5—羟色胺受体和P物质受体在培养的人胎盘滋养层细胞的定位研究

目的：探讨培养的人胎盘绒毛滋养层细胞是否存在5－羟色胺受体（5－HTR）和P物质受体（SPR）,方法：采用细胞培养法和免疫细胞化学技术。结果：培养的滋养层细胞为5－HTR与SPR免疫阳性反应,免疫阳性物质位于胞浆内,胞核为阴性。结论：人胎盘滋养层细胞自身含有5－HTR和SPR,为在离体培养条件下研究5－HT和SP对胎盘激素的合成与分泌的调控作用提供了形态学依据。     

胎盘部位滋养细胞肿瘤免疫组织化学分析

目的 :探讨免疫组织化学在胎盘部位滋养细胞肿瘤 (PSTT)病理诊断中的意义。方法 :应用免疫组织化学检测 8例PSTT中HPL、HCG及PCNA表达 ,并与胎盘部位过度反应及绒毛膜癌比较。结果 :HPL染色 8例PSTT均为阳性 ,阳性细胞指数为 6 2 2 5 %± 10 95 % ,高于绒毛膜癌的 11 13%± 9 2 6 %和EPS的 40 30 %± 2 3 90 % (P <0 0 1,P <0 0 5 ) ;6例PSTT细胞HCG染色阳性 (75 % ) ,阳性细胞指数为 10 87%± 8 31% ,EPS为 2 5 0 %± 4 40 % ,绒毛膜癌为 5 3 0 0 %± 2 2 10 % ;PSTT增殖细胞指数为 31 2 5 %± 8 86 % ,EPS为 10 37%± 5 2 8% ,绒毛膜癌为 71 10 %± 5 12 %。PSTT的HCG和增殖细胞指数与EPS和绒毛膜癌相比差异均有显著性 (P <0 0 1)。结论 :免疫组织化学检测HPL、HCG及PCNA对PSTT的诊断和鉴别诊断具有重要意义 ,但需结合临床与常规病理形态     

妊娠合并胎儿发育迟缓胎盘滋养层细胞凋亡与p53表达增高的相关性研究

目的　研究先兆子痫或吸烟导致妊娠合并胎儿发育迟缓胎盘滋养层细胞凋亡与促凋亡蛋白 p5 3和 Bax的表达及抗凋亡蛋白 Bcl- 2的表达。方法　选择无妊娠合并症妇女胎盘 4例 (n=4 ) ,先兆子痫或吸烟导致妊娠合并胎儿发育迟缓妇女胎盘 7例 (n=7)。胎盘组织切片用 HE染色和末端脱氧核糖核酸转移酶标记法 (TU NEL)染色 ,及细胞角蛋白 18裂解产物检测滋养层细胞凋亡。 Western蛋白免疫印迹和免疫组化方法检测 p5 3表达 ,免疫印迹方法检测 Bas、Bcl- 2、Bak、Bcl- XL 的表达。结果　妊娠合并胎儿发育迟缓胎盘滋养层细胞凋亡比对照组增多 ,这…     

滋养层细胞侵入相关基因在先兆子痫胎盘中的表达

探讨与滋养层侵入有关的细胞外基质分子相关基因在先兆子痫胎盘中的表达，采用分别点样有220余种人细胞因子相关基因和人类激素相关基因cDNA片段的两款基因芯片，检测经过严格配伍的先兆子痫和正常胎盘组织的基因表达谱差异。结果显示：钙粘蛋白、胶原、整合素、选择蛋白等18种细胞外基质分子基因的表达在先兆子痫和正常胎盘组织间相差2倍以上，且全部表现为在先兆子痫胎盘中的表达增强。先兆子痫患者的胎盘组织中基质金属蛋白酶(MMP)-10、-13、-15和金属蛋白酶组织抑制因子(TIMP)-2、TIMP-3、纤溶酶原、纤溶酶原激活物等的表达均较正常者高。提示胎盘中细胞外基质分子及其降解酶基因表达异常可能与先兆子痫的病理发生关系密切。     

DC-SIGNR在人胎盘滋养层细胞中的表达及意义

目的:探讨DC-SIGNR在人胎盘绒毛组织中的定位及在体外培养滋养层细胞上的表达.方法:建立稳定的滋养层细胞原代培养体系,采用免疫组化单染及荧光双染的方法检测不同孕期正常胎盘绒毛组织及体外培养滋养层细胞上DC-SIGNR的表达.结果:DC-SIGNR主要表达于胎盘滋养层细胞、Hofbauer细胞及胎盘微血管内皮细胞的胞质及包膜,在原代培养的人绒毛膜滋养层细胞中的DC-SIGNR的表达与在体组织表达一致.结论:DC-SIGNR表达于不同孕期的胎盘组织及体外分离培养滋养层细胞,为研究滋养层细胞上该受体在宫内感染中的作用提供体外实验的细胞学基础.     

正常足月胎盘和胎儿宫内生长迟缓胎盘中滋养细胞的凋亡及相关基因表达

目的 比较正常足月胎盘与胎儿宫内生长迟缓胎盘（IUGR）中的滋养细胞凋亡指数,以及bcl-2和fas的表达,以探讨滋养细胞的凋亡与bcl-2和fas表达的关系。方法 取正常足月胎盘组织和I－UGR胎盘组织各5例,固定、石蜡包埋、切片,每组分别TUNEL法以及ABC法抗bcl－2和抗fas免疫组织化学染色。镜下计数滋养细胞的凋亡指数,并对bcl-2和fas的表达量进行显微图像分析。结果 与正常足月胎盘相比,IUGR胎盘的凋亡指数显著增多（P＜0．01）,bcl－2表达量显著减低（P＜0．01）,而fas表达量无显著差异（P＞0．05）。结论 IUGR胎盘滋养细胞的凋亡指数明显增多可能与细胞bcl-2表达量减少有关,而不受fas表达量的影响。     

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     

FAS蛋白在子痫前期胎盘中的表达及意义

目的检测子痫前期胎盘的FAS蛋白表达水平,探讨子痫前期的发病机制。方法采用免疫组织化学方法结合病理图像分析技术,定量分析10例正常妊娠组、17例轻度子痫前期组和13例重度子痫前期组胎盘FAS蛋白表达。结果 FAS蛋白主要表达与胎盘绒毛滋养细胞的胞膜及胞浆内。与正常妊娠组比较,子痫前期组胎盘FAS蛋白表达显著增加,轻度、重度子痫前期组比较,有显著性差异。结沦子痫前期患者胎盘滋养层细胞FAS蛋白表达增加可能是子痫前期的发病机制之一,从而为临床治疗子痫前期提供新的方向。     

基质金属蛋白酶-9及其组织抑制物-1在人胎盘的表达及意义

目的研究基质金属蛋白酶-9(matrix metalloproteinases,MMP-9)及其组织抑制物-1(tissue inhibitor of metalloproteinases,TIMP-1)在不同孕期胎盘中的表达及与滋养层细胞侵蚀、子宫-胎儿血管系统建立的关系.方法应用原位杂交法检测56例胎盘(早孕16 例、中孕20 例、晚孕20例)中MMP-9及TIMP-1mRNA的表达.结果 MMP-9及TIMP-1mRNA主要表达于滋养细胞、绒毛间质血管壁及蜕膜组织中;MMP-9mRNA的表达早、中孕组明显强于晚孕组,TIMP-1mRNA的表达晚孕组明显强于早、中孕组,差异均有极显著性(P<0.01).结论 MMP-9及TIMP-1协同表达可能在滋养层细胞侵蚀、孕卵着床、血管重建过程中发挥一定的作用.     

探讨人胎盘泌乳素(HPL)标记中间滋养细胞在宫内外妊娠中的价值

目的:探讨HPL标记中间滋养细胞的诊断价值,籍以协助诊断宫内孕或宫外孕。方法:1组:选择蜕膜中疑似中间滋养细胞植入标本35例;Ⅱ组:宫内膜间质细胞肥硕呈蜕膜样变及/或宫内膜腺上皮呈A-S现象标本30例。对照组:宫内孕标本30例。对上述二组应用免疫组化SP法测定HLP,试剂选自福州迈新公司,染色结果以阳性细胞百分比及着色深浅综合判定。结果:对照组30例宫内孕具备蜕膜及绒毛,有24例HPL( ),6例(-)。Ⅰ组35例蜕膜组织疑似中间滋养细胞(IT)植入,28例HPL( ),证实为宫内孕;7例(-),证实2例为宫内孕,5例为宫外孕。Ⅱ组30例蜕膜样组织伴或不伴A-S现象,其中HPL 22例( )均为宫内孕,8例(-),证实6例为宫外孕,2例为宫内孕。结论:1组和2组HPL( )测定种植型中间滋养细胞对诊断宫内孕发生率高达50/65例(76.9%)及宫外孕11/65例(16.9%),HPL(-)11/15例(73.3%)为宫外孕及4/15(26.7%)为宫内孕,具有重要的临床诊断价值。     

Expression and subcellular localization of syntaxin 11 in human neutrophils

Objective  Syntaxin 11 mutations lead to familial hemophagocytic lymphohistiocytosis (FHL), characterized by uncontrolled hyperinflammation. This study examines the expression and subcellular localization of syntaxin 11 in human neutrophils as major inflammatory cells. Materials  The materials included human peripheral blood neutrophils, HL-60 cells. Methods  The methods used were RT-PCR, Western blot, immunocytochemistry, subcellular fractionation, HL-60 cell differentiation. Results  We have found that human peripheral blood neutrophils express syntaxin 11 mRNA and protein. Syntaxin 11 was upregulated during neutrophil differentiation of HL-60 cells. Syntaxin 11, identified as a membrane-bound protein, was broadly located in the plasma membrane and granules, with a predominant location in azurophilic granules of resting human neutrophils. A secondary location of syntaxin 11 was in specific and tertiary granules, which resulted in translocation to the plasma membrane on cell activation conditions that promoted the release of these organelles. Conclusions  These data indicate that human neutrophils express syntaxin 11 and call attention to the possible involvement of neutrophils in familial hemophagocytic lymphohistiocytosis pathology.     

人胆酸转运蛋白基因的克隆、亚细胞定位及其在肾组织中的表达

目的：克隆人肾与小肠ASBT（顶端Na+/胆汁酸协同转运蛋白）基因并比较2者的序列差别，明确ASBT蛋白在肾小管上皮的亚细胞定位及在人肾组织中的表达情况。方法:从人肾和小肠组织中提取总RNA，然后用带有8肽FLAG标签的PCR引物通过RT-PCR技术扩增ASBT全长cDNA基因并测序，并将其插入真核表达载体中构建ASBT蛋白真核表达载体，然后将其转染到肾小管上皮细胞LLC-PK1中表达并用免疫荧光-激光共聚焦显微镜观察该蛋白的亚细胞定位情况。用免疫组化技术观察ASBT在人肾组织中的表达分布。结果:序列分析结果表明肾小管ASBT基因的序列与小肠ASBT序列完全一致。Western blotting表明ASBT基因在LLC-PK1细胞中得到了正确的表达。共聚焦显微镜分析显示正常ASBT蛋白主要定位于肾小管上皮细胞膜上，与生物信息学的预测结果一致。免疫组化染色表明ASBT蛋白主要表达于人近端肾小管上皮的刷状缘侧，在间质及远端小管没有表达。结论:人肾小管ASBT基因序列与小肠ASBT相同，ASBT蛋白主要表达于近端肾小管上皮细胞管腔侧细胞膜。     

胎盘生长因子及受体在妊娠肝内胆汁淤积症患者胎盘中的表达及其意义

目的探讨胎盘生长因子(PLGF)及受体(F lt-1)在妊娠肝内胆汁淤积症患者胎盘中的表达及其意义。方法采用免疫组织化学方法测定30例妊娠肝内胆汁淤积症患者(ICP组)和30例正常晚期妊娠妇女(对照组)的胎盘组织及8例早孕绒毛组织中PLGF及F lt-1的表达。同时还用抗FⅧRA抗体标记ICP组和对照组孕妇胎盘中的血管密度。结果PLGF及F lt-1在早孕绒毛中主要分布于滋养叶细胞,在ICP患者和对照组胎盘中分布基本一致,主要分布于合体滋养细胞、细胞滋养细胞、血管内皮细胞和绒毛间质细胞。在ICP胎盘中PLGF、F lt-1表达强度及胎盘血管密度均明显低于对照组(P<0.01)。相关分析显示,胎盘组织中PLGF表达强度与胎盘血管密度呈正相关。结论妊娠肝内胆汁淤积症患者胎盘PLGF及F lt-1表达下降与胎盘滋养叶细胞侵入异常、胎盘血管形成异常、胎盘血管密度降低有关,可能是ICP胎儿妊娠结局不良的原因。     

West Nile virus infection of the placenta

Intrauterine infection of fetuses with West Nile virus (WNV) has been implicated in cases of women infected during pregnancy. Infection of timed-pregnant mice on 5.5, 7.5, and 9.5 days post-coitus (dpc) resulted in fetal infection. Infection of dams on 11.5 and 14.5 dpc resulted in little and no fetal infection, respectively. Pre-implantation embryos in culture were also infected with WNV after the blastocyst stage and the formation of trophectoderm. Green fluorescent protein (GFP) expression was observed in a trophoblast stem (TS) cell line after infection with a GFP-expressing WNV construct. However, no fluorescence was observed in differentiated trophoblast giant cell (TGC) cultures. GFP fluorescence was present in TGC cultures if infected TS cells were induced to differentiate. These results suggest that embryos are susceptible to WNV infection after the formation of the trophectoderm around 3.5 dpc through the formation of the functional placenta around 10.5 dpc.     

An IgG-transporting Fc receptor expressed in the syncytiotrophoblast of human placenta

During normal human pregnancy, maternal IgG crosses the placenta and provides passive immunity for the fetus. In so doing, IgG passes through two cellular barriers: the syncytiotrophoblast and the fetal capillary endothelium. The Fc region of IgG is required for its transport across the placenta, but the Fc receptors responsible have not been identified definitively. We recently reported the isolation from a placental cDNA library of clones encoding the α chain of a human homologue of the major histocompatibility complex class I-related Fc receptor, the neonatal Fc receptor (FcRn). In mice, FcRn is essential for the transport of maternal IgG to the fetus and the neonate. We report here the localization of human FcRn mRNA within the placenta by in situ hybridization, and of human FcRn protein by immunohistochemistry. Both methods show that human FcRn is expressed in syncytiotrophoblast, and is, thus, appropriately located to transport maternal IgG across the first barrier. We confirm previous findings that specific binding of IgG to placental membranes is greater at pH 6.0 than pH 7.5. This corresponds with the pH dependence of IgG binding to FcRn and is consistent with the presence of FcRn in syncytiotrophoblast. We propose a transport model in which maternal IgG binds FcRn at low pH in endosomes within the syncytiotrophoblast. FcRn is not expressed in fetal capillary endothelia, and the mechanism of IgG transport across the second barrier remains unknown.     

单纯疱疹病毒Ⅱ型多功能蛋白ICP27在Vero细胞中的表达及定位

目的 构建单纯疱疹病毒Ⅱ型(Herpes simplex virus type 2,HSV-2)感染细胞蛋白27(infected-cell protein 27,ICP27)在Vero细胞中的表达载体,并对其表达进行亚细胞定位.方法 用前期构建成功的pCDNA3.0-ICP27及pEGFP-ICP27通过脂质体介导质粒瞬时转染Vero细胞,经Western blot检测ICP27蛋白的表达.并通过荧光显微镜及免疫组化的方法观察ICP27在Vero细胞中的亚细胞定位.结果 ICP27可在Vero细胞中高表达,pEGFP-ICP27转染Vero细胞12h后,就可以在少量的细胞中表达,而在转染后的24h-48h表达量最大.但转染72h后,表达的蛋白含量在细胞中减少.Western blot结果显示,pEGFP-ICP27和pCDNA3.0-ICP27均可表达ICP27,用质粒pEGFP-ICP27转染的Vero细胞,观察荧光却只发现ICP27定位于细胞核中,而用免疫组化方法检测pCDNA3.0-ICP27发现细胞核和细胞质中均有表达.结论 pEGFP-ICP27和pCDNA3.0-ICP27均可成功的使ICP27在Vero细胞中有效表达,且ICP27在细胞核和细胞质中均有表达,但主要表达于细胞核     

Visualization of lectin-like proteins in human placenta by means of anti-plant lectin antibodies

Proteins antigenically cross-reactive with lectins were sought in the placenta by immunohistochemistry using polyclonal antibodies raised in rabbit against four well-known lectins: Concanavalin A, Wheat germ agglutinin, Ulex europaeus agglutinin, and Phaseolus vulgaris leukoagglutinin (PHA-L), as well as one antibody raised in goat against PHA-L. Even at high dilutions of the primary antibody, strong staining was obtained after short incubations, in patterns generally resembling those obtained for placental lectins by other means, such as those based on binding capacity for glycosylated probes. One of the immunohistochemical patterns distinguishes with great clarity between the trophoblast cell layers, thus relating to developmental and functional parameters; another localises PHA-L-immunoreactivity to the syncytiotrophoblast. These results underline the validity of the immunohistochemical screening as an approach in its own right. Both positive and negative controls were applied to the immunohistochemical methodology. These controls showed that the staining patterns obtained relate to the specificities of the primary antibodies employed; i.e. to lectins. The PHA-Llike cross-reactivity was analysed immunochemically. In electrophoretically separated and Western-blotted placental extracts there were found anti-PHA-L-binding fractions of apparent molecular weights 30 kDa, 58 kDa and 67 kDa. Control studies of the PHA-L antigen showed anti-PHA-L-binding fractions of approximate molecular weights 32 kDa and 60 kDa. The 30 kDa fraction from placenta and the 32 kDa fraction from PHA-L antigen bound lactosylated BSA but not fucosylated BSA. Taken together, the immunohistochemical and biochemical data reveal the presence in the placenta of lectins, one of which resembles PHA-L not only antigenically but also in molecular weight and in sugar-binding specificity.Dedicated to Prof. Dr. med. H. Leonhardt on the occasion of this 75th birthday     

Intercellular junctions in the full term human placenta

Summary Inter-and intrasyncytiotrophoblastic junctions within the human full term placenta were electronmicroscopically investigated using thin sections and freeze-fracturing. Narrow clefts were occasionally situated between surface areas where adjacent chorionic villi exhibited close contact. Whithin these clefts, extensive zonulae and maculae occludentes and numerous maculae adherentes were found. The zonulae occludentes showed a continuous and irregular course on the membrane surface, and the maculae occludentes were irregularly distributed over extended membrane areas.Besides these areas, maculae occludentes and maculae adherentes were observed on infoldings and invaginations of the syncytiotrophoblastic surface membrane. Investigations of the inner surface of the syncytiotrophoblastic layer, that is, the layer facing the villous stroma, also revealed invaginations joined by maculae adherentes.The functional significance of the inter-and intrasyncytiotrophoblastic junctions is discussed with respect to the differentiation of the trophoblast.