An improved method of direct chromosome preparation from chorionic villus and high resolution banding technique

The trophoblast was dissociated from the underlying mesenchymal layer either with acetic acid after short-term prefixation or with mechanical power after fixation twice. The colcemid treatment time was shorted to 16 min and trypsin solution of low pH (6.2) was used for banding. By these steps, the quality of chromosome banding was greatly improved and complete standard chromosome diagnoses were made in 24 of 24 cases. With the modified technique, high resolution banding chromosomes were consistently obtained after short-term incubation.     

A simple and reliable method of chromosome banding for prenatal cytogenetics using a bromodeoxyuridine pulse

An extremely simple, fully defined and reliable technique for banding chromosomes in situ is described. The method uses a pulse of bromodeoxyuridine to produce replication pattern banding complete with G- and C-type banding in the same karyotype. This enables immediate resolution of problems otherwise requiring extra subculturing and the application of conventional banding techniques. The value of routine chromosome banding in prenatal cytogenetics using such an economical technique is discussed.     

Synchronization of amniotic fluid cells for high resolution cytogenetics

A high resolution technique was applied to amniotic fluid cells by synchronization. After inoculation, the cells were incubated for 30 h in the presence of either thymidine or 5-bromodeoxyuridine (BrdU). After removal of the blocking agent and addition of a low concentration of thymidine, the cells were incubated for another 6 1/2–7 h, then harvested in prometaphase without colcemid. This technique gives a mitotic index of 3·7 per cent after thymidine synchronization and of 3·2 per cent after BrdU synchronization, and more than half of the mitoses were in the earlier phases with the chromosomes showing more than 550 bands per haploid set. GBG, GTG, and RHG prometaphases are presented. Precise high-resolution banding of chromosomes of amniotic fluid cells can increase diagnostic accuracy.     

A simple method for preparing prometaphase chromosomes from amniotic fluid cell cultures

A simple method for preparing prometaphase chromosomes from amniotic fluid cell cultures is described. The technique is based upon several key steps including: (1) reduced colcemid concentration, (2) reduced exposure to trypsin-EDTA, and (3) maintaining cells in single suspension by adjusting cell concentration appropriately. Chromosomes with banding resolution up ot 800 bands per haploid set can be routinely produced. The described methodology is particularly useful in defining and establishing the clinical significance of subtle structural aberrations.     

Chromosome banding in direct preparations of chorionic villi

Chorionic villus sampling (CVS) is now currently offered for first trimester prenatal diagnosis of genetic disorders. Chromosome analysis of CVS in direct and culture preparations is possible using modifications of standard banding techniques. We summarize our experience in applying QFQ, GTG, RBG, CBG, DA/DAPI, NOR, and SC differentiation protocols to direct preparations. Characteristic chromosome regions are properly labelled by these techniques, and analysis of 300 band stage karyotypes is consistently achievable on GTG banded direct preparations. However, banding of CVS direct chromosomes has proved to be difficult, and the analysis needs to be backed up by culture preparations.     

Correlation between phenotypic expression of de novo marker chromosomes and genomic organization using replicational banding

It has been suggested that actively expressed genes are primarily located in early replicating bands. This hypothesis is supported by cytogenetic and pregnancy outcome data from four consecutive cases of prenatally detected de novo marker chromosomes. Two fetuses with major anomalies had large early replicating bands, while the marker in a third phenotypically normal fetus was late replicating. In the fourth case, a ring marker chromosome had only a small early replicating region. Pregnancy termination was elected. While no structural malformations were apparent, potential intellectual function in this case remains unresolved. An understanding of the relationship between genomic organization and chromosome banding is critical in counseling for prenatally detected de novo marker chromosomes. Replicational banding is particularly helpful in recognizing genes that may be actively expressed and result in developmental abnormality.     

Human chromosomes in prenatal diagnosis: A one step high resolution technique

A simple high resolution technique for human chromosomes is described for fibroblasts obtained from amniotic fluid cell cultures. The application and clinical significance of this technique in prenatal diagnosis is discussed.     

Prenatal diagnosis of a familial complex chromosomal rearrangement involving chromosomes 5, 10, 16 and 18

We report one case of a familial complex chromosomal rearrangement (CCR) involving four different chromosomes 5, 10, 16 and 18. The CCR was detected prenatally at 20 weeks' gestation because of advanced maternal age and history of recurrent miscarriages. Cytogenetic analysis of cultured amniotic fluid cells with GTG banding showed a 46,XX,t(5;16;10;18)(q13;q22;q11.2;q21) karyotype. Parental cytogenetic study revealed that the mother has the same CCR. RBG banding, high-resolution banding and fluorescence in situ hybridization (FISH) were used to characterize further and confirm the conventional banding data. No physical abnormalities were shown in the targeted fetal ultrasonography examination. The parents decided to continue the pregnancy. The child is now 2 years old and has neither congenital anomalies nor evidence of delayed psychomotor development. The fetal targeted ultrasound and FISH analysis helped us reassure fetal status. Copyright © 2002 John Wiley & Sons, Ltd.     

Chorionic villus sampling: Improved method for preparation of karyotypes after short-term incubation

A method for the isolation and trypsin-Giemsa banding of metaphases obtained after short-term incubation (48 h) of cytotrophoblast cells from chorionic villus sample is described. A new slide-making instrument, developed expressly to enhance the spreading of chromosomes from metaphases released from small tissue pieces, is responsible for the increase yield of analysable metaphases in this protocol.     

Prenatal ultrasound diagnosis of macroglossia in the Wiedemann-Beckwith syndrome

We report the ultrasound prenatal diagnosis at the 30th week of macroglossia in two sibs with the Wiedemann-Beekwith syndrome; the syndrome was also present in their mother. A study of high resolution chromosomes did not show any anomaly.     

High resolution chromosomes from first trimester trophoblast cultures

A simple method for obtaining prometaphase chromosomes from cultured first trimester cells involves the addition of BrdU and FdU 11 h before harvest and ethidium bromide 1.5 h before harvest. High resolution R-banding is obtained by acridine orange staining.     

Prenatal foetal diagnosis of partial trisomy 3q and monosomy 13p due to a maternal balanced rearrangement

The authors describe a case of a male foetus whose ultrasound at 20 weeks' gestation revealed cystic hygroma, cleft lip and ventricular septal defect. Amniotic fluid cytogenetics using GTG banding showed a 46,XY,der(13)t(3;13)(q12;p11.1) rearrangement, and fluorescence in situ hybridization (FISH) delineated the relevant breakpoints. Familial studies identified a maternal balanced translocation involving chromosomes 3 and 13. The post-mortem examination confirmed the prenatal ultrasound findings. Copyright © 2005 John Wiley & Sons, Ltd.     

Distal partial trisomy 1q: report of two cases and a review of the literature

We report on two cases with partial trisomy 1q syndrome. One case was a mid-trimester fetus with multiple malformations that was prenatally diagnosed with a de novo distal partial trisomy 1q. Prenatal ultrasound at 24th gestational week demonstrated the presence of cleft lip and palate, increased biparietal diameter and decreased abdominal circumference. Cytogenetic analysis (GTG banding) and subsequent fluorescence in situ hybridization (FISH) using whole chromosome paint 1 and multicolor banding (MCB) demonstrated an aberrant karyotype 46,XY,dup(1)(q31q43∼44). The second case was a newborn male infant with multiple congenital malformations. He had a derivative chromosome 18 as a result of a maternal insertion involving chromosomes 1 and 18. Further analyses including MCB showed his karyotype as 46,XY,ins(18;1)(q22;q23q31.1∼32). The present cases and a review of the literature suggest that partial trisomy of the long arm of chromosome 1 is a distinct clinical entity. Copyright © 2007 John Wiley & Sons, Ltd.     

Outcome of cases of de novo structural rearrangements diagnosed at amniocentesis

The frequency of de novo rearrangements at amniocentesis was determined in 76952 prenatal diagnoses from centres in the United States. Rates for balanced rearrangements are slightly greater than rates previously reported in the newborn, possibly because banding studies were not used in the latter. Rates for unbalanced rearrangements are considerably higher in the amniocentesis data not only because banding was used but also because a substantial loss of abnormal conceptions is to be expected between amniocentesis and birth. The higher frequency of cases with supernumerary markers at amniocentesis is unexplained. A review of 66 apparently balanced de novo rearrangements found at amniocentesis revealed evidence of abnormality in five; in four of these the abnormality was noted in the abortus. The number of cases observed is still too small to rule out a risk of abnormality no greater than the usual rate of abnormalities at birth. Abnormalities were detected in 6 of 10 cases with unbalanced de novo rearrangements. In 33 cases of non-familial supernumerary chromosomes 6 (18.2 per cent) showed abnormality. Non-satellited markers appeared to have a higher rate of abnormality than satellited markers but the difference is not statistically significant. Further studies and improved follow-up of de NOVO cases diagnosed at amniocentesis are required.     

Maternal cell contamination in cultured chorionic villi: Comparison of chromosome Q-polymorphisms derived from villi,fetal skin,and maternal lymphocytes

Maternal cell contamination of chorionic villi (CV) samples used for first trimester prenatal diagnosis can cause obvious and/or unrecognized diagnostic dilemmas. The purpose of this investigation is to assess the frequency of maternal cell contamination (MCC) in chorionic villus samples and to evaluate selected parameters which might predict where contamination is more likely to have occurred. Maternal lymphocytes, chorionic villi from ultrasonically directed transcervical catheter aspiration, and fetal tissue were obtained at 8–11 weeks gestation from 45 patients undergoing elective termination. Quinacrine (Q) banded metaphases were compared from duplicate direct preparations of chorionic villi; cultured chorionic villi, fetal fibroblast tissue cultures, and maternal lymphocyte cultures. Q-polymorphisms in metaphase chromosomes were 100 per cent concordant between fetal tissue and direct CV preparation. However, evidence for maternal cell contamination occurred in 13.1 per cent of cultured chorionic villi preparations where polymorphisms were found to be identical between maternal and cultured CV and both distinct from fetal tissue preparations. Where MCC was identified, it was noted that CV cell cultivation interval was prolonged (24.2±6.8 days) compared with non-contaminated cultures (14.1±4.4 days) (p <0.05). We conclude that maternal cell contamination is a significant problem with chorionic villus sampling. Where direct preparations are not employed or when cultures are ‘slow growing’, MCC may be a significant and unrecognized complication re: fetal diagnosis. Direct preparations, multiple cultures, quinacrine banding, and maternal Q-polymorphism comparisons can minimize diagnostic dilemmas secondary to maternal cell contamination. Q-polymorphism comparisons between maternal and fetal chromosomes should be included in all instances where cultured chorionic villi are utilized for fetal diagnosis and where direct preparations are not available.     

基于MODIS-OLI遥感数据融合技术的农田生产力估算

大范围、高精度的农田生产力遥感监测依赖于高时空分辨率的遥感数据,单纯依靠由单一类型传感器数据获取的高时相或者高空间分辨率的遥感数据都不能满足清晰掌握田块尺度上作物生长动态的需求。全球免费提供的空间分辨率250~1 000 m的MODIS数据和空间分辨率30 m的Landsat数据是植被动态监测普遍应用的数据源,针对应用MODIS数据估算的农田生产力空间分辨率较低而Landsat卫星重访周期长的局限性,研究基于空间分辨率30 m的Landsat 8 OLI数据与空间分辨率500 m的MODIS数据,应用时空数据融合技术,融合OLI数据的高清晰空间表达能力与时间间隔8 d的MODIS数据的植被生长时间序列过程的监测能力,获得空间分辨率30 m、时间步长8 d的时间序列数据,利用VPM （Vegetation Photosynthesis Model）模型以宁夏永宁县部分地区为试验区估算该区域的NPP。研究结果表明,融合后所得30 m分辨率的NPP具有良好的空间细节信息,提高了MODIS数据中混合像元上的估算精度,并保留了MODIS数据原始的时间过程信息,以30 m的空间分辨率刻画出作物的生长动态;较单独应用MODIS数据,使用融合数据估算的NPP可更有效检测出高标准农田建设对农田生产力的提升。     

宁夏银北平原地下水中砷的分布特征及其富集因素

通过对宁夏银北平原地下水中砷的分布特征、来源及富集因素进行研究,结果表明:银北平原地下水砷异常地段的分布具有明显的地带性,从山前洪积倾斜平原前缘向冲湖积平原中心,地下水中砷含量递增,高砷地下水沿黄河河道自南向东北局部呈点状分布,总体上呈条带状展布,且东北部砷含量普遍高于西南部,主要富集在0～40 m深度的含水层中;引黄灌溉的黄河水与贺兰山区的煤系地层是平原内地下水中砷的主要来源;特殊的古地理环境特征、地下水径流条件、氧化还原环境及干旱半干旱的气候条件是地下水中砷富集的重要因素。     

Molecular and cytogenetic characterization of extra-structurally abnormal chromosomes (ESACs) found prenatally: outcome and follow-up

A 40-year-old woman underwent amniocentesis at 15.3 weeks of gestation. Chromosome analysis performed using QFQ, DA-DAPI and CBG banding revealed two de novo extra-chromosomal markers (ESACs) in 11 of the 16 colonies analysed. Fluorescence in situ hybridization (FISH) showed that both chromosomes came from the Yq11.22.1 region of the Y chromosome. PCR analysis of genes and STS localized on the Y chromosome excluded the Yp presence specifically of the SRY gene, and most of the euchromatic region of Yq. After extensive genetic counselling and considering both laboratory and second-level ultrasound data, the couple decided to continue the pregnancy. At 37.4 weeks of gestational age, a girl weighing 2750 g was born with an Apgar score of 9/10. A blood sample taken from the umbilical cord showed three cellular lines:mos47,XX, +mar1 ish.der (Y)(wcpY+) [21%]/48,XX, +mar1 ish.der (Y)(wcpY+), +mar2 ish.der (Y)(wcpY+) [41%]/46,XX [38%]. One year after birth, the baby was developing normally and had normal psychomotorial activity. Copyright © 2003 John Wiley & Sons, Ltd.     

两种典型生活垃圾焚烧炉烟气中二噁(口英)相态分布特征

采集了机械炉捧焚烧炉和循环流化床焚烧炉两种典型生活垃圾焚烧炉排放烟气样品,应用高分辨气相色谱/高分辨质谱(HRGC/HRMS)同位素内标稀释法分别测定了烟气不同相样品中17种2,3,7,8-位氯取代的PCDDs/PCDFs同类物的含量.结果表明,两种炉型中PCDDs/PCDFs同类物及毒性当量贡献率在冷凝水相中所占的比例均在85％以上,远远高于在滤筒相和XAD-2树脂相中所占的比例,机械炉排炉焚烧排放烟气中∑PCDFs与∑PCDDs的比值为0.77;而循环流化床焚烧排放烟气∑PCDFs与∑PCDDs的比值为5.28.机械炉排炉焚烧烟气三相中OCDD为优势分布,尤其是滤简相中OCDD的百分比含量高达51.1％.流化床焚烧炉焚烧烟气滤筒、树脂、冷凝水相中没有出现某个单体对总浓度具有绝对优势的贡献.机械炉排焚烧炉和循环流化床焚烧炉排放的烟气中PCDFs的毒性当量贡献最大,尤其是单体2,3,4,7,8-peCDF对总毒性当量的贡献均在30％以上.     

二噁英类分析用溶剂的超高纯化技术

设计、开发了二英呋喃分析专用高纯溶剂的提纯装置,通过104倍的高度浓缩后,经高分辨率气相色谱-高分辨率质谱联用仪的检测,对该提纯装置的提纯效果进行了验证评价。经验证,国产分析纯级甲苯、正己烷、二氯甲烷、丙酮等溶剂中二英呋喃类以及其他杂质等干扰物的存在量被控制在10-15g以下(仪器检测下限值)。该溶剂提纯装置体积小、操作方便,能够满足二英研究实验室所需的超高纯品质的二英级溶剂的需求,极大地降低了二英的研究成本。