人绒毛膜促性腺激素诱导JEG-3细胞TGF-β3的表达

目的:探讨人绒毛膜促性腺激素（hCG）对绒癌细胞系表达转化生长因子（TGF-β3）的影响。方法:以绒癌细胞来源的JEG-3细胞系为研究对象,采用半定量逆转录多聚酶链反应（RT-PCR）方法,观察HCG对JEG-3细胞表达TGF-β3　mRNA的影响响。结果:分别用0、50、500、5000、25000 mU/ml　hCG处理48h后,JEG-3细胞中TGF-β3　mRNA的含量逐渐被诱导增多,并表现出一定的浓度效应——随hCG作用浓度增高而显著;另外,分别检测受25 000mU/ml hCG处理0、6、12、24、30、36、48h的JEG-3细胞中TGF-β3 mRNA的表达情况,发现TGF-β3表达受hCG的诱导在30h时最强,之后逐渐减弱。结论:hCG可能通过调节TGF-β3在滋养细胞或滋养细胞来源的绒癌细胞中的表达,进而影响细胞的浸润转移。     

细胞因子对人绒毛膜组织hCG分泌的影响

综合前人经验建立一个接近于体内生理状态的体外分泌ｈＣＧ的模型，并在此基础上研究ＩＬ－１ａ，、ＴＮＦａ、ＩＦＮａ单独或两两协同对ｈＣＧ的调节作用，旨在为阐明免疫细胞因子对内分泌激素ｈＣＧ的调节作用，尤其是在生理状态下的调节作用提供一些实验依据。     

人绒毛膜促性腺激素(hCG)抑制人PBMC促炎细胞因子mRNA的表达

目的：探讨人绒毛膜促性腺激素（hCG）对促炎细胞因子基因表达是否有影响。方法：人PBMC与不同浓度的hCG（100、50、25、12．5、6．25、3．125U/ml）共同于37℃、5％CO2条件下培养4小时，培养细胞行RT－PCR测定，RT－PCR产物经琼脂糖电泳后用图象分析仪作半定量测定，比较各组结果。结果：50－6.25U／ml的hCG对TNF－αmRNA的表达有抑制作用，且与不加hCG的对照组比较有显著差异（P＜0．01）；而25及12．5U/ml的hCG对PBMC IL－1及IL－6 mRNA的表达均有明显抑制作用（P＜0．01）。结论：hCG在一定剂量范围内有抑制促炎细胞因子mRNA表达的作用，提示在正常人体中也可存在的hCG在机体促炎／抗炎自稳机制中可能参与一定作用。     

hCG-β反义寡脱氧核苷酸抑制正常滋养细胞hCG的分泌

目的：探讨ｈＣＧ－β反义寡脱氧核苷酸（ＡＳ－ＯＤＮ）是否抑制正常早期妊娠滋养细胞ｈＣＧ的分泌、作用效率及对该细胞生长的影响。方法：采用放射免疫分析法检测了与ｈＣＧ－βＡＳ－ＯＤＮ共培养后物正常滋养细胞ｈＣＧ的含量结果：ｈＣＧ－βＡＳ－ＯＤＮ能在ｈＣＧ蛋白水平产生抑制作用。结论：ｈＣＧ－βＡＳ－ＯＤＮ能牧场卉地抑制正常滋养细胞ｈＣＧ蛋白的分泌。     

IL-10对早孕滋养细胞MMP-9表达的影响

人类滋养细胞的侵入仅限于子宫内膜及肌层的浅层 1/ 3,并于妊娠中期停止[1] 。这一过程受到多种因素的控制 ,ＩＬ 10在整个妊娠过程中都可由胎盘滋养细胞分泌 ,且α6整合素表达阳性的滋养细胞表面也存有ＩＬ 10的受体 ,故ＩＬ 10也是参与该调节的因子之一[2 ] 。侵入并非一个由细胞增殖造成的被动事件 ,而是一个主动的生化过程。滋养细胞通过分泌蛋白酶类获得侵袭能力 ,其中基质金属蛋白酶 9(ＭＭＰ 9)是参与滋养细胞侵入子宫内膜的主要蛋白酶[3 ] 。因此 ,为探讨ＩＬ 10对滋养细胞侵袭力的影响 ,我们采用ＲＴ ＰＣＲ方法 ,观察ＩＬ 10对早…     

游离β-HCG检测在滋养细胞肿瘤诊断中的意义

采用放射免疫与免疫放射分析法检测了滋养细胞疾病与肿瘤患者血清中的HCG及其游离β-亚单位(F-β-HCG),计算F-β-HCG/HCG比值,并与正常妊娠标本比较。结果,妊娠组于妊娠第5周后F-β-HCG/HCG比值平均为1.18％(0.07～5.90％);葡萄胎组为4.25％(0.08～11.00％),显著高于妊娠组P〈0.005;侵蚀性葡萄胎组为7.d7％(0.30～21.20％)显著高于妊娠组与葡萄胎组,P〈0.005;绒癌组为11.25％(0.80～30.00％),显著高于妊娠组与葡萄胎组,P〈0.005。提示,在葡萄胎和滋养细胞肿瘤时F-β-HCG显著增高,检测F-β-HCG,计算F-β-HCG/HCG比值有助于滋养细胞疾病与肿瘤的诊断和鉴别诊断。     

hCGβ-C3d融合蛋白在CHO细胞中的表达

为了提高hCGβ的免疫原性 ,构建含新型分子佐剂C3d的hCGβ真核细胞表达质粒pcDNA3 hCGβ C3d3,使其转染稳定表达系统中国仓鼠卵细胞 (CHO )后 ,检测融合蛋白分泌情况。CHO细胞转染了pcDNA3 hCGβ C3d3质粒后 ,经 4 8h培养 ,1 0× 10 6个CHO细胞可分泌 6 6 0ng的重组融合hCGβ C3d3蛋白。细胞免疫化学技术分析显示 ,表达产物既有hCGβ的组分也有C3d的成分。经SDS PAGE及免疫印迹试验发现CHO细胞培养上清液中含有三组蛋白成分 ,其相对分子质量分别为12 0 0 0 0、90 0 0 0和 6 5 0 0 0。利用新型分子佐剂C3d与hCGβ的连接 ,构建了pcDNA3 hCGβ C3d3真核细胞表达质粒 ,为提高hCGβDNA免疫避孕疫苗的免疫原性及抗生育作用奠定了基础     

hCGβ和hCGβ-C3d3融合蛋白在CHO细胞中的分泌性表达、鉴定与纯化

目的：构建带有6个组氨酸纯化标签的hCGβ和hCGβ-C3d3融合蛋白的分泌型真核表达质粒，在CHO细胞中获得具有生物学活性的重组蛋白的高效表达。方法：利用高效真核表达载体phCMV1，构建phCMV1-6His-hCGβ和phCMV1-6His-hCGβ-C3d3，脂质体法转染CHO细胞，G418(800μg/ml)筛选抗性克隆。放射免疫法检测抗性克隆培液中hCGβ表达量，Western blotting和Raji细胞免疫化学法鉴定hCGβ-C3d3融合蛋白。用镍柱和凝胶过滤层析纯化表达产物。结果：酶切及测序结果显示，phCMV1-6His-hCGβ及phCMV1-6His-hCGβ-C3d3构建正确。在CHO细胞中成功地获得了hCGβ和hCGβ-C3d3融合蛋白的高效表达，phCMV1载体的表达效率是pcDNA3的1．6倍。表达产物经纯化后得到了所需的hCGβ和hCGβ-C3d3融合蛋白。结论：hCGβ和hCGβ-C3d3融合蛋白在CHO细胞的高效表达为下一步比较hCGβ抗原及hCGβ-C3d3融合蛋白免疫动物的体液免疫效应和抗生育效果奠定了基础。     

早孕滋养细胞膜型及可溶性趋化因子CXCL16表达调控

目的 探讨趋化因子CXCL16在人早孕滋养细胞表达和释放的调控机制.方法 原代培养人早孕滋养细胞,实时定量RT-PCR、免疫化学和ELISA方法分析CXCL16的表达和分泌;ELISA方法分析细胞因子IFN-γ、TNF-α、IL-4刺激前后滋养细胞CXCL16的表达和分泌水平;ELISA方法分析ADAM10治疗前后滋养细胞CXCL16的表达及分泌水平.结果 滋养细胞表达并分泌趋化因子CXCL16;IFN-γ治疗后滋养细胞表达和分泌CXCL16水平均显著上升(P<0.01),但TNF-α和IL-4并不影响CXCL16的表达或分泌;ADAM10可以加速CXCL16自滋养细胞的脱落,但并不上调CXCL16的合成.结论 IFN-γ和ADAM10参与调控母胎界面滋养细胞趋化因子CXCL16的合成和分泌.     

抗hCG单克隆抗体对hCG和LH刺激Leydig细胞分泌睾酮的影响

本工作观察了五种抗hCG单克隆抗体对hCG/LH刺激BALB/c小鼠Leydig细胞分泌睾酮的影响。抗整个hCG分子的抗体2C_5~*及抗hCH-β亚单位的抗体3C_6~*和2A_4~*对hCG及LH刺激睾酮分泌均有抑制作用。抗整个hCG分子的抗体hCG_1~*可阻断hCG刺激睾酮分泌,而对LH则无阻断作用。抗hCG-β亚单位羧基端37肽片段的抗体3A_4~*既不能抑制hCG,也不能抑制LH刺激睾酮分泌。结果提示在hCG分子上存在着特异的与其生物学活性表达有关的抗原决定簇。     

Pregnancy: Clearance curves of serum human chorionic gonadotrophin for the diagnosis of persistent trophoblast

A well recognized hazard of conservative surgical treatmentof tubal pregnancy is incomplete removal of trophoblastic tissue.Persistent trophoblast can be detected by postoperative serumhuman chorionic gonadotrophin (HCG) monitoring. The impact ofvarious surgical techniques on the post-operative clearanceof serum HCG was investigated in a retrospective study. Themedical records of 97 patients treated surgically for tubalpregnancy in the Academic Medical Center of the University ofAmsterdam, The Netherlands, between 1 January 1992 and 1 August1994 were reviewed; 28 patients were treated by salpingostomyby laparoscopy, 16 by salpingostomy by open surgery and 53 bysalpingectomy by either method. There was no difference in thepost-operative clearance of serum HCG after successful conservativesurgery compared to radical surgery. However, persistent trophoblastoccurred in eight patients (29%) after laparoscopic salpingostomyand in only one patient (6.3%) who had a salpingostomy by opensurgery (relative risk 4.57). Serum HCG clearance curves allowearly identification of patients with persistent trophoblastafter conservative surgical treatment. Moreover, monitoringof post-operative serum HCG until it becomes undetectable ismandatory in order to reveal late-onset types of persistenttrophoblast.     

人绒毛膜促性腺激素对JEG-3细胞表达VEGF的影响

目的: 揭示人绒毛膜促性腺激素(hCG)对滋养层细胞表达血管内皮生长因子(VEGF)的影响。方法: 以滋养层细胞来源的JEG-3细胞系为研究对象,采用半定量逆转录多聚酶链反应(RT-PCR)方法检测VEGFmRNA的表达。结果: 分别用(0、50、500、5000、25000)U/LhCG处理48h后,JEG-3细胞中VEGF的表达被诱导增高,且包括VEGF₁₂₁、VEGF₁₆₅、VEGF₁₈₉在内的VEGF亚型均受到诱导。另外,观察VEGF在25000U/LhCG处理的JEG-3细胞中表达的时间变化,结果发现VEGF在JEG-3细胞中的表达在经过短暂的诱导后略降低。结论: hCG可能通过调节VEGF在滋养层细胞或滋养层细胞来源的绒癌细胞中的表达,进而影响细胞的增殖、迁移或浸润转移。     

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.     

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.     

Proteomic analysis of knock-down HLA-G in invasion of human trophoblast cell line JEG-3

Previous studies showed that aberrant HLA-G expression in trophoblast cells plays important roles in trophoblast invasion; however, the mechanisms remain to be explored. In this study, we found that suppressed HLA-G expression could dramatically decrease the mRNA and protein expression levels of matrix metalloproteinase 2 and matrix metalloproteinase 9, and in the proteome assay, there were 3 identified proteins namely, prefoldin 1, eukaryotic translation elongation factor 2 and malate dehydrogenase 2, which were verified by Western blot and known to be associated with invasion, cell cycle and cell metabolism, respectively. Collectively, our study indicated a potential involvement of HLA-G in autocrine networks that may regulate prefoldin, MMPs and trophoblast invasion at the maternal-fetal interface in human pregnancy.     

Effect of chorionic gonadotrophin on hematopoietic stem cells

Department of Biological Chemistry, Perm Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR N. V. Vasil'ev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 109, No. 1, pp. 62–64, January, 1990.     

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.