抗人上皮性卵巢癌单克隆抗体的制备与鉴定

应用卵巢浆液性乳头状囊腺癌可溶性相关抗原ＣＡ＿（９２５）免疫ＢＡＬＢ／ｃ小鼠，与ＮＳ－１骨髓瘤细胞株融合制各单克隆抗作（ＭｃＡｂ），获得５株持续性分泌抗体的细胞株，以融合孔为代号，分别为ＯＣ＿（４Ｄ９），ＯＣ＿（７Ｅ１０），ＯＣ＿（１Ｂ４），ＯＣ＿（３Ｂ８），ＯＣ＿（９Ｂ９）。５种ＭｃＡｂ的滴度分别为１：１０￣７，１：１０￣７，１：１０￣６，１：１０￣４，１：１０￣５，免疫球蛋白亚型为ＩｇＭ和ＩｇＧ型，染色体呈双亲染色体特点。所获得５种ＭｃＡｂ与ＣＡ＿（９２５）有特异性反应（阳性反应）；与正常卵巢组织及子宫平滑肌瘤未呈阳性反应；与肺癌、胃癌、肾癌、食管癌、直肠癌组织有弱阳性反应。     

重组白细胞介素－1β对小鼠胚胎着床的影响

本文报道了重组白细胞介素－１β（ｒＩＬ－１β）对小鼠胚胎着床的影响，并对其作用机理进行初步的探讨。研究证实，ｒＩＬ－１β能够有效地阻断妊娠第３、４天小鼠胚胎着床的发生，此外，ｒＩＬ－１β能致小鼠早期胚胎发育异常，运行滞缓，并改变小鼠胚泡植入阶段受卵巢激素调控的子宫内膜有丝分裂模式，还诱发着床前子宫腔白细胞浸润现象。上述结果表明，ｒＩＬ－１β对抗着床效应是多因素的协同作用。     

重组白细胞介素—1β对小鼠胚胎着床的影响

本文报道了重组白细胞介素－１β（ｒＩＬ－１β）对小鼠胚胎着床的影响，并对其作用机理进行初步的探讨。研究证实，ｒＩＬ－１β能够有效地阻断妊娠第３、４天小鼠胚胎着床的发生，此外，ｒＩＬ－１β能致小鼠早期胚胎发育异常，运行滞缓，并改变小鼠胚泡植入阶段受卵巢激素调控的子宫内膜有丝分裂模式，还诱发着床前子宫腔白细胞浸润现象。上述结果表明，ｒＩＬ－１β对抗着床效应是多因素的协同作用。     

癌基因bcl—2及抑癌基因p53在子宫内膜癌的表达及临床 …

目的：研究癌基因ｂｃｌ－２及抑癌基因ｐ５３在子宫内膜癌发生，发展中的作用，方法：采用免疫组化ＡＢＣ法检测４９例子宫内膜癌中ｂｃｌ－２ｐ５３基因蛋白的表达，结果：４９例子宫内膜癌中的２６例ｂｃｌ－２表达阳性，占５３％１２例ｐ５３表达阳性，占２５％，子宫内膜组织学分级Ｇ１，Ｇ２，ｂｃｌ－２表达率（６６％）显著高于Ｇ３表达率（２１％，Ｐ〈０．０５），而Ｇ３ｐ５３表达率（４６％）显著高于Ｇ１，Ｇ２表达率（     

国内近期有关文献题录

基础理论０４７５４　阳　豫　李东至等：细胞凋亡与胎盘的发育和成熟。生殖医学杂志８（３）：１９０，１９９９０４７５５　李红真　郑淑蓉等：整合素与生殖。生殖医学杂志８（３）：１８６，１９９９０４７５６　朱　瑾　周剑萍等：小鼠胚泡着床前后子宫内膜细胞间粘附分子－１ｍＲＮＡ表达水平的变化。上海医科大学学报２６（５）：３６５，１９９９０４７５７　陆金春　黄宇烽等：精子受精抗原（ＦＡ－１）的研究进展。男科学报５（３）：１６６，１９９９０４７５８　胡冬梅　陈竹钦等：外源性细胞因子对早孕蜕膜调节自然杀伤细胞活性…     

子宫内膜癌bcl-2癌基因的持续性表达及其临床意义

目的：研究子宫内膜癌ｂｃｌ－２癌基因的表达及其临床意义。方法：采用免疫组化ＡＢＣ法检测增生期、分泌期、单纯型增生、复合型增生及不典型增生子宫内膜共２６份，子宫内膜癌４９例的ｂｃｌ－２癌基因蛋白表达及雌、孕激素受体（ＥＲ、ＰＲ）的表达。结果：正常增生期子宫内膜、增生的子宫内膜存在ｂｃｌ－２的表达，与ＥＲ相关，分泌期子宫内膜ｂｃｌ－２表达下降；４９例子宫内膜癌中２６例ｂｃｌ－２表达阳性，占５３％，２９例ＥＲ表达阳性，占５９％，２５例ＰＲ表达阳性，占５１％。７２％ｂｃｌ－２表达阳性者ＥＲ阳性，７５％ｂｃｌ－２表达阴性者ＥＲ阴性（Ｐ＜０．０１）。６８％ｂｃｌ－２表达阳性者ＰＲ阳性，６２％ｂｃｌ－２阴性者ＰＲ阴性（Ｐ＜０．０５）。子宫内膜癌Ｇ１、Ｇ２级ｂｃｌ－２的表达率为６６％，显著高于Ｇ３级者（２１％）（Ｐ＜０．０５）。ｂｃｌ－２的表达与肌层浸润、手术分期无关，ｂｃｌ－２表达阳性及阴性者生存率统计差异无显著性。结论：子宫内膜ｂｃｌ－２的持续性表达与卵巢激素相互作用可能在子宫内膜癌发生、发展中起作用     

中草药促排卵汤对小鼠卵巢及子宫内膜作用的组织化学研究

昆明系雌性小鼠在生育期服用相当于人剂量的中草药促排卵汤５ｄ后，取出其卵巢和子宫，制成冰冻切片，均用油红０法显示脂类（Ｌ），用Ｌｏｊｄａ法显示３－β－羟基甾体脱氨酶（３－β－ＨＳＤＨ），用亮氨酰－β萘酰胺法显示氨基肽酶（ＬＮＡｓｅ），用偶氮偶联法显示碱性磷酸酶（ＡＫＰ），对子宫还用ＰＡＳ法显示糖原（ＧＬｙ）。结果表明，与对照组相比，无论是动情前期或动情后期，服用中草药的小鼠卵巢其卵泡内膜层及颗粒层细胞和黄体细胞的胞浆内３－β－ＨＳＤＨ活性和脂类明显增高；子宫内膜上皮细胞及间质内的糖蛋白和脂类明显增高。对结果的讨论说明中草药促排卵汤确有促进卵巢卵泡和黄体的甾体激素分泌，并可增强子宫内膜营养的作用，这些都会有利于胚泡的着床及发育。     

着床期兔胚泡对子宫内膜糖蛋白影响的研究

本文应用ＳＤＳ－ＰＡＧＥ，蛋白质免疫印渍技术及以凝集素为探针对一侧输卵管结扎兔着床期子宫内膜糖蛋白进行了研究。结果表明１．ｃｏｎＡ－ｂｉｎｄｉｎｇＧＰｓｏｓ非着床侧略高于着床侧。２．ＷＧＡ－ｂｉｎｄｉｎｇＧＰ着床侧各条带扫描后峰值均高于非着床侧。３．分子量为９７．３ＫｄＰＮＡ－ｂｉｎｄｉｎｇＧＰｗ和８５．９ＫｄＰＮＡ－ｂｉｎｄｉｎｇＧＰＳＤＳ着床侧明显高于非着床侧，而３８．５ＫｄＰＮＡ－ｂｉｎｄｉｎｇＧＰＳＤＳ非着床侧则明显高于着床侧，４．分子量为９７．３Ｋｄ和ｇｏ．６ＫｄＲＣＡ－ｂｉｎｄｉｎｇＧＰＳＤＳ着床例明显高于非着床侧，而３７．２ＫｄＲＣＡ－ｂｉｎｄｉｎｇＧＰＳＤＳ非着床例明显高于着床侧．显示在同一激素状态下，胚泡着床与否影响到子宫内膜糖蛋白出现了不同变化。     

白细胞介素对小鼠体外受精及早期胚胎发育的影响

本文就人白细胞介素（ＩＬ－１β、５、６）对小鼠的体外受精率、体外受精卵的发育率，以及从交配雌鼠体内回收的２细胞期胚（体内受精卵）的发育率带来的影响进行了探讨。ＩＬ－１β５０ｎｇ／ｍｌ、ＩＬ－５５ｎｇ－ｍｌ以上、ＩＬ－６０．５ｎｇ／ｍｌ以上时对小鼠的体外受精率有明显抑制（Ｐ＜０．０５）。体外受精卵序的发育仅ＩＬ－６实验组见到抑制，０．５ｎｇ／ｍｌ以上对４细胞期胚发育有抑制，５０ｎｇ／ｍｌ时对８细胞期胚到囊胚期的发育有抑制。对体内受精卵的发育ＩＬ－１β和ＩＬ－６实验组与对照组间均未见到差异，但ＩＬ－５对囊胚期的发育有促进。以上结果提示，子宫内膜异位症时，人腹腔液中存在的ＩＬ－１β、５、６由于其抑制受精率而造成不孕。     

子宫内膜整合素蛋白β3亚基在妊娠中的变化

为探讨整全素蛋白β３亚基在胚胎着床期的作用，本实验应用特异性抗整合素蛋白ヂ３抗体，采用免疫和Ｗｅｓｔｅｒｎ即迹法，二氢基联苯胺显色，ＶＤＳ扫描等技术，检测妊娠ｄ３、５、７、１５小鼠子宫内膜及人分泌期子宫内膜和妊娠早期中整合素β３亚基的表达量。结果：小鼠子宫内膜整合素蛋白Ψ３亚基表达量在妊娠ｄ５明显上升，至ｄ廛达高峰（与对照组比较，Ｐ〈０．０１）。此后下降，ｄ１５时，整合素蛋白β３的表达量与对照级相     

肿瘤转移抑制因子CD9/MRP-1基因在小鼠围着床期子宫内膜的动态表达

目的:探讨肿瘤转移抑制基因(CD9)/运动相关蛋白(MRP-1)mRNA和蛋白在胚胎着床过程中的作用。方法:应用RT-PCR和免疫组化技术观察CD9/MRP-1mRNA和蛋白在早孕和假孕小鼠子宫中的表达规律。结果:早孕d1-4小鼠子宫组织均有CD9/MRP-1mRNA表达,且d4表达最多;其蛋白主要表达在d1-4的子宫内膜上皮细胞,且早孕d2-4CD9/MRP-1在子宫基质细胞散在阳性表达。假孕d1-8小鼠子宫组织均有CD9/MRP-1mRNA表达,假孕d5表达开始增加,至d6达到峰值。而CD9/MRP-1蛋白在假孕d1-8子宫内膜腺上皮均有表达,而子宫内膜腔上皮均无表达。假孕d2-5,子宫基质细胞出现散在阳性表达。结论:①CD9/MRP-1在早孕小鼠子宫中呈动态表达,提示它在胚胎精确侵袭子宫内膜的调节中发挥作用。②CD9/MRP-1在妊娠小鼠子宫的表达是非胚胎依赖性的。     

肿瘤坏死因子α转换酶(TACE)在早孕小鼠子宫内膜的表达规律

目的:探讨肿瘤坏死因子α转换酶(TACE)在胚胎植入过程中的作用。方法:①用RT-PCR及免疫组化方法分别检测未孕(n=10)及孕d1-7小鼠(n=10×7)子宫内膜TACE mRNA和蛋白的表达;②向妊娠小鼠(n=6)左侧子宫角注射TACE抗体,观察TACE对胚胎着床数的影响。并以右侧为自身对照和注射生理盐水为手术对照组(n=6)。结果:①TACE mRNA在早孕期小鼠子宫内膜的表达明显高于未孕期,且随妊娠天数的增加而逐渐增加,于孕d4达高峰,随后渐降。免疫组化结果与RT-PCR一致;②子宫角TACE抗体注射后胚胎植入数目明显减少。结论:在小鼠妊娠早期,TACE表达增高,其在胚泡植入中可能发挥着重要作用。     

PCNA在早孕小鼠子宫内膜的表达

目的:探讨增殖细胞核抗原(PCNA)mRNA及蛋白在早孕小鼠子宫内膜的作用。方法:实时荧光定量PCR(FQ-PCR)及免疫组化方法分别检测未孕(d0)及早孕d2、d3、d4、d5、d6、d7小鼠子宫内膜PCNA mRNA和蛋白的表达。结果:随着小鼠妊娠天数的增加,PCNAmRNA及蛋白表达量也逐渐增高,在孕d4、d5达到峰值,孕d6、d7逐渐下降。结论:在胚泡植入窗口期子宫内膜PCNA的高表达,可能参与了子宫内膜蜕膜化过程,在胚泡早期植入中发挥作用。     

肿瘤转移抑制因子CD82/KAI1基因在小鼠子宫内膜的动态表达

目的：探讨肿瘤转移抑制因子CD82/KAI1在小鼠胚胎着床过程中的功能。方法：应用RT-PCR和免疫组织化学技术分别观察CD82/KAI1mRNA和蛋白在小鼠动情周期和早孕子宫中的表达。结果：在动情周期中,CD82/KAI1mRNA除在动情期较低外,其余三期的表达量基本相同,但蛋白质却是在动情期表达最丰富,子宫内膜上皮和基质均呈阳性。在早孕子宫中,CD82/ KAI1mRNA的表达逐渐增多,蛋白质表达的量和范围也逐渐增强。结论：CD82/KAI1在小鼠子宫中呈动态表达,提示它在胚胎精确侵袭子宫内膜的调节中发挥作用。     

TRAIL在小鼠胚胎着床过程中子宫内膜的表达

目的:检测TRAIL在小鼠胚胎着床过程中子宫内膜的表达,探讨它在蜕膜细胞凋亡中的作 用。方法:采用RT-PCR及免疫组化技术检测妊娠d 1-8小鼠子宫组织TRAILmRNA及蛋白的表 达情况。结果:妊娠d 1-8的小鼠子宫组织均有TRAIL mRNA的表达,且着床期间的表达较着床前 明显增加(P<0．05)。妊娠d 1-3,小鼠子宫内膜无TRAIL蛋白表达;妊娠d 4,TRAIL表达在小鼠胚 胎定位、黏附点的子宫内膜腔上皮细胞;妊娠d 5-6,TRAIL定位于胚胎着床点附近的蜕膜细胞中; 妊娠d 7-8,TRAIL表达在与子宫蜕膜邻近的胚胎滋养层细胞中。结论:在小鼠胚胎着床过程中, TRAIL诱导子宫内膜腔上皮细胞凋亡可能是胚胎跨越上皮屏障的重要机制之一,且TRAIL诱导的 蜕膜细胞和胚胎滋养层细胞的凋亡在滋养层细胞对子宫内膜的适度侵入过程中起重要作用。     

Expression of antigens involved in the presentation of lipid antigens and induction of clonal anergy in the female reproductive tract

The molecular backgrounds of the anti-phospholipid syndrome and immunisation against the Rhesus proteolipid antigens are still largely unknown. In the present study, expression of (1) CD1, a major histocompatibility complex class l-like lipid antigen presenting molecule, (2) IL-10, a cytokine promoting induction of clonal anergy, and (3) CD80 and CD86, two T-cell costimulators preventing induction of clonal anergy when present, was investigated in frozen sections of cervix, corpus and the fallopian tube (FT) of 25-day-old BALB/c mice injected with FSH, progesterone or medium and of pregnant mice from each trimester (days 7, 14 and 19). CD1 was expressed by all endometrial epithelial cells. Enhanced immunostaining of the endometrial epithelial cells was observed after FSH treatment, and cervix and FT expressed generally more than the corpus of the uterus. After treatment with medroxy progesterone acetate (MPA), expression of CD1 by the endometrial epithelia was weak. During pregnancy CD1 was absent from the endometrium adjacent to the foetus, but was unaltered in the cervix and FT. IL-10 was expressed by the endometrial glands and also by the endometrial surface epithelium. MPA treatment increased the intensity of the IL-10 immunofluorescence. There were also chains of positive cells between the muscle bundles within the pregnant myometrium. CD80 and CD86 were usually absent from the female reproductive tract, but were occasionally found in the cervix during pregnancy. The present study demonstrates definite differences in the expression of both CD1 and IL-10 between the FSH and MPA treated mice, suggesting differences during the oestrous cycle. As IL-10 is expressed more in the secretory phase, it is probably involved in making the endometrium more acceptable for implantation by inducing clonal anergy. This is supported by the absence of CD80 and CD86. These results also suggest that abnormal expression of CD1d during pregnancy may predispose the mother for immunisation against lipid antigens such as membrane phospholipids and Rhesus-antigens.     

Distribution of some Gal beta 1-3(4)GlcNAc related carbohydrate antigens on the mouse uterine epithelium in relation to the peri-implantational period

Using monoclonal antibodies of defined carbohydrate specificity we have looked at the distribution of various Gal beta 1-3(4)GlcNAc related oligosaccharide determinants in the mouse uterus during the first 6 days of pregnancy. Frozen sections of uterus from B6D2F1, B6CBF1 or B6D2F1/BOM female mice were incubated with the monoclonal antibodies and then with a fluorescein isothiocyanate conjugate of goat anti-mouse IgM and viewed by epifluorescence illumination. None of the antibodies bound specifically to stroma cells but antibodies recognising difucosylated Gal beta 1-3(4)GlcNAc structures, the monofucosylated type II determinant (SSEA-1) and an H type I oligosaccharide bound to cells of the uterine luminal epithelium and glands and to the uterine secretions. Antibodies recognising the three different types of saccharide showed independent changes in staining intensity during early pregnancy. The antibody which recognises H type I structures (667/9E9) showed a change in distribution from binding to most cells of the uterine epithelium in the non-pregnant mouse and on day 3 of pregnancy to binding restricted to areas of epithelial cells interspersed with non-staining clumps of cells between days 4 and 5 of pregnancy.     

Ia antigen expression in the rat uterus

Using indirect immunofluorescence and monoclonal antibodies, we have determined the distribution of Ia+ cells in the non-pregnant rat uterus. In cycling animals, Ia+ cells, most of dendritic shape and non-specific esterase negative, were abundant in the connective tissue of the myometrium and throughout the endometrium, especially adjacent to the luminal and glandular epithelia. In contrast, the number of T lymphocytes was very much less and few of these cells expressed Ia antigen. Stromal Ia+ cells were more numerous during estrus than in any other phase of the cycle. Only a few Ia+ cells were seen in neonatal uterine samples, but the numbers increased to approximately adult levels by 32 days of age. Prepubertal glands were consistently negative, but epithelial expression of Ia antigens in adult uteri showed interesting variations. The luminal epithelium was always negative, whereas the glandular epithelial cells often were positive. Glandular staining was quite heterogeneous in individual samples, but most gland profiles were negative or weakly positive during proestrus and estrus and positive during diestrus. Ovariectomized females given different steroid supplements revealed significant hormonal effects. Estrogen alone dramatically increased the numbers of Ia+ stromal cells compared with control samples, but progesterone alone had no effect. However, progesterone given with estrogen abrogated the estrogenic influence. Progesterone alone caused decreased expression of Ia antigens by the glandular epithelium. If the highly-Ia/ cells in the stroma are capable of antigen presentation, these results indicate a significant potential for local antigen processing with the uterine endometrium.     

sLe~x在小鼠胚泡植入早期子宫内膜中的表达及作用

目的:探讨唾液酸化的路易斯寡糖(sLex)在胚泡植入早期小鼠子宫内膜的表达及作用。方法:①应用免疫组化方法对sLex在小鼠妊娠早期子宫内膜的表达进行定位,免疫印迹法以另一侧为自身对照,检测妊娠d1-5小鼠子宫内膜sLeX的表达情况。②向妊娠小鼠一侧子宫角注射sLeX单克隆抗体,观察其对胚胎着床的影响。结果:妊娠d1-5小鼠子宫内膜组织中,sLeX表达量逐渐增加,在植入窗口期(d4)达高峰;子宫角注射抗体后对胚泡植入有明显的抑制作用。结论:在小鼠胚泡植入早期,sLex参与了胚泡的植入过程,在胚泡和子宫内膜的识别过程中发挥重要的作用。     

HLA antigen on the trophoblast of normal pregnancy

The mechanism of fetal survival as a semiallograft in the uterus remains to be clarified. In this context, the expression of HLA antigen on the trophoblast which stands between the mother and the fetus is the main problem, because HLA antigen plays an important role in immunological reaction to an allograft. For this purpose, 41 pregnant uteri (6-18 weeks of gestation) were examined immunohistochemically (avidin-biotin-peroxidase complex method) using monoclonal antibodies to HLA antigens. Troma 1, a rat monoclonal antibody, was used as a trophoblast marker in immunohistochemical studies. The results were as follows: HLA-A,B,C is not expressed on syncytiotrophoblast and villous cytotrophoblast. HLA-A,B,C is expressed on nonvillous cytotrophoblast which exists in cytotrophoblastic cell column, cytotrophoblastic shell, and endometrium. HLA-DR is not expressed on any trophoblast. The absence of HLA antigen on syncytiotrophoblast and villous cytotrophoblast seems to be essential for the survival of the fetus. But it is proved that some trophoblasts express HLA-A,B,C on their cell surface and they are adjacent to endometrial cells or maternal blood. From these findings, it seems that fetal HLA antigen might be recognized by the mother and there might be an exquisite immune escape mechanism in decidua.