血管内皮祖细胞的体外扩增特性研究

目的:探讨血管内皮祖细胞(EPCs)在体外扩增特性。方法:利用磁性活化细胞分选系统(MACS)系统富集CD34⁺细胞,在相同条件下与同批的单个核细胞(MNC)、CD34⁺和CD34^-混和细胞进行对照培养,比较EPCs体外扩增效果。另外研究血管内皮生长因子(VEGF)和传代培养对细胞分化、扩增动力学和细胞凋亡的影响。应用细胞免疫化学和流式细胞术对细胞定性定量分析。结果:MNC培养、CD34⁺和CD34^-细胞混和培养明显高于CD34⁺细胞单独培养EPCs扩增率(P＜0.05),一旦细胞形成线索样结构行传代培养明显低于未传代的细胞凋亡(P＜0.05)。VEGF对细胞凋亡(P＞0.05)无明显影响,这些分化的EPCs免疫细胞化学染色CD34、vWF、KDR、CD31阳性,并且吞噬乙酰化低密度脂蛋白(Ac-LDL)。培养7d流式细胞检查CD34⁺、AC133^＋分别占贴壁(AT)细胞的68.2%±6.3%(n=6)、57.2%±9.8%(n=6)。结论:MNC培养、CD34⁺和CD34^-细胞混和培养提高了EPCs扩增率,早传代使凋亡率明显降低。VEGF对EPCs体外扩增无明显影响。     

EPCAM、CD44和CD24在胃癌组织中的表达及其临床意义

目的: 联合检测95例胃癌组织中上皮细胞黏附分子(EPCAM)、白细胞分化抗原 44(CD44)和白细胞分化抗原 24(CD24)的表达情况,分析这3种蛋白与胃癌临床病理资料及预后之间的关系。方法: 应用免疫组织化学法检测95例经手术切除并有明确病理诊断为胃癌的标本中EPCAM、CD44和CD24的表达。分析95例胃癌临床病理资料与这3种蛋白阳性表达之间的关系。结果: (1) EPCAM阳性56例(58.95%),CD44阳性41例(43.16%),CD24阳性56例(58.95%)。其中EPCAM⁺CD44⁺30例(31.58%),EPCAM⁺CD24⁺45例(47.37%),CD44⁺CD24⁺32例(33.68%),EPCAM⁺CD44⁺CD24⁺25例(26.32%)。(2)EPCAM与年龄、肿瘤浸润深度、WHO组织学分型有关;CD44与BORRMANN分型、WHO组织学分型、CEA值有关;CD24与浸润深度、肿瘤位置、WHO组织学分型、脏器侵犯有关;三者阳性与浸润深度、肿瘤位置、WHO组织学分型有关(P<0.05)。(3)EPCAM、CD44阳性组的生存率与阴性组比较差异有统计学意义(P<0.05及P<0.01);EPCAM⁺CD44⁺CD24⁺与EPCAM^-CD44^-CD24^-的生存率比较差异有统计学意义(P<0.05); EPCAM^-CD44⁺CD24⁺与EPCAM^-CD44^-CD24^-的生存率比较有统计学意义(P<0.05)。结论: EPCAM、CD44和CD24在胃癌组织中表达阳性率高,可作为胃癌诊断的初筛实验。     

BCL-6在经典型霍奇金淋巴瘤细胞株L428和过表达CD99的L428中的表达差异及其意义

目的: 探究BCL-6在经典型霍奇金淋巴瘤(classical Hodgkin's lymphoma,cHL)细胞株L428和过表达CD99的L428(L428-CD99⁺)中的表达差异及其意义。方法: 应用免疫细胞化学和免疫荧光共聚焦检测BCL-6和CD99在cHL细胞株L428和L428-CD99⁺中的表达;运用实时荧光定量PCR和Western blotting检测细胞株L428和L428-CD99⁺中BCL-6和CD99的表达水平;MTT实验检测L428和L428-CD99⁺细胞增殖能力的差异;流式细胞术检测L428和L428-CD99⁺细胞的凋亡差异。结果: 免疫细胞化学和免疫荧光共聚焦显示CD99在L428细胞中为阴性表达,在L428-CD99⁺细胞中为阳性表达,并定位于细胞膜;BCL-6在L428细胞为阴性表达,在L428-CD99⁺细胞株中为阳性表达,并定位于细胞核。Western blotting显示CD99和BCL-6在L428细胞为阴性表达,在L428-CD99⁺细胞为阳性表达。实时荧光定量PCR结果显示,与L428细胞相比,CD99和BCL-6 mRNA在L428-CD99⁺细胞中的表达量增高(P<0.01)。MTT显示L428细胞增殖能力强于L428-CD99⁺细胞(P<0.01);流式细胞术显示L428-CD99⁺细胞凋亡较L428细胞增多(P<0.01)。结论: 过表达CD99诱发BCL-6表达增高,可导致L428细胞增殖能力下降及凋亡增加。     

地西他滨联合丙戊酸钠促进胃癌MGC-803细胞凋亡和G0/G1期阻滞的机制研究

目的: 探讨地西他滨(DCA)和丙戊酸钠(VPA)联用对胃癌细胞株MGC-803凋亡和细胞周期的影响及机制。方法: DCA 1.5 μmol·L^-1、DCA 3.0 μmol·L^-1、VPA 1.5 mmol·L^-1、DCA 1.5 μmol·L^-1+VPA 1.5 mmol·L^-1和DCA 3.0 μmol·L^-1+VPA 1.5 mmol·L^-1作用MGC-803细胞72 h。Annexin V/PI法检测细胞凋亡,流式细胞术检测细胞周期,实时荧光定量PCR检测nm23-H1 mRNA表达。结果: VPA 1.5 mmol·L^-1+DCA 1.5 μmol·L^-1联合用药组 和VPA 1.5 mmol·L^-1+DCA 3.0 μmol·L^-1联合用药组 凋亡率均高于其相应单药组,差异有统计学意义(P<0.01)。MGC-803细胞G₀/G₁期比例在VPA 1.5 mmol·L^-1+DCA 1.5 μmol·L^-1联合用药组(61.55%±2.38%)和VPA 1.5 mmol·L^-1+DCA 3.0 μmol·L^-1联合用药组(66.75%±2.48%)的表达水平均高于其相应单药组,差异有统计学意义(P<0.01)。nm23-H1 mRNA在VPA 1.5 mmol·L^-1+DCA 1.5 μmol·L^-1联合用药组(1.84±0.46)和VPA 1.5 mmol·L^-1+DCA 3.0 μmol·L^-1联合用药组(2.88±0.42)的表达水平均高于其相应单药组,差异有统计学意义(P<0.01)。结论: DCA联合VPA能显著促进MGC-803细胞凋亡及G₀/G₁期阻滞,其机制可能与促nm23-H1基因的表达有关。     

肺癌干细胞中存在容积激活氯通道

目的: 研究肺癌干细胞中是否存在容积激活氯通道。方法: 用免疫磁珠法分选非小细胞肺癌细胞系A549中CD133⁺细胞,用流式细胞术检测其纯度。用全细胞膜片钳记录CD133⁺细胞中是否存在容积激活氯电流。结果: 本实验中免疫磁珠法分选出的CD133⁺肺癌干细胞,流式细胞术检测其纯度为92.14%;在22/40(55%) CD133⁺细胞上记录到容积激活氯电流。结论: CD133⁺肺癌干细胞中存在容积激活氯通道。     

慢性乙肝患者外周血CD4CD25FOXP3 Tregs及HBV抗原特异性CTLs的检测和分析

目的: 检测慢性乙肝(CHB)患者外周血中CD4⁺CD25⁺FOXP3⁺调节性T淋巴细胞(Treg细胞)和乙肝病毒(HBV)特异性细胞毒性T淋巴细胞(CTLs)的表达及意义。方法: 收集28例CHB患者和15例健康人外周血单个核细胞标本,运用流式细胞仪对Treg细胞亚群进行定量分析,同时采用酶联免疫斑点法检测HBV抗原特异性CTLs,并结合丙氨酸氨基转移酶(ALT)和 HBV DNA的临床情况进行分析。结果: CHB组CD4⁺CD25⁺FOXP3⁺ Treg细胞的频率显著高于健康对照组 (3.14%±0.97% vs 1.95%±0.68%,P＜0.05);HBV抗原特异性CTL斑点计数为阳性(19.28±3.85)。CHB组Treg的频率与乙肝病毒载量呈正相关(r=0.831, P＜0.01),与HBV特异性CTL斑点计数值呈负相关(r＝-0.540,P＜0.01)。结论: CHB患者外周血CD4⁺CD25⁺FOXP3^＋Treg细胞表达升高并与病毒载量相关,而与HBV反应的CTLs数量呈负相关,提示Treg细胞可通过抑制细胞免疫反应影响病毒清除。     

高血压前期循环内皮祖细胞数量和功能的性别差异

背景：目前对高血压前期循环内皮祖细胞数量和功能的性别差异尚不明确。 目的：探讨性别差异对高血压前期循环内皮祖细胞数量和功能的影响。 方法：选择79例年龄(46.4±4.5)岁的人群作为研究对象,其中绝经前健康女性志愿者18例、绝经前高血压前期女性患者21例、健康男性志愿者21例和高血压前期男性患者19例,取外周血用流式细胞仪测定CD34和 KDR双标阳性循环内皮祖细胞水平,ac-LDL 及 lectin 荧光标记方法评估体外培养内皮祖细胞数量, MTT法和Transwell小室评估内皮祖细胞的增殖能力和迁移能力,测定各组血浆雌激素水平。 结果与结论：与高血压前期男性患者和健康男性志愿者比较,绝经前高血压前期女性患者和绝经前健康女性志愿者的CD34+/KDR+双阳性循环内皮祖细胞数量、ac-LDL及lectin 抗体双阳性内皮祖细胞数量增加      (P均< 0.05),内皮祖细胞迁移和增殖能力明显增强(P < 0.05),血浆雌激素水平增高(P < 0.05)。绝经前高血压前期女性患者与绝经前健康女性的内皮祖细胞迁移和增殖能力无明显差异(P > 0.05),而高血压前期男性患者的内皮祖细胞迁移和增殖能力较健康男性减弱。血浆雌激素水平与循环内皮祖细胞数量及功能呈明显的直线相关关系(P < 0.05)。这些结果证实,高血压前期循环内皮祖细胞数量和功能存在性别差异,相比高血压前期男性患者,绝经前高血压前期女性患者循环内皮祖细胞数量增多,增殖和迁移能力增强,其可能与机体内较高的雌激素水平有关。     

siRNA沉默转酮醇酶样蛋白1基因对人肝癌HepG2细胞生长微环境的影响

目的:通过siRNA技术沉默肝癌细胞系HepG2转酮醇酶样蛋白1(TKTL1)基因后观察其对生长微环境中乳酸生成水平、还原型谷胱甘肽(GSH)水平及NADPH/NADP⁺比值的影响,旨在探讨肿瘤细胞TKTL1表达与其转移特性的内在联系。方法:用构建的靶向TKTL1基因siRNA干扰质粒载体转染HepG2细胞株,RT-PCR检测TKTL1mRNA表达并结合转酮醇酶(TKT)活性测定判断其干扰效率;以未转染HepG2细胞为对照,进一步观察TKTL1沉默的癌细胞内乳酸生成水平、GSH含量、NADPH/NADP⁺比值及葡萄糖-6-磷酸脱氢酶(G-6-PD)活性的变化。结果:siRNA沉默HepG2细胞TKTL1基因后TKTL1mRNA表达下降、TKT活性显著降低(P<0.01),乳酸生成、GSH水平及NADPH/NADP⁺比值较对照组细胞均显著降低(P<0.01),但G-6-PD活性无明显变化(P＞0.05)。结论:肿瘤可能通过TKTL1高表达调整葡萄糖代谢从而改变乳酸及氧化自由基生成,使细胞生长微环境利于肿瘤转移。     

Th17细胞联合CD3+CD8-IL-21+T细胞升高与宫颈癌发生发展的关系

目的: Th17细胞在免疫调节中起重要作用,而IL-21与Th17在分化调节和功能行使上密切相关。本研究旨在探讨Th17在宫颈癌发生发展中的作用。方法: 选取37例宫颈癌患者、25例宫颈上皮内瘤变(CIN)患者和18例健康志愿者作为研究对象,用流式细胞分析术检测外周血中Th17细胞及CD3⁺CD8^-IL-21⁺T细胞的比例。分析两者与临床病理指标之间的关系。结果: 与健康对照组相比,Th17细胞及CD3⁺CD8^-IL-21⁺ T细胞比例(占淋巴细胞百分比)在CIN组(P<0.01,P<0.05)及宫颈癌组(P<0.01,P<0.05)均明显升高。此外,2种细胞的比例都与临床分期有关,在晚期宫颈癌组明显升高(均P<0.05),并且有淋巴结转移组或脉管浸润组都明显高于相对应的无转移组(P<0.01, P<0.05)或无浸润组(均P<0.01)。此外,在健康对照组和宫颈癌组,Th17与CD3⁺CD8^-IL-21⁺ T细胞呈正相关,CD3⁺CD8^-IL-21⁺ T细胞的比例还与肿瘤大小有关(P<0.01)。结论: Th17和CD3⁺CD8^-IL-21⁺ T细胞在宫颈癌患者外周血中的比例上调,在宫颈癌的发生发展中可能起着重要作用。     

母胎界面树突状细胞CCL17和CCL22表达在母胎免疫耐受中的作用

目的: 检测非孕期子宫内膜和正常妊娠早期及复发性自然流产患者蜕膜中树突状细胞(DC)CCL17和CCL22的表达差异,探讨母胎界面DC在CD4⁺CD25⁺调节性T细胞(Treg)的募集及母胎免疫耐受微环境形成中的作用。方法: 正常早孕组人工流产时、复发性流产组清宫时取其蜕膜,正常未孕组行子宫切除时取其内膜组织。分离蜕膜或子宫内膜单个核细胞,体外诱导培养DC,用real-time PCR法分析3组DC CCL17和CCL22 mRNA的表达水平,ELISA法检测3组DC培养上清液中CCL17和CCL22蛋白的表达。结果: 正常早孕组蜕膜DC CCL17和CCL22 mRNA的表达分别为3.04±0.40和1.83±0.24,均高于正常未孕组(0.85±0.24和0.31±0.08,P<0.01)和复发性流产组(1.65±0.14和0.96±0.09,P<0.01)。正常早孕组蜕膜DC能够持续旺盛分泌趋化因子CCL17和CCL22,在培养的12~96 h内CCL17和CCL22的分泌量逐渐增多。同一时点早孕组DC分泌的CCL17和CCL22均明显高于未孕组和复发性流产组DC分泌的CCL17和CCL22(P<0.01)。结论: 正常妊娠后蜕膜DC表达CCL17和CCL22增强,DC可能通过高表达CCL17和CCL22而增强对CD4⁺CD25⁺Treg的趋化作用,从而在母胎界面的免疫耐受中发挥重要作用,蜕膜DC表达CCL17和CCL22下降可能与复发性自然流产的发病有关。     

槲皮素改善大鼠骨髓来源内皮祖细胞生物学功能及其机制研究

目的:探讨槲皮素(quercetin,QUE)对大鼠骨髓来源内皮祖细胞(endothelial progenitor cells,EPCs)生物学功能的影响与机制。方法:密度梯度离心法分离大鼠骨髓单个核细胞,条件培养基EGM-2诱导分化后,进行双荧光染色及免疫表型鉴定。将培养14 d的细胞用PI3K抑制剂BYL719(3μmol/L)和ERK抑制剂FR180204(15μmol/L)预处理2 h后,再加入不同浓度QUE(0、10、20、40、80和100μmol/L)处理。MTT法检测细胞活力,Transwell法检测细胞迁移,Western blot法检测AKT、内皮型一氧化氮合酶(eNOS)和ERK蛋白表达及磷酸化水平。结果:QUE呈浓度依赖性提高EPCs活力,促进EPCs迁移;PI3K抑制剂BYL719能抑制QUE诱导的EPCs活力和迁移能力,ERK抑制剂FR180204能抑制QUE诱导的EPCs活力,而对EPCs迁移能力并没有影响。QUE能激活AKT、eNOS和ERK蛋白;BYL719能同时抑制AKT和ERK蛋白的激活,FR180204仅能抑制ERK的激活,而未能抑制AKT的激活,但两者对QUE诱导的eNOS蛋白表达均有抑制作用。结论:QUE能部分通过PI3K/AKT/eNOS和ERK/eNOS信号转导通路,提高EPCs活力及迁移能力,促进EPCs发挥心血管保护作用。     

Influence of endogenous and exogenous sex hormones on plasma brain-derived neurotrophic factor

BACKGROUND: Brain-derived neurotrophic factor (BDNF) is a mediator of neuronal plasticity and influences learning, memory and cognitive behaviour. The aim of this study is to assess plasma BDNF variations according to hormonal status. METHODS: A total of 60 subjects were included: 20 fertile ovulatory women, 15 amenorrhoeic women and 25 postmenopausal women. Blood samples were collected after overnight fasting. For 5 out of the 20 fertile women, samples were collected every 2 days throughout the whole menstrual cycle. Following basal evaluation, 10 out of 25 postmenopausal women were administered a hormone replacement therapy (HRT) and reevaluated after 6 months of treatment. Plasma BDNF concentrations were measured by enzyme-linked immunosorbent assay. In fertile women, estradiol (E(2)), progesterone and gonadotrophins were also assessed. RESULTS: In fertile women, luteal phase levels of plasma BDNF were significantly higher than follicular phase levels (P < 0.001). BDNF increased from early follicular phase up to Day 14 of the cycle, reaching a pre-ovulatory peak, similar to E(2). A second rise took place during mid-luteal phase, with a peak on Day 24. Amenorrhoeic subjects, as well as postmenopausal women, showed significantly lower plasma BDNF levels compared with fertile females (P < 0.001). BDNF was positively correlated with E(2) and progesterone and negatively correlated with menopausal age. HRT restored BDNF levels to those present in fertile women during the follicular phase. CONCLUSIONS: Plasma BDNF levels are influenced by hormonal status. Modifications in BDNF circulating levels during the menstrual cycle suggest a potential role for gonadal sex hormones (E(2) and progesterone) in regulating neurotrophin expression.     

Variability in ratings of trustworthiness across the menstrual cycle

This study investigated how trusting behavior varies in naturally cycling women, as a function of sex and attractiveness of players in a trust game, at three distinct phases of the menstrual cycle. Women acted more cautiously in an investment game at the preovulatory phase, compared to the menstrual and the mid-luteal phase. Reduced willingness to trust in strangers was particularly expressed toward male players at this time. The increase of estradiol levels from menses to the preovulatory phase was negatively correlated with trust in attractive male other players, whereas the increase of progesterone levels from menses to the mid-luteal phase was positively associated with trust in unattractive female other players. No particular contribution of a single hormone level could be identified for the generally reduced willingness to trust in strangers in the preovulatory phase. Thus, the results emphasize the impact of the menstrual cycle on interpersonal trust, although the exact mode of hormonal action needs to be further investigated.     

供体年龄影响融合生长内皮祖细胞对平滑肌细胞表型转换及增殖迁移的作用

目的:探讨老龄和年轻个体来源融合生长状态内皮祖细胞(EPCs)对血管平滑肌细胞(SMCs)表型转换以及增殖和迁移的调节作用。方法:脱臼处死1~2月龄、19~26月龄SD大鼠,应用含15%FBS的DMEM/F12培养基(含内皮细胞生长添加剂100 mg/L、肝素100 mg/L、青霉素、链霉素各1×105U/L)培养EPCs,取1~2月龄大鼠腹主动脉,组织块法培养血管SMCs,应用Di I-Ac-LDL与FITC-UEA-1荧光双染以及α-SM-actin免疫荧光分别对EPCs和SMCs进行鉴定。建立细胞共培养体系,上室为融合生长状态的EPCs,下室为SMCs,实验分4组:(1)第3代SMCs(P3)组;(2)第4代SMCs(P4)组;(3)第4代SMCs与年轻大鼠来源EPCs共培养(P4YE)组;(4)第4代SMCs与老龄大鼠来源EPCs共培养(P4AE)组。Western blotting检测α-SM-actin和osteopontin蛋白的表达;[3H]-TdR掺入法检测SMCs增殖;细胞划痕实验检测SMCs的迁移能力。结果:与P3组相比,P4组的SMCsα-SM-actin表达显著下调,而osteopontin表达显著增强;P4YE组SMCs的α-SM-actin及osteopontin表达与P3组比较未见有显著差别;与P4组相比,年轻和老龄大鼠来源的EPCs均显著促进第4代SMCs的α-SM-actin和下调osteopontin的表达,抑制第4代SMCs的增殖和迁移;与老龄大鼠来源的EPCs相比,年轻大鼠来源的EPCs更能够显著延迟SMCs表型由收缩型向合成型转换,抑制SMCs增殖和迁移。结论:共培养融合生长状态的EPCs使血管SMCs表型转换延迟、抑制SMCs增殖和迁移,年轻大鼠来源的EPCs较老龄大鼠来源的EPC更显著延迟血管SMCs表型由收缩型向合成型转换,并具有更强的抑制血管SMCs增殖和迁移的能力。     

Effects of sex, phase of the menstrual cycle and gonadal hormones on pain in healthy humans

Sex differences in pain have been noted; women typically report more pain than men. Gonadal hormones may influence pain reports, and, moreover, such hormones may help to explain sex differences and menstrual cycle differences in pain. This study measured venipuncture and intravenous catherization pain during the follicular and luteal phases of the menstrual cycle in regularly menstruating women. Pain was also assessed in a group of men. Pain ratings were higher in women than men. In women, pain ratings did not differ between the follicular and luteal phases. Estradiol and progesterone increased from follicular to luteal phases. Within-phase analyses revealed that pain ratings were positively correlated with estradiol and progesterone during the luteal phase. Moreover, increases in estradiol and progesterone across the menstrual cycle were positively correlated with increases in pain. These findings suggest that variations in gonadal hormones during the menstrual cycle influence the experience of pain in healthy women.     

There are differences in cerebral activation between females in distinct menstrual phases during viewing of erotic stimuli: a fMRI study

There is evidence that men experience more sexual arousal than women but also that women in mid-luteal phase experience more sexual arousal than women outside this phase. Recently, a few functional brain imaging studies have tackled the issue of gender differences as pertaining to reactions to erotica. The question of whether or not gender differences in reactions to erotica are maintained with women in different phases has not yet been answered from a functional brain imaging perspective. In order to examine this issue, functional MRI was performed in 22 male and 22 female volunteers. Subjects viewed erotic film excerpts alternating with emotionally neutral excerpts in a standard block-design paradigm. Arousal to erotic stimuli was evaluated using standard rating scales after scanning. Two-sample t-test with uncorrected P<0.001 values for a priori determined region of interests involved in processing of erotic stimuli and with corrected P<0.05 revealed gender differences: Comparing women in mid-luteal phase and during their menses, superior activation was revealed for women in mid-luteal phase in the anterior cingulate, left insula, and orbitofrontal cortex. A superior activation for men was found in the left thalamus, the bilateral amygdala, the anterior cingulate, the bilateral orbitofrontal, bilateral parahippocampal, and insular regions, which were maintained at a corrected P in the amygdala, the insula, and thalamus. There were no areas of significant superior activation for women neither in mid-luteal phase nor during their menses. Our results indicate that there are differences between women in the two cycle times in cerebral activity during viewing of erotic stimuli. Furthermore, gender differences with women in mid-luteal phases are similar to those in females outside the mid-luteal phase.     

Cycling time trial performance during different phases of the menstrual cycle

Submaximal exercise performance has not previously been assessed in the late follicular phase of the menstrual cycle, which is associated with a pre-ovulatory surge in oestrogen. Therefore, we compared cycling time trial performance during the early follicular (EF), late follicular (LF) and mid-luteal (ML) phase of the menstrual cycle in trained and untrained eumenorrhoeic women who cycled 30 and 15 km, respectively, in a non-fasted state. The women completed the three cycling time trials on a conventional racing bicycle mounted on an air-braked ergometer. We required resting oestrogen to increase by at least twofold above EF phase values in both the LF and ML phases and this resulted in a number of exclusions reducing the sample size of each group. No significant difference was noted in the finishing time between the different menstrual phases in trained (n=5) or untrained (n=8) group, albeit limited by sample size. However, analysis of the combined trained and untrained group data (n=13) revealed a trend for a faster finishing time (P=0.027) in the LF phase compared to the EF phase as 73% of the subjects showed improvements with an average of 5.2±2.9% (or 2.1±1.1 min) in the LF phase (for =0.05 requires P<0.017). Combined group analysis yielded no difference between performance in the EF and ML phase or between the LF and ML phase. Thus, further research is encouraged to confirm the tendency for a faster time trial in the LF phase, which coincides with the pre-ovulatory surge in oestrogen.     

Alterations in the insulin-like growth factor system during the menstrual cycle in normal women

Objectives: To evaluate the effects of endogenous estrogens and progestins on the IGF-system during the normal menstrual cycle in healthy premenopausal women not using contraceptive drugs. Methods: Nine women had fasting blood samples obtained at 2–3 days intervals during a 5 week study period. Plasma levels of IGF-I, IGF-II, IGFBP-1, IGFBP-3, estradiol and progesterone were measured by radioimmunoassay (RIA) in each sample. IGFBP-3 was also evaluated by Western ligand blot (WLB) and immunoblot. Any differences between the menstrual phase (defined as day 1–5), follicular and luteal phases (separation based on plasma estradiol and progesterone values) were evaluated by the Friedman test. Results: A small but significant difference in plasma levels of IGF-I (P<0.01) and IGFBP-3 (P<0.05) measured by RIA between the three phases were seen with the highest levels found during the follicular phase. No change in plasma levels of IGFBP-1 and IGF-II was found and immunoblots did not reveal any alteration in the ratio of fragmented to intact IGFBP-3 during the menstrual cycle. A positive correlation between plasma levels of IGF-I and estradiol was seen in 8 out of 9 patients (P=0.012). Conclusions: The finding of a slight but significant higher level of plasma IGF-I in the follicular and luteal phases compared with the menstrual phase suggests plasma estradiol may influence the level of this growth factor. This hypothesis is further supported by the finding of a correlation between plasma levels of IGF-I and estradiol but not progesterone in individual patients at different times during the menstrual cycle.     

Endometrial and subendometrial perfusion are impaired in women with unexplained subfertility

BACKGROUND: We used three-dimensional power Doppler angiography (3D-PDA) to examine the periodic changes in endometrial development and subendometrial vascularity during the normal menstrual cycle in 29 women with unexplained subfertility and 19 controls. METHODS: 3D-PDA was performed on alternate days from day 3 of the cycle until ovulation and then every 4 days until menses. VOCAL (Virtual Organ Computer-aided AnaLysis) and shell-imaging were used to define and to quantify the power Doppler signal within the endometrial and subendometrial regions producing indices of their relative vascularity. RESULTS: Women with unexplained subfertility demonstrated significant changes with time (P<0.001) in the indices of vascularity within the endometrium and subendometrium during the menstrual cycle characterized by a pre-ovulatory peak and post-ovulatory fall. These changes mirrored those observed in the control group but were significantly reduced in the endometrium and subendometrium during the mid-late follicular phase and early luteal phase. There were no differences in endometrial thickness or volume between the groups or in the plasma concentrations of estradiol or progesterone. CONCLUSIONS: Endometrial and subendometrial vascularity are significantly reduced in women with unexplained subfertility during the mid-late follicular phase irrespective of estradiol or progesterone concentrations and endometrial morphometry.