男性肺原发性绒毛膜癌1例并文献复习

回顾性分析南京医科大学第一附属医院通过手术切除病理确诊的1例男性肺原发性绒毛膜癌(primary choriocarcinoma,PCC)的临床资料、影像学及组织病理学等特征,结合国内外文献复习,总结肺PCC的诊断及治疗进展.男性肺PCC是一种较为罕见高度恶性生殖细胞肿瘤,病因不明,临床表现多样,咯血最常见.早期诊断困难,易误诊为常见病,预后差,致死率高.手术联合化疗是目前较为提倡的治疗方案.男性肺PCC的诊断较难,需结合临床、影像学等综合分析,确诊需依赖组织病理学.     

Morpho-functional study of human luteal cell subpopulations

It has been reported that the mammalian corpus luteum is composedmainly of two subpopulations of luteal cells (large and small)of different morphology and function. The aims of this studywere first to characterize cytologically the human corpus luteumthroughout the luteal phase, and second to establish the in-vitrosteroidogenic capacity of a well-defined human mid-luteal cellsystem. The results show that the most predominant (>70%)cell shape, is polyhedric, and the number of cells per unitarea is significantly different in the early, mid- and latecorpus luteum (P< 0.005). Moreover, small cells (<22 µm)were most common (56.8%) in all tissues analysed. On the otherhand, both subpopulations synthesized progesterone, oestradioland testosterone, although a significantly greater productionof basal steroids was observed in large luteal cells (P<0.05). Nevertheless, the response of small cells to human chorionicgonadotrophin (HCG) was significantly greater (P< 0.05) thanthat of large cells, in agreement with the preferential specificbinding obtained for (125I)HCG to the small cell subpopulation.In summary, these results indicate that the human corpus luteumpossesses distinct cell types, which may be related to endocrinefunction and its control.     

Cell proliferation and apoptosis: Immunohistochemical evidence of p53 protein in human placenta and choriocarcinoma cell lines

We investigated the expression of the tumour-suppressor andcell cycle control protein p53 in human first trimester andterm placenta, three choriocarcinoma cell lines (Jeg-3, JAR,BeWo) and human choriocarcinoma. Using monoclonal antibodiesagainst p53 (DO-7, Ab-6, DO-1, PAb 1801), paraffin-embeddedsections of first trimester and full-term placentae, human choriocarcinomaand Jeg-3, JAR and BeWo, as well as cytospins, were evaluatedimmunohistochemically. In addition, Western blots were carriedout with the same antibodies on choriocarcinoma cell lines.In placentae, a small number of villous and extravillous cytotrophoblastcells, as well as very few syncytiotrophoblast cells, stainedintensively. Also, p53 was visualized in some nuclei of theplacental basal plate, whereas stroma and endothelium were negativefor p53. Jeg-3, JAR and BeWo also showed a positive nuclearreaction with all applied antibodies. In paraffin-embedded sectionsof human choriocarcinoma, staining was confined to the nucleiof malignant cells. The results suggest that p53 is overexpressednot only in malignant tumour cells but in certain trophoblastcell populations of the human placenta as well.     

Peripheral blood mononuclear cells in early pregnancy promote invasion of human choriocarcinoma cell line, BeWo cells

BACKGROUND: During the establishment of the maternal blood circulation around the implanting human embryo, maternal peripheral blood mononuclear cells (PBMC) directly contact trophoblasts. To determine the physiological significance of this interaction, the effects of PBMC obtained from pregnant women on the proliferative and invasive properties of a human choriocarcinoma cell line, BeWo cells, were examined. METHODS AND RESULTS: PBMC were obtained from women in early pregnancy and from women in the secretory phase of the menstrual cycle. PBMC from pregnant women significantly increased the number of invading BeWo cells in an invasion assay without affecting the proliferation of BeWo cells (P +/- 0.05). No significant changes were observed in the co-cultures with PBMC from non-pregnant women. The addition of conditioned medium, which was prepared by 2 days of incubation with PBMC from pregnant women, also enhanced BeWo cell invasion in a dose-dependent manner. Moreover, when PBMC obtained from non-pregnant women were incubated with recombinant HCG (0-10 IU/ml) for 2 days, significant augmentation of the effect on BeWo cell invasion was observed in the conditioned medium from HCG-treated PBMC (P +/- 0.05). CONCLUSION: This study indicated that soluble factor(s) secreted from PBMC promote BeWo cell invasion. It also showed the possible involvement of HCG in the regulation of BeWo cell invasion by PBMC. These findings suggest crosstalk between maternal PBMC and trophoblasts via soluble factor(s), which may play an important role in early embryo implantation.     

Comparative stimulatory effect of gonadotrophin releasing hormone (GnRH) and GnRH agonist upon pulsatile human chorionic gonadotrophin secretion in superfused placental explants: reversible inhibition by a GnRH antagonist.

The roles of gonadotrophin releasing hormone (GnRH) and a GnRH agonist (GnRHa) (D-Ala6-Met-Leu7-Pro-N-ethyl-amide) in controlling pulsatile human chorionic gonadotrophin (HCG) secretion by superfused placental explants in the first trimester were examined. One minute pulses of both GnRH and GnRHa had a biphasic effect upon pulsatile HCG secretion. GnRHa was maximally effective at 10(-10) M concentration, at 10(-11) M the effect was mild while at 10(-8) M, no effect was noted. GnRH exerted a maximal stimulatory effect at 10(-8) M; at 10(-10) M no effect was seen, while at 10(-7) M the effect was mildly stimulatory. This was evaluated by carrying out both a between and within channel type of analysis. The effect of a GnRH antagonist GnRH(ant) upon GnRH and GnRHa-induced HCG secretion was examined. Explants were incubated overnight with 10(-8) M GnRH(ant), which was also continuously administered during superfusion. The addition of 1-min pulses of GnRH and GnRHa during the exposure to GnRH(ant) failed to stimulate pulsatile HCG secretion. This effect was reversible since the response to GnRH was restored within 10 min after stopping GnRH(ant) administration. In addition, by the third cycle, co-administration of GnRH(ant) for 2 min together with 10(-10) M GnRHa for 1 min completely blocked the GnRHa-induced effect. Continuous administration of 10(-8) M GnRH(ant) decreased spontaneous HCG pulse amplitude and the area under the curve but failed to modify pulse frequency. In conclusion, GnRH appears to exert a receptor-dependent stimulatory effect upon pulsatile HCG secretion in superfusion in the first trimester placenta.(ABSTRACT TRUNCATED AT 250 WORDS)     

Endometrial stromal cell decidualization inhibits human chorionic gonadotrophin and human placental lactogen secretion by co-cultured trophoblasts

We have developed an in-vitro co-culture system to examine theinteraction between purified first trimester cytotrophoblastsand purified non-pregnant human endometrial stromal cells (ESC).ESC decidualization is an important step in endometrial maturationand may modulate embryo implantation. In order to investigatethe effects of ESC decidualization on trophoblast function,we examined human chorionic gonadotrophin (HCG), human placentallactogen (HPL), progesterone and oestrogen secretion by trophoblastsco-cultured in contact with ESC, either with or without decidualizationinduced by progesterone. Decidualized ESC inhibited basal HCGand HPL secretion for 3 days during the culture for HCG, andfor 5 days during the culture for HPL (P < 0.01 and P <0.03 respectively). After 5 days of co-culture, decidual transformationof ESC as indicated by prolactin production occurred in thecontrol cultures due to progesterone and oestradiol secretionby the co-cultured trophoblasts, but no significant differencesin HCG or HPL secretion were observed between the two groups.Although the type of trophoblast used in the present study isfar from implantation, our results clearly demonstrated thatHCG and HPL secretion by trophoblasts was inhibited by the presenceof co-cultured decidualized ESC, and suggested that ESC decidualizationmay regulate trophoblast function at the human fetal-maternalinterface.     

Pregnany: Human chorionic gonadotrophin release and tissue viability in placental organ culture

The use of human chorionic gonadotrophin (HCG) secretion asa measure of viability during the organ culture of human firsttrimester placental tissue has become a popular practice. Ithas been suggested that if cultured tissue is releasing largeamounts of this protein hormone, there is a high level of viability.We have found, however, that the cytosolic enzyme L-lactatedehydrogenase is released into the culture supernatant in asimilar daily pattern as HCG, suggesting that tissue disruptionmay be occurring, resulting in some of the observed hormonerelease. In addition, we have shown that the uptake of the fluorescentdye dansyl-L-lysine into the syncytium increases significantlyfrom day 0 to day 4, suggesting a loss of syncytial membraneintegrity. Electron micrographs show further evidence of thesyncytial degeneration at the ultrastructural level, displayingextensive vacuolation and poor micro-villous cover. In contrastto the degenerated state of the syncytiotrophoblast, a highlevel of bromodeoxyuridine incorporation is observed for cytotrophoblastsand, in particular, stromal cells up to 5 days in culture. Overall,the results suggest that the use of HCG release as a determinantof tissue viability in placental organ culture should be treatedwith a degree of caution.     

Inwardly rectifying currents in human term placental cells

Using the whole cell voltage clamp technique, inwardly rectifying currents were observed in most cells isolated from human term placentas. Reversal potentials, estimated by extrapolation, as well as those estimated by tail currents, were found to be between –80mV and –90mV. These potentials are close to the potassium equilibrium potential (theoretical value: –82 mV). In addition, the inwardly rectifying currents were blocked by 1 mM CsCl. The elevation of [K]e increased the current amplitudes. Furthermore, the holding currents were inwardly shifted when the holding potential was kept at –70mV. The extrapolated reversal potentials changed linearly on a semi-logarithmic graph with a slope of 57 mV/decade of [K]e. This value is close to the theoretical calculation (58 mV/decade). Consequently, it is suggested from these studies, that inwardly rectifying currents observed in human term placental cells are carried mainly by potassium ions.     

In-vivo and in-vitro assessment of the influence of ritodrine and oxytocin on the placental secretion of human chorionic gonadotrophin and placental lactogen

The purpose of this study was to evaluate, in vivo and in vitro,the influence of ritodrine and oxytocin on the placental releaseof human chorionic gonadotrophin (HCG) and placental lactogen(HPL). The in-vivo study was performed on maternal sera collectedbefore and 1 h after the onset of either ritodrine treatment(50µg i.v/min; administered to 15 women at risk of prematurelabour) or oxytocin infusion (2 mU i.v./min; administered to21 women for acceleration of slow labour). The in-vitro studywas performed on human term placental explants incubated inthe presence of 4–400 ng ritodrine/ml or 15–1500UiV oxytocin/ml. HCG and HPL were measured by radio-immunoassayon maternal sera and incubation media. Maternal circulatingconcentrations of HCG and HPL remained unaffected after 1 hof ritodrine or oxytocin treatment The in-vitro release of HCGand HPL by placental explants was not modified when ritodrineor oxytocin was added to the incubation media. The lack of influenceof ritodrine and oxytocin on the placental secretion of HCGand HPL suggests that β₂-adrenergic and oxytocin receptorsare not involved in the releasing process.     

Physiology: The production of placental protein 14 by human uterine tubal epithelial cells in culture

Cells prepared from the mucosal layer obtained from the fimbrial,proximal ampullary and distal ampullary regions of the humanuterine (Fallopian) tube have been grown in monolayer culture.Immunocytochemistry with anti-cytokeratin, anti-vimentin oranti-CD 45 antibodies indicated that the overwhelming numberof cells were epithelial in nature and were free of fibroblastsand leukocytes. Basal and steroid-stimulated placental protein14 (PP14) production was investigated in tissue obtained fromnine patients undergoing hysterectomy, by addition of oestradioland/or progesterone to confluent cultures. Basal PP14 productionvaried considerably between experiments, probably due to differencesbetween individuals from whom the tissue had been obtained.However, there was no difference in basal PP14 production bycells prepared from the fimbrial, proximal ampullary and distalampullary parts of the tube obtained from the same patient.When total PP14 production by cells obtained from an individualuterine tube was pooled both progesterone and oestradiol significantly(P < 0.05) stimulated the production of PP14 but the effectof progesterone either alone or in the presence of oestradiolwas numerically greater than that of oestradiol alone. ConsideringPP14 production by cells prepared from the different regionsof the tube showed that cells from the fimbrial region weremore responsive to steroid stimulation than cells prepared fromeither the proximal or the distal ampullary regions. All combinationsof hormonal supplementation stimulated PP14 production by cellsfrom the fimbrial region on all days measured (P < 0.05 -P < 0.001). In contrast oestradiol alone had no effect onPP14 production by cells prepared from the proximal and distalampullary parts of the tube on any day measured. Maximum oestradiol-stimulatedPP14 values, expressed as percentages of control, were (mean± SEM) 146 ± 11, 113 ± 7.5 and 108 ±8.5 for cells prepared from the fimbrial, proximal ampullaryand distal ampullary portions of the tube respectively. Maximumprogesterone or progesterone plus oestradiol-stimulated valueswere very similar; those for progesterone plus oestradiol were(mean ± SEM) 151 ± 8.6, 130 ± 6.2 and 118± 10 for cells from the fimbrial, proximal ampullaryand distal ampullary parts of the tube. The results show thata cell culture system has been established which allows theinvestigation of the function of human uterine tubal cells inculture. Using this system we have shown that PP14 productionby these cells is stimulated by steroids and that cells preparedfrom the fimbrial region of the uterine tube appear more responsiveto steroid stimulation than those prepared from other regionsof the tube.     

hCG对JEG-3滋养细胞系表达细胞因子的影响

目的：揭示人类绒毛膜促性腺激素（hCG)是否通过改变细胞因子的生成而影响滋养细胞的侵袭性。方法：以永生化的滋养细胞系JEG－3为研究对象，采用逆转录多聚酶链反应（RT－PCR）方法观察了hCG对JEG－3细胞与细胞侵袭力调节有关的多种因素因子表达的影响，结果：JEG－3细胞表达HGF，IGF－II，VEGF和TGF－β3，且VEGF的表达以VEGF121和VEGF165为主，而不表达IGF，TGF－1β，TGF－β2，IL－β1，25U／mL hCG处理50h可显著降低JEG－3细胞中HGF的表达，同时强烈诱导VEGF121和VEGF165的表达，而其它基因的表达未发生明显变化，结论：HGF对滋养细胞的侵入起促进作用，而VEGF则具有抑制效应，说明，高浓度的hCG可能通过两种细胞因子的自分泌机制对滋养细胞的侵入起抑制作用。     

Predictive value of early human chorionic gonadotrophin serum profiles for fetal growth retardation.

The possibility of using first trimester maternal serum human chorionic gonadotrophin (HCG) profiles to predict fetal growth retardation (FGR) was tested in 236 women with singleton pregnancies obtained after in-vitro fertilization (IVF). Pregnancies were monitored by serial analysis (two or more) of serum HCG at at least 48 h intervals. Serum was obtained between the 13th and the 35th day after conception (i.e. on the day of IVF). Early miscarriage occurred in 23.7% and FGR in 10.9% of pregnancies. Serum HCG profiles were higher than the 90th and lower than the 10th percentile in 12.3% and 19.5% of the cases respectively. FGR was significantly more frequent in women with serum HCG profiles lower than the 10th percentile than in women with normal profiles (45.5% versus 7.2%; P < 0.001), with a relative risk of 6.5 (95% confidence interval 2.7-15.6). FGR rates were similar in women with normal and high profiles of serum HCG. Pre-eclampsia and premature delivery rates were similar in women with normal and abnormal profiles of serum HCG. First trimester serum HCG should be further investigated as a potential marker of FGR.     

人绒毛膜促性腺激素单克隆抗体的研制

目的获得针对人绒毛膜促性腺激素单克隆抗体。方法hCG蛋白免疫Balb／c小鼠,取其脾细胞与同系小鼠骨髓瘤细胞NS-1按8：1比例融合,间接ELISA法筛选阳性克隆,有限稀释法进行克隆化培养;制备腹水抗体;采用间接ELISA法鉴定抗体亚型和测定抗体效价。结果得到6株能稳定分泌单克隆抗体的杂交瘤细胞株;抗体经鉴定均为IgG1、κ型,效价均达10^-5以上。结论所获得的6株杂交瘤细胞株均有较强稳定分泌抗-hCG单克隆抗体的能力。这为有关hCG的检测、hCG本身相关研究以及避孕疫苗的研制打下了基础。     

Molecular interactions during pregnancy: Expression of leukaemia inhibitory factor (LIF) receptor in human placenta: a possible role for LIF in the growth and differentiation of trophoblasts

Leukaemia inhibitory factor (LIF) is a cytokine that displaysmultiple activities in various tissues and is essential forblastocyst implantation in mice. In the human uterus, LIF isexpressed in endometrial tissue and the decidua. To elucidatethe role it plays, the mRNA levels for two LIF receptor (R)subunits, LIF-R and gp130, were examined in human endometrium,placenta and decidua by Northern blot hybridization. The expressionof LIF-R gene was detected in the chorionic villus during thefirst trimester, in term placenta, and at lower levels in thedecidua. The expression of LIF-R gene was not detectable innon-pregnant endometrium. The expression of the gp130 gene wasdetected in all tissues examined. During pregnancy, there wasno significant change in the mRNA concentration of LIF-R inthe placenta, while that of gp130 increased after the secondtrimester. The human choriocarcinoma cell line, BeWo, was foundto express LIF-R and gp130. LIF inhibited forskolin-inducedhuman chorionic gonadotrophin (HCG)-B production by BeWo ina dose-dependent manner, and it ameliorated forskolin-inducedgrowth suppression. These findings suggest that LIF plays aregulatory role in trophoblast growth and differentiation duringpregnancy in human placenta.     

Induction of macrophage migration inhibitory factor in human ovary by human chorionic gonadotrophin

The role of macrophage migration inhibitory factor (MIF) in human ovarian function remains obscure. The aim of this study was to investigate how MIF was related to ovulation by quantitative analysis of serum, follicular fluid and culture medium of granulosa cells obtained from in-vitro fertilization (IVF) and embryo transfer patients. Serum MIF concentrations in ovarian stimulation cycles for IVF-embryo transfer were higher at day 1 (median 92.6 ng/ml), which took place 35 h after human chorionic gonadotrophin (HCG) administration and just before the retrieval of oocytes, than those before day -6 (12.1 ng/ml), at day -5 to about day 0 (17.5 ng/ml) or at day 2 to about day 14 (8.2 ng/ml). MIF concentrations in the follicular fluid (113.4 ng/ml) obtained in ovarian stimulation cycles for IVF-embryo transfer were significantly higher than in serum (72.0 ng/ml) collected at the same time. MIF concentrations in the follicular fluid in natural cycles were higher in the ovulatory phase (51.6 ng/ml) than in the late follicular phase (13.8 ng/ml). MIF concentrations in the culture media of granulosa cells increased from 3.2 ng/ml to 7.2 ng/ml with HCG stimulation, and decreased from 2.4 ng/ml to 1.2 ng/ml when stimulation was withheld. These results indicate that HCG can induce the elevation of serum and follicular fluid MIF concentrations through the stimulation of ovarian cells, and that MIF is probably involved in the mechanism of ovulation.     

Diffuse pagetoid squamous cell carcinoma of the esophagus combined with choriocarcinoma and mucoepidermoid carcinoma: an autopsy case report

Esophageal squamous cell carcinoma in situ (SCCIS) with diffuse pagetoid features is a recently recognized rare variant of squamous cell carcinoma. A histopathological study of a specimen from a 70-year-old male Japanese patient is reported. The patient died of respiratory failure due to rapidly progressing metastatic pulmonary tumors of unknown origin 73 days after the onset of hemosputum. Autopsy disclosed widespread metastasis of choriocarcinoma in the absence of tumors of the testes or other common sites of germ cell tumors. Elevation of human chorionic gonadotropin (hCG-beta) levels was later detected in the stored serum. Serial histological evaluation of the entire esophagus revealed a small primary site of choriocarcinoma in a background of diffuse SCCIS, mainly of pagetoid type, accompanied by several small foci of submucosally invasive squamous cell carcinoma and primary mucoepidermoid carcinoma. These stimulated nodal metastasis independently of the choriocarcinoma. The SCCIS did not alter the gross mucosal appearance. This is the first reported case of diffuse pagetoid SCCIS combined with choriocarcinoma. Morphological findings and previous studies suggest that the extensive SCCIS of the esophagus resulted from pagetoid spread of tumor cells. The invasive squamous cell carcinoma, mucoepidermoid carcinoma and choriocarcinoma are suggested to have originated from the overlying SCCIS.     

Ultrastructure of epithelial plaque formation and stromal cell transformation by post-ovulatory chorionic gonadotrophin treatment in the baboon (Papio anubis).

BACKGROUND: To understand factors controlling endometrial responses to pregnancy, we have established a model using the baboon and examined the effects of infused human chorionic gonadotrophin (HCG) on the preparation of the luminal epithelium and stromal cell differentiation for the establishment of pregnancy. METHODS: The ultrastructure of endometrium from normal day 10 post-ovulation animals, cycling females treated with either HCG or FSH (control), and a day 15 pregnant animal has been compared. RESULTS: In the control endometrium, the luminal epithelium was smooth and regular, with underlying spindle shaped stromal cells. In pregnancy, the luminal epithelium underwent a plaque reaction, while stromal cells enlarged and developed filament-rich cell processes. Infusion of HCG produced changes similar to those seen in pregnancy, with generalized plaque formation and stromal decidualization, while in the animal treated with FSH there was no response. CONCLUSIONS: This study indicates that infusion of HCG into the uterus can duplicate many of the responses of the endometrium to pregnancy, although in this case the plaque reaction involved the whole of the luminal epithelium, rather than only the implantation site as in pregnancy.     

Pregnancy: Temporal relationship between the human chorionic gonadotrophin peak and the establishment of intervillous blood flow in early pregnancy

The purpose of this study was to investigate the temporal relationshipbetween the early pregnancy peak of circulating human chorionicgonadotrophin (HCG) concentration and the establishment of maternalblood flow in the placental intervillous space. The presenceof blood flow echoes within intervillous space was determinedby colour Doppler imaging from 44 women with clinically uncomplicatedpregnancy between 6 and 18 weeks gestation. Circulating HCG,free - and HCG subunits, oestradiol and progesterone concentrationswere immunoassayed in blood samples collected at the time ofDoppler examination. A continuous intervillous blood flow wasdetected in all cases with a gestational age 11.7 weeks (n =18) but never before this time. Circulating concentrations offree HCG, oestradiol and progesterone were linearly or exponentiallycorrelated with gestational age (r = 0.860, 0.903 and 0.538respectively, all with P < 0.001), indicating steady increaseof these hormones with advancing gestation. However, the bestfitted lines were found to be parabolic for HCG (r = 0.771,P < 0.001) and HCG (r = 0.695, P < 0.001), their highestpoints corresponding to 11.24 and 10.74 weeks gestational agerespectively. The close temporal relationship between the Doppleradvent of intervillous maternal blood flow and the HCG peaksuggests that the establishment of the intervillous blood flowis associated with the decline in circulating HCG concentrations.     

Ovarian steroidogenic response to human chorionic gonadotrophin in obese women with polycystic ovary syndrome: effect of metformin.

BACKGROUND: The aim of the present study was to investigate the steroidogenic response pattern to HCG in obese women with polycystic ovary syndrome (PCOS) and the possible effects of metformin treatment on it. METHODS: A single injection of human chorionic gonadotrophin (HCG, 5000 IU) was given to 12 obese [body mass index (BMI) > or = 27 kg/m2] women with PCOS and to 27 control women. Blood samples for assays of 17alpha-hydroxyprogesterone (17-OHP), androstenedione, testosterone and oestradiol were collected at baseline and 1, 2 and 4 days after the injection. Responses to HCG were also assessed in the PCOS women after 2-month treatment with metformin (500 mg x 3 daily). RESULTS: Serum 17-OHP and oestradiol concentrations peaked at 24 h in the PCOS women and preceded the maximum testosterone concentration, which was seen at 48 h. In the control women the maximum concentrations of all these steroids were reached 96 h after HCG. After metformin treatment, the basal serum testosterone concentration and the areas under the androstenedione (AUC(A)) and testosterone (AUC(T)) response curves after HCG decreased significantly. CONCLUSIONS: The results demonstrate that obese PCOS women have a male-type steroidogenic response pattern to a single injection of HCG and a higher androgen secretory capacity than control women, which may be explained by the increased thecal cell activity in the polycystic ovary. The slight alleviation of hyperandrogenism brought about by metformin therapy appears to be due to its effect on ovarian steroidogenesis possibly mediated by decreased insulin action.