Compound heterozygous B3GALNT2 mutations in a fetus with encephalocele: A case report.

An encephalocele is a congenital malformation characterized by protrusion of the intracranial contents through a cranial defect. We report that a fetus of a pregnant mother who had two consecutive pregnancies with ultrasound-detected encephalocele carried compound heterozygous variants in B3GALNT2 NM_152490.5:c.[1423C > T (p.Gln475Ter)]; [261-2A > G] of maternal and paternal origins, respectively, as confirmed by exome sequencing followed by Sanger sequencing validation. The present case implies that mutations in B3GALNT2, a well-known dystroglycanopathy causative gene, may result in a phenotype of neural tube defect, providing new insights into the clinical spectrum of B3GALNT2-related disorders. Our study may contribute to prenatal screening/diagnosis and genetic counseling of congenital brain malformations.

Maternal Outcomes Among Pregnant Women With Congenital Heart Disease-Associated Pulmonary Hypertension.

BACKGROUND: Studies focused on pregnant women with congenital heart disease (CHD)-associated pulmonary hypertension (PH) are scarce and limited by small sample sizes and single-center design. This study sought to describe the pregnancy outcomes in women with CHD with and without PH. METHODS: Outcomes for pregnant women with CHD were evaluated retrospectively from 1993 to 2016 and prospectively from 2017 to 2019 from 7 tertiary hospitals. PH was diagnosed on the basis of echocardiogram or catheterization. The incidence of maternal death, cardiac complications, and obstetric and offspring complications was compared for women with CHD and no PH, mild, and moderate-to-severe PH. RESULTS: A total of 2220 pregnant women with CHD had completed pregnancies. PH associated with CHD was identified in 729 women, including 398 with mild PH (right ventricle to right atrium gradient 30-50 mm Hg) and 331 with moderate-to-severe PH (right ventricle to right atrium gradient >50 mm Hg). Maternal mortality occurred in 1 (0.1%), 0, and 19 (5.7%) women with CHD and no, mild, or moderate-to-severe PH, respectively. Of the 729 patients with PH, 619 (85%) had CHD-associated pulmonary arterial hypertension, and 110 (15%) had other forms of PH. Overall, patients with mild PH had better maternal outcomes than those with moderate-to-severe PH, including the incidence of maternal mortality or heart failure (7.8% versus 39.6%; P<0.001), other cardiac complications (9.0% versus 32.3%; P<0.001), and obstetric complications (5.3% versus 15.7%; P<0.001). Brain natriuretic peptide >100 ng/L (odds ratio, 1.9 [95% CI, 1.0-3.4], P=0.04) and New York Heart Association class III to IV (odds ratio, 2.9 [95% CI, 1.6-5.3], P<0.001) were independently associated with adverse maternal cardiac events in pregnancy with PH, whereas follow-up with a multidisciplinary team (odds ratio, 0.4 [95% CI, 0.2-0.6], P<0.001) and strict antenatal supervision (odds ratio, 0.5 [95% CI, 0.3-0.7], P=0.001) were protective. CONCLUSIONS: Women with CHD-associated mild PH appear to have better outcomes compared with women with CHD-associated moderate-to-severe PH, and with event rates similar for most outcomes with women with CHD and no PH. Multimodality risk assessment, including PH severity, brain natriuretic peptide level, and New York Heart Association class, may be useful in risk stratification in pregnancy with PH. Follow-up with a multidisciplinary team and strict antenatal supervision during pregnancy may also help to mitigate the risk of adverse maternal cardiac events.

Perinatal maternal characteristics predict a high risk of neonatal asphyxia: A multi-center retrospective cohort study in China.

Objective: This study aimed to identify various perinatal maternal characteristics that contributed to neonatal asphyxia (NA) in term and late-preterm newborns based on the data obtained from a Chinese birth registry cohort and to establish an effective model for predicting a high risk of asphyxia. Method: We retrospectively reviewed and analyzed the birth database from July 1, 2016, to June 30, 2017, in the main economically developed regions of China. Asphyxia was defined as an Apgar score <7 at 5 min post-delivery with umbilical cord arterial blood pH < 7.2 in the infant born after 34weeks. We compared the perinatal maternal characteristics of the newborns who developed asphyxia (NA group, n = 1,152) and those who did not (no NA group, n = 86,393). Candidate predictors of NA were analyzed using multivariable logistic regression. Subsequently, a prediction model was developed and validated by an independent test group. Result: Of the maternal characteristics, duration of PROM ≥ 48 h, a gestational week at birth <37, prolonged duration of labor, hypertensive disorder, nuchal cord, and birth weight <2,500 or ≥4,000 g, abnormal fetal heart rate, meconium-stained amniotic fluid, and placenta previa were included in the predicting model, which presented a good performance in external validation (c-statistic of 0.731). Conclusion: Our model relied heavily on clinical predictors that may be determined before or during birth, and pregnant women at high risk of NA might be recognized earlier in pregnancy and childbirth using this methodology, allowing them to avoid being neglected and delayed. Future studies should be conducted to assess its usefulness.

Heat Transfer Enhancement of n-Type Organic Semiconductors by an Insulator Blend Approach.

The transfer of heat energy in organic semiconductors (OSCs) plays an important role in advancing the applications of organic electronics, especially for lifetime issues. However, compared with crystalline inorganic semiconductors, the thermal transport of OSCs is less efficient and a relevant understanding is very limited. In this contribution, we show that the heat conduction of OSCs can be enhanced by blending with a "commodity" insulator (both thermal and electrical). PC71BM, a well-known electron transporter but poor thermal conductor, was selected as the host OSC material. The blending of a small amount of polystyrene (PS), a commonly used insulating polymer, can facilitate the heat transfer of PC71BM films, as substantiated by the scanning photothermal deflection technique and an infrared thermal camera. The phase thermodynamics of PC71BM/PS blends indicates that the efficient heat transfer preferably occurs in the OSC/insulator blends with better intimate mixing, where isolated PC71BM domains can be effectively bridged by PS that thread through the regions. The applicability of this approach can be observed in blends with another host material─ITIC. This work provides a facile strategy for designing thermally durable organic electronic devices.

Molecular Doping Increases the Semitransparent Photovoltaic Performance of Dilute Bulk Heterojunction Film with Discontinuous Polymer Donor Networks.

The semitransparent and colorful properties of organic solar cells (OSCs) attract intensive academic interests due to their potential application in building integrated photovoltaics, wearable electronics, and so forth. The most straightforward and effective method to tune these optical properties is varying the componential ratio in the blend film. However, the increase in device transmittance inevitably sacrifices the photovoltaic performance because of severe carrier recombination that originates from discontinuous charge-transport networks in the blend film. Herein, a strategy is proposed via the molecular-doping strategy to overcome these shortcomings. It is discovered that p-doping is able to release the trapped holes in segregated polymer domains leading to short-circuit current enhancement, while n-doping is more effective to fill the bandgap states producing a higher fill factor. More importantly, either type of doping improves the photovoltaic performance in the semitransparent photovoltaic devices. These discoveries provide a new pathway to breaking the compromise between the photovoltaic performance and optical transmittance in semitransparent OSCs, and hold promise for their future commercialization.

Incidence and Risk Factors of Postpartum Hemorrhage in China: A Multicenter Retrospective Study.

Background: Postpartum hemorrhage (PPH) is a leading cause of maternal morbidity and mortality worldwide but the incidence and its risk factors in China is limited. The objective of this study is to investigate the incidence and the risk factors of PPH in Chinese women. Methods: A multi-center retrospective study of pregnant women at ≥28 weeks of gestation was conducted. Logistic regression was used to identify potential risk factors of PPH and receiver operating characteristic curve was used to evaluate the predictive performance of the identified risk factors. Subgroup analysis focusing on the number of fetus and the mode of delivery was conducted. Results: A total of 99,253 pregnant women were enrolled and 804 (0.81%) experienced PPH. The subgroup analysis revealed that the incidence of PPH was 0.75, 2.65, 1.40, and 0.31% in singletons, twin pregnancies, cesarean sections, and vaginal deliveries, respectively. Placenta previa and placenta accreta were the predominant risk factors of PPH in the overall population and all subgroups. A twin pregnancy was a risk factor for PPH regardless of the mode of delivery. Obesity, and multiparity were risk factors for PPH in both singletons and cesarean section cases, but the latter predicted a reduced probability of PPH in vaginal deliveries. Macrosomia was associated with increased risk of PPH in singletons or vaginal deliveries. In women who delivered vaginally, preeclampsia was associated with a higher risk of PPH. The areas under the curve for the overall cohort, singletons, twin pregnancies, cesarean section cases, and vaginal deliveries were 0.832 (95% confidence interval [CI] 0.813-0.851), 0.824 (95% CI 0.803-0.845), 0.686 (95% CI 0.617-0.755), 0.854 (95% CI 0.834-0.874), and 0.690 (95% CI 0.646-0.735), respectively. Conclusions: The risk factors of PPH varied slightly based on the number of fetuses and the mode of delivery, while placenta previa and placenta accreta were the two major risk factors. A combination of the identified risk factors yielded a satisfactory predictive performance in determining PPH in the overall cohort, singletons pregnancies, and women who delivered by cesarean section, whereas the performance was moderate in twin pregnancies and in women delivering vaginally.

Perinatal Outcomes and Risk Factors for Preterm Birth in Twin Pregnancies in a Chinese Population: A Multi-center Retrospective Study.

Background: Twin pregnancies are associated with an increased risk of adverse maternal and neonatal outcomes, mainly owing to prematurity. Few studies have evaluated the risk factors for preterm birth (PTB) in Chinese population. The objective of this study is to present the short-term maternal-neonatal outcomes, investigating the potential risk factors associated with preterm birth in Chinese twin pregnancies. Methods: A multi-center retrospective study of women pregnant with twins ≥28 weeks of gestation was conducted. Maternal and neonatal outcomes were analyzed. Logistic regression was used to identify potential risk factors for PTB before 37, 34, and 32 weeks, respectively. Results: A total of 3,288 twin pregnancies and 6,576 neonates were included in 99,585 pregnancies. The rate of twin pregnancy was 3.3%, while the PTB rate before 37, 34, and 32 weeks among this population were 62.1, 18.8, and 10.4%, respectively. Logistic regression revealed that monochorionicity [Odds ratio (OR) 3.028, 95% confident interval (CI) 2.489-3.683, P < 0.001], gestational weight gain (GWG) <10 kg (OR 2.285, 95% CI 1.563-3.339, P < 0.001) and GWG between 10 and 15 kg (OR 1.478, 95% CI 1.188-1.839, P < 0.001), preeclampsia (PE) (OR 3.067, 95% CI 2.142-4.390, P < 0.001), and intrahepatic cholestasis of pregnancy (ICP) (OR 3.122, 95% CI 2.121-4.596, P < 0.001) were the risk factors for PTB before 37 weeks. Monochorionicity (OR 2.865, 95% CI 2.344-3.501, P < 0.001), age < 25 years (OR 1.888, 95% CI 1.307-2.728, P = 0.001), and GWG <10 kg (OR 3.100, 95% CI 2.198-4.372, P < 0.001) were risk factors for PTB before 34 weeks. Monochorionicity (OR 2.566, 95% CI 1.991-3.307, P < 0.001), age younger than 25 years (OR 1.964, 95% CI 1.265-3.048, P = 0.003), and GWG <10 kg (OR 4.319, 95% CI 2.931-6.364, P < 0.001) were the risk factors for PTB before 32 weeks. Conclusions: Monochorionicity and GWG <10 kg were two major risk factors for PTB before 32, 34, and 37 weeks, whereas maternal age, PE, and ICP were also risk factors for PTB in specific gestational age.

Analysis of clinical features of 231 cases with pernicious placenta previa: A retrospective cohort study.

ABSTRACT: Pernicious placenta previa (PEPP) is a severe complication of late pregnancy, which might result in adverse maternal-fetal outcome. To explore the application value of placenta accreta score (PAS) for PEPP and its association with maternal-fetal outcome.In this retrospective cohort study, the clinical data of PEPP patients were analyzed. According to the ultrasonic PAS, patients were grouped into 3 groups: scores ≤5, a scores between 6 and 9, and scores ≥10. The clinical data, intraoperative and postoperative outcomes were collected. Receiver operating characteristic (ROC) curves were used to evaluate the performance of PAS in disease severity evaluation. Multivariate logistic and linear regression analysis were performed to assess associations of PAS with intraoperative and postoperative outcomes.A total of 231 patients were enrolled. There were significant differences in intraoperative, postoperative and neonatal outcomes, such as operation time, bladder repair, ICU admission, postoperative hospitalization days, operation complications, Apgar score of newborns in 1 minute and premature delivery among the 3 groups (all P < .05), while the worst outcomes were found in those with a score ≥ 10 (all P < .05). According to ROC curves, scores <5.5, between 5.5 and 7.5, and >7.5 indicated placenta accreta, placenta increta and placenta percreta, respectively. PAS was independently associated with longer time of operation, surgical complications, intraoperative bleeding volume, and postoperative hospitalization days (all P < .05).Placenta accreta score might help with PEPP subtype diagnosis and predict the maternal-fetal outcome of PEPP patients.

Obstetric outcomes for twins from different conception methods - A multicenter cross-sectional study from China.

INTRODUCTION: The effects of assisted reproductive technology on the outcomes of twin pregnancies are controversial. Therefore, the purpose of this study was to compare the maternal and perinatal outcomes of twin pregnancies conceived spontaneously and those conceived by assisted reproductive technology. MATERIAL AND METHODS: This was a cross-sectional study performed at Peking Union Medical College Hospital (PUMCH). Data on twin pregnancies (conceived spontaneously and by in vitro fertilization [IVF]/intracytoplasmic sperm injection [ICSI]) were obtained from the National Birth Registry of China for the period between 1 October 2016, and 30 September 2017. The primary obstetric outcomes were compared between twin pregnancies conceived by different methods. Logistic regression analysis with 95% confidence intervals (95% CI) was used for the multivariate analysis. RESULTS: A total of 3270 twin pregnancies (2003 and 1209 conceived spontaneously and by IVF/ICSI, respectively) were identified. The proportion of twin pregnancies among all pregnancies was 3.4% (3332/97 278). Multiple regression analysis indicated that the incidences of gestational diabetes mellitus (adjusted odds ratio [AOR] = 1.42, 95% CI 1.10-1.83, p = 0.007), preterm premature rupture of membranes (AOR = 1.65, 95% CI 1.21-2.25, p = 0.002), placenta accreta spectrum (AOR = 2.12, 95% CI 1.42-3.17, p < 0.001) and postpartum hemorrhage (AOR = 1.38, 95% CI 1.02-1.86, p = 0.037) were significantly higher in the IVF/ICSI group than in the natural pregnancy group. Multivariate analysis also revealed that conception mode was not an independent risk factor for neonate outcomes. CONCLUSIONS: In twin pregnancies, IVF/ICSI is independently associated with multiple maternal complications, including gestational diabetes mellitus, preterm premature rupture of membranes and placenta accreta spectrum compared with spontaneous conception, although potential residual confounders due to indications for assisted reproductive technology exist.

Probe and Control of the Tiny Amounts of Dopants in BHJ Film Enable Higher Performance of Polymer Solar Cells.

To achieve efficient doping in polymer solar cells (PSCs), the dopant needs to be selectively located in the binary components of a bulk heterojunction (BHJ) film according to its polarity. The rarely studied n-type dopant is thoroughly examined in a simplified planar heterojunction (PHJ) device to address its favored location in the active layer. Results show that the n-dopant distribution in the acceptor layer or at the donor/acceptor interface produces enhanced device performance, whereas it harms the device when located in the donor layer. Based on the results, the benefit of n-type doping is then transferred to the highly efficient BHJ devices via a sequential coating procedure. The performance improvement is closely linked to the variations in the dopant's location in the BHJ film, which is carefully examined by the synchrotron techniques with delicate chemical sensitivity. More interestingly, the sequential coating procedure can be easily extended to the p-doped device only by changing the dopant's polarity in the middle layer. These findings pave the way for ambipolar doping in PSCs and enable performance improvement by molecular doping within the expectations.

Significance of Dopant/Component Miscibility to Efficient N-Doping in Polymer Solar Cells.

The uncertain dopant location in the bulk heterojunction (BHJ) film hinders the wide application of molecular doping in polymer solar cells (PSCs) as is in other organic devices. It is known that the interaction between the dopant and component governs the dopant distribution in the BHJ film and thus largely controls the effectiveness of molecular doping. After excluding the strong dopant/component interaction by forming the charge-transfer complex in the solution, we estimate the dopant/component miscibility by calculating the difference of Hansen's total solubility parameters (△δi-Hansen) and prove its correctness by contact angle measurements, and two model systems of poly[(2,6-(4,8-bis(5-(2-ethylhexyl)thiophe-2-yl)-benzo[1,2-b:4,5-b']dithiophene))-alt-(5,5-(1',3'-di-2-thienyl-5',7'-bis(2-ethylhexyl)benzo[1',2'-c:4',5'-c']dithiophene-4,8-dione))] (PBDB-T)/poly{[N,N'-bis(2-octyldodecyl)-naphthalene-1,4,5,8-bis(dicarboximide)-2,6-diyl]-alt-5,5'-(2,2'-bithiophene)} (N2200) and poly[4,8-bis(5-(2-ethylhexyl)-thiophene-2-yl)benzo[1,2-b;4,5-b']dithiophene-2,6-diyl-alt-(4-(2-ethylhexyl)-3-fluorothieno[3,4-b]thiophene-)-2-carboxylate-2-6-diyl]] (PCE10)/N2200 are selected to reveal the miscibility-photovoltaic performance relations. Only the material combination with large △δi-Hansen between the n-dopant (4-(1,3-dimethyl-2,3-dihydro-1H-benzoimidazol-2-yl)phenyl)dimethylamine (N-DMBI) and the donor polymer achieves enhanced photovoltaic performance. After that, we examine the doped morphology of polymer blends. Since the polymers' crystallizations are negatively affected by N-DMBI addition, we ensure the significance of n-doping on the enhanced device performance. Besides the dopant/polymer interaction, the solvent/polymer and solvent/dopant interactions are also considered to evaluate the kinetic effect on N-DMBI distribution by drawing the ternary phase diagram. We conclude that the kinetic morphological evolution does not change the miscibility-governed N-DMBI distribution in the BHJ film. Finally, we provide a direct relationship between the N-DMBI position and the device property by fabricating the bilayer devices. The enhancement of photovoltaic performances is observed in both material systems only if the N-DMBI distributes in N2200. Our work outlines a basis for using the dopant/component interaction and ternary phase diagram to predict the dopant distribution before extensive experiments. It significantly reduces the trial-and-error work and increases the reliability of molecularly doped PSCs.

Organic photovoltaic cell with 17% efficiency and superior processability.

The development of organic photoactive materials, especially the newly emerging non-fullerene electron acceptors (NFAs), has enabled rapid progress in organic photovoltaic (OPV) cells in recent years. Although the power conversion efficiencies (PCEs) of the top-performance OPV cells have surpassed 16%, the devices are usually fabricated via a spin-coating method and are not suitable for large-area production. Here, we demonstrate that the fine-modification of the flexible side chains of NFAs can yield 17% PCE for OPV cells. More crucially, as the optimal NFA has a suitable solubility and thus a desirable morphology, the high efficiencies of spin-coated devices can be maintained when using scalable blade-coating processing technology. Our results suggest that optimization of the chemical structures of the OPV materials can improve device performance. This has great significance in larger-area production technologies that provide important scientific insights for the commercialization of OPV cells.

Achieving High Doping Concentration by Dopant Vapor Deposition in Organic Solar Cells.

Compared with the interfacial doping, molecular doping in bulk heterojunction (BHJ) is a more direct but challenging approach to optimize the photovoltaic performance in organic solar cells (OSCs). One of the main obstacles for its success is the low doping concentration because of the morphological damage. Starting from the phase diagram analysis, we discover that the unpreferred good miscibility between the p-type dopant and the acceptor leads to incorrect dopant dispersion, which reduces the achievable doping content. To overcome this, we use sequential doping by vapor annealing instead of blend solution doping, and we achieve the high doping concentration without sacrificing the blend film morphology. Benefiting from the undamaged film, we fulfill improved photovoltaic performance. Our positive results reveal the feasibility of high-level doping in complex organic BHJ films. It is believed that doping at high concentration potentially enlarges the extent of tunable range on electronic properties in OSCs and indicates greater improvement for device performance.

Balancing Crystal Size in Small-Molecule Nonfullerene Solar Cells through Fine-Tuning the Film-Forming Kinetics to Fabricate Interpenetrating Network.

The nanoscale interpenetrating network of active layer plays a key role in determining the exciton dissociation and charge transport in all small-molecule nonfullerene solar cells (AS-NFSCs). However, fabricating interpenetrating networks in all small-molecule blends remains a critical hurdle due to the uncontrolled crystallization behavior of small molecules. In this study, we proposed that the balanced crystal size between the donor and the acceptor is an essential prerequisite to construct optimal interpenetrating networks. We also provided a solvent additive strategy to reduce the gap of crystal size between the donor and the acceptor in S-TR:ITIC all small-molecule blend system through manipulating the solution state and film-forming kinetics. As a result, the crystal size of S-TR decreased and the crystal size of ITIC increased, leading to nanoscale interpenetrating networks. This optimized morphology improved the exciton dissociation efficiency and suppressed the bimolecular recombination, achieving almost double power conversion efficiency compared to the reference device. This work demonstrates that manipulation of the balanced crystal size of donor and acceptor may be a key to further boost the efficiency of AS-NFSCs.

mAChRs activation induces epithelial-mesenchymal transition on lung epithelial cells.

BACKGROUND: Epithelial-mesenchymal transition (EMT) has been proposed as a mechanism in the progression of airway diseases and cancer. Here, we explored the role of acetylcholine (ACh) and the pathway involved in the process of EMT, as well as the effects of mAChRs antagonist. METHODS: Human lung epithelial cells were stimulated with carbachol, an analogue of ACh, and epithelial and mesenchymal marker proteins were evaluated using western blot and immunofluorescence analyses. RESULTS: Decreased E-cadherin expression and increased vimentin and α-SMA expression induced by TGF-ß1 in alveolar epithelial cell (A549) were significantly abrogated by the non-selective mAChR antagonist atropine and enhanced by the acetylcholinesterase inhibitor physostigmine. An EMT event also occurred in response to physostigmine alone. Furthermore, ChAT express and ACh release by A549 cells were enhanced by TGF-ß1. Interestingly, ACh analogue carbachol also induced EMT in A549 cells as well as in bronchial epithelial cells (16HBE) in a time- and concentration-dependent manner, the induction of carbachol was abrogated by selective antagonist of M1 (pirenzepine) and M3 (4-DAMP) mAChRs, but not by M2 (methoctramine) antagonist. Moreover, carbachol induced TGF-ß1 production from A549 cells concomitantly with the EMT process. Carbachol-induced EMT occurred through phosphorylation of Smad2/3 and ERK, which was inhibited by pirenzepine and 4-DAMP. CONCLUSIONS: Our findings for the first time indicated that mAChR activation, perhaps via M1 and M3 mAChR, induced lung epithelial cells to undergo EMT and provided insights into novel therapeutic strategies for airway diseases in which lung remodeling occurs.

A novel mutant allele specific amplification and electrochemiluminescence method for the detection of point mutation in clinical samples.

A novel mutant allele specific amplification (MASA) and electrochemiluminescence (ECL) method for point mutation detection is proposed. Briefly, the target gene was amplified by a biotinylated mutant specific sense primer and a Ru(bpy)(3)(2+) (TBR)-labeled universal antisense primer. Only the mutant allele can be selectively amplified by the mutant specific primer pair. Then, the MASA product was captured onto the streptavidinylated magnetic beads through biotin-streptavidin linkage and detected by measuring the ECL emission of TBR. The method was applied to detect a possible point mutation at codon 12 of K-ras oncogene in 30 colorectal cancer (CAC) clinical samples. The experimental results show that the method can detect K-ras mutant in a 5000-fold excess of wild-type allele. Furthermore, different kinds of mutations can be clearly discriminated. The point mutation was found in 15 (50%) out of 30 CAC samples. This novel MASA-ECL method could potentially become a sensitive, specific, simple, rapid and safe approach for point mutation detection.

Development and application of a capillary electrophoresis-electrochemiluminescent method for the analysis of enrofloxacin and its metabolite ciprofloxacin in milk.

Enrofloxacin (ENR) is a fluoroquinolone developed exclusively for the use in veterinary practice for the treatment of respiratory and gastrointestinal infections, and ciprofloxacin (CIP) is its main active metabolite. Their contents are regulated by the EU Council Regulation no. 2377/90 in animal edible tissues. We developed a sensitive and rapid method for the determination of ENR and CIP by capillary electrophoresis (CE) with electrochemiluminescence (ECL) detection. The method is based on the detection of aliphatic tertiary or secondary amino moieties in ENR and CIP with end-column tris(2,2-bipyridyl)ruthenium(II) electrochemiluminescence. Parameters that affect separation and detection were optimized. Under the optimized conditions, the calibration functions were linear in the range of 0.03-1 microg ml(-1) for ENR and 0.05-1.2 microg ml(-1) for CIP. The detection limits of ENR and CIR were 10 ng ml(-1) and 15 ng ml(-1), respectively, based on the signal-to-noise ratio of 3. The relative standard derivations of the peak height and the migration time for ENR and CIP were less than 4.13%. The developed method was successfully applied to determine ENR and CIP in milk with a solid-phase extraction procedure.

PCR-free quantitative detection of genetically modified organism from raw materials. An electrochemiluminescence-based bio bar code method.

A bio bar code assay based on oligonucleotide-modified gold nanoparticles (Au-NPs) provides a PCR-free method for quantitative detection of nucleic acid targets. However, the current bio bar code assay requires lengthy experimental procedures including the preparation and release of bar code DNA probes from the target-nanoparticle complex and immobilization and hybridization of the probes for quantification. Herein, we report a novel PCR-free electrochemiluminescence (ECL)-based bio bar code assay for the quantitative detection of genetically modified organism (GMO) from raw materials. It consists of tris-(2,2'-bipyridyl) ruthenium (TBR)-labeled bar code DNA, nucleic acid hybridization using Au-NPs and biotin-labeled probes, and selective capture of the hybridization complex by streptavidin-coated paramagnetic beads. The detection of target DNA is realized by direct measurement of ECL emission of TBR. It can quantitatively detect target nucleic acids with high speed and sensitivity. This method can be used to quantitatively detect GMO fragments from real GMO products.

An improved electrochemiluminescence polymerase chain reaction method for highly sensitive detection of plant viruses.

Recently, we have reported an electrochemiluminescence polymerase chain reaction (ECL-PCR) method for detection of genetically modified organisms. The ECL-PCR method was further improved in the current study by introducing a multi-purpose nucleic acid sequence that was specific to the tris(bipyridine) ruthenium (TBR) labeled probe, into the 5' terminal of the primers. The method was applied to detect plant viruses. Conserved sequence of the plant viruses was amplified by PCR. The product was hybridized with a biotin labeled probe and a TBR labeled probe. The hybridization product was separated by streptavidin-coated magnetic beads, and detected by measuring the ECL signals of the TBR labeled. Under the optimized conditions, the experiment results show that the detection limit is 50 fmol of PCR products, and the signal-to-noise ratio is in excess of 14.6. The method was used to detect banana streak virus, banana bunchy top virus, and papaya leaf curl virus. The experiment results show that this method could reliably identity viruses infected plant samples. The improved ECL-PCR approach has higher sensitivity and lower cost than previous approach. It can effectively detect the plant viruses with simplicity, stability, and high sensitivity.