Fas-Fas Ligand System-Induced Apoptosis in Human Placenta and Gestational Trophoblastic Disease

PROBLEM: Previous studies demonstrated that unique tissue-specific antigens expressed on vascular endothelial cells can serve as immunogens, and the apparent association between transplant rejection and antiendothelial cell (EC) antibodies is well established. A common feature of some placentas from women with recurrent pregnancy loss is the diffuse formation of microthrombi associated with changes in thromboresistant properties of endothelial cells, similar to the findings in rejected organs. Therefore, the prevalence of anti-EC antibodies in patients with recurrent pregnancy loss and the role of these antibodies in cultured human endothelial cells from umbilical cord vein were studied. METHOD OF STUDY: To evaluate the frequency of anti-EC antibodies, sera from 160 nonpregnant patients with recurrent pregnancy loss after absorption with pooled platelets and pooled leukocytes were tested using cultured human umbilical cord vein endothelial cells (HUVEC) by flow cytometry. To study the role of anti-EC antibodies, purified anti-EC IgGs were added to HUVEC cultures and hemostatic, fibrinolytic, and anticoagulation pathway markers (tissue factor, tissue plasminogen activator, palsminogen-activator inhibitor, thrombomodulin, heparan-sulfate proteoglican, antithrombin III, von Willebrand factor, CD54, human leukocyte antigens-DR, and transferin receptor) were detected by specific antibodies and flow cytometry. Immunoblotting analyses were done by using purified anti-EC IgGs against cell membrane proteins from endothelial cells and leukocytes extracted by detergent solubilization. RESULTS: Thirty-nine (24%) of the patients were positive for EC antibodies. Antipaternal lymphocyte antibodies were tested as well and were found in 37 (23%) of the patients as 25 sera (15.6%) showed reactivity with both EC and lymphocytes. Certain patient sera were reactive with HUVEC lines from some but not other umbilical cords, which suggests allotype Sera from 70 normal healthy male and nonpregnant female controls did not react with any of the individual HUVEC lines used. The purified anti-EC immunoglobulin G (IgGs) recognized bands, from HUVEC surface membrane protein preparations, with molecular weights of 120 kDa. Both hemostatic and fibrinolytic pathway markers were found activated in the presence of anti-EC IgGs, suggesting an altered endothelial cell surface activation state. CONCLUSIONS: The results indicate that anti-EC antibody is another marker for a subset of recurrent spontaneous aborters who may have activation of hemostasis and fibrinolysis as a mechanism involved in their losses.     

彩色多普勒血流显像与彩色多普勒能量图在恶性滋养细胞肿瘤的应用

目的探讨彩色多普勒血流显像(CDFI)及彩色多普勒能量图(CDE)在恶性滋养细胞肿瘤(GTD)早期诊断及化疗监测中的应用价值.方法对20例GTD进行CDFI、CDE检测,观察其血流情况及用PD检测病变区血流速度,测算RI值.结果CDFI显示病变区血流异常丰富,CDE显示更丰富的血流信号.PD检测出低阻血流,RI=0.41±0.07,化疗前后相比CDFI、CDE和PD改变明显,RI值差异有非常显著性意义(p<0.01).结论CDFI、CDE对恶性滋养细胞肿瘤诊断及观察疗效均有十分重要的价值.     

彩色多普勒血流显像与彩色多普勒能量图在恶性滋养细胞肿瘤的应用

目的探讨彩色多普勒血流显像(CDFI)及彩色多普勒能量图(CDE)在恶性滋养细胞肿瘤(GTD)早期诊断及化疗监测中的应用价值. 方法对20例GTD进行CDFI、CDE检测,观察其血流情况及用PD检测病变区血流速度,测算RI值. 结果 CDFI显示病变区血流异常丰富,CDE显示更丰富的血流信号.PD检测出低阻血流,RI=0.41±0.07,化疗前后相比CDFI、CDE和PD改变明显,RI值差异有非常显著性意义(p<0.01). 结论 CDFI、C DE对恶性滋养细胞肿瘤诊断及观察疗效均有十分重要的价值.     

p21-Activated Kinase-1 Promotes Aggressive Phenotype,Cell Proliferation,and Invasion in Gestational Trophoblastic Disease

Gestational trophoblastic disease (GTD) includes hydatidiform mole (HM), which can develop persistent gestational trophoblastic neoplasia requiring chemotherapy; choriocarcinoma, which is a frankly malignant tumor; placental site trophoblastic tumor; and epithelioid trophoblastic tumor. p21-Activated kinases (PAKs) promote malignant tumor progression. Therefore, this study investigated PAK1, PAK2, and p-PAK2 Ser²⁰ in the pathogenesis of GTD. By real-time PCR, PAK1 mRNA was significantly higher in HMs, particularly metastatic HMs (P = 0.046) and HMs that developed persistent disease (P = 0.011), when compared with normal placentas. By immunohistochemistry, significantly increased cytoplasmic PAK1 immunoreactivity in cytotrophoblasts was also detected in HMs (P = 0.042) and choriocarcinomas (P = 0.003). In addition, HMs that developed persistent disease displayed higher PAK1 immunoreactivity than those that regressed (P = 0.016), and elevated PAK1 immunoreactivity was observed in placental site trophoblastic tumors. Indeed, there was significant positive correlation between PAK1 expression and the proliferative indices Ki-67 (P = 0.016) and MCM7 (P = 0.026). Moreover, higher PAK1 mRNA and protein expression was confirmed in the choriocarcinoma cell-lines JEG-3 and JAR; however, PAK2 mRNA and p-PAK2 immunoreactivity showed a similar expression pattern in normal first trimester placentas and GTD. Knockdown of PAK1 in JEG-3 and JAR reduced cell proliferation, migration, and invasion ability, up-regulated p16, and down-regulated vascular endothelial growth factor and MT1-MMP expression. This is the first report revealing the involvement of PAK1 in the pathogenesis and clinical progress of GTD.Gestational trophoblastic disease (GTD) is a disease of the placenta resulting from abnormal trophoblastic proliferation. It encompasses hydatidiform mole (HM), choriocarcinoma (CCA), placental site trophoblastic tumor (PSTT), and epithelioid trophoblastic tumor. The latter three are malignant tumors, whereas HMs are nonneoplastic lesions with an increased risk of malignant transformation. Furthermore, 8% to 30% of HMs may persist after suction evacuation and develop gestational trophoblastic neoplasia (GTN), hence requiring chemotherapy. Although the response of GTD to chemotherapy is in general very good, chemoresistant cases still exist despite advances in chemotherapy. While there is growing understanding in the molecular biology of GTD, the precise molecular pathways of its development should be further explored.^1,2,3p21-Activated kinases (PAKs) were initially discovered as effector molecules of Rho GTPases, Rac, and CDC42,⁴ and are increasingly recognized as important mediators for a wide variety of cellular functions, including cell morphogenesis, motility, mitosis, apoptosis, and angiogenesis.^5,6 PAKs have been divided into group I (PAKs1-3) and group II (PAKs4-6), bases on structural organization and mode of regulation.⁵ In group I PAKs, the p21-binding domain in the N-terminal domain binds small GTPases Rac and CDC42 to cause autophosphorylation of certain sites in the N-terminal inhibitory domain, leading to conformational change that releases the auto-inhibition and activates the kinase activity.⁵ Phosphorylation of PAK1 at Ser²¹ regulates the binding of PAK1 to Nck, resulting in PAK1 cycling between membrane and cytosol and finally facilitates cell migration.^7,8 Its corresponding phosphorylation site at PAK2 is Ser²⁰.Overexpression of PAK1 has been observed in various human cancers,⁶ such as breast, colorectal, and ovarian cancers.⁹ PAK1 was found to regulate the invasiveness of breast cancer cells and the malignant progression of colorectal carcinomas to metastasis.^10,11 PAK2 has also been found to be involved in Rac3-mediated breast cancer cell proliferation.¹² The recent development of kinase inhibitors for the PAK family molecules^13,14 is intriguing since they may be explored as potential therapeutic targets in chemoresistant GTD.In this study, we investigated the expression and localization of PAK1, PAK2, and p-PAK2 Ser²⁰ in normal human placenta and in GTD to determine whether PAK1 or PAK2 are involved in development and malignant progression of GTD. We also correlated their expression with clinical outcome and examined the in vitro effects and potential downstream targets of PAK1 on CCA cells.     

Detection of Early Pregnancy Factor-Like Activity in Women With Gestational Trophoblastic Tumors

ABSTRACT: The presence of immunosuppressive early pregnancy factor (EPF) in the maternal serum has so far been associated with gestation. Its presence in the serum of women with gestational trophoblastic tumors was investigated. The results indicate that while EPF activity was detected in the serum of women with choriocarcinoma, no such activity was detected in the serum of women with hydatidiform mole, leading to the novel use of EPF as a marker to distinguish these two clinical situations. Results of the experiments also suggest that EPF moiety present in the maternal serum during pregnancy may be of different molecular entity than that present in the serum of women with choriocarcinoma.     

The HL-A and ABO Antigens in Trophoblastic Disease

No statistically significant deviations in phenotype frequencies of the 25 HL-A antigens or the ABO antigens were seen when 111 Caucasian patients with trophoblastic disease were compared with 1,259 healthy Caucasian controls. However, an increasein the incidence of HL-A11 was found in 39 patients who currently had the disease, but not in 72 who had recovered from the disease. Further, an increase in the frequency of W18 was observed among 18 patients who currently had 'invasive' disease (choriocarcinoma or invasive mole), but not in 44 who had recovered from such disease. If valid for larger patient population, these increases may suggest association of HL-A11 and W18 with the 'morbidity' of the trophoblastic disease. No increase in histocompatibility was seen in 45 patient-couples over 67 control-couples in terms of decrease in the number of male spouse's HL-A incompatibilities, and no significant difference was seen in the distribution of pregnancies in the two groups. No significant difference was observed in the incidence of different male-female combinations of ABO blood groups between 95 patient-couples and an equal number of control-couples. Lymphocytotoxic antibodies were found in 64 patients (158 sera) or 36% of the 178 patients (413 sera) examined. HL-A specific antibodies were found in 30 or 17% patients (39 sera). Of these 30, 24 patients had molar pregnancies and six had choriocarcinoma. Whether these antibodies have a role in the destruction of neoplastic tissue remains to be determined.     

HLA Antigen Sharing in Italian Couples in Which Women Were Affected by Gestational Trophoblastic Tumors

PROBLEM: The development of gestational trophoblastic tumors (GTT), in which genetic factors are strongly involved, is a rare event. To test the possibility that gene(s) linked to the Major histocompatibility Complex (MHC) may have a role in both embryo growth and tumor development, the HLA typing was performed on patients affected by GTT and on their partners. METHOD: The study group of sixteen couples, in which the women were affected by an invasive mole or choriocarcinoma, and the control group of thirty normal fertile couples without history of spontaneous abortion or GTT were typed for class I and class II HLA antigen. RESULTS: The results showed no differences in single HLA-A and B antigen frequency between GTT couples and controls. In HLA-DR, locus an increased frequency of DR-6 antigen was observed (p<0.05). No differences were observed in the frequency of number of antigens shared. When considering the single locus no differences were found in the sharing of the antigens of the A and B locus, while the frequency of antigenic sharing for DR locus was significantly higher in GTT couples with respect to controls (p<0.025). Furthermore a higher frequency of Bw35-DR5 antigenic combination was found in GTT partners than in controls (P<0.02). CONCLUSIONS: These data represent a confirmation of the existence of a MHC linked gene(s) influencing the GTT development.     

1例滋养细胞疾病患者化疗后并发严重口腔溃疡护理

滋养细胞疾病患者早期即可发生血行转移,单穿手术达不到根治的目的,所以要以化疗为主。口腔溃疡是患者在化疗过程中常发生的一种副反应,它不仅给患者造成痛苦,且影响患者营养供给和化疗的顺利进行,而且也是患者感染、发热并发败血症的主要原因。口腔溃疡的发生率及严重程度与口腔护理直接相关。本文主要回顾分析在我院进行滋养细胞化疗的1例患者的有关资料,探讨其发生口腔溃疡的原因,并提出相应的护理对策。     

HL-A Matings in Trophoblastic Neoplasia

Seventy-five patients with trophoblastic neoplasia and their husbands were typed for the antigens of the HL-A system. The phenotypic mating frequencies were analysed separately for both subloci. The expected number of matings for each antigenic combination was calculated using the reaction frequencies of antigens actually observed in the patients and their husbands. The analysis showed that, as far as these antigens were concerned, mating was at random. The risk of a woman getting a trophoblastic neoplasm is apparently not influenced by her choice of mate as far as the HL-A system is concerned.     

Gestational surrogacy

Gestational surrogacy is a treatment option available to women with certain clearly defined medical problems, usually an absent uterus, to help them have their own genetic children. IVF allows the creation of embryos from the gametes of the commissioning couple and subsequent transfer of these embryos to the uterus of a surrogate host. The indications for treatment include absent uterus, recurrent miscarriage, repeated failure of IVF and certain medical conditions. Treatment by gestational surrogacy is straightforward and follows routine IVF procedures for the commissioning mother, with the transfer of fresh or frozen-thawed embryos to the surrogate host. The results of treatment are good, as would be expected from the transfer of embryos derived from young women and transferred to fit, fertile women who are also young. Clinical pregnancy rates achieved in large series are up to 40% per transfer and series have reported 60% of hosts achieving live births. The majority of ethical or legal problems that have arisen out of surrogacy have been from natural or partial surrogacy arrangements. The experience of gestational surrogacy has been largely complication-free and early results of the follow-up of children, commissioning couples and surrogates are reassuring. In conclusion, gestational surrogacy arrangements are carried out in a few European countries and in the USA. The results of treatment are satisfactory and the incidence of major ethical or legal complications has been limited. IVF surrogacy is therefore a successful treatment for a small group of women who would otherwise not be able to have their own genetic children.     

Trophoblastic neoplasia in an African urban population.

A clinical study of trophoblastic neoplasia in a Nigerian population in Lagos over a four-year period is reported. A high incidence of one in 379 deliveries for benign trophoblastic tumor and one in 846 deliveries for malignant tumor was found. Seventeen percent of benign trophoblastic tumors in this series progressed to the malignant type, but malignant trophoblastic tumor was preceded by the benign type (hydatidiform mole) in 46 percent of cases. The anterior vaginal wall is a common site for metastases of malignant trophoblastic neoplasia and, in one patient, the lesion progressed further to form a vesicovaginal fistula. While the management of benign disease was conservative, all cases of malignant trophoblastic neoplasia received chemotherapy.     

Gestational diabetes mellitus

Gestational diabetes (GDM) is defined as any degree of glucose intolerance with onset or first recognition during pregnancy and is associated with increased feto-maternal morbidity as well as long-term complications in mothers and offspring. GDM is diagnosed by an oral glucose tolerance test (OGTT) or fasting glucose concentrations in the diabetic range. In case of a high risk for GDM/type 2 diabetes (history of GDM or prediabetes [impaired fasting glucose or impaired glucose tolerance]; malformation, stillbirth, successive abortions or birth-weight > 4500 g in previous pregnancies) performance of the OGTT (120 min; 75 g glucose) is recommended already in the first trimester and--if normal--the OGTT should be repeated in the second/third trimester. In case of clinical symptoms of diabetes (glucosuria, macrosomia) the test has to be performed immediately. All other women should undergo a diagnostic test between 24 and 28 gestational weeks. If fasting plasma glucose exceeds 95 mg/dl, 1 h 180 mg/dl and 2 hrs 155 mg/dl after glucose loading (OGTT) the woman is classified as GDM (one pathological value is sufficient). In this case a strict metabolic control is mandatory. All women should receive nutritional counseling and be instructed in blood glucose self-monitoring. If blood glucose levels cannot be maintained in the normal range (fasting < 95 mg/dl and 1 h after meals < 130 mg/dl) insulin therapy should be initiated. Maternal and fetal monitoring is required in order to minimize maternal and fetal/neonatal morbidity and perinatal mortality. After delivery all women with GDM have to be reevaluated as to their glucose tolerance by a 75 g OGTT (WHO criteria).     

Review: Analogies Between Trophoblastic and Malignant Cells

PROBLEM: The question of how trophoblastic and malignant cells evade immunologic recognition and rejection by their host was studied. METHOD OF STUDY: A literature review was conducted. RESULTS: Trophoblastic and malignant cells share a number of similarities. These include a lack of major histocompatibility complex antigen expression, resistance to lysis by natural killer cells, T-helper cell-2 (TH2)-biased response, prostaglandin E production, and response to transforming growth factor beta. In addition, the analogies between trophoblastic and malignant cells extend into immunotherapy in which anti-idiotype therapy has a viable role in the prevention of pregnancy loss and the treatment of cancer. CONCLUSIONS: Trophoblastic and malignant cells use a number of similar mechanisms to resist rejection by their host. By using similar strategies these cells are able to successfully co-exist in an immunologically hostile environment.     

Trophoblastic tumor at the site of the placenta

The author describes the histological structure of a rare placental site trophoblastic tumor. In differential diagnosis it is necessary to take into account hydatidiform mole and chorionepithelioma. In differentiating with hydatidiform mole lack of placental villi is characteristic while with choriocarcinoma the presence of connective stroma in PSTT is helpful.