妊娠期糖尿病胎盘SHBG的免疫细胞化学与酶标免疫电镜观察

目的探讨GDM时胎盘滋养细胞存在SHBG及其变化与GDM发病机制的关系。方法应用免疫细胞化学染色以及包埋前免疫电镜技术,检测GDM胎盘组织中SHBG的表达和观察SHBG在GDM胎盘组织细胞内的分布特征。结果电镜下GDM组胎盘合体滋养细胞表面微绒毛排列紊乱、肿胀、数目减少,绒毛间隙变窄、断裂。胞质内粗面内质网扩张,线粒体肿胀,细胞核形状不规则,染色质分布异常,未见高电子密度的颗粒。光镜下GDM组免疫阳性染色分布于整个胎盘合体滋养细胞,主要位于细胞膜和细胞浆,但是微绒毛侧和基底膜侧着色都不甚均匀,表达弱。GDM组胎盘SHBG免疫染色阳性表达率(25/30,83.3%)较对照组胎盘SHBG免疫染色阳性表达率(28/30,93.3%)减低(P0.01)。结论 GDM时胎盘滋养细胞存在着SHBG,但其合成和分泌减少,在GDM发病过程中的发挥着重要作用。     

胎盘部位滋养细胞肿瘤的临床病理特点及鉴别诊断

目的了解胎盘部位滋养细胞肿瘤（ＰＳＴＴ）的临床病理特点及其鉴别诊断要点。方法对５例ＰＳＴＴ进行临床,光镜,电镜及免疫组化研究,并同时对比观察１０例绒癌及２例过度的胎盘部位反应（ＥＰＳ）。结果ＰＳＴＴ见于育龄妇女,前次妊娠多为足月产,常见症状为闭经及／或阴道出血。血清绒毛膜促性腺激素（ｈＣＧ）可有轻～中度升高。光镜下瘤细胞组成单一,仅见中间型滋养细胞,成片状,条索状穿插浸润于子宫平滑肌束间。血管浸润明显,却少有出血或坏死。分裂相少见。电镜下瘤细胞核周有丰富的中间丝。免疫组化表现为胎盘催乳素（ｈＰＬ）阳性而ｈＣＧ多为阴性。结论ＰＳＴＴ为一少见的滋养细胞肿瘤,具有独特的光镜,电镜及免疫组化特点,这些特点可与其它滋养细胞疾病鉴别     

胎盘合体滋养细胞的免疫抑制活性

<正> 选用正常足月妊娠新鲜胎盘,经酶消化合并机械分离法分离胎盘合体滋养细胞。所获得的合体滋养细胞的纯度达90～95％以上,细胞活力大于90％。经培养4小时的合体滋养细胞的上清液,对母亲与胎儿的淋巴细胞转化试验和向混合淋巴细胞培养均有明显的抑制。提示可能通过局部抑制母体免疫     

鼠心房起搏样细胞的光镜和电镜观察

目的探讨大鼠和小鼠心房传导组织以外的起搏样细胞。方法应用石蜡、冰冻及超薄切片，采用HE、Masson和胆碱脂酶组织化学染色，分别在光镜和电镜下观察大鼠和小鼠的心房组织，寻找心房组织中隐匿的起搏细胞样细胞。结果右心房肌层内散在、无规律地存在一些圆形或卵圆形细胞，这些细胞核相对较大，细胞质清亮，细胞器较少。它们具备窦房结内起搏(P)细胞的特征。结论大鼠和小鼠左、右心房组织内存在起搏细胞样细胞，它们很可能是心房异位搏动的形态学基础。     

小鼠早中期胚胎组织中RNA结合基序蛋白10、caspase-9和内皮型一氧化氮合酶蛋白的表达

目的:观察小鼠早中期胚胎组织中RNA结合基序蛋白10(RBM10)、caspase-9和内皮型一氧化氮合酶(eNOS)的表达和定位,初步探究三者相关性。方法:取7.5～9.5d孕鼠胚胎包埋切片,行免疫组织化学法检测RBM10、caspase-9和eNOS的表达。结果:免疫组织化学结果显示,RBM10主要表达于7.5d的胚胎细胞核、原始消化管细胞核内,8.5 d的胚胎各胚层细胞核内(头部与体壁细胞阳性表达显著)、滋养层及胚外中胚层细胞核内,9.5 d的合体滋养层细胞核、部分胚胎上皮细胞细胞核;且在7.5、8.5 d和9.5 d的蜕膜细胞核均有阳性表达。Caspase-9主要表达于7.5 d的靠近体蒂位置处羊膜上皮、胚胎体壁细胞胞质,8.5 d的胚胎羊膜上皮胞质,9.5 d的胚胎原始消化管、原始心壁等上皮细胞胞质。eNOS主要表达于7.5、8.5、9.5 d的滋养层毛细血管内皮细胞胞质,另外在8.5 d的合体滋养层内侧即胚外中胚层细胞之间的毛细血管内皮可见阳性表达。各组8.5 d、9.5 d RBM10、caspase-9和eNOS表达差异均有统计学意义。结论:RBM10可能在促进血管形成方面与eNOS有相关性。     

恶性血管周上皮样细胞肿瘤2例并文献复习

目的 探讨恶性血管周上皮样细胞肿瘤(malignant perivascular epitheliod cell tumor,PEComa)的临床、病理特征及鉴别诊断.方法 对2例恶性PEComa进行临床、病理形态学、免疫表型及电镜观察,并复习相关文献.结果 镜下见肿瘤组织由梭形细胞及上皮样细胞构成,细胞异型性明显,细胞胞质透明或嗜酸,细胞核大,有核仁,瘤细胞由纤细的纤维结缔组织及薄壁血管将其分隔成巢状、腺泡状、片状结构,部分区域可见多核瘤巨细胞伴肿瘤坏死,核分裂多见.免疫表型:肿瘤细胞表达HMB-45、S-100、SMA及desmin;电镜下见肿瘤细胞胞质内可见高电子密度核心的内分泌小体和少许不成熟的黑色素小体.结论 恶性PEComa罕见,病理形态多样,熟悉其临床、病理形态、免疫表型及电镜下改变,有助于临床、病理医师对其正确诊断及鉴别诊断.     

Bcl—2蛋白在正常、妊高征及胎儿宫内发育迟缓胎盘中的表达

目的 通过检测Bcl-2蛋白在正常各期胎盘、妊高征胎盘和胎儿宫内发育迟缓胎盘中的表达，探讨Bcl-2蛋白在胎盘的发育过程中的功能及妊高征和胎儿宫内发育迟缓的病理机制。方法 取正常的8－10周、20－22周、足月胎盘以及妊高征和胎儿宫内发育迟结的胎盘组织固定，石蜡包坦，用ABC法抗Bcl-2免疫组织化学染色，光镜观察。结果 正常早孕绒毛合体细胞滋养怪、细胞滋养层细胞核、绒毛吵轴细胞阳性，细胞滋养层的胞质、绒毛中轴基质阴性。正常中期和足月胎盘滋养层细胞、血管内皮细胞阳性。与正常足月胎盘相比，妊高征、台儿宫内发育迟缓胎盘几科不着色。结论 Bcl-2蛋白在正常各期胎盘中均有表达，而在妊高征和胎儿宫内发育迟缓胎盘几乎不表达，Bcl-2蛋白表达异常与妊高征与胎儿宫内发育迟缓的发病机制有关。     

对正常人胎盘绒毛冷冻复型电镜研究

本文应用冷冻复型电境技术对22例由40天到足月胎儿胎盘绒毛的超微结构进行了研究,发现妊娠早期合体细胞层较厚,游离面有大量细长微绒毛,胞质内含丰富管泡,随妊娠进行,微绒毛逐渐减少变短,末端扩张,胞质内大泡逐渐增多。细胞核孔随胎龄增长而增多。在合体细胞近游离端还发现有紧密连接和不完全分隔合体细胞的膜。细胞滋养层细胞早期成层,以后逐渐减少,细胞外形规则,细胞器少,核孔变化与合体细胞者基本相同,细胞间及与合体细胞间可见桥粒与缝管连接。在二层细胞间我们观察到由细胞滋养层向合体细胞过渡分化的细胞。对上述结果的意义进行了讨论。     

人早期胎盘绒毛运动神经诱向因子1及其受体的免疫组织化学定位

为了解人胎盘绒毛是不存在运动神经诱向因子及其可能的功能意义，本实验用ＭＮＴＦ１单克隆抗体及抗独特型单克隆抗体在人早期胎盘绒毛石蜡切片上进行免疫组织化学反应，对人早期胎盘绒毛的ＭＮＴＦ１及其受体进行定位。结果显示，胎盘绒毛的细胞滋养层细胞，合体滋养层细胞和基质细胞均呈ＭＮＴＦ１强免疫反尖，ＭＮＴＦ１样免疫反应物质分布在胞质内，胞核内阴性。     

大鼠卵泡膜外层平滑肌细胞的超微结构研究

成年雌性大鼠重约150～200g,共30只。检查阴道涂片,选动情期处死。一侧卵巢作电镜观察,对侧卵巢作石蜡切片,供光镜观察。生长卵泡和囊状卵泡的卵泡壁均有卵泡膜,其内、外层分界不清。电镜下可辨认分泌细胞、成纤雏细胞和平滑肌细胞。分泌细胞位于卵泡膜内层,呈梭形或卵圆形,胞质内有滑面内质网、线粒体和脂滴;成纤维细胞广泛存在于卵泡膜的内、外层,细胞呈长梭形,细胞核较长,胞质内富有粗面内质网、游离核蛋白体和线粒体;平滑肌细胞在卵泡膜外层,按照A. P. Somlyo和A. V. Somlyo的识别标准,它具有典型的平滑肌细胞超微结构特征:细胞膜凹入的吞饮小泡;胞质内有纵行排列的肌丝;与肌丝相连的密体;线粒体和内质网集中在核极区。本文观察的成年大鼠囊状卵泡的卵泡膜外层在动情期确有典型的平滑肌细胞。文中还讨论了卵泡膜平滑肌细胞的超微结构、细胞分化和存在等问题。     

Immuno-electronmicroscopic identification and localization of the antigenic proteins of tree pollen grains

The localization of antigenic proteins on ultrathin sections of pollen grains represents an interesting approach to understanding the release mechanisms of these antigens when the pollen grains come in contact with various physiological fluids. Using different rabbit antibodies we have demonstrated the locations of these antigens in the various structures of pollen grains. We further demonstrated the cross-reactivities between alder (Alnus incana), birch (Betula verrucosa) and hazel (Corylus avellana) pollen allergens. Ultrathin sections of the pollen grains were prepared and allowed to react with two individually raised rabbit antibodies, (Ab-BV and Ab-ALK), against birch pollen. The sites of the Ag/Ab complex on the sections were labelled by protein A/gold, and identified in a transmission electron microscope. The two birch antibodies showed either quantitative or qualitative differences regarding their binding to various structures on the pollen sections. Using Ab-BV, the antigen-binding sites were located in the apertural region of the pollen grain and in the cytoplasm, while almost no gold labelling could be seen on the pollen surface. With the other antibodies (Ab-ALK), we could visualize the antigen-binding locations on the surface material of the pollen grains, particularly in the exine part of the wall and in the cytoplasm. A few gold particles could also be seen in the apertural region of the pollen. In hazel and alder pollen the exine part of the wall was the most densely labelled, whereas the cytoplasm and the aperture bound smaller numbers of gold particles. Cross-incubations: birch pollen incubated with antibodies against hazel (Ab-CA), or alder (Ab-AI), showed various intensities of gold labelling for each of the three species. Statistically, the differences in the number of gold particles bound per micron 2 grain section between birch, hazel and alder, were highly significant. The cross-reactivities between these antigens from the three pollen species were further tested using house-produced rabbit antisera against antigens of the three species by means of electrophoretic and autoradiographic techniques (CIE and CRIE). The three antibodies could precipitate the major IgE-binding antigen from all three pollen species.     

Preliminary investigations of a correlation between electron energy loss and morphometric analyses on ultrathin cryosections from normal and neoplastic gastric tissues

Summary Electron energy loss spectroscopy (EELS) has been used to measure the ratios of C, N, O, P and Ca in ultrathin cryosections from normal and neoplastic gastric tissues. First results show a correlation between the EELS, and morphometric data in cells from these tissues. We have found that ultrathin, freeze-dried cryosections, with an average thickness of up to 75 nm, are stable enough for EELS-analysis in a 200 KV electron microscope with an adapted Gatan-EELS-Spectrometer.Supported in part by Deutsche Forschungsgemeinschaft Wo 343/1-1 and Fraunhofer-Gesellschaft Wo 3/7 P 3404937.1     

Detection of UCH-L1 expression by pre-embedding immunoelectron microscopy with colloidal gold labeling in diseased glomeruli

The purpose of this report was to study the use of pre-embedding immunoelectron microscopy technique with gold and horseradish peroxidase (HRP) labeling in detecting the expression of ubiquitin C-terminal hydrolase 1 (UCH-L1) of podocytes in glomerulonephritis. The specimens of human IgA nephropathy and lupus nephritis were fixed with paraformaldehyde and lysine-HCl buffer, labeled by colloidal gold or HRP, embedded with epoxy resin, and examined under the transmission electron microscope. The high density of gold particles or peroxidase reaction products (DAB) combined with UCH-L1 was obvious in cytoplasm and processes of podocytes. This modified technique of pre-embedding immunoelectron microscopy could perfectly preserve the ultrastructure of kidney and expose antigens which is valuable for clinical diagnostic work and experimental research.     

Study of the cultured human endothelial cells infected by epidemic hemorrhagic fever virus]

Virus antigen could be detected in the cytoplasm of infected human endothelial cells (HEC) by immunofluorescent assay (IFA) 2 to 10 days after the inoculation of epidemic hemorrhagic fever virus (EHFV), but no apparent histologic changes could be found by phase contrast light microscopy, as well as no mature virus particles could be detected under the transmission electron microscope. Reinoculation of the freeze-melt supernatant of HEC 8 days after the inoculation of EHFV to EHFV susceptible Vero E-6 cells, viral antigen could be detected in most of these cells and mature EHFV particles or viral inclusion bodies could also be obtained in the cytoplasm under transmission electron microscope. The results show that HEC is a susceptible target cell to EHFV and infection by this virus may not give apparent cytopathogenic effect in HEC.     

兔抗脱氢表雄酮抗体的鉴定及脱氢表雄酮在人胎盘的定位

为了鉴定脱氢表雄酮抗体的特异性及使用效价,观察脱氢表雄酮在人早期胎盘绒毛的细胞定位。方法：免疫组织ＡＢＣ法。结果：脱氢表酮抗体烃度为１：１０００～１：２０００时,胎盘切片仍显示较强的脱氢表雄酮免疫反应阳性,抗体经过量的脱氢表雄雄酮－ＢＳＡ偶联复合物吸收后,再进行孵育时,胎盘切片呈阴性反应,提示该脱氢表雄酮本有较高的使用效价及特异性。人胎盘绒毛两层滋养层细胞和基质细胞均早脱氢表雄酮免疫反应性,阳性反     

Immunoelectron microscope observations on secretion of human placental lactogen (hPL) in the human chorionic villi

Immunoelectron microscope localization of human placental lactogen (hPL) was investigated in the chorionic villi from week 7 to full-term gestation with the protein A-gold technique. With specific antiserum against hPL, immuno-reactive gold particles were found to be preferentially located in Golgi-derived, electron-dense small granules of 80-180 nm in the syncytiotrophoblast. Our electron micrographs indicate that these small granules increase in number in the course of gestation and are released by exocytosis from the apical cell surface. The present study reveals that hPL is segregated from the Golgi apparatus, stored in the syncytiotrophoblast as secretory granules, and released into the maternal blood.     

Cytomegalovirus infection of the human placenta: an immunocytochemical study.

In congenital cytomegalovirus (CMV) infection histologic evaluation of the placenta is often unrevealing. In the present study immunocytochemistry to CMV immediate early and early nuclear antigens was used to characterize placental involvement in six cases of symptomatic intrauterine CMV infection. Histologic examination had demonstrated diagnostic viral inclusions in one placenta and non-specific villitis in another. However, immunocytochemistry revealed CMV infection in five of the six placentas, including three with no pathologic changes on routine histologic evaluation. Infected cells were located primarily in the villous stroma. In one case immunoperoxidase staining showed infection in the syncytiotrophoblast. Infected endothelial cells were demonstrated by double staining for CMV and factor VIII antigen. No double-stained cells were seen in tissue sections stained for CMV immediate early nuclear antigen or the human macrophage-associated CD68 antigen, which is expressed in Hofbauer cells. In conclusion, specific immunoperoxidase staining was more sensitive for demonstrating placental CMV infection than was histologic examination and it aided in the characterization of infected cells.     

Significance and value of immunohistochemical localization of pregnancy specific proteins in feto-maternal tissue throughout pregnancy

Tissue from 50 cases of products of conception and placenta at different gestational ages from as early as 12 d post ovulation up to 40 wk were examined by immunoperoxidase technique for localization of HCG, HPL, SP1, and PAPP-A. HCG was localized in the cytoplasm of syncytiotrophoblast (ST) with strong intensity in the 12-d blastocyst and remained strong until 8 to 10 wk. It then gradually decreased, becoming almost negative in term placenta. HCG was also seen in the cytoplasm of intermediate trophoblast (IT) at the implantation site but with variability in staining. HPL and SP1 appeared later than HCG in ST, and the intensity of staining increased rapidly to strongly positive by wk 8. They remained strong until full term. IT stained strongly positive with HPL throughout pregnancy, and some were different from the HCG positive ones. ST were constantly negative for PAPP-A throughout pregnancy. The latter, however, was definitely seen in the cytoplasm of cytotrophoblast (CT) of early blastocyst, the superficial epithelium of the endometrium adjacent to the implantation site, in many decidual cells around the implantation site and in the amniotic membrane epithelium.     

人胎盘脱氢表雄酮的免疫组织化学研究

目的　观察脱氢表雄酮 (DHEA)在人胎盘的定位及其含量的周龄变化。　方法　用免疫组织化学及定量方法。　结果　人胎盘两层滋养层细胞和基质细胞均呈 DHEA免疫反应性 ,反应物质分布于胞质 ,胞核无免疫反应。在人胎盘绒毛的不同发育阶段中 ,DHEA的相对含量随周龄的增加而增加。　结论　 DHEA对胚胎及胎盘的发育可能具有重要的调节功能。     

人胎盘来源的间充质干细胞向血管内皮细胞分化潜能的研究

目的: 探讨人胎盘来源的间充质干细胞(placenta derived mesenchymal stem cells,PDMSCs)在体外分化为血管内皮细胞的潜能。方法: 利用组织块种植法体外分离培养PDMSCs,流式细胞术鉴定其表面抗原,应用含50 μg/L VEGF和10 μg/L bFGF的诱导分化液定向诱导PDMSCs向内皮细胞分化。免疫细胞化学检测分化过程中不同时点内皮特异性标志的表达变化。透射电镜和体外血管生成实验分别检测内皮特异性结构和内皮细胞功能。结果: PDMSCs表面抗原 CD105和CD166阳性,CD31、CD34和CD45阴性。诱导分化的内皮细胞形态呈鹅卵石样,早期内皮标志Flk-1/KDR在4 d表达最强;成熟的内皮标志CD31、vWF、CD144/VE-cadherin在内皮细胞分化过程中呈现时间依赖性表达(0 d、4 d、8 d和12 d)。透射电镜下可见内皮特异性结构:Weibel-Palade小体。细胞接种在细胞外基质凝胶中可形成毛细血管样结构。结论: 胎盘中可分离出大量PDMSCs,并且PDMSCs在体外可分化为具有功能特性的内皮细胞,因此胎盘组织可能成为血管组织工程和再生医学的理想种子细胞来源。