应用巢式PCR扩增孕妇血浆中胎儿SRY基因的研究

目的:建立一种巢式聚合酶链反应(PCR),利用孕妇血浆中的游离胎儿DNA(fetal DNA)鉴定胎儿SRY基因。方法:随机采集30例孕妇外周血标本,采用酚/氯仿法从孕妇血浆中提取DNA,设计两对引物对SRY基因进行巢式PCR扩增,扩增产物经测序加以确认。结果:17例孕男胎的孕妇血浆中有15例经巢式PCR扩增检出SRY基因,而13例孕女胎的孕妇血浆没有检出阳性结果。准确性和敏感性分别为93.3%(28/30)和88.2%(15/17)。结论:应用酚/氯仿法从孕妇血浆中提取游离DNA简单有效,结合巢式PCR扩增SRY基因技术可用于无创性产前性连锁遗传性疾病的诊断。     

血游离DNA检测及肿瘤的基因诊断（文献综述）

肿瘤病人血中有异常升高的游离肿瘤DNA,用分子生物学方法检测病人血游离DNA中肿瘤相关基因的异常表达,是无创伤性诊断肿瘤的新方法。     

血游离DNA检测及肿瘤的基因诊断(文献综述)

肿瘤病人血中有异常升高的游离肿瘤DNA ,用分子生物学方法检测病人血游离DNA中肿瘤相关基因的异常表达 ,是无创伤性诊断肿瘤的新方法     

人胎儿皮肤皮脂腺细胞和外泌汗腺细胞的分离培养及鉴定

目的建立人胎儿皮肤皮脂腺、外泌汗腺细胞的体外分离培养与鉴定方法。方法通过分离人胎儿皮肤皮脂腺腺体和外泌汗腺腺管,以DMEM/F12(1∶1)为基础培养基,分别添加不同浓度的胎牛血清、表皮生长因子、L-谷氨酰胺、氢化可的松、霍乱毒素、青霉素、链霉素、重组人表皮生长因子、三碘甲状腺氨酸、胰岛素、转铁蛋白、亚硒酸钠作为皮脂腺细胞培养基及外泌汗腺细胞培养基,置入37℃、体积分数5%CO2孵箱中进行原代及传代培养。倒置相差显微镜下观察人胎儿皮肤皮脂腺、外泌汗腺细胞的形态及变化,并进行细胞克隆形成率测定。采用油红染色和细胞角蛋白(CK)4.62、上皮膜抗原(EMA)免疫组织化学染色对传代培养的皮脂腺、外泌汗腺细胞进行鉴定。结果分离的人胎儿皮脂腺腺体和外泌汗腺腺管可以在体外贴壁生长繁殖;其皮脂腺细胞的克隆形成率为2.7%,明显低于人胎儿角质形成细胞(8.0%,P<0.01).人外泌汗腺细胞的克隆形成率为7.3%,与人胎儿角质形成细胞(7.7%)比较差异无统计学意义(P>0·05).油红染色显示,皮脂腺细胞含有少量脂质小滴,CK4.62、EMA免疫组织化学染色均为阳性;外泌汗腺细胞CK7、CK19免疫组织化学染色均为阳性。结论用酶消化法和显微分离法可体外分离人胎儿皮肤皮脂腺、外泌汗腺细胞,两者均具备上皮细胞的标志和生物学特点,但皮脂腺细胞增殖速度较为缓慢。     

胎儿宫内窘迫孕妇剖宫产110例分析

目的:探讨胎儿宫内窘迫行剖官产的诊断指标.方法:将110例因胎儿宫内窘迫行剖宫产的孕产妇,按胎心监护情况、羊水情况分为四组,Ⅰ组:为重度胎心异常,Ⅱ组:为轻度胎心异常,Ⅲ组:为轻中度胎心异常伴羊水轻度粪染,Ⅳ组:为单纯羊水Ⅲ粪染,统计其新生儿窒息率的百分比.结果:Ⅰ组:胎儿娩出后新生儿窒息率占47％,81％伴窘迫相关因素,提示胎儿存在缺氧;Ⅱ组:胎儿娩出后阿氏评分均正常,胎窘相关因素少;Ⅳ:组胎儿娩出后,新生儿窒息率占9.1％,相关因素占24.3％.结论:在判断胎儿宫内窘迫时应综合判断,尽可能减少过度诊断,降低剖宫产率.     

产前检查免疫检验项目对孕妇和胎儿的临床价值分析

目的探讨产前检查免疫检验项目对孕妇和胎儿的影响及其临床价值,为临床检验及防治工作提供参考。方法将接受产前检查的1 500例孕妇进行肝炎、梅毒、AIDS、TORCH等免疫检验项目检查,记录并回顾性分析有关资料。结果1 500例受检孕妇中HBs Ag阳性79例(5.27%),HCV阳性4例(0.27%),RPR阳性5例(0.33%),TPPA阳性3例(0.20%),抗-HIV阳性1例(0.07%),抗TORCH-lg M阳性1例(0.07%)。对存在ADIS或对胎儿影响恶劣、难以治愈的病症,取得孕妇及其家属同意后给予终止妊娠。结论产前检查免疫检验项目能够及时发现高危妊娠因素,有利于对孕妇和胎儿采取措施控制,保证其生命安全。     

应用胎儿监护仪对292例孕妇的监测

胎儿监护仪作为监测胎儿宫内窒息参数之一,对早期发现、及时处理、预防新生儿窒息,尤其对高危妊娠、定期观察胎儿在腹中安危,对临床工作有一定指导意义。本文从1988年11月～1989年4月对292例临产的孕妇采用了外监护法,现将监护情况报告如下:     

胎儿异常孕妇自我同情现状及影响因素分析

目的了解胎儿异常孕妇终止妊娠前自我同情现状及其影响因素,为护理人员进行针对性心理干预,改善胎儿异常孕妇心理应激提供依据。方法使用自我同情量表对211例因胎儿异常需引产的孕妇进行调查。结果胎儿异常孕妇自我同情总分为85.19±10.75。多元线性回归结果显示,查出胎儿异常到入院引产时间、不良妊娠史、文化程度是胎儿异常孕妇自我同情的影响因素(P<0.05,P<0.01)。结论胎儿异常孕妇自我同情处于中等水平。护理人员应加强对文化程度低及有不良妊娠史等胎儿异常孕妇的心理疏导,指导其提高自我同情水平,以有效应对心理危机。     

Anesthetic solubility coefficients for maternal and fetal blood.

Solubility coefficients for seven inhalation anesthetic agnets were determined in maternal and fetal blood at 37 C. Halothane, isoflurane, diethyl ether, and nitrous oxide were significantly more soluble in maternal than in fetal blood, while methoxyflurane, fluroxene, and cyclopropane were significantly less soluble. Reasons for these differences cannot be accounted for by differences in the type or amount of hemoglobin present.     

Nitrous oxide concentrations in maternal and fetal blood during caesarean section

BACKGROUND AND OBJECTIVE: There are little data on nitrous oxide (N2O) concentrations in neonatal blood at delivery. We investigated the effects of the time elapsing between the induction of anaesthesia and delivery (the I-D interval) on umbilical blood N2O concentrations. METHODS: Maternal and neonatal blood N2O concentrations were measured in 27 patients undergoing Caesarean section under N2O 67% anaesthesia. The duration of N2O administration (range 2-50 min) was arbitrarily divided into three groups (each n = 9): short (2-9 min), medium (9.1-14 min) and long duration (14.1-50 min). RESULTS: Compared with a rapid increase in the maternal arterial N2O concentration (48.9 +/- 4.7%), the umbilical venous N2O concentration (17.9 +/- 8.3%) rose slowly in the short duration group, whereas the N2O concentrations became more similar (61.6 +/- 4.3 and 43.2 +/- 10.0%, respectively) in the long duration group. The ratio of umbilical vein to maternal artery N2O concentrations correlated with the duration of N2O anaesthesia (r = 0.739), resulting in ratios of 0.37 +/- 0.18, 0.61 +/- 012 and 0.70 +/- 0.13 for the short, medium and long duration groups, respectively. The Apgar score at 1 min correlated inversely with the duration of anaesthesia and with the umbilical vein N2O concentration (r = -0.457 and -0.423, respectively). CONCLUSIONS: The data suggest that placental N2O transfer during Caesarean section is time-dependent and slower compared with maternal N2O uptake. They also suggest that the Apgar score is less affected by N2O administration when the I-D interval is shorter.     

人胚胎神经干细胞的分离培养及鉴定研究

目的：研究人胚胎神经干细胞的培养条件及体外分化情况。方法：从12周龄人胚胎脑皮质分离细胞，采用无血清培养技术，协同应用碱性成纤维细胞生长因子(bFGF)、表皮生长因子(ECF)和重组人白血病抑制因子(rhLIF)进行培养；5-溴脱氧尿嘧啶核苷(BrdU)标记检测细胞的增殖能力，间接免疫荧光化学法检测细胞的分化情况。结果：培养得到的大量半悬浮生长的神经干细胞球能够传代培养；BrdU标记阳性，可诱导分化为神经元、星形胶质细胞和少突胶质细胞。结论：人胚胎脑皮质分离细胞培养得到的细胞群具有神经干细胞的基本特征，可进一步用于基础及临床研究。     

Isolation and characterization of a DNA synthesome from a neuroblastoma cell line

Background

Despite significant analysis of the chromosomal abnormalities associated with neuroblastoma (NB), the role that NB DNA replication may play in the accumulation of genetic damage is poorly understood. For that matter, the mechanisms involved in NB DNA synthesis have yet to be elucidated. In an effort to investigate this process in NB, we have isolated and purified a multiprotein DNA replication complex from human NB cells (IMR-32).

Methods

Using a series of subcellular fractionations, ion-exchange chromatography, and gradient sedimentation steps, we have isolated a simian virus 40 replication competent multiprotein complex from IMR-32 NB cells, which has been designated the DNA synthesome. Enzymatic and immunodetection techniques were used to characterize the multiple components of the multiprotein DNA replication complex.

Results

The NB DNA synthesome was found to remain intact and functional through all the steps of its purification. The proteins and enzymatic activities that were found to copurify with the NB DNA synthesome include: DNA polymerases α, δ, and ?, proliferating cell nuclear antigen, replication factor A, replication factor C, topoisomerases I and II, flap endonuclease 1, and DNA ligase I.

Conclusion

Although the cooperative integration of a DNA replication macromolecular complex (DNA synthesome) is not new, we extend the view of the DNA synthesome mediating DNA synthesis for human NB. The data reported here characterize the human NB DNA synthesome for the first time and provide the groundwork for investigating whether the NB DNA synthesome contributes to faulty DNA replication and tumor pathogenesis for this childhood malignancy.     

孕早期母血胎盘滋养层细胞的检测

为从孕妇血中检测和分离胎盘滋养层细胞，对５５例孕８～１４周母血中细胞滋养层Ｈ３１５和合体滋养层ＦＴ１－４１．１抗原阳性细胞分别进行了免疫细胞化学检测、流式细胞分析、荧光激活细胞分离（ＦＡＣＳ）及荧光原位杂交分析（ＦＩＳＨ）。结果在２８例和１４例孕妇外周血中分别检出了Ｈ３１５和ＦＴ１－４１．１抗原阳性细胞；而１３名怀男胎孕妇外周血Ｈ３１５阳性细胞经ＦＡＣＳ分选及Ｙ染色体探针的ＦＩＳＨ分析，其中杂交阳性细胞含量平均为１１．４±４．３％。认为：孕８～１４周母血循环中存在细胞滋养层和合体滋养层细胞，有可能利用这些细胞在孕早期对胎儿染色体非整倍性异常进行产前分子细胞遗传学诊断。     

Isolation and characterization of a cell line from the epithelial cells of the human fetal pancreas

Pancreatic cell lines are useful for basic studies of pancreatic biology and for possible application to cell transplantation therapies for diabetes. A retroviral vector expressing simian virus 40 (SV40) T antigen and H-ras^va112 was used to infect a monolayer culture of epithelial cells from an 18-wk human fetal pancreas. Infected cells gave rise to a clonal epithelial cell line, designated TRM-1. This cell line expresses epithelial markers as well as glut2 and small amounts of insulin and glucagon. TRM-1 is the first cell line to be generated from the human fetal pancreas and also the first cell line derived directly from the fetal pancreas of any species. The approach that we have used to develop TRM-1 should be applicable to isolating cell lines from other stages of human pancreatic development.     

Trauma in pregnant women: analysis of maternal and fetal mortality.

Twenty-seven traumatised pregnant women were analysed retrospectively over a period of 9 years. Mean age was 23.7 years (16-42 years). Gestational age ranged from 10 to 40 weeks (mean, 21.5 weeks), with most victims (46.1%) being in the second trimester. The predominant mechanism (65.3%) was blunt abdominal injury due to an automobile accident (the patient being run over or collision). At admission, 8 (30.7%) patients had haemodynamic alterations. 6 patients (23.0%) presented vaginal bleeding and 4 of these were haemodynamically normal. We analysed maternal mortality, fetal mortality and their causes. We also compared the median RTS and TRISS values for the groups with maternal-fetal survival and the group with maternal-fetal death. Fetal death occurred in all pregnant women admitted with vaginal bleeding. Maternal mortality due to haemorrhagic shock was 11.5%. Fetal mortality was 30.7%, with 37.5% of these deaths being caused by maternal death. The major cause of fetal mortality was a detached placenta (50.0%). The trauma indices, RTS and TRISS, were significantly lower (p = 0.0025 and p < 0.0001) in the group of maternal-fetal death but they were not of prognostic value in terms of fetal mortality.     

Isolation of osteoclasts from fetal rat long bones

Cell isolates containing multinucleate osteoclasts were obtained from longitudinally split fetal rat long bones by treatment with testicular hyaluronidase. The total yield of osteoclasts and the osteoclast enrichment of the isolate were increased if the intact bones were first cultured for 72 h. Even greater enhancement was obtained if the bones were treated with 1,25-dihydroxycholecalciferol [1,25(OH)2D3] during the culture period. This technique resulted in a cell population containing approximately 15% osteoclasts in yields greater than 50 osteoclasts per long bone. The yield of osteoclasts and the percentage of osteoclasts correlated well with the extent of bone resorption induced by 1,25(OH)2D3. The effectiveness of several isolation procedures was compared using the 1,25(OH)2D3-treated long bones. Conventional digestion with 1 mg/ml crude collagenase gave a much poorer yield of osteoclasts than simply agitating the split long bones. Hyaluronidase plus EDTA was not significantly different from EDTA alone. Even with milder procedures, however, the isolated osteoclasts were damaged as judged by their failure to exclude trypan blue. The osteoclasts are obviously very fragile cells. The isolation technique coupled with May-Grunwald-Giemsa staining permitted reliable determination of the median number of nuclei per osteoclast. This parameter was the same in uncultured bones or in bones cultured for 72 h in control media. Treatment with 1,25(OH)2D3 increased the nuclear number. At lower levels of bone resorption, nuclear number did not increase, but it was significantly greater in more highly resorbed bones.