Monitoring Early Production of Chorionic Gonadotrophin (HCG) Following In Vitro Fertilization and Embryo Transfer

Summary: A rapid, sensitive assay was developed for monitoring HCG production and applied to serum and urine collection between days 4 and 20 after OPU in 55 patients treated by ovarian hyperstimulation and IVF. Eleven ongoing pregnancies, 2 ectopic pregnancies and I clinical abortion resulted. The earliest detection of a rise in HCG was on day 7 after OPU. The HCG values showed that trophoblastic tissue functioned transiently in another 11 patients and probably in another 5. Thus evidence for functioning trophoblastic tissue was obtained in a total of 30 patients (55%). These findings are important for the further improvement of IVF. No antibodies to administered HCG were detected.     

Determination of the concentration of chorionic gonadotropin (HCG) and its free subunits in placental tissue during various periods of normal pregnancy

The aim of this study was to estimate HCG and its free subunits (alpha and beta) content in placental tissue and to determine the characteristic pattern of their ratio changes in the course of normal pregnancy. For this we used 47 samples of placental extracts. The tissue was extracted and gel filtrated on a calibrated Sephadex-G-100 column. Crude placental extracts and each fraction after chromatography were radioimmunoassayed for HCG, alpha HCG and beta HCG in a double antibody, homologous radioimmunoassay system. We obtained the following results: Peak of native HCG secretion occurred in 6-8 gestational week thereafter its concentration in placenta decreases. The absolute amount of free alpha HCG increased after the first trimester like to native HCG, but the relative quantity of alpha HCG exceeded that for HCG. In extracts from mature placenta quantity of free alpha HCG was about 5-10-fold more than HCG. In chorionic extracts from the first trimester we have proved the free beta subunit of HCG. It has not been found in extracts from mature placenta.     

Potential control of fertility in women by immunization with HCG

Since HCG plays a role in maintaining pregnancy, methods of interfering with its secretion offer promise in fertility control. HCG antibodies raised in laboratory animals have been shown to neutralize the effects of exogenously administered HCG. Immunization of humans with HCG will not result in antibody production since it is chemically similar to pituitary LH, but immunization with altered HCG has produced antibodies reacting to HCG and LH. Because of this dual neutralization, no human application is possible without the risk of disrupting ovarian function in women of productive age. Studies have been carried out in which women were immunized with conjugates of the beta subunit of HCG and protein, producing antibodies reacting with native HCG; long-term evaluation of these subjects is awaited. It may be possible to use peptides from the 30C-terminal amino acids of the beta subunit of HCG which have no counterpart in the beta subunit of human LH, but enzymatic digestion to obtain these peptides is costly and time consuming. Preliminary data suggest that immunizations with synthetic HCG peptides conjugated to protein carriers will produce antibodies reacting with natural hormones in vitro, but additional testing is necessary. It is noted that the parameters of antibody specificity, tissue damage, immune complex disease, reversibility, and individual variations in responses must be assessed before the expectations of human use can be suggested.     

Cellular localization of alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG), and carcinoembryonic antigen (CEA) in malignant germ cell tumors of the ovary using immunoperoxidase technique (author's transl)

An immunohistologic study of germ cell tumors of the ovary was undertaken to investigate a correlation of the histology with the presence of alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG), and carcinoembryonic antigen (CEA) in the tissue specimens. A positive reaction for AFP was consisted of dark brownblack granular intracytoplasmic deposits and some of hyaline globules were also positive in some instances. There was tissue localization of AFP in 9 of 10 embryonal carcinomas (endodermal sinus tumor, Teilum) and in 4 of 5 solid teratomas. Two patients with embryonal carcinoma positive for AFP had normal serum level in AFP. A positive staining for HCG was found in 4 embryonal carcinomas and in 2 solid teratomas; it was consisted of dark brown cytoplasmic deposits localized exclusively to syncytiotrophoblast giant cells. The tumor cells positive for AFP but negative for HCG, and vice vasa indicated that these two carcinoembryonic proteins were produced by different cells. The localization of CEA was also studied in 3 embryonal carcinomas. None of embryonal carcinomas and 3 solid teratomas were reacted to CEA; it was consisted of a dark brown deposit localized in cell surface of stratified squamous cells and tall columnar cells forming glandular structure. All of the histologically immature neural components were negative for these carcinoembryonic proteins.     

Formation of HCG in the human fetoplacental unit.

HCG and beta-HCG were determined in umbilical arterial and venous blood immediately after spontaneous term delivery. HCG and beta-HCG are present in higher concentrations in arterial than in venous umbilical cord blood. The data are consistent with the hypothesis that HCG and beta-HCG are produced in the fetal compartment of the human fetoplacental unit, with a larger contribution of the female fetus as compared with the male.     

环状RNA在子宫内膜癌中的表达及临床意义分析

Objective?To investigate the role and mechanism of lncRNA HCG22 in proliferation, migration and invasion of cervical cancer. Methods?The expressions of HCG22 and miR-629-5p in cervical tissues and cells were detected by RT-qPCR. HeLa cell were allocated into pcDNA3.1 group, pcDNA3.1-HCG22 group, and pcDNA3.1-HCG22+miR-629-5p mimics group. The HeLa cell growth, migration and invasion were assessed by MTT, wound-healing and Transwell assays, respectively. The interaction relationship between HCG22/miR-629-5p and miR-629-5p/FOXO3 were analyzed by luciferase reporter. Results?HCG22 was significantly decreased in cervical cancer tissues and cell lines compared with paired paracancerous tissues or END1/E6E7 cell (P<0.01). The upregulation of HCG22 significantly inhibited tumor cell proliferation (P<0.01), migration (P<0.01), and invasion (P<0.01) compared with pcDNA3.1 group. miR-629-5p expression was increased in cervical cancer tissues and cell lines compared with paired paracancerous tissues or END1/E6E7 cell (P<0.01). There was a negative correlation between HCG22 and miR-629-5p (r=-0.7661, P<0.01). The addition of miR-629-5p mimics reversed the suppressive effects of pcDNA3.1-HCG22 on cell proliferation, migration, and invasion (P<0.01). Both HCG22 and FOXO3 bound to miR-629-5p. pcDNA3.1-HCG22 inhibited miR-629-5p (P<0.01), while promoted mRNA expression of FOXO3 (P<0.01). Conclusions?HCG22 was downregulated in cervical cancer, while miR-629-5p was upregulated. HCG22 gave function as a tumor suppressor in proliferation, migration and invasion of cervical cancer by targeting miR-629-5p/FOXO3 axis.     

HCG in pathologic pregnancy

Immunologic methods have transformed the field of HCG assay. Biologic and immunologic assays may give rather different results. Immunologic techniques measure the spectrum of ‘gonadotropins’ different from that detected by biologic methods. The trend of values obtained by biologic tests is usually the same as that obtained in immunoassay. In pathologic pregnancy, variations of HCG concentrations reflect changing production rates rather than altered renal excretion. A fetal metabolism of HCG occurs. In general, there are wide inter- and intrapersonal fluctations of HCG.The use of serum HCG is of prognostic value in early threatened abortion, but not in late abortion. Increased HCG excretion is not necessarily associated with hydatidiform mole pregnancy. Determination of urinary $\text{solB}\text{I}$ ratio can be a useful additional diagnostic step. Serum HCG is nearly always elevated in molar pregnancy. Serum HCG and/or urinary HCG titers are of paramount importance in the detection of choriocarcinoma following a mole, and in the clinical evaluation of patients submitted to chemotherapy. Intracranial or spinal cord metastases can be detected by HCG determinations in cerebrospinal fluid.Considerable amounts of HCG are excreted for several weeks after termination of intra-abdominal pregnancy with the placenta left in situ. There is a slowly declining production of HCG under these circumstances.Female fetuses are associated with higher HCG levels than male ones, which may suggest that the fetus exerts some control over placental HCG synthesis. In multiple pregnancy, urinary HCG excretion has been reported to be above or at the upper limit of normal values. Serum HCG is abnormally high or in the normal range.In hypertension without superimposed preeclampsia, the HCG excretion is in the normal range. The excretion values do not change prior to or at the moment of intrauterine fetal death. The balance of evidence suggests that the mean values of serum and urinary HCG are mostly higher in severe toxemia than in normal pregnancy. This is correlated with a higher placental concentration of HCG. Routine HCG assays are of no value in the clinical management of toxemia of pregnancy. In cases of severe Rh iso-immunization, the serum and urinary HCG values are increased, presumably because of the increased HCG production by the large and hydropic placenta. In terms of fetal viability, HCG determinations are of no value. In pregnancy complicated with diabetes there is no relationship between HCG levels in the body fluids and the outcome of pregnancy.In prematurity and in postmaturity, the excretion of HCG is in the normal range.     

Pregnancy-Promoting Actions of HCG in Human Myometrium and Fetal Membranes

Human chorionic gonadotropin (HCG) plays a major role in early human development through a series of well recognized pregnancy-promoting actions that are exerted in the first trimester, including maternal recognition of pregnancy, enhancement of embryo implantation and survival, stimulation of trophoblast growth and differentiation, and prolongation of the functional life of the corpus luteum. Recent research indicates that HCG can exert significant pregnancy-promoting actions also in the remainder of pregnancy through its effect on the myometrium and on fetal membranes. In the myometrium, HCG promotes the inhibition of smooth muscle cell contractility through several mechanisms, including inhibition of gap junction formation, reduction of intracellular calcium concentration, increase in the expression of progesterone receptor, and an increase in the expression of phosphodiesterase 5 (PDE5), an enzyme controlling the intracellular levels of cGMP. This effect appears to be specific for PDE5 since it has not been found for other hormones potentially involved in pregnancy such as estrogen, progesterone and thyroid hormone. In fetal membranes, HCG can modulate expression of the inducible isoform of nitric oxide synthase (iNOS), as well as specific immunoregulatory cytokines such as the high mobility group box 1 (HMGB1) protein. This accumulating evidence suggests that HCG has a wide spread pregnancy-promoting actions that are exerted in various reproductive and gestational tissues.     

子(癎)前期患者血清TPA、HCG测定的对比意义

目的:探讨子癎前期患者及正常妊娠晚期妇女血清TPA、HCG的变化特点及临床意义。方法:将51例子癎前期患者设为实验组,分为轻度(A组)、重度(B组),33例正常晚孕者设为对照组,测定实验组、对照组TPA及HCG变化,并进行比较。结果:实验组TPA的浓度与对照组比较差异有显著性(P<0.05),实验B组与对照组比较差异有非常显著性(P<0.01),且与病情严重程度呈正相关;实验组HCG浓度与对照组比较差异有显著性(P<0.05)。实验A组与对照组比较差异非常显著(P<0.01)。结论:子癎前期患者外周血TPA增高与滋养细胞凋亡增加有关,而HCG分泌增加与缺氧状态下滋养细胞反应性增生有关,提示子癎前期患者有明显的滋养细胞增生与分化异常存在。TPA作为外周血滋养细胞凋亡的标志物,可间接反映胎盘功能的变化。     

Immunocytochemical identification of mitotic Hofbauer cells in cultures of first trimester human placental villi

Summary In cultures of first trimester human placental villi, mitotic Hofbauer cells have been identified using a combined autoradiographic and immunostaining technique for the demonstration of HCG, a marker for Hofbauer cells.     

Immunohistochemical expression of subunit beta HCG in breast cancer.

The ectopic production of HCG by non-trophoblastic tumors is well documented. Adenocarcinomas arising in the mammary gland have been shown to stain positively for the beta subunit of HCG using immunoperoxidase technique. In the present study we used the Monoclonal antibodies (M. abs) H6, H34 and H54 directed against the subunits of this hormone. A total of 31 breast tissue sections from formalin fixed, paraffin embedded unselected material was examined, out of which 22 concerned different types of primary breast cancer (Ca), 5 local recurrences, 3 cystic disease (CD) and one was an axillary lymph node metastasis (LNM). All cases were evaluated for cytoplasmic localization of the beta subunit HCG using the Mab H36 and among the 27 breast Ca 9 were studied with Mab H34 and 6 with Mab H54. Our results with the Mab H6 showed the following: 1) 55.5% of the Ca including the recurrences, were strongly positive (+) and 18.5% weakly positive (+/-); 2) 63.6% of the Grade III tumors were negative (-), whereas 81.2% of the Grade II were +, 18.7% +/- and none was -; 3) It seems that by increasing infiltration of the tumor size positivity is increased; 4) Lymphocytic infiltration of the tumor stroma and LNM did not appear related; 5) From the FU data of 14 cases collected up to now, the 10-year-survival seems to be inconclusive; 6) The 3 cases of CD were negative except for the apocrine cells which were positive; 7) The unique axillary LNM was positive.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fertilization in vitro of cumulus-enclosed mouse oocytes: effect of timing of the ovulatory HCG injection

Human chorionic gonadotropin (HCG) may be injected to time ovulation and plan oocyte retrieval for clinical in vitro fertilization (IVF). Neither clinical nor experimental data on effects of subtle alterations in timing of the HCG injection on oocyte fertilizability in vitro were available. We induced follicular development in immature hybrid mice with an injection (4 IU) of pregnant mare serum gonadotropin (PMSG). In the first series of experiments, HCG was given 42, 46 or 50 hours later. We collected cumulus-enclosed, oviducal oocytes for IVF using capacitated mouse sperm 13 hours after the last HCG injection. The fertilization incidence (mean, three experiments) fell as the PMSG-HCG interval was reduced (50 hours, 64%; 46 hours, 40%; 42 hours 24%), but this could be explained by a corresponding increase in spontaneous oocyte activation caused by prolonging the HCG-oocyte collection interval. In the second series of experiments, the latter was fixed at 13 hours; the PMSG interval was altered by staggering the initial PMSG injection. No spontaneous activation occurred, but oocytes collected after "early" HCG injection still showed significantly lower fertilization incidences (42 hours, 36%; 46 hours, 46%) compared with the 50-hour injection (66%). The possible clinical implication of this finding is discussed.     

Localization of choriocarcinoma by 131I-βHCG antibody

A patient is presented who was cured surgically of metastatic gestational trophoblastic disease (GTD), of high-risk category, after the localization of tumor by radioactive antibody to HCG. The patient was initially treated with hysterectomy and resection of involved bowel; disease recurred, and conventional triple-agent chemotherapy failed, as well as less established multidrug reginens. Rabbit antibody to β-HCG was labeled with ¹³¹I and injected into the patient. Scintigrams localized the metastatic disease, allowing resection and cure. This methodology may offer a potential means of utilizing surgery to cure select patients who have failed triple-agent therapy and whose prognosis is poor if treated conventionally.     

The HCG ratio as a predictor of pregnancy outcome in assisted conception cycles

ObjectiveTo determine whether the HCG ratio can be used to predict pregnancy viability in patients undergoing IVF/ICSI treatment.Design and settingsThis was a prospective observational study conducted in a private assisted conception unit.Subjects and methodsThe patients recruited had one either a long luteal agonist protocol, a short agonist protocol, or an antagonist protocol. All patients had a maximum of three embryos transferred per cycle. Pregnancy detection was by routine serum HCG measurement on day 14 after oocyte retrieval (HCG 0) followed by another HCG sample 48h later (HCG 48). Patients with an initial positive HCG had a transvaginal ultrasound 14days later to determine viability.ResultsThree hundred and twenty patients were included in the study. We used receiver operating characteristics (ROC) analysis to predict the ability of HCG measured at 14days (HCG 0), HCG measured at 16days (HCG 48) after oocyte retrieval as well as the HCG ratio (HCG 48/HCG 0) to predict pregnancy viability as well as to predict multiple pregnancy. The HCG ratio with an optimal cut-off of 1.82 had a sensitivity of 97.6%, a specificity of 98.2% and an area under the ROC curve of 98% in the prediction of pregnancy viability. In the prediction of multiple pregnancy the HCG ratio had an optimal cut-off of 2.06 with a sensitivity of 94.5% and a specificity of only 35.6% and an area under of only the ROC curve of 64%. However, the HCG 0 with a cut-off value of 118.56mIU/ml (sensitivity 97%, specificity 96.5%) and the HCG 48 with a cut-off value of 258.16mIU/ml (sensitivity 97.2%, specificity 99.4%) were shown to be accurate in predicting a viable intrauterine multiple pregnancy with an area under the ROC curve of 97% and 99%, respectively.ConclusionThe HCG ratio with a cut-off value of 1.82 can be used to predict pregnancy viability in assisted conception cycles. Also HCG measured 14 and 16days after oocyte retrieval with a cut-off value of 118.56mIU/ml and 258mIU/ml can be used to predict viable multiple pregnancy.     

The disappearance of human chorionic gonadotropin from plasma and urine following induced abortion. Disappearance of HCG after induced abortion

In 28 females, daily measurement of the HCG concentration in urine and in 15 of them daily measurement of the beta-HCG concentration in plasma was carried out during the first 2 weeks following first-trimester induced abortion by vacuum aspiration. Plasma beta-HCG concentration fell according to a multi-exponential curve with a half-life of 0.63 days in the first 2 days following induced abortion, and of 3.85 days in the subsequent 14 days. The disappearance of HCG from urine is exponential, with a half-life value of 1.3 days. A urine pregnancy test with a sensitivity of 1 IU/ml wil nearly always be negative in the course of 2 weeks after abortion. A positive test 4 weeks after abortion indicates an incomplete abortion or persistent trophoblast.     

Mismanagement of choriocarcinoma due to a false low HCG measurement

Accurate measurement of HCG is essential for the diagnosis and management of patients with choriocarcinoma. It is therefore important that clinicians are aware of the possible analytical problems associated with the HCG assay if delay in the diagnosis of this curable tumor is to be avoided. We describe the case of a 43-year-old woman with extensive lung metastases who had an unnecessary hysterectomy and lung biopsy and in whom the diagnosis of this potentially rapidly fatal tumor was delayed because of a falsely low HCG level measured by a commercial kit. This underestimation of HCG was due to the 'high dose hook' effect, which is described in detail, and methods for avoiding this analytical problem are described.     

A comparison of pregnancy rate before and after the administration of HCG in intrauterine insemination

Purpose

Intrauterine insemination (IUI) is one of the first treatments of infertility. In natural cycles, women conceive when an intercourse takes place during a 6-day period ending on the day of ovulation. The current practice in IUI cycles is to perform IUI 24–36 h after the HCG administration, when the ovulation already took place. In this study, HCG was administered after IUI, which more closely resembles the fertilization process in natural cycles. The aim of the present study is to compare the fertility rates in an IUI protocol in women who took an HCG injection before and after the IUI.

Methods

This study was conducted on 100 infertile couples who referred to the infertility research center of Shahid Sadoughi University of Medical Sciences. They were divided into two groups: HCG injection before IUI and HCG injection after IUI. The main outcome measure was the result of a β HCG test that was done two weeks after the IUI; if it was positive, transvaginal sonography would be performed in the seventh week for clinical confirmation of pregnancy.

Results

The analysis included 50 cycles with HCG administered before and 50 cycles with HCG administered after the IUI. The pregnancy rates were 10 and 12 % (P = 0.85), respectively. Independent factor affected the cycle outcome was the time of infertility.

Conclusion

HCG administration after IUI brought about no improvement in the pregnancy rate. Therefore, HCG can be administered either before or after IUI.     

High-level HCG in non-gravidic situations: About two cases

Ovarian dysgerminoma is the most common germinal tumor in women; however, a lot of different symptoms can lead to its diagnosis. In the two cases reported here, misdiagnosis of ectopic pregnancy was first done because of inappropriate secretion of HCG by the tumor. These two cases point out the particularity of dysgerminoma with its various secretion capacity. Conversely, facing a raised level of HCG in non-gravidic situation, physicians have to consider different gynaecological and extragynaecological hypothesis.     

Low-dose HCG may improve pregnancy rates and lower OHSS in antagonist cycles: a meta-analysis

Human chorionic gonadotrophin (HCG) may substitute FSH to complete follicular growth in IVF cycles. This may be useful in the prevention of ovarian hyperstimulation syndrome. Relevant studies were identified on Medline. To evaluate outcomes, a meta-analysis of low-dose HCG-supplemented IVF cycles versus non-supplemented ones was performed with data from 435 patients undergoing IVF who were administered low-dose HCG in various agonist and antagonist protocols and from 597 conservatively treated patients who served, as control subjects. Using these published data, a decision analysis evaluated four different management strategies. Effectiveness and economic outcomes were assessed by FSH consumption, clinical pregnancy and incremental cost-effectiveness ratios. Clinical pregnancy and ovarian hyperstimulation were the main outcome measures. Nine trials published in 2002–2007 were included. From the prospective studies, in the gonadotrophin-releasing hormone antagonist group, a trend for significance in clinical pregnancy rate was evident (odds ratio [OR], 1.54; 95% confidence interval [CI], 0.98–2.42). Ovarian hyperstimulation was less significant in the antagonist low-dose HCG protocol compared with the non-supplemented agonist protocol (OR 0.30; 95% CI 0.09–0.96). Less FSH was consumed in the low-dose HCG group but this difference was not statistically significant. Low-dose HCG supplementation may improve pregnancy rates in antagonist protocols. Overall, low-dose HCG-supplemented protocols are a cost-effective strategy.     

依据扳机日血清E2值制定HCG剂量对ICSI结局的影响

目的 探讨依据扳机日血清E2值制定HCG剂量对卵胞浆内单精子注射（ICSI）促排卵结局的影响。方法 前瞻性分析2012年1－12月在上海瑞金医院生殖中心行ICSI助孕依据HCG日血清E2值制定不同HCG扳机剂量的117名患者的促排卵结局。HCG扳机剂量制定原则：E2值≤16 515 pmol/L为7000 IU、E2值介于16 516～23 854 pmol/L为5000 IU、E2值≥23 855 pmol/L为3000 IU。结果 3组MⅡ率、受精率的差异无统计学意义。7000 IU组卵裂率低于5000 IU组（96.43% vs. 99.62%）,差异有统计学意义（P<0.05）,7000 IU组可用胚胎率高于3000 IU组和5000 IU组（65.43% vs.55.69%和57.74%）,差异有统计学意义（P<0.05）。5000 IU组发生1例中度OHSS。3组新鲜移植周期的平均移植胚胎数、临床妊娠率和胚胎着床率的差异无统计学意义。结论 依据HCG日E2值制定HCG扳机剂量不影响卵母细胞成熟率和妊娠率,即高E2值予以低剂量HCG,低E2值予以高剂量HCG的原则是可行的。