Re-evaluation of the colorimetric assay for cytidine deaminase activity

This study re-evaluated the colorimetric assay for cytidine deaminase (CTD), and showed that the optimum conditions were pH 7·5, 37°C, and up to 24 h. In addition, this method was found to require protein precipitation. Following these modifications, intra-assay and inter-assay coefficients of variation were below 5 per cent, indicating that the assay was highly reliable. CTD activity was determined in 282 serum samples from 206 normal pregnant women by the incubation of 100 μl of serum and 400 μl of 1·4 mmol/l cytidine substrate for 16 h at 37°C. Following protein precipitation, the ammonia liberated during conversion was measured by a colorimetric procedure. The mean (±SD) CTD activity was 7·31 ± 2·50 U at 3–12 weeks of gestation, 8·70 ± 2·12 U at 13–24 weeks, 7·59 ± 2·25 U at 25–36 weeks, and 7·29 ± 2·16 U at 37–42 weeks. High levels of CTD activity were found in patients with abruptio placentae and amnionitis associated with intrauterine fetal death (IUFD). The increase in CTD activity was noted from 3 days to 1 week before the confirmation of IUFD. The placenta contains extremely high levels of CTD, but cord serum does not. Thus, the excessive elevation of CTD activity was probably derived from progressive placental damage. This modified CTD assay was concluded to be simple and reliable, and may perhaps be useful in detecting pregnancy disorders.     

Human chorionic gonadotropin and unconjugated oestriol measurements in insulin-dependent diabetic pregnant women being screened for fetal down syndrome

This prospective study investigates the relationship between insulin-dependent diabetes and maternal serum levels of alpha-fetoprotein (AFP), unconjugated oestriol (uE3), and human chorionic gonadotropin (hCG). It also examines the potential impact on screening for Down syndrome. The population-based cohort included 20 321 pregnant women in Maine who underwent routine serum screening for Down syndrome in the second trimester. The cohort included 52 women with insulin-dependent diabetes. Maternal serum AFP levels are now routinely adjusted for insulin-dependent diabetes. These adjustments, therefore, were made routinely in the diabetic women, but no equivalent adjustments were made for uE3 and hCG values. The initial false-positive rate (using all three markers) among the women with diabetes was not significantly different from that in the non-diabetic population (7·7 and 5·4 per cent, respectively). Prior to adjustment for insulin-dependent diabetes, the median AFP level in the 52 women was 0·73 multiples of the median (MOM); the median levels of uE3 and hCG were 0·93 and 0·98 MOM, respectively. When the uE3 and hCG levels were adjusted, the initial false-positive rate was unchanged. Median serum levels of uE3 were significantly higher in the 33 women whose onset of diabetes was prior to 19 years of age (0·99 MOM) than in the 19 women whose onset of diabetes was at age 19 or older (0·84 MOM). This is the first population-based study to investigate the relationship between diabetes and serum levels of AFP, uE3, and hCG, and confirms earlier observations from a case—control study that found only slightly lower uE3 and hCG levels.     

Blood transferrin receptor expression in chromosomally abnormal fetuses

In a cross-sectional study of 13 chromosomally abnormal fetuses, umbilical venous blood was obtained by cordocentesis at 17–32 weeks' gestation. Fetal blood transferrin receptor (CD71) expression (mean=79·8 per cent, range=60–98 per cent) and nucleated red cell count (mean=10·4 × 10⁹ per 1, range=1·0–25·0 × 10⁹ per 1) were significantly higher than the appropriate normal mean for gestation (z=3·92, P<0·0001 and z=3·69, P<0·001, respectively). These haematological changes in chromosomally abnormal fetuses would facilitate their prenatal diagnosis by analysis of fetal nucleated red blood cells isolated from the maternal circulation on the basis of CD71 expression.     

1株分离自煤矿废水的铁硫氧化细菌LY01的鉴定及其氧化特性研究

从贵州省某煤矿废水中分离出1株嗜酸性的铁硫氧化细菌菌株LY01,经过对该菌株的形态特征、理化特征检测及16S rRNA序列分析,初步鉴定菌株LY01为嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans).在无机盐培养液中菌株LY01可以从氧化Fe2+、单质硫(S0)及黄铁矿(FeS2)获得能量生长,对Fe2+的氧化速率最快,反应30 h即可将44.2 g.L-1的FeSO4.7H2O完全氧化,菌体细胞达到4.2×107个.mL-1;反应21 d可将溶液中10 g.L-1含量的S0氧化6.7%,溶液中SO24-浓度达2 001 mg.L-1,菌体细胞达到8.9×107个.mL-1;反应20 d可将溶液中30 g.L-1含量的FeS2氧化10%,溶液中SO24-浓度达4 443 mg.L-1,菌体细胞达到3.4×108个.mL-1.研究重金属离子Ni2+、Pb2+对菌株LY01氧化FeS2活性的影响,结果发现10～100 mg.L-1的Ni2+对菌株LY01氧化FeS2活性有一定的影响,而10～100 mg.L-1Pb2+对菌株LY01氧化FeS2的活性无影响.     

Serum hCG assay: A method for detection of contamination of fetal blood samples

Serum human chorionic gonadotrophin (hCG) can be assayed in specimens obtained by percutaneous fetal blood sampling to check for the absence of maternal blood or amniotic fluid contamination. In order to assess the accuracy of this approach, we measured serum hCG in 44 pure fetal blood samples obtained by intracardiac puncture. The mean fetal serum hCG concentration was 52 IU/l, and the ratio of maternal to fetal serum hCG concentration never exceeded 1 · 1 per cent, which represents the smallest contamination rate detectable by this method.     

Maternal serum free beta hCG screening: Results of studies including 480 cases of down syndrome

The median maternal serum free beta human chorionic gonadotropin (hCG) multiple of the median (MOM) of 480 Down syndrome cases in the second trimester was 2·64, significantly greater than the reported median MOM of intact hCG (p<0·0001). In 234 of these cases from retrospective and prospective studies, the effectiveness of maternal serum free beta hCG was evaluated in combination with alpha-fetoprotein (AFP) and maternal age in second-trimester Down syndrome screening. Down syndrome detection in the gestational age range of 14–16 weeks was 82 per cent. In all gestational weeks (14–22), a 77·7 per cent Down syndrome detection rate was achieved. In prospective screening of 44 272 patients under the age of 35 years, 69 per cent of Down syndrome cases were detected (73 per cent in gestational weeks 14–16). The false-positive rate for the prospective study was 3·8 per cent. The use of free beta hCG combined with maternal serum AFP and maternal age-related risk for Down syndrome in a screening population (i.e., women under 35 years) yields an improved detection efficiency over other protocols.     

The influence of serum screening on the amniocentesis rate in women of advanced maternal age

We investigated the effect of maternal serum screening on the amniocentesis (AC) rate in women of advanced maternal age. The AC rate after maternal serum screening was compared in two groups of women with a singleton pregnancy, 855 women of 30–35 years and 98 of 36 years and older. In our population, 34·1 per cent of the women of 36 years or older were ‘screen-positive’ for Down syndrome. Only 41·2 per cent of these women chose to undergo AC as opposed to 88·2 per cent in the younger age group. Within the older age group, the tendency to avoid AC increased with increasing age. Maternal serum screening led to a significant decrease in the AC rate in the older women. In this group, a comparison between the ‘a priori’ and the calculated risk might have had more influence on the decision to undergo AC than being screen-positive or screen-negative as such. We conclude that maternal serum screening had a major effect on the AC rate in women of advanced maternal age. This is of importance in a society in which the average maternal age is steadily increasing.     

2,3,7,8,-四氯苯噁英对NIH小鼠早期妊娠和生殖激素的影响

用不同剂量的2,3,7,8 四氯苯二英(2,3,7,8 tetrachlorodibenzopdioxin,TCDD)对小鼠胚胎附植数量和生长发育情况进行剂量反应评估,分析TCDD对假孕小鼠子宫蜕膜生长状况的影响以及比较胚胎种植前后染毒效果.所有样本采集血液,测定母体血清雌二醇和孕酮的浓度变化.结果表明,50和100ng·(kg·d)-1TCDD处理的孕鼠,胚胎附植数量明显减少(P<0 05),着床后胚胎出现发育迟缓,质量减轻(P<0 05);同等剂量TCDD处理的假孕小鼠,子宫蜕膜生长受到了极显著的抑制(P<0 01);种植前后染毒效果比较发现种植前期染毒的小鼠胚胎对TCDD更为敏感.母体血清雌二醇浓度随剂量和染毒时段有不同程度的升高,而孕酮浓度则极显著下降(P<0 01).由此表明低剂量TCDD可能通过对生殖激素的干扰,强烈影响NIH小鼠妊娠早期生殖生理状态和着床后胚胎的发育.实验同时证明小鼠也可作为早期妊娠毒物研究的供试动物,用以筛选动物和人类早期妊娠的有害化合物.     

Preparation of hapten-specific monoclonal antibody for cadmium and its ELISA application to aqueous samples

High-affinity and specific monoclonal antibodies against cadmium-ethylene diamine tetraacetic acid (EDTA) complex have been produced using the hybridoma technique. A hapten was synthesized and characterized by Fourier Transform Infrared Spectroscopy (FT-IR) and UV-Vis. Competitive enzyme-linked immunosorbent assay (ELISA) for quantitative detection of cadmium in aqueous sample was developed. The monoclonal antibody with high level of binding affinity for Cd-IEDTA-BSA and high specificity for soluble Cd-EDTA complex showed less than 0.99% cross-reactivity with other 11 metals. The limit of detection was 0.10 μg·L⁻¹, and the effective linear range was 10⁻¹–10³ μg·L⁻¹. The intra- and inter-assay coefficient variations were 1.5%–6.3% and 3.2%–7.4%, respectively. The spike recovery in different water samples were between 98.5% and 110.3%. The detection limit of this assay was well below the allowable concentration of cadmium (3 μg·L⁻¹), and the working range was wider than that of other methods which showed the range of 2.19–86.38 and 0–10³ μg·L⁻¹. The competitive ELISA established in this paper was sensitive and accurate in the screening of cadmium in aqueous samples. The results will lay a solid foundation for construction of an immunoassay kit for cadmium.     

Prenatal diagnosis of glutathione synthase deficiency

Prenatal diagnosis for glutathione synthase (EC 6·3.2·3) deficiency in two pregnancies of an at-risk couple was performed on amniotic fluid taken at 16 weeks' gestation. 5-Oxoproline (pyroglutamic acid) levels were 970 and 790 μmol/l compared with the normal mean value of 29 μmol/l (range 13–51 μmol/l). The pregnancies were terminated and the diagnosis in one case was subsequently confirmed by assay of glutathione synthase in cultured fetal fibroblasts. In the other, post-mortem tissue samples failed to grow.     

Maternal serum free α-human chorionic gonadotrophin levels in twin pregnancies: Implications for screening for Down's syndrome

Maternal serum free α-human chorionic gonadotrophin (free α-hCG) levels were determined in twin and singleton pregnancies at 15–22 weeks of gestation using a set of stored serum samples relating to 200 twin pregnancies and 600 singleton control pregnancies matched for gestational age and duration of storage. Free α-hCG values are, on average, 1·66 times greater in twin pregnancies than in singleton pregnancies (95 per cent confidence interval 1·56–1·76). If maternal serum free α-hCG is used in screening for Down's syndrome, values in twin pregnancies can be adjusted using this result so that screening can be performed in twin pregnancies as well as in singleton pregnancies.     

Amniotic fluid acetylcholinesterase measurements: Comparing immunochemical and polyacrylamide gel techniques

In the present study, a recently reported immunochemical technique for measuring acetylcholinesterase (AChE) in amniotic fluid utilizing the 4F19 antibody was compared with the widely utilized polyacrylamide gel technique to determine whether the immunochemical assay provided an advantage in separating unaffected pregnancies from those associated with open spina bifida (OSB) and open ventral wall defects (OVWD). The study included (1) 73 amniotic fluid samples from unaffected pregnancies [alpha-fetoprotein (AFP) < 2 MoM] with no visible gel AChE band, (2) nine bloodstained samples from unaffected pregnancies (AFP 2·2–4·0 MoM) with visible gel AChE bands, (3) 18 samples associated with OSB (AFP 2·2–7·0 MoM) with visible gel AChE bands, and (4) 20 samples associated with OVWD (AFP 3·2–53·5 MoM) with visible gel AChE bands. The immunochemical assay produced ranges of measurements in the four respective categories as follows: (1) 2–60 arbitrary units (AU): (2) 14–69 AU, (3) 61–593 AU, and (4)22–476 AU. Eight of the nine unaffected pregnancies with visible gel AChE bands had immunochemical measurements below the highest measurement for the samples with no visible AChE band (60 AU), as did five out of 20 OVWD pregnancies. Two of the OSB cases had values of 61 and 62 AU. These data indicate that the 4F19 specific monoclonal antibody to AChE is capable of distinguishing unaffected from affected pregnancies with reasonable reliability but that more work needs to be done to establish the extent of overlap between the unaffected and affected populations.     

辽河口湿地沉积物硝化细菌及硝化作用研究

2009年6月和8月,采用现场培养和实验室模拟培养相结合的方法对辽河口湿地表层沉积物硝化细菌数量、硝化速率及影响因素进行了研究.结果表明,辽河口湿地表层沉积物氨氧化细菌(ammonia-oxidizing bacteria,AOB)数量6月在0.54×104～5.69×104个.g-1之间,平均值为(2.21±2.32)×104个.g-1,8月在1.90×104～7.90×104个.g-1之间,平均值为(3.61±2.87)×104个.g-1;沉积物潜在硝化速率6月在9.72～16.45 mmol.(m2.h)-1之间,平均值为(12.54±3.14)mmol.(m2.h)-1,8月在14.66～24.62 mmol.(m2.h)-1之间,平均值为(18.71±4.21)mmol.(m2.h)-1;净硝化作用速率6月(S1站)为0.41 mmol.(m2.h)-1,8月在0.20～0.53 mmol.(m2.h)-1之间,平均值为(0.35±0.16)mmol.(m2.h)-1.潜在硝化速率显著高于净硝化速率,AOB数量、净硝化作用速率和潜在硝化作用速率均表现为8月高于6月,芦苇根际效应对硝化作用有促进作用.通过SPSS 13.0软件统计分析,表明影响辽河口湿地表层沉积物硝化作用的主要环境因子有上覆水NH 4+-N浓度和沉积物pH、有机质、总氮(TN)、总磷(TP)、NH 4+-N含量以及AOB数量(p0.05),其中上覆水NH 4+-N浓度和沉积物总磷(TP)、NH 4+-N含量对硝化作用影响较大,是辽河口湿地硝化作用影响的关键因素.根据研究结果估算辽河口湿地沉积物硝化作用每天可以将1.14×105kg的NH 4+-N转化为NO 3--N,对河口湿地氮的循环具有重要意义.     

莠去津降解菌HB-5的最佳产酶培养基及发酵条件

从农药厂废水中分离到一株降解莠去滓的节杆菌(Arthrobacter sp.)HB-5,以从该菌中提取到的降解酶对莠去津的降解率为指标,进行最佳产酶培养基及发酵条件的优化研究,对其产酶量进行了评价.通过正交试验和均匀试验,对细菌HB-5的发酵培养基进行了优化研究.运用SAS软件进行结果分析,所获优化培养基配方为:蔗糖3.0g·L-1,莠去津0.38g·L-1,K2HPO40.5g·L-1,KH2PO41.2 g·L-1,MgSO4·7H2O 1.2g·L-1,NaCl 0.1g·L-1,微量元素溶液3.8mL·L-1.得到菌株培养的最佳优化条件为:菌株发酵液培养时间为48h.接种量为2%,发酵液初始pH值为9,250mL三角瓶中装液量为80mL经优化后,降解酶对莠去津的降解率(91.64%)比原培养基(40.67%)提高了125%.     

OH自由基绝对标定系统的建立和研究

利用H2 O在 1 85nm紫外光照射下产生定量OH自由基 ,用低压扩散激光诱导荧光技术建立了OH自由基标定系统 .在实验条件下 ,体系产生OH自由基的浓度范围为 3× 1 0 7— 2× 1 0 9个·cm- 3,得到归一化荧光信号 (S)与OH自由基浓度的定量关系为S =6 0 4× 1 0 - 9[OH]- 0 0 4 0 7(R2 =0 987) ,并研究和讨论了饱和效应以及体系中可能存在的干扰 .     

基于沼液的培养基及产油小球藻藻种选育

将产油小球藻培养与沼液污水处理结合,为小球藻生长提供营养和水源,同时实现了沼液污水的无害化处理.本研究利用4种产油小球藻,在沼液污水与绿藻培养基体积比为1∶9、1∶3、1∶1、3∶1形成的培养基中培养,以产油率为指标,选育出获得最高产油率时的沼液污水-绿藻培养基配比和小球藻藻种.结果表明,产油率最高的培养基体积比为1∶3,小球藻藻种为BJ05,该工况下产油率达到9.20 mg·(L·d)-1,高于纯绿藻培养基中的8.66 mg·(L·d)-1.在1/4污水比例培养基基础上,考查添加绿藻培养基中不同营养组分对BJ05产油率的影响,结果发现,在同时不添加碳酸钠和柠檬酸的情况下,BJ05的产油率为9.36 mg·(L·d)-1,COD、TN(总氮)、TP(总磷)、NH+4-N去除率分别达到59%、75%、61%、100%.而其他营养成分缺失则显著降低了BJ05的生物量,进而降低了产油率,所以进一步优化培养基为绿藻培养基中不添加碳酸钠和柠檬酸的体积比为1∶3的沼液污水-绿藻培养基.     

First-trimester maternal serum schwangerschafts protein 1 (SP1) in pregnancies associated with chromosomal anomalies

The relationship between first-trimester maternal serum Schwangerschafts protein 1 (SP1) and the karyotype of the pregnancy was examined in 692 women who underwent chorionic villus biopsy at 6–12 weeks. There were 30 pregnancies with abnormal karyotypes, consisting of 14 Down's syndrome (DS), eight trisomy 18, and eight other anomalies, two of which were mosaics. The normal ranges and medians for gestation were defined from the 662 cases in which the karyotype was normal. The median SP1 (0·5 MOM) of the abnormal group was significantly lower than that of the normal group (10 MOM). This relationship was maintained for the DS pregnancies (0·4 MOM) and for anomalies other than trisomy 18 (0·43 MOM) but not trisomy 18 (1·1 MOM). It is possible that the use of SP1 as a screening test for chromosome anomalies in the first trimester could have a 43 per cent detection rate for a 5 per cent false-positive rate.     

MSAFP levels and oesophageal atresia

This study was undertaken to evaluate the relationship between maternal serum alpha-fetoprotein (MSAFP) levels and oesophageal atresia (OA). OA occurred in 16 fetuses of mothers who had an MSAFP test in the study interval. The multiple of the median (MOM) value for MSAFP averaged 1·54 ± 0·65 (range 0·5–2·9 MOM), which was significantly higher than the value seen in controls. The median MOM was 1·35. Using a cut-off of 2·5 MOM, the sensitivity of MSAFP for detecting OA was 19 per cent. Although OA should be considered in the differential diagnosis of an elevated MSAFP level, MSAFP cannot be considered an appropriate screening test for OA given the low sensitivity.     

Amniotic fluid-AFP in Down syndrome and other chromosome abnormalities

80·2 Per cent of 111 Down syndrome pregnancies had amniotic fluid (AF) alpha fetoprotein (AFP) levels on or below the median and 10·8 per cent at or below 0·5 MoM compared with 41·9 and 1·4 per cent of controls. These differences were even more striking when the gestational age was < 18 weeks compared with ⩾ 18 weeks. No such association was seen for other chromosome abnormalities including trisomy 18,45,X and mosaics, 47,XXY, 47,XXX, and other structural abnormalities and triploidy, even when high levels due to defects such as omphalocele and cystic hygroma were excluded. All cases of trisomy 13 and 80 per cent with 47,XYY had AF-AFP levels above the median. Selection of cases for karyotyping by a low level of AF-AFP would clearly fail to detect aneuploidies other than Down syndrome and is not recommended. A possible weak association between low maternal serum (MS) and AF-AFPs in Down syndrome was most evident at < 18 weeks, suggesting that MS screening between 16 and 18 weeks may be the most informative time.     

对-偶氮苯重氮氨基偶氮苯磺酸测定水中痕量汞的研究

本文报导一种高灵敏度、高选择性、水相直接显色测定痕量汞的新方法。在Triton X-100存在下,NH₃·H20-NH₄Cl体系中,汞与新显色剂对-偶氮苯重氮氨基偶氮苯磺酸(ADAAS)有灵敏的显色反应。红色络合物在528nm处摩尔吸光系数为1.76×105L·mol-1cm-1。金属离子中仅镉有干扰,可用氨三乙酸掩蔽。