连续服用低剂量复方炔诺酮与复方18-甲基炔诺酮对妇女脂质代谢的远期影响

目的 评价连续服用国产低剂量复方炔诺酮（复方炔诺酮）和低剂量复方１８－甲基炔诺酮（复方１８－甲）,对妇女脂质代谢的远期影响。方法 对连续服用复方炔诺酮（复方炔诺酮组,１６７例）和复方１８－甲（复方１８－甲组,１５０例）５～２５年及同期放置宫内节育器（ＩＵＤ）的健康妇女（对照组,１３１例）进行血清甘油三酯（ＴＧ）、总胆固醇（ＴＣ）、高密度脂蛋白（ＨＤＬ－Ｃ）、低密度脂蛋白（ＬＤＬ－Ｃ）、载脂蛋白Ａｌ（ＡｐｏＡｌ）、载脂蛋白Ｂ（ＡｐｏＢ）、脂蛋白ａ「Ｌｐ（ａ）」的测定。３组妇女的年龄、避孕药服用时间、体重指数、经济状况等,差异均无显著性。结果 与对照组比较,服用复方炔诺酮和复方１８－甲５～２５年妇女血清各项脂质水平均有不同程度的变化,两类口服避孕药使ＴＧ、ＨＤＬ－Ｃ显著升高;使ＡｐｏＡ１极显著升高。按不同避孕时间分     

应用分子杂交技术对“子宫颈癌高危患者”筛查及随访的临床——病…

目的：应用分子杂交技术对子宫颈癌高危患者进行筛查及随访的临床－病理学研究。方法：应用缺口翻译法＾３２Ｐ－标记的ｈＰＶ１６，１８，ｃ－Ｈａ－ｒａｓ－１及ｃ－ｍｙｃ－ＤＮＡ与子宫颈组织中的ＤＮＡ进行分子杂交，并通过Ｘ线胶片显示杂交结果，结果：慢性子宫颈炎组织中ｈＰＶ１６，１８，ｃ－Ｈａ－ｒａｓ－１及ｃ－ｍｙｃ阳性２９例，经１年随访，４例病情加重，２例手术治疗，２例保守治疗，４种高危因素均为阴性者随访２     

应用分子杂交技术对“子宫颈癌高危患者”筛查及随访的临床—病理学研究

目的：应用分子杂交技术对子宫颈癌高危患者进行筛查及随访的临床—病理学研究。方法：应用缺口翻译法３２Ｐ—标记的ｈＰＶ１６，１８，ｃ－Ｈａ－ｒａｓ－１及ｃ－ｍｙｃ－ＤＮＡ与子宫颈组织中的ＤＮＡ进行分子杂交，并通过Ｘ线胶片显示杂交结果。结果：慢性子宫颈炎组织中ｈＰＶ１６，１８，ｃ－Ｈａ－ｒａｓ－１及ｃ－ｍｙｃ阳性２９例，经１年随访，４例病情加重，２例手术治疗，２例保守治疗。４种高危因素均为阴性者随访２０例，均未发现病情加重。结论：用分子杂交技术筛查及随访子宫颈炎患者，对早期发现子宫颈癌具有重要意义。     

妊娠高血压综合征患者子宫螺旋动脉平滑肌细胞增生及c—myc？…

目的 探讨妊娠高血压综合征（妊高征）患者子宫螺旋动脉平滑肌细胞（ＶＳＭＣ）增生与原癌基因ｃ－ｍｙｃ表达的关系。方法 应用原位杂交方法对２０例血压正常妊娠妇女、１１例中度妊高征和１８你重度妊高征患者子宫螺旋动脉ＶＳＭＣ的ｃ－ｍｙｃ基因进行检测,同时观察子宫螺旋动态平滑肌层的组织结构变化。结果 中度和重度妊高征患者ＶＳＭＣｃ－ｍｙｃ的阳性表达率分别为６３．６％（７／１１）和７２．２％（１３／１８）,明     

应用聚合酶链反应技术检测子宫颈组织中人乳头瘤病毒的研究

应用聚合酶链反应（ＰＣＲ）技术对５３例正常宫颈组织、５９例宫颈炎、１２例宫颈癌、３例非典型增生及２例尖锐湿疣的子宫颈组织进行人乳头瘤病毒（ＨＰＶ）－１６，１８的检测。结果：正常子宫颈组织中ＨＰＶ－１６，１８的检出率低于宫颈癌组织，两者差异有显著意义；宫颈癌组织中ＨＰＶ－１６，１８的检出率高于宫颈炎组织，两者差异无显著意义。提示：部分有ＨＰＶ感染的宫颈炎可导致宫颈癌。     

子宫颈鳞癌组织中人乳头状瘤病毒E7蛋白与Rb基因产物的关系

目的：检测子宫颈鳞癌组织中是否存在人乳头状瘤病毒（ＨＰＶ）Ｅ７蛋白与Ｒｂ基因产物（ｐＲｂ）的结合物（ＨＰＶＥ７－ｐＲｂ）。方法：运用聚合酶链反应（ＰＣＲ）技术，对４０例宫颈鳞癌组织中ＨＰＶ６／１１、１６、１８、３３型ＤＮＡ进行检测，并用捕获酶联免疫吸附试验，探查ＨＰＶ感染的标本中有否ＨＰＶＥ７－ｐＲｂ存在。结果：４０例中，检出ＨＰＶＤＮＡ１８例（４５．０％）。１８例中９例存在ＨＰＶＥ７－ｐＲｂ，其中１例为ＨＰＶ１８，８例为ＨＰＶ１６。２例ＨＰＶ６／１１未检出ＨＰＶＥ７－ｐＲｂ。临床Ⅰ期患者ＨＰＶＥ７－ｐＲｂ的检出率高于Ⅱ～Ⅳ期患者（Ｐ＜０．０５）。ＨＰＶＥ７与ｐＲｂ的结合，与宫颈癌病理分级无关（Ｐ＞０．０５）。结论：在宫颈鳞癌组织中存在ＨＰＶＥ７－ｐＲｂ；ＨＰＶＥ７与ｐＲｂ的结合过程发生在宫颈癌变的较早阶段。     

放置释放左旋18—甲基炔诺酮宫内节育器前,后子宫内膜形态计量学…

本文选择３４例健康妇女，于放置释放２０μｇ／日左旋１８－甲基炔诺酮宫内节育器前与一年后子宫内膜作形态计量学研究，并与以前放置释放２μｇ／日左旋１８－甲基炔诺酮的宫内节育器子宫内膜作比较，结果表明：ＬＮＧ０ＩＵＤ－２０与ＬＮＧ－ＵＤ－２对子宫内膜增殖抑制和炎性反应有类似作用，其形态学变化无剂量依赖性影响，本研究结果结合临床资料说明：ＬＮＧ－ＩＵＤ－２０在不增加任何副作用的情况下起到了比ＬＮＧ－ＩＵＤ     

放置释放左旋18－甲基炔诺酮宫内节育器前、后子宫内膜形态计量学研究

本文选择３４例健康妇女，于放置释放２０μｇ／日左旋１８－甲基炔诺酮宫内节育器（ＬＮＧ－ＩＵＤ－２０）前与一年后子宫内膜作形态计量学研究，并与以前放置释放２μｇ／日左旋１８－甲基炔诺酮的宫内节育器（ＬＮＧ－ＩＵＤ－２）子宫内膜作比较。结果表明：ＬＮＧ－ＩＵＤ－２０与ＬＮＧ－ＩＵＤ－２对子宫内膜增殖抑制和炎性反应有类似作用，其形态学变化无剂量依赖性影响。本研究结果结合临床资料说明：ＬＮＧ－ＩＵＤ－２０在不增加任何副作用的情况下起到了比ＬＮＧ—ＩＵＤ—２更有效的作用。     

人绒毛膜促性腺激素和胰岛素对多囊卵巢综合征卵泡内 …

检验多囊卵巢综合重点中层得卵巢卵泡内膜细胞雄激素合成是否异常。方法对来自８例ＰＣＯＳ的卵巢卵泡与１８例正常妇女卵巢泡内膜细胞进行单层培养，采用放射免疫法测定两者在基础状态，人绒毛膜促性腺激素和胰岛素刺激作用下培养液中７１α－羟孕酮，雄烯二酮，孕酮。     

17α－乙炔基－17β－羟基－18－甲基－△~（4，15）－雌甾二烯－3－酮（Gestodene）的合成研究

以３－甲氧基－１７－羟基－１８－甲基雌甾－１，３，５（１０），８（９）－四烯（１１）为起始原料，经１１步反应制得Ｇｅｓｔｏｄｅｎｅ，总收率为３１％。运用澳化和脱溴化氢的化学方法引入关键的△１５双键结构，该方法所用试剂价廉易得，收率也较高，可作为Ｇｅｓｔｏｄｅｎｅ全合成的实用合成路线之一。     

应用套式聚合酶链反应技术检测孕妇血浆中胎儿DNA的研究

目的 依据孕妇血浆中存在游离胎儿ＤＮＡ的理论,寻找一种非创伤性产前基因诊断的新方法。方法 用套式聚合酶链反应技术对１２例１２￣４０孕周的初产妇血浆中游离胎儿ＤＮＡ进行特异性扩增,扩增的基因为Ｙ染色体短臂单拷贝基因片段（ＤＹＳ１４基因）,扩增片段的大小分别为２３９ｂｐ和１９８ｂｐ,１２例孕妇均采用母体血浆直接作为模板进行套式聚合酶链反应扩增。结果 １０例妊娠男性胎儿孕妇中有８例血浆中出现ＤＹＳ１４基     

Detection of Y chromosome-specific DNA in the plasma and urine of pregnant women using nested polymerase chain reaction

The present study was undertaken to evaluate a nested polymerase chain reaction (PCR) for detection of Y chromosome-specific fetal DNA in maternal plasma and urine of pregnant women during different gestational stages. DNA isolated from plasma and urine samples of 80 pregnant women (between 7 and 40 weeks' gestation) underwent amplification for Y chromosome-specific 198 bp DNA by nested PCR. The postpartum analysis of fetal gender showed that 55 women carried male and 25 female fetuses. Among the 55 women bearing male fetuses, Y chromosome-specific signals were detected in 53 (96%) plasma and 21 (38%) urine samples. Moreover, out of 25 women bearing female fetuses, 3 (12%) and 1 (4%) women had Y chromosome-specific signal in plasma and urine, respectively. Analysis of results with respect to gestational age revealed that there was no significant difference in the detection of Y chromosome-specific DNA between different trimesters in maternal plasma of women bearing male fetuses. These results showed that fetus-specific DNA was detected with high sensitivity (96%) and specificity (88%) in the maternal plasma by nested PCR, and therefore the method could be useful as a non-invasive procedure for fetal sex determination and prenatal diagnosis.     

Noninvasive genotyping of 9 Y-chromosome specific STR loci using circulatory fetal DNA in maternal plasma by multiplex PCR

BACKGROUND: Human Y-Chromosome specific STR (Y-STR) has now become a useful loci in casework. However, noninvasive genotyping of multiple Y-STR loci and its application in prenatal genetic diagnosis haven't been reported. The purpose of this study is to develop a Y-STR multiplex PCR amplification system that is suitable for the amplification of short-sized templates of circulatory male fetal DNA and use the established multiplex in noninvasive prenatal genetic diagnosis and its further applications in forensic casework. METHODS: On the basis of the characteristic of circulatory fetal DNA in maternal plasma, we selected 9 Y-STR loci in which the allele size was less than 180 bp in length and developed two multiplexes that allowed fluorescent genotyping of 9 Y-STR loci simultaneously. These Y-STR loci include two trinucleotide repeats (DYS426 and DYS388) and seven tetranucleotide repeats (DYS393, DYS460, H4; DYS391, DYS389 I, DYS456 and DYS458). Sixty-four pairs of plasma DNA samples from pregnant women and genomic DNA samples from their husbands were detected by our method. RESULTS: As a result, an average of 7.3 Y-STR specific alleles was detected in each of the 30 plasma DNA samples from pregnancies with male fetuses. However, none of these 9 Y-STR specific alleles was detected in 34 plasma samples from pregnant women carrying female babies. The chances of detecting Y-STR alleles ranged from 66.7 to 93.3%. Fifty-eight haplotypes were detected in 64 unrelated Chinese male individuals; haplotype diversity was 0.9966. This highly polymorphic Y-STR multiplex has greatly improved the chances of detecting the Y-STR allele. CONCLUSIONS: This assay provides a sensitive, accurate and efficient method for noninvasive prenatal genetic diagnosis and forensic casework.     

The best approach for early prediction of fetal gender by using free fetal DNA from maternal plasma

OBJECTIVES: Detection of free fetal DNA (ffDNA) in maternal blood during pregnancy has given rise to the possibility of developing new noninvasive approaches for early prenatal diagnosis.On a large-scale study, two protocols of real-time polymerase chain reaction (PCR) were compared in order to establish which Y-specific locus, either multicopy DYS14 or single copy SRY sequence, was the most suitable for developing a test with high diagnostic efficiency for early fetal gender assessment. The second aim was to assess whether the combination of the two detection systems could increase the performance of the prenatal test. METHODS: We analyzed 145 plasma samples from healthy pregnant women between 11 and 12 weeks of singleton gestation. For each sample, fetal gender was determined by using both protocols (DYS14 and SRY) during the same real-time PCR run. RESULTS: The data obtained by the DYS14 and SRY assays showed an efficiency in fetal gender prediction of 97.9 and 80%, respectively. It is not advisable to combine the two protocols because this association does not help in further improvements in fetal gender prediction. CONCLUSIONS: DYS14 assay is the best approach for early fetal gender assessment because it is more sensitive, accurate, and efficient than the SRY assay.     

Feasibility study of using fetal DNA in maternal plasma for non-invasive prenatal diagnosis

BACKGROUND: The discovery of the presence of fetal genetic material in maternal blood has opened up a new approach to prenatal diagnosis. One approach that has been extensively investigated over the past few decades is the isolation of fetal cells from maternal blood (Herzenberg et al. Proc Natl Acad Sci USA. 1979;76:1453-5; Bianchi et al. Proc Natl Acad Sci USA. 1990;87:3279-83; Cha et al. Prenat Diagn. 2005;25:586-91). As the fetal cells are scarce and the enrichment is of low efficiency, the technique could not be implemented in clinics. In 1997, Lo et al. (Lancet 1997;350(9076):485-7) discovered that cell-free fetal DNA is present in the plasma and serum of pregnant women. This discovery suggests that maternal plasma/serum DNA may be a useful source of material for non-invasive prenatal diagnosis. The objective of our study was to investigate the feasibility of using fetal DNA in maternal plasma for prenatal diagnosis. METHODS: Plasma DNA in 277 blood samples of 40 pregnant women at the gestational period from 5 to 40 weeks and 24 h after delivery were extracted by column separation. FQ-PCR was used to amplify the SRY sequence in 237 plasma samples of 30 pregnant women. Fluorescent PCR was used to amplify 9 short tandem repeat loci simultaneously in 40 plasma samples of 10 pregnant women, and genomic DNA samples from their husbands were amplified by the same method. RESULTS: The fetal SRY sequence could be detected from the 7th week of gestation, with a concentration that increased with progressing gestational age, attaining its highest peak before delivery. Twenty-four hours after delivery, fetal SRY sequence could not be detected in the maternal plasma. The concordance rate of the SRY sequence amplification results of plasma-free DNA, with real fetal gender was 100%. Analysis of maternal plasma samples collected during pregnancy revealed the presence of paternally inherited fetal-specific alleles. Among the 30 collected plasma samples, fetal-specific alleles were detected in 23 plasma DNA samples. The rate of positive results was 76% (23/30), and the frequency of positive results was 6/10 in early pregnancy, 8/10 in middle pregnancy, and 9/10 in late pregnancy. Short tandem repeats could not be detected from the maternal plasma 24 h after delivery. CONCLUSION: Fluorescent PCR can be used for amplification of fetal SRY sequence and STRs in maternal plasma to obtain fetal genetic information, which may have implications for non-invasive prenatal diagnosis of certain hereditary diseases.     

Earliest gestational age for fetal sexing in cell-free maternal plasma

OBJECTIVES: To evaluate at what gestational age fetal DNA can reliably be detected at the earliest in maternal plasma. METHODS: We performed consecutive blood sampling in the first trimester of pregnancy in 17 women who were pregnant after in vitro fertilization (IVF) or intrauterine insemination (IUI). DNA was isolated and the Y-chromosome specific SRY was amplified by real-time polymerase chain reaction (PCR).We likewise studied 31 women prior to invasive prenatal diagnosis procedures for test validation purposes. All test results were compared to cytogenetic sex or sex at birth. RESULTS: The earliest SRY detection was at a gestational age of 5 weeks and 2 days. In none of 4 pregnancies ending in a miscarriage was SRY detected. We detected SRY in maternal plasma in 1 of 2 patients (50%) carrying a male fetus at a gestational age of 5 weeks, in 4 of 5 (80%) at a gestational age of 7 weeks, in 4 of 4 (100%) at a gestational age of 9 weeks. In all 7 women pregnant with a male fetus, the correct fetal sex was detected by 10 weeks. In none of the 6 patients who delivered a girl was SRY detected. In the validation group, SRY was detected in 13 of the 13 male, and none of the 18 female fetuses. CONCLUSIONS: We conclude that real-time PCR of the SRY gene promises to be a reliable technique for early fetal sexing in maternal plasma.     

经母血采集胎儿细胞行产前诊断的最佳时间探讨

目的：探讨利用母血循环中胎儿细胞进行产前诊断的最佳采血时间。方法：对４１例孕龄为６～１４周的妇女连续取血，采用套式聚合酶链反应技术检测人类Ｙ染色体特异的锌指蛋白基因（ＺＦＹ）。结果：１９例妊娠男性胎儿妇女外周血ＺＦＹ随着孕龄的增加，其胎儿单拷贝基因的检出率增高，其中孕６周时检出率为１／１９（５．３％），孕１１周时为１３／１９（６８．４％），而到孕１４周时，则达到１８／１９（９５．０％）；对２２例妊娠女性胎儿妇女外周血进行ＺＦＹ检测时，无一例假阳性结果，这一检测方法在妊娠早期进行胎儿性别鉴定的总准确率达到９７．８％（４０／４１）。结论：利用母血循环中胎儿细胞进行产前诊断的最佳采血时间应在妊娠１４周，同时提示胎儿细胞最早进入母血循环中的时间在不同个体间存在明显的差异。     

Influence of gestational age on fetal deoxyribonucleic acid retrieval in maternal peripheral blood

OBJECTIVE: We wanted to verify whether gestational age influences the retrieval of fetal deoxyribonucleic acid in maternal blood to identify the best period for maternal blood sampling for a future noninvasive prenatal diagnoses. STUDY DESIGN: We amplified 81 deoxyribonucleic acid samples extracted from the peripheral blood of 27 pregnant women (18 bearing male fetuses and 9 bearing females) by nested polymerase chain reaction of the Y-specific sequence DYS14. We obtained three blood samples (one per gestational trimester) from each woman. Statistical evaluation was assessed by the McNemar test of symmetry. RESULTS: Polymerase chain reaction results in male-bearing pregnancies differed significantly between the first and second trimesters and between the second and third trimesters (p < 0.025) in parallel with a decrease in sensitivity in the second trimester (67%) compared with the first (94%) and third trimesters (100%). CONCLUSIONS: The drop in sensitivity from the first to the second trimester witnesses a variable concentration of fetal cells in maternal blood, with a negative balance in the second trimester. Therefore, to achieve an adequate polymerase chain reaction accuracy, the choice of gestational age is relevant and the first trimester seems to be more suitable than the second trimester.(Am J Obstet Gynecol 1997;177:22)     

Direct quantification of fetal cells in maternal blood by real-time PCR

BACKGROUND: Fetal cells in maternal blood still present an enticing alternative for the development of a safe and efficacious non-invasive method for prenatal diagnosis. However, most enrichment methods are very tedious and have failed to realise this long sought after goal. We developed a simple, robust TaqMan real-time PCR assay to directly quantify male fetal cells in maternal blood using the multi-copy DYS14, without the need for any additional enrichment procedure. METHODS: The sensitivity of the DYS14 assay was evaluated by using female genomic DNA spiked with male DNA or male cells. The specificity of the DYS14 assay was evaluated by examining 40 adult blood samples and 44 maternal blood samples from pregnant women with known fetal gender. Direct quantification of fetal cells in maternal blood was performed in 14 of the maternal blood samples by the DYS14 assay. The results were compared to those by SRY assay. RESULTS: The sensitivity of DYS14 PCR assay was found to be higher than that of SRY assay for the detection of fetal cells. The number of fetal cells in maternal blood did not exceed 2/mL blood. The presence of cell-free fetal DNA in maternal blood could lead to erroneous quantification of fetal cell DNA. The number of fetal cells detected in blood cell pellets, which had been unwashed to remove the cell-free fetal DNA, was indeed significantly higher than those in extensively washed samples. CONCLUSIONS: We quantified the fetal cells in maternal blood without fetal cell enrichment procedures by TaqMan real-time PCR for a multi-copy DYS14 locus. Our assay is sensitive and also suitable for automation, and may be a useful tool for the determination of fetal-maternal cell trafficking, such as microchimerism.     

Prenatal fetal sex diagnosis by detecting amelogenin gene in maternal plasma

OBJECTIVES: To provide a new, reliable noninvasive method for fetal sex determination. METHODS: Fetal sex was detected in 32 early pregnant women by identifying the amelogenin gene in maternal plasma using nested PCR analysis. First, the 122/128 bp of X-Y homologous region containing 6 bp deletions in the intron 3 of amelogenin gene in X chromosome was amplified, and then the nested PCR was carried out, whose 3' end of the upstream primer is just located in the deletion region. The fetus was male or female, depending on whether it had the 89-bp nested PCR product or not. RESULTS: The 89 bp of nested PCR product was detected in 19 plasma samples obtained from pregnant women, deducing they bear the male fetus and the remaining pregnant women bear female. When compared with the birth outcome, two samples were pseudo-positive. The coincidence was 93.8%. This method had high sensitivity that even trace amount of target fetal DNA (10 pg) could be detected. CONCLUSIONS: This conventional nested PCR analysis of amelogenin gene promises to be a reliable method for noninvasive fetal sex determination at early pregnancy using maternal plasma DNA.