复发性流产绒毛整合素β1的表达及染色体分析

目的探讨复发性流产与绒毛染色体异常及整合素β1的表达的关系。方法将在我院就诊的45例早孕期复发性流产患者作为研究组,30例正常早孕行人工流产术的患者作为对照组,对两组患者进行绒毛细胞染色体分析及绒毛组织整合素β1的检测。结果研究组绒毛染色体异常发生率为46.66%,显著高于对照组（10.00%）;研究组绒毛组织整合素β1的表达率为22.22%,显著低于对照组（86.67%）;研究组：染色体正常者整合素β1的表达率低于染色体异常者,但两者经统计学处理无显著差异（χ^22.81 P（0.05）;在对照组：染色体正常者整合素β1的表达率与染色体异常者整合素β1的表达率经统计学处理无显著差异（确切概率P=0.360）;两组间染色体正常者整合素β1的表达率有显著差异,研究组整合素β1表达率显著低于对照组（χ^229.75 P（0.05）;两组间染色体异常者整合素β1的表达率经统计学处理无显著差异（确切概率P=0.533）。结论绒毛染色体异常是导致孕早期复发性流产的主要原因,绒毛组织整合素β1表达缺失与绒毛染色体检查正常的流产有关。     

Two siblings with unusually mild homozygous β-thalassemia: A didactic example of the effect of a nonallelic modifier gene on the expressivity of a monogenic disorder

Two adult Black sibs with homozygous β-thalassemia had severe deficiency of β-chain production but an unusually mild clinical course and almost normal hematocrit values. Their father had the typical hematological findings of β-thalassemia trait but the mother, in addition to the β-thalassemia trait, had elevated F cells (42.2%). Elevated F cells were also present in a hematologically normal sister of the affected providing evidence that a gene for heterocellular hereditary persistence of fetal hemoglobin (HPFH) was also running in this family. The unusually mild clinical course of the two homozygotes is attributed to their inheritance from their mother of the heterocellular HPFH gene. The gene for heterocellular HPFH is nonallelic to the β-globin locus and apparently acted as a modifier of the homozygous β-thalassemia phenotype by facilitating F-cell production and thus diminishing the pathophysiological consequences of the β-thalassemia defect. The effect of heterocellular HPFH on the expression of homozygous β-thalassemia or Hb S genotypes is an impressive example of the pathophysiological basis of changes in the expressivity of monogenic disorders due to action of nonallelic modifiers.     

Pineal Serotonin Metabolism in Non-Innervated Perinatal Glands before and after Intraocular Maturation: Supersensitivity of Adrenoceptors that have never been Innervated

Transplantations were made of fetal pineal glands (crown-rump length, CRL, 19–30 mm) or pineal glands from adult male rats to the anterior chamber of the eye of the rat. Studies were performed with regard to the importance of the age of the donor animal (and thereby the degree of maturation and innervation of the gland to be transplanted) for the possible development of denervation supersensitivity. The transplants were cultured in a medium containing ¹⁴C-serotonin. Increased production of ¹⁴C-N-acetylserotonin (NAcS) was used as the main criterion for β-adrenergic stimulation. 4 experimental groups were obtained by transplanting fetal or adult pineals to intact or sympathetically denervated eyes. In all 4 groups β₁-stimulation (KWD 2033 10^-6 M) increased ¹⁴C-NAcS formation. The response to β-stimulation was significantly higher in denervated fetal pineal transplants than in innervated fetal transplants and thus demonstrating β-receptor supersensitivity. It was concluded that a) the ability to respond to β-adrenoceptor stimulation with increased ¹⁴C-NAcS formation develops between the 18th and 20th day of gestation, b) transplants derived from fetal as well as from adult rats can respond to β-adrenergic stimulation, c) this sensitivity also develops in oculo in transplants that at the time of transplantation lacked the capacity to increase their ¹⁴C-NAcS formation in response to treatment with β-agonist, d) denervation supersensitivity occurs in fetal transplants that became mature in sympathetically denervated eyes.     

Anti-phospholipid induced murine fetal loss: novel protective effect of a peptide targeting the β2 glycoprotein I phospholipid-binding site. Implications for human fetal loss

β2 glycoprotein I (β2GPI)-dependent anti-phospholipid antibodies (aPL) induce thrombosis and affect pregnancy. The CMV-derived synthetic peptide TIFI mimics the PL-binding site of β2GPI and inhibits β2GPI cell-binding in vitro and aPL-mediated thrombosis in vivo. Here we investigated the effect of TIFI on aPL-induced fetal loss in mice. TIFI inhibitory effect on in vitro aPL binding to human trophoblasts was evaluated by indirect immunofluorescence and ELISA. TIFI effect on aPL-induced fetal loss was investigated in pregnant C57BL/6 mice treated with aPL or normal IgG (NHS). Placenta/fetus weight and histology and RNA expression were analyzed. TIFI, but not the control peptide VITT, displayed a dose-dependent inhibition of aPL binding to trophoblasts in vitro. Injection of low doses of aPL at day 0 of pregnancy caused growth retardation and increased fetal loss rate, both significantly reduced by TIFI but not VITT. Consistent with observations in humans, histological analysis showed no evidence of inflammation in this model, as confirmed by the absence of an inflammatory signature in gene expression analysis, which in turn revealed a TIFI-dependent modulation of molecules involved in differentiation and development processes. These findings support the non-inflammatory pathogenic role of aPL and suggest innovative therapeutic approaches to aPL-dependent fetal loss.     

1026例孕中期妇女唐氏综合征和神经管缺陷的血清筛查及细胞染色体分析

目的：探讨孕中期妇女（14～20）周外周血甲胎蛋白（AFP）和游离绒毛膜促性腺激素（f-βHCG）在胎儿唐氏综合征（Down’s syndrome,DS）产前筛查中的作用。方法：应用免疫化学发光法分别检测1026例孕中期孕妇血清AFP和f-βHCG浓度,结合母龄、孕周和体重等因素,用筛查软件计算风险系数,对DS高危孕妇进行羊水细胞染色体核型分析,神经管缺陷（NTD）高危孕妇行B超检查。结果：筛查的1026例孕妇中,DS高危孕妇64例,阳性率6.2%,NTD高危孕妇9例,阳性率0.88%。在高危孕妇中发现4例胎儿染色体异常,1例经B超检查确诊为神经管畸形。两组异常胎儿染色体检出率有显著性差异（P〈0.01）,35岁或35岁以上孕妇组胎儿异常发生率高于35岁以下孕妇组（P〈0.01）。结论：利用DS筛查结合B超产前诊断,能够提高胎儿先天性缺陷的早期发现和诊断。     

Associations between maternal and fetal serum levels of immune activation markers and fetal growth

During pregnancy, signs of maternal immunologic sensitization to fetal HLA and other fetoplacental alloantigens are often detectable in peripheral blood. Presumbly, this in part reflects immune activity at the maternal-fetal interface. This response may involve activation of maternal T cells, which stimulate placental growth via lymphokine production. To shed light on this mechanism, data on placental weight, neonatal anthropometry, gestational age, fetomaternal HLA relationships (reflecting a potential for HLA allosensitization), and serum levels of three immune activation markers in maternal and cord blood were collected in a sample of 61 primiparous women and their neonates. The activation markers were soluble CD8 antigen (sCD8), interleukin-2 receptor (sIL-2R), and beta-2 microglobulin (β₂m). Mean fetal and maternal sCD8 and β₂m levels, and mean fetal sIL-2R levels were significantly higher than published norms. Fetal means for all three markers exceeded maternal means, and both sIL-2R and β₂m were highly correlated between mother and fetus. This suggests that fetal sIL-2R and β₂m levels result in part from transport or diffusion from the maternal compartment. No associations were found between fetomaternal HLA relationships, activation markers, and placental weight. The difference between the fetal and maternal β₂m value was significantly correlated with birth weight, controlling for chest circumference and crown-heel length. Associations between birth weight and fetomaternal HLA relationships could not be interpreted with certainty. These findings suggest that maternal immune activation and diffusion or transport of β₂m into fetal compartment enhances fetal growth in fat-free body mass. © 1995 Wiley-Liss, Inc.     

Senescence is accelerated through donor cell specificity in cloned pigs

Animals cloned by somatic cell nuclear transfer (SCNT) sometimes have abnormalities that result in large offspring syndrome or early death during gestation due to respiratory and metabolic defects. We cloned pigs using two sources of donor cells and observed phenotypic anomalies in three pigs cloned from one type of cell, s-pig fetal fibroblasts. These animals had many wrinkles on their faces and bodies and looked older than age-matched normal pigs. We performed the present study to examine whether the wrinkled phenotype in the cloned pigs was due to senescence, a genetic problem with donor specificity, or epigenetic problems with reprogramming. To address this issue, we investigated biomarkers of senescence, including telomere length and the expression of senescence-associated β-galactosidase (SA-β-gal), glyceraldehyde phosphate dehydrogenase (GAPDH) and β-actin. We also assessed the methylation status of euchromatic PRE-1 repetitive sequences and centromeric satellite DNA, and measured the mRNA levels of six imprinted genes, Copg2, Mest, Igf2R, GNAS, SNRPN and Ube3a. The telomeres of the wrinkled cloned pigs were much shorter than those of the normal cloned pigs and age-matched normal pigs. In the wrinkled cloned pigs, SA-β-gal activity was detected and GAPDH and β-actin were repressed. The mRNA levels of Mest, GNAS and Ube3a were reduced in the wrinkled cloned pigs, although there was no difference between the normal cloned pigs and normal controls. This gene expression analysis indicates that the wrinkled abnormality of our pigs originates from genetic abnormalities in the donor cells used for SCNT.     

Effect of Antiphospholipid Antibodies on β2Glycoprotein I-Phospholipid Interaction

PROBLEM: β₂ glycoprotein I (β₂GPI) physiologically binds to negatively charged phospholipids (PLs) and is a natural regulator of the coagulation cascade. Thrombotic clinical complications and recurrent fetal loss associated with autoimmune antiphospholipid (aPL) antibodies are thought to be related to their binding to β₂GPI-PL complex and interference with the physiological function of β₂GPI. METHOD OF STUDY: To investigate the effect of aPL on β₂GPI-PL interaction, we studied the binding of biotinylated β₂GPI to cardiolipin (CL) by enzyme-linked immunosorbent assay (ELISA) in the presence and absence of purified aPL immunoglobulin G (IgG) antibodies. RESULTS: Adding five different aPL IgG antibodies with different levels of aPL activity isolated from the sera of five patients with aPL-associated recurrent fetal death greatly increased the binding of biotinylated β₂GPI to CL-coated plates. The optical densities (ODs) were 0.635, 0.890, and 1.265 in the presence of three aPL IgG antibodies, compared to 0.425 in the absence of aPL IgG. In contrast, normal human IgG had no enhancing effect. The OD was 0.480 and 0.425, respectively. The enhancement of β₂GPI binding to CL by aPL IgG correlated with the titers of aPL antibodies. The use of phosphate-buffered saline with increasing salt concentrations as a washing buffer for the ELISA resulted in more stable binding of β₂GPI to PL in the presence of aPL IgG. CONCLUSIONS: These findings suggest that the binding of autoimmune aPL antibodies to β₂GPI-PL complex results in abnormally tighter interaction between β₂GPI and PLs, which may lead to physiological dysfunction of β₂GPI as a regulator of coagulation.     

小鼠胎肝间质干细胞体外向肌样细胞分化的研究

目的：了解小鼠胎肝间质干细胞在体外向肌样细胞分化的潜能。 方法： 无菌条件下从正常C57BL/6J胎鼠肝脏中分离出间质干细胞，体外培养传3代后用5-氮胞苷和二性霉素B诱导分化，观察胎肝间质干细胞诱导前后形态变化；用RT-PCR检测细胞诱导前后, 成肌调控基因Myf5和成肌素myogenin的表达情况；诱导后7 d和 14 d 行免疫细胞化学染色鉴定诱导后细胞结蛋白和α-肌动蛋白的表达。 结果： RT-PCR显示诱导后6 h至72 h成肌调控基因Myf5和myogenin序贯表达，而诱导前的细胞则没有检测到表达；免疫细胞化学染色提示第7 d细胞结蛋白和α-肌动蛋白表达阳性，第14 d阳性率明显升高。 结论： 胎肝中分离出的间质干细胞在体外可以定向诱导分化为肌样细胞。     

Inhibition by anti-beta2-microglobulin of MLR by human fetal liver cells

Liver cells of fetuses without lymphoid thymus show strong, immunologically nonspecific proliferation when confronted with adult foreign lymphocytes. To elucidate the mechanism of this response, we have studied the effect of anti-β₂-microglobulin on fetal liver cells in mixed lymphocyte reaction (MLR). Anti-β₂-microglobulin inhibited MLR when antiserum was present in the culture or when the responding fetal cells had been preincubated with it. If the adult lymphocytes stimulating fetal liver cells were preincubated with anti-β₂-microglobulin, only a slight or no effect was observed. No difference was observed between liver cells from fetuses with and without lymphoid thymus. β₂-microglobulin was also demonstrated on the fetal liver cells by immunofluorescence. The results reveal that β₂-microglobulin is present on early fetal liver cells, and that MLR by fetal liver cells is inhibited by anti-β₂-microglobulin in the same way as in adults. multipotential immunocompetence of early fetal liver cells cannot be excluded on the basis of these experiments.     

Atlas of Wnt and R-spondin gene expression in the developing male mouse lower urogenital tract

Prostate development is influenced by β-catenin signaling, but it is unclear which β-catenin activators are involved, where they are synthesized, and whether their mRNA abundance is influenced by androgens. We identified WNT/β-catenin-responsive β-galactosidase activity in the lower urogenital tract (LUT) of transgenic reporter mice, but β-galactosidase activity differed among the four mouse strains we examined. We used in situ hybridization to compare patterns of Wnts, r-spondins (Rspos, co-activators of β-catenin signaling), β-catenin-responsive mRNAs, and an androgen receptor-responsive mRNA in wild type fetal male, fetal female, and neonatal male LUT. Most Wnt and Rspo mRNAs were present in LUT during prostate development. Sexually dimorphic expression patterns were observed for WNT/β-catenin-responsive genes, and for Wnt2b, Wnt4, Wnt7a, Wnt9b, Wnt10b, Wnt11, Wnt16, and Rspo3 mRNAs. These results reveal sexual differences in WNT/β-catenin signaling in fetal LUT, supporting the idea that this pathway may be directly or indirectly responsive to androgens during prostate ductal development.     

脐血毛细管电泳在产前诊断重型地中海贫血中的应用

目的探讨脐血毛细管电泳在产前诊断重型地中海贫血中的应用价值。方法对71例夫妻双方患有同型地中海贫血的孕妇进行脐静脉穿刺术,抽取脐血同时进行地中海贫血基因检测和血红蛋白毛细管电泳。结果重型α地中海贫血胎儿脐血Hb Bart’s含量高达72.9%∽89.8%,而Hb H病、轻型α地中海贫血及静止型α地中海贫血胎儿脐血Hb Bart’s含量分别为18.4%∽24.3%、2.1%∽5.7%及0.3%∽0.6%,正常胎儿脐血中未检出Hb Bart’s。重型β地中海贫血胎儿脐血Hb A含量极低,为0%∽0.6%,而轻型β地中海贫血胎儿和正常胎儿脐血Hb A含量分别为2.2%∽5.2%和2.5%∽9.4%。脐血Hb Bart’s和Hb A定量分析能准确的检出胎儿重型地中海贫血。结论毛细管电泳具有操作简便及自动化的优点,能准确诊断胎儿重型地中海贫血,可以作为产前诊断的一种常规方法。     

利用孕妇外周血浆中小片段游离胎儿DNA进行β-地中海贫血无创性产前诊断

目的利用cffDNA对胎儿进行广西、广东地区常见的17种β-地中海贫血基因型的检测,与传统创伤性产前诊断相比较,探讨该方法的准确性和可行性。方法针对广西、广东地区常见的β-地中海贫血突变的基因型设计三对不同引物,并用生物素标记。对cffDNA进行二次PCR反应后,使用反向斑点杂交(revert dot-blot hybridization,RDB)检测胎儿的β-珠蛋白基因型,与创伤性产前诊断结果比较,观察准确率。结果37例cffDNA标本检测中,检出重型β-地中海贫血19例,轻型β-地中海贫血11例,正常7例,与创伤性产前诊断结果相比较出现3例误诊,准确率为91.9%(34/37)。结论利用cffDNA进行β-地中海贫血的检测,可能出现母源性DNA背景的污染,当胎儿基因型与母亲基因型相同时,必须提高警惕,进一步分析或者复查。因为该技术取样容易,对孕妇胎儿无风险,不受孕期时间影响,易为与孕妇接受,因此进一步改进该技术后有望可用于β-地中海贫血的诊断。     

一体化灌注法体外构建活化人工骨的优化研究

目的探讨人胚成骨细胞在多孔β-磷酸三钙（β-TCP）支架内灌注性接种及培养的影响因素及其作用机制,对一体化灌注法体外构建活化人工骨进行优化研究。方法应用自行设计的灌注式生物反应器进行多孔β-TCP支架内人胚成骨细胞的一体化灌注性接种和培养,以静态接种为对照。通过细胞活力（Mrrr法）测定、活细胞接种率、组织形态学观察和计量学分析等分别检测细胞接种时间、接种密度、灌注速率等因素对支架内细胞黏附和生长的作用。结果细胞灌注接种效果优于静态接种;灌注接种-灌注培养法细胞生长及分布优于静态接种-灌注培养法。接种时间、接种密度、灌注速率等因素均显著影响细胞接种及培养效果。灌注法构建活化人工骨的最优条件：接种时间12h～24h,接种密度2×10^5／ml一5×10^5／ml,灌注接种速率1ml／min,灌注培养速率0．5ml／min～2ml／min（灌注初期24h）及2ml／min（灌注24h后）。结论一体化灌注法利于多孔β-TCP支架内人胚成骨细胞的黏附和生长。接种时间、接种密度、灌注速率等因素均影响活化人工骨体外构建效果,对其进行优化有助于人工骨移植物的临床转化和推广应用。     

Acute blockade of β1-receptors in the asphyxiated sheep fetus

Acute blockade of β₁-receptors in the asphyxiated sheep fetus. Acta Physiol Scand 130 , 381–385. Received 5 November 1986, accepted 9 February 1987. ISSN 0001–6772. Department of Paediatrics, Landspitalinn, University Hospital, Reykjavik, Iceland and Department of Physiology and Department of Paediatrics I, University of Goteborg, Sweden. The effects of acute β₁-blockade on fetal cardiovascular reactions during asphyxia were evaluated in 11 exteriorized sheep fetuses. Gestational age was 110–142 days. Asphyxia was induced either by ventilating the mother with low oxygen gas mixture or by mechanical reduction of placental blood flow. During asphyxia all fetuses reacted to metoprolol injection with a decrease in heart rate, myocardial contractility, cardiac output and arterial blood pressure. Five experiments resulted in irreversible fetal cardiovascular collapse. Isoprenaline was given to the fetuses during hypoxia to test the ability to further increase heart rate and activate myocardial β-adrenoceptors. In those experiments with fetal cardiovascular demise after metoprolol, the isoprenaline injection did not result in a significant tachycardia. The surviving fetuses could increase their heart rate as a sign of a capacity to further increase the sympatho-adrenergic drive.     

大鼠不同发育阶段β-防御素-2基因

目的:探讨大鼠β-防御素-2基因在肺组织表达的发育调控。方法:根据Genbank大鼠β-防御素-2的cDNA序列,设计一对特异引物。采用RT-PCR技术在大鼠不同发育阶段的肺组织总RNA中,扩增其特异cDNA片段,并进行DNA序列测定。借助Genbank中BLAST程序,将从该cDNA序列推导的氨基酸序列与人的hBD-2和大鼠的rBD-1做相似性分析。结果:从足月胎鼠、出生8h和4d以及成年大鼠相同重量肺组织所提取的总RNA中,均扩增出一条密度相同、长度约为220bp的cDNA片段。在所推导氨基酸序列中与hBD-2氨基酸相同率为48.6%(17/35),与rBD-1氨基酸残基的相同率为31%(11/35)。其成熟分子链长35个氨基酸残基,含β-防御素共有的且排列次序相同的6个典型的半胱氨酸和其它保守氨基酸残基。结论:本实验表明不同发育阶段大鼠肺组织均表达β-防御素-2mRNA,提示该分子可能是肺组织天然抵抗微生物感染的一个重要分子基础。     

福州地区正常孕妇妊娠期标准化尿HCG、β-HCG变化曲线

绘制福州地区正常孕妇妊娠期＂标准化＂尿HCG、β-HCG变化曲线图。对180名正常孕妇妊娠540周的尿液,用R IA法检测尿HCG、β-HCG,用酶法检测尿肌酐并计算各孕周经尿肌酐校正后的标准化尿HCG、β-HCG值,并绘制HCG、β-HCG变化曲线。结果表明福州地区正常孕妇妊娠期标准化尿HCG、β-HCG变化曲线的拟合相关系数分别为0.628和0.702,拟合有意义（P〈0.05）。本文正常孕妇妊娠期标准化尿HCG、β-HCG整体变化曲线、血清和尿HCG的变化都与文献报道基本一致。     

Mucopolysaccharidosis VII as cause of fetal hydrops in early pregnancy.

We report on fetal hydrops presenting at 18 weeks of gestation and diagnosed as beta-glucuronidase deficiency. The parents were first cousins and there were 2 previous similar fetal deaths. beta-Glucuronidase was absent in cultured fetal fibroblasts and lymphoblasts but was normal in the tested relatives. The activities of other lysosomal enzymes were normal.