米非司酮对中期引产母、儿外周血SCE的影响

目的探讨用不同剂量米非司酮（MFT）中期妊娠引产对母、儿姐妹染色单体交换（SCE）频率的影响,评估MFT对人体的染色体稳定性影响。方法选择符合研究条件的中期引产患者120例,随机分为4组：空白对照组,分别口服MFT 150、200、300 mg引产组,于胎儿娩出后取母亲静脉血及胎儿脐带血,进行SCE频率检测比较。结果①母亲组MFT 150 mg组、200 mg组与对照组SCE频率比较差异无统计学意义;300 mg组与对照组SCE频率比较,差异有统计学意义（P〈0.001）;各组均随着用药剂量加大,SCE频率逐渐升高（spearmanr=0.802,P〈0.05.）。②胎儿各组脐血SCE频率与是否用药以及药物剂量差异均无统计学意义（P〉0.05）。结论①小剂量MFT（150、200 mg）引产,不引起母亲SCE频率显著增加。但300 mg引产时,SCE频率显著提高,应引起临床注意。②不同剂量MFT对胎儿SCE频率无影响。     

服用“复方炔雌醚”妇女的外周血淋巴细胞姐妹染色单体互换的研究

本文研究长效口服避孕药复方炔雌醚(内含炔雌醚3.3毫克和氯地孕酮15毫克)对妇女外周血淋巴细胞姐妹染色单体互换(SCE)率的影响。结果,服药妇女每个细胞SCE率平均值为3.55±0.41(S.E),和正常妇女的3.22±0.41(S.E)比较,两者差异没有统计学意义(P>0.05)。表明复方炔雌醚不具有明显的DNA 初级损伤效应。     

染色体不稳定性及HPV、CMV感染与宫颈癌发病的关系

目的 :探讨染色体数目畸变、脆性位点发生率、姐妹染色单体交换发生率、细胞内微核发生率、宫颈分泌物内人乳头状瘤病毒 (HPV)感染、巨细胞病毒 (CMV)感染与子宫颈癌的关系。方法 :对 30例子宫颈癌患者 ,做外周血细胞培养 ,分析染色体畸变、姐妹染色单体交换、脆点和微核。用PCR方法扩增宫颈分泌物HPV16 -DNA、HPV18-DNA、CMV -DNA。结果 :染色体畸变 ,姐妹染色单体交换发生率 ,脆性位点、细胞内微核发生率与正常对照组差异有显著性 ,宫颈癌组HPV16 -DNA、HPV18-DNA和CMV -DNA感染阳性率分别为 50 %、46 .67%和 53 .33 % ,3者均为阳性者占 2 0 %。结论 :染色体不稳定性增加及HPV、CMV感染与宫颈癌发病相关     

子痫前期患者血管内皮祖细胞数量及生物学功能的变化

目的:观察子痫前期患者血管内皮祖细胞(endothelial progenitor cells,EPCs)数量与功能的变化。方法:选取孕28~40周子痫前期孕妇、健康孕妇各20例,抽取外周静脉血20m l,经密度梯度离心法分离单个核细胞,通过CD133/CD34双荧光标记及Ⅷ因子鉴定细胞;用流式细胞仪检测细胞数量,用MTT比色法、改良Boyden小室法、黏附能力测定实验,分别观察EPCs的增殖、迁移及黏附功能的变化。结果:与健康孕妇相比,子痫前期患者外周血EPCs数量明显减少(4.29%±1.21%vs15.32%±2.00%,P<0.01),其增殖能力(增殖率13.45%±1.68%vs 18.45%±1.67%)、迁移能力[迁移细胞(37.25±7.28)个/视野vs(67.10±9.55)个/视野]、黏附能力[黏附细胞(20.65±5.19)个/视野vs(34.40±6.72)个/视野]明显受损。结论:子痫前期患者外周血EPCs数量减少、生物学功能减退。     

非孕妇和孕妇子宫肌与外周血淋巴细胞β_2－受体相关性研究

目的观测非孕妇和正常孕妇子宫肌与外周血淋巴细胞β２－肾上腺素能受体（β２－ＡＲ）结合量的变化及两者的相关性。方法以［３Ｈ］－ＤＨＡ（３Ｈ－双氢心得舒）为放射配基，分别测定１０例行子宫切除术的非孕妇及１０例剖宫产者子宫肌和外周血淋巴细胞β２－受体结合量。结果１．非孕妇子宫肌与外周血淋巴细胞β２－ＡＲ结合量呈良好相关性（ｒ＝０．７５６８，Ｐ＜０．０１）。２．孕妇子宫肌与外周血淋巴细胞β２－ＡＲ结合量亦呈良好相关性（ｒ＝０．８５２９，Ｐ＜０．０１）。３．孕妇子宫肌与淋巴细胞β２－ＡＲ结合量均较非孕妇组明显下降。结论淋巴细胞β２－ＡＲ结合量可以反映并监测子宫肌β２－ＡＲ结合量的变化，作为细胞生物学指标指导围产期医学管理和临床治疗。     

Cytokine production in natural killer cells and lymphocytes in pregnant women compared with women in the follicular phase of the ovarian cycle

OBJECTIVE: To test whether peripheral natural killer (NK) cells, helper T cells, and cytotoxic lymphocytes of pregnant women shift from a type 1 cytokine production toward a type 2 cytokine production as compared with these cells in women in the follicular phase. DESIGN: Prospective study. SETTING: Outpatient clinic. PATIENT(S): Healthy nullipara at 30 weeks' amenorrhea and healthy nonpregnant women in their follicular phase. INTERVENTION(S): Samples of whole blood were stimulated with phorbol myristate acetate (PMA; Sigma Chemical Co., St. Louis, MO) and Ca-ionophore in the presence of monensin (Sigma). Lymphocytes were stained with alpha-CD3, alpha-CD8, and alpha-interferon gamma (IFN-gamma) alpha-interleukin 2 (IL-2), IL-4, or IL-10. Analysis was performed by flow cytometry. Statistical evaluation was done with the Mann-Whitney U test. MAIN OUTCOME MEASURE(S): Percentage NK cells, helper lymphocytes, and cytotoxic lymphocytes that were producing IFN-gamma, IL-2, IL-4, or IL-10. RESULT(S): There is a statistically significant decrease in the percentage of NK cells, and helper and cytotoxic lymphocytes that produced IFN-gamma in pregnant women when compared with women in the follicular phase. There is also a statistically significant decrease in the percentage of helper lymphocytes producing IL-2 in pregnant women compared with nonpregnant women. CONCLUSION(S): We found a decrease in type 1 cytokine production with no change in type 2 cytokine production after in vitro stimulation of "pregnant" NK cells and lymphocytes as compared with "nonpregnant" NK cells and lymphocytes. We suggest that NK cell and lymphocyte response are shifted away from a type 1 immune response during pregnancy.     

正常妊娠外周血组织多肽抗原与完整hCG关系的探讨

目的 :测定正常妊娠妇女外周血中组织多肽抗原 (TPA)和完整hCG的浓度 ,探讨两者变化的关系。方法 :使用TPA酶免试剂盒与双抗体夹心法定量测定血清中TPA的含量。应用时间分辨荧光免疫分析法 (time resolvedfluorescenceimmunoassay)测定hCG的浓度。结果 :正常妊娠期外周血TPA的浓度随妊娠进展逐步增加 ,晚孕期最高。与早、中孕期及非孕期比较差异均有显著性 (P <0 .0 5 )。血清完整hCG浓度早孕期明显高于其它各期 (P <0 .0 5 )。结论 :外周血TPA水平随妊娠进展逐步增加可能代表滋养细胞凋亡的情况 ;hCG浓度与滋养细胞增殖相关。TPA、hCG对预测某些病理妊娠可能有帮助。     

The influence of paternal lymphocytes immunization on percentage of peripheral blood CD16+/CD56+ cells in women with primary recurrent spontaneous abortion

OBJECTIVES: Paternal lymphocytes immunization has been proposed for several years as an efficient treatment for unexplained Recurrent Spontaneous Abortion (RSA), however precise mechanism that underline the benefits of this immunotherapy is still unclear. DESIGN: The aim was to study the influence of paternal lymphocytes immunization on percentage of peripheral blood NK cells (CD16+/CD56+) in women with primary RSA of unknown etiology. MATERIALS AND METHODS: 48 patients with history of 3-5 (mean 3.3 +/- 0.7) consecutive primary RSA of unknown etiology were selected for the study. Immunotherapy with paternal lymphocytes, isolated from 100 ml of peripheral blood, was performed twice prior conception with a 4-week interval. The percentage of NK cells (CD16+/CD56+) was estimated using standard flow-cytometric immunofluorescent techniques for whole blood with one-step monoclonal anti-CD16/CD56 antibodies. Statistical analysis was performed with Wilcoxon test and the p value less than 0.05 was accepted as statistically significant. RESULTS: It was found that paternal lymphocytes immunization significantly decreases the percentage of NK cells in peripheral blood in women with RSA (23.9 +/- 8.5 vs. 16.2 +/- 7.0; p = 0.009). CONCLUSIONS: The data of the present studies suggest that paternal lymphocytes immunization modulate immunity in women with primary unexplained RSA significantly decreasing the percentage of NK cells (CD16+/CD56+) in peripheral blood.     

Tumor-infiltrating lymphocytes contain higher numbers of type 1 cytokine expressors and DR+ T cells compared with lymphocytes from tumor draining lymph nodes and peripheral blood in patients with cancer of the uterine cervix

OBJECTIVE: The aim of this study was to compare the phenotype and function of lymphocytes collected from the peripheral blood (PBL), tumor draining regional lymph nodes (LND), and infiltrating tumor tissues (TIL) of patients with stage IB-IIA cervical cancer. METHODS: Leukocytes from peripheral blood (n = 35), tumor draining lymph nodes (n = 33), and tumor tissues (n = 15) of cervical cancer patients were evaluated for the relative proportions of lymphocyte subsets including CD3+, CD4+, CD8+, CD19+, CD56, and the early (CD25) and late (HLA-DR) activation markers on CD3+ T cells, as well as the ability to synthesize type 1 cytokines (IFN-gamma and IL-2) and a type 2 cytokine (IL-4) by flow cytometry. RESULTS: In all patients, T cells (CD3+) were the major leukocyte population detected in each tissue, with CD4+ T cells being dominant in PBL and LND, while CD8+ T cells predominated in TIL (CD4:CD8 ratios, 2.4 vs 4.0 vs 0.7, respectively). CD19+ lymphocytes (B cells) were significantly higher in LND compared to PBL and TIL (P > 0.01) while CD56+ lymphocytes were higher in PBL compared to LND (P > 0.01) and TIL (P > 0.05). The early activation marker CD25 was significantly up-regulated in LND, while TIL had a higher proportion of T cells expressing the late activation marker HLA-DR. Type 1 cytokines were the dominant type produced by in vitro stimulated T cells for each population, with a greater proportion of IFN-gamma+ CD4+ and CD8+ T cells (i.e., Th1 and Tc1) and IL-2+ CD8+ T cells (Tc1) seen in TIL, as compared with LND and PBL (P > 0.01). Low percentages of IL-4+ T cells (i.e., Th2 and Tc2) were detected only in PBL. CONCLUSIONS: This study demonstrates significant differences in the phenotype and activation state of lymphocyte subsets from different anatomical sites, as well as differences in their ability to synthesize immunostimulatory cytokines. The recruitment and accumulation of high concentrations of antigen-experienced T lymphocytes in the cervical tumor tissue may represent an important local barrier to neoplastic dissemination.     

卵巢癌组织及外周血淋巴细胞多药耐药基因表达蛋白的定量研究

目的探讨卵巢癌组织及化疗期间腹水癌细胞（ＡＣＣ）与外周血淋巴细胞（ＰＢＬ）多药耐药基因（ＭＤＲ１）表达蛋白Ｐ１７０含量及其相关性。方法应用流式细胞免疫学方法,对４８例卵巢癌组织细胞、ＡＣＣ及ＰＢＬ的Ｐ１７０含量进行动态定量研究。结果术前２５％卵巢癌组织Ｐ１７０含量阳性;化疗期间ＰＢＬ及ＡＣＣＰ１７０含量均逐渐增高。结论卵巢癌细胞不仅可以原发表达Ｐ１７０,且可随化疗疗程的增加而耐药逐渐增强,ＰＢＬ的Ｐ１７０含量可间接反映残余或复发卵巢癌细胞对化疗药物耐受的程度     

原因不明复发性流产患者调节性T淋巴细胞对树突状细胞的调控作用

目的 探讨原因不明复发性流产(URSA)患者外周血和蜕膜组织中CD+4CD+25调节性T淋巴细胞(Tr细胞)对树突状细胞(DC)的调控作用.方法 采集4例URSA患者(流产组)和4例正常早孕妇女(对照组)的外周血和蜕膜组织,密度梯度离心法、免疫磁珠分离法分选出Tr细胞和DC,培养6 d,培养方法分为DC单独培养和Tr细胞与Dc混合培养.酶联免疫吸附试验(ELISA)检测两种方法培养的细胞培养上清液中辅助性T淋巴细胞(Th)1型细胞因子--γ干扰素(IFN-γ)的Th2型细胞因子--白细胞介素10(IL-10)的蛋白含量.结果 (1)外周血:流产组细胞混合培养和单独培养后,细胞培养上清液中IFN-γ蛋白含量为(22.5±3.0)、(23.2±0.7)ng/L,两者比较,差异无统计学意义(P>0.05);IL-10蛋白含量分别为(35±4)、(37±7)ng/L,两者比较,差异也无统计学意义(P>0.05).对照组细胞混合培养和单独培养后,IFN-γ蛋白含量为(36±1.1)、(30.5±4.0)ns/L,两者比较,差异无统计学意义(P>0.05);IL-10蛋白含量分别为(36±9)、(54±20)ng/L,两者比较,差异有统计学意义(P<0.01).(2)蜕膜组织:流产组细胞混合培养和单独培养后,IFN-γ蛋白含量分别为(24.4±2.5)、(23.4±2.6)ng/L,两者比较,差异无统计学意义(P>0.05);IL-10蛋白含量分别为(25±5)、(28±7)ng/L,两者比较,差异也无统计学意义(P>0.05);对照组细胞混合培养后,IFN-γ蛋白含量为(22.6±3.8)ng/L,明显低于单独培养后的(30.7±4.6)ng/L,差异有统计学意义(P<0.01);对照组细胞混合培养后,IL-10蛋白含量为(31±9)ng/L,明显高于单独培养后的(27±6)ng/L,差异也有统计学意义(P<0.05).结论 URSA患者Tr细胞抑制性免疫功能下降,从而导致对DC调控失常,Th1/Th2平衡失调和母胎免疫耐受异常.     

孕妇血清、乳汁及新生儿脐血中输血传播病毒感染的研究

目的 了解输血传播病毒（TTV）在孕妇血清、乳汁及新生儿脐血中的感染状况,探讨TTV的传播途径。方法 用半巢式聚合酶链反应方法对150例孕妇血清、乳汁及新生儿脐血标本,进行TTV DNA扩增,对8例PCR阳性扩增产物再进行直接测序。结果 （1）150例孕妇血清TTV DNA的阳性率为11．3％（17／150）,17例孕妇血清TTV DNA阳性者中,乳汁阳性11例,其新生儿脐血均为阴性。（2）检测3例孕妇血清TTV DNA序列及乳汁TTV DNA序列,其核苷酸同源性为97．6％－99．2％;5例孕妇血清TTV DNA序列之间同源性为71．5％－92．8％,与日本原型株NSS的核苷酸及氨基酸序列同源性为71．5％－92．8％。结论 孕妇血清及乳汁中存在TTV感染,经乳汁传播可能是该病毒感染的主要方式之一。