乙酰肝素酶基因在子宫内膜异位症患者在位与异位子宫内膜组织中的表达

目的　研究乙酰肝素酶基因在子宫内膜异位症发病中的作用。方法　应用原位杂交技术 ,检测腹腔镜手术中证实为子宫内膜异位症患者 (EM组 ,2 3例 )的在位和异位子宫内膜组织乙酰肝素酶mRNA的表达 ,并与正常妇女 (对照组 ,2 5例 )子宫内膜组织比较。结果　 ( 1)EM组在位与异位子宫内膜乙酰肝素酶mRNA阳性表达率一致 ,但异位子宫内膜组织中 ,阳性细胞着色较深。在位子宫内膜腺上皮细胞较间质细胞胞浆染色深 ,异位子宫内膜腺上皮细胞与间质细胞胞浆着色基本一致 ;EM组增生期与分泌期子宫内膜均有乙酰肝素酶mRNA的表达 ,增生期阳性表达率为 83 3%( 10 / 12 ) ;分泌期为 72 7% ( 8/ 11) ,两者比较 ,差异也无显著性 (P >0 0 5 ) ;( 2 )对照组增生期子宫内膜乙酰肝素酶mRNA的阳性表达率为 4 1 7% ( 5 / 12 ) ,分泌期子宫内膜阳性表达率为 7 7% ( 1/ 13) ,两者比较 ,差异有显著性 (P <0 0 5 )。 ( 3)EM组子宫内膜乙酰肝素酶mRNA阳性表达率为 78 3% ( 18/2 3) ;对照组为 2 4 0 % ( 6 / 2 5 ) ,两者比较 ,差异也有显著性 (P <0 0 5 )。结论　乙酰肝素酶基因的表达与子宫内膜异位症的发病相关 ,其可能成为子宫内膜异位症治疗的一个有价值的靶点。     

紧密连接蛋白claudin-4在子宫内膜异位症患者在位与异位内膜组织中的表达

目的 检测紧密连接蛋白claudin-4在正常子宫内膜、子宫内膜异位症(内异症)患者的在位和异位内膜组织中的表达,评价elaudin-4在内异症发病中的作用.方法 选择35例内异症患者的卵巢异位内膜组织(异位内膜组),其中27例内异症患者的在位子宫内膜(在位内膜组)和35例非内异症良性卵巢、子宫疾病患者(对照组)的子宫内膜组织.透射电镜观察各组内膜组织中紧密连接的形态学变化;免疫组化和免疫印迹法检测elaudin-4蛋白的表达量,RT-PCR技术检测claudin-4 mRNA的表达量.结果 (1)透射电镜下,异位内膜组内膜组织中紧密连接结构消失,胶原组织丰富;在位内膜组内膜组织中紧密连接的结构与对照组内膜组织相似,未见明显变化.(2)免疫组化检测结果显示,clandin-4蛋白的阳性染色主要定位于子宫内膜腺上皮细胞膜,异位内膜组织中claudin-4的阳性染色很弱甚至缺失;在对照组、在位内膜组和异位内膜组内膜组织中,claudin-4蛋白表达量分别为89±24、84±22、27±14;claudin-4 mRNA表达量分别为14.5±6.8、13.8±9.5、2.6±2.5.异位内膜组内膜组织中claudin-4 mRNA和蛋白的表达量均低于在位内膜组和对照组内膜,差异有统计学意义(P<0.05).在位内膜组和对照组内膜组织中claudin-4 mRNA和蛋白的表达量分别比较,差异均无统计学意义(P>0.05).结论 异位子宫内膜组织中clandin-4的低表达可能参与了内异症的发病.     

单核细胞趋化蛋白-1在子宫内膜异位症患者子宫内膜组织中的表达

目的检测单核细胞趋化蛋白-1（MCP-1）在子宫内膜异位症(内异症)患者子宫内膜组织中的表达情况,并探讨其与内异症发病的关系。方法内异症患者30例作为研究组,非内异症患者20例作为对照组。用半定量逆转录聚合酶链反应（RT-PCR）方法检测两组子宫内膜组织中MCP-1信使核糖核酸(mRNA)的阳性表达率及相对水平。结果研究组26例(87%)内膜组织中可检测到MCP-1mRNA的表达,其中19例为强阳性表达,但其表达强度与内异症的严重程度无明显相关性。对照组15例(75%)内膜组织中可检测到MCP-1mRNA表达,仅3例为强阳性表达。研究组内膜组织中MCP-1mRNA的表达强度高于对照组(两组相对表达强度分别为0.746±0.345及0.460±0.341,P<0.05)。结论内异症患者的子宫内膜组织中MCP-1mRNA的表达增强,MCP-1对内异症的发病有一定促进作用。     

External validation of putative biomarkers in eutopic endometrium of women with endometriosis using NanoString technology

PurposeTo combine different independent endometrial markers to classify the presence of endometriosis.MethodsEndometrial biopsies were obtained from 109 women with endometriosis as well as 110 control women. Nine candidate biomarkers independent of cycle phase were selected from the literature and NanoString was performed. We compared differentially expressed genes between groups and generated generalized linear models to find a classifier for the disease.ResultsGeneralized linear models correctly detected 68% of women with endometriosis (combining deep infiltrating and ovarian endometriosis). However, we were not able to distinguish between individual types of endometriosis compared to controls. From the 9 tested genes, FOS, MMP7, and MMP11 seem to be important for disease classification, and FOS was the most over-expressed gene in endometriosis.Conclusion(s)Although generalized linear models may allow identification of endometriosis, we did not obtain perfect classification with the selected gene candidates.Electronic supplementary materialThe online version of this article (10.1007/s10815-020-01965-6) contains supplementary material, which is available to authorized users.     

子宫内膜异位症在位、异位内膜淋巴细胞亚群分布的研究

目的探讨子宫内膜异位症（内异症）在位和异位内膜淋巴细胞的分布及意义。方法采用免疫组织化学碱性磷酸酶抗碱性磷酸酶法,对43例内异症患者和19例子宫肌瘤等非内异症患者(对照组)的在位内膜和异位内膜中淋巴细胞及巨噬细胞亚群的数量、比例进行测定。结果内异症患者的增殖期异位内膜局部CD+3、CD+8、CD+68T淋巴细胞含量分别为（67.2±13.5）个/5HP（±s,下同）、（45.0±14.6）个/5HP、（37.2±10.6）个/5HP,明显多于在位内膜的（52.4±11.3）个/5HP（P<0.01）、（32.5±10.0）个/5HP（P<0.05）、（30.7±10.3）个/5HP,以及对照组内膜的（52.1±14.9）个/5HP（P<0.05）、（28.9±12.7）个/5HP（P<0.01）、（26.3±9.3）个/5HP（P<0.05）;分泌期CD+8T淋巴细胞/CD+4T淋巴细胞比值、CD+68T淋巴细胞含量分别为3.5±1.2、（40.3±12.2）个/5HP,高于对照组内膜的3.2±0.8（P<0.05）、（28.6±10.6）个/5HP（P<0.01）;巨噬细胞含量也明显增高。异位内膜中淋巴细胞无明显周期性改变。结论内异症病变局部淋巴细胞和巨噬细胞的大量侵润,与内异症的病理形成有关。在较晚期的内异症,淋巴细胞和巨噬细胞不能清除异位内膜,反之,可刺激其生长。     

Apoptosis patterns in eutopic and ectopic endometrium,adhesions and normal-looking peritoneum from women with or without endometriosis

Objective  To assess the apoptosis rate in eutopic and ectopic endometrial stromal and glandular cells, normal peritoneum and adhesions in women with endometriosis. Methods  A total number of 97 women with (n:60) and without (n:37) histopathologically confirmed endometriosis who underwent laparoscopy or laparotomy in the early follicular phase of the menstrual cycles for pain and infertility were included in this study. Stage I/II and stage III/IV were categorized as early staged and late-staged endometriosis. The endometrial samples were obtained with a Novack cannula from the corpus of the uterus. Normal-looking peritoneum, peritoneal implants and adhesions were sampled and fixed in formaldehyde for immunohistochemical staining with Bcl-2 and Bax. Tissue samples were fixed in formaldehyde for the assessment of apoptosis via terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and M30 cytoDEATH antibody. Results  The intensity of Bax staining of normal-looking peritoneum in early staged endometriosis was higher, compared to women with late-staged and women without endometriosis (P = 0.03). However, degree of Bcl-2 staining did not differ among early and late-staged endometriosis and women without endometriosis (P = 0.1). In terms of Bcl-2 and Bax staining in the stromal and glandular parts of the eutopic endometria, no significant differences were detected among three groups. In cases with early- and late-staged endometriosis the intensity of Bax and Bcl-2 stainings did not differ in both stromal and glandular parts of ectopic endometria. Number of cells with positive apoptotic signals assessed via TUNEL (P = 1.0) and M30 cytoDEATH antibody (P = 0.59) in normal-looking peritoneum did not differ between three groups. In addition, no difference in term of numbers of apoptotic cells obtained from adhesions was observed between three groups (for TUNEL, P = 0.29, for M30, P = 0.19). Conclusions  Apoptosis patterns did not differ in the eutopic and ectopic endometria as well as adhesions of women with or without endometriosis.     

血管内皮生长因子及血小板反应素在异位子宫内膜组织中的表达

目的：探讨血管内皮生长因子（VEGF）和血小板反应素（TSP）－1在子宫内膜异位症（内异症）患者异位及在位子宫内膜组织中的表达及其意义。方法：采用逆转录聚合酶链反应（RT－PCR）和Northern杂交法,检测38例内异症患者52份异位子宫内膜组织（腹膜红色病变12份,卵巢子宫内膜异位囊肿32份,宫骶韧带结节8份）,及30份在位子宫内膜组织（研究组）中VEGF mRNA及TSP－1 mRNA的阳性表达率,以25例非内异症患者的25份子宫内膜组织作为内异症患者在位内膜的对照（对照组）,结果：腹膜红色病变VEGF mRNA表达水平较高,TSP－1mRNA表达水平较低;卵巢子宫内膜异位囊肿VEGF mRNA表达水平较低,而TSP－1 mRNA表达水平较高,与对照组比较,研究组子宫内膜VEGF mRNA表达水平较高,而TSP－1mRNA表达水平较低。结论：内异症患者的异位子宫内膜中VEGF及TSP－1的表达,可能与其血管化程度有关,在位子宫内膜组织中VEGF及TSP－1表达水平的失衡,可能与内异症的生物学行为及发病有关。     

Relationship between apoptosis and the number of macrophages in eutopic endometrium from women with and without endometriosis

OBJECTIVE: To investigate the relationship between apoptotic cells and macrophages in the eutopic endometrium of women with and without endometriosis. DESIGN: Retrospective analysis of archival uterine endometrial biopsy specimens. SETTING: Institute for the Study and Treatment of Endometriosis, and university-based pathology and research laboratories. PATIENT(S): Fifty-one women with endometriosis and 24 healthy control subjects without endometriosis. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The number of TUNEL+ (terminal deoxynucleotide transferase [TdT]-mediated deoxyuridine triphospate [dUTP] nick end-labeling-positive) (apoptotic) cells and CD68+ (CD68 positive) (macrophages). RESULT(S): Apoptotic cells and macrophage numbers were positively correlated in the eutopic endometrium of women with and without endometriosis. However, the number of apoptotic cells and the macrophage content in the endometrium of women with endometriosis was significantly reduced compared with that of healthy control subjects without endometriosis. Differences between apoptosis and macrophage numbers between the two populations were observed predominantly during the early proliferative phase of the menstrual cycle. CONCLUSION(S): The reduction in apoptosis described for endometrial cells in women with endometriosis may be related to reduced macrophage trafficking into the eutopic endomtrium during the early-proliferative phase of the menstrual cycle.