氯菧酚胺对增生期子宫内膜组织学的影响及其与血清雌激素的关系

本文应用图像分析仪定量分析２１例不孕妇女氯菧酚胺（ＣＣ）用药前后周期第１２天子宫内膜组织结构，及其与血Ｅ２浓度的关系。结果表明：ＣＣ周期血Ｅ２显著高于自然周期同期水平，分别为１９５１．９２±１２４９．２８ｐｍｏｌ／Ｌ和４４９．６９±２５７．３６ｐｍｏｌ／Ｌ（Ｐ＜０．００１）；腺体面积小于自然周期值，分别为５２３５．５３±１００４．５２μｍ２和６４４４．６１±１２７３．２３μｍ２（Ｐ＜０．０５）。相关分析示自然周期血Ｅ２浓度与腺体面积呈正相关性（Ｐ＝０．００２），ＣＣ周期两者无相关性（Ｐ＞０．０５）。提示：ＣＣ阻断Ｅ２作用，抑制增生期子宫内膜腺体发育，但与高血Ｅ２不相关。     

氯菧酚胺对增生期于宫内膜的影响及其与血清雌激素的关系

应用阴道超声观测１５例不孕妇女氯酚胺（ＣＣ）应用前后增生期子宫内膜的厚度，放射免疫法测量血清雌二醇（Ｅ２）浓度。结果表明：ＣＣ周期第１２天血Ｅ２浓度显著高于自然周期同期水平（Ｐ＜０．００１），而子宫内膜厚度显著较自然周期同期厚度为薄（Ｐ＜０．０５），相关分析示自然周期血巳浓度与子宫内膜厚度呈正相关性．而ＣＣ周期两者无相关性。提示：ＣＣ阻断Ｅ２对子宫内膜的作用，从而影响增生期子宫膜内增厚，内膜厚度与高血Ｅ２浓度不相关。     

克罗米酚对子宫内膜雌,孕激素受体的影响

目的：观察克罗米酚（ＣＣ）对子宫内膜不同类型细胞内雌激素受体（ＥＲ）和孕激素受体（ＰＲ）影响。方法：采用抗ＥＲ、ＰＲ单克隆抗体的链霉菌抗生物素－过氧化酶连结法的免疫组化法，对１６在不明原因９１１例）或男性因素（５例）不孕妇女的自然周期和用药周期的增生晚期和分泌晚期子宫内膜细胞内ＥＲ、ＰＲ，进行定位和半定量分析。结果；用药周期中，增生晚期和分泌晚期的内膜上此细胞、间质细胞明显低下（Ｐ〈０．０５），在     

上皮性卵巢癌DNA含量、细胞核形态及其对患者预后价值的探讨

目的：探讨ＤＮＡ含量、核形态参数对预测上皮性卵巢癌患者预后的价值。方法：应用计算机图像分析系统测定原发性上皮性卵巢癌３２例的细胞核ＤＮＡ含量、倍体水平、面积、周长及形状因子５个参数。结果：卵巢癌临床晚期及转移病例的ＤＮＡ含量明显增高，与早期及无转移组相比，差异均有显著性（Ｐ＜０．０２５）。６３％为异倍体肿瘤，３７％为二倍体肿瘤。组织学Ⅰ、Ⅱ、Ⅲ级的异倍体率分别为３８％、５７％、９０％，呈递增趋势。Ⅰ级与Ⅲ级相比，差异有显著性（Ｐ＜０．０５）。腹水阳性组及转移组的异倍体率明显增加，分别为７３％及７６％。ＰＣＮＡ阳性组及阴性组的核形状因子差异有显著性（Ｐ＜０．０５）。核面积和周长在诸预后因素间差异无显著性（Ｐ＞０．０５）。结论：细胞核ＤＮＡ含量和倍体水平是影响卵巢癌患者预后的重要因素；核形状因子亦可作为评估肿瘤生物学行为的一项指标。     

hMG对子宫内膜、卵泡发育及白细胞雌激素受体作用的探讨

目的：探讨ｈＭＧ对子宫内膜、卵泡发育及白血胞雌激素受体含量的变化及其临床意义。方法：应用阴道超声、放免法和放射配体结合法对１８例自然周期和１６例ｈＭＧ促排卵周期的不孕症患者子宫内膜厚度、分型、成熟卵泡数、血浆雌二醇及孕酮浓度和外周血白细胞雌激素受体含量进行测定比较。结果：①ｈＭＧ组围排卵期子宫内膜厚度大于自然周期组（Ｐ＜０．０５）;成熟卵泡数多于自然周期组（Ｐ＜０．０５）;子宫内膜Ａ、Ｂ型回声比例高于自然周期组（Ｐ＜０．０５）;②ｈＭＧ组围排卵期血浆雌二醇、孕酮浓度明显高于自然周期组（Ｐ＜０．０５）;③ｈＭＧ组围排卵期白细胞雌激素受体含量较自然周期组明显升高（Ｐ＜０．０１）。结论：①ｈＭＧ可使促排卵周期成熟卵泡数增加,子宫内膜分型改善,血浆Ｅ２浓度及白细胞雌激素受体含量升高,有改善子宫内膜容受性的作用;②ｈＭＧ可能有致卵泡过早黄素化的作用;③测定白细胞雌激素受体可间接反映子宫内膜雌激素受体的变化,具有无创、可靠、重复性好的优点     

应用图象分析测定卵巢囊腺癌细胞核DNA含量及形态参数

应用图象分析技术对５６例卵巢囊腺癌、１２例良性囊腺瘤和１０例正常卵巢上皮组织的细胞核ＤＮＡ含量及形态参数进行定量研究。结果表明：囊腺癌细胞核ＤＮＡ含量比囊腺瘤和正常卵巢均高，ＤＮＡ直方图图形分布弥散，出现多倍体细胞；囊腺癌组细胞的核形状因子与其它两组比较，差异有非常显著意义，细胞核周长，核面积差异无显著意义。     

应用粘附细胞激光定量分析仪测定子宫内膜细胞脱氧核糖核酸含量

应用粘附细胞激光定量分析仪测定子宫内膜细胞脱氧核糖核酸含量王巧玲金志魁粘附细胞激光定量分析仪（ＡＣＡＳ５７０），是目前用于细胞学研究的较先进的一种仪器。它可直接对活细胞进行观察。本研究选用ＡＣＡＳ５７０对子宫内膜粘附细胞ＤＮＡ含量进行测定，以探讨子宫...     

分泌期及早期子宫内膜巨噬细胞及自然杀伤细胞的测定

目的 探讨子宫内膜局部免疫微环境对妊娠的影响。方法 对１９例孕６￣９周的正常妊娠和１１例正常分泌期妇女,采用流式细胞技术检测蜕膜及内膜的巨噬细胞及自然杀伤（ＮＫ）细胞。结果 蜕膜组织中巨噬细胞（白细胞分化抗原分化簇（ＣＤ）１４阳性即ＣＤ１４」及ＮＫ细胞（ＣＤ５６阳性即ＣＤ５６）含量较分泌期子宫内膜组织增加,差异有极显著性。ＮＫ细胞的ＣＤ５６ＣＤ１６亚群含量增加,差异有极显著性,而ＣＤ５６ＣＤ１６亚     

离体异位子宫内膜细胞中单核细胞趋化蛋白-1的测定

目的　探讨异位子宫内膜细胞产生单核细胞趋化蛋白１（ＭＣＰ１）的临床意义。方法　收集１５ 例子宫内膜异位症患者的异位内膜组织进行体外细胞培养,分别加入纯培养液（ 对照组） 、含白细胞介素１β（ＩＬ１β）２ ｎｇ／ｍｌ（ＩＬ１β诱导组）及肿瘤坏死因子α（ＴＮＦα）２０ ｍｇ／ｍｌ（ＴＮＦα诱导组）的培养液培养４ 小时,应用斑点杂交、免疫组织化学及夹心酶联免疫吸附试验检测各组异位细胞中ＭＣＰ１ｍＲＮＡ蛋白的表达水平及其培养上清中ＭＣＰ１ 的含量。结果　与对照组相比,异位内膜细胞经ＩＬ１β及ＴＮＦα作用后表达的ＭＣＰ１ ｍＲＮＡ和蛋白水平显著增加（ Ｐ＜ ０．０１ ,Ｐ＜０．０５）;培养上清中ＭＣＰ１ 的含量也显著增加（Ｐ＜０ ．０１ ,Ｐ＜ ０．０５）。结论　ＩＬ１β及ＴＮＦα可刺激异位内膜细胞中ＭＣＰ１ 的表达显著增加,这可能和子宫内膜异位症的病变发展有关。     

子宫的内分泌功能及作用

子宫不仅是一个受激素作用的靶器官，也是一个功能复杂的内分泌器官。子宫的内分泌功能主要由子宫内膜细胞产生。内膜细胞由腺上皮细胞（ｅｎｄｏｍｅｔｒｉａｌｅｐｉｔｈｅｌｉａｌｃｅｌｓ，ＥＥＣ）及间质细胞（ｅｎｄｏｍｅｔｒｉａｌｓｔｒｏｍａｌｃｅｌｓ，Ｅ...     

月经周期中不同类型人子宫内膜体外培养的细胞形态学特点

目的:探讨不同类型子宫内膜细胞体外培养的细胞形态学特点。方法:在月经周期的不同时期,通过B超检测判断患者子宫内膜的厚度和类型,取不同形态分型的人子宫内膜进行体外培养,倒置显微镜下观察子宫内膜体外培养的细胞形态学特性,并通过免疫荧光法进行鉴定,采用培养板单层贴壁细胞的原位计数方法,计算子宫内膜间质细胞和腺上皮细胞数量及比例。结果:不同类型子宫内膜经培养后,其中间质细胞形态呈扁平状,胞质透明,核圆、居中,波形蛋白表达阳性;腺上皮细胞呈多角形或蝌蚪形,旋涡状排列,核圆而大,角蛋白表达阳性。A型子宫内膜组织腺细胞贴壁率为85.8±5.7%,贴壁培养后腺上皮细胞的比例占43.3±11.7%,间质细胞比例占56.7±11.7%;B型子宫内膜组织腺细胞贴壁率为8.7±3.9%,贴壁培养后腺上皮细胞的比例占13.3±3.3%,间质细胞比例占86.7±3.3%;C型子宫内膜组织腺细胞贴壁率为6.0±1.9%,贴壁培养后腺上皮细胞的比例占12.3±3.0%,间质细胞比例占87.7±3.0%。A组与B组、C组两组腺上皮细胞贴壁率及贴壁后腺细胞比例有明显统计学差异(P0.05),B、C组之间腺上皮细胞贴壁率及腺细胞比例无明显统计学差异(P0.05)。结论:B、C型子宫内膜体外培养腺上皮细胞贴壁比例低于间质细胞,体外培养后以间质细胞为主;A型子宫内膜细胞培养所得的间质细胞及腺细胞均较多,腺细胞贴壁率及腺细胞比例明显高于B、C型子宫内膜,是用于胚胎着床机制等实验研究较好的标本来源。     

改良筛网法分离培养人子宫内膜细胞

目的:探索一种能获得较高纯度和较多数量子宫内膜细胞的分离培养方法。方法:取12例患者各1mg新鲜子宫内膜组织,经0.25%Ⅰ型胶原酶(含有0.005%DNA酶)消化后,将所得细胞悬液一分为二,分别采用传统筛网法和改良筛网法(二次筛网过滤和低速离心相结合)分离并记录所得细胞数;体外培养后采用光学显微镜观察,用免疫细胞化学和免疫荧光化学方法鉴定纯度,比较两种方法分离子宫内膜细胞的纯度和数量。结果:两种方法均有11例组织分离培养成功。传统筛网法分离的子宫内膜腺上皮细胞和间质细胞的纯度分别为(89.54±3.75)%和(92.82±1.60)%,细胞数分别为(0.47±0.09)×105个和(23.73±5.37)×105个。改良筛网法分离的腺上皮细胞和间质细胞纯度分别为(91.95±0.47)%和(99.28±0.21)%,细胞数分别为(8.09±1.24)×105个和(27.23±0.50)×105个。改良筛网法分离的腺上皮细胞数量和间质细胞纯度均明显高于传统筛网法,有统计学差异(P均=0.000)。结论:改良筛网法操作简单,能获得较高纯度的子宫内膜间质细胞及较多的子宫内膜腺上皮细胞。     

子宫内膜腺上皮及基质细胞分离、培养作为子宫内膜异位症体外细胞模型的探索

目的 :探索在位子宫内膜腺上皮及基质细胞的分离、培养作为子宫内膜异位症 (EMs)的体外细胞模型。方法 :通过酶解、系列过滤、沉降及贴壁纯化等技术 ,分离、纯化及培养 15份EMs患者的在位子宫内膜腺上皮细胞及其基质细胞 ,并拟传代。结果 :11份标本获得成功 ;每 1g新鲜子宫内膜组织可得到 (8～ 12 )× 10 6个原代基质细胞及 (4～ 8)× 10 6个原代腺上皮细胞 ;基质细胞纯度率可达 95 %以上 ,腺上皮细胞的纯度率约为90 %。基质细胞可以有限的传代 ,腺上皮细胞不能传代。通过改良步骤 ,可提高基质细胞的产量。结论 :在位子宫内膜腺上皮及其基质细胞的分离、培养可作为EMs的体外细胞模型之一。     

左旋18-甲基炔诺酮和米非司酮对分泌中期人子宫内膜HOXA11基因表达的影响

目的:通过口服孕激素类(左旋18-甲基炔诺酮,Levenorgestrel,LNG)和抗孕激素类(米非司酮,RU486)避孕药,研究孕激素对分泌中期人子宫内膜HOXA11基因表达的影响。方法:30名自愿者,于正常月经周期排卵后d7,刮取内膜组织做自身对照;实验周期在排卵前或排卵后2d,口服小剂量LNG(18例)或RU486(12例),同样在排卵后d7刮取内膜组织。用原位杂交方法检测服药子宫内膜HOXA11基因表达的变化。结果:对照组(分泌中期)内膜,间质细胞普遍表达HOXA11mRNA,而腺上皮HOXA11的表达则呈弱阳性或阴性。口服LNG后,子宫内膜形态变化不明显;内膜腺上皮HOXA11表达量进一步下降或无明显变化(即用药前后均表达阴性);间质细胞HOXA11表达增强。服用RU486后,内膜发育明显延迟,腺上皮HOXA11表达强度普遍大于对照组,而间质细胞的表达变化无明显规律。结论:孕激素对内膜腺上皮HOXA11基因的表达起负调控作用;正常分泌中期,内膜腺上皮HOXA11表达减弱或消失是内膜分化成熟的标志。     

Endometrial hormone receptors and proliferation index in the periovulatory phase of stimulated embryo transfer cycles in comparison with natural cycles and relation to clinical pregnancy outcome

OBJECTIVE: To investigate the endometrial steroid receptors and proliferation index in GnRH analogue/hMG-stimulated cycles in comparison with natural cycles and their relation to clinical pregnancy outcome. DESIGN: Prospective observational study. SETTING: Tertiary referral center. PATIENT(S): Twenty-seven stimulated patients with GnRH agonist and hMG. Twenty normo-ovulatory patients were the natural cycle controls. INTERVENTION(S): Endometrial aspiration biopsies: in stimulated cycles on the day of oocyte retrieval within the ET cycle (Day OPU) (n = 20) or 2 days later (Day OPU + 2) (n = 7); in natural cycles on the natural day of ovulation (Day NO) (n = 10) or on the day of ovulation + 2 (Day NO + 2) (n = 10). MAIN OUTCOME MEASURE(S): Comparison of endometrial maturation, estrogen (ER) and P receptor (PR), and proliferation index by immunohistochemistry in natural and stimulated cycles, correlation with pregnancy outcome in stimulated cycles. RESULT(S): Stimulated cycles Day OPU showed significantly advanced endometrial maturation compared to natural cycles Day NO; stromal ER and glandular and stromal PR staining was lower in stimulated than in natural cycles, but higher on Day OPU than on Day NO + 2; proliferation index was lower in all stimulated cycles. Steroid receptors and proliferation index in stimulated cycles were unrelated to clinical pregnancy occurrence. CONCLUSION(S): Compared to natural cycles, ovarian stimulation induced an imbalance in endometrial ER and PR and led to a profound antimitotic effect in the peri-ovulatory phase. These parameters were, however, not predictive of clinical pregnancy in cycles with ET.     

Normal human endometrium. An ultrastructural survey

An ultrastructural review of the cyclic changes of endometrial surface and glandular epithelial cells, stromal cells and the stromal microvasculature is presented. Endometrial biopsies were collected from infertile patients with endometriosis or tubal dysfunction. During the proliferative phase, the ergastoplasm and Golgi apparatus of surface and glandular epithelial cells become well-developed, while increasing numbers of mitochondria are seen. In the early secretory phase, the postovulatory triad composed of glycogen accumulation, nuclear channel system and giant mitochondria is described and its functional significance is discussed. Premenstrual cellular involution due to enhanced lysosomal activity such as apoptosis and lysosomal wrapping is detailed. Before ovulation, the endometrial stromal cells are primarily involved in the secretion and remodelling of the extracellular connective tissue fibers and ground substance. After predecidual change, the stromal cells exhibit marked macropinocytotic and phagocytotic properties. We suggest that these proteolytic activities of the stromal cells, together with the presence of blood-derived phagocytes, may account for endometrial clearance, since there are no lymphatic channels in the superficial endometrium. According to this hypothesis, tissue fluid may be drained via the fenestrated (venous) endometrial blood capillaries.     

Investigations on isolation, purification and cultivation of human endometrial cells and on the in vitro inhibin expression in glandular epithelial cells

The separate in vitro cultivation of isolated and purified human endometrial glands and stromal cells seems to be the most attractive experimental way of studying the endometrial function on cellular level. In this paper a new method has been described to establish monolayer cultures of isolated endometrial stromal and epithelial cell populations. After a first collagenase digestion, stromal and epithelial cells were separated by filtration. The glandular epithelial cells were further purified with two collagenase digestion steps, filtration, a differential sedimentation at unity gravity and a Ficoll gradient centrifugation. Stromal cells were isolated with the use of erylyse-buffer, filtration and differential sedimentation at unity gravity. A significant higher inhibin production was observed during late secretory compared to proliferative and early secretory phase. Therefore, glandular epithelial cells maintain in vitro their initial differentiation. The higher inhibin concentration during secretory phase implicates a substantial role in endometrial function and maturation. Therefore, inhibin could be used as a marker of endometrial differentiation. Experiments on isolated glandular epithelial cells should be performed within two weeks. The method described here allows the propagation in vitro of separate endometrium cell types which can be used to study endometrial function as well as implantation mechanisms.     

月经周期人子宫内膜腺上皮和基质细胞HOXA11表达的差异

目的:探讨HOXA11在月经周期人子宫内膜腺上皮和基质细胞中的表达规律及其生理意义。方法:免疫组织化学方法观察38例子宫内膜腺上皮和基质细胞HOXA11蛋白质的表达;用细胞分离筛选法,分离出子宫内膜腺上皮细胞和基质细胞,采用半定量RT-PCR和Dot-blot方法分别观察HOXA11在腺上皮和基质细胞中的表达。结果:腺上皮细胞HOXA11在分泌中晚期表达量较增生期和分泌早期显著降低;基质细胞HOXA11的表达从增生期到分泌期逐渐增加,以分泌中晚期表达量最高。结论:内膜腺上皮和基质细胞HOXA11表达在分泌中晚期变化最显著,而且其表达量呈反相变化,即腺上皮表达量下降,基质细胞表达量增加。提示HOXA11基因与着床期子宫内膜的分化、成熟密切相关;其对内膜腺上皮和基质细胞的调控机制可能不尽相同。     

Differential expression of endometrial integrins and progesterone receptor during the window of implantation in normo-ovulatory women treated with clomiphene citrate

OBJECTIVE: To assess the effect of clomiphene citrate (CC) on endometrial epithelial integrins and P receptors (PR) during the window of implantation. DESIGN: Controlled, prospective, clinical study. SETTING: Teaching hospital and university research laboratory. PATIENT(S): Thirty-one fertile, normo-ovulatory women participated in this trial. Thirteen women exhibited a CC-stimulated cycle with 50 mg on days 5-9, and 18 women with spontaneous menstrual cycles served as controls. INTERVENTION(S): Endometrial biopsies in the midluteal phase. MAIN OUTCOME MEASURE(S): Immunohistochemical determination and endometrial cellular localization of alpha1, alpha v, beta3, and alpha4 epithelial integrins and PR during the window of implantation. The staining intensity was assessed by a semiquantitative index (HSCORE) and compared by nonparametric Mann-Whitney test. RESULT(S): Higher plasma levels of P and E2 and delayed histologic dating of the endometrium (38%) were features of CC-treated women. In addition, a low epithelial beta3 integrin expression and persistent PR were observed in glandular epithelial cells of "out-of-phase" endometrial biopsies from CC-treated women. In contrast, in "in-phase" biopsies, neither epithelial PR nor beta3 integrin were different from spontaneous control cycles. There was no difference in the expression of alpha1, alpha v, and alpha4 between the groups studied. CONCLUSION(S): The administration of clomiphene produces aberrant endometrial beta3 integrin expression in conjunction with a failure in the down-regulation of PR during the window of implantation in a significant number of normo-ovulatory women, notwithstanding the higher plasma P levels. Therefore, CC might affect the expression of endometrial receptivity markers.     

Apoptosis and differential expression of apoptosis-related proteins in endometriotic glandular and stromal cells

OBJECTIVE: Apoptosis is an important regulator of eutopic endometrial function. Endometriosis, the growth of endometrial tissue outside the uterus, could result from increased cellular proliferation or decreased apoptosis in response to appropriate stimuli. The objective of this study was to evaluate the rate of apoptosis and the expression of apoptosis-related Bcl-2 and Bax proteins in endometriotic tissues within the glandular and stromal compartments, according to the phase of the menstrual cycle and the stage of disease. METHODS: Ovarian endometriosis samples were evaluated in 75 women who had surgery at a university hospital. Apoptotic cells were detected with the use of the dUTP nick-end labeling (TUNEL) assay. Bcl-2 and Bax expression were assessed by immunohistochemical techniques. RESULTS: The percentage of apoptotic cells was significantly higher in endometriotic stromal cells (73.3%) compared with glandular cells (48%; P =.002). In contrast, the expression of the apoptosis-related proteins Bcl-2 and Bax was significantly lower in the endometriotic stroma (17.3% for both) than in the glandular epithelium (38.6% and 41.3%, respectively; P <.004). No significant menstrual cycle phase-dependent changes or endometriosis stage-related changes were observed in TUNEL, Bcl-2, or Bax positivity within ovarian endometriotic tissues. CONCLUSION: Apoptosis occurs in ovarian endometriotic lesions at significantly higher levels in the stroma than the glandular epithelium. However, Bcl-2 and Bax proteins are distributed preferentially in glandular epithelial cells. The apoptotic rate as well as Bcl-2 and Bax expression in ovarian endometriotic cells were not affected by the stage of endometriosis or the phase of the menstrual cycle.