什么是ESCs最接近的体内细胞来源。传统观念常常把ESCs等同于ICM细胞,也有学者认为ESCs更象上胚层细胞,而在已知的分子标记基因方面,ESCs所具有的特征更接近体内早期生殖细胞。不清楚ESCs最接近的体内细胞来源,可能是制约许多品系小鼠和大多哺乳类动物建系成功率提高的原因之一。ESCs系与EG细胞系的分离条件不同表明,加强对ESCs多能性维持基因调控研究具有重要意义。本文从ESCs的经典概念及其发展,早期胚胎细胞和生殖细胞发育规律,早期胚胎细胞、早期生殖细胞和ESCs的关系等方面进行综合分析,认为ESCs可能有多种接近的体内细胞来源。进一步应通过对ESCs建系不同的取材细胞和不同品系的ESCs间进行比较研究,以便弄清ESCs的来源和转化机制,为提高不同物种ESCs建系效率提供理论支持。     

Fine needle aspiration (FNA) cytology of pilomatrixoma

FNA smears from five histologically confirmed cases of pilomatrixoma were reviewed to delineate the cytological features helpful in diagnosis. A combination of basaloid cells, ghost cells and foreign body giant cells appeared to be necessary in FNA smear for a confident cytodiagnosis of pilomatrixoma. Presence of naked nuclei, nucleated squamous cells and calcification were additional features in favour of the diagnosis. Another 10 cases with initial cytodiagnosis of pilomatrixoma or benign skin appendage tumour were reviewed. Using the above criteria, diagnosis of pilomatrixoma was easy in five cases. One case was problematical due to presence of atypical squamous cells. Initially the cytological features were most commonly confused with epidermal inclusion cyst, giant cell lesion or a squamous cell carcinoma. The main reasons for erroneous diagnosis were lack of awareness of cytological features, predominance of one component over the others, and non‐representative FNA smears. Atypia in nucleated squamous cells, and misinterpretation of basaloid cells as malignant can lead to diagnostic dilemma. Adequate clinical data are also necessary.     

A comparison of the tube forming potentials of early and late endothelial progenitor cells

The identification of circulating endothelial progenitor cells (EPCs) has revolutionized approaches to cell-based therapy for injured and ischemic tissues. However, the mechanisms by which EPCs promote the formation of new vessels remain unclear. In this study, we obtained early EPCs from human peripheral blood and late EPCs from umbilical cord blood. Human umbilical vascular endothelial cells (HUVECs) were also used. Cells were evaluated for their tube-forming potential using our novel in vitro assay system. Cells were seeded linearly along a 60 μm wide path generated by photolithographic methods. After cells had established a linear pattern on the substrate, they were transferred onto Matrigel. Late EPCs formed tubular structures similar to those of HUVECs, whereas early EPCs randomly migrated and failed to form tubular structures. Moreover, late EPCs participate in tubule formation with HUVECs. Interestingly, late EPCs in Matrigel migrated toward pre-existing tubular structures constructed by HUVECs, after which they were incorporated into the tubules. In contrast, early EPCs promote sprouting of HUVECs from tubular structures. The phenomena were also observed in the in vivo model. These observations suggest that early EPCs cause the disorganization of pre-existing vessels, whereas late EPCs constitute and orchestrate vascular tube formation.     

树突状细胞与CIK细胞共培养诱生的DCCIK细胞群对肿瘤的杀伤作用

由树突状细胞(DC)与细胞因子诱导的同源杀伤细胞(CIK)的共培养诱生的细胞群(DCCIK)对肿瘤细胞的细胞毒活性的研究。DCCIK细胞体外杀伤肿瘤靶细胞A549(MTT法),效靶比为10∶1、5∶1时杀伤率分别为61%、52%。DCCIK细胞诱导培养3周后,效靶比为10∶1、5∶1时杀伤率分别为64%和56%。数据亦表明DCCIK细胞对靶细胞的杀伤优于CIK细胞。动物体内实验分荷瘤A549、BEL7404和A375三组,每组分(A)DCCIK 化疗、(B)单用化疗。治疗20天、35天后测量各组肿瘤消失率。结果显示:DCCIK 化疗的抑瘤效果明显好于单纯化疗。提示DCCIK细胞有临床应用前景。     

Resistance and augmentation of innate immunity in mice exposed to starvation

Mice were exposed to starvation for 3 days. Body temperature and various parameters were examined. By starvation, body temperature, blood glucose and ACTH decreased, especially on days 2 and 3. The level of corticosterone increased at this time. On the other hand, the number of lymphocytes yielded by the liver, spleen and thymus decreased from day 1 to 3. The change of the distribution of lymphocyte subsets was unique because NK, NKT and extrathymic T cells were stress-resistant in the liver. Conventional T and B cells were stress-sensitive. Reflecting the increased proportion of NK and NKT cells, NK and NKT activities were augmented. The increased proportion of NKT cells produced both IFNγ and IL-4 (Th0-type profile). The proportion and some functions of granulocytes and macrophages increased on Day 1 after starvation. These results suggest that starvation has a potential to increase the functions of unconventional lymphocytes and myeloid cells.     

Ultrastructure of the surface of the iris in the rat eye

Summary The surface structure of the iris in the rat eye was studied by light and electron microscopy.The anterior surface of the rat iris is covered with a discontinuous layer of large, polygonal endothelial cells with microvilli on their surface. Crypts and holes between adjacent endothelial cells extend into the stroma and form there a complicated network of interconnected spaces occupying about one half or more of the volume of the pupillary part of the stroma. The crypts are occasionally partly covered with endothelial cells. The posterior surface is covered with a continuous layer of polyhedronal epithelial cells. These are covered with many folds and processes, partly masked by an amorphous coat. The sphincter pupillae and dilatator muscles are possible to recognize on the scanning electron micrographs as well as blood vessels and nerve fibers in the iris stroma.The endothelial cells show many structural similarities with the endothelial cells on the cornea, probably reflecting their common origin. The results obtained, especially those from the scanning electron microscopic studies, are discussed and interpreted in relation to previous studies. The advantages in using different light and electron microscopic techniques are stressed.Supported by grants from Magnus Bergwall's Stiftelse and the Swedish Medical Research Council (B71-12X-2543-03).     

猪多能干细胞的研究进展

多能干细胞,如胚胎干细胞（embryonic stem cells,ESCs）、诱导多能干细胞（induced pluripotent stem cells,iPSCs）和成体干细胞（adultstemcells,ASCs）,是一类具有巨大潜能的独特细胞。猪作为试验材料,在遗传、代谢、生理生化及基因序列等方面较小鼠更接近于人类,正逐渐成为人类异种移植和再生医学研究的理想生物学模型。然而,目前对猪多能干细胞种类、来源、特征及机制的有限认识直接阻碍了其相关应用。该文将分别对猪ASCs的研究现状、猪类ESCs的分离培养、猪iPSCs的研究进展、多能干细胞间的联系和展望进行论述,以期为从事该领域研究的科研人员提供参考。     

Enhancement of BDNF production by co-cultivation of human neuroblastoma and fibroblast cells

It has been proved that co-cultivation of human neuroblastoma cells and human fibrolast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76×10⁶ viable cells/mL from 9×10⁵ viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5×10⁶ viable cells/mL, which was much higher than that from fed-batch cultivation. The nerve cell growth was greatly enhanced in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from, human fibroblast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.