草鱼垂体STH细胞组织化学和超微结构研究

本文分别对草鱼性成熟前、后垂体ＳＴＨ细胞进行了组织化学和超微结构研究。垂体ＳＴＨ细胞多位于中腺垂体中部和背部,为嗜酸性细胞,用ＰＭＢ（ＰＡＳ－ＭＢ）和ＡＰＧ（ＡＢ－ＰＡＳ－ＯＧ）两种组织化学方法染色,对橙黄Ｇ阳性,对ＰＡＳ、ＡＢ阴性;电镜下电子密度较高,内质网绕核呈环形,分泌颗粒多而小。     

基因修饰的小鼠肥大细胞瘤细胞的酶细胞化学变化

应用酶细胞化学方法对基因转导前后的Ｐ８１５小鼠肥大细胞瘤细胞进行酸性磷酸酶、非特异性酯酶及多糖的定位。研究结果显示,空载体转导的Ｐ８１５／ｎｅｏ细胞与其亲本细胞相比无显著改变,而ＩＬ２基因修饰的Ｐ８１５细胞的酸性磷酸酶及多糖反应明显增强,可能提示细胞内某些细胞器的形成以及细胞内多糖类合成功能的增强。     

石刁柏非胚性细胞与体细胞胚胚体细胞的超微结构研究

对石刁柏（Ａｓｐａｒａｇｕｓｏｆｆｉｃｉｎａｌｉｓ）体细胞胚发生过程中细胞的超微结构进行了观察,非胚性细胞内液泡大,大量的自体吞噬泡出现,胚性细胞内细胞核大,核移中,核仁结构明显,线粒体、质体、核糖体、高尔基体、内质网等细胞器增多,淀粉、脂滴积累,有较活跃的自体吞噬现象,梨形细胞内质体向叶绿体转变。     

实验性脾虚证大鼠内分泌腺形态学与细胞化学研究

成年Ｗｉｓｔａｒ大鼠２０只,分对照组和实验组。实验组用大黄煎剂灌服４２天致实验性脾虚证。对两组睾丸,肾上腺和甲状腺进行常规形态学与细胞化学观察。形态学显示：睾丸,肾上腺和甲状腺的组织和细胞均有不同程度的损伤。细胞化学显示：各内分泌腺均表现酸性磷酸酶（ＡＣＰ）减弱,个别内分泌腺细胞显示琥珀酸脱氢酶（ＳＤＨ）,三磷酸腺苷酶（ＡＴＰａｓｅ）和ＰＡＳ反应的变化。结果证明脾虚证不仅消化系统功能低下,而且内分泌功能也受到了影响     

不同年龄段前列腺外周带基质细胞超微结构差异

目的：研究不同年龄段前列腺外周带基质细胞超微结构并探讨其意义。方法：透射电镜观察3例年轻组（23-、26-、32-岁）和3例老年组（56-、64-、71-岁）正常前列腺外周带和3例前列腺癌组织的超微结构。Masson＇s Trichrome染色法观察前列腺外周带基质胶原的分布情况。结果：年轻组、老年组前列腺外周带间质均以平滑肌细胞分布为主,间杂成纤维细胞,但老年组平滑肌细胞和成纤维细胞体积较大、形态不规则。与正常年轻组相比,老年组前列腺外周带成纤维细胞存在明显老化和肌化改变,部分细胞内与蛋白合成相关的细胞器粗面内质网发达,呈现肌成纤维细胞特征。与年轻组和老年组外周带相比,前列腺癌基质中功能更加活跃的肌成纤维细胞和胶原蛋白数量增加明显。Masson＇s Trichrome染色支持超微观察到的基质胶原分布差异：年轻组〈老年组〈前列腺癌组。结论：不同年龄段前列腺外周带基质细胞超微结构间存在差异,老年组前列腺外周带和前列腺癌基质组织中具有活性的肌成纤维细胞的增加可能是造成老年前列腺癌高发和恶性进展的重要病理基础之一。     

不同年龄段前列腺外周带基质细胞超微结构差异

目的:研究不同年龄段前列腺外周带基质细胞超微结构并探讨其意义。方法:透射电镜观察3例年轻组(23-、26-、32-岁)和3例老年组(56-、64-、71-岁)正常前列腺外周带和3例前列腺癌组织的超微结构。Masson’s Trichrome染色法观察前列腺外周带基质胶原的分布情况。结果:年轻组、老年组前列腺外周带间质均以平滑肌细胞分布为主,间杂成纤维细胞,但老年组平滑肌细胞和成纤维细胞体积较大、形态不规则。与正常年轻组相比,老年组前列腺外周带成纤维细胞存在明显老化和肌化改变,部分细胞内与蛋白合成相关的细胞器粗面内质网发达,呈现肌成纤维细胞特征。与年轻组和老年组外周带相比,前列腺癌基质中功能更加活跃的肌成纤维细胞和胶原蛋白数量增加明显。Masson’s Trichrome染色支持超微观察到的基质胶原分布差异:年轻组<老年组<前列腺癌组。结论:不同年龄段前列腺外周带基质细胞超微结构间存在差异,老年组前列腺外周带和前列腺癌基质组织中具有活性的肌成纤维细胞的增加可能是造成老年前列腺癌高发和恶性进展的重要病理基础之一。     

双哌达莫促肺泡巨噬细胞抗病毒作用

观察双哌达莫 （ＤＰＭ） 对呼吸道合胞病毒 （ＲＳＶ） 肺炎小鼠肺泡巨噬细胞 （Ａｍ ） 的作用。采用组织化学和电镜等方法, 对肺炎组与肺炎＋ ＤＰＭ 处理组进行了对比观察。结果显示肺炎组Ａｍ 超微结构发育不良, 吞噬与分泌能力弱; ＤＰＭ 处理组比肺炎组Ａｍ 数量轻度增加, 细胞内溶酶体、滑面内质网与线粒体丰富,吞噬消化、分泌能力增强,病毒滴度与ＲＮＡ阳性细胞明显减少（Ｐ＜ ００１ 与Ｐ＜ ００５）。ＤＰＭ 提高Ａｍ非特异性免疫功能     

草鱼肾脏和脾脏血细胞发育过程的观察

鱼类肾脏、脾脏是机体造血的主要器官。印迹涂片显示:草鱼肾脏和脾脏内血细胞的发育过程大致经历了三个阶段,即原始阶段、幼稚阶段、成熟阶段。本文着重描述了各阶段细胞的形态特征井对草鱼血细胞的发育及命名等问题作了初步的探讨。       

天然杀伤细胞在受照射小鼠中的造血调控作用

研究了天然杀伤（ＮＫ）细胞对受致死剂量γ线照射的同系小鼠的造血调控作用。ＡＭＳ／５小鼠经９Ｇｙγ线全身照射后立即经尾静脉注射ＮＫ细胞（５×１０５）,可明显提高受照小鼠３０ｄ活存率,照后８ｄ小鼠骨髓中ＣＦＵ－ＧＭ数量明显高于对照和脾细胞注射组,照射后３０ｄ,ＮＫ细胞注射组活存小鼠的骨髓有核细胞数和ＣＦＵ－ＧＭ数已恢复到正常的７６％─９６％。病理组织学观察显示,输注ＮＫ细胞可使小鼠骨髓、脾脏的组织损伤程度减轻,造血功能增强,表现为造血灶数增多,造血细胞功能活跃,核分裂相增多,且涉及红系、粒系、巨核细胞系造血。ＮＫ细胞可能通过直接与造血干细胞相互作用或改善造血微环境等促进“内源性”造血功能,从而发挥对造血的正调控作用。提示ＮＫ细胞在小鼠造血功能的平衡维持中起重要作用。     

RCS大鼠和Wistar大鼠视网膜酸性磷酸酶活性的动态观察

本实验观察了不同年龄组ＲＣＳ大鼠和Ｗｉｓｔａｒ大鼠视网膜中酸性磷酸酶的动态变化及其与ＲＰＥ细胞消化功能的关系。运用偶氮偶联法显示１２ｄ、２１ｄ、２ｍ的ＲＣＳ大鼠和７ｄ、２ｍ的Ｗｉｓｔａｒ大鼠视网膜中的酸性磷酸酶;通过图像分析仪测定ＲＰＥ细胞层和光感受器外节部分的酸性磷酸酶含量,并进行统计学分析。结果：酸性磷酸酶阳性反应呈暗红色,主要位于ＲＰＥ细胞层,视网膜外核层、内核层,节细胞层亦有少量阳性反应颗粒。２ｍ的ＲＣＳ大鼠视细胞内、外节的酸性磷酸酶含量则明显高于其它组（Ｐ＜０．０１）,其余结构的酸性酶各组间无显著性差异（Ｐ＞０．０５）。结论：ＲＣＳ大鼠和Ｗｉｓｔａｒ大鼠的视网膜色素上皮细胞可能具有相同的消化功能。     

Haematopoietic stem cells in spleen have distinct differentiative potential for antigen presenting cells

Dendritic cells (DC) are known to develop from macrophage dendritic progenitors (MDP) in bone marrow (BM), which give rise to conventional (c)DC and monocytes, both dominant antigen presenting cell (APC) subsets in spleen. This laboratory has however defined a distinct dendritic‐like cell subset in spleen (L‐DC), which can also be derived in long‐term cultures of spleen. In line with the restricted in vitro development of only L‐DC in these stromal cultures, we questioned whether self‐renewing HSC or progenitors exist in spleen with restricted differentiative capacity for only L‐DC. Neonatal spleen and BM were compared for their ability to reconstitute mice and to give rise to L‐DC, as well as other splenic APC. Neonatal spleen cells were transplanted into allotype‐distinct lethally irradiated hosts along with host‐type competitor BM cells, and assayed over 8 to 51 weeks for haematopoietic reconstitution of L‐DC and cDC subsets, along with other lymphoid and myeloid cells. In this study, neonatal spleen showed multilineage haematopoietic reconstitution in mouse chimeras, rather than specific or restricted ability to differentiate into L‐DC. However, the representation of individual APC subsets was found to be unequal in chimeras partially reconstituted with donor cells, such that more donor‐derived progeny were seen for L‐DC than for myeloid and cDC subsets. The ability of HSC in spleen to develop into L‐DC was indicated by a strong bias in the subset size of these cells over other splenic APC subsets. This type of evidence supports a model whereby spleen represents an important site for haematopoiesis of this distinct DC subset. The conditions under which haematopoiesis of L‐DC occurs in spleen, or the progenitors involved, will require further investigation.     

The peripheral chimerism of bone marrow–derived stem cells after transplantation: regeneration of gastrointestinal tissues in lethally irradiated mice

Bone marrow–derived cells represent a heterogeneous cell population containing haematopoietic stem and progenitor cells. These cells have been identified as potential candidates for use in cell therapy for the regeneration of damaged tissues caused by trauma, degenerative diseases, ischaemia and inflammation or cancer treatment. In our study, we examined a model using whole-body irradiation and the transplantation of bone marrow (BM) or haematopoietic stem cells (HSCs) to study the repair of haematopoiesis, extramedullary haematopoiesis and the migration of green fluorescent protein (GFP⁺) transplanted cells into non-haematopoietic tissues. We investigated the repair of damage to the BM, peripheral blood, spleen and thymus and assessed the ability of this treatment to induce the entry of BM cells or GFP⁺lin⁻Sca-1⁺ cells into non-haematopoietic tissues. The transplantation of BM cells or GFP⁺lin⁻Sca-1⁺ cells from GFP transgenic mice successfully repopulated haematopoiesis and the haematopoietic niche in haematopoietic tissues, specifically the BM, spleen and thymus. The transplanted GFP⁺ cells also entered the gastrointestinal tract (GIT) following whole-body irradiation. Our results demonstrate that whole-body irradiation does not significantly alter the integrity of tissues such as those in the small intestine and liver. Whole-body irradiation also induced myeloablation and chimerism in tissues, and induced the entry of transplanted cells into the small intestine and liver. This result demonstrates that grafted BM cells or GFP⁺lin⁻Sca-1⁺ cells are not transient in the GIT. Thus, these transplanted cells could be used for the long-term treatment of various pathologies or as a one-time treatment option if myeloablation-induced chimerism alone is not sufficient to induce the entry of transplanted cells into non-haematopoietic tissues.     

Characterization of the interactions between stromal and haematopoietic progenitor cells in expansion cell culture models

Development of the long-term culture models of haematopoietic stem cells (HSCs) is one of the important tasks in modern biotechnology. It has been suggested that stromal presence is important for haematopoiesis in vitro and in vivo, but the question remains: whether diffusible factors produced by stromal cells are sufficient for the regeneration of primitive and definitive haematopoietic cells, or direct cell-to-cell contacts of the cultured material with underlying stromal base would be required. During present studies, influence of various feeder layers and feeder layer conditioned media on proliferative, differentiative and clonogenic activity of human AC133+ derived from human umbilical cord blood was investigated. Cell extracts for feeder layers were prepared from 4-6 weeks old human embryos and co-cultured feeder cells. Effects of the conditioned media were also determined. Culture and feeder layer media were additionally supplemented with commonly implemented factors such as GM-CSF, IL-3 and LIF. Estimation of morpho-functional properties of AC133+ cultivated suspension cultures was performed in subculture experiments using semisolid agar culture conditions. Multipotential CFU-MIX (CFU-GEMM) and unipotential progenitor cells CFU-GM, BFU-E and CFU-E were observed and analyzed. Our data suggest that haematopoiesis can be sustained for prolonged cultivation periods in the presence of feeder layer cells or conditioned media supported culture models. Prolonged support of primitive haematopoietic cells and their clonogenic capacity and functional characteristics in feeder layer positive cultures, indicates that diffusible factors are sufficient for haematopoiesis and suggests that direct cell-to-cell contacts may not be exclusively required for successful long-term in vitro haematopoiesis.     

Drosophila haematopoiesis

Like in vertebrates, Drosophila haematopoiesis occurs in two waves. It gives rise to three types of haemocytes: plasmatocytes (phagocytosis), crystal cells (melanization) and lamellocytes (encapsulation of parasites). A first population of haemocytes, specified during embryogenesis, gives rise to an invariant number of plasmatocytes and crystal cells. A second population of haemocytes is specified during larval development in a specialized haematopoietic organ, the lymph gland. All three types of haemocytes can be specified in this organ, but lamellocytes only differentiate in response to parasitism. Thus, larval in contrast to embryonic haematopoiesis can be modulated by physiological constraints. Molecular cascades controlling embryonic haematopoiesis are relatively well established and require transactivators such as GATA, FOG and Runx factors, which are also co-opted in mammalian haematopoiesis. Mechanisms involved during larval haematopoiesis are less well understood although a number of chromatin remodelling factors and signalling pathways (JAK/STAT, Toll, Hedgehog, Notch) are required. In healthy larvae a pool of progenitors is maintained within the lymph gland, under the control of a signalling centre which expresses Collier, Serrate, Antennapedia and Hedgehog, and controls haemocyte homeostasis. Its key role in haemocyte homeostasis is reminiscent of interactions described in vertebrates between haematopoietic stem cells and their microenvironment (niche).     

昆虫造血作用和造血干细胞研究进展

昆虫血细胞（insect hemocyte）在昆虫代谢、 发育变态以及先天免疫等方面承担着重要的作用。昆虫只有先天免疫系统, 血细胞所行使的免疫功能对于昆虫对抗外源病菌尤为重要。本文主要介绍了昆虫血细胞类型、 造血作用、 造血干细胞及造血相关因子的相关研究。通过特殊染色和形态学观察, 果蝇Drosophila血细胞主要由3类细胞组成, 而鳞翅目等大部分昆虫血细胞由5类细胞组成。昆虫血细胞主要存在于循环血液环境及造血器官内, 而在这两个系统中都存在有进行复制的血细胞, 这为研究昆虫造血干细胞特性和其定位提供了一个很好的系统。果蝇血细胞祖细胞来自于胚胎中胚层细胞, 然后再分化为各种血细胞, 这一系列分化过程由造血因子所调控。     

鳗鲡出血性开口病的病理学研究

本文报告了我国发生的一种鳗鲡病毒病──鳗鲡出血性开口病的组织病理变化：肝、肾、脾脏组织出血、细胞变性,骨质内有大县白细胞浸润,肝、肾、脾脏细胞超微结构病理变化明显,肾、脾脏造血组织和外周血细胞出现核染色质边集、奇异形核,大量髓鞘样结构、自噬体和自噬溶酶体等,并可见类似凋亡细胞及调亡小体结构和邻近细胞内吞噬体增多现象。根据骨组织中白细胞浸润及器官和血液中部分细胞结构已出现异型性特征,作者认为,鳗鲡出血性开口病可能有癌变的趋势．     

EVALUATION OF THE SPECIFICITY OF A LYMPHOID CHALONE

An extract from calf spleens, injected into mice, was found to inhibit their lymphocyte proliferative response to PHA-M and PWM in vitro. Despite the ability of the spleen extract to inhibit the response of lymphocytes to stimuli in vitro, no effect was observed on the repopulation of lymphocytes in the peripheral blood of sublethally irradiated mice. The data suggest that the spleen extract acts as a specific inhibitor of the immune competent cells since neither the precursors of lymphocytes nor other haematopoietic cells were affected.     

Recapitulative haematopoietic development of human pluripotent stem cells in the absence of exogenous haematopoietic cytokines

To improve the recapitulative quality of human pluripotent stem cell (hPSC) differentiation, we removed exogenous haematopoietic cytokines from the defined differentiation system. Here, we show that endogenous stimuli and VEGF are sufficient to induce robust hPSC-derived haematopoiesis, intensive generation of haematopoietic progenitors, maturation of blood cells and the emergence of definitive precursor cells including those that phenotypically identical to early human embryonic haematopoietic stem cells (HSCs). Moreover, the cytokine-free system produces significantly higher numbers of haematopoietic progenitors compared to the published protocols. The removal of cytokines revealed a broad developmental potential of the early blood cells, stabilized the hPSC-derived definitive precursors and led to spontaneous activation of inflammatory signalling. Our cytokine-free protocol is simple, efficient, reproducible and applicable for embryonic stem cells (ESCs) and induced PSCs. The spectrum of recapitulative features of the novel protocol makes the cytokine-free differentiation a preferred model for studying the early human haematopoietic development.