纤维粘连蛋白和层粘蛋白对培养血管平滑肌细胞生长和分裂的影响

我们用不同剂量的纤维粘连蛋白（Ｆｎ）和层粘连蛋白（Ｌｎ）作用于培养的血管平滑肌细胞（ＶＳＭＣｓ）观察ＶＳＭＣｓ的生长和分裂情况，发现Ｆｎ和Ｌｎ均能促进ＶＳＭＣＳ的生长的分裂，Ｆｎ的这种作用在４０μｇ以上呈剂量依赖性，Ｌｎ则在２４μｇ以下呈剂量依赖性，当细胞数达到最大后，Ｆｎ和Ｌｎ的促进作用均逐步减弱。     

纤维粘连蛋白和层粘连蛋白对培养血管平滑肌细胞生长和分裂的影响

我们用不同剂量的纤维粘连蛋白（Ｆｉｂｒｏｎｅｃｔｉｎ，Ｆｎ）和层粘连蛋白（Ｌａｍｉｎｉｎ，Ｌｎ）作用于培养的血管平滑肌细胞（ＶＳＭＣｓ）观察ＶＳＭＣｓ的生长和分裂情况，发现Ｆｎ和Ｌｎ均能促进ＶＳＭＣＳ的生长和分裂，Ｆｎ的这种作用在４０μｇ以下呈剂量依赖性，Ｌｎ则在２４μｇ以下呈剂量依赖性，当细胞数达到最大后，Ｆｎ和Ｌｎ的促进作用均逐步减弱。     

小鼠胸腺基质细胞系的分型

为鉴定８株小鼠胸腺基质细胞（Ｔｈｙｍｉｃｓｔｒｏｍａｌｃｅｌｌｓ，Ｔｓｃ）系的类型及本室研制出的抗小鼠ＴＳＣ单抗的价值，我们采用细胞ＥＬＩＳＡ和免疫荧光法对３０种自制的及国外引进的ＭＴＳ系列抗小鼠ＴＳＣ单抗与８株ＭＴＳＣ细胞系的反应性进行了观察，结果表明：①ＭＴＳＣ１，２为髓质型胸腺上皮细胞（Ｔｈｙｍｉｃｅｐｉｔｈｅｌｉａｌｃｅｌｌｓ，ＴＥＣ）的不同亚群；ＭＴＳＣ３，５，６，７，８为皮髓质交界型ＴＥＣ的不同亚群。②在不同类型的ＴＳＣ间及胸腺细胞和ＴＳＣ间，均存在共同抗原。③ＭＴＳ系列ＭｃＡｂ不能鉴定的ＭＴＳＣ系，能用本室自制的抗小鼠ＴＳＣＭｃＡｂｓ定型。     

MCP-1 mcAb和Losartan拮抗Ang Ⅱ介导的VSMCs的增殖与迁移效应的实验研究

目的探讨单核细胞趋化蛋白－１单克隆抗体（ＭＣＰ－１－ｍｅＡｂ）或Ｌｏｓａｒｔａｎ 拮抗血管紧张素Ⅱ（ＡｕｇⅡ）介导的血管平滑肌细胞（ＶＳＭＣｓ）增殖与迁移效应的可能性，为防治动脉硬化提供一定的理论依据。方法采用改良的Ｂｏｙｄｅｎ小室法检测 ＶＳＭＣｓ的迁移效应；以 ＭＴＴ法、~３Ｈ－ＴａＲ掺入法、~３Ｈ－脯氨酸标记法和ＶＳＭＣｓ计数评价ＶＳＭＣｓ的增殖效应。将培养ＶＳＭＣｓ分为 ５组； ２％胎牛血清（ ２％ＦＣＳ）组、 ＡｕｇⅡ组（其浓度为 １０－１０～１０－６ｍｏｌ／Ｌ）、Ｌｏｓａｒｔａｎ组（其浓度为 １０－７～１０－５ｍｏｌ／Ｌ）、ＭＣＰ－ｌｍｃＡｂ组（终浓度为 １０μｇ／ｍｌ）和阳性对照组（含５％的酵母多糖活化血清）。结果（１） ＡｕｇⅡ具有剂量依赖性的促进 ＶＳＭＣｓ的增殖与迁移效应；（２） ＭＣＰ－１ｍｃＡｂ或 Ｌｏｓａｒ－ｔａｎ能有效地拮抗ＡｕｇⅡ介导的ＶＳＭＣｓ增殖与迁移效应。结论 ＭＣＰ－１ｍｃＡｂ或Ｌｏｓａｒｔａｎ对动脉硬化性疾病可能具有较大的应用前景。     

甲状腺素体外对Graves病T、B细胞功能异常影响的研究

目的：探讨Ｇｒａｖｅｓ病（ＧＤ）患者异常升高的甲状腺激素与免疫异常之间的因果关系,和抗甲抗腺药治疗ＧＤ期间加用甲状腺素（Ｔ４）的作用方法：将ＧＤ患者外在单个核细胞（ＰＢＭＣ）在不同浓度的Ｔ４条件下外培养６ｄ后,检测Ｔ细胞亚群和增减上清中可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）及ＬｇＧ的含量。结果：与正常对照组相比培养后的ＧＤ患者ＰＢＭＣ中ＣＤ８＋Ｔ细胞亚群百分率明显减少（Ｐ〈０．０１）;培养上清中Ｓ     

骶髓后连合核接受盆内脏伤害性信息传入的形态学证明

为阐明投射至骶髓后连合核的盆内脏初级传入中是否含有传递伤害性信息成分，本研究综合运用特异性标记初级传入Ｃ纤维的ＢＳＩ－Ｂ４－ＨＲＰ跨节追踪技术，神经干局部涂抹Ｃ纤维毒素ＣａＰｓａｉｃｉｎ并结合ＳＰ免疫组化方法，研究了猫投射至骶髓后连合核的盆神经初级传入纤维中是否含有传递伤害性刺激的成分；同时观察了秋水仙素处理的骶２后根节内ＢＳＩ－Ｂ４标记的初级传入神经元与ＳＰ免疫阳性神经元的关系。结果如下：（１）向盆神经注入ＢＳＩ－Ｂ４－ＨＲＰ，骶１～３后根节内出现平均直径３４μｍ的标记细胞，后连合核内出现密集的标记终末，电镜下证明通过Ｌｉｓｓａｕｅｒ氏束进入脊髓内的标记纤维均为无髓纤维；（２）对盆神经进行局部Ｃａｐｓａｉｃｉｎ处理，引起后连合核内的ＳＰ免疫阳性纤维和终末明显减少；（３）骶２后根节内ＢＳＩ－Ｂ４－ＦＩＴＣ标记细胞有１７％同时呈ＳＰ免疫阳性；（４）骶２后根节内ＢＳＩ－Ｂ４－ＨＲＰ标记的盆内脏初级传人神经元的３９％同时呈ＳＰ免疫阳性。本研究结果在形态学上证实了骶髓后连合核接受盆腔内脏伤害性信息的传入，它可能是中继和整合盆内脏伤害性信息的低级中枢。     

PC和KC在贮脂细胞激活过程中的协同作用

近年来，对人和实验性肝纤维化发病机理的研究焦点主要集中于贮脂细胞（ＦＳＣ）的病理学改变，而有关正常或不完全损伤肝实质细胞（ＰＣ）在ＦＳＣ激活过程中的作用少见报道。本文对正常和ＣＣｌ４损伤大鼠ＰＣ进行分离培养，观察ＰＣ和库普弗细胞（ＫＣ）在ＦＳＣ激活过程中的协同作用。采用ＰＣ、ＫＣ和ＦＳＣ分离培养，用流式细胞仪测定ＤＮＡ含量，免疫组化和图像分析对ＦＳＣ中Ⅱ、Ⅲ、Ⅳ型胶原和ＦＮ进行定量分析，液闪法检测ＦＳＣ中３Ｈ－脯氨酸掺入量。来自正常或ＣＣｌ４损伤大鼠ＰＣ条件培养基（ＰＣｎｃｍ、ＰＣｔｃｍ）对原代培养ＦＳＣ的增殖具有较强的刺激效应，ＦＳＣ内３Ｈ－脯氨酸掺入量呈现时间和剂量相关性地升高，ＤＮＡ含量在时间和剂量上也呈正相关的增加。单独用ＰＣｎｃｍ或ＰＣｎｃｍ＋ＰＣｔｃｍ合用时，ＦＳＣ表现出明显地转化特征，而ＦＳＣ内Ⅰ、Ⅲ、Ⅳ型胶原和纤维粘连蛋白（ＦＮ）均显著升高。ＰＣ和ＫＣ对ＦＳＣ的激活具有协同效应，并呈现时空相关性，对ＦＳＣ内ＥＣＭ合成的协同效应与增加细胞总量及增加单个ＦＳＣ合成ＥＣＭ的能力有关。     

纤维肉瘤的诊断及鉴别诊断

目的：进一步探讨纤维肉瘤与其它梭形细胞软组织肿瘤的鉴别诊断。方法：分别选取１９８０年前（５２例）与１９８０年后（１８例）原病理诊断为纤维肉瘤病例,经３位病理医师按标准复查原理切片,结果一致者３０例,选出其中２０例和另外有争议的２４例,共４４例分别作免疫组织化学ｖｉｍｅｎｔｉｎ、α１－ＡＴ、Ｓ－１００、ＮＦ,ｄｅｓｍｉｎ、ｍｙｏｇｌｏｂｉｎ、Ｃ－ｋｅｒａｔｉｎ染色。结果：１９８０年前５２例中维持原诊     

胃肠道间质肿瘤组织发生的探讨

目的探讨胃肠道间质肿瘤（ｇａｓｔｒｏｉｎｔｅｓｔｉｎａｌｓｔｒｏｍａｌｔｕｍｏｒ，ＧＩＳＴ）的组织发生。方法对３６例ＧＩＳＴ进行组织学、组织化学及免疫组织化学观察。结果胃和大肠间质肿瘤呈间叶细胞表型，但光镜下观察到在１５例发生于小肠的梭形细胞肿瘤中，瘤细胞之间存在着嗜酸性小体即丝团样纤维（ｓｋｅｉｎｏｉｄｆｉｂｅｒ，ＳＦ），Ｍａｓｓｏｎ三色染色呈蓝色，ＰＡＳ呈强阳性；免疫组化染色显示Ｓ－１００蛋白阳性率为４４．４４％，ｄｅｓｍｉｎ阳性率为８．３３％，瘤细胞的免疫表达与其分化程度无关。结论胃和大肠的间质肿瘤免疫表型复杂，提示其起源与原始间叶细胞有关，而小肠间质肿瘤大多数为神经源性肿瘤。可能起源于肠壁内在神经丛的神经膜细胞或来自小肠的自律神经。     

树突细胞从凋亡癌细胞获取抗原后的免疫应答

目的 观察树状细胞（ＤＣｓ）从凋亡胆管癌细胞获取原后，体外诱导抗肿瘤免疫应答及对胆管的特异性免疫杀伤效果，方法 用粒－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）加白介素－４（ＩＬ－４）从人外周血分化、诱导ＤＣＳ，γ－射线在体外诱导培养的人胆管癌细胞凋亡，将ＤＣＳ、Ｔ淋巴细胞和凋亡胆管癌细胞共培养，同时设计不同类型肿瘤细胞（坏死胆管癌细胞及培养胆管癌细胞）作对照，７ｄ后，分离、富集ＤＣｓ、Ｔｉｓｓ ｃｎｈ     

The interaction of Schwann cells with chitosan membranes and fibers in vitro

The bridging of nerve gaps is still one of the major problems in peripheral nerve regeneration. A promising alternative for the repair of peripheral nerve injuries is the bioartificial nerve graft, comprised of a biomaterial pre-seeded with Schwann cells (SCs), which is an effective substrate for enhancing nerve regeneration. Interaction between cultured SCs and biomaterials is of importance. For the purposes of this study, culture systems of normal SCs were used. The biocompatibility of chitosan, including chitosan membranes and chitosan fibers, was evaluated in vitro. The growth of SCs was observed by light and scanning electron microscopy at regular intervals. SCs were identified by immunocytochemical staining and the viability of SCs was measured by MTT assay. The experimental results indicated that SCs could grow onto chitosan materials with two different shapes: spherical and long olivary. They contacted with the extensions. The long olivary cells inclined to encircle chitosan fibers up. It was also found that the cells on the chitosan fibers migrated faster than those on the chitosan membranes. There was a good biological compatibility between chitosan and SCs. Compared with the chitosan membranes, SCs migrated more easily onto the stereoframe of chitosan fibers. These studies contribute information necessary to enhancing our understanding of biocompatibility of chitosan.     

睾丸支持细胞促进体外培养神经元生长的研究

目的　探讨睾丸支持细胞 (SC)对体外培养神经元生长发育的促进作用。　方法　将SCs制备成饲养层和SCs条件培养液 (SCM)后与神经元共培养 ,观察培养细胞的存活数和突起数 ,并应用图像分析仪观察细胞的面积。　结果　SCs饲养层及SCM培养神经元的存活数、突起数和胞体面积明显高于对照组 (P <0 0 1)。　结论SCs对体外培养的神经元具有显著的促生长作用。     

缺氧对体外培养的雪旺细胞形态和迁移的影响及EGb 761抗缺氧研究

为了探讨缺氧对体外培养在三维结构上的雪旺细胞的影响和ＥＧｂ７６１ 对缺氧雪旺细胞的保护作用,将在聚酯纤维上迁移的ＳＤ 大鼠的雪旺细胞进行缺氧培养,抗缺氧组培液含ＥＧｂ７６１。正常组常规培养。结果显示,缺氧组：缺氧３ ｄ,细胞停止迁移,细胞体由梭形变成半球形或球形;缺氧５ ｄ,细胞开始死亡,从聚酯纤维上脱落。抗缺氧组：缺氧３ ｄ 和正常组比较,细胞迁移和形态无明显改变;缺氧５ ｄ,少数远离植块的细胞呈半球形和球形;缺氧７ ｄ,少量死亡细胞脱离聚酯纤维。结果表明：缺氧将引起雪旺细胞迁移能力和方式及形态的改变,最后导致细胞死亡;ＥＧｂ７６１ 对缺氧的雪旺细胞有明显的保护作用。     

A single bout of exercise activates skeletal muscle satellite cells during subsequent overnight recovery

Skeletal muscle satellite cell (SC) content has been reported to increase following a single bout of exercise. Data on muscle fibre type-specific SC content and/or SC activation status are presently lacking. The objective of the study was to determine the impact of a single bout of exercise on muscle fibre type-specific SC content and activation status following subsequent overnight recovery. Eight healthy men (age, 20 ± 1 years) performed a single bout of combined endurance- and resistance-type exercise. Muscle biopsies were collected before and immediately after exercise, and following 9 h of postexercise, overnight recovery. Muscle fibre type-specific SC and myonuclear content and SC activation status were determined by immunohistochemical analyses. Satellite cell activation status was assessed by immunohistochemical staining for both Delta-like homologue 1 (DLK1) and Ki-67. Muscle fibre size and fibre area per nucleus were greater in type II compared with type I muscle fibres (P < 0.05). At baseline, no differences were observed in the percentage of SCs staining positive for DLK1 and/or Ki67 between fibre types. No significant changes were observed in SC content following 9 h of postexercise, overnight recovery; however, the percentage of DLK1-positive SCs increased significantly during overnight recovery, from 22 ± 5 to 41 ± 5% and from 24 ± 6 to 51 ± 9% in the type I and II muscle fibres, respectively. No changes were observed in the percentage of Ki-67-positive SCs. A single bout of exercise activates both type I and II skeletal muscle fibre SCs within a single night of postexercise recovery, preceding the subsequent increase in SC content.     

Effects of the in vivo predegenerated nerve graft on early Schwann cell migration: Quantitative analysis using S100-GFP mice

In peripheral nerve transection injury, continuity of axons as well as that of the basal lamina is disconnected. In such case, migrating Schwann cells (SCs) would be the only axonal guidance at an early stage of regeneration. However, it takes a few days for the dedifferentiated SCs to start migration, while axonal growth begins a few hours after injury. Consequently, the axons without guidance extensively branch out and wander off at the lesion, resulting in aberrant reinnervation. Therefore, enhancing SCs migration could be an attractive therapeutic strategy. In this study, we investigated the effects of the in vivo nerve predegeneration on SC migration and the time course of these changes. In our analysis, we established a novel animal model by nerve transplantation from S100-GFP mice (in which SCs constitutively express green fluorescent protein driven by the S100B promoter), by which SC migration could be exclusively visualized. Our results showed that SCs acquire the maximal migration ability with 14-day predegeneration, but subsequently it gradually decreased. There was a correlation between the time course of the changes in SC migration and the number of activated macrophages. These findings suggest that using predegenerated nerve grafts in repairing the transected nerves could facilitate SC migration into the recipient nerve stump. This technique could be beneficial for early establishment of axonal guidance and possible functional improvement after transection injury.     

人胎儿房室结的细胞分类--图像分析定量研究

目的　探讨胎儿房室结内的细胞类型和大小。　方法　石蜡切片 ,H E染色 ,光镜观察胎儿心脏房室结的细胞形态、类型 ;采用HIPAS 10 0 0图像分析仪对其进行定量分析。　结果　胎儿房室结的细胞主要为亮细胞、暗细胞和移行细胞。亮细胞和暗细胞为纺锤形或椭圆形 ,少数多边形 ;移行细胞细短柱形。亮细胞和移行细胞的细胞核明显大于暗细胞。　结论　胎儿房室结的起搏细胞包括亮细胞和暗细胞 ,它们可能是一种细胞的不同功能状态     

ECM molecules mediate both Schwann cell proliferation and activation to enhance neurite outgrowth

Tissue engineering using a combination of biomaterials and cells represents a new approach to nerve repair. We have investigated the effect that extracellular matrix (ECM) molecules have on Schwann cell (SC) attachment and proliferation on the nerve conduit material poly-3-hydroxybutyrate (PHB), and SC influence on neurite outgrowth in vitro. Initial SC attachment to PHB mats was unaffected by ECM molecules but proliferation increased (laminin > fibronectin > collagen). SCs seeded onto ECM-coated culture inserts suspended above a monolayer of NG108-15 cells determined the effect of released diffusible factors. The effect of direct contact between the two cell types on ECM molecules was also investigated. In both systems SCs enhanced neurite number per cell and percentage of NG108-15 cells sprouting neurites. NG108-15 cells grown in direct contact with SCs had significantly longer neurites than those exposed to diffusible factors when seeded on laminin or fibronectin. Diffusible factors released from SCs cultured on ECM molecules appear to initiate neurite outgrowth, whereas SC-neuron contact promotes neurite elongation. SC proliferation was maximal on poly-D-lysine-coated surfaces, but these cells did not influence neurite outgrowth to the levels of laminin or fibronectin. This suggests that ECM molecules enhance cell number and activate SCs to release neurite promoting factors. Addition of ECM molecules to PHB nerve conduits containing SCs is likely to provide benefits for the treatment of nerve injuries.     

一种小鼠肿瘤相关膜蛋白单抗对DCS细胞生物学行为的影响

探讨一种小鼠肿瘤相关膜蛋白在ＤＣＳ细胞表达的意义。方法用该蛋白抗处理ＤＣＳ细胞,观察其对细胞生长速度,软琼脂克隆形成能力,粘附性,细胞运动能力,水解酶活性,侵袭性,体内转移率等生物学行为方面的影响。结论该细胞膜蛋白可能调控小鼠肿瘤细胞增殖能力。     

体外培养中少突胶质细胞在立体网架上的迁移

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