宫腔灌注药物在薄型子宫内膜治疗中的应用研究进展

子宫内膜的正常生长对妇女生理周期的顺利进行以及胚胎着床、定位、黏附具有重要意义,如果女性存在子宫内膜薄及月经量减少等问题,容易出现怀孕困难或复发性流产。薄型子宫内膜是指子宫内膜的厚度低于能够获得妊娠的阈值,如何改善薄型子宫内膜患者子宫内膜容受性及妊娠结局是目前生殖医学领域待解决的难点问题。宫腔灌注治疗是一种修复子宫内膜和改善内膜容受性的新方法,宫腔灌注粒细胞集落刺激因子、自体富血小板血浆生长激素在薄型子宫内膜治疗中的应用前景良好,可以改善薄型子宫内膜患者的内膜容受性和妊娠结局。深入研究宫腔灌注在薄型子宫内膜治疗中的应用,可为进一步探索薄型子宫内膜的治疗提供理论依据和研究思路。     

雌激素对雌激素受体阴性的子宫内膜腺癌细胞系JEC的作用

目的探讨不同浓度17β-雌二醇（E2）对雌激素受体（ER）阴性的子宫内膜腺癌细胞系JEC体外生长的影响。方法选用人子宫内膜腺癌细胞系JEC进行体外培养,加入含不同浓度E2的培养液,采用MTT比色、流式细胞术和激光共聚焦扫描显微镜的方法,观察子宫内膜腺癌细胞系JEC细胞的增殖活性、细胞周期时相变化及Ca2＋的变化。结果不同浓度的E2作用JEC细胞1、2、3 d后,其OD值和对照组比较无统计学差异,作用4、5 d后10-7、10-8和10-9mol/L组OD值和对照组比较差异有统计学意义（P〈0.05）,其中10-7mol/L组在第5天OD值与对照组比较有显著差异（P〈0.01）。10-7mol/L E2作用JEC细胞4、5 d后,G0/G1、S＋G2/M期细胞比例有所变化,但与对照组比较无统计学意义（P〉0.05）。10-7mol/L的E2作用JEC 4 d后Ca2＋的平均荧光强度值高于对照组（P〈0.01）。结论 ER阴性的JEC细胞生长可受到雌激素的调控,一定浓度的雌激素对ER阴性JEC细胞生长的调控可能与细胞内Ca2＋增加有关。     

子宫内膜样腺癌组织中TFF3、ER、PR的表达及意义

目的探讨子宫内膜样腺癌（EC）组织中三叶因子3（TFF3）、雌激素受体（ER）及孕激素受体（PR）的表达及意义。方法采用免疫组化sP法检测15例良性子宫肌瘤组织、20例子宫内膜不典型增生组织、42例Ec组织中的TFF3、ER和PR,分析TFF3、ER、PR表达与Ec临床病理参数的关系及其相关性。结果TFF3在良性子宫肌瘤组织中的阳性表达率为20．0％,不典型增生组织中为45．0％,EC组织中为66．7％,不同组织间比较P均〈0．01;TFF3蛋白表达与EC的组织学分级和病理分期有关（P均〈0．05）,与浸润程度和淋巴结转移无关。ER在良性子宫肌瘤组织中的阳性表达率为93．3％,不典型增生组织中为70．O％,EC组织中为54．8％,不同组织间比较P均〈0．05;ER阳性表达与Ec的组织学分级相关（P〈0．05）,与分期、淋巴结转移和浸润程度无关。PR在良性子宫肌瘤组织中阳性表达率为86．7％,不典型增生组织中为75．0％,EC组织中为59．5％,不同组织间比较P〉0．05;PR表达与EC的组织学分级相关（P〈0．05）,与分期、淋巴结转移和浸润程度无关。Ec组织中TFF3表达与ER、PR均呈正相关（r=0．002、0．004,P均〈0．01）。结论Ec组织中TFF3表达明显升高,ER表达明显下降;TFF3、ER、PR对EC的发生、发展及预后可能起重要作用。     

雌激素对子宫内膜细胞凋亡的影响及其机制探讨

目的 观察雌激素对子宫内膜细胞凋亡的影响,并探讨其机制.方法 将原代培养的子宫内膜细胞随机分为C组和E8、E7、E6组.C组不加雌激素,E8、E7、E6组分别加入浓度为10-8、10-7、10-6 mol/L的雌激素.采用流式细胞术检测细胞凋亡率,Real-time PCR法检测前列腺特异性酸性磷酸酶（PSAP）基因;构建PSAP基因干扰载体,并转染至子宫内膜细胞,观察转染前后细胞凋亡率和PSAP基因表达量.结果 C、E8、E7、E6组细胞凋亡率（gate1）分别为10.8％±0.1％、15.5％±0.2％、15.8％±0.1％、16.6％±0.3％,E8组和E7组比较,P＞0.05;其余两两比较,P均＜0.05.C、E8、E7、E6组细胞凋亡率（gate2）分别为4.2％±0.2％、4.5％±0.1％、5.4％±0.2％、5.7％±0.1％,两两比较,P均＜0.05.C、E8、E7、E6组PSAP基因表达量分别为0.46 ±0.05、0.33 ±0.03、0.26±0.03、0.40 ±0.02,两两比较,P均＜0.05.干扰前后细胞凋亡率分别为18.2％±1.3％、10.2％±0.7％,PSAP基因表达量分别为50.4 ±3.2、19.2±1.3,P均＜0.05.结论 雌激素作用后子宫内膜细胞凋亡增加,PSAP基因表达降低;PSAP基因被干扰后,细胞凋亡减少.雌激素可能通过抑制PSAP基因表达促进子宫内膜细胞凋亡.     

层粘连蛋白及其受体67LNR在异位子宫内膜中的表达及临床意义

子宫内膜异位症(EMS)虽属良性病变,但异位内膜细胞具有远处转移和种植能力,这种特性与肿瘤细胞转移侵袭的特性相似。研究证实,细胞黏附分子在子宫内膜的异位种植及生长过程中起重要作用。层粘连蛋白(Laminin,LN)是非胶原糖蛋白的主要成分,广泛分布于基底膜的透明层。1979年首次发现其具有调节细胞分化、生长、黏附和移行等多种功能;分子量为67KD的层粘连蛋白受体(1aminin receptor,LNR)可介导细胞结合于基底膜上的层粘连蛋白,调节细胞的黏附、扩散和迁移。     

重度宫腔粘连反复应用大剂量雌激素致子宫内膜非典型增生1例

患者女,33岁,主因“清宫术后闭经2个月”于2009年1月4日首次入院.2008年因原发不孕行试管婴儿妊娠双胎,孕5个月自然流产,流产后1周行清宫术.术后闭经,有周期性下腹痛.月经史16/3～4/35～40 d,经量中等,无痛经,G1P0.2002年曾行腹腔镜检查发现“盆腔子宫内膜异位症”.其母有胃癌病史.患者体质量指数22.1,妇科内诊无异常.经阴道B超:子宫内膜0.4 cm.宫腔镜检查:宫腔呈窄桶状,宫深5.5 cm,双侧输卵管开口未见,未见正常内膜.于2009年1月6日行宫腔镜宫腔粘连分离术(TCRA).术中打开宫腔,显露双侧输卵管开口,宫壁内膜薄,残余正常内膜面积小于1/3.术后给予戊酸雌二醇(补佳乐)10 mg/d,连续口服2个月余.     

骨髓间充质干细胞对小鼠异位子宫内膜的作用观察

目的观察骨髓间充质干细胞能否迁移至异位子宫内膜并向子宫内膜细胞分化,探讨骨髓间充质干细胞与子宫内膜异位症的关系。方法构建子宫内膜异位症小鼠模型。分离雄性小鼠的骨髓间充质干细胞,用绿色荧光蛋白(GPF)标记后自鼠尾静脉注入模型体内。采用激光共聚焦显微镜观察模型小鼠异位子宫内膜组织中GPF的表达变化,用免疫荧光技术检测异位子宫内膜组织中GPF与角蛋白或波形蛋白复合表达变化。结果注射转染了GPF的骨髓间充质干细胞后第8天,异位子宫内膜碎片中有GPF表达,第20天表达最强,此后逐渐减弱;异位子宫内膜中,无GPF与角蛋白或波形蛋白阳性复合表达。结论骨髓间充质干细胞可迁移至异位子宫内膜组织中,但未发现其能分化为子宫内膜细胞。骨髓间充质干细胞与子宫内膜异位症的关系仍不确定。     

结直肠子宫内膜异位症恶变的研究进展

子宫内膜异位症在育龄期妇女中是一种相对常见的病变,但肠道的子宫内膜异位组织恶变比较罕见。其病因未明,目前用以解释其起源的理论较多。尽管目前针对这种少见疾病无标准治疗方法,但是根治性手术结合术后辅助治疗的模式可作为一种治疗选择。     

子宫内膜增生中PTEN、ER、PR的表达与子宫内膜切除术预后的相关性

目的 探讨子宫内膜增生患者中第10号染色体同源丢失性磷酸酶-张力蛋白基因(PTEN)、雌激素受体(ER)、孕激素受体(PR)的表达与宫腔镜子宫内膜切除术(TCRE)预后的相关性.方法 采用免疫组化法对78例因功能失调性子宫出血行TCRE患者的子宫内膜行PTEN、ER、PR测定,其中增殖期内膜15例、单纯性增生内膜27例、复杂性增生内膜17例、不典型增生内膜19例.结果 PTEN、ER、PR在增殖期、单纯性增生、复杂性增生及不典型增生内膜中均有阳性表达,其阳性表达均呈递减趋势.等级相关分析结果显示PTEN、ER、PR表达异常与子宫内膜组织学分级均呈负相关.PTEN、ER、PR阳性表达的不同与TCRE预后有关,随着PTEN、ER、PR阳性表达强度的逐渐降低,TCRE术后复发率呈递增趋势.结论 PTEN、ER、PR与子宫内膜病变的发生发展密切相关,PTEN、 ER、PR的表达减低与TCRE预后相关.     

不同应激方式对子宫内膜异位症子宫内膜细胞凋亡影响的研究进展

子宫内膜异位症是一种雌激素依赖的女性慢性炎症性疾病,可导致盆腔疼痛、不孕及卵巢癌易感性等。子宫内膜异位症的异位病变具有选择性生存优势,可抑制子宫内膜细胞的凋亡能力;而低氧应激、内质网应激、线粒体应激及细胞内激酶的级联激活对于细胞凋亡有重要作用,不同的应激通过不同方式影响细胞凋亡,进而影响子宫内膜异位症的发生发展。     

精子DNA损伤在辅助生殖中的研究进展

男性生育力受到许多因素的影响,其中精子DNA损伤一直是生殖医学领域研究的热点之一。该损伤可由多种因素诱发精子DNA双链断裂或使其双链变性而形成。该文通过分析精子生成错误、氧化应激、精子染色质包装异常、疾病等因素对精子DNA损伤的影响,阐述精子DNA损伤检测在辅助生殖医学中的研究现状,为进一步阐明精子DNA损伤的发生机制以及如何为辅助生殖技术选用高质量精子提供基础材料。     

Epigenetics and assisted reproductive technology

During gametogenesis, the female and male germ cells undergo a process whereby imprinting marks are erased from the genome. During the later stages of germ-cell development, the methylation marks of the female and male germ lines are re-established. A second phase of demethylation of the genome occurs at the time of fertilization, and during development of the early embryo. Assisted reproductive technology involves several steps that subject the gametes and early developing embryos to environmental stress, and this is the primary reason for an increased interest in the putative link between these techniques and imprinting disorders. Although animal studies support a link between assisted reproductive techniques (ARTs) and imprinting disorders, via altered methylation patterns, data in humans are inconsistent. Here we provide an overview of the field of epigenetics in relation to ARTs.     

Epigenetics, genomic imprinting and assisted reproductive technology

Epigenetic mechanisms play a key role in regulating gene expression. One hallmark of these modifications is DNA methylation at cytosine residues of CpG dinucleotides in gene promoters, transposons and imprinting control regions. Genomic imprinting refers to an epigenetic marking of genes that results in monoallelic expression depending on their parental origin. There are two critical time periods in epigenetic reprogramming: gametogenesis and early preimplantation development. Major reprogramming takes place in primordial germ cells in which parental imprints are erased and totipotency is restored [1]. Imprint marks are then and re-established during spermatogenesis or oogenesis, depending on sex [1], [2] and [3]. Upon fertilization, genome-wide demethylation occurs followed by a wave of de novo methylation, both of which are resisted by imprinted loci [4]. Epigenetic patterns are usually faithfully maintained during development. However, this maintenance sometimes fails, resulting in the disturbance of epigenetic patterns and human disorders. For example, two fetal growth disorders, the Beckwith-Wiedemann (BWS) and the Silver-Russell (SRS) syndromes with opposite phenotypes, are caused by abnormal DNA methylation at the 11p15 imprinted locus [5], [6] and [7]: respectively loss of methylation at the Imprinting Region Center (ICR2) or gain of methylation at ICR1 in BWS and loss of methylation at ICR1 in SRS. Early embryogenesis is a critical time for epigenetic regulation, and this process is sensitive to environmental factors. The use of assisted reproductive technology (ART) has been shown to induce epigenetic alterations and to affect fetal growth and development [8], [9], [10] and [11]. In humans, several imprinting disorders, including BWS, occur at significantly higher frequencies in children conceived with the use of ART than in children conceived spontaneously [12] and [13]. The cause of these epigenetic imprinting disorders (following ART, unfertility causes, hormonal hyperstimulation, in vitro fertilization-IVF, Intracytoplasmic sperm injection-ICSI, micro-manipulation of gametes, exposure to culture medium, in vitro ovocyte maturation, time of transfer) remains unclear. However, recent data have shown that in patients with BWS or SRS, including those born following the use of ART, the DNA methylation defect involves imprinted loci other than 11p15 [14] and [15] (11p15 region: CTCF binding sites at ICR1, H19 and IGF2 DMRs, KCNQ1OT1 [ICR2], SNRPN [chromosome 15 q11-13], PEG/MEST1 [chromosome 7q31], IGF type2 receptor and ZAC1 [chromosome 6q26 et 6q24 respectively], DLK1/GTL2-IG-DMR [chromosome 14q32] and GNAS locus [chromosome 20q13.3]). This suggests that unfaithful maintenance of DNA methylation marks following fertilization involves the dysregulation of a trans-acting regulatory factor that could be altered by ART.     

Studies on sperm chromosomes in patients with severe male factor infertility undergoing assisted reproductive technology treatment

The aim of the study was to determine the rate of chromosome abnormalities in testicular sperm after intracytoplasmic sperm injection due to severe male factor infertility. The study groups included patient with non-obstructive azoospermia (n=9), obstructive azoospermia (n=10), Klinefelter's syndrome (n=5) and normal controls (n=6, groups I-VI, respectively). The mean serum levels of FSH 17.5+/-8.2 (P<0.05), 3.5+/-2.6, 29.8+/-13.0 (P<0.05) and 3.1+/-0.4 mIU/ml, respectively. The rates of chromosome abnormalities were 19.6% (P<0.001), 8.2% (P<0.001), 6.3 and 1.6%, respectively. Chromosomes X and Y were significantly more involved in the aneuploidy than chromosome 18 in groups I and II. The present findings demonstrate a linkage between gonadal failure (high serum FSH levels) and sperm chromosome abnormalities. Our findings may explain the increased incidence of perinatal sex chromosome abnormalities found in severe male factor patients. Patients with non-mosaic Klinefelter's syndrome have comparable risk for sex chromosomes aneuploidy as the rest of the patients with azoospermia. Therefore, genetic screening during pregnancy or before embryo replacement should be carefully considered in severe male factor patient following in vitro fertilization (IVF).     

精子质量对不孕不育患者辅助生殖技术选择的指导作用

目的探讨精子质量对不孕不育患者辅助生殖技术选择的指导作用。方法选择201对行常规体外受精胚胎移植术(IVF-ET)的不孕不育夫妇,其中行IVF-ET成功(二极体的胚胎数量占胚胎总数≥30%)者126对(IVF-ET组),IVF-ET失败(二极体的胚胎数量占总胚胎数<30%)后行补救卵胞浆内单精子注射术(R-ICSI)75对(R-ICSI组)。比较两组精液常规、精子DNA完整率、精子顶体完整率、顶体反应率;体外受精后比较两组受精率、优胚胎率。结果 IVF-ET组精子密度、活力及正常形态率均高于R-ICSI组(P均<0.05);R-ICSI组精子DNA断裂指数(DFI)明显高于IVF-ET组(P<0.05),顶体完整率、顶体反应率两组比较无统计学差异(P均>0.05);R-ICSI组受精率、优胚率低于IVF-ET组(P均<0.05)。结论当精子密度低于11.1×106/m L,正常形态率低于1.4%,DFI高于15.9%,顶体完整率低于77.3%,顶体反应率低于15.7%时建议直接行R-ICSI助孕。     

Gonadotropins as pharmacological agents in assisted reproductive technology and polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is a complex endocrinopathy associated with subfertility/infertility and pregnancy complications. Most PCOS women opt for assisted reproductive technologies (ART) for successful conception; however, optimization of the relative doses of the gonadotropins [follicle-stimulating hormone (FSH), luteinizing hormone (LH)/human chorionic gonadotropin (hCG)] for appropriate steroidogenesis, without causing ovarian hyperstimulatory syndrome (OHSS), is challenging. Embryonic factors probably do not contribute to pregnancy loss in PCOS women, albeit hormonal imbalance impairs the metabolic microenvironment critical for oocyte maturation and endometrial receptivity. Certain clinical studies have confirmed the role of metabolic corrections in increasing the rate of pregnancy in PCOS women. This review focuses on the impact of untimely high LHCGR and/or LH levels on oocyte/embryo quality, pregnancy outcomes in ART, and exploring LHCGR as a potential drug target in PCOS women.     

Rare congenital disorders,imprinted genes,and assisted reproductive technology

CONTEXT: During the past two decades, assisted reproductive technologies (ARTs) have revolutionised the treatment of infertility. ARTs now account for between 1% and 3% of annual births in many western countries and in-vitro fertilisation (IVF) services are growing worldwide. In general, the incidence of abnormalities at birth is reassuringly low and children develop normally. Nevertheless, it is important to monitor the safety of ARTs as clinical protocols evolve and new technologies emerge. STARTING POINT: Three recent studies all report an unexpectedly high incidence of Beckwith-Wiedemann syndrome (BWS) in children conceived with ARTs. Six of 149 cases were reported from a British BWS registry (J Med Genet 2003; 40: 62-64); the same numbers were recorded in a French registry (Am J Hum Genet 2003; 72: 1338-41), and a further seven children have been reported in the USA (Am J Hum Genet 2003; 72: 156-60). These frequencies are extraordinarily high for such a rare congenital condition and such findings are reminiscent of reports of sporadic cases of the imprinting disorder, Angelman syndrome, which has also been linked with ARTs. WHERE NEXT? Continuing surveillance of children conceived with ARTs is needed, including monitoring birth defects, development, and cancer. Studies will need to be prospective and multicentre, and should include molecular characterisation of epigenetic abnormalities, including the methylation status of imprinting control regions within imprinted gene clusters.     

Recent advances in assisted reproductive technologies

Assisted reproductive technologies (ART) have received considerable attention, both clinically and empirically. Drs. Steptoe and Edwards removed one oocyte surgically from a woman with infertility related to tubal disease. They fertilized this oocyte in vitro and transferred the formed embryo to the woman’s uterus and achieved pregnancy and delivery. The technique of in vitro fertilization (IVF) and embryo transfer (ET) quickly became widely utilized for other causes of infertility as well as for tubal disease. In the last 5 years there has been a number of new developments that are reviewed in this article. The most important and now widely practiced technology has been direct intracytoplasmic injection (ICSI) of the husband’s sperm into the wife’s oocyte. This was developed for treatment of infertility related to low sperm count. Subsequently it was shown that sperm can be aspirated from epididymis or found in testicular biopsy in obstructive azoospermia. Another promising development is in vitro maturation (IVM) of immature oocytes. This has the potential of avoiding ovarian hyperstimulation, which can be uncomfortable and occasionally dangerous. Some oocytes are unable to fertilize and/or develop into normal embryos. It may be possible that the problem is with the machinery of cytoplasm of the oocyte. Therefore cytoplasmic transfer from a normal oocyte to an abnormal oocyte may overcome the problem. Infertile couples may be faced with many psychological problems that become even more complex with various treatments. Whereas donation of oocytes or embryos can be technically quite simple, there are many psychological issues involved. As can be gathered from aforementioned discussions, the treatments developed for infertility appear to be somewhat illogical and in the style of “shot gun therapy.” In the field of infertility, as in other areas of medicine, it is of paramount importance to know the details of disease mechanisms. This in turn will allow specific and logical treatments to be developed.     

诱导痰技术在慢性阻塞性肺疾病诊治中应用的研究进展

慢性阻塞性肺疾病(COPD)是一种包括慢性支气管炎及肺气肿的慢性气道非特异性炎症。诱导痰技术是一种无创性气道采样方法,通过超声雾化吸入高渗盐水诱导痰液生成,从而进一步分析其中的细胞成分和上清液中的可溶性介质,以评估呼吸道炎症的性质和严重程度。近年来,诱导痰技术越来越多地应用于无痰、少痰的COPD患者中,可用于评估患者的气道炎症及气流阻塞严重程度、指导激素的使用、评估疗效及预后、区分COPD表型等。