脂多糖经线粒体途径诱导细胞滋养细胞凋亡

目的:探讨脂多糖(Lipopolysaccharide,LPS)诱导细胞滋养细胞凋亡的机制。方法:从正常早孕绒毛分离细胞滋养细胞,用无血清培养基培养。在培养基中加入不同浓度的LPS,使其终浓度分别为0、25、50、100、200ng/ml。用光镜和透射电镜观察细胞滋养细胞的形态学变化;用Western blot检测Bax、Bcl-2和Caspase-8的表达。结果:LPS作用后24h,光镜和电镜下见细胞滋养细胞出现凋亡的形态学变化;Western blot检测结果表明,LPS抑制Bcl-2表达,促进Bax和Caspase-8表达。结论:LPS能够经线粒体途径诱导细胞滋养细胞凋亡。     

米非司酮用于足月妊娠引产后的胎盘病理学改变

目的 观察米非司酮用于足月妊娠引产后的胎盘病理改变。方法 将２０例引产指征为预防妊娠过期的孕妇随机分为２组,治疗组１０例予米非司酮５０ｍｇ每日２次口服,连服２天,每３天予米索２５μｇ每６小时阴道塞置;对照组予蒂落安２００ｍｇ每日１次静脉椎注,连用３天,第４天人工破膜后,静脉滴注催产素２．５ＩＵ加５％葡萄糖５００ｍｌ,每分钟滴始,两组均于分娩后取胎盘及蜕膜标本在光镜及电镜下观察。结果 与对照组相比,     

细胞凋亡与妊高征的研究

细胞凋亡是一种在基因调控下的细胞主动死亡过程,以形态和生化上出现一定的改变为特征,与机体的生长发育、细胞分化和病理状态有着十分密切的关系,是细胞死亡的一种形式.生理状态下参与正常妊娠孕卵的着床及胚胎的发育,而在病理状态下异常的细胞凋亡又与自然流产、胎膜早破、输卵管妊娠、胎儿宫内发育迟缓等的发病有关.并在妊高征的发病过程中参予了胎盘、脐带血管内皮细胞的脱落、坏死,以及血管的粥样硬化、纤维样坏死和退形性变.     

米非司酮用于足月妊娠引产胎盘电镜观察及定量分析

目的了解小剂量米非司酮(100mg)用于足月妊娠引产时胎盘超微结构的变化.方法对20例米非司酮配伍米索前列醇引产,20例单用米索前列醇引产,20例自然临产,无明显并发症及合并症,经阴道分娩的足月初产妇胎盘进行透射电镜观察并通过图像分析仪对胎盘超微结构进行体视学分析.结果米非司酮组需用米索前列醇的次数较单用米索前列醇组明显减少.合体滋养细胞、细胞滋养细胞、蜕膜细胞、绒毛中毛细血管内皮细胞的超微结构在三组间未发现明显不同@结论足月孕妇口服米非司酮100mg配伍米索前列醇引产对胎盘无明显影响,并未造成胎儿、胎盘缺血缺氧性损害.     

The Role of Apoptosis on Trophoblast Cell Invasion in the Placental Bed of Normotensive and Preeclamptic Pregnancies

Background. Placental development depends on careful coordination of trophoblast proliferation and apoptosis; however, the synchrony of its effect on trophoblast invasion is unknown. Objective. To examine the relationship between trophoblast apoptosis and proliferation in placental bed tissue of preeclamptic and normotensive pregnancies. Methods. Serial sections from archived placental bed biopsies of 12 normotensive (group 1) and 12 preeclamptic (group 2) were immunolabeled with a rabbit anti-Ki67 antibody, a mouse anti-cytokeratin 18 and its neo-epitope, and a monoclonal cytodeath M30 antibody. Results. The immunoexpression of Ki67 for all trophoblast cell subpopulations within the myometrium was non-reactive in both study groups. Smooth muscle cells of the microvasculature reflected a moderate degree of proliferation in both groups. Morphometric image analysis of the wall of the spiral artery revealed a mean area of 31,1729?±?51,180?µm² compared to 35,795?±?8045?µm² in groups 1 and 2, respectively. An elevation of intramural trophoblast was evident within the spiral artery of group 1 (13%). Comparative analyses of M30 distribution on corresponding serial sections were 0.06% versus 0% in groups 1 and 2, respectively. The mean field area percentage of interstitial trophoblast invasion was 10.79% versus 2.87% with corresponding areas of apoptosis been 0.8 % versus 1.9 % in groups 1 and 2, respectively. Conclusions. This study demonstrates an increased trophoblast apoptosis in placental bed of preeclamptic compared to normotensive pregnancies with concurrent absence of proliferation at term.     

妊高征患者血浆和胎盘中环磷酸鸟苷浓度的变化及其意义

目的　探讨环磷酸鸟苷 (c GMP)在妊高征发病中的作用。　方法　采用放射免疫法测定 12 4例正常妊娠各期、32例妊高征患者及 30例健康对照者血浆 c GMP浓度。其中 32例正常足月孕妇和 32例妊高征患者于剖宫产时取胎盘组织测其匀浆 c GMP浓度。　结果　 (1)正常妊娠各期血浆 c GMP(2 6 .2 5± 4.30 )、(2 3.40± 4.5 4)、(2 6 .2 0± 6 .6 0 )、(34 .14± 8.41)、(32 .32± 8.11) nm ol/ L 显著高于健康对照者 (2 0 .2 2± 3.44 ) nm ol/ L (P均 <0 .0 5 )。 (2 )妊高征血浆及胎盘 c GMP(2 8.49± 8.98) ) nmol/ L与 (2 .76± 0 .49) nmol/ g均显著低于正常足月孕妇 (34 .14± 8.41) nmol/ L与(3.78± 0 .81) nmol/ g(P<0 .0 5 ,P<0 .0 0 1)。轻度妊高征患者血浆及胎盘 c GMP显著高于中度及重度患者 (P<0 .0 0 5 ,P<0 .0 5及 P<0 .0 5 ,P<0 .0 0 1)。妊高征患者血浆及胎盘 c GMP与舒张压均呈负相关 (P<0 .0 5 ,P<0 .0 5 )。 (3)妊高征患者胎盘 c GMP显著低于其血浆 c GMP(P<0 .0 0 1)。　结论　妊娠期间胎盘产生的 c GMP可能在妊高征的发病中起重要作用     

雷公藤内酯醇诱导上皮性卵巢癌Caov-3凋亡的实验研究

目的 :观察雷公藤内酯醇对上皮性卵巢癌体外生长特性的影响 ,并进一步探讨有关的信号转导机制。方法 :分别采用MTT法、流式细胞仪测定和克隆形成实验观察雷公藤内酯醇对上皮性卵巢癌Caov 3增殖、凋亡和侵袭能力的影响 ,以Westernblot方法检测相关的信号分子。结果 :雷公藤内酯醇显著地抑制了Caov 3的增殖和克隆形成能力 (P <0 .0 5 ) ,诱导其凋亡 ,caspase途径和ERK活化主要参与了诱导凋亡的过程。结论 :雷公藤内酯醇对上皮性卵巢癌Caov 3生长特性的影响 ,使其有望成为一种新的化疗药物应用于卵巢癌的治疗。     

Increased Placental Apoptosis in Maternal Food Restricted Gestations: Role of the Fas Pathway

The placenta has a pivotal role, shuttling nutrients to the developing fetus and producing hormones essential to pregnancy. Maternal food restriction (MFR) during pregnancy results in growth restricted newborns, a consequence attributed primarily to maternal nutrient supply. We hypothesized that MFR further compromises fetal growth by decreased placental growth or increased placental apoptosis. We determined the potential pathway (Fas–Fas-ligand; Fas-FasL) of placental apoptosis in MFR pregnancies. We assessed the two morphologically and functionally distinct zones (basal and labyrinth) at embryonic age 20 (E20) in ad libitum fed controls (AdLib) and 50% MFR placentas. We studied fetuses and placentas from both proximal and mid-horn positions to evaluate any differential impact by MFR. Placenta apoptosis was quantified using terminal dUTP nick-end labeling (TUNEL) assay and the data were compared to immunodetection of cleaved caspase-3, Fas and FasL followed by Western blot quantification of Fas, FasL, caspase-8 and -3, tBID, and poly-ADP-ribose polymerase (PARP). MFR reduced maternal, fetal and placental basal and labyrinth weights. The results suggest that the increased apoptosis may be mediated, in part, via Fas pathway and the defective placental development in the MFR may be a major contributor to the decreased fetal growth.     

重度妊娠高血压综合征患者胎盘组织细胞凋亡及其调控基因 …

目的　研究细胞凋亡及其调控基因在重度妊娠高血压综合征 (妊高征 )发生、发展中的作用。方法　对 40例正常晚孕妇女 (对照组 )和 40例重度妊高征患者 (观察组 )的胎盘组织进行分析。用DNA缺口原位末端标记 (TUNEL)技术检测细胞凋亡 ;免疫组织化学方法检测促进和抑制凋亡基因 (bax/bcl 2 )的表达。结果　对照组胎盘细胞滋养细胞、合体滋养细胞中凋亡指数分别为 ( 1.1±0 .9) %、( 41.8± 1.5 ) % ;bax阳性率分别为 ( 1.0± 0 .9) %、( 2 8.9± 9.7) % ;bcl 2阳性率分别为 ( 2 .2±0 .8) %、( 2 2 .9± 0 .7) % ,总bax/bcl 2为 0 .7～ 1.7。观察组胎盘细胞滋养细胞、合体滋养细胞的凋亡指数分别为 ( 4.3± 1.2 ) %、( 45 .3± 1.4) % ;bax阳性率分别是 ( 2 .2± 0 .8) %、( 42 .5± 11.7) % ;bcl 2阳性率分别是 ( 3 .2± 0 .8) %、( 2 3 .3± 7.8) % ;总bax/bcl 2为 1.0～ 3.2。即 :对照组胎盘中有一定量的细胞凋亡、bax、bcl 2表达 ,bax/bcl 2表达间呈平衡趋势 ;观察组中细胞凋亡明显增高 (P <0 .0 1) ,bax表达也明显增强 (P <0 .0 1) ,bcl 2表达仅呈增高趋势 ,但差异无显著性 (P >0 .0 5 ) ,bax/bcl 2比值明显增高(P <0 .0 1)。结论　细胞凋亡及其调节基因的表达间具有一致性 ;细胞凋亡、bax/bcl 2表达     

复发性流产患者胎盘滋养细胞凋亡及Fas、FasL表达的研究

目的 :通过比较正常早孕妇女和复发性流产 (RSA)患者胎盘绒毛滋养细胞凋亡及Fas、FasL表达 ,进一步从细胞及分子水平探讨复发性流产的发病机理。方法 :应用免疫组织化学SP方法 ,测定 30例RSA患者和 2 5例正常早孕妇女胎盘绒毛滋养细胞Fas和FasL基因蛋白的表达 ,并通过MIAS 2 0 0 0医用彩色病理图像免疫组化测量系统对其表达进行半定量分析 ,其结果用平均灰度表示 ;同时应用DNA缺口原位末端标记技术 (TUNEL法 )测定两组胎盘绒毛滋养细胞凋亡情况。结果 :RSA组患者胎盘绒毛FasL的表达低于正常组 ,差异有显著性 (P <0 0 5 ) ;而Fas表达两组比较差异无显著性 (P >0 0 5 ) ,胎盘绒毛细胞滋养细胞及合体滋养细胞的凋亡指数 ,RSA组明显高于正常组 ,差异有非常显著性 (P<0 0 1)。结论 :细胞凋亡的增加可能是RSA发生的一个重要病理过程 ,且RSA患者胎盘绒毛滋养细胞表面FasL表达减少 ,导致母胎间免疫耐受的破坏 ,引起异常的免疫反应可能是RSA发病的重要机制。     

Characterization of Uterine Contractures and Demonstration of Placental Vasodilation in the Mid-Gestation Albino Guinea Pig

Twenty-five mid-gestation guinea pigs were studied to determine the nature of spontaneous uterine contractures and elucidate the temporal relationship between blood flow and contractile activity. In the first protocol, 15 mid-gestation guinea pigs with indwelling vascular catheters were studied. Blood flows were measured by radioactive microsphere technique during rest, uterine contracture, and, in six animals, 1 minute after contracture activity. During contracture, maternal blood flow to the placenta closest to the electrode fell to 81.3 ± 5.3% of control (n = 15, P ±. 01) and then rose to 130.4 ± 10.2% of control (P ≤. 01) 1 minute after a contracture. The corresponding relationships were also significant for placental vascular resistance and constitute a reactive hyperemic response of the placenta to uterine contracture. No such phenomenon was observed in the more remote placentas on either the ipsilateral or contralateral side.

To investigate the nature of contractile activity, electrodes were placed for measurement of uterine electromyogram in ten mid-gestation guinea pigs. Uterine activity was studied in conscious guinea pigs on the day of surgery and 2 days later. Time spent in synchronous activity fell from 72.9 ± 9.8 on the day of surgery to 24-4 ± 8.4% (P ≤. 01) 2 days later; asynchronous contractile activity rose from 13.9 ± 5.9 to 44.6 ± 6.4% (P ≤. 01), respectively. We conclude that in the guinea pig, contractures are often a local phenomenon during mid-gestation and that they exert an influence on local placental blood flows both during and immediately after contracture.     

Characterization of the Placental Macrophage Secretome: Implications for Antiviral Activity

It is well documented that placental macrophages show lower levels of HIV-1 infection than monocyte-derived macrophages (MDM). We used proteomic methods to test the hypothesis that placental macrophages secrete different proteins as compared to MDM that may contribute to decreased HIV-1 replication. Placental macrophages and MDM were cultured for 12 days and supernatant was collected. To characterize supernatants, the protein profiles of placental macrophages and MDM were compared using the protein chip assay. Subsequently, proteins were separated by one-dimensional gel electrophoresis and identified by tandem mass spectrometry at the corresponding mass to charge (m/z) range of 5000–20,000. Significant differences were found between placental macrophages and MDM in seven protein peaks with m/z values of 6075, 6227, 11,662, 14,547, 6158, 7740, and 11,934 on the CM10 and IMAC chips. After sequencing and identification, five proteins were validated for differential expression in placental macrophages and MDM by Western blot analyses. Peroxiredoxin 5, found to be more abundant in placental macrophage supernatants, is important in the cellular antioxidant mechanisms, and other members of its family have shown antiviral activity. Cystatin B was less abundant in PM supernatant, and decreased intracellular levels have recently been shown to be associated with lower HIV-1 replication in placental macrophages than in MDM. This study elucidates for the first time the placental macrophage secretome corresponding to 5000–20,000 Da and advances our understanding of the proteins secreted in the placenta that can protect the fetus against HIV-1 and other viral infections.     

拓扑替康诱导人卵巢癌顺铂耐药细胞株COC1/DDP凋亡的研究

目的探讨拓扑替康(TPT)诱导人卵巢癌顺铂耐药株COC1/DDP凋亡的内在分子机制。方法采用透射电镜和TUNEL法观察TPT诱导COC1/DDP细胞凋亡的情况;采用RTPCR和Westernblot检测TPT对COC1/DDP细胞内bcl2、bax和caspase3基因表达的影响以及caspase3活性的改变。结果COC1/DDP细胞在TPT作用后出现典型的凋亡形态特征,且凋亡率由8.54%上升为19.31%(P<0.05);TPT不影响COC1/DDP细胞内bcl2mRNA表达,却明显增加baxmRNA和caspase3mRNA表达,并提高caspase3活性。结论TPT诱导COC1/DDP细胞凋亡可能依赖于细胞内bax蛋白增高和caspase3的活化。     

The Phenotype in Placental Trisomy 7

Trisomy 7, in mosaic state, was identified at chorionic villus sampling. The pregnancy was closely followed, and proceeded uneventfully. Mosaic trisomy 7 was confirmed in the term placenta, the organ having no structural abnormalities; the karyotype of the phenotypically normal baby was 46,XY. Trisomy 7, mosaic or nonmosaic, detected at chorionic villus sampling in an ultrasonographically normal pregnancy, appears typically to be associated with a normal fetal karyotype, and placental growth, structure, and function are not discernibly compromised.     

Isradipine Reduces Blood Pressure,But Not Placental Blood Flow in Pregnancy Induced Hypertension

Thirteen pregnant women with pregnancy induced hypertension (PIH) in the third trimester, and eleven normotensive women in the third trimester were given 0.3 mg of the novel dihydropyridine calcium antagonist isradipine intravenously (iv). After drug administration, maternal systolic and diastolic blood pressure (SBP and DBP) decreased in both PIH patients and controls. In the hypertensive patients SBP was reduced from a mean of 150 to 124 mmHg (p<0.001), and in the controls from 124 to 119 mmHg (p>0.5). Corresponding figures for DBP were 99 to 83 mmHg (p<0.001) and 79 to 70 mmHg (p<0.001) respectively. The maximum effects were seen within 15 minutes. Maternal heart rate (MHR) increased significantly, in the PIH group from 79 to 93 beats per minute (bpm), and in the normotensive group from 84 to 98 bpm, respectively. Maximum effects were observed within 15 minutes, and within 90 minutes MHR returned to initial values. There was a non significant, transient increase in fetal heart rate in both groups after isradipine. Blood flow in the umbilical and arcuate arteries, expressed as the pulsatility index and the resistance index, was not significantly affected after 10 and 30 minutes in any of the women. Few, mild side effects (headache, palpitations) were observed. The results suggest that isradipine given iv, effectively lowers BP in normal pregnant subjects as well as in PIH patients without affecting uteroplacental blood flow.     

输卵管积液对小鼠胚胎体外发育的影响

目的 :了解输卵管积液是否具有胚治毒性。方法 :收集兔输卵管液和机械诱导的输卵管积液 ;小鼠超数排卵 ,收集 2 -细胞胚胎 ,随机分配到含有不同浓度输卵管积液、输卵管液 (正常对照 )和培养液 (空白对照 )中培养 ,观察并记录胚胎形态 ,同时测定新形成囊胚的细胞数和有丝分裂指数。结果 :1 0 0 %输卵管积液组囊胚形成率和胚胎孵化率均明显低于两对照组 (P<0 .0 0 1 ) ;而低浓度 (2 0 %和 1 0 % )积液组囊胚形成率和胚胎孵化率与两对照组相比 ,均无明显差异。各实验组囊胚的细胞数和有丝分裂指数均明显低于两对照组(P<0 .0 5)。结论 :机械诱导的输卵管积液 ,会影响早期种植前胚胎质量